PMID- 16035311 TI - [Drug-induced diabetes mellitus: the exemple of atypical antipsychotics]. AB - As an example of iatrogenic diabetes, we discuss the problem of diabetes mellitus in patients receiving atypical antipsychotics. The risk of diabetes mellitus appears to be different according to various molecules with, by decreasing order, clozapine, olanzapine, risperidone and quetiapine, and finally amisulpride, aripiprazole and ziprasidone. A careful analysis of published case reports and series indicates the existence of two different problems: 1) the rather common development of impaired glucose tolerance or diabetes mellitus (often associated with metabolic syndrome) related to weight gain in individuals at risk for type 2 diabetes; and 2) the occurrence of rare cases of acute metabolic episodes with severe ketoacidosis and/or pancreatitis whose pathophysiological mechanisms remain largely unknown. Generally speaking, the pathophysiology involves both increased insulin resistance and deficient insulin secretion. Cautious metabolic monitoring of patients receiving atypical antipsychotics is recommended, and the selection of the appropriate drug should be influenced by the metabolic profile of the various molecules and the metabolic risk of the patients who should be treated with atypical antipsychotics. PMID- 16035312 TI - [Risk of hypoglycaemia: therapeutical implications]. AB - Hypoglycaemia is defined as a plasma glucose concentration equal or below 54 mg/dl (3 mmol/l). It represents the most frequent acute metabolic complication among diabetic patients. Rarely fatal in patients with type 1 diabetes, it markedly alters the quality of life and hinders the achievement of a good metabolic control. Hypoglycaemia in type 2 diabetic patients treated with insulin secreting agents is less frequent, but it may be more dangerous in case of prolonged coma. Numerous intrinsic and extrinsic factors may influence the kinetics of plasma glucose concentrations among diabetic patients, which limits the prevision, and thus the prevention, of hypoglycaemic episodes. However, some basic rules, repeatedly emphasized during the multidisciplinary educational process, allow minimizing both the number and the severity of hypoglycaemic episodes. The present paper briefly describes some clinical situations increasing the risk of hypoglycaemia in diabetic patients treated with insulin or insulin secreting agents, emphasizing upon important therapeutic implications. PMID- 16035313 TI - [Metabolic emergencies related to diabetes mellitus: ketoacidosis and hyperosmolar state]. AB - Diabetic ketoacidosis and hyperglycaemic hyperosmolar state are relatively frequent metabolic emergencies. Such entities complicate type 1 and type 2 diabetes mellitus, respectively. Diagnosis is ultimately provided by blood and urine analysis. The pathophysiology is mainly based on insulin privation regarding ketoacidosis and resistance to insulin in hyperglycaemic crisis, with an additional deleterious role of counterregulatory hormones. Prognosis of such complications remains generally severe. Management is based on insulin infusion, fluid resuscitation as well as concomitant compensation for electrolytes losses. PMID- 16035314 TI - [A central pathological mechanism explaining diabetic complications?]. AB - Diabetes mellitus is associated to micro- and macro-vascular lesions responsible for myocardial infarction, nephropathy, retinopathy and polyneuropathy. Four main pathogenic mechanisms have been proposed, all associated with hyperglycaemia: 1) increased flux in the polyol pathway; 2) increased flux in the hexosamine pathway; 3) protein kinase C activation; and 4) increased formation of advanced glycation endproducts. A common mechanism seems to play a central role in the activation of these various pathways. Indeed, an increased production of free radicals by mitochondria induced by hyperglycaemia may be responsible for the observed metabolic disturbances. The present article describes that theory and presents its possible therapeutic implications. PMID- 16035315 TI - [Diabetic retinopathy]. AB - Diabetic retinopathy remains a serious cause of blindness in the Western World. Its major clinical signs and management are reviewed, with particular focus on advances in the diagnosis and treatment of diabetic macular edema. PMID- 16035316 TI - [Diabetic peripheral neuropathy]. AB - Peripheral neuropathy is a very frequent consequence of diabetes mellitus. Its clinical expression is quite variable. A specific therapy is sometimes necessary. Early diagnosis of diabetic neuropathy is a cornerstone of patient's follow-up. Differential diagnosis of diabetic neuropathy is sometimes difficult from another type of neuropathy or a focal, even systemic, disease. It is mandatory to know how a diabetic neuropathy may express. Pathophysiological mechanisms involved in diabetic neuropathy are complex and interrelated. Hyperglycaemia alone, even mild or moderate, vascular disorders and dysimmune factors may be combined to induce axonal injury. Glycaemic control is the cornerstone of effective treatment for neuropathy associated with diabetes. Specific pain control and therapies of autonomic disturbances are regularly required. PMID- 16035317 TI - [Diabetic cardiac autonomic neuropathy]. AB - Cardiac autonomic neuropathy (CAN) is a common complication of diabetes mellitus, which is associated with a higher risk of morbidity and mortality. It can be detected by analyzing spontaneous (Holter) or provoked (Ewing's test battery) changes in heart rate and arterial blood pressure. Baroreflex gain is a specific index of great interest. Our laboratory has acquired a large experience in the assessment of CAN in diabetic patients. We use the Finapres, a device that allows continuous noninvasive monitoring of blood pressure and heart rate, and a special and discriminative active orthostatic manoeuvre, the "squatting" test (standing squatting-standing). PMID- 16035318 TI - [Erectile dysfunction]. AB - Diabetes is a frequent aetiology of erectile dysfunction. The prevalence of erectile dysfunction in diabetics is three times higher than in the general population. Erectile dysfunction in diabetics is multifactorial. Phosphodiesterase type 5 inhibitors represent the treatment of first intention. PMID- 16035319 TI - [Diabetic gastroparesis]. AB - Diabetic gastroparesis corresponds to symptomatic as well as asymptomatic gastric retention without organic abnormality of stomach, pylorus or gut. This complication associated with autonomic neuropathy is found in about 50% of patients with type 1 and type 2 diabetes. It may be clinically important when it is associated with gastrointestinal symptoms limiting quality of life, alterations in glycaemic control and changes in oral drug absorption. In addition, acute changes in blood glucose concentration affect gastric motor function: gastric emptying is slowed down during hyperglycaemia and accelerated during hypoglycaemia. The diagnosis of gastroparesis may be confirmed by scintigraphy assessment of gastric emptying, preferably using a solid meal. Unfortunately, treatment options remain limited and often unsatisfactory. They first rely on life-style and dietary modifications. If necessary, pharmacological agents (metoclopramide, domperidone, cisapride, and erythromycin) may be considered. Cisapride is actually the most powerful agent for chronic use, but the risk of cardiac toxicity (increase of QT with "torsade de pointe") limits its general use. In some diabetic patients, gastroparesis may contribute to erratic glucose excursions, with precocious postprandial hypoglycaemia, late hyperglycaemia, and/or delayed recovery from hypoglycaemia after carbohydrate ingestion. Sometimes, the initiation of intensive insulin therapy and the use of prokinetic drugs could lead to significant improvement of blood glucose control in patients with diabetic gastroparesis. PMID- 16035320 TI - [The diabetic foot]. AB - The pathophysiological mechanisms underlying a diabetic foot disease are complex and multifactorial, including neuropathy, ischemia, infection and abnormal foot biomechanics. All these factors are often intricated and source of delayed wound healing. Insight in the pathophysiology of the diabetic foot provides a comprehensive basis for a protocol of primary and secondary preventive care. Since non-enzymatic glycosilation of proteins and of connective tissue underlies structural changes in vessels, nerves and osteo-articular structures, a rigid control of blood glucose levels is of paramount importance. Early recognition of the etiology of foot lesions and prompt management of foot ulcers are essential for successful outcome. Aggressive treatment of infections, clinical assessment and correction of vascular occlusive disease (diabetic macroangiopathy), adequate wound care and appropriate off-loading (pressure relief) of the ulcer are essential steps in the treatment protocol. It is not surprising that optimal management of the diabetic foot requires a multidisciplinary approach in a Diabetic Foot Clinic, coordinating care-provisions by a team of diabetologist, infectiologist, vascular surgeon, interventional radiologist, plastic surgeon, podiatrist and specialized nurse. Applying evidence-based multidisciplinary treatment results in a 50% reduction of major lower-limb amputation in this high risk group. PMID- 16035321 TI - [Coronary artery disease associated to diabetes mellitus]. AB - Diabetes mellitus represents a major coronary risk factor. Diabetes promotes atherosclerotic plaque development through different mechanisms among which oxidative stress induced by hyperglycaemia appears to play a major role. Knowledge of pathophysiologic mechanisms associated with diabetes helps to better understand the rationale for treatment. Besides a tight control of all cardiovascular risk factors, prescription of drugs aiming of stabilization or even reduction of atherosclerosis should be systematically recommended. These drugs include statines, ACE inhibitors and aspirin eventually combined with clopidogrel. The use of beta blockers should be encouraged in case of stable angina. Finally, any patient with symptomatic modification suggesting acute coronary syndrome should be considered for myocardial revascularization. PMID- 16035322 TI - [Silent coronary artery disease in diabetic patients. New guidelines]. AB - More than half of diabetic individuals will die from a coronary event. Coronary artery disease often presents an atypical form among diabetic subjects. Silent myocardial ischaemia may be detected in 20 to 35% of diabetic patients with associated cardiovascular risk factors. When a coronarography is performed in patients with silent myocardial ischaemia, it demonstrates significant coronary stenosis in one to two thirds of patients. The prognosis of diabetic patients with silent myocardial ischaemia is associated with a higher incidence of cardiac events in the next three years, especially when silent ischaemia is associated with angiographically coronary stenosis. French guidelines jointly published in 2004 by the ALFEDIAM and the French Society of Cardiology propose the search for silent myocardial ischaemia--in diabetic patients with peripheral arteriopathy or overt nephropathy with proteinuria,--in diabetic patients with microalbuminuria and two other classical cardiovascular risk factors,--in a sedentary diabetic patient who wants to begin a physical activity,--in type I diabetic patients above 45 years or with a disease lasting for more than 15 years and in type 2 diabetic patients above 60 years or with a known disease lasting for more than 10 years, when at least two other traditional cardiovascular risk factors are present. Besides the standard annual electrocardiogram, these high risk patients should benefit first from an exercise test or when the latter is impossible, under-maximal or doubtful, from a myocardial scintigraphy combined with dipyridamole injection or from a stress echocardiography. The demonstration of a silent myocardial ischaemia should lead to a coronarography when the general status of the patient and the absence of severe comorbidities allow considering a coronary revascularisation procedure in these diabetic patients. PMID- 16035323 TI - [Congestive heart failure and diabetes mellitus]. AB - Diabetic patients frequently develop heart failure and have a guarded prognosis because of the combination of diabetic cardiomyopathy, hypertension and ischaemic heart disease. The lack of patient awareness of the association between diabetes and cardiovascular disease contributes to the risk of heart failure in the diabetic population. Therefore, physicians should consider the risk of comorbidities and use appropriate screening tests to achieve early identification and initiate preventive strategies. There is evidence suggesting that adequate glycaemic control may improve cardiac metabolism and myocardial function in diabetics with heart failure. Improvement in cardiac function engendered by neurohumoral inhibition is associated with a decrease in mortality that is at least as great in the diabetic patients with heart failure as it is in the non diabetic patients. However, several therapeutic interventions have not yet been specifically validated in diabetic subjects. Thus, physicians should be encouraged to use therapies tested in the diabetic population, such as B-blockers and ACE inhibitors. Diabetic patients would benefit from more aggressive preventive programs aimed at reducing the incidence of cardiovascular morbidity and mortality. PMID- 16035324 TI - [Alterations in natural immunity and risk of infection in patients with diabetes mellitus]. AB - This review addresses the recent litterature devoted to the risk of severe infections in patients with diabetes and to the potential influence of diabetes on the function of natural immunity. Although much controversy still exists regarding the incidence of infections in diabetic patients, several studies confirm that diabetes mellitus is associated with an increased severity and mortality in community acquired pneumonia. Furthermore, the risk of severe bacteremia (especially associated with Streptococcus pneumoniae) is higher in diabetic patients. Polynuclear neutrophils are clearly influenced by the diabetic state. On the one hand, their antimicrobial function is inhibited by hyperglycaemia, due to inhibition of G6PD or diversion of NADPH in the polyol pathway; on the other hand, the AGE/RAGE/NF-kappaB pathway involved in the pathogenesis of chronic complications of diabetes could also amplify inflammatory systemic manifestations associated with infections and play a role in the higher mortality rate observed in diabetic subjects with severe infections. These observations argue for the systematic vaccination of all diabetic patients against influenza and Streptococcus pneumoniae, for the reappraisal of diabetes as a significant pejorative risk factor in community acquired pneumonia and for intensive insulin therapy in all diabetic patients with severe infection. PMID- 16035325 TI - [Mucormycosis and diabetes: three cases reported]. AB - Mucormycosis is a rare and often fatal fungal infection. It mainly occurs when predisposing factors such as diabetic ketoacidosis are present. We report three cases of rhino-orbito-cerebral, rhino-orbital and pulmonary mucormycosis in patients with diabetic ketoacidosis. The diagnosis was reached after identification of hypae of the order mucorales using pathology and mycology examinations. A fatal issue occured in two cases and one case survived after adequate therapy. PMID- 16035326 TI - [Asymptomatic bacteriuria: which management in patients with diabetes mellitus?]. AB - Most of the research about asymptomatic bacteriuria (ASB) in patients with diabetes mellitus has been performed in female patients, with a prevalence of approximately 7-13 %, about 3 to 4 times higher than that reported for nondiabetic women. Poor metabolic control, as assessed by haemoglobin A1c or glucosuria, is not associated with increased ASB. The latter is associated with an increased risk of symptomatic urinary tract infection among patients with type 2 diabetes, but not type 1 diabetes. These infections tend to be more complicated and caused by more resistant organisms than reported for nondiabetic patients. Despite this, systematic screening for and treating ASB have not been shown to reduce long-term complications, such as accelerated progression to arterial hypertension or renal failure, or symptomatic urinary tract infections (including pyelonephritis) or hospitalization for these infections. Thus, available evidence does not support systematic screening and antimicrobial treatment of ASB among patients with diabetes mellitus. PMID- 16035327 TI - [Cutaneous stigmata of diabetes mellitus]. AB - A number of relationships exist between diabetes mellitus and a series of cuteanous disorders. Skin lesions may reveal diabetes in a more or less specific way. They may also represent complications supervening in an already treated diabetic patient. Some of these dermatoses (acanthosis nigricans, purpuric and pigmented capillaritis) are markers of macrovascular complications. The same disorders and some others (xerosis, Dupuytren's disease) are more frequently associated with microangiopathy in Type II diabetes. Other skin diseases (alopecia areata, vitiligo) are markers of auto-immunity in Type I diabetes. PMID- 16035328 TI - [The so-called dry skin of the diabetic patient]. AB - Rough skin or xerosis, known by the laity as dry skin, results from a defect in the stratum corneum desquamation which normally remains imperceptible. This condition is negatively influenced by winter climatic conditions. Atopic dermatitis and trophic changes related to post-menopause, ageing, diabetes mellitus, as well as other endrocrine and metabolic disturbances are also involved in this process. Xerosis can be improved at various degrees by emollients, humectants, hydrating agents and squamolytic agents. PMID- 16035329 TI - [Cutaneous complications of insulin therapy. A drug-induced condition on the decline]. AB - Insulin therapy is increasingly used. Hopefully, its complications are steadily decreasing in incidence since the introduction of new generations of insulin. The main cutaneous side effects were varied, including lipoatrophy, lipohypertrophy, allergic reactions, infectious abcesses and some idiosyncratic reactions. PMID- 16035330 TI - [Type 2 diabetes, in the heart of the metabolic syndrome: plea for a global approach]. AB - Type 2 diabetes is frequently associated with other comorbidities among which abdominal obesity, arterial hypertension, dyslipidaemia and a prothrombotic state. All these cardiovascular risk factors belong to the so-called metabolic syndrome. The appropriate management of a patient with type 2 diabetes requires a global approach targeting each risk factor in order to reduce cardiovascular morbidity and mortality. Microalbuminuria is correlated with markers of the metabolic syndrome in diabetic patients as shown by the DANA survey performed among general practitioners from the areas of Liege and Antwerp and, in this type of patients, is considered as a prognostic factor of cardiovascular complications. The OCAPI study, performed among Belgian internists and diabetologists, demonstrated that two thirds of diabetic patients have a metabolic syndrome and that the objectives are far from being reached in clinical practice, especially as far as lipid and arterial pressure control is concerned. Interestingly, the Danish STENO-2 study demonstrated that an intensive multi-risk approach reduces the incidence of both micro- and macroangiopathic complications by more than 50%. This challenge represents a major public health issue considering the high and increasing prevalence of type 2 diabetes and the major burden associated to this disease. PMID- 16035331 TI - [Dyslipidaemia and diabetes mellitus]. AB - Cardiovascular diseases are the main complications in diabetes. The lipid pattern includes high triglycerides, low HDL and increased small dense LDL. The treatment strategy is based upon the European Guidelines which insist on global risk evaluation and recommend more severe lipid targets for diabetic than for non diabetic patients. After changes of lifestyle and control of risk factors, statins or fibrates are the drugs of choice, even if fibrates have specific impact on patients with diabetes or the metabolic syndrome. PMID- 16035332 TI - [Menopause-related risks and hormone therapy in diabetic women]. AB - Prevalence of diabetes mellitus (types 1 and 2) in postmenopausal women is about 10-20% according to age. It can be associated with a metabolic syndrome in about 60% of cases, thereby severely increasing cardiovascular risk (among others) in these women. Estrogen or estrogen-progestin replacement therapy does not usually impair diabetes control. It will be submitted to the same indications/contraindications, in relation with the risk/benefit balance, as for all other postmenopausal women. However, increased risks inherent to diabetic subjects concerning metabolism, coagulation/hemostasis, and cardiovascular disease, should be considered. Therefore, estrogen in minimal effective dosages, eventually by transdermal route, as well as metabolically neutral progestins near to progesterone should be preferred. In case of pre-existing or occurring vascular problems, a careful approach or even suppression of replacement therapy should prevail. PMID- 16035333 TI - [Role of clinical chemistry in the assessment and follow up of a diabetic patient]. AB - Clinical chemistry is essential in the management of a patient with diabetes mellitus. It plays a major role in the diagnosis by measuring plasma glucose levels, in the fasting state or two hours after an oral glucose load. It helps in the definition of the type of diabetes by measuring plasma C-peptide concentration, a marker of residual endogenous insulin secretion, or by detecting islet cell antibodies, a marker of autoimmune disease. Finally, clinical chemistry is essential in assessing the quality of overall blood glucose control thanks to the dosage of glycated haemoglobin (HbA1c) and in demonstrating the adequate control of cardiovascular risk factors associated to the metabolic syndrome, especially dyslipidaemia. PMID- 16035334 TI - [Hospital in-patients with diabetes]. AB - As the diabetic population is becoming older, hospitalization is more frequent and of longer duration with increased costs. Hospital stay could be a good opportunity to improve diabetes care. All observational studies demonstrate that improving metabolic control reduces mortality, morbidity and hospitalisation costs. For critically ill patients and those referred for myocardial infarction or submitted to cardiac surgery, randomised controlled studies have proven the efficacy of strict metabolic control and defined the targets to be reached. Unfortunately, for many reasons, it is difficult to treat the patients to the targets and in non specialized departments, diabetes control may even be worse during hospitalisation. We urgently need to find innovating solutions to mitigate the consequences of the reduced number of competent nurses and doctors in the field of diabetes. PMID- 16035335 TI - [Therapeutic education of the diabetic patient]. AB - Therapeutic education is a patient centred approach focused on patients' needs, values and strategies. It allows not only to increase patients' knowledge and skills on the disease, but also on their treatments. It brings a better quality of life, an increased therapeutic compliance, and decreases complications. The most difficult part of therapeutic education occurs when patients must change their behaviour. Motivational interviewing techniques are of great support and allow to prepare and support patients in "step by step" progressive changes. It is essential to work on resistance to change. The ambivalence in the choice of a new lifestyle must be measured, discussed and negotiate. Patients become partners and health care providers become coaches. To negotiate objectives must allow patients to choose their own strategies which should cost them minimum psychological efforts and bring them maximum benefits. The efficacy of therapeutic education has not to be proved any more: 80% less of amputations at 10 years in diabetic patients, 50% of stable body weight after weight loss at 5 years, etc. In conclusion, therapeutic education is part of a humanist medicine centred on patients which allow them to take care of their own treatment, in order to improve their quality of life, therapeutic compliance and potential complications. The health care providers teach, train, negotiate, motivate and accompany their patients in the long-term follow-up of their diseases. PMID- 16035336 TI - [Multidisciplinary educational health programme for type 2 diabetic patients in primary care]. AB - Type 2 diabetic patients followed in primary care rarely benefit from a structured educational approach in spite of the fact that the intervention of a multidisciplinary team has proven its efficacy for improving metabolic control. We describe a pilot project, which aimed at implementing an ambulatory multidisciplinary programme and at evaluating the feasibility of an educational module in type 2 diabetic patients followed in primary care. Four general practitioners (GP) selected 25 type 2 diabetic patients, not well controlled or with poor knowledge of their disease. Fourteen health care providers developed an intervention module comprising, over 4 months, an initial interview with the GP, three interviews with a dietician, two with a nurse and three with a psychologist. Besides studying the feasibility of the project, the primary objective was to analyze the subjective feeling of both patients and heath care providers participating to the project. A secondary objective was to compare some parameters of control (HbA1c, body weight) before and after the intervention in these 25 patients, and in 10 control patients followed as usual in general practice. The multidisciplinary educational intervention was well appreciated by diabetic patients whose adhesion was excellent. Health care provider's mobilization was enthusiastic, and all emphasized the human positive aspects of such a multidisciplinary approach. Preliminary analysis demonstrates beneficial effects of the educational programme. PMID- 16035337 TI - [The role of diabetes associations]. AB - The first diabetes association was created in 1926, soon after the first clinical use of insulin, in Portugal. In Belgium, the "Association Belge du Diabete" was founded in 1942, and the International Diabetes Federation (IDF) was created in 1950 with at that time 16 associations, in 2005 more than 180 associations. The aims of these associations are: --Defence of the rights of the diabetic people in all fields, --Education of the patients and their family as well as of the healthcare professionals on diabetes, --Representation of people with diabetes and of healthcare professionals involved in diabetes at all levels, keeping in mind the major importance of solidarity, --Support to the clinical as well as the fundamental research, --Increase of the global knowledge concerning diabetes, - Actions to favour positive changes, --Act as the global advocate of the cause of diabetes --Facilitate the access to medication (especially insulin), which still is a major problem in many parts of the world, Knowing the explosion of diabetes throughout the world with all the medical, economical, professional, social and human problems related to this major increase, it is obvious that the role of diabetes associations still is and will be even more and more important. PMID- 16035338 TI - [Social aspects of diabetes mellitus]. AB - A diabetic patient may suffer from social discriminations. We will briefly describe the problems related to licence driving, employment and insurances. Diabetic associations work to attenuate such discriminations and try to provide a practical help to diabetic patients confronted to such difficulties. PMID- 16035339 TI - [The role of diabetes convention centres in Belgium]. AB - We briefly present the modes of functioning of Diabetes Convention Centres in Belgium. In those hospital centres, patients with both type 1 or type 2 diabetes, treated by at least two insulin injections per day, benefit of an intensive educational programme by a multidisciplinary team and receive free of charge material for home blood glucose monitoring, in order to optimize diabetes management. The collaboration between convention centres and general practitioners should be reinforced (share-care), especially to improve the management of type 2 diabetic patients, who are increasingly treated with various insulin regimens. PMID- 16035340 TI - [Initiave to promote quality of care and epidemiology of diabetes mellitus]. AB - Since 2001, the "Diabetes Convention Centres" in Belgium participated to a survey, called IPQED ("Initiative for the Promotion of Quality and Epidemiology of Diabetes mellitus), every 12-18 months. This project aims at systematically and anonymously collecting a limited but significant set of data (anamnesis, clinical examination, clinical chemistry: according to DiabCare) in a large sample (about 10%) of type 1 or type 2 diabetic patients followed in the Diabetes Convention Centres (inclusion criteria: at least two injections of insulin per day). IPQED has three main objectives: 1) to optimise the quality of care using a benchmarking process and the creation of quality circles; 2) to provide to health authorities some general advise concerning care for diabetic patients, in general, and in the Convention Diabetes Centres, in particular; and 3) to collect Belgian data among a large cohort of diabetic patients (around 8.000 per survey) that could be used for epidemiological studies. PMID- 16035341 TI - [Diabetes mellitus: epilogue]. PMID- 16035342 TI - Inhibition of return and repetition priming effects in localization and discrimination tasks. AB - Inhibition of return (IOR) refers to slower responding to a stimulus that is presented at the same, rather than a different location as a preceding, spatially nonpredictive, stimulus. Repetition priming refers to speeded responding to a stimulus that duplicates the visual characteristics of a stimulus that precedes it. IOR and repetition priming effects interact in nonspatial discrimination tasks but not in localization tasks; three experiments examined whether this is due to processing differences or due to response differences between tasks. Two stimuli, S1 and S2, occurred on each trial. In Experiment 1, S1 and S2 were both peripheral arrows; in Experiment 2, S1 was a central arrow and S2 was a peripheral nondirectional rectangle; in Experiment 3, S1 was a peripheral nondirectional rectangle and S2 was a peripheral arrow. S1 never required a response; S2 required a localization or a discrimination response. Despite evidence that form information was likely extracted from the arrow stimuli, the localization task revealed no repetition priming: IOR occurred regardless of shared visual identity of the S1 and S2 arrows. The discrimination task revealed IOR only when the visual identity changed from S1 to S2; otherwise, facilitation occurred. These results suggest that IOR is masked by repetition priming only when the response depends on the explicit processing of form information; repetition priming does not occur when such information is extracted automatically but is task (and response) irrelevant. PMID- 16035343 TI - Examining task difficulty and the time course of inhibition of return: detecting perceptually degraded targets. AB - The ability to efficiently direct visual attention to salient features in the environment is a critical function of the visual system. The finding that people are slower to detect a target that appears at a previously cued location is thought to reflect a mechanism known as inhibition of return (IOR). Past research has shown that difficult target discriminations result in a greater amount of time needed to inhibit previously attended locations (i.e., a delayed onset of inhibition), suggesting that task difficulty plays a critical role in the allocation of attention. In this study, IOR was measured at a wide range of SOAs while participants detected either a perceptually degraded target or a standard, high luminance target. When responses were made to a perceptually degraded target, the time course of IOR was delayed by approximately 250 ms (relative to the control group), suggesting that the difficulty in detecting targets also influences the allocation of attention. The results are consistent with the notion that IOR is not simply a reflexive subcortical mechanism but rather involves top-down attentional control settings. PMID- 16035344 TI - Spatial attention as a necessary preliminary to early processes in reading. AB - One major theory of the relation between spatial attention and visual word recognition holds that the former is a necessary condition for the latter to begin. A different major theory asserts that although spatial attention can facilitate the latter, it is not a necessary condition. These two theories were pitted against each other experimentally. Spatial attention was operationalized in terms of the effect of a spatial precue on the time to name a target word that appeared above or below fixation. A masked prime word was presented before the target. The critical difference between experiments was cue validity (50% in Experiments 1a and 2a and 100% in Experiments 1b and 2b). Repetition priming was observed when the prime appeared in the uncued prime location in Experiments la and 2a but not in Experiments 1b or 2b. These results are inconsistent with the claim that visual word recognition does not depend on spatial attention. Discussion centres on the distribution of spatial attention across target locations as a function of cue validity. PMID- 16035345 TI - The time course of the contingent spatial blink. AB - Attentional capture is the unintentional deployment of attention to a task irrelevant but attentionally salient object. The contingent involuntary orienting hypothesis states that it occurs only if a distractor's property matches current top-down attentional control settings (Folk, Remington, & Johnston, 1992). Folk, Leber, and Egeth (2002) found that monitoring a central RSVP stream for a coloured target led to spatial attentional capture by a peripheral distractor that matched the target colour. Using a similar paradigm, we explored the time course of this spatial blink. Implications of this study for current accounts of the attentional capture phenomenon are discussed. PMID- 16035346 TI - Dissociating stimulus-stimulus and response-response effects in the Stroop task. AB - The separate semantic and response competition interactions between colour and word processing in a manual Stroop task were evaluated by comparing three trial types. Identity trials are both semantically compatible and response compatible (e.g., BLUE in the colour blue), different response trials are both semantically incompatible and response incompatible (e.g., BLUE in the colour green, where blue and green have different response keys), and same response trials are semantically incompatible and response compatible (e.g., the word BLUE in the colour red, where blue and red have the same key press response). Ink colours were embedded in two different word types, colour words, and colour associates. The results using colour words replicated the findings of De Houwer (2003) and demonstrated both a semantic effect (a difference between same response trials and identity trials) and response competition (a difference between same response trials and different response trials). In contrast, the results using colour associates provided evidence for only a semantic effect. These findings support interpretations of the colour associate Stroop effect that attribute the effect to semantics, but challenge Klein's (1964) response competition account and Sharma and McKenna's (1998) claim that the effect of colour associates is dependent on verbal responding. The results confirm that the Stroop colour-word task appears to involve at least two mechanisms, a semantic mechanism and a response competition mechanism. PMID- 16035347 TI - A three year follow-up of posterior doxadent restorations. AB - A new direct restorative material, a calcium aluminate cement, was introduced in the year 2000 and sold under the name Doxadent (Doxa AB, Uppsala, Sweden). The aim of this study was to evaluate Doxadent used in class I and II cavities. A total of 63 restorations were evaluated clinically, according to sligthly modified USPHS criteria, at baseline, after 6 mths, 1, 2 and 3 yrs. Four of the patients reported post-operative sensitivity. At 3 yrs, 62 restorations were evaluated. At 6 mths 9.5% non-acceptable Doxadent restorations were observed, 17.5% at 12 mths, 24.2% at 2 yrs and 21% at 3 yrs, which resulted in a cumulative failure frequency of 72.6% at the end of the 3-years for the new restorative material. Main reasons for failure were material or tooth fracture. It can be concluded that Doxadent showed a non-acceptable clinical failure rate as a posterior restorative, especially in class II cavities. PMID- 16035348 TI - Use of abutment-teeth vs. dental implants to support all-ceramic fixed partial dentures: an in-vitro study on fracture strength. AB - The aim of the present in-vitro study was to compare the fracture strength of all ceramic Fixed Partial Dentures supported by tooth-analogues and periodontal membrane with the same supported by dental implants. As ceramics are highly brittle, they cannot withstand deformations of more than 0.1% without fracturing. Hence, when planning an all-ceramic FPD, it is essential to evaluate abutment support, as the fracture strength of all-ceramic constructions depends on the stability of the support to reduce strain in the beam of the prosthesis. The support provided by implants differs, however, from the support provided by natural teeth as the implants are anchored directly in the bone with no intermediate tissue. One question that arises is whether strain and stress in the prosthesis are lower when the prosthesis is loaded on implants compared to natural teeth and hence if all-ceramic FPDs benefit from implant support. Twenty four three-unit all-ceramic FPDs-12 supported by two dental implants and 12 by two tooth-analogues serving as end abutments-were made. All FPDs were subjected to preloading in a preloading procedure and subsequently subjected to load until fracture occurred. Load at fracture were registered and comparisons between the two groups were made. The loads at fracture were statistically significant higher in the group supported by implants compared to the group supported by tooth analogues (p = 0.003). Within the limitations of this in-vitro study, the following conclusions can be drawn: All-ceramic fixed partial dentures can be used in combination with dental implants. The solid support gained from implants might thus be beneficial for the outcome of such treatment due to decreased strain and stress levels in the prosthesis when loaded on implants compared to when loaded on natural teeth. Clinical studies are, however, needed to confirm these findings as there are more factors that influence the final clinical outcome. PMID- 16035349 TI - Silicone sealers, acetic acid vapours and dental erosion: a work-related risk? AB - The aim of the present study was to investigate whether the acetic acid released by some silicone sealers during the curing process poses an increased risk for dental erosion, thus constituting an occupational hazard to exposed individuals. The material comprised 13 individuals (x=30 years, 10 men and 3 women) who had been exposed to an average of 4.2 years' (range 0.6-10 years) of working with silicone. Each had comprehensive medical and dental examinations carried out. A sex- and aged-matched group of 20 healthy, unexposed workers from the same company served as controls for the medical examination, while study models from randomly selected sex- and age-matched individuals were used as controls for assessing the severity of erosion. Using a questionnaire, an assessment of the role of various possible factors related to oral and general health, and to dental erosion in particular, was made for each participant in the exposed group. Clinical examination included recordings of severity of dental erosion, presence of "cuppings", DMFT, salivary secretion rate and buffer capacity, visible plaque index and gingival bleeding index. In addition, bitewing radiographs, study casts and intraoral colour transparencies were obtained for each individual. The severity of dental erosion was significantly higher in those exposed to silicone compared to controls. There was also a significant correlation between the period of exposure to silicone in the workplace and severity of erosion. Medical problems, especially with regard to upper respiratory tract symptoms, were significantly more common among exposed individuals than controls. In conclusion, a relationship between occupational exposure to acetic acid vapours from silicone sealers and development of dental erosion would appear to exist. PMID- 16035350 TI - Dental caries and caries associated factors in Swedish 15-year-olds in relation to immigrant background. AB - The aim of the present study was to evaluate the prevalence of caries and caries associated variables in 15-year-olds in relation to foreign background and to examine differences in the prevalence of caries in immigrant adolescents according to their length of residence in Sweden. All 15-year-old adolescents (n=143) at one public school in the city of Jonkoping, Sweden were asked to participate in the study. The adolescents were divided into two groups according to their background: immigrants and non-immigrants. Data on caries prevalence were extracted from the dental records of the examination made when the participants were 15 years old. The proportions of immigrants and non-immigrants free from carious lesions were equal. Immigrant adolescents, however, had on average more enamel carious lesions. Adolescents born in Sweden of immigrant parents or who had arrived before 1 year of age had a caries prevalence similar to those of non-immigrant adolescents, whereas children who had immigrated to Sweden after 7 years of age had a caries prevalence that was 2-3 times higher. As the caries carious lesions in immigrant adolescents is mainly restricted to the enamel, and possibly reversible, early introduction of preventive programmes seems essential. PMID- 16035351 TI - Dental digital radiography: a survey of quality aspects. AB - The aim was to evaluate the experiences of Swedish general dental practitioners (GDPs) with digital radiography and their opinion on the same, particularly regarding quality issues. A letter was sent to all GDPs in private care in Region Skane, Sweden, asking whether they used digital radiography (n=513). The response rate was 79%. The number of private GDPs who replied that they used digital radiography was 106. The Public Dental Service in Region Skane listed 33 GDPs who worked with digital radiography. Based on these answers, a questionnaire was sent to the GDPs working with digital radiography (n=139). The questionnaire comprised 27 questions about the dentists, the system of intra-oral digital radiography, and the GDPs' experiences of and opinions on issues regarding image quality and quality control. The response rate to the questionnaire was 94%. Almost all, 92%, worked with charge-coupled device (CCD) and complementary metal oxide semiconductor (CMOS) sensors. Most GDPs were satisfied with their digital radiographic system. The majority (65%) experienced problems. Detector failure and trouble with the software were common. The GDPs wrote that they used lower exposure times in digital radiography than traditional film radiography. The estimated reduction in exposure time was said to be between 51% and 75%. Thirty five per cent continued to use film parallel with digital radiography. The answers indicated that less than half of the equipment (40%) underwent quality control. Quality controls, when conducted, were undertaken once or twice a year, mainly by technicians from the companies that had sold the digital equipment. Based on the results of the questionnaire, there seems to be a need to improve the maintenance and the quality of digital radiography. It is also important that the GDPs become more aware of the problems that can occur when a new technique is introduced and that they develop the skills to handle these problems. PMID- 16035352 TI - Automated dynamic liquid-liquid-liquid microextraction followed by high performance liquid chromatography-ultraviolet detection for the determination of phenoxy acid herbicides in environmental waters. AB - Automated dynamic liquid-liquid-liquid microextraction (D-LLLME) controlled by a programmable syringe pump and combined with HPLC-UV was investigated for the extraction and determination of 5 phenoxy acid herbicides in aqueous samples. In the extraction procedure, the acceptor phase was repeatedly withdrawn into and discharged from the hollow fiber by the syringe pump. The repetitive movement of acceptor phase into and out of the hollow fiber channel facilitated the transfer of analytes into donor phase, from the organic phase held in the pore of the fiber. Parameters such as the organic solvent, concentrations of the donor and acceptor phases, plunger movement pattern, speed of agitation and ionic strength of donor phase were evaluated. Good linearity of analytes was achieved in the range of 0.5-500 ng/ml with coefficients of determination, r2 > 0.9994. Good repeatabilities of extraction performance were obtained with relative standard deviations lower than 7.5%. The method provided up-to 490-fold enrichment within 13 min. In addition, the limits of detection (LODs) ranged from 0.1 to 0.4 ng/mL (S/N = 3). D-LLLME was successfully applied for the analysis of phenoxy acid herbicides from real environmental water samples. PMID- 16035353 TI - Microwave assisted extraction followed by gas chromatography with tandem mass spectrometry for the determination of triclosan and two related chlorophenols in sludge and sediments. AB - A procedure for the determination of 2-(2,4-dichlorophenoxy)-5-chlorophenol (Triclosan) and two possible transformation compounds, 2,4-dichlorophenol (2,4 DCP) and 2,4,6-trichlorophenol (2,4,6-TCP), in sludge from sewage treatment plants (STP) and sediments is presented. Extraction was performed using an acetone:methanol (1:1) mixture under the action of a microwave field. The centrifuged supernatant was diluted with a NaOH aqueous solution and twice extracted with n-hexane for removing neutral and basic interferences. The aqueous layer was acidified and processed as a waste water sample. After concentration analytes were silylated and determined by gas chromatography with tandem mass spectrometry (GC-MS/MS). Influence of experimental conditions on the yield of the extraction process and on the efficiency of the further clean-up step was thoroughly evaluated. Performance of MS/MS detection in comparison to single MS is described. Under final working conditions quantification limits between 0.4 and 0.8 ng/g and recoveries from 78% to 106% were obtained. The method was applied to the analysis of several sludge and sediment samples. Only low levels of TCS were detected in some of the sediments; however, all three compounds were found in sludge samples at concentrations ranging from 7 to 316 ng/g, in the case of chlorophenols, and from 420 to 5400 ng/g, for Triclosan. PMID- 16035354 TI - Two-step hollow fiber-based, liquid-phase microextraction combined with high performance liquid chromatography: a new approach to determination of aromatic amines in water. AB - A novel method for the extraction of aromatic amines present in water samples is produced here coupling two-step liquid-phase microextraction with high performance liquid chromatography by using a monolithic column. The hydrophobic porous polypropylene membranes were used as the interface between the donor water sample and the acceptor aqueous solution. In the first step, the analytes were extracted from a sample solution (pH 13) as donor phase into the organic phase, benzyl alcohol-ethyl acetate (80-20%, v/v) immobilized in the pores of a polypropylene dish and further into an acidified acceptor phase (pH 2) inside the polypropylene membrane. This step had about 100% relative recovery with an enrichment factors of over 59.9. For the second step, using a single piece of polypropylene hollow fiber, was shaped with a star liked profile as the acceptor phase. The acceptor solution in the first step was the donor phase for the second step. This solution was adjusted again to pH 13 with NaOH solution. Five microlitres of HCl solution (pH 2) as the acceptor phase was added to the hollow fiber, star liked profile, as the acceptor phase. This acceptor solution, after the second extraction step, was subsequently withdrawn into a micro syringe and directly injected into an HPLC system for analysis. With this two-step microextraction, total enrichment factors of >6000 could be obtained and detection limits of < or = 250.0 ng/l (S/N = 3)could be achieved. The calibration curves for analytes were linear within the range of 20.0 ng/l to 300 microg/l. All expreriments were carried out at room temperature, 22 +/- 0.5 degrees C. PMID- 16035355 TI - Extraction of testosterone and epitestosterone in human urine using aqueous two phase systems of ionic liquid and salt. AB - Based on aqueous two-phase systems (ATPS) consisting of 1-butyl-3 methylimidazolium chloride, a hydrophilic ionic liquid (IL), and K2HPO4, a new and simple extraction technique, coupled with a reversed-phase high performance liquid chromatography (RP-HPLC), was developed for the simultaneous concentration and analysis of testosterone (T) and epitestosterone (ET) in human urine. Under the optimal conditions, the extraction efficiencies for both analytes were 80-90% in a one-step extraction. The method required only 3.0 mL of urine and a single hydrolysis/deproteinization/extraction step followed by direct injection of the IL-rich upper phase into HPLC system for analysis. The method has been satisfactorily applied to the analysis of T and ET in human urine with detection limits of 1 ng/mL and linear ranges of 10-500 ng/mL for both compounds. Compared with conventional liquid-liquid extraction or solid phase extraction, this new method is much "greener" due to no use of volatile organic solvent and low consumption of IL. The proposed extraction technique opens up new possibilities in the separation of other drugs. PMID- 16035356 TI - Combining restricted access material (RAM) and turbulent flow for the rapid on line extraction of the cyclooxygenase-2 inhibitor rofecoxib in plasma samples. AB - Restricted access material (RAM) has been used in the packing of a solid-phase extraction (SPE) column for on-line extractions under turbulent flow conditions. The bio-compatible RAM material works by the principle of size exclusion in addition to conventional reversed-phase chromatography, thereby allowing the extraction and preconcentration of small analyte molecules from biological samples such as plasma. Using small column dimensions (0.76 mm x 50 mm) and a consequently high linear velocity, turbulent flow was achieved during online sample extractions. The improved mass-transfer rate characteristic of turbulent flow allows fast sample cleanup without decreased extraction efficiency. The novel use of the RAM column, connected upstream to a C18 monolithic column, allowed the direct injection, extraction, separation, and MS/MS detection of plasma samples spiked with rofecoxib in a span of 5 min. Calibration curves obtained using this RAM turbulent flow coupled column method showed good linearity (R2 > 0.99) and reproducibility (%RSD < or = 7%). The lower limit of quantitation of rofecoxib in plasma samples was found to be 40 ng/ml. The extraction method showed good recovery of rofecoxib from a plasma matrix with minimal signal loss and robustness after more than 200 plasma injections. PMID- 16035357 TI - Interpretation of the excess adsorption isotherms of organic eluent components on the surface of reversed-phase phenyl modified adsorbents. AB - The adsorption of three organic eluent components (acetonitrile, methanol, and tetrahydrofuran) from water were measured on four phenyl-type bonded phases using the minor disturbance method. The thicknesses of organic layer enriched above the phenyl-type bonded ligands were assessed and interpreted. Acetonitrile and tetrahydrofuran showed multilayer formation while methanol showed monomolecular adsorption. These results were compared to those obtained on alkyl bonded phases. PMID- 16035358 TI - Interactions between sodium dodecyl sulphate and non-ionic cellulose derivatives studied by size exclusion chromatography with online multi-angle light scattering and refractometric detection. AB - The novel approach described allows to characterise the surfactant-polymer interaction under several sodium dodecyl sulphate (SDS) concentrations (0-20 mM) using size exclusion chromatography (SEC) with online multi-angle light scattering (MALS) and refractometric (RI) detection. Three different cellulose derivatives, hydroxypropyl cellulose (HPC), hydroxypropyl methyl cellulose (HPMC) and hydroxyethyl cellulose (HEC), have been studied in solution containing 10 mM NaCl and various concentrations of sodium dodecyl sulphate. It is shown that this approach is well suited for successful application of both Hummel-Dreyer and multi-component light scattering principles and yields reliable molecular masses of both the polymer complex and the polymer itself within the complex, the amount of surfactant bound into the complex as well as appropriate values of the refractive index increment (dn/dc)micro, of both the complex and the polymer in question. The more hydrophobic derivatives HPC and HPMC adsorbed significantly more SDS than HEC. The inter-chain interactions close to critical aggregation concentration (cac) were clearly seen for HPC and HPMC as an almost two-fold average increase in polymer molecular mass contained in the complex. PMID- 16035359 TI - Purification of plasmid DNA vectors by aqueous two-phase extraction and hydrophobic interaction chromatography. AB - The current study explores the possibility of using a polyethyleneglycol(PEG) ammonium sulphate aqueous two-phase system (ATPS) as an early step in a process for the purification of a model 6.1 kbp plasmid DNA (pDNA) vector. Neutralised alkaline lysates were fed directly to ATPS. Conditions were selected to direct pDNA towards the salt-rich bottom phase, so that this stream could be subsequently processed by hydrophobic interaction chromatography (HIC). Screening of the best conditions for ATPS extraction was performed using three PEG molecular weights (300, 400 and 600) and varying the tie-line length, phase volume ratio and lysate load. For a 20% (w/w) lysate load, the best results were obtained with PEG 600 using the shortest tie-line (38.16%, w/w). By further manipulating the system composition along this tie-line in order to obtain a top/bottom phase volume ratio of 9.3 (35%, w/w PEG 600, 6%, w/w NH4)2 SO4), it was possible to recover 100% of pDNA in the bottom phase with a three-fold increase in concentration. Further increase in the lysate load up to 40% (w/w) with this system resulted in a eight-fold increase in pDNA concentration, but with a yield loss of 15%. The ATPS extraction was integrated with HIC and the overall process compared with a previously defined process that uses sequential precipitations with iso-propanol and ammonium sulphate prior to HIC. Although the final yield is lower in the ATPS-based process the purity grade of the final pDNA product is higher. This shows that it is possible to substitute the time consuming two-step precipitation procedure by a simple ATPS extraction. PMID- 16035360 TI - Preparation and characterization of spherical polymer packings from polybutadiene for size-exclusion chromatography. AB - Porous polymer spherical particles for column packings in nonaqueous size exclusion chromatography (SEC) were prepared from 1,2-syndiotactic polybutadiene by suspension and evaporation method. The polymer microbeads obtained were crosslinked by radical reaction between 2-vinyl groups in polybutadiene with ultraviolet radiation, to render them insoluble. These microbeads have wider chromatographic separation width than polystyrene column packings. In addition, the polybutadiene microbeads did not show the excessive retention observed with commercial polystyrene columns for polycyclic aromatic compounds. Therefore, a close correlation between the elution volume and M, for polycyclic aromatic compounds was observed with polybutadiene microbeads columns. PMID- 16035361 TI - High temperature liquid chromatography and liquid chromatography-mass spectroscopy analysis of octylphenol ethoxylates on different stationary phases. AB - Temperature was investigated as active parameter in the liquid chromatography (LC) analysis of octylphenol ethoxylates. Significant differences in selectivity were observed when the oligomers were analyzed by reversed phase LC (RPLC) on silica-, zirconia- and polystyrene/divinylbenzene based stationary phases at low (ambient), medium and elevated temperature with acetonitrile/water as mobile phase. As ascertained by LC-mass spectroscopy (MS), in most cases the elution order of the oligomers was completely reversed comparing ambient and high temperature separations. On a graphitized carbon type column, the selectivity remained unchanged, regardless the analysis temperature. Also in normal phase LC, the elution order remained unaffected by temperature variations both for acetonitrile/water and methanol/water mixtures as mobile phase. Surprisingly, when reversed phase LC on a octadecylsilicagel column at different temperatures was repeated with methanol instead of acetonitrile as mobile phase ingredient, the reversal of elution order did not take place. Results are evaluated in terms of thermodynamic parameters. PMID- 16035363 TI - Adsorption of anions to zirconium(IV) and titanium(IV) chemically immobilized on gel-phase. AB - Adsorption behaviors of 25 anions to zirconium(IV) and titanium(IV) chemically immobilized on a gel-phase were studied by ion chromatography and by batch adsorption experiments. The affinities of Zr(IV) to iminodiacetate as an anchoring group and to anions as a sample are much stronger than those of Ti(IV). On a Zr(IV) column, fourteen anions showed no retention, four anions showed pH dependent retention in a low pH region, and seven anions were irreversibly adsorbed at pH <7. In the last group, fluoride was adsorbed both by the ligand exchange mechanism and the addition mechanism, while phosphate, arsenate and selenite were only by the ligand exchange mechanism. The structures of the adsorbed species are discussed. PMID- 16035362 TI - Application of preparative high-speed counter-current chromatography for isolation and separation of schizandrin and gomisin A from Schisandra chinensis. AB - Following an initial cleaning-up step on the D101 macroporous resin, a preparative high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (1:0.9:0.9:1, v/v) was used to isolate and separate schizandrin and gomisin A from Schisandra chinensis. A total of 107 mg schizandrin and 36 mg gomisin A with purities of 99.5% and 99.1% were obtained from 400 mg crude extract in one-step elution and less than 3 h, and the structure identification was performed by UV, IR, MS, 1H NMR and 13C NMR. PMID- 16035364 TI - Epoxidized soy bean oil migrating from the gaskets of lids into food packed in glass jars. Analysis by on-line liquid chromatography-gas chromatography. AB - The migration of epoxidized soy bean oil (ESBO) from the gasket in the lids of glass jars into foods, particularly those rich in edible oil, often far exceeds the legal limit (60 mg/kg). ESBO was determined through a methyl ester isomer of diepoxy linoleic acid. Transesterification occurred directly in the homogenized food. From the extracted methyl esters, the diepoxy components were isolated by normal-phase LC and transferred on-line to gas chromatography with flame ionization detection using the on-column interface in the concurrent solvent evaporation mode. The method involves verification elements to ensure the reliability of the results for every sample analyzed. The detection limit is 2-5 mg/kg, depending on the food. Uncertainty of the procedure is below 10%. PMID- 16035365 TI - Effect of ionic additives on the elution of sodium aryl sulfonates in supercritical fluid chromatography. AB - Addition of a small amount of polar solvent (i.e., modifier) to CO2 in packed column supercritical fluid chromatography (SFC) has shown major improvements in both polar analyte solubility and interaction of the polar analyte with the stationary phase. Recently, the addition of an ionic component (i.e., additive) to the primary modifier by one of us has been shown to extend even further the application of SFC to polar analytes. In this work, the effect of various ionic additives on the elution of ionic compounds, such as sodium 4-dodecylbenzene sulfonate and sodium 4-octylbenene sulfonate, has been studied. The additives were lithium acetate, ammonium acetate, tetramethylammonium acetate, tetrabutylammonium acetate, and ammonium chloride dissolved in methanol. Three stationary phases with different degrees of deactivation were considered: conventional cyanopropyl, deltabond cyanopropyl, and bare silica. The effect of additive concentration and additive functionality on analyte retention was investigated. Sodium 4-dodecylbenzene sulfonate was successfully eluted using all the additives with good peak shape under isocratic/isobaric/isothermal conditions. Different additives, however, yielded different retention times and in some cases different peak shapes. PMID- 16035366 TI - Separation of enantiomers in capillary electrophoresis with contactless conductivity detection. AB - Contactless conductivity detection is successfully demonstrated for the enantiomeric separation of basic drugs and amino acids in capillary electrophoresis (CE). Derivatization of the compounds or the addition of a visualization agent as for indirect optical detection schemes were not needed. Non-charged chiral selectors were employed, hydroxypropylated cyclodextrin (CD) for the more lipophilic basic drugs and 18-crown-6-tetracarboxylic acid (18C6H4) for the amino acids. Acidic buffer solutions based on lactic or citric acid were used. The detection limits were determined as 0.3 microM for pseudoephedrine as an example of a basic drug and were in the range from 2.5 to 20 microM for the amino acids. PMID- 16035367 TI - Kinetic study of cytochrome P450 3A4 activity on warfarin by capillary electrophoresis with fluorescence detection. AB - The use of capillary electrophoresis (CE) for the determination of cytochrome P450 3A4 (CYP3A4) activity with R-warfarin as a substrate was investigated. CYP3A4 activity was determined by the quantitation of the product, 10 hydroxywarfarin, based on separation by CE. The separation conditions were as follows: capillary, 80.5 cm (75 microm i.d., 60 cm effective length); 50 mM sodium phosphate buffer (pH 6.5); 23 kV (90 microA) applied voltage; fluorescence detection, excitation wavelength, 310 nm, emission wavelength, 418 nm; capillary temperature, 37 degrees C. With the developed CYP3A4 activity assay and the Lineweaver-Burk equation, the Michaelis-Menten parameters Km and Vmax for formation of 10-hydroxywarfarin from R-warfarin in the presence of CYP3A4 were calculated to be 166 +/- 12 microM and 713 +/- 14 pmol/min/nmol (or 91.4 pmol/min/mg) CYP3A4, respectively. PMID- 16035368 TI - [Recommendations for the prevention of cardiovascular diseases in clinical practice]. AB - These recommendations are largely based on the Executive Summary of the "European Guidelines on Cardiovascular Disease Prevention in Clinical Practice" proposed by the "Third Joint Task Force of European and other Societies on Cardiovascular Disease Prevention in Clinical Practice". The model used to assess the overall risk was adapted for Belgium. Otherwise, very few things were changed from the Executive Summary of the European Guidelines. PMID- 16035369 TI - [Bioequivalence in vitro evaluation of some antibacterial generic dosage forms]. AB - In this work, bioequivalence between generic and corresponding original brand name dosage forms of some antibacterial drugs, frequently prescribed in developing countries, have been examined using in vitro dissolution testing. For this purpose, tablet or hard capsule formulations of five active substances (amoxycillin, ampicillin, co-trimoxazole (sulphamethoxazole/trimethoprim), metronidazole and penicillin V) have been retained. For each active substance, batch samples of three generic and one test formulations have been submitted to the pharmaceutical quality control and dissolution testing. Results obtained have shown that all samples examined met the specifications of quality edited by the pharmacopeias. On the other hand, interchangeability between generic and corresponding test formulations should be possible since their dissolution profiles are superposables enough. PMID- 16035370 TI - [Pharmaceutical forms during the Enlightenment]. PMID- 16035371 TI - [Pharmaceutical containers and their technological feature at the end of the middle ages]. PMID- 16035372 TI - Ratio of lathosterol to campesterol in serum predicts the cholesterol-lowering effect of sitostanol-supplemented margarine. AB - OBJECTIVE: Plant sterol/stanol margarines are recommended as a lipid-lowering dietary supplement in the treatment of hypercholesterolemia. Parameters predicting the individual cholesterol-lowering effect have not been elucidated so far. Therefore, we investigated the responsiveness to sitostanol-supplemented margarine in a specially selected population. METHODS AND RESULTS: From a total number of 137 male subjects with hypercholesterolemia, eight subjects with the lowest and eight subjects with the highest ratios of lathosterol to campesterol in serum were included in the study. They received 1 g sitostanol-supplemented margarine b.i.d. for four weeks. Serum lipoproteins, the cholesterol precursor lathosterol, the plant sterols campesterol and sitosterol were measured. Subjects with a low ratio of lathosterol to campesterol had a significant decrease of serum total cholesterol (-14.2%; p < 0.01) and LDL cholesterol (-13.8%; p < 0.01; responder). In subjects with a high ratio there was no significant change in total cholesterol and LDL cholesterol (2.2 and 4.3%; non-responder). CONCLUSION: The ratio of serum lathosterol to campesterol predicts the reduction of total cholesterol and LDL cholesterol during administration of sitostanol-supplemented margarine in patients with mild hypercholesterolemia. PMID- 16035373 TI - Factors influencing neonatal therapeutic effect of anti-MRSA drugs. AB - OBJECTIVE: Factors influencing the neonatal therapeutic effect of anti-MRSA (methicillin-resistant Staphylococcus aureus) drugs are investigated. METHODS: This study took place over a two-year period from April 1998 to March 2000. We calculated the non-adjusted odds ratio for each influential factor to determine the therapeutic effect of anti-MRSA drugs. RESULTS: Significant factors for therapeutic effect were found to be platelet count, urea nitrogen, creatinine, and CRP, each measured before starting administration of anti-MRSA drugs; whether blood drug concentration was measured; and whether pneumonia or septicemia was present. There was a tendency where a better therapeutic effect was gained when the total protein and albumin values were high. We applied multivariate logistic regression analysis to these factors, and found the following independent significant factors: CRP (odds ratio (OR) = 1.582), albumin (OR = 3.079), Cre (OR -0.213), whether blood drug concentration was measured (OR = 3.767), and presence of pneumonia or septicemia (OR = 0.216). This result suggests that consideration should be given to these five important factors when treating MRSA patients. PMID- 16035374 TI - Effect of a dietary supplement containing probiotic bacteria plus vitamins and minerals on common cold infections and cellular immune parameters. AB - OBJECTIVE: A randomized, double-blind, placebo-controlled intervention study was carried out in order to investigate whether consumption of a dietary supplement containing probiotic bacteria plus vitamins and minerals over a period of at least three months in winter/spring affects the duration, frequency, and severity of symptoms of naturally acquired common cold infections as well as cellular immune parameters. METHODS: 477 healthy men and women (aged 36 +/- 13, mean +/- SD) who had not been vaccinated against influenza were randomly assigned to a group who received daily the probiotic multivitamin and mineral supplement (verum) or a placebo, for three (n = 239) or for 5.5 months (n = 238). Cellular immune response was determined in 60 participants per study group by flow cytometry before and after 14 days of supplementation. RESULTS: The incidence of respiratory tract infections regarded as being virally induced was 13.6% lower in the verum group compared to the placebo group (p = 0.07). During respiratory tract infection episodes, the subjects recorded common cold and influenza-like symptoms daily. All symptoms were reduced in the verum group. We found a relative reduction of 19% in the total symptom score (p = 0.12), 25% in influenza symptoms (p = 0.09), and 54% in the number of days with fever (p = 0.03). The duration of these infections was not affected. Leukocytes, lymphocytes, in particular T lymphocytes including CD4+ and CD8+ cells, as well as monocytes were significantly higher increased in the verum group, during the first 14 days of supplementation compared to placebo. CONCLUSION: These data indicate that the intake of a dietary supplement containing probiotic bacteria plus vitamins and minerals during a period of at least three months in winter/spring may reduce the incidence and the severity of symptoms in common cold infections in otherwise healthy adults. This may be due to stimulated cellular immunity. PMID- 16035375 TI - Rate-limiting biotransformation of triamterene is mediated by CYP1A2. AB - OBJECTIVE: Triamterene (TA), a potassium-sparing diuretic, is extensively metabolized by hydroxylation in 4'-position and subsequent conjugation by cytosolic sulfotransferases. To identify the cytochrome P450 enzyme(s) catalyzing hydroxylation of triamterene (the rate-limiting step in the formation of the sulfate ester (STA)), in vitro incubation studies were performed with human liver microsomes. METHODS: Initial rates of TA hydroxylation (0 - 300 microM) were determined during a ten-minute-incubation period with liver microsomes of two donors. The role of individual CYP enzymes was determined by pre-incubation with selective inhibitors/alternative substrates. Vice versa, the effect of TA (0 - 500 microM) on 3-demethylation of caffeine (0 - 1,000 microM) was assessed. Metabolite concentrations were estimated by reversed-phase HPLC methods. RESULTS: TA Km values without inhibitors were 60 and 142 microM, Vmax was 177 and 220 pmol/min/mg protein, respectively. Mean inhibitor induced changes of 4'-hydroxy TA formation were as follows: Furafylline 25 microM (CYP1A2), complete inhibition (-100%); omeprazole 250 microM (CYP1A2 inhibitor/CYP2C 19 substrate), -30%; coumarin 25 microM (CYP2A6), -11%; quinidine 25 microM (CYP2D6), -9%; ketoconazole 25 microM (CYP3A), -18%; and erythromycin 250 microM (CYP3A), -8%. In the reverse inhibition studies, TA competitively inhibited caffeine 3 demethylation with Ki values of 65 and 111 microM, respectively. CONCLUSION: 4' hydroxylation of TA in humans appears to be mediated exclusively by CYP1A2. Inhibition or induction of CYP1A2 will change the time course of both TA and its active phase-II metabolite. The net pharmacodynamic effect of such changes is difficult to predict and needs to be evaluated in clinical studies. PMID- 16035376 TI - Comparison of two low-molecular weight heparins (LMWHs), tinzaparin and bemiparin, during hemodialysis. AB - The anticoagulant effects of two LMWHs, tinzaparin sodium and bemiparin sodium, were compared during two study dialysis (SD) sessions (SD1 with tinzaparin and SD2 with bemiparin) in ten chronic hemodialysis patients. Prior to SD1, patients had received 3,500 IU of tinzaparin in each dialysis session for two weeks. After SD1, they were switched to 3,500 IU of bemiparin for two weeks and the second study dialysis session (SD2) was carried out. Patients used the same dialyzers during the whole study period and no changes were made in any of the other hemodialysis parameters. Blood samples for the measurement of PT, aPTT and anti Xa activity were taken before the study dialysis sessions (sample 0), and again at two and four hours after the initiation of dialysis (samples 2-h and 4-h, respectively). PT values showed no change between the SD1 and SD2 sessions, but aPTT values were significantly higher in the 2-h and 4-h samples of the SD1 session compared to corresponding values in SD2 samples (p < 0.005). Anti-Xa activity was higher in the 2-h and 4-h samples of the SD2 session compared to the SD 1 session (p = 0.005 and p = 0.012, respectively). At four hours, only patients receiving bemiparin had a mean anti-Xa activity higher than 0.40 IU/mg (mean 0.51 +/- 0.76). However, fibrin/clot formation in the extracorporeal circuit during each study period as assessed by visual inspection, did not differ significantly between sessions. It is concluded that the anticoagulant profile of bemiparin sodium during hemodialysis treatment is superior to that of tinzaparin. PMID- 16035377 TI - Prescribing practices in German and Swiss psychiatric university and in non university hospitals: national differences. AB - OBJECTIVE: There are great variations between hospitals in the way drugs are prescribed, and these variations may be due to multiple factors such as local prescribing traditions, pharmacoeconomic considerations, drug availability, regional differences of population, disease prevalence etc. Available studies on prescribing habits, apart from studies performed in a unique center, have until now been mainly restricted to single countries or regions and the comparisons across countries or regions have often been limited by the use of diverse methodologies and definitions. The aim of the present study was to compare drug prescriptions between German and Swiss psychiatric services with regard to their preference of newer psychotropics. MATERIAL AND METHODS: Five psychiatric hospitals, associated to the AMSP project, were chosen to represent Swiss and German clinics, university and non-university settings. Data were available from one index day on 572 patients and 1,745 prescriptions. The comparisons were adjusted for age and gender. RESULTS: There was a significant difference (p < 0.001) with regard to the prescription of newer antidepressants (NAD), Swiss clinicians giving proportionally more (65.2%) than the German psychiatrists (48.3%). No significant difference was, on the other hand, found as to the proportion of atypical antipsychotics, the lack of difference being due to the higher proportion of clozapine among the atypical antipsychotics in Germany. CONCLUSION: There seems, therefore, to be a higher propensity for Swiss hospital psychiatrists to prescribe newer antidepressants. This seems to be due to national or regional prescribing traditions. Further studies are needed to investigate the economical influences on antidepressant prescribing in Swiss and German clinics. PMID- 16035378 TI - Comparative bioavailability of 875 mg amoxicillin tablets in healthy human volunteers. AB - OBJECTIVE: To compare the bioavailability of amoxicillin 875 mg tablets (EMS Sigma Pharma used as test formulation) and Amoxil BD 875 mg tablets (GlaxoSmithKline used as reference formulation) in 26 healthy volunteers. MATERIAL AND METHODS: 26 healthy volunteers (13 males and 13 females) received each formulation in an open, 2 x 2 crossover, randomized study with seven days of washout period between doses. Plasma samples were obtained over a 12-hour interval after administration. Plasmatic amoxicillin concentrations were obtained by combined reversed-phase liquid chromatography and mass spectrometry with positive ion electrospray ionization using the select ion monitoring method. AUC was calculated by the trapezoidal rule extrapolation method. Cmax and tmax were compiled from the plasmatic concentration-time data. Analysis of variance was carried out using logarithmically transformed AUC0-inf, AUC0-12 h, Cmax and untransformed tmax. RESULTS: The mean values (+/- SD) for AUC0-12 h (microg x h x ml(-1)), AUC0-inf (microg x h x ml(-1)), Cmax (microg x ml(-1)), t1/2 (h) and tmax (h), were, respectively: 55.42 (+/- 16.85), 55.42 (+/- 16.85), 18.59 (+/- 6.3), 1.49 (+/- 1.57) and 2.04 (+/- 0.75) concerning the test formulation, and 51.11 (+/- 18.9), 51.29 (+/- 19.12), 17.83 (+/- 5.86), 1.52 (+/- 1.31) and 2.02 (+/- 0.87) concerning the reference formulation. Confidence intervals (90%) of amoxicillin means of AUC0-12 h and Cmax ratios (test/reference) were: 0.961-1.149 and 0.914-1.142, respectively, agreeing with the bioequivalence criteria established by the Brazilian National Health Surveillance Agency. CONCLUSION: Both formulations were bioequivalent based on both the rate and extent of absorption. PMID- 16035379 TI - Breast-conserving therapy in breast cancer patients--a 12-year experience. AB - INTRODUCTION: Twenty years ago prospective randomised controlled trials were initiated to compare conservative breast surgery plus radiation with radical mastectomy in the treatment of early-stage breast cancer. The results have shown no survival advantage for mastectomy over breast-conserving therapy (BCT). However, local recurrence of cancer after BCT has been reported to be as high as 14%, necessitating salvage mastectomy. METHODS: This retrospective study was performed on 165 breast cancer patients undergoing BCT in the 12 years up to August 2002. Resection and intraoperative cytological assessment were used to achieve clear excision margins. Adjuvant therapy (hormones, chemotherapy) was undertaken, and the incidence and times of local recurrence and distant metastases were recorded. RESULTS: Ninety-four per cent of patients had clear margins at the initial operation. This was achieved irrespective of ductal carcinoma in situ alone or surrounding the cancer in 62% of cases. At a median follow-up of 65 months one patient developed local recurrence (LR) in the breast synchronously with distant relapse. Two patients had non-nodal axillary recurrences but no patient suffered LR in isolation in the treated breast. CONCLUSION: BCT is a safe alternative to mastectomy provided that the tumour is completely excised. The segment containing the cancer should be resected from the nipple to the periphery of the breast. Intraoperative cytological assessment helps to ensure clear margins. Reexcision is recommended for patients with close/involved margins. PMID- 16035380 TI - Spectrum of causes of intestinal obstruction in adult Nigerian patients. AB - OBJECTIVE: To present changes in the cause of intestinal obstruction in an African setting. DESIGN: Consecutive cases of acute intestinal obstruction from 1985 to 1994. SETTING: Obafemi Awolowo University Teaching Hospitals Complex, Ile Ife, Nigeria. SUBJECTS: Adult patients with clinical and radiological evidence of intestinal obstruction. RESULTS: There were 99 patients (60 males) aged 15 - 101 years (mean age 45 years). The majority of patients were young and middle-aged adults. Main causes of obstruction included adhesion (N = 44), volvulus (N =15), external hernias (N = 11), colorectal carcinoma (N = 10) and intussusception (N = 8). Approximately two-thirds of patients (28/44) with adhesion had had previous abdominal operations. The overall mortality was 14%, mainly owing to strangulation obstruction and colonic malignancy. CONCLUSIONS: The increasing role of adhesions as a cause of acute intestinal obstruction demands greater need for routine preventive measures against adhesion formation. PMID- 16035381 TI - Primary hydatid cysts of the pancreas. AB - Pancreatic involvement by hydatid disease is uncommon. Establishing a precise diagnosis may be difficult because the presenting symptoms and findings of investigations may be similar to other more commonly encountered cystic lesions of the pancreas. We report 4 patients with primary hydatid cysts in the head of the pancreas. The records of all patients treated for hydatid disease from 1980 to 2000 were reviewed. During the study period a total of 280 patients were treated, 4 of whom had hydatid disease involving only the pancreas. The 4 patients (3 women, 1 man) ranged in age from 17 to 60 years. Three patients presented with jaundice, abdominal pain and weight loss, 2 with hepatomegaly and 1 with an epigastric mass. All 4 lesions involved the head of the pancreas and ranged in size from 3 to 10 cm in diameter. In 2 patients the investigations incorrectly suggested a cystic tumour and both underwent pancreaticoduodenectomy. In 2 patients the correct diagnosis allowed local excision to be performed. Hydatid cyst is a rare cause of a cystic mass in the head of the pancreas, but should be included in the differential diagnosis of cystic lesions of the pancreas, especially in endemic areas. PMID- 16035382 TI - Traumatic abdominal wall hernia--four cases and a review of the literature. AB - OBJECTIVE: To review blunt traumatic abdominal wall hernias (TAWHs) in our institution. METHOD: Retrospective review of blunt abdominal trauma cases over a 6-month period. RESULTS: Four patients with TAWH were identified. The mean age was 36 years. Three had been involved in vehicular collisions, and 1 had been assaulted with a large stone. All were diagnosed on presentation, 3 by computed tomography scan and 1 clinically. Two were repaired as emergencies, and 1 was repaired after 4 months. The 4th patient refused surgery. CONCLUSION: This uncommon injury requires a high index of suspicion and a low threshold for intervention. CT scan offers the best imaging potential. PMID- 16035383 TI - Strategic planning and the changing nature of effective surgical leadership. PMID- 16035384 TI - Blunt abdominal trauma in Zaria, Nigeria. PMID- 16035385 TI - [Genetic counselling]. AB - 'Genetic counselling is the process by which patients or relatives at risk of a disorder that may be hereditary are advised of the consequences of the disorder, the probability of developing and transmitting it and of the ways in which this may be prevented or ameliorated.' The genetic counsellor will discuss the genetic basis and medical facts and also possible personal, familial, social and insurance implications of the respective disorder with the consultand. Further diagnostic investigations, such as genetic tests, may be helpful or necessary for risk predictions or may be required to reach a firm diagnosis. In asymptomatic individuals at risk for a late-onset genetic disorder genetic counselling is obligatory for predictive genetic testing. Genetic counselling also should be offered to symptomatic patients before a diagnostic test is performed. Thereby the patients might become aware in time about the personal, familial and social consequences of the test result that may exceed that of their actual illness. Due to medical confidentiality the geneticist is not allowed to contact other family members and to inform them about a familial disorder and warn them about their own genetic risk. This is the concern of the consultand. Various aspects make up genetic counselling as a specific process which concerns genetic diagnosis, risk estimation and accurate information on medical and genetic facts, but also has a supportive role ensuring that consultands may benefit from given advice and possible preventive measures. PMID- 16035386 TI - [Genetic causes of mental retardation]. AB - Mental retardation (MR) is defined as congenital or early onset lifelong impairment of cognitive and adaptive functioning (IQ < 70). It effects approximately 3% of the Western population. The causes are heterogenous. Numerical or structural chromosome abnormalities are responsible for 10-20% of the mild cases (MMR) and 40% of the severe cases (SMR). Among them Down syndrome represents the most frequent chromosome aberration and the most frequent defined MR syndrome. Gonosomal aberrations do not coincide with MR, as long as only one gonosome is lost or gained. Nearly all unbalanced structural autosomal aberrations cause SMR. Recent studies suggested that sub-microscopic chromosomal microdeletions or subtelomeric rearrangements account for approximately 10% of the undiagnosed cases. They represent a group of newly defined disorders. Single gene mutations are responsible for > 1200 known syndromal conditions with MR. But only few causative genes have been identified as yet. However, an increasing number of genes causing X-linked mental retardation (XLMR) have been localized and cloned, namely 38 genes of the 136 known syndromic conditions and 19 for the non-syndromic conditions. XLMR explains the 20 % excess of males over females. Despite the increasing knowledge about the causes of MR, about half of the cases remain undiagnosed. Guidelines for the diagnostic procedure in children with MR have been proposed. PMID- 16035387 TI - [Cystic fibrosis--the most frequent life-shortening autosomal recessive disease]. AB - Cystic Fibrosis (CF) is one of the most frequent genetic diseases in the white populations of Europe and North America. Its clinical manifestations are highly variable, ranging from a characteristic life-shortening pathology of the lungs and the pancreas in classical CF to symptoms mainly restricted to male sterility in patients with congenital bilateral aplasia of the vasa deferentia (CBAVD). The genetic basis of CF is mutations in both copies of the CFTR gene, which codes for an ion channel. Even though one single mutation, deltaF508, is responsible for two-thirds of all mutated CFTR alleles, a total of over 1000 CFTR mutations have been described, whose relative frequencies vary between different ethnic groups, and whose biochemical consequences are correlated to the clinical manifestations of the disease. Since 4% of white Europeans and North Americans are healthy heterozygous carriers of CF, the molecular diagnosis of this disease, offered in the context of genetic counselling by a specialist in human genetics, is of great importance. PMID- 16035388 TI - [Neurofibromatosis: the most frequent hereditary tumor predisposition syndrome]. AB - With an incidence of one in 3000 to 4000 individuals, neurofibromatosis type 1 (NF1) is one of the most common autosomal dominant genetic diseases and very likely the most frequent disorder with increased cancer risk. Approximately fifty percent of all patients are familial cases and the remaining half consists of sporadic cases with no affected parent. The hallmark clinical features present in over 90% of all patients are cafe-au-lait spots and neurofibromas. However, the disorder should not be underestimated as a "mere cosmetic problem", since NF1 patients are at increased risk to also develop malignant tumours, such as malignant peripheral nerve sheath tumours (MPNST), juvenile myelomonocytic leukaemia (JMML), optic glioma and pheochomocytoma. Renovascular disease represents an additional risk factor for NF1 patients. The NF1 gene is a classic example for a tumour suppressor gene. It functions as a negative regulator of the protooncogene Ras. This function explains well its involvement in tumour formation. During the last 15 years, since the cloning of the gene, enormous progress has been made towards a better understanding of the natural history of the disorder. However, it cannot be said if and when a cure of the disorder will be possible. Great advantages have been achieved in the monitoring and management of several NF1 complications, for instance in the treatment of tibia pseudarthrosis and optic gliomas. Owing to the technical improvements of the approaches applied to identify NF1-mutations molecular-genetic testing with high mutation detection rates may help nowadays in patients in which the clinical diagnosis may not readily be established, such as in young children or atypical cases. A greater awareness of the complications and the different expression forms of NF1 and NF2 on the part of all types of physicians will further help to offer all patients adequate and timely counselling and treatment. The establishment of multi-disciplinary counselling and treatment centres for neurofibromatosis could be an important step towards a better management of NF1 and NF2 patients. PMID- 16035389 TI - [Tumor cytogenetics]. AB - Cytogenetic analysis by classical chromosome banding, in combination with different fluorescence in situ hybridization (FISH) based methods, including comparative genomic hybridization and multicolour FISH, provides important information concerning the diagnosis, staging and prognosis of, as well as the planning of therapeutic intervention against, leukemias, malignant lymphomas and solid tumors, and may be helpful in monitoring the course of the disease. Cytogenetic methods identify primary chromosome anomalies that are causally involved in the emergence of the disease and its histopathological subtype, as well as secondary anomalies which have an effect upon the course of the disease and the responsiveness to therapy. Tumor cytogenetics is an independent discipline, and by no means only a diagnostic service for, or subdiscipline of, haematological, oncological and pathological fields. It is important, however, to point out that tumor cytogenetics is a cooperation-orientated and interdisciplinary science that deals with its own original questions in collaboration with clinicians, pathologists and molecular geneticists. PMID- 16035390 TI - The Golden Agers and Tick-borne encephalitis. Conference report and position paper of the International Scientific Working Group on Tick-borne encephalitis. AB - The 7th meeting of the ISW TBE had the main topic "Tick-borne encephalitis in the Golden Agers". Data from 14 European countries were presented about incidence and clinical course of Tick borne encephalitis (TBE) in general and especially in the population over 50 years of age. With age immunity is impaired quantitatively and qualitatively, the reactions to vaccinations are generally slower, antibody titres reach lower values and decrease earlier. The incidence of the disease is increasing with age, also the clinical course is more severe, they suffer significantly more sequelae, need a longer rehabilitation and have a higher case fatality. Vaccination as the only efficient protection is needed in endemic areas, considering that mobility has increased very much. For the age group over 50 years regular booster vaccinations according to the recommended vaccination intervals or even shorter intervals are most important. PMID- 16035391 TI - Searching for genes in diabetes and the metabolic syndrome. AB - Evidence for a genetic basis for type 2 diabetes and the metabolic syndrome has been derived from studies of families, twins and populations with genetic admixture. Identification of genes associated with disease pathogenesis is now underway using techniques such as genome scanning by positional cloning and the candidate gene approach. Genome scanning in several different ethnic groups has identified chromosome regions harbouring type 2 diabetes susceptibility genes such as the novel gene, calpain 10 (CAPN10). The hepatic nuclear factor 4alpha (HNF4alpha) gene partly explains the linkage peak on chromosome 20, while the upstream transcription factor (USF1) is associated with familial combined hyperlipidaemia (FCHL) and maps close to the type 2 diabetes associated 1q peak. Peroxisome proliferator-activated receptor gamma (PPARgamma) was identified as a candidate gene based on its biology. A Pro12Ala variant of this gene has been associated with an increased risk of type 2 diabetes. Many genes accounting for monogenic forms of diabetes have been identified--such as maturity onset diabetes of the young (MODY); glucokinase (GCK) and HNF1alpha mutations being the most common causes of MODY. GCK variants result in 'mild' diabetes or impaired glucose tolerance (IGT) and relatively few cardiovascular complications, while HNF1alpha associated MODY is more typical of type 2 diabetes, frequently being treated with sulphonylureas or insulin and resulting in microvascular complications. Testing for single gene disorders associated with type 2 diabetes and obesity may determine cause, prognosis and appropriate treatment; however, for the more common polygenic diseases this is not the case. In type 2 diabetes, molecular genetics has the potential to enhance understanding of disease pathogenesis, and help formulate preventative and treatment strategies. PMID- 16035392 TI - Dysfunctional fat cells, lipotoxicity and type 2 diabetes. AB - Type 2 diabetes is characterized by insulin resistance and impaired insulin secretion. Considerable evidence implicates altered fat topography and defects in adipocyte metabolism in the pathogenesis of type 2 diabetes. In individuals who develop type 2 diabetes, fat cells tend to be enlarged. Enlarged fat cells are resistant to the antilipolytic effects of insulin, leading to day-long elevated plasma free fatty acid (FFA) levels. Chronically increased plasma FFA stimulates gluconeogenesis, induces hepatic and muscle insulin resistance, and impairs insulin secretion in genetically predisposed individuals. These FFA-induced disturbances are referred to as lipotoxicity. Enlarged fat cells also have diminished capacity to store fat. When adipocyte storage capacity is exceeded, lipid 'overflows' into muscle and liver, and possibly the beta-cells of the pancreas, exacerbating insulin resistance and further impairing insulin secretion. In addition, dysfunctional fat cells produce excessive amounts of insulin resistance-inducing, inflammatory and atherosclerosis-provoking cytokines, and fail to secrete normal amounts of insulin-sensitizing cytokines. As more evidence emerges, there is a stronger case for targeting adipose tissue in the treatment of type 2 diabetes. Peroxisome-proliferator activated receptor gamma (PPARgamma) agonists, for example the thiazolidinediones, redistribute fat within the body (decrease visceral and hepatic fat; increase subcutaneous fat) and have been shown to enhance adipocyte insulin sensitivity, inhibit lipolysis, reduce plasma FFA and favourably influence the production of adipocytokines. This article examines in detail the role of adipose tissue in the pathogenesis of type 2 diabetes and highlights the potential of PPAR agonists to improve the management of patients with the condition. PMID- 16035393 TI - Inflammation, dyslipidaemia, diabetes and PPars: pharmacological interest of dual PPARalpha and PPARgamma agonists. AB - Cardiovascular disease (CVD) remains the leading cause of mortality in developed countries. Several risk factors are associated with CVD, including type 2 diabetes, obesity, insulin resistance, dyslipidaemia and hypertension. Different pharmacological therapies have been developed to control these risk factors. Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors, which belong to the nuclear receptor superfamily that controls lipid and glucose metabolism as well as inflammatory risk factors for CVD. PPARalpha agonists, such as the fibrates, correct dyslipidaemia, thus decreasing CVD risk. PPARgamma agonists, such as the glitazones, increase insulin sensitivity and decrease plasma glucose levels in patients with diabetes. Moreover, both PPARalpha and PPARgamma agonists exert anti-inflammatory activities in liver, adipose and vascular tissues. In this review, we focus on the mode of action of PPARalpha and PPARalpha agonists, illustrating the potential of the newly developed dual PPAR agonists for the treatment of global risk in patients with the metabolic syndrome or type 2 diabetes. PMID- 16035394 TI - A review of the efficacy of rosuvastatin in patients with type 2 diabetes. AB - It has been estimated that 92% of individuals with type 2 diabetes, without cardiovascular disease (CVD), have a dyslipidaemic profile. Several guidelines on cardiovascular risk now recommend that patients with diabetes should be considered at high risk of CVD and should thus receive lipid-lowering therapy to reduce low-density lipoprotein cholesterol (LDL-C) to below 2.5 mmol/L. Since their introduction in 1987, statins have revolutionized the management of CVD. The most recent statin to be introduced, rosuvastatin, has been shown to be the most effective at lowering LDL-C, as well as consistently raising HDL-C across the 10-40 mg dose range. This has been confirmed by many studies, including the Measuring Effective Reductions in Cholesterol Using Rosuvastatin Therapy (MERCURY I) study in which rosuvastatin 10 mg was shown to be more effective than commonly used doses of other statins, both for LDL-C reduction and achieving treatment target goals. The effectiveness of rosuvastatin has also been studied in type 2 diabetes patients in three studies: the URANUS (Use of Rosuvastatin vs. Atorvastatin iN type 2 diabetes mellitUS), ANDROMEDA (A raNdomized, Double-blind study to compare Rosuvastatin [10 & 20 mg] and atOrvastatin [10 & 20 Mg] in patiEnts with type II DiAbetes) and CORALL (COmpare Rosuvastatin [10-40 mg] with Atorvastatin [20-80 mg] on apo B/apo A-1 ratio in patients with type 2 diabetes meLLitus and dyslipidaemia) studies. URANUS and ANDROMEDA showed rosuvastatin to be more effective than atorvastatin at reducing LDL-C and achieving treatment target goals. CORALL demonstrated rosuvastatin 10, 20 and 40 mg to be more effective at lowering LDL-C than 20, 40 and 80 mg of atorvastatin, respectively. Ongoing studies will evaluate whether these properties of rosuvastatin translate into beneficial effects on atherosclerosis and significant reductions in cardiovascular events. PMID- 16035395 TI - Statins: potent vascular anti-inflammatory agents. AB - Atherosclerosis is a multifactorial condition that can result in cardiovascular disease. Statin therapy is thought to mediate cardioprotective effects that influence endothelial function, inflammatory responses, plaque stability and thrombus formation, processes involved in atherosclerosis. Although reduction in low-density lipoprotein cholesterol (LDL-C) potentially plays a role in all of these effects, several lines of evidence also implicate nonlipidmediated 'pleiotropic' effects. For example, statin therapy confers a lower risk for coronary heart disease than placebo in patients with comparable serum cholesterol levels, and confers a greater magnitude of clinical benefit than expected based on LDL-C levels alone. Moreover, while nonstatin lipid-lowering therapy does not necessarily reduce stroke risk, statins have shown a significant reduction in stroke. Statins exert their pleiotropic effects, in part, by improving endothelial function via up-regulation of endothelial nitric oxide synthase enzyme activity. Markers of inflammation such as high sensitivity C-reactive protein have been also shown to add further prognostic information about patients at risk of cardiovascular disease who may benefit from statin therapy. Further studies are still needed to determine whether statins have direct effects on inflammatory pathways. PMID- 16035396 TI - Cardioprotective and other emerging effects of statins. AB - The lipid-lowering properties of statins are accompanied by a number of other cardioprotective effects. These 'pleiotropic' actions affect almost the entire process of atherogenesis, from initial endothelial injury to the moment of plaque rupture and thrombosis. This paper discusses the nonlipid-lowering effects of statins that affect the initiation, progression, regression and repair of atherosclerosis lesions, as well as factors that affect plaque instability. The emergent anti-inflammatory, antioxidant and immunomodulatory properties of statins are extensive and diverse. Many of these properties, which are independent of cholesterol synthesis inhibition, help to reduce the ischaemic burden on the cardiovascular system. In addition, the immunomodulatory properties of statins may provide new indications for these agents in the treatment of autoimmune diseases. PMID- 16035397 TI - Symptomatic treatment of osteoarthritis: paracetamol or NSAIDs? AB - The clinical management of osteoarthritis (OA) is today symptomatic, its main goals being relief of pain and improvement of function. Therapy should be multimodal and composed of non-pharmacological, pharmacological and, if necessary, surgical procedures. Paracetamol and non-steroidal anti-inflammatory drugs (NSAIDs) are evidence-based drugs for the symptomatic relief of OA. Newly published comparative studies have shown that NSAIDs are more effective than paracetamol--in contrast to studies from the early 1990s. Some studies have documented that more severe pain and the presence of inflammation can predict better response from NSAIDs than from paracetamol; on the other hand other studies have not confirmed this. Patient preference studies have shown that patients favour NSAIDs, but up to 40% consider paracetamol at least as effective as NSAIDs. With regard to efficacy, safety and cost, the majority of new guidelines recommend paracetamol as a first-choice analgesic for patients with OA of the knee or hip, and the use of NSAIDs only in cases of inadequate effect of paracetamol and especially in the presence of inflammation. There is much evidence that OA is a phasic disease and it may be that NSAIDs are useful during identifiable periods of inflammatory activity and can be avoided at other times. The concept of the short-term use of NSAIDs during flares and the use of a simple analgesic in the long term seems to be the best variant for the majority of patients with optimal benefit/risk and cost-effectiveness. PMID- 16035398 TI - Effect of nimesulide on the serum levels of hyaluronan and stromelysin-1 in patients with osteoarthritis: a pilot study. AB - This prospective preliminary single-blind study was conducted in patients suffering from osteoarthritis (OA) and requiring non-steroidal anti-inflammatory drugs (NSAIDs) to determine to what extent nimesulide (200 mg/day) and ibuprofen (1200 mg/day) could induce significant changes in the serum levels of matrix metalloproteinase-3 (MMP-3), tissue inhibitor-1 of MMPs (TIMP-1), hyaluronan (HA) and YKL-40 after a therapeutic time period of 28 days. The four biochemical parameters were assessed by using immunoassays. Nimesulide significantly reduced the serum levels of both HA and MMP-3, whereas ibuprofen increased moderately but significantly the serum concentrations of MMP-3 and had no effect on the serum concentrations of HA. The two NSAIDs were unable to change the serum levels of both TIMP-1 and YKL-40. These results suggest that nimesulide might have a favourable effect on the metabolism of OA joints. PMID- 16035399 TI - A benefit/risk assessment of existing therapeutic alternatives for the treatment of painful inflammatory conditions. AB - Pain remains the leading reason for which patients consult their doctors. Pain also motivates over-the-counter sales of analgesic medicines, to be taken orally or even transcutaneously. Prescription medicines usually follow attempts at self medication that fail to achieve the desired results. Acute pain usually subsides spontaneously but medicines are needed until that occurs; in arthritic conditions -especially osteoarthritis--anti-inflammatory drugs work best in short-term administration for flares that aggravate chronic but tolerable pain. In cases of chronic pain that exceeds the level of easy tolerance, anti-inflammatory drugs can reduce the pain to tolerable levels more effectively than simple analgesics and narcotic combinations. The non-steroidal anti-inflammatory drugs (NSAIDs) are among the most useful medicines providing an array of drugs that differ chiefly in time of onset of action, duration of action and persistence in the blood. The benefit they provide is pain amelioration; none is curative. The risks are well known and do not differ greatly among the drugs; unwanted gastrointestinal (GI) effects are the most common, but the skin, kidneys, liver and blood forming organs may also be affected. As the benefits are similar, when balancing risk and benefit it is important to: consider the cause and expected duration of pain, balance the risks (GI unwanted effects far outnumber others), assess the severity and likelihood of specific reactions, and consider the costs--not only of the medicines themselves but also those of treating untoward reactions. Thus NSAIDs number among the most successful therapeutic options of modern medicine and, as pain will continue to require intervention, will likely continue--at least as ancillary medications--even as more definitive treatments are developed. In addition, the target population, its characteristics and the duration and acceptance of the intervention need to be considered. PMID- 16035400 TI - How important is the role of the physician in the correct use of a drug? An observational cohort study in general practice. AB - The correct use of a drug is determined by several important factors. The most significant of these is a correct diagnosis for the choice of the appropriate therapeutic approach, followed by the physician's awareness of each drug's product characteristics--such as indications, contraindications and warnings--and by a careful evaluation of the patient in order to consider possible risk factors, concomitant pathologies and treatments. An adequate knowledge of pharmacological therapy and of the patient's history and disease states could in fact prevent most of the adverse drug reactions attributable to an inappropriate prescription. It is clear that the physician's role in the correct use of a drug is extremely important. Observational studies and surveys evaluating doctors' prescribing habits could be a very useful instrument in identifying medication prescribing errors and the consequent occurrence of adverse events. An open label, multicentre, observational cohort study, recently performed in Ireland and involving more than 9000 patients, investigated the use of non-steroidal anti inflammatory drugs (NSAIDs) in general practice, and in particular the use of diclofenac, nimesulide and ibuprofen. These three drugs were shown to be the NSAIDs most frequently prescribed (80% of total prescriptions) by general practitioners in Ireland. The study was designed and powered to detect differences in the general safety profile of the three NSAIDs and to test prescribing physicians' compliance with each drug's Summary of Product Characteristics (SmPC). The study was performed as closely as possible to the normal prescribing habits of the participating physicians. The three drugs were shown to be well tolerated. In the nimesulide group, the percentage of patients who experienced treatment-related adverse events was lower than that of the diclofenac group and similar to that observed with the use of ibuprofen. A more favourable gastrointestinal safety profile was evident for nimesulide in comparison to diclofenac. The data showed that the study physicians prescribed the drugs with a good degree of compliance (84% of prescriptions) with the information reported in each of the SmPCs. This supported the fact that the adverse events were markedly limited. PMID- 16035401 TI - Religion's evolutionary landscape: counterintuition, commitment, compassion, communion. AB - Religion is not an evolutionary adaptation per se, but a recurring cultural by product of the complex evolutionary landscape that sets cognitive, emotional, and material conditions for ordinary human interactions. Religion exploits only ordinary cognitive processes to passionately display costly devotion to counterintuitive worlds governed by supernatural agents. The conceptual foundations of religion are intuitively given by task-specific panhuman cognitive domains, including folkmechanics, folkbiology, and folkpsychology. Core religious beliefs minimally violate ordinary notions about how the world is, with all of its inescapable problems, thus enabling people to imagine minimally impossible supernatural worlds that solve existential problems, including death and deception. Here the focus is on folkpsychology and agency. A key feature of the supernatural agent concepts common to all religions is the triggering of an "Innate Releasing Mechanism," or "agency detector," whose proper (naturally selected) domain encompasses animate objects relevant to hominid survival--such as predators, protectors, and prey--but which actually extends to moving dots on computer screens, voices in wind, and faces on clouds. Folkpsychology also crucially involves metarepresentation, which makes deception possible and threatens any social order. However, these same metacognitive capacities provide the hope and promise of open-ended solutions through representations of counterfactual supernatural worlds that cannot be logically or empirically verified or falsified. Because religious beliefs cannot be deductively or inductively validated, validation occurs only by ritually addressing the very emotions motivating religion. Cross-cultural experimental evidence encourages these claims. PMID- 16035402 TI - Hallucinations in schizophrenia, sensory impairment, and brain disease: a unifying model. AB - Based on recent insight into the thalamocortical system and its role in perception and conscious experience, a unified pathophysiological framework for hallucinations in neurological and psychiatric conditions is proposed, which integrates previously unrelated neurobiological and psychological findings. Gamma frequency rhythms of discharge activity from thalamic and cortical neurons are facilitated by cholinergic arousal and resonate in networks of thalamocortical circuits, thereby transiently forming assemblies of coherent gamma oscillations under constraints of afferent sensory input and prefrontal attentional mechanisms. If perception is based on synchronisation of intrinsic gamma activity in the thalamocortical system, then sensory input to specific thalamic nuclei may merely play a constraining role. Hallucinations can be regarded as underconstrained perceptions that arise when the impact of sensory input on activation of thalamocortical circuits and synchronisation of thalamocortical gamma activity is reduced. In conditions that are accompanied by hallucinations, factors such as cortical hyperexcitability, cortical attentional mechanisms, hyperarousal, increased noise in specific thalamic nuclei, and random sensory input to specific thalamic nuclei may, to a varying degree, contribute to underconstrained activation of thalamocortical circuits. The reticular thalamic nucleus plays an important role in suppressing random activity of relay cells in specific thalamic nuclei, and its dysfunction may be implicated in the biological vulnerability to hallucinations in schizophrenia. Combined with general activation during cholinergic arousal, this leads to excessive disinhibition in specific thalamic nuclei, which may allow cortical attentional mechanisms to recruit thalamic relay cells into resonant assemblies of gamma oscillations, regardless of their actual sensory input, thereby producing an underconstrained perceptual experience. PMID- 16035403 TI - An evolutionary theory of schizophrenia: cortical connectivity, metarepresentation, and the social brain. AB - Schizophrenia is a worldwide, prevalent disorder with a multifactorial but highly genetic aetiology. A constant prevalence rate in the face of reduced fecundity has caused some to argue that an evolutionary advantage exists in unaffected relatives. Here, I critique this adaptationist approach, and review--and find wanting--Crow's "speciation" hypothesis. In keeping with available biological and psychological evidence, I propose an alternative theory of the origins of this disorder. Schizophrenia is a disorder of the social brain, and it exists as a costly trade-off in the evolution of complex social cognition. Paleoanthropological and comparative primate research suggests that hominids evolved complex cortical interconnectivity (in particular, frontotemporal and frontoparietal circuits) to regulate social cognition and the intellectual demands of group living. I suggest that the ontogenetic mechanism underlying this cerebral adaptation was sequential hypermorphosis and that it rendered the hominid brain vulnerable to genetic and environmental insults. I argue that changes in genes regulating the timing of neurodevelopment occurred prior to the migration of Homo sapiens out of Africa 100,000-150,000 years ago, giving rise to the schizotypal spectrum. While some individuals within this spectrum may have exhibited unusual creativity and iconoclasm, this phenotype was not necessarily adaptive in reproductive terms. However, because the disorder shared a common genetic basis with the evolving circuitry of the social brain, it persisted. Thus schizophrenia emerged as a costly trade-off in the evolution of complex social cognition. PMID- 16035404 TI - Nurses face a lottery over choice of shifts. PMID- 16035405 TI - When health workers get 'wasted'. PMID- 16035406 TI - For the record. PMID- 16035407 TI - How I coped with... an airborne emergency. PMID- 16035408 TI - Introducing matrons' ward rounds to improve care. AB - Modern matrons are expected to act as patient advocates and improve the experience of care. Several initiatives aim to achieve this, including the introduction of matrons' ward rounds. These have proved popular with patients and have identified an increased care need for some vulnerable adults during their hospital stay. PMID- 16035409 TI - Handwashing. PMID- 16035410 TI - Didronel PMO. PMID- 16035411 TI - Developments in child health surveillance programmes. AB - The promotion of good health and well-being for children is essential to improve adult health. Child health surveillance has been crucial in attempting to achieve this goal. The National Service Framework for Children promotes a new child health programme. If it is to be successful, nurses involved in primary care must play a central role. PMID- 16035412 TI - Increasing staff awareness of respiratory rate significance. AB - Respiratory rate is a significant predictor of critical illness and an important part of early-warning scoring. After an audit of patient records revealed a low incidence of respiratory-rate recording, a hospital-wide education programme was set up to raise awareness and improve critical care skills among ward staff. A repeat audit showed significant improvement in recording rates and identified new areas for training. PMID- 16035413 TI - Visiting time preferences of patients, visitors and staff. AB - AIM: To obtain the views of patients, visitors and nursing staff regarding visiting hours. METHOD: Questionnaires, collecting qualitative and quantitative data, were distributed on each of the trust's 35 inpatient adult wards. A total of 863 completed questionnaires were returned - 432 from nursing staff, 227 from patients and 204 from visitors. RESULTS: None of the three groups of respondents preferred completely open visiting; Patients and visitors preferred open visiting with a quiet hour; Nursing staff preferred set visiting hours; One-third of patients were embarrassed about receiving nursing care in front of visitors; One third of patients, visitors and staff did not like visitors to be present at mealtimes. CONCLUSION: Nurses, patients and visitors did not prefer open visiting as first choice, preferring to have a rest period. This highlights the need to consult with service users. PMID- 16035414 TI - Improving the lives of epilepsy patients. PMID- 16035415 TI - Learn about your community in A&E. PMID- 16035416 TI - The NHS is too complex to appreciate. PMID- 16035417 TI - Creating a healthful practice/work environment. PMID- 16035418 TI - Sun and skin: still a burning issue. PMID- 16035419 TI - Assessing malignant melanoma: a case study. AB - Good history and close physical examination with high index of suspicions allowed us to perform an excisional biopsy which was also adequate for the primary treatment of this superficial malignant melanoma. The diagnosis, management, and followup of melanoma have undergone many changes and updates over the past decade, prompting practitioners to keep abreast of updated techniques on newer guidelines in order to offer excellent quality care. PMID- 16035420 TI - Melanoma: treatment guidelines for patients (part 2). AB - The mutual goal of the National Comprehensive Cancer Network (NCCN) and the American Cancer Society partnership is to provide patients and the generalpublic with state of the art cancer treatment information in understandable language. This treatment information is based on the professional guidelines developed by the NCCN. It is intended to assist patients in a discussion with their doctor about the treatment that is best for their specific situation. Part I of these guidelines addressed melanoma risk factors, prevention, diagnosis, stages, prevention, and clinical trials (Vol. 17, No 2, pp. 119-131). Part II describes decision trees for evaluation and treatment. PMID- 16035421 TI - What's your assessment? Prurigo nodularis. PMID- 16035422 TI - Depilatory cream vs. shaving: does it influence recurrences of genital warts? AB - Genital warts are common infections caused by human papilloma viruses (HPV). Although the diagnosis is straight forward and many different treatment choices are present, recurrences are almost inevitable. There are many factors influencing recurrences such as immunity, HPV types, sexual partnership, and hygiene. Hair removal methods may be an issue in recurrence. In this retrospective study, the recurrence rate of genital warts in 50 patients using depilatory creams or shaving as a regular hair removal method before and after treatment by electrocauterization were evaluated. Both methods showed no difference in recurrence rates. Therefore, no depilatory method can be advised as decreasing genital recurrence of genital warts. PMID- 16035423 TI - So, what do I put on this wound? Making sense of the wound dressing puzzle: part II. PMID- 16035424 TI - Atypical fibroxanthoma (AFX). PMID- 16035425 TI - Dissecting cellulitis of the scalp. PMID- 16035426 TI - Patch testing. PMID- 16035427 TI - Don't mess with Texas. PMID- 16035430 TI - Regulatory review: deja vu again! PMID- 16035431 TI - Nappies and the environment: the debate continues. PMID- 16035432 TI - A student's perspective. PMID- 16035433 TI - Midwifery reborn: a metamorphosis. PMID- 16035434 TI - KSF and the midwife. PMID- 16035435 TI - 'I felt like they were all kind of staring at me'. PMID- 16035436 TI - Strengthening midwifery leadership. PMID- 16035437 TI - Student midwives: views of the direct-entry programme. PMID- 16035438 TI - The public health divide. PMID- 16035439 TI - Retention and recruitment of staff. PMID- 16035440 TI - Differences between midwives and nurses. PMID- 16035441 TI - Widening the midwifery network. PMID- 16035442 TI - Every child matters: change for children. PMID- 16035443 TI - Should midwives be promoting and providing home birth? PMID- 16035444 TI - En caul births. PMID- 16035445 TI - Do today's midwifery students spend enough time in practice situations? PMID- 16035446 TI - Best practice for small intestinal biopsy. PMID- 16035447 TI - Complications following full-thickness small intestinal biopsy in 66 dogs: a retrospective study. AB - OBJECTIVES: To retrospectively report the complications seen after full-thickness multiple small intestinal biopsies were performed in 66 dogs. METHODS: Animals that died as a result of enteric wound breakdown were compared with the surviving population to identify fatal risk factors. RESULTS: Seventeen dogs had hypoalbuminaemia and eight had albumin levels below 20 g/I at surgery. Twelve dogs had concurrent disease, of which seven had skin disease. Seven dogs suffered minor complications which resolved with treatment. Eight dogs (12 per cent) died or were euthanased between three and nine days postoperatively (mean [sd] 4.5 [2.1]) due to enteric wound breakdown. Seven of these dogs developed septic peritonitis and one died of haemorrhage. No statistically significant differences were identified in any of the parameters examined for the development of fatal enteric wound dehiscence. CLINICAL SIGNIFICANCE: Full-thickness intestinal biopsy is not a benign procedure. However, this study concludes that there are no consistent predictors for patients at increased risk of enteric wound breakdown. PMID- 16035448 TI - Ultrasonographic evaluation of the thickness of the small intestinal wall in dogs with inflammatory bowel disease. AB - OBJECTIVES: To establish whether the intestinal wall thickness, as measured ultrasonographically, is significantly increased in dogs with inflammatory bowel disease (IBD). The results would provide the information necessary to decide whether measurement of ultrasonographic wall thickness can predict IBD in dogs. METHODS: The intestinal wall thickness of 75 dogs with idiopathic IBD, as measured by ultrasonography, was compared with recently published normal values. IBD was either confirmed histologically (n = 54) or suspected (n = 21). In all cases there was a positive response to immunosuppressive treatment. RESULTS: A positive association between intestinal wall thickness in dogs and either the histological diagnosis or the response to treatment was not found. Ultrasonographic intestinal wall measurements do not appear to be able to establish a diagnosis of intestinal inflammation and may result in a false negative diagnosis in cases of IBD. CLINICAL SIGNIFICANCE: The same 'grey zone' of between 4 and 6 mm used in humans can be used in the canine duodenum to distinguish the normal range, reserving the term 'abnormal' for an intestinal measurement greater than 6 mm in the duodenum and greater than 4.7 mm in the jejunum. PMID- 16035449 TI - Evaluation of elbow incongruency using reconstructed CT in dogs suffering fragmented coronoid process. AB - OBJECTIVES: A retrospective study was undertaken to evaluate elbow joint congruency in dogs suffering fragmented coronoid process (FCP). METHODS: Based on clinical, radiographic and computed tomographic (CT) examinations, elbows were divided into control and FCP groups. Standardised CT reconstructions were formatted in the frontal and sagittal planes. Humeroradial and humeroulnar joint space measurements were obtained from the Images and incongruencies were calculated by comparing the two measurements. RESULTS: Forty-two FCP and 29 control elbows were identified. No incongruencies were noted at the coronoid base. At the level of the coronoid apex, FCP elbows exhibited a significant radioulnar incongruency compared with controls (P < 0.0001), though incongruency was not identified in all cases. Comparing FCP and control elbows at the level of the apex, the humeroradial joint space was increased in FCP elbows (P = 0.0006) whereas no difference was noted in the humeroulnar space. CLINICAL SIGNIFICANCE: This study supports the hypothesis that joint incongruency is associated with FCP in dogs, though is not present in every case at the time of diagnosis. The precise mechanism of development of this incongruency cannot be determined from these data. PMID- 16035450 TI - Hypercalcaemia in two dogs caused by excessive dietary supplementation of vitamin D. AB - A three-year-old Border collie was presented with a two-week history of lethargy, stiff gait, polydipsia and polyuria. Biochemical analysis revealed hypercalcaemia. Serum concentrations of 25-hydroxyvitamin D (25[OH]D) and 1,25 dihydroxyvitamin D (1,25[OH]2D) were markedly elevated and parathyroid hormone was undetectable. Subsequent analysis of the dog's diet revealed that the food contained excessive amounts of vitamin D. The hypercalcaemia resolved following treatment with bisphosphonates and dietary change. Hypervitaminosis D was diagnosed in a second unrelated dog, which had been fed the same brand of dog food as case 1. The dog was also hypercalcaemic and had markedly elevated serum concentrations of 25(OH)D and 1,25(OH)2D. Hypervitaminosis D in dogs has been reported to occur secondarily to ingestion of either rodenticides containing cholecalciferol or antipsoriatic ointments that contain vitamin D analogues. Hypervitaminosis D has also been reported following the treatment of hypoparathyroidism. To the authors' knowledge, this is the first report of hypervitaminosis D in dogs following the accidental over supplementation of a commercial diet with vitamin D. While the benefits of adequate dietary vitamin D are well established in dogs, the potential deleterious effects of over supplementation of vitamin D should also be acknowledged. PMID- 16035451 TI - Hyperammonaemic encephalopathy secondary to selective cobalamin deficiency in a juvenile Border collie. AB - An eight-month-old Border collie was presented with anorexia, cachexia, failure to thrive and stupor. Laboratory tests demonstrated a mild anaemia, neutropenia, proteinuria and hyperammonaemia. Serum bile acid concentrations were normal, but an ammonia tolerance test (ATT) was abnormal. The dog responded to symptomatic therapy for hepatoencephalopathy. When a low serum cobalamin (vitamin B12) concentration and methylmalonic aciduria were noted, the dog was given a supplement of parenteral cobalamin. Two weeks later, a repeat ATT was normal. Cobalamin supplementation was continued every two weeks, and all clinical signs, except for proteinuria, resolved despite withdrawing all therapy for hepatoencephalopathy. A presumptive diagnosis of hereditary selective cobalamin malabsorption was made, based on the young age, Border collie breed, low serum cobalamin concentration and methylmalonic aciduria. Although hereditary selective cobalamin malabsorption in Border collies, giant schnauzers, Australian shepherd dogs and beagles has previously been reported in North America, to the authors' knowledge this is the first report of the condition in the UK and the first to document an abnormal ATT in a cobalamin-deficient dog. PMID- 16035452 TI - Hypocalcaemia associated with low serum vitamin D metabolite concentrations in two dogs with protein-losing enteropathies. AB - Protein-losing enteropathies were diagnosed in two dogs that were initially presented with diarrhoea and weight loss. Plasma biochemistry in both cases revealed low concentrations of albumin, calcium and ionised calcium. Both dogs had an elevated plasma parathyroid hormone concentration and low serum 25 hydroxyvitamin D (25[OH]D) concentration. The first dog was diagnosed with lymphangiectasia on postmortem examination, and the second dog was diagnosed with chronic lymphocytic/ plasmacytic enteritis and severe cystic mucoid changes based on endoscopic duodenal biopsies. While a causal effect was not demonstrated, the protein-losing enteropathies may have caused reduced intestinal absorption of vitamin D leading to low plasma concentrations of ionised calcium and secondary hyperparathyroidism. To the authors' knowledge, this is the first report of low ionised calcium concentrations, low 25(OH)D and 1,25-dihydroxyvitamin D concentrations, and high parathyroid hormone concentrations in dogs with protein losing enteropathies. PMID- 16035453 TI - What is your diagnosis? The principal differential diagnosis for an intraluminal colonic soft tissue structure is intussusception. PMID- 16035454 TI - RCVS Practice Standards Scheme exceeds target. PMID- 16035455 TI - Keeping track of adverse reactions to microchips. PMID- 16035457 TI - Medley of fundraising initiatives at BSAVA Congress. PMID- 16035456 TI - New research into pancreatitis in dogs. PMID- 16035458 TI - World Congress--a fiesta of education and more in Mexico. PMID- 16035459 TI - ADA task force on workforce models. PMID- 16035460 TI - You know them when you see them. PMID- 16035461 TI - Carrying around that DANB pride. PMID- 16035462 TI - The ADAA/DANB Alliance. PMID- 16035463 TI - Being prepared to treat patients with renal disease. PMID- 16035464 TI - Terminology, technology, and troubleshooting for the dental assistant. PMID- 16035465 TI - The good receptionist. PMID- 16035466 TI - Dental Command announces 2005 Soldier & NCO of the Year. PMID- 16035467 TI - A team approach to presenting cosmetic dentistry. PMID- 16035468 TI - Habla Espanol? PMID- 16035469 TI - Responding with a united voice to the Conditions of Coverage! PMID- 16035470 TI - Clinical management of intradialytic hypotension: survey results. AB - Intradialytic hypotension (IDH) is one of the most common complications of hemodialysis treatment. Incidence of IDH varies from 10% to 50% of hemodialysis treatments. Numerous publications address the pathophysiology, effect of frequent episodes, and clinical management of IDH, but no recent publication has studied how IDH is managed in dialysis clinics. Interviews and a mail survey were conducted to develop a better understanding of the clinical issues and concerns surrounding the management of hemodialysis patients who experience episodes of IDH. Results of the survey indicate that IDH is prevalent and that clinical management is protocol--driven and consistent with treatment recommendations in the literature. PMID- 16035471 TI - Preparing for hemodialysis: patient stressors and responses. AB - This qualitative study explored stressors experienced by individuals with chronic kidney disease (CKD). Individuals on hemodialysis were asked to share their recollections regarding stressors they experienced as they approached dialysis. Content analysis was used and the following themes emerged: (a) stressors; (b) emotional responses; (c) learning, preparation, and acceptance; and (d) regret and dissatisfaction. The results of this study have important implications for patient education and support in the care of patients with CKD. PMID- 16035472 TI - Quality of life in patients on dialysis: benefits of maintaining a hemoglobin of 11 to 12 g/dL. AB - Clinical data have consistently demonstrated that improvements in quality of life (QOL) are associated with hemoglobin (Hb) and hematocrit (Hct) levels maintained in the range recommended by the National Kidney Foundation's Kidney Disease Outcomes Quality Initiative (NKF-K/DOQI). Hb levels between 11 and 12 g/dL and Hct levels between 33% to 36% yield significant improvements in a wide variety of parameters in the physical, cognitive, and psychological/social domains. PMID- 16035473 TI - Saving a brachiocephalic fistula using lipectomy. AB - Many studies have demonstrated that functioning AVFs provide better vascular access outcomes compared to prosthetic devices. In April 2004, the Centers for Medicare and Medicaid Services (CMS) launched a "safe vascular access through collaborative fistula first initiative" (CMS, 2004). This breakthrough proposal cites studies that show increased mortality in patients who do not dialyze with a fistula. Veins too deep for needle cannulation are usually free of scars from intravenous (IV) sticks and, thus, are excellent options for fistula creation. Lipectomy can remove barriers, such as vein depth and limb obesity, creating an opportunity to have a functioning fistula. PMID- 16035474 TI - ACE inhibitors and ARBs: antihypertensive medications in CKD. PMID- 16035475 TI - K/DOQI gets to the heart of managing dyslipidemias in patients with CKD. AB - There is a wealth of data in the general population regarding interventions to reduce cardiovascular risk. Unfortunately, most of these studies exclude patients with chronic kidney disease. As a result, the lack of CKD specific data has resulted in a lack of attention and intervention. With the epidemic levels of cardiovascular disease in patients on dialysis, the NKF has established these K/DOQI guidelines in an effort to get to the "heart" of dyslipidemias and ultimately to assist the health care team in their effort to improve CKD patient outcomes. In addition, the National Kidney Foundation currently has draft K/DOQI Clinical Practice Guidelines for Cardiovascular Disease in the public review process. These new guidelines will elaborate on areas not covered in the dyslipidemia guidelines. PMID- 16035476 TI - Home vs. in-center dialysis: reassessing the burdens. PMID- 16035477 TI - Discussion of patient recruitment and the informed consent process in clinical drug trials. PMID- 16035478 TI - [Sex differences in cardiology, pharmacology and immunology. Women respond differently]. PMID- 16035479 TI - [Sex specific drug therapy. Women have more trouble with side effects]. PMID- 16035480 TI - [Immunologic differences between the sexes. Women react more aggressively]. PMID- 16035481 TI - [Backache in gardening. ECG clarifies the "BWS syndrome"]. PMID- 16035482 TI - [More than 100 virus infections caused by needle stick injuries. How safe is performance in your practice? (interview by Dr. Thomas Meissner)]. PMID- 16035483 TI - [Minimal invasive procedures for back pain are controversial. Which methods help whom]. PMID- 16035484 TI - [The summer is also associated with risks and side effects: how you can help your patients survive the heat]. PMID- 16035485 TI - [Sports and leisure injuries in summer]. AB - In the first days of summer, there is an increase in the number of summer-related accidents and injuries. Typical for the types of sports practiced in summer, such as ball games, cycling or skating are injuries to the wrist and ankle, knee, head and shoulder. The most frequent victims of swimming accidents are children under four years of age, and adolescents aged between 15 and 19 years. During grilling, burns often occur, most of which, however, are superficial and can be treated in the doctor's office. The incidence of dog bites also increases in summer. In such cases consideration must be given not only to tetanus boosters, but also to the possibility of an infection with rabies. PMID- 16035486 TI - [Heat-related problems in the elderly]. AB - In particular in the elderly patient, exposure to heat can lead to disturbances of the circulatory system and of the water and electrolyte balance. Provided that certain prophylactic measures are taken, serious problems are unlikely to occur. Food and drink should be matched to the ambient temperature, and permanent medication should be checked. In the case of confused persons, nursing personnel should substitute for any failure to make the necessary acclimatization changes (appropriate clothing). In the case of incontinent patients, it must be remembered that the diapered area is not available for radiating off heat. PMID- 16035487 TI - [Reactions of the skin to solar radiation]. AB - When spending time out of doors in summer, certain rules of behavior need to be observed. Environmental factors such as, for example, water, the beach and the midday sun all have a major influence on the UV radiation burden to which people may be subjected. In addition, the sun's rays can interact with certain medications, alcohol and drugs, as also cosmetics, and trigger pathological skin reactions. A permanently sun-damaged skin is not only very difficult to treat, but is also frequently associated with malignant changes. Moderate exposure to the sun after prior gradual acclimatization is to be recommended. Patients with increased photosensitivity require special photoprotective agents. PMID- 16035488 TI - [Insect stings and their sequelae]. AB - In the summer months, insect stings are a common reason for seeking help from the general physician. In the majority of cases a local reaction is seen, but, far less often, an anaphylactic reaction may also occur. Such an acute situation requires calm but decisive action, and the initiation of an evaluation by an allergy specialist. The family doctor should regularly review the facilities for treating an anaphylactic reaction in his office, and, where necessary, optimize them. PMID- 16035489 TI - [Structured educational programs for geriatric patients with diabetes mellitus]. AB - Two-thirds of all diabetics are older than 60 years. Owing to the presence of multimorbidity and functional impairments, one-half of these diabetics must be considered "geriatric patients". This category of patients in particular requires a special educational program aimed at improving their situation and quality of life on the basis of good diabetic control. Currently, two educational concepts aimed in particular at elderly patients with type II diabetics and cognitive deficits are available and are described in the present paper. Education for diabetics with impairment of cognitive performance is available to such diabetics irrespective of age. The educational program: "Structured Education for Persons with Type II Diabetes in Old Age", newly developed by the Working Group Diabetes and Geriatrics of the German Diabetes Society teaches self-help skills, offers opportunities for empowerment, and has proved successful also in geriatric patients with mild cognitive deficits. PMID- 16035490 TI - [Surprising outcome. Quality management engenders losses]. PMID- 16035491 TI - [Bavarian colleagues prevail. General practitioner contract forced by DMP strike]. PMID- 16035492 TI - [20-30% of the population is afflicted. Gastitis is a continous burn in general practice]. PMID- 16035493 TI - [Functional dyspesia or reflux disease. Therapy with PPI often a good approach]. PMID- 16035494 TI - [Etiology and therapy of halitosis. From abscess to coated tongue]. PMID- 16035495 TI - [Diagnosis at a glance. Major crawling symptom]. PMID- 16035496 TI - Key developments in paediatrics. PMID- 16035497 TI - Managing constipation and soiling. PMID- 16035498 TI - Type 1 diabetes in children: a review. PMID- 16035499 TI - Attention deficit hyperactivity disorder. PMID- 16035501 TI - Appearance of the oocyte activation of mouse round spermatids cultured in vitro. AB - This study was undertaken to determine the expression time of fertilization and oocytes activation abilities of spermatids in the mouse. When elongating or elongated spermatids isolated from fresh testes of adult males (B6D2F1) were injected into mature mouse oocytes, both spermatids could activate the mature oocytes and occur fertilization. On the one hand, the round spermatids could not activate mature oocytes, when microinjected into oocytes. In some experiments, recovered round spermatids were cultured under co-culture systems using Sertoli cells as a feeder cell. Under the co-culture system, developed elongating spermatids could stimulate and fertilized mature oocytes. These results indicate that the start of oocyte activation appearance is between the stage of round spermatid and elongating spermatids and the activation ability increases with the advance of spermiogensis. On the other hand, round spermatids isolated from males of ICR strain mouse already have the oocyte activation ability and the fertilizing ability. The result obtained suggests that the expression time of the oocyte activating ability is difficult between the mouse strain. PMID- 16035502 TI - Human sperm centrosomal function during fertilization, a novel assessment for male sterility. AB - In human fertilization, the sperm introduces the centrosome-the microtubule organizing center-and microtubules are organized within the inseminated egg from the sperm centrosome. These microtubules form a radial array, the sperm aster, the functioning of which is essential for pronuclear movement for the union of the male and female genomes. We established functional assay for human sperm centrosomal function, by using heterologus ICSI system with bovine and rabbit eggs. After human sperm incorporation into mammalian egg, we observed that the sperm aster was organized from sperm centrosome, and the sperm aster enlarged as the sperm nuclei underwent pronuclear formation. The normal human sperm aster formation rate at 6 h post-ICSI were 60.0% in bovine egg and 36.1% in rabbit egg, respectively. However, sperm aster formation rate following heterologus ICSI into bovine eggs with teratozoospermia (globozoospermia, dysplasia of fibrous sheath) were low. These data indicate that human sperm centrosomal function is low in abnormal shaped sperm. Wherus, elucidation of human sperm centrosomal function can lead us to find a new type of failure in "post ICSI events in fertilization". PMID- 16035503 TI - Complete fertilization failure in ICSI. AB - Fertilization failure is one of the causes of infertility that becomes evident only after in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) have been attempted. Although the frequency of incidence of fertilization failure is low, if fertilization failure is encountered, medical treatment is usually stopped and serious psychological damage may occur to the patient. While fertilization failure in IVF can be dealt with using ICSI, there is no treatment for fertilization failure in ICSI. At present, clinical investigations are being conducted to evaluate oocyte activation in combination with ICSI to cope with fertilization failure of ICSI. PMID- 16035504 TI - Oocyte mitochondria: strategies to improve embryogenesis. AB - Mitochondria play a central role to provide ATP for fertilization and preimplantation embryo development in the ooplasm. The mitochondrial dysfunction of oocyte has been proposed as one of the causes of high levels of developmental retardation and arrest that occur in preimplantation embryos generated using Assisted Reproductive Technology. Cytoplasmic transfer (CT) from a donor to a recipient oocyte has been applied to infertility due to dysfunctional ooplasm, with resulting pregnancies and births. However, neither the efficacy nor safety of this procedure has been appropriately investigated. In order to improve embryogenesis, we observed the mitochondrial distribution in ooplasma under the several conditions using mitochondrial GFP-transgenic mice (mtGFP-tg mice) in which the mitochondria are visualized by GFP. In this report, we will present our research about the mitochondrial distribution in ooplasm during early embryogenesis and the fate of injected donor mitochondria after CT using mtGFP-tg mice. The mitochondria in ooplasm from the germinal vesicle stage to the morula stage were accumulated in the perinuclear region. The mitochondria of the mtGFP tg mouse oocyte transferred into the wild type mouse embryo could be observed until the blastocysts stage, suggesting that the mtGFP-tg mice oocyte is very useful for visual observation of the mitochondrial distribution in the oocyte, and that the aberrant early developmental competences due to the oocyte mitochondrial dysfunction may be overcome by transferring the "normal" mitochondria. PMID- 16035505 TI - Establishment and characterization of a human ovarian small cell carcinoma, hypercalcemic type, cell line (OS-1) secreting PTH, PthrP and ACTH--special reference to the susceptibility of anti-cancer drugs. AB - We successfully established a novel cell line (OS-1) derived from human ovarian small cell carcinoma, hypercalcemic type secreted PTH, PTH-rP and ACTH. The OS-1 cell line was established from metastatic focus of uterus. A patient was 25-year old Japanese woman. The first she received left ovariectomy on April 2002. The histopathological diagnosis was ovarian small cell carcinoma, pT2c, Nx, Mx. Then on June 2003, metastatic focus of uterus was ectomied. A part of the recurrent tumor of uterus was cut into small pieces with razor blades, and dissociated with 0.1% trypsin-0.02% EDTA/ PBS(-) solution at room temperature. The single cells and small cell clusters were seeded into 60mm dishes and cultured in growth medium (GM: DMEM/F12 supplemented with 20% fetal bovine serum and 0.1% non essential amino acids solution) at 37 degrees C, 4.7% CO2 in humidified air. Medium was exchanged twice a week. The OS-1 cells grew as floating cultures in the dishes. Radioimmunoassay of the conditioned media was revealed that the cultures secreted large amount of PTH, PTHrP and ACTH simultaneously. Susceptibilities of anti-cancer drugs to the OS-1 cells were examined using oxygen electrode meter (Daikin), and the results suggested VLB and TXL were effective, and CDDP, CPT-11, VP-16, VCR, CPA, MMC and CBDCA were not effective. In our knowledge, it is the first report that the cell line secreting PTH, PTHrP and ACTH was successfully established from ovarian small cell carcinoma, hypercalcemic type. We expect that OS-1 cell line contribute to study on the mechanism of ectopic hormone secretion and susceptibility of anti cancer drugs to the small cell carcinoma. PMID- 16035506 TI - In vitro culture of various typed meningiomas and characterization of a human malignant meningioma cell line (HKBMM). AB - We placed on culture the 13 cases of meningiomas, succeeded in making a primary culture of 10 cases and maintained 5 cases in vitro over considerable period of time (over three month), and one cell line derived from a malignant meningioma were established. In the early period of the primary culture, meningioma cells were spindle- or round-shaped cells. In the case of psammomatous type, the cultured cells were characterized as forming psammoma bodies. A cell line designated "HKBMM" was established from a human malignant meningioma occurred from frontal lobe. This line grew well without interruption for 5 years and was subcultivated over 120 times. The cells were spindle and fibrous in shape, and neoplastic and pleomorphic features, and multilayering without contact inhibition. The cells proliferated rapidly, and the population doubling time was about 29 hours. The chromosome number showed a wide distribution of aneuploidy. The mode was in the diploid range. The culture cells were easily transplanted into the subcutis of nude mice and produced the tumor resembling the original tumor. PMID- 16035507 TI - Implementing research. PMID- 16035508 TI - Nurses be proud. PMID- 16035509 TI - About the living will. PMID- 16035510 TI - Life is precious. PMID- 16035511 TI - Mixed-gender wards. PMID- 16035512 TI - A moment of reflection. PMID- 16035513 TI - The Walkerton Health Study. AB - The contamination of the Walkerton, Ontario, municipal water with E. coli O157:H7 and Campylobacter in May 2000 resulted in at least 2,300 cases of gastrointestinal illness. There were 28 confirmed cases of hemolytic uremic syndrome, the most severe kidney complication. The provincial Ministry of Health and Long-Term Care determined that a study needed to be conducted on the long term health effects associated with drinking the contaminated water. The Walkerton Health Study, a seven-year project, was established as a screening and treatment clinic to identify and treat those people experiencing illness and to study long-term health effects. This article describes the challenges, infrastructure support, staffing, and recruitment and retention efforts required to screen over 4,000 people in a yearly clinic visit. Clinical and laboratory algorithms are used to identify participants requiring specialist assessment. Design of the computer-based survey includes advanced data entry and display control, essential to ensuring accurate data for analysis. Findings from Years 1, 2 and 3 are briefly discussed. PMID- 16035514 TI - Global health and equity. Part 1: setting the context. AB - Equity is considered one of the prerequisites and conditions for health. The pursuit of global health and equity is a relevant goal for nursing, based on the belief that health is a core phenomenon of interest to the discipline. In this article, the first in a two-part series, global health, equity and globalization are analysed. In addition, factors that contribute to global health inequity, including their relevance for nurses and nursing, are examined. PMID- 16035515 TI - Choosing the right graduate program. PMID- 16035517 TI - Bullying in the workplace. PMID- 16035516 TI - Ethics in action. PMID- 16035518 TI - Nurses. Worth listening to. PMID- 16035519 TI - And it all came tumbling down...new industrial relations legislation to hurt nurses. AB - When the Howard Government gains total control of Parliament in July this year, it has announced it will introduce legislation which will dismantle Australia's industrial relations system. The Australian Nursing Federation (ANF) and the Queensland Nurses' Union (QNU) expect the new legislation will reduce the role of unions, put downward pressure on wages and conditions and dilute the power of the Australian Industrial Relations Commission (AIRC) to hear and resolve disputes. The changes will not only affect nurses' wages and conditions, but threaten to undermine the profession itself. PMID- 16035520 TI - The NZ experience--a cautionary tale. AB - New Zealand nurses have already faced the changes awaiting Australian nurses and are still recovering from more than a decade of industrial reforms which left their wages far below other professions, eliminated many of their employment rights, and forced many NZ nurses to move overseas to find work. The latest national employment agreement promises to begin to rectify the damage, but as New Zealand Nurses Organisation (NZNO) president Jane O'Malley told Fiona Armstrong, it's a long way back from the destruction created by the 1991 Employment Contract Act. PMID- 16035521 TI - Bundaberg Hospital Inquiry could spread further afield. PMID- 16035522 TI - Birthing services under spotlight in Queensland. PMID- 16035523 TI - Who is your new editor? PMID- 16035524 TI - Court decision means more claim rejections for Michigan dentists. PMID- 16035525 TI - She says she's pregnant--do I still have to hire her? PMID- 16035526 TI - Big budget surplus for ADA reported. PMID- 16035527 TI - Planning for an IRS audit. PMID- 16035528 TI - Real answers for your collections problems. PMID- 16035529 TI - The donated dental services program: why $5 million in donated care still isn't enough. PMID- 16035530 TI - How to determine the real value of your practice. PMID- 16035531 TI - Professional tolerance. PMID- 16035532 TI - Restructuring the Hawaii Dental Association. PMID- 16035533 TI - Does outsourcing enhance productivity? PMID- 16035534 TI - The virtual accounting department: advantages of outsourcing payroll and accounting functions. PMID- 16035535 TI - New practice management software feature makes increasing case acceptance even easier. PMID- 16035536 TI - Leasing employees: is it the right choice for you? PMID- 16035537 TI - Modesta "Joji" Gaerlan-Tokunaga: being a dentist is a dream come true. PMID- 16035538 TI - The Masunaga dental clan from Wahiawa. PMID- 16035539 TI - New workers' compensation medical fee schedule. PMID- 16035540 TI - Building strong families. PMID- 16035541 TI - CVV in women with HIV. PMID- 16035542 TI - Circumcision is unjustifiable. PMID- 16035543 TI - Kentucky presses on for CS privileges. PMID- 16035544 TI - Saving time with group care. PMID- 16035545 TI - Stabilization after eye trauma. PMID- 16035546 TI - Pain, inflammation and a nodule after IV medication. PMID- 16035547 TI - Patient information. Caring for cuts and burns at home. PMID- 16035548 TI - Handle with care. An overview of burn injury. PMID- 16035549 TI - Chronic venous insufficiency. Preventing leg ulcers is primary goal. PMID- 16035550 TI - Creating the optimal environment. An overview of dressings for chronic wounds. PMID- 16035551 TI - Intrauterine devices. A modern alternative to female sterilization. PMID- 16035552 TI - The best kept secret. Pelvic floor muscle therapy for urinary incontinence. PMID- 16035553 TI - HIV aware in primary care. A refresher on assessment and treatment. PMID- 16035554 TI - Stress fractures of the pelvis and lower extremities. Diagnosis and management. PMID- 16035555 TI - Firearm safety in the home. The role of health care providers. PMID- 16035556 TI - The power of pets. PMID- 16035557 TI - Respiratory medicine in general practice: what is the role of the pulmonary specialist? PMID- 16035558 TI - Underuse of spirometry by general practitioners for the diagnosis of COPD in Italy. AB - BACKGROUND: The Global Initiative for Chronic Obstructive Lung Disease (GOLD) underlines that spirometry is the gold standard as the most reproducible, standardised, and objective way of measuring airflow limitation in the diagnosis and assessment of Chronic Obstructive Pulmonary Disease (COPD). However, studies undertaken in different countries have suggested a widespread underuse of spirometry by general practitioners to establish the diagnosis of COPD. Precise estimates of the prevalence of physician-diagnosed COPD in Italy are not currently available. In collaboration with the Italian Academy of General practitioners (SIMG) we have investigated the degree of use of spirometry to establish the diagnosis of COPD in Italy. METHODS: A standardised questionnaire has been self-administered to a sample of 2425 Italian general practitioners (representing 5% of all the Italian doctors involved in general practice). They have been chosen to cover each of the Italian counties. RESULTS: The prevalence of physician-diagnosed COPD was found to be approximately 4%. However, 30% of general practitioners do not use spirometry to establish the diagnosis of COPD. The main reasons given for the failure to use spirometry are (i) that spirometry is not necessary for the diagnosis of COPD or (ii) there are logistical limitations to the access of the patients to lung function laboratories. CONCLUSIONS: This data suggests that contrary to GOLD Guidelines, in Italy, as with other countries, spirometry is not always used in the diagnosis of COPD. There is a clear necessity for further education initiatives targeted to this group of physicians. PMID- 16035559 TI - Nutritional status and airflow obstruction: two independent contributors to CO diffusing capacity impairment in COPD. AB - BACKGROUND: The association between weight loss and Chronic Obstructive Pulmonary Disease (COPD) has been recognised from many years. Based on the evidence that nutritional status reflects metabolic disturbances in COPD, the relationship between body mass index (BMI), severity of airflow obstruction and CO diffusing capacity (DL(CO)), that is the functional hallmark of emphysema, is relevant to the management of COPD phenotypes. METHODS: We reviewed 104 patients with COPD (82 males), aged 66 +/- 9 years (mean +/- SD). Height averaged 165 +/- 8 cm, weight 71 +/- 16 Kg, FEV1 50 +/- 18 (% of predicted), RV 169 +/- 49%, and DL(CO) 56 +/- 26%. Multiple linear regression was performed using BMI, FEV1 and RV, as explanatory variables for DL(CO). Patients were also classified into four groups according to BMI < or = 18.5 (low), > 18.5 and < or = 25 (ideal), > 25 and < or = 30 (overweight), > 30 (obese), and post-bronchodilator FEV1 < 50%. Using this categorisation, a two-factor analysis of variance, testing for interaction and main effects (BMI and FEV1) was performed as confirmatory analysis for the association between BMI (kg/m2), FEV1% and DL(CO)%. RESULTS: FEV1 and BMI were significantly and independently associated to DL(CO) according to the equation: DL(CO) = -18.32 + 0.65 x FEV1 + 1.59 x BMI (R2 = 0.40, p < 0.0001). The contribution of RV % to DL(CO) % was largely non-significant (p = 0.16). A close relationship was found between BMI (kg/m2) and DL(CO) %, for all of the four BMI groups segregated by post-bronchodilator FEV1 %, (p < .0001). No interaction was found between these two factors (p = 0.30). CONCLUSION: Nutritional status as assessed by BMI contributes substantially to impairment of DL(CO) independently of the severity of airflow obstruction. This data confirms the association between emphysematous process and weight loss in advanced COPD, independent of the airflow obstruction severity. PMID- 16035560 TI - CT in differential diagnosis of benign and malignant pleural disease. AB - BACKGROUND: CT plays a valuable role in assessment of patients with a wide variety of diseases of the pleura, and pulmonologists should be aware of the significance of different CT findings for the differential diagnosis of benign and malignant pleural diseases. METHODS: 155 patients with pleural disease who had undergone CT scans of the lungs and thorax before treatment were enrolled. We retrospectively reviewed CT findings in 146 patients with proven pleural disease. RESULTS: Fifty-nine of the cases were malignant, 87 of them had benign pleural diseases. CT findings that were helpful in distinguishing malignant from benign pleural disease were: 1) pleural nodularity; 2) rind; 3) mediastinal pleural involvement; and 4) pleural thickening greater than 1 cm. The sensitivities and specificities were 37%/97%, 22%/97%, 31%/85%, 35%/87%, respectively. CT findings differentiating malignant pleural mesothelioma from metastatic pleural disease were identified. Findings for malignant mesothelioma were as follows: 1) involvement of interlobar fissure (sensitivity 30%, specificity 92%), 2) pleural thickening greater than 1 cm (sensitivity 60%, specificity 77%). Whereas, findings for metastatic pleural disease were mediastinal/hilar lymph node enlargement and lung parenchymal involvement (P < .05). CONCLUSION: CT is helpful in the differential diagnosis of pleural diseases, particularly in differentiating malignant from benign conditions and metastatic pleural disease from malignant mesothelioma. PMID- 16035561 TI - GP management of community-acquired pneumonia in Italy: the ISOCAP study. AB - BACKGROUND: Community-Acquired Pneumonia (CAP) is still a significant problem in terms of incidence, mortality rate, particularly in infants and the elderly, and socioeconomic burden. General Practitioners (GPs) are the first reference for patients with this disease, but there are few published studies regarding the outpatient treatment of CAP. METHODS: The ISOCAP study aimed to identify the type and outcome of the diagnostic-therapeutic management of CAP by GPs in Italy, within the framework of developing a closer interrelationship between GPs and pulmonary specialists. Thirty-six Pulmonary Divisions throughout Italy each contacted 5 local GPs who agreed to recruit the first 5 consecutive patients who consulted them for suspected CAP within the study's 1-year observation period. RESULTS: A total of 183 GPs took part in the study and enrolled, by the end of the observation period, 763 CAP patients; of these, complete data was available for 737 patients [males=373, females=364, mean age (+/- SD) 58.8 +/- 19.6 years]. 64.4% of patients had concomitant diseases, mainly systemic arterial hypertension and COPD. Diagnosis of CAP was based by GPs on physical examination only in 41.6% of cases; in the remaining chest X-ray was also performed. In only 4.6% of patients were samples sent for microbiological analysis. All patients were treated with antibiotics: 76.7% in mono-therapy, 23.3% with a combination of antibiotics. The antibiotic class most prevalently used in mono-therapy was cephalosporin, primarily ceftriaxone; the most frequently used combinations were cephalosporin + macrolide and cephalosporin + quinolone. Mono-therapy was effective in 70% of cases, the combination of two or more antibiotics in 91.2% of patients. Overall treatment efficacy was 94.7%; hospitalisation was required in 8.5% of cases. CONCLUSIONS: Outpatient management of CAP by GPs in Italy is effective, hospitalisation being necessary only in the most severe cases due to age, co-morbidities or extent of pneumonia. This signifies a very significant savings in national health costs. PMID- 16035562 TI - A simple and easy home-based pulmonary rehabilitation programme for patients with chronic lung diseases. AB - BACKGROUND AND AIM: To develop a simple and easy home-based pulmonary rehabilitation programme and investigate its effectiveness. METHODS: Patients with stable chronic lung disease were divided into a rehabilitation group (n = 25) and a control group (n = 18). Rehabilitation consisted of education and 12 weeks of enforced aerobic and muscle-strengthening exercises. Aerobic exercise training was performed mostly by walking based on the functional capacity of the patients assessed by the maximal incremental exercise test. Patients visited hospital every two weeks for evaluation and a new exercise regimen. RESULTS: Five patients dropped out of the rehabilitation group but three were due to development of unrelated diseases, five controls did not co-operate in the second evaluation. After 12 weeks of rehabilitation, exercise capacity (maximum work load and VO2max), exercise endurance, 6-minute walking distance, and quality of life measured by the St. George Respiratory Questionnaire had significantly improved in the rehabilitation group but not in the controls. At a follow-up evaluation one year after the rehabilitation, some exercise parameters were still significantly higher than baseline in the rehabilitation group. CONCLUSIONS: We developed a simple home-based pulmonary rehabilitation programme, which seems to be clinically feasible and effective. PMID- 16035563 TI - Risk factors for tuberculosis. AB - The risk of developing tuberculosis is dependent on both the risk of being infected and the risk of infection leading on to active disease. The former will depend on the incidence of tuberculosis in the community where the individual lives or works. The latter will depend on many factors impinging on the individual both genetic and environmental. The greatest single risk factor for developing tuberculosis from infection is concurrent HIV infection. Where these two infections are prevalent tuberculosis case rates have risen dramatically and will continue to do so unless either infection can be curtailed. PMID- 16035564 TI - Complementary and alternative medicine for the treatment and diagnosis of asthma and allergic diseases. AB - The use of Complementary/Alternative Medicines (CAM) is largely diffused and constantly increasing, especially in the field of allergic diseases and asthma. Homeopathy, acupuncture and phytotherapy are the most frequently utilised treatments, whereas complementary diagnostic techniques are mainly used in the field of food allergy-intolerance. Looking at the literature, the majority of clinical trials with CAMS are of low methodological quality, thus difficult to interpret. There are very few studies performed in a rigorously controlled fashion, and those studies provided inconclusive results. In asthma, none of the CAM have thus far been proved more effective than placebo or equally effective as standard treatments. Some herbal products, containing active principles, have displayed some clinical effect, but the herbal remedies are usually not standardised and not quantified, thus carry the risk of toxic effects or interactions. None of the alternative diagnostic techniques (electrodermal testing, kinesiology, leukocytotoxic test, iridology, hair analysis) have been proved able to distinguish between healthy and allergic subjects or to diagnose sensitizations. Therefore these tests must not be used, since they can lead to delayed or incorrect diagnosis and therapy. PMID- 16035565 TI - Bronchiolitis obliterans organising pneumonia (BOOP) in a lung cancer patient after lobectomy. AB - A 79 year-old patient with lung cancer underwent a standard thoracotomy and lobectomy. Postoperatively, he developed low-grade fever and dyspnoea. Chest X rays showed progressive lung infiltrates, which was subsequently diagnosed to be Bronchiolitis Obliterans Organizing Pneumonia (BOOP) by transbronchial lung biopsy. He responded well to corticosteroid therapy. The case report is followed by a brief discussion on BOOP in association with lung cancer and thoracotomy. PMID- 16035566 TI - Pulmonary alveolar microlithiasis: CT and pathologic findings in 10 patients. AB - BACKGROUND AND AIM: To evaluate CT findings of pulmonary alveolar microlithiasis and correlate the CT with the pathologic findings. METHODS: The study included 10 patients with pathologically proven microlithiasis. Two independent observers evaluated the presence, extent and distribution of the CT findings. CT findings were compared with those at autopsy in two patients and with transbronchial biopsy in eight patients. RESULTS: All patients had a myriad of calcified nodules measuring approximately 1 mm in diameter. Close apposition of the nodules resulted in areas of ground-glass attenuation and consolidation, which were the predominant abnormality on CT in all 10 patients, involving 41% +/- 16.3 (mean +/ SD) and 30% +/- 4.8 of the lung parenchyma, respectively. Calcifications were also seen along interlobular septa, bronchovascular bundles and pleura. Other findings included interlobular septal thickening, thickening of bronchovascular bundles, nodules, and subpleural cysts. There was a solid agreement between the observers for the presence (kappa value; 0.77) and extent (Spearman rank correlation; r = 0.81 to 1.0 p < 0.01) of abnormalities. Autopsy specimens demonstrated microliths in alveolar airspaces and along interlobular septa, bronchovascular bundles and pleura. Subpleural small cysts were shown to represent dilated alveolar ducts. CONCLUSION: Pulmonary microlithiasis is characterised by the presence of numerous small, calcified nodules, calcifications along interlobular septa, bronchovascular bundles and pleura, ground-glass opacities, consolidation, and subpleural cysts. The cysts represent dilated alveolar ducts. PMID- 16035567 TI - Value of nodal drainage patterns and tumor location from lymphoscintigraphic mapping in detecting axillary sentinel lymph node status in breast cancer: experience at Kaohsiung Medical University Hospital. AB - Sentinel lymph node (SLN) biopsy is an alternative to axillary node dissection for staging breast cancer treatment. In this article, we investigated nodal drainage patterns and tumor location using lymphoscintigraphy to predict the axillary sentinel lymph node status. We enrolled 88 patients with clinically suspicious or biopsy-proven breast cancer from March 2001 to October 2002. The average age of subjects was 48 +/- 4 years and the clinical stage was T(1-2)N0Mx. Tc-99m sulfur colloid was used in a hybrid combination of subdermal and perilesional injections around the selected comers of the tumor or biopsy site. Sentinel lymphoscintigraphy was performed 16-20 hours before surgery. Sentinel nodes were marked on the skin. An intraoperative gamma probe was used to confirm the sentinel lymph node location before biopsy. Most primary tumors were in the outer upper quadrant (52.3%), followed by the inner upper quadrant (17.0%), outer inferior quadrant (12.5%), central areolar area (11.4%), and inner inferior quadrant (6.8%). The nodal drainage patterns on 2-hour lymphoscintigraphy were as follows: axillary alone (76.1%), internal mammary nodes alone (1.1%), both axillary and internal mammary nodes (11.4%), and no drainage (11.4%). Internal mammary lymphatic drainage is related to tumor location in the inner quadrants of the breast. About 11.4% of all patients had poorly identified SLNs on lymphoscintigraphy within a 2-hour period, but there was improvement in the overall detection rate up to 95% by intraoperative gamma probe the next day. Preoperative lymphoscintigraphic mapping has value in providing individual lymphatic drainage patterns and tumor location that are important in the interpretation of the results of SLN biopsy during surgery. PMID- 16035568 TI - Coincidence planar imaging for dynamic [18F]FDG uptake in nude mice with tumors and inflammation: correlated with histopathology and micro-autoradiography. AB - The Institute of Nuclear Energy Research of Taiwan has developed a dynamic coincidence detection device for positron emitted radiotracer pharmacodynamic study in small mice models. In this study, we set up an experimental paradigm by determining [fluorine-18]-2-deoxy-2-fluoro-D-glucose ([18F]FDG) dynamic uptake in tumors and inflammations in nude mice as the foundation for future applications in therapy development. Histopathology and micro-autoradiography of these tumors and inflammations were obtained for confirmation. Dynamic coincidence planar images of six tumors and two inflammations in nude mice were acquired over 4 hours immediately after injection of 25.9 MBq of [18F]FDG into the right thigh of each animal. After image reconstruction, the lesion-to-background ratios were calculated in regions of interest over the lesion and contralateral thigh to determine the equilibrium status of the radiotracer. All mice were sacrificed for histopathologic examination and six of the mice were examined with micro autoradiography. [18F]FDG uptake in tumors and inflammations both reached equilibrium about 3 hours after injection. At equilibrium, [18F]FDG uptake into tumors was two to four times higher than the background. Uptake into the 4-day and 8-day inflammations was 2.3 and 5.5 times higher than the background, respectively. Histopathology showed macrophage and neutrophil infiltration around the tumors and in the inflammations. Micro-autoradiography showed dense silver grains in the granulation tissue surrounding the tumors and inflammations. The preliminary results suggested that dynamic [18F]FDG coincidence planar imaging can help in determining the suitable time for static [18F]FDG imaging in nude mice models. The optimal time for static [18F]FDG positron emission tomography imaging was around 3 hours after injection. The paradigm for determining a dynamic [18F]FDG uptake pattern was demonstrated for future new therapeutic drug experimental use. PMID- 16035569 TI - Cognitive dysfunction after acute lacunar infarct. AB - Vascular dementia and vascular cognitive impairment have attracted more attention recently due to their association with increased risk of death and institutionalization. The purpose of the present study was to detect and identify the characteristics of cognitive impairments during the early stage of lacunar stroke. The subjects consisted of 23 consecutive first-ever acute lacunar infarction patients who were admitted to the Department of Neurology, Kaohsiung Municipal Hsiao-Kang Hospital, Taiwan, from November 2001 to October 2002. The National Institutes of Health Stroke Scale and Cognitive Abilities Screening Instrument (CASI) were used to evaluate stroke severity and cognitive function, and assessments were performed by a neurologist and psychologist, within 10 days of stroke onset. Of the 23 patients, 21 (91.3%) had CASI scores below their respective cutoff values and all patients had cognitive impairment in at least one cognitive domain in CASI. There were no significant correlations between CASI abnormality (below the cutoff value) and patient age, education, or the interval from stroke onset. Recent memory impairment was the most often impaired cognitive domain on CASI (19 patients, 82.6%). There were significant correlations between recent memory and "attention or concentration"(correlation coefficient, 0.52; p < 0.05), and "abstraction and judgment" (correlation coefficient, 0.44; p < 0.05). The correlations between recent memory and other domains were not significant. It was concluded that cognitive impairment after acute lacunar infarct is quite common and recent memory is the most often impaired cognitive domain. This may have been caused by the location of the specific lesion as well as by the impairment in "attention or concentration" or "abstraction and judgment". PMID- 16035570 TI - Effects of laser in situ keratomileusis on the corneal endothelium. AB - The purpose of this study was to assess the effects of laser in situ keratomileusis (LASIK) on the corneal endothelium. In a prospective study, the corneal endothelium of 87 eyes (45 patients) was examined before and 1 month after LASIK. Patients were divided into two groups: people who wear contact lenses (48 eyes) and people who had never worn contact lenses (39 eyes). The corneal endothelium was analyzed for cell density, percentage of hexagonal cells, and coefficient of variation (CV) of cell size. The mean cell density and percentage of hexagonal cells was significantly higher 1 month after LASIK for all 87 eyes. However, the mean CV of cell size was not significantly different. In contact lens wearers, there was a significant increase in mean cell density and percentage of hexagonal cells, but there was no significant change in mean CV of cell size after LASIK. Among patients who had never worn contact lenses, no significant changes were noted in mean cell density, percentage of hexagonal cells, or mean CV of cell size. In this study, LASIK caused no damage to the corneal endothelium. Postoperative improvements in the mean cell density and percentage of hexagonal cells in patients who were contact lens wearers may be related to the discontinuance of contact lens use after LASIK. PMID- 16035571 TI - Abdominopelvic perivascular epithelioid cell tumor with overt malignancy: a case report. AB - Perivascular epithelioid cell tumor (PEComa) is a group of rare tumors composed of epithelioid cells with characteristic perivascular distribution and co expression of the melanogenic marker HMB-45 and muscular markers. There are no documented parameters referring to the biologic behavior of PEComa. We report an abdominopelvic PEComa with overt malignancy in a 16-year-old girl. Histologically, the tumor showed the typical morphophenotypic characteristics of PEComa. Though the cytologic appearance of the tumor cells was relatively bland, the extensive necrosis, presence of lymph node metastases, and surrounding tissue invasion were all indicative of malignancy. Relapse of the tumor with multiple lymphadenopathy shortly after debulking surgery for the primary lesion, and postoperative adjuvant chemotherapy, further denoted its aggressive behavior. PMID- 16035572 TI - Fibroepithelial polyps causing ureteropelvic junction obstruction in a child. AB - Fibroepithelial polyps of the ureter are benign tumors arising from mesodermal tissue in the ureter wall. They are extremely rare lesions that can cause ureteropelvic junction obstruction in children. In this report, we describe an 11 year-old boy with fibroepithelial polyps of the ureter that caused left ureteropelvic junction obstruction. He presented with a 6-month history of left abdominal and flank pain. He also had short stature. Intravenous pyelography showed hydronephrosis without filling defects at the left ureteropelvic junction. Exploration revealed several finger-like polyps obstructing the lumen. This area was resected segmentally and a dismembered pyeloplasty was performed. No complications occurred during the postoperative period. The boy caught up in growth after the operation. Fibroepithelial polyps were confirmed by histology. PMID- 16035573 TI - Conjunctival aspergilloma with multiple mulberry nodules: a case report. AB - A 30-year-old healthy female presented with a 1-year history of chronic mucous discharge, tearing, and irritation in the left eye. Slit-lamp examination revealed severe papillary and follicular reaction surrounding a movable subconjunctival mass on the left upper tarsal conjunctiva. Incision and curettage were performed to establish the diagnosis. Multiple peculiar black mulberry nodules were obtained. The clumps of septate hyphae seen with periodic acid Schiff stain were characteristic of fungus ball (aspergilloma). The patient's symptoms improved significantly after surgery without any antifungal therapy. Although rarely reported, aspergillus is a common fungus in the conjunctiva that may seed into the subconjunctiva. We present this case to remind ophthalmologists of such a rare cause of recalcitrant conjunctival inflammation in immunocompetent patients. PMID- 16035574 TI - Rapid resolution of infantile acute subdural hematoma: a case report. AB - Subdural hematomas in infants are uncommon but usually result from non-accidental trauma or from trauma associated with motor vehicle accidents. This report describes the case of an infant with a traumatic acute subdural hematoma that resolved within 65 hours. A 23-month-old boy fell from a height of approximately 10 m. Brain computed tomography disclosed a left subdural hematoma with midline shift. The associated clots resolved spontaneously within 65 hours of the injury. Although they may mimic more clinically significant subdural hematomas, such collections of clots are likely to be located at least partly within the subarachnoid space. Their recognition may influence decisions regarding both surgical evacuation and the likelihood of non-accidental injury. Clinical and radiographic features distinguishing these "disappearing subdural hematomas" from more typical subdural hematomas are discussed. PMID- 16035575 TI - The participation of minorities in published pediatric research. AB - OBJECTIVES: There is extensive documentation that minority adults are underrepresented in medical research, but there are scant data regarding minority children and their parents. DESIGN: All full-length articles published in three general pediatric journals between July 2002 through June 2003 were collected and reviewed. Articles were excluded if they did not include at least one U.S. researcher, all subjects enrolled at U.S. institutions, parents or children as subjects, some prospective data collection, or between eight and 10,000 subjects. Corresponding authors were surveyed to clarify race/ethnicity data, language barriers and how race/ethnicity data were collected. RESULTS: Two-hundred-twenty eight articles qualified for further analysis. Black children and parents and Asian/Pacific Islander parents were overrepresented, and Hispanic children and parents were underrepresented compared to the Census data. Most researchers collected race/ethnicity data by having subjects self-report. Most studies did not have translation available, although most Hispanic and Asian/Pacific Islander subjects were enrolled in studies in which translation was available. CONCLUSION: Our data show that Hispanic and Asian/Pacific Islander research subjects are more likely to participate in pediatric research when translation is available. If the goal is to ensure access to pediatric research for all ethnic populations, then more research needs to accommodate non-English-speaking participants. PMID- 16035576 TI - Body composition assessment and coronary heart disease risk factors among college students of three ethnic groups. AB - OBJECTIVES: This study identified and compared anthropometric measurements, body composition and coronary heart disease (CHD) risk factors among college students of three ethnic groups. METHODS: Subjects were assessed for cardiovascular risk. Body composition analysis was performed using the Bioelectrical Impedance Analysis (BIA). RESULTS: Black non-Hispanic females (30%) were significantly (p < 0.017) more in the "overweight" category compared to white non-Hispanic females (6.7%). Black non-Hispanic females had significantly (p < 0.044) higher percentages of body fat and lower percentages of body lean, and significantly (p < 0.040) lower percentages of body water than white non-Hispanic females. Significant positive correlations were found between CHD Risk Point Standard (CHDRPS) and percentages of body fat in white non-Hispanic males (p < 0.005), Hispanic males (p < 0.016) and Hispanic females (p < 0.001). Significant inverse correlations were found between CHDRPS and percentages of body water in white non Hispanic males (p < 0.004), Hispanic males (p < 0.013) and Hispanic females (p < 0.001): body lean in white non-Hispanic males (p < 0.005), Hispanic males (p < 0.016) and Hispanic females (p < 0.001); and lean/fat ratio in white non-Hispanic males (p < 0.008), Hispanic males (p < 0.030), black non-Hispanic males (p < 0.020) and Hispanic females (p < 0.008). CONCLUSIONS: The high prevalence of overweight justifies a high priority for weight control in young adults in an effort to prevent cardiovascular diseases (CVDs) later in life. PMID- 16035577 TI - Maternal determinants of pediatric preventive care utilization among blacks and whites. AB - OBJECTIVE: We assessed maternal characteristics that were predictive of preventive care utilization among children 0-5 years and compared black-white differences in preventive care usage. METHOD: We used the 1996-2000 series of public use files from the Medical Expenditure Panel Survey (MEPS). Receipt of preventive care was defined as up-to-date immunization coverage of the child and at least one dental visit during the year. RESULTS: A total of 10,525 children were analyzed consisting of 2,090 blacks (19.9%) and 8,435 whites (80.1%). Black mothers were in general older and less educated. Black households were larger in size and contained a greater number of children. Black mothers earned, on average, much less than their white counterparts even though they tended to be more frequently employed than whites. Despite similar levels of insurance coverage for both racial groups, the overwhelming majority of white mothers were privately insured (73.2%), in contrast to only about half of blacks with private insurance coverage [54.3% (p < 0.0001)]. Overall, the level of pediatric preventive services utilization was a paltry 15.4%. Children of black mothers were significantly less likely to receive preventive care than whites (OR = 0.78; 95% CI = 0.64-0.94). Other determinants of preventive care use were maternal age, insurance coverage, education and family size. Older, educated mothers with insurance coverage and reduced family size were more likely to have their children immunized and have dental visits. CONCLUSION: Maternal characteristics are important markers that indicate the risk for underutilization of pediatric preventive care. A particularly important finding with policy implication is the observation that maternal insurance coverage enhances pediatric preventive care use. Health policy planners may consider parallel insurance coverage of both the child and the mother in order to enhance receipt of preventive health services by the child. PMID- 16035578 TI - Trends in birth across high-parity groups by race/ethnicity and maternal age. AB - BACKGROUND: The changing racial and ethnic diversity of the U.S. population along with delayed childbearing suggest that shifts in the demographic composition of gravidas are likely. It is unclear whether trends in the proportion of births to parous women in the United States have changed over the decades by race and ethnicity, reflecting parallel changes in population demographics. METHODS: Singleton deliveries > or = 20 weeks of gestation in the United States from 1989 through 2000 were analyzed using data from the "Natality data files" assembled by the National Center for Health Statistics (NCHS). We classified maternal age into three categories; younger mothers (aged < 30 years), mature mothers (30-39 years) and older mothers (> or = 40 years) and maternal race/ethnicity into three groups: blacks (non-Hispanic), Hispanics and whites (non-Hispanic). We computed birth rates by period of delivery across the entire population and repeated the analysis stratified by age and maternal race. Chi-squared statistics for linear trend were utilized to assess linear trend across three four-year phases: 1989 1992, 1993-1996 and 1997-2000. In estimating the association between race/ethnicity and parity status, the direct method of standardization was employed to adjust for maternal age. RESULTS: Over the study period, the total number of births to blacks and whites diminished consistently (p for trend < 0.001), whereas among Hispanics a progressive increase in the total number of deliveries was evident (p for trend < 0.001). Black and white women experienced a reduction in total deliveries equivalent to 10% and 9.3%, respectively, while Hispanic women showed a substantial increment in total births (25%). Regardless of race or ethnicity, birth rate was associated with increase in maternal age in a dose-effect fashion among the high (5-9 previous live births), very high (10-14 previous live births) and extremely high (> or = 15 previous live births) parity groups (p for trend < 0.001). After maternal age standardization, black and Hispanic women were more likely to have higher parity as compared to whites. CONCLUSIONS: Our findings demonstrate substantial variation in parity patterns among the main racial and ethnic populations in the United States. These results may help in formulating strategies that will serve as templates for optimizing resource allocation across the different racial/ethnic subpopulations in the United States. PMID- 16035579 TI - Noonan syndrome in a premature infant with hypertrophic cardiomyopathy and death in infancy. AB - We report an extremely premature infant with Noonan syndrome who developed rapidly progressive obstructive hypertrophic cardiomyopathy, which contributed to the death of the infant. The complications of prematurity combined with progressive, severe hypertrophic cardiomyopathy associated with Noonan syndrome lead to a poor prognosis. PMID- 16035580 TI - Screening for cognitive impairment in older adults attending an eye clinic. AB - PURPOSE: We conducted a cross-sectional study examining potentially modifiable factors associated with cognitive impairments (mild or severe) in older whites, African Americans and Hispanics attending an outpatient eye clinic. METHODS: In clinic interviews and physical examinations assessed social, demographic and health information from 100 consecutive Hispanic, African-American and white adults aged > or = 55. Our primary outcome was presence of any cognitive impairment (mild or severe) using the St. Louis University Mental Status Examination (SLUMS) scale. RESULTS: Of the 100 subjects, 65 screened positive for cognitive impairments on the SLUMS cognitive instrument: 46 with mild cognitive impairment and 19 with severe impairment (possible dementia). African-American and Hispanic adults (nonwhites) were significantly more likely to have cognitive impairment compared to white adults (OR 2.80: 95% CI = 1.05-7.44), independent of age, years of education and systolic blood pressure. Subjects with diabetes also had increased odds of cognitive impairments (OR 3.28, 95% CI = 1.21-8.90) even after adjusting for relevant confounders. There was a nonsignificant trend between visual acuity impairment and cognitive impairment (p = 0.059). CONCLUSIONS: Sixty-five percent of adults aged > or = 55 attending the eye clinic screened positive for cognitive impairments, with higher rates among nonwhites and adults living with diabetes. PMID- 16035581 TI - The effects of tea polyphenolic compounds on hair loss among rodents. AB - The objective of this study was to examine the effects of polyphenolic compounds, present in noncommercially available green tea, on hair loss among rodents. In an experimental study, we randomly assigned 60 female Balb/black mice, which had developed spontaneous hair loss on the head, neck and dorsal areas into two equal groups; A (experimental) and B (control). Group A received 50% fraction of polyphenol extract from dehydrated green tea in their drinking water for six months. Group B received regular drinking water. Both groups were fed regular rodent diets (Purina Rodent Chow 5001) and housed individually in polycarbonate cages. The results showed that 33% of the mice in experimental Group A, who received polyphenol extract in their drinking water, had significant hair regrowth during six months of treatment (p = 0.014). No hair growth was observed among mice in the control group, which received regular water. PMID- 16035582 TI - Sickle cell anemia patient with sarcoidosis-associated inguinal lymph node and lung infiltration. AB - Sarcoidosis is a chronic, systemic inflammatory disease of unknown etiology, characterized by noncaseating granulomatous infiltration of any organ. Sickle cell anemia (SCA) is the homozygoid form of sickle cell disease (SCD), which includes a group of genetic disorders characterized by production of an abnormal hemoglobin S (HbS). There are a few case reports with coexistence of sarcoidosis and SCA. We reported a 47-year-old female with SCA and sarcoidosis. PMID- 16035583 TI - Misadventure in traditional medicine practice: an unusual indication for limb amputation. AB - Limb amputation is a major cause of disability in Nigeria, and inadequate health facilities for limb salvage procedures and rehabilitation have increased the burden of amputation surgery in our environment. The common indication for limb amputation in Nigeria is limb gangrene due to trauma. A road traffic accident is an important cause of the trauma, and the role of the traditional bonesetters in the increasing incidence of limb gangrene has been reported. The complications of the traditional bonesetter's practice in Nigeria account for about 50-60% of the limb gangrene necessitating amputation in our hospitals. Misadventures in traditional medicine practice are not new to us. People have lost their lives in the cause of testing the efficacy of traditional medicine. This paper reports an unusual indication for limb amputation in Nigeria following misadventure in a traditional medicine practice in a rural community. PMID- 16035584 TI - Postpartum pyomyoma. AB - Leiomyomata are common benign tumors of the uterus and female pelvis. Myomas have been reported in 25% of Caucasian American women and 50% of African-American women. The true incidence is unknown, but descriptions of 50% have been found at postmortem examinations. Considering the high incidence of uterine myomata in women of reproductive age, they are reported as complications in only 2% of pregnancies. Pyomyoma (suppurative leiomyoma) a rare complication results from infarction and infection of a leiomyoma. Without a strong clinical suspicion and surgical intervention, fatalities may occur. Since 1945, only 15 cases have been described in the literature, mostly in pregnant or postmenopausal women after ascending infection. This report documents a pyomyoma that presented as a postpartum enlargement of a previously known leiomyoma. This case is unique because the patient did not undergo a hysterectomy at the time of exploratory laparotomy. Six months after the procedure, normal cyclic bleeding was noted. PMID- 16035585 TI - Pulmonary alveolar hemorrhage in a pregnancy complicated by systemic lupus erythematosus. AB - We present a case of a pregnant patient with fulminant systemic lupus erythematosus complicated by alveolar hemorrhage, a rare and potentially fatal manifestation of lupus. It typically presents in the context of a pulmonary-renal syndrome. Active lupus nephritis with hypoalbuminemia is a major risk factor for alveolar hemorrhage. Treatment with high-dose corticosteroids is the mainstay of therapy. PMID- 16035586 TI - An unusual odyssey as a physician: an interview with Darrell Pone, MD. Interview by George Dawson. AB - This issue of the Journal of the National Medical Association highlights an interview with Dr. D. Pone. It reveals his journey in becoming a physician. It is both inspiring and quite troubling. It shows the remarkable effort he put into him becoming a practicing physician, from his undergraduate studies to his residency training and, ultimately, his initial practice opportunity. But it also reveals the concerns and travails he experienced while doing so, such as having to change his place of study as an undergraduate and his experience in the OR as a medical student, to name two. PMID- 16035587 TI - Dr. Charles R. Drew: facts, fiction and legend. PMID- 16035588 TI - All humans--"our people," human color scale, affirmative action. PMID- 16035589 TI - Dairy: a re-evaluation. PMID- 16035590 TI - Age- and race-related differences in managing perceived stress. PMID- 16035591 TI - Implementing ICPD: what's happening in countries. PMID- 16035592 TI - Strategy to accelerate progress towards the attainment of international development goals and targets related to reproductive health. AB - Reproductive and sexual ill-health account for 20% of the global burden of ill health for women, and 14% for men. The strategy presented in this document is the World Health Organization's first global strategy on reproductive health. It was adopted by the 57th World Health Assembly (WHA) in May 2004. Five priority aspects of reproductive and sexual health are targeted: improving antenatal, delivery, postpartum and newborn care; providing high-quality services for family planning, including infertility services; eliminating unsafe abortion; combatting sexually transmitted infections, including HIV, reproductive tract infections, cervical cancer and other gynaecological morbidities; and promoting sexual health. The strategy was developed as a result of extensive consultations in all regions with representatives from ministries of health, professional associations, non-governmental organizations, United Nations partners and other key stakeholders. It lays out actions needed for accelerating progress towards the attainment of the Millennium Development Goals (MDGs) and other international goals and targets relating to reproductive health, especially those from the International Conference on Population and Development in 1994 and its five-year follow-up. "The strong endorsement of this strategy by the WHA represents an unequivocal message that countries are committed to do all they can to achieve the goals and targets of the ICPD Programme of Action adopted in 1994." (Dr Paul Van Look, Director, WHO Department of Reproductive Health and Research). PMID- 16035593 TI - Girls can't wait: why girls' education matters and how to make it happen now. PMID- 16035594 TI - Norway at ICPD+10: international assistance for reproductive health does not reflect domestic policies. AB - Norway has a long history of good reproductive health care, with some of the world's best reproductive health indicators. Early reduction of maternal mortality, good services for abortion, contraception and sexually transmitted diseases, a low rate of adolescent pregnancies and a low number people with HIV are examples, achieved through an integrated, publicly provided and funded health care package. Official Norwegian development assistance started in 1952. Emphasis on family planning assistance dates back to 1966, making Norway one of the most consistent donors to family planning and reproductive health programmes. Norway also had a high profile at the International Conference on Population and Development and strongly supported the Programme of Action. Since then, while multilateral support in these areas has stayed high, bilateral support has been downscaled. Overall, international assistance does not reflect the domestic approach to reproductive health services. Norway has given little development support to improvement of maternity services, avoided the issues of abortion and post-coital contraception, and passed up opportunities to support adolescent services. Prevention and treatment of infertility has hardly been an issue. Revitalisation of the reproductive rights discourse in Norway could provide a basis for the protection of reproductive health care domestically, and for policy discussions and decisions in relation to Norway's development assistance. PMID- 16035595 TI - Women's perceptions of reproductive health in three communities around Beirut, Lebanon. AB - The aim of this study was to elicit definitions of the concept of reproductive health among women in three communities around Beirut, Lebanon, as part of the reproductive health component of a larger Urban Health Study. The communities were characterised by poverty, rural-urban mobility and heterogeneous refugee and migrant populations. A random sample of 1,869 women of reproductive age completed a questionnaire, of whom a sub-sample of 201 women were randomly selected. The women's understanding of good reproductive health included three major themes, which were expressed differently in the three communities. Their understanding included good physical and mental health, and underscored the need for activities promoting health. Their ability to reproduce and raise children, practise family planning and birth spacing, and go through pregnancy and motherhood safely were central to their reproductive duties and their social status. Finally, they saw reproductive health within the context of economic status, good marital relations and strength to cope with their lives. These findings point to the need to situate interventions in the life course of women, their health and that of their husbands and families; the importance of reproduction not only from a health services point of view, but also as regards women's roles and responsibilities within marriage and their families; and taking account of the harsh socio economic conditions in their communities. PMID- 16035596 TI - Women in Arab countries: challenging the patriarchal system? AB - Progress in the empowerment of Arab women was found to be low in a 2002 report. Yet Arab women's status is not reflected in continuing high fertility, which in 2000 had dropped sharply in one generation to 3.4. This paper discusses why fertility decline could nevertheless have taken place in the Arab countries. Islam has not stood in the way of fertility decline, as Iran and Algeria show. From the mid- 1970s to 1980s, subsidised consumption through oil wealth redistribution reduced the cost of children, and social conservatism kept married women out of the labour force, both of which promoted higher fertility. The early stages of fertility decline were mainly due to longer length of education of girls, rising female age at first marriage, e.g. 28 in urban Morocco and 29 in Libya, and entry into the labour force of young, single women. There is also a growing population sub-group of never-married young women. Collapsing oil prices and structural adjustment reduced household resources and became an effective fertility regulation factor. Girls born since the 1950s have not only been educated longer than their mothers, but also their fathers, which increases their authority. These factors, and women's activism and civil and political lobbying for the reform of personal status now underway in a number of Arab countries, could all challenge the patriarchal system. PMID- 16035597 TI - The sexual and reproductive health of young people in the Arab countries and Iran. AB - This article reviews the sexual and reproductive health situation of young people aged 10-24 in the Arab states and Iran, based on published and unpublished literature and interviews with 51 key informants working mostly in NGOs and international agencies in the region. There are few national government programmes addressing young people's sexual and reproductive health, with the exceptions of Tunisia and Iran, and a lack of population-based data to guide such programmes. Although the strong emphasis on the integrity and strength of the family unit has a protective effect, young people lack access to information. Education curricula that include these topics are rare and where they do exist, relevant sections are frequently skipped over by teachers, who are unprepared. Health service providers neither recognise the needs of this age group nor make young people welcome, particularly those who are unmarried. Increased education and employment mean the age at marriage is rising, but unprotected forms of marriage are also reported. Taboos surrounding discussion of sexuality remain a key constraint, and data on unwanted pregnancy and abortion, violence against women, and STIs/HIV/AIDS are limited. Building on NGO models and existing efforts, there is a need for the development of national programmes to support the well-being of young people in this region. PMID- 16035598 TI - A strategic assessment of the reproductive health and responsible parenthood programme of Buenos Aires, Argentina. AB - Since 1991, Argentina has had provincial reproductive health laws, a far-reaching national programme and strong public consensus in support of reproductive health policies. Nevertheless, the challenges of strengthening public services, increasing the number of programme sites and resisting conservative attacks remain. This article describes an assessment of the reproductive health programme of the city of Buenos Aires, passed in 2000, whose objectives are to prevent unwanted pregnancies and sexually transmitted diseases/HIV and to train health personnel. The programme operates in every public hospital and primary health care centre in the city. The assessment was conducted jointly by the Ombudsperson's Office of Buenos Aires and the Centre for the Study of State and Society (CEDES). Hormonal contraceptives, IUDs and male condoms were mostly available, but emergency contraception, female condoms and other barrier methods were not Some health professionals and service users were knowledgeable about the new laws and the reproductive rights recognised under the law. Over 90% were satisfied with quality of care in service delivery but many professionals described excessive workloads, deficient infrastructure, and shortages of supplies and staff. Wanting help to obtain a tubal ligation was the most frequent reason for the claims lodged with the Ombudsperson's Office, followed by HIV, quality of care, and abortion. Information and training for both health care providers and women's and human rights NGOs was carried out. PMID- 16035599 TI - The population and reproductive health programme in Brazil 1990-2002: lessons learned. AB - The story of Brazil's evolution in sexual and reproductive health and rights during the 1990s documents not only a decade of change in the population field, but the powerful role of social movements in a democracy. Between October and December 2002, 23 people were interviewed about where they believe progress has been made in Brazil and where there are still needs in relation to population trends, sexual and reproductive rights, and health policies. This paper contains excerpts from the full report and covers the economic and political background of Brazil; the role of non-governmental and women's organisations in influencing the national agenda; the intersection of national and international agendas on population and development, HIV/AIDS, human rights, racism and other issues; changes in sexual and reproductive health policies; HIV/AIDS policy progress, perhaps most importantly mandatory free treatment for people with HIV/AIDS; recent progress in women's health, especially in relation to antenatal and obstetric services, and services addressing violence against women. Finally it describes the role of policy accountability mechanisms that aim to ensure that the many excellent policies that have been passed since 1990 are implemented in a decentralised health system of national, state and local management and services. PMID- 16035600 TI - Sexual and reproductive health and rights in the United Kingdom at ICPD+10. AB - At the mid-point of the Programme of Action of the 1994 International Conference on Population and Development, we reviewed the situation in the UK in key areas. In recent years, greater attention has been paid to sexual health at a national policy level, including strategies on teenage pregnancy and sexual health, but there is still a long way to go. The law against female genital mutilation has been strengthened, twice as many men attend family planning clinics as ten years ago, access to abortion services funded by the National Health Service has improved substantially, up to 80% of contraceptive advice and care is accessed through general practice, a national screening programme for chlamydia is being rolled out, the age of consent has been equalised for men and women and for heterosexuals and homosexuals, and new recommended standards for sexual health and HIV services have been developed. However, many family planning services do not involve young men, waiting times for an abortion are too long, the abortion law has never been extended to Northern Ireland, and there are rising rates of STIs and HIV. Rapid improvements in sexuality education, training and improved resources for family planning and abortion providers and the re-introduction of national public information campaigns about all aspects of sexual health are all required. PMID- 16035601 TI - Global progress in abortion advocacy and policy: an assessment of the decade since ICPD. AB - The 1994 International Conference on Population and Development (ICPD) Programme of Action represented a positive step toward legitimising abortion as a component of basic reproductive health services. This paper reviews how the ICPD principles and recommendations have been applied in the past decade toward increasing women's access to affordable, safe and legal abortion services. It examines advocacy efforts to increase understanding of abortion among policymakers and the public, policy and action at the global level, progress made in national-level policies and services, and obstacles encountered. Research and advocacy are helping to break the silence globally about unsafe abortion, and there is an emerging global movement supporting women's right to safe abortion. A great deal has been accomplished in the ten short years since ICPD, in spite of serious setbacks in some countries and continuing obstacles. A synthesis of public health and rights-based approaches, and strategic partnerships with other social justice movements are called for, as a foundation for effective legal reform efforts and to ensure that women have access to safe abortion services. PMID- 16035602 TI - Orphans and vulnerable children. PMID- 16035603 TI - We are cracking open some taboos--what about the remaining ones? PMID- 16035604 TI - ICPD goals: essential to the millennium development goals. PMID- 16035605 TI - Thinking beyond ICPD+10: where should our movement be going? AB - In this roundtable discussion, three long-time international activists in the field of reproductive and sexual rights discuss the challenges facing the feminist women's health movement ten years after the 1994 International Conference on Population and Development in Cairo, and the 1995 Fourth World Conference on Women in Beijing. Their conversation reflects on the nature of the global context in 1994-95 and how that context has changed a decode later, especially in light of growing conservatism and the growing HIV/AIDS epidemic. They consider the successes and limitations of the Cairo agenda and the strategic importance of taking a long-term view. In particular, they emphasise the need for strengthening health systems worldwide, advocating for better and more woman friendly use of existing resources and building broader coalitions. Much of the conversation explores points of commonality and difference between feminist groups on the one hand, and medical and family planning organisations on the other, as well as ways of utilising such venues as the World Social Forum. Points of greatest promise internationally include the growing youth movement for sexual and reproductive health and rights and the potential for opening up larger alliances around sexual and bodily rights with HIV/AIDS activists, sex workers, people living with HIV and AIDS and human rights organisations. PMID- 16035606 TI - Maternal health and HIV. AB - The HIV/AIDS epidemic is one of the major factors affecting women's health, with 20 million women living with HIV and more than two million pregnancies in HIV positive women each year. Most HIV infections in women are in resource constrained settings where the risk of maternal morbidity and mortality is also unacceptably high, and where most of the 529,000 deaths from complications of pregnancy, childbirth and abortion occur annually. There is increasing evidence that HIV/AIDS-related maternal deaths are escalating considerably, and AIDS has overtaken direct obstetric causes as the leading cause of maternal mortality in some areas of high HIV prevalence. As the availability of antiretroviral treatment becomes more widespread, pregnant women who qualify for antiretroviral treatment should be considered as a priority group for access to treatment. Successful strategies to reduce mother-to-child transmission of HIV are in place in developed countries but much less available in the rest of the world. A more comprehensive approach is needed. The current focus on preventing new infections in children must be broadened to include appropriate care for pregnant women and the prevention of new infections in women and men. PMID- 16035607 TI - Focus on women: linking HIV care and treatment with reproductive health services in the MTCT-Plus Initiative. AB - Despite important advances in expanding access to antiretroviral therapy in the countries most heavily affected by HIV/AIDS, there has been little consideration of the connections between HIV prevention, care and treatment programmes and reproductive health services. In this paper, we explore the integration of reproductive health services into HIV care and treatment programmes. We review the design and progress of the MTCT-Plus Initiative, which provides HIV care and treatment services to HIV positive women as well as their HIV positive children and partners. By emphasising the long-term follow-up of families and the provision of comprehensive care across the spectrum of HIV disease, MTCT-Plus highlights the potential synergies in linking reproductive health services to HIV care and treatment programmes. While HIV care and treatment programmes in resource-limited settings may not be able to integrate all reproductive health services into a single service delivery model, there is a clear need to include basic reproductive health services, such as access to appropriate contraception and counselling and management of unplanned pregnancies. The integration of these services would be facilitated by greater insight into the reproductive choices of HIV positive women and men, and into how health care providers influence access to reproductive health services of people with HIV and AIDS. PMID- 16035608 TI - Syrian women's perceptions and experiences of ultrasound screening in pregnancy: implications for antenatal policy. AB - Ultrasound scanning is firmly embedded in antenatal maternity care around the world. This paper reports on a qualitative study carried out in 2003 of 30 Syrian women's perceptions and experiences of routine ultrasound in pregnancy. It was part of a larger study of the experiences of pregnancy and childbirth of 500 women from Damascus and its suburbs who had recently given birth to healthy newborns. The women had had multiple scans during pregnancy and accepted its use uncritically nearly all the time. The scans gave them reassurance that the baby was healthy, the pregnancy was progressing well and allowed them to learn the sex of the baby. The women also reacted positively to the antenatal educational messages that were conveyed using scans. However, we found the excessive use of this technology worrying. We believe private doctors, who attend 80% of pregnant women, use ultrasound primarily to attract women to their clinics and increase their income. We recommend that maternity care in Syria should be better regulated; that women and their doctors should be made aware of the essential components of antenatal care; that notional guidelines for antenatol care should be developed and that Syrian women should be empowered to ask questions about pregnancy and childbirth and the care they receive. PMID- 16035609 TI - Resumption of sexual relations following childbirth: norms, practices and reproductive health issues in Abidjan, Cote d'Ivoire. AB - Before modern contraceptive methods were available in developing countries, post partum sexual abstinence formed the backbone of birth spacing. With the changes occurring in African societies, how has post-partum sexual abstinence been affected? We conducted an exploratory study in 2000-2001 in Abidjan, Cote d'Ivoire with 23 women and 19 men who were parents of small children. Breastfeeding remains widespread and prolonged. Resumption of sexual relations after delivery was a mean of 11 months. Post-partum sexual abstinence was only distantly related to the traditional lactation taboo. Women expressed fears that their partner would seek elsewhere if they delayed sexual relations too long, and the risk of early pregnancy. Abstinence remained the main way to space births, given low contraceptive use. Mothers generally decided when to wean a child. Men usually made the first move to resume sexual relations, though most women negotiated timing and some insisted on condom use. Provision of condoms post partum can play a contraceptive role for married couples and protect against STIs/HIV in extra-marital relationships, which are frequent post-partum. The duration of post-partum abstinence is in fact unclear because irregular sex may happen early and become regular only later. Women need post-partum information and services that address these issues. PMID- 16035610 TI - Why withdrawal? Why not withdrawal? Men's perspectives. AB - Withdrawal is an ancient and prevalent form of male contraception which has largely been ignored by family planning programme managers all over the world. The objective of this study was to understand men's perspectives on withdrawal use, both users and non-users. In-depth interviews with 62 male factory workers in western Turkey are reported, on use of withdrawal, attitudes to family planning, information on contraception, marital and sexual experience, and gender values and attitudes. Advantages reported both by users and non-users of withdrawal were being free from side effects, ease of access and having no cost. While current users said withdrawal was easy to use and practical, non-users complained about the difficulties of using withdrawal, that it was coitus dependent, caused anxiety and decreased pleasure during sexual intercourse. Current users emphasised taking responsibility as husbands for family planning and protecting their wives from possible adverse effects of contraceptives. Withdrawal should be seen as a valuable choice in a world where increasing method choice, male participation and responsibility taking in reproductive health are all desirable. With its cited advantages withdrawal is likely to fill an important niche among current contraceptive practices and deserves more attention and support. PMID- 16035611 TI - AIDS 2004, Bangkok: a human rights and development issue. PMID- 16035614 TI - Evaluation of head and neck squamous cell carcinoma invasiveness by the electrical resistance breakdown assay. AB - Invasion of tumor cells into the surrounding tissue is a hallmark of cancer. Squamous cell carcinomas of the head and neck region (HNSCC) are characterized by their early primarily lymphatic metastatic spread. The aim of the present study was to evaluate the use of the electrical resistance breakdown assay for determining HNSCC tumor cell invasiveness. The assay utilizes the high transepithelial electrical resistance (TEER) of an epithelial MDCK-C7 monolayer as a sensitive indicator of monolayer integrity and permeability. MDCK-C7 cells were grown to confluence in microfilter membrane cups. 3 x 10(6) cancer cells of cell lines UM-SCC-3, UM-SCC-27, UMB-SCC-745, UMB-SCC-864, UMB-SCC-969 and UT-SCC 26A derived from HNSCC tumors, were seeded on top of this epithelial test barrier. A7-melanoma cells served as a positive control whereas MDCK-C7 cells were used as a negative control and were applied in the same number as the tested tumor cells. TEER was measured over the following days and compared to control values. A significant reduction in TEER was observed in the UMB-SCC-745, UMB-SCC 969 and UT-SCC-26A cell lines within the first 72 h, whereas no significant reduction in TEER was seen in the UM-SCC-3, UM-SCC-27 and UMB-SCC-864 cell lines. HNSCC cell lines in general are found to be less invasive in the resistance breakdown assay compared to other tumor cells such as A7-melanoma cells, however, the electrical resistance breakdown assay appears capable of demonstrating differences in invasiveness between different HNSCC cell lines and therefore potentially could serve as a versatile tool in distinguishing high and low invasive tumors with a potential application as a diagnostic and prognostic marker in clinical investigations. PMID- 16035613 TI - Ovarian carcinoma spheroids disaggregate on type I collagen and invade live human mesothelial cell monolayers. AB - Ovarian carcinoma patients frequently develop malignant ascites containing single and aggregated tumor cells, or spheroids. Spheroids have been shown to be resistant to many therapies, but their contribution to ovarian cancer dissemination remains undetermined. We have previously shown that ascites spheroids adhere to extracellular matrix (ECM) proteins and live human mesothelial cells via beta1 integrin subunits. Here, we assessed the ability of spheroids that were generated from the human ovarian carcinoma cell line NIH:OVCAR5 to disseminate and invade in vitro. Spheroids were seeded on ECM proteins for 24 h. While laminin and type IV collagen stimulated some cell migration, spheroids completely disaggregated on type I collagen substrates. A monoclonal antibody against the beta1 integrin subunit significantly inhibited disaggregation on all proteins tested. To test their invasive ability, spheroids were added to monolayers of live human LP9 mesothelial cells. Within 24 h, the spheroids adhered and disaggregated on top of the monolayers, and within a week had established foci of invasion encompassing a 200-fold larger surface area. Addition of a monoclonal antibody against the beta1 integrin subunit drastically reduced spheroid invasion into the mesothelial cell monolayers. GM 6001, a broad scale matrix metalloproteinase inhibitor, also significantly blocked spheroid invasion into the mesothelial cell monolayers. Epsilon-amino-N-caproic acid, a serine protease inhibitor, partially inhibited spheroid invasion. Based on their ability to attach to, disaggregate on, and invade into live human mesothelial cell monolayers, spheroids should thus be regarded as potential contributors to the dissemination of ovarian cancer. PMID- 16035612 TI - Roles for neuregulins in human cancer. AB - The human epidermal growth factor (EGF) receptor (HER) family of receptor tyrosine kinases has frequently been implicated in cancer. Apart from overexpression or mutation of these receptors, also the aberrant autocrine or paracrine activation of HERs by EGF-like ligands may be important in cancer progression. Neuregulins constitute a family of EGF-like ligands that bind to HER3 or HER4, preferably forming heterodimers with the orphan receptor HER2. Mesenchymal neuregulin typically serves as a pro-survival and pro-differentiation signal for adjacent epithelia. Disruption of the balance between proliferation and differentiation, because of autocrine production by the epithelial cells, increased sensitivity to paracrine signals or disruption of the spatial organization, may lead to constitutive receptor activation, in the absence of receptor overexpression. Consequently, the analysis of ligand expression and/or activated receptors in tumor samples may broaden the group of patients that can benefit from targeted therapies. PMID- 16035615 TI - Syngeneic lymph-node-targeting model of green fluorescent protein-expressing Lewis lung carcinoma. AB - The Lewis lung tumor has been extensively studied in both syngeneic and allogeneic mouse models. However, its metastatic potential and mechanism are poorly understood. The aim of the present study was to develop a highly metastatic lymph-node targeting, imageable model of the Lewis lung carcinoma in a syngeneic host. We report here a syngeneic model of the Lewis lung carcinoma in which the carcinoma cells are labeled with green fluorescent protein (GFP). The tumor cells were transplanted in the dorsal side of the ear of C57-B16 mice in order to give the tumor cells access to the lymphatic system. This model of the Lewis lung carcinoma extensively metastasized to numerous lymph nodes throughout the body of the animal as well as visceral organs, as visualized by fluorescence microscopy using the bright GFP signal. Twenty-one different metastatic sites, including lymph nodes throughout the body, were identified among the cohort of transplanted animals. The data demonstrate a predilection of the Lewis lung carcinoma for lymphatic pathways for metastasis throughout the animal body. The concomitant macrometastases to the visceral organs observed in this model may be remetastasis from the lymph nodes. This model of the Lewis lung carcinoma should be very useful in defining cellular trafficking and targeting mechanisms of metastasis, in particular those involving lymphatic pathways. PMID- 16035617 TI - Caffeine suppresses metastasis in a transgenic mouse model: a prototype molecule for prophylaxis of metastasis. AB - A significant fraction of cancer patients have occult disseminated tumors at the time of primary diagnosis, which usually progress to become clinically relevant lesions. Since the majority of cancer mortality is associated with metastatic disease, the ability to inhibit the growth of the secondary tumors would significantly reduce cancer-related morbidity and mortality. We have investigated whether caffeine, which has been shown to suppress tumor cell invasiveness and experimental metastasis, can suppress metastasis in a spontaneous transgene induced mammary tumor model. Chronic exposure to caffeine prior to the appearance of palpable mammary tumors significantly reduced both tumor burden and metastatic colonization. However, when caffeine exposure began after the appearance of frank tumors, caffeine suppressed metastasis without changing primary tumor burden. The means by which caffeine suppressed metastatic activity may be associated with inhibition of malignant transformation of mammary epithelial cells, inhibition of conversion of dormant tumor cells to micrometastases, micrometastases to macrometastases, or inhibition of tumor cell adhesion and motility. Gene and protein expression patterns resulting from caffeine treatment showed that metastasis suppression may be associated with up-regulation the mRNA expression of multiple extracellular matrix genes, including Fbln1, Bgn, Sparc, Fbn1, Loxl1, Colla1, Col3a1, Col5a1, ColS5a2, ColSa3, Col6a1, Col6a2, and Col6a3. These data suggested that caffeine or other methyl xanthine derivatives may improve the clinical outcome in patients prior to and following the diagnosis of metastatic disease, and could potentially reduce the morbidity and mortality associated with disseminated tumors. PMID- 16035616 TI - Carcinoembryonic antigen promotes tumor cell survival in liver through an IL-10 dependent pathway. AB - Most circulating tumor cells die within 24 h of entering the hepatic microvasculature because their arrest initiates an ischemia-reperfusion (I/R) injury that is cytotoxic. Human colorectal carcinomas (CRC) produce the glycoprotein Carcinoembryonic Antigen (CEA) that increases experimental liver metastasis in nude mice. Since CEA induces release of IL-6 and IL-10, we hypothesized that CEA inhibits the I/R injury through a Kupffer cell-mediated cytokine-dependent pathway. We assessed cytokine effects in CRC co-cultured with liver and in vivo. Human CRC prelabeled with fluorescent dyes were incubated with a reoxygenated suspension of ischemic nude mouse liver fragments in a bioreactor. CEA, rhIL-6 or rhIL-10 were either administered to the donor mice prior to hepatic ischemia or during co-culture. Liver donors were athymic nude or iNOS, IL 6 or IL-10 knock out mice. Ischemic-reoxygenated liver kills Clone A CRC through production of nitric oxide (NO) and superoxide anion. Treatment of liver donors with CEA prior to hepatic ischemia inhibited this in vitro cytotoxicity through an IL-10 and Kupffer cell dependent pathway that inhibited NF-kappaB activation, NO production and iNOS upregulation. IL-10 but not IL-6 enhanced CRC survival in nude mouse liver in vivo. Thus, CEA enhanced metastasis by inducing IL-10 to inhibit iNOS upregulation in host liver. PMID- 16035618 TI - Inhibition of cyclooxygenase-2 suppresses invasiveness of oral squamous cell carcinoma cell lines via down-regulation of matrix metalloproteinase-2 and CD44. AB - Expression of cyclooxygenase-2 (COX-2) in tumors is known to be associated with enhanced angiogenesis, suppression of host immunity, and tumor invasion. In the present study, human oral squamous cell carcinoma (OSCC) cell lines NA and HSC-4 were used to evaluate the effects of NS-398, a selective inhibitor of COX-2, and COX-2 antisense oligonucleotide (COX-2 AS) on the invasion activity of OSCC cells. Matrigel invasion assay revealed that the invasiveness of NA and HSC-4 was suppressed by treatment with either NS-398 or COX-2 AS. These reagents down regulated the secretion of matrix metalloproteinase-2 (MMP-2) to culture supernatant as well as the expression of MMP-2 mRNA and protein. Membrane-type 1 matrix metalloproteinase (MT1-MMP), an activator of proMMP-2, was also down regulated by treatment with these reagents. Furthermore, expression of CD44 on the surface of these cells was reduced by treatment with either NS-398 or COX-2 AS. In addition, MMP-2 antisense oligonucleotides reduced the expression of CD44 on the surface of both OSCC cell lines. These findings suggest that NS-398 and COX-2 AS suppress the invasiveness of OSCC cells via down-regulation of MMP-2 and CD44. Genetic or pharmacological inhibition of COX-2 may therefore be a beneficial strategy in the treatment of OSCC patients. PMID- 16035619 TI - Association of alphavbeta3 integrin expression with the metastatic potential and migratory and chemotactic ability of human osteosarcoma cells. AB - INTRODUCTION: Expression of adhesion molecules such as alphavbeta3 integrin has been associated with the metastatic potential of tumor cells. The purpose of this study was to determine whether alphavbeta3 expression correlated with the metastatic potential of human osteosarcoma cells. MATERIALS AND METHODS: We developed a series of sublines (LM2-LM7) from human osteosarcoma SAOS parental cells, with progressively increasing potential to form lung metastases in nude mice after intravenous injection. SAOS parental and LM2 cells were poorly metastatic, but LM7 cells resulted in visible metastatic lung nodules by 6-8 weeks. We quantified alphavbeta3 integrin expression using flow cytometry. RESULTS: alphavbeta3 expression correlated with the metastatic potential of the cells, with LM7 cells showing the highest expression. LM7 cell adhesion to vitronectin decreased after treatment with echistatin, a RGD-containing peptide antagonist of alphavbeta3. LM7 cells demonstrated higher chemotactic activity than SAOS cells to a homogenate made from lung tissue. This chemotactic activity was also inhibited by echistatin. These data indicated that alphavbeta3 was critical for the migration of LM7 cells to the lung homogenate. Chemotaxis to a liver homogenate was the same for LM7 and SAOS cells. Migration of LM7 cells through lung endothelial cells was higher than that through liver endothelial cells, and echistatin again inhibited this migration. CONCLUSIONS: alphavbeta3 integrin expression may play a role in the metastatic potential of osteosarcoma cells by enhancing the ability of the cells to migrate specifically to the lung. Alphavbeta3 integrin may therefore be a potential new target for osteosarcoma. PMID- 16035620 TI - A high expression level of insulin-like growth factor I receptor is associated with increased expression of transcription factor Sp1 and regional lymph node metastasis of human gastric cancer. AB - Insulin-like growth factor I receptor (IGF-IR) is critical to cell survival and growth and altered IGF-IR expression is found in many human cancers. However, its expression and potential role in gastric cancer development and progression has not been explored. The IGF-IR expression level was determined via immunohistochemistry in primary tumor and lymph node metastasis of 86 cases of resected gastric cancer. Relationships of IGF-IR expression with transcription factor Spl expression and clinicopathological features were analyzed. The impact of altered Sp1 expression on IGF-IR expression and gastric cancer biology was further determined using small inhibitory RNA for Sp1 mRNA. We found that IGF-IR was overexpressed in 62% of the tumor samples when compared with adjacent tumor free gastric mucosa. Patients with lymph node metastases had strong expression of IGF-IR in both primary and metastatic tumor cells. IGF-IR overexpression in the primary tumor correlated with increased lymph node metastasis. Furthermore, the level of IGF-IR expression directly correlated with that of Spl, an important transcription factor for IGF-IR regulation. Knocking-down of Spl expression by small inhibitory RNA led to decreased IGF-IR expression and attenuated growth and metastasis of gastric cancer cells. Therefore, dysregulated expression of IGF-IR and/or Sp1 may contribute to the growth and metastasis of gastric cancer and potentially can be a target of therapeutic intervention. PMID- 16035621 TI - [Bovine leukemia virus as a model for studying human leukemias]. AB - Bovine leukemia virus (BLV) is a retrovirus responsible for lymphoproliferative disorders in ruminants. The comprehensive study of this model system permitted the identification of genetic determinants required for leukemogenesis, the quantification of the dynamic parameters responsible for cell accumulation and the elaboration of novel therapeutic approaches in human. PMID- 16035622 TI - Towards the molecular understanding of the polar overdominance phenomenon associated with the callipyge phenotype in sheep. AB - Most medically or agronomically important phenotypes are "complex" inherited traits. They are influenced by genes but not transmitted according to Mendel's laws. Recently, Steve Henikoff wrote: "The nature of quantitative-trait variation is one of the last unexplored frontiers in genetics, awaiting the future and definitive identification of complex trait determinants, wether they be genetic or epigenetic" (Nature Genetics, 2003). The callipyge phenotype is a muscular hypertrophy in sheep that is characterized by a complex inheritance pattern referred to as polar overdominance: only heterozygous individuals having received the CLPG mutation from their sire exhibit the phenotype. We have spent the last ten years attempting to dissect the molecular basis of this unique phenomenon. These studies have lead to a working model including: (i) DLKI, a paternally expressed growth promoter, (ii) a maternally expressed DLK1 trans-acting repressor that is likely to be a non-coding RNA, and (iii) the CLPG mutation inactivating a silencer element controlling both the expression of DLK1 and its trans-acting repressor in skeletal muscle. This research is not only of fundamental interest, but may as well lead to a better understanding and hence control of "complex" diseases exhibiting "parent-of-origin" effects. PMID- 16035623 TI - [Problems posed by emerging viral diseases in animal health]. PMID- 16035624 TI - [Role of the pharmaceutical industry in the Belgian economy]. PMID- 16035625 TI - [Characterization of new chemoattractant agents in leukocytes]. AB - G protein-coupled receptors constitute one of the largest gene families in mammals. About a hundred orphan receptors still exist, for which the ligands and functions are unknown. We have recently identified the natural ligands of two orphan receptors expressed in dendritic cells and monocytes/macrophages. Chemerin, product of the gene Tig-2, was characterized as the ligand of the chemR23 receptor. The protein is synthesized as an inactive precursor, prochemerin, which requires the proteolytic removal of the last 6 or 7 amino acids, in order to generate a high affinity ligand of chemR23. Two neutrophil proteases, elastase and cathepsin G, are able to mediate this conversion. Besides, a peptide derived from the intracellular protein heme-binding protein (HBP) has been characterized as the first specific ligand of the FPRL2 receptor. The role of these two new systems in the control of physiological and pathological inflammatory reactions is presently being studied. PMID- 16035626 TI - [Gene expression profiling in human thyroid tumors]. AB - Our work aims at defining by microarray gene expression profiles in different thyroid tumors: hyperfunctioning autonomous adenomas, and sporadic and post Chernobyl papillary cancers. Gene expression analysis in hyperfunctioning autonomous adenomas allowed us to identify genes involved in various physiological processes of the thyroid. Concerning papillary cancers, gene expression profiling in post-Chernobyl and sporadic papillary carcinomas could not identify a specific gene expression signature allowing to distinguish both tumors although such a signature exists for autonomous adenomas and carcinomas. This suggests that both cancers represent the same disease, and that there is unlikely to be a molecular signature for radiation-induced thyroid cancer. PMID- 16035627 TI - [Clinical research in Europe and its impact on cancerology--problems and opportunities]. AB - Breakthroughs in molecular biology open new ways forward in therapeutics and subsequently unique opportunities for Europe. Clinical research in oncology is multidisciplinary and very complex. Two distinct objectives should be put forward: 1. To develop, register and market innovative molecules. 2. To develop standard therapeutic strategies. The harmonization of European legislations on clinical trials and financial support to studies conducted without commercial aim pose a major challenge for Europe. Europe should promote clinical research that fosters synergies between universities, healthcare centres and pharmaceutical industry on one hand and public authorities funding healthcare on the other hand, for the best benefit of all patients. PMID- 16035628 TI - [A dart and a microscope for the King--imaginary memoirs of a current microsurgery specialist addressing the Sun King about diseases of the Great Century]. AB - Based on the facts described in King's Louis XIV medical journal, this lecture makes the amazing inventory of the illnesses and the poor state of the medical art in a period when medical empiricism was prevalent. It is, however, the courage of patients and their resistance to utterly absurd and painful treatments that foremost arouse our admiration. By the way, this historical journey also provides the author with a pretext for valorising current developments--in comparison with treatments of the past--such as in the field of microsurgery where some pathologies that were up till now considered to be inaccessible to any possibility of surgical anatomical or functional restoration, are now controlled. PMID- 16035629 TI - [Mechanism and function of dreams]. AB - Man has been fascinated by his dreams for ages. The discovery of rapid eye movement (REM) sleep revived the interest in dream research. The objective study of dream content allowed the characterization of the main features of human dreams: its perceptual content, its pervasive emotional background, its oddity. The particular pattern of cerebral activity observed during REM by functional neuroimaging seems to match these features. Firstly, the perceptual aspects of dreams would be related to the activation of posterior (occipital and temporal) cortices. Accordingly, patients with occipito-temporal lesions may report a cessation of visual dreams imagery. Secondly, emotional features in dreams would be related to the activation of amygdalar complexes, orbito-frontal cortex and anterior cingulate cortex. Thirdly, the activation of mesio-temporal areas would account for the memory content commonly found in dreams. Fourthly, the relative hypoactivation of the prefrontal cortex would explain the alteration in logical reasoning, working memory, episodic memory and executive functions that manifest themselves in dream reports from REM sleep awakenings. Despite these recent results, the precise neural correlates of dreaming remain elusive. Likewise, the functions of dreams are unknown, although usually related to the functions of sleep itself. PMID- 16035630 TI - Advance directives: a call to action. PMID- 16035631 TI - Assessing nursing competency. PMID- 16035632 TI - Antibiotic resistance: the impact on care of hospitalized patients. AB - Incidence of antibiotic-resistant infections is increasing at an alarming rate. The concern about antibiotic resistance has been propelled by increasing questions over the effectiveness of current therapies in the treatment and prevention of severe infections in hospitalized patients. Nurses influence antibiotic resistance in the hospital through infection control procedures, knowledge of evidence-based antibiotic administration protocols, and surveillance of infection incidence and treatments. PMID- 16035633 TI - Huntington disease: a nursing perspective. AB - Huntington disease is a serious neurogenetic disease that affects the physical, cognitive, and psychiatric health of the patient, and has a significant impact on the social life of the family. Nurses play a vital role as health care providers and advocates for the patient with Huntington disease. PMID- 16035634 TI - Managing medication errors--a qualitative study. AB - Medication errors continue to be a significant issue affecting patient safety in United States hospitals. The researchers conducted a qualitative study to explore the understanding and management of medication errors by practicing nurses. The results indicated a belief that late medication administration does not always constitute an error, the use of nursing judgment helps determine when and if medication should be given, and an increased reliance upon computerized and systematic checks put into place in health care systems. PMID- 16035635 TI - Implementing evidence-based nursing practice. AB - A methodology for establishing and supporting evidence-based nursing practice is examined. Description of a clinical and administrative scenario serves as an example of a systematic appraisal of the relevant literature that had implications for clinical practice. PMID- 16035636 TI - Changes in inhaler devices for asthma and COPD. PMID- 16035637 TI - Let's talk 'cancer prevention'. PMID- 16035638 TI - Terri Schiavo and end-of-life decisions: can law help us out? PMID- 16035639 TI - Cytological findings in iron deficiency anemia. AB - Iron deficiency anemia is a common condition in India. We made an attempt to study the cytological changes in the buccal mucosal cells of iron deficiency anemia, patients. Buccal mucosal cells were scraped from 40 iron deficiency anemia patients and were subjected to morphometric analysis. Results of the study showed there is an increase in the cellular diameter, nuclear diameter, and nuclear:cytoplasmic ratio of iron deficiency anemia patients when compared to the normal values. PMID- 16035640 TI - Cytological study of copper in oral submucous fibrosis. AB - BACKGROUND: Habitual arecanut chewing is associated with Oral Submucous Fibrosis (OSF). High copper content in arecanut plays a vital role in pathogenesis of OSF. This study evaluates the copper-staining pattern of buccal epithelial cells in oral cytological smears of non-chewers, chewers, and OSF. MATERIALS & METHODS: Alcohol fixed buccal epithelial smears of 10 histopathologically confirmed cases of OSF, 10 chewers without lesion, and 5 nonchewers were stained with modified Rhodamine technique and studied under the light microscope. Buccal epithelial smears of non-chewers dipped in copper sulphate solution were used as known positive for copper. RESULT: Copper appeared as shades of palered within the cytoplasm of chewers and did not show any stain in non-chewers. Intense red stain was seen in OSF smears as dark granules within the cytoplasm. CONCLUSION: Intense staining of copper in OSF buccal smears, than in the chewers supports the role of copper in the pathogenesis of OSF. PMID- 16035641 TI - Potential for osseous regeneration of platelet-rich plasma--a comparative study in mandibular third molar sockets. AB - Platelet-rich plasma (PRP), the concentrate of platelets in plasma contains various growth factors that enhance osseous regeneration. This study utilized homologous platelet-rich plasma in the treatment of third molar extraction sockets in a total of fourteen human subjects, who were randomly assigned into one control and one test group, each comprising of seven members. Clinical parameters like pain on palpation, number of analgesic tablets consumed, swelling, degree of mouth opening, and condition of mucosa overlying the surgical site were assessed at intervals of 1,3,5,7,9,12, and 16 weeks post operatively. The margins between the socket and surrounding bone, radiopacity of bone filling the socket, and presence oftrabecular bone formation were evaluated simultaneously. Soft tissue healing differed significantly between the two groups, with the test group exhibiting better results. Radiographic evidence of bone formation was visible as early as I week in test subjects. It was concluded that PRP contributed to better healing of soft tissues and bone and is a viable means of growth factor delivery. PMID- 16035642 TI - Oral pH changes due to salivary gland irradiation during radiation therapy of oral and head and neck cancer. PMID- 16035643 TI - Familial occurrence of oral submucous fibrosis: report of eight families from northern Kerala, south India. AB - BACKGROUND: Oral submucous fibrosis (OSF) is an archetype of pathological fibrosis due to possibly its presence for centuries amongst people of Indian ethnic extraction. The WHO criteria for a precancerous condition accords well with this disease. The fact that the disease has been reported in at least few subjects who do not practice the habit of areca nut chewing and the inability to prove a dose-effect relationship in almost all cases, the question arose whether there is a predisposition for the disease. METHODS: A house- to-house survey was carried out in northern Kerala, South India, for the detection of early and advanced OSF from June 2003 through May 2004. An analysis of the family pedigrees of all patients with a positive history of fibrosis has been carried out. RESULTS: Preliminary analysis of the family survey revealed that OSF tends to aggregate in families. The pedigrees of eight (8) such families presented here and the concept of OSF having a genetic predisposition to its occurrence revisited. CONCLUSION: The view that OSF having a family predilection, at least in a proportion of cases, and its tendency for familial linkage seems to be apparent by this study. This rather denovo observation of a trend relationship between OSF risk and the number of putative high-risk genotypes should be separately evaluated for the collagen metabolizing and cross-linking pathways. Establishing familial linkage of the disease seems to be an initial step in deciphering further the molecular pathways involved in its causation and biologic progression. PMID- 16035644 TI - Unusual presentation of nasopharyngeal (juvenile) angiofibroma in a 45 year old female. AB - Juvenile nasopharyngeal angiofibroma is a rare tumour, comprising 0.05% of the head and neck tumours, histologically benign, locally invasive, and has a specific predilection for nasopharynx and adolescent males. This article presents an unusual case of nasopharyngeal angiofibroma in a 45-year-old female patient, manifesting as a destructive maxillary lesion and discusses the two most important factors regarding this tumour, the etio-pathogenesis and spread. PMID- 16035645 TI - Oral intra vascular papillary endothelial hyperplasia in the floor of the mouth. AB - Intravascular papillary endothelial hyperplasia (IPEH) is an unusual vascular lesion of proliferating endothelial cells. It is more frequently seen in the extremities, particularly in the fingers. Oral IPEH has been reported with the common sites being lip, tongue, and buccal mucosa. In this article, we present a case of oral IPEH of the floor of the mouth, an unusual location, presenting in a 9-month-old male. The histogenesis, histologic features, and ultrastructural features are also reviewed. A misdiagnosis of angiosarcoma can be made in a case of IPEH due to similar histopathologic features. It is imperative to rule out this error by an elaborate histopathologic evaluation ofthese lesions. PMID- 16035646 TI - Sturge Weber syndrome. AB - A variant of hemangioma and an uncommon congenital condition, the Sturge Weber syndrome (SWS), also called encephalo trigeminal angiomatosis, is a neurocutaneous disorder with angiomas involving the skin of the face (cutaneous angioma) and pia arachnoid (leptomeningeal angioma). It occurs typically in the ophthalmic and maxillary distributions of the trigeminal nerve (1,2). Here we present a case of a 14 year old girl. PMID- 16035647 TI - Tyrosinase induction and inactivation in normal cultured human melanocytes by endothelin-1. AB - Since endothelin was found to be expressed in epithelial cells as well as in vascular endothelial cells, the functional regulation of melanocytes with endothelin has been actively investigated. In particular, it has been suggested that endothelin may influence pigmentation and depigmentation, which are mediated by melanocytes. In the present study, we investigated the regulation of melanocyte function and tyrosinase expression by endothelin from the point of view of tyrosinase protein expression and enzyme activity. The influence of endothelins on melanocyte function was assessed. Melanocytes showed a dose dependent increase in cell proliferation with the addition of endothelin-1. When the confluence of melanocytes was cultured with endothelin-1 for 72 h, tyrosinase activity in melanocytes was significantly and dose-dependently decreased. In contrast, there was no significant change with endothelin-3. However, tyrosinase protein expression of melanocytes was significantly and dose-dependently increased by endothelin-1, but endothelin-3 had no effect. Both the suppression of enzyme activity and the enhanced protein expression were regulated by the ETA receptor antagonist, BQ123. In view of these observations, we conclude that endothelin-1-induced tyrosinase is mediated by ETA receptors. However, the reason for the decrease in the specific activity of tyrosinase remains unknown, and our results suggest that another mechanism underlying the activation of tyrosinase is present in addition to the inductive action of endothelin-1 on tyrosinase. PMID- 16035648 TI - Distribution of aquaporin-9 in the rat: an immunohistochemical study. AB - The aim of this study was to identify the distribution of aquaporin-9 by immunohistochemistry in rat tissues using specific antipeptide antiserum which we recently produced. Anti-aquaporin-9 antibody was raised in New Zealand white rabbits immunized with a conjugate of synthetic aquaporin-9 peptide with bovine serum albumin. Immunohistochemical analysis was performed by avidin-biotin complex method. Aquaporin-immunoreactivity was visualized in the anterior pituitary, central nervous system, retina, thyroid gland, gastrointestinal tract, liver lung, pancreas and testis. When using antiserum preincubated with synthetic peptides or rat hypothalamus homogenate, which contains aquaporin-9 peptide, no significant stain of the hypothalamus was detected. These findings suggest that aquaporin-9 is widely distributed and that the method used is valuable in studying the distribution of aquaporin-9 in rats. PMID- 16035649 TI - Otsuka Long-Evans Tokushima fatty (OLETF) rat is not a suitable animal model for the study of angiopathic diabetic retinopathy. AB - We have previously shown that the peak latency of oscillatory potential (OP), the earliest electroretinographic manifestation of diabetic retina, was prolonged in Otsuka Long-Evans Tokushima Fatty (OLETF) rat, a model of non-insulin-dependent diabetes. These observations suggest that retinal neuronal dysfunction revealed by the OP abnormality in the electroretinogram takes place prior to the angiopathic diabetic changes in this animal model. However whether acellular capillaries and pericyte ghosts, one of the histopathological hallmarks of early diabetic retinopathy in humans, could occur in OLETF rat remains to be elucidated. In the present study, we first prepared the retinal trypsin digests of OLETF and control Long-Evans Tokushima Otsuka (LETO) rats at 45 weeks old and then compared the number of acellular capillaries and pericyte ghosts in the retinas of OLETF rats with that in LETO rats. Blood glucose levels were higher in the OLETF rats than those in LETO rats. Retinal capillaries of OLETF rats were found to remain morphologically normal and pericyte ghosts were barely detectable. There was no difference in the number of acellular capillaries in the retinas between OLETF and LETO rats. The present study indicates that acellular capillaries and pericyte ghosts, the characteristic morphological changes in early diabetic retinopathy, are not accelerated in OLETF rats. Our data suggest that OLETF rat is not a suitable animal model for the study of angiopathic diabetic retinopathy. PMID- 16035650 TI - Atheroprotective properties of nifedipine. AB - Impaired endothelial cell growth and function have been suggested to be an initial event leading to the development of atherosclerosis. Nifedipine is one of the most widely used dihydropyridine-based calcium antagonists (DHPs) for the treatment of patients with angina and hypertension. Recently, nifedipine was shown to improve endothelial function in many cardiovascular diseases, thus slowing the development and progression of atherosclerosis. In this review, we discuss the molecular mechanisms for the atheroprotective effects of nifedipine, with special emphasis on its anti-oxidative properties. PMID- 16035651 TI - The effects of rofecoxib on 24-h ambulatory blood pressure and heart rate monitoring in patients with hypertension and osteoarthritis. AB - The aim of the study was to investigate the effects of the cylooxygenase (COX)-2 specific inhibitor rofecoxib, on blood pressure (BP) and heart rate (HR) in patients with well-controlled hypertension and osteoarthritis via 24-h ambulatory monitoring. Thirty patients with well controlled hypertension were included. Fifteen patients had osteoarthritis and were recommended by their rheumatologists to take rofecoxib 12.5 mg/day (rofecoxib group). The control group consisted of 15 patients who had hypertension but no clinical osteoarthritis and did not receive any anti-inflammatory drugs. Twenty-four-hour ambulatory monitoring of BP and HR were performed on the day before initiation of rofecoxib therapy and on days 3 and 14 of COX-2 therapy. The control group underwent 24-h monitoring three times at similar intervals. Antihypertensive medications were continued. On day 3 of rofecoxib therapy, mean HR for both daytime and nighttime were lower than those at baseline. On day 14, the changes in mean HR did not differ from baseline values. Similarly, diastolic BP (daytime and nighttime) on day 3 appeared to be lower than at baseline. However this difference was not observed on day 14, and mean daytime and nighttime diastolic BP returned to baseline values. There was no statistically significant difference in the mean arterial pressure or systolic BP recordings on days 3 or 14 than at baseline. Rofecoxib 12.5 mg/day did not significantly increase BP during 24-h ambulatory BP monitoring in patients with well-controlled hypertension and osteoarthritis. PMID- 16035652 TI - Feasibility study of nitrogen removal with the mecellulose wasted liquor as an external carbon source in the two-stage denitrification process. AB - The utilization of mecellulose wasted liquor (MWL) as an external carbon source was investigated to find an alternative for methanol in the two-stage denitrification pilot process. The pilot plant was supplied with the raw water from the J-Municipal Sewage Treatment Plant (J-MSTP) in Korea. The raw water of J Municipal Sewage Treatment Plant contains low and high concentration of biodegradable organics and nitrogen source, respectively, due to the inflow of industrial wastewater and landfill leachate. Methanol was fed to provide external carbon source for high concentration of nitrogen source removal by denitrification in this J-Municipal Sewage Treatment Plant, and thus this study was performed to test effects to the effluent quality and efficiencies of nitrogen source removal with an alternative carbon source for the cost reduction. The 6.5mg 1(-1) and 5.7mg l(-1) of total nitrogen (TN) concentration in the effluent were achieved with mecellulose and methanol, whereas mecellulose and methanol were fed to give the same ratio of gCODgNO,-N(-1), respectively. The 60% of COD in mecellulose wasted liquor was used as a carbon source for denitrification and the stable denitrification rate was earned when one half of the required total carbon source for denitrification was fed to pre-anoxic tank in the pilot plant. The required gCODgNO,-N(-1) ratio with mecellulose wasted liquor was 1.4 times higher than with methanol. Mecellulose wasted liquor is feasible to be used as external carbon source for organic loading, nitrogen and phosphorus removal. If mecellulose wasted liquor is considered as an alternative external carbon source to substitute methanol 26-28m3 mecellulose wasted liquor per 1 m3 methanol will be required. However, to meet with the effluent standard (10 mg BOD l(-1)) for J-Municipal Sewage Treatment Plant, the feed concentration of mecellulose wasted liquor should be recommended to be lower than 200 mgl(-1). PMID- 16035653 TI - Estimating biomass of heterotrophic and autotrophic bacteria by our batch tests. AB - A new method was developed, carrying out the batch tests of oxygen utilization rate with and without the addition of Allythiourea solution (inhibitor) at the same time, to separately and simultaneously estimate both the biomass of heterotrophic and autotrophic bacteria in a mixed culture activated sludge of a sequential batch reactor system. The results of the batch tests showed that the YH kinetic parameter was equal to 0.77 g cell COD per g oxidized COD and the maximum specific growth rate and biomass of heterotrophic bacteria were 2.13 d( 1) and 212 mg l(-1) as COD (about 66% of the activated sludge), respectively. On the other hand, the maximum specific growth rate and biomass of autotrophic bacteria were 1.62 d(-1) and 2.51 mg l(-1) as COD (about 0.8% of activated sludge), respectively. The proposed method is easier and cheaper than other developed methods. PMID- 16035654 TI - Metal removal from wastewater by bacterial sorption: kinetics and competition studies. AB - Biosorption has distinct advantages such as high efficiency and cost effectiveness for removal of metal ions from wastewater. However, most studies of bacterial biosorption have focused on the removal of a single metal ion from aqueous solutions, even though natural water and wastewater rarely contain only one kind of heavy metal. This study investigated competitive biosorption of Co2+, Ni2+ and Cr3+ in different combinations at various concentrations under conditions of co-existence in a synthetic wastewater system utilizing the gram negative bacterium Pseudomonas aeruginosa. In a binary system with Co2+ and Ni2+, the presence of the same concentration of dissolved Ni2+ led to a significant decrease in the amount of Co2+ adsorbed onto cells when compared with a single Co2+ system in which Ni2+ was absent. This result is most likely due to the similar competitive affinity of these equivalent cations for sorption sites located on the cells of the bacterium. However, the presence of Cr3+ with either Co2+ or Ni2+ led to a strong reduction in the uptake of each of these bivalent ions by cells, whereas Cr3+ adsorption was unhindered. Cr3+ also significantly inhibited the adsorption of both Co2+ and Ni2+ onto cells in a ternary system involving Co2+-Ni2+-Cr3+, with the order of preferential metal uptake being Cr3+ > Co2+ approximately Ni2+. Removal efficiency of the heavy metals reached 100 % at low concentrations ([Cr3+]=[Co2+]=[Ni2+=20 microM) in a multi-metal system. The results of this study indicate that pre-determination of co-existing heavy metals in wastewater and examination of their competition for the sorption sites of biosorbents are required for the efficient removal of inorganic toxicants utilizing biosorption processes. PMID- 16035655 TI - The effects of post-treatments and temperature on recovering the methane potential of >2 mm solid fraction of digested cow manure. AB - The effects of thermal and chemical treatments, mechanical maceration and freezing and thawing on recovering the remaining methane potential of the >2 mm solid fraction of digested cow manure - which accounted for 30% of the original potential of digested cow manure - were studied in laboratory batch assays at 5 20 degrees C and at 35-55 degrees C to evaluate the treatment effects both under long-term (340 d) storage of solids and during active digestion (30 d), respectively. The effects of different treatments on the methane production of the solids varied with incubation temperatures and time. However, in all cases, methane productions at 15 degrees C and lower were slow and low for both untreated and treated solids even after long-term incubation. At 35 and 55 degrees C more methane was recovered from untreated solids producing up to 61-82 ml g(-1) volatile solids (VS)added in 30 d and 179-215 ml g(-1) VSadded in 340 d. Only chemical treatment with or without thermal treatment enhanced the methane yields while some treatments even decreased the yields. An increase in temperature to 35 degrees C of the assays incubated for 6 months at < or =20 degrees C initiated more significant methane production. In conclusion, the methane potential of the digested solids in a farm-scale biogas system can be recovered by active digestion at 35 or 55 degrees C and can be improved to a smaller extent through chemical treatment of separated solids fraction, while methane recovery at lower temperatures and with some of the treatments studied would not be effective. PMID- 16035656 TI - Olive oil mill wastewater purification by combination of coagulation- flocculation and biological treatments. AB - In order to define an efficient pre-treatment of Olive Oil Mill Wastewater (OOMW) to overcome major obstacles to biological treatment, various organic and mineral coagulants have been tested. In particular, the application of quicklime until a pH around 12 - 12.4 was reached, allowed the reduction of almost 37% of the initial COD, and approximately 88% and 71% of the colour and phenolic content of the waste. Hence, further biological treatments with an adapted aerobic consortium (AC) and a white rot fungus (WRF) strain were improved. The WRF Coriolopsis polyzona was more efficient than AC to reduce colour and polyphenols when the waste was prior diluted or pre-treated; however, it was less effective in COD removal. The combined treatment: lime - AC of OOMW having initial COD of 102 g l(-1) led to the elimination of about 77, 91 and 63%, of the COD, phenols and colour, respectively. Interestingly, the opposite combination AC - lime permitted better COD, phenols and colour reduction to respectively, 21, 11 and 11% of the initial values. This latter condition is technically recommended since only one step separation was needed and no pH correction was necessary before undergoing aerobic treatment. Moreover, the process would produce a sludge potentially rich in organic matter, and consequently, useful as an agricultural amendment or/and as an additive in animal nutrition. PMID- 16035657 TI - Black shale as a sorbent for trichloroethylene and Cr(VI). AB - Black shale was examined as a natural sorbent for organic and inorganic contaminants. Trichloroethylene (TCE) could be removed well from the water by sorption onto the locally available black shale because of the high organic carbon content (5.2%) of the black shale in this study. Hexavalent chromium Cr(VI) was mainly removed by ionic sorption and reduction in batch and column experiments. Amphiphilic humic acid was also sorbed onto the black shale and could facilitate the sorption of TCE at the same time. Humic acid also enhanced the removal of Cr(VI) by reduction and sorption, but the amount of Cr(VI) adsorbed (mg kg(-1)) was smaller than that of TCE. Considering that the black shale in this study was used without any modifications and has a small surface area, black shale can be a cost-effective natural geosorbent and additive to remove organic contaminants and heavy metals. PMID- 16035658 TI - The influence of ferrous/ferric ions on the efficiency of photocatalytic oxidation of pollutants in groundwater. AB - The complex influence of ferrous/ferric ions on the efficiency of aqueous photocatalytic oxidation (PCO) of 2-ethoxyethanol (2-EE), methyl tert-butyl ether (MTBE) and humic substances (HS) was established. A drastic efficiency increase at lower concentration of ferrous/ferric ions was observed to change to a sharp decrease at higher concentrations for 2-EE and MTBE, whereas for HS only an inhibitive effect of Fe2+/3+ on the PCO efficiency was noticed. The authors proposed an explanation for the observed phenomena based on the different sensitivities of pollutants towards radical-oxidation reactions and the competitive adsorption of metallic ions and pollutants on the TiO2 surface. PMID- 16035659 TI - Transformation of chlorinated methanes by zero-valent aluminum coupled with Pd/Al2O3. AB - The transformation of chlorinated methanes by zero-valent aluminum under various pH conditions has been examined in the presence and absence of Pd/Al2O3. In the absence of Pd/Al2O3, carbon tetrachloride was primarily transferred to chloroform (65%) and dichloromethane (15.3%) at pH 2.5 while little of the hydrodehalogenation intermediates (approximately 5%) was found at pH 10.8. This indicates that carbon tetrachloride undergoes hydrodehalogenation at low pH but not at high pH. The low production of hydrodehalogenation intermediates at high pH could be attributed to the base-catalyzed hydrolysis of carbon tetrachloride resulting in the formation of products that can not be detected by the provided analytic methods. In the presence of Pd/Al2O3, degradation rates were increased by a factor of 10-43 compared to the system without Pd/Al2O3, suggesting Pd/Al2O3, serves as a catalyst. Further, complete degradation of carbon tetrachloride by zero-valent aluminum with Pd/Al2O3 led directly to methane (86.4%) without formation and accumulation of dichloromethane at pH 9.0. The direct transformation of carbon tetrachloride to methane can be rationalized by the catalytic hydrodehalogenation occurring at the palladium surface. The study indicates aluminum coupled with Pd/Al2O3 is an environmentally-friendly technology for treatment of chlorinated organic contaminants. PMID- 16035660 TI - Extraction behavior of metallic contaminants and soil constituents from contaminated soils. AB - With an aim of developing an effective remediation technology for soils contaminated by heavy metals and metalloids, the extraction behavior of metallic contaminants as well as those of soil constituents was studied on a laboratory scale. Three contaminated soils collected from a former metal recycling plant were examined. These three soils were found to be contaminated by As, Cu, Pb, Sb, Se and Zn as compared to the non-contaminated soil. The pH-dependent extraction behavior of various elements from the soils was measured in a wide pH range and categorized into three groups. Hydrochloric acid (HCl), H2SO4, H3PO4, HNO3, sodium citrate, sodium tartrate, disodium dihydrogen ethylenediaminetetraacetate and diethylenetriaminepentaacetic acid were evaluated as extractants for removing contaminants from the soils. Extraction behavior of the soil constituents was also studied. The efficiency of the extraction was evaluated by the Japanese content and leaching tests. The stabilization of Pb remaining in the soil after the extraction process was conducted by the addition of iron(III) and calcium chloride. PMID- 16035661 TI - [Experimental validation of calcium phosphate precipitation modeling in a pellet reactor]. AB - Calcium phosphate precipitation is studied in this article. The P-recovery process is carried out in a fluidized sand bed, the so-called pellet reactor which presents major advantages from the hydrodynamical viewpoint. The associated chemistry is yet relatively complex, due to pH gradient along the column and to the residence time of the various precipitates. The experimental observations showed three different phenomena: first, an agglomeration of fines around the sand grains is observed, second, a stagnation of fines in the bed occurs while a significant amount of fines also leaves the bed with the liquid effluent. The purpose of this work is to validate the thermodynamical model developed in our previous works on a semi-industrial sized pilot. Additional experimental runs carried out for various operating conditions showed the robustness of the model. These results open some interesting perspectives for the determination of optimized operating conditions at industrial scale. PMID- 16035662 TI - Experimental evaluation of inhibition effects of saline wastewater on activated sludge. AB - Industrial wastewater often has a high salt content, due to specific factories and to the use of seawater in coastal areas. Once it has reached the activated sludge treatment, saline wastewater may cause biomass inhibition and dramatic loss in purification efficiency. In the present work, the inhibiting effect of saline influent was experimentally evaluated through respirometric tests on samples of the incoming wastewater. Activated sludge was grown in bench-scale continuous flow stirred tank reactor (CFSTR), supplied with synthetic wastewater. Synthetic saline wastewater was prepared using sodium chloride and sodium sulphate at different concentrations. Samples of mixed liquor were drawn from the CFSTR reactor and fed with saline wastewater in shock-load or in continuous mode, then respiration activity and carbon removal yield were evaluated. In batch mode (shock-load), salt/biomass ratios between 0.37 and 30.7 gsalt gvss(-1) gave respiration inhibition between 4% and 84% respectively. The respirometry bioassay required a short response time and the values of respiration inhibition predicted the loss in carbon removal efficiency with a good accuracy. In continuous mode, for salt/biomass ratio of 35.5 gsaltgvss(-1), respiration inhibition of 81% was obtained. This work showed that a relation between inhibition in shock-load and inhibition in continuous mode can be determined, therefore, the procedure herein proposed allows performing offline bioassays on real biological plants, to assess in real time the inhibiting effect of the incoming wastewater before it reaches the activated sludge reactor. PMID- 16035663 TI - Treatment of textile wastewater using a natural flocculant. AB - The physicochemical treatment of wastewater is of substantial interest, especially when conventional treatments by biological processes are not amenable. Among the current chemical processes used for industrial wastewater treatment, coagulation-flocculation has received a large attention for high pollutant removal efficiency. This paper summarizes the results of a textile wastewater treatment process aimed at the destruction of colour by coagulation-flocculation process and using an organic natural flocculant: tannic substances. Jar-test experiments were carried out in order to determine the optimum conditions for the removal of organic matter and color. Treatment with studied flocculent (Polysep3000) proved to be effective in a wide pH range (pH < or = 10), very effective in removing the color (96 %) and a fraction of the COD (40-50 %) from the textile wastewater. Polysep3000 is compared to Al2 (SO4)3 and FeCl3 for sludge production. Polysep3000 seems to be suitable for yielding a high removal of color (> 90%) and corresponding low volume of settled sludge. PMID- 16035664 TI - [Short account of the history of thyroid surgery]. AB - Historical writings have described abnormalities of the thyroid gland for more than 3500 years. The relationship between the thyroid gland and goiter has been unknown until the XX century. Originally, what we know today as goiter was described as bronchocele. It was Wharton who in 1656 named the gland "thyroid", not because of the shape of the gland but because of the proximity to the thyroid cartilage. Rare attempts at thyroidectomy were made early on primarily for prevention of suffocation secondary to goiter with little success and a mortality rate as high as 40%. In 1791, Desault performed the first successful partial thyroidectomy. The most notable surgeon of 20th-century was Emil Theodore Kocher, who is considered the father of thyroid surgery. Christian Albert Theodor Billroth also made significant contributions to thyroid surgery. PMID- 16035665 TI - [Benign thyroid nodule: what is the treatment? Personal experience]. AB - INTRODUCTION: Solitary thyroid nodule is an important pathology with an incidence of 2-5% in the Italian population. The diagnosis was based on clinical exam, laboratory tests and ultrasonographic evaluation. MATERIALS AND METHODS: The study undertook has the purpose to assess the usefulness between lobe isthmusectomy (with corresponding risk of possible relapse and complications for reintervention) and total thyroidectomy (with corresponding post-surgical treatment complications), for benign thyroid disease with solitary nodule, on selected cases for disease and corresponding risk factors, in the retrospective clinical study, using laboratory tests and ultrasonographic evaluation. The study was conducted on 80 patients admitted from 1994 to 2000 with diagnosis of benign thyroid nodule and operated with lobe-isthmusectomy. In seven patients the operation had to be converted in total thyroidectomy. RESULTS: Operative mortality was nil and long-term results at a mean follow-up of three years are encouraging. DISCUSSION: The main advantages of lobe-isthmusectomy for benign solitary thyroid nodule consist in less postoperative complications and less hospital stay. CONCLUSIONS: Therefore our experience has carried us to consider the lobectomy which treatment of choice for all those benign thyroid diseases with solitary nodule. PMID- 16035666 TI - [Surgical treatment of Graves' disease: results in 108 patients]. AB - AIM OF THE STUDY: To evaluate morbidity and functional results of surgical treatment in patients with Graves' disease. METHODS: A retrospective study was performed in 108 patients operated on during 1993-2003. Main indications for surgery were failure of treatment with antithyroid drugs (80.6%), large goiter (46.3%) and/or severe ophthalmopathy (23.1%). Surgical procedures were extensive subtotal thyroidectomy (EST; n = 33; uni- or bilateral remnant of <2 g) or total thyroidectomy (TT; n=75). Functional results were established in 89 patients (27 EST patients and 62 TT patients) after a mean follow-up of 5.9 years. RESULTS: Operative mortality was zero. There were 4 (3.7%) transient unilateral recurrent laryngeal nerve (RLN) palsies and no cases of permanent RLN palsy. Temporary hypocalcemia occurred in 15 patients (13.9%) and permanent hypoparathyroidism resulted in two (1.9%). Four patients (3.7%) developed a postoperative hematoma that required reoperation. There was no significant difference in the rate of complications between EST and TT, although temporary hypocalcemia was more common following TT than EST (17.3% vs. 6.1%) and permanent hypoparathyroidism affected only TT patients. None of the patients developed recurrent hyperthyroidism; all patients are maintained on levothyroxine. CONCLUSIONS: Surgery is an effective therapy for selected cases of Graves' disease. When performed by experienced surgeons, it can be carried out with no mortality and minimal morbidity. EST (with uni- or bilateral remnant of <2 g) and TT are both effective in order to achieve a definitive cure of hyperthyroidism. PMID- 16035667 TI - [Hyperthyroidism in the elderly]. AB - Hyperthyroidism in elderly patients is not to be under-evaluated, since it is characterized in such age range by particular clinical and prognostic features. Based upon literature survey and their clinical experience, the authors discuss in the present paper clinical, diagnostic and therapeutic problems of hyperthyroidism in geriatric patients. MATERIAL AND METHOD: In the period between 1978-2003 out of 1804 patients surgically treated for thyroid disease (non neoplastic in 1470 pts.), 180 subjects presented hyperthyroidism (17%). 36 were in geriatric age-range (mean age 76 yr.; 29 females and 7 males). 26 presented a Multinodular Toxic Goiter (72.2%), whereas 9 patients complained of Plummer Adenoma (25%); only 1 patient showed Basedow disease (2.7%). RESULTS: As far as ASA classification, there were 7 ASA I, 27 ASA II and 2 ASA III. Compression of digestive tract and/or respiratory airway represented a surgical indication in 15 patients (41.6%). 12 (33.4%) were operated due to predominant cardiac symptoms (tachycardia, atrial fibrillation). The remaining 9 patients (25%) were treated for the concomitance of atypical symptoms of hyperthyroidism. We performed 15 total thyroidectomy, 7 "near totally", 8 sub-total, 6 hemithyroidectomy in case of Plummer adenoma. Postoperative mortality was nihil; p.o. morbidity was 5.5% for medical conditions (pneumonia) and surgery-related (1 laryngeal recurrent paralysis and 1 hypoparathyroidism) in 5.5%. Post-operative follow-up, conducted at 6 and 12 months from the operation, showed regression of hyperthyroidism and regression or improvement of all clinical symptoms complained by the patient. CONCLUSION: Surgical treatment seems to be the only immediate and definitive cure for hyperthyroidism. Geriatric age does not seem to be a surgical contraindication. PMID- 16035668 TI - [Prevention of complications in thyroid surgery recurrent laryngeal nerve injury personal experience on 313 cases]. AB - INTRODUCTION: Thyroidectomy poses many challenges for the surgeon who undertakes endocrine surgery and iatrogenic injury of inferior laryngeal nerve (ILN) is one of the most serious (0-20%). We report our personal experience of a series of 313 thyroidectomy with intraoperative identification of ILN. METHODS: 313 patients (253 females, 60 males, whose age was between 17 and 86 years, mean 41 years) had undergone thyroidectomy in our Operative Unit from January 2000 to January 2004. Among them, 259 patients underwent total extracapsular thyroidectomy, 38 subtotal thyroidectomy, 5 isthmo-lobectomy and 11 were completions of thyroidectomy in patients who had previously undergone a first thyroid surgical intervention. RESULTS: We identified 588 ILN (in all cases), in the left or in the right side only in case of isthmo-lobectomy or completion of thyroidectomy. In two cases (0.63%) we noticed on the right side a non recurrent laryngeal nerve. Concerning the postoperative results we noticed only one case (0.38%) of ILN injury with monolateral vocal cord hypomotility and temporary dysphonia, actually on phoniatric therapy at follow-up. CONCLUSION: A strong knowledge of the anatomy and embryology of the thyroid region, a commitment to meticulous attention to detail, the awareness of the extremely varying course of the ILN and the inferior thyroid artery and their relations, and adequate experience are all required to maintain a level of expertise and avoid ILN. Thyroid carcinoma, recurrent goitre, total thyroidectomy operation, duration of the operation are factors which increase the risk of postoperative ILN injury. PMID- 16035669 TI - [Identification of an objective reference parameter for the reconstruction of a large incisional hernia following physiological principles]. AB - Repairing a parietal defect of a large incisional hernia should not be limited to the closure of the breach by means of the modern biocompatible prosthetic sheets, but must also be able to restore a correct intra abdominal pressure, otherwise the derangement from the normal respiratory dynamics and the circulatory stasis in the abdominal veins and in those of the lower limbs remain unaltered. Over correcting the parietal abdominal tension on the contrary can cause a dangerous compartmental syndrome. The attempts of an intraoperative measurement of the correct intra abdominal pressure restoration has been generally hampered from the condition of curarization of the patient during the operation. Using the automatic mechanical ventilator fixed at volume and not at pressure priority, as usual, can offer the possibility to calibrate, following objective parameters, the propriety of the surgical repair still during the final phases of the reparative operation. The simplicity and ingenuity of the here proposed method and the normal availability in every operative theatre of the necessary means for this measurements described, requires attention among the surgeons and a large diffusions of its simple use. PMID- 16035670 TI - The possible role of radiofrequency as complementary treatment of locally advanced gastric cancer. AB - AIM: The prognosis of locally advanced gastric cancer (T3-T4) is bad. The presence of lymph nodes (N3-N4) or haematogenous metastases (liver, lung) gets worse the evolution; principally the hepatic malignancies are cause of scarce survival. The possible use of a palliative treatment as radiofrequency ablation (the good results are note about the treatment of hepatic malignancies by colo rectal cancer) is reported in recent series. Therefore we decide to use radiofrequency ablation for the treatment of hepatic metastases by gastric cancer, difficulty treated surgically. MATERIALS AND METHODS: From January 2001 to December 2002, 25 patients affected by hepatic metastases underwent to radiofrequency thermal ablation, 2 of them were affected by gastric adenocarcinoma. Case 1: A.P., 58 year-old man, one year before underwent to subtotal gastric resection according to Billroth II. After repeated postoperative chemotherapy cycles, he presented metastases at IV hepatic segment. The patient underwent to percutaneous radiofrequency ablation. The control CT scan confirmed metastasis disappearance. After three months, a partial recurrence was treated by the alcoholization. Three months after, we observed marked jaundice for multiple diffused metastases, followed by the exitus. Case 2: B.G., 63 year-old man, with advanced gastric adenocarcinoma (T4) at the pylorus and hepatic metastasis at IV segment. The patient underwent to gastrojejunostomy and to intraoperative radiofrequency ablation. Ultrasonography and CT scan controls were performed before discharge. The patient didn't undergo to successive controls. After 7 months, the patient returned with marked jaundice for diffused hepatic metastases; he refused any treatment, and then he died one month after. CONCLUSIONS: Our preliminary results don't show complications related to the intra and peri-operative radiofrequency, with an important increase of the mean survival. The results, limited by poor experience, may indicate the complementary role of the radiofrequency in the palliative treatment of the hepatic metastases by advanced gastric cancer, difficulty treated surgically. PMID- 16035671 TI - [Laparoscopic cholecystectomy in day-surgery analysis of feasibility in 166 consecutive patients]. AB - BACKGROUND: Day-Surgery (DS) is a widely spreading reality, both for clinical advantages to patients and organizational and economic profit to hospitals. In the last years, DS has been proposed for the treatment of a large number of diseases as inguinal and crural hernias, varicose vein, benign anorectal and thyroid pathologies. Recently, also laparoscopic cholecystectomy (LC) has been realised as DS procedure, and the initial results are promising. OBJECTIVES: To determine both the clinical feasibility of LC in DS and the factors that can predict an extension of hospital stay. MATERIALS AND METHODS: The present study is a retrospective analysis of 166 patients who underwent surgery because of symptomatic cholelithiasis in a three-year period; all patients underwent elective LC in ordinary hospital-stay. In order to identifying the patients potentially eligible for LC in DS, we carried on a selection by means of 3 consecutive stages: stage A, selection on the basis of preoperative data, stage B, selection on the basis of intraoperative factors and stage C, selection on the basis of postoperative parameters. RESULTS: Out of 166 patients, only 33 (19,8%) would have been successfully treated in DS. CONCLUSIONS: This study showed that LC in DS can be realised in strictly selected patients, on the basis of rigorous clinical and organizational criteria; furthermore, a continuous training of surgeons and nursing staff, the implementation of tele-medical facilities and the improvement of anesthesiological techniques will allow to the best results. PMID- 16035672 TI - [Mini-laparoscopic cholecystectomy: indications, technique and results]. AB - The improvement of results and patients quality life is the aim of surgical technique. Mini-laparoscopic cholecystectomy brings not only to a better cosmetic results, but also to a decrease of post operative pain, analgesic use, hospital stay and an early return to normal activities. In this study, Authors report their own experience about mini-laparoscopic cholecystectomy using 5 mm and 3 mm trocar. Patients suffering from biliary sludge, microscopic lithiasis and mild or moderate gallbladder inflammation can undergo this procedure. PMID- 16035674 TI - A new approach to the cure of the Ogilvie's syndrome. AB - The Authors describe a their own observation of 25 cases of acute colonic pseudo obstruction, better known as "Ogilvie Syndrome" with the objective to demonstrate that an early recognition and prompt appropriate therapy, better if conservative, can reduce the morbidity and the mortality of the Syndrome. The surgical therapy is reserved only to that cases in which the risk of perforation of the cecum represent an absolute indication to intervention. PMID- 16035673 TI - [Surgical treatment of abdominal trauma in pediatric age]. AB - AIM OF THE STUDY: to value the appropriateness and the efficacy of non-operative treatment in children with blunt abdominal trauma. PATIENTS AND METHODS: in this research 14 children with abdominal trauma, secondary mostly to road accidents, were studied; 9 of these had single organ injury while 5 had multiple organ injuries. Spleen has been the most injured organ (9 children), followed by liver (5 cases) and kidney (2 cases). Five children were admitted at emergency department in hypovolemic conditions, promptly corrected by resuscitative measures. All patients underwent abdominal ultrasound and/or C.T. scans in order to detect any intraperitoneal free fluid or organic injuries. Six children (43%) were followed by non-operative treatment, while other eight underwent surgery. RESULTS: all children, those treated conservatively as well as those operated, were cured, without any important complication. DISCUSSION AND CONCLUSIONS: nowadays, with the great help of ultrasound and C.T. scans, is possible to treat 40-50% of children affected by abdominal trauma with non-operative measures, with return to normal functions by the injured organs. The surgical approach is today accomplished only in presence of severe injuries or haemodynamic instability resistant to resuscitative treatment. The majority of Authors in the literature agree on the usefulness of non-operative treatment, especially for children. Regarding surgical treatment, in the near future probably we'll see a larger application of laparoscopic techniques also in the field of abdominal traumatology. PMID- 16035675 TI - [Stapled hemorrhoidectomy: critical observation on state of art]. AB - The authors review pros and cons of stapled hemorrhoidectomy (SH). Postoperative primary lower than after hemorrhoidectomy, but no data are available on the long term recurrences in large prospective series. Severe postoperative complications have been reported and SH seems less effective in patients with 4th degree files. SH is a useful technique, provided that proper indication are strictly followed and the procedure is carried out by specialists. PMID- 16035676 TI - [Silicone occlusive sheeting vs silicone cushion for the treatment of hypertrophic and keloid scars. A prospective-randomized study]. AB - BACKGROUND: Silicone gel and silicone occlusive sheeting are widely used at present for the treatment of hypertrophic and keloid scars. In recent studies the possibility was raised that static electricity generated by friction activated silicone sheeting could be the reason for this effect, and that it can, with time, cause involution of hypertrophic and keloid scars. Objective of this study was to test this hypothesis and to observe weather a continuous and also an increased negatively charged static electric field will shorten the treatment period. A silicone cushion was developed with the purpose of increasing a negative static-electric charge to accelerate the regression process. METHODS: From November 2001 to June 2002 we studied in a prospective randomized study, 72 patients with hypertrophic an keloid scars. The trial extended over a 8-month period. 37 patients underwent silicone occlusive sheeting, the remaining 35 patients underwent silicone cushion (Clinicel). RESULTS: Treatment with the silicone cushions yielded 74,2% cessation of itching and burning followed by pallor and flattening of the scar, some markedly so, over a few weeks to 5 months period. Additional 25,7% had their scars resolved in up to 8 months of treatment. Four patients (11,4%) who add recalcitrant scars with little response to the use of the silicone cushion were given intralesional corticosteroid injections, in addiction to the use of the cushion, resulting in a fairly rayed resolution of these scars over a period of 2 months. Treatment with the silicone occlusive sheeting yielded 52,3% itching and burning cessation followed by pallor and flattening of the scar, some markedly so, over a few weeks to 5 months period. Additional 22,1% had their scars resolved in up to 8 months of treatment. In conclusion by comparing the results of this trial using silicone cushions for the treatment of hypertrophic and keloid scars with those obtained using silicone gel or occlusive sheeting, a much faster response was demonstrated. PMID- 16035677 TI - Partial acute thrombosis of internal carotid artery: a case report. AB - OBJECTIVE: The effectiveness of the urgent carotid TEA in patients with not stabilizes serious neurological symptoms, remains controversial. MATERIAL: The authors report on a case of a partial acute carotid thrombosis. The patient had outcomes of pulmonary tumor dealt with surgical removal of the medium right lobe. At admission, the neurological examination evidenced non specific neurological deficits to both the advanced limbs above all. The blood test evidenced 628000 platelets. The color-doppler showed in the internal carotid artery of right the presence of floating thrombus, during very small fibro-calcified plate, likely ulcerated. RESULTS: The patient was submitted to surgical urgent procedure without ulterior diagnostic deepening. Such attitude was justified from the total regression of the symptomatology and from the absence of alterations of the BEE. DISCUSSION: The urgent surgical treatment has had good outcome. In literature there are similar cases but not place upon ones because of "aetiology" to the inner presence of floating thrombus in carotid not completely occluding in all and the three cases, causes do not correspond from alters platelet parameters, but thrombus was of cardiogenic origin in one case, it was caused by hormonal therapy for sterility in the second, there was thrombosis of carotid aneurysm in third. CONCLUSION: We remark the importance of eco-color Doppler for urgent diagnosis of acute thrombosis, and timeliness of surgical treatment in the "acute" patient with attenuation or regression of the debut symptomatology. PMID- 16035678 TI - Esophacoil for palliation of recurrent malignant esophago-jejunal anastomotic stricture. Case reports. AB - Recurrent malignancy at esophagojejunal anastomosis represents an incurable stage of disease. Treatment options are limited. Surgery presents a high rate of morbidity and mortality and frequently fails to alleviate symptoms. Radiochemotherapy relieves dysphagia slowly and offers disappointing clinical results. The use of Self Expanding Metallic Stent (SEMS) has shown to be effective, safe and offers a good quality of residual life. We herein report three cases of malignant esophagojejunal anastomotic strictures successfully treated with metallic coil stent (Esophacoil). PMID- 16035679 TI - Heterotopic gastric mucosa in the gallbladder: case report and literature review. AB - INTRODUCTION: we report on a case of heterotopic gastric mucosa in the neck of the gallbladder and we also review 95 other reports of HGM in the gallbladder in the international medical literature from 1977. AIM: to evaluate the gold standard treatment in heterotopic gastric mucosa of the gallbladder by the analysis of literature, compared with our anecdotal experience. PATIENT AND METHOD: a 43-year-old man, who was recently symptomatic, visited our hospital to submit to laparoscopic cholecistectomy for cholelithiasis. Ultrasonography revealed a broad-based polypoid lesion in the gallbladder (2.5 cm in diameter in the neck of the gallbladder), with multiple gallstones. RESULTS: standard laparoscopic cholecystectomy was performed. The specimen revealed a 2.5 x 1.7 x 0.5 cm polypoid lesion with deep in the body, with many gallstones in the gallbladder. Histologically, the polypoid lesion consisted of gastric fundic glands located only in the mucosa of the gallbladder. The surrounding mucosa consisted of almost normal epithelium without any metaplastic changes. Postoperative technetium 99m-pertechnetate scintigraphy demonstrated no evidence of gastric heterotopia elsewhere in the body. Actually the patient is in long time follow-up, asymptomatic. CONCLUSIONS: for its extreme difficult to make a conclusive diagnosis and thereby rule out the possibility of cancer, it appears that laparoscopic cholecystectomy may be unavoidable for patients affected by heterotopic gastric mucosa at the present time and care must be taken when a diagnosis is made based on intraoperative frozen sections. PMID- 16035680 TI - Genetic testing: when to test, when to refer. PMID- 16035681 TI - Long-term standard-dose warfarin to prevent thrombosis. PMID- 16035682 TI - Imiquimod to treat basal cell and squamous cell carcinomas. PMID- 16035683 TI - Low-molecular-weight heparin for initial treatment of venous thromboembolism. PMID- 16035684 TI - Antibiotics for acute laryngitis in adults. PMID- 16035685 TI - Follow-up after surgically treated breast cancer. PMID- 16035686 TI - Prevention of falls in older patients. AB - Falls are one of the most common geriatric syndromes threatening the independence of older persons. Between 30 and 40 percent of community-dwelling adults older than 65 years fall each year, and the rates are higher for nursing home residents. Falls are associated with increased morbidity, mortality, and nursing home placement. Most falls have multiple causes. Risk factors for falls include muscle weakness, a history of falls, use of four or more prescription medications, use of an assistive device, arthritis, depression, age older than 80 years, and impairments in gait, balance, cognition, vision, and activities of daily living. Physicians caring for older patients should ask about any falls that have occurred in the past year. Assessment should include evaluating the circumstances of the fall and a complete history and physical examination, looking for potential risk factors. The most effective fall prevention strategies are multifactorial interventions targeting identified risk factors, exercises for muscle strengthening combined with balance training, and withdrawal of psychotropic medication. Home hazard assessment and modification by a health professional also is helpful. PMID- 16035687 TI - Information from your family doctor. How to prevent falling. PMID- 16035688 TI - Diabetic nephropathy: common questions. AB - Diabetic nephropathy, or diabetic kidney disease, affects 20 to 30 percent of patients with diabetes. It is a common cause of kidney failure. Diabetic nephropathy presents in its earliest stage with low levels of albumin (microalbuminuria) in the urine. The most practical method of screening for microalbuminuria is to assess the albumin-to-creatinine ratio with a spot urine test. Results of two of three tests for microalbuminuria should be more than 30 mg per day or 20 mcg per minute in a three- to six-month period to diagnose a patient with diabetic nephropathy. Slowing the progression of diabetic nephropathy can be achieved by optimizing blood pressure (130/80 mm Hg or less) and glycemic control, and by prescribing an angiotensin-converting enzyme inhibitor or angiotensin receptor blocker. Patients with diabetes and isolated microalbuminuria or hypertension benefit from angiotensin-converting enzyme inhibitors or angiotensin receptor blockers. In the event that these medications cannot be prescribed, a nondihydropyridine calcium channel blocker may be considered. Serum creatinine and potassium levels should be monitored carefully for patients receiving angiotensin-converting enzyme inhibitors or angiotensin receptor blockers. These medications should be stopped if hyperkalemia is pronounced. PMID- 16035689 TI - Information from your family doctor. Diabetic nephropathy. PMID- 16035690 TI - Health effects of garlic. AB - Garlic has long been used medicinally, most recently for its cardiovascular, antineoplastic, and antimicrobial properties. Sulfur compounds, including allicin, appear to be the active components in the root bulb of the garlic plant. Studies show significant but modest lipid-lowering effects and antiplatelet activity. Significant blood pressure reduction is not consistently noted. There is some evidence for antineoplastic activity and insufficient evidence for clinical antimicrobial activity. Side effects generally are mild and uncommon. Garlic appears to have no effect on drug metabolism, but patients taking anticoagulants should be cautious. It seems prudent to stop taking high dosages of garlic seven to 10 days before surgery because garlic can prolong bleeding time. PMID- 16035691 TI - Diagnosis and management of fragile X syndrome. AB - To complement the 2005 Annual Clinical Focus on medical genomics, AFP will be publishing a series of short reviews on genetic syndromes. This series was designed to increase awareness of these diseases so that family physicians can recognize and diagnose children with these disorders and understand the kind of care they might require in the future. The first review in this series discusses fragile X syndrome. PMID- 16035693 TI - Raloxifene for prevention of osteoporotic fractures. PMID- 16035692 TI - Diagnosis of acute coronary syndrome. AB - The term "acute coronary syndrome" encompasses a range of thrombotic coronary artery diseases, including unstable angina and both ST-segment elevation and non ST-segment elevation myocardial infarction. Diagnosis requires an electrocardiogram and a careful review for signs and symptoms of cardiac ischemia. In acute coronary syndrome, common electrocardiographic abnormalities include T-wave tenting or inversion, ST-segment elevation or depression (including J-point elevation in multiple leads), and pathologic Q waves. Risk stratification allows appropriate referral of patients to a chest pain center or emergency department, where cardiac enzyme levels can be assessed. Most high-risk patients should be hospitalized. Intermediate-risk patients should undergo a structured evaluation, often in a chest pain unit. Many low-risk patients can be discharged with appropriate follow-up. Troponin T or I generally is the most sensitive determinant of acute coronary syndrome, although the MB isoenzyme of creatine kinase also is used. Early markers of acute ischemia include myoglobin and creatine kinase-MB subforms (or isoforms), when available. In the future, advanced diagnostic modalities, such as myocardial perfusion imaging, may have a role in reducing unnecessary hospitalizations. PMID- 16035694 TI - Screening for genital herpes. PMID- 16035695 TI - Multiple erythematous plaques of the trunk. PMID- 16035696 TI - Extended prophylaxis of venous thromboembolism in major orthopaedic surgery. AB - The risk of postoperative venous thromboembolism continues after discharge from the hospital. Studies in patients undergoing hip replacement or hip fracture surgery consistently found the rate of asymptomatic deep vein thrombosis to be substantially reduced by extended out-of-hospital prophylaxis and meta-analyses demonstrate symptomatic deep vein thrombosis to be reduced in parallel with asymptomatic. On the basis of these data, extended prophylaxis is recommended in hip replacement and hip fractures. The recommendation is particularly strong for patients with additional personal risk factors. PMID- 16035697 TI - Review of methods to quantify lag screw placement in hip fracture fixation. AB - This review considers methods used to quantify lag screw placement within the femoral head following proximal femoral fracture. Implants used for fixation of femoral neck fractures can lead to unwanted outcomes in some patients. Cutting out of the screw within the femoral head has been shown to be the commonest cause of failure and is related to lag screw placement within the femur. Until now, there have been two methods published which quantify lag screw position within the femoral head. These are the concepts of 'Tip Apex Distance' and Parker's ratio method of lag screw placement. We shall discuss each one in turn and their implications in terms of fracture fixation failure. PMID- 16035698 TI - Surgical treatment of sprengel's deformity: a modified Green procedure. AB - Twelve cases of Sprengel's deformity were treated surgically by a modification of Green's procedure. After clavicular osteotomy, all muscular attachments to the scapula were freed; an omovertebral band if present was cut and the scapula was rotated and moved caudal to a more normal position, to be sutured into a pocket of the latissimus dorsi. The patients included 10 female and 2 male patients (age range at the time of operation: 3 to 9 years; mean: 5.2 years). The deformity involved the left shoulder in 8 patients, the right shoulder in 4. All patients were followed for an average of 2 years (range: 6 months to 4.5 years). The range of flexion improved from an average of 95 degrees to 105 degrees and the range of abduction from an average of 81 degrees to 97 degrees. Cosmesis was improved in all 12 patients but three patients had an unsightly surgical scar. Surgical treatment by a modified Green procedure for patients with Sprengel's deformity is indicated when the patient and family want cosmetic or functional improvement. PMID- 16035699 TI - Suprascapular neuropathy in volleyball players. AB - Entrapment neuropathy of the suprascapular nerve (SNE), although a recognised clinical entity, is a relatively rare cause of shoulder pain and subjective weakness in the athlete involved in overhead sports like volleyball and badminton. This study deals with the presentation and management of four unusual cases of suprascapular nerve entrapment in volleyball players. Four male volleyball players presented to our department with intractable shoulder pain and subjective sensation of shoulder weakness. They all had visible wasting of both supraspinatus and infraspinatus muscles, together with weakness of abduction and external rotation of the arm. They all responded temporarily to a diagnostic injection of local anaesthetic. MR imaging was useful in diagnosing space occupying lesions in three cases and the presence of a hypertrophic suprascapular ligament in one case. Due to failure of non- operative treatment, which included activity modification, rest, analgesics and rehabilitation programme over 6 months, surgery was then required to decompress the suprascapular nerve. All patients were symptom free at 6 months postoperatively and after an intensive rehabilitation programme, they were able to return to their normal level of activity including sport. PMID- 16035700 TI - Results with Kudo elbow prostheses in non-traumatic indications : a study of 36 cases. AB - With a mean follow-up of 62 months, we reviewed 13 Kudo type 4 and 23 Kudo type 5 elbow prostheses implanted for non-traumatic indications in 30 patients. Clinical results were assessed with the Mayo Clinic Performance Index (MCPI) taking into account pain, mobility, stability and daily activities. Postoperatively, pain disappeared or decreased, ulnar nerve dysfunction was improved and the functional status was significantly improved. The average range of motion increased by 7.8 degrees in extension and by 11.5 degrees in flexion; pronation and supination remained unchanged. Clinically, 89% of patients scored poorly on the MCPI preoperatively, whereas 61% had excellent or good scores postoperatively. Twenty two patients out of 30 were satisfied. There were two early prosthetic dislocations, and prosthetic instability, assessed clinically, was found in 6 cases (17%). The rate of loosening was 28% (10/36) and seven elbows were revised. Metallosis, loosening and instability were significantly correlated. The survival rate of 82% at 54 months (SD = 7) is lower than that reported for non-constrained and semi-constrained prostheses in the literature. PMID- 16035701 TI - Disability and function after arthroscopic repair of ulnar avulsions of the triangular fibrocartilage complex of the wrist. AB - Ulnar wrist pain due to a lesion of the triangular fibrocartilage complex (TFCC) is frequent. Based on studies of the vascularity, (traumatic) ulnar avulsions can be sutured. Arthroscopic techniques have been designed but results are scarcely published. This is a follow-up study of 52 patients with an ulnar avulsion of the TFCC. All patients were treated with an arthroscopic technique. Evaluation was directed to the subjective outcome, functionality ( using the DASH score) and objective parameters. The mean DASH score was 17.3. Pain was absent or minimal in 47 patients. The grip force was 80% of the contralateral side. Based on these findings, arthroscopic repair of the TFCC appears to be a reliable technique. PMID- 16035702 TI - Unsatisfactory results with the cementless Omnifit acetabular component due to polyethylene and severe osteolysis. AB - A high incidence of acetabular osteolysis (43%), associated with osteolytic lesions in the proximal femur (22.6%) and leading to a high revision rate, was experienced with the Omnifit total hip prosthesis. We reviewed the clinical and radiological results with 429 Omnifit total hips in 356 patients after a mean follow-up of 60 months. Time to revision and wear of the polyethylene liner with different acetabular shell types were specifically analysed. Pelvic osteolysis first became manifest in the acetabular bone opposite to the holes in the metal shell. Osteolysis occurred predominantly adjacent to the central hole in the metal shell of threaded cups; widespread and larger defects were found in press fit cups with peripheral screw holes. Kaplan Meier survival analysis demonstrated a higher probability for retaining the threaded cup at 6 years (96%; 95% confidence interval: 93-99%) compared to the survival of the press fit cup (66%; 95%-CI: 56-77%). The results suggest a negative relationship between backside wear, the larger number of holes in the cup, the extent of osteolysis and survival rate of the press fit cups. Based on these findings and supported by similar reports about osteolysis related to the same cup design, it was hypothesised that backside wear due to the insufficient locking mechanism of the Omnifit acetabular cup was the major cause of the unsatisfactory results in our patients. For this reason we discontinued using this type of uncemented socket. PMID- 16035703 TI - Reconstruction of the medial patellofemoral ligament for patellar instability using a semitendinosus autograft. AB - Between 1999 and 2001 thirty knees underwent a semitendinosus tendon plasty to recreate the medial patellofemoral ligament for recurrent patellar dislocation. The mean follow-up was 38 months. The mean improvement of the patellofemoral congruence angle after surgery was 14 +/- 7 degrees. All patients ended up with a full range of motion, except one patient, whose flexion was limited to 120 degrees due to superficial wound infections. Dislocation did not recur. According to the Larsen and Lauridsen outcome score the clinical results were excellent in 27 patients, good in 2 and fair in one. In conclusion this procedure is indicated for the chronic dislocation and cases of severe femoral dysplasia with marked laxity. The procedure assures the stabilisation of the patella, although it doesn't restore the patellofemoral congruence angle to normal values. PMID- 16035704 TI - Day case anterior cruciate ligament reconstruction: a study of 51 consecutive patients. AB - We prospectively assessed 51 arthroscopy assisted anterior cruciate reconstructions done over two years as day cases. We looked at the clinical results, postoperative pain control and patient satisfaction. There were 45 males and 6 females, with ages ranging from 18 to 52 years. A hamstring graft was used in 38 patients while 13 patients had reconstructions using the patellar tendon. Forty-nine patients were successfully discharged on the same day while 2 patients had to be admitted due to excessive drainage. Six patients had additional procedures at the time of surgery. The mean time interval from injury to surgery was 27 months (range: 2 to 180). Forty-six patients had an excellent to good outcome as regards satisfaction with one poor result. Pain control was not a major issue with any of the other patients included in the study. Our study demonstrates that day case arthroscopic ACL reconstruction is a safe procedure with minimal to absent morbidity. PMID- 16035705 TI - Two-stage total knee arthroplasty for non-salvageable septic arthritis in diabetes mellitus patients. AB - Pyogenic knee arthritis in a patient with advanced osteoarthritis is a serious medical problem. We have performed arthroscopic debridement in 136 patients with pyogenic knee arthritis from January 1999 to December 2001. Five of these patients were diabetic, they did not respond to the standard treatment protocol and they continued to have infection. For these patients, we performed open arthrotomy, with implantation of antibiotic cement as a spacer, and staged total knee arthroplasty. The clinical results were evaluated using the Hospital for Special Surgery (HSS) scoring system. At an average follow-up of 38 months (range: 29 to 46), the average pain score was 83 and the functional score was 73 with no patient having recurrence of the infection. This study shows that just as a 2-stage revision is now done for infected total knee arthroplasty, primary uncontrolled infected knees may be treated by a 2-stage arthroplasty as well. PMID- 16035706 TI - Lengthening procedures of small bones of foot and foot stump. AB - Foot length discrepancy may result from congenital or acquired causes. If the absence of the foot is more proximal than the metatarsal level, push off and foot resilience will be disturbed and rapid walking and spring will be awkward. Those patients have to be fitted with a prosthesis extending above the ankle to the distal leg. The functional impairment and poor cosmetic appearance become social problems especially for adolescents. Twelve cases underwent a lengthening procedure of small bones of the foot in our clinic since 1995 to lengthen the foot or a foot stump. Results were satisfactory. PMID- 16035707 TI - Idiopathic multicentric osteolysis: a case report and literature review. PMID- 16035708 TI - Treatment of high-grade spondylolisthesis by posterior lumbosacral transfixation with transdiscal screws: surgical technique and preliminary results in four cases. AB - In high-grade spondylolisthesis, the surgical treatment should be aimed at achieving good stability to allow solid fusion in the face of high biomechanical forces at the lumbosacral junction. A 360 degrees fusion seems to be able to provide this stability. This is however extensive surgery and many problems and complications have been reported. In order to overcome these difficulties, various new procedures have been published. Most of these techniques aim for a good anterior column support, allowing primary stability and a large bony surface area for fusion. Transfixation of the lumbosacral disc space using a fibular strut graft was published decades ago. Several modifications have been reported since, including the use of threaded cages filled with bone graft. In contrast to the number of these surgical techniques, only few biomechanical test results and small-size clinical studies have been reported in the literature. An interesting technique of lumbosacral transfixation includes the use of transdiscal pedicle screws, described by Abdu et al in 1994. This allows for the use of standard instruments and implants, while biomechanical testing recently has shown improved stability equal to classic PLIF constructs by providing three-column support. Moreover, in high-grade slips this technique is easier to perform than other methods. We have treated four consecutive patients according to this technique with good clinical and radiographic results. The surgical technique is described in detail and a review of the literature is provided. PMID- 16035709 TI - Influence of burst TENS stimulation on collagen formation after Achilles tendon suture in man. A histological evaluation with Movat's pentachrome stain. AB - Retrograde or antidromic stimulation of the nociceptive C fibres is known to lead to the release of sensory neuropeptides, such as substance P (SP), by the peripheral endings of sensory unmyelinated C nerve fibres. These neuropeptides play a role in the healing of soft tissues. Burst TENS (Transcutaneous Electric Nerve Stimulation) is known to be most effective in influencing retrograde C fibre-evoked activity. This is why burst TENS was used in a randomised study as a stimulus for the healing of the sutured Achilles tendon in 9 patients, versus 9 others who received no stimulus. Originally, each group consisted of 10 patients, but there was a single drop-out in each group. Six weeks after surgery a needle biopsy sample was obtained, and stained with Movat's pentachrome stain. It showed a statistically significant influence of burst TENS on new collagen production, maturation of newly formed collagen and organisation of collagen. This suggests that burst TENS might positively influence healing of Achilles tendon suture in man. PMID- 16035710 TI - External fixation of finger fractures made simple. AB - The authors present a simple method for treatment of comminuted and compound phalangeal fractures in the hand, using readily available K-wires and needle sheaths. PMID- 16035711 TI - Posterior stabilisation of a malignant cervico-thoracic vertebral bone defect. AB - Oesophageal cancer is frequently complicated by malignant fistulae. Necrosis of the tumour following radiotherapy or chemotherapy may lead to the development of fistulae between the oesophagus and adjacent tissues and organs. We report the expansion of an extra-luminal oesophageal cancer after resection, invading the cervico-thoracic spine, fortunately without neurological deficit, and leading to instability and formation of a malignant fistula linking the tracheo-bronchial tree to the subarachnoidal space. To prevent imminent paraplegia and to alleviate severe pain, we rigidly stabilised the spine at the cervico-thoracic junction using an angle-stable system through a single posterior approach. Further postoperative follow-up revealed no signs of neurological deterioration. Cervico thoracic stability was preserved until the patient died nearly five months postoperatively. This case shows that posterior stabilisation and decompression may be a palliative option for patients with imminent paraplegia and severe pain due to advanced tumour infiltration of the cervico-thoracic spine. PMID- 16035712 TI - Divergent dislocation of the ring and little finger carpometacarpal joints--a rare injury pattern. AB - Hand injuries due to longitudinal forces in the line of the metacarpals demonstrate unusual dislocation patterns. We describe a case of volar intra articular fracture dislocation of the ring finger carpometacarpal joint in association with a pure dorsal dislocation of the little finger carpometacarpal joint. Open reduction supplemented with Kirschner wire fixation restored normal carpometacarpal joint anatomical relations and achieved an excellent clinical result. PMID- 16035713 TI - Unusual case of thoracic outlet syndrome caused by a neurilemmoma in the pectoralis minor space. AB - A 34-year-old man presented with a 5-year history of paraesthesia of the right palm and the right middle and ring fingers. This paraesthesia was exacerbated by elevation of the right arm. A tumour was palpable in the subclavicular fossa. As magnetic resonance imaging (MRI) indicated a neurogenic tumour originating from the brachial plexus, a diagnosis of thoracic outlet syndrome caused by a neurilemmoma in the pectoralis minor space was made. Pathological examination showed the tumour to be a benign neurilemmoma. There have been only three previous case reports of neurilemmomas as causes of thoracic outlet syndrome worldwide, and this is the first report of a neurilemmoma originating from the lateral fascicles of the brachial plexus in the pectoralis minor space causing thoracic outlet syndrome. PMID- 16035714 TI - Pelvic girdle sepsis in childhood. An illustrative case of the difficulty in diagnosis. AB - The child who presents with fever, limp and hip pain will often undergo multiple diagnostic procedures before a definitive diagnosis is made. We describe a diagnostically challenging case of a 14-year-old boy presenting with an atraumatic painful limp and pyrexia. Eventually the diagnosis of obturator internus muscle abscess with associated ischial osteomyelitis was made. Of the 19 previous cases reported, four children had associated osteomyelitis and were of an older age. Symptomatology varies, clinical examination is non-specific and the diagnosis can be difficult. Haematological indices are more predictive than in cases of classical osteomyelitis or septic arthritis. Subtle features on conventional radiography and isotope bone scanning should not be overlooked while CT and MRI may be complementary in diagnosis. PMID- 16035715 TI - Tuberculosis of the ilium: is it really so rare? AB - Tuberculosis of the ilium is a rare identity, accounting for less than 1% of all skeletal tuberculosis. We report two such lesions in immunocompetent individuals. Tuberculosis remains an important differential diagnosis when faced with unusual or chronic bony lesions, especially in endemic areas, even in non immunocompromised individuals. It can involve any site and affect people of any age. PMID- 16035716 TI - Superior dislocation of the patella. Case report and review of literature. AB - Superior dislocation of the patella is a rare diagnosis, which usually occurs after a trivial trauma. It usually requires manipulation with analgesics or may even need anaesthesia. We report a case of spontaneous reduction of the dislocation, which lead us to believe that this may be more common in the community than has been reported. PMID- 16035717 TI - Recalcitrant scoliosis in Proteus syndrome. AB - Proteus syndrome is a rare congenital condition with various clinical features such as hemihypertrophy, macrodactyly, subcutaneous masses, brain-like hyperplasia of the soles and/or palms, epidermal naevi and scoliosis with other mesodermal malformations. Multifocal overgrowth can affect various tissues causing severe functional and cosmetic disability, but intellectual and language development are mostly normal. Orthopaedic problems include macrodactyly, hindfoot deformity, limb length inequality, genu valgum and scoliosis. Usually, scoliosis does not respond to bracing, and surgical intervention may be required. Despite surgical correction, instrumentation and fusion, progression of deformity can occur. The authors describe a case with a Th7-L2 scoliosis, which completely relapsed, 20 months after posterior instrumented fusion. Surgery should not be undertaken lightly, given the abnormal growth potency typical for Proteus syndrome. PMID- 16035719 TI - Fair trade in care? PMID- 16035718 TI - Hearing voices? Don't be afraid. PMID- 16035720 TI - We don't do that. PMID- 16035721 TI - What older people want. PMID- 16035722 TI - Ask the experts. Assistive technology. PMID- 16035723 TI - Respite care and short-break support: new forms of an old idea? PMID- 16035724 TI - Primary healthcare teams and dementia. PMID- 16035725 TI - Arthritis in the older person: Part 2. AB - Part 1 of this article, in last month's Nursing Older People, identified the major musculoskeletal disorders affecting older people, the signs and symptoms of these and drug therapies available. Part 2 discusses non-pharmacological treatment options, patient education, multi-disciplinary support and how people living with musculoskeletal disorders can be helped to achieve quality of life PMID- 16035726 TI - Recognising acute deterioration. PMID- 16035727 TI - A privilege that pays. PMID- 16035728 TI - A big hand for almshouses. PMID- 16035729 TI - Heart-fatty acid-binding protein as a marker for early detection of acute myocardial infarction and stroke. AB - Heart-fatty acid-binding protein (H-FABP) is a small cytosolic protein involved in intracellular fatty acid transport. This protein, highly concentrated in the heart, is quickly released into plasma after myocardial injury. Results from numerous studies suggest that H-FABP is an excellent marker for the early detection of myocardial damage. H-FABP is also expressed in the brain, although in lower concentrations than in the heart. Recent preliminary studies also investigated the usefulness of H-FABP for the diagnosis of acute and chronic neurological disorders. These potential applications of H-FABP in clinical practice are reviewed in this article, with a strong focus on the early diagnosis of acute myocardial infarction and stroke. PMID- 16035730 TI - Paraoxonase 1-192Q allele is a risk factor for idiopathic chronic pancreatitis. AB - BACKGROUND: The cause of chronic pancreatitis (CP) remains unknown. However, oxidative stress might play a role since recent animal studies have demonstrated that oxygen-free radicals contribute to the pathogenesis of experimental pancreatitis. Human serum paraoxonase (PON1) is an antioxidant enzyme that protects against cellular damage from oxidative stress. Genetic variations resulting in variable activity rates of this enzyme, are of toxicological and physiological importance. AIM: We investigated whether genetic polymorphisms of the PON1 gene modify the risk for CP. MATERIALS AND METHODS: DNA samples were obtained from 236 adult CP patients of hereditary (n = 23), alcoholic (n = 137), or idiopathic (n = 76) origin. DNA from 113 healthy controls and from 93 alcoholic controls were analyzed for comparison. Patients and controls were all of Caucasian origin. Genetic polymorphisms (L55M and Q192R) in PON1 were determined by PCR, followed by restriction fragment length polymorphism analyses in all subjects. RESULTS: The frequencies of the PON1-55 alleles did not differ between CP patients and healthy controls. However, the PON1-192Q allele was significantly more common in idiopathic CP patients (OR : 1.5, 95% CI 1.02, 2.5) compared with healthy controls. CONCLUSIONS: These data suggest that the PON1 192Q allele, resulting in partly deficient antioxidant and detoxification activity of this enzyme, might be a risk factor for idiopathic CP in Caucasians. PMID- 16035731 TI - Molecular analysis of the p27/kip1 gene in breast cancer. AB - BACKGROUND: Genetic polymorphisms and mutations of the genes involved in tumorigenesis may determine individual susceptibility for cancer. The p27/Kip1 protein belongs to the family of cyclin-dependent kinase-inhibitory proteins, which are negative regulators of cell-cycle progression. Reduced protein levels of p27/Kip1 have been reported in numerous human cancers including breast cancer. METHODS AND RESULTS: p27 gene mutations and the codon 109 polymorphism were investigated in breast cancer patients by single strand conformation polymorphism analysis, PCR-restriction fragment length polymorphism analysis and DNA sequencing. Mutations were identified in 2 of 24 breast tumor samples. One G-->A transition resulting in a silent mutation and a single base deletion resulting in a nonsense mutation were detected in one patient. Another breast cancer sample harbored a T-->A transition at codon 159. An association between the codon 109 B allele and breast cancer was observed. CONCLUSION: Our study indicates that mutational alterations in the p27 gene are rare in human breast cancer. The codon 109 B allele is associated with high-grade tumors. PMID- 16035732 TI - Touch-down reverse transcriptase-PCR detection of IgV(H) rearrangement and Sybr Green-based real-time RT-PCR quantitation of minimal residual disease in patients with chronic lymphocytic leukemia. AB - BACKGROUND: Patients with chronic lymphocytic leukemia (CLL) can relapse even after aggressive therapy and autografts. It is commonly assumed that to prevent relapse the level of minimal residual disease (MRD) should be as low as possible. To evaluate MRD, highly sensitive quantitative assays are needed. AIM: The aim of the study was to develop a robust and sensitive method for detection of the clonal immunoglobulin heavy-chain variable (IgV(H)) rearrangement in CLL and to introduce a highly sensitive and specific methodology for MRD monitoring in patients with CLL who undergo intensive treatment. METHODS: As a prerequisite for MRD detection, touch-down reverse transcriptase (RT)-PCR using degenerate primers were used for the diagnostic identification of (H) gene rearrangement(s). For quantitative MRD detection in 18 patients, we employed a real-time RT-PCR assay (RQ-PCR) making use of patient-specific primers and the cost-saving Sybr-Green reporter dye (SG). For precise calibration of RQ-PCR, patient-specific IgV(H) sequences were cloned. RESULTS: Touch-down RT-PCR with degenerate primers allowed the successful detection of IgV(H) clonal rearrangement(s) in 252 of 257 (98.1%) diagnostic samples. Biallelic rearrangements were found in 27 of 252 (10.7%) cases. Degenerate primers used for the identification of clonal expansion at diagnosis were not sensitive enough for MRD detection. In contrast, our RQ-PCR assay using patient-specific primers and SG reached the sensitivity of 10(-)(6). We demonstrated MRD in each patient tested, including four of four patients in complete remission following autologous hematopoietic stem cell transplantation (HSCT) and three of three following allogeneic 'mini'-HSCT. Increments in MRD might herald relapse; aggressive chemotherapy could induce molecular remission. CONCLUSIONS: Our touch-down RT-PCR has higher efficiency to detect clonal IgV(H) rearrangements including the biallelic ones. MRD quantitation of IgV(H) expression using SG-based RQ-PCR represents a highly specific, sensitive, and economic alternative to the current quantitative methods. PMID- 16035733 TI - Instability of Trypanosoma cruzi DNA in blood lysates: importance for PCR DNA based diagnosis. AB - OBJECTIVE: In order to evaluate the stability of Trypanosoma cruzi kinetoplast DNA (kDNA), the blood samples from seven patients with Chagas disease were stored in different buffers and at different temperatures. METHODS: Three different buffers were used: buffer A, 6 mol/L guanidine-HCl; buffer B, 6M guanidine-HCl and 0.2M EDTA pH 7.5; and buffer C, 6M guanidine-HCl, 0.2M EDTA pH 7.5 and 10 microM dl-alpha-tocopherol (Roche, Basal, Switzerland). Two temperatures were used: 4 degrees C and 25 degrees C. Vitamin E was added to the blood lysates as an antioxidant. T. cruzi kDNA was obtained by phenol extraction, and then PCR amplifications and Southern blot were carried out in each DNA sample up to 90 days of blood storage. The iron content of each sample was determined by atomic absorption spectrophotometry. RESULTS: Overall, there is an association between T. cruzi kDNA stability and the storage time of blood samples. No significant differences were detected in T. cruzi kDNA stability in the presence or absence of vitamin E or with citrate or EDTA as an anticoagulant. There was no statistical difference in the failure of PCR-based kDNA detection with these different storage buffers, temperatures or iron levels. CONCLUSIONS: The blood lysates promote T. cruzi kDNA damage in a time-dependent manner that reduces the ability to detect the genomic DNA of an infectious agent by PCR. The high concentration of guanidine-HCl denatured proteins in these storage conditions probably denotes a non-enzymatic kDNA lysis. PMID- 16035736 TI - Reaction of SO2 with AuCeO2(111): importance of O vacancies in the activation of gold. AB - Synchrotron-based high-resolution photoemission was used to study the adsorption and chemistry of SO(2) on AuCeO(2)(111) and AuO(x)CeO(2) surfaces. The heat of adsorption of the molecule on Au nanoparticles supported on stoichiometric CeO(2)(111) was 4-7 kcalmol larger than on Au(111). However, there was negligible dissociation of SO(2) on the AuCeO(2)(111) surfaces. The full decomposition of SO(2) was observed only after introducing O vacancies in the ceria support. AuO(x)CeO(2) surfaces were found to be much less chemically active than AuCeO(2)(111) or AuCeO(2-x)(111) surfaces. The active sites in {Au + AuO(x)}ceria catalysts should involve pure gold nanoparticles in contact with O vacancies. PMID- 16035737 TI - Observation of resonant interatomic coulombic decay in Ne clusters. AB - We have measured the electron spectra of Ne clusters after excitation with photon energies around the 2s inner valence threshold. At two photon energies below threshold, a resonantly enhanced surplus of low kinetic-energy electrons is observed. The kinetic energy of the peak does not vary with the photon energy and is slightly larger than the transition energy of Interatomic Coulombic Decay (ICD) above threshold. This leads us to assume that an ICD-like process is present. In analogy to the Auger and the resonant Auger decay this new phenomenon is termed resonant ICD. PMID- 16035738 TI - Relating kinetic rates and local energetic roughness by accelerated molecular dynamics simulations. AB - We show that our accelerated molecular-dynamics (MD) approach can extend the time scale in all-atom MD simulations of biopolymers. We also show that this technique allows for the kinetic rate information to be recaptured. In deducing the kinetic rates, the relationship between the local energetic roughness of the potential energy landscape and the effective diffusion coefficient is established. These are demonstrated on a very slow but important biomolecular process: the dynamics of cis-trans-isomerization of Ser-Pro motifs. We do not only recapture the slow kinetic rates, which is difficult in traditional MD, but also obtain the underlying roughness of the energy landscape of proteins at atomistic resolution. PMID- 16035739 TI - Different chemical dynamics for different conformers of biological molecules: photoionization of glycine. AB - Single-photon ionization dynamics of two conformers of glycine is studied by classical trajectory simulations using the semiempirical PM3 potential surface in "on the fly" calculations. Initial conditions for the trajectories are weighted according to the Wigner distribution function computed for the initial vibrational ground state. Vertical ionization in the spirit of the classical Franck-Condon principle is assumed. The dynamics of the two conformers are compared during the first 10 ps. The comparison shows very different dynamical behavior for the two conformers. In particular, the chemical fragmentation pathways differ in part. Also, one of the conformers gives much higher rates of conformational transitions, while the other conformer gives larger chemical fragmentation yields. The example shows significantly different chemical dynamics for two conformers close in energy and separated by a low barrier. PMID- 16035740 TI - An efficient approach for the calculation of Franck-Condon integrals of large molecules. AB - A general and efficient approach for the calculation of Franck-Condon integrals (FCIs) of large molecules is presented. In a first step, by exploiting the diagonally dominant and sparse structure of the Duschinsky matrix, a model system is constructed for which the Duschinsky matrix takes a block-diagonal form. For each of these blocks separately, the FCIs are calculated discarding all below a certain threshold. From those integrals retained the FCIs of the model system are obtained by simple multiplication. These serve as an estimate for the FCIs of the exact system which are calculated for those integrals which lie above a certain threshold. By systematically decreasing the threshold, the simulation can be reliably converged to the exact result with an arbitrary accuracy. Using this scheme, a considerable reduction of the number of FCIs which have to be calculated is achieved which leads to an improved scaling behavior of the computational effort with system size. The approach has been tested thoroughly for a set of molecules including difficult cases. For the larger systems a speedup of up to three orders of magnitude compared to an exact calculation is observed while the errors can be kept negligible. With this approach accurate calculations of FCIs are feasible also for large molecules encountered in "real life" chemistry, especially biochemistry and material science. PMID- 16035741 TI - Open-shell localized Hartree-Fock method based on the generalized adiabatic connection Kohn-Sham formalism for a self-consistent treatment of excited states. AB - An effective exact-exchange Kohn-Sham approach for the treatment of excited electronic states, the generalized adiabatic connection open-shell localized Hartree-Fock (GAC-OSLHF) method is presented. The GAC-OSLHF method is based on the generalized adiabatic connection Kohn-Sham formalism and therefore capable of treating excited electronic states, which are not the energetically lowest of their symmetry. The method is self-interaction free and allows for a fully self consistent computation of excited valence as well as Rydberg states. Results for atoms and small- and medium-size molecules are presented and compared to restricted open-shell Hartree-Fock (ROHF) and time-dependent density-functional results as well as to experimental data. While GAC-OSLHF and ROHF results are quite close to each other, the GAC-OSLHF method shows a much better convergence behavior. Moreover, the GAC-OSLHF method as a Kohn-Sham method, in contrast to the ROHF approach, represents a framework which allows also for a treatment of correlation besides an exchange by appropriate functionals. In contrast to the common time-dependent density-functional methods, the GAC-OSLHF approach is capable of treating doubly or multiply excited states and can be easily applied to molecules with an open-shell ground state. On the nodal planes of the energetically highest occupied orbital, the local multiplicative GAC-OSLHF exchange potential asymptotically approaches a different, i.e., nonzero, value than in other regions, an asymptotic behavior which is known from exact Kohn-Sham exchange potentials of ground states of molecules. PMID- 16035742 TI - Electronic quantum Monte Carlo calculations of atomic forces, vibrations, and anharmonicities. AB - Atomic forces are calculated for first-row monohydrides and carbon monoxide within electronic quantum Monte Carlo (QMC). Accurate and efficient forces are achieved by using an improved method for moving variational parameters in variational QMC. Newton's method with singular value decomposition (SVD) is combined with steepest-descent (SD) updates along directions rejected by the SVD, after initial SD steps. Dissociation energies in variational and diffusion QMC agree well with the experiment. The atomic forces agree quantitatively with potential-energy surfaces, demonstrating the accuracy of this force procedure. The harmonic vibrational frequencies and anharmonicity constants, derived from the QMC energies and atomic forces, also agree well with the experimental values. PMID- 16035743 TI - Two-photon absorption in solution by means of time-dependent density-functional theory and the polarizable continuum model. AB - We present the first study of two-photon absorption (TPA) of solvated molecules based on direct evaluation of TPA cross sections from the quadratic response of time-dependent perturbations. A set of prototypical two-photon (TP) chromophores has been selected and analyzed: a pure pi system (t-stilbene) and its substituted homologs obtained employing a donor (D) and an acceptor (A) group to probe the solvent effects along the series pi, D-pi-D, A-pi-D, and A-pi-A. For the selected systems we have calculated the TPA cross sections in different environments by means of the polarizable continuum model. The data have been analyzed to evaluate how the structural and environmental parameters contribute to the final two photon absorption cross section. These include molecular structure, geometry relaxation in solution, polarity, and refractive index of the solvent. The performances of the three common functionals SVWN, BLYP, and B3LYP have been compared. The results show a significant solvent dependence of the TPA cross section and an unusual trend when passing from cyclohexane to water. The data have also been rationalized in terms of the main orbital excitations leading to the transitions. Finally, trends along the series have been described and comparison with experiments and previous calculations has been drawn. PMID- 16035744 TI - Statistical mechanical theory for steady-state systems. III. Heat flow in a Lennard-Jones fluid. AB - A statistical mechanical theory for heat flow is developed based upon the second entropy for dynamical transitions between energy moment macrostates. The thermal conductivity, as obtained from a Green-Kubo integral of a time correlation function, is derived as an approximation from these more fundamental theories, and its short-time dependence is explored. A new expression for the thermal conductivity is derived and shown to converge to its asymptotic value faster than the traditional Green-Kubo expression. An ansatz for the steady-state probability distribution for heat flow down an imposed thermal gradient is tested with simulations of a Lennard-Jones fluid. It is found to be accurate in the high density regime at not too short times, but not more generally. The probability distribution is implemented in Monte Carlo simulations, and a method for extracting the thermal conductivity is given. PMID- 16035745 TI - Novel Monte Carlo scheme for systems with short-ranged interactions. AB - We propose a Monte Carlo (MC) sampling algorithm to simulate systems of particles interacting via very short-ranged discontinuous potentials. Such models are often used to describe protein solutions or colloidal suspensions. Most normal MC algorithms fail for such systems because, at low temperatures, they tend to get trapped in local potential-energy local minima due to the short range of the pair potential. To circumvent this problem, we have devised a scheme that changes the construction of trial moves in such a way that the potential-energy difference between initial and final states drops out of the acceptance rule for the Monte Carlo trial moves. This approach allows us to simulate systems with short-ranged attraction under conditions that were unreachable up to now. PMID- 16035746 TI - Computing resonance energies, widths, and wave functions using a Lanczos method in real arithmetic. AB - We introduce new ideas for calculating resonance energies and widths. It is shown that a non-Hermitian-Lanczos approach can be used to compute eigenvalues of H+W, where H is the Hamiltonian and W is a complex absorbing potential (CAP), without evaluating complex matrix-vector products. This is done by exploiting the link between a CAP-modified Hamiltonian matrix and a real but nonsymmetric matrix U suggested by Mandelshtam and Neumaier [J. Theor. Comput. Chem. 1, 1 (2002)] and using a coupled-two-term Lanczos procedure. We use approximate resonance eigenvectors obtained from the non-Hermitian-Lanczos algorithm and a very good CAP to obtain very accurate energies and widths without solving eigenvalue problems for many values of the CAP strength parameter and searching for cusps. The method is applied to the resonances of HCO. We compare properties of the method with those of established approaches. PMID- 16035747 TI - Local correlation potentials from Brueckner coupled-cluster theory. AB - Local correlation potentials have been obtained from the nonlocal Brueckner coupled-cluster correlation potentials for the rare-gas atoms He, Ne, and Ar and the CO molecule. It is shown that the local correlation potential can mainly be expressed as a sum of two components: a "pure" correlation part and a relaxation contribution. While the total correlation potentials show an oscillating behavior near the nuclei, indicating the atomic shell structure, their components decrease rather monotonously, with a step structure in case of Ne and Ar. By looking at the determinantal overlap and one-electron properties it has been found that the orbitals obtained from these local potentials form a determinant which very well corresponds with the Brueckner determinant. Thus the previously found closeness between the Hartree-Fock determinant and the exchange-only Kohn-Sham determinant [Della Sala and Gorling, J. Chem. Phys. 115, 5718 (2001)] is confirmed also for the correlated case. PMID- 16035748 TI - Gradient-corrected density-functional potential with correct asymptotic behavior: application to interconfigurational energies in transition-metal atoms. AB - Based upon the optimized effective potential with the self-interaction correction, we present in this paper an alternative gradient-corrected density functional approximation with the proper long-range behavior of the effective potential. As applied to the study of the interconfigurational energies of the whole transition-metal atoms, the present combination of the gradient-corrected contribution and the modified optimized effective potential lead the s ionization to the excellent agreement with the experiment. The calculated d ionizations and s-d transition energies are also discussed. PMID- 16035749 TI - Density functional energy decomposition into one- and two-atom contributions. AB - The present work provides a generalization of Mayer's energy decomposition for the density-functional theory (DFT) case. It is shown that one- and two-atom Hartree-Fock energy components in Mayer's approach can be represented as an action of a one-atom potential V(A) on a one-atom density rho(A) or rho(B). To treat the exchange-correlation term in the DFT energy expression in a similar way, the exchange-correlation energy density per electron is expanded into a linear combination of basis functions. Calculations carried out for a number of density functionals demonstrate that the DFT and Hartree-Fock two-atom energies agree to a reasonable extent with each other. The two-atom energies for strong covalent bonds are within the range of typical bond dissociation energies and are therefore a convenient computational tool for assessment of individual bond strength in polyatomic molecules. For nonspecific nonbonding interactions, the two-atom energies are low. They can be either repulsive or slightly attractive, but the DFT results more frequently yield small attractive values compared to the Hartree-Fock case. The hydrogen bond in the water dimer is calculated to be between the strong covalent and nonbonding interactions on the energy scale. PMID- 16035750 TI - Coarse graining using pretabulated potentials: liquid benzene. AB - The large length and time scales involved in polymer simulation render the atomistic representation of polymer systems a computationally expensive and unnecessarily detailed procedure. We present a novel coarse-graining method for the description of nonbonded interactions between moieties composing the monomeric units of polymers, phenyl rings in particular. The method is based on the determination of the interactions between pairs of moieties from precalculated and tabulated values of the energy between the moieties in their atomistic representation. Validation of the method is performed by carrying out coarse-grained and fully atomistic simulations of a benzene liquid, where structural and thermodynamic properties at various state points are compared. The effects of the coarse grained model assumptions and of the energy table dimension and discretization are investigated. Results are also presented for the reverse mapping from the coarse grained to the fully atomistic representation. PMID- 16035751 TI - The vibrational predissociation spectra of the H5O2 +RGn(RG = Ar,Ne) clusters: correlation of the solvent perturbations in the free OH and shared proton transitions of the Zundel ion. AB - Predissociation spectra of the H(5)O(2) (+)RG(n)(RG = Ar,Ne) cluster ions are reported in energy regions corresponding to both the OH stretching (3350-3850 cm( 1)) and shared proton (850-1950 cm(-1)) vibrations. The two free OH stretching bands displayed by the Ne complex are quite close to the band origins identified earlier in bare H(5)O(2) (+) [L. I. Yeh, M. Okumura, J. D. Myers, J. M. Price, and Y. T. Lee, J. Chem. Phys. 91, 7319 (1989)], indicating that the symmetrical H(5)O(2) (+) "Zundel" ion remains largely intact in H(5)O(2) (+)Ne. The low energy spectrum of the Ne complex is simpler than that observed previously for H(5)O(2) (+)Ar, and is dominated by two sharp transitions at 928 and 1047 cm(-1), with a weaker feature at 1763 cm(-1). The H(5)O(2) (+)Ar(n),n = 1-5 spectra generally exhibit complex band structures reflecting solvent-induced symmetry breaking of the Zundel core ion. The extent of solvent perturbation is evaluated with electronic structure calculations, which predict that the rare gas atoms should attach to the spectator OH groups of H(5)O(2) (+) rather than to the shared proton. In the asymmetric complexes, the shared proton resides closer to the more heavily solvated water molecule, leading to redshifts in the rare gas atom-solvated OH stretches and to blueshifts in the shared proton vibrations. The experimental spectra are compared with recent full-dimensional vibrational calculations (diffusion Monte Carlo and multimode/vibrational configuration interaction) on H(5)O(2) (+). These results are consistent with assignment of the strong low-energy bands in the H(5)O(2) (+)Ne spectrum to the vibration of the shared proton mostly along the O-O axis, with the 1763 cm(-1) band traced primarily to the out-of-phase, intramolecular bending vibrations of the two water molecules. PMID- 16035752 TI - Two-color resonantly enhanced multiphoton ionization and zero-kinetic-energy photoelectron spectroscopy of jet-cooled indan. AB - We report studies of supersonically cooled indan using two-color resonantly enhanced multiphoton ionization and two-color zero-kinetic-energy photoelectron spectroscopy. With the aid of ab initio and density-functional calculations, vibrational modes of the first electronically excited state of the neutral species and those of the cation have been assigned, and the adiabatic ionization energy has been determined to be 68458 +/- 5 cm(-1). Similar to the ground state and the first electronically excited state of the neutral molecule, the ground state of the cation is also proven to be nonplanar, with an estimated barrier of 213 cm(-1) and a puckering angle of 15.0 degrees. These conclusions will be discussed in comparison with a previous study of an indan derivative 1,3 benzodioxole. PMID- 16035753 TI - Valence photoionization dynamics in circular dichroism of chiral free molecules: the methyl-oxirane. AB - The dynamical behavior of circular dichroism for valence photoionization processes in pure enantiomers of randomly oriented methyl-oxirane molecules has been studied by circularly polarized synchrotron radiation. Experimental results of the dichroism coefficient obtained for valence photoionization processes as a function of photon energy have been compared with theoretical values predicted by state-of-the-art ab initio density-functional theory. The circular dichroism measured at low electron kinetic energies was as large as 11%. Trends in the experimental dynamical behavior of the dichroism coefficients D(i)(omega) have been observed. Agreement between experimental and theoretical results permits unambiguous identification of the enantiomer and of the individual orbitals. PMID- 16035754 TI - Effect of rotational energy on the reaction Li + HF(upsilon = 0,j)-->LiF + H: an experimental and computational study. AB - In a crossed molecular-beam study we have measured angular and time-of-flight distributions of the product LiF from the reaction Li + HF(upsilon = 0)-->LiF + H at various collision energies ranging from 97 to 363 meV for three markedly different rotational state distributions of HF obtained at nozzle temperatures close to 315, 510, and 850 K. Particularly, for the low and intermediate collision energies we observe significant effects of the varying j-state populations on the shape of the product angular distributions. At 315 K an additional feature appears in the angular distributions which is interpreted as being due to scattering from HF dimers. The experimental data are compared with simulations of the monomer reaction based on extensive quasiclassical trajectory calculations on a new state-of-the-art ab initio potential energy surface. We find an overall good agreement between the theoretical simulations and the experimental data for the title reaction, especially at the highest HF nozzle temperature. PMID- 16035755 TI - A new ab initio potential-energy surface of HO2(X2A") and quantum studies of HO2 vibrational spectrum and rate constants for the H + O2 <--> O + OH reactions. AB - A new global potential-energy surface for the ground electronic state of HO(2)(X(2)A(")) has been developed by three-dimensional cubic spline interpolation of more than 15 000 ab initio points, which were calculated at the multireference configuration-interaction level with Davidson correction using the augmented correlation-consistent polarized valence quadruple zeta basis set. Low lying vibrational states were obtained in this new potential using the Lanczos method and assigned. The calculated vibrational frequencies are in much better agreement with the available experimental band origins than those obtained from a previous potential. In addition, rate constants for the H+O(2) <--> O + OH reactions were obtained using a wave-packet-based statistical model. Reasonably good agreement with experimental data was obtained. These results demonstrate the accuracy of the potential. PMID- 16035756 TI - HF in clusters of molecular hydrogen. I. Size evolution of quantum solvation by parahydrogen molecules. AB - We present a theoretical study of the quantum solvation of the HF molecule by a small number of parahydrogen molecules, having n = 1-13 solvent particles. The minimum-energy cluster structures determined for n = 1-12 have all of the H(2) molecules in the first solvent shell. The first solvent shell closes at n = 12 and its geometry is icosahedral, with the HF molecule at the center. The quantum mechanical ground-state properties of the clusters are calculated exactly using the diffusion Monte Carlo method. The zero-point energy of (p-H(2))(n)HF clusters is unusually large, amounting to 86% of the potential well depth for n > 7. The radial probability distribution functions (PDFs) confirm that the first solvent shell is complete for n = 12, and that the 13th p-H(2) molecule begins to fill the second solvent shell. The p-H(2) molecules execute large-amplitude motions and are highly mobile, making the solvent cage exceptionally fluxional. The anisotropy of the solvent, very pronounced for small clusters, decreases rapidly with increasing n, so that for n approximately 8-9 the solvent environment is practically isotropic. The analysis of the pair angular PDF reveals that for a given n, the parahydrogen solvent density around the HF is modulated in a pattern which clearly reflects the lowest-energy cluster configuration. The rigidity of the solvent clusters displays an interesting size dependence, increasing from n = 6 to 9, becoming floppier for n = 10, and increasing again up to n = 12, as the solvent shell is filled. The rigidity of the solvent cage appears to reach its maximum for n = 12, the point at which the first solvent shell is closed. PMID- 16035757 TI - Ab initio study of small acetonitrile cluster anions. AB - Ab initio electronic structure calculations have been performed for (CH(3)CN)(2) (-) and (CH(3)CN)(3) (-) cluster anions using a diffuse basis set. We found both the dipole-bound structures and internal structures, where in the former structure an excess electron is mainly distributed on the surface of the cluster while an excess electron is internally trapped in the latter configuration. The optimized structures found for cluster anions were compared to those for neutral clusters. Potential-energy surfaces were also plotted as a function of appropriate internal coordinates in order to understand the interconversions of the optimized structures of clusters. The relative stabilities of the optimized confirmers have been discussed on the basis of the characteristics of these potential surfaces, relative energies, and electron vertical detachment energies. PMID- 16035758 TI - The 4051-Angstroms band of C3 (A 1Piu-X 1Sigmag +, 000-000): perturbed low-J lines and lifetime measurements. AB - Rotational analyses have been carried out at high resolution for the 000-000 and 000-100 bands of the A (1)Pi(u)-X (1)Sigma(g) (+) transition of supersonic jet cooled C(3). Two different spectra have been recorded for each band, using time gatings of 20-150 and 800-2300 ns. At the shorter time delay the spectra show only the lines observed by many previous workers. At the longer time delay many extra lines appear, some of which have been observed previously by [McCall et al.Chem. Phys. Lett. 374, 583 (2003)] in cavity ring-down spectra of jet-cooled C(3). Detailed analysis of these extra lines shows that at least two long-lived states perturb the A (1)Pi(u), 000 state. One of these appears to be a (3)Sigma(u) (-) vibronic state, which may possibly be a high vibrational level of the b (3)Pi(g) state, and the other appears to be a P = 1 state with a low rotational constant B. Our spectra also confirm the reassignment by McCall et al. of the R(0) line of the 000-000 band, which is consistent with the spectra recorded towards a number of stars that indicate the presence of C(3) in the interstellar medium. Fluorescence lifetimes have been measured for a number of upper-state rotational levels. The rotational levels of the A (1)Pi(u) state have lifetimes in the range of 230-190 ns, decreasing slightly with J; the levels of the perturbing states have much longer lifetimes, with some of them showing biexponential decays. An improved value has been obtained for the nu(1) vibrational frequency of the ground state, nu(1) = 1224.4933 +/- 0.0029 cm(-1). PMID- 16035759 TI - Structure and energy difference of two isomers of He-CH3F. AB - The intermolecular potential surface of He-CH(3)F is investigated through ab initio calculations and microwave and millimeter-wave spectroscopies. The intermolecular potential is calculated at the fourth-order Moller-Plesset level with a large basis set including bond functions. Three minimums exist, the deepest of which is at the carbon end of the C-F axis and has a depth of 46.903 cm(-1), the second deepest is in a T-shaped position relative to the C-F axis with a depth of 44.790 cm(-1), and the shallowest is at the fluorine end of the C F axis with a depth of 30.929 cm(-1). The barrier to internal rotation of the CH(3)F subunit about its C-F axis is very low, thus leading to essentially free internal rotation and two separate sets of bound states correlating to ortho CH(3)F (|K| = 3n) for the ground, or A, internal rotor state upon which this study focuses, and to para-CH(3)F (|K| = 3n +/- 1) for the excited, or E, internal rotor state. Bound-state calculations of the A state performed using two different techniques show the lowest-energy state to have the helium localized in the T-shaped well with an energy of -11.460 cm(-1), while two excited configurations of the A state have the helium localized either in the well at the carbon end ("linear") with an energy of -7.468 cm(-1) or in the well at the fluorine end ("antilinear") with an energy of -4.805 cm(-1). Spectroscopic observations confirm the predicted energy-level structure of the ground and first excited states. Sixteen transitions between 12 distinct energy levels have been observed, including pure rotational transitions of both the T-shaped ground state and the linear excited state, as well as rovibrational transitions between the ground state and the linear excited state. The energy difference between the T shaped state and the linear state is measured to be 132 374.081(16) MHz. There is significant Coriolis mixing of the ground state J(K(a)K(c)) = 2(20) and the linear J(K) = 2(0) levels which aided in the observation of the T to linear transitions. This mixing and the T to linear energy difference are sensitive probes of the relative well depths of the two lowest minimums and are well predicted by the ab initio potential. Improved agreement between experiment and theory is obtained by morphing the correlation energy of the potential. He-CH(3)F is one of just a few atom-molecule complexes for which the ground-state geometry does not coincide with the global potential minimum. PMID- 16035760 TI - Decomposition of nitramine energetic materials in excited electronic states: RDX and HMX. AB - Ultraviolet excitation (8-ns duration) is employed to study the decomposition of RDX (1,3,5-trinitro-1,3,5-triazacyclohexane) and HMX (1,3,5,7-tetranitro-1,3,5,7 tetrazacyclooctane) from their first excited electronic states. Isolated RDX and HMX are generated in the gas phase utilizing a combination of matrix-assisted laser desorption and supersonic jet expansion techniques. The NO molecule is observed as one of the initial dissociation products by both time-of-flight mass spectroscopy and laser-induced fluorescence spectroscopy. Four different vibronic transitions of NO are observed: A (2)Sigma(v(') = 0)<--X (2)Pi(v(") = 0,1,2,3). Simulations of the NO rovibronic intensities for the A<--X transitions show that dissociated NO from RDX and HMX is rotationally cold (approximately 20 K) and vibrationally hot (approximately 1800 K). Another potential initial product of RDX and HMX excited state dissociation could be OH, generated along with NO, perhaps from a HONO intermediate species. The OH radical is not observed in fluorescence even though its transition intensity is calculated to be 1.5 times that found for NO per radical generated. The HONO intermediate is thereby found not to be an important pathway for the excited electronic state decomposition of these cyclic nitramines. PMID- 16035761 TI - Absolute cross sections of ozone at atmospheric temperatures for the Wulf and the Chappuis bands. AB - Ozone absorption constitutes a variable background against which measurement of pollution is observed from satellites by the solar occultation technique. The temperature-altitude gradient of ozone spans the range 180-340 K. Laboratory measurements of ozone absorption at temperatures across this range are needed to calibrate spectroscopic remote sensing, but have yielded results in substantial disagreement with each other. This paper presents the first measurements of the variation of absorption cross-sections of ozone at temperatures from 144 to 300 K over the 9250-18 500 cm(-1) spectral region. PMID- 16035762 TI - High-resolution ultraviolet spectroscopy of p-fluorostyrene-water: evidence for a sigma-type hydrogen-bonded dimer. AB - Ab initio calculations predict four stable conformational structures of the singly hydrated cluster of p-fluorostyrene: two out of plane with pi- and two in plane with sigma-type intermolecular hydrogen bonding between p-fluorostyrene and water. We employed mass-selective resonance-enhanced two-photon ionization high resolution (70-MHz FWHM laser bandwidth) spectroscopy to partially resolve the rotational structure of the 0(0) (0) origin band of the S(1) <--S(0) electronic transition. A computer-aided fit based on genetic algorithms was used to analyze the experimental high-resolution spectrum and to determine the observed conformational structure. The good agreement between the experimental and the simulated spectra of the 0(0) (0) band and the assignment of the other prominent bands as inter- and intramolecular vibrational progressions clearly demonstrates that the anti in-plane conformer is the most abundant one in the molecular beam. The existence of the sigma-type hydrogen bond between p-fluorostyrene and water manifests that the electron attracting effect of fluorine dominates over the releasing mesomeric effect of the vinyl group and thus a pi-type hydrogen bonding with the aromatic ring is not favored in this case. PMID- 16035763 TI - Fluorescence-dip infrared spectroscopy and predissociation dynamics of OH A 2Sigma+ (v = 4) radicals. AB - Fluorescence-dip infrared spectroscopy, an UV-IR double-resonance technique, is employed to characterize the line positions, linewidths, and corresponding lifetimes of highly predissociative rovibrational levels of the excited A (2)Sigma(+) electronic state of the OH radical. Various lines of the 4 <--2 overtone transition in the excited A (2)Sigma(+) state are observed, from which the rotational, centrifugal distortion, and spin-rotation constants for the A (2)Sigma(+) (v = 4) state are determined, along with the vibrational frequency for the overtone transition. Homogeneous linewidths of 0.23-0.31 cm(-1) full width at half maximum are extracted from the line profiles, demonstrating that the N = 0 to 7 rotational levels of the OH A (2)Sigma(+) (v = 4) state undergo rapid predissociation with lifetimes of < or =23 ps. The experimental linewidths are in near quantitative agreement with first-principles theoretical predictions. PMID- 16035764 TI - Experimental and theoretical studies of rate coefficients for the reaction O(3P)+CH3OH at high temperatures. AB - Rate coefficients of the reaction O((3)P) + CH(3)OH in the temperature range of 835-1777 K were determined using a diaphragmless shock tube. O atoms were generated by photolysis of SO(2) with a KrF excimer laser at 248 nm or an ArF excimer laser at 193 nm; their concentrations were monitored via atomic resonance absorption excited by emission from a microwave-discharged mixture of O(2) and He. The rate coefficients determined for the temperature range can be represented by the Arrhenius equation, k(T) = (2.29 +/- 0.18) x 10(-10) exp[-(4210 +/- 100)T] cm(3) molecule(-1) s(-1); unless otherwise noted, all the listed errors represent one standard deviation in fitting. Combination of these and previous data at lower temperature shows a non-Arrhenius behavior described as the three-parameter equation, k(T) = (2.74 +/- 0.07) x 10(-18)T(2.25 +/- 0.13) exp[-(1500 +/- 90)T] cm(3)molecule(-1) s(-1). Theoretical calculations at the Becke-3-Lee-Yang-Parr (B3LYP)6-311 + G(3df,2p) level locate three transition states. Based on the energies computed with coupled clusters singles, doubles (triples) [CCSD(T)]/6 311 + G(3df,2p)B3LYP6-311 + G(3df,2p), the rate coefficients predicted with canonical variational transition state theory with small curvature tunneling corrections agree satisfactorily with the experimental observations. The branching ratios of two accessible reaction channels forming OH + CH(2)OH (1a) and OH + CH(3)O (1b) are predicted to vary strongly with temperature. At 300 K, reaction (1a) dominates, whereas reaction (1b) becomes more important than reaction (1a) above 1700 K. PMID- 16035765 TI - Ultrafast proton transfer of 1-hydroxy-2-acetonaphthone: reaction path from resonance Raman and transient absorption studies. AB - The intramolecular degrees of freedom contributing to the ultrafast excited-state intramolecular proton transfer of 1-hydroxy-2-acetonaphthone are determined. Thereto, resonance Raman studies are combined with transient absorption measurements with 30-fs time resolution. Enhanced Raman intensity is found in coordinates that are dominantly associated with deformations and bond length changes in the naphthalene chromophore. This indicates that the primary changes after the optical excitation are a geometric relaxation of the chromophore. A ringing of the molecule after the ultrafast proton transfer is observed by the transient absorption measurements. It reveals the nuclear coordinates contributing to the reaction path beyond the Franck-Condon region. There, planar H-chelate ring deformations changing the donor-acceptor distance are found to dominate. The difference in the observed vibrational signatures indicates a significant turn in the reaction path. PMID- 16035766 TI - Exploratory study of the electron-density relocalization process in benzene through a time-dependent topological analysis. AB - A time-dependent topological analysis of the electron density in benzene provides quantitative information about charge oscillation processes, and particularly about the two highest pi orbitals contributions to carbon topological basins. In this exploratory study coupling one molecular-dynamic trajectory to a topological analysis, Fourier transform of the topological time data autocorrelation functions indicates frequencies of oscillations, and shows that carbon atoms, though identical in average, would be more or less highest occupied molecular orbital reactive with a cyclic behavior. PMID- 16035767 TI - Spectroscopic investigation of the solvation of organic molecules in superfluid helium droplets. AB - The spectroscopy of molecules doped into superfluid helium nanodroplets provides valuable information on the process of solvation in superfluid helium. In continuation of an earlier report on emission spectra of various phthalocyanines showing a splitting of all molecular transitions in the range of about 5-12 cm( 1), the emission spectra of tetracene, pentacene, and perylene in superfluid helium droplets are presented. The new spectra and the results obtained for the phthalocyanines are explained by an empirical model which accounts for the existence of different metastable configurations of a nonsuperfluid solvation layer around the guest molecule. PMID- 16035768 TI - Evidence for new bands in the 3nu1 and 4nu1 regions of propyne. AB - Vibrationally mediated photodissociation and room-temperature photoacoustic (PA) spectroscopy have been used for obtaining action (monitoring the yield of H photofragments) and absorption spectra of the second (3nu(1)) and third (4nu(1)) C-H acetylenic stretches overtone regions in propyne. The band contours appearing in these regions seem mostly regular even though they are perturbed, as expressed by the origin shifts in different K components, splitting of the K structure, and splitting due to resonances between neighboring states. Symmetric rotor simulations of the band contours of the PA and action spectra allowed extraction of the molecular parameters and rough estimates for the homogeneous broadening arising from energy flow to the bath vibrational states. We particularly benefited from the reduced congestion in the jet-cooled action spectra and their simulations, which enabled observation of yet unknown features in the vicinity of the 3nu(1) and 4nu(1) states. Particularly, the emergence of the new state in the 3nu(1) region was confirmed by the action spectra monitored at several differing jet temperatures, suggesting that it is a dark state in IR vibrational excitation that becomes brighter in UV excitation to the upper electronic state. The monitored and Gaussian-fitted Doppler profiles point to the release of H photofragments with low average translational energies, attributed to an indirect dissociation process occurring after internal conversion to the ground electronic state and isomerization to allene. PMID- 16035769 TI - Vibrational assignment and Franck-Condon analysis of the mass-analyzed threshold ionization (MATI) spectrum of CH2ClI: the effect of strong spin-orbit interaction. AB - Detailed analysis of the one-photon mass-analyzed threshold ionization (MATI) spectrum of CH(2)ClI is presented. This includes the determination of the ionization energy of CH(2)ClI, complete vibrational assignments, and quantum chemical calculations at the spin-orbit density-functional-theory (SODFT) level with various basis sets. Relativistic effective core potentials with effective spin-orbit operators can be used in SODFT calculations to treat the spin-orbit term on an equal footing with other relativistic effects and electron correlations. The comparison of calculated and experimental vibrational frequencies indicate that the spin-orbit effects are essential for the reasonable description of the CH(2)ClI(+) cation. Geometrical parameters and thus the molecular shape of the cation are greatly influenced by the spin-orbit effects even for the ground state. Calculated geometrical parameters deviate substantially for different basis sets or effective core potentials. In an effort to derive the exact geometrical parameters for this cation, SODFT geometries were further improved utilizing Franck-Condon fit of the MATI spectral pattern. This empirical fitting produced the well-converged set of geometrical parameters that are quite insensitive to the choice of SODFT calculations. The C-I bond length and the Cl-C-I bond angle show large deviations among different SODFT calculations, but the empirical spectral fitting yields 2.191 +/- 0.003 Angstroms for the C-I bond length and 107.09 +/- 0.09 degrees for the Cl-C-I angle. Those fitted geometrical parameters along with the experimental vibrational frequencies could serve as a useful reference in calibrating relativistic quantum-chemical methods for radicals. PMID- 16035770 TI - Study of 7-azaindole in its first four singlet states. AB - The molecular structure and properties of 7-azaindole in its first four singlet states were studied with a view to improving current understanding of the photophysical behavior of its C(2h) dimer. This dimer, which exhibits a double proton transfer via its two hydrogen bonds upon electronic excitation, has for 35 years been used as a model for the photophysical behavior of DNA base pairs. Electronic excitation of 7-azaindole simultaneously increases its acidity and basicity; these changes facilitate a concerted mechanism for the double proton transfer in the dimer. In this work, we found the acidity and basicity changes to occur only in its first pi,pi(*) excited singlet state. PMID- 16035771 TI - Photolysis wavelength dependence of the translational anisotropy and the angular momentum polarization of O2(a 1Deltag) formed from the UV photodissociation of O3. AB - The translational anisotropy and angular momentum polarization of the O(2)(a (1)Delta(g),v = 0;J = 15-27) molecular photofragment produced from the UV photodissociation of O(3) in the range from 270 to 300 nm have been determined using resonance-enhanced multiphoton ionization in conjunction with time-of flight mass spectrometry. At the shortest photolysis wavelengths used, the fragments exhibit the anisotropic vector correlations expected from a prompt dissociation via the (1)B(2) <--(1)A(1) transition. Deviations from this behavior are observed at longer photolysis wavelengths with, in particular, the angular momentum orientation showing a significant reduction in magnitude. This indicates that the dissociation can no longer be described by a purely impulsive model and a change in geometry of the dissociating molecule is implied. This observation is substantiated by the variation of the translational anisotropy with photolysis wavelength. We also observe that the bipolar moments describing the angular momentum polarization of the odd J states probed are consistently lower in magnitude than those of the even J states and that this variation is observed for all photolysis wavelengths. PMID- 16035772 TI - A time-dependent wave-packet quantum scattering study of the reaction H2+(v = 0 2,4,6;j = 1) + He--> HeH+ + H. AB - The quantum scattering dynamics calculation was carried out for the titled reaction in the collision energy range of 0.0-2.4 eV with reactant H(2) (+) in the rotational state j = 1 and vibrational states v = 0-2, 4, and 6. The present time-dependent wave-packet calculation takes into account the Coriolis coupling (CC) and uses the accurate ab initio potential-energy surface of Palmieri et al. [Mol. Phys. 98, 1835 (2000)]. The importance of including the CC quantum scattering calculation has been revealed by the comparison between the CC calculation and the previous coupled state (CS) calculation. The CC total cross sections for the v = 2, 4, and 6 states show collision energy-dependent behaviors different from those based on the CS calculation. Furthermore, the collision energy dependence of the total cross sections obtained in the present CC calculation only exhibits minor oscillations, indicating that the chance is slim for reactive resonances in total cross sections to survive through the partial wave averaging. The magnitude and profile of the CC total cross sections for v = 0-2 in the collision energy range of 0.0-2.5 eV are found to be consistent with experimental cross sections obtained recently by Tang et al. [J. Chem. Phys. 122, 164301 (2005)] after taking into account the experimental uncertainties. PMID- 16035773 TI - Collision-induced conformational changes in glycine. AB - We present quantum dynamical calculations on the conformational changes of glycine in collisions with the He, Ne, and Ar rare-gas atoms. For two conformer interconversion processes (III-->I and IV-->I), we find that the probability of interconversion is dependent on several factors, including the energy of the collision, the angle at which the colliding atom approaches the glycine molecule, and the strength of the glycine-atom interaction. Furthermore, we show that attractive interactions between the colliding atom and the glycine molecule catalyze conformer interconversion at low collision energies. In previous infrared spectroscopy studies of glycine trapped in rare-gas matrices and helium clusters, conformer III has been consistently observed, but conformer IV has yet to be conclusively detected. Because of the calculated thermodynamic stability of conformer IV, its elusiveness has been attributed to the IV-->I conformer interconversion process. However, our calculations present little indication that IV-->I interconversion occurs more readily than III-->I interconversion. Although we cannot determine whether conformer IV interconverts during experimental Ne- and Ar-matrix depositions, our evidence suggests that the conformer should be present in helium droplets. Anharmonic vibrational frequency calculations illustrate that previous efforts to detect conformer IV may have been hindered by the overlap of its IR-absorption bands with those of other conformers. We propose that the redshifted symmetric -CH(2) stretch of conformer IV provides a means for its conclusive experimental detection. PMID- 16035774 TI - Charge-induced spin alignment in diradical donor molecules: numerical calculations of correlated many-electron-spin systems. AB - The mechanism of charge-induced high spin is studied in pi-conjugated molecules by means of a model-Hamiltonian approach. Intersite Coulomb interactions are taken into account in a pi-conjugated moiety, which is coupled with two localized spins through exchange interactions. We clarify spin alignment in neutral and oxidized states by exact numerical calculations including all the correlation effects. In thianthrene-based molecules, one-electron oxidation induces strong ferromagnetic correlation between the localized spins irrespective of the spin alignment in the neutral state. The localized spins are coupled to the delocalized hole spin ferromagnetically, leading to a high-spin state in the oxidized molecule. Our calculations on structural dependence and effective exchange interaction are consistent with the recent experiment of thianthrene bis(nitronyl nitroxide). By comparing the thianthrene-based molecule with the anthracene-based one, we clarify the role of superexchange interactions via the sulfur atoms. PMID- 16035775 TI - Ab initio computed diabatic potential energy surfaces of OH-HCl. AB - The two four-dimensional diabatic potential energy surfaces (DPESs) for OH-HCl are computed that correlate with the twofold degenerate (2)Pi ground state of the free OH radical. About 20 000 points on the surface are obtained by the ab initio coupled-cluster and multi-reference configuration interaction methods. Analytic forms for the diabatic potential energy surfaces are derived as expansions in complete sets of orthogonal functions depending on the three intermolecular angles. The numeric computation of the angular expansion coefficients is discussed. The distance-dependence of the angular coefficients is represented by the reproducing kernel Hilbert space method. It is checked that both diabatic potentials converge for large intermolecular separations to the values computed directly from the electrostatic multipole expansion. The final DPESs are discussed and illustrated by some physically meaningful one- and two-dimensional cuts through them. PMID- 16035776 TI - Infrared spectra of water clusters in krypton and xenon matrices. AB - The infrared absorption spectra of the water molecules and small water clusters, (H(2)O)(n) with n = 2-6, trapped in solid argon, krypton, and xenon matrices have been investigated. The infrared bands of the water clusters with n = 5 and 6 in krypton and n = 3, 4, 5, and 6 in xenon matrices have been identified for the first time in the bonded OH stretching region. The frequency shifts in the bonded OH stretching band of the water dimer and trimer in xenon matrices show fairly large deviations to the red from the empirical correlation between the matrix shifts and the square root of the critical temperatures of the matrix material. The observed anomalous shifts suggest that the water dimer and trimer in solid xenon are trapped in multiple sites, and that the structures of the preferential trapping sites are different from those in argon and krypton matrices. PMID- 16035777 TI - Qualitative features of the two-dimensional Raman spectrum in liquids. AB - The theory presented earlier [J. Kim and T. Keyes, Phys. Rev. E 66, 051110 (2002)] is analyzed to determine the information available from the two dimensional Raman spectrum R((5))(t(2),t(1)) in liquids. The known spectra are well represented by the sum of two products of ordinary time correlations predicted by the theory. The shape of R((5)) is related in general to the values of simple same-time averages and concepts amenable to physical intuition. Using standard models for the time correlations entering the theory, specific analytic expressions for the spectrum are obtained depending on two parameters and a time scale, and the behavior of the spectrum is mapped out in the parameter space. PMID- 16035778 TI - A new ultrafast technique for measuring the terahertz dynamics of chiral molecules: the theory of optical heterodyne-detected Raman-induced Kerr optical activity. AB - Optical heterodyne-detected Raman-induced Kerr optical activity (OHD-RIKOA) is a nonresonant ultrafast chiroptical technique for measuring the terahertz-frequency Raman spectrum of chirally active modes in liquids, solutions, and glasses of chiral molecules. OHD-RIKOA has the potential to provide much more information on the structure of molecules and the symmetries of librational and vibrational modes than the well-known nonchirally sensitive technique optical heterodyne detected Raman-induced Kerr-effect spectroscopy (OHD-RIKES). The theory of OHD RIKOA is presented and possible practical ways of performing the experiments are analyzed. PMID- 16035779 TI - Properties of the H-bond network for two-dimensional lattice water model. AB - A microscopic Hamiltonian of the hydrogen-bond network in two-dimensional lattice water is proposed, which describes the formation and disruption of the H bonds, their bending, and which satisfies the Bernal-Fowler rules [J. D. Bernal and R. H. Fowler, J. Chem. Phys. 1, 515 (1933)]. The thermodynamic properties of the H bond network are studied using the method of many-particle irreducible distribution functions, which is a generalization of the Kikuchi cluster approach [R. Kikuchi, Phys. Rev. 81, 988 (1951)] and the Bethe-Peierls quasiactivities method [H. A. Bethe, Prog. R. Soc. A 150, 552 (1935)]. The temperature dependencies of the average number of H bonds per molecules, the contribution of the H bonds into the heat capacity of the system, and the parameters describing the correlations between the states of molecules on the neighboring sites are investigated. It is shown that depending on the magnitude of the interaction between the H bonds in the H-bond subsystem either smooth or sharp first-order phase transition can occur. The role of different factors in the formation of the properties of the H-bond network is discussed. PMID- 16035780 TI - Quantum-classical Liouville dynamics of nonadiabatic proton transfer. AB - A proton transfer reaction in a linear hydrogen-bonded complex dissolved in a polar solvent is studied using mixed quantum-classical Liouville dynamics. In this system, the proton is treated quantum mechanically and the remainder of the degrees of freedom is treated classically. The rates and mechanisms of the reaction are investigated using both adiabatic and nonadiabatic molecular dynamics. We use a nonadiabatic dynamics algorithm which allows the system to evolve on single adiabatic surfaces and on coherently coupled pairs of adiabatic surfaces. Reactive-flux correlation function expressions are used to compute the rate coefficients and the role of the dynamics on the coherently coupled surfaces is elucidated. PMID- 16035781 TI - Dynamics of fluids near the consolute critical point: a molecular-dynamics study of the Widom-Rowlinson mixture. AB - Molecular-dynamics simulations are presented for the dynamic behavior of the Widom-Rowlinson mixture [B. Widom, and J. S. Rowlinson, J. Chem. Phys. 52, 1670 (1970)] at its critical point. This model consists of two components where like species do not interact and unlike species interact via a hard-core potential. Critical exponents are obtained from a finite-size scaling analysis. The self diffusion coefficient shows no anomalous behavior near the critical point. The shear viscosity and thermal conductivity show no divergent behavior for the system sizes considered, although there is a significant critical enhancement. The mutual diffusion coefficient, D(AB), vanishes as D(AB) approximately xi(-1.26 +/- 0.08), where xi is the correlation length. This is different from the renormalization-group (D(AB) approximately xi(-1.065)) mode coupling theory (D(AB) approximately xi(-1)) predictions. The theories and simulations can be reconciled if we assume that logarithmic corrections to scaling are important. PMID- 16035782 TI - Hydration structure of Y3+ and La3+ compared: an application of metadynamics. AB - We studied the hydration structures of Y(3+) and La(3+) in aqueous solutions by applying the metadynamics method recently introduced as a tool to explore reaction pathways based on the Car-Parrinello molecular dynamics. By employing the metal-water oxygen coordination number as a collective variable of the metadynamics a couple of aqua and chloroaqua complexes are successfully generated within the time scale of 10 ps. The reconstructed free-energy surface captures the characteristics of the hydration of the light and heavy trivalent rare-earth ions. The present study demonstrates that the metadynamics based on the Car Parrinello molecular dynamics is a promising tool for exploring the free-energy surface of complicated systems such as solutions. PMID- 16035783 TI - Stokes-Einstein relations for a square-well fluid. AB - A Stokes-Einstein relation, relating the shear viscosity eta to the self diffusion coefficient D, is constructed for a classical fluid subject to an effective two-body intermolecular force, derived from a square-well potential, undergoing dynamics as described by a Smoluchowski equation for pair diffusion. The time correlation functions for eta and 1D are separated into contributions from delta function, hard-sphere forces, and from delta function, square-well soft forces. Furthermore, D is separated into its two- and three-body time correlation functions, and eta into its two- to four-body terms. D shows activated diffusion, as in Arrhenius behavior, and on the level of two-body dynamics, the Deta product adheres to the Stokes-Einstein relation, subject to a small correction for potential softness. Three-body time correlation functions increase D, whereas three- and four-body correlation functions in eta are partially offsetting. The deviation of Deta product from the Stokes-Einstein law arises from the three-body time correlations functions in D. PMID- 16035784 TI - Hydrogen bonding and dynamics of methanol by high-pressure diamond-anvil cell NMR. AB - Liquid methanol at densities up to rhorho(0) = 1.7 was studied by NMR in a specially designed diamond-anvil cell. Methyl and hydroxyl resonances have been separately observed at pressures to 43 kbars which exceeds equilibrium freezing pressure of methanol. The chemical shift difference between methyl and hydroxyl protons increases nonlinearly with increasing density, indicating a noticeable decrease in hydrogen bond length. The analyses of spin-lattice relaxation rates of both hydroxyl and methyl protons indicate that compression enhances intermolecular proton exchange and selectively reduces motion of the hydroxyl protons. Collectively these observations reveal that hydrogen bonding interaction in liquid methanol noticeably increases with compression, inhibiting the liquid solid transition even above the freezing pressure. PMID- 16035785 TI - Spherical tensor analysis of nuclear magnetic resonance signals. AB - In a nuclear magnetic-resonance (NMR) experiment, the spin density operator may be regarded as a superposition of irreducible spherical tensor operators. Each of these spin operators evolves during the NMR experiment and may give rise to an NMR signal at a later time. The NMR signal at the end of a pulse sequence may, therefore, be regarded as a superposition of spherical components, each derived from a different spherical tensor operator. We describe an experimental method, called spherical tensor analysis (STA), which allows the complete resolution of the NMR signal into its individual spherical components. The method is demonstrated on a powder of a (13)C-labeled amino acid, exposed to a pulse sequence generating a double-quantum effective Hamiltonian. The propagation of spin order through the space of spherical tensor operators is revealed by the STA procedure, both in static and rotating solids. Possible applications of STA to the NMR of liquids, liquid crystals, and solids are discussed. PMID- 16035786 TI - From dimer to condensed phases at extreme conditions: accurate predictions of the properties of water by a Gaussian charge polarizable model. AB - Water exhibits many unusual properties that are essential for the existence of life. Water completely changes its character from ambient to supercritical conditions in a way that makes it possible to sustain life at extreme conditions, leading to conjectures that life may have originated in deep-sea vents. Molecular simulation can be very useful in exploring biological and chemical systems, particularly at extreme conditions for which experiments are either difficult or impossible; however this scenario entails an accurate molecular model for water applicable over a wide range of state conditions. Here, we present a Gaussian charge polarizable model (GCPM) based on the model developed earlier by Chialvo and Cummings [Fluid Phase Equilib. 150, 73 (1998)] which is, to our knowledge, the first that satisfies the water monomer and dimer properties, and simultaneously yields very accurate predictions of dielectric, structural, vapor liquid equilibria, and transport properties, over the entire fluid range. This model would be appropriate for simulating biological and chemical systems at both ambient and extreme conditions. The particularity of the GCPM model is the use of Gaussian distributions instead of points to represent the partial charges on the water molecules. These charge distributions combined with a dipole polarizability and a Buckingham exp-6 potential are found to play a crucial role for the successful and simultaneous predictions of a variety of water properties. This work not only aims at presenting an accurate model for water, but also at proposing strategies to develop classical accurate models for the predictions of structural, dynamic, and thermodynamic properties. PMID- 16035787 TI - Wave model for conservative bound systems. AB - In the hidden variable theory, Bohm proved a connection between the Schrodinger and Hamilton-Jacobi equations and showed the existence of classical paths, for which the generalized Bohr quantization condition is valid. In this paper we prove similar properties, starting from the equivalence between the Schrodinger and wave equations in the case of the conservative bound systems. Our approach is based on the equations and postulates of quantum mechanics without using any additional postulate. Like in the hidden variable theory, the above properties are proven without using the approximation of geometrical optics or the semiclassical approximation. Since the classical paths have only a mathematical significance in our analysis, our approach is consistent with the postulates of quantum mechanics. PMID- 16035788 TI - Dynamics of propylene glycol and its oligomers confined to a single molecular layer. AB - The dynamics of propylene glycol (PG) and its oligomers 7-PG and poly-propylene glycol (PPG), with M(w) = 4000 (approximately 70 monomers), confined in a Na vermiculite clay have been investigated by quasielastic neutron scattering. The liquids are confined to single molecular layers between clay platelets, giving a true two-dimensional liquid. Data from three different spectrometers of different resolutions were Fourier transformed to S(Q,t) and combined to give an extended dynamical time range of 0.3-2000 ps. An attempt was made to distinguish the diffusive motion from the methyl group rotation and a fast local motion of hydrogen in the polymer backbone. The results show that the average relaxation time tau(d) of this diffusive process is, as expected, larger than the relaxation time tau averaged over all dynamical processes observed in the experimental time window. More interesting, it is evident that the severe confinement has a relatively small effect on tau(d) at T = 300 K, this holds particularly for the longest oligomer, PPG. The most significant difference is that the chain-length dependence of tau(d) is weaker for the confined liquids, although the slowing down in bulk PG due to the formation of a three-dimensional network of OH-bonded end groups reduces this difference. The estimated average relaxation time tau at Q = 0.92 Angstroms(-1) for all the observed processes is in excellent agreement with the previously reported dielectric alpha relaxation time in the studied temperature range of 260-380 K. The average relaxation time tau (as well as the dielectric alpha relaxation time) is also almost unaffected by the confinement to a single molecular layer, suggesting that the interaction with the clay surfaces is weak and that the reduced dimensionality has only a weak influence on the time scale of all the dynamical processes observed in this study. PMID- 16035789 TI - Conformations and charge transport characteristics of biphenyldithiol self assembled-monolayer molecular electronic devices: a multiscale computational study. AB - We report a computational study of conformations and charge transport characteristics of biphenyldithiol (BPDT) monolayers in the (sqrt.3 x sqrt.3)R30 degrees packing ratio sandwiched between Au(111) electrodes. From force-field molecular-dynamics and annealing simulations of BPDT self-assembled monolayers (SAMs) with up to 100 molecules on a Au(111) substrate, we identify an energetically favorable herringbone-type SAM packing configuration and a less stable parallel packing configuration. Both SAMs are described by the (2sqrt.3 x sqrt.3)R30 degrees unit cell including two molecules. With subsequent density functional theory calculations of one unit cell of the (i) herringbone SAM with the molecular tilt angle theta approximately 15 degrees , (ii) herringbone SAM with theta approximately 30 degrees , and (iii) parallel SAM with theta approximately 30 degrees, we confirm that the herringbone packing configuration is more stable than the parallel one but find that the energy variation with respect to the molecule tilting within the herringbone packing is very small. Next, by capping these SAMs with the top Au(111) electrode, we prepare three molecular electronic device models and calculate their coherent charge transport properties within the matrix Green's function approach. Current-voltage (I-V) curves are then obtained via the Landauer-Buttiker formula. We find that at low bias voltages (|V| < or = 0.2 V) the I-V characteristics of models (ii) and (iii) are similar and the current in model (i) is smaller than that in (ii) and (iii). On the other hand, at higher-bias voltages (|V| > or 0.5 V), the I-V characteristics of the three models show noticeable differences due to different phenyl band structures. We thus conclude that the BPDT SAM I-V characteristics in the low-bias voltage region are mainly determined by the -Au [corrected] interaction within the individual molecule-electrode contact, while both intramolecular conformation and intermolecular interaction can affect the BPDT SAM I-V characteristics in the high-bias voltage region. PMID- 16035790 TI - Orbital specific chemistry: controlling the pathway in single-molecule dissociation. AB - A scanning tunneling microscope (STM) was used to control the pathway of the dissociation of single O(2) molecules chemisorbed on Ag(110) at 13 K. Tunneling of electrons from the STM tip into the O(2) caused dissociation of the molecule, giving rise to two adsorbed O atoms separated along the [110] direction. In contrast, the ejection of electrons from the O(2) molecule produced adsorbed O atoms separated along the [001] direction. These results illustrate that control of the dissociation pathway and product formation are associated with a specific molecular orbital located at the Fermi level. PMID- 16035791 TI - Dispersion of the local parameters of quasilocalized low-frequency vibrational modes in a low-temperature glass: direct observation via single-molecule spectroscopy. AB - Spectra of single tetra-tert-butylterrylene chromophore molecules embedded in an amorphous polyisobutylene matrix as microprobes were recorded. The individual temperature dependences of the spectral linewidths for the same single molecules (SMs) in a broad temperature interval (1.6 < T < 40 K) have been measured. This enabled us to separate the contributions of tunneling two-level systems and quasilocalized low-frequency vibrational modes (LFMs) to the observed linewidths. The analysis of the T dependences yields the values of LFM frequencies and SM-LFM coupling constants for the LFMs in the local environment of a given chromophore. Pronounced distributions of the observed parameters of LFMs were found. This result can be regarded as the first direct experimental proof of the localized nature of LFMs in glasses. PMID- 16035792 TI - Kinetics of two-step nucleation of crystals. AB - When the nucleation of a stable crystalline phase directly in a supersaturated old phase is greatly retarded, the crystal nuclei might nucleate within faster forming particles of an intermediate phase. Here we present a theoretical investigation of the kinetics of this two-step nucleation of crystals and derive general expressions for the time dependence of the number of crystals nucleated within the particles of the intermediate phase. The results reveal that crystal nucleation can be strongly delayed by the slow growth of the particles and/or by the slow nucleation of the crystals in them. Furthermore, the linear part of the time dependence of the number of nucleated crystals is determined by the formation rate of the intermediate particles. This is in contrast with the one step nucleation of crystals when this linear part is determined by the rate of crystal nucleation directly in the old phase. Criteria are proposed for distinction between the one- and two-step nucleation mechanisms, based on the supersaturation dependence of the delay time for nucleation. The application of the theoretical approach to the analysis of experimental data on the nucleation of crystals and other ordered aggregates of protein and other soluble materials is discussed. PMID- 16035793 TI - Asynchronous multicanonical basin hopping method and its application to cobalt nanoclusters. AB - The multicanonical basin hopping (MUBH) method, which uses a multicanonical weight in the basin hopping (BH) Monte Carlo method, was found to be very efficient for global optimization of large-scale systems such as Lennard-Jones clusters containing more than 150 atoms. We have implemented an asynchronous parallel version of the MUBH method using the message passing interface (MPI) to take advantage of the full usage of multiprocessors in either a homogeneous or heterogeneous computational environment. Based on the intrinsic properties of the Monte Carlo method, this MPI implementation used the task parallelism to minimize interthread data communication. For a Co nanocluster consisting of N atoms, we have applied the asynchronous multicanonical basin hopping (AMUBH) method (for 181 < N < or = 200), together with BH (for 2 < or = N < 150) and MUBH (for 150 < or = N < or = 180), to search for the molecular configuration of the global energy minimum. AMUBH becomes the only practical computational scheme for locating the energy minimum within realistic computational time for a relatively large cluster. PMID- 16035794 TI - Direct computer simulation of water-mediated force between supported phospholipid membranes. AB - The grand canonical Monte Carlo technique is used to calculate the water-mediated force operating between two supported 1,2-dilauroyl-DL-phosphatidylethanolamine (DLPE) membranes in the short separation range. The intra- and intermolecular interactions in the system are described with a combination of an AMBER-based force field for DLPE and a TIP4P model for water. The long range contributions to the electrostatic interaction energy are treated in the dipole-dipole group-based approximation. The total water-mediated force is analyzed in terms of its hydration component and the component due to the direct interaction between the membranes. The latter is, in addition, partitioned into the electrostatic, van der Waals, and steric repulsion contributions to give an idea of their relative significance in the water-mediated interaction of the membranes. PMID- 16035795 TI - Effects of compositional defects on small polaron hopping in micas. AB - Hartree-Fock calculations and electron transfer (ET) theory were used to model the effects of compositional defects on ET in the brucite-like octahedral sheet of mica. ET was modeled as an Fe(IIIII) valence interchange reaction across shared octahedral edges of the M2-M2 iron sublattice. The model entails the hopping of localized electrons and small polaron behavior. Hartree-Fock calculations indicate that substitution of F for structural OH bridges increases the reorganization energy lambda, decreases the electronic coupling matrix element V(AB), and thereby substantially decreases the hopping rate. The lambda increase arises from modification of the metal-ligand bond force constants, and the V(AB) decrease arises from reduction of superexchange interaction through anion bridges. Deprotonation of an OH bridge, consistent with a possible mechanism of maintaining charge neutrality during net oxidation, yields a net increase in the ET rate. Although substitution of Al or Mg for Fe in M1 sites distorts the structure of adjacent Fe-occupied M2 sites, the distortion has little net impact on ET rates through these M2 sites. Hence the main effect of Al or Mg substitution for Fe, should it occur in the M2 sublattice, is to block ET pathways. Collectively, these findings pave the way for larger-scale oxidation/reduction models to be constructed for realistic, compositionally diverse micas. PMID- 16035796 TI - Ultrafast selected energy x-ray absorption spectroscopy investigations of Ni and Zn species. AB - The results of ultrafast selected energy x-ray absorption spectroscopy (USEXAS) investigations of Ni and Zn species are presented. The USEXAS measurements described here employed characteristic x-ray radiation of L(alpha) and L(beta) from an ultrafast laser-driven W x-ray target to probe the K absorption edges of Ni and Zn, respectively. Static x-ray absorption edge spectra of six Ni and Zn species in either solid or solution form were obtained. Simulations of near-edge x-ray absorption spectra of these Ni and Zn species were carried out with FEFF. The results of USEXAS measurements were in general agreement with the theoretically simulated spectra and those measured with synchrotron x-ray radiation. PMID- 16035797 TI - Wetting kinetics of films containing nonadsorbing polymers. AB - Kinetics of wetting by a polymer solution have been studied theoretically for a film pinned to a slot. The fluid mechanical equations have been solved using a numerical scheme. The role of polymers appears in the disjoining pressure in the model. The spreading kinetics are observed to follow a power law: a power of 14 is observed at short times due to the Laplace pressure, and 12 at large times under the Hamaker part of the disjoining pressure at very large times and with no equilibration. It is argued and demonstrated that techniques which have low resolutions such as microscopy will measure quite different kinetics: at short times a power of 14 as for wetting liquids and then a sudden equilibration as reported in these experiments. It is also argued on the basis of steric exclusion, and quantified in the disjoining pressure, that the behavior returns to that of wetting liquids when the polymer molecular weight becomes very high, as also observed in the experiments. Examples of how these features can find practical applications, and hence, the importance of use of polymers as additives are given. PMID- 16035798 TI - Spectral features of LO phonon sidebands in luminescence of free excitons in GaN. AB - In the paper a combined experimental and theoretical investigation of the longitudinal optical phonon sidebands (PSBs) in the luminescence of free excitons in GaN at moderately high temperatures was reported. The spectral features, including line broadening, shift, and asymmetry of the one- and two-phonon PSBs, were revealed both experimentally and theoretically. It is found that the linewidth of the one-phonon PSB is surprisingly always larger than that of the two-phonon PSB in the interested temperature range. Moreover, the thermal broadening rates of the one- and two-phonon PSBs are considerably different. We adopted the Segall-Mahan theory [B. Segall and G. D. Mahan, Phys. Rev. 171, 935 (1968)] to compute the PSB spectra of the free excitons in GaN. Only one adjustable parameter, the effective mass of the holes, was used in the calculations. For the one-phonon PSB, an excellent agreement between theory and experiment is achieved when an adequate effective mass of the holes was used. PMID- 16035799 TI - Rainbow scattering of CO and N2 from LiF(001). AB - The angular intensity distributions of CO and N(2) molecules scattered from a LiF(001) surface have been measured as functions of surface temperature, incident translational energy, and incident azimuthal direction affecting surface corrugation at a high resolution. Although both molecules have the same molecular mass and linear structure, only the CO molecule shows a rainbow feature in its scattering pattern, while the N(2) molecule shows a single peak distribution. From the comparisons of the obtained results with the calculated predictions based on the newly developed classical theory of the ellipsoid-washboard model, the differences in scattering distribution are attributed to the effects of molecular anisotropy and center-of-mass position. With an increase in the extent of the molecular anisotropy such as that of N(2) and CO as compared with rare-gas atoms, the summation of several scattering distributions depending on molecular orientation results in smearing the rainbow scattering on the corrugated surface. This smearing effect, however, attenuates when center-of-mass position deviates from the molecular center, as that for CO. PMID- 16035800 TI - Molecular dynamics investigation of the structural properties of phosphatidylethanolamine lipid bilayers. AB - We report a 14 ns microcanonical (NVE) molecular dynamics simulation of a fully hydrated bilayer of 1-stearoyl-2-oleoyl-phosphatidyethanolamine. This study describes the structure of the bilayer in terms of NMR order parameters and radial distribution functions, and compares them to experimental results and simulations of other lipids. A focus of this work is the characterization of the lipid-water interface, particularly the hydrogen bonding network of the phosphatidylethanolamine (PE) headgroups. We find that hydrogen bonding between the primary amine and phosphate groups has a pronounced effect on the structure of PE relative to phosphatidylcholine, and is evident in, for example, the P-N radial distribution functions. PMID- 16035801 TI - Molecular dynamics investigation of dynamical properties of phosphatidylethanolamine lipid bilayers. AB - We describe the dynamic behavior of a 1-stearoyl-2-oleoyl phosphatidylethanolamine (SOPE) bilayer from a 20 ns molecular dynamics simulation. The dynamics of individual molecules are characterized in terms of (2)H spin-lattice relaxation rates, nuclear overhauser enhancement spectroscopy (NOESY) cross-relaxation rates, and lateral diffusion coefficients. Additionally, we describe the dynamics of hydrogen bonding through an analysis of hydrogen bond lifetimes and the time evolution of clusters of hydrogen bonded lipids. The simulated trajectory is shown to be consistent with experimental measures of internal, intermolecular, and diffusive motion. Consistent with our analysis of SOPE structure in the companion paper, we see hydrogen bonding dominating the dynamics of the interface region. Comparison of (2)H T(1) relaxation rates for chain methylene segments in phosphatidylcholine and phosphatidylethanolamine bilayers indicates that slower motion resulting from hydrogen bonding extends at least three carbons into the hydrophobic core. NOESY cross-relaxation rates compare well with experimental values, indicating the observed hydrogen bonding dynamics are realistic. Calculated lateral diffusion rates (4 +/ -1 x 10(-8) cm(2)s) are comparable, though somewhat lower than, those determined by pulsed field gradient NMR methods. PMID- 16035802 TI - Multiscale coupling of mesoscopic- and atomistic-level lipid bilayer simulations. AB - A multiscale method is presented to bridge between the atomistic and mesoscopic membrane systems. The atomistic model in this case is the united atom dimyristoylphosphatidylcholine membrane system, although the method is completely general. Atomistic molecular dynamics provides the expansion modulus which is used to parametrize a mesoscopic elastic membrane model. The resulting elastic membrane model, including explicit mesoscopic solvent, shows appropriate static and dynamic undulation behaviors. Large membranes of approximately 100 nm in length can then be easily simulated using the mesoscopic membrane system. The critical feedback from the mesoscopic system back down to the atomistic-scale system is accomplished by bridging the stress (or surface tension) of a small region in the mesoscopic membrane to the corresponding atomistic membrane system. Because of long length-scale modes of membranes such as undulation and buckling, the local tension responds differently from the frame tension, when subjected to external perturbations. The effect of these membrane modes is shown for the stress response of a local membrane region and therefore the atomistic membrane system. In addition, certain equilibrium static and dynamic properties of stand alone and multiscale coupled systems are presented for several different membrane sizes. Although static properties such as two-dimensional pair-correlation function and order parameters show no noticeable discrepancy for the different systems, lipid self-diffusion and the rotational relaxation of lipid dipoles have a strong dependence on the membrane size (or long-wavelength membrane motions), which is properly modeled by the present multiscale method. PMID- 16035803 TI - Simulating the photoelectron spectra of rare-gas clusters. AB - Motivated by the recent experiments of the Swedish group [M. Tchaplyguine, R. R. Marinho, M. Gisselbrecht et al., J. Chem. Phys. 120, 345 (2004)], we simulate the photoelectron spectra of pure xenon and argon clusters. The clusters are modeled using molecular dynamics with Hartree-Fock-dispersion type pair potentials while the spectrum is calculated as the sum of final state energy shifts of the atoms ionized within the cluster relative to the isolated gas phase ion. A self consistent polarization formalism is used. Since signal electrons must travel through the cluster to reach the detector, we have accounted for the attenuation of the signal intensity by integrating an exponentially decaying scattering expression over the geometry of the cluster. Several different approaches to determining the required electron mean free paths (as a function of electron kinetic energy) are considered. Our simulated spectra are compared to the experimental results. PMID- 16035804 TI - Local structure of a phase-separating binary mixture in a mesoporous glass matrix studied by small-angle neutron scattering. AB - The mesoscopic structure of the binary system isobutyric acid + heavy water (D(2)O) confined in a porous glass (controlled-pore silica glass, mean pore width ca. 10 nm) was studied by small-angle neutron scattering at off-critical compositions in a temperature range above and below the upper critical solution point. The scattering data were analyzed in terms of a structure factor model similar to that proposed by Formisano and Teixeira [Eur. Phys. J. E 1, 1 (2000)], but allowing for both Ornstein-Zernike-type composition fluctuations and domainlike structures in the microphase-separated state of the pore liquid. The results indicate that the phase separation in the pores is shifted by ca. 10 K and spread out in temperature. Microphase separation is pictured as a transition from partial segregation at high temperature, due to the strong preferential adsorption of water at the pore wall, to a tube or capsule configuration of the two phases at low temperatures, depending on the overall composition of the pore liquid. Results for samples in which the composition of the pore liquid can vary with temperature due to equilibration with extra-pore liquid are consistent with this picture. PMID- 16035805 TI - Comparative study of O2 dissociation on various metalloporphyrins. AB - We investigate O(2) interaction with various metalloporphyrins (MnP, FeP, CoP, and NiP) using ab initio calculations based on density-functional theory. We discuss the trends in the activation barriers for the O-O bond cleavage in relation to the geometric, vibrational, electronic, and energetic properties of the systems. Whether the lowest unoccupied molecular orbital-highest occupied molecular orbital (LUMO-HOMO) levels of the metalloporphyrins involve the corresponding metal centers depends on the d orbital occupancies of the metals. We found that activation barriers for the O(2) dissociation can be mainly determined from the LUMO-HOMO characters of the metalloporphyrins, and consequently the FeP is the best catalyst with respect to the O(2) interaction from adsorption to dissociation. PMID- 16035806 TI - A reflection absorption infrared spectroscopy and density-functional theory investigation of methanol dehydrogenation on Rh(111)V alloy surfaces. AB - The dehydrogenation reaction of methanol on a Rh(111) surface, a Rh(111)V subsurface alloy, and on a Rh(111)V islands surface has been studied by thermal desorption spectroscopy, reflection absorption infrared spectroscopy, and density functional theory calculations. The full monolayer of methanol forms a structure with a special geometry with methanol rows, where two neighboring molecules have different oxygen-rhodium distances. They are close enough to form a H-bonded bilayer structure, with such a configuration, where every second methanol C-O bond is perpendicular to the surface on both Rh(111) and on the Rh(111)V subsurface alloy. The Rh(111)V subsurface alloy is slightly more reactive than the Rh(111) surface which is due to the changes in the electronic structure of the surface leading to slightly different methanol species on the surface. The Rh(111)V islands surface is the most reactive surface which is due to a new reaction mechanism that involves a methanol species stabilized up to about 245 K, partial opening of the methanol C-O bond, and dissociation of the product carbon monoxide. The latter two reactions also lead to a deactivation of the Rh(111)V islands surface. PMID- 16035807 TI - Interaction between benzenedithiolate and gold: classical force field for chemical bonding. AB - We have constructed a group of classical potentials based on ab initio density functional theory (DFT) calculations to describe the chemical bonding between benzenedithiolate (BDT) molecule and gold atoms, including bond stretching, bond angle bending, and dihedral angle torsion involved at the interface between the molecule and gold clusters. Three DFT functionals, local-density approximation (LDA), PBE0, and X3LYP, have been implemented to calculate single point energies (SPE) for a large number of molecular configurations of BDT-1, 2 Au complexes. The three DFT methods yield similar bonding curves. The variations of atomic charges from Mulliken population analysis within the molecule/metal complex versus different molecular configurations have been investigated in detail. We found that, except for bonded atoms in BDT-1, 2 Au complexes, the Mulliken partial charges of other atoms in BDT are quite stable, which significantly reduces the uncertainty in partial charge selections in classical molecular simulations. Molecular-dynamics (MD) simulations are performed to investigate the structure of BDT self-assembled monolayer (SAM) and the adsorption geometry of S adatoms on Au (111) surface. We found that the bond-stretching potential is the most dominant part in chemical bonding. Whereas the local bonding geometry of BDT molecular configuration may depend on the DFT functional used, the global packing structure of BDT SAM is quite independent of DFT functional, even though the uncertainty of some force-field parameters for chemical bonding can be as large as approximately 100%. This indicates that the intermolecular interactions play a dominant role in determining the BDT SAMs global packing structure. PMID- 16035808 TI - Photoinduced molecular reorientation dynamics in confined domains of Langmuir monolayers. AB - In a photoresponsive Langmuir monolayer comprised of smectic-C-like domains of mesogenic trans-azobenzene derivative embedded within an isotropic matrix of its cis isomer counterpart, several structurally differing circular droplets were irradiated with linearly polarized light. This report describes the structural rearrangements that occurred in these droplets upon illumination followed by Brewster angle microscopy analysis. Starting from initial well-characterized and symmetric states, final photoaligned situations were reached in which the azimuth angles of the rod-shaped elongated molecules were found to be perpendicular to the electric component of the excitation light. The dynamical aspects of the photoalignments, including their transient patterns, are captured by a theoretical model that couples a relaxational principle incorporating long-range elastic forces with a kinetic formalism presenting an anisotropic rate law. PMID- 16035809 TI - Structural determination of the low-coverage phase of Al on Si(001) surface. AB - The atomic structure of Al layer on Si(001)-(2 x 1) surface has been studied by coaxial impact collision ion scattering spectroscopy. When 0.5 monolayer (ML) of Al atoms are adsorbed on Si(001) at room temperature, it is found that Al adatoms are dimerized and Al ad-dimers are oriented parallel to the underlying Si dimers at the position of centering T3 site with a height of 1.02 Angstroms from the first layer of Si(001). The bond length of the Al dimer is 2.67 Angstroms. With increasing Al coverage up to one ML, Al ad-dimers still occupied near T3 site and the next favorable site is near HH site. PMID- 16035810 TI - Effect of chain length and asymmetry on material properties of bilayer membranes. AB - Dissipative particle dynamics is used to extract the material parameters (bending and area stretch moduli) of a bilayer membrane patch. Some experiments indicate that the area stretch modulus of lipid vesicles varies little as the chain length of the lipids composing the bilayer increases. Here we show that making the interactions between the hydrophilic head groups of the model amphiphiles proportional to the hydrophobic tail length reproduces the above result for the area stretch modulus. We also show that the area stretch modulus of bilayers composed of amphiphiles with the same number of tail beads but with asymmetric chains is less than that of bilayers with symmetric chains. The effects on the bilayer density and lateral stress profiles of changes to the amphiphile architecture are also presented. PMID- 16035811 TI - Asymmetry of lipid bilayers induced by monovalent salt: atomistic molecular dynamics study. AB - Interactions between salt ions and lipid components of biological membranes are essential for the structure, stability, and functions of the membranes. The specific ionic composition of aqueous buffers inside and outside of the cell is known to differ considerably. To model such a situation we perform atomistic molecular-dynamics (MD) simulations of a single-component phosphatidylcholine lipid bilayer which separates two aqueous reservoirs with and without NaCl salt. To implement the difference in electrolyte composition near two membrane sides, a double bilayer setup (i.e., two bilayers in a simulation box) is employed. It turns out that monovalent salt, being in contact with one leaflet only, induces a pronounced asymmetry in the structural, electrostatic, and dynamical properties of bilayer leaflets after 50 ns of MD simulations. Binding of sodium ions to the carbonyl region of the leaflet which is in contact with salt results in the formation of "Na-lipids" complexes and, correspondingly, reduces mobility of lipids of this leaflet. In turn, attractive interactions of chloride ions (mainly located in the aqueous phase close to the water-lipid interface) with choline lipid groups lead to a substantial (more vertical) reorientation of postphatidylcholine headgroups of the leaflet adjoined to salt. The difference in headgroup orientation on two sides of a bilayer, being coupled with salt-induced reorientation of water dipoles, leads to a notable asymmetry in the charge density profiles and electrostatic potentials of bilayer constitutes of the two leaflets. Although the overall charge density of the bilayer is found to be almost insensitive to the presence of salt, a slight asymmetry in the charge distribution between the two bilayer leaflets results in a nonzero potential difference of about 85 mV between the two water phases. Thus, a transmembrane potential of the order of the membrane potential in a cell can arise without ionic charge imbalance between two aqueous compartments. PMID- 16035812 TI - Brownian dynamics simulations of the self- and collective rotational diffusion coefficients of rigid long thin rods. AB - Recently a microscopic theory for the dynamics of suspensions of long thin rigid rods was presented, confirming and expanding the well-known theory by Doi and Edwards [The Theory of Polymer Dynamics (Clarendon, Oxford, 1986)] and Kuzuu [J. Phys. Soc. Jpn. 52, 3486 (1983)]. Here this theory is put to the test by comparing it against computer simulations. A Brownian dynamics simulation program was developed to follow the dynamics of the rods, with a length over a diameter ratio of 60, on the Smoluchowski time scale. The model accounts for excluded volume interactions between rods, but neglects hydrodynamic interactions. The self-rotational diffusion coefficients D(r)(phi) of the rods were calculated by standard methods and by a new, more efficient method based on calculating average restoring torques. Collective decay of orientational order was calculated by means of equilibrium and nonequilibrium simulations. Our results show that, for the currently accessible volume fractions, the decay times in both cases are virtually identical. Moreover, the observed decay of diffusion coefficients with volume fraction is much quicker than predicted by the theory, which is attributed to an oversimplification of dynamic correlations in the theory. PMID- 16035813 TI - Stationary flow, first passage times, and macroscopic Fick's first diffusion law: application to flow enhancement by particle trapping. AB - A generalized macroscopic Fick's first diffusion law is derived which describes steady-state particle flow between two baths explicitly as a function of the concentration gradient, acting as the thermodynamic driving force, times a functional of the first passage time. The latter is shown to be the ratio of the number of particles trapped between the baths and the first passage time. Particle trapping is shown to be a powerful mechanism by which flow can be enhanced. This is analyzed for two examples: a potential and an entropy trap. PMID- 16035814 TI - Temperature effects for isothermal polymer crystallization kinetics. AB - We adopt the cluster size distribution model to investigate the effect of temperature on homogeneous nucleation and crystal growth for isothermal polymer crystallization. The model includes the temperature effects of interfacial energy, nucleation rate, growth and dissociation rate coefficients, and equilibrium solubility. The time dependencies of polymer concentration, number and size of crystals, and crystallinity (in Avrami plots) are presented for different temperatures. The denucleation (Ostwald ripening effect) is also investigated by comparing moment and numerical solutions of the population balance equations. Agreement between the model results and temperature-sensitive experimental measurements for different polymer systems required strong temperature dependence for the crystal-melt interfacial energy. PMID- 16035815 TI - Exciton trapping at heterojunctions in polymer blends. AB - Optoelectronic devices made from semiconductor polymers often employ partially phase-separated binary polymer blends with "distributed heterojunctions" in the polymer film, and the migration of bulk excitons towards these heterojunctions crucially influences the device performance. Here, we investigate exciton migration in blend films of two polyfluorene derivatives. Localized exciplex states form in electron-hole capture at the heterojunction between the two polymers and these can be thermally excited to transfer to bulk excitons. Rapid radiative emission from these excitons can then allow efficient light-emitting diode operation. We show here that when these excitons migrate to another heterojunction site within their lifetime they are re-trapped at the interface and again form exciplex states or dissociate completely. We demonstrate that in polymer blend light-emitting diodes this can reduce the exciton population by more than 54% and can strongly influence the emission spectrum. We then analyze exciton re-trapping in detail using time-resolved photoluminescence spectroscopy on blends with different morphologies and find that for nanometer-scale phases exciton emission is completely suppressed. We show that the data agree well with a simple kinetic model which confirms the importance of the blend morphology for the exciton trapping efficiency. PMID- 16035816 TI - The effect of the range of interaction on the phase diagram of a globular protein. AB - Thermodynamic perturbation theory is applied to the model of globular proteins studied by ten Wolde and Frenkel [P. R. ten Wolde and D. Frenkel Science 77, 1975 (1997)] using computer simulation. It is found that the reported phase diagrams are accurately reproduced. The calculations show how the phase diagram can be tuned as a function of the length scale of the potential. PMID- 16035817 TI - Polypeptide foldings obtained with effective pair potentials. AB - We present a model of protein folding which is based on a potential function that describes the effective interaction between two amino acids (alanines, in this case). Our model is consistent with the formation of two important secondary structures, namely, an alpha-helix and a beta-ladder. In each case, we estimate the density of states using a random walk in energy space. This function allows the direct calculation of certain thermodynamic properties. By means of the configurational temperature, we also verify that the obtained polypeptides are in their native state. PMID- 16035818 TI - Heterogeneous dynamics of poly(vinyl acetate) far above Tg: a combined study by dielectric spectroscopy and quasielastic neutron scattering. AB - We have investigated the dynamics of poly(vinly acetate) using broadband dielectric spectroscopy (DS) covering over 14 decades in frequency up to 20 GHz and high-resolution quasielastic neutron-scattering (QENS) technique. The dielectric results have been interpreted in terms of the phenomenological Kohlrausch-Williams-Watts [G. Williams and D. C. Watts, Trans. Faraday Soc. 66, 80 (1970); F. Alvarez, A. Alegria, and J. Colmenero, Phys. Rev. B 47, 125 (1993)] description. Because of the wide frequency range covered by DS, it provides a precise determination of dynamics over a wide temperature range and it revealed a crossover in polymer dynamics at 387 K through different dielectric parameters, viz., characteristic times, asymmetric shape parameter, and dielectric strength. Moreover, shape parameter is found to be higher in comparison to other typical polymeric systems. The characteristic relaxation times observed by QENS displayed an anomalous dependence of momentum transfer, indicating the possible existence of heterogeneities in the system even at the high temperatures. In addition, spin lattice relaxation times, T(1), were found to be decoupled from dielectric characteristic times. Based on these results, a model was proposed to account for heterogeneities where we consider coexistence of different regions with standard polymeric behavior but with different characteristic times, leading to a distribution of relaxation times. The model is found to account for the anomalous behavior and an inherent shape parameter is found to account for the shape of alpha relaxation. This model is also found to predict the T variation of T(1) characteristic time scales at all temperatures. The origin of the heterogeneous domains is believed to lie in the microstructure of polymer chains. PMID- 16035819 TI - Coupling between lysozyme and glycerol dynamics: microscopic insights from molecular-dynamics simulations. AB - We explore possible molecular mechanisms behind the coupling of protein and solvent dynamics using atomistic molecular-dynamics simulations. For this purpose, we analyze the model protein lysozyme in glycerol, a well-known protein preserving agent. We find that the dynamics of the hydrogen bond network between the solvent molecules in the first shell and the surface residues of the protein controls the structural relaxation (dynamics) of the whole protein. Specifically, we find a power-law relationship between the relaxation time of the aforementioned hydrogen bond network and the structural relaxation time of the protein obtained from the incoherent intermediate scattering function. We demonstrate that the relationship between the dynamics of the hydrogen bonds and the dynamics of the protein appears also in the dynamic transition temperature of the protein. A study of the dynamics of glycerol as a function of the distance from the surface of the protein indicates that the viscosity seen by the protein is not the one of the bulk solvent. The presence of the protein suppresses the dynamics of the surrounding solvent. This implies that the protein sees an effective viscosity higher than the one of the bulk solvent. We also found significant differences in the dynamics of surface and core residues of the protein. The former is found to follow the dynamics of the solvent more closely than the latter. These results allowed us to propose a molecular mechanism for the coupling of the solvent-protein dynamics. PMID- 16035820 TI - Mixtures of charged colloid and neutral polymer: influence of electrostatic interactions on demixing and interfacial tension. AB - The equilibrium phase behavior of a binary mixture of charged colloids and neutral, nonadsorbing polymers is studied within free-volume theory. A model mixture of charged hard-sphere macroions and ideal, coarse-grained, effective sphere polymers is mapped first onto a binary hard-sphere mixture with nonadditive diameters and then onto an effective Asakura-Oosawa model [S. Asakura and F. Oosawa, J. Chem. Phys. 22, 1255 (1954)]. The effective model is defined by a single dimensionless parameter-the ratio of the polymer diameter to the effective colloid diameter. For high salt-to-counterion concentration ratios, a free-volume approximation for the free energy is used to compute the fluid phase diagram, which describes demixing into colloid-rich (liquid) and colloid-poor (vapor) phases. Increasing the range of electrostatic interactions shifts the demixing binodal toward higher polymer concentration, stabilizing the mixture. The enhanced stability is attributed to a weakening of polymer depletion-induced attraction between electrostatically repelling macroions. Comparison with predictions of density-functional theory reveals a corresponding increase in the liquid-vapor interfacial tension. The predicted trends in phase stability are consistent with observed behavior of protein-polysaccharide mixtures in food colloids. PMID- 16035821 TI - The elasticity of alpha-helices. AB - The elasticity of alpha-helices is examined using equilibrium molecular-dynamics simulations. From the statistics of curvatures and twists, we compute the elastic moduli of several representative alpha-helices, both in the presence and absence of aqueous solvent. We discover that the bending modulus (persistence length) of the helices is independent of the amino-acid sequence, although helices in water are slightly softer than in vacuum. The response of the helices under the action of an external force is also computed and compared with continuum mechanics predictions. Within the time scale of our simulation, we show that the properties of alpha-helices are well reproduced by an elastic and isotropic rod. The persistence length (bending modulus) of most alpha-helices in water or vacuum is approximately 100 nm, roughly twice that of DNA. PMID- 16035822 TI - Colloidal aggregation in polymer blends. AB - We consider here a low-density assembly of colloidal particles immersed in a critical polymer mixture of two chemically incompatible polymers. We assume that, close to the critical point of the free mixture, the colloids prefer to be surrounded by one polymer (critical adsorption). As result, one is assisted to a reversible colloidal aggregation in the nonpreferred phase, due the existence of a long-range attractive Casimir force between particles. This aggregation is a phase transition driving the colloidal system from dilute to dense phases, as the usual gas-liquid transition. We are interested in a quantitative investigation of the phase diagram of the immersed colloids. We suppose that the positions of particles are disordered, and the disorder is quenched and follows a Gaussian distribution. To apprehend the problem, use is made of the standard phi(4) theory, where the field phi represents the composition fluctuation (order parameter), combined with the standard cumulant method. First, we derive the expression of the effective free energy of colloids and show that this is of Flory-Huggins type. Second, we find that the interaction parameter u between colloids is simply a linear combination of the isotherm compressibility and specific heat of the free mixture. Third, with the help of the derived effective free energy, we determine the complete shape of the phase diagram (binodal and spinodal) in the (Psi,u) plane, with Psi as the volume fraction of immersed colloids. The continuous "gas-liquid" transition occurs at some critical point K of coordinates (Psi(c) = 0.5,u(c) = 2). Finally, we emphasize that the present work is a natural extension of that, relative to simple liquid mixtures incorporating colloids. PMID- 16035823 TI - Precise measurement of the self-diffusion coefficient for poly(ethylene glycol) in aqueous solution using uniform oligomers. AB - Uniform poly(ethylene glycol) (PEG) oligomers, with a degree of polymerization n=1-40, were separated by preparative supercritical fluid chromatography from commercial monodispersed samples. Diffusion coefficients, D, for separated uniform PEG oligomers were measured in dilute solutions of deuterium oxide (D(2)O) at 30 degrees C, using pulsed-field gradient nuclear magnetic resonance. The measured D for each molecular weight was extrapolated to infinite dilution. Diffusion coefficients obtained at infinite dilution follow the scaling behavior of Zimm-type diffusion, even in the lower molecular weight range. Molecular dynamics simulations for PEG in H(2)O also showed this scaling behavior, and reproduced close hydrodynamic interactions between PEG and water. These findings suggest that diffusion of PEG in water is dominated by hydrodynamic interaction over a wide molecular weight range, including at low molecular weights around 1000. PMID- 16035826 TI - Two new double Rydberg anions plus access to excited states of neutral Rydberg radicals via anion photoelectron spectroscopy. AB - We have observed and characterized two new double Rydberg anions N6H19- and N7H22 through their anion photoelectron spectra. The vertical detachment energies of these anions were found to be 0.443 and 0.438 eV, respectively. In addition, for three of the seven double Rydberg anions now known, we measured photodetachment transitions not only to the ground electronic states of their corresponding neutral Rydberg radicals but also to their first electronically excited states. In each spectrum, the energy spacing between the resulting peaks provided the ground-to-first electronically excited-state transition energy for the double Rydberg anion's corresponding neutral Rydberg radical. For the radicals, N4H13, N5H16, and N6H19, the spacings were found to be 0.83, 0.70, and 0.67 eV, respectively. These values are in excellent agreement with ground-to-first excited-state transition energies measured in absorption for the same neutral Rydberg radicals by Fuke and co-workers [Eur. Phys. J. D 9, 309 (1999); J. Phys. Chem. A 106, 5242 (2002).] The duplication of this neutral Rydberg property by photodetachment of double Rydberg anions further confirms that double Rydberg anions are indeed the negative ions of their corresponding neutral Rydberg molecules and cluster-like systems. PMID- 16035827 TI - Density functional theory study of indirect nuclear spin-spin coupling constants with spin-orbit corrections. AB - This work outlines the calculation of indirect nuclear spin-spin coupling constants with spin-orbit corrections using density functional response theory. The nonrelativistic indirect nuclear spin-spin couplings are evaluated using the linear response method, whereas the relativistic spin-orbit corrections are computed using quadratic response theory. The formalism is applied to the homologous systems H2X (X=O,S,Se,Te) and XH4 (X=C,Si,Ge,Sn,Pb) to calculate the indirect nuclear spin-spin coupling constants between the protons. The results confirm that spin-orbit corrections are important for compounds of the H2X series, for which the electronic structure allows for an efficient coupling between the nuclei mediated by the spin-orbit interaction, whereas in the case of the XH4 series the opposite situation is encountered and the spin-orbit corrections are negligible for all compounds of this series. In addition we analyze the performance of the density functional theory in the calculations of nonrelativistic indirect nuclear spin-spin coupling constants. PMID- 16035828 TI - Implementation of the locally renormalized CCSD(T) approaches for arbitrary reference function. AB - Several new variants of the locally-renormalized coupled-cluster (CC) approaches that account for the effect of triples (LR-CCSD(T)) have been formulated and implemented for arbitrary reference states using the TENSOR CONTRACTION ENGINE functionality, enabling the automatic generation of an efficient parallel code. Deeply rooted in the recently derived numerator-denominator-connected (NDC) expansion for the ground-state energy [K. Kowalski and P. Piecuch, J. Chem. Phys. 122, 074107 (2005)], LR-CCSD(T) approximations use, in analogy to the completely renormalized CCSD(T) (CR-CCSD(T)) approach, the three-body moments in constructing the noniterative corrections to the energies obtained in CC calculations with singles and doubles (CCSD). In contrast to the CR-CCSD(T) method, the LR-CCSD(T) approaches discussed in this paper employ local denominators, which assure the additive separability of the energies in the noninteracting system limit when the localized occupied spin-orbitals are employed in the CCSD and LR-CCSD(T) calculations. As clearly demonstrated on several challenging examples, including breaking the bonds of the F2, N2, and CN molecules, the LR-CCSD(T) approaches are capable of providing a highly accurate description of the entire potential-energy surface (PES), while maintaining the characteristic N(7) scaling of the ubiquitous CCSD(T) approach. Moreover, as illustrated numerically for the ozone molecule, the LR-CCSD(T) approaches yield highly competitive values for a number of equilibrium properties including bond lengths, angles, and harmonic frequencies. PMID- 16035829 TI - The electronic structure of vanadium carbide, VC. AB - Within an energy range of 2.4 eV, we have explored 29 of the 36 states of the diatomic molecule VC that arise from the atoms in their ground state, V(4s23d3;4F)+C(2s2 2p2;3P). We use multireference methods with large atomic natural orbital basis sets. The ground state is of 2Delta symmetry with the first two excited states, 4Delta and 2Sigma+, located 4.2 and 7.0 kcal/mol above the X state. All the states examined in this work are relatively strongly bound and show significant charge transfer from V to C. The binding energy of the X 2Delta state is estimated to be 95.3 kcal/mol in good agreement with the experimental value. PMID- 16035830 TI - First principles investigation of chromium carbide, CrC. AB - We have investigated the electronic structure of 14 states of the experimentally unknown diatomic molecule chromium carbide, CrC, using standard multireference configuration interaction methods and high quality basis sets. We report potential curves, binding energies, and a number of spectroscopic parameters. The ground state of CrC, X 3Sigma-, displays triple-bond character with a binding energy of D(e)=89 kcal/mol and an internuclear separation of r(e)=1.63 A. The first excited state (1 5Sigma-) lies 9.2 kcal/mol higher. All the states studied are fairly ionic, featuring an electron transfer of 0.3-0.5e- from the metal atom to the carbon atom. PMID- 16035831 TI - Analysis of the nuclear-electronic orbital method for model hydrogen transfer systems. AB - Fundamental issues associated with the application of the nuclear-electronic orbital (NEO) approach to hydrogen transfer systems are addressed. In the NEO approach, specified nuclei are treated quantum mechanically on the same level as the electrons, and mixed nuclear-electronic wavefunctions are calculated with molecular orbital methods. The positions of the nuclear basis function centers are optimized variationally. In the application of the NEO approach to hydrogen transfer systems, the hydrogen nuclei and all electrons are treated quantum mechanically. Within the NEO framework, the transferring hydrogen atom can be represented by two basis function centers to allow delocalization of the proton vibrational wavefunction. In this paper, the NEO approach is applied to the [He-H He]+ and [He-H-He]++ model systems. Analyses of technical issues pertaining to flexibility of the basis set to describe both single and double well proton potential energy surfaces, linear dependency of the hydrogen basis functions, multiple minima in the basis function center optimization, convergence of the number of hydrogen basis function centers, and basis set superposition error are presented. The accuracy of the NEO approach is tested by comparison to grid calculations for these model systems. PMID- 16035832 TI - Interdimensional degeneracies in van der Waals clusters and quantum Monte Carlo computation of rovibrational states. AB - Quantum Monte Carlo estimates of the spectrum of rotationally invariant states of noble gas clusters suggest interdimensional degeneracy in N-1 and N+1 spatial dimensions. We derive this property by mapping the Schrodinger eigenvalue problem onto an eigenvalue equation in which D appears as a continuous variable. We discuss implications for quantum Monte Carlo and dimensional scaling methods. PMID- 16035833 TI - Anharmonic midinfrared vibrational spectra of benzoic acid monomer and dimer. AB - Anharmonic vibrational calculations for the benzoic acid monomer and dimer in the mid-IR regime (500-1800 cm(-1)) are reported. Harmonic frequencies and intensities are obtained at the DFT/B3LYP level of theory employing D95(d,p) and cc-pVTZ basis sets. Anharmonic corrections obtained from standard perturbation theory lead to redshifts of 1%-3%. In almost all cases, the resulting frequencies deviate by less than 1% from previous measurements [Bakker et al., J. Chem. Phys. 119, 11180 (2003)]. Calculated intensities are in qualitative agreement with the absorption experiment, with the cc-pVTZ values being superior to the D95(d,p) ones for a few modes of the dimer. The antisymmetric out-of-plane bending mode of the dimer, which is strongly blueshifted with respect to the monomer frequency, represents a remarkable exception: The harmonic frequencies obtained for the two basis sets differ notably from each other, and the anharmonically corrected frequencies deviate from the experimental value by 8% [D95(d,p)] or 3% (cc-pVTZ). Nonperturbative calculations in reduced dimensionality reveal that the relatively small total anharmonic shift (few tens of cm(-1)) comprises of partly much larger contributions (few hundreds of cm(-1)) which are mostly canceling each other. Many of the individual anharmonic couplings are beyond the validity of second order perturbation theory based on cubic and semidiagonal quartic force constants only. This emphasizes the need for high-dimensional, nonperturbative anharmonic calculations at high quantum-chemical level when accurate frequencies of H-atom vibrations in double hydrogen bonds are sought for. PMID- 16035834 TI - Theory of the photodissociation of ozone in the Hartley continuum: potential energy surfaces, conical intersections, and photodissociation dynamics. AB - Ab initio potential energy and transition dipole moment surfaces are presented for the five lowest singlet even symmetry electronic states of ozone. The surfaces are calculated using the complete active space self consistent field method followed by contracted multireference configuration interaction (MRCI) calculations. A slightly reduced augmented correlation consistent valence triple zeta orbital basis set is used. The ground and excited state energies of the molecule have been computed at 9282 separate nuclear geometries. Cuts through the potential energy surfaces, which pass through the geometry of the minimum of the ground electronic state, show several closely avoided crossings. Close examination, and higher level calculations, very strongly suggests that some of these seemingly avoided crossings are in fact associated with non-symmetry related conical intersections. Diabatic potential energy and transition dipole moment surfaces are created from the computed ab initio adiabatic MRCI energies and transition dipole moments. The transition dipole moment connecting the ground electronic state to the diabatic B state surface is by far the strongest. Vibrational-rotational wavefunctions and energies are computed using the ground electronic state. The energy level separations compare well with experimentally determined values. The ground vibrational state wavefunction is then used, together with the diabatic B<--X transition dipole moment surface, to form an initial wavepacket. The analysis of the time-dependent quantum dynamics of this wavepacket provides the total and partial photodissociation cross sections for the system. Both the total absorption cross section and the predicted product quantum state distributions compare well with experimental observations. A discussion is also given as to how the observed alternation in product diatom rotational state populations might be explained. PMID- 16035835 TI - Photoelectron trapping in N2O 7sigma-->ksigma resonant ionization. AB - Vibrationally resolved photoelectron spectroscopy of the N2O+(A 2Sigma+) state is used to compare the dependence of the photoelectron dynamics on molecular geometry for two shape resonances in the same ionization channel. Spectra are acquired over the photon energy range of 18< or =hv< or =55 eV. There are three single-channel resonances in this range, two in the 7sigma-->ksigma channel and one in the 7sigma-->kpi channel. Vibrational branching ratio curves are determined by measuring vibrationally resolved photoelectron spectra as a function of photon energy, and theoretical branching ratio curves are generated via Schwinger variational scattering calculations. In the region 30< or =hv< or =40 eV, there are two shape resonances (ksigma and kpi). The ksigma ionization resonance is clearly visible in vibrationally resolved measurements at hv=35 eV, even though the total cross section in this channel is dwarfed by the cross section in the degenerate, more slowly varying 7sigma-->kpi channel. This ksigma resonance is manifested in non-Franck-Condon behavior in the approximately antisymmetric v3 stretching mode, but it is not visible in the branching ratio curve for the approximately symmetric v1 stretch. The behavior of the 35-eV ksigma resonance is compared to a previously studied N2O 7sigma-->ksigma shape resonance at lower energy. The mode sensitivity of the 35-eV ksigma resonance is the opposite of what was observed for the lower-energy resonance. The contrasting mode-specific behavior observed for the high- and low-energy 7sigma-->ksigma resonances can be explained on the basis of the "approximate" symmetry of the quasibound photoelectron resonant wave function, and the contrasting behavior reflects differences in the continuum electron trapping. An examination of the geometry dependence of the photoelectron dipole matrix elements shows that the ksigma resonances have qualitatively different dependences on the individual bond lengths. The low-energy resonance is influenced only by changes in the end-to-end length of the molecule, whereas the higher-energy resonance depends on the individual N-N and N-O bond lengths. Branching ratios are determined for several vibrational levels, including the symmetry-forbidden bending mode, and all of the observed behavior is explained in the context of an independent particle, Born Oppenheimer framework. PMID- 16035836 TI - Unimolecular rovibrational bound and resonance states for large angular momentum: J=20 calculations for HO2. AB - We explore the calculation of unimolecular bound states and resonances for deep well species at large angular momentum using a Chebychev filter diagonalization scheme incorporating doubling of the autocorrelation function as presented recently by Neumaier and Mandelshtam [Phys. Rev. Lett. 86, 5031 (2001)]. The method has been employed to compute the challenging J=20 bound and resonance states for the HO2 system. The methodology has firstly been tested for J=2 in comparison with previous calculations, and then extended to J=20 using a parallel computing strategy. The quantum J-specific unimolecular dissociation rates for HO2-->H+O2 in the energy range from 2.114 to 2.596 eV have been reported for the first time, and comparisons with the results of Troe and co-workers [J. Chem. Phys. 113, 11019 (2000) Phys. Chem. Chem. Phys. 2, 631 (2000)] from statistical adiabatic channel method/classical trajectory calculations have been made. For most of the energies, the reported statistical adiabatic channel method/classical trajectory rate constants agree well with the average of the fluctuating quantum mechanical rates. Near the dissociation threshold, quantum rates fluctuate more severely, but their average is still in agreement with the statistical adiabatic channel method/classical trajectory results. PMID- 16035837 TI - Accurate intermolecular ground-state potential-energy surfaces of the HCCH-He, Ne, and Ar van der Waals complexes. AB - Accurate ground-state intermolecular potential-energy surfaces are obtained for the HCCH-He, Ne, and Ar van der Waals complexes. The interaction energies are calculated at the coupled cluster singles and doubles including connected triple excitations level and fitted to analytic functions. For the three complexes we start with systematic basis set studies carried out at several intermolecular geometries, and using augmented correlation consistent polarized valence basis sets x-aug-cc-pVXZ (x=-,d; X=D,T,Q,5), also extended with a set of 3s3p2d1f1g midbond functions. The aug-cc-pVQZ-33211 surfaces of HCCH-He, Ne, and Ar complexes are characterized by absolute minima of -24.22, -50.20, and -122.17 cm( 1) at distances R between the rare-gas atom and the HCCH centers of mass of 4.35, 3.95, and 3.99 A, respectively; and at angles between the vector R and the HCCH main symmetry axis of 0 degrees , 43.3 degrees , and 60.6 degrees . The results are compared and considerably improve those previously available. PMID- 16035838 TI - The planar-to-tubular structural transition in boron clusters from optical absorption. AB - The optical response of the lowest-energy isomers of the B20 family is calculated using time-dependent density-functional theory within a real-space, real-time scheme. Significant differences are found among the absorption spectra of the clusters studied. We show that these differences can be easily related to changes in the overall geometry. Optical spectroscopy is thus an efficient tool to characterize the planar-to-tubular structural transition, known to be present in these boron-based systems. PMID- 16035839 TI - Scattering lengths for Li-Cs, Na-Cs, K-Cs, and Rb-Cs ultracold collisions. AB - We present triplet and singlet scattering lengths for X--133Cs collisions (X=6,7Li, 23Na, 39,40,41K, and 85,87Rb). We consider the short-range potential energy curves based on high-quality ab initio calculations and the long-range potential described by accurate dispersion coefficients available in the literature. Using a Fermi function to smoothly connect the short- and long-range terms we construct the potential for all R and evaluate the scattering length for each mixed species X--Cs. In particular, we predict that the cross section for inelastic processes between 23Na and 133Cs and between 40K and 133Cs will be small as compared with the respective elastic cross sections. PMID- 16035840 TI - Cavity ring-down spectroscopy and vibronic activity of benzo[ghi]perylene. AB - Gas-phase cavity ring-down spectroscopy of jet-cooled benzo[ghi]perylene (C22H12) in the 26 950-28 600-cm(-1) spectral range is reported for the first time. This study is part of our extensive laboratory astrophysics program for the study of interstellar polycyclic aromatic hydrocarbons. The observed spectrum shows an intermediate level structure and significant broadening and is associated with the vibronically coupled S1(1A1)<--S0(1A1) and S2(1B1)<--S0(1A1) electronic transitions. Time-dependent density-functional calculations were performed to calculate the energetics, vibrational frequencies, and normal coordinates of the S1 and S2 states. A simple vibronic model was employed to account for the vibronic interaction between the vibronic levels of the S1 and S2 states. The calculated vibronic spectrum is found to be in good agreement with the experimental spectrum. PMID- 16035841 TI - The ground and two lowest-lying singlet excited electronic states of copper hydroxide (CuOH). AB - Various ab initio methods, including self-consistent field (SCF), configuration interaction, coupled cluster (CC), and complete-active-space SCF (CASSCF), have been employed to study the electronic structure of copper hydroxide (CuOH). Geometries, total energies, dipole moments, harmonic vibrational frequencies, and zero-point vibrational energies are reported for the linear 1Sigma+ and 1Pi stationary points, and for the bent ground-state X 1A', and excited-states 2 1A' and 1 1A". Six different basis sets have been used in the study, Wachters/DZP being the smallest and QZVPP being the largest. The ground- and excited-state bending modes present imaginary frequencies for the linear stationary points, indicating that bent structures are more favorable. The effects of relativity for CuOH are important and have been considered using the Douglas-Kroll approach with cc-pVTZ/cc-pVTZ_DK and cc-pVQZ/cc-pVQZ_DK basis sets. The bent ground and two lowest-lying singlet excited states of the CuOH molecule are indeed energetically more stable than the corresponding linear structures. The optimized geometrical parameters for the X 1A' and 1 1A" states agree fairly well with available experimental values. However, the 2 1A' structure and rotational constants are in poor agreement with experiment, and we suggest that the latter are in error. The predicted adiabatic excitation energies are also inconsistent with the experimental values of 45.5 kcal mol(-1) for the 2 1A' state and 52.6 kcal mol( 1) for the 1 1A" state. The theoretical CC and CASSCF methods show lower adiabatic excitation energies for the 1 1A" state (53.1 kcal mol(-1)) than those for the corresponding 2 1A' state (57.6 kcal mol(-1)), suggesting that the 1 1A" state might be the first singlet excited state while the 2 1A' state might be the second singlet excited state. PMID- 16035842 TI - Dispersed fluorescence spectroscopy of jet-cooled HCF and DCF: Vibrational structure of the X 1A' state. AB - We recorded dispersed fluorescence (DF) spectra following excitation of the pure bending levels 2(0) (n) and the combination states 1(0) (1)2(0) (n) and 2(0) (n)3(0) (1) in the A 1A"<--X 1A' system of HCF and DCF. Spectra were measured with a 0.3 m spectrograph equipped with a gated intensified charge coupled device (CCD) detector and obtained under jet-cooled conditions using a pulsed discharge source. The DF spectra reveal rich detail concerning the vibrational structure of the X state up to 10 000 cm(-1). For HCF, resonances among the nearly degenerate levels 1(1)2n, 2n+13(1), and 2n+2 produce a polyadlike structure in the spectrum, and the usual effective spectroscopic Hamiltonian (Dunham expansion) poorly reproduces the experimental term energies. In contrast, this Hamiltonian works well for the term energies of DCF. Density functional calculations of the ground state vibrational frequencies were performed; the results are in excellent agreement with the experimentally derived vibrational parameters. The search for perturbations involving the low-lying a 3A" state is described. PMID- 16035843 TI - On the existence and lifetimes of Cu2+ complexes with water, ammonia, and hydrogen cyanide. AB - High-level ab initio calculations have been carried out to evaluate the lifetimes of complexes formed by the association of Cu2+ to water, ammonia, and hydrogen cyanide. The corresponding binding energies were evaluated at the CCSD(T)6 311+G(3df,2pd) level of theory. The potential-energy curves corresponding to their dissociation into Cu+ + L+ (L=H2O, NH3, and HCN) were obtained at the CCSD(T)6-311+G(3df,2p) level on BHLYP6-311+G(d,p) optimized geometries. Lifetimes were calculated using the exterior complex scaling and the semiclassical WKB methods. Although all the complexes investigated are thermodynamically unstable with respect to their dissociation into Cu+ + L+ in a typical Coulomb explosion, the activation barrier is high enough to accommodate several vibrational resonances, with very large lifetimes. As a matter of fact, if the three complexes are produced in the lowest vibrational levels, they behave as totally bound (with almost infinite lifetime) species. PMID- 16035844 TI - Renner-Teller vibronic analysis for a tetra-atomic molecule. I. The effective Hamiltonian and matrix elements. AB - The effective vibronic Hamiltonian for a linear tetra-atomic molecule in a Pi state has been investigated. In addition to the usual vibrational and Renner Teller coupling terms, the bending mode anharmonicity, spin-orbit coupling, and Fermi resonance interactions have been added to the model. Terms in the Hamiltonian up to the fourth order are given explicitly for molecules of C(infinityupsilon) symmetry and simplifications for symmetric D(infinityh) molecules are discussed. The matrix elements for the HCCS free radical have been obtained and are used to analyze the observed ground-state levels of HCCS and DCCS in a companion paper. The Sears resonance vibronic interaction that couples levels with the selection rules DeltaK=+/-1, DeltaSigma=-/+1, and DeltaP=0 has also been studied and the matrix elements derived. The determinable combinations of signs for the major parameters in the model are discussed. PMID- 16035845 TI - Renner-Teller vibronic analysis for a tetra-atomic molecule. II. The ground state of the HCCS free radical. AB - We have studied the tetra-atomic Renner-Teller effect in the ground state of the jet-cooled HCCS and DCCS free radicals. The reactive intermediates were produced in a pulsed discharge jet using thiophene or deuterated thiophene precursors and the ground-state vibronic energies were measured using the single vibronic level emission spectroscopy technique. In HCCS, emission and fluorescence depletion experiments proved that some previously unassigned laser-induced fluorescence (LIF) features were anomalous 2Delta32-2Pi32 transitions, which gave information on several ground-state 2Delta32 levels. In DCCS, rotational mixing of the excited state v'=0(2Pi32) level with the 4(1)(1 2Sigma12) level allowed several ground-state 2Sigma levels to be observed in emission. In addition, we have shown that some previously unassigned weak LIF bands of DCCS are anomalous 2Pi12-2Pi32 transitions which have allowed us to accurately measure the ground-state spin orbit splitting (259.0 cm(-1)) for the first time. All of the expected ground state 2Pi32 levels and the majority of the 2Pi12 levels up to 1700 cm(-1) have been observed for both isotopomers. The data have been fitted by an effective Hamiltonian matrix treatment that included Renner-Teller, spin-orbit coupling, anharmonicity, and Fermi resonance effects for the two bending vibrations and the CS stretching mode. This has allowed an unusually complete description of the orbital angular momentum coupling effects, including the determination of the epsilon4, epsilon5, and epsilon45 terms for the first time. Subtle effects in both the LIF and emission spectra signal the presence of Sears resonances in both the ground and excited states of these radicals. PMID- 16035846 TI - Pulsed-field ionization electron spectroscopy and binding energies of alkali metal-dimethyl ether and -dimethoxyethane complexes. AB - Lithium and sodium complexes of dimethyl ether (DME) and dimethoxyethane (DXE) were produced by reactions of laser-vaporized metal atoms with organic vapors in a pulsed nozzle cluster source. The mono-ligand complexes were studied by photoionization and pulsed field ionization zero electron kinetic energy (ZEKE) spectroscopy. Vibrationally resolved ZEKE spectra were obtained for Li(DME), Na(DME) and Li(DXE) and a photoionization efficiency spectrum for Na(DXE). The ZEKE spectra were analyzed by comparing with the spectra of other metal-ether complexes and with electronic structure calculations and spectral simulations. Major vibrations measured for the M(DME) (M=Li,Na) ions were M-O and C-O stretches and M-O-C and C-O-C bends. These vibrations and additional O-Li-O and O C-C-O bends were observed for the Li(DXE) ion. The M(DME) complexes were in C2v symmetry with the metal atom binding to oxygen, whereas Li(DXE) was in a C2 ring configuration with the Li atom attaching to both oxygen atoms. Moreover, the ionization energies of these complexes were measured from the ZEKE or photoionization spectra and bond dissociation energies were derived from a thermodynamic cycle. PMID- 16035847 TI - Density-functional theory (hyper)polarizabilities of push-pull pi-conjugated systems: treatment of exact exchange and role of correlation. AB - The performance of the optimized effective potential procedure for exact exchange in calculating static electric-field response properties of push-pull pi conjugated systems has been studied, with an emphasis on NO2-(CH=CH)n-NH2 chains. Good agreement with Hartree-Fock dipole moments and (hyper)polarizabilities is obtained; particularly noteworthy is the chain length dependence for beta/n. Thus, the problem that conventional density-functional theory functionals dramatically overestimate these properties is largely solved, although there remains a significant correlation contribution that cannot be accounted for with current correlation functionals. PMID- 16035848 TI - Ab initio prediction of the infrared-absorption spectrum of the C2Cl radical. AB - The three lowest (1(2)A', 2(2)A', and 1(2)A") potential-energy surfaces of the C2Cl radical, correlating at linear geometries with 2Sigma+ and 2Pi states, have been studied ab initio using a large basis set and multireference configuration interaction techniques. The electronic ground state is confirmed to be bent with a very low barrier to linearity, due to the strong nonadiabatic electronic interactions taking place in this system. The rovibronic energy levels of the 12C12C35Cl isotopomer and the absolute absorption intensities at a temperature of 5 K have been calculated, to an upper limit of 2000 cm(-1), using diabatic potential-energy and dipole moment surfaces and a recently developed variational method. The resulting vibronic states arise from a strong mixture of all the three electronic components and their assignments are intrinsically ambiguous. PMID- 16035849 TI - The electronic structure of liquid water within density-functional theory. AB - In the last decade, computational studies of liquid water have mostly concentrated on ground-state properties. However, recent spectroscopic measurements have been used to infer the structure of water, and the interpretation of optical and x-ray spectra requires accurate theoretical models of excited electronic states, not only of the ground state. To this end, we investigate the electronic properties of water at ambient conditions using ab initio density-functional theory within the generalized gradient approximation (DFT/GGA), focusing on the unoccupied subspace of Kohn-Sham eigenstates. We generate long (250 ps) classical trajectories for large supercells, up to 256 molecules, from which uncorrelated configurations of water molecules are extracted for use in DFT/GGA calculations of the electronic structure. We find that the density of occupied states of this molecular liquid is well described with 32-molecule supercells using a single k point (k=0) to approximate integration over the first Brillouin zone. However, the description of the unoccupied electronic density of states (u-EDOS) is sensitive to finite size effects. Small, 32-molecule supercell calculations, using the Gamma-point approximation, yield a spuriously isolated state above the Fermi level. Nevertheless, the more accurate u-EDOS of large, 256-molecule supercells may be reproduced using smaller supercells and increased k-point sampling. This indicates that the electronic structure of molecular liquids such as water is relatively insensitive to the long-range disorder in the molecular structure. These results have important implications for efficiently increasing the accuracy of spectral calculations for water and other molecular liquids. PMID- 16035850 TI - Emergence of the genuine Johari-Goldstein secondary relaxation in m-fluoroaniline after suppression of hydrogen-bond-induced clusters by elevating temperature and pressure. AB - The dielectric spectra of the glass former, m-fluoroaniline (m-FA), at ambient pressure show the presence of a secondary relaxation, which was identified in the literature as the universal Johari-Goldstein (JG) beta relaxation. However, published elastic neutron scattering and simulation data [D. Morineau, C. Alba Simionesco, M. C. Bellisent-Funel, and M. F. Lauthie, Europhys. Lett. 43, 195 (1998); D. Morineau and C. Alba-Simionesco, J. Chem. Phys. 109, 8494 (1998)] showed the presence of hydrogen-bond-induced clusters of limited size in m-FA at ambient pressure and temperature of the dielectric measurements. The observed secondary relaxation may originate from the hydrogen-bond-induced clusters. If so, it should not be identified with the JG beta relaxation that involves essentially all parts of the molecule and has certain characteristics [K. L. Ngai and M. Paluch, J. Chem. Phys. 120, 857 (2004)], but then arises the question of where is the supposedly universal JG beta relaxation in m-FA. To gain a better understanding and resolving the problem, we perform dielectric measurements at elevated pressures and temperatures to suppress the hydrogen-bond-induced clusters and find significant changes in the dielectric spectra. The secondary relaxation observed at ambient pressure in m-FA is suppressed, indicating that indeed it originates from the hydrogen-bond-induced clusters. The spectra of m-FA are transformed at high temperature and pressure to become similar to that of toluene. The new secondary relaxation that emerges in the spectra has properties of a genuine JG relaxation like in toluene. PMID- 16035851 TI - Multidimensional vibrational spectroscopy for tunneling processes in a dissipative environment. AB - Simulating tunneling processes as well as their observation are challenging problems for many areas. In this study, we consider a double-well potential system coupled to a heat bath with a linear-linear (LL) and square-linear (SL) system-bath interactions. The LL interaction leads to longitudinal (T1) and transversal (T2) homogeneous relaxations, whereas the SL interaction leads to the inhomogeneous dephasing (T2*) relaxation in the white noise limit with a rotating wave approximation. We discuss the dynamics of the double-well system under infrared (IR) laser excitations from a Gaussian-Markovian quantum Fokker-Planck equation approach, which was developed by generalizing Kubo's stochastic Liouville equation. Analytical expression of the Green function is obtained for a case of two-state-jump modulation by performing the Fourier-Laplace transformation. We then calculate a two-dimensional infrared signal, which is defined by the four-body correlation function of optical dipole, for various noise correlation time, system-bath coupling parameters, and temperatures. It is shown that the bath-induced vibrational excitation and relaxation dynamics between the tunneling splitting levels can be detected as the isolated off diagonal peaks in the third-order two-dimensional infrared (2D-IR) spectroscopy for a specific phase matching condition. Furthermore, this spectroscopy also allows us to directly evaluate the rate constants for tunneling reactions, which relates to the coherence between the splitting levels; it can be regarded as a novel technique for measuring chemical reaction rates. We depict the change of reaction rates as a function of system-bath coupling strength and a temperature through the 2D-IR signal. PMID- 16035852 TI - On the temperature and pressure dependences of cavities in the dielectric continuum picture. AB - Cavity size at equilibrium and its variations with thermodynamic conditions are studied in the dielectric continuum framework of solvents. By employing Gibbs' theory of dividing surfaces, the fluctuating cavity description of Kim [H.J. Kim, J. Chem. Phys. 105, 6818 (1996)] is extended to include effects related to the local solvent density inhomogeneity near the cavity. The resulting theory is applied to study cavity size variations with temperature and pressure in dipolar and nondipolar solvents. Model calculations show that the cavity size tends to increase with temperature along an isobar and decrease with pressure along an isotherm. PMID- 16035853 TI - Transport coefficients and dynamic properties of hydrogen sulfide from molecular simulation. AB - Molecular-dynamics simulation results on thermodynamic and transport properties of pure H2S under conditions of practical interest are presented. Our data are in very good quantitative agreement with the scarce experimental data and estimates on thermophysical properties of this substance. Our results serve as a test of the validity of the intermolecular potential used in the simulations as well as the consistency of the existing data in the studied range. New simulation data on thermal conductivity at low temperature as well as in supercritical states are also reported. Furthermore, we present a comparative analysis between the local order in the liquid phase of pure hydrogen sulfide and water, due to the molecular analogies between both substances, and its relation with the formation of H=S bonds. Our results indicate that under the same corresponding thermodynamic states, H2S is a much less structured substance, with a first solvation shell with a dodecahedral order instead of the tetrahedral order observed in water. PMID- 16035854 TI - Universal isotope effect in thermal diffusion of mixtures containing cyclohexane and cyclohexane-d12. AB - The Soret coefficients S(T) of the liquids acetone, benzene, benzene-d1, 1,3,5 benzene-d3, benzene-d5, benzene-13C6, benzene-d6, n-hexane, toluene, 1,2,3,4 tetrahydronaphthalene, isobutylbenzene, and 1,6-dibromohexane have been measured in protonated and perdeuterated cyclohexane by a transient holographic grating technique. It has been found that S(T) can be either positive or negative and even change its sign as a function of concentration. The isotope effect DeltaS(T)=-0.99 x 10(-3) K(-1), which is the change of S(T) after isotopic substitution of cyclohexane, neither depends on concentration nor on the nature of the mixing partner. Only in the case of the polar acetone is DeltaS(T) approximately 30% larger but still concentration independent. Based on the experimental findings, some general conclusions about the dependence of the Soret coefficient on molecular properties are drawn. PMID- 16035855 TI - The equation of state of hard hyperspheres in four and five dimensions. AB - The equation of state of hard hyperspheres in four and five dimensions is calculated from the value of the pair correlation function at contact, as determined by Monte Carlo simulations. These results are compared to equations of state obtained by molecular dynamics and theoretical approaches. In all cases the agreement is excellent. PMID- 16035856 TI - Composition and temperature dependence of cesium-borate glasses by molecular dynamics. AB - The structural aspects of xCs2O-(1-x)B2O3 glasses have been investigated by molecular dynamics as functions of Cs2O content (x=0.2, 0.3, and 0.4) and temperature (T=300 and 1250 K). The tetrahedral (BO4-) and triangular (BO3,BO2O-, and BOO2 (2-)) short-range order borate units were found to be the structure building entities of the simulated glasses [O=bridging oxygen (BO) and O =nonbridging oxygen (NBO) atom]. The increase of Cs2O content results in the progressive increase of the NBO-containing triangle population at the expense of the BO4- tetrahedral units. The same effect is caused by temperature increase at a fixed Cs2O content, and this was associated with the "fragile" characteristics of alkali borate glasses. A comparison of simulated Cs and Li borates showed very similar structures at x=0.2, but dissimilar ones when the alkali content exceeds this composition. In particular, for x>0.2 Cs borates exhibit a preference for NBO formation relative to Li borates. Differences in the microstructure of sites hosting Cs ions were found, and this permits their classification into bridging (b type) and nonbridging type (nb type) of sites. b-type sites consist exclusively of BO atoms, while both BO and NBO atoms participate in nb-type sites. These differences in Cs-site local bonding characteristics were found to be reflected on the Cs-O(site) vibration frequencies. Also, the computed Cs-O vibrational responses for simulated Cs borates were found to compare well with experimental far-infrared spectra. PMID- 16035857 TI - Binding energies of hydrogen molecules to isoreticular metal-organic framework materials. AB - Recently, several novel isoreticular metal-organic framework (IRMOF) structures have been fabricated and tested for hydrogen storage applications. To improve our understanding of these materials, and to promote quantitative calculations and simulations, the binding energies of hydrogen molecules to the MOF have been studied. High-quality second-order Moller-Plesset (MP2) calculations using the resolution of the identity approximation and the quadruple zeta QZVPP basis set were used. These calculations use terminated molecular fragments from the MOF materials. For H2 on the zinc oxide corners, the MP2 binding energy using Zn4O(HCO2)6 molecule is 6.28 kJ/mol. For H2 on the linkers, the binding energy is calculated using lithium-terminated molecular fragments. The MP2 results with coupled-cluster singles and doubles and noniterative triples method corrections and charge-transfer corrections are 4.16 kJ/mol for IRMOF-1, 4.72 kJ/mol for IRMOF-3, 4.86 kJ/mol for IRMOF-6, 4.54 kJ/mol for IRMOF-8, 5.50 and 4.90 kJ/mol for IRMOF-12, 4.87 and 4.84 kJ/mol for IRMOF-14, 5.42 kJ/mol for IRMOF-18, and 4.97 and 4.66 kJ/mol for IRMOF-993. The larger linkers are all able to bind multiple hydrogen molecules per side. The linkers of IRMOF-12, IRMOF-993, and IRMOF-14 can bind two to three, three, and four hydrogen molecules per side, respectively. In general, the larger linkers have the largest binding energies, and, together with the enhanced surface area available for binding, will provide increased hydrogen storage. We also find that adding up NH2 or CH3 groups to each linker can provide up to a 33% increase in the binding energy. PMID- 16035858 TI - Friction force microscopy as an alternative method to probe molecular interactions. AB - Friction force microscopy was applied to study protein-carbohydrate interactions that are important in many cellular recognition processes. The expression and structure of carbohydrates can be investigated using lectins as molecular probes since they recognize different types of sugar molecules. Lectins (concanavalin A and lentil lectin, recognizing mannose-type carbohydrates) were attached to the probing tip and carboxypeptidase Y (possessing complementary carbohydrates) was immobilized on a modified glass surface using microcontact printing. The results obtained from friction force maps and dependencies on the loading rate (measured in a physiological buffer) were divided in two distinct groups. The first group of results obtained for lectin-protein complexes was assigned to molecular recognition events, whereas the other including all control measurements was attributed to nonspecific interaction. All results presented here indicate that friction force microscopy can be successfully employed to study recognition processes. PMID- 16035859 TI - Lattice defects and magnetic ordering in plutonium oxides: a hybrid density functional-theory study of strongly correlated materials. AB - Experimental studies of actinide oxides are challenging, and conventional electronic structure calculations fail to qualitatively reproduce the scarce data. We employ a new generation of hybrid density functionals to model a defective plutonium dioxide lattice. The procedure is first tested against stoichiometric bulk PuO2 and Pu2O3, for which predictions agree well with experiment where known. The interstitial oxygen in PuO2.25 is found to be singly charged, consistent with experimental observations and contrary to the O2- previously proposed theoretically. PMID- 16035860 TI - Evaluation of sliding friction and contact mechanics of elastomers based on dynamic-mechanical analysis. AB - The paper presents a combined experimental and theoretical approach to the understanding of hysteresis and adhesion contributions to rubber friction on dry and lubricated rough surfaces. Based on a proper analysis of the temperature- and frequency-dependent behaviors of nonlinear viscoelastic materials such as filler reinforced elastomer materials, master curves for the viscoelastic moduli are constructed. It is shown that the classical williams-Landel-Ferry equation cannot be applied in its simple form, but needs the introduction of an energy term describing the temperature dependency of glassy polymer bridges, which transmit the forces within flocculated filler clusters. The activation energy for carbon black and silica-filled elastomers is compared based on two different evaluation methods. The obtained dynamic data are shown to be related to a different friction behavior of elastomers regarding the two filler systems. Theoretical predictions of the stationary frictional behavior of the systems are in fair agreement with the experimental friction data at low sliding velocities. It is found that the formulated adhesion plays a dominant role on rough dry surfaces within this range of velocities. PMID- 16035861 TI - Anomalous vibrational energy diffusion in carbon nanotubes. AB - We study the vibrational energy diffusion in single-walled carbon nanotubes by using the molecular-dynamics method. It is found that energy transports ballistically at low temperature and superdiffusively at room temperature. The velocity of energy transport along the axis in carbon nanotube at room temperature is about 0.10 A/fs. It is also found that energy transport in carbon nanotube is different from that one in one-dimensional carbon lattice with the same interaction potential. PMID- 16035862 TI - Anisotropic diffusion of n-butane and n-decane on a stepped metal surface. AB - The diffusion of single n-butane and n-decane molecules on a model stepped surface, Pt655, and on a corresponding flat surface, Pt111, is investigated using molecular-dynamics simulations and anisotropic united atom model. The surface step on Pt655 causes the alkane molecules to adsorb on the lower terrace in all trans conformations with their long molecular axes adjacent and parallel to the step edge, and to diffuse anisotropically along the surface step via a constant wiggly motion without rotation or marked deviation from the parallel adsorption configuration. At relatively high temperatures, the alkane molecules can temporarily break away from the step edge but cannot migrate across the step edge in either the downstair or upstair direction. In comparison with the diffusion on Pt111, the diffusivity of n-decane is reduced by the surface step but its diffusion barrier is hardly affected. In the case of the shorter n-butane, however, the surface step significantly reduces the diffusion energy barrier and gives rise to higher diffusion coefficients at lower temperatures. Important implications of the simulation results are discussed. PMID- 16035863 TI - Orientation-dependent potential of mean force for protein folding. AB - We present a solvent-implicit minimalistic model potential among the amino acid residues of proteins, obtained by using the known native structures [deposited in the Protein Data Bank (PDB)]. In this model, the amino acid side chains are represented by a single ellipsoidal site, defined by the group of atoms about the center of mass of the side chain. These ellipsoidal sites interact with other sites through an orientation-dependent interaction potential which we construct in the following fashion. First, the site-site potential of mean force (PMF) between heavy atoms is calculated [following F. Melo and E. Feytsman, J. Mol. Biol. 267, 207 (1997)] from statistics of their distance separation obtained from crystal structures. These site-site potentials are then used to calculate the distance and the orientation-dependent potential between side chains of all the amino acid residues (AAR). The distance and orientation dependencies show several interesting results. For example, we find that the PMF between two hydrophobic AARs, such as phenylalanine, is strongly attractive at short distances (after the obvious repulsive region at very short separation) and is characterized by a deep minimum, for specific orientations. For the interaction between two hydrophilic AARs, such a deep minimum is absent and in addition, the potential interestingly reveals the combined effect of polar (charge) and hydrophobic interactions among some of these AARs. The effectiveness of our potential has been tested by calculating the Z-scores for a large set of proteins. The calculated Z-scores show high negative values for most of them, signifying the success of the potential to identify the native structure from among a large number of its decoy states. PMID- 16035864 TI - Morphological variation in a toroid generated from a single polymer chain. AB - A single semiflexible polymer chain folds into a toroidal object under poor solvent conditions. In this study, we examined the morphological change in such a toroidal state as a function of the cross-sectional area and stiffness of the chain together with the surface energy, which characterizes the segmental interaction parameter. Changes in the thickness and outer/inner radius on a toroid are interpreted in terms of these parameters. Our theoretical expectation corresponds to the actual morphological changes in a single giant DNA molecule as observed by electron microscopy. PMID- 16035865 TI - Small-angle neutron scattering of dilute polystyrene chains at the protein limit of a colloid-polymer mixture. AB - The present work investigates the structure of large polystyrene (PS) chains in solutions of small poly(ethyl methacrylate) (PEMA) microgel particles in toluene. Toluene is a good solvent for the PS chains. The PEMA colloids have an outer radius of R=11 nm which is much smaller in size than the radius of gyration Rg=58 nm of the PS chains. The system is considered to represent the protein limit of polymer-colloid mixtures. Structural investigation is performed by small-angle neutron scattering (SANS) using an appropriate contrast matching. This could be achieved by dissolving fully deuterated PS chains (D8-PS) in solutions of hydrogenated PEMA colloids in hydrogenated toluene (H-toluene). It is first demonstrated that PEMA colloids are satisfactorily contrast matched for SANS in H toluene if the PEMA concentration does not exceed 200 g/l. Based on these findings, D8-PS is investigated by SANS in pure H-toluene and three different PEMA concentrations in H-toluene. The results indicate a drastic shrinking of D8 PS chain dimensions with increasing PEMA content. Comparison with model curves of star-branched Gaussian chains and Gaussian rings suggest a striking similarity of the respective density-density correlation of those models with the shrunken D8 PS chains. Along with this, a shrinking as large as 0.5 was estimated when the PEMA content reached 200 g/l. PMID- 16035866 TI - Spinodal for the solution-to-crystal phase transformation. AB - The formation of crystalline nuclei from solution has been shown for many systems to occur in two steps: the formation of quasidroplets of a disordered intermediate, followed by the nucleation of ordered crystalline embryos within these droplets. The rate of each step depends on a respective free-energy barrier and on the growth rate of its near-critical clusters. We address experimentally the relative significance of the free-energy barriers and the kinetic factors for the nucleation of crystals from solution using a model protein system. We show that crystal nucleation is 8-10 orders of magnitude slower than the nucleation of dense liquid droplets, i.e., the second step is rate determining. We show that at supersaturations of three or four k(B)T units, crystal nuclei of five, four, or three molecules transform into single-molecule nuclei, i.e., the significant nucleation barrier vanishes below the thermal energy of the molecules. We show that the main factor, which determines the rate of crystal nucleation, is the slow growth of the near-critical ordered clusters within the quasidroplets of the disordered intermediate. Analogous to the spinodal in supersaturated fluids, we define a solution-to-crystal spinodal from the transition to single-molecule crystalline nuclei. We show that heterogeneous nucleation centers accelerate nucleation not only because of the wettinglike effects that lower the nucleation barrier, as envisioned by classical theory, but by helping the kinetics of growth of the ordered crystalline embryos. PMID- 16035868 TI - Dynamics and rheology of immiscible polymer-liquid-crystal systems. AB - The morphology evolution in immiscible polymer-liquid-crystal systems is quite different from flexible polymer-polymer mixtures due to the anisotropic properties of liquid crystals. The dynamics and rheology of such system are discussed. A theoretical model is proposed to describe the dynamics of liquid crystal droplets in a flow field and the rheological properties of immiscible liquid-crystal/polymer mixtures. The deformation of liquid-crystal droplet is found to be greatly dependent on the interfacial properties of polymer-liquid crystal. The scaling relationships of interfacial contribution to the stresses are found to be quite different from the isotropic mixtures with droplet morphology. PMID- 16035867 TI - Reliable treatment of electrostatics in combined QM/MM simulation of macromolecules. AB - A robust approach for dealing with electrostatic interactions for spherical boundary conditions has been implemented in the QM/MM framework. The development was based on the generalized solvent boundary potential (GSBP) method proposed by Im et al. [J. Chem. Phys. 114, 2924 (2001)], and the specific implementation was applied to the self-consistent-charge density-functional tight-binding approach as the quantum mechanics (QM) level, although extension to other QM methods is straightforward. Compared to the popular stochastic boundary-condition scheme, the new protocol offers a balanced treatment between quantum mechanics/molecular mechanics (QM/MM) and MM/MM interactions; it also includes the effect of the bulk solvent and macromolecule atoms outside of the microscopic region at the Poisson Boltzmann level. The new method was illustrated with application to the enzyme human carbonic anhydrase II and compared to stochastic boundary-condition simulations using different electrostatic treatments. The GSBP-based QM/MM simulations were most consistent with available experimental data, while conventional stochastic boundary simulations yielded various artifacts depending on different electrostatic models. The results highlight the importance of carefully treating electrostatics in QM/MM simulations of biomolecules and suggest that the commonly used truncation schemes should be avoided in QM/MM simulations, especially in simulations that involve extensive conformational samplings. The development of the GSBP-based QM/MM protocol has opened up the exciting possibility of studying chemical events in very complex biomolecular systems in a multiscale framework. PMID- 16035869 TI - Sub-Tg dynamics in polycarbonate by neutron scattering and its relation with secondary gamma relaxation. AB - We have investigated the dynamics of phenylene rings in glassy bisphenol-A (BPA) polycarbonate (PC) by means of quasielastic neutron scattering. Taking advantage of selective deuteration of the samples, we have studied the incoherent scattering of hydrogens in phenylene rings on the one hand, and on the other hand the coherent quasielastic scattering of all the atoms in the sample. Two different types of neutron spectrometers, time of flight and backscattering, were used in order to cover a wide dynamic range, which extends from microscopic (approximately 10(-13) s) to mesoscopic (approximately 10(-9) s) times. Moreover, neutron-diffraction experiments with polarization analysis were carried out in order to characterize the structural features, and the relative coherent and incoherent contributions of the samples investigated. In contrast with previous studies of phenylene ring dynamics in BPA polysulfone performed by us also by neutron scattering, phenylene rings in BPA PC exhibit an "extra" motion in addition to those found for BPA polysulfone's phenylene rings. This extra motion of the rings in PC perfectly correlates with the main carbonate group motion followed by dielectric spectroscopy and allows us to (i) consistently interpret the PC's gamma relaxation in terms of two different motions; and (ii) experimentally confirm the relation between the motion of phenylene rings and carbonate groups within BPA PC formerly predicted by computational methods. PMID- 16035870 TI - A global investigation of phase equilibria using the perturbed-chain statistical associating-fluid-theory approach. AB - The recently developed perturbed-chain statistical-associating-fluid theory (PC SAFT) is investigated for a wide range of model parameters including the parameter m representing the chain length and the thermodynamic temperature T and pressure p. This approach is based upon the first-order thermodynamic perturbation theory for chain molecules developed by Wertheim [M. S. Wertheim, J. Stat. Phys. 35, 19 (1984); ibid. 42, 459 (1986)] and Chapman et al. [G. Jackson, W. G. Chapman, and K. E. Gubbins, Mol. Phys. 65, 1 (1988); W. G. Chapman, G. Jackson, and K. E. Gubbins, ibid. 65, 1057 (1988)] and includes dispersion interactions via the second-order perturbation theory of Barker and Henderson [J. A. Barker and D. Henderson, J. Chem. Phys. 47, 4714 (1967)]. We systematically study a hierarchy of models which are based on the PC-SAFT approach using analytical model calculations and Monte Carlo simulations. For one-component systems we find that the analytical model in contrast with the simulation results exhibits two phase-separation regions in addition to the common gas-liquid coexistence region: One phase separation occurs at high density and low temperature. The second demixing takes place at low density and high temperature where usually the ideal-gas phase is expected in the phase diagram. These phenomena, which are referred to as "liquid-liquid" and "gas-gas" equilibria, give rise to multiple critical points in one-component systems, as well as to critical end points and equilibria of three fluid phases, which can usually be found in multicomponent mixtures only. Furthermore, it is shown that the liquid liquid demixing in this model is not a consequence of a "softened" repulsive interaction as assumed in the theoretical derivation of the model. Experimental data for the melt density of polybutadiene with molecular mass Mw=45,000 gmol are correlated here using the PC-SAFT equation. It is shown that the discrepancies in modeling the polymer density at ambient temperature and high pressure can be traced back to the liquid-liquid phase separation predicted by the equation of state at low temperatures. This investigation provides a basis for understanding possible inaccuracies or even unexpected phase behavior which can occur in engineering applications of the PC-SAFT model aiming at predicting properties of macromolecular substances. PMID- 16035871 TI - Effect of chain stiffness and entanglements on the elastic behavior of end-linked elastomers. AB - The effect of chain stiffness and entanglements on the elastic behavior and microscopic structure of cross-linked polymer networks was studied using Monte Carlo simulations. We investigated the behavior of entangled and entanglement free networks at various degrees of chain stiffness and densities. Based on previous results that indicated that trapped entanglements prevent strain-induced order-disorder transitions in semiflexible chain networks, we prepared the entangled networks by end-linking the chains in very dilute conditions so as to minimize the extent of trapped entanglements. We also considered the entanglement free case by using a "diamond" structure. We found that the presence of even a very small amount of trapped entanglements is enough to prevent a discontinuous strain-induced transition to an ordered phase. In these mildly entangled networks, a nematiclike order is eventually attained at high extensions but the elastic response remains continuous and the cross-links remain uniformly distributed through the simulation box. The entanglement-free diamond networks on the other hand show discontinuities in their stress-strain data. Networks at higher densities exhibit a more stable ordered phase and show an unusual staircaselike stress-strain curve. This is the result of a stepwise extension mechanism in which the chains form ordered domains that exclude the cross-links. Extension is achieved by increasing the number of these ordered domains in the strain direction. Cross-links aggregate in the spaces between these ordered domains and form periodic bands. Each vertical upturn in the stress-strain data corresponds to the existence of an integer number of ordered domains. This stepwise elastic behavior is found to be similar to that exhibited by some tough natural materials. PMID- 16035872 TI - Local friction in polyolefin blends. AB - Processes on different length scales affect the dynamics of chain molecules. The friction experienced by a short chain segment depends on both small-scale chain properties and on the local environment of the segment. As a consequence, the (monomeric) friction coefficients of the two components of a binary polymer blend will, in general, differ from each other and from the friction coefficients of the corresponding melts. In this work, we investigate local friction in polyolefin blends with the aid of a small-scale simulation approach. The polymer chains, in united atom representation, are assumed to occupy the sites of a partially filled simple cubic lattice. The simulation focuses on short chain sections with straight backbones and enumerates all possible binary contacts and relative movements of such sections. By evaluating the exact enumeration results in conjunction with equations of state for the blends, we are able to make predictions about the variation of the friction coefficients with local chain architecture and thermodynamic state (temperature, pressure, and composition). We calculate relative values of friction coefficients at temperatures well above the glass transition for blends of PEP, an alternating copolymer of polyethylene and polypropylene, with polyethylene and polyisobutylene and for blends of polyethylene and atactic polypropylene. We also investigate a blend of PEP with head-to-head polypropylene and compare our results with experimental data. PMID- 16035874 TI - On the notion of "1/f noise" and data analysis for glassy water. PMID- 16035873 TI - Temperature dependence of the double-layer capacitance for the restricted primitive model: the effect of chemical association between unlike ions. PMID- 16035875 TI - Comment on "multireference configuration-interaction calculations for positronium halides" [J. Chem. Phys. 122, 054302 (2005)]. PMID- 16035877 TI - Construction of a natural partition of incomplete horseshoes. AB - We present a method for constructing a partition of an incomplete horseshoe in a Poincare map. The partition is based only on the unstable manifolds of the outermost fixed points and eventually their limits. Consequently, this partition becomes natural from the point of view of asymptotic scattering observations. The symbolic dynamics derived from this partition coincides with the one derived from the hierarchical structure of the singularities of the scattering functions. PMID- 16035878 TI - Fractional Fokker-Planck equation for fractal media. AB - We consider the fractional generalizations of equation that defines the medium mass. We prove that the fractional integrals can be used to describe the media with noninteger mass dimensions. Using fractional integrals, we derive the fractional generalization of the Chapman-Kolmogorov equation (Smolukhovski equation). In this paper fractional Fokker-Planck equation for fractal media is derived from the fractional Chapman-Kolmogorov equation. Using the Fourier transform, we get the Fokker-Planck-Zaslavsky equations that have fractional coordinate derivatives. The Fokker-Planck equation for the fractal media is an equation with fractional derivatives in the dual space. PMID- 16035879 TI - Spiking dynamics of interacting oscillatory neurons. AB - Spiking sequences emerging from dynamical interaction in a pair of oscillatory neurons are investigated theoretically and experimentally. The model comprises two unidirectionally coupled FitzHugh-Nagumo units with modified excitability (MFHN). The first (master) unit exhibits a periodic spike sequence with a certain frequency. The second (slave) unit is in its excitable mode and responds on the input signal with a complex (chaotic) spike trains. We analyze the dynamic mechanisms underlying different response behavior depending on interaction strength. Spiking phase maps describing the response dynamics are obtained. Complex phase locking and chaotic sequences are investigated. We show how the response spike trains can be effectively controlled by the interaction parameter and discuss the problem of neuronal information encoding. PMID- 16035880 TI - Impulsive synchronization of chaotic systems. AB - The issue of impulsive synchronization of a class of chaotic systems is investigated. Based on the impulsive theory and linear matrix inequality technique, some less conservative and easily verified criteria for impulsive synchronization of chaotic systems are derived. The proposed method is applied to the original Chua oscillators, and the corresponding synchronization conditions are obtained. Moreover, the boundary of the stable region is also estimated in terms of the equidistant impulse interval. The effectiveness of our method is shown by computer simulation. PMID- 16035881 TI - Stochastic multiresonance in the coupled relaxation oscillators. AB - We study the noise-dependent dynamics in a chain of four very stiff excitable oscillators of the FitzHugh-Nagumo type locally coupled by inhibitor diffusion. We could demonstrate frequency- and noise-selective signal acceptance which is based on several noise-supported stochastic attractors that arise owing to slow variable diffusion between identical excitable elements. The attractors have different average periods distinct from that of an isolated oscillator and various phase relations between the elements. We explain the correspondence between the noise-supported stochastic attractors and the observed resonance peaks in the curves for the linear response versus signal frequency. PMID- 16035882 TI - Using chaotic forcing to detect damage in a structure. AB - In this work we develop a numerical test for Holder continuity and apply it and another test for continuity to the difficult problem of detecting damage in structures. We subject a thin metal plate with incremental damage to the plate changes, its filtering properties, and therefore the phase space trajectories of the response chaotic excitation of various bandwidths. Damage to the plate changes its filtering properties and therefore the phase space of the response. Because the data are multivariate (the plate is instrumented with multiple sensors) we use a singular value decomposition of the set of the output time series to reduce the embedding dimension of the response time series. We use two geometric tests to compare an attractor reconstructed from data from an undamaged structure to that reconstructed from data from a damaged structure. These two tests translate to testing for both generalized and differentiable synchronization between responses. We show loss of synchronization of responses with damage to the structure. PMID- 16035883 TI - The phase-modulated logistic map. AB - We study the logistic mapping with the nonlinearity parameter varied through a delayed feedback mechanism. This history dependent modulation through a phaselike variable offers an enhanced possibility for stabilization of periodic dynamics. Study of the system as a function of nonlinearity and modulation parameters reveals new phenomena: In addition to period-doubling and tangent bifurcations, there can be bifurcations where the period increases by unity. These are extensions of crises that arise in nonlinear dynamical systems. Periodic orbits in this system can be systematized via the kneading theory, which in the present case extends the analysis of Metropolis, Stein, and Stein for unimodal maps. PMID- 16035884 TI - Meanders and reconnection-collision sequences in the standard nontwist map. AB - New global periodic orbit collision and separatrix reconnection scenarios exhibited by the standard nontwist map are described in detail, including exact methods for determining reconnection thresholds, methods that are implemented numerically. Results are compared to a parameter space breakup diagram for shearless invariant curves. The existence of meanders, invariant tori that are not graphs, is demonstrated numerically for both odd and even period reconnection for certain regions in parameter space. Implications for transport are discussed. PMID- 16035885 TI - Public-key encryption based on generalized synchronization of coupled map lattices. AB - Currently used public-key cryptosystems are based on difficulties in solving certain numeric theoretic problems, in which the way to predict the private key from the knowledge of the public key is computationally infeasible. Here we propose a method of constructing public-key cryptosystems by generalized synchronization of coupled map lattices, in which the difficulty in predicting the synchronous function is used as the trap-door function to deduce the private key from the public key. In specific, we implement this idea on the method of "Merkle's puzzles," and find that, incorporated with the chaotic dynamics, this traditional method is equipped with some new features and can be practical in certain situations. PMID- 16035886 TI - Lag and anticipating synchronization without time-delay coupling. AB - We describe a new method for achieving approximate lag and anticipating synchronization in unidirectionally coupled chaotic oscillators. The method uses a specific parameter mismatch between the drive and response that is a first order approximation to true time-delay coupling. As a result, an adjustable lag or anticipation effect can be achieved without the need for a variable delay line, making the method simpler and more economical to implement in many physical systems. We present a stability analysis, demonstrate the method numerically, and report experimental observation of the effect in radio-frequency electronic oscillators. In the circuit experiments, both lag and anticipation are controlled by tuning a single capacitor in the response oscillator. PMID- 16035887 TI - Efficient topological chaos embedded in the blinking vortex system. AB - We consider the particle mixing in the plane by two vortex points appearing one after the other, called the blinking vortex system. Mathematical and numerical studies of the system reveal that the chaotic particle mixing, i.e., the chaotic advection, is observed due to the homoclinic chaos, but the mixing region is restricted locally in the neighborhood of the vortex points. The present article shows that it is possible to realize a global and efficient chaotic advection in the blinking vortex system with the help of the Thurston-Nielsen theory, which classifies periodic orbits for homeomorphisms in the plane into three types: periodic, reducible, and pseudo-Anosov (pA). It is mathematically shown that periodic orbits of pA type generate a complicated dynamics, which is called topological chaos. We show that the combination of the local chaotic mixing due to the topological chaos and the dipole-like return orbits realize an efficient and global particle mixing in the blinking vortex system. PMID- 16035888 TI - Manifestation of scarring in a driven system with wave chaos. AB - We consider wave propagation in a model of a deep ocean acoustic wave guide with a periodic range dependence. It is assumed that the wave field is governed by the parabolic equation. Formally the mathematical model of the wave guide coincides with that of a quantum system with time-dependent Hamiltonian. From the analysis of Floquet modes of the wave guide it is shown that there exists a "scarring" effect similar to that observed in quantum systems. It turns out that the segments of an unstable periodic ray trajectory may be distinguished in the spatial distribution of the wave field intensity at a finite wavelength. Besides the scarring effect, it is found that the so-called "stable islands" in the phase space of ray dynamics reveal themselves in the coarse-grained Wigner functions of the Floquet modes. PMID- 16035889 TI - Breathers and thermal relaxation as a temporal process: a possible way to detect breathers in experimental situations. AB - Breather stability and longevity in thermally relaxing nonlinear arrays is investigated under the scrutiny of the analysis and tools employed for time series and state reconstruction of a dynamical system. We briefly review the methods used in the analysis and characterize a breather in terms of the results obtained with such methods. Our present work focuses on spontaneously appearing breathers in thermal Fermi-Pasta-Ulam arrays but we believe that the conclusions are general enough to describe many other related situations; the particular case described in detail is presented as another example of systems where three incommensurable frequencies dominate their chaotic dynamics (reminiscent of the Ruelle-Takens scenario for the appearance of chaotic behavior in nonlinear systems). This characterization may also be of great help for the discovery of breathers in experimental situations where the temporal evolution of a local variable (like the site energy) is the only available/measured data. PMID- 16035890 TI - Length scale competition in nonlinear Klein-Gordon models: a collective coordinate approach. AB - Working within the framework of nonlinear Klein-Gordon models as a paradigmatic example, we show that length scale competition, an instability of solitons subjected to perturbations of an specific length, can be understood by means of a collective coordinate approach in terms of soliton position and width. As a consequence, we provide a natural explanation of the phenomenon in much simpler terms than any previous treatment of the problem. Our technique allows us to study the existence of length scale competition in most soliton bearing nonlinear models and can be extended to coherent structures with more degrees of freedom. PMID- 16035891 TI - Restitution in mapping models with an arbitrary amount of memory. AB - Restitution, the characteristic shortening of action potential duration (APD) with increased heart rate, has been studied extensively because of its purported link to the onset of fibrillation. Restitution is often represented in the form of mapping models where APD is a function of previous diastolic intervals (DIs) and/or APDs, A(n+1)=F(D(n),A(n),D(n-1),A(n-1),...), where A(n+1) is the APD following a DI given by D(n). The number of variables previous to D(n) determines the degree of memory in the mapping model. Recent experiments have shown that mapping models should contain at least three variables (D(n),A(n),D(n-1)) to reproduce a restitution portrait (RP) that is qualitatively similar to that seen experimentally, where the RP shows three different types of restitution curves (RCs) [dynamic, S1-S2, and constant-basic cycle length (BCL)] simultaneously. However, an interpretation of the different RCs has only been presented in detail for mapping models of one and two variables. Here we present an analysis of the different RCs in the RP for mapping models with an arbitrary amount of memory. We determine the number of variables necessary to represent the different RCs in the RP. We also present a graphical visualization of these RCs. Our analysis reveals that the dynamic and S1-S2 RCs reside on two-dimensional surfaces, and therefore provide limited information for mapping models with more than two variables. However, constant-BCL restitution is a feature of the RP that depends on higher dimensions and can possibly be used to determine a lower bound on the dimensionality of cardiac dynamics. PMID- 16035892 TI - Effect of noise on defect chaos in a reaction-diffusion model. AB - The influence of noise on defect chaos due to breakup of spiral waves through Doppler and Eckhaus instabilities is investigated numerically with a modified Fitzhugh-Nagumo model. By numerical simulations we show that the noise can drastically enhance the creation and annihilation rates of topological defects. The noise-free probability distribution function for defects in this model is found not to fit with the previously reported squared-Poisson distribution. Under the influence of noise, the distributions are flattened, and can fit with the squared-Poisson or the modified-Poisson distribution. The defect lifetime and diffusive property of defects under the influence of noise are also checked in this model. PMID- 16035893 TI - Self-oscillations in chemoresponsive gels: a theoretical approach. AB - Some gels can swell or shrink in function of the concentration of the solutes contained in the solvent. We develop and discuss an algorithm based on Stefan Maxwell equations to study the dynamics of a sphere of chemoresponsive gel immersed in a medium kept far from equilibrium. We show that the coupling of the reaction with these volume changes creates some feedback that can lead to chemomechanical instabilities. As a result, a reaction that is not oscillatory in homogeneous conditions or in a nonresponsive gel can induce self-oscillations of volume and concentrations in a responsive gel. We also show that chemical oscillations related to long range activation can be strongly modified in such gels, even when the changes of volume are small. PMID- 16035894 TI - Synchronized firing of FitzHugh-Nagumo neurons by noise. AB - We investigate the influence of noise on synchronization between the spiking activities of neurons with external impulsive forces. We first analyze the dependence of the synchronized firing on the amplitude and the angular frequency of the impulsive force in the noise-free system. Three cases (regular spiking, traveling wave, and chaotic spiking) with low synchronized firing are chosen to study effects due to noise. In each case we find that small noise can be a promoter of synchronization phenomena in neural activities, by choosing an appropriate noise intensity acting on some of the neurons. PMID- 16035895 TI - Complete synchronization of the noise-perturbed Chua's circuits. AB - In this paper, complete synchronization between unidirectionally coupled Chua's circuits within stochastic perturbation is investigated. Sufficient conditions of complete synchronization between these noise-perturbed circuits are established by means of the so-called LaSalle-type invariance principle for stochastic differential equations. Specific examples and their numerical simulations are also provided to demonstrate the feasibility of these conditions. Furthermore, the results obtained for the coupled Chua's circuits are further generalized to the wide class of coupled systems within stochastic perturbation. PMID- 16035896 TI - Specific external forcing of spatiotemporal dynamics in reaction-diffusion systems. AB - Self-organization behavior and in particular pattern forming spatiotemporal dynamics play an important role in far from equilibrium chemical and biochemical systems. Specific external forcing and control of self-organizing processes might be of great benefit in various applications ranging from technical systems to modern biomedical research. We demonstrate that in a cellular chemotaxis system modeled by one-dimensional reaction-diffusion equations particular forms of spatiotemporal dynamics can be induced and stabilized by controlling spatially distributed influx patterns of a chemical species as a function of time. In our model study we show that a propagating wave with certain shape and velocity and static symmetrical and asymmetrical patterns can be forced and manipulated by numerically computing open-loop optimal influx controls. PMID- 16035897 TI - Q-S (lag or anticipated) synchronization backstepping scheme in a class of continuous-time hyperchaotic systems--a symbolic-numeric computation approach. AB - First, a Q-S (lag or anticipated) synchronization of continuous-time dynamical systems is defined. Second, based on a backstepping design with one controller, a systematic, concrete, and automatic scheme is developed to investigate the Q-S (lag or anticipated) synchronization between the drive system and response system with a strict-feedback form. Two identical hyperchaotic Tamasevicius-Namajunas Cenys(TNC) systems as well as the hyperchaotic TNC system and hyperchaotic Rossler system are chosen to illustrate the proposed scheme. Numerical simulations are used to verify the effectiveness of the proposed scheme. The scheme can also be extended to study Q-S (lag or anticipated) synchronization between other dynamical systems with strict-feedback forms. With the aid of symbolic-numeric computation, the scheme can be performed to yield automatically the scalar controller in computer. PMID- 16035898 TI - Neural mechanism for binaural pitch perception via ghost stochastic resonance. AB - We present a physiologically plausible binaural mechanism for the perception of the pitch of complex sounds via ghost stochastic resonance. In this scheme, two neurons are driven by noise and a different periodic signal each (with frequencies f(1)=kf(0) and f(2)=(k+1)f(0), where k>1), and their outputs (plus noise) are applied synaptically to a third neuron. Our numerical results, using the Morris-Lecar neuron model with chemical synapses explicitly considered, show that intermediate noise levels enhance the response of the third neuron at frequencies close to f(0), as in the cases previously described of ghost resonance. For the case of an inharmonic combination of inputs (f(1)=kf(0)+Deltaf and f(2)=(k+1)f(0)+Deltaf) noise is also seen to enhance the rates of most probable spiking for the third neuron at a frequency f(r)=f(0)+[Deltaf(k+12)]. In addition, we show that similar resonances can be observed as a function of the synaptic time constant. The suggested ghost-resonance-based stochastic mechanism can thus arise either at the peripheral level or at a higher level of neural processing in the perception of pitch. PMID- 16035899 TI - On the intrinsic time scales involved in synchronization: a data-driven approach. AB - We address the problem of detecting, from scalar observations, the time scales involved in synchronization of complex oscillators with several spectral components. Using a recent data-driven procedure for analyzing nonlinear and nonstationary signals [Huang, Proc. R. Soc. London A 454, 903 (1998)], we decompose a time series in distinct oscillation modes which may display a time varying spectrum. When applied to coupled oscillators with multiple time scales, we found that motions are captured in a finite number of phase-locked oscillations. Further, in the synchronized state distinct phenomena as phase slips, anti-phase or perfect phase locking can be simultaneously observed at specific time scales. This fully data-driven approach (without a priori choice of filters or basis functions) is tested on numerical examples and illustrated on electric intracranial signals recorded from an epileptic patient. Implications for the study of the build-up of synchronized states in nonstationary and noisy systems are pointed out. PMID- 16035900 TI - Perturbation parameters associated with nonlinear responses of the head at small amplitudes. AB - The head-neck system has multiple degrees of freedom in both its control and response characteristics, but is often modeled as a single joint mechanical system. In this study, we have attempted to quantify the perturbation parameters that would elicit nonlinear responses in a single degree-of-freedom neuromechanical system at small amplitudes and velocities of perturbation. Twelve healthy young adults seated on a linear sled randomly received anterior-posterior sinusoidal translations with +/-15 mm and +/-25 mm peak displacements at 0.81, 1.76, and 2.25 Hz. Head angular velocity and angular position data were examined using a nonlinear phase-plane analysis. Poincare sections of the phase plane were computed and Lyapunov exponents calculated to measure divergence (chaotic behavior) or convergence (stable behavior) of system dynamics. Variability of head angular position and velocity across the entire phase plot was compared to that of the Poincare sections to quantify spatial-temporal irregularity. Multiple equilibrium points and positive Lyapunov exponents revealed chaotic behavior at 0.81 Hz at both amplitudes whereas responses at 1.76 and 2.25 Hz exhibited periodic oscillations, clustered phase points, and negative Lyapunov exponents. However, intersubject variability increased at the lowest frequency and a few subjects presented chaotic behavior at all frequencies. An inverted pendulum with position and velocity threshold nonlinearity was adopted as a simplistic model of the head and neck. Simulations with the model resulted in features similar to those observed in the experimental data. Our principal finding was that increasing the perturbation amplitude had a stabilizing effect on the behavior across frequencies. Nonlinear behaviors observed at the lowest stimulus frequency might be attributed to fluctuations in control between the multiple sensory inputs. Although this study has not conclusively pointed toward any single mechanism as responsible for the responses observed, it has revealed clear directions for further investigation. To examine if changing the sensory modalities would elicit a significant change in the nonlinear behaviors observed here, further experiments that target a patient population with some sort of sensory deficit are warranted. PMID- 16035901 TI - Synchronization in power-law networks. AB - We consider realistic power-law graphs, for which the power-law holds only for a certain range of degrees. We show that synchronizability of such networks depends on the expected average and expected maximum degree. In particular, we find that networks with realistic power-law graphs are less synchronizable than classical random networks. Finally, we consider hybrid graphs, which consist of two parts: a global graph and a local graph. We show that hybrid networks, for which the number of global edges is proportional to the number of total edges, almost surely synchronize. PMID- 16035902 TI - Estimation of coupling between oscillators from short time series via phase dynamics modeling: limitations and application to EEG data. AB - We demonstrate in numerical experiments that estimators of strength and directionality of coupling between oscillators based on modeling of their phase dynamics [D. A. Smirnov and B. P. Bezruchko, Phys. Rev. E 68, 046209 (2003)] are widely applicable. Namely, although the expressions for the estimators and their confidence bands are derived for linear uncoupled oscillators under the influence of independent sources of Gaussian white noise, they turn out to allow reliable characterization of coupling from relatively short time series for different properties of noise, significant phase nonlinearity of the oscillators, and nonvanishing coupling between them. We apply the estimators to analyze a two channel human intracranial epileptic electroencephalogram (EEG) recording with the purpose of epileptic focus localization. PMID- 16035903 TI - Introduction: 100 years of Brownian motion. PMID- 16035904 TI - Brownian motion and diffusion: from stochastic processes to chaos and beyond. AB - One century after Einstein's work, Brownian motion still remains both a fundamental open issue and a continuous source of inspiration for many areas of natural sciences. We first present a discussion about stochastic and deterministic approaches proposed in the literature to model the Brownian motion and more general diffusive behaviors. Then, we focus on the problems concerning the determination of the microscopic nature of diffusion by means of data analysis. Finally, we discuss the general conditions required for the onset of large scale diffusive motion. PMID- 16035905 TI - From diffusion to anomalous diffusion: a century after Einstein's Brownian motion. AB - Einstein's explanation of Brownian motion provided one of the cornerstones which underlie the modern approaches to stochastic processes. His approach is based on a random walk picture and is valid for Markovian processes lacking long-term memory. The coarse-grained behavior of such processes is described by the diffusion equation. However, many natural processes do not possess the Markovian property and exhibit anomalous diffusion. We consider here the case of subdiffusive processes, which correspond to continuous-time random walks in which the waiting time for a step is given by a probability distribution with a diverging mean value. Such a process can be considered as a process subordinated to normal diffusion under operational time which depends on this pathological waiting-time distribution. We derive two different but equivalent forms of kinetic equations, which reduce to known fractional diffusion or Fokker-Planck equations for waiting-time distributions following a power law. For waiting time distributions which are not pure power laws one or the other form of the kinetic equation is advantageous, depending on whether the process slows down or accelerates in the course of time. PMID- 16035906 TI - The subtle nature of financial random walks. AB - We first review the most important "stylized facts" of financial time series, that turn out to be, to a large extent, universal. We then recall how the multifractal random walk of Bacry, Muzy, and Delour generalizes the standard model of financial price changes and accounts in an elegant way for many of their empirical properties. In a second part, we provide empirical evidence for a very subtle compensation mechanism that underlies the random nature of price changes. This compensation drives the market close to a critical point, that may explain the sensitivity of financial markets to small perturbations, and their propensity to enter bubbles and crashes. We argue that the resulting unpredictability of price changes is very far from the neoclassical view that markets are informationally efficient. PMID- 16035907 TI - Fundamental aspects of quantum Brownian motion. AB - With this work we elaborate on the physics of quantum noise in thermal equilibrium and in stationary nonequilibrium. Starting out from the celebrated quantum fluctuation-dissipation theorem we discuss some important consequences that must hold for open, dissipative quantum systems in thermal equilibrium. The issue of quantum dissipation is exemplified with the fundamental problem of a damped harmonic quantum oscillator. The role of quantum fluctuations is discussed in the context of both, the nonlinear generalized quantum Langevin equation and the path integral approach. We discuss the consequences of the time-reversal symmetry for an open dissipative quantum dynamics and, furthermore, point to a series of subtleties and possible pitfalls. The path integral methodology is applied to the decay of metastable states assisted by quantum Brownian noise. PMID- 16035909 TI - Non-Markovian stochastic processes: colored noise. AB - We survey classical non-Markovian processes driven by thermal equilibrium or nonequilibrium (nonthermal) colored noise. Examples of colored noise are presented. For processes driven by thermal equilibrium noise, the fluctuation dissipation relation holds. In consequence, the system has to be described by a generalized (integro-differential) Langevin equation with a restriction on the damping integral kernel: Its form depends on the correlation function of noise. For processes driven by nonequilibrium noise, there is no such a restriction: They are considered to be described by stochastic differential (Ito- or Langevin type) equations with an independent noise term. For the latter, we review methods of analysis of one-dimensional systems driven by Ornstein-Uhlenbeck noise. PMID- 16035908 TI - Quantum Brownian motion with large friction. AB - Quantum Brownian motion in the strong friction limit is studied based on the exact path integral formulation of dissipative systems. In this limit the time nonlocal reduced dynamics can be cast into an effective equation of motion, the quantum Smoluchowski equation. For strongly condensed phase environments it plays a similar role as master equations in the weak coupling range. Applications for chemical, mesoscopic, and soft matter systems are discussed and reveal the substantial role of quantum fluctuations. PMID- 16035910 TI - Self-diffusion in granular gases: Green-Kubo versus Chapman-Enskog. AB - We study the diffusion of tracers (self-diffusion) in a homogeneously cooling gas of dissipative particles, using the Green-Kubo relation and the Chapman-Enskog approach. The dissipative particle collisions are described by the coefficient of restitution epsilon which for realistic material properties depends on the impact velocity. First, we consider self-diffusion using a constant coefficient of restitution, epsilon=const, as frequently used to simplify the analysis. Second, self-diffusion is studied for a simplified (stepwise) dependence of epsilon on the impact velocity. Finally, diffusion is considered for gases of realistic viscoelastic particles. We find that for epsilon=const both methods lead to the same result for the self-diffusion coefficient. For the case of impact-velocity dependent coefficients of restitution, the Green-Kubo method is, however, either restrictive or too complicated for practical application, therefore we compute the diffusion coefficient using the Chapman-Enskog method. We conclude that in application to granular gases, the Chapman-Enskog approach is preferable for deriving kinetic coefficients. PMID- 16035911 TI - Random walks, diffusion limited aggregation in a wedge, and average conformal maps. AB - We investigate diffusion-limited aggregation (DLA) in a wedge geometry. Arneodo and collaborators have suggested that the ensemble average of DLA cluster density should be close to the noise-free selected Saffman-Taylor finger. We show that a different, but related, ensemble average, that of the conformal maps associated with random clusters, yields a nontrivial shape which is also not far from the Saffman-Taylor finger. However, we have previously demonstrated that the same average of DLA in a channel geometry is not the Saffman-Taylor finger. This casts doubt on the idea that the average of noisy diffusion-limited growth is governed by a simple transcription of noise-free results. PMID- 16035912 TI - Noise-assisted transport on symmetric periodic substrates. AB - The rectification of a massive Brownian particle moving on a periodic substrate can be achieved in the absence of spatial asymmetry, by having recourse to (at least) two periodic, zero-mean input signals. We determine the relevant drift current under diverse operation conditions, namely, additive and multiplicative couplings, adiabatic and fast oscillating drives, and propagating substrate modulations. Distinct rectification mechanisms result from the interplay of noise and commensuration of the input frequencies, mediated through the nonlinearity of the substrate. These mechanisms are then extended to characterize soliton transport along a directed multistable chain. As the side-wise soliton diffusion is ultimately responsible for the transverse diffusion of such chains, our approach provides a full account of the Brownian motion of both pointlike and linear objects on a periodic substrate. PMID- 16035913 TI - Performance characteristics of Brownian motors. AB - Brownian motors are nonequilibrium systems that rectify thermal fluctuations to achieve directed motion, using spatial or temporal asymmetry. We provide a tutorial introduction to this basic concept using the well-known example of a flashing ratchet, discussing the micro- to nanoscopic scale on which such motors can operate. Because of the crucial role of thermal noise, the characterization of the performance of Brownian motors must include their fluctuations, and we review suitable performance measures for motor coherency and efficiency. Specifically, we highlight that it is possible to determine the energy efficiency of Brownian motors by measuring their velocity fluctuations, without detailed knowledge of the motor function and its energy input. Finally, we exemplify these concepts using a model for an artificial single-molecule motor with internal degrees of freedom. PMID- 16035914 TI - Controlling the motion of interacting particles: homogeneous systems and binary mixtures. AB - We elaborate on recent results on the transport of interacting particles for both single-species and binary mixtures subject to an external driving on a ratchetlike asymmetric substrate. Moreover, we also briefly review motion control without any spatial asymmetric potential (i.e., no ratchet). Our results are obtained using an analytical approach based on a nonlinear Fokker-Planck equation as well as via numerical simulations. By increasing the particle density, the net dc ratchet current in our alternating (ac)-driven systems can either increase or decrease depending on the temperature, the drive amplitude, and the nature of the inter-particle interactions. This provides an effective control of particle motion by just changing the particle density. At low temperatures, attracting particles can condense at some potential minima, thus breaking the discrete translational symmetry of the substrate. Depending on the drive amplitude, an agglomeration or condensation results either in a drop to zero or in a saturation of the net particle velocity at densities above the condensation density-the latter case producing a very efficient rectification mechanism. For binary mixtures we find three ways of controlling the particle motion of one (passive) B species by means of another (active) A species: (i) Dragging the target particles B by driving the auxiliary particles A, (ii) rectifying the motion of the B particles on the asymmetric potential created by the A-B interactions, and (iii) dynamically modifying (pulsating) this potential by controlling the motion of the A particles. This allows to easily control the magnitude and direction of the velocity of the target particles by changing either the frequency, phase and/or amplitude of the applied ac drive(s). PMID- 16035915 TI - Moving backward noisily. AB - We discuss the fundamental physical differences and the mathematical interconnections of counterintuitive transport and response properties in Brownian motion far from equilibrium. After reviewing the ubiquity of such effects in physical and other systems, we illustrate the general properties on paradigmatic models for both individually and collectively acting Brownian particles. PMID- 16035916 TI - Colloids as model systems for problems in statistical physics. AB - Owing to their mesoscopic length scales, colloidal suspensions provide ideal model systems suitable for addressing many problems in the field of statistical physics. Exemplarily, we highlight the versatile nature of such systems by discussing experiments with stochastic resonance and a practical realization of a recently proposed ratchet cellular automaton. PMID- 16035917 TI - Stochastic resonance: theory and numerics. AB - We address the phenomenon of stochastic resonance in a noisy bistable system driven by a time-dependent periodic force (not necessarily sinusoidal) and in its two-state approximation. Even for driving forces with subthreshold amplitudes, the behavior of the system response might require a nonlinear description. We introduce analytical and numerical tools to analyze the power spectral amplification and the signal-to-noise ratio in a nonlinear regime. Our analysis shows the importance of the effects of the driving force on the system fluctuations in a nonlinear regime. These effects can be usefully exploited to achieve high quality output signals with gains larger than unity, which is impossible within a linear regime. PMID- 16035918 TI - Reaction rate theory: what it was, where is it today, and where is it going? AB - A brief history is presented, outlining the development of rate theory during the past century. Starting from Arrhenius [Z. Phys. Chem. 4, 226 (1889)], we follow especially the formulation of transition state theory by Wigner [Z. Phys. Chem. Abt. B 19, 203 (1932)] and Eyring [J. Chem. Phys. 3, 107 (1935)]. Transition state theory (TST) made it possible to obtain quick estimates for reaction rates for a broad variety of processes even during the days when sophisticated computers were not available. Arrhenius' suggestion that a transition state exists which is intermediate between reactants and products was central to the development of rate theory. Although Wigner gave an abstract definition of the transition state as a surface of minimal unidirectional flux, it took almost half of a century until the transition state was precisely defined by Pechukas [Dynamics of Molecular Collisions B, edited by W. H. Miller (Plenum, New York, 1976)], but even this only in the realm of classical mechanics. Eyring, considered by many to be the father of TST, never resolved the question as to the definition of the activation energy for which Arrhenius became famous. In 1978, Chandler [J. Chem. Phys. 68, 2959 (1978)] finally showed that especially when considering condensed phases, the activation energy is a free energy, it is the barrier height in the potential of mean force felt by the reacting system. Parallel to the development of rate theory in the chemistry community, Kramers published in 1940 [Physica (Amsterdam) 7, 284 (1940)] a seminal paper on the relation between Einstein's theory of Brownian motion [Einstein, Ann. Phys. 17, 549 (1905)] and rate theory. Kramers' paper provided a solution for the effect of friction on reaction rates but left us also with some challenges. He could not derive a uniform expression for the rate, valid for all values of the friction coefficient, known as the Kramers turnover problem. He also did not establish the connection between his approach and the TST developed by the chemistry community. For many years, Kramers' theory was considered as providing a dynamic correction to the thermodynamic TST. Both of these questions were resolved in the 1980s when Pollak [J. Chem. Phys. 85, 865 (1986)] showed that Kramers' expression in the moderate to strong friction regime could be derived from TST, provided that the bath, which is the source of the friction, is handled at the same level as the system which is observed. This then led to the Mel'nikov-Pollak-Grabert-Hanggi [Mel'nikov and Meshkov, J. Chem. Phys. 85, 1018 (1986); Pollak, Grabert, and Hanggi, ibid. 91, 4073 (1989)] solution of the turnover problem posed by Kramers. Although classical rate theory reached a high level of maturity, its quantum analog leaves the theorist with serious challenges to this very day. As noted by Wigner [Trans. Faraday Soc. 34, 29 (1938)], TST is an inherently classical theory. A definite quantum TST has not been formulated to date although some very useful approximate quantum rate theories have been invented. The successes and challenges facing quantum rate theory are outlined. An open problem which is being investigated intensively is rate theory away from equilibrium. TST is no longer valid and cannot even serve as a conceptual guide for understanding the critical factors which determine rates away from equilibrium. The nonequilibrium quantum theory is even less well developed than the classical, and suffers from the fact that even today, we do not know how to solve the real time quantum dynamics for systems with "many" degrees of freedom. PMID- 16035919 TI - Noise induced complexity: from subthreshold oscillations to spiking in coupled excitable systems. AB - We study the stochastic dynamics of an ensemble of N globally coupled excitable elements. Each element is modeled by a FitzHugh-Nagumo oscillator and is disturbed by independent Gaussian noise. In simulations of the Langevin dynamics we characterize the collective behavior of the ensemble in terms of its mean field and show that with the increase of noise the mean field displays a transition from a steady equilibrium to global oscillations and then, for sufficiently large noise, back to another equilibrium. In the course of this transition diverse regimes of collective dynamics ranging from periodic subthreshold oscillations to large-amplitude oscillations and chaos are observed. In order to understand the details and mechanisms of these noise-induced dynamics we consider the thermodynamic limit N-->infinity of the ensemble, and derive the cumulant expansion describing temporal evolution of the mean field fluctuations. In Gaussian approximation this allows us to perform the bifurcation analysis; its results are in good qualitative agreement with dynamical scenarios observed in the stochastic simulations of large ensembles. PMID- 16035920 TI - Multiple locus linkage analysis of genomewide expression in yeast. AB - With the ability to measure thousands of related phenotypes from a single biological sample, it is now feasible to genetically dissect systems-level biological phenomena. The genetics of transcriptional regulation and protein abundance are likely to be complex, meaning that genetic variation at multiple loci will influence these phenotypes. Several recent studies have investigated the role of genetic variation in transcription by applying traditional linkage analysis methods to genomewide expression data, where each gene expression level was treated as a quantitative trait and analyzed separately from one another. Here, we develop a new, computationally efficient method for simultaneously mapping multiple gene expression quantitative trait loci that directly uses all of the available data. Information shared across gene expression traits is captured in a way that makes minimal assumptions about the statistical properties of the data. The method produces easy-to-interpret measures of statistical significance for both individual loci and the overall joint significance of multiple loci selected for a given expression trait. We apply the new method to a cross between two strains of the budding yeast Saccharomyces cerevisiae, and estimate that at least 37% of all gene expression traits show two simultaneous linkages, where we have allowed for epistatic interactions. Pairs of jointly linking quantitative trait loci are identified with high confidence for 170 gene expression traits, where it is expected that both loci are true positives for at least 153 traits. In addition, we are able to show that epistatic interactions contribute to gene expression variation for at least 14% of all traits. We compare the proposed approach to an exhaustive two-dimensional scan over all pairs of loci. Surprisingly, we demonstrate that an exhaustive two-dimensional scan is less powerful than the sequential search used here. In addition, we show that a two-dimensional scan does not truly allow one to test for simultaneous linkage, and the statistical significance measured from this existing method cannot be interpreted among many traits. PMID- 16035921 TI - De novo formation of left-right asymmetry by posterior tilt of nodal cilia. AB - In the developing mouse embryo, leftward fluid flow on the ventral side of the node determines left-right (L-R) asymmetry. However, the mechanism by which the rotational movement of node cilia can generate a unidirectional flow remains hypothetical. Here we have addressed this question by motion and morphological analyses of the node cilia and by fluid dynamic model experiments. We found that the cilia stand, not perpendicular to the node surface, but tilted posteriorly. We further confirmed that such posterior tilt can produce leftward flow in model experiments. These results strongly suggest that L-R asymmetry is not the descendant of pre-existing L-R asymmetry within each cell but is generated de novo by combining three sources of spatial information: antero-posterior and dorso-ventral axes, and the chirality of ciliary movement. PMID- 16035922 TI - 'Lost in translation'? Developing assessment criteria that value rural practice. AB - INTRODUCTION: Rural workforce preparation is often discussed in terms of specific interventions such as rural placements. More technical discussions of education matters seem to belong in the realm of education experts. However, this issues article argues that a focus on quality assessment techniques is important to the rural health agenda. Making connections between the medical education literature and the broader education literature, it explores elements of a qualitative decision-making model as an alternative to narrow competency-based and norm referenced approaches. In the process it explores assessment techniques that may help educators better translate their intentions to value rural practice into the learning of students. BACKGROUND: Research suggests that, in Australia at least, many university educators have different and conflicting understanding of assessment criteria. At the same time, the literature on the development of assessment criteria is relatively small in a context in which the medical education literature takes a quantitative, reliability-driven approach. This has important implications for how we ensure that rural practice is given enough emphasis at the level of education that most strongly drives student learning assessment. METHODS: This article explores such matters by examining the steps needed to develop assessment criteria in undergraduate medical education courses. It draws on key writings from the past, as well as current debates, in the medical education and broader education literature. It focuses on the detail of assessment techniques to show how the intention to value rural practice can be 'lost in translation' with narrow norm-referenced and competency-based assessment models. CONCLUSIONS: Rural health has a stake in technical debates about education in health sciences courses. Like other knowledge and skills, the knowledge and skills important to rural practice cannot be valued at the coalface of student learning if our assessment techniques subvert intentions. Developing the quality of assessment techniques involves scrutiny of not only the medical education literature, but also the broader education literature, including writings about working models of criteria-and-standards-based assessment. This scrutiny suggests assessment techniques are not equal in terms of how well they translate intentions. More than that, it suggests the value to rural health education of shifting from narrow norm-referenced models to best practice in criteria-and-standards-based assessment. PMID- 16035925 TI - Telethinking. Interview with David K. Ahern. PMID- 16035927 TI - Globalization, health, and the role of telemedicine. PMID- 16035928 TI - Telemedicine evaluation. AB - This paper examines the context and status of evaluation research in telemedicine, and it proposes a two-pronged strategy for addressing the critical policy and programmatic concerns in this field. It explains the evolution of evaluation research in the United States, and it describes a comprehensive typology and requirements for valid evaluation. Major impediments for definitive evaluation are discussed, together with a summary of major trends in empirical studies. Two concurrent strategies are proposed for producing definitive findings and for assessing the available empirical evidence. These consist of large-scale experimental studies and theoretical and empirical triangulation for assessing the available empirical evidence. PMID- 16035929 TI - The evaluation of telemedicine and health services research. PMID- 16035930 TI - Clinical outcomes associated with telemedicine/telehealth. AB - This paper is a comprehensive review and synthesis of the literature concerning clinical outcomes associated with various telemedicine applications. It starts out with a brief description of the findings reported by similar literature reviews already published. Subsequently, it proposes a conceptual model for assessing clinical outcomes based on Donabedian's formulation of the Medical Care Process. Accordingly, research findings are reported in terms of the relevant components of the medical care process, namely, diagnosis, clinical management, and clinical outcomes. Specific findings are organized according to the designated clinical and diagnostic application. This is followed by a general report of studies dealing with patient satisfaction. PMID- 16035931 TI - Research findings and strategies for assessing telemedicine costs. AB - With varying degrees of enthusiasm, researchers and decision-makers support the use of telemedicine. Forms of telemedicine are appearing in health-care delivery, and are often integral to transforming health-care information technology. Despite this, the appropriate role of telemedicine in the delivery process remains ambiguous, at least partly because of its uncertain impact on costs. Cost savings and benefits are often suggested by the logic of its impact on health care and by the promise of technology, but definitive information on the costs and benefits remain elusive. The objectives of this paper are to review the state of telemedicine cost research, to examine major issues affecting the yield from this research, and finally to recommend strategies for improving future research. As this paper demonstrates, the productivity of telemedicine cost studies suffers from an under-utilization of appropriate program evaluation and economic methods. This review of telemedicine cost literature will appraise telemedicine cost studies and their findings within a broad analytic framework. Telemedicine cost studies will be assessed on their methods of statistical inference, use of critical economic concepts, and contextual definition for the determination of costs and benefits. PMID- 16035932 TI - Evolving telemedicine/ehealth technology. AB - This paper describes emerging technologies to support a rapidly changing and expanding scope of telemedicine/telehealth applications. Of primary interest here are wireless systems, emerging broadband, nanotechnology, intelligent agent applications, and grid computing. More specifically, the paper describes the changes underway in wireless designs aimed at enhancing security; some of the current work involving the development of nanotechnology applications and research into the use of intelligent agents/artificial intelligence technology to establish what are termed "Knowbots"; and a sampling of the use of Web services, such as grid computing capabilities, to support medical applications. In addition, the expansion of these technologies and the need for cost containment to sustain future health care for an increasingly mobile and aging population is discussed. PMID- 16035933 TI - Toward technical interoperability in telemedicine. AB - For telemedicine to realize the vision of anywhere, anytime access to care, the question of how to create a fully interoperable technical infrastructure must be addressed. After briefly discussing how "technical interoperability" compares with other types of interoperability being addressed in the telemedicine community today, this paper describes reasons for pursuing technical interoperability, presents a proposed framework for realizing technical interoperability, identifies key issues that will need to be addressed if technical interoperability is to be achieved, and suggests a course of action that the telemedicine community might follow to accomplish this goal. PMID- 16035934 TI - Semantic interoperability in telemedicine through ontology-driven services. PMID- 16035938 TI - Recombinant Sendai virus as a novel vaccine candidate for respiratory syncytial virus. AB - Respiratory syncytial virus (RSV) is among the most important and serious pediatric respiratory diseases, and yet after more than four decades of research an effective vaccine is still unavailable. This review examines the role of the immune response in reducing disease severity; considers the history of RSV vaccine development; and advocates the potential utility of Sendai virus (a murine paramyxovirus) as a xenogenic vaccine vector for the delivery of RSV antigens. The immunogenicity and protective efficacy of RSV-recombinant Sendai virus vectors constructed using reverse genetics is examined. RSV-recombinant Sendai virus is easy to grow (i.e., achieves extremely high titers in eggs), is easy to administer (intranasal drops), and elicits both B- and T-cell responses leading to protection from RSV challenge in a small-animal model. Unmodified Sendai virus is currently being studied in clinical trials as a vaccine for its closely related human cognate (human parainfluenza virus type 1). Sendai virus may prove an enormously valuable vaccine platform, permitting the delivery of recombinants targeting important pediatric respiratory pathogens, RSV chief among them. PMID- 16035939 TI - Herpesviruses and the innate immune response. AB - Herpesvirus infection leads to the rapid induction of an innate immune response. A central aspect of this host response is the production and secretion of type I interferon. The current model of virus-mediated interferon production includes three stages: sensitization, induction, and amplification. A key mediator of all three stages is the cellular transcription factor interferon regulatory factor 3 (IRF3). Although the precise details of IRF3 activation and interferon production in response to herpesvirus infection are still being elucidated, viral proteins that block components of the interferon pathway, particularly IRF3, have been identified and characterized. In vivo studies have shown that in addition to type I interferon, interleukin-15 (IL-15) and natural killer (NK) cells also play an important role in mediating resistance to herpesvirus infection. Recent investigations have demonstrated a strong association between IRF3, interferon, IL-15, and NK cells. This review will focus on herpesvirus-mediated induction of innate immunity, the central role of the type I interferon response and mechanisms used by herpesviruses to block host antiviral immunity. PMID- 16035940 TI - Adenoviral vectors for mucosal vaccination against infectious diseases. AB - Adenoviral vector has been extensively studied as a vaccine platform because of its ability to induce potent cellular and humoral immunity. One main advantage of adenoviral vectors is their natural tropism for mucosal surfaces, which makes them ideal for the purpose of mucosal vaccination against pathogens that preferentially initiate infection at the mucosal site. The current understanding of mucosal immunity suggests that mucosal vaccination is far superior to parenteral vaccination in protecting mucosal surfaces. Mucosal vaccination is particularly relevant to those infections for which parenteral immunization strategies have failed to confer protection. This review examines the use of adenoviral vector at mucosal sites for infectious disease against which the current vaccination strategies have been unsuccessful in eliciting protection. Data from animal models have suggested that adenoviral vectors are effective in protecting against infections caused by HIV, herpes simplex virus and Mycobacterium tuberculosis. We believe that these encouraging results will lead to further evaluation in clinical trials in the near future. PMID- 16035941 TI - Cell-mediated immune response to human T-lymphotropic virus type I. AB - Human T-lymphotropic virus type I (HTLV-I) is a retrovirus that causes persistent infection in many populations in tropical and subtropical regions. HTLV-I chronically activates the cell-mediated arm of the host adaptive immune response. There has been much debate about the role of the immune response in determining the outcome of HTLV-I infection: most seropositive individuals remain lifelong asymptomatic carriers of the virus, whereas a small proportion-usually those with higher equilibrium proviral loads-develop an inflammatory disease of the central nervous system known as HAM/TSP. Here we discuss the cell-mediated immune response to HTLV-I infection. We summarize recent data on the HTLV-I-specific CD4(+) cell response and explore its potential role in HAM/TSP pathogenesis. We also explore the controversy surrounding the role of the CD8(+) cell response in controlling HTLV-I infection and/or contributing to HAM/TSP disease, highlighting recent studies of T cell gene expression profiles and a newly developed assay of CD8(+) cell functional efficiency. Finally, we introduce a possible role for cellular innate immune effectors in HTLV-I infection. PMID- 16035942 TI - Public fear of vaccination: separating fact from fiction. AB - During the last two centuries, the world has seen a substantial increase in the number and availability of vaccines for the prevention of infectious disease. Smallpox vaccine remains the most celebrated vaccine-related achievement in human history, but worldwide reductions in many other diseases including measles, mumps, rubella, polio, diphtheria, and whooping cough (Bordetella pertussis) also illustrate the power of vaccination in controlling outbreaks of contagious diseases. Ironically, as advances in vaccination successfully limit disease outbreaks, the impact that these infectious agents once had on society becomes marginalized. Public confidence in vaccination may erode because of real or perceived risks associated with immunization, and this in turn may lead to lower vaccination coverage and loss of herd immunity. Here, we will discuss some of the elements associated with public perceptions and fear of vaccination and place these into the context of how deadly several vaccine-preventable childhood diseases can be if vaccination coverage is insufficient. PMID- 16035943 TI - Live attenuated measles vaccine as a potential multivalent pediatric vaccination vector. AB - Live attenuated RNA viruses make highly efficient vaccines. Among them is the live attenuated measles virus (MV) vaccine that has been given to a very large number of children and has been shown to be highly efficacious and safe. MV vaccine induces a life-long immunity after a single injection or two low-dose injections. It is easily produced on a large scale in most countries and can be distributed at low cost. Reversion to pathogenicity has never been observed with this vaccine. For all of these characteristics, developing of MV vaccine vector as a multivalent vaccine to immunize children against both measles and other infectious agents such as human immunodeficiency virus (HIV), flaviviruses, or malaria might be very promising for worldwide use. As MV vaccine is inexpensive to produce, the generation of recombinant vaccines may remain affordable and attractive for the developing word. In this article, we describe the development of MV vector and present some recent data showing the capacity of recombinant MV vaccine to express various proteins from HIV and West Nile virus. In addition, the ability of recombinant MV to induce specific immune responses against these different pathogens are presented and discussed. PMID- 16035944 TI - Vaccine design for severe acute respiratory syndrome coronavirus. AB - Severe acute respiratory syndrome (SARS) is an emerging infectious disease caused by a new coronavirus (SARS-CoV). Recent studies suggest that SARS-CoV is zoonotic and may have a broad host range besides humans. Although the global outbreak of SARS has been contained, there are serious concerns over its re-emergence and bioterrorism potential. As a part of preparedness, development of a safe and effective vaccine is one of the highest priorities in fighting SARS. A number of candidate vaccines, using a variety of approaches, are under development. The first vaccine tested in clinical trial is made from the inactivated form of SARS CoV. Several live attenuated, genetically engineered or vector vaccines encoding the SARS-CoV spike (S) protein have been in pre-clinical studies. These vaccine candidates are effective in terms of eliciting protective immunity in the vaccinated animals. However, caution should be taken with the safety of whole virus or full-length S protein-based immunogens in humans because they may induce harmful immune or inflammatory responses. We propose to use the receptor-binding domain (RBD) of SARS-CoV S protein (residues 318--510) for developing a safe and effective subunit SARS vaccine, as it is not only a functional domain that mediates virus-receptor binding but also a major neutralization determinant of SARSCoV. It has been demonstrated that the RBD of SARS-CoV S protein contains multiple conformational epitopes capable of inducing highly potent neutralizing antibody responses and protective immunity. PMID- 16035945 TI - Correlation between type of adaptive immune response against porcine circovirus type 2 and level of virus replication. AB - Porcine circovirus 2 (PCV2) replication is characterized by high variation among infected pigs. This study investigated the role of immunologic responses in causing this variation. Twelve gnotobiotic pigs were inoculated with PCV2. Four of these pigs were treated with cyclosporin A (CysA) to monitor the effect of the adaptive immunity on the development of the PCV2 infection. Through lymph node biopsies at 10, 15, and 21 days postinoculation (DPI), PCV2 replication in lymphoid tissues was monitored. The production of total PCV2-specific and PCV2 neutralizing antibodies was followed, together with interferon-gamma (IFN-gamma) mRNA expression levels in peripheral blood monocytes as a marker for cellular immunity. In general, the CysA-treated pigs showed the highest PCV2 titers, indicating that the adaptive immunity is necessary to restrain PCV2 replication. Three different PCV2 replication patterns were observed in non-CysA-treated pigs. Pattern 1: In two pigs, PCV2 was not detected. They had the highest neutralizing antibody titers, appearing from 15 DPI. In these pigs a good cellular response was indicated by a peak in IFN-gamma mRNA at 15 DPI. Pattern 2: Five pigs contained low to moderate PCV2 titers at 15 DPI, remaining constant or decreasing towards 21 DPI. Lower neutralizing antibody titers were observed and no rise in IFN-gamma was detected. Pattern 3: In one pig, a low PCV2 titer at 15 DPI dramatically increased toward 21 DPI. Although an antibody response against PCV2 was mounted, no PCV2-neutralizing antibodies were detected. This pig also showed no rise in IFN-gamma. The study findings indicate that variation in the onset of the adaptive immunity may account for variation in PCV2 replication among pigs. Absence of PCV2-neutralizing antibodies may be an important factor in the development of an increased virus replication. PMID- 16035946 TI - N-linked glycosylation in C2 region of HIV-1 envelope reduces sensitivity to neutralizing antibodies. AB - N-linked glycosylation at specific sites on human immunodeficiency virus (HIV)--1 gp120 envelope glycoprotein is believed to act as a glycan shield to protect the viral neutralizing epitopes. Various glycosylation sites have been shown to affect the sensitivity to antibody-mediated neutralization. These include sites on V1V2, C2, base of V3, V5 and C5. Among these, the sites around the base of V3 loop have been most consistently found to associate with neutralization sensitivity in subtype B viruses. In contrast, we found that N-linked glycosylation sites at the junction of V2--C2 and in the middle of C2 were responsible for the neutralization resistance in CRF01_A/E, whereas sites at the base of V3 loop and in V1 and V5 did not affect the neutralization phenotype. PMID- 16035947 TI - Interferon-gamma-induced activation of nitric oxide-mediated antiviral activity of macrophages caused by a recombinant coxsackievirus B3. AB - Cardiovascular disease is one of the major causes of human death and has been linked to many different risks including viral infections. Coxsackievirus B3 (CVB3) is one of the most important pathogens responsible for virus-induced myocarditis. Cytokines are normally involved in the control of CVB3 replication and pathogenesis. Among them, interferon-gamma (IFN-gamma) in particular is highly protective against CVB3. A novel strategy to circumvent virus-caused heart disease is based on the development of cytokine-expressing recombinant virus vectors. Using in vitro co-culture experiments, the release of IFN-gamma by the recombinant virus variant CVB3/IFN-gamma activates the expression of the inducible nitric oxide synthase (iNOS) in CVB3 non-susceptible murine macrophages and the release of nitric oxide (NO), which reduce coxsackieviral replication directly. In addition, the expression of IFN-gamma by CVB3/IFN-gamma contributes to protect mice from lethal infections by iNOS induction in murine peritoneal macrophages, viral load reduction, and pancreatic tissue protection. PMID- 16035948 TI - Virus-like particle vaccine conferred complete protection against a lethal influenza virus challenge. AB - We have previously demonstrated the formation and release of influenza virus-like particles (VLPs) from the surface of Sf9 cells infected with either a quadruple baculovirus recombinant that simultaneously expresses the influenza structural proteins hemagglutinin (HA), neuraminidase (NA), matrix 1 (M1), and matrix 2 (M2), or a combination of single recombinants that include the M1 protein. In this work, we present data on the immunogenicity and protective efficacy afforded by VLPs (formed by M1 and HA) after immunization of mice. VLP vaccine ( approximately 1 microg HA) were formulated with or without IL-12 as adjuvant and administered twice, at 2-week intervals, by either intranasal instillation or intramuscular injection. All VLP-vaccinated and influenza-immunized control mice demonstrated high antibody titers to the HA protein; however, intranasal instillation of VLPs elicited antibody titers that were higher than those induced by either intramuscular inoculation of VLPs or intranasal inoculation with two sub-lethal doses of the challenge influenza virus (control group). Antibody responses were enhanced when VLP vaccine was formulated with IL12 as adjuvant. All mice were challenged with 5 LD50 of a mouse-adapted influenza A/Hong Kong/68 (H3N2) virus. Intramuscular administration of VLP vaccine formulated with or without IL-12 afforded 100% protection against a lethal influenza virus challenge. Similarly, intranasal instillation of VLP vaccine alone protected 100% of the mice, whereas VLP formulated with IL-12 protected 90% of the vaccinated mice. Not only do these results suggest a novel approach to the development of VLP vaccines for diverse influenza virus strains, but also the creation of multivalent vaccines by decoration of the surface of the VLPs with antigens from other pathogens. PMID- 16035949 TI - Cross-reactive protection against influenza A virus by a topically applied DNA vaccine encoding M gene with adjuvant. AB - The skin is rich with immunocompetent cells and therefore immunization through the skin is an attractive alternative to the invasive vaccination methods currently used. In this study the backs of mice were gently shaved, hydrated, and painted with a DNA vaccine encoding influenza M protein with adjuvant. The immunized mice were then challenged with two mouse-adapted strains of the influenza virus A: A/PR/8/34 (H1N1) and A/Udorn/72 (H3N2). This adjuvanated and topically applied DNA vaccine efficiently induced cytotoxic as well as humoral immune response and provide cross-reactive protection against several strains of influenza A virus. For better protection against virus infection, it will be necessary to select and combine the DNA vaccine with an appropriate adjuvant. PMID- 16035950 TI - Porcine reproductive and respiratory syndrome (PRRS) virus-specific interferon gamma(+) T-cell responses after PRRS virus infection or vaccination with an inactivated PRRS vaccine. AB - Although field studies have found porcine reproductive and respiratory syndrome (PRRSV) inactivated vaccines to be beneficial in reducing losses linked to PRRSV infection, immune mechanisms induced by these vaccines need better understanding. In the study reported here, we examined the interferon-gamma(+) (IFNgamma(+)) PRRS-specific T cell responses induced after infection and vaccination with an inactivated PRRS vaccine. Autologous monocyte-derived dendritic cells loaded with the PRRSV P120 strain were used to re-stimulate ex vivo T cells that had been primed in vivo by either the virus or the vaccine, or both. Virus-specific IFNgamma(+) T cells were quantified by using a porcine IFNgamma- ELISpot assay. A specific but low live virus-induced response was observed between days 35 and 70 for most of the pigs tested, while a significant inactivated vaccine-induced PRRSV-specific IFNgamma(+) T-cell response was measured soon after vaccination. Moreover, we observed that vaccination of pre-challenged pigs clearly favoured the PRRSV-specific cell-mediated immunity primed by the live virus. To characterize further the nature of the PRRSV-specific T cells, the different T cell subsets involved in PRRSV immunity were analyzed by flow cytometry. We showed that the inactivated vaccine was able to prime both CD4(+)CD8(int+) and CD8(high) virus-specific T cells and that CD4(+)CD8(int+) were preferentially recalled by the live virus. PMID- 16035951 TI - Human cytomegalovirus-specific CD4(+) T-cell clones recognize cross-reactive peptides from the immediate early 1 protein. AB - Human cytomegalovirus (HCMV) is a beta-herpes virus that persists in a latent state in immunocompetent individuals. Both CD4(+) and CD8(+) T lymphocytes have been reported to be present at a high frequency in HCMV-seropositive individuals and are involved in the control of infection. How such frequencies are maintained is not completely understood. We have observed that the canonical HLA-DR8 epitope of the immediate early 1 protein (IE1) contained in the IE1 (156--175) sequence shares homologies with an IE1 sequence contained in part in the previously reported HLA-DR3 epitope, IE1 (91-110). We thus wondered whether such homology in a single protein would translate into recognition of the IE1 homolog sequence by HLA-DR8-restricted CD4(+) cells in addition to the canonical epitope. We found that established HLA-DR8-restricted T cell clones are also able to cross recognize the IE1 (91--110) peptide, as well as a shorter 14-mer, IE1 (91--104). Moreover, the homolog peptide IE1 (91-110) was able to generate, from a seropositive blood donor, new IE1-specific, HLA-DR8-restricted CD4(+) T cell clones that were also cross-reactive. Those findings may provide clues to the formation and regulation of the T-cell repertoire and memory. PMID- 16035952 TI - Interleukin (IL)-12 receptor beta1 or IL-12 receptor beta 2 deficiency in mice indicates that IL-12 and IL-23 are not essential for host recovery from viral encephalitis. AB - Vesicular stomatitis virus (VSV), a negative-sense, single-stranded RNA rhabdovirus, causes acute viral encephalitis when administered intranasally to mice. Interleukin-12 (IL-12) is a key pro-inflammatory cytokine that is produced largely by the antigen presenting cells (APC) and that bridges the innate and acquired immune responses. IL-12 is efficacious in enhancing recovery from VSV infection of the murine central nervous system. This effect is mediated by nitric oxide (NO) produced by the neuronal isoform of nitric oxide synthase (NOS-1), and is independent of the pro-inflammatory cytokines interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha). These data implied a link between IL-12 and NOS-1. Here we investigate the role of the IL-12R during VSV pathogenesis, using IL-12R beta2 and IL-12R beta1-deficient mice. We showed that a deficiency in either IL-12R beta2 or IL-12R beta1 had no effect on the outcome of VSV infection of the CNS or on the clearance of VSV from the CNS. Furthermore, these data indicate that IL-23 is not acting redundantly in the absence of IL-12 during VSV-induced encephalitis. PMID- 16035955 TI - HIF1-alpha overexpression indicates a good prognosis in early stage squamous cell carcinomas of the oral floor. AB - BACKGROUND: Hypoxia-inducible factor 1 (HIF-1) is a transcription factor, which plays a central role in biologic processes under hypoxic conditions, especially concerning tumour angiogenesis. HIF-1alpha is the relevant, oxygen-dependent subunit and its overexpression has been associated with a poor prognosis in a variety of malignant tumours. Therefore, HIF-1alpha expression in early stage oral carcinomas was evaluated in relation to established clinico-pathological features in order to determine its value as a prognostic marker. METHODS: 85 patients with histologically proven surgically treated T1/2 squamous cell carcinoma (SCC) of the oral floor were eligible for the study. Tumor specimens were investigated by means of tissue micro arrays (TMAs) and immunohistochemistry for the expression of HIF-1. Correlations between clinical features and the expression of HIF-1 were evaluated by Kaplan-Meier curves, log-rank tests and multivariate Cox regression analysis. RESULTS: HIF-1alpha was frequently overexpressed in a probably non-hypoxia related fashion. The expression of HIF 1alpha was related with a significantly improved 5-year survival rate (p < 0.01) and a significantly increased disease free period (p = 0.01) independent from nodal status and tumour size. In primary node negative T1/T2 SCC of the oral floor, absence of HIF-1alpha expression specified a subgroup of high-risk patients (p < 0.05). CONCLUSION: HIF-1alpha overexpression is an indicator of favourable prognosis in T1 and T2 SCC of the oral floor. Node negative patients lacking HIF-1alpha expression may therefore be considered for adjuvant radiotherapy. PMID- 16035956 TI - The effects of celecoxib augmentation on cytokine levels in schizophrenia. AB - Celecoxib augmentation therapy has been reported to enhance the rate of clinical response for patients with schizophrenia. This may be due in part to an effect of celecoxib in the immune dysfunction associated with schizophrenia. Given concerns about the safety of COX-2 inhibitors, studies investigating cytokine levels in medicated patients with schizophrenia are of public health importance. Twenty eight schizophrenia subjects stabilized on olanzapine or risperidone were randomized to receive 8 wk of celecoxib (400 mg/d) or placebo. Serum soluble IL-2 receptor (sIL-2r) and in-vitro PHA-stimulated whole-blood cytokine production levels were measured at baseline, 1 wk, and 8 wk. Celecoxib augmentation did not alter any of the cytokine parameters measured for the overall study group. However, 1 wk of celecoxib augmentation increased TNF-alpha and IL-2 production levels in olanzapine-treated subjects. These elevations did not persist by week 8. Overall, celecoxib does not significantly modify cytokine levels in medicated schizophrenia subjects. PMID- 16035957 TI - A video method for the evaluation of antidepressant clinical and behavioural actions. AB - Measuring efficacy and behavioural actions of new antidepressants (ADs) is greatly enhanced by having videotaped records of assessment interviews. This study describes a revised version of the Video Interview Behaviour Evaluation Scales (VIBES), shortened to make it more applicable to clinical trials. The method focuses on physically expressive, motor and social behavioural aspects of the depressive disorder. The Brief version permits juxtaposing of baseline and outcome interviews during the same viewing session thereby reducing the role of memory in the rater's observations. The method provides measures of behavioural components and four new severity dimensions, Social withdrawal-motor retardation, Anxiety-agitation, Hostility, Depressive mood-cognitive impairment. Viewing a patient's series of baseline, during treatment, and outcome interviews can be conducted in about 1 h. This study compared the VIBES with the Hamilton Depression Rating Scale (HAMD) in (1) assessing the efficacy of the selectively targeted ADs, desipramine and paroxetine, and placebo in depressed in-patients and (2) determining onset and nature of the drugs' early behavioural actions. The findings showed (1) components of the method to be reliable; (2) the VIBES to be more sensitive than the HAMD in measuring efficacy; (3) the methods to be equally sensitive in detecting early clinical actions of the two drugs; (4) the VIBES more informative in identifying discrete behavioural aspects of the disorder that are impacted by the drugs; and (5) that in differentiating the drugs' behavioural actions, desipramine was indicated to initially 'stimulate', i.e. effect motor activity and depressed mood, and paroxetine to reduce global severity and anxiety. The study shows the VIBES to be capable of uncovering behavioural mechanisms underlying AD's capacity to resolve depressive disorder. PMID- 16035954 TI - Mouse behavioural analysis in systems biology. AB - Molecular techniques allowing in vivo modulation of gene expression have provided unique opportunities and challenges for behavioural studies aimed at understanding the function of particular genes or biological systems under physiological or pathological conditions. Although various animal models are available, the laboratory mouse (Mus musculus) has unique features and is therefore a preferred animal model. The mouse shares a remarkable genetic resemblance and aspects of behaviour with humans. In this review, first we describe common mouse models for behavioural analyses. As both genetic and environmental factors influence behavioural performance and need to be carefully evaluated in behavioural experiments, considerations for designing and interpretations of these experiments are subsequently discussed. Finally, common behavioural tests used to assess brain function are reviewed, and it is illustrated how behavioural tests are used to increase our understanding of the role of histaminergic neurotransmission in brain function. PMID- 16035959 TI - Chronic administration of venlafaxine fails to attenuate 5-HT1A receptor function at the level of receptor-G protein interaction. AB - In this study venlafaxine was administered to rats at a low, moderate or high dose; for comparison, the selective serotonin reuptake inhibitor (SSRI) sertraline and the tricyclic antidepressant (TCA) amitriptyline were also included. We evaluated, using quantitative autoradiography, the effect of these antidepressant treatments on [35S]GTPgammaS binding stimulated by the 5-HT1A receptor agonist 8-OH-DPAT, a measure of the capacity of 5-HT1A receptors to activate G proteins. Chronic administration of amitriptyline resulted in a marked increase in 5-HT1A receptor-stimulated [35S]GTPgammaS binding in the hippocampus which was accompanied by an increase in 5-HT1A receptor number. 5-HT1A receptor stimulated [35S]GTPgammaS binding in the hippocampus was also increased by chronic treatment with the highest dose of venlafaxine; 5-HT1A receptor number, however, was not significantly altered. In serotonergic cell body areas (i.e. dorsal and median raphe nuclei), 5-HT1A receptor-stimulated [35S]GTPgammaS binding was not altered by chronic administration of amitriptyline, sertraline or venlafaxine. Chronic TCA treatment does not desensitize somatodendritic 5-HT1A autoreceptor function. However, the lack of effect of chronic sertraline treatment on 5-HT1A receptor-stimulated [35S]GTPgammaS binding is in contrast to what has been observed previously following chronic administration of the SSRI fluoxetine, and suggests that different SSRIs may regulate somatodendritic 5-HT1A autoreceptor function differently depending on their pharmacology. Our data also suggest that the desensitization of somatodendritic 5-HT1A autoreceptors observed in electrophysiological studies following chronic venlafaxine administration is not at the level of receptor-G protein interaction. The hypothermic response in vivo to acute injection of 8-OH-DPAT was significantly attenuated following chronic treatment with venlafaxine or sertraline, but not amitriptyline. PMID- 16035958 TI - Chronic treatment with fluoxetine up-regulates cellular BDNF mRNA expression in rat dopaminergic regions. AB - During the last few years several studies have highlighted the possibility that major depression can be characterized by a general reduction in brain plasticity and an increased vulnerability under challenging situations. Such dysfunction may be the consequence of reduced expression and function of proteins important for neuroplasticity such as brain-derived neurotrophic factor (BDNF). On this basis, by using a sensitive non-radioactive in-situ hybridization, we evaluated the effects of a chronic treatment with fluoxetine on BDNF expression within rat dopaminergic regions. In fact, besides the well-established role of the hippocampus, increasing evidence indicates that other brain regions may be involved in the pathophysiology of depression and consequently be relevant for the therapeutic action of antidepressant drugs. Our results indicate that 3 wk of fluoxetine administration up-regulates BDNF mRNA levels selectively within structures belonging to the meso-cortico-limbic pathway. The expression of the neurotrophin is significantly increased in the ventral tegmental area, prefrontal cortex, and shell region of the nucleus accumbens, whereas no changes were detected in the substantia nigra and striatum. Moreover, in agreement with previous studies, fluoxetine increased BDNF mRNA levels in the hippocampus, an effect that was limited to the cell bodies without any change in its dendritic targeting. These data show that chronic treatment with fluoxetine increases BDNF gene expression not only in limbic areas but also in dopaminergic regions, suggesting that such an effect may contribute to improve the function of the dopaminergic system in depressed subjects. PMID- 16035961 TI - E-health and the Universitas 21 organization: 1. Global e-health through synergy. AB - The Universitas 21 (U21) organization funded a one-year project to examine global e-health. An e-health steering committee surveyed the opinions of e-health researchers at U21 member schools and conducted a literature review. Information about key themes was analysed and the findings were summarized. The steering committee recommended an eight-step strategy to establish a sustainable endeavour in global e-health. This included implementing a dissemination strategy within the U21 organization to engage a progressively larger community of faculty members and others, and translating e-health knowledge into global practice in those areas in which the U21 has special expertise. While the recommendations in the discussion paper are specific to the U21 organization, the e-health steering committee believes they can be generalized and applied to any globally minded educational or research institutions seeking to contribute to e-health. PMID- 16035962 TI - E-health and the Universitas 21 organization: 2. Telemedicine and underserved populations. AB - Telemedicine activities in underserved communities were reviewed as part of the Universitas 21 (U21) e-health project. A SWOT analysis (strengths, weaknesses, opportunities, threats) was conducted on 12 articles identified in a literature review, supplemented by expertise from U21 members. The analysis showed that threats include the reluctance of populations to use telemedicine services, and a general absence of infrastructure and resources to sustain them. Opportunities centre around potential research, including cost-effectiveness analyses and quantitative assessments of existing telemedicine services. The great strength of telemedicine is that it can improve access to health services among those most in need. However, its greatest weakness is the lack of evidence supporting its clinical and cost advantages relative to traditional services. This represents an important opportunity for research on telemedicine initiatives among underserved populations. PMID- 16035963 TI - E-health and the Universitas 21 organization: 3. Global policy. AB - There is an urgent need to develop global e-health policy in order both to facilitate and to manage the potential of e-health. As part of the Universitas 21 (U21) project in e-health, an evaluation of the status of global e-health policy was performed using a SWOT analysis (strengths, weaknesses, opportunities and threats). The analysis showed that the greatest threat to global e-health policy is the autonomous nature of domestic health-care systems. The greatest opportunity may be the prospect for nations and individuals to work together in establishing mechanisms necessary to offer health-care access through global e health--a new 'global public good'. Full integration of e-health into existing health-care systems could be achieved in both a practical and a policy sense through global e-health policy initiatives that facilitate integration across jurisdictions. There is a pressing need to resolve a range of e-health policy issues, and a concomitant need for research that will inform and support the process. A process that adopts a global approach is recommended. PMID- 16035964 TI - E-health and the Universitas 21 organization: 4. Professional portability. AB - Professional portability is the ease with which health-care professionals can move in person or virtually across barriers, and among and between jurisdictions, to transfer their knowledge, skills and care. As part of the Universitas 21 (U21) project on e-health, professional portability was examined using a SWOT analysis (strengths, weaknesses, opportunities and threats). The analysis showed that many factors hamper the development of global professional portability; on the other hand, the potential exists to substantially improve access to health care and its quality around the world. The study suggests that professional portability can be advanced in a number of ways. These include exploring policy, technology and medical training. The field of professional portability, while of considerable relevance to health and other professions, is undeveloped and is clearly an area that would benefit from discussion, research and global collaboration. PMID- 16035966 TI - Trans-telephonic monitoring of continuous haemodynamic measurements in heart failure patients. AB - The development of an implantable haemodynamic monitor (IHM) has made possible the home monitoring of a patient's central haemodynamic trends during daily living. We have evaluated the usability and transmission success of such a system over a 3.2-year period. Patients with an IHM were able to transmit the stored data every week to a Web server, where trend plots and tables of haemodynamic variables could be viewed by the health-care staff concerned. Data transfer was examined from August 2000 to November 2003. During this period, 148 patients had an IHM implanted. Over the study period, 7791 data transmissions were performed and an average of 286 downloads per month were sent to the Web server. In all, 86% of data transmissions were successful, although 10% required more than one attempt. The study demonstrated that telemonitoring of haemodynamic data from an IHM was feasible. A patient survey showed that the technology was user-friendly and that the training material provided sufficient information for patients and their families to install and use the transmission equipment at home. It also suggested that transmission success was independent of patient age or gender. PMID- 16035965 TI - A review of the first five years of the University of Arizona telepsychiatry programme. AB - We performed a retrospective review of the first five years of activity in the University of Arizona Telepsychiatry Programme. The programme began in 1998 as part of a telemedicine service for underserved rural areas. Over the first five years, 1086 teleconsultations were carried out for a total of 206 patients. There were 159 adult (77%) and 47 paediatric patients (23%). During the study period, the racial/ethnic diversity of the paediatric patients increased, as did the number of follow-up visits for adult patients (average 5.3/patient). These trends suggested that telepsychiatry became an integral part of the mental health service system at the six referring sites. Adult patients completed 81 satisfaction surveys (51%) and psychiatrists completed 47 (23%). The responses showed that providers and patients found telepsychiatry services satisfactory, although in a minority of teleconsultations (18% for providers and 17% for patients) equipment problems were distracting. The continued use of the services suggests that the telepsychiatry programme fills a service gap in rural areas of the state. PMID- 16035967 TI - Telemedicine in radiotherapy: a study exploring remote treatment planning, supervision and economics. AB - In January 2002, the departments of radiotherapy at the University Hospital of North Norway and the Norwegian Radium Hospital were connected through a 2 Mbit/s digital telecommunication line. The treatment planning systems at the two institutions were connected and videoconferencing units were installed. We explored the feasibility of remote treatment planning, supervision, second opinions and education. Tests involved two dummy cases and six patients. Remote simulation procedures were carried out for five patients. A cost-minimization analysis was performed. Treatment planning was not completely successful as the software could not handle plans including bolus or weighting between the fields. Remote supervision was possible. A common patient record and radiotherapy system, including digital imaging, digital prescription and approval forms and digital signature, were felt to be desirable. The threshold (break-even point) comparing the costs of telemedicine with those of transportation by air was 12 patients/year. Telemedicine in radiotherapy appears to be feasible, but some limitations must be overcome. PMID- 16035968 TI - Telemedicine in haemodialysis: a university department and two remote satellites linked together as one common workplace. AB - A common workplace was established between the renal unit at the University Hospital of North Norway and two satellite dialysis centres, in Alta and Hammerfest. A 2 Mbit/s ATM network was employed for IP-based videoconferencing. A common electronic medical record system and dialysis monitoring software were used. During an eight-month study period, nine patients were enrolled and 225 videoconferences were performed for daily visits and regular rounds. A bandwidth of 768 kbit/s was required for satisfactory teledialysis. Although technical (28%) and logistical problems (10%) were frequent, five hospitalizations and one third of the planned visiting rounds were avoided. An economic analysis showed that annual savings amounted to US$46,613, while annual costs were US$79,489. Despite the technical difficulties in about 30% of conferences, the nurses were satisfied with the videoconferencing system. Digital X-rays were communicated without problems. The pilot study indicates that satellite units may be incorporated into the daily management at the central institution by telemedicine. PMID- 16035969 TI - Evaluation of a realtime, remote monitoring telemedicine system using the Bluetooth protocol and a mobile phone network. AB - A generic, realtime wireless telemedicine system has been developed that uses the Bluetooth protocol and the general packet radio service for mobile phones. The system was tested on 10 healthy volunteers, by continuous monitoring of their electrocardiograms (ECGs). Under realistic conditions, the system had 96.5% uptime, a data throughput of 3.3 kbit/s, a mean packet error rate of 8.5x10(-3) packet/s and a mean packet loss rate of 8.2x10(-3) packet/s. During 24 h testing, the total average downtime was 66 min and 90% of the periods of downtime were of only 1-3 min duration. Less than 10% of the ECGs were of unacceptable quality. Thus, the generic telemedicine system showed high reliability and performance, and the design may provide a foundation for realtime monitoring in clinical practice, for example in cardiology. PMID- 16035970 TI - Using telemedicine to improve communication during paediatric resuscitations. PMID- 16035971 TI - Telephone consultations in general practice: areas for improvement. PMID- 16035972 TI - Australian nurses' perceptions of e-health. PMID- 16035974 TI - Introducing assistive technology into the existing homes of older people: feasibility, acceptability, costs and outcomes. AB - We examined how far, and at what cost, the housing stock could be modified to accommodate the assistive technology (AT) necessary to enable older people to remain in their own homes. A multidisciplinary team devised seven hypothetical user profiles for 10 case study areas, with five local authorities and five housing associations in England and Wales. Each profile was considered at two times, five years apart, with the users' functional abilities deteriorating in between. In addition, in-depth interviews were carried out with a sample of 67 older people in the case study areas about their use and experience of a wide range of AT. The interviews showed the need to listen to older people and that they welcomed AT when it addressed a perceived need. The results showed that the extent of adaptation required of buildings to accommodate a user's needs varied greatly. It was also found that there was confusion about the terminology of AT, including the idea of the 'smart house'. The study shows that the adaptability of the housing depends on a range of factors and costs. PMID- 16035975 TI - Experience-based guidelines for the implementation of telemedicine services. AB - Many telemedicine projects fail to survive beyond the funded research phase. A review of seven Scottish telemedicine services was conducted to identify successes and failures. Qualitative interviews were conducted with key individuals in each project. All projects were partly successful. The main reasons associated with partial failure were: the service was not needs-driven; there was no commitment to provide the service; there was no suitable exit strategy after research funding expired; there was poor communication; there was a lack of training; there were technical problems; work practices were not updated; the protocols for use were poor or non-existent. Based on this, guidelines that might improve the chances of success in future projects were drawn up. PMID- 16035976 TI - Implementing autonomy in a diabetes management system. AB - We have developed a speech-based telemedicine system which enables patients with hypertension and type 2 diabetes mellitus to send frequent, home-monitored health data via the telephone to the point of care. The decision support module in the system was tested using data from a cohort of 10 patients generated over a two year period. Results from the tests indicate that the system is effective in providing personalized feedback to the patient and in generating alerts for the clinical user. The work suggests that this method of care delivery is practical, informative, and may improve the efficiency of chronic health-care delivery by reducing costs and improving patient-physician communication between hospital visits. PMID- 16035977 TI - Wellness management through Web-based programmes. AB - We established a Web-based programme called the 'Wellness Online Program' or WOLP. The programme runs for six weeks. It aims to help individuals manage their own wellness regardless of geographical location. WOLP is based on a holistic approach to health and consists of six wellness dimensions: physical (exercise and diet), emotional, social, intellectual, spiritual and occupational. A total of 150 volunteers from the general public were recruited online for this study and data were collected at three intervals: at the beginning of the programme (week 1), mid-programme (week 3) and at the end of the programme (week 6). Ninety of the participants (60%) completed the six-week programme. Acceptance of WOLP, measured by the frequency of individual usage, increased from the start to the end of the programme. Overall personal wellness management improved after six weeks on the wellness programme (5-10%). The study shows that personal wellness management, which had been demonstrated in the narrower confines of the corporate sector, is also possible in the public domain. PMID- 16035978 TI - Wearable devices for telemedicine applications. AB - Wearable medical devices can provide both continuous monitoring and ubiquitous treatment. Challenges in this area include the need for a low-power/power-saving design to extend battery life and to reduce the size of the battery itself. This is followed by size and weight restrictions to meet patient expectations of what is 'wearable', the biocompatibility of all outer housings and the final assembly concept. Two examples of wearable medical device are described: a wrist-wearable telemedicine monitor for heart patients (AMON) and a generic belt-integrated computing platform for home and hospital use (QBIC). The electrocardiogram (ECG), the blood oxygen saturation (SpO2) sensor and the blood pressure meter of the AMON device were tested with 29 subjects. The sensors were found to be functional, but as expected the data processing algorithms will need some fine tuning. The prototype QBIC demonstrates a size reduction of 30-50% in relation to comparable devices. PMID- 16035979 TI - Cardiac event recording yields more diagnoses than 24-hour Holter monitoring in patients with palpitations. AB - Palpitation is a common symptom that sometimes results from a substantial cardiac arrhythmia. We compared the diagnostic yield of trans-telephonic event monitors with those of Holter monitoring in patients with intermittent palpitations. In all, 310 patients were randomly assigned to receive an event recorder or 24-hour Holter monitoring. Event recorders were used for seven days or until two recordings were obtained while symptoms occurred. The main end-point was an electrocardiogram (ECG) recorded during symptoms. The patients with palpitation recorded the one-lead ECG trace and sent it to a telemedicine call centre, where a nurse responded. There were 119 symptomatic patients in the event recorder group and 74 in the Holter group. The total costs were 6019 for event recording and 9605 for Holter monitoring. The average costs were 51 per symptomatic patient detected by event recorder monitoring and 130 per symptomatic patient detected by Holter monitoring. More patients therefore received a clear diagnosis, and more quickly, when using event recording than with Holter monitoring. For this reason, event recorders are preferable to Holter monitors for patients with palpitations. PMID- 16035980 TI - Effect of home-based telecardiology on chronic heart failure: costs and outcomes. AB - Chronic heart failure (CHF) remains a common cause of disability. We have investigated the use of home-based telecardiology (HBT) in CHF patients. Four hundred and twenty-six patients were enrolled in the study: 230 in the HBT group and 196 in the usual-care group. HBT consisted of trans-telephonic follow-up and electrocardiogram (ECG) monitoring, followed by visits from the paramedical and medical team. A one-lead ECG recording was transmitted to a receiving station, where a nurse was available for reporting and interactive teleconsultation. The patient could call the centre when assistance was required (tele-assistance), while the team could call the patient for scheduled appointments (telemonitoring). The one-year clinical outcomes showed that there was a significant reduction in rehospitalizations in the HBT group compared with the usual-care group (24% versus 34%, respectively). There was an increase in quality of life in the HBT group (mean Minnesota Living Questionnaire scores 29 and 23.5, respectively). The total costs were lower in the HBT group (107,494 and 140,874, respectively). The results suggest that a telecardiology service can detect and prevent clinical instability, reduce rehospitalization and lower the cost of managing CHF patients. PMID- 16035981 TI - Telecardiology: one-lead electrocardiogram monitoring and nurse triage in chronic heart failure. AB - We investigated a home-based intervention based on telecardiology in patients with chronic heart failure (CHF). Two hundred and thirty CHF patients, aged 59 years (SD 9), in stable condition and with optimized therapy were enrolled. The programme consisted of trans-telephonic follow-up and electrocardiogram (ECG) monitoring followed by visits from a paramedical and medical team. The patient could call the centre when required (tele-assistance), while the team could call the patient at prescheduled times (telemonitoring). During the first 12 months, there were 3767 calls (873 ad hoc and 2894 scheduled calls). There were 648 events, including 126 episodes of asymptomatic hypotension and 168 episodes which were not due to cardiological symptoms. No actions were taken by the nurse after 2417 calls (64%). A change in therapy was suggested after 418 calls, hospital admission in 62 patients, further investigations for 243 patients and a consultation with the general practitioner in 41 patients. A total of 2303 one lead ECG recordings were received (10 per patient); 126 recordings (6%) were diagnosed as pathological in comparison with the baseline one. The one-lead ECG recording was used for titration of beta-blockers in 79 patients (mean dosage 38 mg vs 42 mg, P<0.01). Home telenursing could be an important application of telemedicine and single-lead ECG recording seems to offer additional benefit in comparison with telephone follow-up alone. PMID- 16035982 TI - Teleradiology and picture archiving and communications systems: changed pattern of communication between clinicians and radiologists. AB - In a few years all radiology departments in Norwegian hospitals will have a picture archiving and communications system (PACS). This depends on telecommunications with great capacity for transmitting radiology images. The new technology questions the need for daily meetings between radiologists and clinicians, 'clinico-radiological conferences'. Qualitative interviews were performed with 23 resource persons experienced with problems related to PACS and teleradiology. The response rate was 91%. In all, 29% answered that the clinico radiological conferences could be abolished, 52% replied no and 19% replied both yes and no. The clinico-radiological conferences could be abolished for some clinical departments, but only after consultation between radiology and clinical departments. If the conferences are abolished, clinicians and radiologists may spend more time on treatment and interpretation, with a probable productivity gain. PMID- 16035983 TI - MPEG-21 as an access control tool for the National Health Service Care Records Service. AB - Since the launch of the National Health Service (NHS) Care Records Service with plans to share patient information across England, there has been an emphasis on the need for manageable access control methods. MPEG-21 is a structured file format which includes an Intellectual Property Management and Protection (IPMP) function using XML to present all digitally stored items in the patient record. Using DICreator software, patient records consisting of written text, audio recordings, non-X-ray digital imaging and video sequences were linked up successfully. Audio records were created using Talk-Back 2002 to standardize and optimize recording quality. The recorded reports were then linked and archived using iTunes. A key was used each time the file was displayed to secure access to confidential patient data. The building of the correct file structure could be monitored during the entire creation of the file. The results demonstrated the ability to ensure secure access of the MPEG-21 file by both health-care professionals and patients by use of different keys and a specific MPEG-21 browser. The study also showed that the enabling of IPMP will provide accurate audit trails to authenticate appropriate access to medical information. PMID- 16035984 TI - Health information seeking and its effect on the doctor-patient digital divide. AB - A survey of 224 individuals using SureStart services (for young families) within an inner-London area was complemented by qualitative data from five focus groups of parents and general practitioners in the same area. Descriptive and multivariate statistics were used to identify and describe discrete geographical districts with differing patterns of health information seeking. A geographically defined group of 'information hungry'/'online' health seekers was identified. This group contrasted with those acquiring information through 'assimilation' ('offline' information seekers). Qualitative data revealed the processes underpinning these characteristics and professional attitudes towards the Internet as a source of health information. PMID- 16035985 TI - Evaluation of the practical feasibility and acceptability of home monitoring in residential homes. AB - Three residential homes to the north-west of London, with their associated medical centres, were equipped with telemonitors to measure several variables, including seven-lead electrocardiogram, blood pressure, oxygen saturation, heart rate, temperature and respiration. The monitors could be operated by non-medical personnel. After recording, the data were transmitted via the Internet to a server. A total of 24 patients with a variety of chronic conditions were monitored regularly for a period of one year. Data transmission was found to be unreliable on occasions and was improved by extending the wireless network in the homes. Data access and presentation were considered acceptable, although suggestions for minor changes were made. No discernible impact on disease management, diagnosis or care was observed as a result of the monitoring. However, interviewees saw several potential benefits and patient acceptance was very good. The monitoring was found to be generally acceptable and feasibility was considered to have been largely proven. PMID- 16035986 TI - Interactive health communication applications for chronic disease: patient and carer perspectives. AB - Interactive health communication applications (IHCAs) may be particularly useful to patients and carers managing chronic disease. We have run eight focus groups with patients and two with carers involving a total of 40 participants. The focus groups were designed to seek patients' and carers' requirements of IHCAs and to identify the criteria they would use to assess them. Analysis revealed that many participants saw the value and potential of IHCAs. Even those with modest previous computer experience could use them with little tuition. The findings from this study have policy implications for the development of applications to maximize the potential benefit of IHCAs to patients and carers. PMID- 16035987 TI - Daily usage and efficiency of remote home monitoring in hypertensive patients over a one-year period. AB - We evaluated daily self-monitored blood pressure (BP) data collected over one year using remote home monitoring. Fifty treated, moderately hypertensive subjects (26 males, mean age 50 years; 17 females, mean age 54 years; seven exclusions) were recruited for the study in which semi-automatic arm-cuff BP measurement devices were used. The daily self-monitoring regimen had two phases of usage: one of initial enthusiasm (the first one to two months) followed by a phase of lower usage (89% versus 64%, P<0.01). Monitoring was missed more often (P < 0.01) during weekends (7.3 instances per patient) compared with workdays (5.0). Lack of motivation was not considered to be a major barrier. Approximately half of the study population was willing to continue the trial at the end of the one-year study. The occurrence of extreme BP values dropped significantly after the initial study month (P = 0.02). In conclusion, routine remote BP monitoring is capable of collecting consistent and accurate data, with sufficient sensitivity to reveal trends. PMID- 16035988 TI - Preferences of general practitioners regarding an application running on a personal digital assistant in acute stroke care. AB - An application was developed to optimize information exchange in acute stroke care, with which general practitioners (GPs) could consult hospital emergency units. However, it was difficult to obtain clear preferences from GPs regarding the functional requirements of the information to be transferred or the architecture of the application. Thirteen GPs volunteered to take part in the study. The GPs used a personal digital assistant in their daily work for a period of six weeks when visiting stroke patients during their evening, night and weekend shifts. A conjoint analysis was conducted to obtain the least and most preferred characteristics of the application, to facilitate implementation on a larger scale. The main outcome was that GPs preferred the decision-support facilities and the presence of information about the patient's medical history. PMID- 16035989 TI - A Web-based intervention used in general practice for people with excessive alcohol consumption. AB - Down Your Drink is a Web-based brief intervention for hazardous drinkers. We conducted a pilot study of its usefulness in general practice. Visitors to the Website were invited to complete the Fast Alcohol Screening Test and positive responders were offered a six-week interactive Web-based programme. There were 1319 registrations in the six-month study period. In all, 815 (62%) registrants completed week one, and 79 (6%) stayed with the programme until the end. Usage of the Website was distributed through the day, with the highest levels between 10:00 and 14:00. Two focus groups with general practitioners were conducted to explore the potential for using the site in primary-care settings. Website interventions for excessive drinkers therefore appear to be feasible and may prove a useful additional tool for intervention in general practice. PMID- 16035990 TI - A critical analysis of the literature on the Internet and consumer health information. AB - A critical review of the published literature investigating the Internet and consumer health information was undertaken in order to inform further research and policy. A qualitative, narrative method was used, consisting of a three-stage process of identification and collation, thematic coding, and critical analysis. This analysis identified five main themes in the research in this area: (1) the quality of online health information for consumers; (2) consumer use of the Internet for health information; (3) the effect of e-health on the practitioner patient relationship; (4) virtual communities and online social support and (5) the electronic delivery of information-based interventions. Analysis of these themes revealed more about the concerns of health professionals than about the effect of the Internet on users. Much of the existing work has concentrated on quantifying characteristics of the Internet: for example, measuring the quality of online information, or describing the numbers of users in different health care settings. There is a lack of qualitative research that explores how citizens are actually using the Internet for health care. PMID- 16035991 TI - Mobile phone technology in the management of asthma. AB - Peak flow monitoring is widely recommended as part of a self-management plan for asthma. We conducted an observational study using electronic peak flow monitoring and mobile phone technology in a UK general practice population over a nine-month period. Patients between 12 and 55 years of age who required treatment with regular inhaled steroids and (as needed) bronchodilators were recruited from nine general practices. Patients were included if their asthma was considered stable (i.e. no exacerbation in the previous three months). No therapeutic intervention was proposed. The primary outcome measure was compliance. In all, 69% of the 46 participants who filled in the post-study questionnaire were 'satisfied' or 'very satisfied' by the study, citing the ease of use and the increased autonomy and understanding of asthma as the main advantages. In total, 74% indicated that the system had helped to improve their ability to manage their symptoms. The most positive features of the telemedicine system were described as follows: increased awareness and information about asthma, improved ability to monitor/manage the condition with the feedback screens on the mobile phone and ease of use. PMID- 16035992 TI - Clinical trials and evaluation of a mobile, robotic tele-ultrasound system. AB - We have developed a robotic tele-ultrasound system (OTELO) that allows an expert to examine a distant patient by ultrasound. At the expert station, a sonographer controls a virtual probe. Movements are reproduced at the patient station, which may be several kilometres away, on a real probe held by a lightweight robot, which is positioned on the patient by a paramedic. Two medical teams tested the tele-ultrasound system at two different hospitals on a total of 52 patients. Except for some difficulties caused by particular conditions, the diagnosis obtained with the remote scanning system agreed in at least 80% of the cases with the diagnosis made by conventional scanning. The results demonstrate the feasibility and efficiency of the device. PMID- 16035993 TI - Telecare: perspectives on the changing role of patients and citizens. AB - We conducted a qualitative study in the UK to examine perspectives about telemedicine, telecare and e-health for the changing roles of patients and citizens. We believe there are important differences between these applications, but for simplicity, the term 'telecare' is used broadly here to include all three. In-depth, semistructured interviews were conducted between September 2002 and May 2004. Participants were 38 key informants from the UK, known to have involvement or interest in telecare: policy-makers, clinicians, technologists, health service managers, researchers and patient advocates. Interviews were conducted either in person or over the telephone, and were audio-recorded with participants' consent. The most frequently cited priorities for patients included accessibility to services, locations of care and quality of care, with some respondents emphasizing the importance of choice for the patient (in terms of appointments and ways of accessing services). However, telecare has implications for patients that go beyond issues of access. A major problem is that 'priorities' are assumed rather than based on an empirical understanding; moreover, for individual patients, preferences for particular forms of service delivery are likely to represent trade-offs between multiple priorities. The findings of the present study confirm that concepts of the patient's rights and responsibilities are changing with the increasing use of new technologies to deliver health care. PMID- 16035994 TI - Telemedicine in a child and adolescent mental health service: participants' acceptance and utilization. AB - Video-conferencing equipment was set up in Scotland in response to the increased pressure faced by the child and adolescent mental health services (CAMHS), and the need for specialist services to be accessible to, and harmonize with, 'mainstream' health services. Three sites were linked to the inpatient service in Edinburgh. Data were collected via questionnaires and diary logs. During a 24 month study, a total of 65 adolescents were admitted for inpatient care, of whom only five had their cases reviewed and monitored in a total of 20 teleconsultations. Adolescents and their carers involved in the study expressed great satisfaction with telemedicine and were keen to use it. Clinicians were resistant to telemedicine, with consequently low levels of utilization. Our results suggest that managers may be unwilling to re-allocate funding away from staffing, even where these costs are small and represent considerable improvements in the process of care for patients. Widespread integration of telemedicine to CAMHS is likely to be hard to achieve. PMID- 16035995 TI - A Web-based mobile asthma management system. AB - Continuous recording of daily symptoms constitutes an effective means of managing asthma patients. Daily management reduces the costs associated with hospitalization and improves the quality of patient care. We have implemented a Web-based mobile asthma management system. We used a pocket PC, mobile phone and desktop computer. The recorded items and individualized prescriptions were structured using Extensible Mark-up Language (XML) DTD (Data Type Definition). The mobile Web form was automatically adjusted to fit the different display resolutions of the terminal devices. The system provided reliable exchange of all relevant information between a doctor and the asthma patient using wireless mobile transmission. E-mail and Short Messaging Service (SMS) were used to send messages to patients, for example in the case of an automatically determined patient alert. Patients could obtain customized instructions according to their daily personal symptoms, peak expiratory flow (PEF), medications and activity restriction. The daily graph of PEF and the graphs of symptoms and medication were particularly useful for asthma patient control and self-awareness of the progress of the disease. PMID- 16035996 TI - Attitudes of young people with diabetes to an Internet-based virtual clinic. AB - We assessed the feasibility of developing a virtual diabetes clinic for young people, using the Internet. The proposed content of the site would be based on self-efficacy theory, aiming to develop confidence in self-management of diabetes. A questionnaire about the proposed Website was delivered to 72 patients who attended a young person's outpatient clinic in a district hospital. Thirty nine replies were received, a response rate of 54%. The mean age of the respondents was 13 years. A positive attitude to the potential clinic was reported by 95% of respondents. The items rated as most useful were: quick and easy access to up-to-date information about diabetes; the opportunity to ask an expert; good graphics; easy navigation; interactivity. Responses to open questions indicated that 24-hour access and anonymity in asking questions were also valued. A virtual clinic appears to be a possible method of health-care delivery to young people with diabetes. PMID- 16035997 TI - Misconceptions surrounding videoconferencing. AB - We carried out a study to assess the level of training required by hospital staff to operate a videoconferencing system. Fifty members of hospital staff, who had no previous videoconferencing experience, were studied. When using simple pictorial instructions, they took on average just under 6 min to connect successfully to a remote site. Male subjects connected faster (average 309 s) than female subjects (385 s; P < 0.01). Subjects of 40 years or younger connected faster (average 311 s) than those over 40 years (average 398 s; P < 0.02). Only three individuals (6%) required supplementary help via the telephone. Dedicated training may not be necessary when simple pictorial instructions are available to the operator. PMID- 16035998 TI - Role flexibility among telemedicine service providers in the north-west and west of Ireland. AB - A recent review of telemedicine services in the north-west and west of Ireland identified 11 telemedicine services, most of which were in the early stages of implementation. A qualitative approach was used to review them. Semistructured interviews were conducted with a multidisciplinary group of primary- and secondary-care providers (n = 21) who were involved with either synchronous or asynchronous telemedicine services. Data were analysed according to the principles of framework analysis. Participants described the ways in which they were flexible about their workload, professional identities and roles to facilitate the organization and delivery of telemedicine services, and to ensure that services ran smoothly. While the positive effect of product champions and members of a wider supportive network or alliance on the conception and development of telemedicine services must be acknowledged, questions remain about associated long-term implementation and sustainability. PMID- 16035999 TI - An online survey of nurses' perceptions, knowledge and expectations of the National Health Service modernization programme. AB - We conducted an online survey to investigate nurses' perceptions, knowledge and expectations of the National Health Service (NHS) modernization programme in the UK. The questionnaire was available for 28 days via the Website of the Royal College of Nursing. The questionnaire was completed by 2020 nurses, midwives and health visitors working in all sectors of the health service in a wide range of specialties and environments of care. Less than one-quarter of respondents felt that they had adequate information about NHS information technology (IT) developments. In all, 528 (26%) said this was the first they had heard of the initiatives. Only 383 respondents (19%) felt adequately informed about the development of electronic health records; 470 (23%) felt inadequately informed and 456 (23%) had only heard something about it. The findings of this survey suggest that nursing staff are not widely aware of current IT plans and programmes in the NHS. They suggest that nurses also lack confidence in using advanced IT, which is compounded by lack of training. PMID- 16036000 TI - Principles for telemedicine and telecare: the perspective of a citizens' panel. AB - Ten delegates at the conference Voluntarism, Health and Social Care were recruited to form a panel of citizens to debate and offer direction for the future of technologically mediated health care. The panel suggested various principles for the development of telemedicine and telecare, concerning: patients, users and carers, approach to service delivery, research and knowledge, and conditions of use. Many of the principles echoed the founding values of the National Health Service, yet have arguably been absent from both policy pronouncements and the telemedicine literature, which largely views new health technologies themselves as 'value free', i.e. developed untouched by social and political relations. A programme of citizens' panels should be developed so that an informed debate can take place about the development of telemedicine and telecare, to underpin policy and practice. PMID- 16036001 TI - Using telemedicine to provide post-discharge support for patients with spinal cord injuries. AB - We are conducting a randomized controlled trial of telemedicine with patients with spinal cord injuries in their own homes. Internet videoconferencing is used at a bandwidth of 128 kbit/s. Data collection began in March 2004. Twelve patients had entered the study by August 2004, but none had completed it. Preliminary results in one case suggest that telemedicine provided various benefits: (1) the patient received advice he would probably not have solicited; (2) it enabled an expert to view the entry site of a pin in the patient's halo brace, to determine whether the general practitioner should be contacted to arrange a swab; (3) it made it easier for the interviewer to understand family interactions during the session. Telemedicine offers an additional tool in the care of geographically widespread outpatients. PMID- 16036002 TI - Improved access to subspecialist diabetes care by telemedicine: cost savings and care measures in the first two years of the FITE diabetes project. AB - We have used telemedicine clinics supplemented by online education to provide effective care for children with diabetes. Before the programme began, the mean interval between visits was 149 days; in year 1 of the programme it was 98 days, and in year 2 it was 89 days. Before the programme, there were on average 13 hospitalizations a year (47 days) and this decreased to 3.5 hospitalizations a year (5.5 days). Emergency department visits decreased from 8 to 2.5 per year. On 10 occasions after the programme started, ketosis was managed by telephone intervention alone, relying on family-initiated calls. Over 90% of patients and family members expressed satisfaction with the telemedicine service and wished to continue using it. In all, 95% felt little self-consciousness. Over 90% felt their privacy was respected. The programme saved US dollar 27,860 per year. The present study demonstrated improved access to specialized health care via telemedicine in combination with online education improved health status and reduced costs by reducing hospitalizations and emergency department visits. PMID- 16036003 TI - School nurse, family and provider connectivity in the FITE diabetes project. AB - The Florida Initiative in Telehealth and Education (FITE) diabetes project includes a system of remote blood glucose monitoring and online education for school personnel, families and providers. Forty-four patients with diabetes (100% of patients), six caregivers, six case managers and 18 school nurses were provided with secure email access, allowing blood glucose and other data transfer. In all, 50% of school nurses and 100% of case managers completed educational modules on the FITE Website. Over 90% of patients and all school nurses received equipment for transmitting blood glucose data to their computers. The data were discussed during clinic appointments. Inclusion of previously unavailable data from school nurses contributed to fine-tuning the diabetes management regimen. Those patients, families and school nurses who chose to transmit blood glucose data and participate in online education expressed satisfaction with the technology, the process and the improved communication. PMID- 16036004 TI - Mobile telemetry for pre-hospital thrombolysis: problems and solutions. AB - In the Angus region of Scotland, we have used mobile telemetry to facilitate pre hospital thrombolysis by paramedic staff. An initial survey demonstrated that connection could be achieved in all but three locations. In the first year of operation, 229 contacts were received. Communication between the ambulance and the base station failed on four occasions (2%). Problems with transmission of an electrocardiogram (ECG) were encountered on 37 occasions (16%). The median time for acquisition and transmission of an ECG was 22 min. This compares with a median time of 59 min for first ECG in a control group from similar locations, who were assessed in hospital. Telemetry offers essential back-up to paramedics adopting a challenging and extended role. Strategies can be developed to deal with signal strength and equipment failure. PMID- 16036005 TI - A Brazilian model of distance education in physical medicine and rehabilitation based on videoconferencing and Internet learning. AB - We have developed an integrated educational model in rehabilitation mediated by technology. Three teams of professionals worked to implement the interactive model based on videoconferencing, use of the Internet and three-dimensional (3-D) animated models. Two courses were created: amputee rehabilitation and back pain. Each course was divided into four phases: (1) a first videoconference; (2) Internet-based learning; (3) a second videoconference (workshop); (4) an Internet discussion list. The Internet-based learning modules were divided into topics by multiple-choice questions. Multisite videoconferences were used to connect the remote sites. Eleven animated 3-D models were created to help the teaching process. Each course had 11 modules, and each module required up to 2 h to be completed. There were 136 participants on the two courses. None of the participants, including the teachers, had had any previous experience with Internet-based learning and videoconferencing. The integrated educational model has great potential in a country the size of Brazil, where there may be difficulties in travel for patients with disabilities and for health-care professionals. PMID- 16036006 TI - The efficacy of telemedicine for ophthalmology triage by a general practitioner. AB - Although there are enough ophthalmologists for the Brazilian population, they are not evenly distributed throughout the country. Tele-ophthalmology may therefore be a useful tool. We have examined the feasibility of ophthalmology triage, performed by a general practitioner (GP) with remote support from an ophthalmologist. Forty patients with a variety of external and internal eye disorders were examined by the GP and also reassessed by an ophthalmologist, face to face, and then remotely by another ophthalmologist. There was agreement in 95% of the diagnoses between face-to-face and distant evaluation. The use of a digital camera and slit-lamp allowed greater accuracy of telediagnosis than the use of a digital camera alone. The GP would have referred 36 patients to an ophthalmologist, while both the local and the remote ophthalmologist saw the need for referral in 31 cases, i.e. assessment by tele-ophthalmology resulted in a 14% decrease in referrals. GP triage therefore appears to be feasible after appropriate training. PMID- 16036007 TI - Telemedicine-assisted home support for patients with advanced chronic obstructive pulmonary disease: preliminary results after nine-month follow-up. AB - Eighteen well motivated patients with advanced chronic obstructive pulmonary disease, who had had at least four hospitalizations during the previous two years, were included in a home-based telemedicine study. A visiting nurse was equipped with a case containing a laptop computer and a number of medical devices, including an electrocardiogram recorder, spirometer, oximeter and blood pressure monitor. It also contained a videoconference camera, for realtime audiovisual connection with the hospital using the patient's TV set. A single ISDN line (128 kbit/s) was installed in each house before the study began. After nine months, there was a decrease in hospitalizations, emergency department visits and use of health services. The patient's disease knowledge and self management also improved. It seems likely that adopting telemedicine in everyday clinical practice could substantially improve the care of chronically ill patients. PMID- 16036008 TI - Measurements of the clinical competence of doctors and nurses to process telemedicine referrals for burns patients. AB - Simulated telemedicine referrals were used to test the ability of groups of clinicians to manage telemedicine referrals of patients with burn injuries. Sixty one participants recorded burn depth from digital images using a four-point scale. The definitive ('gold standard') diagnoses were based on a review by an experienced consultant burns surgeon. Sixty clinical cases that reflected the routine referrals to a specialist burns service were used for the study. The mean kappa scores for the participants ranged from 0.33 to 0.58, indicating poor to good agreement. The scores for the groups all had a similar pattern, with more experienced staff scoring higher than junior staff. The doctors and nurses specializing in burns had higher scores than the general surgical nurses. PMID- 16036009 TI - Supporting families of critically ill children at home using videoconferencing. AB - Home videoconferencing links for families of children recently discharged with complex congenital heart disease may be useful in monitoring potentially unstable patients. A randomized controlled trial was carried out comparing home videoconferencing with telephone contact. Patients were randomized to an interventional videoconferencing group (n = 14), or to one of two control groups: the first (n = 9) received the same ad hoc telephone support that was available to all patients; the second group (n = 13) received regular telephone calls with the same protocol as those in the videoconferencing group. The results from the trial are still being analysed. Our experience with commercial cable modem transmission quickly showed that this is an unsuitable modality. Preliminary results with ISDN videoconferencing are encouraging. Initial results and feedback from families strongly suggest that videoconferencing provides significant benefits over telephone follow-up. PMID- 16036010 TI - Role of telecardiology in the assessment of angina in patients with recent acute coronary syndrome. AB - We compared two models of assistance (telecardiology versus usual care) for patients discharged after acute coronary syndrome (ACS), in the assessment of angina. Two hundred patients were randomized into two groups at discharge for ACS: Group A to telecardiology and Group B to usual care. Early hospital readmission (in the first month) occurred in 16 patients (seven in Group A and nine in Group B). Six of Group A were readmitted for a cardiac cause (non-cardiac in one). Angina was the only cardiac cause. Five of the Group B patients were readmitted for a cardiac cause (non-cardiac in four). The results of the present study emphasize that patients with ACS suffer from a definite rate of cardiac symptoms within the first month (63%). Angina occurs more frequently within the first two weeks (68% of cases). Telecardiology slightly reduces hospital readmissions (telecardiology 44% versus usual care 56%), but better identifies true angina. PMID- 16036011 TI - Patient and provider perspectives on home telecare: preliminary results from a randomized controlled trial. AB - A randomized controlled trial of home telecare for the management of acute exacerbations of chronic obstructive pulmonary disease has been undertaken in the north-west of England. A videophone was used that communicates via the ordinary telephone network. The intervention period for each participant was two weeks. Participants in the telecare arm of the trial were asked to complete logbooks to record their experiences of each telecare encounter. A simple, self-completed, 10 item questionnaire was used that consisted of a Likert scale, ranging from 1 (totally disagree) to 5 (totally agree). Fourteen nurses completed 150 logbooks and 22 patients completed 145 logbooks. These results demonstrate significant differences in perception between patients and their health-care providers with regard to telecare encounters across all the domains addressed. Participating patients consistently demonstrated more positive views of the telecare encounters than their healthcare providers. PMID- 16036012 TI - Unattended home sleep studies for the evaluation of suspected obstructive sleep apnoea syndrome in children. AB - Children suspected of obstructive sleep apnoea syndrome (OSAS) are traditionally investigated in hospital with overnight sleep studies, and single-channel pulse oximetry is the commonest form of initial study. A prospective study was conducted comparing unsupervised sleep studies, using a Nellcor N-395 pulse oximeter, performed in hospital and at home. Two hundred and eleven patient studies were completed in 18 months, 158 in hospital and 53 at home. The median age of patients was four years in both groups. The mean study duration was 9 h 5 min in hospital and 9 h 12 min at home (P = 0.75). The mean time in pulse search was 0.4% (SD = 0.7) in hospital and 0.2% (SD = 0.4) at home (P = 0.008). The number of failed studies was 96 of 254 (38%) in hospital and three of 56 (5%) at home (P < 0.001). Overall, 149 hospital bed days were saved (at pounds 690 per bed day), representing pounds 68,540 saved annually. The results show that unattended home sleep studies in the form of overnight pulse oximetry can be performed to a standard better than hospital studies, where local community nursing facilities exist. PMID- 16036013 TI - Developing e-learning for interprofessional education. AB - An evidence-based, interprofessional educational course involving first-year undergraduate students studying medicine, nursing, physiotherapy and occupational therapy has been piloted at the University of Liverpool. Part of the content was developed in an online format. To capture the development process and the e learning writing experience, a focus group was arranged for the content writers. The session was audio-recorded and tapes were transcribed. All the data were subjected to thematic analysis. Twenty-three sub-themes were identified in the 72 comments. These were grouped under six themes, corresponding to the developmental stages of e-learning. The highest number of comments fell under the theme of preparation, followed by content development, evaluation, general design and structure, and finally delivery. Team working contributed to the success of the writing process, reflecting the theme of working inter-professionally. PMID- 16036014 TI - Adapting the Roter interaction analysis system for telemedicine: lessons from four specialty clinics. AB - Four specialists (a child psychiatrist, an oncologist, a cardiologist and a rheumatologist) conducted telemedicine clinics using videoconferencing at a bandwidth of 128-384 kbit/s. The consultations were videorecorded. The coded interactions from the first two patients recruited from each of the four telemedicine clinics were analysed. Tapes were coded by two raters. We adapted the Roter interaction analysis system (RIAS) for the telemedicine context. Utterances were coded into socio-emotional and task-focused categories. There were 1055 utterances in total. Providers generated significantly more utterances across categories than patients. In the patient-provider interactions, only 2% of the total utterances related to the technology. The predominance of socio emotional utterances compared with task-focused utterances for providers was contrary to our expectations. Further studies are required to establish the reliability of the adapted RIAS measure and to increase understanding of telemedicine communication patterns. PMID- 16036015 TI - Diffusion theory and telemedicine adoption by Kansas health-care providers: critical factors in telemedicine adoption for improved patient access. AB - Twenty counties in Kansas were randomly selected from those designated as rural on the basis of their populations. A sample of 356 physicians and physicians' assistants in these counties was chosen. A postal survey was sent to the identified providers up to three times. One hundred and eighty-six of the questionnaires were returned (a response rate of 52%). In all, 76% of the respondents were physicians, 76% were men and 42% were family practitioners. Practitioners were classified as adopters or non-adopters of telemedicine, based on their report of whether they had ever referred one or more patients for a health-care consultation via telemedicine. Of the 167 participants who marked this item, 30 (18%) were adopters and 137 (82%) were non-adopters. Among the adopters, 16 (53%) said that they expected to use telemedicine with about the same frequency or more often in the future. In contrast, 61 (45%) non-adopters reported that they did not expect to refer patients by telemedicine in the future and 51 (37%) were unsure. Neither age (r = 0.16, P = 0.44) nor gender (chi2 = 2.35, P = 0.13) was related to the adoption variable or the number of referrals made to telemedicine clinics. The results suggest that adopters and non-adopters of telemedicine perceive its value very differently, and that an opportunity exists to promote the concept to non-adopters more effectively. PMID- 16036018 TI - Structural basis of ventricular remodeling: role of the myocyte. AB - Structural remodeling plays a major role in the progression of various heart diseases to congestive heart failure (CHF). Major contributors to this remodeling process in the heart include alterations in myocyte shape, myocyte number, and extracellular matrix. However, it is unclear as to which of these changes is most critical in the development of CHF, and this may vary by etiology. Myocyte shape alterations largely underlie the increase in chamber diameter/wall thickness characteristic of CHF. This review mainly focuses on the role of myocyte shape in ventricular remodeling. Several signaling molecules have been implicated in this process. As we learn more about the components of myocardial remodeling, new strategies to combat the progression of heart disease should arise. PMID- 16036019 TI - Pharmacologic effects on cardiac remodeling. AB - Clinical research during the past several decades has shown the importance of cardiac remodeling as a basic mechanism in the progression of heart failure. Changes in cardiac remodeling have high concordance with long-term clinical outcomes. Cardiac remodeling should be regarded as a primary target for treatment and also can serve as a reliable surrogate for clinical outcomes. Neurohormonal blockade with combination treatments that inhibit the renin-angiotensin aldosterone and sympathetic systems has proven effective in improving cardiac remodeling and clinical outcomes. Such treatment should be standard therapy for patients with left ventricular dysfunction after myocardial infarction and patients with chronic heart failure. PMID- 16036020 TI - Mechanical approaches to alter remodeling. AB - Regression of pathologic cardiac hypertrophy and dilation, so-called reverse remodeling, has emerged as an important therapeutic target in the treatment of dilated cardiomyopathies. Although pharmacologic therapies may promote regression of pathologic remodeling, the magnitude of reverse remodeling is usually small. In contrast, reverse remodeling associated with cardiovascular devices, as highlighted in this review, often has been more rapid and reliable. For example, circulatory support with a left ventricular assist device produces the dramatic reverse remodeling in severely diseased hearts and typically provides myocardial tissue samples to generate new insights into the basic biology of reverse remodeling. Alternatively, multisite ventricular pacing to improve the synchrony of ventricular contraction has demonstrated clinical efficacy that includes the ability to reduce cardiac dilation and hypertrophy, and improvements in symptoms and functional capacity. Passive cardiac support devices comprise another promising strategy to prevent or reverse detrimental cardiac remodeling in patients with dilated cardiomyopathies. PMID- 16036021 TI - Surgical ventricular restoration and other surgical approaches to heart failure. AB - Historically, few patients with ischemic congestive heart failure (CHF) have been considered for cardiac surgical intervention unless there was an obvious need for coronary revascularization or valve repair. New surgical procedures and non mechanical assist devices are being used and tested in patients with end-stage CHF. We report on The Ohio State University Medical Center's early involvement in the international and multi-institutional Surgical Treatment for Ischemic Heart Failure (STICH) trial, which is evaluating the value of coronary artery bypass in patients with ischemic CHF as compared to medical therapies alone, and whether surgical ventricular restoration (SVR) offers additional benefit to patients with dilated hearts undergoing revascularization. Beyond standard coronary revascularization and SVR, new surgically deployed devices that attempt to augment ventricular performance by direct restraint of left ventricular dilatation or by reducing ventricular wall stress through altering ventricular shape are reviewed. The growing clinical and experimental experience with cellular cardiomyoplasty (in particular, autologous skeletal myoblast and adult derived stem transplantation) also is reviewed. This review is intended to express the institutional insights of the authors, who have been involved in clinical trials and basic science research in each of these areas. PMID- 16036022 TI - Cardiac resynchronization therapy--how big of a miracle? AB - This review examines the clinical benefit of cardiac resynchronization therapy (CRT) in patients with severe heart failure symptoms and interventricular conduction delay. The magnitude of CRT effects on exercise duration, quality of life, and reversal of adverse remodeling are compared to angiotensin-converting enzyme (ACE) inhibitor therapy and combined beta-blocker/ACE inhibitor therapy. Overall, CRT delivers substantial improvement in clinical well-being to an extent similar to or greater than what would be expected with neurohormonal intervention alone. PMID- 16036023 TI - Ventricular assist devices: destination therapy or just another stop on the road? AB - Ventricular assist devices have attained center-stage recognition with the evolution of the concept of destination therapy, designed as an alternative to cardiac transplantation. However, mechanically induced hemodynamic restoration is accompanied by cellular and biochemical effects that may potentially mediate cardiac recovery. This has raised the fundamental question of whether mechanical assistance is a "failure to grave" option or simply a stop on the road to recovery. The concept of destination therapy as an alternative to transplantation was validated in the Randomized Evaluation of Mechanical Assistance in the Treatment of Congestive Heart Failure trial, but gaps in translating this information to the clinical realm exist because of device limitations of complications and durability. The field of mechanical assistance is progressing rapidly with the introduction of smaller devices that are more durable and have less risk for infection or hematologic aberrancies, which should allow this option to meaningfully enter the clinical arena. PMID- 16036024 TI - Interactions between the sympathetic nervous system and the RAAS in heart failure. AB - Therapy for heart failure caused by left ventricular systolic dysfunction is based on interference with maladaptive activation of the sympathetic nervous system (SNS) and the renin-angiotensin-aldosterone system (RAAS). Agents that block beta-adrenergic receptors, decrease angiotensin-II formation, and antagonize the effects of angiotensin II and aldosterone have been shown to improve morbidity and mortality in this syndrome. Therefore, from a theoretical point of view, it would be desirable to actually diminish the degree of overactivity of these two homeostatic systems. There are compelling physiologic arguments and much experimental data to suggest that beta-adrenergic blockade may diminish activity of the RAAS. Conversely, angiotensin-converting enzyme inhibitors, angiotensin-II antagonists, and aldosterone antagonists may diminish activity of the SNS. Some clinical trials data may be interpreted in a fashion that suggests that part of the benefit of interfering with each system may relate to diminishing activity of the other. If true, combined therapy may lead to a virtuous cycle in which diminishing the adverse effects of each individual system is combined with reduced activity of the other. Such a cycle may be one factor underlying the impressive clinical results of recent neurohormonally based therapeutic trials and reinforces the need to look beyond acute hemodynamic effects of therapeutic agents when assessing their long-term impact in heart failure. PMID- 16036025 TI - Efficacy of aldosterone receptor antagonism in heart failure: potential mechanisms. AB - Results of the Randomized Aldactone Evaluation Study and the Eplerenone Post acute Myocardial Infarction Heart Failure Efficacy and Survival Study indicate aldosterone receptor antagonism, together with angiotensin-converting enzyme inhibition and loop diuretics, is a most effective strategy in reducing risk for all-cause and cardiovascular-related mortality and morbidity in patients with symptomatic heart failure. Responsible mechanisms are likely multifactoral. As a circulating hormone, aldosterone has well-known endocrine properties that contribute to the pathophysiology of congestive heart failure. This includes Na+ resorption at the expense of K+ excretion in such tissues as kidneys, colon, sweat, and salivary glands. Mg2+ excretion at these sites is likewise enhanced by aldosterone, whereas adrenal aldosterone secretion is regulated by extracellular Mg2+. Other endocrine actions of aldosterone receptor-ligand binding include: a reduction in biologically active cytosolic-free Mg2+, with intracellular Ca2+ loading in nonepithelial cells such as peripheral blood mononuclear cells; its influence on endothelial cell function; and its central actions, including the choroid plexus, activity of the hypothalamic paraventricular nucleus, and autonomic nervous system. De novo generation of aldosterone within the cardiovasculature is recognized and findings suggest its auto/paracrine properties contribute to tissue repair. Each of these actions is interrupted by aldosterone receptor antagonism and therefore may contribute to its salutary response in heart failure. PMID- 16036026 TI - Inhibition of RAAS--when is it too much? AB - Deactivation of the renin-angiotensin-aldosterone system (RAAS) is clearly beneficial in patients with recent myocardial infarction and chronic heart failure. Most of the experience with deactivation of the RAAS has been collected in placebo-controlled randomized trials of angiotensin- converting enzyme inhibition (ACEI). The hypothesis that angiotensin receptor blockade may be a better approach to deactivate the RAAS has not survived the test of time. Despite the extensive experience with ACEI and aldosterone receptor blockade in patients with recent myocardial infarction and chronic heart failure, several issues remain unanswered. These are addressed in this review. PMID- 16036027 TI - How should COMET influence heart failure practice? AB - Much clinical experience has led us to assume that the administration of a b blocker, regardless of dose or frequency, would produce similar mortality benefits in patients with heart failure. The results from the recently published Carvedilol or Metoprolol European Trial (COMET), which found greater benefit of carvedilol than immediate-release metoprolol on mortality, clearly demonstrated this is not true. In heart failure, the COMET results strongly support the use of beta-blockers that have proven effective in large-scale clinical trials. The primary disagreement regarding COMET concerns the explanation of the efficacy difference between the two b-blockers tested. Pharmacodynamic considerations and hemodynamic data from the COMET trial itself suggest there were unequal degrees of b1-blockade between patients receiving carvedilol and immediate-release metoprolol. Failure to achieve a similar degree of b1-receptor blockade in the two groups prevents conclusions regarding the potential incremental benefits of selective versus nonselective adrenergic blockade. Further studies are needed to determine whether there are additional clinical benefits from the inhibition of adrenergic receptors beyond the proven benefits of b1-blockade. PMID- 16036028 TI - Initiation and use of beta-blockers in class IV heart failure. AB - Treatment of patients with heart failure caused by left ventricular systolic dysfunction using b-adrenergic receptor antagonists (or b-blockers) results in improvements in symptoms, hemodynamics, left ventricular remodeling, morbidity, and mortality. Most patients studied in prospective, randomized placebo controlled trials have had New York Heart Association (NYHA) functional class II or III symptoms. The efficacy of b-blockers in treating NYHA class IV patients is not as well-established. This review summarizes the published experience regarding the use of b-blockers in patients with advanced heart failure. Although treatment requires considerable care, the data support attempts at initiation of b-blockers in this group of patients. PMID- 16036029 TI - Beta-blockers in heart failure: is more better? AB - Therapies that modulate the sympathetic nervous system and renin-angiotensin aldosterone system reduce morbidity and mortality in patients with heart failure. However, they are grossly underused in clinical practice; when used, the doses prescribed are substantially smaller than the target doses used in the large scale studies that established their utility. Whether these suboptimal doses are as effective in reducing morbidity and mortality is largely unknown. This review focuses on the relationship between the dose of b-blockers and their effect on clinical outcomes. Because direct dose comparisons of b-blockers are limited, we draw upon a broader spectrum of clinical trials across the cardiovascular continuum that involved neurohormonal modulators to address the question, "Is more better?" PMID- 16036030 TI - Comprehensive adrenergic receptor blockade with carvedilol is superior to beta-1 selective blockade with metoprolol in patients with heart failure: COMET. AB - The Carvedilol or Metoprolol European Trial (COMET) compared the effects of a comprehensive adrenergic antagonist, carvedilol (target dosage 25 mg twice daily), with a beta-1-selective agent, metoprolol tartrate (target dosage 50 mg twice daily), in 3029 patients with chronic heart failure caused by left ventricular systolic dysfunction. The study showed that, compared with metoprolol, long-term treatment with carvedilol exerted a substantially greater reduction in mortality and led to improvement in well-being in these patients. The superiority of carvedilol over metoprolol is readily explained by differences in their pharmacologic profiles. There is no evidence, within the trial or from other sources, that the relative dose of each agent or their formulation explained the observed difference. The result of COMET is entirely consistent with the results of previous placebo-controlled trials of beta-1-selective blockers and carvedilol. PMID- 16036031 TI - The changing epidemiology of sudden death in heart failure. AB - Worsening and progressive heart failure has been the most common mode of death in patients with heart failure. With the development of drugs that primarily have modulation of the renin-angiotensin and sympathetic nervous systems as their mechanism, relative importance of progressive heart failure has decreased. Sudden death has become a larger issue and the dominant mode of death in heart failure. This change requires the clinician to redirect therapy to the prevention and treatment of sudden death. PMID- 16036032 TI - The molecular basis of cardiac arrhythmias in patients with cardiomyopathy. AB - Cardiac arrhythmias are a leading cause of mortality and morbidity in Western society. In some specific instances, these arrhythmias are caused by abnormalities of cardiac ion channels, such as sodium, calcium, and potassium channels, which carry ionic currents and are fundamental determinants of cardiac excitability. Abnormalities of these ion channels are attributed to mutations in the genes encoding the channel protein and cause altered function of channels, which can predispose to arrhythmias. During heart failure, many channels also malfunction because of altered expression, resulting in lethal arrhythmias. PMID- 16036033 TI - Current role of device therapy to reduce sudden cardiac death in heart failure. AB - Sudden cardiac death (SCD) continues to be a major contributor to mortality in patients with heart failure (HF) despite recent advances in medical therapy. Device therapy, including the implantable cardioverter defibrillator (ICD) and cardiac resynchronization therapy (CRT), serves as an adjunct in reducing HF mortality. Several clinical trials support the prophylactic use of the ICD in reducing mortality in certain HF populations and have established the clinical benefits of CRT in advanced HF. More recently, the Comparison of Medical Therapy Pacing and Defibrillation in Heart Failure trial was the first study to demonstrate a survival benefit of CRT alone or in conjunction with an ICD. This article reviews the most pertinent data regarding the role of device therapy in reducing SCD in HF and addresses future challenges faced by device manufacturers and clinicians. PMID- 16036034 TI - Cardiorenal syndrome in decompensated heart failure: prognostic and therapeutic implications. AB - Various lines of evidence implicate impaired renal function as an important prognostic indicator in patients with congestive heart failure (CHF). Conventional diuretics may aggravate renal dysfunction and can result in neurohumoral activation. Evolving new therapeutic strategies that enhance renal function include administration of B-type natriuretic peptide, adenosine and vasopressin antagonists, and ultrafiltration methods. Prospective studies are needed to evaluate whether these new renal-enhancing strategies will improve patient outcome in CHF. PMID- 16036035 TI - Quality of care and outcomes in acute decompensated heart failure: The ADHERE Registry. AB - Despite 1 million or more annual hospitalizations, the management of acute decompensated heart failure remains largely empiric. Data are lacking regarding patient characteristics, usual comorbidities, commonly used treatment strategies, clinical outcomes, and adherence to recently published quality indicators. The Acute Decompensated Heart Failure National Registry (ADHERE(R)) has been established to capture data regarding the presentation, management, and expected outcomes of acute decompensated heart failure. The size of the registry and its web-based platform allow for evaluation of compliance with quality indicators, assessment of mortality risk, and correlation of parenteral treatment strategies with clinical outcomes. More than 100,000 patient episodes have been recorded in the registry. These patient data have been reviewed and reveal important findings regarding acute decompensated heart failure and potential opportunities to improve the quality of care. PMID- 16036036 TI - The emerging role of natriuretic peptides in the diagnosis and treatment of decompensated heart failure. AB - B-type natriuretic peptide (BNP) is a cardiac neurohormone and is released as prepro BNP and then enzymatically cleaved to the N-terminal proBNP (NT-proBNP) and BNP upon ventricular myocyte stretch. Blood measurements of BNP and NT-proBNP have been used to identify patients with heart failure (HF). Clinical considerations for these tests include their half-lives in plasma, dependence on renal function for clearance, interpretation of their units of measure, and the rapid availability of the test results. The BNP assay is currently used as a diagnostic and prognostic aid in HF. In general, a BNP level below 100 pg/mL excludes acutely decompensated HF. Recombinant, human BNP (nesiritide) is an approved intravenous treatment for acute, decompensated HF. This paper reviews the literature concerning the use of this peptide as a diagnostic test and as an intravenous therapy. PMID- 16036037 TI - Levosimendan: a calcium-sensitizing agent for the treatment of patients with decompensated heart failure. AB - Levosimendan is a new inodilator. Its mechanism of action includes calcium sensitization of contractile proteins and the opening of adenosine triphosphate dependent K channels. The combination of positive inotropy with anti-ischemic effects of K-channel opening offers potential benefits in comparison with currently available intravenous inotropes, which are contraindicated in patients with ongoing myocardial ischemia. Levosimendan has been extensively studied in various animal models of heart failure, in which the drug has increased contractility without adverse effects on diastolic function. These results have been repeated in patients with heart failure, in whom levosimendan dose dependently increases cardiac output and reduces pulmonary capillary wedge pressure. The active metabolite of levosimendan (OR-1896) significantly prolongs the duration of the hemodynamic effects of the therapeutic 24-hour levosimendan infusion. Levosimendan has been studied in two major trials with decompensated patients (LIDO and RUSSLAN), in which it showed outcome benefits in comparison with dobutamine and placebo, respectively. A third comparative study (CASINO) recently suggested mortality benefits with levosimendan over placebo and dobutamine. Currently, two large prospective trials (SURVIVE and REVIVE) in patients who are hospitalized because of worsening heart failure are underway. These trials will conclusively prove whether levosimendan should be added to the standard treatment in patients who are hospitalized because of cardiac decompensation. PMID- 16036038 TI - Atrial fibrillation post-myocardial infarction: frequency, consequences, and management. AB - Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia and is a frequent complication of acute myocardial infarction (MI). AF occurs in 5% to 10% of patients who have received fibrinolysis. Post-MI AF is more common in older patients, in patients with heart failure, and after more extensive infarction. Postinfarction prognosis is worse among patients complicated by AF--indeed, mortality and morbid events including stroke, thromboembolism, and heart failure are increased in this group. It is unclear as to whether AF directly reduces survival or merely demarcates patients at higher overall risk. Despite its frequent occurrence and deleterious influence on outcomes, randomized data regarding management of AF after acute MI are scarce. This review summarizes recent data charting the incidence of AF after acute MI and describes features associated with its occurrence. Clinical sequelae and current principles in treatment are also discussed. PMID- 16036039 TI - Do statins prevent heart failure in patients after myocardial infarction? AB - 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, or statins, reduce morbidity and mortality in patients with coronary artery disease (CAD). Because CAD is the major cause of heart failure (HF) in developed countries, prevention of CAD may result in reduced HF. Evidence from randomized trials on lipid reduction (Cholesterol and Recurrent Events and the Scandinavian Simvastatin Survival Study) has shown statins to decrease progression to HF. Recently, many beneficial effects of statins have been demonstrated beyond cholesterol lowering. These agents improve endothelial function, exhibit anti-inflammatory properties, and prevent cardiac hypertrophy. Experimental studies have shown attenuation of left ventricular remodeling after myocardial infarction, possibly through reduced oxidative stress. However, no clinical evidence exists to support an effect on ventricular remodeling. Small, short-lasting clinical studies have also suggested that statin therapy might be associated with improved survival in ischemic and nonischemic HF. PMID- 16036040 TI - Current concepts for the neurohormonal management of left ventricular dysfunction after myocardial infarction. AB - The importance of addressing neurohormonal activation in patients after a myocardial infarction is now well-appreciated. Inhibition of the renin angiotensin-aldosterone axis and the sympathetic nervous system can result in improved cardiac function and survival. As we learn more about other systems, we should be able to realize further benefits. In particular, the roles of endothelin, matrix metalloproteinases, and cytokines in remodeling are being investigated, with the potential to result in better outcomes for patients. PMID- 16036041 TI - Aldosterone in heart failure: pathophysiology and treatment. AB - Aldosterone produces adverse effects on the vasculature (endothelial dysfunction) and on the myocardium (myocardial fibrosis). These effects have adverse clinical consequences that result in increases in deaths caused by sudden death and by progressive heart failure. The Randomized Aldactone Evaluation Study and the Eplerenone Post-acute Myocardial Infarction Heart Failure Efficacy and Survival Study show this clearly. Aldosterone blockade should become a regular third neuroendocrine-blocking drug in patients with chronic heart failure. PMID- 16036042 TI - Anemia and heart failure. AB - Over the past few years, anemia has emerged as a powerful independent predictor of adverse outcomes in chronic heart failure (CHF). It affects up to 50% of patients with CHF, depending on the definition of anemia used and on the population studied. Even small reductions in hemoglobin are associated with worse outcome. However, the causes of anemia in CHF remain unclear, although impairment of renal function and inflammatory cytokines are proposed mechanisms. Both may act through impairment of the synthesis or action of erythropoietin. Preliminary studies have demonstrated improvement in symptoms, exercise tolerance, quality of life, and reductions in hospitalizations when patients with severe CHF were treated with erythropoietin. The benefits and the potential risks of such therapies will be further addressed in upcoming larger randomized trials. The recent interest in anemia reflects a new perspective in heart failure therapy, focusing on non-cardiovascular comorbidities. PMID- 16036043 TI - Type 5 phosphodiesterase inhibition in heart failure and pulmonary hypertension. AB - The availability of selective inhibitors of the cyclic guanosine monophosphate (cGMP)-specific type 5 phosphodiesterase (PDE5) has created increasing interest in unlocking the therapeutic potential of PDE5 inhibition in cardiovascular diseases that are marked by dysfunction of nitric oxide (NO)-cGMP signaling. Pulmonary arterial hypertension (PAH) and heart failure (HF) are characterized by pulmonary arterial vasoconstriction that is thought to be caused by relative deficiencies of vasodilators such as NO and exaggerated production of vasoconstrictors such as endothelin. PDE5 is abundant in the pulmonary vasculature where it catabolizes cGMP, the second messenger of NO. Inhibition of PDE5 has been shown to lower pulmonary vascular resistance in PAH and HF by augmenting local cGMP. This review outlines the therapeutic potential of PDE5 inhibition for the treatment of PAH and HF. PMID- 16036044 TI - Vasopressin antagonists in heart failure. AB - The recrudescence of interest in manipulation of the arginine vasopressin system and especially of V2 vasopressin receptor blockade in heart failure stems from the limited efficacy and possible detrimental effects of loop diuretics. The "braking phenomenon," hypertrophy of the collecting duct cells, and altered pharmacodynamics contribute to loop diuretic resistance in heart failure. Selective (tolvaptan) and nonselective (conivaptan) V2 vasopressin receptor antagonists now known as "vaptans" promote free-water excretion that is labeled aquaresis and correct hyponatremia in patients with severe heart failure. A large mortality study with tolvaptan in heart failure is presently ongoing. PMID- 16036045 TI - Molecular remodeling in the failing human heart. AB - Heart failure is a progressive and fatal disease process. Arrhythmias and progressive cardiac dysfunction account for most of the morbidity and mortality in patients with heart failure. In general, the physiologic mechanisms responsible for progressive myocyte dysfunction, remodeling, and arrhythmias involve signaling mechanisms that alter myocardial gene expression. These changes in gene expression are complex and involve contractile proteins, ion channels, Ca++ handling, apoptosis, cell metabolism, the extracellular matrix, signal transduction pathways, and growth factors. PMID- 16036046 TI - Disruptions and detours in the myocardial matrix highway and heart failure. AB - Myocardial remodeling invariably occurs in congestive heart failure (CHF) and is a response to a prolonged cardiovascular stress, which is characterized by a cascade of compensatory structural events. Remodeling of the myocardial interstitium occurs in CHF and likely contributes to the progression of the remodeling process. The myocardial matrix can be considered a biological highway in which a large amount of signaling proteins and structural proteins are being moved within the interstitium, entering and exiting the interstitial space, and docking to cellular components. The rates at which these events occur can accelerate and decelerate depending on the particular cardiac disease state and thereby can alter the course of myocardial remodeling. Once considered merely a scaffolding to align cells, the matrix plays a complex and divergent role in influencing cell behavior. For example, the matrix has a functional role in cell migration, proliferation, adhesion, and cell-to-cell signaling. In light of this, the myocardial matrix should not be regarded as merely a static structure, but rather, as a complex system of dynamic interactions between matrix molecules, signaling proteins, and transmembrane proteins. Specific strategies that are targeted at modifying activity along this matrix highway will likely alter the course of myocardial remodeling and heart failure. PMID- 16036047 TI - Role of apoptosis in ventricular remodeling. AB - Cell loss is a common feature in the failing heart, and this contributes to the relentless progression seen in patients with heart failure. Apoptosis is one of the most common causes of cell loss in animals and humans with heart failure. There is increasing evidence that apoptosis, even while occurring in a low-grade manner, can mediate heart failure. Moreover, inhibiting apoptosis successfully prevents or attenuates heart failure in several animal models. More importantly, apoptosis is one of the few mechanisms that can be easily modulated using pharmacologic or gene therapy approaches. Animal data, obtained in the past few years, have proven the feasibility and success of this approach toward altering the natural history of heart failure. Human studies are pending, but a number of issues such as the type of inhibitor and its optimum timing/dose will need to be resolved before this becomes a reality. Nevertheless, these data will accrue over time, and anti-apoptotic therapy is likely to emerge as an important form of heart failure therapy. PMID- 16036048 TI - Marathoners or couch potatoes: what is the role of exercise in the management of heart failure? AB - Patients with chronic heart failure have diminished exercise capacity as a major aspect of their clinical syndrome, regardless of the cause of their left ventricular contractile dysfunction. The mechanisms for the reduction in exercise capacity are multifactorial and include central cardiac, peripheral vascular, respiratory, and skeletal muscle maladaptations that accompany the pathophysiology of heart failure. Increased sympathetic nervous system activity and elevations in circulating neurohormones and cytokines also influence the cardiovascular and metabolic responses to exercise in these patients. Despite the improvements in clinical outcomes with beta-blockers and resynchronization therapy in heart failure patients, their exercise capacity remains substantially reduced when compared with normal age-matched patients. Exercise training has the potential to reverse or improve most of the abnormal physiologic responses to exercise in these patients, and it may serve as adjunctive therapy to standard medical care of these patients. In addition, a body of evidence is being accumulated that suggests that exercise training itself has important secondary prevention benefit in these patients. This review identifies the potential mechanisms whereby exercise training may improve exercise capacity in patients with chronic heart failure and presents the current information regarding clinical outcomes of this therapy. PMID- 16036049 TI - Pacing and heart failure: should all patients receive a biventricular device? AB - The increasing adoption of biventricular pacing as a means to achieve interventricular, intraventricular, and atrioventricular synchronization affords clinicians a powerful nonpharmacologic tool in the treatment of congestive heart failure. Current guidelines support the use of cardiac resynchronization therapy (CRT) in patients with the combination of severe systolic ventricular dysfunction, symptomatic heart failure, and evidence of conduction delay on the surface electrocardiogram. Current research aims include investigation of differing clinical parameters and imaging techniques that will reliably differentiate patients who may or may not benefit from CRT. Furthermore, refinements in the techniques of lead placement and intrachamber timing may further improve the proportion of patients who achieve a favorable response from CRT. This article briefly reviews the history of CRT and summarizes some of the current trends and data relating to CRT. PMID- 16036050 TI - The role of prophylactic implantable cardioverter defibrillators in heart failure: recent trials usher in a new era of device therapy. AB - Sudden cardiac death (SCD) manifested as ventricular fibrillation or sustained ventricular tachycardia has been a major focus of cardiovascular research for more than three decades. Although mortality in patients with heart failure (HF) caused by left ventricular systolic dysfunction has declined in recent years through effective pharmacotherapeutic strategies, SCD remains the major cause of death in symptomatic HF, with little improvement by drug therapy. Although it is clear that the implantable cardioverter defibrillator (ICD) is efficacious and should be used to prevent a recurrence of sustained ventricular arrhythmia (secondary prevention) in most patients, the guidelines for prophylactic use of ICDs (primary prevention) are less well defined. The results of recent clinical trials examining the efficacy of prophylactic ICD therapy in HF patients have clarified the role of ICD treatment in this population. This article reviews these trials and summarizes our current approach to the prevention of SCD in HF. PMID- 16036051 TI - A heart failure specialist's perspective on cardiac surgery for heart failure. AB - Aside from cardiac transplantation, ventricular assist devices, and the total artificial heart, cardiac surgery now also plays a major role in the overall management of the heart failure patient. For patients with heart failure, cardiac surgery has steadily moved from being a predominant rescue procedure (eg, aneursymectomy, rupture repair, transplantation) to surgical interventions that can prevent or delay the progression of cardiac dysfunction and failure; these operations now include coronary artery bypass surgery, ventricular restoration, and valvular repair/replacement. This article discusses the role and impact of these specific surgical interventions in the setting of ventricular dysfunction and heart failure. PMID- 16036052 TI - The addition of angiotensin receptor blockers to angiotensin-converting enzyme inhibitors--what has time told us? AB - The neurohormonal hypothesis for the pathogenesis of heart failure found an early champion in the angiotensin-converting enzyme (ACE) inhibitors. More recently, the beta-blockers and aldosterone receptor antagonists have provided significant support by demonstrating marked additive clinical benefit. Within this context, angiotensin receptor blockers (ARBs) were specifically designed to antagonize one of the most potent contributors to the development and progression of heart failure, angiotensin. This review discusses the recent evidence for the addition of ARBs to standard therapy with ACE inhibitors and suggests how this evidence may be used to care for patients. PMID- 16036053 TI - The risks and benefits of aldosterone antagonists. AB - Spironolactone and eplerenone are mineralocorticoid- blocking agents used for their ability to block a host of epithelial and nonepithelial actions of aldosterone. These compounds are of proven benefit in reducing blood pressure and urine protein excretion, and in conferring cardiovascular gain in diverse circumstances of heart failure. However, as enthusiasm grows for use of mineralocorticoid-blocking agents, the risks inherent to use of such drugs become more pertinent. Whereas the endocrine side effects of spironolactone are in reality little more than a cosmetic disfigurement, the potassium-sparing properties of spironolactone and eplerenone can prove life-threatening if hyperkalemia develops. However, for most patients the risk of developing hyperkalemia should not dissuade the prudent clinician from use of these compounds. Hyperkalemia should be considered as a possibility in any patient receiving these medications and as such is best addressed preemptively. PMID- 16036054 TI - The impact of race on response to RAAS inhibition. AB - Retrospective analyses of the Studies of Left Ventricular Dysfunction (SOLVD) and Vasodilator Heart Failure Trials (V-HeFT) have addressed the question of whether angiotensin-converting enzyme (ACE) inhibitors are equally efficacious in black patients and white patients with heart failure. In SOLVD, there was no ethnic difference in the efficacy of enalapril for reducing mortality and preventing the development of heart failure, but enalapril was more effective in whites in reducing hospitalizations. In V-HeFT II, enalapril was more efficacious than the combination of isosorbide dinitrate and hydralazine in whites in reducing mortality, but not in blacks. However, the combination of isosorbide dinitrate and hydralazine may be particularly advantageous in black patients as suggested by V-HeFT I and the recent African American Heart Failure Trial. In aggregate, the available data suggest that ACE inhibitors should remain a cornerstone of therapy for heart failure with a reduced ejection fraction in white patients and black patients. PMID- 16036055 TI - Imaging the renin-angiotensin- aldosterone system in the heart. AB - The influence of the renin-angiotensin system (RAS) is recognized in cardiac and vascular injury. An extrinsic RAS has been known for decades, and an equally important intrinsic RAS has been discovered recently. The latter leads to pathologic tissue alterations in the absence of systemic stimuli and may be the main source of local tissue effects of RAS. A new radiotracer fluorobenzoyl lisinopril was synthesized by radiolabeling benzoic acid active ester with 18F and reacting that with the epsilon-amino group of lisinopril. The presence of angiotensin-converting enzyme (ACE) activity and angiotensin II receptors was examined in relation to myocardial fibrosis. This tissue-specific radioligand represents the first study of ACE in the human heart. This article presents preliminary data on imaging the RAS in the human cardiac tissue and discusses the potential for clinical application of these imaging techniques to human patients. PMID- 16036056 TI - Combination pharmacologic therapies for heart failure: what next after angiotensin-converting enzyme inhibitors and beta-blockers? AB - Although the introduction of angiotensin-converting enzyme (ACE) inhibitors and beta-adrenergic blockers has resulted in significant improvements in the management of heart failure (HF), morbidity and mortality remain high. Therefore, additional approaches have been sought to discover newer agents that might add incremental benefit. Although not all of these approaches have been successful, there have been some notable new approaches to therapy that have shown benefit or may be promising in terms of additional benefit. Most of these agents are targeted to achieve a more global neurohormonal blockade aiming to reduce or potentially reverse the ventricular remodeling process that occurs in HF. Some of the newer approaches aim for targets other than neurohormonal systems, eg, effects on myocardial metabolism or the vasculature. This article reviews the latest advances in pharmacologic therapy in HF, looking at several trials that may have a significant impact on the treatment of HF. We also discuss several newer agents with promising potential in HF management. PMID- 16036057 TI - When to initiate beta-blockers in heart failure: is it ever too early? AB - Overwhelming clinical trial evidence confirms the efficacy and safety of beta blockers in patients with heart failure (HF) caused by systolic dysfunction. beta Blockers are recommended in national HF guidelines as standard of care therapy. Yet there is also a large body of evidence demonstrating that the use of beta blockers for HF is seriously inadequate under conventional care. This HF treatment gap is due, in part, to the persistence of perceptions--despite recent evidence to the contrary--that beta-blocker therapy should be delayed until HF patients have been titrated to target doses of angiotensin-converting enzyme inhibitors and have been stable for at least 2 to 4 weeks after hospital discharge, and that early beta-blocker initiation results in a substantial risk of worsening HF. Conversely, recent clinical trial evidence substantiates that beta-blockers significantly reduce the risk of mortality and morbidity, including hospitalization for worsening HF, and have produced early survival benefits in patients with HF. It has also become evident that in-hospital initiation of life prolonging cardiovascular therapies, including beta-blockers, has a positive impact on clinical outcomes and on long-term patient compliance. Overwhelming clinical evidence suggests that beta-blockers should be administered to all stable HF patients without contraindication and that this therapy should be initiated as soon as possible to ensure that patients derive early and long-term improvements in clinical outcomes. PMID- 16036058 TI - Beta-blockers in selected heart failure populations. AB - Despite the overwhelming evidence of the beneficial effect of beta-blockers in systolic heart failure, questions have been raised as to whether they provide similar benefits to women, the elderly, and blacks. Also, concerns about the use of beta-blockers in diabetes, chronic obstructive lung disease, and peripheral vascular disease (PVD) have limited their use in populations with these diseases. A review of the experience generated from the three beta-blocker survival heart failure trials confirms the extension of similar beneficial effects to women, the elderly, blacks, and diabetic patients. There is strong support in the literature for the use of selective beta-blockers in patients with mild to moderate chronic obstructive pulmonary disease and encouraging data on their use in patients with mild to moderate PVD. PMID- 16036059 TI - Osteocyte and bone structure. AB - The osteocyte is the most abundant cell type of bone. There are approximately 10 times as many osteocytes as osteoblasts in adult human bone, and the number of osteoclasts is only a fraction of the number of osteoblasts. Our current knowledge of the role of osteocytes in bone metabolism is far behind our insight into the properties and functions of the osteoblasts and osteoclasts. However, the striking structural design of bone predicts an important role for osteocytes in determining bone structure. Over the past several years, the role of osteocytes as the professional mechanosensory cells of bone, and the lacunocanalicular porosity as the structure that mediates mechanosensing have become clear. Strain-derived flow of interstitial fluid through this porosity seems to mechanically activate the osteocytes, as well as ensure transport of cell signaling molecules, nutrients, and waste products. This concept explains local bone gain and loss--as well as remodeling in response to fatigue damage--as processes supervised by mechanosensitive osteocytes. Alignment during remodeling seems to occur as a result of the osteocyte's sensing different canalicular flow patterns around the cutting cone and reversal zone during loading, therefore determining the bone's structure. PMID- 16036060 TI - Combining high-resolution micro-computed tomography with material composition to define the quality of bone tissue. AB - Atraumatic fractures of the skeleton in osteoporotic patients are directly related to a deterioration of bone strength. However, the failure of the bone tissue to withstand functional load bearing cannot be explained as a simple decrease in bone mineral density (quantity); strength is also significantly dependent upon bone quality. While a formal definition of bone quality is somewhat elusive, at the very least, it incorporates architectural, physical, and biologic factors that are critical to bone strength. Such factors include bone morphology (ie, trabecular connectivity, cross-sectional geometry, longitudinal curvature); the tissue's material properties (eg, stiffness, strength); its chemical composition and architecture (eg, ratio of calcium to other components of the organic and/or inorganic phase, collagen orientation, porosity, permeability); and the viability of the tissue (eg, responsivity of the bone cell population). Combining high-resolution structural indices of bone, as determined by micro-computed tomography; material properties determined by nanoindentation; and the chemical make-up of bone, as determined by infrared spectroscopy, helps to provide critical information toward a more comprehensive assessment of the interdependence of bone quality, quantity, and fracture risk. PMID- 16036061 TI - Noninvasive assessment of bone structure. AB - Bone fragility is determined by bone mass and trabecular structure. While bone mass can be readily measured as bone density, bone trabecular structure cannot be easily assessed by currently available methods. The realization of the importance of bone structure in determining fracture risk has led to the development of several imaging modalities aimed at evaluating the contribution of bone quality to its biomechanical strength and fragility. High-resolution magnetic resonance imaging and computed tomography have limited spatial resolution and high cost but have a potential to generate true three-dimensional images of trabecular structure in vivo. Bone radiographs subjected to various forms of texture analysis have higher resolution and lower cost but provide only a two-dimensional representation of bone structure. Both two- and three-dimensional methods have been shown to predict biomechanical strength in vitro and to differentiate between subjects with and without fractures in vivo. Therefore, all of these methods deserve closer evaluation and also need further technical improvements before they can be considered for use in clinical practice. PMID- 16036062 TI - Hip geometry and its role in fracture: what do we know so far? AB - As the number of hip fractures continues to rise, finding better ways to identify people at risk becomes crucial. In the past decade, several measures of hip geometry have been studied as possible risk factors for hip fracture. Among them, hip axis length (HAL) shows the greatest promise for enhancing fracture risk assessment in the clinical setting, followed by neck shaft angle (NSA) and femoral neck width (FNW). Studies have shown that both age and/or a loss of body weight are associated with changes in some of the geometric parameters, which subsequently indicate the decrease in hip strength. The greater hip strength in black women and men resulting in a lower incidence of fractures compared with white women is also attributed to more favorable geometric parameters. Asian women, who have a lower incidence of fractures than white women, have a shorter HAL and a smaller NSA. In general, a longer HAL and a greater NSA and FNW all increase the risk of fracture, though controversies exist due to the use of different subject populations and measurement tools. Overall evidence suggests assessing hip geometry para-meters can significantly improve the ability of identifying people at risk of fracture, but more development in measurement software and more research are necessary to make it applicable in clinical settings. PMID- 16036063 TI - How to grow bone to treat osteoporosis and mend fractures. AB - The growing number of patients with osteoporosis in our aging population need "anabolic" drugs to stimulate bone growth, improve bone microarchitecture, and accelerate fracture healing. Potent anabolic agents such as parathyroid hormone (PTH) and some of its adenylyl cyclase-stimulating fragments are either on their way, or have just now reached the clinic. This article discusses how PTHs might stimulate bone growth. The controversial bone anabolic activities of the widely used cholesterol-lowering lipophilic statins and how they might stimulate bone growth are also probed. Also, evidence is presented for leptin, a controller of body fat stores and the ovarian cycle. It has the remarkable property of being an anabolic and antianabolic that uses a hypothalamic factor to restrain osteoblast activity but by itself stimulates osteoblasts and inhibits osteoclasts. PMID- 16036064 TI - Mechanism of action of bisphosphonates. AB - In recent years, substantial progress has been made in understanding the mechanism for bisphosphonate suppression of bone turnover. Bisphosphonates can now be distinguished based on their molecular and cellular mechanisms of action. Simple bisphosphonates such as clodronate and etidronate inhibit bone resorption through induction of osteoclast apoptosis. Clodronate, and perhaps etidronate, triggers apoptosis by generating a toxic analog of adenosine triphosphate, which then targets the mitochondria, the energy center within the cell. For nitrogen containing bisphosphonates, the direct intracellular target is the enzyme farnesyl diphosphate synthase in the cholesterol biosynthetic pathway. Its inhibition suppresses a process called protein geranylgeranylation, which is essential for the basic cellular processes required for osteoclastic bone resorption. Although nitrogen-containing bisphosphonates can induce osteoclast apoptosis, this is not necessary for their inhibition of bone resorption. PMID- 16036065 TI - Role of cytokines in postmenopausal bone loss. AB - It is increasingly recognized that changes in cytokine activity are involved in the pathophysiology of postmenopausal bone loss in many ways. Estrogen may affect skeletal cytokine activity by diverse mechanisms such as direct interference with cytokine gene transcription, regulation of major signaling molecules, changes in lymphocyte function, and central effects on the hypothalamic-pituitary-adrenal axis. This article summarizes our current knowledge of these interactions and discusses the possible implications with respect to the prevention and treatment of postmenopausal osteoporosis. PMID- 16036066 TI - Review of guidelines for testing and treatment of osteoporosis. AB - Our knowledge on diagnosis and treatment of osteoporosis has steadily increased during the past decade. Several guidelines on treatment of osteoporosis are now available. Although there is heterogeneity in these recommendations, there are several common suggestions found. Case finding is advocated in all; however, it is generally acknowledged that further research is necessary to evaluate the effectiveness of case-finding strategies. Dual-energy x-ray absorptiometry is considered the gold standard for diagnosis of osteoporosis. The use of the T score is different for diagnostic purposes and for treatment decisions. Other bone measurement techniques are proposed as risk evaluation or as alternatives when dual-energy x-ray absorptiometry is not available. Bone markers are not considered for evaluation in clinical practice. Treatment options include general measures on lifestyle, fall prevention, calcium and vitamin D supplements, hormone therapy, raloxifene, and bisphosphonates. Consistent recommendations are found mostly for raloxifene in the prevention of vertebral fractures and for alendronate and risedronate in the prevention of vertebral and nonvertebral fractures, including hip fractures. PMID- 16036067 TI - Burden of osteoporosis and fractures. AB - Osteoporosis currently affects up to one in three women and one in 12 men. In 1990, there were 1.6 million hip fractures per annum worldwide and this number is estimated to reach 6 million by 2050. This increase in the number of fractures is due to an increase in the number of elderly people in the population, improved survival, and an increase in the age-specific fracture rates of unknown etiology. The rising number of osteoporotic fractures and their associated morbidity will place a heavy burden on future health care resources. In the United States, the cost for the management of osteoporosis has been estimated at $17 billion. The majority of this cost is spent on the acute surgical and medical management following hip fracture, and the subsequent rehabilitation. Currently, only minimal costs are utilized for treatment and prevention of osteoporosis. Hopefully, however, an accurate assessment of the burden of osteoporosis on the individual and the health care system will enable the targeting of resources to tackle this growing problem. With an increasing number of effective pharmaceutical interventions, it is critical that these agents are targeted to those at greatest risk for future fracture. This will ultimately reduce the burden of osteoporosis in future years. PMID- 16036068 TI - Male osteoporosis: epidemiology and pathophysiology. AB - In men, osteoporosis is a prevalent problem that is under-recognized and undertreated. Men 50 years and older have a 13% lifetime risk for fracture. Hip and vertebral fractures are associated with significant functional impairment and increased mortality in men. The morbidity and mortality following a fracture is also greater in men than it is in women. By improving our knowledge on the pathophysiology of osteoporosis in men, better management strategies for this condition may be developed. In recent years, there has been greater awareness of the biomechanic factors that contribute to bone strength, which may explain some of the differences in fracture incidence between men and women. There is also growing evidence to support the key role of estrogens in maintaining bone health in older men, similar to women. This review highlights our current understandings on the epidemiology and pathophysiology of male osteoporosis and its related fractures, with particular focus on the determinants of bone strength and the role of sex hormones on bone metabolism in men. PMID- 16036069 TI - Hip fracture: risk factors and outcomes. AB - Hip fracture is an important public health and personal burden, and this burden is anticipated to increase over the next several decades. Although white women experience the greatest lifetime risk of hip fracture, risk extends to men and to nonwhite populations. Bone strength, risk of falling, and individual clinical characteristics combine to affect the risk of hip fracture. Nearly $9 billion were expended in 1995 in the United States for the management of hip fractures. Hip fracture has important sequelae, including loss of bone and muscle mass. Mortality is significantly increased after hip fracture, and functional recovery is limited to less than 50% of those who fracture. About 25% of patients reside in long-term care facilities for a year or more after fracture, and the impact of hip fracture on health-related quality of life is considerable and long lasting. PMID- 16036070 TI - Bone remodeling markers: assessment of fracture risk and fracture risk reduction. AB - There is an increasing repertoire of laboratory tests available for assessing the bone remodeling process. Biochemical markers of bone remodeling can be measured in serum or urine, and have a number of potential roles in the management of fracture risk. Differences in remodeling between individuals might be related to fracture risk and could be used to target therapy. Change in remodeling with therapy could be related to fracture risk reduction and the choice of therapy could be influenced by knowledge of bone remodeling. Biochemical therapeutic monitoring may improve patient understanding and therapeutic adherence. The rate of bone remodeling is weakly predictive of fracture risk in individuals who are not receiving therapy; however, it is not clear whether this is independent of other risk factors. There is increasing evidence that change in bone mineral density does not explain fracture risk reduction with antiresorptive therapies, and that therapeutic benefit might be explained by change in bone turnover. Additional studies and information are required to allow these scientific advances to be translated into cost-effective and validated clinical protocols. Optimizing the precision and accuracy of bone turnover assessment remains an important priority. PMID- 16036071 TI - Quantitative ultrasound: use in the detection of fractures and in the assessment of bone composition. AB - Quantitative ultrasound (QUS) techniques have found widespread clinical use, but their specific role in clinical practice needs further refinement. This review discusses the ability of QUS approaches to predict the risk for prevalent vertebral fractures and the risk for future fractures. QUS approaches perform as well as central dual x-ray absorptiometry devices but with some disadvantages (at least for older QUS approaches) with regard to the predictive power for hip fractures. Technologic diversity of QUS approaches may lead to differences in performance. QUS also has the potential for assessing bone mineral density independent aspects of bone composition that are relevant for bone strength. For measurements at the calcaneus, it is evident that bone microstructure is the key determinant of QUS variables obtained. However, in most cases, microstructure is so highly correlated with bone mineral density that no separate assessment can be performed in clinical practice. At cortical sites, a selective assessment of bone properties is easier. Technologies need to be adapted to this purpose because requirements differ significantly from those desired for optimum fracture risk assessment. More importantly, multiple partially independent QUS variables need to be defined to assess complementary aspects of bone tissue. PMID- 16036072 TI - Current technologies in the evaluation of bone architecture. AB - In the context of osteoporosis, bone quality (which encompasses trabecular bone and cortical bone architecture), bone mineralization, turnover, and microdamage are all important, as are bone density and total content. Noninvasive assessment of bone quality has recently received considerable attention because bone density alone is not a surrogate for fracture prevalence and occurrence, and does not completely explain the therapeutic efficacy of emerging treatments. This paper focuses on the assessment of trabecular bone architecture, one of the factors that governs bone strength and may be categorized as a contributor to bone quality. The methodologies described include micro-computed tomography, magnetic resonance imaging, and computerized analysis of radiographic patterns of trabecular bone. PMID- 16036073 TI - Management of vitamin d depletion in postmenopausal women. AB - In the United States, the impact regarding the routine fortification of milk and other dairy products has been less than satisfactory in preventing vitamin D depletion in adults. As more individuals are being evaluated and treated for osteoporosis, it has become necessary to exclude vitamin D depletion as a cause, because of the likelihood that differences in vitamin D and calcium nutrition may modify the individual response to therapy. Increased prevalence of subclinical vitamin D depletion among vertebral and hip fracture patients and the beneficial effect of vitamin D therapy and calcium supplementation in preventing such fractures lend credence to this assumption. The prevalence of vitamin D depletion is increasing again, especially among older adults, precisely the group at greatest risk for osteoporotic fractures. PMID- 16036074 TI - Use of highly potent bisphosphonates in the treatment of osteoporosis. AB - Bisphosphonates are effective inhibitors of bone remodeling. In the clinical setting, these agents prevent bone loss, preserve bone architecture, and improve bone strength. Clinically significant reduction in the risk of spine and nonspine fractures is observed in patients known to be at risk for fracture. When administered appropriately, these drugs are well tolerated and have an excellent safety profile. Potent bisphosphonates are now the preferred treatment option to reduce the fracture risk in men and women with involutional and glucocorticoid induced osteoporosis. PMID- 16036075 TI - Skeletal and nonskeletal effects of raloxifene. AB - Raloxifene, a selective estrogen receptor modulator, is approved for the prevention and treatment of postmenopausal osteoporosis. Prevention studies with raloxifene have demonstrated preservation of bone density and suppression of bone turnover markers in young postmenopausal women. The Multiple Outcomes of Raloxifene Evaluation study was the pivotal treatment trial for raloxifene. It demonstrated significant reduction in the risk for vertebral fractures after 1 and 3 years. Significant reduction of nonvertebral fractures with raloxifene has not yet been demonstrated. In addition to the effects of raloxifene on bone, potentially beneficial effects on the cardiovascular system, breast, and uterus have been described. Most of these nonskeletal effects have been reported as secondary endpoints from large osteoporosis trials with raloxifene. Prospective, randomized, double-blind studies of raloxifene with breast cancer prevention and cardiovascular protection as primary endpoints are now underway. PMID- 16036076 TI - Therapeutic potential of parathyroid hormone. AB - Teriparatide, recombinant human parathyroid hormone (1-34) (rhPTH [1-34]), is approved for the treatment of osteoporosis in men and postmenopausal women at high risk for fracture. The best candidates are those who have already had vertebral compression fractures (symptomatic or asymptomatic) or other osteoporosis-related fractures, or those who have very low bone mineral density, in the T score range of -3.5 or below. Teriparatide is the first anabolic drug approved by the US Food and Drug Administration for osteoporosis. It not only dramatically improves bone mass, but also restores bone microarchitecture and increases bone diameter. All of these mechanisms contribute to increasing bone strength and reducing the risk for osteoporosis-related fractures. Although PTH has been used in combination with other agents such as estrogens, calcitonin, and bisphosphonates, the relative benefit of the combined approach versus teriparatide alone for fracture risk reduction has not been shown. In fact, some data suggest that initiating PTH and alendronate together in previously untreated patients or pretreating patients for a short time with alendronate before initiating PTH may somewhat reduce the anabolic response to PTH. There are many unanswered questions regarding PTH, such as the optimal duration of treatment, the optimal sequence of medications for severe osteoporosis, the mechanism of resistance to effect after 18 to 24 months, the effect of subsequent rechallenge with PTH and, most importantly, surrogates to measure PTH effect. PMID- 16036077 TI - Hormone replacement therapy: review, update, and remaining questions after the Women's Health Initiative Study. AB - Hormone replacement therapy (HRT) is a collective term to include a variety of sex steroids, estrogens, and progestogens, given to postmenopausal women at various doses and administered through various routes. HRT is used for the relief of menopausal symptoms, with many women experiencing considerable improvement in their quality of life. It is also used for the prevention and treatment of osteoporosis, which is as effective as other available therapies. Other potential benefits of HRT are the prevention of cardiovascular disease and dementia. The main risks of HRT are an increased incidence of breast cancer and a transient increase in venous thromboembolism. The Women's Health Initiative is the largest randomized clinical trial of HRT to date, and has demonstrated benefits and risks of one specific HRT regimen. Skeletal benefits have been confirmed in terms of fracture reduction, as has a small risk of breast cancer development. Further studies of other HRT regimens are urgently required to clarify the cardiovascular effects. PMID- 16036078 TI - Rational use of oral bisphosphonates for the treatment of osteoporosis. AB - Osteoporosis has become a major public health concern worldwide. Significant morbidity, mortality, and health expenditures are associated with osteoporotic fractures. Evidence from randomized controlled trials and meta- analyses supports the efficacy and safety of oral bisphosphonates as first-line pharmacologic agents for the prevention and treatment of osteoporosis. This article reviews the evidence demonstrating the beneficial effects of etidronate, alendronate, and risedronate on improving bone mass and preventing fractures in individuals with or at risk for osteoporosis. Issues surrounding dosing intervals and optimal duration of therapy are also discussed. We conclude that the nitrogen-containing bisphosphonates alendronate and risedronate are safe and efficacious agents in preventing and treating osteoporosis. They are superior to cyclical etidronate in improving appendicular bone mass, and in reducing future risk for nonvertebral fractures. Once-weekly dosing options with alendronate and risedronate are effective and reduce serious adverse drug effects, and therefore, are welcome additions to our therapeutic armamentarium. PMID- 16036079 TI - Intravenous bisphosphonate therapy for osteoporosis: where do we stand? AB - The use of oral bisphosphonates, particularly members of the aminobisphosphonate subclass, is well established for the treatment of osteoporosis. In a number of clinical settings, intravenous administration appears to be advantageous. However, current dosing and efficacy data are limited while definitive, long-term trials with some of these agents are ongoing. In this article, we review the available information and discuss the use of these drugs on that basis. PMID- 16036080 TI - Is it ethical to use placebos in osteoporosis clinical trials? AB - Because there are now effective agents for treatment of osteoporosis, the question is being raised as to whether or not it is ethical to have placebo controlled trials of new agents. It is ethical for patients who are at low risk of serious or irreversible harm to participate in placebo-controlled trials as long as they provide informed consent. Morbidity, mortality, and future fracture risk correlate with the presence of previous fractures, the number of previous fractures, whether or not the fracture is recent, and whether or not the fracture is clinically recognized. Lower-risk subjects who may be allowed to participate in placebo-controlled trials include those with low bone density but without a previous vertebral fracture, those with a single vertebral deformity that was not clinically recognized, and those with a vertebral fracture more than 2 years before. Higher-risk subjects who do not tolerate proven drugs or who have not responded to proven drugs may also participate. Even though it may be ethical for selected subjects to participate in placebo-controlled trials of new therapies for osteoporosis, steps should be taken to minimize their exposure (eg, unbalanced randomization, integration of outcomes, and powering trials to actual events rather than a projected number over 3 years), and treating patients who fracture or who fail to respond. PMID- 16036081 TI - Bone imaging: traditional techniques and their interpretation. AB - Patients with osteoporosis have to be diagnosed at an early stage to prevent fractures, the worst complication of this disease. Currently, measurements of bone mineral density (BMD) are used most frequently in the diagnosis of osteoporosis. According to the World Health Organization, osteoporosis is defined on the basis of BMD measurements that are compared with those of a healthy, young, female population. The best established techniques to measure BMD are dual x-ray absorptiometry of the lumbar spine and proximal femur and quantitative computed tomography of the lumbar spine. Conventional radiographs are not suited to assess bone mass, but they are important in the diagnosis and differential diagnosis of osteoporotic fractures. Quantitative ultrasound and structure analysis, based on high-resolution magnetic resonance imaging and computed tomography, are newer techniques in the diagnosis of osteoporosis that also focus on the assessment of bone structure. PMID- 16036082 TI - Current diagnostic techniques in the evaluation of bone architecture. AB - Bone quality, which encompasses trabecular bone and cortical bone architecture, bone mineralization, turnover, and microdamage, is an essential component of bone strength. Therefore, bone quality, bone density, and total content are the most important parameters in the diagnosis of osteoporosis. Noninvasive assessment of bone quality has recently received considerable attention because bone density alone is not a surrogate for fracture prevalence and occurrence, and does not completely explain the therapeutic efficacy of emerging treatments. This paper will focus on the noninvasive assessment of trabecular bone architecture, one of the factors that governs bone strength and may be categorized as a contributor to bone quality. The methodologies described will include magnetic resonance imaging, clinical multislice spiral computed tomography, and micro-computed tomography, along with computerized analysis of radiographic patterns of trabecular bone. PMID- 16036083 TI - Bone fractal analysis. AB - Fractal analysis is a quantitative method used to evaluate complex anatomic findings in their elementary component. Its application to biologic images, particularly to cancellous bones, has been well practiced within the past few years. The aims of these applications are to assess changes in bone and the loss of spongious architecture, indicate bone fragility, and to show the increased risk for fracture in primary or secondary osteoporosis. The applications are very promising to help complete the studies that can define bone density (bone mineral density by dual energy x-ray absorptiometry or quantitative computed tomography), and also have the capacity to distinguish the patients with a high or low risk for fracture. Their extension to the clinical fields, to define a test for fracture risk, is still limited by difficult application to the medical quantitative imaging of bones, between correct application at superficial bones and unreliable application to deep bones. The future evolution and validity do not depend upon fractal methods but upon well-detailed imaging of the bones in clinical conditions. PMID- 16036084 TI - Pitfalls in bone mineral density measurements. AB - Bone mineral density (BMD) measurements have been the single greatest advancement for osteoporosis. BMD measurements have helped define a prefracture diagnosis of osteoporosis, predict fracture risk in postmenopausal women and elderly men, and monitor the course of disease processes that negatively affect bone or therapeutic agents that can improve bone strength. Despite the large amount of clinical, epidemiologic, and basic science data that has advanced our understanding of BMD performance and interpretation, many pitfalls in BMD performance and interpretation pervade the practice of bone densitometry. However, all of these pitfalls can be overcome. Proper quality control and clinical interpretation of BMD computer printout reports are paramount for correct diagnosis, risk assessment, and serial BMD measurements. Though BMD application(s) are a clinical tool that can and should be used by many different primary care and specialty physicians, the performances and interpretations are not simple processes. Proper education and training in the use of BMD technologies provides the means of achieving the great intent that BMD measurements are capable of providing. PMID- 16036085 TI - Sex and ethnic differences in bone architecture. AB - Technologic developments and applications such as dual energy x-ray absorptiometry, magnetic resonance imaging, and computed tomography have enabled researchers to assess bone quantity (ie, bone mineral density) and bone quality (ie, bone architecture), which are two important and independent contributions to bone strength. Recent studies on sex differences in bone architecture indicate that a number of biomechanical variables lead to increased bone strength in males compared with females. Ethnic differences in bone architecture are less clear cut, indicating a need to identify and test the social and biologic variables that race and ethnicity represent. New methods using magnetic resonance imaging technology may become important in creating efficient and reliable in vivo methods of assessing features of bone architecture that are relevant to fracture risk and contribute to the elucidation of sex and ethnic differences in osteoporosis. PMID- 16036086 TI - Growth, children, and fractures. AB - Fractures in childhood have long been considered an unavoidable consequence of growth. Studies in recent years have documented the epidemiology of these very common fractures and have also documented considerable variation by fracture type and from country to country. There have also been a number of studies aimed at identifying risk factors particularly for the most common distal forearm fracture. These studies have consistently associated bone mineral density with these fractures. Other possible risk factors include obesity, physical inactivity, sports, cola beverages, calcium intake, risk taking, and coordination. While prospective studies are required to confirm these risk factors, accumulating evidence now suggests that a substantial proportion of fractures in children are preventable. PMID- 16036087 TI - Genes and osteoporosis. AB - Genes play an important role in the development of osteoporosis. Twin and family studies have consistently shown that peak bone mass, ultrasound properties of bone, skeletal geometry, bone turnover, and fracture are heritable. Yet, as we report in this paper, few candidate genes have been implicated without ambiguity. Osteoporosis is thought to be a polygenic disorder, determined by multiple genes and environmental risk factors, each with small to modest effect on bone mass and fracture. Here we argue that future success in finding genes is only possible with improved study design and the use of more rigorous analytic approaches that are now becoming available. PMID- 16036088 TI - Estrogen, androgen, and the pathogenesis of bone fragility in women and men. AB - During growth, estrogen deficiency in females may produce increased bone size as a result of removal of inhibition of periosteal apposition, while failed endosteal apposition produces thin cortices and trabeculae in the smaller bone. In males, androgen deficiency produces reduced periosteal and endosteal apposition, reduced bone size, and cortical and trabecular thickness. At completion of longitudinal growth, advancing age is associated with emergence of a negative bone balance in each basic multicellular unit (BMU) because of reduced bone formation. Bone loss occurs, but slowly because the remodeling rate is slow. In midlife, in females, estrogen deficiency increases remodeling rate, increases the volume of bone resorbed, and decreases the volume of bone formed in each of the numerous BMUs remodeling bone on its endosteal (endocortical, trabecular, intracortical) surfaces so bone loss accelerates. In males, remodeling rate remains slow and is driven largely by reduced bone formation in the BMU. Hypogonadism in 20% to 30% of elderly men contributes to bone loss. In both sexes, calcium malabsorption and secondary hyperparathyroidism may partly be sex hormone dependent and contributes to cortical bone loss. Concurrent periosteal apposition partly offsets endosteal bone loss, but less so in women than in men. More women than men fracture because their smaller skeleton incurs greater architectural damage and adapts less by periosteal apposition. Sex hormone deficiency during growth and aging is pivotal in the pathogenesis of bone fragility. PMID- 16036089 TI - The elusive concept of bone quality. AB - This paper outlines information from recent publications that aid our understanding of bone quality in relation to osteoporosis. In practical terms, bone quality designates the properties of bone that contribute to strength but are not assessed by bone densitometry. While osteoporosis is still defined in terms of bone density, the limitations of this approach, long questioned, have become indisputable. In parallel, the results of treatment trials of antiresorptive agents demonstrate that bone density is a flawed surrogate for bone fragility and a weak indicator of antifracture efficacy. The case for emphasizing bone turnover in assessing fracture risk, has become increasingly strong, and a redefinition of osteoporosis on this basis may well occur. New technologies for studying bone microstructure and matrix composition, merging with sophisticated biomechanical assessments, are advancing our ideas regarding bone "damageability" and its effects over time. PMID- 16036090 TI - Postmenopausal osteoporosis. AB - The postmenopausal state is the most common underlying cause for osteoporosis and most of our understanding of the condition relates to this group. Recent evidence has suggested that hormone therapy may no longer have the pre-eminent place it once held but the same studies have also confirmed its antifracture efficacy. However, an increasing variety of alternative therapies are available for the postmenopausal woman. Furthermore, recent understanding of the mechanisms of bone loss after estrogen withdrawal, suggest that newer even better targeted therapies might be developed in the future. PMID- 16036091 TI - Vertebroplasty and kyphoplasty: rapid pain relief for vertebral compression fractures. AB - Vertebroplasty and kyphoplasty are minimally invasive procedures aimed at relieving symptoms associated with vertebral compression fractures. They have become accepted, highly effective, safe procedures, and are known for their high patient satisfaction rates. Although they are image-guided percutaneous spine therapies, differences do exist. They have each arisen from different backgrounds and are championed by different subspecialties and each involves slightly different techniques. In addition, a prospective, randomized trial comparing the two techniques has yet to be performed. This lack of objective data allows market forces and opinions to have a large influence on utilization and patient referral. PMID- 16036092 TI - Oral antiresorptive therapy. AB - Oral antiresorptive agents play a pivotal role in the management of osteoporosis. This paper discusses the effects and potential future role of newer agents such as ibandronate. Alternative dosing schedules and routes of administration have become available and may improve fracture protection, compliance, and tolerability for the long term treatment of a chronic condition such as osteoporosis. Increasingly these agents are being used to reduce bone loss in other diseases associated with high risk for osteoporosis such as organ transplantation and cystic fibrosis. Such studies may act as prototypes for the extended use of this class of drugs in other chronic inflammatory disease states. The innovative, yet disappointing results from combining an antiresorptive agent (alendronate) with the anabolic effects of teriparatide is also discussed. The major problem that remains is the lack of direct comparison between the agents in terms of fracture endpoints. PMID- 16036093 TI - Combination/sequential therapy in osteoporosis. AB - Combination therapy includes the concomitant or sequential use of compounds sharing the same mode of action (eg, two or more inhibitors of bone resorption) or with distinct pathways of activity (eg, an inhibitor of resorption plus an anabolic agent). Combination use of antiresorptive agents may generate concerns, because of the risk of inducing oversuppression of bone turnover. However, if low doses of estrogen, used for the management of climacteric symptoms, are insufficient to normalize bone turnover, the addition of a bisphosphonate to hormone therapy may prove to be useful to achieve this objective. Patients pretreated with inhibitors of resorption, who have not achieved a full therapeutic response, are good candidates for treatment with anabolic agents. The increase in bone turnover that comes after the introduction of parathyroid hormone (PTH) in patients treated with an antiresorptive agent is similar to that observed in treatment-naive patients and the pattern of bone mineral density (BMD) increase is also identical, with the exception of a 6 month delay in the spine and hip BMD changes observed in prior alendronate-treated subjects. Current data discourage the concomitant use of alendronate and PTH since the bisphosphonate appears to blunt (in men and women) the anabolic action of PTH. Whether this applies to other bisphosphonates or inhibitors of resorption, remains unknown. The use of an inhibitor of bone resorption after completion of PTH treatment seems an appropriate way to maintain the skeletal benefits gained during therapy. Long-term clinical studies, using fractures as an endpoint should be initiated to better understand the clinical and pharmaco-economic interest of combination therapies in the management of osteoporosis. PMID- 16036094 TI - Falls, fractures, and hip pads. AB - Improvement of balance along with bone-enhancing pharmacotherapy can improve the level of an individual's physical activity and mobility. Balance can be improved with enhancement of postural proprioception and muscular strength. Postural deformities have been shown to impair quality of life of osteoporotic individuals. Kyphotic posture has been demonstrated to contribute to propensity to fall in osteoporotic individuals. Kyphotic posturing and gait disorders can be managed through proprioceptive training, use of a weighted kypho-orthosis, muscle re-education, and safe resistance exercises. Proprioceptive balance training can reduce falls and fracture. Sarcopenia and osteoporotic fractures create musculoskeletal challenges that cannot be met with pharmacotherapy alone. Bone loss, imbalance, and gait disorder along with cognitive concerns can increase with aging. Even in healthy persons, predisposition to falls increases with age related neuromuscular changes. Muscle strength decreases approximately 50% from age 30 to 80. Furthermore, the amount of body sway increases with reduction of proprioception. Therefore, measures that can decrease imbalance can reduce the risk for falls and fracture. In normal balance, ankle strategies are recruited rather than hip strategies. Strengthening of the lower extremity muscles reduces the risk for falls. Gait aids can also decrease the risk for falls. During a fall, the risk for hip fracture increases 30-fold if there is direct impact to the hip. The use of hip protectors can decrease the risk for hip fracture during a sideways fall. Training in effective safe-landing strategies should be included in fall prevention programs. PMID- 16036095 TI - Treatment of osteoporosis in chronic kidney disease and end-stage renal disease. AB - As glomerular filtration rate (GFR) declines from age-related bone loss or disease that specifically induces a decline in GFR, there are a number of metabolic bone conditions that may accompany the decline in GFR. These metabolic bone conditions span a spectrum from mild-to-severe secondary hyperparathyroidism in early stages of chronic kidney disease (CKD) to the development of additional heterogeneous forms of bone diseases each with its distinctly quantitative bone histomorphometric characteristics. Osteoporosis can also develop in patients with CKD and ESRD for many reasons beyond age-related bone loss and postmenopausal bone loss. The diagnosis of osteoporosis in patients with severe CKD or end-stage renal disease (ESRD) is not as easy to do as it is in patients with postmenopausal osteoporosis (PMO)--neither fragility fractures nor The World Health Organization bone mineral density criteria can be used to diagnose osteoporosis in this population since all forms of renal bone disease may fracture or have low "T scores". The diagnosis of osteoporosis in patients with CKD/ESRD must be done by first the exclusion of the other forms of renal osteodystrophy, by biochemical profiling or by double tetracycline-labeled bone biopsy; and the finding of low trabecular bone volume. In such patients, preliminary data would suggest that oral bisphosphonates seem to be safe and effective down to GFR levels of 15 mL/min. In patients with stage 5 CKD who are fracturing because of osteoporosis or who are on chronic glucocorticoids, reducing the oral bisphosphonate dosage to half of its usual prescribed dosing for PMO seems reasonable from known bisphosphonate pharmacokinetics, though we do need better scientific data to fully understand bisphosphonate usage in this population. PMID- 16036096 TI - Treatment of low bone mass in premenopausal women: when may it be appropriate? AB - The diagnosis and treatment of osteoporosis in postmenopausal women has been well defined. Criteria for making a diagnosis using bone density testing has been established and widely disseminated. However, the definition established by the World Health Organization for osteoporosis cannot be used for other populations such as premenopausal women without further study. Measuring bone mass in premenopausal women should be done infrequently and only if specific and significant risk factors are present. Premenopausal women have a low risk for fracturing but women with low bone mass are at a greater risk for developing fractures before and after menopause. The major reason for low bone mass is the failure to reach peak bone mass because of genetic influences, although some environ-mental factors may also play a role. Hormonal therapy for hypogonadism, treatment of diseases such as hyperpara-thyroidism, and keeping medications with detrimental effects on bone to a minimum are important components of treatment. Pharmacologic treatment of low bone mass in premenopausal women has not been well studied although on rare occasions it may be appropriate in women with low bone mass, defined as a Z score of -2 or less, and the presence of other diseases or risk factors. Caution is urged because of the unknown effects of treatment in this population. Further study of the implications of low bone mass and the effects of treatment in this group of women is needed. PMID- 16036097 TI - Does early PTH treatment compromise bone strength? The balance between remodeling, porosity, bone mineral, and bone size. AB - The treatment of osteoporotic women with recombinant human parathyroid hormone (rhPTH) increases bone mineral density and reduces fracture risk. However, there has been concern that the initiation of therapy in women with low bone mass may cause an early and transient increased fracture risk because PTH stimulates bone remodeling, which in its first phase is associated with bone resorption. Animal and human studies suggest, however, that the stimulation of remodeling caused by rhPTH(1-34) does not lead to a deterioration of bone's mechanical properties or to an increased fracture risk even early in the treatment. There are several reasons for this. Bone biomarkers associated with formation rise earlier than those associated with resorption, suggesting that there is an initial period prior to the stimulation of remodeling during which bone formation occurs on surfaces without prior resorption. This initial period of formation may protect the patient from the later small and transient losses that occur through remodeling. Moreover, the increased remodeling occurs on cancellous surfaces or close to the endosteal surface of bone, where its mechanical effect is minimal. Additionally, these transient losses may be compensated by periosteal apposition that maintains the overall strength of the bone. Thus, the early stimulation of bone formation without prior resorption, and the redistribution of bone from cancellous and endocortical surfaces to the periosteal surface combine to prevent the mechanical deterioration that could otherwise occur with a transient acceleration of bone remodeling in a patient with low bone mass. PMID- 16036098 TI - Glucocorticoid-induced osteoporosis: treatment options and guidelines. AB - Glucocorticoid-induced osteoporosis and fractures are the most frequent adverse effects of this class of medication. Recent advances in the pathophysiology, epidemiology, detection, and prevention of this complication of therapy provide hope for its amelioration in patients who require treatment with glucocorticoids. A number of effective pharmacologic agents are available and/or under study, and scientific organizations now provide guidelines for the prevention and treatment of glucocorticoid-induced osteoporosis. Unfortunately, consistent application of these guidelines remains suboptimal in many practice settings. PMID- 16036099 TI - Strontium ranelate: a new treatment for postmenopausal osteoporosis with a dual mode of action. AB - In vitro, strontium ranelate increases collagen and noncollagen protein synthesis by mature osteoblast-enriched cells. Its effects on bone formation were confirmed as the drug enhanced preosteoblastic cell replication. In the isolated osteoclast, preincubation of bone slices with strontium ranelate-induced dose dependent inhibition of the bone-resorbing activity of treated rat osteoclast. Strontium ranelate dose-dependently inhibited preosteoclast differentiation. Its effect in postmenopausal women with established osteoporosis was assessed during an international, prospective, double-blind, randomized, placebo-controlled phase 3 program comparing strontium ranelate 2 g daily with placebo. The 3-year analysis of the phase 3 study, Spinal Osteoporosis Therapeutic Intervention, evaluating the effect of strontium ranelate 2 g/day on vertebral fracture rates, revealed a significant 41% reduction in the relative risk of patients experiencing new vertebral fracture with strontium ranelate over 3 years. A second phase 3 study showed a significant reduction in the relative risk of experiencing a nonvertebral fracture in the group treated with strontium ranelate over 3 years. These results show that strontium ranelate is a new, effective, and safe treatment for vertebral and hip osteoporosis, with a unique mode of action, increasing bone formation and decreasing bone resorption leading to a rebalance of bone turnover in favor of bone formation. PMID- 16036100 TI - Microcracks in cortical bone: how do they affect bone biology? AB - Microcrack accumulation in cortical bone has been implicated in skeletal fragility and stress fractures. These cracks have also been shown to affect the mechanical and material properties of cortical bone. Their growth has been linked to osteocyte apoptosis and the initiation of the remodeling process, which also has a role in their repair. Clinically, osteoporosis is diagnosed using dual energy x-ray absorptiometry. However, evidence now indicates that bone mass alone is insufficient to satisfactorily explain the skeletal fragility of osteoporosis and consideration needs to be given to bone quality in the diagnosis and treatment of the disease. Bone quality includes parameters such as trabecular and cortical microarchitecture, morphology, bone turnover, degree of mineralization of the bone matrix, and significantly, the amount of microdamage present in the bone. Current clinical treatments concentrate on the inhibition of osteoclast activity to maintain bone mass in osteoporotic patients. However, these cells have a major role in removing existing microcracks from the bone matrix, and hence the use of bone resorption- inhibiting drugs may lead to insufficient bone repair and therefore an increase in microdamage accumulation and loss of bone quality. PMID- 16036101 TI - Regulation of bone mass by mechanical loading: microarchitecture and genetics. AB - For decades, the processes that couple bone architecture and mass to function have been investigated and characterized. It is well known, and now well accepted, that increases in exercise and loading of bone are associated with increased bone mass, and that disuse induces osteopenia. However, the mechanisms by which disuse leads to bone loss remain poorly understood, even in the 21st century. Clearly, the skeleton is able to perceive and respond to some general input(s) generated, or lost, as a consequence of mechanical unloading of bone that are distinct from habitual activity, so called functional adaptation. It is the focus of this paper to evaluate the evidence underlying roles for genetics, osteocytes, and interstitial fluid flow in mediating disuse osteopenia. PMID- 16036102 TI - Bone architecture and fracture. AB - Bones are designed to carry out their requirements effectively. One of these requirements is to resist fracture. Two other important requirements are to be stiff and to be light. Few theories of adaptive modeling distinguish modeling for adequate stiffness from modeling for adequate strength. Bones achieve their architecture partially through genetics, the rough form of the bone being laid down in the genes, and partially through response to normal loading. Normal loading rarely includes traumatic loading and bones are not usually well adapted to resist trauma, though they are probably well adapted to fatigue loading. Some aspects of architectural function, such as hollowness, are well understood. Some aspects, such as the need for uniform loading in impact, are less understood, and some, such as size effects, are only now beginning to be investigated. PMID- 16036103 TI - The relationship between bone mineral density and fracture risk. AB - Osteoporosis is a disorder characterized by low bone density and impaired bone strength which is an important risk factor for fracture in older adults. The diagnosis of osteoporosis in postmenopausal women is now based on bone density testing by dual energy x-ray absorptiometry but not other methodologies. However, a specific but arbitrary diagnostic threshold must be distinguished from a strategy to assess fracture risk. In untreated postmenopausal women and older men, bone density is an important, but not the only, determinant for fracture risk. Combining bone density measurements with other independent and validated risk factors for fracture provides a much more accurate assessment of an individual patient's risk for fracture than does bone density alone. The most important of these other risk factors are age and prior fracture history. Clinical guidelines will move away from recommending treatment at specific T scores toward intervention thresholds based on absolute fracture risk. By basing who to treat on fracture probability, therapy can be targeted to those patients who would receive the greatest benefit. PMID- 16036104 TI - Recent developments in trabecular bone characterization using ultrasound. AB - Currently available quantitative ultrasound technologies to assess cancellous bone are based on the measurements in transmission of speed of sound or slope of frequency-dependent attenuation (so called broadband ultrasonic attenuation). These two parameters are now considered as surrogate markers of site-matched bone mineral density. The ability of ultrasound techniques to provide non-bone mineral density-related bone properties (eg, microstructure) has not been clearly demonstrated yet. This is mainly because of two factors: a lack of understanding of ultrasound propagation with clear identification of the different underlying physical interactions; and the difficulty of performing experiments because of the limited sample size, the large number of statistical relationships to be tested with multiple variables, and the usual strong covariance observed between bone quantity and microarchitecture. The aim of this paper is to review the most recent development in the field of ultrasound characterization of trabecular bone. We present research work on ultrasound backscatter and how it could be used to estimate microarchitectural properties independently of bone quantity, and the first promising results obtained for the estimation of trabecular thickness. We then introduce numeric simulations of wave propagation through trabecular microarchitecture and show how it could contribute to elucidate and better characterize the physical underlying physics and result in more predictive models. These innovative acquisition schemes and the possibility of virtual experiments should altogether contribute to rapid advancement of ultrasonic bone characterization. PMID- 16036105 TI - Persistent STAT3 activation in colon cancer is associated with enhanced cell proliferation and tumor growth. AB - Colorectal carcinoma (CRC) is a major cause of morbidity and mortality in Western countries. It has so far been molecularly defined mainly by alterations of the Wnt pathway. We show here for the first time that aberrant activities of the signal transducer and activator of transcription STAT3 actively contribute to this malignancy and, thus, are a potential therapeutic target for CRC. Constitutive STAT3 activity was found to be abundant in dedifferentiated cancer cells and infiltrating lymphocytes of CRC samples, but not in non-neoplastic colon epithelium. Cell lines derived from malignant colorectal tumors lost persistent STAT3 activity in culture. However, implantation of colon carcinoma cells into nude mice resulted in restoration of STAT3 activity, suggesting a role of an extracellular stimulus within the tumor microenvironment as a trigger for STAT activation. STAT3 activity in CRC cells triggered through interleukin-6 or through a constitutively active STAT3 mutant promoted cancer cell multiplication, whereas STAT3 inhibition through a dominant-negative variant impaired IL-6-driven proliferation. Blockade of STAT3 activation in CRC-derived xenograft tumors slowed down their development, arguing for a contribution of STAT3 to colorectal tumor growth. PMID- 16036106 TI - Array-based comparative genomic hybridization identifies localized DNA amplifications and homozygous deletions in pancreatic cancer. AB - Pancreatic cancer, the fourth leading cause of cancer death in the United States, is frequently associated with the amplification and deletion of specific oncogenes and tumor-suppressor genes (TSGs), respectively. To identify such novel alterations and to discover the underlying genes, we performed comparative genomic hybridization on a set of 22 human pancreatic cancer cell lines, using cDNA microarrays measuring approximately 26,000 human genes (thereby providing an average mapping resolution of <60 kb). To define the subset of amplified and deleted genes with correspondingly altered expression, we also profiled mRNA levels in parallel using the same cDNA microarray platform. In total, we identified 14 high-level amplifications (38-4934 kb in size) and 15 homozygous deletions (46-725 kb). We discovered novel localized amplicons, suggesting previously unrecognized candidate oncogenes at 6p21, 7q21 (SMURF1, TRRAP), 11q22 (BIRC2, BIRC3), 12p12, 14q24 (TGFB3), 17q12, and 19q13. Likewise, we identified novel polymerase chain reaction-validated homozygous deletions indicating new candidate TSGs at 6q25, 8p23, 8p22 (TUSC3), 9q33 (TNC, TNFSF15), 10q22, 10q24 (CHUK), 11p15 (DKK3), 16q23, 18q23, 21q22 (PRDM15, ANKRD3), and Xp11. Our findings suggest candidate genes and pathways, which may contribute to the development or progression of pancreatic cancer. PMID- 16036107 TI - Cas IIgly induces apoptosis in glioma C6 cells in vitro and in vivo through caspase-dependent and caspase-independent mechanisms. AB - In this work, we investigated the effects of Casiopeina II-gly (Cas IIgly)--a new copper compound exhibiting antineoplastic activity--on glioma C6 cells under both in vitro and in vivo conditions, as an approach to identify potential therapeutic agents against malignant glioma. The exposure of C6 cells to Cas IIgly significantly inhibited cell proliferation, increased reactive oxygen species (ROS) formation, and induced apoptosis in a dose-dependent manner. In cultured C6 cells, Cas IIgly caused mitochondrio-nuclear translocation of apoptosis induction factor (AIF) and endonuclease G at all concentrations tested; in contrast, fragmentation of nucleosomal DNA, cytochrome c release, and caspase-3 activation were observed at high concentrations. Administration of N-acetyl-L-cystein, an antioxidant, resulted in significant inhibition of AIF translocation, nucleosomal DNA fragmentation, and caspase-3 activation induced by Cas IIgly. These results suggest that caspase-dependent and caspase-independent pathways both participate in apoptotic events elicited by Cas IIgly. ROS formation induced by Cas IIgly might also be involved in the mitochondrio-nuclear translocation of AIF and apoptosis. In addition, treatment of glioma C6-positive rats with Cas IIgly reduced tumor volume and mitotic and cell proliferation indexes, and increased apoptotic index. Our findings support the use of Cas IIgly for the treatment of malignant gliomas. PMID- 16036108 TI - Temporally regulated neural crest transcription factors distinguish neuroectodermal tumors of varying malignancy and differentiation. AB - Neuroectodermal tumor cells, like neural crest (NC) cells, are pluripotent, proliferative, and migratory. We tested the hypothesis that genetic programs essential to NC development are activated in neuroectodermal tumors. We examined the expression of transcription factors PAX3, PAX7, AP-2alpha, and SOX10 in human embryos and neuroectodermal tumors: neurofibroma, schwannoma, neuroblastoma, malignant nerve sheath tumor, melanoma, medulloblastoma, supratentorial primitive neuroectodermal tumor, and Ewing's sarcoma. We also examined the expression of P0, ERBB3, and STX, targets of SOX10, AP-2alpha, and PAX3, respectively. PAX3, AP 2alpha, and SOX10 were expressed sequentially in human NC development, whereas PAX7 was restricted to mesoderm. Tumors expressed PAX3, AP-2alpha, SOX10, and PAX7 in specific combinations. SOX10 and AP-2alpha were expressed in relatively differentiated neoplasms. The early NC marker, PAX3, and its homologue, PAX7, were detected in poorly differentiated tumors and tumors with malignant potential. Expression of NC transcription factors and target genes correlated. Transcription factors essential to NC development are thus present in neuroectodermal tumors. Correlation of specific NC transcription factors with phenotype, and with expression of specific downstream genes, provides evidence that these transcription factors actively influence gene expression and tumor behavior. These findings suggest that PAX3, PAX7, AP-2alpha, and SOX10 are potential markers of prognosis and targets for therapeutic intervention. PMID- 16036109 TI - The minimal set of genetic alterations required for conversion of primary human fibroblasts to cancer cells in the subrenal capsule assay. AB - Based on previous studies, a minimal set of genetic alterations that is required to convert normal human fibroblasts into cancer cells has been defined. Essential roles for telomere maintenance and alterations in phosphatase 2A activity were inferred from experiments in which tumorigenicity was tested by injecting cells under the skin of immunodeficient mice. However, in the present experiments, the combination of SV40 large T antigen and activated Ras, without hTERT or SV40 small t antigen, was sufficient to convert nine different primary human fibroblast cell strains to a fully malignant state. The malignant behavior of the cells was demonstrated by growth of the cells into invasive tumors when the cells were injected beneath the kidney capsule of immunodeficient mice. Lung metastases and circulating tumor cells were also detected. These tumors were not immortal; cells entered crisis, from which they could be rescued by expression of hTERT. However, the same cell populations were not tumorigenic when they were injected under the skin. In this site, tumorigenicity required the expression of hTERT and SV40 small t antigen as well as SV40 large T antigen and Ras. The cellular pathways targeted by SV40 large T antigen (p53 and pRb) and those targeted by activated Ras represent a minimal set of genetic alterations required for the conversion of normal human fibroblasts into cancer cells. PMID- 16036110 TI - Accelerated degradation of caspase-8 protein correlates with TRAIL resistance in a DLD1 human colon cancer cell line. AB - The tumor-selective cytotoxic effect of tumor necrosis factor-related apoptosis inducing ligand (TRAIL) makes TRAIL an attractive candidate as an anticancer agent. However, resistance to TRAIL poses a challenge in anticancer therapy with TRAIL. Therefore, characterizing the mechanisms of resistance and developing strategies to overcome the resistance are important steps toward successful TRAIL mediated cancer therapy. In this study, we investigated mechanisms of acquired TRAIL resistance in a colon cancer DLD1 cell line. Compared with the TRAIL susceptible DLD1 cell line, TRAIL-resistant DLD1/TRAIL-R cells have a low level of caspase-8 protein, but not its mRNA. Suppression of caspase-8 expression by siRNA in parental DLD1 cells led to TRAIL resistance. Restoration of caspase-8 protein expression by stable transfection rendered the DLD1/TRAIL-R cell line fully sensitive to TRAIL protein, suggesting that the low level of caspase-8 protein expression might be the culprit in TRAIL resistance in DLD1/TRAIL-R cells. Sequencing analysis of the caspase-8 coding region revealed a missense mutation that is present in both TRAIL-sensitive and TRAIL-resistant DLD1 cells. Subsequent study showed that the degradation of caspase-8 protein was accelerated in DLD1/TRAIL-R cells compared to parental DLD1 cells. Thus, accelerated degradation of caspase-8 protein is one of the mechanisms that lead to TRAIL resistance. PMID- 16036111 TI - High-resolution mapping of genomic imbalance and identification of gene expression profiles associated with differential chemotherapy response in serous epithelial ovarian cancer. AB - Array comparative genomic hybridization (aCGH) and microarray expression profiling were used to subclassify DNA and RNA alterations associated with differential response to chemotherapy in ovarian cancer. Two to 4 Mb interval arrays were used to map genomic imbalances in 26 sporadic serous ovarian tumors. Cytobands 1p36, 1q42-44, 6p22.1-p21.2, 7q32.1-q34 9q33.3-q34.3, 11p15.2, 13q12.2 q13.1, 13q21.31, 17q11.2, 17q24.2-q25.3, 18q12.2, and 21q21.2-q21.3 were found to be statistically associated with chemotherapy response, and novel regions of loss at 15q11.2-q15.1 and 17q21.32-q21.33 were identified. Gene expression profiles were obtained from a subset of these tumors and identified a group of genes whose differential expression was significantly associated with drug resistance. Within this group, five genes (GAPD, HMGB2, HSC70, GRP58, and HMGB1), previously shown to form a nuclear complex associated with resistance to DNA conformation-altering chemotherapeutic drugs in in vitro systems, may represent a novel class of genes associated with in vivo drug response in ovarian cancer patients. Although RNA expression change indicated only weak DNA copy number dependence, these data illustrate the value of molecular profiling at both the RNA and DNA levels to identify small genomic regions and gene subsets that could be associated with differential chemotherapy response in ovarian cancer. PMID- 16036112 TI - The hTERT and hTERC telomerase gene promoters are activated by the second exon of the adenoviral protein, E1A, identifying the transcriptional corepressor CtBP as a potential repressor of both genes. AB - Telomerase plays a role in the unlimited replicative capacity of the majority of cancer cells and provides a potential anticancer target. The regulation of telomerase is complex but transcriptional control of its two essential components, hTERC (RNA component) and hTERT (reverse transcriptase component), is of major importance. To investigate this further, we have used the adenoviral protein, E1A, as a tool to probe potential pathways involved in the control of telomerase transcription. The second exon of the adenoviral protein E1A activates both telomerase gene promoters in transient transfections. The corepressor, C terminal binding protein, is one of only two proteins known to bind to this region, and we propose that E1A activates both promoters by sequestering CtBP, thereby relieving repression. Activation by exon 2 of E1A involves the SP1 sites in both promoters, and consistent with this, SP1 and CtBP interact in coimmunoprecipitation studies. Modulation of the chromatin environment has been implicated in the regulation of hTERT transcription and appears to involve the SP1 sites. CtBP can be found within a histone-modifying complex and it is possible that a CtBP complex, associating with the SP1 sites, represses transcription from the telomerase promoters by modifying chromatin structure. PMID- 16036113 TI - Brain tumor tropism of transplanted human neural stem cells is induced by vascular endothelial growth factor. AB - The transplantation of neural stem cells (NSCs) offers a new potential therapeutic approach as a cell-based delivery system for gene therapy in brain tumors. This is based on the unique capacity of NSCs to migrate throughout the brain and to target invading tumor cells. However, the signals controlling the targeted migration of transplanted NSCs are poorly defined. We analyzed the in vitro and in vivo effects of angiogenic growth factors and protein extracts from surgical specimens of brain tumor patients on NSC migration. Here, we demonstrate that vascular endothelial growth factor (VEGF) is able to induce a long-range attraction of transplanted human NSCs from distant sites in the adult brain. Our results indicate that tumor-upregulated VEGF and angiogenic-activated microvasculature are relevant guidance signals for NSC tropism toward brain tumors. PMID- 16036114 TI - Parabens: a review of epidemiology, structure, allergenicity, and hormonal properties. AB - Parabens are the most commonly used preservatives in cosmetic, pharmaceutical, and industrial products. The epidemiology, structural properties, allergenicity, hormonal properties, and hypotheses regarding the "paraben paradox" are addressed in this review. PMID- 16036115 TI - Positive patch-test reactions to gold: patients' perception of relevance and the role of titanium dioxide in cosmetics. AB - BACKGROUND: Positive patch-test reactions to gold that are without obvious clinical relevance are common. We speculate that titanium dioxide in cosmetics and sunscreens may adsorb gold particles in jewelry that occasionally contacts facial skin and cause contact dermatitis on this area despite the absence of dermatitis under gold jewelry worn on the hands. OBJECTIVE: To identify subgroups of gold allergic patients who improve with avoidance of gold. METHODS: A questionnaire was mailed to 102 gold-allergic patients. RESULTS: There were 49 responses, of which all but one were from women. When asked to evaluate the statement that patch testing with gold was useful to them, one-third of respondents agreed completely, one-third agreed in part, and one-third disagreed. The majority of patients reported that their dermatitis improved after patch testing, but most were avoiding other allergens in addition to gold. CONCLUSIONS: Avoidance of gold earrings did not appear to benefit patients with earlobe dermatitis, but total avoidance of gold jewelry on the hands and wrists did seem to benefit a subgroup of patients with facial and eyelid dermatitis who wore powder, eye shadow, or foundation on affected areas. PMID- 16036116 TI - Use of consumer product ingredients for patch testing. AB - BACKGROUND: Patch testing for suspected sensitivity to cosmetics and other personal care products is usually done by testing with nonirritating products "as is" and by panels of antigens likely to contain causative ingredients. Most allergic reactions are reportedly due to sensitivity to either fragrances or preservatives. Although most preservatives found in patients' products are available for patch testing, only a small number of fragrance ingredients are available, and fragrance components are seldom labeled. Most personal care products contain many other ingredients, and unless the patient reacts to the whole product and the ingredients are obtained from the manufacturer, most of these are seldom tested. OBJECTIVE: To investigate the importance of testing with these other ingredients. METHODS: Investigators reviewed patch-test records of patients who presented with eruptions compatible with the use of their personal care products and who were tested with available ingredients that were listed on the labels of products they were using. This allowed testing with many ingredients in products that are too irritant for "as is" testing. Some of the results included those of persons who were tested in other series, so these were separated. RESULTS: Of patch tests with 52 cosmetic ingredients also tested in other series, 3.4% produced at least one+or greater reaction. Of those antigens tested only when present in products used by the patient, 55 of the 121 ingredients produced at least one+reaction, and about 3.6% of the test results were positive. CONCLUSIONS: Adding ingredients found in the patient's personal care products to patch tests done on those compatible with exposure increases the positive yield in patch testing, and the number of positive results is likely to increase as more ingredients are available for testing. PMID- 16036117 TI - Eczema herpeticum complicating Parthenium dermatitis. AB - Parthenium dermatitis is one of the most common causes of airborne contact dermatitis in India. Eczema herpeticum has been reported in association with various eczematous conditions, including Parthenium dermatitis. We report a case of eczema herpeticum in association with Parthenium dermatitis. Because Parthenium dermatitis is a common condition in this region, one should be aware of this complication so that appropriate treatment is not delayed. PMID- 16036118 TI - Allergic contact dermatitis from methyldibromoglutaronitrile. AB - BACKGROUND: Arriving at a diagnosis of allergic contact dermatitis is a multistep procedure including the establishing of contact allergy, demonstration of current exposure to the sensitizer, and assessment of clinical relevance. Sometimes, these steps are easy to get through; at other times, there may be problems with every step. OBJECTIVE: To demonstrate the possible difficulties and pitfalls in establishing the presence of contact allergy and diagnosing allergic contact dermatitis from exposure to the preservative methyldibromoglutaronitrile (MDBGN). METHODS: Simultaneous patch-testing with petrolatum preparations of MDBGN at various concentrations, use testing, and chemical analysis with high-performance liquid chromatography (HPLC). RESULTS: Contact allergy to MDBGN was established in two cases, with MDBGN in petrolatum at 0.5%. Results of HPLC investigation of moisturizers used by the patients and yielding positive results on patch and use tests disagreed with the information about preservatives on the labels of the moisturizers and with the Material Safety Data Sheets (MSDSs). CONCLUSIONS: Patch testing with MDBGN in petrolatum at a concentration of less than 0.5% may fail to diagnose a clinically relevant contact allergy. The information on labels of products, on MSDSs, and from manufacturers may not be reliable, which indicates the need for chemical analyses. PMID- 16036119 TI - Irritant contact dermatitis precipitating allergic contact dermatitis. PMID- 16036120 TI - Allergic contact dermatitis due to kojic acid. PMID- 16036121 TI - Delusions of grandeur, or what the public doesn't know about shortcuts. PMID- 16036122 TI - Implant placement. PMID- 16036123 TI - Managing congenitally missing lateral incisors. Part II: tooth-supported restorations. AB - Three treatment options exist for the replacement of congenitally missing lateral incisors: canine substitution, a tooth-supported restoration, or a single-tooth implant. Selecting the appropriate treatment option depends on the malocclusion, the anterior relationship, specific space requirements, and the condition of the adjacent teeth. The ideal treatment is the most conservative alternative that satisfies individual esthetic and functional requirements. This article closely examines the three options when replacing a missing lateral incisor with a tooth supported restoration. These options are a resin-bonded fixed partial denture, a cantilevered fixed partial denture, and a conventional full-coverage fixed partial denture. The specific criteria that must be evaluated for each option is addressed to illustrate the importance of interdisciplinary treatment planning to achieve optimal esthetics and long-term predictability. This article is the second of a three-part series discussing the three treatment alternatives for replacing congenitally missing lateral incisors. PMID- 16036124 TI - Direct restoration of worn maxillary anterior teeth with a combination of composite resin materials: a case report. AB - Tooth loss, alterations on tooth structure, and reduced vertical dimension are known to severely compromise the stomatognathic system. This case report describes the treatment of a patient who presented with an extremely worn maxillary anterior dentition with a loss of posterior support owing to the loss of almost all the posterior teeth, except the mandibular premolars. Provisional removable partial dentures were used to create an optimum maxillomandibular relationship and to provide restorative space prior to the restoration of the remaining teeth. This restoration was accomplished with a combination of layered hybrid and microfilled composite materials, which restored the maxillary anterior teeth to optimum esthetics and function. PMID- 16036125 TI - Fluorescence of layered resin composites. AB - BACKGROUND: Fluorescence is the absorption of light by a substance and the spontaneous emission of light in a longer wavelength within 10(-8) seconds of activation. PURPOSE: The purpose of this study was to determine the differences in fluorescence among layered resin composites with a color-measuring spectrophotometer. MATERIALS AND METHODS: Spectral reflectance and color of five brands of resin composites were measured over a white standard tile according to the Commission Internationale de l'Eclairage (CIE) CIELAB color scale relative to the standard illuminant D65. Human dentin was used as a control; five specimens were made for each group and each specimen was measured three times. An ultraviolet (UV) filter was inserted or removed to exclude or include the UV component of illumination. From the spectral reflectance values, subtraction spectrum by the inclusion and exclusion of the UV component was calculated. RESULTS: Dentin showed a fluorescence peak around 440 to 450 nm. Three of the five resin composites showed fluorescence peaks, and the peak wavelength was 440 to 450 nm. Peak height and peak area varied by the composite. Changes in color (DeltaE*(ab)) caused by the UV component were 1.58 to 2.35, and Deltab* values were from -2.20 to -1.49 in composites that showed fluorescence peaks. PMID- 16036126 TI - Effect of surface roughness on stain resistance of dental resin composites. AB - BACKGROUND: Surface quality of dental restorations is one of the important factors that determine the success of the restoration. Unfortunately, exterior discoloration is still a problem for dental resin composites. PURPOSE: The purpose of the study was to investigate the influential factors on stain resistance of dental resin composites. MATERIALS AND METHODS: Filtek Supreme (nanocomposite), Filtek A110 (microfilled composite), Filtek Z250 (microhybrid composite), and Filtek P60 (microhybrid composite) (all products from 3M ESPE, St. Paul, MN, USA) were tested. Thirty-six specimens per material were prepared and randomly assigned to 6 groups. The specimens in 5 groups were polished against 1,000-, 1,200-, 1,500-, 2,000-, and 2,500-grit sandpaper, respectively. The specimens in a sixth group were polished with 2,500-grit or 1,200-grit sandpaper and used as controls. Surface roughness (Ra) and gloss of the specimens were measured with a profilometer and a glossmeter, respectively. Specimens were immersed in a coffee solution (control group in distilled water) and kept in a 37 degrees C incubator. Color was measured by a spectrophotometer at baseline, 3 days, 7 days, and 14 days, and color change (DeltaE*(ab)) was calculated. RESULTS: Coffee, material, Ra, and the interaction of material x Ra had statistically significant influences on the stain resistance of the dental composites tested. For most materials in coffee solution, time squared and Ra had significant effects on the discoloration. CONCLUSIONS: Coffee had a significant influence on discoloration of the dental resin composite materials tested. The different composites behaved differently in coffee solution. Discoloration increased as Ra increased for the composites tested, except with Filtek A110. The discoloration process was accelerated with time. PMID- 16036127 TI - Effect of a light-emitting diode on composite polymerization shrinkage and hardness. AB - PURPOSE: This study evaluated the effect of light-emitting diodes (LEDs) on polymerization shrinkage and bottom-to-top hardness ratios of composites. MATERIALS AND METHODS: Six LEDs (Elipar FreeLight, 3M ESPE, St. Paul, MN, USA; Versalux, Centrix, Shelton, CT, USA; Ultra-Lume LED2, Ultradent, South Jordan, UT, USA; Zap LED only, CMS-Dental/Soft-Core Texas, North Richland Hills, TX, USA; Zap dual light; and L.E. Demetron I, Kerr Manufacturing Inc., Orange, CA, USA) and a quartz-tungsten-halogen (QTH) light (Optilux 501, Kerr Manufacturing Inc.) were tested. Ten specimens each of a microhybrid (Point 4, Kerr Manufacturing, Inc.) and a hybrid (Filtek Z250, 3M ESPE, St. Paul, MN, USA) composite, measuring 2 mm thick by 5 mm in diameter, were polymerized using each of the lights. Linear shrinkage was recorded using a contactless displacement instrument. Ten specimens were also polymerized with each light to determine the Knoop hardness number (KHN) at the top and bottom surfaces. RESULTS: For the microhybrid, Zap dual light had the least volumetric shrinkage 2.08% (+/- 0.33) and Elipar FreeLight had the highest 3.02% (+/- 0.73). There was no significant difference (p < or = .05) in shrinkage for the LEDs when compared with the Optilux 501. The hybrid showed the least amount of shrinkage with the L.E. Demetron I, 1.42% (+/- 0.12), and the greatest with the Zap dual light, 2.47% (+/- 0.31). The Ultra-Lume LED2 (p < or = .05) and Zap LED and dual light (p < or = .001) had significantly greater shrinkage than did the Optilux 501. Zap LED had the lowest depth of cure with a bottom KHN of 11.46 (+/- 2.71) and 33.62 (+/- 3.57) for the microhybrid and hybrid, respectively. The L.E. Demetron I had the highest bottom hardness value for the microhybrid, with a value of 40.65 (+/- 1.50). The Optilux 501 had the highest bottom hardness value for the hybrid, with a value of 62.03 (+/- 0.82). The Zap LED and dual light and Versalux lights had significantly lower bottom-to-top hardness ratios than did the QTH (p < or = .001) with the microhybrid. CONCLUSIONS: There was no statistically significant difference in shrinkage for the microhybrid with any of the lights tested. The hybrid, however, showed significantly less shrinkage with the halogen compared with the Ultra-Lume LED2 and Zap LED and dual light. All LEDs had equal or lower bottom hardness values than did the QTH, except for L.E. Demetron I with the microhybrid. PMID- 16036128 TI - Scanning electron microscope analysis of internal adaptation of materials used for pulp protection under composite resin restorations. AB - PURPOSE: The aim of this study was to evaluate the interfacial microgap with different materials used for pulp protection. The null hypothesis tested was that the combination of calcium hydroxide, resin-modified glass ionomer, and dentin adhesive used as pulp protection in composite restorations would not result in a greater axial gap than that obtained with hybridization only. MATERIALS AND METHODS: Standardized Class V preparations were performed in buccal and lingual surfaces of 60 caries-free, extracted human third molars. The prepared teeth were randomly assessed in six groups: (1) Single Bond (SB) (3M ESPE, St. Paul, MN, USA); (2) Life (LF) (Kerr Co., Romulus, MI, USA) + SB; (3) LF + Vitrebond (VT) (3M ESPE) + SB; (4) VT + SB; (5) SB + VT; (6) SB + VT + SB. They were restored with microhybrid composite resin Filtek Z250 (3M ESPE), according to the manufacturer's instructions. However, to groups 5 and 6, the dentin bonding adhesive was applied prior to the resin-modified glass ionomer. The specimens were then thermocycled, cross-sectioned through the center of the restoration, fixed, and processed for scanning electron microscopy. The specimens were mounted on stubs and sputter coated. The internal adaptation of the materials to the axial wall was analyzed under SEM with x1,000 magnification. RESULTS: The data obtained were analyzed with nonparametric tests (Kruskal-Wallis, p < or = .05). The null hypothesis was rejected. Calcium hydroxide and resin-modified glass ionomer applied alone or in conjunction with each other (p < .001) resulted in statistically wider microgaps than occurred when the dentin was only hybridized prior to the restoration. PMID- 16036129 TI - Critical appraisal: adhesive-composite incompatibility, part I. AB - Apart from some questions related to the repairability of resin composite restorations, dentists have always assumed that methacrylate-based resins are compatible with each other. For example, there is no clinically relevant problem in using a microfilled composite to laminate a Class IV restoration made with a hybrid composite, even if they are not of the same brand or manufacturer. In the context of adhesive systems, we have always believed that resin composites, regardless of their type or composition, bond well to all types of bonding agents. However, unexpected debonding of self-cured, core buildup composites that had been bonded with single-bottle adhesive systems was reported about 5 years ago. Subsequent studies demonstrated that there were, indeed, compatibility problems between simplified adhesive systems and self- or dual-cured resin composites. Apparently, when such combinations are used, reduced bond strengths and subsequent failures at the resin-adhesive interface can occur because of adverse reactions between the acidic resin monomers, an integral part of the simplified adhesive systems, and the chemicals involved in the polymerization mechanism of the self- or dual-cured composites, particularly the basic tertiary amines. At least one research group has expanded the information on this issue by further investigating the mechanisms involved in this phenomenon. This group demonstrated that not only adverse chemical reactions but also the permeability of such simplified systems contribute to the compromised bonding. This issue has profound clinical implications in view of the wide use of self- and dual-cured composites as core buildup materials and in the bonding of indirect restorations and endodontic posts. Some of the most representative studies of this group are described in this Critical Appraisal. Part II will appear in the next issue of the Journal. PMID- 16036130 TI - Differential expression of components of the microRNA machinery during mouse organogenesis. AB - MicroRNA (miRNA)-mediated gene silencing has recently emerged as a major mechanism of gene expression regulation during development in a variety of species. Little is known, however, about the presence of components of miRNA machinery in mammalian organogenesis. In this study, we report that members of the Argonaute (Ago) gene family are expressed in restricted of the day 11.5 and 14.5 embryo, including the brain, neural tube, limb, lungs, and hair follicles. In the developing lung, we found expression of Ago1 and Ago2 localized to branching regions, in distal epithelium and mesenchyme, respectively. These were sites undergoing the most dynamic changes in gene expression and rapid remodeling. We show that Ago1 transcripts are enriched in neural structures at these stages, consistent with the reported role of Drosophila Ago1 in the development of the central nervous system. Our results suggest a role for miRNAs in organogenesis. PMID- 16036131 TI - Synoviocytes, not chondrocytes, release free radicals after cycles of anoxia/re oxygenation. AB - By oxymetry and electron paramagnetic resonance (EPR), we investigated the effects of repeated anoxia/re-oxygenation (A/R) periods on the respiration and production of free radicals by synoviocytes (rabbit HIG-82 cell line and primary equine synoviocytes) and equine articular chondrocytes. Three periods of 20 min anoxia followed by re-oxygenation were applied to 10(7)cells; O(2) consumption was measured before anoxia and after each re-oxygenation. After the last A/R, cellular free radical formation was investigated by EPR spectroscopy with spin trapping technique (n=3 for each cell line). Both types of synoviocytes showed a high O(2) consumption, which was slowered after anoxia. By EPR with the spin trap POBN, we proved a free radical formation. Results were similar for equine and rabbit synoviocytes. For chondrocytes, we observed a low O(2) consumption, unchanged by anoxia, and no free radical production. These observations suggest an oxidant activity of synoviocytes, potentially important for the onset of osteoarthritis. PMID- 16036132 TI - A novel mechanism for the modulation of the Ras-effector interaction by small molecules. AB - When proteins require different conformations for their biological function, all these functional states have to coexist simultaneously in solution. However, the corresponding Gibbs free energy differences are usually rather high and thus the conformation with lowest energy predominates in solution whereas the populations of the states with higher energy (excited states) are very small. A stabilization of these excited states can be used as a novel principle to influence the activity of proteins by small molecules. For a proof of this principle, we selected the Ras protein that was shown by (31)P NMR spectroscopy to exist in solution in at least two different conformational states in its GTP form. One of these states shows a drastically reduced affinity to effectors. With Zn(2+) cyclen we found a small molecule which selectively stabilizes the weak-binding state. It may serve as lead compound for the development of a new type of Ras inhibitors. PMID- 16036133 TI - Expression of mutated cationic trypsinogen reduces cellular viability in AR4-2J cells. AB - Mutations in the human cationic trypsinogen are associated with hereditary pancreatitis. The cDNA coding for human cationic trypsinogen was subcloned into the expression vector pcDNA3. The mutations R122H, N29I, A16V, D22G, and K23R were introduced by site directed mutagenesis. We constructed an expression vector coding for active trypsin by subcloning the cDNA of trypsin lacking the coding region for the trypsin activating peptide behind an appropriate signal peptide. Expression of protein was verified by Western blot and measurement of enzymatic activity. AR4-2J cells were transiently transfected with the different expression vectors and cell viability and intracellular caspase-3 activity were quantified. In contrast to wild-type trypsinogen, expression of active trypsin and mutated trypsinogens reduced cell viability of AR4-2J cells. Expression of trypsin and R122H trypsinogen induced caspase-3 activity. Acinar cells might react to intracellular trypsin activity by triggering apoptosis. PMID- 16036135 TI - Office management and procedures. PMID- 16036136 TI - Office management: personnel issues overview. PMID- 16036137 TI - The role of the accountant in your practice. PMID- 16036138 TI - Negotiating insurance contracts--is there any hope? PMID- 16036139 TI - Medical records management--steps toward a paperless environment. PMID- 16036140 TI - Coding and reimbursement: the lifeline of the urologist's office. PMID- 16036141 TI - Strategic planning and budgeting. PMID- 16036142 TI - Marketing your urologic practice--ethically, effectively, and economically. PMID- 16036143 TI - Retirement plans for physicians and their employees. PMID- 16036144 TI - General and cystoscopic procedures. PMID- 16036145 TI - Office-based prostate procedures. PMID- 16036146 TI - Office urologic ultrasound. PMID- 16036147 TI - Office urodynamics. PMID- 16036148 TI - Tuberculosis of the spine and spinal cord. AB - Tuberculosis remains an important problem in underdeveloped countries, especially in individuals under the age of 20 years. In developed countries, there was an increase of the disease due to several factors, especially the appearance of A.I.D.S. The arterial haematogenous seeding is the most important way to the propagation of the disease to the rachis. The bacillus mycobacterium tuberculosis is characterized by a slow growth, propention for an oxygen rich environment and the absence of proteolytic enzymes. These characteristics are responsible for the imaging particularities of tuberculous osteomyelitis. The diagnosis can be confirmed by the bacteriologic and histological studies from samples removed by punction biopsy. PMID- 16036149 TI - Evaluating fMRI methods for assessing hemispheric language dominance in healthy subjects. AB - We evaluated two methods for quantifying the hemispheric language dominance in healthy subjects, by using a rhyme detection (deciding whether couple of words rhyme) and a word fluency (generating words starting with a given letter) task. One of methods called "flip method" (FM) was based on the direct statistical comparison between hemispheres' activity. The second one, the classical lateralization indices method (LIM), was based on calculating lateralization indices by taking into account the number of activated pixels within hemispheres. The main difference between methods is the statistical assessment of the inter hemispheric difference: while FM shows if the difference between hemispheres' activity is statistically significant, LIM shows only that if there is a difference between hemispheres. The robustness of LIM and FM was assessed by calculating correlation coefficients between LIs obtained with each of these methods and manual lateralization indices MLI obtained with Edinburgh inventory. Our results showed significant correlation between LIs provided by each method and the MIL, suggesting that both methods are robust for quantifying hemispheric dominance for language in healthy subjects. In the present study we also evaluated the effect of spatial normalization, smoothing and "clustering" (NSC) on the intra-hemispheric location of activated regions and inter-hemispheric asymmetry of the activation. Our results have shown that NSC did not affect the hemispheric specialization but increased the value of the inter-hemispheric difference. PMID- 16036150 TI - Protoneus-sequence: extended fluid-attenuated inversion recovery MR imaging without and with contrast enhancement. AB - Fluid-attenuated inversion recovery imaging (=flair imaging) is widely used as primary screening sequence in various investigation protocols, due to its high lesion contrast and sensitivity in detection of parenchymatous and leptomeningeal disease. An additional increase of sensitivity for detection of lesions may be achieved by contrast-enhanced flair imaging. Based on flair imaging a dual-echo inversion recovery imaging sequence (=proton echo usage [=protoneus] - sequence) was developed, which could significantly extend the possibilities of conventional flair imaging. PMID- 16036151 TI - Leptomeningeal high signal intensity (ivy sign) on fluid-attenuated inversion recovery (FLAIR) MR images in moyamoya disease. AB - PURPOSE: There are a few reports on leptomeningeal high signal intensity (LMHI: ivy sign) on fluid-attenuated inversion-recovery (FLAIR) images in moyamoya disease, but the feature of this finding has not been completely understood. The purpose of this study was to characterize LMHI on FLAIR images in moyamoya disease and to assess usefulness of this finding in the diagnosis of moyamoya disease in conventional MR imaging. MATERIAL AND METHODS: MR imaging of 28 patients with moyamoya disease was retrospectively reviewed. The grade of LMHI on FLAIR images was classified as "absent," "minimal," "moderate" and "marked." Fifty-four hemispheres of 28 patients (2 patients had unilateral disease) were assessed for the frequency of visualization and distribution of LMHI. The correlations between LMHI on FLAIR images, moyamoya vessels on T1- and T2 weighted images and MR angiography findings were also analyzed. RESULTS: Moderate and marked LMHI was seen in 31 out of 54 hemispheres (57%). LMHI was seen more prominently in the frontal and parietal lobes than in the temporal and occipital lobes. Although there was a tendency for LMHI on FLAIR images to be prominent in groups with moderate and marked moyamoya vessels on T1- and T2-weighted images, there was no significant correlation. More prominent LMHI was observed in the hemispheres in which cortical branches of the middle cerebral arteries were poorly visualized on MR angiography. CONCLUSION: Leptomeningeal high signal intensity (ivy sign) on FLAIR images is predominantly seen in the frontal and parietal lobes. Because this sign can be seen in patients with unremarkable moyamoya vessels, LMHI is a useful sign in conventional MR imaging for the diagnosis of moyamoya disease. PMID- 16036152 TI - Stent-angioplasty of intracranial vertebral and basilar artery stenoses in symptomatic patients. AB - BACKGROUND: To present our two-center treatment results with stent angioplasty of intracranial vertebrobasilar stenoses in symptomatic patients. MATERIAL AND METHODS: Between 2001 and 2003, 21 patients with 22 stenoses, refractory to medical therapy, who underwent elective stenting of intracranial vertebrobasilar stenoses were retrospectively analyzed. All patients had ischemic events clinically referable to the stenoses. Only high-grade stenoses of at least 80% were treated. Clinical evaluation was done based on the modified ranking scale (MRS). RESULTS: In all cases, the stent deployment turned out to be technically successful and control angiography demonstrated the elimination of the high-grade stenoses. A minor residual stenoses was still detectable in six cases. According to the MRS, clinical outcome improved in the case of four patients, seemed unchanged in 14 and worsened in three. The clinical morbidity rate amounted to 14%. Clinical follow-up was available for 13 patients after 9 months with no recurrent symptoms and angiographic follow-up was available after 10 months. Re stenoses occurred in two cases without clinical symptoms. Retreatment was not done. CONCLUSION: According to our data, stent angioplasty for symptomatic intracranial vertebrobasilar stenoses can be a treatment alternative in case of recurrent symptoms despite medical therapy; particularly, for stenoses of type Mori A or B. PMID- 16036153 TI - Central venous stenosis in haemodialysis patients without a previous history of catheter placement. AB - OBJECTIVE: To evaluate dialysis history, imaging findings and outcome of endovascular treatment in six patients with central venous stenosis without a history of previous catheter placement. MATERIAL AND METHODS: Between April 2000 and June 2004, six (10%) of 57 haemodialysis patients had stenosis of a central vein without a previous central catheter placement. Venography findings and outcome of endovascular treatment in these six patients were retrospectively evaluated. Patients were three women (50%) and three men aged 32-60 years (mean age: 45 years) and all had massive arm swelling as the main complaint. The vascular accesses were located at the elbow in five patients and at the wrist in one patient. RESULTS: Three patients had stenosis of the left subclavian vein and three patients had stenosis of the left brachiocephalic vein. The mean duration of the vascular accesses from the time of creation was 25.1 months. Flow volumes of the vascular access were very high in four patients who had flow volume measurement. The mean flow volume was 2347 ml/min. One of three patients with brachiocephalic vein stenosis had compression of the vein by the brachiocephalic artery. All the lesions were first treated with balloon angioplasty and two patients required stent placement on long term. Number of interventions ranged from 1 to 4 (mean: 2.1). Symptoms resolved in five patients and improved in one patient who had a stent placed in the left BCV. CONCLUSION: Central venous stenosis in haemodialysis patients without a history of central venous catheterization tends to occur or be manifested in patients with a proximal permanent vascular access with high flow rates. Balloon angioplasty with or without stent placement offers good secondary patency rates in mid-term. PMID- 16036154 TI - Outflow protection filters during percutaneous recanalization of lower extremities' arterial occlusions: a pilot study. AB - PURPOSE: Filter devices are already employed for the protection of carotid, coronary and renal distal vascular bed during endovascular procedures. This is a pilot study investigating their feasibility, safety and distal emboli protection capability during recanalization of lower extremities' acute and subacute occlusions. MATERIALS AND METHODS: Study population included 16 patients, 11 with a subacute arterial occlusion and 5 with an acute episode. The Trap filter (Microvena, USA) and its successor the Spider filter (EV3, USA) were utilized. Subacute occlusions were dealt with standard angioplasty and stenting procedures, while acute ones were managed primarily with Angiojet rheolytic thrombectomy. Outflow arterial tree was checked angiographically in-between consequent procedural steps. Embolic material collected after filter recovery was analyzed histopathologically. Patients' follow-up was scheduled at 1 month. RESULTS: Seventeen filter baskets were applied in the recanalization of 16 target lesions in total. Mean length of the occluded segments was 6.1 (range: 2-15 cm; S.D. = 3.7 cm). Mean in situ time of the filters was 38.75 min (range: 20-60 min; S.D. = 12.71 min). Technical success rate of deployment and utilization of the filtration devices was 100% (17/17). Procedural success rate of the recanalization was 100% (16/16) without any clinical or angiographic evidence of periprocedural distal embolization. Macroscopic particulate debris was extracted from all the filters (17/17) containing fresh thrombus, calcification minerals, cholesterol and fibrin. Mean diameter of the largest particle per specimen was 1702.80 (range: 373.20-4680.00 microm; S.D. = 1155.12 microm). No adverse clinical events occurred at 1-month follow-up with 100% limb salvage (16/16). CONCLUSION: The application of outflow protection filters is safe, feasible and efficacious in hindering distal embolization complications and safeguarding the distal capillary bed. Nevertheless, this is a pilot study in a limited group. Further studies have to be contacted in order to provide evidence for a more general use of these devices. PMID- 16036155 TI - Evaluation of pulmonary nodules: comparison of a prototype dual crystal (LSO/NAI) dual head coincidence camera and full ring positron emission tomography (PET). AB - PURPOSE: To determine the concordance of a prototype dual head coincidence camera (LSO-PS) and full ring PET (BGO-PET) using (18)F-fluorodeoxyglucose (FDG) in the evaluation of pulmonary nodules (PNs). MATERIALS AND METHODS: Patients referred for evaluation of < or =3 PNs (< or =3 cm diameter) were prospectively studied on the same day with both BGO-PET and LSO-PS. Imaging was performed at 60 and 120 min after injection of 370MBq FDG, respectively. Images were independently interpreted by four observers with each observer blinded to the other modality for the same patient. Lesions were scored in terms of relative intensity versus background. Non-attenuation corrected (nonAC) BGO-PET was used as the reference test. RESULTS: Forty-seven patients with 54 PNs (mean diameter 1.7 cm, S.D. 0.7) were included. Twelve nodules were in the < or =1.0 cm - 27 in the 1.1-2.0 cm - and 15 in the 2.1-3.0 cm range. Interobserver agreement was similar for both FDG imaging modalities. Using a sensitive assessment strategy with LSO-PS (> or = faint intensity deemed positive), there was a 97% (38/39, 95%CI 87-100%) concordance with BGO-PET and one false positive case with LSO-PS. Conservative reading (moderate or intense intensity deemed positive) resulted in a 92% (36/39, 95%CI 80-97%) concordance with BGO-PET, without false positives. The only lesion missed by LSO-PS using both assessment strategies involved a nodule 1.5 cm diameter that demonstrated moderate increased FDG uptake on BGO-PET. CONCLUSION: Depending on the test positivity criteria, LSO-PS demonstrates a high concordance (92-97%) with nonAC BGO-PET for the characterization of pulmonary nodules. PMID- 16036156 TI - The study of diagnostic accuracy of chest nodules by using different compression methods. AB - BACKGROUND: The purpose of this study was to compare the diagnostic accuracy of small nodules in the chest by using different compression methods. METHOD: Two radiologists with 5 years experience twice interpreted 39 chest images by using lossless and lossy compression methods. The time interval was 3 weeks. Each time the radiologists interpreted one kind of compressed images. The image browser used the Unisight software provided by Atlastiger Company in Shanghai. The interpreting results were analyzed by the ROCKIT software and the ROC curves were painted by Excel 2002. RESULTS: In studies of receiver operating characteristics for scoring the presence or absence of nodules, the images with lossy compression method showed no statistical difference as compared with the images with lossless compression method. CONCLUSION: The diagnostic accuracy of chest nodules by using the lossless and lossy compression methods had no significant difference, we could use the lossy compression method to transmit and archive the chest images with nodules. PMID- 16036157 TI - Conventional versus digital mammography in the analysis of screen-detected lesions with low positive predictive value. AB - PURPOSE: To compare the performance of screen-film and digital mammography in the assessment of screen-detected breast lesions. MATERIALS AND METHODS: A series of 100 consecutive mammographic screen-detected lesions (65 masses, 6 architectural distortions, 29 microcalcifications) deserving diagnostic assessment and judged to have a low positive predictive value underwent screen-film mammography (SFM) and digital mammography by a Fuji computed radiography system (FCR) (double exposure, same view, without removing compression) of the corresponding breast. Three sets of images (SFM, hard copy and soft copy FCR) were read, blind of assessment outcome, by three experienced radiologists. For the three different imaging modalities a contrast-detail analysis, dose evaluation and diagnostic accuracy by means of ROC analysis were performed. At the end of the diagnostic workup all suspicious cases (20) underwent surgical biopsy and were histologically confirmed as cancers and the cases which were negative or benign at assessment (80) were followed up for a period of 12-20 months. During the follow-up period two more cases proved to be cancers at subsequent examinations. RESULTS: Contrast-detail analysis gives better image quality for FCR compared to SFM at the same delivered dose, whilst in ROC analysis the SFM (AUC 0.7158), hard copy FCR (AUC 0.7404) and soft copy FCR (AUC 0.7501) (chi(2)=1.30, p=0.5220) are equivalent. CONCLUSION: FCR has a diagnostic performance equivalent to SFM in the assessment of screen-detected lesions with a low positive predictive value for cancer and it may be safely included in routine screening practice. PMID- 16036158 TI - A study on two different CAD systems for mammography as an aid to radiological diagnosis in the search of microcalcification clusters. AB - OBJECTIVE: The aim of the present study was to evaluate the efficacy of two different computer aided detection (CAD) systems for mammography in improving radiological diagnosis in the search of microcalcification clusters. The CAD systems used are: the SecondLooktrade mark (CADx Medical Systems, Canada) commercial system and the CALMA (computer assisted library in MAmmography) research CAD system. Three radiologists were asked to read mammographic images with and without the support of the CAD systems. MATERIAL AND METHODS: Three radiologists with respectively 3, 5 and 7 years of practice in mammogram reading in an Italian public hospital analysed a dataset composed of 120 digitized mammograms of healthy subjects with no lesion (proven by a radiological follow up of at least 3 years) and 70 images of patients with malignant cluster of microcalcification (proven by histopathological examination) both with no CAD support as well as with the help of the SecondLooktrade mark system. After 3 months they were asked to observe the same digitized mammograms with the assistance of the CALMA system. The radiologists worked independently and were unaware of the final diagnosis. The values of the area A(z) under the ROC curve, diagnostic sensitivity, specificity, positive and negative predictive values, and diagnostic accuracy were evaluated with and without the support of the CAD systems. The reading time and qualitative evaluations of each radiologist were also reported. RESULTS: With the support of the two CAD systems an improvement in A(z) area was obtained ranging from 0.01 to 0.04. Sensitivity increased from +8.6 to +15.7% and specificity decreased from 0.8 to 4.2%. CONCLUSION: In our study, not conditioned by the dataset, the CAD systems as second reader produced an increase in overall sensitivity of up to 15.7%, with a little decrease in specificity of up to 4.2%. Based on these results both CAD systems might be used in the current practise to improve the sensitivity values of conventional reading (radiologist alone). The results of this study show that no significant differences exist in term of A(z), sensitivity and specificity between CALMA and CADx. PMID- 16036159 TI - Stereotactic vacuum-assisted breast biopsies in 500 women with microcalcifications: radiological and pathological correlations. AB - AIM: We compared radiological assessment with pathological diagnoses in 500 consecutive vacuum-assisted breast biopsies performed for microcalcifications. METHODS: Lesions were biopsied using a 11-gauge mammotome device. Before biopsy, microcalcifications were classified according to the Breast Imaging Reporting and Data System (BI-RADS). Histopathological and radiological diagnosis were compared. RESULTS: Histopathology revealed 333 (67%) benign lesions. Benign lesions were classified as BI-RADS 3 in 19%, and as suspicious in 35%. 167 lesions (33%) were malignant. Malignant lesions were classified as suspicious or highly suggestive of malignancy in 63%. Frequency of malignancy in BI-RADS categories 4 and 5 was 35% and 100%, respectively. In BI-RADS 3 microcalcifications, the malignancy frequency was 19%. The mammographic features with the highest positive predictive value for malignancies were pleomorphic morphology (42%) and a linear or segmental distribution (51%). The microcalcification morphology was not reliably able to predict malignancy. CONCLUSION: In this study, BI-RADS 3 microcalcification lesions had a malignancy rate that is higher than previously reported. Vacuum-assisted biopsy is useful in any indeterminate and suspicious microcalcifications and provides maximum information before any operative intervention. PMID- 16036160 TI - Degenerative disc disease of the lumbar spine: a prospective comparison of fast T1-weighted fluid-attenuated inversion recovery and T1-weighted turbo spin echo MR imaging. AB - OBJECTIVE: To compare fast T1-weighted fluid-attenuated inversion recovery (FLAIR) and T1-weighted turbo spin-echo (TSE) imaging of the degenerative disc disease of the lumbar spine. MATERIALS AND METHODS: Thirty-five consecutive patients (19 females, 16 males; mean age 41 years, range 31-67 years) with suspected degenerative disc disease of the lumbar spine were prospectively evaluated. Sagittal images of the lumbar spine were obtained using T1-weighted TSE and fast T1-weighted FLAIR sequences. Two radiologists compared these sequences both qualitatively and quantitatively. RESULTS: On qualitative evaluation, CSF nulling, contrast at the disc-CSF interface, the disc-spinal cord (cauda equina) interface, and the spinal cord (cauda equina)-CSF interface of fast T1-weighted FLAIR images were significantly higher than those for T1 weighted TSE images (P<0.001). On quantitative evaluation of the first 15 patients, signal-to-noise ratios of cerebrospinal fluid of fast T1-weighted FLAIR imaging were significantly lower than those for T1-weighted TSE images (P<0.05). Contrast-to-noise ratios of spinal cord/CSF and normal bone marrow/disc for fast T1-weighted FLAIR images were significantly higher than those for T1-weighted TSE images (P<0.05). CONCLUSION: Results in our study have shown that fast T1 weighted FLAIR imaging may be a valuable imaging modality in the armamentarium of lumbar spinal T1-weighted MR imaging, because the former technique has definite superior advantages such as CSF nulling, conspicuousness of the normal anatomic structures and changes in the lumbar spinal discogenic disease and image contrast and also almost equally acquisition times. PMID- 16036161 TI - Impact of high field (3.0 T) magnetic resonance imaging on diagnosis of osteochondral defects in the ankle joint. AB - OBJECTIVE: To evaluate high field magnetic resonance (MR) imaging for imaging of osteochondral defects. MATERIALS AND METHODS: Nine osteochondral defects were simulated in three cadaveric talus specimens using a diamond drill. All specimens were examined on a 1.0 T MR unit and a 3.0 T MR unit. A T2-weighted turbo spin echo (TSE) sequence with a 2 mm slice thickness and a 256 x 256 matrix size was used on both scanners. The visibility of the osteochondral separation and the presence of susceptibility artifacts at the drilling bores were scored on all images. RESULTS: Compared to the 1.0 T MR unit, the protocol on the 3.0 T MR unit allowed a better delineation of the disruption of the articular cartilage and a better demarcation of the subchondral defect. Differences regarding the visualization of the subchondral defect were found to be statistically significant (P<0.05). Differences with regard to susceptibility artifacts at the drilling bores were not statistically significant (P>0.05). The average SNR was higher using 3.0 T MRI (SNR=12), compared to 1.0 T MRI (SNR=7). CONCLUSION: High field MRI enables the acquisition of images with sufficient resolution and higher SNR and has therefore the potential to improve the staging of osteochondral defects. PMID- 16036162 TI - Assessment of the specific absorption rate and calibration of decoupling parameters for proton decoupled carbon-13 MR spectroscopy at 3.0 T. AB - A strategy for proton decoupled carbon-13 MR spectroscopy ((1H)-13C MRS) with a strong static magnetic field (3.0 T) in vivo was investigated. The proton decoupling improves the signal-to-noise ratio, however, the effect of the decoupling power on the human body, especially in strong magnetic fields, should be considered. In order to establish a technique for monitoring the metabolism of glucose in the liver using (1H)-13C MRS at 3.0 T, two phantom experiments were performed. To assess whether the decoupling energy conformed to SAR limits defined by the IEC, temperature rises inside an agar gel phantom were monitored during a (1H)-13C MRS experiment. Then, the decoupling conditions of a glucose solution phantom were systematically optimized with combinations of decoupling bandwidth and power. The reliability of this procedure was discussed in conjunction with IEC guidelines. PMID- 16036163 TI - Towards making the science of complementary and alternative medicine more 'direct'. PMID- 16036164 TI - Outcome and costs of homoeopathic and conventional treatment strategies: a comparative cohort study in patients with chronic disorders. AB - OBJECTIVES: To evaluate the effectiveness of homoeopathy versus conventional treatment in routine care. DESIGN: Comparative cohort study. SETTING: Patients with selected chronic diagnoses were enrolled in medical practice. INTERVENTIONS: Conventional treatment or homeopathy. OUTCOME MEASURES: Severity of symptoms assessed by patients and physicians (visual rating scale, 0-10) at baseline, 6 and 12 months and costs. RESULTS: The analyses of 493 patients (315 adults, 178 children) indicated greater improvement in patients' assessments after homoeopathic versus conventional treatment (adults: homeopathy from 5.7 to 3.2; conventional, 5.9-4.4; p=0.002; children from 5.1 to 2.6 and from 4.5 to 3.2). Physician assessments were also more favourable for children who had received homoeopathic treatment (4.6-2.0 and 3.9-2.7; p<0.001). Overall costs showed no significant differences between both treatment groups (adults, 2155 versus 2013, p=0.856; children, 1471 versus 786, p=0.137). CONCLUSION: Patients seeking homoeopathic treatment had a better outcome overall compared with patients on conventional treatment, whereas total costs in both groups were similar. PMID- 16036165 TI - The effect of the homeopathic remedies Arnica montana and Bellis perennis on mild postpartum bleeding--a randomized, double-blind, placebo-controlled study- preliminary results. AB - OBJECTIVE: To evaluate the effect of Arnica Montana and Bellis perennis on postpartum blood loss. DESIGN: Double blind, placebo-controlled, randomized, clinical trial. SETTING: Department of Gynecology, Shaare Zedek Medical Center, Jerusalem. INTERVENTIONS: Forty parturients were randomized to one of three groups: Arnica montana C6 and Bellis perennis C6 (n=14), Arnica montana C30 and Bellis perennis C30 (n=14), or double placebo (n=12). After 48 h the Arnica/placebo was halted, and patients continued the Bellis/placebo until cessation of lochia. MAIN OUTCOME MEASURES: Hemoglobin levels (Hb) at 48 and 72 h postpartum. RESULTS: At 72 h postpartum, mean Hb levels remained similar after treatment with homeopathic remedies (12.7 versus 12.4) as compared to a significant decrease in Hb levels in the placebo group (12.7 versus 11.6; p<0.05), in spite of less favorable initial characteristics of the treatment group. The mean difference in Hb levels at 72 h postpartum was -0.29 (95% CI 1.09; 0.52) in the treatment group and -1.18 (95% CI -1.82; -0.54) in the placebo group (p<0.05). CONCLUSION: Treatment with homeopathic Arnica montana and Bellis perennis may reduce postpartum blood loss, as compared with placebo. PMID- 16036166 TI - Replication study concerning the effects of homeopathic dilutions of histamine on human basophil degranulation in vitro. AB - BACKGROUND: Various investigators have observed significant effects of highly diluted histamine on human basophil degranulation in vitro, compared to corresponding water controls. However, active and inactive dilution levels differed in most studies. OBJECTIVE: We aimed to reproduce former studies with flow-cytometry using rigorously controlled experimental conditions to minimise confounding factors. METHODS: In seven independent experiments, basophils of the same human donor were incubated with diluted histamine (up to 10(-34)M) or water controls and activated with anti-IgE antibodies. Basophil activation was determined by using bi-colour flow-cytometry. Experiments were blinded and performed with a randomised arrangement of the solutions on microtiter-plates. RESULTS: Histamine at the dilutions 10(-2)M and 10(-22)M was associated with a significant inhibition of basophil degranulation (p=0.018, Wilcoxon signed rank test) of 23.1% and 5.7%, respectively, if compared to "diluted" water treated in an identical manner. However, if all controls were pooled, only histamine 10(-2)M had a significant effect. Significant effects were seen for row numbers of the microtiter plates. CONCLUSION: We were not able to confirm the previously reported large effects of homeopathic histamine dilutions on basophil function of the examined donor. Seemingly, minor variables of the experimental set up can lead to significant differences of the results if not properly controlled. PMID- 16036167 TI - Challenges in herbal research: a randomized clinical trial to assess blinding with ginger. AB - OBJECTIVE: To assess methods to blind study participants to encapsulated ginger (Zingiber officinale). DESIGN: A randomized double-blind placebo controlled trial. SUBJECTS: Eighty healthy male and female volunteers. OUTCOME MEASURES: Whether participants can accurately determine if they receive a ginger or placebo capsule and a bottle filled with ginger or placebo capsules. RESULTS: Forty-two subjects correctly identified the capsule they received. Of those who received placebo, over 82% correctly identified their capsule. Only 22.5% of those who received ginger correctly identified their capsule. The likelihood of guessing ginger between the groups was statistically similar (p<0.01). 65% correctly guessed which bottle they had received (p=0.0073). Participants receiving the bottle filled with ginger capsules successfully identified their bottle 75% of the time (p=0.0016) compared to the 55% of the placebo group (p=0.5). CONCLUSIONS: Volunteers cannot determine which type of individual capsule they receive but can distinguish a bottle filled with ginger capsules. PMID- 16036168 TI - The psychological well-being and self-efficacy of carers of children with disabilities following attendance on a simple massage training and support programme: a 12-month comparison study of adherers and non-adherers. AB - OBJECTIVES: The Training and Support Programme (TSP) is an 8-week programme in which carers of children with disabilities receive instruction in simple massage techniques to use with their child. The aims of the present study were firstly to compare, adherers and non-adherers on measures of psychological well-being and self-efficacy and secondly, to examine whether, for adherers, the positive benefits of attending the TSP reported immediately after the Programme were maintained at 12-month follow-up. DESIGN: Eighty-two carers took part in the study. Data were collected 12-months after completion of the TSP by self-report questionnaires mailed to carers. For comparisons between adherers and non adherers at 12-month follow-up a between-groups design was used. For comparisons over time, a within-subjects design was used. RESULTS: Adherers had significantly higher levels of self-efficacy for managing their child's psychosocial well being, self-efficacy for carrying out the massage, and significantly better psychological well-being at follow-up compared to non-adherers. Furthermore, there were no significant differences over time on self-efficacy for managing their child's psychosocial well-being, self-efficacy for giving massage and levels of anxiety and depression at 12-month follow-up for adherers, suggesting that their improvements noted immediately post-programme were maintained at follow-up. Significant decreases on self-efficacy and depression were noted for non-adherers and there was a trend towards deterioration in anxious mood. CONCLUSION: In conclusion, the present study suggests that the positive benefits the TSP has for carers of children with disabilities can be maintained if carers continue to practise the massage at home with their child. PMID- 16036169 TI - Working towards a model of integrative health care: critical elements for an effective team. AB - OBJECTIVE: The objective of this study was to assess what factors the Tzu Chi Institute (TCI) practitioners identified as supports and barriers to providing care within an integrative health care setting. DESIGN: Qualitative data were collected by means of in-depth, semi-structured interviews with 16 practitioners who worked at a comprehensive integrative care clinic in Vancouver, Canada over 5 years. RESULTS: Practitioners identified four elements critical to forming and sustaining an effective integrative care team: (1) effective communication tools, (2) personal attributes (3) satisfactory compensation, and (4) a supportive organizational structure. CONCLUSIONS: Because of the influence practitioners have on the outcomes of patients in an integrative care program, attention must be focused not only on creating optimal healing environments for patients but also towards establishing and nurturing optimal working environments for practitioners. PMID- 16036170 TI - Lack of standards in informed consent in complementary and alternative medicine. AB - OBJECTIVES: To examine whether U.S.-based professional complementary and alternative medicine (CAM) organizations have explicit informed consent policies. DESIGN AND SETTING: Phone survey conducted in July-August 2003. RESULTS: Data were obtained from 37 out of a total of 39 eligible and accessible organizations representing 19 commonly used provider-administered CAM techniques (95% response rate.) Only 21 organizations (57%) had any informed consent policy and standards, whilst only six (16%) mandate their clinician members to routinely obtain an informed consent. CONCLUSIONS: We found no consistent standards with respect to the practice of informed consent across a broad range of CAM practices. CAM information that is not communicated appropriately or is otherwise unavailable to patients during the decision-making process may increase the potential for healthcare oversights and adversity. PMID- 16036171 TI - 'She did, he said': the use of ethnography in CAM research. PMID- 16036172 TI - Measuring health in quantitative studies. AB - For quantitative studies it is essential to select (and sometimes even construct) outcome measures, which are appropriate for measuring the specific aspect of health you are interested in. The measures should be valid, repeatable, and sensitive to change. It is complex and time-consuming to develop outcome measures afresh, so choose existing measures whenever possible and use them in the way they were intended. Measures may be generic or specific and should match the subjects involved. Objective measures, though accurate, may be less relevant to patients than subjective measures, particularly for CAM therapies. Findings made with one measure are more credible if they are supported by a second, as are results produced with the same measures used to measure outcomes in conventional medical studies. Careful planning of the analysis will help reveal any problems with measures. PMID- 16036173 TI - Developing a measure of treatment beliefs: the complementary and alternative medicine beliefs inventory. AB - OBJECTIVES: The study aimed to develop a comprehensive generic measure of treatment beliefs, the Complementary and Alternative Medicine Beliefs Inventory (CAMBI), and to identify distinct dimensions of CAM-related beliefs. METHODS: The CAMBI and other measures were presented and advertised online. RESULTS: 328 completed questionnaires were received. Factor analysis indicated three dimensions of beliefs could be identified. Subscales of the CAMBI were developed measuring beliefs in natural treatment, participation in treatment and holistic health. The subscales all had satisfactory reliability and were significantly correlated with CAM use (Spearman's rho=.18, .47 and .22 for natural treatments, holistic health and participation in treatment, respectively). CONCLUSIONS: The CAMBI measures three distinct dimensions of treatment beliefs, all of which are related to CAM use. PMID- 16036174 TI - Non-sleep effects of hypocretin/orexin. AB - Although a role for hypocretin/orexin (HCT/ORX) in sleep/wakefulness and arousal is widely recognized, other actions, not necessarily related to sleep, have been identified. Neurons producing the peptides project to brain sites known to be important in neuroendocrine and autonomic function, as well as appetite regulation. There is consensus that HCT/ORX plays a role in the regulation of cardiovascular function via its effects on sympathetic nervous activity, and the reported pharmacologic effects have been demonstrated to be physiologically relevant. Equally provocative are the actions of these peptides in the hypothalamus and pituitary gland to regulate reproductive and stress hormone secretion. While HCT/ORX are less potent stimulators of food intake than other hypothalamic peptides, HCT/ORX may play an integral role in the organization of hunger and satiation behaviors because of their interaction with those other peptides. In fact recent discoveries of interactions of HCT/ORX with peptides such as corticotropin releasing hormone and neuropeptide Y, as well as with aminergic neurotransmitter systems, are now defining the cellular and molecular mechanisms by which these potent neuropeptides act and promise insight into their physiologic relevance in a variety of non-sleep related behaviors and other homeostatic mechanisms. PMID- 16036175 TI - Rapid discrimination of rabies viruses isolated from various host species in Brazil by multiplex reverse transcription-polymerase chain reaction. AB - Rabies is carried mainly by mammalian carnivores and vampire bats in Latin America. However, rabies virus (RV) has been isolated in recent years from not only vampire bats in rural areas but also from several non-vampire bat species in urban areas, respectively. Therefore, rapid molecular screening is necessary for efficient epidemiology of these RVs. In this study, we investigated the usefulness of multiplex reverse transcription-polymerase chain reaction (RT-PCR) for determining the origins of 54 RV isolates from various host species in Brazil. And to evaluate the multiplex RT-PCR as a potential diagnostic tool, we investigated the sensitivity of this method. In addition, we compared the results with a phylogenetic tree developed from sequences of the RV glycoprotein (G protein) gene. Multiplex RT-PCR products showed five different sizes of products, whereas the phylogenic tree showed six groups. Of these six groups, four corresponded with the four sizes of the multiplex RT-PCR products. The other two groups showed correspondance with another one size of the multiplex RT-PCR products, indicating that multiplex RT-PCR results reflected the lineage of the 54 isolates. This study also showed that this method can detect trace amounts of RNA. In conclusion, this multiplex RT-PCR method allows the rapid, specific, and simultaneous detection of RVs isolated from various host species in Brazil. PMID- 16036176 TI - Analysis of two imported cases of yellow fever infection from Ivory Coast and The Gambia to Germany and Belgium. AB - BACKGROUND: Yellow fever remains one of the great burdens for public health in the endemic regions in Africa and South America. The under reporting of yellow fever cases in the respective regions and lack of international interest leads to an underestimation of the constant danger in these areas. Non-vaccinated travelers take a high risk without the effective protection of YFV 17D vaccination. OBJECTIVES: Two YF cases were imported to Europe in the last 4 years. We characterized two yellow fever virus (YFV) isolates from severely infected patients coming back from Africa, Ivory Coast and The Gambia, by genome sequencing and phylogenetic analysis. STUDY DESIGN: The virus infections in different organs were analyzed with pathological, immunohistological, electronmicroscopical and quantitative real-time PCR methods. RESULTS AND CONCLUSION: High virus loads in spleen and liver (2.4 x 10 (6) to 3 x 10 (7)GE/mL) demonstrated by real time PCR show massive virus replication leading to extraordinary progression of the disease in these patients. Immunohistological and electronmicroscopical analysis confirms virus particles in liver tissue. In all other organs no virus could be detected. A fast, specific and sensitive virus PCR detection is recommended for diagnostic of acute infections. The further sequence alignments show that the new isolates belong to the type II West African strain with great homology to over 40-year old YF isolates from Senegal and Ghana. The divergence observed was on average 3.3%, ranging from 0.0% to 5.0% in the coding region of Gambia 2001 strain and 2.9 %, ranging from 0.0% to 4.3% in the coding region of the Ivory C 1999 strain. Most mutations (5.0%/4.3%, respectively) occurred in the envelope protein. PMID- 16036177 TI - Atypical squamous cells of undetermined significance-favour reactive compared to atypical squamous cells of undetermined significance-favour dysplasia: association with cervical intraepithelial lesions and human papillomavirus infection. AB - BACKGROUND AND OBJECTIVES: The current study compared the cervical cytological sub-category "atypical squamous cells of undetermined significance-favour reactive (AFR)", recently recommended to be eliminated by the Bethesda system, to the sub-category "atypical squamous cells of undetermined significance-favour dysplasia (ASC-US)", in terms of prevalence of coexistent squamous intraepithelial lesions of either low-grade (LSIL) or high-grade (HSIL) and rate of human papillomavirus (HPV) infection. STUDY DESIGN: One hundred women with AFR and 100 with ASC-US were consecutively included in the study. All patients underwent colposcopy, followed by biopsy when necessary, and were screened for HPV infection by the combined use of Hybrid Capture II (DIGENE) and PCR with MY09/11 primers, the latter followed by direct sequencing of the amplifications products for HPV genotyping. RESULTS: LSIL were detected in 5.6% of AFR and 18.5% of ASC-US (p=0.00812), HSIL only in 4.3% of ASC-US. HPV infection was diagnosed in 11.2% of AFR and 38.0% of ASC-US (p=0.00003); high-risk HPV types (namely, HPV 16, -18, -31, -66, -67 and -70) were found in 6.7% of AFR and 22.8% of ASC-US (p=0.00239). Evidence of HPV infection in absence of SIL was proven in 7.1% of AFR and in 22.5% of ASC-US (p=0.00622). CONCLUSION: The association of AFR with SIL and high-risk HPV infection is low but not inexistent. Thus, to avoid the risk of leaving some high-risk AFR patients untreated or without follow-up, it could be proposed to keep AFR as a cytological category and to triage it by HPV testing, similarly to what has been already recommended for ASC-US. PMID- 16036178 TI - The development of a quality assurance programme for HPV testing within the UK NHS cervical screening LBC/HPV studies. AB - BACKGROUND: Cervical screening by cytology is effective but lacks sensitivity. The addition of human papillomavirus (HPV) testing can improve the effectiveness of screening for early identification of cervical disease. As HPV testing represents a new technology, a quality assurance (QA) programme is necessary to confirm the accuracy of results. OBJECTIVE: Our main objective was to design a QA programme for use in the English NHS liquid-based cytology (LBC) and HPV Cervical Screening Pilot Study. Our second objective was to use the knowledge gained to design a QA scheme for future general use within cervical screening and HPV testing programmes. STUDY DESIGN: Four elements were included in the programme: provision of clinical samples of known HPV status for internal quality control (IQC), distribution of panels of unknown samples for external quality assessment (EQA), resubmission of aliquots of samples to the reference laboratory for repeat testing and resubmission to reference laboratory to check for transport problems. Three sites took part in the QA programme using PreservCyt medium and ThinPrep for LBC preparation. The assay used at test sites was HPV hybrid capture (hc2) while the quality assurance laboratory used a combination of hc2, in-house HPV polymerase chain reaction (PCR) tests and HPV linear array (LA). RESULTS: Four negative, three low positive and 11 positive pools were used in 22 distributions of IQC samples. Seven distributions each of five 'unknown' EQA samples were sent out. Over 400 samples underwent repeat testing. Discrepant samples were further assessed to provide an explanation. Inter- and intra-laboratory consistency was high as measured by Kappa statistics and 96% agreement for EQA samples was obtained. CONCLUSIONS: The validity of the QA programme was established and reproducibility in different lab settings was reassuring. These results support the use of hc2 as a potential screening test in diagnostic laboratories. The need for robust quality assurance of HPV testing in cervical screening programmes was confirmed and lessons learnt from this pilot study will be incorporated in future schemes. PMID- 16036179 TI - A proteomics analysis of cellular proteins associated with HBV genotype-specific HBX: potential in identification of early diagnostic markers for HCC. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common cancers in the world and causes approximately one million deaths every year. HCC is highly prevalent in Asia and closely associated with chronic hepatitis B virus (HBV) infection, with HBX protein playing a key role in the hepatocarcinogenic process. In addition, HBV genotypes B and C are clinically associated with different outcome of infection. Reliable biomarkers for early diagnosis and prognosis of HCC are needed for effective treatment. OBJECTIVES: We propose to establish a proteomics approach to identify cellular proteins associated with HBX of a particular genotype. STUDY DESIGN: Genotype-specific HBXs are used to transfect non-HCC Chang cells. Transfected cell extracts are analyzed by proteomics approach to identify cellular proteins associated with specific HBX. RESULTS: A number of cellular proteins have been found to be specifically associated with HBX of genotype B or C. CONCLUSIONS: Our results suggest that proteomics approach could be used to identify cellular proteins associated with HBV infection of specific genotype. Similar approach could be deployed in the identification of molecular markers for early diagnosis of HCC. PMID- 16036180 TI - Detection and quantification of human metapneumovirus in pediatric specimens by real-time RT-PCR. AB - BACKGROUND: Human metapneumovirus (h MPV), a recently identified virus, causes respiratory illness in children. OBJECTIVES: A real-time reverse transcription polymerase chain reaction (RT-PCR) assay was developed and used to detect and quantify h MPV in respiratory specimens. STUDY DESIGN: The quantitative RT-PCR assay amplified an approximately 70 base pair fragment from the h MPV fusion protein gene. The assay was validated and used to test respiratory specimens obtained from children seen at a hospital in Seattle, Washington, from December 2002 through May 2003. RESULTS: The assay detected 1000 h MPV copies/mL of specimen, did not detect 19 other respiratory viruses, and was able to detect and accurately quantify isolates from the four known h MPV genetic lineages in a proficiency panel of 20 previously tested samples. h MPV was detected in 52 (7.2%) of 719 pediatric respiratory specimens. The mean log10 copies/mL of h MPV in the 52 positive specimens was 7.67 (range=4.59-10.60). Children aged 7-12 months had a significantly higher h MPV prevalence (12.4%) than did children younger than 7 months (4.7%) (P<0.005). Children in this age group also had significantly higher levels of h MPV in their respiratory specimens (mean log 8.43 copies/mL) than did the younger children (mean log 6.93 copies/mL) (P=0.0025). CONCLUSIONS: The rapid real-time RT-PCR assay described here is a sensitive test for clarifying the epidemiology of and diseases associated with h MPV. PMID- 16036181 TI - High levels of discordance between sequencing and serological subtyping in a predominantly non-B subtype HIV-1 infected cohort. AB - Samples from 457 randomly selected HIV-1 infected patients attending King's College Hospital were analysed using a subtype specific enzyme immunoassay. All serotyped non-Bs that provided unambiguous sequence and for which sufficient sample was available (n=100), which included three serotyped subtype B samples were further analysed by env sequencing and subtyping using neighbour joining phylogenetic analysis, the NCBI Retrovirus Genotyping tool and the Los Alamos BLAST search tool. Of the serotyped viruses, 45% (n=204) samples were subtype B. Specifically serotyped non-B strains (n=130) accounted for 28% of the total, of which the largest proportion were subtype C (n=66). Twenty-seven samples (6%) were classified as non-B, 9% (n=40) were multiply-reactive and 12% were non reactive (n=56). Of the 100 samples subtyped by sequencing the majority were subtype C (n=32), followed by subtype A (n=20). There was little concordance between the two methods. Although a 100% match was found among the serotyped and sequenced non-B viruses (n=13), only 16 of the sequenced subtype C specimens matched the 29 obtained by serotyping. Of the 20 multiply-reactive samples analysed by serotyping, only 1 sample consisted of a subtype mixture by sequencing. Of the 14 serologically non-reactive samples analysed, all were successfully sequenced, with subtype B strains (57%) the most common. Sequencing 15 samples in both env and pol regions revealed differences in subtype assignment for the same sample in some cases. Only 1/6 env subtype A and 4/5 env subtype C samples were concordant in pol sequence subtype. Differences were also found in subtyping by the different methods used. The overall agreement between the three methods was 89%. Four out of 11 samples agreed between the phylogenetic and Los Alamos methods, 1/11 between phylogenetic and BLAST and 2/11 between Los Alamos and BLAST. PMID- 16036182 TI - Response of HIV positive patients to the long-term salvage therapy by lopinavir/ritonavir. AB - BACKGROUND: The cohort of 19 patients on LPV/r salvage regimen was followed for the period of up to 37.5 months. Patient's virologic response was evaluated with regard to the various baseline characteristics. RESULTS: A 73.7% of patients (14 out of 19) achieved viral suppression during the first three months of treatment, either complete (47.4%) or partial (26.3%). This effect was only transient in five cases (virologic rebound emerged after 9 months of treatment on average) and in nine cases the treatment was successful in the long-term analysis (HIV RNA plasma level still undetectable at 31st month of the therapy on average with maximum of 36 months). We analyzed the link between the virologic response and possible predictive factors of treatment efficiency, such as lopinavir mutation score, various individual mutations, previous PI exposure, etc. We also describe changes in the PR sequence associated with poor response to the salvage therapy to LPV/r. CONCLUSIONS: The results of LPV/r salvage therapy were encouraging. About 47% of patients from our study achieved stable suppression of viral replication for 31 months on average. LPV/r proved to be potent inhibitor despite unfavourable prognosis. PMID- 16036183 TI - HIV-1 superinfection is not a common event. AB - Evidence for human immunodeficiency virus type 1 (HIV-1) superinfection was investigated among a group of four previously HIV-1 infected transfusion recipients (and the four implicated HIV-1 infected donors) identified by the Transfusion Safety Study, and two groups of 4 and 5 Brazilian injection drug users, who consistently injected themselves using shared paraphernalia. To probe these cases for possible superinfection we used heteroduplex mobility analysis (HMA) of HIV-1 tat, a technique which is a reliable for establishing epidemiologic linkages and searching for minor strains in mixed infection settings. In all these cases with established, untreated HIV-1 infections, we were unable to detect HIV-1 superinfection, even though the involved individuals were at high risk for second strain acquisition. We therefore conclude that although superinfection can occur in a few cases, it is a rare event, and the vast majority of recombinant HIV-1s characterized to date resulted from acute coinfections, rather than superinfection. PMID- 16036184 TI - An outbreak of primary dengue infection among migrant Chinese workers in Singapore characterized by prominent gastrointestinal symptoms and a high proportion of symptomatic cases. AB - BACKGROUND: An outbreak of dengue occurred among Chinese migrant workers at a construction site in Singapore that was characterized by a number of unusual features. METHODS: Clinical and laboratory data were prospectively collected from workers who were hospitalized with fever. We compared the frequency of disease manifestations in these patients with historical data from other cases of dengue admitted to the same centre. A serological survey was performed at the construction site one month after the onset of the outbreak. RESULTS: Thirty-nine patients were admitted from the affected construction site with confirmed acute dengue infection in March and April 2002. Fever, headache and rash were common and occurred at similar frequency in outbreak patients and historical controls. Gastrointestinal manifestations were significantly more common in the outbreak patients compared with controls (nausea 68%, diarrhea 41%, hyponatremia 47% and elevated urea 20%). Of the 274 studied in the serological survey, 27 had serological evidence of acute dengue, of whom 24 (89%) had experienced a febrile illness in the previous month. The virus was cultured and identified as Dengue-2. Few patients had a positive IgG for dengue indicating that this was likely a primary infection. CONCLUSION: This outbreak of primary dengue infection was characterized by unusually prominent gastrointestinal symptoms and a high proportion of symptomatic transmission. Manifestations of dengue may be altered when outbreaks involve immunologically naive adults who have migrated to dengue endemic areas. PMID- 16036185 TI - Bowel endometriosis and mucocele of the appendix. PMID- 16036186 TI - Spontaneous uterine rupture at 27 weeks of pregnancy after laparoscopic myomectomy. PMID- 16036187 TI - Trocar-associated injuries and fatalities: an analysis of 1399 reports to the FDA. AB - Laparoscopic trocars, medical devices used to gain access into the abdominal cavity, are the most common device named in malpractice injury claims associated with laparoscopic procedures. As part of its ongoing adverse event reporting program, the U.S. Food and Drug Administration (FDA) requires manufacturers and user facilities to file a report whenever a device was or may have been a factor in a death or serious injury. The FDA collects data from these reports in its Manufacturer and User Facility Device Experience (MAUDE) database. This study presents an analysis of fatality and injury data on laparoscopic trocars found in MAUDE reports received from January 1, 1997, through June 30, 2002, including 31 fatal injury cases and 1353 reports on nonfatal injuries. Cholecystectomy was the procedure most frequently associated with both fatal and nonfatal trocar injuries. Most fatalities involved vascular injuries. All fatality reports that identified the trocar design involved either a shielded trocar (which has a retractable shield that covers the trocar blade before and after insertion to help protect abdominal and pelvic organs from inadvertent puncture) or an optical trocar (which allows laparoscopists to view the cutting tip as it penetrates the tissues). Narrative comments cited surgical technique, device problems, and patient characteristics as contributing factors. Among nonfatal injuries, a change in surgical management such as additional surgical procedure--primarily laparotomy--prolonged surgery, or aborted surgery was reported most frequently for vascular and hollow viscus injuries. Many reports did not identify the device model, surgical procedure, or event timing, limiting Food and Drug Administration (FDA) and manufacturer investigations into whether the device contributed to the event. The most common manufacturer conclusions indicated the trocar was not returned, and no conclusions could be drawn about the trocar's contribution to the event. Fatalities occur with procedures in which shielded trocars and optical trocars are used. Further study is needed to evaluate the high proportion of reports associated with laparoscopic cholecystectomy. Laparoscopists should retain for evaluation any devices implicated in patient injuries and should ensure that detailed information on adverse events is provided in adverse event reports to the FDA. The FDA's Manufacturer and User Facility Device Experience (MAUDE) database can be a valuable source for information on adverse outcomes associated with medical devices and, given an understanding of its limitation, provides researchers with a viable adjunct to published literature and litigation surveys for obtaining this information. PMID- 16036188 TI - Submucous myomas: a new presurgical classification to evaluate the viability of hysteroscopic surgical treatment--preliminary report. AB - STUDY OBJECTIVE: To develop a new preoperative classification of submucous myomas for evaluating the viability and the degree of difficulty of hysteroscopic myomectomy. DESIGN: Retrospective study (Canadian Task Force classification II-3) SETTING: University teaching hospitals. PATIENTS: Fifty-five patients who underwent hysteroscopic resection of submucous myomas. INTERVENTION: The possibility of total resection of the myoma, the operating time, the fluid deficit, and the frequency of any complications were considered. The myomas were classified according to the Classification of the European Society for Gynaecological Endoscopy (ESGE) and by our group's new classification (NC), which considers not only the degree of penetration of the myoma into the myometrium, but also adds in such parameters as the distance of the base of the myoma from the uterine wall, the size of the nodule (cm), and the topography of the uterine cavity. The Fisher's exact test, the Student's t test, and the analysis of variance test were used in the statistical analysis. A p value less than .05 in the two-tailed test was considered significant. MEASUREMENTS AND MAIN RESULTS: In 57 myomas, hysteroscopic surgery was considered complete. There was no significant difference among the three ESGE levels (0, 1, and 2). Using the NC, the difference between the numbers of complete surgeries was significant (p <.001) for the two levels (groups I and II). The difference between the operating times was significant for the two classifications. With respect to the fluid deficit, only the NC showed significant differences between the levels (p = .02). CONCLUSIONS: We believe that the NC gives more clues as to the difficulties of a hysteroscopic myomectomy than the standard ESGE classification. It should be stressed that the number of hysteroscopic myomectomies used in this analysis was low, and it would be interesting to evaluate the performance of the classification in a larger number of patients. PMID- 16036189 TI - Total laparoscopic hysterectomy: preoperative risk factors for conversion to laparotomy. AB - STUDY OBJECTIVE: To identify the preoperative factors affecting the risk of conversion to laparotomy during total laparoscopic hysterectomy (TLH) indicated for benign conditions (surgery performed in cases of genital prolapse and/or urinary stress incontinence was excluded). DESIGN: Retrospective comparative study (Canadian Task Force classification II-2). SETTING: University tertiary referral center for gynecologic endoscopic surgery. PATIENTS: Four hundred sixteen consecutive patients who underwent TLH during the first 5 years of our experience performing TLH. INTERVENTION: Total laparoscopic hysterectomy. MEASUREMENTS AND MAIN RESULTS: The rate of conversion to laparotomy was 7% (29 patients). Factors that were found to be independently related to the risk of conversion to laparotomy are the following: body mass index (adjusted OR 1.09; 95% CI 1.01-1.18); uterine width on transvaginal ultrasonography (US) between 8 and 10 cm (adjusted OR 4.01; 95% CI 1.54-10.45); uterine width on US greater than 10 cm (adjusted OR 9.17; 95% CI 2.74-30.63); lateral myoma measuring greater than 5 cm on US (adjusted OR 3.57; 95% CI 0.97-13.17); history of adhesion-causing abdominopelvic surgery (adjusted OR 2.92; 95% CI 1.23-6.94). CONCLUSION: Transvaginal US evaluation is essential before performing TLH. Awareness of the risk factors for conversion to laparotomy is essential for proper patient information and better selection of patients. PMID- 16036190 TI - Office hysteroscopic sterilization compared with laparoscopic sterilization: a critical cost analysis. AB - STUDY OBJECTIVE: To evaluate the actual cost difference in performing Essure hysteroscopic sterilization in the office compared with ambulatory surgery using laparoscopic sterilization in the operating room. DESIGN: Cost-comparison analysis (Canadian Task Force classification III). SETTING: University hospital and affiliated outpatient office. INTERVENTIONS: Hysteroscopic placement of Essure device in an office setting and laparoscopic tubal ligation for permanent sterilization. MEASUREMENTS AND MAIN RESULTS: The various costs associated with the two procedures at our institution were compiled, and a direct cost comparison was made. We used actual institutional costs of the procedures, not billing or reimbursement. We found laparoscopic tubal ligations to cost 3449 dollars compared with hysteroscopic placement of the Essure device that costs 1374 dollars yielding a 2075 dollars difference between the procedures. CONCLUSION: In our institution and in our experience, office-hysteroscopic placement of the Essure device is a more cost-effective method than laparoscopic tubal ligation. PMID- 16036191 TI - Hysteroscopic myomectomy in postmenopausal women. AB - STUDY OBJECTIVE: To review the findings in postmenopausal patients undergoing hysteroscopic myomectomy. DESIGN: Retrospective case-controlled study (Canadian Task Force classification II-2). SETTING: Private practice. PATIENTS: Eighteen women with postmenopausal bleeding and two asymptomatic women with abnormal ultrasounds. INTERVENTIONS: Hysteroscopic myomectomy in 19 patients, with concomitant destruction of the endometrium in 4 patients, and resectoscopic biopsy in 1 patient. MEASUREMENTS AND MAIN RESULTS: Three patients underwent subsequent gynecologic surgery. One had hysteroscopy to evaluate and remove an asymptomatic residual myoma found on ultrasound. A sarcoma was found in two of the symptomatic patients. Two patients underwent hysterectomy-one for a sarcoma and the other for a carcinoma of the cervix. One patient has had further postmenopausal bleeding. CONCLUSION: Women who have a submucosal myoma that becomes symptomatic in the menopausal period may be at increased risk for a sarcoma. PMID- 16036192 TI - Effect of bupivacaine after operative laparoscopic gynecologic procedures. AB - STUDY OBJECTIVE: To evaluate the effect of intraperitoneal installation and periportal infiltration of bupivacaine on postoperative pain and return of bowel function. DESIGN: A prospective, double-blind, randomized trial (Canadian Task Force classification I). SETTING: GATA School of Medicine, Department of Obstetrics and Gynecology, Reproductive Endocrinology Unit. PATIENTS: Eighty patients undergoing operative gynecologic laparoscopy. INTERVENTION: Periportal infiltration of local anesthesia with 10 mL 0.5% bupivacaine before incision and another 20 mL 0.5% bupivacaine diluted with 20 mL of saline or equal amount of physiologic saline injected into the peritoneal cavity at the end of the procedure. MEASUREMENTS AND MAIN RESULTS: Each patient recorded the severity of her pain on a visual analog scale (VAS) at 1, 6, 18, and 24 hours and the time of first bowel movement and first flatus after surgery. Seventy-seven patients completed the study (38 in the bupivacaine group; 39 in the control group). The severity of postoperative pain, as recorded on the VAS, was significantly less at 1, 6, 18, and 24 hours after surgery in the group receiving bupivacaine compared with those in the control group. The first bowel movement in the bupivacaine group occurred earlier than in the control group (284.80 +/- 31.62 min vs 453.23 +/- 33.08 min, p <.001); similarly, the first flatus occurred earlier in the bupivacaine group than in the control group (466.2 +/- 29.59 min vs 658.80 +/- 40.92 min p <.001). CONCLUSION: Intraperitoneal installation and periportal infiltration of bupivacaine decrease postoperative pain and hasten the return of bowel function. Both decreased postoperative pain and shortened duration for the return of bowel function are crucial for comfort and discharge of the patient. PMID- 16036193 TI - Preemptive analgesia installation during gynecologic laparoscopy: a randomized trial. AB - STUDY OBJECTIVE: To evaluate the efficacy of intraoperative infusion of bupivacaine solution for the relief of pain after operative gynecologic laparoscopy. DESIGN: Prospective, double-blind, randomized, controlled trial (Canadian Task Force classification I4). SETTING: Tertiary teaching hospital. PATIENTS: Ninety-one women aged 16 to 69 years who underwent gynecologic laparoscopic surgery from November 2002 through November 2003. INTERVENTIONS: Group A (n = 30): intraperitoneal infusion of a mixture of 10 mL of 0.5% bupivacaine (50 mg) with epinephrine (1:500) in 40 mL of Ringer's lactate solution postoperatively. Group B (n = 30): the same mixture solution infusion preoperatively and postoperatively (total 100 mg bupivacaine). Group C (n = 31): placebo. MEASUREMENTS AND MAIN RESULTS: Shoulder tip pain (STP), abdominal parietal pain (APP), and abdominal visceral pain (AVP) were recorded on a visual analog scale at 2, 4, 8, 16, and 24 hours postoperatively. A total of 79 patients fulfilled the study criteria. The overall incidence of STP was 60.8%. Abdominal visceral pain in group B was significantly less than in group C at 2 and 4 hours postoperatively (p = .011 and p = .010, respectively). No statistically significant difference was found in length of hospital stay, postoperative meperidine consumption, or side effects. CONCLUSION: Intraperitoneal bupivacaine administration both immediately after placement of trocars and at the end of surgery was found to be effective in reducing the intensity of AVP but not in reducing STP, APP, or postoperative analgesia consumption after nonadvanced gynecologic laparoscopic procedures. The duration of the analgesic effect of bupivacaine instilled into the peritoneal cavity did not exceed 8 hours and probably was not dose related. PMID- 16036194 TI - Laparoscopic-assisted vaginal hysterectomy with uterine artery ligation through retrograde umbilical ligament tracking. AB - STUDY OBJECTIVE: To evaluate the clinical outcomes of uterine artery ligation through retrograde tracking of the umbilical ligament (RUL) in laparoscopic assisted vaginal hysterectomy (LAVH). DESIGN: Prospective study (Canadian Task Force classification II-3). SETTING: University-affiliated hospital. PATIENTS: Two hundred twenty-five women with myomas or adenomyosis. INTERVENTION: Laparoscopic-assisted vaginal hysterectomy with uterine artery identification and ligation through RUL. MEASUREMENTS AND MAIN RESULTS: The median age of the patients was 46 years, and the median weight of the extirpated uteri was 340 g, with 26.2% of uteri weighing more than 500 g. The median operation time was 135 minutes, and the median blood loss was 50 mL. The median intramuscular meperidine requirements were 1 ampoule (50 mg), and the median hospital stay was 3 days. It took approximately 10 minutes from identification of the umbilical ligament to ligation of the uterine artery. Uterine weight of 500 g or more required a significantly longer operation time compared with uteri weighing less than 500 g (164 min vs 127 min median, p <.001), and there was more blood loss (100 mL vs 50 mL median, p <.001). There were no differences in the median intramuscular meperidine requirements or hospital stay between the two groups. No blood transfusion was needed in either group, even in patients with a uterine weight of more than 1000 g. By regression analysis, uterine weight was significantly related to blood loss and operation time. A linear relationship was found among uterine weight, operation time, and blood loss: Uterine weight = 87.589 + 1.881 x operation time + 0.48 x blood loss (R = 0.531, F = 35.694, degrees of freedom 184, p <.001). No complications related to RUL were observed, although two bladder injuries related to severe pelvic adhesion with endometriosis and previous cesarean section occurred. CONCLUSION: Minimal blood loss and a low complication rate were noted in LAVH by uterine artery ligation through RUL. This technique should be a valid approach, especially in patients in whom minimal blood loss must be achieved. PMID- 16036195 TI - Directed laparoscopic cryomyolysis for symptomatic leiomyomata: one-year follow up. AB - STUDY OBJECTIVE: To evaluate the long-term effectiveness of laparoscopic cryomyolysis as a minimally invasive technique for the treatment of symptomatic uterine myomas in menstruating women. DESIGN: Open, one-arm pilot study (Canadian Task Force classification II). SETTING: University-affiliated public hospital. PATIENTS: Twenty patients with symptomatic uterine myomas were treated with directed cryomyolysis. All had reported abnormal bleeding and/or pelvic pain/pressure and/or urinary frequency. Myoma diameters varied from 4 to 10 cm. INTERVENTION: One-year follow-up after laparoscopic-directed cryomyolysis. MEASUREMENTS AND MAIN RESULTS: Laparoscopic cryomyolysis was performed using the Her Option Cryoablation Unit (American Medical Systems, Minneapolis, MN). Patients were evaluated 1, 3, 6, 9, and 12 months after surgery. Power color Doppler ultrasound was performed preoperatively and postoperatively to demonstrate the effectiveness of the technique in reducing or eliminating the primary blood supply to the myomas, as well documenting regression of the myomas. All patients reported a high rate of satisfaction with the treatment including absence of symptoms 12 months after surgery, with no bleeding and no myoma related symptoms, comparable with patients who underwent hysterectomy. Mean shrinkage of myoma volume increased until 9 months after surgery (59.5% +/- 13.2%), reaching a steady mean-volume reduction of approximately 60% (61.9% +/- 11.9%) 12 months after surgery. CONCLUSIONS: Directed laparoscopic cryomyolysis appears to be an effective and safe technique for providing rapid symptom relief and at least 12 months' effectiveness in the treatment of symptomatic uterine leiomyomas. PMID- 16036196 TI - Use of sublingual buprenorphine for pain relief in office hysteroscopy. AB - STUDY OBJECTIVE: To assess the efficacy of sublingual buprenorphine in the relief of pain associated with office hysteroscopy. DESIGN: Prospective, randomized study (Canadian Task Force classification I). SETTING: Tertiary medical center. PATIENTS: One hundred sixty-four women referred for office hysteroscopy from September 2003 through March 2004. INTERVENTION: Before hysteroscopy, 80 women received a tablet of buprenorphine (group A), and 84 women received a placebo (group B). Their pain sensations were evaluated on a 10-cm visual analog scale, and they were asked about the adverse reactions and level of satisfaction on the following day. MEASUREMENTS AND MAIN RESULTS: The pain score in group A was 3.3 +/- 1.1, which was similar to 3.2 +/- 1.3 in group B. The pain scores in subgroups of women also were similar within the same group and between the two groups. Thirty-one women (38.8%) in group A reported adverse reactions, including nausea, vomiting, and drowsiness, while none in group B reported any adverse reactions. CONCLUSION: Office hysteroscopy with a 3.1-mm flexible hysteroscope is a well-tolerated procedure. Sublingual buprenorphine is not helpful in relieving the pain associated with hysteroscopy but is associated with significant adverse reactions. PMID- 16036197 TI - Vascular atypia and irregularity on surface as signs of malignant adnexal mass: a complementary method of laparoscopic assessment. AB - STUDY OBJECTIVE: To evaluate the risk of malignancy associated with the main macroscopic findings observed on adnexal masses during a laparoscopic approach. DESIGN: Prospective series of consecutive patients (Canadian Task Force classification II-2). SETTING: Tertiary care university hospital. PATIENTS: One hundred fifteen patients were included. Ninety-eight had benign and 17 malignant adnexal masses. INTERVENTIONS: Diagnostic and operative laparoscopy. MEASUREMENTS AND MAIN RESULTS: Adnexal images were obtained during laparoscopy, recorded, and analyzed according to irregularity on surface, presence of atypical vessels, lobular form, absence of coral-like surface, implants, large-volume ascites, and cloudy or hemorrhagic ascites. A logistic regression was applied, and the independent findings associated with malignant disease were atypical vessels (OR 16.37; 95% CI 2.85-94.13; p <.01) and irregularity on tumor surface (OR 43.41; 95% CI 8.02-234.96; p <.05). In this way, 95% of the cases were correctly diagnosed. CONCLUSIONS: Atypical vessels and irregularity on tumor surface were important criteria of malignancy during laparoscopic treatment of adnexal masses. The recognition of these features can be helpful in choosing the ideal surgical approach mainly when the preoperative evaluation is unsatisfactory. PMID- 16036198 TI - Evaluation of a new surgical approach for the treatment of clomiphene citrate resistant infertility in polycystic ovary syndrome: laparoscopic ovarian multi needle intervention. AB - Laparoscopic ovarian drilling (LOD) is the accepted second-line treatment for clomiphene citrate-resistant anovulatory infertility in polycystic ovary syndrome (PCOS). Although multiple pregnancy rates are reduced with ovarian drilling procedures, postoperative adhesion formation is a potential complication in up to 85% of the women subjected to laparoscopic destructive ovarian procedures. Our objective was to determine the effectiveness of a new, specially designed laparoscopic device and technique that might enable treatment for patients with anovulatory PCOS with less trauma and fewer postoperative adhesions. Thirty-five infertile clomiphene citrate-resistant women with PCOS were included. Seventeen women underwent laparoscopic ovarian multi-needle intervention (LOMNI), and 18 women received step-up ovulation induction treatment with recombinant follicle stimulating hormone followed by intrauterine insemination for three cycles. Patients were followed for a period of 6 months after either laparoscopic surgery or the initiation of ovulation induction therapy. Outcome measures were cycle regularity, pregnancy rate, safety, postoperative adhesion formation, and cost effectiveness. There were no significant differences between the two groups in terms of age, body-mass index, duration of infertility, and basal cycle-day 2 hormone levels. Significant improvement in cycle regularity (p <.01) was found after LOMNI. Cumulative pregnancy rates (35.3% in the LOMNI group vs 33.3% in the ovulation induction group) did not differ between the groups. No adverse events following surgery were noted. Moderate ovarian hyperstimulation syndrome and multiple pregnancies occurred in four and two patients, respectively, in the ovulation induction group. Eight nonpregnant women in the LOMNI group underwent repeat laparoscopy at the end of the follow-up period. No adhesion formation attributable to LOMNI was observed in any of those eight women. The cost of LOMNI was significantly (p <.001) lower than the ovulation induction treatment. In conclusion, LOMNI may be a safe, inexpensive, and effective procedure for the treatment of CC-resistant infertility in patients with PCOS. It seems to preserve the beneficial effects and probably omits unwanted effects (such as adhesion formation) of LOD. PMID- 16036199 TI - Breast ductoscopy with a 0.55-mm mini-endoscope for direct visualization of intraductal lesions. AB - Standard radiologic examinations of breast duct lesions can give only indirect information. Mini-endoscopy with a breast ductoscope of only 0.55 mm offers direct visualization of the lesion and helps in the decision to perform or avoid exploratory breast tissue resection. We used a LaDuScope (PolyDiagnost, Pfaffenhofen, Germany) with a 0.55- or 0.95-mm outer diameter and a 75-mm working length from October 2003 through July 2004 on 11 women (average age of 48.3 years [range 36-69 years]) with suspicious nipple discharge. The optics have zero degree direct view, 70-degree field vision, and 3000 or 6000 pixel resolution. Breast ducts and walls could be easily inspected; and irrigation of breast ducts, aspiration, and use of cytology brush were possible under visual control. We had no intraoperative or postoperative complications. The new procedure of mini ductoscopy is feasible, safe, and helpful as an additional ambulatory diagnostic method for visual inspection of breast ducts. This instrument demonstrates the latest advances of technology and a trend toward less-invasive diagnostics for breast duct lesions. PMID- 16036200 TI - One hundred cases of laparoscopic subtotal hysterectomy using the PK and Lap Loop systems. AB - STUDY OBJECTIVE: To evaluate the safety and short-term outcomes of laparoscopic subtotal hysterectomy using the PK and Lap Loop systems. DESIGN: Prospective observational study (Canadian Task Force classification II-2). SETTING: Princess Royal University and Chelsfield Park Hospitals, Kent, UK. PATIENTS: One hundred women who underwent laparoscopic subtotal hysterectomy for menorrhagia from February 2003 through July 2004. INTERVENTION: The procedure was performed using the Plasma Kinetic (PK) system to seal the vascular pedicles and the Lap Loop system to separate the uterus at the level of the internal os. The uterus was removed from the abdominal cavity mainly by morcellation or posterior colpotomy. MEASUREMENTS AND MAIN RESULTS: Of 100 patients, 59 were operated on as outpatients. Mean patient age was 44.6 years, median parity was 2, mean body mass index was 26.8, and mean duration of symptoms was 4 years. Clinically, the uterus was enlarged in 70 patients, and preoperative ultrasound scanning suggested the presence of uterine myomas in 42 patients. In addition to hysterectomy, 47 patients had concomitant pelvic surgery. The mean total operating time was 45.5 minutes, and mean estimated blood loss was 114 mL. The overall major complication rate was 2%; two patients required blood transfusion after surgery. There were no bowel or urinary tract injuries, unintended laparotomy, return to operating room, or anesthetic complications. At follow-up, all patients were satisfied with surgery. CONCLUSION: Laparoscopic subtotal hysterectomy using the PK and Lap Loop systems for treatment of therapy-resistant menorrhagia is safe, can be performed as an outpatient procedure, and is associated with reduced operating time and high patient satisfaction. PMID- 16036201 TI - Suprapubic-assisted cystoscopic excision of intravesical tension-free vaginal tape. AB - We report on a 56-year-old woman with irritative voiding symptoms, which started 4 months after undergoing a TVT procedure for stress incontinence (SI). Evidence of the mesh coming through the left bladder wall did not emerge until cysto urethroscopic examination 2 years after surgery. Excision was done via transurethral cystoscopy with the aid of endoscopic scissors placed through a 5 mm suprapubic port inserted into the dome of the bladder, thus avoiding laparotomy. A Foley catheter was left in for 3 days, and after removal, the patient's symptoms disappeared almost completely. PMID- 16036202 TI - Laparoscopic removal of a remaining myoma after vaginal hysterectomy: a case report. AB - We report on an uncommon postoperative complication of vaginal hysterectomy--a remaining myoma--diagnosed and treated by laparoscopy. This procedure was performed for a remaining myoma 3 months after vaginal hysterectomy for fibroid uterus. Laparoscopy may be helpful to diagnose and treat this complication. PMID- 16036203 TI - Hysteroscopic management of cesarean scar pregnancy after unsuccessful methotrexate treatment. AB - Current management of cesarean scar pregnancy is conservative and consists of medical treatment with methotrexate (MTX). We present a report on a woman with this type of pregnancy who had continuous vaginal bleeding and persistent serum levels of beta human chorionic gonadotropin after undergoing curettage and MTX. Eventually, the remaining ectopic gestational tissue was removed by operative hysteroscopy. PMID- 16036204 TI - Laparoscopic management of a ruptured benign dermoid cyst during advanced pregnancy. AB - Benign cystic teratomas in pregnant women may be responsible for complications such as torsion, rupture and obstruction of labor. A woman in her 31st week of pregnancy with torsion of a large dermoid cyst and lipogranulomatosis peritonitis due to spilled cyst contents was managed laparoscopically with a favorable outcome. Trocar sites were selected according to the uterine size. Open laparoscopy allowed protection of the gravid uterus from penetrative injuries. Laparoscopic management of a voluminous adnexal mass may be safely performed during advanced pregnancy. PMID- 16036205 TI - Isobaric (gasless) laparoscopic myomectomy during pregnancy. AB - We report on the first case of an isobaric (gasless) laparoscopic myomectomy during the second trimester of pregnancy. Our patient had acute abdominal pain that did not respond to medical management. The procedure was performed under spinal anesthesia with conscious sedation. The remainder of the pregnancy was unremarkable. We believe that surgical management of uterine leiomyoma during pregnancy may be successfully performed in carefully selected patients. Laparotomy can be avoided, and pregnant patients can be managed safely by operative laparoscopy. With isobaric laparoscopy, the adverse effects and potential risks of CO2 insufflation are eliminated. The procedure can be performed under loco-regional anesthesia. The uterine closure can be performed safely and quickly as in laparotomy. PMID- 16036208 TI - Gap acceptance and risk-taking by young and mature drivers, both sober and alcohol-intoxicated, in a simulated driving task. AB - A single-blind randomized study was conducted on young (18-21 years, n = 16) and mature (25-35 years, n = 16) drivers to assess how age, combined with a modest dose of alcohol (0.7 g/kg for males and 0.6 g/kg for females), influenced performance on a driving simulator. The driving tasks included detecting the presence of a vehicle on the horizon as quickly as possible, estimating the point on the road that an approaching vehicle would have passed by the participants' vehicle (time-to-collision) and overtaking another vehicle against a steady stream of oncoming traffic. The results of the vehicle detection task showed that detection times were significantly slower with maturity, alcohol consumption and lower approaching vehicle speeds (50 kph), particularly on curved sections of road. Approaching vehicle speed was also found to significantly influence time-to collision (TTC) judgments, such that faster approach speeds led to less underestimated (and therefore riskier) judgments of TTC than slower speeds. In the overtaking task, mature participants demonstrated impaired discrimination skills with varying approaching vehicle speeds, while young participants recorded significantly slower speeds while overtaking a vehicle, thus increasing the time that they spent in the opposing lane. In conclusion, young and mature drivers demonstrated pivotal differences in behavior in this study. Young drivers showed a greater tendency to engage in risky driving, while experienced drivers appeared to be more susceptible to perceptual influences. Overall, alcohol consumption impaired a driver's ability to divide attention, but had little effect on decision-making processes. PMID- 16036209 TI - Evaluation of a program to reduce motor-vehicle collisions among young adults in the county of Landes, France. AB - The aim of this study was to evaluate the impact of a prevention program called "Atout-Route", based on the concept of commitment. The program was implemented in March 2000, to reduce the number of drivers under the age of 25 years involved in motor-vehicle collisions in the county of Landes (southwestern France). Using data from the regional observatory of road safety, we defined a target and three control groups similar on age or location. We used Poisson and quasi-Poisson regression to estimate whether the observed evolution of motor-vehicle collisions, after the program was implemented, was different in the targeted group than in the three control groups. The number of motor-vehicle collisions decreased everywhere and in every age group. The effect of the prevention program was not statistically significant (relative risk=0.89; 95% confidence interval 0.74-1.07). Our results are compatible with a positive effect of the program. The possible dilution of its effect by national road safety actions implemented since 2000, and our early assessment are possible reasons for the non-significant observation. PMID- 16036210 TI - Catching and matching bars with different orientations. AB - The hypothesis that perception enslaves action is examined by assessing whether systematic distortions in perceptual judgments are reflected by inaccuracies in catching. In the first experiment, participants had to align manually the orientation of a reference bar placed at different distances in the frontoparallel plane. In the second experiment participants had to catch differently orientated moving bars, which became invisible at different distances from the interception point. In the matching experiment, systematic errors in the alignment of orientation were found in particular for oblique orientations, the magnitude of which increased with increasing distance of the reference bar. The inaccuracies in the final hand orientation during the catching task, however, did not mirror this pattern of deviations. The findings are interpreted to be more consistent with recent views that vision for perception (i.e., matching) and vision for action (i.e., catching) are dissociated than with the view that perception enslaves action. PMID- 16036211 TI - Interactive imagery and colour in paired-associate learning. AB - In four experiments participants were instructed to imagine scenes that described either an animal interacting with a coloured object or scenes in which the animal and coloured object were independent of each other. Participants were then given the name of the animal and required to select the name of the object and its colour. The results showed that the classic interactive imagery effect was greater for the selection of the name of the object than it was for colour. In Experiments 2, 3, and 4, additional measures were taken which suggest that the effect for colour is dependent upon the retrieval of other features of the object (e.g., its form). Thus it is argued that there is no primary interactive imagery effect for colour. The results were predicted by a version of the shared information hypothesis. The implications of the results for alternative theories are also considered. PMID- 16036212 TI - Tissue factor mediates inflammation. AB - The role of tissue factor (TF) in inflammation is mediated by blood coagulation. TF initiates the extrinsic blood coagulation that proceeds as an extracellular signaling cascade by a series of active serine proteases: FVIIa, FXa, and thrombin (FIIa) for fibrin clot production in the presence of phospholipids and Ca2+. TF upregulation resulting from its enhanced exposure to clotting factor FVII/FVIIa often manifests not only hypercoagulable but also inflammatory state. Coagulant mediators (FVIIa, FXa, and FIIa) are proinflammatory, which are largely transmitted by protease-activated receptors (PAR) to elicit inflammation including the expression of tissue necrosis factor, interleukins, adhesion molecules (MCP-1, ICAM-1, VCAM-1, selectins, etc.), and growth factors (VEGF, PDGF, bFGF, etc.). In addition, fibrin, and its fragments are also able to promote inflammation. In the event of TF hypercoagulability accompanied by the elevations in clotting signals including fibrin overproduction, the inflammatory consequence could be enormous. Antagonism to coagulation-dependent inflammation includes (1) TF downregulation, (2) anti-coagulation, and (3) PAR blockade. TF downregulation and anti-coagulation prevent and limit the proceeding of coagulation cascade in the generation of proinflammatory coagulant signals, while PAR antagonists block the transmission of such signals. These approaches are of significance in interrupting the coagulation-inflammation cycle in contribution to not only anti-inflammation but also anti-thrombosis for cardioprotection. PMID- 16036213 TI - Inhibition of human plasma cholinesterase by malachite green and related triarylmethane dyes: mechanistic implications. AB - The inhibitory effects of the cationic triarylmethane (TAM+) dyes, pararosaniline (PR+), malachite green (MG+), and methyl green (MeG+) on human plasma cholinesterase (BChE) were studied at 25 degrees C in 100 mM Mops, pH 8.0, with butyrylthiocholine as substrate. PR+ and MG+ caused linear mixed inhibition of enzyme activity. The respective inhibitory parameters were K(i) = 1.9 +/- 0.23 microM, alpha = 13 +/- 48, beta = 0 and K(i) = 0.28 +/- 0.037 microM, alpha = 23 +/- 7.4, beta = 0. MeG+ acted as a competitive inhibitor with K(i) = 0.12 +/- 0.017 microM (alpha, infinity, beta, not applicable). The K(i) values were within the same range reported for a number of ChE inhibitors including propidium ion, donepezil, and the phenothiazines, suggesting that TAM+s are active site ligands. On the other hand, the alpha values failed to correlate with values previously reported for a number of ChE inhibitors. It appears that mixed inhibition is the combined result of more than one type of binding and S-I interference. The impact of ligands at the choline-specific and peripheral anionic sites (or, possibly, accessory structural domains) on BChE activity needs to be studied in further detail. PMID- 16036215 TI - EMP-1 is a junctional protein in a liver stem cell line and in the liver. AB - In an attempt to discover cell markers for liver stem cells, a cDNA microarray analysis was carried out to compare the gene expression profiles between an adult liver stem cell line, Lig-8, and mature hepatocytes. Several genes in the categories of extracellular matrix, cell membrane, cell adhesion, transcription factor, signal molecule, transporter, and metabolic enzyme were shown to be differentially expressed in Lig-8 cells. Among them, epithelial membrane protein (EMP)-1 has been previously implicated with stem cell phenotypes. Antiserum to EMP-1 was produced to localize its expression. On monolayers of Lig-8 cells, EMP 1 was expressed along the intercellular border. In the liver harboring proliferating oval cells, the liver progenitors, EMP-1 was localized as ribbon bands, a staining pattern for epithelial junctions, all the way through bile duct epithelia, oval cell ductules, and into peri-hepatocytic regions. These peri hepatocytic regions were proved to be bile canaliculi by co-localization of EMP-1 and dipeptidyl peptidase IV, an enzyme located on bile canaliculi. This report is the first to indicate EMP-1 to be a junctional protein in the liver. PMID- 16036214 TI - Calmodulin-dependent protein kinases phosphorylate gp130 at the serine-based dileucine internalization motif. AB - The receptor for leukemia inhibitory factor (LIF) consists of two polypeptides, the low affinity LIF receptor (LIFR) and gp130. We previously demonstrated that LIF stimulation caused phosphorylation of gp130 at Ser782, adjacent to a dileucine internalization motif, and that transient expression of a mutant receptor lacking Ser782 resulted in increased cell surface expression and increased LIF-stimulated gene expression compared to wild-type receptor. Phosphorylation of Ser782 on gp130 fusion protein by LIF-stimulated 3T3-L1 cell extracts was inhibited 61% by autocamtide-2-related inhibitory peptide (AIP), a highly specific and highly effective inhibitor of calmodulin-dependent protein kinase type II (CaMKII). Purified rat forebrain CaMKII was also able to phosphorylate gp130 fusion protein at Ser782 in vitro. Furthermore, antibodies targeting CaMKII and CaMKIV were able to immunoprecipitate gp130 phosphorylating activity from LIF-stimulated 3T3-L1 lysates. While pretreatment of cells with the MAPKK inhibitors PD98059 and U0126 blocked phosphorylation of Ser782 prior to LIF stimulation, these inhibitors did not block Ser782 phosphorylation by LIF stimulated 3T3-L1 cell extracts in vitro. These results show that CaMKII and possibly CaMKIV phosphorylate Ser782 in the serine-based dileucine internalization motif of gp130 via a MAPK-dependent pathway. PMID- 16036216 TI - The Troyer syndrome (SPG20) protein spartin interacts with Eps15. AB - The hereditary spastic paraplegias comprise a group of inherited neurological disorders in which the primary manifestation is spastic weakness of the lower extremities. Troyer syndrome is an autosomal recessive form of spastic paraplegia caused by a frameshift mutation in the spartin (SPG20) gene. Currently, neither the localization nor the functions of the spartin protein are known. In this study, we generated anti-spartin antibodies and found that spartin is both cytosolic and membrane-associated. Using a yeast two-hybrid approach, we screened an adult human brain library for binding partners of spartin. We identified Eps15, a protein known to be involved in endocytosis and the control of cell proliferation. This interaction was confirmed by fusion protein "pull-down" experiments as well as a cellular redistribution assay. Our results suggest that spartin might be involved in endocytosis, vesicle trafficking, or mitogenic activity, and that impairment in one of these processes may underlie the long axonopathy in patients with Troyer syndrome. PMID- 16036217 TI - Cdc2 and Cdk2 play critical roles in low dose doxorubicin-induced cell death through mitotic catastrophe but not in high dose doxorubicin-induced apoptosis. AB - In Huh-7 hepatoma cells, low dose (LD) doxorubicin treatment induces cell death through mitotic catastrophe accompanying the formation of large cells with multiple micronuclei, whereas high dose (HD) doxorubicin induces apoptosis. In this study, we investigated the role of Cdc2 and Cdk2 kinase in the regulation of the two modes of cell death induced by doxorubicin. During HD doxorubicin-induced apoptosis, the histone H1-associated activities of Cdc2 and Cdk2 both progressively declined in parallel with reductions in cyclin A and cyclin B protein levels. In contrast, during LD doxorubicin-induced cell death through mitotic catastrophe, the Cdc2 and Cdk2 kinases were transiently activated 1 day post-treatment, with similar changes seen in the protein levels of cyclin A, cyclin B, and Cdc2. Treatment with roscovitine, a specific inhibitor of Cdc2 and Cdk2, significantly blocked LD doxorubicin-induced mitotic catastrophe and cell death, but did not affect HD doxorubicin-induced apoptosis in Huh-7, SNU-398, and SNU-449 hepatoma cell lines. Our results demonstrate that differential regulation of Cdc2 and Cdk2 activity by different doses of doxorubicin may contribute to the induction of two distinct modes of cell death in hepatoma cells, either apoptosis or cell death through mitotic catastrophe. PMID- 16036218 TI - Stimulation of the creatine transporter SLC6A8 by the protein kinases SGK1 and SGK3. AB - Creatine binds phosphate thus serving energy storage. Cellular creatine uptake is accomplished by the Na+,Cl-, creatine transporter CreaT (SLC6A8). The present study explored the regulation of SLC6A8 by the serum and glucocorticoid inducible kinase SGK1, a kinase upregulated during ischemia. In Xenopus oocytes expressing SLC6A8 but not in water injected oocytes creatine induced a current which was significantly enhanced by coexpression of wild type SGK1 and constitutively active (S422D)SGK1, but not inactive (K127N)SGK1. Kinetic analysis revealed that (S422D)SGK1 enhanced maximal current without significantly altering affinity. The effect of SGK1 was mimicked by the constitutively active isoform (S419D)SGK3 but not by inactive (K119N)SGK3, wild type isoform SGK2 or constitutively active related kinase (T308D,S473D)PKB. In conclusion, the kinases SGK1 and SGK3 increase SLC6A8 activity by increasing the maximal transport rate of the carrier. Deranged SGK1 and/or SGK3 dependent regulation of SLC6A8 may affect energy storage particularly in skeletal muscle, heart, and neurons. PMID- 16036219 TI - Mechanical stress-dependent transcriptional regulation of sarcolipin gene in the rodent atrium. AB - Sarcolipin, a homologue of phospholamban, regulates Ca2+ uptake through the interaction with sarcoplasmic reticulum Ca2+ ATPase (SERCA) and is predominantly expressed in the atrial muscle. Although the atrial chamber-specific expression of sarcolipin could be primarily regulated at the transcriptional level, the transcriptional regulation remains poorly understood. Since mechanical stress plays an important role in transcriptional regulation of a gene involved in cardiac hypertrophy and remodeling, we generated left-sided or right-sided pressure-overload models by transverse aortic constriction (TAC) in ddY mice or by monocrotaline administration in Wistar rats, respectively. TAC significantly decreased the expression of sarcolipin, SERCA2a, and phospholamban mRNAs in the left atrium (LA) than those in the right atrium (RA). By contrast, monocrotaline administration significantly decreased the expression of sarcolipin, SERCA2a, and phospholamban mRNAs in the RA than those in the LA. The two independent complementary experiments unequivocally demonstrated that mechanical stress down regulates the transcription of the sarcolipin gene. PMID- 16036220 TI - CAND1 enhances deneddylation of CUL1 by COP9 signalosome. AB - Cullin-RING ligases (CRLs) regulate diverse cellular functions such as cell cycle progression and cytokine signaling by ubiquitinating key regulatory proteins. The activity of CRLs is controlled by Nedd8 modification of the cullin subunits. Recent reports have suggested that CAND1, which specifically binds to unmodified CUL1 but not to neddylated one, is required for the in vivo function of SCFs, the CUL1-containing CRLs. We show here that CAND1 and COP9 signalosome (CSN), the major deneddylase of cullins, bind to unneddylated CUL1 in a mutually exclusive way. The suppression of CAND1 expression by small inhibitory RNA enhanced the interaction between CUL1 and CSN, suggesting that CAND1 inhibited the binding of CSN to CUL1. We found that the binding of CSN to CUL1 required the four helix bundle in CUL1 C-terminal domain, which was wrapped around by CAND1 in the CAND1 CUL1-Rbx1 complex. CAND1 greatly facilitated CSN-mediated deneddylation of CUL1 in vitro, which was dependent on its binding to CUL1. Our data suggest that enhancement of CSN-mediated deneddylation by CAND1 may contribute to its function as a positive regulator of SCFs in vivo. PMID- 16036221 TI - Up-regulated expression of low molecular weight protein tyrosine phosphatases in different human cancers. AB - Protein tyrosine phosphorylation, mediated by the balanced action of tyrosine kinases and phosphatases, contributes to the regulation of the growth, migration, and invasion of normal and malignant cells. Among tyrosine phosphatases, low molecular weight protein tyrosine phosphatases (LMW-PTP) have been recognized as a possible "positive factor" in tumour onset and progression. The aim of this work was to assess whether LMW-PTP are differentially expressed in normal and malignant tissues. Using real-time PCR analysis we evaluated the expression levels of total LMW-PTP mRNA in surgical samples of breast, colon and lung cancers (63, 60, and 58, respectively), and in their paired adjacent not affected tissues. Moreover, the same analysis was carried out on a group of neuroblastomas (25 cases). Significant correlations between LMW-PTP overexpression and the most common clinical-pathological features of cancers exist. In colon cancer and neuroblastoma increased total LMW-PTP mRNA expression correlates with unfavourable outcome. While LMW-PTP mRNA expression increases in tumour samples, the relative contribution of the different isoforms does not change. Our findings indicate that LMW-PTP can be considered an oncogene as it is overexpressed in different tumour types and suggests that LMW-PTP enhanced expression is generally prognostic for a more aggressive cancer. PMID- 16036222 TI - Isolated mouse liver mitochondria are devoid of glucokinase. AB - Glucokinase is a hexokinase isoform with low affinity for glucose that has previously been identified as a cytosolic enzyme. A recent report claims that glucokinase physically associates with liver mitochondria to form a multi-protein complex that may be physiologically important in apoptotic signaling [N.N. Danial, C.F. Gramm, L. Scorrano, C.Y. Zhang, S. Krauss, A.M. Ranger, S.R. Datta, M.E. Greenberg, L.J. Licklider, B.B. Lowell, S.P. Gygi, S.J. Korsmeyer, Nature 424 (2003) 952-956]. Here, we re-examined the association of glucokinase with isolated mouse liver mitochondria. When glucokinase activity was measured by coupled enzyme assay, robust activity was present in whole liver homogenates and their 9500 g supernatants (cytosol), but activity in the purified mitochondrial fraction was below detection (<0.2% of homogenate). Furthermore, addition of 45 mM glucose in the presence of ATP did not increase mitochondrial respiration, indicating the absence of ADP formation by glucokinase or any other hexokinase isoform. Immunoblots of liver homogenates and cytosol revealed strong glucokinase bands, but no immunoreactivity was detected in mitochondria. In conclusion, mouse liver mitochondria lack measurable glucokinase. Thus, functional linkage of glucokinase to mitochondrial metabolism and apoptotic signaling is unlikely to be mediated by the physical association of glucokinase with mitochondria. PMID- 16036223 TI - Cloning and characterization of Munc18c(L), a novel murine Munc18c gene paralog. AB - We have identified and characterized a new mouse gene sequence, Munc18c(L), that appears closely related to the syntaxin-binding protein, Munc18c. The novel Munc18c(L) gene is comprised of 2 exons separated by a 600bp intron sequence with non-consensus donor and acceptor sites. Exons 1 and 2 of Munc18c(L) overlap with exons 1 through half of 9 of the Munc18c gene. The deduced amino acid sequence of Munc18c(L) is 271 amino acids long with homology to Munc18c protein ending at position 250. RT-PCR of murine tissues showed expression of Munc18c(L) in various tissues. RT-PCR carried out with a primer spanning the ATG codon and another one specific for the exon 2 of Munc18c(L) revealed two different transcripts of 0.8 and 1.4kbp in length. Using 5'-RACE, the start of Munc18c(L) exon 1 matches the one predicted for Munc18c, but the proximal promoter differ. This first identification of Munc18c(L) is vital in differentiating between Munc18c(L) and Munc18c and their potential roles in insulin-mediated glucose uptake. PMID- 16036224 TI - FimH adhesin of Escherichia coli K1 type 1 fimbriae activates BV-2 microglia. AB - The generation of intense inflammation in the subarachnoid space in response to meningitis-causing bacteria contributes to brain dysfunction and neuronal injury in bacterial meningitis. Microglia, the major immune effector cells in the central nervous system (CNS), become activated by bacterial components to produce proinflammatory immune mediators. In this study, we showed that FimH adhesin, a tip component of type 1 fimbriae of meningitis-causing Escherichia coli K1, activated the murine microglial cell line, BV-2, which resulted in the production of nitric oxide and the release of tumor necrosis factor-alpha. Mitogen-activated protein kinases, ERK and p-38, and nuclear factor-kappaB were involved in FimH adhesin-mediated microglial activation. These findings suggest that FimH adhesin contributes to the CNS inflammatory response by virtue of activating microglia in E. coli meningitis. PMID- 16036225 TI - The effects of desmethylimipramine on cyclic AMP-stimulated gene transcription in a model cell system. AB - The present study utilised an in vitro cell model of the cAMP signalling pathway to investigate the actions of desipramine (DMI) and other psychoactive agents on cAMP-driven gene transcription. The model comprised CHObeta2 SPAP cells; Chinese hamster ovary cells expressing human beta2 adrenoceptors and a secreted placental alkaline phosphatase (SPAP) reporter gene with multiple cAMP response elements (CREs) in its promoter region. SPAP assays showed DMI to inhibit isoprenaline or forskolin-enhanced gene transcription in a time and concentration-dependent manner (IC50=16.6+/-2.0 microM after 18 h). This effect of DMI was not dependent upon activity at the levels of the beta2 receptor, cAMP accumulation or phosphorylation of the transcription factor, cAMP response element binding protein (CREB). The inhibitory effects were maintained in the presence of DMI for at least 3 weeks and were mimicked by exposure to norfluoxetine (the major metabolite of fluoxetine; IC50=7.2+/-1.8 microM) and the neuroleptics, chlorpromazine and clozapine, all at a concentration of 10 microM. Amphetamine (10 microM, 18 h) enhanced SPAP gene transcription. Ca2+ imaging experiments ruled out an inhibitory effect of DMI on Ca2+ influx as concluded by previous studies. The results suggest a molecular target for DMI that lies downstream of CREB phosphorylation. Whether the inhibitory action of DMI is common to naturally expressed CRE-driven genes involved in adaptive responses to antidepressants in vivo remains to be determined. PMID- 16036226 TI - AIDS-related discrimination in Asia. AB - The Asia Pacific Network of People Living with HIV/AIDS (APN+) conducted the first regional documentation of AIDS-related discrimination in Asia. This project was an action-based, peer-implemented study that aimed to develop an understanding of the nature, pattern and extent of AIDS-related discrimination in several Asian countries. Trained HIV-positive people interviewed 764 positive people in four countries (India 302; Indonesia 42; Thailand 338; the Philippines 82) using a structured questionnaire. Findings indicate that the major area of discrimination in each country is within the health sector, where over half of those surveyed experienced some form of discrimination. In all countries, the majority of people did not receive pre-test counselling before being tested for HIV. People who reported coerced testing were significantly more likely than other respondents to face subsequent AIDS-related discrimination. A considerable number of respondents were refused treatment after being diagnosed with HIV and many experienced delayed provision of treatment or health services. Breaches of confidentiality by health workers were common. Within the family and the community, women were significantly more likely to experience discrimination than men, including ridicule and harassment, physical assault and being forced to change their place of residence because of their HIV status. These findings have serious implications, particularly in light of the increasing trend in many countries to test all pregnant women in order to prevent transmission of HIV to their unborn children. PMID- 16036227 TI - A method for the quantitative analysis of the layering of HIV-related stigma. AB - HIV-related stigma is regarded as one of the major barriers in the development of effective prevention and care programs; but the stigma associated with HIV stigma is not a singular entity. The stigma of the infection is layered with other stigmas, such as those associated with the routes of transmission (e.g., sex work and injecting drug use) and personal characteristics (e.g., race, religion, ethnicity and gender). In developing programs and policies to overcome HIV related stigma, cognisance needs to be taken of all the sources of stigma, and how they may interact. A novel method is described for examining the layers of HIV/AIDS-related stigma, and secondary data are adapted to illustrate this. The importance of understanding the layering of stigma for the development of effective interventions is also discussed. PMID- 16036228 TI - Predictors of sexual transmission risk behaviors among HIV-positive young men. AB - Reduction in the incidence of high-risk sexual behaviors among HIV-positive men is a priority. We examined the roles of proximal substance use and delinquency related variables, and more distal demographic and psychosocial variables as predictors of serious high-risk sexual behaviors among 248 HIV-positive young males, aged 15-24 years. In a mediated latent variable model, demographics (ethnicity, sexual orientation and poverty) and background psychosocial factors (coping style, peer norms, emotional distress, self-esteem and social support) predicted recent problem behaviors (delinquency, common drug use and hard drug use), which in turn predicted recent high-risk sexual behaviors. Hard drug use and delinquency were found to predict sexual risk behaviors directly, as did lower self-esteem, white ethnicity and being gay/bisexual. Negative peer norms strongly influenced delinquency and substance use and positive coping predicted less delinquency. In turn, less positive coping and negative peer norms exerted indirect effects on sexual transmission risk behavior through delinquency and hard drug use. Results suggest targeting hard drug use, delinquency, maladaptive peer norms, dysfunctional styles of escaping stress and self-esteem in the design of intervention programs for HIV-positive individuals. PMID- 16036229 TI - Home-based care for orphaned children infected with HIV/AIDS in Uganda. AB - The primary aim of this paper is to describe an outreach programme from a main state hospital in sub-Saharan Africa, which has been running for three years. This programme is based in Mulago Hospital, Kampala, Uganda and cares for up to 200 children infected with HIV/AIDS in their home. We describe the clinic and how we meet the families and enrol them, the infrastructure of the programme and the personnel involved. Children and their families receive physical, psychological and social care and we describe each aspect of this. The knowledge base about older children with AIDS in Africa is scarce and the secondary aim of this paper is to publish observations that were made while providing care. This includes demographics and the health problems encountered among children living with HIV/AIDS in a resource-poor setting who do not receive antiretroviral medication. Finally, we discuss the strengths and weaknesses of this model of care and the prerequisites to setting up a similar model. PMID- 16036230 TI - HIV testing and counseling among adolescents attending family planning clinics. AB - This study examined whether an assessment tool combining HIV-related risk behaviors and symptoms would increase HIV testing and return for post-test counseling among 466 adolescents attending family planning clinics. The results indicted that high-risk behaviors were common among these adolescents. The majority (78.3%) identified themselves as not using condoms consistently, almost a quarter (24.7%) reported a history of STDs and 129 (27.7%) reported they had pierced their bodies. A total of 214 (45.9%) received HIV testing. Two (0.4%) adolescents, one male and one female, tested positive for HIV. Of the 214 adolescents who underwent testing, 183 (85.5%) returned to the clinic for post test counseling. The results of this study indicated that adolescents who reported risk behaviors and symptoms were not more likely than those who reported no risk behaviors and symptoms to request testing and return for post-test counseling. In light of these results, the authors review the protocols associated with testing and post-test counseling and propose solutions that can potentially improve these processes. The authors recommend integrating a risk assessment tool with HIV testing in family planning clinics as testing and return for post-test counseling rates were high. PMID- 16036231 TI - Evaluating the sexual behaviour, barriers to condom use and its actual use by university students in Nigeria. AB - This research quantified the barriers to condom use of a sample of university students in Nigeria. It further analyzed association between condom use at last intercourse and the barriers, in addition to HIV risk sexual practices. A total of 96 women and 128 men were randomly selected, and they completed a survey. Results indicated both men and women reported that condoms hindered their sexual satisfaction, caused health problems for them and reduced their sexual interest. The findings also showed that obtaining condoms from clinics and perception that condoms do not cause health problems predicted the likelihood of condom use for both sexes. In addition, the results indicated that obtaining first time information from family members or relatives predicted the likelihood of condom use for women. It was recommended that intervention efforts should make condoms freely available in tertiary institutions, and such interventions should be tailored to overcome the relevant barriers that interfere with condom use for both the women and men. PMID- 16036232 TI - Exploring socio-economic conditions and poor follow-up rates of HIV-exposed infants in Johannesburg, South Africa. AB - In 2002, more than 280,000 HIV-exposed babies were born in South Africa. According to international PMTCT guidelines, these children require follow-up to 12 months of age. Worldwide, the high loss to follow-up rates experienced by PMTCT programs precludes them from identifying and managing HIV-infected children. Socio-economic factors have been identified as potential contributors to poor follow-up. A small descriptive study to examine socio-economic circumstances of women attending the Coronation Women and Children's Hospital PMTCT program was undertaken. Cross-sectional data from 176 women, interviewed at their infants' 12-month visit, was collected using a semi-structured questionnaire. Socio-economic factors such as poverty, geographical relocation and a lack of paternal support may affect the capacity of families to comply with the PMTCT follow-up program. Fifty-seven percent of mothers were unemployed, 25% of fathers did not support their children and only 58% of children remained resident in Johannesburg at the 12-month visit. The lack of follow-up of HIV infected children denies them access to adequate medical care. Understanding the socio-economic factors that affect the ability of communities to comply with PMTCT programs will assist resource-poor countries in devising strategies to achieve follow-up of HIV-exposed infants. PMID- 16036233 TI - The impact of social environments on the effectiveness of youth HIV prevention: a South African case study. AB - Few would disagree that 'social context' shapes the effectiveness of HIV prevention programmes. However much work remains to be done in developing systematic conceptualisations of HIV/AIDS-relevant aspects of social environments in vulnerable communities. This paper contributes to this challenge through a case study (44 interviews, 11 focus groups with 55 people and fieldworker diaries) of the impact of social context on a participatory peer education programme involving young people in a peri-urban community in South Africa. Three interacting dimensions of context undermine the likelihood of effective HIV prevention. Symbolic context includes stigma, the pathologisation of youth sexuality (especially that of girls) and negative images of young people. Organisational/network context includes patchy networking amongst NGOs, health, welfare and education representatives and local community leaders and groups. This is exacerbated by different understandings of the causes of HIV/AIDS and how to manage it. These challenges are exacerbated in a material-political context of poverty, unemployment and crime, coupled with the exclusion of young people from local and national decision-making and politics. HIV-prevention initiatives seeking to promote health-supporting social environments should work closely with social development programmes to promote young peoples' social and political participation, increase opportunities for their economic empowerment, challenge negative social representations of youth, and fight for greater recognition of their sexuality and their right to protect their sexual health. PMID- 16036234 TI - Social support networks and medical service use among HIV-positive injection drug users: implications to intervention. AB - The study used network analysis to identify forms and sources of social support associated with a medical services use among a medically underserved population living with HIV/AIDS. Participants were African American former or current injection drug users (n=295; 34% female, 45% current drug users and 17% AIDS diagnosed). Outcomes were access to the same medical provider, use of outpatient services and emergency room (ER) use with or without subsequent hospitalization. Controlling for AIDS diagnosis, insurance, current drug use and gender, access to the same medical care provider was associated with more females in one's support network and more network sources of emotional support, financial support and instrumental assistance. Adjusting for confounders, outpatient service use was associated with more female support network members and more sources of emotional support. Controlling for participants' drug use and insurance, sub-optimal emergency department use was associated with greater number of active drug users in one's support network. Contrary to other study findings, having a supportive sex partner was associated with lower access to medical care, and kin support was not associated with medical service use. Results indicate that specific sources and forms of social support had differential influences on the sample's utilization of medical services. The findings suggest that promoting HIV-positive African American injection drug users' support network functioning may help improve HIV medical services utilization among this medically underserved population. PMID- 16036235 TI - Prediction of adherence to antiretroviral therapy: a one-year longitudinal study. AB - The aim of this longitudinal study was to identify the determinants of adherence to antiretroviral therapy (ART) in HIV patients over a period of 12 months. A total of 376 individuals living with HIV treated with ART participated in the study. Data were collected at baseline and at three, six, nine and 12 months. Variables assessed were adherence, attitude, outcome expectancies, self-efficacy, patient satisfaction with the relationship with their physician, provision of social support, optimism, CD4 cell count, viral load and side effects. Predictors of adherence in the Generalized Estimated Equation (GEE) were: high perception of self-efficacy (OR=1.68; 95%CI 1.27-2.22), positive attitude towards taking medication (OR=1.56; 95%CI 1.18-2.06), not living alone (OR=1.47; 95%CI 1.04 2.08) and being a male (OR=2.81; 95%CI 1.47-5.34). Subsequent analysis showed that a positive attitude towards taking medication was associated with a high level of patient satisfaction with their physician, high perceived social support, being optimistic, living with HIV for five years or less and experiencing no side effects. Also, a strong sense of self-efficacy was associated with positive perception of social support, high level of patient satisfaction with their physician and not living alone. These results suggest that interventions aimed at improving adherence to ART should focus on reinforcing self-efficacy and developing a positive attitude towards taking medication. PMID- 16036236 TI - Quality of life, depression and fatigue among persons co-infected with HIV and hepatitis C: outcomes from a population-based cohort. AB - The objective of the study was to describe the additional burden generated by hepatitis C (HCV) infection among HIV-infected individuals as measured by self reported quality of life, depression and fatigue. The provincial HIV/AIDS Drug Treatment Program (DTP) distributes all antiretroviral medication in the province of British Columbia. Eligibility for accessing antiretrovirals is based on published guidelines commensurate with the International AIDS Society. Each participant is asked to complete a self-administered mailed questionnaire that includes patient sociodemographic information, quality of life measures (Medical Outcomes Study-Short Form (MOS-SF), mental health issues (Centre for Epidemiological Studies Depression scale (CESD) and fatigue information. HIV-HCV co-infected individuals were compared to HIV mono-infected individuals using parametric and nonparametric methods. Multivariate logistic regression was used to examine the impact of hepatitis C on quality of life, depression and fatigue, after controlling for sociodemographics and HIV-specific clinical characteristics. Of the 4,134 individuals who were sent a HIV/AIDS DTP survey in 1999, 2000 or 2001, 484 participants both returned one and had an HCV-antibody test result on file. Of the 484 participants eligible for this analysis, 105 (22%) were HCV-positive. In comparison to the 379 (78%) patients testing negative for HCV, a larger proportion of co-infected patients were female (18% versus 3%, p<0.001), aboriginal (20% versus 3%, p<0.001), had ever injected drugs (79% versus 5%, p<0.001), were unemployed (91% versus 49%, p<0.001) and lived in unstable housing (19% versus 1%, p<0.001) at the time they completed the survey. Co-infected patients reported more symptoms consistent with depression, increased fatigue and poorer quality of life. However, using multivariate modeling, it was determined that the impact of HCV on quality of life, depression and fatigue was better explained by the sociodemographic factors related to poverty and injection drug use, than by HCV itself. In conclusion, individuals co-infected with HIV and HCV represent a patient population with significant physical and mental health challenges. Although these patients experience poorer quality of life, increased depression and fatigue, this experience appears to be primarily related to socio economic issues rather than HCV infection. PMID- 16036237 TI - Medical and social differences between French and migrant patients consulting for the first time for HIV infection. AB - The objectives of the study were to describe the medical and social characteristics of patients consulting for the first time after diagnosis of HIV infection and to compare the medical and social characteristics between French and migrant patients. From 1 January 2001 to 31 December 2002, all consecutive adults consulting for the first time for HIV infection in two HIV outpatient clinics located in the western suburb of Paris, agreed to an interview based on an administered questionnaire regarding their medical and socioeconomic characteristics. Of the 203 patients (98 women, 105 men), one-third (n=70) was of French nationality. Delay (+/-SD) in access to HIV outpatient clinic after diagnosis was shorter in migrant than in French patients, respectively 7.6+/-29.6 months (median=0.5, range=0 to 196.6) and 23.8+/-51.4 months (median=0.9, range=0 to 199.7); p=0.005. There was no significant difference in the medical characteristics between the two groups of patients on their first consultation. However, most of the migrants were living in very poor socio-economic conditions with minimal resources thus sometimes delaying initiation of HAART. The development of social facilities for HIV-positive migrants should be a public health priority. PMID- 16036238 TI - Methamphetamine-dependent gay men's disclosure of their HIV status to sexual partners. AB - Disclosure of one's HIV status to a potential sexual partner has important HIV prevention implications. This paper qualitatively evaluates the social and sexual contexts that influence disclosure of HIV status among methamphetamine-dependent gay men enrolled in an outpatient drug treatment research program. As part of an open-ended, semi-structured interview, 34 HIV-positive and HIV-negative men discussed how, when, to whom and under what circumstances they reveal information about their HIV status. The four factors that influence participants' decision to disclose include: (1) an HIV-negative sexual partner's disclosure; (2) sexual venue (private versus public); (3) primary versus non-primary partner; and (4) the perceived risk of the sexual act. Sexual encounters among the men in this sample often occurred in public environments with non-primary partners, and involved use of illicit substances. In these social and sexual contexts, both HIV positive and HIV-negative participants believed that it is HIV-negative rather than HIV-positive men who should initiate safer sex dialogue and safer sex practices. Findings are helpful in crafting HIV-prevention interventions targeting substance-using gay men whose sexual practices place them at high-risk for HIV-infection. PMID- 16036240 TI - Determinants of HAART discontinuation among injection drug users. AB - The objective of this study was to identify psychosocial determinants of, and self-reported reasons for, HAART discontinuation among HIV-positive injection drug users (IDUs). We examined correlates between sociodemographic characteristics, drug use and risk behaviors, outcome expectations, adherence self-efficacy, social support and HAART discontinuation among 160 HIV-positive participants in the Vancouver Injection Drug Users' Study (VIDUS). Logistic regression was used to identify the factors independently associated with discontinuation of HAART. Seventy-one (44%) study participants discontinued HAART during the study period. Factors independently associated with discontinuation of HAART included recent incarceration (OR = 4.84, p = 0.022), negative outcome expectations (OR = 1.41, p = 0.001), adherence efficacy expectations (OR = 0.70, p = 0.003) and self-regulatory efficacy (OR = 0.86, p = 0.050). The most frequently cited reasons provided for discontinuing HAART were being in jail (44%) and medication side effects (41%). The results of this study suggest that psychological constructs derived from self-efficacy theory are highly germane to the understanding of HAART discontinuation behavior and interventions that may change it. Incarceration may result in interruptions in HAART among IDUs, and programmatic changes may be needed to promote optimal retention on HAART among incarcerated HIV-infected IDUs. PMID- 16036241 TI - Post-traumatic stress disorder among recently diagnosed patients with HIV/AIDS in South Africa. AB - This study examined the prevalence of and factors associated with post-traumatic stress disorder in recently diagnosed HIV/AIDS patients in South Africa. One hundred and forty-nine (44 male, 105 female) recently diagnosed HIV/AIDS patients (mean duration since diagnosis = 5.8 months, SD = 4.1) were evaluated. Subjects were assessed using the MINI International Neuropsychiatric Interview (MINI), the Carver Brief COPE coping scale and the Sheehan Disability Scale. In addition, previous exposures to trauma and past risk behaviours were assessed. Twenty-two patients (14.8%) met criteria for PTSD. Current psychiatric conditions more likely to be associated with PTSD included major depressive disorder (29% in PTSD patients versus 7% in non-PTSD patients, p = 0.004), suicidality (54% versus 11%, p = 0.001) and social anxiety disorder (40% versus 13%, p = 0.04). Further patients with PTSD reported significantly more work impairment and demonstrated a trend towards higher usage of alcohol as a means of coping. Discriminant function analysis indicated that female gender and a history of sexual violation in the past year were significantly associated with a diagnosis of PTSD. Patients whose PTSD was a direct result of an HIV/AIDS diagnosis (8/22) did not differ from other patients with PTSD on demographic or clinical features. In the South African context, PTSD is not an uncommon disorder in patients with HIV/AIDS. In some cases, PTSD is secondary to the diagnosis of HIV/AIDS but in most cases it is seen after other traumas, with sexual violation and intimate partner violence in women being particularly important. PMID- 16036242 TI - Barriers to accessing HIV/AIDS care in North Carolina: rural and urban differences. AB - Many HIV-positive individuals face multiple barriers to care and therefore frequently experience unmet medical and support services needs. Rural areas often lack the infrastructure to support the delivery of comprehensive HIV services; however, few studies have examined service barriers faced by rural residents with HIV/AIDS, particularly in the South where two-thirds of people living with HIV/AIDS in rural areas reside. We surveyed North Carolina HIV/AIDS case managers (N = 111) employed at state-certified agencies regarding barriers to medical and support services that influence medication adherence for their rural and urban living clients. For each of the seven barriers assessed (long travel for care, HIV-related stigma, and a lack of transportation; HIV-trained medical practitioners; housing; mental health services and substance abuse treatment), a substantial proportion of case managers (29-67%) reported it was a 'major problem'. For five of the seven barriers, rural case managers were significantly more likely to identify the barrier as a 'major problem'. Multivariate analysis revealed that rural case managers and case managers with more female clients reported a greater number of barriers. Because unmet medical and support service needs may result in poorer outcomes for HIV-positive individuals, barriers to these services must be identified and addressed, particularly in rural areas which may be highly underserved. PMID- 16036243 TI - Physical and sexual violence and health care utilization in HIV-infected persons with alcohol problems. AB - We examined interpersonal violence and its association with health care utilization and substance use severity among a cohort of 349 HIV-infected men and women with histories of alcohol problems assessed biannually up to 36 months. Data included demographics, lifetime interpersonal violence histories, age at first violence exposure, recent violence (prior six months), substance use severity and health care utilization (ambulatory visits, Emergency Department (ED) visits, hospitalizations) and adherence to HIV medication. Kaplan-Meier survival curves estimated the proportion of subjects experiencing recent violence. Generalized estimating equation regression models evaluated the relationship between recent violence, utilization and substance use severity over time, controlling for demographics, CD4 counts and depressive symptoms. Subject characteristics included: 79% male; mean age 41 years; 44% black, 33% white and 23% other. Eighty percent of subjects reported lifetime interpersonal violence: 40% physical violence alone, and 40% sexual violence with or without physical violence. First violence occurred prior to age 13 in 46%. Twenty-four (41%) of subjects reported recent violence by 24 and 36 months, respectively. In multivariate analyses, recent violence was associated with more ambulatory visits, ED visits and hospitalizations and worse substance use severity, but not medication adherence. Due to the high incidence and associated increased health care services utilization, violence prevention interventions should be considered for HIV-infected patients with a history of alcohol problems. PMID- 16036244 TI - An investigation into the health-related quality of life of individuals living with HIV who are receiving HAART. AB - The health authorities have recently accepted the routine provision of highly active antiretroviral therapy to persons living with AIDS in South Africa. There is a need to investigate the impact of HAART on the health-related quality of life of people living with HIV/AIDS (PLWHA) in a resource-poor environment, as this will have an influence on compliance and treatment outcome. The aim of this study was to explore whether HAART is efficacious in improving the self-reported health-related quality of life (HRQoL) in a group of PWLA in WHO Stages 3 and 4 living in a resource-poor community. A quasi-experimental, prospective repeated measures design was used to monitor the HRQoL over time in participants recruited to an existing HAART programme. The HRQoL of 117 participants was determined through the use of the Xhosa version of the EQ-5D and measurements were taken at baseline, one, six and 12 months. At the time of the 12-month questionnaire, 95 participants had been on HAART for 12 months. Not all participants attended all follow-up visits, but only two participants had withdrawn from the HAART programme, after two or three months. At baseline, the rank order of problems reported in all domains of the EQ-5D was significantly greater than at 12 months. The mean score on the global rating of health status increased significantly (p < 0.001) from a mean of 61.7 (SD = 22.7) at baseline to 76.1 at 12 months (SD = 18.5) It is concluded that, even in a resource-poor environment, HRQoL can be greatly improved by HAART, and that the possible side effects of the drugs seem to have a negligible impact on the wellbeing of the subjects. This bodes well for the anticipated roll-out of HAART within the public health sector in South Africa. PMID- 16036245 TI - Trends and predictors of HIV-positive community attachment among PLWHA. AB - Communities most affected by HIV/AIDS have been instrumental in shaping Australia's responses to the threat of the epidemic. There are recent signs that levels of engagement in communities based around HIV-positivity have changed: a diminished sense of an AIDS crisis, the relative success of highly active antiretroviral therapy (HAART), and an increasing individualization of the HIV experience may be contributing to changes in the way HIV-community is experienced. In this paper, we explore levels of engagement in HIV-positive community among a cohort of people living with HIV/AIDS (PLWHA) and seek to explain why some PLWHA engage in an HIV-positive community while others do not. Using multivariate logistic regression, we found that three factors were independently related to feeling part of an HIV-positive community: having been diagnosed with HIV prior to the advent of HAART; having more recently taken Bactrim or Septrin for PCP; and finding it easier to take 'pills' on time. Taken together, these results suggest that both historical effects, such as the introduction of HAART, and effects related to living with HIV, such as the experience of an AIDS-related illness, help explain HIV-positive community engagement among PLWHA. PMID- 16036246 TI - Complementary and alternative medicine and adherence to care among HIV-positive Latino gay and bisexual men. AB - The purpose of this study was to examine the degree to which HIV-positive Latino gay and bisexual men utilized complementary and alternative medicine (CAM), and to explore the relationship between CAM use and adherence to treatment. The sample consisted of 152 HIV-positive Latino gay and bisexual men. Eighty percent of participants reported use of some form of CAM. Asian CAM was most commonly used, followed by herbal remedies and Latino CAM. Adherence to Western medical care was also explored, with more than two-thirds of the sample adhering completely to recommendations concerning seeing the doctor, taking medications and following instructions. The relationships between CAM use and Western medical care were examined with logistic regressions. Those who used Latino CAM were less likely to keep doctors' appointments, follow physicians' recommendations and adhere to the prescribed medication regimen in the past three days. Plant-based CAM was also significantly related to non-adherence to Western medicine. PMID- 16036247 TI - Modeling censored-in-the-middle condom use data. AB - Many HIV prevention programs seek to reduce the risk of infection through increases in condom use. Condom use is often expressed as a proportion: the number of sexual contacts in which a condom is used divided by the total number of sexual contacts. The distribution of this proportion has several undesirable characteristics-the principal one is bimodality. Bimodality results from excessive numbers of 0% and 100% responses, creating distributions that are censored-in-the-middle. The purpose of this paper is to show how censored condom use data can be usefully modeled using Tobit regression. Tobit regression first transforms observed variable scores into latent variable scores, scores on an unobserved, hypothetical condom use variable, and then the latent variable scores are modeled using one or more explanatory variables. Data from the National Institute on Drug Abuse Cooperative Agreement for HIV/AIDS Community-Based Outreach/Intervention Research program for frequency of condom use and number of drug injecting sex partners were used to illustrate the method. We found that for every additional drug injecting sex partner, the probability of using condoms decreased by approximately 1%. PMID- 16036248 TI - Barriers to HIV/AIDS knowledge and prevention among deaf and hard of hearing people. AB - This study investigated knowledge about HIV/AIDS and barriers to HIV/AIDS education and prevention among deaf and hard of hearing people. Focus groups and individual interviews were used to elicit information from various groups of people with a hearing loss in different regions of New York State. Themes elicited in the interviews suggested that deaf sign language users are less knowledgeable about HIV/AIDS than oral deaf and hard of hearing participants, and that deaf adolescents have more knowledge than deaf adults. These findings likely reflect differences in levels of education and English proficiency. In addition, participants living in urban areas and in sizeable deaf communities are more exposed to information about HIV/AIDS than other participants. All participants reported difficulties in communication with medical providers, limiting their access to health information and proper medical care. Recommendations for the design and dissemination of educational materials and greater access to services for deaf and hard of hearing people are provided. PMID- 16036249 TI - The impact of substance dependence on HIV sexual risk-reduction among men with severe mental illness. AB - We explored the impact of substance dependence on the efficacy of an HIV sexual risk reduction intervention previously shown to be effective among men with severe mental illness by comparing rates of high-risk sexual behaviors among men with (n = 26) and without (n = 31) a lifetime history of substance dependence. We sub-divided subjects by alcohol and drug dependence status, comparing each intervention sub-group to the corresponding control sub-group. At each follow-up interval (six, 12 and 18 months), the intervention group as a whole and the non substance dependent participants showed a significant reduction in risk; the substance-dependent men showed no difference from controls. These data suggest that among men with severe mental illness, substance dependence may be a further impediment to HIV risk reduction. PMID- 16036250 TI - Factors associated with condom use among brothel-based female sex workers in Thailand. AB - This study sought to determine the actual levels of condom use by female sex workers (FSW) in Thailand brothels. Specifically, it examined the influence that the country of origin of the FSW patrons exerted on usage of a condom, and it assessed the relevancy of constructs from the Health Belief Model in predicting FSW requesting condom usage by their clients. A survey was administered to a cross-sectional non-randomized convenience sample of 150 FSW in four geographic locations in Thailand, which included: Bangkok, Chang Mai, Mae Hong Son and Other (comprised mainly of brothels in small villages). Statistical interpretation of the data indicated that Thai FSW requested condom use in 63% of the cases, while the overall mean of condom use was only 51%. Condom use by patrons' country of origin was as follows: westerners 76%, foreign Asians 52% and native Thai men 27%. These results fall considerably short of the Ministry of Health's goal of 100% condom use in Thai brothels. Furthermore, public health education initiatives need to effectively target the native Thai FSW patrons since they represent the clients least likely to use a condom (27%). PMID- 16036251 TI - Voluntary counseling and HIV testing for pregnant women in the Kassena-Nankana district of northern Ghana: is couple counseling the way forward? AB - This study reports the results of a cross-sectional questionnaire survey undertaken in the Kassena-Nankana district of Ghana to assess the perception and attitude of 270 antenatal clinic attendants towards voluntary counseling and HIV testing. It was found that although 92.6% (95% CI 88.8-95.4) of respondents indicated a willingness to get tested, only 51% (95% CI 45.0-57.2) considered HIV testing for pregnant women to be useful. Most (93.6%) indicated they would like their husbands (partners) to know the result of the test and 52.2% indicated that their husbands would be willing to accompany them to antenatal clinic (ANC) at least once during the pregnancy. The perception of the usefulness of HIV testing (OR = 8.5, 95% CI 1.8-40.0), the willingness to disclose test result to the husband (OR = 13.3, 95% CI 4.0-44.5) and perceived willingness of husband to accompany wife to antenatal clinic (OR = 5.2, 95% CI 1.4-19.8) were found to be independent predictors of a woman's willingness to get tested. The willingness to disclose test result to husband (OR 3.2, 95% CI 1.1-10.5) and knowledge of at least one mode of MTCT HIV transmission (OR = 2.1, 95% CI 1.2-3.6) were found to be independent predictors of a woman's perception that getting tested was useful. The results suggest that for pregnant women in this district, the willingness to get tested for HIV does not equate with the perception of the test's usefulness, and that spouses are likely to exert strong influence on the attitude of pregnant women towards VCT. Couple counseling facilitated through couple-friendly ANC services should be explored as strategy for the intended VCT program in this district. PMID- 16036253 TI - An empirical test of the information, motivation and behavioral skills model of antiretroviral therapy adherence. AB - Nearly perfect adherence to demanding antiretroviral therapy (ART) is now recognized as essential for HIV-positive patients to realize its life sustaining benefits. Despite the dire consequences of non-adherence, a large number of patients do not follow their ART regimen. While many factors influence adherence, the literature is dominated by studies on only one or a small set of them. Multivariate, theory-based models of adherence behavior are of great interest. The current study tested one such model, the Information, Motivation and Behavioral Skills (IMB) model of ART adherence (Fisher et al., under review). A sample of HIV-positive patients on ART in clinical care in Puerto Rico (N=200) provided data on adherence-related information, motivation and behavioral skills as well as adherence behavior per se. Structural equation model tests used to assess the propositions of the IMB model of ART adherence provided support for the interrelations between the elements proposed by the model and extended previous work. Implications for future research and intervention development are discussed. PMID- 16036254 TI - Pathways to HIV testing among adults aged fifty and older with HIV/AIDS. AB - Older adults (age 50 +) are less likely to be tested for HIV and are diagnosed at a later disease stage than younger individuals. To examine the barriers and facilitating factors to testing in this age group, interview data from 35 older men and women who tested HIV positive at age 50 or older were analysed. Participants described a variety of pathways to testing, related to gender, sexual orientation, drug use, and era of the epidemic. Older gay and bisexual men described three trajectories: proactively seeking out testing, delaying testing due to fear and hopelessness, and denying exposure to HIV. Heterosexual drug users and their partners followed two trajectories, depending on the phase of the epidemic: (1) delay due to the lack of knowledge or perceived risk for infection, and (2) delay due to psychological barriers and drug use, despite recognizing their risk. Finally, heterosexual non-drug-users were unaware of their risk. Across risk groups, physical symptoms and encouragement from health care providers were the primary triggers that facilitated testing. The finding that risk perception was a necessary, but not sufficient, condition for undergoing HIV testing suggests that interventions need to reduce barriers and encourage earlier HIV testing among older adults, in addition to promoting risk awareness. PMID- 16036255 TI - HIV/AIDS in fishing communities: challenges to delivering antiretroviral therapy to vulnerable groups. AB - Fishing communities have been identified as among the highest-risk groups for HIV infection in countries with high overall rates of HIV/AIDS prevalence. Vulnerability to HIV/AIDS stems from, the time fishers and fish traders spend away from home, their access to cash income, their demographic profile, the ready availability of commercial sex in fishing ports and the sub-cultures of risk taking and hyper-masculinity in fishermen. The subordinate economic and social position of women in many fishing communities makes them even more vulnerable to infection. In this paper we review the available literature to assess the social, economic and cultural factors that shape many fisherfolks' life-styles and that make them both vulnerable to infection and difficult to reach with anti retroviral therapy and continued prevention efforts. We conclude from the available evidence that fisherfolk will be among those untouched by planned initiatives to increase access to anti-retroviral therapies in the coming years; a conclusion that might apply to other groups with similar socio-economic and sub cultural attributes, such as other seafarers, and migrant-workers including small scale miners, and construction workers. PMID- 16036256 TI - Costs of treatment of Swiss patients with HIV on antiretroviral therapy in hospital-based and general practice-based care: a prospective cohort study. AB - Some ambulant people with HIV are cared for primarily by their general practitioner and some in an outpatient clinic. Costs and patterns of care in these settings were studied in 65 such patients based in Zurich, from a limited societal perspective (excluding patient costs) based on medical resource use. Antiretroviral therapy (ART), other medications and patient variables were collected prospectively, and non-medication resources (professional time and investigations) and treatment history data were collected from medical records and by record linkage to the Swiss HIV Cohort Study database. Cost differences between the settings were estimated using multiple regression, controlling for differences in case-mix. ART comprised 80% of the total cost, non-medication costs 15% and non-ART medications 5%. Total costs were higher in the outpatient clinic (estimated additional cost after controlling for case-mix = 3489 Swiss Francs per year at 1999 prices, 95% confidence interval 742 to 6236, p=0.017). The difference was accounted for by higher ART costs in the outpatient clinic, not through a tendency to use more expensive drugs or higher doses but rather through the use of more drugs concurrently. Differences in ART prescribing patterns between the doctors in the outpatient clinic and the general practitioners were considerable and appear worthy of further investigation. PMID- 16036257 TI - Coping strategies of HIV patients with peripheral neuropathy. AB - The aim of this study was to evaluate the association between coping strategies and reports of with pain and distress in patients with HIV-related peripheral neuropathy. Seventy-eight HIV seropositive subjects completed the Coping Strategies Questionnaire (CSQ), a self-report measure that assesses seven factors, the Brief Pain Inventory (BPI), the Brief Symptom Inventory (BSI) and the Beck Depression Inventory (BDI). Bivariate correlations revealed that younger patients used more Praying-Hoping (r=-.23, p<.04) and Catastrophizing (r=-.30, p<.007). t-tests demonstrated that women used more Praying-Hoping (t(76) = 3.42, p<.01), while Hispanic and African American patients used more Praying-Hoping more than Caucasians (F (1,77) = 22.11, p=.0005). Catastrophizing significantly predicted higher scores on the BDI (t=2.968, p=.004), the Global Severity Index (GSI) of the Brief Symptom Inventory BSI (t=2.400, p=.02); and pain interference on the Brief Pain Inventory BPI (t=2.996, p=.004) controlling for age, gender, and ethnic background. These results demonstrate that coping strategies may differ according to age, gender, and ethnic background in an HIV population, and that Catastrophizing predicts distress and interference with functioning in this sample. PMID- 16036258 TI - Mother to child transmission of HIV in Vitoria, Brazil: factors associated with lack of HIV prevention. AB - Screening HIV infection in pregnancy provides an ideal opportunity to make an early diagnosis in women in order to provide treatment to reduce vertical transmission to the newborn. The objectives were to describe the profile of HIV infected pregnant women attending municipal hospitals in Vitoria and to identify the causes associated with the lack of HIV therapeutic prophylaxis. Descriptive analysis of antenatal and HIV surveillance data of pregnant women in Vitoria, Brazil from 1997 to 2001 was performed. HIV infection was reported in 208 pregnant women. Their median age was 23 years, antenatal care was reported in 96.2% and HIV infection was diagnosed during antenatal care in 60.1%. Zidovudine (ZDV) therapy was initiated in 88% of the cases, 57.9% of which started before 20 weeks of gestation. The remaining 12% of the total of pregnant women received no therapy. Factors associated with lack of mother to child transmission (MTCT) prevention were: test not available (14 cases), delay of test result (seven cases) and patient's refusal to take medication (four cases). After delivery, 8.2% of the newborn did not receive ZDV and 14 women breastfed. Among the 196 live births, 3.1% (95%CI 1.9-4.3) of MTCT was reported. These data from an area with comprehensive HIV and antenatal surveillance provide an opportunity to identify gaps in public health efforts to reduce MTCT. PMID- 16036259 TI - Condom use in African adolescents: the role of individual and group factors. AB - This study set out to assess the ability of the Theory of Planned Behaviour (TPB) to predict and explain condom use in a traditional African context and in particular to assess the relative contributions of individual and normative constructs. A questionnaire survey was administered to 152 young adults (48% male, 52% female) at two points in time. Key constructs contained within the TPB were measured at Time 1. A short follow-up set of items was administered one week later at Time 2, thereby allowing for the measurement of actual behaviour. The results not only provide strong support for the predictive power of the TPB, since 67% of the variance in intention was explained, but also highlight the extent to which sexual behaviour in a rural location is governed by family/social influences. Subjective norm not only proved to be one of the most significant elements of the TPB model, together with self-efficacy (neither attitude nor perceived control were significantly involved), but 'my family' emerged as the most significant other. As such, the findings offer empirical evidence to support interventions that penetrate community networks. PMID- 16036260 TI - Back to work: correlates of employment among persons receiving highly active antiretroviral therapy. AB - The aim of this study was to quantify the level of employment at one-year and to determine potential predictors of future employment among HIV-positive persons on highly active antiretroviral therapy (HAART) in the province of British Columbia. Of the 392 individuals that were initially unemployed at baseline 63 (16.1%) found a job over the subsequent year. Factors associated with becoming employed included a baseline income over 10,000 dollars, having long-term disability or unemployment insurance as an income source, having higher CD4 cell counts, and better physical, social, and role functioning. Factors negatively associated with finding employment included having provincial assistance as an income source and having ever been an injection drug user (IDU). In multivariate analyses, not using provincial assistance as a source of income (Odds Ratio [OR] = 7.39; 95% CI: 3.26-16.7; p < 0.001) and higher MOS-SF role functioning (OR = 1.12 per 10 point increment; 95% CI: 1.03-1.21; p = 0.005) were independent predictors of becoming employed. In conclusion, our study demonstrates that while significant advances have been made in the reduction of HIV-related mortality, the majority of HIV-infected individuals on adequate treatment are still unable to be gainfully employed. PMID- 16036261 TI - Outcomes of an inner-city HIV mental health programme integrated with primary care and emphasizing cultural responsiveness. AB - Effectiveness of HIV-related mental health practice in inner cities has not been adequately documented despite an urgent need for interventions for populations disproportionately affected by the epidemic. Practices must take into account and respond to cultural issues of people of colour and the needs of those infected through injection drug use or heterosexual sex with IDUs. We report the effectiveness of an HIV mental health programme integrated with primary care and emphasizing cultural responsiveness. A sample of 47 of 188 patients who received psychiatric and psychotherapy services was compared with other subjects. The 47 had an average of 8.34 encounters over an average of 36.23 weeks. Consumers of project services used mental health care at a higher rate than did comparison subjects. Regression analyses indicated that utilization was related to reduction in reported mental health problems, HIV-related physical symptoms, and use of alcohol and powdered cocaine, as well as to improvement in social functioning. Our clients, individuals often viewed as not likely to avail themselves of or benefit from mental health care, were highly motivated for self-improvement and, given access to competent, convenient and culturally respectful services, improved their well-being in significant domains. PMID- 16036262 TI - Private providers and HIV testing in Pune, India: challenges and opportunities. AB - We explored HIV testing practices of private medical providers in an urban Indian setting in Pune, western India. 215 private practitioners (PPs) and 36 persons-in charge of private laboratories were interviewed in separate surveys. 77% of PPs had prescribed HIV tests and 94% of laboratories had performed HIV tests, or collected samples for HIV testing. Among those providers who had prescribed/performed tests, practices which violated national policy guidelines were found to be common. 55% of PPs and 94% of laboratories had not prescribed/performed confirmatory HIV tests, 82% of PPs had conducted routine HIV screening tests, 53% of PPs and 47% of laboratories had never counselled patients before testing, and 39% of laboratories reported breaching confidentiality of test results. PPs' knowledge about HIV tests was also inadequate, with 28% of PPs who had prescribed HIV tests being unable to name the tests they had advised. Prolific HIV testing in the private medical sector is accompanied by inappropriate practices and inadequate knowledge, reflecting deficiencies in the implementation of policy guidelines. The perspectives and needs of private providers, the major source of health care in India, need to be acknowledged. Supportive and regulatory mechanisms can be used to involve private providers in the delivery of better HIV testing services. PMID- 16036263 TI - Knowledge and acceptability of HAART among TB patients in Durban, South Africa. AB - From October 2002 to February 2003, we conducted semi-structured interviews on knowledge of HIV, antiretroviral treatment, and willingness to participate in voluntary counselling and testing and HAART with 54 consenting patients attending a tuberculosis (TB) clinic in Durban, South Africa. 74% of patients interviewed reported not knowing anything about antiretroviral treatment (HAART). Knowledge of antiretroviral drugs (ARVs) was restricted to use in preventing mother to child HIV transmission (MTCT). 57.4% of the patients reported having an HIV test in the past, but less than 10% were aware of their current HIV status. Patients who did not know their current HIV status expressed fear and hesitation about testing HIV positive. However, 91% of the patients expressed willingness to participate in HIV treatment given the opportunity. The findings from this study indicate that knowledge of HIV treatment is limited. As access to HAART is expanded, information about HIV treatment options will need to be disseminated. TB centres may present unique opportunities for disseminating HIV prevention, care, and treatment options. PMID- 16036264 TI - Patients referred to an urban HIV clinic frequently fail to establish care: factors predicting failure. AB - To measure the success with which patients newly entering outpatient care establish regular care, and assess whether race/ethnicity was a predictive factor, we conducted a medical record review of new patients seen 20 April 1998 to 31 December 1998 at The Thomas Street Clinic, a county clinic for uninsured persons. Patients were considered 'not established' if they never saw a physician in the 6 months after intake (the 'initial period'), 'poorly established' if seen but a > 6-month gap in care began in the initial period, and 'established' if there were no such gaps. Of 404 patients, 11% were 'not established', 37% 'poorly established', and 53% 'established'. Injection drug use as HIV risk factor (IDU), admitted current alcohol and drug use, age < 35 years, and CD4 count > or = 200 cells/mm(3) were most common in the 'not established' group and least common in the 'established' group. In multivariate ordinal logistic regression, difficulty establishing care was associated with IDU, admitted current alcohol use, and admitted former drug use. Age > 35 years was protective. Half the indigent patients entering care in this single-site study fail to establish regular care. Substance use and younger age are predictors of failure to establish care. PMID- 16036266 TI - Offering telephone counseling to smokers using pharmacotherapy. AB - Whereas telephone-based counseling has been found to be effective in supporting smokers interested in quitting smoking, it is not known whether proactive efforts to reach smokers receiving cessation medications will enhance their likelihood of successful quitting. We had an opportunity to test, in a health plan setting, an offer of telephone-based counseling with smokers identified from health plan records as recently filling a prescription for nicotine replacement therapy or bupropion. After we removed 31 members determined to be ineligible, 1,329 were randomly allocated to receive an invitation either to telephone-based counseling (n = 663) or to a control group (n = 666). On average, 7 days (range = 3-15 days) elapsed from the day of the prescription fill until the Center for Health Promotion began calling to invite members to participate in telephone counseling. The Center for Health Promotion was able to reach 49% of those in the intervention group (323/663). Of these members, 118 (37%) declined any participation. Therefore, in response to the proactive contact, 63% (205/323) of those reached and 31% (205/663) of those eligible participated in some smoking cessation counseling. At the 3-month follow-up, we observed an increased quit rate (33.1% vs. 27.4%) among health plan members randomized to telephone-based smoking cessation counseling. The results varied by gender and amount smoked. In addition, the variables associated with quitting in a multivariate logistic regression model included older age and using more than 30 days of medication. PMID- 16036267 TI - The association between patient-reported receipt of tobacco intervention at a primary care visit and smokers' satisfaction with their health care. AB - U.S. Public Health Service (USPHS) clinical guidelines for tobacco treatment recommend that providers routinely counsel smokers using a five-step algorithm (5A's): ask about tobacco use, advise smokers to quit, assess interest in quitting, assist with treatment, and arrange follow-up. A potential barrier to compliance is providers' concern that addressing smoking might alienate smokers, especially those not ready to quit. A survey was mailed to 1,985 patients seen at one of eight Boston-area primary care practices from January 1 to March 31, 2003, and identified as smokers by chart review. The survey assessed respondents' receipt of the 5A's at their visit and their satisfaction with the provider's tobacco treatment and with their overall health care. We used multivariable logistic regression models to assess the association between satisfaction with care and patient-reported receipt of each 5A step, adjusted for age, sex, education, race, health status, smoking intensity, readiness to quit, and length of relationship with provider. Of 1,160 respondents (58% response rate), 765 reported that they smoked at the time of the visit. They reported high levels of satisfaction with their tobacco-related care and overall care. Patient-reported receipt of each 5A step was significantly associated with greater patient satisfaction with tobacco-related care and with overall health care, even after adjusting for a smoker's readiness to quit smoking. Satisfaction with overall health care increased as counseling intensity increased. Patient reports of smoking cessation interventions delivered during primary care practice are associated with greater patient satisfaction with their health care, even among smokers not ready to quit. Providers can follow USPHS guidelines with smokers without fear of alienating those not yet considering quitting. PMID- 16036268 TI - Measuring provider adherence to tobacco treatment guidelines: a comparison of electronic medical record review, patient survey, and provider survey. AB - An accurate method of measuring primary care providers' tobacco counseling actions is needed for monitoring adherence to clinical practice guidelines. We compared three methods of measuring providers' tobacco counseling practices: electronic medical record (EMR) review, patient survey, and provider survey. We mailed a survey to 1,613 smokers seen by 114 Boston-area primary care providers during a 2-month period to assess what tobacco counseling actions had occurred at the visit (N = 766; 47% response rate). Smokers' reports were compared with the EMR and with their providers' self-reported usual tobacco counseling practices, derived from a provider survey (N = 110; 96% response rate). Patients reported receiving each counseling action more frequently than providers documented it in the EMR. Agreement between the patient survey and the EMR was poor for all 5A steps (kappa statistic = 0.01-0.22). Providers reported that they often or always performed each 5A action at a higher rate than indicated by EMR or patient report. However, providers who said they often or always performed individual 5A steps did not have consistently higher mean rates of EMR documentation or patient report than those who said they performed the 5A's less frequently. Little agreement was found among the three methods of measuring primary care providers' tobacco counseling actions. Implementing an EMR does not necessarily improve providers' documentation of tobacco interventions, but EMR adaptations that would standardize provider documentation of tobacco counseling might make the EMR a more reliable tool for monitoring providers' delivery of tobacco treatment services. PMID- 16036269 TI - Improving clinic- and neighborhood-based smoking cessation services within federally qualified health centers serving low-income, minority neighborhoods. AB - Within federally qualified health centers serving low-income, African American audiences, participatory approaches to system changes were organized through multidisciplinary committees that (a) drew on evidence-based guidelines, (b) guided system changes including the requirement of documenting smoking status and readiness to quit in encounter forms, (c) tested and refined practice improvements prior to their general adoption, and (d) guided development of neighborhood-based resources and supports for smoking cessation that were linked to clinic-based services. Documentation of smoking status or readiness to quit increased from 2% of encounter forms in the first 3 months to 94.3% in the last 3 months of the 24-month program. This rate remained over 90% throughout the following year. Exit interviews also indicated increased key clinic-based services, including "explained importance of quitting" (to 78% and 82% of interview respondents in the two intervention clinics in year 2), "tell you that you should quit" (to 80% in each), "tell you about nicotine gum...or other medications" (to 69% and 58%), "offer to help you quit" (to 61% and 64%), and "tell you about programs or help in your neighborhood" (to 51% and 56%). These rates exceeded those in one comparison clinic and equaled those in a second that also had launched a smoking cessation initiative. From exit interviews, improvements in neighborhood resources and support (e.g., people and activities that encourage nonsmoking) also exceeded those in comparison clinics. Thus, participatory approaches to system changes and quality improvement can enhance clinic- and neighborhood-based smoking cessation services within health centers serving low-income, minority populations. PMID- 16036270 TI - Systems change to improve health and health care: lessons from addressing tobacco in managed care. PMID- 16036271 TI - An integrated computer-based system to support nicotine dependence treatment in primary care. AB - The purpose of this study was to develop, implement, and evaluate the feasibility of an integrated computer-based system for tobacco-user identification and smoking cessation intervention for primary care patients in a medically indigent, managed care population. Interactive voice response (IVR) technology was used to screen for tobacco use prior to scheduled primary care visits at two inner-city clinics. The IVR system placed calls to 2,039 patients scheduled for clinic visits, and 1,086 (53%) patients completed the automated tobacco-use question set. Current smokers were identified in 421 (39%) of the calls. Computer generated reminders for clinicians that incorporated information obtained from the automated calls were placed on all smokers' encounter forms. In a post-visit interview of 120 smokers, 58 participants (48%) reported that they discussed smoking cessation with their provider. Some 71% of participants agreed that use of the IVR system to obtain information was a "good way for patients to give information about their health to doctors." Automated capture of patient-reported data via IVR technology is a potentially useful strategy for tobacco-use screening in primary care. PMID- 16036272 TI - Simplicity matters: using system-level changes to encourage clinician intervention in helping tobacco users quit. PMID- 16036273 TI - Tobacco attitudes, practices, and behaviors: a survey of dentists participating in managed care. AB - Tobacco cessation knowledge, attitudes, and behaviors of dentists participating in a large national managed care dental plan were assessed using a mailed survey. The survey was administered to dentists recruited to participate in an evaluation of a CD-ROM and supportive electronic detailing to promote increased tobacco cessation activities. General dentists who met specific technological criteria, had an active E-mail account, and at least 200 adult patients were eligible to participate in this study. A total of 184 dentists, located in 29 states, agreed to participate. The survey instrument included questions that addressed (a) Ask, Advise, Assess, Assist, and Arrange behaviors, (b) self-efficacy including knowledge, confidence, and success regarding tobacco cessation, (c) success of various tobacco cessation strategies, (d) barriers to tobacco cessation, and (e) demographics including year of graduation, gender, and race/ethnicity. Self reported baseline tobacco intervention-related behaviors were low, with 28% of dentists reporting that they asked their patients about tobacco or recorded tobacco use in their patients' charts at least 41% of the time. For Advise behavior, approximately half of the dentists advised tobacco-using patients to quit at least 41% of the time. Although self-reported lack of knowledge was high, 71% of respondents indicated that their lack of knowledge was either not a barrier or a slight barrier to incorporating tobacco cessation into their practices. The survey revealed that dentists do not routinely incorporate tobacco cessation into their practices. Newer information-transfer technologies may serve as vehicles for increased smoking cessation activities by dentists. PMID- 16036276 TI - Sociometric status as a predictor of onset and progression in adolescent cigarette smoking. AB - The relation between sociometric status and adolescent cigarette smoking was examined in a 1-year longitudinal study. Students in 6th, 7th, and 8th grades (N=1,630) were asked to nominate peers in their grade at school whom they liked and peers whom they disliked. These nominations were used to classify participants into one of five sociometric categories (popular, rejected, controversial, neglected, and average). Participants also reported their lifetime cigarette use at two measurements in consecutive school years. The results indicated that rejected and controversial adolescents were more likely than average adolescents to (a) report lifetime smoking at time 1 (T1) and (b) report onset of smoking at time 2 (T2). However, among adolescents who had already tried cigarettes at T1, rejected and controversial youth were not at increased risk for progression in total lifetime cigarette smoking (i.e., higher levels of total lifetime cigarette use at T2). Thus the results confirm that controversial youth are similar to rejected youth in their risk for onset of cigarette smoking during adolescence. PMID- 16036275 TI - Nicotine addiction through a neurogenomic prism: ethics, public health, and smoking. AB - Studies are under way to examine the neurogenetic factors contributing to smoking behaviors. The combined approaches of genomics, molecular biology, neuroscience, and pharmacology are expected to fuel developments in pharmacogenetics, to create new genetic tests, and ultimately to provide the basis for innovative strategies for smoking cessation and prevention. The emergence of a neurogenomic understanding of nicotine addiction is likely to induce fundamental changes in popular, clinical, and public health views of smoking, which could significantly shape existing practices and policies to reduce tobacco use. Still a nascent area of research, nicotine addiction provides an excellent case study through which to anticipate key ethical and policy issues in both behavioral genetics and the neurogenomics of addictive behaviors. PMID- 16036277 TI - Initial evaluation of a real-world Internet smoking cessation system. AB - To significantly reduce smoking prevalence, treatments must balance reach, efficacy, and cost. The Internet can reach millions of smokers cost-effectively. Many cessation Web sites exist, but few have been evaluated. As a result, the potential impact of the Internet on smoking prevalence remains unknown. The present study reports the results, challenges, and limitations of a preliminary, large-scale evaluation of a broadly disseminated smoking cessation Web site used worldwide (QuitNet). Consecutive registrants (N=1,501) were surveyed 3 months after they registered on the Web site to assess 7-day point prevalence abstinence. Results must be interpreted cautiously because this is an uncontrolled study with a 25.6% response rate. Approximately 30% of those surveyed indicated they had already quit smoking at registration. Excluding these participants, an intention-to-treat analysis yielded 7% point prevalence abstinence (for the responders only, abstinence was 30%). A range of plausible cessation outcomes (9.8%-13.1%) among various subgroups is presented to illustrate the strengths and limitations of conducting Web-based evaluations, and the tensions between clinical and dissemination research methods. Process-to outcome analyses indicated that sustained use of QuitNet, especially the use of social support, was associated with more than three times greater point prevalence abstinence and more than four times greater continuous abstinence. Despite its limitations, the present study provides useful information about the potential efficacy, challenging design and methodological issues, process-to outcome mechanisms of action, and potential public health impact of Internet based behavior change programs for smoking cessation. PMID- 16036278 TI - Acceptance of nicotine dependence treatment among currently depressed smokers. AB - This study reports on baseline characteristics associated with acceptance and refusal of available smoking treatment among currently depressed smokers in a psychiatric outpatient clinic who were enrolled in a larger clinical trial. The sample (N=154) was 68% female and 72% White, with a mean age of 41.4 years and average smoking rate of 17 cigarettes/day. All participants were assigned to a repeated contact experimental condition; received a stage-based expert system program to facilitate treatment acceptance; and were then offered smoking treatment, consisting of behavioral counseling, nicotine patch, and bupropion. Acceptors (n=53) were defined as those accepting behavioral counseling and pharmacological treatment at some point during the 18-month study, whereas refusers (n=101) received only the expert system. The number of days to treatment acceptance was significantly predicted by stage of change, with those in preparation entering treatment more quickly than contemplators or precontemplators. In a logistic regression, the variables most strongly associated with accepting treatment were current use of psychiatric medication and perceived success for quitting. Severity of depressive symptoms, duration of depression history, and history of recurrent depression were not related to treatment acceptance. Findings have implications for the psychiatric assessment and treatment of smokers in clinical settings. Psychiatric medication may play a significant role in smoking cessation treatment acceptance by currently depressed smokers. PMID- 16036279 TI - Tobacco education in U.S. schools of pharmacy. AB - This study is the first to characterize tobacco-related content in pharmacy school curricula in the United States. A national survey mailed to 83 U.S. schools of pharmacy assessed the extent to which tobacco is addressed in required coursework, educational methods of instruction, perceived importance of addressing tobacco in the doctor of pharmacy degree program, perceived adequacy of current levels of tobacco education in curricula, and perceived barriers to enhancing the tobacco-related content. A total of 82 surveys (98.8% response) revealed a median of 170 min of tobacco education throughout the doctor of pharmacy program. The most heavily emphasized topics are aids for cessation, assisting patients with quitting, nicotine pharmacology and principles of addiction, and drug interactions with smoking, yet more than 40% of respondents believed that each of these topics was covered inadequately. Key barriers to enhancing tobacco training are lack of curriculum time and lack of clinical clerkship sites focusing on tobacco interventions. Pharmacy faculty members perceive tobacco cessation training to be important, yet a mismatch exists between the perceived importance and the perceived adequacy of current levels of training in pharmacy school curricula. The results of this study will serve as a baseline measure against which future, parallel assessments will be compared as faculty at schools of pharmacy across the United States work together toward enhancing the tobacco cessation training of student pharmacists. PMID- 16036280 TI - Cigarettes, alcohol, and depression: characterizing head and neck cancer survivors in two systems of care. AB - Tobacco exposure is a key risk factor for head and neck cancer, and continued smoking after diagnosis negatively affects outcomes. The present study examined tobacco smoking, nicotine dependence, alcohol use, and depression in survivors of head and neck cancer. Subjects at least 6 months post-initial diagnosis of head and neck cancer (N=694) drawn from three VA otolaryngology clinics (n=309, VA patients) and a university-based otolaryngology clinic (n=385, non-VA patients) were administered questionnaires and standardized rating instruments for nicotine and alcohol dependence and for depression. Additional clinical information was extracted from chart reviews. Despite high rates of prior smoking, less than one quarter of all subjects continued to smoke. After controlling for significant confounding variables, we found that VA patients were more likely to be current smokers (OR=1.9, 95% CI=1.3-3.0), but current VA smokers did not differ significantly from non-VA smokers on the Fagerstrom Test for Nicotine Dependence criterion (p=.69). The VA patients were more likely to screen positive for problem drinking on the Alcohol Use Disorder Identification Test (OR=2.1, 95% CI=1.3-3.7). After adjusting for other variables, we found no statistical difference between the groups in depression scores on the Geriatric Depression Scale-Short Form. The study provides data on smoking, alcohol use, and depression in head and neck cancer survivors indicating that VA patients are at increased risk for continued smoking and problem drinking relative to non-VA patients. Head and neck cancer patients benefit from aggressive smoking cessation efforts by the VA, but many patients need specialized services that integrate smoking interventions with treatment of comorbid alcoholism. PMID- 16036281 TI - The influence of depressive symptoms on experimental smoking and intention to smoke in a diverse youth sample. AB - Numerous studies have shown associations between smoking and depression, but the generalizability of the relationship across ethnic groups remains unknown. The present study assessed the association between depression and smoking intention and experimentation among adolescents from four ethnic groups in the Los Angeles area-Chinese/Chinese American, Latino/Hispanic, Persian/Iranian, and White. Over 800 7th graders in the Los Angeles area completed measures of depressive symptoms, experimentation with smoking, intention to smoke, and sociodemographic covariates. Chinese/Chinese American students had the lowest levels of depressive symptoms, whereas Latinos/Hispanics had the highest levels. Latinos/Hispanics also were the most likely to intend to smoke in the next year and were the most likely to have started experimenting with cigarette smoking. Depressive symptoms were significantly associated with intention to smoke even after controlling for language use acculturation, socioeconomic status, gender, and ethnicity. The association between depressive symptoms and intention to smoke did not vary significantly across ethnic groups. These results indicate that the association between depressive symptoms and adolescent smoking generalizes across diverse ethnic groups. PMID- 16036282 TI - Preventing tobacco use among lesbian, gay, bisexual, and transgender youths. AB - A paucity of information regarding tobacco use among lesbian, gay, bisexual, and transgender (LGBT) youths impedes prevention programs. The aim of the present study was to conduct formative qualitative research regarding subpopulations at risk for tobacco use, protective factors, patterns of use, and approaches to prevention. This report focuses on participants' recommendations for the development of preventive intervention. Purposive sampling and maximum variation sampling were used to select 30 LGBT youths and 30 interactors for face-to-face interviews. NUD*IST6 text software was used for the indexing and thematic analysis of qualitative data, based on a grounded theory approach. All participants offered suggestions for tobacco prevention pertaining to the optimal process of prevention and cessation programs, specific strategies to promote tobacco prevention and cessation, and general strategies to foster nonsmoking. Several key themes regarding prevention emerged: LGBT youth should be involved in the design and implementation of interventions; prevention programs should support positive identity formation as well as nonsmoking; the general approach to prevention should be entertaining, supportive, and interactive; and the public might not distinguish primary prevention from cessation activities. All but one young smoker had attempted to quit at least once; but only one individual had succeeded. By way of implications, prevention programs should involve young people in enjoyable and engaging activities, address the psychosocial and cultural underpinnings of tobacco use, support healthy psychosocial development, and consider offering pharmacological smoking cessation aids. PMID- 16036283 TI - Recurrent event analysis of lapse and recovery in a smoking cessation clinical trial using bupropion. AB - We report a reanalysis of data from a prior study describing the event history of quitting smoking aided by bupropion, using recurrent-event models to determine the effect of the drug on occurrence of lapses and recoveries from lapse (resumption of abstinence). Data were collected on 1,070 subjects across two similar double-blind randomized clinical trials of bupropion versus placebo and fitted with separate Cox regression models for lapse and recovery. Analyses were split using discrete time-varying covariates between the treatment (weeks 1-10) and follow-up phases (end of treatment to 12 months). Bupropion was associated with slower lapse during treatment for both sexes, and being female was associated with faster lapse across both phases. Drug did not affect time to recovery for males but was associated with faster recovery among females, allowing women to recover as quickly as men. High levels of nicotine dependence did not affect time to lapse but were associated with slower recovery from lapse across treatment and follow-up phases. During the treatment phase, higher levels of baseline depression symptoms had no effect on time to lapse but were associated with slower recovery from lapse. Results highlight the asymmetry in factors preventing lapse versus promoting recovery. Specifically, dependence, depression symptoms, and a sex x drug interaction were found to affect recovery but not lapse. Further research disentangling lapse and recovery events from summary abstinence measures is needed to help us develop interventions that take advantage of bupropion at its best and that compensate where it is weak. PMID- 16036284 TI - A pilot study of smoking and associated behaviors of low-income expectant fathers. AB - Pregnancy is considered a teachable moment for helping women who smoke to quit, yet few studies have examined smoking behavior of expectant fathers. The present study considers the possibility that pregnancy is a teachable moment for expectant fathers as well and describes smoking and associated behaviors of men during their partner's pregnancy. Participants were 138 low-income men living with their pregnant partners. Using telephone interviews, we found 63% of the men had smoked at least 100 cigarettes in their lifetime. Current smoking was reported by 49.3% of expectant fathers (39.1% daily smoking; 10.2% some days). Expectant fathers' current smoking was associated with having a lower level of education (p<.0001), pregnant partner being a current smoker (p=.0002), higher quantity of alcohol consumption per day of drinking (p=.0003), and absence of smoking prohibitions inside the home (p<.0001). In the past year, 70.1% of the current smokers tried to quit. We found high rates of smoking in low-income expectant fathers, and an expectant father's smoking during his partner's pregnancy was associated with his pregnant partner continuing to smoke. A majority of expectant fathers identified as current smokers tried to quit in the past year or indicated an intention to quit in the near future. Intervention during pregnancy that targets pregnant women and expectant fathers who smoke could lead to more households without tobacco use and thus have positive implications for paternal, maternal, and family health. Further clinical and research attention is needed to address the smoking behaviors of both expectant fathers and their pregnant partners. PMID- 16036285 TI - Predictors of smoking reduction without formal intervention. AB - The Reduction of Smoking in Cardiac Patients (ROSCAP) Study is a randomized controlled trial to test the effectiveness of a smoking reduction strategy to decrease tobacco-related harm, promote cessation, and improve biochemical and clinical indicators of toxin exposure among patients with heart disease. We analyzed baseline characteristics of patients enrolled for participation to investigate predictors of spontaneous smoking reduction prior to study enrollment. Past reducers were more likely to be males (p=.009) and had higher past peak smoking level (p<.0001) than nonreducers. Gender and number of heart disease diagnoses predicted the occurrence of spontaneous reduction and its extent. Age and a history of chronic obstructive pulmonary disease also predicted the extent of reduction. Cotinine and nicotine levels per cigarette per day were significantly higher among past spontaneous reducers than nonreducers. Spontaneous reduction is common among medically ill smokers. Past history of heart disease is a strong predictor of prior reduction. There is evidence of compensation among past spontaneous reducers. PMID- 16036286 TI - Are Latinos really less likely to be smokers? Lessons from Oregon. AB - Our objective was to identify factors associated with current cigarette smoking among Latino adults in Oregon. We used data from 1,356 Latino participants and, for comparison, 18,593 non-Latino White participants in the 2000-2002 Oregon Behavioral Risk Factor Surveillance System (BRFSS). The BRFSS is a random-digit dialed, cross-sectional survey of noninstitutionalized, English- or Spanish speaking adults who live in Oregon households with a telephone. We examined relationships between current smoking and gender, age, education, and survey language (a measure of acculturation) among Latinos and used the .05 level of significance. The prevalence of current smoking was significantly lower among Latinos (18.1%) than non-Latino Whites (20.8%). Latino men were significantly more likely than Latina women to be smokers (23.1% vs. 11.4%), and some evidence indicated that less education was associated with smoking among Latino men. Taking the survey in English was strongly and significantly associated with smoking among Latina women: Smoking prevalence was 20.5% among those taking the survey in English and 3.3% for those taking it in Spanish. Our results suggest that the lower smoking prevalence among Latinos was driven by Latina women taking the survey in Spanish. Hence, it is essential for tobacco control programs to examine their local Latino smoking prevalence by gender and acculturation to avoid incorrectly concluding that Latinos are at decreased risk. Although programs for Latinos should reinforce protective aspects of Latino culture, decreasing the smoking prevalence in the U.S. mainstream culture might mitigate the negative impact of acculturation on smoking behaviors. PMID- 16036287 TI - Media advocacy and newspaper coverage of tobacco issues: a comparative analysis of 1 year's print news in the United States and Australia. AB - Tobacco control advocates now recognize the value of influencing news coverage of tobacco; news coverage influences attitudes and behavior as well as policy progression. It is, however, difficult to assess the progress of such efforts within a single national and temporal context. Our data represent the first systematic international comparison of press coverage of tobacco issues. Tobacco articles from major daily newspapers in Australia (12 newspapers; 1,188 articles) and the United States (30 newspapers; 1,317 articles) were collected over 1 year (2001). The analysis shows that coverage in the two countries was similarly apportioned between hard news (>70%) and opinion pieces. Similarly, stories in both countries were most likely to recount positive events. The substantive focus of coverage, however, differed, as did the expression of hostile opinion toward tobacco control efforts (United States, 4%; Australia, 7.1%). Although secondhand smoke and education, cessation, and prevention efforts were covered widely in both settings, these topics dominated coverage in Australia (29.2%) more than in the United States (17.6%), where a more diffuse set of tobacco topics gained relative prominence. The difference in policy conditions seems to offer contrasting opportunities for advocates in the two countries to use newspapers to promote helpful tobacco control messages for both behavior and policy change. PMID- 16036288 TI - Relationship between smokeless tobacco use and body weight in young adult military recruits. AB - Long-time cigarette smokers tend to weigh less than nonsmokers, and those who quit smoking typically gain weight. Little is known, however, about the relationship between smokeless tobacco and body weight. The present study investigated the association between smokeless tobacco use and body weight among 22,974 Air Force recruits (27.4% female, mean age=20.2 years, body mass index=22.7) undergoing basic military training. Current, former, and experimental smokeless tobacco users weighed significantly more than recruits who had never tried smokeless tobacco (p values <.05). Logistic regression analysis also indicated that the likelihood of being classified as overweight was significantly greater for daily (OR=1.29, 95% CI=1.07-1.54), occasional (OR=1.50, 95% CI=1.17 1.93), former (OR=1.33, 95% CI=1.05-1.67), and experimental (OR=1.13, 95% CI=1.02 1.24) smokeless tobacco users relative to never-users (p values <.05). These results suggest that smokeless tobacco use does not have significant weight attenuating effects, at least in the short term. Furthermore, using chewing tobacco or snuff may be associated with a greater body weight among young adults. PMID- 16036290 TI - Demographic analysis of post-abortive and interval-administered hormonal contraceptive methods. AB - OBJECTIVE: To compare, using method type and application time, the demographic characteristics of women using hormonal contraceptive methods administered after abortion or during the interval period with those of intrauterine device (IUD) users. METHODS: The demographic characteristics of women to whom oral contraception (OC), monthly injection, depot medroxyprogesterone acetate (DMPA), or IUD were administered in the post-abortive and interval periods between January 1998 and September 2001 in our clinic, were evaluated with the help of a registration system based on the Microsoft Access software used in our clinic. Demographic characteristics recorded were: age, reproductive expectation, previous contraceptive method, education level, number of live children, number of induced and spontaneous abortuses. RESULTS: A total of 10 500 women were included in the study. Of these, 6601 women had taken a contraceptive method during the interval period, and 3899 women had taken a contraceptive method post abortion. Only 4.6% of the cases were younger than 20 years, 48.3% were between 21 and 30 years, 37.2% were between 31 and 40 years and 9.9% were 41 years old or over. The DMPA-administered group contained the highest number of women > or = 41 years when compared to other groups (22.8%). In reproductive expectations, 65.8% of the cases wanted no more children; 1.3% wanted to have a baby within a 2-year period whereas 23.7% wanted a child after 2 years. The proportion of women wanting no more children was greatest in the DMPA-administered group (77.3%). No previous use of contraceptive methods was reported by 20.7% of women; 27% were using coitus interruptus. The education level of the women was as follows: 10% had no education, 58.3% had education to primary school level, 23.2% to secondary high-school level and 2.1% to university level. The group of women who chose OC as their contraceptive method contained the highest proportion of university graduates (6.5%). Only 6.26% of the women had no children. The proportion of women in the DMPA-use group with three or more children was higher compared to that in other groups (33.2%). CONCLUSION: The evaluation of demographic characteristics plays an important role in counseling, and in the efficacy and continued use of contraceptive methods. PMID- 16036291 TI - Ectopic pregnancy with postcoital contraception--a case report. AB - The incidence of ectopic pregnancies is increasing. Common risk factors are tubal pathology, previous tubal surgery, previous ectopic pregnancy, intrauterine device use and embryo transfer. Levonelle-2, a progesterone-only postcoital contraceptive works by a combination of mechanisms including ovulation inhibition, prevention of fertilization, and inhibition of implantation. It is 85% effective and there have been 12 reported cases of ectopic pregnancy in the UK with its use. It is believed that progesterone slows the intratubal migration of the fertilized ovum. In the case reported here, a woman presented with an ectopic pregnancy after use of Levonelle-2 as postcoital contraception; there were no clinical predisposing risk factors. In the absence of any histological evidence of tubal damage, we suspect that the levonorgestrel from Levonelle-2 could have been responsible for delayed embryo transfer which resulted in the ectopic pregnancy. PMID- 16036292 TI - Depot-medroxyprogesterone acetate in anticoagulated patients with previous hemorrhagic corpus luteum. AB - OBJECTIVE: To investigate the safety of depot-medroxyprogesterone acetate (DMPA) in women of reproductive age with prosthetic heart valves, as well as the impact of DMPA on the prevention of hemorrhagic corpus luteum in these patients with previous bleeding events. METHODS: In this prospective study we enrolled 13 patients who were receiving chronic anticoagulation for prosthetic heart valves, and who suffered from ovarian bleeding. After the initial bleeding episode(s), DMPA was initiated with the intent of preventing recurrent bleeding events by means of ovulation suppression. Follow-up included close monitoring of anticoagulation intensity, lipid profile, measurement of systolic and diastolic blood pressures and weight, and a general physical and gynecological examination. RESULTS: Of the participating 13 patients, one stopped DMPA after the third injection because she wanted to have a child. Among the remaining 12 women, over a mean follow-up of 39.9 months all patients were well and no hemorrhagic corpus luteum was observed. During the follow-up, anticoagulation intensity, assessed by the international normalized ratio (INR), was in the optimum therapeutic ranges at all times (range 2.5-3.5), except for values of 4.6, 5.8 and 5.9 in three patients at 9, 12 and 24 months, respectively. With regard to lipid profile, we observed a significant decrease in high-density lipoprotein cholesterol levels at 12 months, and significant increases in total cholesterol and triglyceride levels after 30 months compared to baseline serum levels. No significant changes were observed in serum low-density lipoprotein cholesterol levels. Mean body weight was higher at months 12 and 30, compared with baseline values (p > 0.05). CONCLUSION: DMPA, which is an effective contraceptive agent, can be used to prevent bleeding from the corpus luteum by means of ovulation suppression in anticoagulated patients with prosthetic heart valves. However, meticulous surveillance should be provided during the follow-up, including close monitoring of anticoagulation intensity and lipid profile. PMID- 16036293 TI - Pelvic abscess in intrauterine device users. AB - OBJECTIVE: To assess the causality between pelvic abscess formation and intrauterine device (IUD) use through a clinical study in a hospital. METHOD: Sixty-two pelvic abscesses were retrospectively evaluated over a 7-year period. Patient records retrieved for the women enrolled in this study consisted of demographic characteristics, duration of IUD use and clinical management details. RESULTS: All the women were monogamous Muslim women without any suspicious sexual contacts, immunosupressive states, or drug use at the time of IUD insertion. In 10 cases (16.1%), a history of pelvic surgery was present. The mean age of the women was 36.1 +/- 2.3 years (range 19-50 years). Of the 62 women, 14 (22.6%) were current IUD users. The mean time interval for women using IUD prior to the diagnosis of pelvic abscess was 5.7 +/- 1.2 years (range 1-14 years). In all cases, a pelvic mass and abdominal pain constituted the referral signs and symptoms. All women received an initial antibiotic regimen comprising penicillin (24 mU/day), clindamycin (900 mg/day) and gentamycin (240 mg/day) in divided doses. In 38 cases (61.3%), medical treatment yielded a satisfactory clinical outcome, defined as a decreas in mass volume together with pain relief and a decrease in leukocytosis. Twenty-four cases (38.7%) underwent a subsequent surgical procedure, either laparotomy (n = 19) or laparoscopy (n = 5). The type of surgery ranged from abscess drainage to more radical approaches such as total abdominal hysterectomy and/or unilateral or bilateral salpingo-oophorectomy. There were no differences between those women responding to medical therapy and those who did not respond in terms of mean age, percentage of past pelvic surgery, gravidity, parity and the size of pelvic abscess. CONCLUSIONS: A substantial number of women with an IUD were diagnosed as having a pelvic abscess within a 7-year period at the university clinic. Despite current knowledge that pelvic inflammatory disease and pelvic abscess are rarely encountered in long term IUD users, the presence of an IUD should be investigated in cases with an initial diagnosis of pelvic abscess based on clinical and ultrasonographic evaluation, demonstrating mostly acquisition via sexually transmitted disease. PMID- 16036294 TI - Contraception during perimenopause. AB - Perimenopause marks the transition from normal ovulation to anovulation and ultimately to permanent loss of ovarian function. Fecundity, the average monthly probability of conception, declines by half as early as the mid-forties, however women during the perimenopause still need effective contraception. Issues arising at this period such as menstrual cycle abnormalities, vasomotor instability, the need for osteoporosis and cardiovascular disease prevention, as well as the increased risk of gynecological cancer, should be taken into consideration before the initiation of a specific method of contraception. Various contraceptive options may be offered to perimenopausal women, including oral contraceptives, tubal ligation, intrauterine devices, barrier methods, hormonal injectables and implants. Recently, new methods of contraception have been introduced presenting high efficacy rates and minor side-effects, such as the monthly injectable system, the contraceptive vaginal ring and the transdermal contraceptive system. However, these new methods have to be further tested in perimenopausal women, and more definite data are required to confirm their advantages as effective contraceptive alternatives in this specific age group. The use of the various contraceptive methods during perimenopause holds special benefits and risks that should be carefully balanced, after a thorough consultation and according to each woman's contraceptive needs. PMID- 16036295 TI - Emergency contraception use by Irish teenagers. AB - OBJECTIVE: To audit teenagers attending a family planning clinic requesting emergency contraception. METHODS: A non-judgemental, relaxed, confidential interview was carried out. RESULTS: Many young Irish women become sexually active at a young age. Many teenagers appear to have problems using condoms correctly while others are taking chances by not using any method of contraception. CONCLUSIONS: These findings have important implications for those drawing up a sexual health strategy for young people. PMID- 16036296 TI - A rare case of ileal embedding by an intrauterine device. AB - Ileal penetration by a copper-bearing intrauterine device (IUD) is a rare but serious potential complication. This paper reports the case of a 30-year-old gravida 4 para 3 woman with ileal embedding by an IUD. Laparoscopy was performed on the patient 4 months after the insertion of the IUD. When the tail of the IUD was seen during laparoscopy protruding outside the small bowel, the device was removed through a 1-cm incision in the ileum by laparotomy. This report demonstrates a rare case of ileal embedding of an IUD. PMID- 16036297 TI - Antimicrobial peptides as microbicidal contraceptives: prophecies for prophylactics--a mini review. AB - The global increase in human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) and sexually transmitted infections (STIs) has led to the introduction of barrier methods, such as condoms. However, drawbacks associated with condoms, such as men being reluctant to use them and women being unable to negotiate their use, have led to the search for better and acceptable alternatives, namely the microbicides. These are gel formulations that, when used prior to sexual intercourse, protect against the transmission of HIV and other STIs. However, after observing the side-effects of nonoxynol-9, a component of the microbicidal formulations available on the market, the focus has shifted to natural available compounds demonstrating the preferred protective effects. Antimicrobial peptides (AMPs) are one such group of compounds present in a wide range of organisms from bacteria to humans. The existing 750 or so, low-molecular weight, cationic charged peptides are classified into five major groups based on their three-dimensional structure obtained by nuclear magnetic resonance studies. The hypothesized mode of action seems to be the interaction of the positively charged peptides with the negatively charged phospholipids present on the surface of the cell membrane. Various studies have demonstrated the effect of several AMPs, namely, defensins, protegrins, cathelicidins, cecropins, polyphemusins, magainins and melittins, against various STI-causing pathogens and HIV/herpes simplex virus, both in vitro and in vivo. The contraceptive efficacies of magainin and nisin in vitro and in vivo are worth mentioning. We believe these peptides are suitable candidates in the development of newer mechanism-based microbicides in future. PMID- 16036298 TI - Providers' knowledge of, attitude to and practice of emergency contraception. AB - OBJECTIVES: Barriers to widespread use of hormonal emergency contraceptives (EC), such as lack of knowledge and prejudices held by health-care providers, still exist today. This study was initiated to evaluate the knowledge, attitudes and prejudices of family-planning (FP) providers. METHODS: This survey was conducted in FP units of primary-health-care centers in Istanbul. A total of 180 providers were interviewed in 80 units to whom a questionnaire was administered by face-to face technique. RESULTS: One-hundred and fifty-two of the providers stated that they had heard of EC. The correct timing and dose interval of EC were known by 50% of them. The participants held the belief that EC caused abortion (39.4%), and that it was harmful for the fetus (31.1%). Other prejudices were the possibility of increased unprotected sexual intercourse (78.9%) and a tendency for men to give up condom use (75%); female providers were more prejudiced concerning these statements. The providers' tendency towards the provision of counseling was significantly related to their prejudices (p = 0.011, p = 0.033) and to the application rate (p = 0.000). Conclusion Providers need more detailed information about EC. During FP training courses, the providers should be encouraged towards counseling EC which would increase the application rate of the users and decrease their own prejudices. PMID- 16036299 TI - Accessibility and availability of abortion in six European countries. AB - OBJECTIVES: The accessibility and availability of abortion are a reflection of abortion law and the accessibility and availability of abortion services. Experiences from six European countries with different political, cultural, social and religious backgrounds (the Netherlands, France, Great Britain, Slovenia, Hungary and Turkey) are presented. RESULTS: Abortion laws in Europe range from complete prohibition to complete liberalization of abortion. Some countries demand a waiting period for the procedure, pre-abortion counseling, parental approval for minors, and in the others there are no mandatory requirements. Abortions are generally performed in authorized facilities by gynecologists or general practitioners. Abortion services are easily accessible, in terms of the law, availability of facilities and health insurance coverage of the procedure in the Netherlands, France and Slovenia. Abortion service is less accessible in United Kingdom, Hungary and Turkey, as a result of limited accessibility to abortion services or a relatively high abortion fee. In some Eastern European countries there has been a tendency in the last decade to limit the availability and access to abortion. CONCLUSIONS: In Europe abortion is generally well accessible in terms of abortion laws. There are differences in accessibility to abortion services between the countries, and in some countries, also between different areas of the country. PMID- 16036300 TI - Oral contraceptives use and weight gain in women with a Central European life style. AB - OBJECTIVES: The aim of the study was to check and present data on the relation between combined oral contraceptives (COC) use and body weight in young women living in Poland. METHODS: Observational study on the group of young women on COC and the control group of young women who never used COC. Candidates for study (145 women) and control (218 women) groups were recruited in selected gynecological clinics in Poland during the period of 1-31 January 2002 (with the use of randomization). RESULTS: Both groups (study and control) were almost identical in age, anthropometric characteristics, number of pregnancies and deliveries. On the basis of this investigation it was found that there was no relation between use of COC and weight gain. Observed (in both groups) weight gain in young women was most probably related rather to time-passing (patients were getting older). The higher risk for overweight and obesity was found in the group of young women who had already had problems with overweight in their childhood. and in the group of women with high weight gain during the first pregnancy. CONCLUSION: COC use is not associated with weight gain in young women with a Central European life-style. PMID- 16036301 TI - Use of a staff administered structured questionnaire to identify relevant life style issues and social-health determinants in a sexual and reproductive health service. AB - OBJECTIVE: To evaluate a staff-administered questionnaire to identify life-style issues and social-health determinants. DESIGN: Structured questionnaire administered by a health professional after dealing with the primary reason for attendance. SETTING: Community-based UK sexual and reproductive health service. POPULATION: First 1329 selected clinic patients comprising 1018 women attending Family Planning and 161 women plus 150 men attending Genitourinary Medicine; 47% were aged under 25. MAIN OUTCOME MEASURES: Identification of relevant health-risk taking, life-style issues, and unaddressed health concerns. Participants were offered appropriate support, information and referrals. RESULTS: Two hundred and sixty-four (23%) of the Family Planning women and 83 (52%) of the Genitourinary Medicine women [plus 103 (69%) of the men] reported two or more sexual partners in the last year. A third of participants denied regular condom use. Six per cent of women and 5% of men questioned had previously been forced to have sex. Eleven per cent of men admitted to having paid for sex and 9% of women disclosed physical assault (one-quarter in the home). Eight per cent of women and 7% of men had unresolved issues relating to previous miscarriage, termination, or stillbirth. CONCLUSIONS: It is possible to identify relevant life-style issues and social determinants of health during routine practice using a staff administered questionnaire. The resulting information may not otherwise have been disclosed and may impact significantly on health and care delivery. The information collected provides opportunities for both individuals and service planners to address wider health needs. PMID- 16036302 TI - Blood-brain barrier opening with alkylglycerols: Biodistribution of 1-O pentylglycerol after intravenous and intracarotid administration in rats. AB - Short-chain alkylglycerols have been described to increase the penetration of drugs and macromolecules across the blood-brain barrier (BBB) into the central nervous system (CNS) and were considered to be of potential value in the pharmaceutical treatment of CNS disorders. Due to the lack of information on the pharmacological behavior of these compounds in vivo, pharmacokinetics and biodistribution of [14C]- and [3H]-labeled 1-O-pentylglycerol (49 mg/kg, 100 mM) was investigated in normal male Wistar rats after intravenous as well as intracarotid administration. There was a rapid and predominant renal elimination of 1-O-pentylglycerol and more than 70% of administered dose was found in the urine within 270 min. Analysis of the pharmacokinetic parameters after a single i.v. bolus injection of 1-O-pentylglycerol resulted in a peak blood concentration of 0.58+/-0.06 micromol/ml, an initial half life of 23+/-7 min and a terminal half life of 18.8+/-4.1 h. No accumulation of 1-O-pentylglycerol was observed in the brain or other organs while highest concentrations were found in liver and thymus. This was confirmed by autoradiographic studies. Five minutes after intracarotid administration, high radioactivity was found in the ipsilateral brain, whereas after 30 min radioactivity in the brain has dramatically decreased. Autoradiographic images gave evidence of biliary excretion in addition to the renal elimination. There were no signs of cleavage of the O-alkyl bond in vivo as demonstrated by HPLC analysis. In conclusion, 1-O-pentylglycerol is characterized by pharmacological properties appearing very favorable for in vivo use as a permeabilizing drug for increased drug delivery to the brain. PMID- 16036303 TI - Preparation and characterization of a biodegradable drug targeting system for anticancer drug delivery: microsphere-antibody conjugate. AB - Targeted delivery of anticancer drugs is one of the most actively pursued goals in anticancer chemotherapy. A major disadvantage of anticancer drugs is their lack of selectivity for tumour tissue, which causes severe side effects and results in low cure rates. Any strategy by which a cytotoxic drug is targeted to the tumour, thus increasing the therapeutic index of the drug, is a way of improving cancer chemotherapy and minimizing systematic toxicity. This study covers the preparation of the gelatin microsphere (GM)-anti-bovine serum albumin (anti-BSA) conjugate for the development of a drug targeting approach for anticancer drug delivery. Microspheres of 5% (w/v) gelatin content were prepared by crosslinking with glutaraldehyde (GTA) at 0.05 and 0.50% (v/v) concentration. Microspheres were in the size range of 71-141?microm. The suitability of these microspheres as drug carriers for anticancer drug delivery was investigated in vitro by studying the release profiles of loaded methotrexate (MTX) and 5 fluorouracil (5-FU) and the cytotoxicities on cancer cell lines. The in vitro MTX release profiles (approximately 22-46% released in 24 h depending on the amount of GTA used) were much slower compared to 5-FU (approximately 42-91% released in 24 h). Both drugs demonstrated an initial fast release, which was followed by gradual, sustained drug release. The MTT cytotoxicity test results of GMs loaded with 5-FU and MTX showed approximately 54-70% and approximately 52-67% cytotoxicities in 4 days. In general, incorporation of MTX and 5-FU in microspheres enhanced the cytotoxic effect in a more prolonged manner compared to the free drugs. Gelatin micospheres were chemically conjugated to anti-BSA and the antigen-antibody activities were studied by immunofluorescence. Results indicated approximately 80% binding with conjugated anti-BSA and BSA-FITC. Based on their low cytotoxicity and the high antigen binding efficiencies, anti-BSA conjugated gelatin microspheres could be suitable targeted drug carrier systems for selective and long-term delivery of anticancer drugs to a specific body compartment (i.e. bladder cancer). PMID- 16036304 TI - Targeted delivery of arjunglucoside I using surface hydrophilic and hydrophobic nanocarriers to combat experimental leishmaniasis. AB - The purpose of the present study was to investigate the therapeutic efficacy of the indigenous drug arjunglucoside I (AG) against in vivo models of experimental leishmaniasis by incorporating it in surface hydrophilic co-polymeric nanogel particles of size less than 100 nm diameter and to compare its efficacy with that of the free drug as well as the drug encapsulated in hydrophobic poly-dl-lactide (PLA) nanoparticles. The drug AG, having glucose at the terminal end of the glycosidic chain, was isolated from an indigenous source. Drug-incorporated ultra low-sized nanogels (approximately 90 nm in diameter) composed of cross-linked random co-polymer of N-isopropylacrylamide (NIPAAM) and N-vinyl pyrrolidone(VP) were prepared, characterized and used as delivery vehicles to combat experimental leishmaniasis in hamster models. For comparison, drug-encapsulated hydrophobic nanoparticles (approximately 250 nm in diameter) made from PLA were used as a control. The drug AG was incorporated in these nanocarriers and these drug nanocarrier complexes were physically characterized. The efficacy of lowering spleen parasite load by the free drug, as well as that incorporated in nanogels and PLA nanoparticles were examined in vivo in equimolar concentration against hamsters undergoing experimental leishmaniasis. The reduction of drug toxicity by the nanogels and PLA nanoparticles was also assessed. The efficacy in the lowering of spleen parasite load with the free drug was found to be only 38% but was much higher when the drug was incorporated in co-polymeric nanogels (79%) or in polymeric nanoparticles (75%). Both the nanocarriers were found to be effective in reducing hepatotoxicity and nephrotoxicity nearly to the same extent. It was apparent that in addition to a smaller size and better drug release profile, the contribution of other parameters, e.g. overall surface hydrophilicity or hydrophobicity of the vehicles, also play an important role in the macrophage uptake of the drug. However, whatever be the exact mechanism, being highly efficient, non-hepatotoxic and non-nephrotoxic, AG in either of the two nanoparticulate forms may have useful application in humans PMID- 16036305 TI - Inverse targeting of diclofenac sodium to reticuloendothelial system-rich organs by sphere-in-oil-in-water (s/o/w) multiple emulsion containing poloxamer 403. AB - Sphere-in-oil-in-water (s/o/w) multiple emulsions containing diclofenac sodium were prepared by gelatinization of inner aqueous phase. A further modified version (s/o/wp) of s/o/w was formulated by adding 5.0% w/v poloxamer 403 to the external aqueous phase during the second step of emulsification in order to affect the adsorptive coating on the surface (s/o/wp). The inverse targeting of reticuloendothelial system (RES) rich organs was compared with a non reticuloendothelial system after intravenous administration of s/o/w multiple emulsion (treatment I) and poloxamer containing s/o/wp multiple emulsion (treatment II). The amount of diclofenac sodium in the plasma and various organs was measured to elucidate the effect of inverse targeting to RES and targeting to other tissues in terms of the incorporated drug. After i.v. administration, the half life (34.65 vs.16.26 h) and apparent volume of distribution of diclofenac sodium (2815 vs. 1671.5 ml/kg) were significantly higher in treatment II than in treatment I. It is concluded that the amount of drug in RES rich organs (spleen, liver) were significantly lower than the values in non-RES organs such as lungs, inflammatory tissue (synovial fluid) in treatment II than in treatment I. PMID- 16036306 TI - Polysorbate-stabilized solid lipid nanoparticles as colloidal carriers for intravenous targeting of drugs to the brain: comparison of plasma protein adsorption patterns. AB - Plasma proteins enriched on the surface of drug-delivery-purpose nanoparticles are regarded as key factors for determination of in vivo organ distribution after intravenous injection. Polysorbate 80-coated polybutylcyanoacrylate (PBCA) nanoparticles, preferentially adsorbing apolipoprotein E (apoE) on their surface, have previously been considered to deliver various drugs to the brain. In the present study, in vivo well tolerable solid lipid nanoparticles (SLN) using different types of polysorbates as stabilizers were produced. The influence of the different surfactants on in vitro adsorption of human plasma proteins was investigated using two-dimensional polyacrylamide gel electrophoresis (2-DE). Possible correlations of different amounts of adsorbed apoE to the hydrophilic lipophilic balance (HLB) of the polysorbates are shown and discussed. Apolipoprotein C-II, albumin and immunoglobulin G, which are also decisive plasma proteins with regard to site-specific drug delivery of intravenously injected carriers to the brain, are compared with regard to adsorption. Moreover, certain similarities to the plasma protein adsorption patterns of previously analysed brain-specific PBCA nanoparticles could be detected. Despite some differences in adsorption behavior of proteins on the surface of polysorbate-stabilized SLN and PBCA nanoparticles, we conclude that in both cases polysorbate 80 might have the highest potential to deliver drugs to the brain. PMID- 16036307 TI - Water-soluble polymers for targeted drug delivery to human squamous carcinoma of head and neck. AB - Human squamous cell carcinoma of the head and neck (SCCHN) is characterized by over expression of a tumor cell surface-specific receptor namely Hsp47/CBP2 that makes it a favorable candidate for targeted delivery of anticancer drugs. Several synthetic peptides have been identified as effective ligands for binding to CBP2. The purpose of this study is to investigate the potential of water-soluble N-(2 hydroxypropyl)methacrylamide (HPMA) copolymer-doxorubicin (Dox) conjugates containing a Hsp47/CBP2 binding peptide sequence, namely WHYPWFQNWAMA for targeted delivery to SCCHN. An HPMA copolymer containing Dox and CBP2 targeting peptide conjugated via lysosomally degradable glycylphenylalanylleucylglycine (GFLG) spacer was synthesized by free radical precipitation copolymerization. A control polymer without targeting moiety was also synthesized. The conjugates were characterized for drug content, peptide content, molecular weight and molecular weight distribution. The uptake of polymeric conjugates by both drug resistant and drug sensitive SCCHN cells were determined in vitro by flow cytometry using FACS scan analysis. Cytotoxicity of the conjugates towards drug sensitive as well as multidrug resistant SCCHN cells were evaluated by a clonal survival assay and compared to free Dox. The cytotoxicity of the free peptide was similarly evaluated. The internalization and subcellular fate of the conjugates in drug sensitive SCCHN cells was monitored using confocal microscopy. The new targetable copolymer contained 0.16 mmole peptide/g polymer. Studies on drug sensitive SCCHN cells demonstrated lesser uptake of both targeted and non targeted conjugates compared to free Dox suggesting a slower endocytic mechanism of uptake for the conjugates as opposed to rapid diffusion of free Dox. At higher Dox equivalent concentrations (>20 microM) the targeted conjugate showed significantly higher uptake (p < or = 0.028) than the non-targeted conjugate. The uptake of the targeted conjugate was inhibited in the presence of an anti Hsp47 antibody suggesting the involvement of active receptor mediated endocytosis in cell entry of the conjugate. Compared to free Dox, the targeted and non-targeted conjugates caused marginally lower inhibition (p < or = 0.01) of the drug sensitive SCCHN cells. In contrast, the same conjugates showed significantly higher uptake (p < or = 0.004) by drug resistant SCCHN cells and caused significantly higher inhibition (p < or = 0.02) of drug resistant SCCHN cells when compared to free Dox. Results suggest that the polymeric conjugates were able to overcome drug resistance. Confocal microscopy studies demonstrated the uptake of the polymeric conjugates, followed by internalization, intralysosomal localization and subsequent release of Dox. HPMA copolymer-Dox-peptide conjugates targeted to SCCHN cells were able to overcome drug resistance and increase efficacy in vitro. The results suggest that targetable polymeric conjugates have potential to improve systemic head and neck cancer chemotherapy by increasing tumor localization and reducing dose-limiting toxicity. PMID- 16036308 TI - Kinetic modelling of the intestinal transport of sarafloxacin. Studies in situ in rat and in vitro in Caco-2 cells. AB - The absorption kinetics of sarafloxacin, as a model of fluoroquinolone structure, were studied in the rat small intestine and in Caco-2 cells. The objective of the study was to investigate the mechanistic basis of the drug's intestinal transport in comparison with other members of the fluoroquinolone family and to apply a mathematical modelling approach to the transport process. In the rat small intestine, sarafloxacin showed dual mechanisms of intestinal absorption with a passive diffusional component and an absorptive carrier-mediated component. The characteristics of the animal study design made it suitable for population analysis, thus allowing the accurate estimation of transport parameters and their inter and intra-individual variances. The transport system in the rat model was ATP-dependent, as sodium azide was able to decrease the absorption rate constant in a concentration-dependent fashion. The inhibition mechanism of sodium azide was modelled based on its ATP depletion capacity. The rationale of this approach was to consider the inhibitor-carrier interaction as a concentration- dependent response. This interaction was accurately described by a non-competitive mechanism. In Caco-2 cells, sarafloxacin showed a concentration dependent permeability in both directions apical to basal, and basal to apical. The permeability values and ratios of permeability values at different concentrations suggested the presence of two carriers (absorption and efflux carriers). The passive diffusion component in both systems was compared to that predicted by the absorption-partition correlation, previously established for two series of fluoroquinolones. The discrepancy between the experimental and predicted value suggested the presence of an efflux mechanism similar to that already described for other fluoroquinolones. The differences and similarities of the in situ and the in vitro results are discussed as well as the usefulness of the modelling approach. PMID- 16036311 TI - Endothelium focus issue: molecular and cellular signaling in the perinatal cardiovascular system. PMID- 16036312 TI - The cerebral endothelium during pregnancy: a potential role in the development of eclampsia. AB - The authors investigated the influence of pregnancy on cerebral endothelial cell permeability in response to an acute elevation in intravascular pressure that caused forced dilatation of myogenic tone. Third-order branches of the posterior cerebral artery (PCA) were dissected from nonpregnant (NP) and late-pregnant (LP, days 19 to 20) Sprague-Dawley rats and mounted on glass cannulas in an arteriograph chamber that allowed control over intravascular pressure and measurement of both diameter and permeability to fluorescent dextran (3000 Da). Permeability was determined at 75 mm Hg and after a step increase in pressure to 200 mm Hg. The extent of pinocytosis and transcellular transport in response to pressure was evaluated separately in the same groups of animals at 75 and 200 mm Hg using transmission electron microscopy. All arteries developed myogenic tone at 75 mm Hg that was lost when pressure was increased to 200 mm Hg to cause forced dilatation. The increased pressure caused a significant increase in permeability to dextran and enhanced pinocytosis in arteries from LP animals, but not in NP animals whose permeability remained constant at both pressures. These results suggest a pregnancy-specific effect on the cerebral endothelium that may promote enhanced vascular permeability during acute hypertension and may contribute to the edema formation and neurologic complications of eclampsia. PMID- 16036313 TI - Functional significance of developmental changes in placental microvascular architecture. PMID- 16036314 TI - Effects of pulsatile shear stress on signaling mechanisms controlling nitric oxide production, endothelial nitric oxide synthase phosphorylation, and expression in ovine fetoplacental artery endothelial cells. AB - During gestation, placental blood flow, endothelial nitric oxide (NO) production, and endothelial cell nitric oxide synthase (eNOS) expression are elevated dramatically. Shear stress can induce flow-mediated vasodilation, endothelial NO production, and eNOS expression. Both the activity and expression of eNOS are closely regulated because it is the rate-limiting enzyme essential for NO synthesis. The authors adapted CELLMAX artificial capillary modules to study the effects of pulsatile flow/shear stress on ovine fetoplacental artery endothelial (OFPAE) cell NO production, eNOS expression, and eNOS phosphorylation. This model allows for the adaptation of endothelial cells to low physiological flow environments and thus prolonged shear stresses. The cells were grown to confluence at 3 dynes/cm2, then were exposed to 10, 15, or 25 dynes/cm2 for up to 24 h and NO production, eNOS mRNA, and eNOS protein expression were elevated by shear stress in a graded fashion (p < .05). Production of NO by OFPAE cells exposed to pulsatile shear stress was de novo; i.e., inhibited by L-NMMA (N(G) monomethyl-L-arginine) and reversed by excess NOS substrate L-arginine. Rises in NO production at 25 dynes/cm2 (8-fold) exceeded (p < .05) that seen for eNOS protein (3.6-fold) or eNOS mRNA (1.5-fold). Acute rises in NO production with shear stress occurred by eNOS activation, whereas prolonged NO rises were via elevations in both eNOS expression and enzyme activation. The authors therefore used Western analysis to investigate the signaling mechanisms underlying pulsatile shear stress-induced increases in eNOS phosphorylation and protein expression by "flow-adapted" OFPAE cells. Increasing shear stress from 3 to 15 dynes/cm2 very rapidly increased eNOS Ser1177, ERK1/2 (extracellular signal regulated kinase 1 and 2) and Akt, but not p38 MAPK (p38 mitogen-activated protein kinase) phosphorylation by Western analysis. Phosphorylation of eNOS Ser1177 under shear stress was elevated by 20 min, a response that was blocked by PI-3K (phosphatidylinositol 3-kinase) inhibitors wortmannin and LY294002, but not the MEK (MAPK kinase) inhibitor UO126. Basic fibroblast growth factor (bFGF) enhanced eNOS protein levels in static culture via a MEK-mediated mechanism, but it could not further augment the elevated eNOS protein levels induced by 15 dynes/cm2 shear stress. Blocking of either signaling pathways or p38 MAPK did not change the shear stress-induced increase in eNOS protein levels. Therefore, shear stress induced rapid eNOS phosphorylation on Ser1177 in OFPAE cells through a PI 3K-dependent pathway. The bFGF-induced rise in eNOS protein levels in static culture was much less than those observed under flow and was blocked by inhibiting MEK. Prolonged shear stress-stimulated increases in eNOS protein levels were not affected by inhibition of MEK- or PI-3K-mediated pathways. In conclusion, pulsatile shear stress greatly induces NO production by OFPAE cells through the mechanisms of both PI-3K-mediated eNOS activation and elevations in eNOS protein levels; bFGF does not further stimulate eNOS expression under flow condition. PMID- 16036315 TI - Pregnancy and ovarian steroid regulation of angiotensin II type 1 and type 2 receptor expression in ovine uterine artery endothelium and vascular smooth muscle. AB - Although pregnancy is clearly associated with refractoriness to infused angiotensin II (AII) in the uteroplacental unit, there is still dispute over the mechanism by which angiotensin type 1 and type 2 receptors (AT1R and AT2R) may mediate this response in the uterine artery. This is in large part due to incomplete knowledge of levels of AT1R and AT2R expression and function in uterine artery endothelium (UA Endo) in the nonpregnant (NP) and pregnant (P) states, combined with the disagreement on whether AII may act through release of adrenomedullary catecholamines. The authors have previously described an increase in AT1R in UA Endo but not UA vascular smooth muscle (VSM) during pregnancy as compared to the nonpregnant intact ewe. Herein they report that the pregnancy associated increase in AT(1)R expression in UA Endo is regulated by ovarian steroids. Using a recently developed antibody to AT2R, the authors now show there is no change in AT2R in UA Endo or VSM associated with ovarian function, and although AT2R is not changed in UA Endo by pregnancy, there is a significant decrease observed in UA VSM at that time. The authors also examined changes in receptors in UA Endo and VSM in estrogen (E2beta)-primed ewes in view of the common use of this model as a control for physiologic studies. In contrast to their findings in nonprimed nonpregnant or pregnant animals, the authors observed a significant increase in both AT1R and AT2R in UA Endo in response to the supraphysiologic priming with E2beta. In order to address the possible functionality of AT1R or AT2R in UA Endo, the authors used the uterine artery endothelial cell (UAEC) model of UA endothelial cells maintained in culture to passage 4. Differences in expression of AT1R or AT2R were normalized at passage 4 in P-UAECs and NP-UAECs. Treatment with AII activated phospholipase C (PLC) in both NP- and P-UAECs but signaling through the extracellular signal-regulated kinase (ERK) pathway was dramatically enhanced in P-UAECs compared to NP-UAECs. Surprisingly, both phosphoinositol turnover and ERK2 phosphorylation responses failed to display the expected dose-responses. Inhibition of AII-stimulated ERK2 phosphorylation with antagonists DUP 753 (AT1R, 10 microM) and PD 123319 (AT2R, 10 microM) failed to selectively inhibit ERK2 phosphorylation. The authors conclude that (a) the net effect of pregnancy may be an increase in the AT1R/AT2R ratio in both UA Endo and VSM but through apparently distinct mechanisms, (b) the ovariectomized animal model is similar to the luteal state for AT1R and AT2R expression, while the E2beta-primed model does not resemble the nonpregnant or pregnant state, and (c) there is a real possibility that AII may mediate its effects either through a complex AT1R-AT2R interaction or via an as-yet unidentified non-AT1, non-AT2 receptor. PMID- 16036316 TI - Mechanisms of renal vasodilation and hyperfiltration during pregnancy: current perspectives and potential implications for preeclampsia. AB - A thorough understanding of the renal and cardiovascular adaptations to normal gestation is essential for proper diagnosis and management of hypertensive disorders and renal diseases during pregnancy. Here, we briefly review the renal hemodynamic changes of normal pregnancy. In addition, we present new findings and current concepts related to the underlying hormonal and molecular mechanisms. Finally, we speculate on the potential contribution of these insights from normal pregnancy to the pathogenesis of preeclampsia. PMID- 16036317 TI - Intracellular signaling of lipid mediators via cognate nuclear G protein-coupled receptors. AB - Platelet-activating factor (PAF) and lysophosphatidic acid (LPA) are ubiquitous lipid mediators that play important roles in inflammation, cardiovascular homeostasis, and immunity and are also known to modulate gene expression of specific proinflammatory genes. The mechanism of action of these phospholipids is thought to be primarily dependent on their specific plasma membrane receptors belonging to the superfamily of G protein-coupled receptors (GPCRs). However, increasing evidence suggests the existence of a functional intracellular GPCR population. It has been suggested that immediate effects are mediated by cell surface receptors, whereas long-term responses are mediated by intracellular receptors. PAF and LPA(1) receptors localize at the cell nucleus of cerebral microvascular endothelial cells of newborn pig, rat hepatocytes, and cells overexpressing each receptor, and stimulation of isolated nuclei reveal biological functions, including transcriptional regulation of major genes, namely cylooxygenase-2 and inducible nitric oxide synthase. This mini review focuses on the nuclear localization and signaling of GPCRs, recognizing PAF and LPA phospholipids as ligands. Theories on how nuclear PAF and LPA1 receptors activate gene transcription and nuclear localization pathways are discussed. Intracrine signaling for lipid mediators uncover novel pathways to elicit their effects; moreover, intracellular GPCRs constitute a distinctive mode of action for gene regulation. PMID- 16036318 TI - Leptin, fetal nutrition, and long-term outcomes for adult hypertension. AB - One factor contributing to later hypertension, particularly in response to nutritional challenges is excess fat deposition around the kidney. In this review we discuss the hypothesis that these adverse conditions can be entrained by exposure of the conceptus to maternal nutrient restriction in early pregnancy. To this end we have shown in sheep that maternal nutrient restriction coincident with the time of embryogenesis and placental growth results in an early increase in fetal fat mass around the kidney that persists into later life. This is accompanied by an increase in leptin mRNA abundance and growth factor sensitivity. These adaptations occur in conjunction with reduced maternal plasma cortisol, thyroid hormones and leptin concentrations over the period of nutrient restriction. Some, but not all of these effects on fat development are accompanied by long term cardiovascular adaptations. As young adults, offspring from mothers nutrient restricted between early to mid gestation exhibit a leftward resetting, and blunting, of the cardiovascular baroreflex that appears to be mediated centrally through altered regional angiotensinogen II activity. At the same time, fat mass remains raised in nutrient restricted offspring. These animals demonstrate a marked increase in plasma leptin following sympathetic stimulation which is not observed in controls that indicates resetting of adipocyte sensitivity to stress. In conclusion, global nutrient restriction confined to the periods of embryonic and placental development therefore, programmes adult physiology, which may enhance predisposition to later disease given the appropriate environmental stimuli. PMID- 16036319 TI - Uterine venous permeability in the rat is altered in response to pregnancy, vascular endothelial growth factor, and venous constriction. AB - Venoarterial communication is a potential short-loop signaling pathway for the local control of uteroplacental perfusion. As this pathway is permeability dependent, this study investigated the effects of molecular weight, gestation, vascular endothelial growth factor (VEGF), wall tension, and constriction on solute flux across the venous wall. Experiments utilized fluorimetry to quantitate solute flux (3- and 70-kDa dextran) in isolated segments of uterine vein from virgin (NP) and late-pregnant (LP; day 20) rats as a function of endothelial surface area and time. Uterine veins were > 10-fold more permeable to the 3- versus 70-kDa dextran in both NP and LP groups. Flux was increased during gestation ( 2.5-fold), and by VEGF (NP, 3 kDa: 3.3-fold, 70 kDa: 4.8-fold; LP, 3 kDa: 3.3-fold, 70 kDa: 7.4-fold). Permeability to the 3 kDa dextran correlated directly with wall tension (r2 = .74 in both groups), whereas permeability to both dextrans correlated inversely with constriction (NP: r2 = .85 and .76; LP: r2 = .89 and .79, respectively). Uterine veins demonstrate permeability to 3- and 70-kDa tracers resulting from molecular weight dependence and an apparent difference in transport mechanisms. Permeability is enhanced by gestational remodeling and subject to modulation by placental factors, indicating the presence of a regulated pathway for the transfer of molecules across the venous wall. PMID- 16036320 TI - Antiangiogenic effect of soluble vascular endothelial growth factor receptor-1 in placental angiogenesis. AB - Differential splicing of the flt-1 mRNA generates soluble variant of vascular endothelial growth factor (VEGF) receptor-1 (sVEGFR-1, also known as sFlt-1). The action of VEGF is antagonized by sVEGFR-1. Soluble VEGFR-1 binds to VEGF with a high affinity and therefore works to modulate VEGF and VEGF signaling pathway. In this study, the authors tested the hypothesis that VEGF-mediated endothelial cell angiogenesis is tightly modulated by the release of sVEGFR-1 and placental expression of sVEGFR-1 is upregulated by hypoxia. Immunolocalization studies showed progressively intense staining for sVEGFR-1 and VEGF in the trophoblast of placental villous explants throughout gestation. Endothelial cell migration studies using a modified Boyden's chamber showed a significant increase in cell migration in response to VEGF that was significantly attenuated in the presence of exogenous sVEGFR-1. Furthermore, stimulation of endothelial cells with VEGF led to a dose-dependent increase in the release of sVEGFR-1 as determined by enzyme-linked immunosorbent assay (ELISA). Exposure of normal placental villous explants to hypoxia (1% pO2) increased trophoblast expression of sVEGFR-1 when compared with tissue normoxia (5% pO2). In addition, conditioned media from hypoxia treated placental villous explants induced a significant increase in endothelial cell migration that was significantly reduced in presence of sVEGFR 1. Our study demonstrates that hypoxia positively regulates sVEGFR-1 protein expression in ex vivo trophoblasts, which control VEGF-driven angiogenesis. PMID- 16036321 TI - In vitro biological properties of flavonoid conjugates found in vivo. AB - For some flavonoids such as quercetin, isoflavones and catechins, the pathways of absorption and metabolism are now reasonably well characterised and understood. By definition, for biological activity of flavonoids to be manifest, the target tissue, which includes the blood and vascular system, must respond to the form(s) of flavonoid that it encounters. Bioavailability studies have shown that the circulating form of most flavonoids is as conjugates, with a few notable exceptions. There have been several recent papers on the in vitro biological properties of conjugates that have been found in vivo. This paper reviews the properties of these conjugates. Most of the information currently available is on quercetin glucuronides, but also on isoflavone and catechin conjugates. In addition to the biological properties of the conjugates, the partition coefficients and methods of synthesis are also presented. PMID- 16036322 TI - Post-translational disulfide modifications in cell signaling--role of inter protein, intra-protein, S-glutathionyl, and S-cysteaminyl disulfide modifications in signal transmission. AB - Cell signaling entails a host of post-translational modifications of effector proteins. These modifications control signal transmission by regulating the activity, localization or half-life of the effector-protein. Prominent oxidative modifications induced by cell-signaling reactive oxygen species (ROS) are cysteinyl modifications such as S-nitrosylation, sulfenic acid and disulfide formation. Disulfides protect protein sulfhydryls against oxidative destruction and simultaneously influence cell signaling by engaging redox-regulatory sulfhydryls in effector-proteins. The types of disulfides implicated in signaling span (1) protein S-glutathionylation, e.g. as a novel mode of Ras activation through S-glutathionylation at Cys-118 in response to a hydrogen-peroxide burst, (2) intra-protein disulfides, e.g. in the regulation of the stability of the protein phosphatase Cdc25C by hydrogen-peroxide, (3) inter-protein disulfides, e.g. in the hydrogen peroxide-mediated inactivation of receptor protein-tyrosine phosphatase alpha (RPTPalpha) by dimerization and (4) protein S-cysteaminylation by cystamine. Cystamine is a byproduct of pantetheinase-catalyzed pantothenic acid recycling from pantetheine for biosynthesis of Coenzyme A (CoA), a ubiquitous and metabolically indispensable cofactor. Cystamine inactivates protein kinase C-epsilon (PKCepsilon), gamma-glutamylcysteine synthetase and tissue transglutaminase by S-cysteaminylation-triggered mechanisms. The importance of protein S-cysteaminylation in signal transmission in vivo is evident from the ability of cystamine administration to rescue the intestinal inflammatory-response deficit of pantetheinase knockout mice. These mice lack the predominant epithelial pantetheinase isoform and have sharply reduced levels of cystamine/cysteamine in epithelial tissues. In addition, intraperitoneal administration of cystamine significantly delays neurodegenerative pathogenesis in a Huntington's disease mouse model. Thus, cystamine may serve as a prototype for the development of novel therapeutics that target effector-proteins regulated by S-cysteaminylation. PMID- 16036323 TI - Endothelial NO Synthase (eNOS) phosphorylation regulates coronary diameter during ischemia-reperfusion in association with oxidative stress. AB - The link between endothelial nitric oxide synthase (eNOS) activation and vascular diameter during ischemia-reperfusion was investigated in the rat heart. After short (<30 min) and long (>45 min) time of ischemia conferred by coronary artery occlusion of the rats, reperfusion caused dilatation and constriction of arterioles, respectively. Partial oxygen pressure (pO2) measurement of the heart by the electrode confirmed the hyper-perfusion and no-reflow phenomena during reperfusion, as well as myocardial ischemia. The vascular diameter was correlated with phosphorylation of Akt and serine 1177 residue of eNOS, and formation of NO bound guanylate cyclase (GC) by immuoflorescence study. Western blotting confirmed the phosphorylation of eNOS-Ser1177 depending on ischemia time. The constriction during reperfusion after 45 min of ischemia is supposedly caused by the inhibition of Akt-mediated eNOS-Ser1177 phosphorylation, which was suppressed by a PKC inhibitor chelerythrine, or ROS scavengers N-2-mercaptopropionyl glycine (MPG) and 4,5-Dihydroxy-1, 3-benzenedisulfonic acid disodium salt (Tiron). However, an endothelin receptor antagonist BQ123 alleviated the vasoconstriction by increasing NO availability but not eNOS-Ser1177 phosphorylation. Thus, vascular patency is correlated with eNOS-Ser1177 phosphorylation in association with ROS, and PKC during reperfusion. Endothelin inhibits vasodilatation by reducing NO availability during reperfusion. PMID- 16036324 TI - Topical delivery of retinyl ascorbate: 4. Comparative anti-oxidant activity towards DPPH. AB - The free radical scavenging properties of retinyl ascorbate (RA-AsA) were determined by monitoring the decomposition of 2,2-diphenyl-1-picrylhydrazyl (DPPH) as a function of time and in comparison with ascorbic acid (AsA), ascorbic acid palmitate (AsA-Pal), retinoic acid (RA), retinol (ROL) and retinol palmitate (Rol-Pal). The rate constant of RA-AsA (mean3+/-SD) was 4.9+/-0.3 M(-1) s(-1), and indicated greater potency as an antioxidant compared to the rest of the test compounds (AsA 3.4+/-0.4 M(-1) s(-1), AsA-Pal, 2.9+/-0.2 M(-1) s(-1), RA 1.4+/ 0.3 M(-1) s(-1), ROL 1.3+/-0.1 M(-1) s(-1), Rol-Pal exhibited insignificant activity). The decomposition rate constant of DPPH, 5+/-0.6 x 10(-8) M(-1) s(-1), in ethanol and BHA, 154+/-3 M(-1) s(-1) were both used as control. The compound RA-2-carboxy-2-hydroxy-ethanoate was isolated by prep-TLC and was identified, by 13C and 1HNMR spectroscopy, as the major by-product from the reaction of RA-AsA with DPPH, which was also found to be potent antioxidant, 2.1+/-0.2 M(-1) s(-1). This suggests that oxidation of AsA moiety did not lead to the production of erythrulose species, which could cause deleterious modifications of cellular proteins. PMID- 16036325 TI - Differential DNA strand breaking abilities of *OH and ROS generating radiomimetic chemicals and gamma-rays: study of plasmid DNA, pMTa4, in vitro. AB - Reactive oxygen species (ROS) interact with components of a living cell. Among them *OH is known to cause major oxidative damage to living cells and is proposed to be involved in pathogenesis including carcinogenesis. Proper understanding of consequences of such damage is, therefore, medically relevant. In this report, aqueous solution of plasmid DNA, pMTa4, has been exposed to Fenton oxidant and Haber-Weiss oxidant mediated free radical generating chemical systems, and 60Co gamma-rays in vitro either alone or in combination to study their strand breaking abilities. The exposed pMTa4 was analyzed by agarose gel electrophoresis. The results show qualitative differences in the induction of strand breaks on the plasmid pMTa4 molecule by the iron (Fe2+), copper (Cu2+) or gamma-rays mediated *OH and other ROS. PMID- 16036326 TI - Acrolein induces Hsp72 via both PKCdelta/JNK and calcium signaling pathways in human umbilical vein endothelial cells. AB - Acrolein is a highly electrophilic alpha,beta-unsaturated aldehydes to which humans are exposed in a variety of environment situations and is also a product of lipid peroxidation. Increased levels of unsaturated aldehydes play an important role in the pathogenesis of a number of human diseases such as Alzheimer's disease, atherosclerosis and diabetes. A number of studies have reported that acrolein evokes downstream signaling via an elevation in cellular oxidative stress. Here, we report that low concentrations of acrolein induce Hsp72 in human umbilical vein endothelial cells (HUVEC) and that both the PKCdelta/JNK pathway and calcium pathway were involved in the induction. The findings confirm that the production of reactive oxygen species (ROS) is not directly involved in the pathway. The induction of Hsp72 was not observed in other cells such as smooth muscle cells (SMC) or COS-1 cells. The results suggest that HUVEC have a unique defense system against cell damage by acrolein in which Hsp72 is induced via activation of both the PKCd/JNK and the calcium pathway. PMID- 16036327 TI - Combination of glycemic and oxidative stress in lens: implications in augmentation of cataract formation in diabetes. AB - It is well known that the incidence of cataract is higher in diabetics as compared to non-diabetics. Its rate of maturation is also faster in the diabetics. The precise mechanism of this acceleration is not clearly understood. It is hypothesized that this could be a result of the combination of the metabolic and oxidative stress induced by glycemia itself with the age-associated increase in ambient generation of oxyradical species. In the current studies, we have investigated this possibility using the galactose cataract model. Galactosemia was induced by feeding rats a 50% galactose diet. The increased susceptibility of the glycemic lenses to physiological damage by reactive oxygen species (ROS) was studied by incubating them in Tyrode in the absence and presence of menadione. The resulting physiological damage to the lens was assessed initially in terms of its ability to maintain Na+-K+ ATPase dependent active transport of potassium ions, as represented by the uptake of rubidium ions. Subsequently, the level of ATP, indexing the general metabolic status, and the level of glutathione (GSH), indexing the status of antioxidant reserve, were also determined. The uptake of rubidium in the normal lenses incubated in the presence of the quinone was depressed to more than 50% of the controls run in the basal medium. A similar depression existed in the galactosemic lenses in comparison to the normal lenses. However, in the presence of menadione, the inhibition of the uptake was accentuated further in the case of galactosemic lenses, the uptake here being only 20% of the normal controls. Similarly, the galactosemic lenses were also more susceptible to menadione dependent decrease in ATP and GSH. PMID- 16036328 TI - Hydroperoxide characterisation as a signature of the micelle/monomer balance in radiation-induced peroxidation of arachidonate. AB - Archidonate peroxidation has been studied using HO* radicals radiolytically generated as initiators of this process. Irradiated aqueous solutions of arachidonate (between 0.01 and 25 mM at pH 10.5) have been characterised by means of conjugated dienes measurement (234 nm-absorption spectroscopy) and hydroperoxide detection (high-performance liquid chromatography coupled with a chemiluminescence detection). Radiation-induced peroxidation of arachidonate gives a different trend of peroxide products, depending on the degree of substrate interaction; endoperoxide and hydro-endoperoxide being favored at low concentrations (monomer/oligomer) and monohydroperoxide at high concentrations (micellar form). The experimental ratios G(Hydro2)/G(Hydro1) increase significantly only for arachidonate concentrations higher than 1 mM, i.e. in micellar medium. However, between 0.1 and 1?mM in arachidonate, G-values (for conjugated dienes, Hydro2 and Hydro1) remain nearly constant, meaning that the physical arrangement of the solution changes: Aggregation occurs. The experimental yields of conjugated dienes formation indicated that GDienes > GHO for [arachidonate]>2.5 mM, indicating that a chain propagation process had occurred. Radiolytic yields and structural identification (HPLC-MS analysis) of peroxidation products allowed us to propose a mechanism for the formation of both hydroperoxides. PMID- 16036329 TI - In vivo prooxidant state in Werner syndrome (WS): results from three WS patients and two WS heterozygotes. AB - The hypothesis was tested that Werner syndrome (WS) phenotype might be associated with an in vivo prooxidant state. A set of redox-related endpoints were measured in three WS patients, two of their parents, and 99 controls within a study of some cancer-prone and/or ageing-related genetic disorders. The following analytes were measured: (a) leukocyte 8-hydroxy-2'-deoxyguanosine; (b) glutathione from whole blood, and (c) plasma levels of glyoxal, methylglyoxal, 8-isoprostane, and some plasma antioxidants (uric acid, ascorbic acid, alpha- and gamma-tocopherol). Leukocyte 8-hydroxy-2'-deoxyguanosine levels showed a significant increase in the 3 WS patients vs. 85 controls (p<10(-7)). The disulfide glutathione:glutahione ratio was significantly altered in WS patients (p=0.005). Glyoxal and methylglyoxal levels were significantly increased (p=0.018 and p=0.007, respectively). The plasma levels of uric acid (p=0.002) and ascorbic acid (p=0.003) were also increased significantly in WS patients and in their parents. No significant alterations were found in the plasma levels of alpha- and gamma tocopherol, nor of 8-isoprostane. This is the first report of in vivo alterations of oxidative stress parameters in WS patients. Further investigations on more extensive study populations are warranted to verify the relevance of an in vivo prooxidant state in WS patients. PMID- 16036330 TI - Natural and newly synthesized hydroxy-1-aryl-isochromans: a class of potential antioxidants and radical scavengers. AB - We investigated the antioxidant and radical scavenging activity of polyphenolic isochromans. To assess the relation between structure and scavenging properties the natural occurring 1-(3'-methoxy-4'-hydroxy)phenyl-6,7-dihydroxy-isochroman (ISO-3, three OH groups) was compared with three newly synthesized derivatives that differ in their degree of hydroxylation by substitution with methoxy-groups (ISO-4: four OH groups; ISO-2: two OH groups and ISO-0: fully methoxylated). We found that ISO-4 is a 2-fold better scavenger for the artificial radical 1,1 diphenyl-2-picrylhydrazyl (DPPH, 100 microM) with an EC50=10.3 microM compared to the natural ISO-3 (EC50=22.4 microM) and to ISO-2 (EC50=25.1 microM), while ISO-0 did not react with DPPH. The scavenging capacity for superoxide enzymatically generated in a hypoxanthin-xanthinoxidase reaction was the highest for ISO-4 (EC50=34.3 microM) compared to those of ISO-3 (EC50=84.0 microM) and ISO-2 (EC50=91.8 microM), while ISO-0 was inactive. In analogy, ISO-4 scavenged peroxynitrite (ONOO-, EC25=23.0 microM) more effective than ISO-3, ISO-2 and ISO 0. When C6 rat glioma cells loaded with the reactive oxygen/nitrogen (ROS/RNS) sensitive fluorochrome 2,7-dichlorodihydrofluorescein, were exposed to hydrogen peroxide, the lowest stress level as indicated by the fluorescence signal was detected when the cells were pretreated with ISO-4 or ISO-2 but to a much lesser extent with ISO-3, while ISO-0 did not show any effect. All tested hydroxyisochromans superceded the scavenging effect of trolox.The excellent radical and ROS/RNS scavenging features of the hydroxy-1-aryl isochromans and their simple synthesis let these compounds appear to be interesting candidates for pharmaceutical interventions that protect against the deleterious action of ROS/RNS. PMID- 16036331 TI - Inhibition of the metmyoglobin-induced peroxidation of linoleic acid by dietary antioxidants: Action in the aqueous vs. lipid phase. AB - The gastric digestion of food containing oxidizable lipids and iron catalysts for peroxide decomposition such as (met)myoglobin from muscle meat can be accompanied by an extensive formation of potentially toxic lipid hydroperoxides. An early protective action by dietary antioxidants in the gastro-intestinal tract is plausible, especially for poorly bioavailable antioxidants such as polyphenols. Hence, the ability of antioxidants to inhibit lipid peroxidation initiated by dietary iron in mildly acidic emulsions is a valuable and general model. In this work, the ability of some ubiquitous dietary antioxidants representative of the main antioxidant classes (alpha-tocopherol, the flavonol quercetin, beta carotene) to inhibit the metmyoglobin-induced peroxidation of linoleic acid is investigated by UV-visible spectroscopy and HPLC in mildly acidic emulsions. The phenolic antioxidants quercetin and alpha-tocopherol come up as the most efficient peroxidation inhibitors. Inhibition by quercetin essentially proceeds in the aqueous phase via a fast reduction of an unidentified activated iron species (with a partially degraded heme) produced by reaction of metmyoglobin with the lipid hydroperoxides. This reaction is faster by, at least, a factor 40 than the reduction of ferrylmyoglobin (independently prepared by reacting metmyoglobin with hydrogen peroxide) by quercetin. By contrast, alpha-tocopherol mainly acts in the lipid phase by reducing the propagating lipid peroxyl radicals. The poorer inhibition afforded by beta-carotene may be related to both its slower reaction with the lipid peroxyl radicals and its competitive degradation by autoxidation and/or photo-oxidation. PMID- 16036332 TI - Effects of iron on Vitamin C/copper-induced hydroxyl radical generation in bicarbonate-rich water. AB - The aim of this study was to evaluate whether iron, like copper, could support Vitamin C mediated hydroxyl radical formation in bicarbonate-rich water. By using the hydroxyl radical indicator coumarin-3-carboxylic acid, we found that iron, in contrast to copper, was not capable to support Vitamin C induced hydroxyl radical formation. However, when 0.2 mg/l iron and 0.1 mg/l copper were both added to bicarbonate supplemented Milli-Q water, the Vitamin C induced formation of 7 hydroxycoumarin, as measured by HPLC analysis, was inhibited by 47.5%. The inhibition of hydroxyl radical formation by iron was also evident in the experiments performed on copper contaminated bicarbonate-rich household drinking water samples. In the presence of 0.2 mg/l of ferric iron the ascorbic acid induced hydroxyl radical formation was inhibited by 36.0-44.6%. This inhibition was even more significant, 47.0-59.2%, when 0.8 mg/l of ferric iron was present. None of the other redox-active metals, e.g. manganese, nickel or cobalt, could support ascorbic acid induced hydroxyl radical formation and did not have any impact on the ascorbic acid/copper-induced hydroxyl radical generation. Our results show, that iron cannot by itself produce hydroxyl radicals in bicarbonate rich water but can significantly reduce Vitamin C/copper-induced hydroxyl radical formation. These findings might partly explain the mechanism for the iron-induced protective effect on various copper related degenerative disorders that earlier has been observed in animal model systems. PMID- 16036334 TI - Regulation of protein function by glutathionylation. AB - The main function of reduced glutathione (GSH) is to protect from oxidative stress as a reactive oxygen scavenger. However, in the context of redox regulation, the ratio between GSH and its oxidized form (GSSG) determines the redox state of redox-sensitive cysteines in some proteins and, thus, acts as a signaling system. While GSH/GSSG can catalyze oxido-reduction of intra- and inter chain disulfides by thiol-disulfide exchange, this review focuses on the formation of mixed disulfides between glutathione and proteins, also known as glutathionylation. The review discusses the regulatory role of this post translational modification and the role of protein disulfide oxidoreductases (thioredoxin/thioredoxin reductase, glutaredoxin, protein disulfide isomerase) in the reversibility of this process. PMID- 16036335 TI - Effect of adriamycin treatment on the lifetime of pyrene butyric acid in single living cells. AB - We investigated the fluorescence lifetime of pyrene butyric acid (PBA) using various O2 concentrations in cells. Both in living and freshly fixed cells, PBA lifetime decreased with oxygen concentration. We recorded decay curves in single cells and measured PBA lifetime and NAD(P)H intensity values. Under nitrogen atmosphere, the probe lifetime differences (199 and 209 ns in living and freshly fixed cells, respectively) suggest a supplemental pathway for the deactivation of the probe when the cell functions are not stopped. We propose reactive oxygen species (ROS) to be the additional quenchers that cause this decrease. We further studied the effect of drugs generating ROS the anthracycline doxorubicin (adriamycin). For living cells, PBA lifetime decreased after adriamycin (ADR) treatment (200 and 1000 ng/ml). This supports our hypothesis that under nitrogen atmosphere and for freshly fixed cells, PBA lifetimes increase to an unchanging value due to absence of quenchers. PMID- 16036336 TI - Serum gamma-glutamyltransferase within its normal range predicts a chronic elevation of alanine aminotransferase: a four year follow-up study. AB - BACKGROUND: Previous epidemiological and experimental studies support the concept that serum gamma-glutamyltransferase (GGT) activity within its normal range is related to oxidative stress. Since oxidative stress plays a crucial role in the pathogenesis of various liver diseases, serum GGT may predict development of liver damage. METHODS: A total of 6,523 healthy male workers with normal alanine aminotransferase (ALT, <35 U/l) in a steel manufacturing company were followed for four years. Liver damage was defined as a chronic elevation of serum ALT (both 2001 and 2002). RESULTS: After adjusting for age, body mass index, alcohol consumption, cigarette smoking, exercise, and baseline value of ALT, in comparison with the group whose GGT level was <10 U/l, the adjusted relative risks for elevated ALT level among those with GGT levels 10-19, 20-29, 30-39, and over 40 U/l was 1.0, 2.5, 4.7, 7.4, and 12.0, respectively (P for trend <0.01). More importantly, this association was similarly observed even among non drinkers; the corresponding relative risks were 1.0, 1.8, 3.8, 5.6, and 6.2 (P for trend <0.01). However baseline ALT did not predict abnormal GGT level four years later. CONCLUSION: Serum GGT levels within normal range predict incidence of chronic elevation of ALT. Oxidative stress might explain this relationship. PMID- 16036337 TI - Oxidative DNA damage in cultured fibroblasts from patients with hereditary glutathione synthetase deficiency. AB - The SH compound glutathione (GSH) is involved in several fundamental functions in the cell, including protection against reactive oxygen species (ROS). Here, we studied the effect on oxidative DNA damage in cultured skin fibroblasts from patients with hereditary GSH synthetase deficiency. Our hypothesis was that GSH deficient cells are more prone to DNA damage than control cells. Single cell gel electrophoresis (the comet assay) in combination with the formamidopyrimidine DNA glycosylase enzyme, which recognizes oxidative base modifications, was used on cultured fibroblasts from 11 patients with GSH synthetase deficiency and five control subjects. Contrary to this hypothesis, we found no significant difference in background levels of DNA damage between cells from patients and control subjects. To study the induction of oxidative DNA damage without simultaneous DNA repair, the cells were gamma-irradiated on ice and DNA single-strand breaks measured. The patient and control cells were equally sensitive to induction of single strand breaks by gamma-irradiation. Therefore, factors other than GSH protect DNA from oxidative damage. However, cells with a high background level of oxidative DNA damage were found to be more sensitive to ionizing radiation. This suggests that differences in background levels of oxidative DNA damage may depend on the cells' intrinsic protection against induction of oxidative damage. PMID- 16036338 TI - Role of fibrillar Abeta25-35 in the inflammation induced rat model with respect to oxidative vulnerability. AB - The major pathological ramification of Alzheimer's disease (AD) is accumulation of beta-Amyloid (Abeta) peptides in the brain. An emerging therapeutic approach for AD is elimination of excessive Ass peptides and preventing its re accumulation. Immunization is the most effective strategy in removing preexisting cerebral Abetas and improving the cognitive capacity as shown in transgenic mice model of AD. However, active immunization is associated with adverse effect such as encephalitis with perivascular inflammation and hemorrhage. Details about the mechanistic aspects of propagation of these toxic effects are matter of intense enquiry as this knowledge is essential for the understanding of the AD pathophysiology. The present work aimed to study the oxidative vulnerability in the plasma, liver and brain of the inflammation-induced rats subjected to Ass immunization. Induction of inflammation was performed by subcutaneous injection of 0.5?ml of 2% silver nitrate. Our present result shows that the proinflammatory cytokines such as IL1alpha and TNFalpha are increased significantly in the inflammation-induced, Abeta1-42, Abeta25-35 treated groups and inflammation with Abeta25-35 treated group when compared to control, complete Freund's adjuvant and Abeta35-25 treated groups. These increased proinflammatory cytokines concurrently releases significant amount of free radicals in the astrocytes of respected groups. The present result shows that nitric oxide (NO) level was significantly higher (P<0.001) in plasma, liver and brain of the rat subjected to inflammation, Abeta1-42, Abeta25-35 and inflammation with Abeta25-35 injected groups when compared to control. The increased level of lipid peroxides (LPO) (P<0.001) and decreased antioxidant status (P<0.05) were observed in the plasma, liver and brain of inflammation-induced group when compared to control. Our result shows that significant oxidative vulnerability was observed in the inflammation with Ass treated rats when compared to other groups. Based on our results, we suggest that immunization of AD patients with Ass should be done with caution as the increase in Ass could trigger the brain inflammation in uncontrollable level. PMID- 16036339 TI - Melatonin, lipid peroxidation, and age in heterophils from the ring dove (Streptopelia risoria). AB - Numerous recent studies have shown the ability of physiological as well as all pharmacological concentrations of melatonin to prevent oxidative stress. We have found that incubating avian heterophils from young birds with a pharmacological concentration of 100 microM (23x10(6) pg/ml) melatonin reduced superoxide anion levels by modulating the activity of superoxide dismutase while also enhancing phagocytosis. There was also a decline in lipid peroxidation levels with both physiological and pharmacological concentrations of this indolamine. In the present work, we evaluated malonaldehyde (MDA) levels as an indicator of lipid peroxidation (both basal and antigen-induced) in young and old animals (ring doves) at different times of day (16:00 and 00:00) and with two incubation times (15 and 60 min). The lipid peroxidation was also measured in heterophils from old animals, incubated with the physiological concentrations of melatonin measured in young animals (50 and 300 pg/ml, diurnal and nocturnal, respectively). The results, expressed as nmol MDA/mg protein, show that MDA levels were higher in heterophils of old animals than in the young birds in all the experimental groups studied at both 16:00 and 00:00 (00:00 is the time at which the lowest peroxidation levels were obtained). Incubation with melatonin was found to reduce MDA levels, with the maximum reduction being after the 60?min incubation time and the nocturnal melatonin concentration. At both concentrations (diurnal and nocturnal), melatonin also counteracted the enhancement of MDA levels caused by latex beads, with the effect being greater at the longer incubation time. In conclusion, the results are further evidence of the antioxidant effect of melatonin even at physiological concentrations, and suggest its utility as a therapeutic agent in some pathological processes associated with age. PMID- 16036340 TI - Vitamin A deficiency induces prooxidant environment and inflammation in rat aorta. AB - We evaluated whether nutritional vitamin A deficiency generates oxidative stress and inflammation in aorta. Wistar male rats (21 days old) were given free access to a control (8 mg retinol as retinyl palmitate/kg) or a vitamin A- deficient diet for three months. One group of deficient animals was fed with the control diet fifteen days before sacrifice. Thiobarbituric acid-reactive substances (TBARS) and nitrite concentration where both analyzed in serum and aorta. Aorta Copper-Zinc Superoxide dismutase (CuZnSOD), Glutathion peroxidase (GPx) and Catalase (CAT) activities were measured. In addition, binding activity of the nuclear factor- kB (NF-kB), inducible and endothelial Nitric Oxide synthase (iNOS and eNOS, respectively) and Ciclooxygenase-2 (COX-2) expressions were determinated in aorta. Rats fed the vitamin A- deficient diet were characterized by sub-clinical plasma retinol concentration and showed increased serum and aorta concentrations of TBARS compared to controls. Lower than control activities of CuZnSOD, GPx, and CAT were observed in aorta of the vitamin A- deficient group. The binding activity of NF- kB was higher in vitamin A- deficient animals than controls. In addition, NO production evaluated as nitrite concentration increased in aorta and serum, associated with a higher expression of iNOS, eNOS and COX-2 in aorta of vitamin A-deficient rats. The incorporation of vitamin A into the diet of vitamin A-deficient rats reverted the changes observed in TBARS level, CuZnSOD and GPx activities, nitrite concentration and also, iNOS, eNOS and COX-2 expression. Prooxidant environment and inflammation are induced by vitamin A deficiency in rat aorta. PMID- 16036341 TI - Oxidative stress and inflammatory response during and following coronary interventions for acute myocardial infarction. AB - BACKGROUND: In acute myocardial infarction (AMI) treated with percutaneous coronary intervention (PCI), myocardial injury results from complex processes during both ischemia and reperfusion. Release of reactive oxygen species (ROS) may contribute to the accumulated myocardial damage. AIMS: To examine by frequent sampling of peripheral blood oxidative stress and early inflammation in patients undergoing primary PCI for AMI. Secondly, to assess whether a correlation exists between these parameters and the extent of myocardial damage. METHODS: Sixteen patients undergoing primary PCI within 6 h of AMI onset were included. Peripheral blood was sampled at start of procedure (t0) and repeatedly over 24 h following reperfusion. Main plasma analyses were: 8-iso-PGF2alpha (oxidative stress), 15 keto-dihydro-PGF2alpha (cyclooxygenase-mediated inflammation); and troponin-T (myocardial injury). Additional analyses included: total antioxidant status (TAS); vitamins; hsCRP and lipids. RESULTS: 8-Iso-PGF2alpha increased following restoration of blood flow, returned to t0 values after 3 h and was reduced below t0 the following day. TAS decreased significantly from t0 to the next day. There was no significant correlation between 8-iso-PGF2alpha and troponin T values. 15 Keto-dihydro-PGF2alpha was elevated during the first hour. There was a major rise in hsCRP after 24 h. CONCLUSION: Following reperfusion by primary PCI in AMI, oxidative stress and an inflammatory response are induced immediately. A rise in 8-iso-PGF2a during ischemia indicate that ROS generation may also take place during severely reduced coronary blood flow and hypoxia. No direct relationship between 8-iso-PGF2alpha or 15-keto-dihydro-PGF2alpha and troponin T was evident. The present study adds to the increasingly complex pathophysiological roles of ROS acting both as signal molecules and as mediators of tissue injury. PMID- 16036342 TI - Relationship among lipoperoxides, jasmonates and indole-3-acetic acid formation in potato tuber after wounding. AB - Plant responses to biotic and abiotic stress can be mediated by oxidised products and in this study we analysed the relation among some of them and the growth factor indole-3-acetic acid (IAA). The plant material used was potato tuber sliced below bud and incubated for different lengths of time before analysis. Wounding in potato tuber leads, in a very short time (0-30?min), to the generation of lipid hydroperoxides (LOOH) from polyunsaturated fatty acids (PUFA). These reactive species could cause a subsequent increase of 9 and 13 lipoxygenase (LOX, E.C.1.13.12.12.), analysed by RT-PCR and spectrophotometric assay, LOOH, Jasmonates and IAA all quantified by GC-MS analysis. The activation of 9 and 13-LOX, using different timing, leads to the formation of LOOH with a subsequent generation of jasmonates and IAA as highlighted by the addition on the potato tuber slices of salicylhydroxamic acid (SHAM), an inhibitor of LOX activity. A correlation between jasmonates and IAA resulted by testing their reciprocal influence during wounding in potato tuber. The relationship occurring among each hormone analysed during wounding underlines the fact that the jasmonates level can be regulated in situ and this can suggest a role for these compounds in potato tuber which has been underestimated up to now. PMID- 16036343 TI - Enhancement of quinone redox cycling by ascorbate induces a caspase-3 independent cell death in human leukaemia cells. An in vitro comparative study. AB - Since the higher redox potential of quinone molecules has been correlated with enhanced cellular deleterious effects, we studied the ability of the association of ascorbate with several quinones derivatives (having different redox potentials) to cause cell death in K562 human leukaemia cell line. The rationale is that the reduction of quinone by ascorbate should be dependent of the quinone half-redox potential thus determining if reactive oxygen species (ROS) are formed or not, leading ultimately to cell death or cell survival. Among different ROS that may be formed during redox cycling between ascorbate and the quinone, the use of different antioxidant compounds (mannitol, desferal, N-acetylcysteine, catalase and superoxide dismutase) led to support H2O2 as the main oxidizing agent. We observed that standard redox potentials, oxygen uptake, free ascorbyl radical formation and cell survival were linked. The oxidative stress induced by the mixture of ascorbate and the different quinones decreases cellular contents of ATP and GSH while caspase-3-like activity remains unchanged. Again, we observed that quinones having higher values of half-redox potential provoke a severe depletion of ATP and GSH when they were associated with ascorbate. Such a drop in ATP content may explain the lack of activation of caspase-3. In conclusion, our results indicate that the cytotoxicity of the association quinone/ascorbate on K562 cancer cells may be predicted on the basis of half redox potentials of quinones. PMID- 16036344 TI - Antioxidant supplementation reduces inter-individual variation in markers of oxidative damage. AB - The aim of this study was to examine the effect of antioxidant supplementation on oxidative damage and chromosome stability in middle-aged men, smokers and non smokers. A total of 124 men aged 48+/-6 years from Bratislava and from the rural population near Bratislava were investigated; 64 men (22 smokers and 42 non smokers) were supplemented for 12 weeks with antioxidants, while 60 (25 smokers and 35 non-smokers) were given placebo. The daily antioxidant supplementation consisted of vitamin C (100 mg), vitamin E (100 mg), ss-carotene (6 mg), and selenium (50 microg). Samples of blood were taken on two occasions: At the beginning and at the end of the supplementation trial. Concentrations of dietary antioxidants, ferric reducing ability, malondialdehyde as an indicator of lipid peroxidation in plasma, micronuclei and chromosome aberrations in lymphocytes were measured. Antioxidant supplementation significantly increased the levels of vitamin C, ss-carotene, a-tocopherol and selenium in plasma. The overall antioxidant status of plasma measured as ferric reducing ability of plasma (FRAP) increased significantly (p<0.001) after antioxidant supplementation as well. The increase in antioxidant parameters after supplementation were consistently more pronounced in non-smokers than in smokers. There was a significant decrease of malondialdehyde concentration in the non-smokers, while in smokers the decrease of malondialdehyde concentration was not significant. Antioxidant supplementation did not affect the proportion of lymphocytes with micronuclei or the total number of micronuclei; however, there was a significant positive correlation (p<0.001) between the malondialdehyde concentration at the beginning of the supplementation trial and the difference in number of cells with micronuclei before and after the supplementation. The percent of cells with chromosome aberrations decreased significantly after antioxidant supplementation in smokers. These results indicate that a combined antioxidant supplementation (a) is effective even at very moderate doses; (b) significantly diminishes oxidative damage to lipids when it is high initially; and (c) is effective in decreasing chromosomal instability in lymphocytes of middle-aged men. PMID- 16036346 TI - Review: When is an antioxidant not an antioxidant? A review of novel actions and reactions of vitamin C. AB - Vitamin C (or ascorbic acid) is regarded as the most important water-soluble antioxidant in human plasma and mammalian cells which have mechanisms to recycle and accumulate it against a concentration gradient, suggesting that the vitamin might also have important intracellular functions. In this review we summarize evidence from human trials that have attempted an association between vitamin C supplementation and an effect on biomarkers of oxidative DNA damage. Most studies reviewed herein showed either a vitamin C-mediated reduction in oxidative DNA damage or a null effect, whereas only a few studies showed an increase in specific base lesions. We also address the possible beneficial effects of vitamin C supplementation for the prevention of cancer and cardiovascular disease. Finally, we discuss the contribution of cell culture studies to our understanding of the mode of action of vitamin C and we review recent evidence that vitamin C is able to modulate gene expression and cellular function, with a particular interest in cell differentiation. PMID- 16036347 TI - Effect of auranofin on the mitochondrial generation of hydrogen peroxide. Role of thioredoxin reductase. AB - The mitochondrial production of hydrogen peroxide, in the presence of different respiratory substrates (succinate, glutamate, malate and isocitrate), is stimulated by submicromolar concentrations of auranofin, a highly specific inhibitor of thioredoxin reductase. This effect is particularly evident in the presence of antimycin. Auranofin was also able to unmask the production of hydrogen peroxide occurring in the presence of rotenone. However, at variance with whole mitochondria, auranofin does not stimulate hydrogen peroxide production in submitochondrial particles indicating that it does not alter the formation of hydrogen peroxide by the respiratory chain but prevents its removal. As the mitochondrial metabolism of hydrogen peroxide proceeds through the peroxidases linked to glutathione or thioredoxin, the relative efficiency of the two systems and the effects of auranofin were tested. In conclusion, the inhibition of thioredoxin reductase determines an increase of the basal flow of hydrogen peroxide leading to a more oxidized condition that alters the mitochondrial functions. PMID- 16036348 TI - Accelerated impairment of spermatogenic cells in SOD1-knockout mice under heat stress. AB - For normal spermatogenesis, the temperature of the scrotum is lower than that of the body. The mechanism by which mammalian testes undergoes cell death as the result of exposure to heat continues to be a matter of debate. Since generation of reactive oxygen species (ROS) during heat stress and involvement in spermatogenic cell damage are postulated, we induced experimental cryptorchidism in the testes of SOD1-knockout mice and examined effects of the gene deficiency. The cleavage of DNA in testicular cells, as judged by TUNEL staining, were elevated in SOD1-knockout mice at an earlier stage than in the wild-type mice. To confirm responsiveness of SOD1 for this high susceptibility to heat stress, spermatogenic cells were isolated from SOD1-knockout and wild-type mice and cultured at 32.5 and 37 degrees C. The cells isolated from SOD1-knockout were more vulnerable at both temperatures than those from wild-type mice. The exposure of cultured rat spermatogenic cells to ROS induced the release of cytochrome c from mitochondria, while Sertoli cells were more resistant under the same conditions. Tiron, a superoxide scavenger, suppressed the heat-induced release of cytochrome c from mitochondria. Collectively, these data suggest that ROS are generated during heat stress and cause spermatogenic cell death. Alternatively, since even a short exposure triggers harmful damage to spermatogenic cells, generated ROS may function as a type of signal for cell death rather than directly causing oxidative damage to cells. PMID- 16036349 TI - Antioxidant enzyme activity and lipid peroxidation in liver of female rats co exposed to lead and cadmium: effects of vitamin E and Mn2+. AB - The oxidative status of liver of female rats exposed to lead acetate and cadmium acetate either alone or in combination at a dose of 0.05 mg/kg body wt intraperitoneally for 15 days was studied. After the administration of lead alone, the activity of superoxide dismutase (SOD) decreased in liver, whereas no changes were observed in catalase (CAT) activity, and glutathione (GSH) and thiobarbituric acid (TBARS) levels. Cadmium exposure and combined exposure to lead and cadmium led to decrease in GSH content and increased TBARS levels. Moreover, animals exposed to either cadmium alone or in combination with lead showed a decrease in SOD activity and an increase in CAT activity. The in vitro experiments showed that vitamin E failed to restore the antioxidant enzyme activities in metal treated postmitochondrial supernatant fraction of liver. But Mn2+ ions protected the mitochondria from lipid peroxidation and could completely restore Mn-superoxide dismutase (Mn-SOD) activity following metal intoxication. The results of this study indicate that despite the ability of lead and cadmium to induce oxidative stress the effect in liver is not intensified by combined exposure to both lead and cadmium. The observed changes in various oxidative stress parameters in the liver of rats co-exposed to lead and cadmium may result from an independent effect of lead and cadmium and also from their interaction such as changes in metal accumulation and content of essential elements like Cu, Zn and Fe. These results suggest that when lead and cadmium are present together in similar concentrations, cadmium mediates major effects due to its more reactive nature. PMID- 16036350 TI - Effects of various vitamins and coenzymes Q on reactions involving alpha-hydroxyl containing radicals. AB - Effects of vitamins B, C, E, K and P, as well as coenzymes Q, on formation of final products of radiation-induced free-radical transformations of ethanol, ethylene glycol, alpha-methylglycoside and glucose in aqueous solutions were studied. Based on the obtained results, it can be concluded that there are substances among vitamins and coenzymes that effectively interact with alpha hydroxyl-containing radicals. In the presence of these substances, recombination reactions of alpha-hydroxyalkyl radicals and fragmentation of alpha-hydroxy-beta substituted organic radicals are suppressed. It has been established that the observed effects are due to the ability of the vitamins and coenzymes under study to either oxidize alpha-hydroxyl-containing radicals yielding the respective carbonyl compounds or reduce them into the initial molecules. PMID- 16036351 TI - Reactive nitrogen species formation in eosinophils and imbalance in nitric oxide metabolism are involved in atopic dermatitis-like skin lesions in NC/Nga mice. AB - Nitric oxide (NO) and reactive nitrogen species (RNS) have been implicated in the pathogenesis of inflammatory diseases. However, the involvement of NO and RNS in atopic dermatitis (AD), a pruritic inflammatory skin diseases, is not fully understood. In this study, we investigated the contribution of NO and RNS to the development of AD-like skin lesions in NC/Nga mice, an animal model for human AD. AD-like skin lesions were observed in NC/Nga mice kept under conventional conditions but not in specific pathogen-free conditions. The expression of inducible NO synthase (iNOS) and endothelial NOS (eNOS) proteins was upregulated in the dermal lesions, and that of neuronal NOS (nNOS) was downregulated in the epidermal lesions of the skin. Although the concentrations of NO2(-) and NO3(-) were lower, protein-bound nitrotyrosine content was significantly increased in the skin lesions. Immunohistochemical localization of nitrotyrosine was observed in almost all eosinophils. These results suggest that RNS formation in eosinophils and imbalance of NO metabolism are involved in the pathogenesis of AD like skin lesions in NC/Nga mice. PMID- 16036352 TI - Interaction of pyrogallol red with peroxyl radicals. A basis for a simple methodology for the evaluation of antioxidant capabilities. AB - A competitive method to evaluate the reactivity of highly reactive antioxidants is reported. Pyrogallol red (PGR) and AAPH (2,2'-azo-bis-(2-amidinopropane) dihydrochloride) were employed as target-molecule and peroxyl radical source, respectively. In the zero-order kinetic limit in PGR, the dependence of the ratio R(o)/R (where R(o) is the rate of the process in the absence of additive and R is the rate of the process in the presence of additive) upon the additive concentration (Stern-Volmer like plots) was studied. Various polyphenols (n=10) and ascorbic acid (AA) were tested as additives. In PGR protection by AA, was observed a neat induction time, associated to the total protection of the target molecule. On the other hand, the experiments that were carried out in presence of phenolic compounds allowed a relative evaluation of their reactivity towards peroxyl radicals. This reactivity follows the order quercetin > gallic acid > Trolox > kaempferol. Data obtained employing quercetin and Trolox are compatible with a competitive protection by these antioxidants. Due to the high reactivity of PGR towards peroxyl radicals and its high extinction coefficient at long wavelengths, it is a very suitable molecule to be employed as target in the evaluation of the free radical scavenging capability of very reactive phenolic compounds. PMID- 16036353 TI - Production of nitric oxide-derived reactive nitrogen species in human oral cavity and their scavenging by salivary redox components. AB - Nitrite is reduced to nitric oxide (NO) in the oral cavity. The NO generated can react with molecular oxygen producing reactive nitrogen species. In this study, reduction of nitrite to NO was observed in bacterial fractions of saliva and whole saliva. Formation of reactive nitrogen species from NO was detected by measuring the transformation of 4,5-diaminofluorescein (DAF-2) to triazolfluorescein (DAF-2T). The transformation was fast in bacterial fractions but slow in whole saliva. Salivary components such as ascorbate, glutathione, uric acid and thiocyanate inhibited the transformation of DAF-2 to DAF-2T in bacterial fractions without affecting nitrite-dependent NO production. The inhibition was deduced to be due to scavenging of reactive nitrogen species, which were formed from NO, by the above reagents. The transformation of DAF-2 to DAF-2T was faster in bacterial fractions and whole saliva which were prepared 1-4 h after tooth brushing than those prepared immediately after toothbrushing. Increase in the rate as a function of time after toothbrushing seemed to be due to the increase in population of bacteria which could reduce nitrite to NO. The results obtained in this study suggest that reactive nitrogen species derived from NO are continuously formed in the oral cavity and that the reactive nitrogen species are effectively scavenged by salivary redox components in saliva but the scavenging is not complete. PMID- 16036354 TI - Ischemic preconditioning decreases the reperfusion-related formation of hydroxyl radicals in a rabbit model of regional myocardial ischemia and reperfusion: the role of K(ATP) channels. AB - The objective of this study was to assess the effects of ischemic preconditioning (IP) on hydroxyl free radical production in an in vivo rabbit model of regional ischemia and reperfusion. Another goal was to determine whether K(ATP) channels are involved in these effects. The hearts of anesthetized and mechanically ventilated New Zealand White rabbits were exposed through a left thoracotomy. After i.v. salicylate (100 mg/kg) administration, all animals underwent a 30-min stabilization period followed by 40 min of regional ischemia and 2 h of reperfusion. In the IP group, IP was elicited by 5 min of ischemia followed by 10 min of reperfusion (prior to the 40-min ischemia period). Glibenclamide, a K(ATP) channel blocker, was administered prior to the preconditioning stimulus. Infarct size was measured by 2,3,5-triphenyl tetrazolium chloride (TTC) staining. We quantified the hydroxyl-mediated conversion of salicylate to its 2,3 and 2,5 dihydroxybenzoate derivatives during reperfusion by high performance liquid chromatography coupled with electro-chemical detection.IP was evidenced by reduced infarct size compared to control animals: 22% vs. 58%, respectively. Glibenclamide inhibited this cardioprotective effect and infarct size was 53%. IP limited the increase in 2,3 and 2,5-dihydroxybenzoic acid to 24.3 and 23.8% above baseline, respectively. Glibenclamide abrogated this effect and the increase in 2,3 and 2,5-dihydroxybenzoic acid was 94.3 and 85% above baseline levels, respectively, similar to the increase in the control group. We demonstrated that IP decreased the formation of hydroxyl radicals during reperfusion. The fact that glibenclamide inhibited this effect, indicates that K(ATP) channels play a key role in this cardioprotective effect of IP. PMID- 16036355 TI - Superoxide dismutase (SOD) as a potential inhibitory mediator of inflammation via neutrophil apoptosis. AB - Superoxide dismutase (SOD) is supposed to be an effective agent for neutrophil mediated inflammation in the area of critical medicine. We investigated the involvement of SOD in the regulation of neutrophil apoptosis. Exogenously added SOD effectively induced neutrophil apoptosis, and the fluorescence patterns determined using annexin-V and the 7-AAD were similar to those seen in Fas mediated neutrophil apoptosis. Neutrophils are short-lived leukocytes that need to be removed safely by apoptosis. The clearance of apoptotic neutrophils from sites of inflammation is a crucial determinant of the resolution of inflammation. Catalase inhibited the neutrophil apoptosis and caspase-3 activation. Spontaneous apoptosis, hydrogen peroxide and anti-Fas antibody-induced apoptosis of neutrophils were accelerated in Down's syndrome patients, in whom the SOD gene is overexpressed. Hydrogen peroxide was thought to be a possible major mediator of ROS-induced neutrophil apoptosis in caspase-dependent manner. Neutrophil apoptosis represents a crucial step in the mechanism governing the resolution of inflammation and has been suggested as a possible target for the control of neutrophil-mediated tissue injury. SOD may be a potential inhibitory mediator of neutrophil-mediated inflammation. PMID- 16036356 TI - Serum selenium predicts levels of F2-isoprostanes and prostaglandin F2alpha in a 27 year follow-up study of Swedish men. AB - Low concentrations of selenium (Se) predict mortality and cardiovascular diseases in some populations. The effect of Se on in vivo indicators of oxidative stress and inflammation, two important features of atherosclerosis, in human populations is largely unexplored. This study investigated the longitudinal association between serum selenium (s-Se) and a golden standard indicator of oxidative stress in vivo (8-iso-prostaglandin F2alpha, a major F2-isoprostane), an indicator of cyclooxygenase (COX)-mediated inflammation (prostaglandin F2alpha), high sensitive C-reactive protein (hsCRP), interleukin-6 (IL-6) and serum amyloid A protein (SAA) in a follow-up study of 27 years. The s-Se was measured in 615 Swedish men at 50 years of age in a health investigation. The status of oxidative stress and inflammation was evaluated in a re-investigation 27 years later by quantification of urinary 8-iso-PGF2alpha and 15-keto-dihydro-PGF2alpha (a major metabolite of PGF2alpha) and serum hsCRP, SAA and IL-6. Men in the highest quartile of s-Se at age 50 had decreased levels of 8-iso-PGF2alpha compared to all lower quartiles and decreased levels of PGF2alpha compared to all lower quartiles at follow-up. These associations were independent of BMI, diabetes, hyperlipidemia, hypertension, smoking, alpha-tocopherol and beta-carotene at baseline. The s-Se was not associated with hsCRP, SAA or IL-6 at follow-up. In conclusion, high concentrations of s-Se predict reduced levels of oxidative stress and subclinical COX-mediated (but not cytokine-mediated) inflammation in a male population. The associations between Se, oxidative stress and inflammation, respectively, might be related to the proposed cardiovascular protective property of Se. PMID- 16036357 TI - Oxidative stress in myotonic dystrophy type 1. AB - Myotonic dystrophy type 1 (DM1) is the most common form of muscular dystrophy affecting adults. The genetic basis of DM1 consists of a mutational expansion of a repetitive trinucleotide sequence (CTG). The number of triplets expansion divides patients in four categories related to the molecular changes (E1, E2, E3, E4). The pathogenic mechanisms of multi-systemic involvement of DM1 are still unclear. DM1 has been suspected to be due to premature aging, that is known to be sustained by increased free radicals levels and/or decreased antioxidants activities in neurodegenerative disorders. Recently, the gain-of-function at RNA level hypothesis has gained great attention, but oxidative stress might act in the disease progression. We have investigated 36 DM1 patients belonging to 22 unrelated families, 10 patients with other myotonic disorders (OMD) and 22 age matched healthy controls from the clinical, biochemical and molecular point of view. Biochemical analysis detected blood levels of superoxide dismutase (SOD), malonilaldehyde (MDA), vitamin E (Vit E), hydroxyl radicals (OH) and total antioxidant system (TAS). Results revealed that DM1 patients showed significantly higher levels of SOD (+40%; MAL (+57%; RAD 2 (+106%; and TAS (+20%; than normal controls. Our data support the hypothesis of a pathogenic role of oxidative stress in DM1 and therefore confirm the detrimental role played by free radicals in this pathology and suggest the opportunity to undertake clinical trials with antioxidants in this disorder. PMID- 16036358 TI - Evaluation of activity of selected antioxidants on proteins in solution and in emulsions. AB - Protection against protein oxidation by lipophilic and hydrophilic antioxidants in model systems using bovine serum albumin (BSA) in solution alone, or in an emulsion with linolenic acid methyl ester (LnMe) was found to be strongly dependent on the oxidation initiator. Tocopherol, Trolox, or the carotenoids astaxanthin and canthaxanthin were incubated with BSA or BSA/LnMe and oxidation was initiated either with the water-soluble azo-initiator 2,2' azo-bis-(2 amidinopropane) hydrochloride (AAPH), or FeCl3 and ascorbate, or the Fenton system using FeCl2/EDTA/H2O2, or with the singlet oxygen generating species anthracene-9,10-dipropionic acid disodium 1,4 endoperoxide (NDPO2). The results show that all the antioxidants tested were inefficient in the system with FeCl3/ascorbate. However, with the other initiating agents, the hydrophilic antioxidant, Trolox, was the most effective in preventing both protein and lipid oxidation. In contrast the lipophilic antioxidants were ineffective in preventing oxidation of BSA in aqueous solution, but did show some moderate antioxidative activity on protein and lipid in the BSA/LnMe system. Using the singlet oxygen generating system it was also demonstrated that Trolox always provided better protection of the protein than tocopherol and the carotenoids in both the BSA and the BSA/LnMe systems. In conclusion, prevention of protein oxidation using a water-soluble antioxidant has a protective effect on the lipid fraction and this approach deserves further attention in complex biological systems. PMID- 16036359 TI - DNA protecting and genotoxic effects of olive oil related components in cells exposed to hydrogen peroxide. AB - In search for compounds, able to protect nuclear DNA in cells exposed to oxidative stress, extracts from olive leaves, olive fruits, olive oil and olive mill waste water were tested by using the "single cell gel electrophoresis" methodology (comet assay). Jurkat cells in culture were exposed to continuously generated hydrogen peroxide (11.8+/-1.5 microM per min) by direct addition into the growth medium of the appropriate amount of the enzyme "glucose oxidase" in the presence or absence of the tested total extracts. The protective effects of the tested extracts or isolated compounds were evaluated from their ability to decrease hydrogen peroxide-induced formation of single strand breaks in the nuclear DNA, while the toxic effects were estimated from the increase of DNA damage when the extracts or isolated compounds were incubated directly with the cells. Significant protection was observed in extracts from olive oil and olive mill waste water. However, above a concentration of 100 microg/ml olive oil extracts exerted DNA damaging effects by themselves in the absence of any H2O2. Extracts from olive leaves and olive fruits although protective, were also able to induce DNA damage by themselves. Main compounds isolated from the above described total extracts, like oleuropein glucoside, tyrosol, hydroxytyrosol and caffeic acid, were tested in the same experimental system and found to exert cytotoxic (oleuropein glucoside), no effect (tyrosol) or protective effects (hydroxytyrosol and caffeic acid). In conclusion, cytoprotective as well as cytotoxic compounds with potential pharmaceutical properties were detected in extracts from olive oil related sources by using the comet assay methodology. PMID- 16036360 TI - Methods of quantitative analysis of the nitric oxide metabolites nitrite and nitrate in human biological fluids. AB - In human organism, the gaseous radical molecule nitric oxide (NO) is produced in various cells from L-arginine by the catalytic action of NO synthases (NOS). The metabolic fate of NO includes oxidation to nitrate by oxyhaemoglobin in red blood cells and autoxidation in haemoglobin-free media to nitrite. Nitrate and nitrite circulate in blood and are excreted in urine. The concentration of these NO metabolites in the circulation and in the urine can be used to measure NO synthesis in vivo under standardized low-nitrate diet. Circulating nitrite reflects constitutive endothelial NOS activity, whereas excretory nitrate indicates systemic NO production. Today, nitrite and nitrate can be measured in plasma, serum and urine of humans by various analytical methods based on different analytical principles, such as colorimetry, spectrophotometry, fluorescence, chemiluminescence, gas and liquid chromatography, electrophoresis and mass spectrometry. The aim of the present article is to give an overview of the most significant currently used quantitative methods of analysis of nitrite and nitrate in human biological fluids, namely plasma and urine. With minor exception, measurement of nitrite and nitrate by these methods requires method dependent chemical conversion of these anions. Therefore, the underlying mechanisms and principles of these methods are also discussed. Despite the chemical simplicity of nitrite and nitrate, accurate and interference-free quantification of nitrite and nitrate in biological fluids as indicators of NO synthesis may be difficult. Thus, problems associated with dietary and laboratory ubiquity of these anions and other preanalytical and analytical factors are addressed. Eventually, the important issue of quality control, the use of commercially available assay kits, and the value of the mass spectrometry methodology in this area are outlined. PMID- 16036361 TI - Extracellular H2O2 and not superoxide determines the compartment-specific activation of transferrin receptor by iron regulatory protein 1. AB - Iron regulatory protein 1 (IRP1) functions as translational regulator that plays a central role in coordinating the cellular iron metabolism by binding to the mRNA of target genes such as the transferrin receptor (TfR)--the major iron uptake protein. Reactive oxygen species such as H2O2 and O2*- that are both co released by inflammatory cells modulate IRP1 in opposing directions. While H2O2- similar to iron depletion--strongly induces IRP1 via a signalling cascade, O2*- inactivates the mRNA binding activity by a direct chemical attack. These findings have raised the question of whether compartmentalization may be an important mechanism for isolating these biological reactants when released from inflammatory cells during the oxygen burst cascade. To address this question, we studied cytosolic IRP1 and its downstream target TfR in conjunction with a tightly controlled biochemical modulation of extracellular O2*- and H2O2 levels mimicking the oxygen burst cascade of inflammatory cells. We here demonstrate that IRP1 activity and expression of TfR are solely dependent on H2O2 when co released O2*- with from xanthine oxidase. Our findings confirm that extracellular H2O2 determines the functionality of the IRP1 cluster and its downstream targets while the reactivity of O2*- is limited to its compartment of origin. PMID- 16036362 TI - Characterization of the high-resolution ESR spectra of superoxide radical adducts of 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline N-oxide (DEPMPO) and 5,5-dimethyl 1-pyrroline N-oxide (DMPO). Analysis of conformational exchange. AB - It has been previously reported that the spin trap 5-(diethoxyphosphoryl)-5 methyl-1-pyrroline N-oxide (DEPMPO) can form stable radical adducts with superoxide radical. However, the presence of diastereomers of DEPMPO radical adducts and the appearance of superhyperfine structure complicates the interpretation of the ESR spectra. It has been suggested that the superhyperfine structure in the ESR spectrum of DEPMPO/*OOH is a result of conformational exchange between conformers. The analysis of the temperature dependence of the ESR spectrum of DEPMPO/*OOH and of its structural analog DMPO/*OOH have demonstrated that both ESR spectra contain exchange effects resulting from conversion between two conformers. Computer simulation calculates a conformer lifetime on the order of 0.1 micros for DMPO/*OOH at room temperature. However, temperature dependence of the ESR spectrum of DEPMPO/*OOH suggests that superhyperfine structure does not depend on the conformational exchange. We have now found that the six-line ESR spectrum with superhyperfine structure should be assigned to a DEPMPO-superoxide-derived decomposition product. Therefore, ESR spectra previously assigned to DEPMPO/*OOH contain not only the two diastereomers of DEPMPO/*OOH but also the decomposition product, and these spectra should be simulated as a combination of four species: two conformers of the first diastereomer, one conformer of the second diastereomer and the superoxide-derived decomposition product. The presence of four species has been supported by the temperature dependence of the ESR spectra, nucleophilic synthesis of radical adducts, and isotopic substitution experiments. It is clear that to correctly interpret DEPMPO spin trapping of superoxide radicals, one must carefully consider formation of secondary radical adducts. PMID- 16036363 TI - Leukocyte 8-oxo-7,8-dihydro-2'-deoxyguanosine and comet assay in epirubicin treated patients. AB - Epirubicin fights cancer through topoisomerase II inhibition, hence producing DNA strand breaks that finally lead to cell apoptosis. But anthracyclines produce free radicals that may explain their adverse effects. Dexrazoxane--an iron chelator--was proven to decrease free radical production and anthracycline cardiotoxicity. In this article, we report the concentrations of cellular 8-oxo 7,8-dihydro-2'-deoxyguanosine (8-oxo-dGuo) relative to 2'-deoxyguanosine (dGuo), and comet assay results from a study including 20 cancer patients treated with epirubicin. Plasma concentrations of vitamins A, E, C and carotenoids are also reported. All data were obtained before and immediately after epirubicin infusion. The ratios of 8-Oxo-dGuo to dGuo were measured in leukocyte DNA by HPLC coulometry after NaI extraction of nucleic acids. Vitamins A and E and carotenoids were measured by HPLC-spectrophotometry. Vitamin C was measured by HPLC-spectrofluorimetry. Median 8-oxo-dGuo/dGuo ratios increased significantly from 0.34 to 0.48 lesions per 100,000 bases while per cent of tail DNA increased from 3.47 to 3.94 after chemotherapy 8-Oxo-dGuo/dGuo and per cent of tail DNA medians remained in the normal range. Only vitamin C decreased significantly from 55.4 to 50.3 microM Decreases in vitamins A, E, lutein and zeaxanthin were not significant, but concentrations were below the lower limit of the normal range both before and after chemotherapy. Only the correlation between comet assay results and vitamin C concentrations was significant (rho =-0.517, p = 0.023). This study shows that cellular DNA is damaged by epirubicin-generated free radicals which produce the mutagenic modified base 8-oxo-dGuo and are responsible for strand breaks. However, strand breaks are created not only by free radicals but also by topoisomerase II inhibition. In a previous study we did not find any significant change in urinary 8-oxo-dGuo excretion after adriamycin treatment. However, 8-oxo-dGuo may have increased at the end of urine collection as DNA repair and subsequent kidney elimination are relatively slow processes. In another study, authors used GC-MS to detect 8-oxo-dGuo in DNA and did not find any change after prolonged adriamycin infusion. Reasons for these apparent discrepancies are discussed. PMID- 16036364 TI - Rapid increase in serum lipid peroxide 4-hydroxynonenal (HNE) through monocyte NADPH oxidase in early endo-toxemia. AB - We have developed a time-resolved fluoroimmunoassay (TR-FIA) for a lipid peroxide 4-hydroxynonenal (HNE), which is 100-fold more sensitive than conventional enzyme linked immunosorbent assay (ELISA) and is an easier technique to use for a large number of samples without pre-treatment. By this assay, we found that a low dose of bacterial lipo-polysaccharide (LPS), injected intra-peritoneally (0.5 mg/kg), increased serum HNE level by 28-folds, with a peak at 20 min. LPS also increased HNE in vitro to a much higher level in the monocyte-enriched plasma than in the leukocyte-enriched plasma, with a peak at 10 min. The HNE production after LPS treatment was inhibited by apocynin, a specific NADPH oxidase inhibitor in vivo and in vitro, and to a lesser extent by dimethylsulfoxide a solvent for apocynin and a hydroxyl radical scavenger in vitro. These data suggest that monocyte NADPH oxidase is involved in the lipid peroxidation (HNE formation) in the LPS challenged rat. This is the first clear demonstration of the link between an inflammatory stimulus and lipid peroxidation in the blood. PMID- 16036365 TI - Superoxide and hydrogen peroxide formation during enzymatic oxidation of DOPA by phenoloxidase. AB - Generation of superoxide anion and hydrogen peroxide during enzymatic oxidation of 3-(3,4-dihydroxyphenyl)-DL-alanine (DOPA) has been studied. The ability of DOPA to react with O2*- has been revealed. EPR spectrum of DOPA-semiquinone formed upon oxidation of DOPA by O2*- was observed using spin stabilization technique of ortho-semiquinones by Zn2+ ions. Simultaneously, the oxidation of DOPA by O2*- was found to produce hydrogen peroxide (H2O2). The analysis of H2O2 formation upon oxidation of DOPA by O2*- using 1-hydroxy-3-carboxy-pyrrolidine (CP-H), and SOD as competitive reagents for superoxide provides consistent values of the rate constant for the reaction between DOPA and O2*- being equal to (3.4+/ 0.6)x10(5) M(-1) s(-1). The formation of H2O2 during enzymatic oxidation of DOPA by phenoloxidase (PO) has been shown. The H2O2 production was found to be SOD sensitive. The inhibition of H2O2 production by SOD was about 25% indicating that H2O2 is produced both from superoxide anion and via two-electron reduction of oxygen at the enzyme. The attempts to detect superoxide production during enzymatic oxidation of DOPA using a number of spin traps failed apparently due to high value of the rate constant for DOPA interaction with O2*-. PMID- 16036366 TI - Plasma cytokines and oxidative damage in HIV-positive and HIV-negative adolescents and young adults: a protective role for IL-10? AB - HIV infection causes immune activation that leads to oxidative damage. Proinflammatory cytokines may promote such damage and the regulatory cytokine IL 10 may protect against such damage. To examine the relation of these cytokines to oxidative damage, 67 cases of oxidative damage and 67 matched controls were selected from the reaching for excellence in adolescent health (REACH) study. Subjects were young (15-23 years), largely female (76%), HIV-positive (73%) and black (69%). Proinflammatory cytokines were not significantly associated with oxidative damage but plasma IL-10 had a significant, negative association with oxidative damage. This finding is consistent with a protective role for IL-10 in diminishing oxidative damage during immune activation. PMID- 16036367 TI - Effect of arachidonic, eicosapentaenoic and docosahexaenoic acids on the oxidative status of C6 glioma cells. AB - n-3 polyunsaturated fatty acids (PUFAs) have been described to have beneficial effects on brain development and in the prevention and treatment of brain damage. C6 glioma cells were incubated with 100 microM of either C20:4n-6 (ARA), or C20:5n-3 (EPA), or C22:6n-3 (DHA) for different time periods to assess whether these acids altered the cellular oxidative state. The ARA and EPA were promptly metabolised to C22:4n-6 and C22:5n-3, respectively, whereas DHA treatment simply increased the amount of DHA in the cells. Cell viability was not affected by ARA, while a cytotoxic effect was observed 72 h after n-3 PUFAs supplementation. The levels of reactive oxygen species and thiobarbituric acid-reactive substances were significantly higher in DHA-treated cells than in EPA- and ARA-treated groups. This modification in the oxidative cellular status was also highlighted by a significant increase in catalase activity and a decrease in glutathione content in DHA-supplemented cells. Glucose-6-phosphate dehydrogenase activity, an enzyme involved in redox regulation, and O2*- release were significantly increased both in EPA and DHA groups. The effect of DHA was more severe than that of EPA. No significant changes were observed in the ARA group with respect to untreated cells. These data show that EPA and DHA induce alterations in the oxidative status that could affect the glial function. PMID- 16036368 TI - Iron release analyses from ferritin by visible light irradiation. AB - We investigated the iron release from ferritin by irradiation from a white fluorescent light in the absence or presence of ADP. Irradiation of a ferritin solution at 17,000 lx in the absence of ADP slightly induces iron release from ferritin but only at acidic pH conditions (pH 5.0 or pH 6.0). Irradiation in the presence of ADP markedly enhances iron release from ferritin under the same conditions. In the absence of irradiation, the iron release from ferritin was low even in the presence of ADP. The induction of the iron release by irradiation in the presence of ADP was also affected by various factors such as irradiation dose and acidity, but not temperature (4-47 degrees C), oxygen concentration, or free radical generations during the irradiation. The iron release during the irradiation ceased to increase by turning off the light and was found to increase again after additional irradiation. These results suggest that visible light directly induces iron release from ferritin via the photoreduction of iron stored inside ferritin. PMID- 16036369 TI - Triphlorethol-A from Ecklonia cava protects V79-4 lung fibroblast against hydrogen peroxide induced cell damage. AB - In the present study, triphlorethol-A, a phlorotannin, was isolated from Ecklonia cava and its antioxidant properties were investigated. Triphlorethol-A was found to scavenge intracellular reactive oxygen species (ROS) and 1,1-diphenyl-2 picrylhydrazyl (DPPH) radical, and thus prevented lipid peroxidation. The radical scavenging activity of triphlorethol-A protected the Chinese hamster lung fibroblast (V79-4) cells exposed to hydrogen peroxide (H2O2) against cell death, via the activation of ERK protein. Furthermore, triphlorethol-A reduced the apoptotic cells formation induced by H2O2. Triphlorethol-A increased the activities of cellular antioxidant enzymes like, superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Hence, from the present study, it is suggestive that triphlorethol-A protects V79-4 cells against H2O2 damage by enhancing the cellular antioxidative activity. PMID- 16036370 TI - Effect of vegetables on human phenolsulfotransferases in relation to their antioxidant activity and total phenolics. AB - Epidemiology studies have shown that consumption of fruits and vegetables is associated with the prevention of chronic diseases such as cancer and cardiovascular disease. Induction of cellular phase II detoxifying enzymes is associated with cancer preventive potential. Phenolsulfotransferases (PSTs) are traditionally known as phase II drug-metabolizing or detoxifying enzymes that facilitate the removal of drugs and other xenobiotic compounds. Phenolic acids are known to increase the activities of PSTs. In the present study, human HepG2 cells were used as model to investigate the influence of twenty vegetables on human PST activity and to evaluate the relationships to their antioxidant activity and total phenolics content. The result showed that PST-P activity was significantly (p < 0.01) induced by asparagus, broccoli, cauliflower, celery and eggplant, whereas PST-M activity was induced by asparagus, broccoli, carrot, eggplant and potato at a concentration of 100 microg/ml. The vegetable extracts that induced both forms of PSTs activities were found to have higher antioxidant capacities and total phenolic content in the oxygen radical absorbance capacity (ORAC) and Folin-Ciocalteu assay. The major polyphenols in broccoli, the most potential inducer in both forms of PSTs activities, was antioxidant phenolic acids. HPLC retention times and standard spiked indicated the presence of gallic acid, p-hydroxybenzoic acid, p-coumaric acid, gentisic acid and ferulic acid in broccoli. The overall effect of vegetables tested on the activity of PST-P was well correlated to their ORAC value and total phenolics content (r= 0.82, p < 0.05 and r = 0.78, p < 0.05). These results imply that vegetables have a capability of inducing PST activity, and the PST induction may be possibly ascribed to antioxidant phenolic acids in vegetable extracts. PMID- 16036372 TI - Regulatory T cells in autoimmune diseases: anti-ergotypic T cells. AB - T regulatory cells play an important role in regulating T-cell responses to self antigens and control autoimmunity and autoimmune disease. Anti-ergotypic T cells are a subset of such regulatory T cells that respond to activation markers, ergotopes, expressed on other activated T cells. Anti-ergotypic T cells do not respond to nonactivated T cells. Ergotopes include the a-chain of the IL-2 receptor (CD25). Anti-ergotypic T cells were found to downregulate experimental diseases such as experimental autoimmune encephalomyelitis (EAE) and adjuvant arthritis (AA). Anti-ergotypic T cells are present in humans and are activated after T-cell vaccination. Here we review anti-ergotypic T cells in animal models and in humans and contrast anti-ergotypic T cells with other regulatory T-cell subsets. PMID- 16036373 TI - Stress proteins as inducers and targets of regulatory T cells in arthritis. AB - Immunization with microbial or mammalian stress proteins or heat-shock proteins in models of experimental autoimmunity has been observed to lead to increased disease resistance. Furthermore, such immunization has been proposed to result in the induction and expansion of T cells that suppress disease upon transfer. Comparisons of microbial heat-shock proteins with other conserved immunogenic proteins of bacterial origin have indicated a unique capacity for heat-shock proteins to induce a regulatory phenotype in T cells, such as reflected by the production of IL10. Also, studies in children with chronic arthritis have indicated that T-cell responses to heat-shock proteins are associated with a benign course of the disease and with remission. Furthermore, in patients, heat shock-protein-(HSP-) activated T cells were shown to display regulatory phenotypes consistent with CD4+ CD25+ T regulatory cells. PMID- 16036374 TI - Feedback regulation of autoimmunity via TCR-centered regulation. AB - The complexity of a self-regulatory system demands a balance between effectors and regulators; that is, it is necessary for both cell types to exist. Regulation of self-reactive T cells can occur at several complementary but different levels: (1) at the level of priming itself: for example, inhibition of expansion of antigen-reactive T cells by regulatory CD4+ CD25+ T cells; (2) after the priming of self-reactive T cells, regulatory T-cell populations with reactivity to distinct self-determinants derived from the T-cell receptor (TCR) can be engaged via a negative feedback mechanism. Thus, these mechanisms ensure induction of effective and appropriately limited responses against foreign antigens while preventing autoreactivity from inflicting self-damage. PMID- 16036375 TI - Regulatory T cells in immunologic self-tolerance and autoimmune disease. AB - Naturally arising CD25+ CD4+ regulatory T cells play key roles in the maintenance of immunologic self-tolerance and negative control of various immune responses. The majority, if not all, of them are produced by the normal thymus as a functionally distinct T-cell subpopulation, and their generation is in part developmentally controlled. Genetic abnormality in the development and function of this population can indeed be a cause of autoimmune disease, immunopathology, and allergy in humans. This regulatory population can be exploited to prevent and treat autoimmune disease by strengthening and reestablishing immunologic self tolerance. PMID- 16036376 TI - Neonatal autoimmune disease: influence of CD4+ CD25+ regulatory T cells. AB - Although previous studies have emphasized the tolerogenic property of murine neonatal immune system, recent studies indicate that neonatal mice are prone to autoimmune disease. This chapter will summarize the evidence for neonatal propensity to autoimmune ovarian disease (AOD) and describe the new finding that autoantibody can trigger a T cell-dependent autoimmune disease in neonatal but not adult mice. Based on depletion or addition of the CD4+ CD25+ T cells, disease resistance of older mice is explicable by the emergence of CD4+ CD25+ regulatory T-cell function after day 5, whereas disease susceptibility is associated with resistance to regulation by CD4+ CD25+ T cells. PMID- 16036377 TI - Regulatory T cells in the control of autoimmune diabetes: the case of the NOD mouse. AB - Over the last few years, there has been a revival of the concept of suppressor/regulatory T cells being central players in the control of various immune responses, including autoimmune responses and immune response to transplants, tumors, and infectious agents. It appears that regulatory T cells are diverse in their phenotypes, antigen specificity, and modes of action. Here we summarize studies from various groups, including our own, demonstrating that specialized subsets of regulatory T cells are pivotal in the control of autoimmune diabetes as well shown by the compelling evidence accumulated using the non-obese diabetic (NOD) mouse model. We also provide a discussion of the evidence showing that some biological products (such as CD3-specific monoclonal antibodies) are representatives of a new category of immunotherapeutic agents endowed with unique capacities to promote immunological tolerance (an antigen specific unresponsiveness in the absence of long-term generalized immunosuppression) through their ability to induce immunoregulatory T cells. PMID- 16036378 TI - Do regulatory T cells play a role in the control of homeostatic proliferation? AB - The control of peripheral lymphocyte numbers is a fundamental aspect of the immune system. Regulatory T cells are involved in the suppression of autoimmune, antitumor, allergic, and other inflammatory responses, as well as in facilitating graft acceptance. In this paper, we discuss whether the control of homeostatic proliferation is another facet of the immune system that is controlled by regulatory T cells. A review of the published data connecting regulatory T cells with the control of homeostatic proliferation indicates that several key questions remain open. One of these relates to the stage at which regulatory T cells could play a role (i.e., T-cell proliferation vs. survival). PMID- 16036386 TI - The renin-aldosterone response to stimulation and suppression during normal pregnancy. AB - OBJECTIVE: During normal pregnancy, studies have shown increased activity of the renin-angiotensin-aldosterone system (RAAS) and a dissociation of plasma renin activity (PRA) and aldosterone (Aldo) evidenced by a greater increase in Aldo relative to PRA. The aims of this study were to examine the RAAS response to stimulation by upright posture and suppression by saline infusion and to investigate the PRA-Aldo dissociation under these two conditions. METHODS: We studied 24 healthy normotensive women (mean+/-standard error of mean, ages 29+/-1 yrs) in sodium (Na) balance in the second and third trimesters and postpartum. Subjects underwent a 24-hour urine collection which was analyzed for Na, norepinephrine (NE), epinephrine (Epi), and dopamine (DA); a posture study with analysis of blood pressure (BP), PRA, Aldo, NE, Epi, DA, and cortisol; and a 0.9% NaCl infusion study (500 mL/hr for 3 hrs) with analysis of BP, PRA, Aldo, cortisol, and digitalis-like factor (DLF). Analyses included paired t tests to compare posture and saline responses, repeated measures to compare across periods, and percent change to evaluate the PRA-Aldo dissociation. RESULTS: During pregnancy, PRA, Aldo, BP, catecholamines, and cortisol levels were significantly greater in upright than left lateral decubitus (LLD) posture, and the percent change in Aldo was significantly greater than the percent change in PRA. During pregnancy in response to saline infusion, BP did not change; the PRA and Aldo significantly decreased; the percent change in Aldo was significantly greater than the percent change in PRA in the second trimester; and serum DLF and cortisol levels significantly decreased. CONCLUSIONS: In longitudinally studied normal pregnancy, PRA and Aldo levels were dissociated at baseline, with stimulation and, to a lesser degree, with suppression. Norepinephrine, adrenocorticotrophic hormone, and DLF may contribute to this dissociation, and clarification of these interactions may provide insight into the regulation of aldosterone during normal and hypertensive pregnancy. PMID- 16036387 TI - Effect of magnesium on red blood cell deformability in pregnancy. AB - OBJECTIVE: Red blood cell (RBC) deformability is an important factor in determining movement of red blood cells through the microcirculation. In preeclampsia and some cases of intrauterine growth restriction (IUGR), RBC deformability and microcirculation are reduced. Magnesium is administered to reduce the risk of seizures. The aim of this study was first to detect the effect of intravenous magnesium application (2 g/h) on the deformability of RBCs in pregnancies with normal RBC deformability, receiving magnesium as tocolytic agent. The second aim was to examine the effect of calcium-antagonists (magnesium, nifedipin) on the deformability of RBC of preeclamptic patients in vitro. METHODS: Part 1: magnesium (2 g/h), fenoterol (270 microg/h)+verapamil (0.2 mg/h) or placebo (NaCl 0.9%) was administered intravenously to pregnant women with premature contractions to test the tocolytic effect. RBC-deformability was measured by laser diffractoscopy in all three groups. Blood samples were taken before, after 1 h and after 24 h of administration. Magnesium-plasma-levels were measured. Part 2: Blood samples from patients with preeclampsia were incubated in vitro with magnesium (2 mmol), nifedipine (0.25 mg/ml), or placebo (NaCl 0.9%). RBC deformability was measured before and 15 min, 1 h, 2 h, 6 h, and 10 h after start of the incubation. RESULTS: Part 1: The initial RBC deformability was the same in all groups (E=0.232+/-0.017 in NaCl, 0.232+/-0.023 in fenoterol+verapamil, 0.232+/-0.019 in magnesium). After 1 h of administration, RBC-deformability was significantly greater with magnesium (0.254+/-0.020) and Fenoterol+Verapamil (0.238+/-0.02) compared to placebo (0.231+/-0.015). After 24 h the effect on RBC deformability in the fenoterol+verapamil-group was gone (0.234+/-0.021 compared to 0.234+/-0.016 in placebo), while in the IV-magnesium group RBC-deformability remained increased (E=0.241+/-0.019). Statistical analysis of the influence of magnesium-plasma-levels showed the maximum effect at concentrations of 1.95-2.15 mmol/l. Part 2: RBC-deformability in preeclampsia was reduced as predicted by previous studies (0.120+0.0086 versus 0.232 in normal pregnancy). In vitro incubation with magnesium enhanced RBC-deformability in preeclampsia. Even after 15 min, a statistically significant effect was seen (0.127+/-0.0091 versus 0.121+/-0.0091 in placebo). Maximum effect was reached after 6 h of incubation (0.159+/-0.0093 versus 0.133+/-0.0091). Incubation with Nifedipine also enhanced RBC deformability [0.127+/-0.0091 after 15 min, 0.149+/ 0.010 after 6 h (maximum effect)], but the effect was less pronounced than with magnesium. CONCLUSION: Intravenous magnesium therapy over a 24-hour period increases RBC-deformability even in pregnancies with normal RBC-deformability. In vitro measurements show an increase of RBC-deformability in preeclampsia in response to magnesium, which could offer additional therapeutic benefit for the treatment of reduced blood flow seen in most cases of preeclampsia. PMID- 16036388 TI - Morphologic changes of the uteroplacental unit in preeclampsia-like syndrome in rats. AB - OBJECTIVE: Chronic administration of the nitric oxide synthase inhibitor NG-nitro L-arginine methylester (L-NAME) induces a preeclampsia-like syndrome in rats. This animal study aims to determine whether L-NAME-induced preeclampsia-like syndrome is also associated with morphologic changes in the uteroplacental unit. METHODS: Mating was induced in 20 adult Sprague-Dawley rats. On day 14, five pregnant rats were randomly assigned to receive L-NAME, whereas six served as controls. Weight, continuous blood pressure, urine volume, urine protein content, litter size, number of resorption sites, placental weight, and weight of pups were documented. Uteroplacental tissue of three L-NAME and three control animals were processed for microscopy using corrosion cast technique. RESULTS: The L-NAME treated animals had a significantly smaller litter size (p=.033) and a significantly higher number of resorption sites (p=.021) when compared to controls. In L-NAME-treated rats midlength inner-diameter of maternal arterial channels (p<.001) and cross-sectional area (p<.001) were significantly smaller than were those in controls. CONCLUSIONS: Chronic administration of L-NAME to pregnant rats is associated with significant morphologic changes of the uteroplacental vasculature. The L-NAME-induced pathologic alterations resulted in decreased litter size and increased number of resorption sites that may be contributed to diminished uteroplacental perfusion. PMID- 16036389 TI - Association study between the human Renin gene and preeclampsia. AB - OBJECTIVE: We recently reported on a missense mutation in exon 9 of the human renin gene (G1051A) that may affect the functioning of this enzyme, and is associated with essential hypertension. The aim of the present study is to assess the association between the genotypes of this missense mutation in the renin gene and preeclampsia (PE) via a case-control study. METHODS: DNA was extracted from peripheral blood leukocytes, and genotyping of G1051A was performed in 117 PE patients and in 171 non-PE controls. RESULTS: The frequency of genotypes for G1051A was not significantly different between the two groups. The frequency of the A1051 allele was also not significantly different between PE patients (52.6%) and non-PE controls (50.6%). CONCLUSIONS: The missense mutation G1051A in the human renin gene is not associated with PE. PMID- 16036390 TI - Maternal plasma homocysteine concentrations are not increased in twin pregnancies. AB - OBJECTIVE(S): We tested the hypothesis that twin pregnancies would lead to increased maternal plasma homocysteine. We further hypothesized that twin pregnancies complicated by preeclampsia would have increased plasma homocysteine compared to twin pregnancies without preeclampsia and normal singleton pregnancies. METHODS: Plasma was collected at delivery from 127 nulliparous subjects: 57 women with normal singleton pregnancies, 39 women with singleton and preeclampsia, 17 women with uncomplicated twin pregnancies, and 14 women with twins and preeclampsia. Subjects were group matched for prepregnancy body mass index (BMI) and race. Plasma homocysteine was analyzed by high pressure liquid chromatography (HPLC) with fluorescence detection, and plasma folic acid was measured by radio immunoassay (RIA). RESULTS: The mean plasma concentration of homocysteine was significantly increased in all women with preeclampsia (7.4+/ 2.9 microM) compared to all normal pregnant women (5.9+/-2.1 microM, p=0.002). However, homocysteine was not significantly increased in all women with twins (6.7+/-2.1 microM) compared to all women with singleton pregnancies (6.5+/-2.7 microM, p=0.61). In addition, women with twins and preeclampsia did not have increased homocysteine (6.8+/-2.1 microM) compared to women with twins and normal pregnancy (6.7+/-2.1 microM, p=0.72). As expected, because of extra supplementation, plasma folic acid was significantly increased in women with twins (27.9+/-11.6 ng/mL) compared to women with singleton pregnancies (20.8+/ 8.5 ng/mL, p=0.0003). However, folic acid was not different between preeclamptics and controls (23.5+/-10.8 vs. 21.9+/-9.2 ng/mL respectively, p=0.36). Lastly, there was a significant inverse correlation between homocysteine and folic acid among all the subjects (r2=- 0.053, p< 0.01), and this correlation persisted in the women with singleton pregnancies (r2=- 0.078, p< 0.01), but was lost in the twins (r2=- 0.073, p=0.14). CONCLUSIONS: With contemporary management including increased folic acid supplementation, plasma homocysteine is not increased in twin pregnancies with or without preeclampsia. PMID- 16036391 TI - Is high cerebral perfusion pressure and cerebral flow predictive of impending seizures in preeclampsia? A case report. AB - Transcranial Doppler ultrasound was used to demonstrate elevated estimated cerebral perfusion pressure (CPP) and cerebral flow index (CFI) in a preeclamptic patient. She subsequently developed eclampsia. After magnesium sulfate therapy her CPP and CFI were within the normal range and she did not experience further seizures. This finding suggests that cerebral overperfusion may be at least one of the etiologies involved in the pathogenesis of eclampsia. PMID- 16036392 TI - Placental system A transporter mRNA is not different in preeclampsia, normal pregnancy, or pregnancies with small-for-gestational-age infants. AB - OBJECTIVE: System A amino acid transporter activity is reduced in placentas from small-for-gestational-age (SGA) compared to normal pregnancies. We compared the expression of the system A transporters between preeclamptic and control and between small-for-gestational-age and controls pregnancies. METHODS: We used placental samples from 18 preeclamptic pregnancies matched with 17 normal pregnancies and from 16 SGA pregnancies matched with 15 different normal pregnancies. Using real-time reverse transcriptase-polymerase chain reaction (RT PCR) we quantified the mRNA for two system A subtype target genes ATA1 and ATA2 as well as beta-actin for normalization. RESULTS: There was no significant difference of mRNA for ATA1 or ATA2 transporters between preeclamptic and their controls or SGA pregnancies and their controls. CONCLUSIONS: Despite previous studies reporting reduced activity for system A transporters in small-for gestational-age pregnancies, we found no difference in steady-state concentrations of the mRNA, of the system A transporters among preeclamptic, SGA, and normal control pregnancies. These results do not exclude differences in actual protein levels or activity of the amino acid transporters, which warrant further study. PMID- 16036393 TI - Villous explant culture: characterization and evaluation of a model to study trophoblast invasion. AB - The mechanisms that control invasion of cytotrophoblast (CTB) cells into the maternal decidua and myometrium with transformation of the maternal spiral arteries are not fully understood, but oxygen is thought to be a key factor. We carried out a semiquantitative evaluation of an explant culture model for use in the study of trophoblast proliferation and invasion. Explants of human villous tissue (6-9 weeks of gestation) cultured on Matrigel in both standard culture conditions (18% O2) and in a low oxygen environment (2% O2) produced regions of outgrowth, of cytotrophoblast cells from villous tips and migration of cells into the Matrigel. The number of sites of outgrowth and migration, area of outgrowth, and extent of migration of cells into the Matrigel tended to increase throughout the culture period (144 h) but varied between explants from the same placenta and those from different placentas. There were no significant differences in the number of sites of outgrowth or migration scores in explants cultured in a low oxygen environment compared to those cultured in standard conditions. This study highlights the importance of careful validation, design and interpretation of experiments using in vitro culture systems, particularly those investigating the regulatory role of oxygen. PMID- 16036394 TI - Immediate blood pressure changes and aquatic physiotherapy. AB - OBJECTIVE: No studies to date have examined the effect on Blood pressure (BP) when pregnant women participate in aquatic physiotherapy (AP) classes; although, previous studies have indicated that there may be a detrimental outcome to the pregnant mother and fetus when there is a significant rise in the maternal BP. At least three components may contribute to BP changes: temperature, exercise, and immersion. This study examines BP changes in pregnant women attending AP classes. METHOD: An experimental same-subject, repeated measures design was employed. Forty-five sets of data were collected from pregnant women attending routine, existing, nonaerobic AP classes. Blood pressure measurements were taken preimmersion, 2-5 minutes after entering the pool, immediately following the 50 minute AP class (while immersed), and 8-12 minutes after exiting the water. RESULTS: Systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean arterial pressure (MAP) data were analyzed using a repeated measures analysis of variance (ANOVA). For all sets of data a highly significant difference was observed (p = 0.0000). Post-hoc Tukey test results showed BP to decrease significantly on entering the water (MAP mean difference of 10.52 mm Hg, 95% CI 8.75 to 12.29), to remain at this low level postexercise while still immersed, and then return to preimmersion values 8-12 minutes after exiting the water. CONCLUSION: These results indicate that AP classes are safe for nonhypertensive pregnant women in their third trimester. The use of AP in the antenatal care of women with unstable BP or who are hypertensive needs further research. PMID- 16036395 TI - MMP-2 levels are elevated in the plasma of women who subsequently develop preeclampsia. AB - OBJECTIVE: To determine levels of matrix metalloproteinase (MMP)-2 and MMP-9, and the tissue inhibitors of metalloproteinases (TIMP)-1 and TIMP-2 in the plasma of women destined to develop preeclampsia prior to the onset of clinical disease. STUDY DESIGN: Plasma samples were taken from women whose pregnancies were subsequently complicated by preeclampsia and from normal pregnant women at 22 and 26 weeks and at delivery or diagnosis. Following equal protein loading, MMP-2 and 9 and TIMP-1 and 2 were quantified using zymography and Western blot analysis, respectively. RESULTS: Plasma MMP-2 levels were significantly elevated at 22 weeks (p = 0.02) and at diagnosis (p = 0.003) in the preeclampsia group, but there was no difference at 26 weeks. TIMP-1 levels were significantly reduced in the preeclampsia group at 26 weeks (p = 0.0002), but TIMP-2 levels were not quantifiable. CONCLUSION: At all three gestational time points an imbalance in the MMP-2:TIMP-1 ratio was found in patients who subsequently developed preeclampsia. We speculate that increased net MMP-2 activity may contribute to the endothelial dysfunction that is central to the pathophysiology of preeclampsia. PMID- 16036396 TI - The accuracy of urine dipsticks as a screening test for proteinuria in hypertensive disorders of pregnancy. AB - BACKGROUND: Proteinuria is used as a criterion in the classification system for hypertensive disorders of pregnancy including preeclampsia. The aim of the study was to evaluate the accuracy of dipstick urinalysis in a single voided urine sample and in an aliquot of a 24-hour urine collection in the assessment of proteinuria in hypertensive pregnant women, using the 24-hour urine protein excretion as the gold standard. METHODS: One hundred ninety-eight women who presented with hypertension in pregnancy were recruited at the antenatal clinic at King Edward VIII Hospital in Durban, South Africa, a tertiary referral center. Exclusion criteria included women with eclampsia, urinary tract infection, and chronic renal disease. Routine dipstick urinalysis (Bayer) was performed by midwives for proteinuria, and a 24-hour urine specimen was collected for quantitative protein assessment. A laboratory technician performed urine dipstick test for protein on a mixed aliquot of the 24-hour urine specimen. This result, together with that of the screening dipstick urinalysis, was compared to the 24 hour urine protein excretion. RESULTS: The results of the 198 patients were analyzed, of the total, 72 had preeclampsia. Using a value of = 0.3 g protein excretion per 24 hours (1 + to 4 + on urine dipsticks) as positive, sensitivity, specificity, and predictive values for dipstick urinalysis were calculated. The positive predictive value for dipstick urinalysis ranged from 64.9% (single voided urine sample) to 94.2% (24-hour urine aliquot). The negative predictive value ranged from 75.2% (single voided urine sample) to 84.2% (24-hour urine aliquot). CONCLUSION: Dipstick urinalysis is not very accurate: therefore, all women presenting with hypertension during pregnancy should have a 24-hour urine protein measurement. PMID- 16036397 TI - The association between maternal obesity, glucose intolerance and hypertensive disorders of pregnancy in nondiabetic pregnant women. AB - OBJECTIVE: The aim of this study is to evaluate whether pregnancy-induced hypertension (PIH) among nondiabetic patients is associated with glucose intolerance. MATERIALS AND METHODS: A retrospective case-control study was designed including a study group who had pregnancy-induced hypertension or preeclampsia. Patients with normal pregnancy were used as a control group matched to cases by parity. Diabetic patients, nonsingleton pregnancies, and women without prenatal care were excluded. Data concerning fasting glucose levels, glucose challenge test (GCT), and oral glucose tolerance test (OGTT) were collected from patients' files. RESULTS: There were 131 patients in each study group. The study group had significantly higher mean maternal age, mean GCT levels, and mean pregestational body mass index (BMI) (28.0 +/- 5.8 vs. 26.5 +/- 5.3, p = 0.02; 5.8 +/- 1.4 vs. 5.1 +/- 1.1 p = 0.0018; 26 +/- 5.1 vs. 23 +/- 4.0 p < 0.001, respectively) than the control group. Mean gestational age and birthweight were also significantly lower in the study group (38.5 +/- 2.1 vs. 39.4 +/- 1.7 p < 0.001; 2929 g +/- 614.7 vs. 3225 +/- 461.1 p < 0.001, respectively). Stratified analysis according to parity demonstrated that pregestational BMI, weight gain during pregnancy, and cesarean section (CS) were significantly higher in women with pregnancy-induced hypertension than in controls in all parity groups. Maternal age and mean GCT levels of women with pregnancy-induced hypertension were higher in all parity groups but statistically significant only among multiparous patients. Multiple logistic regression demonstrated that BMI, weight gain, and maternal age were independently associated with pregnancy-induced hypertension, while GCT level was not. Conclusions. Elevated pregestational BMI is an independent risk factor for development of pregnancy-induced hypertension (PIH). Its association with elevated GCT levels implies that even without overt diabetes, glucose intolerance may play a role in the pathogenesis of preeclampsia in obese patients. PMID- 16036398 TI - A prospective analysis of the role of uterine artery Doppler waveform notching in the assessment of at-risk pregnancies. AB - OBJECTIVE: To determine the value of second trimester uterine artery Doppler waveform notching in the prediction of adverse pregnancy outcome in a high-risk group. DESIGN: Analysis of data from a consecutively collected cohort. SETTING: St. James University Hospital, Leeds, UK. POPULATION: Three hundred thirty women known to be at risk of preeclampsia (PET) or intrauterine growth restriction (IUGR) were assessed for notching of the uterine artery Doppler waveform between 24-30 weeks of pregnancy. MAIN OUTCOME MEASURES: Preeclampsia (PET), small-for gestational-age at birth (SGA), preterm delivery (PTD), perinatal death. RESULTS: Two hundred thirty-two women (70.3%) had a normal uterine artery Doppler waveforms, and 98 (29.7%) demonstrated either unilateral or bilateral notching. In women where notching was present, 20 (20%) developed PET compared with 8 (3.5%) in the normal group [Odds ratio (OR) 7.2, CI 3-17]; SGA birthweight was present in 24 (24.5%) of the notched group and in 21 (9%) of normal group (OR 3.3; CI 1.7-6.2); 40 (41%) of the notched group delivered preterm ( < 37 weeks) as compared with 37 (16%) of the normal group (OR 7.9; CI 4.6-13). This difference was even more marked when delivery before 32 weeks was considered, occurring in 8 (8%) of the notched group and 4 (1.7%) of the normal group (OR 11.5; CI 4.5-29.4). Of the six perinatal deaths, five (5.1%) occurred in the notched group (OR 12.4; CI 1.4-108). CONCLUSION: This study demonstrates that the addition of uterine Doppler waveform analysis to the monitoring profile of women at risk of PET, SGA, IUD and preterm delivery can further define those in a higher risk group and the majority that have a risk no higher than the background. PMID- 16036399 TI - Oxidant-antioxidant system changes relative to placental-umbilical pathology in patients with preeclampsia. AB - OBJECTIVE: It is speculated that lipid peroxidation is responsible for the pathologic changes that occur in the uteroplacental vasculature of women with preeclampsia. The aim was to investigate this proposed relationship. MATERIALS AND METHODS: The prospective study involved 90 pregnant women. Thirty had mild preeclampsia, 30 had severe preeclampsia, and 30 were healthy pregnant women (controls). The data collected for each case were umbilical cord and placental pathologies, plasma malondialdehyde (MDA) level, and levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity in erythrocytes. Group findings were compared. RESULTS: The mean MDA level in the severe preeclampsia group was higher than the corresponding findings in the mild preeclampsia and control groups (p < 0.001 for both). Also, the MDA level in the mild preeclampsia group was significantly higher than was the control level (p < 0.001). The mean SOD activity level in the severe preeclampsia group was lower than the corresponding results in the mild preeclampsia and control groups (p < 0.001 for both). The mean GSH-Px levels in the mild and severe preeclampsia groups were both significantly lower than was the corresponding finding in the control group (p < 0.01). Compared to the control group, both preeclampsia groups had significantly higher frequencies for placental infarction, villous fibrosis, increased numbers of syncytial nodes, and thickening of vessel walls and lumen obliteration (p < 0.001 for all). Villous fibrinoid necrosis, perivillous fibrosis, and increased villous vascularization were also significantly more frequent in both preeclampsia groups than in the control group, but the differences for these parameters were smaller (p < 0.01 for all). Examination of the samples from the placental ends of the umbilical cords revealed significantly higher frequencies of endothelial irregularity, endothelial shedding, and basal membrane thickening in both preeclampsia groups than in those of the control group (p < 0.001). The same findings were noted in the middle sections of the cords (p < 0.001). At the fetal ends of the umbilical cords, both preeclampsia groups had higher frequencies of endothelial irregularity than did the control group (p < 0.001); however, the frequencies of the more severe pathologic findings (endothelial shedding, basal membrane thickening) in the three groups were similar. CONCLUSION: The frequencies of pathologic changes in the placenta and umbilical vessels of women with preeclampsia parallel the severity of this condition. These changes also parallel plasma levels of MDA, the end product of lipid peroxidation. PMID- 16036400 TI - Second pregnancy outcomes following preeclampsia in a first pregnancy. AB - OBJECTIVES: We compared second pregnancy outcomes among women with and without preeclampsia in their first pregnancies who all had second pregnancies without preeclampsia. METHODS: One hundred thirty women with and 6148 without preeclampsia in their first pregnancies, who all had nonpreeclamptic second pregnancies, were included. Outcomes, including delivery gestational age, birthweight, small-for-gestational-age (SGA), and preterm delivery were compared. RESULTS: Overall, second pregnancy outcomes were not different between women with and without preeclampsia in their first pregnancy. However, when women were stratified by gestational timing of preeclampsia, women with preeclampsia at < 34 weeks (N = 22) had smaller infants and delivered earlier in their second nonpreeclamptic pregnancy compared to women with later preeclampsia (N = 108) or no preeclampsia in the first pregnancy. Women with early preeclampsia also had an increased risk of prematurity (< 37 weeks) in second pregnancies that persisted after controlling for confounding factors [Odds ratio (OR = 3.2)]. DISCUSSION: Second, nonpreeclamptic pregnancy outcomes are different between women with previous early preeclampsia and controls but not between late preeclampsia and controls. These findings support other epidemiological data indicating differences between early and late onset preeclampsia as well as a potential relationship of preeclampsia and spontaneous preterm birth. PMID- 16036401 TI - Pulse wave reflection in currently and previously preeclamptic women. AB - OBJECTIVE: Disturbed maternal endothelial function is believed to be central in the pathogenesis of preeclampsia and has been observed to persist for several years following the preeclamptic pregnancy. Endothelial dysfunction has been reported to cause increased pulse wave reflection, a measure of systemic arterial stiffness. This study tested the hypothesis that preeclampsia and a history of preeclampsia are associated with increased pulse wave reflection. DESIGN AND METHODS: We carried out a cross-sectional case-control study of 26 pregnant women with preeclampsia, 26 pregnant controls, 22 normotensive nonpregnant previously preeclamptic women, and 22 nonpregnant controls. Pulse wave reflection was assessed by applanation tonometry on the radial artery. RESULTS: Pregnant preeclamptic women had a significantly higher heart rate-adjusted augmentation index than did pregnant controls (23 +/- 1 vs. 8 +/- 1%, P < 0.001). The augmentation index of women with a history of preeclampsia was similar to that of the nonpregnant controls (9 +/- 2 vs. 9 +/- 2%, P = 0.78). In a multiple linear regression analysis (R2 = 0.76) the augmentation index of pregnant women was independently associated with a diagnosis of preeclampsia (P < 0.001) and heart rate (P < 0.001), but not with mean arterial blood pressure (P = 0.59). CONCLUSIONS: This study demonstrates that pulse wave reflection and, thus, systemic arterial stiffness are increased in pregnant women with preeclampsia, but not in normotensive nonpregnant women with a history of preeclampsia. The results support the concept of generalized vascular dysfunction in preeclampsia. PMID- 16036402 TI - The incidence of severe complications of preeclampsia. AB - OBJECTIVES: To examine the incidence of life-threatening (near-miss) complications, including hemolysis, elevated liver enzymes and low platelets (HELLP) syndrome occurring in women with preeclampsia in the Grampian region of Scotland between 1981 and 2000. SUBJECTS AND METHODS: All women who were resident in a geographically determined region (the Grampian region of Scotland) and who developed moderate to severe preeclampsia in the time period 1981 to 2000 were identified from the Aberdeen Maternity and Neonatal Databank. All complications occurring antepartum, intrapartum, or postpartum in these pregnancies were listed as International Classification of Diseases (ICD) codes and significant complications were identified from these. The cases of HELLP syndrome were identified by case note review. Data were analyzed using Statistical Package for Social Scientists (SPSS). RESULTS: A total of 4188 cases of preeclampsia were identified over the 20-year period. Six percent (approx. 1 in 16) of preeclamptics developed one or more major systemic complications. The incidence of placental abruption and eclampsia was 2.8% and 1.65%, respectively. Hematological complications were most common, with reduced platelets accounting for about half of these cases; although, a definite diagnosis of HELLP syndrome could only be made in 13 cases over 20 years. After adjusting for gestational age, a woman was 1.14 times more likely to have a caesarean delivery (95% C.I. 1.08, 1.20) if she had complicated preeclampsia. She was also more likely to have a stillbirth [Odds Ratio (O.R.) = 1.45 (95% C.I. (confidence interval) 1.02, 2.29)] or a neonatal death [O.R. = 2.25 (95% C.I. 1.12, 4.260]. CONCLUSIONS: There has been a gradual decline in the rate of preeclampsia in the Grampian region of Scotland over the time period 1986 to 2000; although, the percentage of pre-eclamptics who developed one or more complications has not decreased appreciably. In fact, the incidence of eclampsia and placental abruption has increased in the most recent five years. The presence of complications in preeclampsia is associated with a poor neonatal outcome. PMID- 16036403 TI - Raised leptin concentrations in feto-placental tissues from women with preeclampsia. AB - OBJECTIVE: The aim of this study was to estimate the levels of leptin in the amnion, chorion laeve, and placenta and to examine for any differences in leptin levels in these tissues from preeclamptic and normotensive pregnant women. METHOD: Fresh samples of amnion, chorion laeve, and placenta were obtained from 10 normotensive and 10 preeclamptic women following vaginal delivery. After repeated washing with Krebs Heinsleit solution, 2 g of each tissue were homogenized separately. Following centrifugation, the supernatant was removed and stored at -80 degrees C for analysis of leptin at a later date. Leptin in supernatants from homogenates was measured using commercially available radioimmunoassay (RIA) Kits. Statistical analysis was performed using student's t test for independent samples and all results are expressed as mean +/- SEM. RESULTS: Leptin concentrations were significantly higher in all the three tissues from preeclamptic women when compared to the corresponding tissues from normotensive controls (0.52 +/- 0.1 vs. 1.15 +/- 0.41; 0.99 +/- 0.16 vs. 2.19 +/- 0.71; and 10.28 +/- 1.06 vs. 13.95 +/- 1.13 in the amnion, chorion laeve, and placenta, respectively, from normotensive pregnant women and women with preeclampsia; p < 0.05). CONCLUSION: Leptin concentration is significantly raised in fetoplacental tissues from women with preeclampsia. Its role in preeclampsia remains unclear and it is not possible to say from this study if the raised levels of leptin evident in placentas from preeclamptic women are a cause or consequence of preeclampsia, although the latter seems more likely. PMID- 16036405 TI - Dietary omega-3 polyunsaturated fatty acid supplementation and airway hyperresponsiveness in asthma. AB - Asthma prevalence continues to increase despite the progress that has been made in the treatment options for asthma. Alternative treatment therapies that reduce the dose requirements of pharmacological interventions would be beneficial, and could potentially reduce the public health burden of this disease. There is accumulating evidence that dietary modification has potential to influence the severity of asthma and reduce the prevalence and incidence of this condition. A possible contributing factor to the increased incidence of asthma in Western societies may the consumption of a pro-inflammatory diet. In the typical Western diet, 20-25-fold more omega (n)-6 polyunsaturated fatty acids (PUFA) than n-3 PUFA are consumed, which results in the release of pro-inflammatory arachidonic acid metabolites. Eicosapentaenoic acid and docosahexaenoic acid are n-3 PUFA derived from fish oil that competitively inhibit n-6 PUFA arachidonic acid (AA) metabolism and this reduce the generation of pro-inflammatory 4-series leukotrienes (LTs) and 2-series prostaglandins (PGs) and production of cytokines from inflammatory cells. These data are consistent with the proposed pathway by which dietary intake of n-3 PUFA modulates lung disease. This article will review the existing information concerning the relationship between n-3 PUFA supplementation and airway hyperresponsiveness in asthma. It includes studies assessing the efficacy of n-3 PUFA supplementation in exercise-induced bronchoconstriction. This review will also address the question as to whether supplementing the diet with n-3 PUFA represents a viable alternative treatment regimen for asthma. PMID- 16036406 TI - Intensive care unit admission for asthma: a marker for severe disease. AB - BACKGROUND: There appears to be an asthma epidemic in the United States and other developed countries, with increasing prevalence and severity of asthma. Despite these trends, the understanding of severe asthma remains limited. Research has been hampered by the lack of clear methodology for identifying a cohort of adults with severe asthma. We systematically evaluated a method for defining a cohort of adults with severe asthma based on recent intensive care unit admissions for asthma. METHODS: We used survey interview and computerized utilization data from a cohort of 400 adults with severe asthma who were enrolled after hospitalization at 17 Northern California Kaiser Permanente hospitals. To assess asthma severity, we used a multifaceted approach that combined structured telephone interview data with computerized utilization data. Using a referent group of adults who were hospitalized without intensive care unit (ICU) admission (n=282), we examined whether ICU admission is a valid marker for severe disease (n=118). RESULTS: Adults with asthma who had recent ICU admission had greater severity-of-asthma scores, controlling for sociodemographic factors, smoking, and atopic history (mean score increment 2.3 points; 95% CI 1.3 to 3.2). The ICU subjects also had poorer asthma-specific quality of life (mean score increment 6.6 points; 95% CI 3.5 to 9.8) and were more likely to indicate severe self-perceived asthma (49% vs. 22%, p<0.0001). Adults with recent ICU admission were more likely to have seen an asthma specialist during the past year (27% vs. 16%). They were also more likely to have had an asthma-related emergency department visit (37% vs. 26%), hospitalization (17% vs. 6%), and ICU admission during the previous year (5% vs. 0.7%) (p<0.05 in all cases). Among the 311 adults with asthma who had continuous Kaiser Permanente pharmacy benefits for the previous 12 months, a greater proportion of the ICU group received inhaled corticosteroids during the 12 months prior to hospitalization (78% vs. 65%, p=0.024). CONCLUSIONS: Admission to the ICU for asthma identifies a subgroup with severe disease, providing a valid methodology for defining a cohort of adults with severe asthma. PMID- 16036407 TI - Characterization of a population of monozygotic twins with asthma. AB - BACKGROUND: Asthma is a complex inflammatory lung disease that results from allergic and nonallergic environmental exposures in genetically susceptible individuals. The interrelationship and impact of environmental and genetic determinants on the development of asthma is still unclear. Monozygotic twins (MZT) have been considered a potentially useful population to investigate the role of genetic and environmental factors on asthma because of their identical genetic background, similar childhood environmental exposures, and expected high concordance rate for asthma. METHODS: A detailed questionnaire designed to obtain demographic information, medical history/severity of asthma, and environmental exposures was distributed to 43 pairs of MZT pairs where one or both probands had a history of asthma. All twin pairs were clinically evaluated for atopy by skin testing. Asthma was assessed by questionnaire, baseline spirometry, disease severity, airway reactivity, and medication scores. RESULTS: Based on a history of physician-diagnosed asthma, disease severity, airway reactivity, and medication scores, 22 MZT pairs were identified as concordant (C) for asthma while 21 MZT pairs were discordant (D) for asthma. Those MZT-C for asthma were on average older and more often female. Adjusted odds ratios for age and gender revealed that MZT-C for asthma had a 2.3 greater likelihood of childhood tobacco exposure and a 1.8 greater likelihood of early exposure to a pet compared to MZT D pairs. In addition, MZT-C for asthma had a 2.4 greater likelihood of being full term vs. premature at birth. None of these odds ratios were statistically significant due to a small sample size. However, a trend for a greater likelihood of earlier environmental exposures was observed for MZT-C asthma pairs. Among MZT D probands, a history of asthma in one proband was supported by a lower FEV1 and higher airway reactivity, disease severity, asthma symptom, and medication scores compared to the nonasthma proband. There was significant intrapair heterogeneity among MZT-C and MZT-D probands in their positive skin test responses to specific aeroallergens. CONCLUSION: Cross-sectional evaluation of larger MZT populations where one or both proband has asthma with a long-term prospective follow-up may be a feasible way for identifying the impact of environmental determinants as independent risk factors for the development of asthma. PMID- 16036408 TI - Local ecological factors, ultrafine particulate concentrations, and asthma prevalence rates in Buffalo, New York, neighborhoods. AB - Previous to this study various healthcare utilization studies and house-to-house surveys had shown that Buffalo's west side had a high utilization rate for asthma and high asthma prevalence in comparison with neighboring communities. The relative contributions of traffic-related pollution and personal and local ecological factors to the high asthma rates were still unknown. To investigate the potential roles of personal home environmental factors and local ecological factors in variations of asthma prevalence in Buffalo neighborhoods, we conducted a cross-sectional survey of a systematic random sample of 2000 households in the city of Buffalo, New York, with a response rate of 80.4%. We found that the odds of having at least one person with asthma per household on Buffalo's west side was 2.57 times [95% confidence interval (CI)=1.85-3.57] that of Buffalo's east side. There were no statistically significant differences in the odds of finding at least one person with asthma in households of other Buffalo neighborhoods. We further found no difference in the odds of having asthma on Buffalo's west side even after correcting for race/ethnicity, household triggers of asthma, and socioeconomic factors. Monitoring ultrafine particulates showed increased levels in communities downwind of the Peace Bridge Complex and major roadways supplying it. A multiple-regression model showed that asthma prevalence may be influenced by humidity and ultrafine particulate concentrations. These results suggest that increased asthma risk may be influenced by chronic exposure to personal and local ecological factors. PMID- 16036409 TI - Parental smoking and smoking behavior in asthmatic and nonasthmatic adolescents. AB - BACKGROUND: Smoking may have serious consequences for asthmatics. Nevertheless, studies have shown that smoking behavior among asthmatics is similar to or even higher than that of nonasthmatics. Since the relationship between parental smoking and child smoking is well established, this study examined whether the association between parental and child smoking behavior is similar for asthmatic and nonasthmatic adolescents. The impact of parental smoking history was also explored. METHODOLOGY: A cross-sectional sample of 10,087 Dutch adolescents was used to assess the prevalence of asthma and smoking behavior. Moreover, respondents had to report whether one of their parents currently smoked, had stopped, or had never smoked. In case a parent was a former smoker, the respondent had to report his or her own age at the moment that the parent stopped smoking. RESULTS: Logistic regression analyses showed that, compared with nonasthmatic adolescents, asthmatic adolescents were more likely to have smoking parents. Furthermore, similar associations were found between parental smoking and adolescent smoking among asthmatic and nonasthmatic adolescents. The time at which maternal smoking ceased was associated with a decreased likelihood for ever smoking for both asthmatic and nonasthmatic adolescents. CONCLUSIONS: Asthmatic adolescents need to become more aware of the health risks of smoking. Therefore, tailor-made antismoking campaigns are needed at schools to reduce misconceptions among asthmatic adolescents about the risks of smoking. In addition, a personal intervention approach aimed particularly at smoking parents of an asthmatic child, may make them aware of the consequences for their offspring and help them to stop smoking. PMID- 16036410 TI - Low prevalence of asthma in westernizing countries-myth or reality? Prevalence of asthma in Estonia--a report from the "FinEsS" study. AB - AIM: The aim of this study was to examine the prevalence of asthma among adults in Estonia by using different diagnostic methods and criteria for the disease. METHODS: In 1995-1996, a postal questionnaire was randomly distributed in three areas of Estonia to a representative sample of 22,579 subjects aged 15-64 years. The participation rate was 78%. A clinical follow-up study including structured interview, skin prick test, lung function, and methacholine test, was performed from 1997 to 2000 among randomly selected responders to the postal questionnaire. Of 2676 subjects, 53% participated. RESULTS: The prevalence of physician diagnosed asthma for the postal questionnaire (PQ) was 2.7%, and in the same subjects for the structured interview (SI) 3.8%. Respiratory symptoms, except recurrent wheeze, were more common in the SI than PQ. Combinations of symptoms, except wheezing with breathlessness apart from cold, were more prevalent in the PQ responders. The prevalence of asthma defined by different symptom combinations varied from 5.4% to 8.2%. Among responders, 71-87% demonstrated bronchial hyperreactivity defined as methacholine reactivity=8 mg/mL. The symptom combinations used as surrogate variables for asthma were strongly associated with a positive skin test. CONCLUSION: The low prevalence of physician-diagnosed asthma probably reflects a considerable underdiagnosis of asthma in Estonia. Disease criteria for asthma based on symptom combinations together with hyperreactivity yielded a prevalence of 5%-8%, which is similar to the prevalence of asthma among adults in neighboring Western countries. PMID- 16036411 TI - Acute bronchodilator response has limited value in differentiating bronchial asthma from COPD. AB - BACKGROUND: Acute responsiveness to inhaled bronchodilators is often used to differentiate between bronchial asthma and chronic obstructive pulmonary disease (COPD). The response can be expressed in terms of a change in FEV1 and FVC in several ways-as absolute change, change as percent of baseline value, or as percent of predicted value with different thresholds for a positive test. A comprehensive evaluation of the diagnostic value of these different methods of expressing the acute bronchodilator response has not been carried out. METHODOLOGY: Response to inhaled salbutamol was measured by spirometry in 200 asthmatics and 154 patients with COPD. The sensitivity, specificity, and positive and negative predictive values of different methods of expressing responsiveness were calculated. Receiver operative characteristic curves were obtained. RESULTS: None of the expressions of response gave a clear-cut separation between the two diseases. A DeltaFEV1>or=0.2 L gave the most satisfactory combination of sensitivity (73%) and specificity (80%) and the highest positive (82%) and negative predictive values (69%) for diagnosing asthma. These values were superior to those obtained for the ERS or the ATS criteria for reversibility (DeltaFEV1% predicted >or=9% and DeltaFEV1 of >or= than 12% and 0.2 L over the baseline, respectively), which had almost similar diagnostic characteristics. This was confirmed by the area under curve of the ROC plots. Expressions of response in terms of changes in FVC were unsatisfactory in separating the two diseases. CONCLUSIONS: It was concluded that the test of acute bronchodilator responsiveness has limited diagnostic value in separating asthma and COPD. PMID- 16036412 TI - Recent trends in asthma hospitalization and mortality in the United States. AB - OBJECTIVE: To study the recent trends in asthma hospitalization and mortality rates by age, gender, and race categories in the United States. METHODS: The National Hospital Discharge Survey Database for the years 1995 to 2002 was used to examine trends in asthma hospitalization. An International Classification of Diseases Ninth Revision, Clinical Modification (ICD-9-CM) code of 493.0 was used to identify primary hospitalization for asthma. Hospitalization rates were estimated using U.S. Census Bureau population estimates as denominators. Mortality data was obtained from the Center for Disease Control and Prevention Mortality Database. Death from asthma was identified using ICD-9-CM codes (493.0) for the years between 1995 and 1998 and ICD-10 codes (J45-J45.9) for 1999 and afterwards. Asthma hospitalization and mortality rates were estimated per 10,000 and per 100,000 populations, respectively. Crude relative risks (RR) were estimated to compare risks between various groups. RESULTS: During the study period the age-adjusted asthma hospitalization rate decreased by 16.3% among white females (from 13.4/10,000 in 1995-1996 to 11.2/10,000 in 2001-2002), and by 7% (from 8.14/10,000 in 1995-1996 to 7.56/10,000 in 2001-2002) among white males. Among blacks the decrease in hospitalization rate was by 13.9% (from 38.18/10,000 in 1995-1996 to 32.86/10,000 in 2001-2002) in males and by 14.4% (from 40.21/10,000 in 1995-1996 to 34.42/10,000 in 2001-2002) in females. A narrowing of the black to white disparity in asthma hospitalization rate was noted for children younger than 10 years of age. On the other hand, the racial disparity among subjects 10 years and older narrowed until 2000 but has started to widen since then. The overall decrease in asthma mortality rate was evident for the age group=5, but remained unchanged for the age group less than five. The age adjusted asthma mortality rate has also decreased by 22.2% in blacks (from 3.33/100,000 in 1995 to 2.59/100,000 in 2001) and by 38.4% in whites (from 1.26/100,000 in 1995 to 0.78/100,000 in 2001). CONCLUSION: This study confirms that both asthma hospitalization and mortality rates decreased during the study period and the black to white racial disparity in asthma hospitalization has narrowed for children younger than 10 years of age. For those subjects 10 years and older the racial disparity in hospitalizations narrowed until 2000 but started to widen since then. The widening racial gap in adults is disconcerting and needs further observation to assess its persistence. PMID- 16036413 TI - The relationship of caretaker anxiety to children's asthma morbidity and acute care utilization. AB - OBJECTIVES: This study examined self-reported psychological distress in caretakers of inner-city children with asthma and examined its association to disease severity, as measured by day and night symptoms, and to acute care utilization for asthma. METHOD: We enrolled 193 English and Spanish-speaking caretakers (86% were mothers) of 2- to 12-year-olds who had at least one asthma hospitalization at a large urban medical center. They completed an interview asking about children's asthma symptoms and acute care utilization (provider visits, emergency department visits, hospitalizations for asthma) and a 29-item psychiatric symptom inventory. National guidelines criteria were used to classify asthma severity into three categories based on caretaker report of day and night symptoms in the past 4 weeks. These were: moderate-severe persistent (37%), mild persistent (27%), and mild intermittent (35%) asthma. RESULTS: Caretakers of children who were reported as having moderate-severe persistent asthma symptoms exhibited higher anxiety than caretakers reporting milder symptoms in their children, but symptom severity was unrelated to depression, anger, cognitive disturbance, or overall distress in caretakers. Three measures of acute care utilization in the last 6 months increased with reported symptom severity, but they were not associated with caretaker distress. CONCLUSIONS: Providers treating children with asthma should consider the potential importance of caretaker reports of daily symptoms both for the child's physical functioning and for parental anxiety. PMID- 16036414 TI - Treatment persistence with leukotriene receptor antagonists and inhaled corticosteroids. AB - BACKGROUND: Leukotriene receptor antagonists (LTRAs) and inhaled corticosteroids (ICSs) must be taken continuously to control persistent asthma. We compared the use of LTRAs and ICSs in patients with similar level of asthma control at treatment initiation with particular attention to treatment persistence. METHODS: Two cohorts of 15 to 45 year old patients with asthma were selected from the Quebec Health Insurance Plan Database between January 1, 1998, and December 31, 2000. We first identified new users of LTRAs and from the remaining patients, we selected new users of ICSs. The ICS patients were then one-to-one matched to LTRA patients on the use of short-acting beta2-agonists and oral corticosteroids in the year prior to the date of the first LTRA or ICS dispensation (index date). We compared compliance to initial therapies using Cox proportional hazards models. RESULTS: Each of the LTRA and ICS cohorts included 2200 patients. Multivariate model showed that compliance was significantly better for LTRAs than for ICSs [adjusted rate ratio of treatment discontinuation (aRR), 0.46; 95% confidence interval (CI), 0.42-0.49]. If in both groups all medications filled were taken at the prescribed dose, the annual percent of days on therapy for LTRA users would have been twice that for ICS users (38% vs. 19%; p<0.0001). CONCLUSION: The findings of this observational study indicate a far from optimal persistence to LTRAs and ICSs in asthmatic patients. The superior persistence to LTRAs might result in better effectiveness. PMID- 16036415 TI - Eosinophils from asthmatics release IL-5 in an autocrine fashion to prevent apoptosis through upregulation of Bcl-2 expression. AB - Interleukin (IL)-5 plays an important role in maintaining the survival of eosinophils via the specific alpha-subunit of its receptor. Apoptosis, a form of programmed cell death, is thought to represent a mechanism that promotes the resolution of eosinophilic inflammation in asthma. The aim of our present study is to investigate whether IL-5 acts in an autocrine fashion on eosinophil apoptosis in asthmatics. Immunoreactivities of intracellular IL-5 and IL-5 receptor alpha-subunit (Ralpha) were detected uniquely on the eosinophils. The magnitude of IL-5 and IL-5 Ralpha expression on eosinophils was significantly higher in asthmatics than that of normal subjects (p<0.05) determined by flow cytometry. Apoptosis of eosinophils was measured by the propidium iodide staining method and DNA ladder. The percent of apoptotic eosinophils from asthmatics was significantly increased by coincubation with anti-hIL-5 Ralpha Ab (0.1, 0.5, and 2.5 microg/mL) for 1, 2, or 16 hours than was those of corresponding controls (p<0.05, n=8). However, there was no significant effect of anti-hIL-5 Ralpha Ab on eosinophil apoptosis in normal subjects. Furthermore, the expression of B-cell lymphoma-2 (Bcl-2) proteins was significantly inhibited by the anti-hIL-5 Ralpha Ab or antisense IL-5 oligonucleotides in asthmatics (p<0.05, n=8), but there was no significant change in eosinophils from normal subjects. This study demonstrates that eosinophils from asthmatics release IL-5 in an autocrine fashion to act on their own IL-5 receptors in prevention of apoptosis through the upregulation of Bcl-2 expression. PMID- 16036419 TI - New ideas for therapy in ALS. PMID- 16036420 TI - Report from the Italian Ministerial Committee regarding the diagnosis, care and assistance of patients with ALS. PMID- 16036421 TI - Primary lateral sclerosis, hereditary spastic paraplegia and amyotrophic lateral sclerosis: discrete entities or spectrum? AB - Among the motor neuron diseases, three share the clinical features of prominent upper motor neuron signs--amyotrophic lateral sclerosis (ALS), primary lateral sclerosis (PLS) and the hereditary spastic paraplegias (HSP). While genetic testing can assist in the identification of several variants of the latter, in the remaining cases, including those in which spasticity may be associated with amyotrophy, clinical differentiation of the three disorders may prove difficult. In this paper we review the evidence that these are distinct disorders and conclude that, for ALS and PLS particularly, there may be justification in considering them as single points along a continuum of multisystem disorders with conspicuous motor neuron involvement. Only through the development and application of exacting clinical diagnostic criteria to epidemiological studies, along with greater numbers of post-mortem examinations, however, will these questions be answered fully. PMID- 16036422 TI - Neurophysiological measures in amyotrophic lateral sclerosis: markers of progression in clinical trials. AB - In this review we evaluate clinical neurophysiological methods, originally described for use in diagnosis that can be applied to measurement of change during the progress of amyotrophic lateral sclerosis (ALS). Such measurements are potentially important in clinical trials, and also in clinical practice. We have assessed methods for lower and upper motor neuron function, including conventional EMG, nerve conduction and F-wave studies, the derived Neurophysiological Index, motor unit counting methods (MUNE), and transcranial magnetic motor cortex stimulation. We have also addressed the validity of measurements of electromechanical coupling. Methods for measuring muscle strength are beyond the scope of this review. We conclude that MUNE, M-wave amplitude and the Neurophysiological Index are sufficiently reliable, sensitive, and relevant to the clinical problem of ALS, to be used in clinical trials in the disease. Transcranial magnetic stimulation is of limited value, but a combination of the measurements made as part of this technique may also be useful. We conclude that clinical neurophysiological techniques should now be used in measuring change in clinical trials in ALS. PMID- 16036423 TI - Identification of candidate drugs for the treatment of ALS. AB - A consortium of investigators interested in neurodegenerative diseases collaborated to screen 1040 drugs in multiple neurodegenerative disease assays. One model of amyotrophic lateral sclerosis (ALS) pathogenesis in particular incorporated glutamate exposure in enriched primary rat motor neuron cultures. In this model 78 compounds decreased motor neuron death caused by 100 microM glutamate. Almost all these pharmacological agents act at one or more of the following cellular targets: 1) protein synthesis inhibition; 2) Cox inhibition; 3) regulation of anion flux; 4) modulation of GABA receptors; 5) antioxidant, and 6) cell cycle inhibition. The most prevalent mode of action was the regulation of intracellular calcium. These data extend the understanding of motor neuron degeneration and identify a number of cellular targets for the improvement of combined therapies for neurodegenerative disease. PMID- 16036424 TI - Prognostic factors for survival in amyotrophic lateral sclerosis patients treated with riluzole. AB - The objective of this study was to identify prognostic factors for survival in amyotrophic lateral sclerosis from a large prospective observational study performed in France. The study included a cohort of 2069 patients fulfilling broad entry criteria treated with riluzole. Over 100 demographic, biological, clinical and quality-of-life variables were monitored and assessed for their effect on survival. Patients were randomized post hoc into two groups: one group (two-thirds of the patients) to generate the prognostic models and one group (one third of the patients) to validate the resulting models. Thirteen variables were found to affect survival independently and were used to construct a survival prediction score, RL401. These included age, disease duration, slow vital capacity, intensity of tiredness (visual analogue scale), number of body levels with spasticity, atrophy and/or fasciculations, cough, distal muscle strength, household income, depression and two biological parameters, plasma creatinine levels and neutrophil counts. A simplified score, RL401S, was constructed, designed to be easy to use and interpret. The predictive powers of the two scores were similar. PMID- 16036425 TI - The IVS1 +319 t>a of SOD1 gene is not an ALS causing mutation. AB - Amyotrophic lateral sclerosis (ALS) is caused by mutations in the gene for Cu/Zn superoxide dismutase (SOD1) in 10% of familial and sporadic cases. During the SOD1 analysis of 9 FALS and 121 SALS, in only one sporadic case we found the exonic mutation N19S; in 15 SALS patients we found a 319t>a variation in IVS1 sequence, at 108 bp upstream from exon 2. This variation has an unusually high frequency of 11% and is always in linkage disequilibrium with a described polymorphism in IVS3, +34a>c. The 319t>a variation is classified in two different public databases, HGMD and The ALS Online Database, as a splicing mutation and not as a polymorphism. The unusually high frequency of this mutation in our patients prompted us to determinate its frequency in 130 age- and gender- matched healthy controls and in 54 patients with Alzheimer's disease. We found again linkage disequilibrium with the polymorphism in intron 3, and the frequency of 11% and 7.8%, respectively. These results strongly support the idea that the IVS1 +319 t>a alone is not an ALS causing mutation, and that special care must be taken in the interpretation of data from mutations databases for correct genetic counselling. PMID- 16036426 TI - Rating the severity of ALS by caregivers over the telephone using the ALSFRS-R. AB - We tested the hypothesis that the ALSFRS-R can be accurately administered exclusively to the spouse/caregiver over the telephone (phone) in 31 patients with clinically possible, probable, or definite ALS (El Escorial criteria) by comparing the phone response to the consensus assessment during a clinic visit using a standardized script. We also compared the responses of the spouse/caregiver, during a private assessment, to the consensus assessment. Both the phone and private assessments were highly correlated with the consensus assessment, even considering that the evaluations were performed by multiple phone assessors and, in three instances, multiple home assessors. These results suggest the possibility of using this approach to conduct research in ALS. PMID- 16036427 TI - Optimal methods to characterize the G93A mouse model of ALS. AB - In the present study, we used the SOD1 (G93A) mutant transgenic mice as a model of amyotrophic lateral sclerosis (ALS). This model is widely used as a laboratory tool to study experimental treatments in vivo for ALS to investigate new therapeutic strategies for this neurodegenerative disease. Such studies require the objective quantification of different parameters while mice develop the disease. We have applied a battery of different and specific tests: scoring of motor deficits by a trained observer, weighing, survival measure, hanging wire test, rotarod task and electromyography, most of them commonly used to evaluate G93A animals. We have critically compared these methods, showing the significant influence of gender on the onset of symptoms, and the optimal moment to apply each test. These results should be taken into account in future therapeutic assays on this ALS model. PMID- 16036429 TI - Primary lateral sclerosis, hereditary spastic paraplegia, and mutations in the alsin gene: historical background for the first International Conference. PMID- 16036430 TI - Sporadic amyotrophic lateral sclerosis: a hypothesis of persistent (non-lytic) enteroviral infection. AB - Because of recently reported reverse transcriptase polymerase chain reaction evidence of enterovirus in sporadic amyotrophic lateral sclerosis (SALS) and because of newly available anti-enteroviral drugs binding enteroviral capsids, it is reasonable to re-formulate an enteroviral hypothesis of SALS using recent advances in molecular virology. Viral persistence is non-lytic and non-cytopathic infection that evades host's immune surveillance. Enteroviruses are known to cause persistent as well as lytic infection both in vitro and in vivo. Both virion as well as host factors modulate between persistent and lytic infection. Apoptosis, or programmed cell death, is a process of active non-necrotic cell death. It has complex interplay with viruses and may be either promoted or opposed by them. Apoptosis is a major factor in motor neuron death in SALS. Viral tropism is the process by which viruses select and propagate to target cells. It is controlled by capsid conformation and surface receptors on host cells. Enteroviruses have a region on their capsids known as the canyon which docks on such receptors. Docking induces conformational changes of the capsid and genome release. Poliovirus, tropic for motor neurons, docks on the poliovirus receptor, about which much is known. The virus penetrates the motor system focally after crossing either the blood-muscle or the blood-brain barriers. It propagates bidirectionally along axons and synapses to contiguous motor neurons, upper as well as lower, which sequester infection and create avenues for spread over long distances. If chronic and persistent rather than acute and lytic, such viruses trafficking in a finite system of non-dividing cells and inducing apoptosis would cause cell death that summates linearly rather than exponentially. Taken together, these explain signature clinical features of SALS - focal onset weakness, contiguous or regional spread of weakness, confinement to upper and lower motor neurons, and linear rates of progression. The hypothesis predicts the following testable investigations: 1) viral detection may be possible by applying amplification technology to optimally acquired nervous tissue processed by laser microdissection; 2) genetic susceptibility factors such as cell surface receptor polymorphisms may combine with sporadic exposure and chance penetration of the motor system in SALS; 3) a transgenic animal model might be created by inserting such genetic factors into an animal host and inoculating intramuscularly rather than intracerebrally biochemical fractions of SALS motor neurons at vulnerable periods in the developmental life cycle of the transgenic host; and 4) continual long-term administration of anti-enteroviral agents called capsid-binding compounds which stabilize capsids and prevent genome release might be efficacious. PMID- 16036431 TI - Complications and survival following radiologically and endoscopically-guided gastrostomy in patients with amyotrophic lateral sclerosis. AB - In the natural progression of amyotrophic lateral sclerosis (ALS), a state of malnutrition often develops, associated with reduced oral intake, caused by difficulties with swallowing, adequate salivation, and/or anorexia. The placement of a feeding tube should be considered at the point where patients demonstrate swallowing difficulty and/or alteration in nutritional status. The goal of this prospective work was to compare, in 50 patients having definite or probable ALS, the complications after endoscopic (PEG), or radiologically-guided gastrostomy (RIG) and to evaluate their effects on survival. RIG was proposed as first-line therapy when patients had a slow vital capacity (SVC) less than 50% of the theoretical value, or in a case where PEG was refused by the patient. Thirty patients underwent PEG procedure, and 20 had RIG. Among the decisional criteria for placing a gastrostomy tube, risk of aspiration demonstrated by using videofluoroscopy was more frequently observed in the RIG group (65%) than in the PEG group (19.2%) (p=0.002). The two patient populations at time of gastrostomy placement were comparable with respect to age, gender ratio, disease duration before gastrostomy, the form and the gravity of neurological involvement as well as for nutritional criteria. The only significant difference observed between the two groups was the level of the SVC, much lower for patients undergoing RIG: 51.6+/-25.0% versus 67.4+/-26.7% than for the PEG group (p=0.03). The frequency of all complications observed at the moment of placement of gastrostomy tube and during the first month of follow-up was not significantly different between the two groups. Kaplan-Meier survival curves from the date of gastrostomy placement were not different in univariate analysis (p=0.85). In multivariate analysis, survival was not different between one group and the other (p=0.28). The major interest of the RIG technique rests on its greater feasibility and on the possibility of utilizing it in ALS patients who have significant ventilatory compromise. PMID- 16036432 TI - Expression of FKBP12 and ryanodine receptors (RyRs) in the spinal cord of MND patients. AB - We investigated the FKBP12 and ryanodine receptor (RyR) immunoreactivity (IR) in the spinal cords of neurological controls and patients with motor neuron disease (MND). In the neurological controls, the cytoplasm of the spinal anterior horn neurons was stained with anti-FKBP12 antibodies and anti-RyR (type 1 and type 2) antibodies. In the MND cases, the residual neurons in the anterior horn of the spinal cord showed IR for RyR (type 1 and 2) antibodies, while weak IR for anti FKBP12 antibodies was comparable to that of controls. The numbers of neurons recognized with the anti-FKBP 12 or anti-RyR (type 1 and 2) antibodies were counted in the anterior horn of spinal cords from the MND cases and neurological controls. Frequency of neurons stained with anti-FKBP 12 antibody was significantly decreased in the MND cases compared to that in controls (48.7+/ 23.2%, 71.0+/-18.5%, respectively, mean+/-SD, p<0.0005). In the MND cases, numbers of normal-appearing, chromatolytic neurons showing IR to anti-FKBP12 (N19) antibody were significantly decreased compared to those in the controls. Immunoreactivities to anti-RyR antibodies (type 1and 2) in MND cases were present and there was no difference compared to those of the controls. Neurons in the spinal cord anterior horn of Kii-ALS cases with prolonged clinical duration were immunostained with both anti-FKBP12 and anti-RyR (type 1 and 2) antibodies similar to that in the controls. The anterior horn neurons of MND cases of short clinical duration showed absent IR to FKBP 12 antibody but present IR to RyR (type 1 and 2) antibodies. The present result suggests that FKBP12 IR was decreased in the MND cases with short clinical duration. RyR (type 1 and 2) is a major component of the intracellular calcium channel, which mediates calcium induced calcium release. FKBP12, which is an endogenous ligand for RyR, stabilizes the calcium channels preventing calcium leakage in the absence of receptor activation. Imbalance between FKBP12 and RyR IR may play an important role in degeneration due to MND. Further study of the correlation between RyR and FKBP12 should contribute to clarifying the mechanisms of neurodegeneration in MND, including calcium-induced neuronal loss. PMID- 16036433 TI - Autonomic dysfunction in ALS: a preliminary study on the effects of intrathecal BDNF. AB - This pilot study aimed at exploring the effects of intrathecally administered brain derived neurotrophic factor (BDNF) on autonomic functions in patients with ALS. A battery of autonomic sympathetic and parasympathetic tests was performed at baseline and after nine months of treatment in 10 ALS patients participating in a double-blind placebo-controlled phase II/III study of intrathecally administered BDNF. Results of patients treated with BDNF (25 or 150 microg/day) were compared to those receiving placebo. Sudomotor function and blood pressure response to handgrip significantly worsened during the treatment period (55.4+/ 26.1 vs. 38.9+/-23.9 g/m(2)h, p<0.05; 20+/-6 vs. 13+/-4 microHg, p<0.05) whereas other sympathetic and all parasympathetic function tests only tended to be more abnormal at follow-up. Serum norepinephrine levels increased significantly during the nine-months observation period. The results of autonomic function tests were not different between patients treated with BDNF and placebo, but norepinephrine levels were higher in the BDNF group. We conclude that autonomic nervous system function deteriorates along with poorer motor performance independently from treatment with BDNF. The elevation of norepinephrine levels might reflect a non specific up-regulation, and its association with BDNF an autocrine effect. PMID- 16036434 TI - Biochemical characterization of plasma in amyotrophic lateral sclerosis: amino acid and protein composition. AB - In this work, we have studied the amino acid and protein composition of the plasma from a group of 32 ALS patients. As controls, groups of 10 healthy subjects (HC) and 32 patients with other neuromuscular disorders have been analysed. When the HC group was compared with the ALS group there were significant decreases of His (39+/-18 to 24+/-9 microM, p<0.01) and Ala (313+/-62 to 237+/-66 microM, p<0.05), and a significant increase of Asn (89+/-41 to 118+/ 24 microM, p<0.05), for the ALS group. When the three groups were compared, we observed significant decreased concentrations of Ser, His, Thr, Ala, Arg, Tyr, Met, Cys, Ile, and significant increases of Asn, Phe and Lys. An increase of proteolytic products of alpha2-macroglobulin (alpha2-M), an acute-phase serum glycoprotein that functions as a protease inhibitor, has been observed for a subgroup of ALS patients by Western blot. Furthermore, the detection of alpha2-M during disease progression has shown increases of the intact subunit and of a proteolytic product for two of the four patients analysed. Another acute-phase glycoprotein, haptoglobin, which regulates haemoglobin degradation, was not increased for the same group of patients. The results obtained suggested that diet supplementation with His and Ala and modulation of alpha2-M might have some beneficial effects on the course of ALS. PMID- 16036435 TI - The SOD1 transgene in the G93A mouse model of amyotrophic lateral sclerosis lies on distal mouse chromosome 12. AB - The SOD1G93A transgenic mouse strain which carries a human mutant Cu/Zn superoxide dismutase transgene array is a widely studied model of amyotrophic lateral sclerosis. These mice have been used in many breeding experiments to look for interactions with other loci, including transgenic and gene targeted mutations. Therefore, we decided to map the site of the transgene insertion as this may affect the outcome of such breeding experiments. In a fluorescence in situ hybridization experiment we determined that the SOD1G93A transgene insertion site lies on distal mouse chromosome 12. This chromosome also carries the 'Legs at odd angles' locus, which is an entirely unrelated mutation in the dynein heavy chain gene that we have been studying. We have analysed data from a SOD1G93AxLoa cross and determined that the site of the transgene insertion lies proximal of the dynein heavy chain gene on mouse chromosome 12. PMID- 16036436 TI - Screening the metallothionein III gene in sporadic amyotrophic lateral sclerosis. AB - Metallothioneins are proteins involved in antioxidant defence, essential metal homoeostasis and heavy metal detoxification, all mechanisms implicated in sporadic amyotrophic lateral sclerosis (SALS). We therefore looked for changes in the gene for nervous system-specific metallothionein III (MT3) that might explain susceptibility to SALS. DNA was extracted from 87 sporadic ALS and 174 matched controls. The gene for MT3 was sequenced in 20 SALS and 5 control subjects to identify single nucleotide polymorphisms (SNPs). These SNPs were then screened in all subjects. Eight novel SNPs were found in the 5' untranslated region and intron 2 of MT3. No differences were found in the frequency distribution of alleles or haplotypes for these SNPs between the SALS and control groups. The genotype distribution of one SNP (A1422C) was significantly different between ALS and control groups (p<0.02) but this is not likely to be biologically relevant. We conclude that changes in the MT3 gene are unlikely to be responsible for susceptibility to SALS. PMID- 16036437 TI - Serum levels of soluble intercellular adhesion molecule-1 (s-ICAM-1) and soluble endothelial leukocyte adhesion molecule-1(s-ELAM-1) in amyotrophic lateral sclerosis. AB - Immunological disturbances have been implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS). Adhesion molecules are markers of activated endothelial cells up-regulated by action of cytokines. To investigate the activation or inactivation of the vascular cells in ALS, serum soluble intercellular adhesion molecule-1 (s-ICAM-1) and soluble E-selectin (s-ELAM-1) were evaluated (ELISA) in 16 patients with ALS, 30 patients with non-inflammatory neurological diseases (NINDS) and 15 healthy control subjects. Patients with ALS had no higher s-ICAM-1 levels compared with the NINDS patients and the control subjects (p<0.31 and p<0.21, respectively). s-ELAM levels were not statistically significant compared with the NINDS patients and healthy subjects (p<0.21 and p<0.24, respectively). We conclude that the low values of s-ICAM-1 and s-ELAM-1 in the serum of ALS patients do not exclude the presence of immunological abnormality in this disorder. Soluble E-selectin is a glycoprotein which is considered an exclusive marker of endothelial activation. Its low level in our study may suggest a neural rather than an endothelial s-ICAM origin in patients with ALS. PMID- 16036438 TI - Hoarseness due to bilateral vocal cord paralysis as an initial manifestation of familial amyotrophic lateral sclerosis. AB - Bulbar palsy is unusual as an initial manifestation of amyotrophic lateral sclerosis (ALS), although common in the advanced stages. In terms of bulbar palsy as a presenting symptom, dysarthria and dysphagia are of common features. Hoarseness, however, is an initial symptom of ALS in only a small number of patients. We report a 43-year-old female with hoarseness due to bilateral vocal cord paralysis as the first manifestation of ALS. Gene analysis revealed a heterozygous missense mutation in the SOD1 gene, which resulted in an amino acid substitution of isoleucine 149 by threonine. Hoarseness can be the initial symptom of ALS. Therefore, in cases of bilateral vocal cord paralysis of unknown etiology, ALS should be taken into consideration. PMID- 16036439 TI - European ALS Consortium (EALSC): second annual Research Workshop, a summary report. PMID- 16036441 TI - Copy and recall performance of 6-8-year-old children after standard vs. Step-by step administration of the Rey-Osterrieth complex figure. AB - The Rey-Osterrieth Complex Figure (ROCF; Rey 1941; Osterrieth, 1944) is frequently used in the neuropsychological assessment of children and adults. The present study was designed, in part, to examine the impact of providing organizational scaffolding to young children being tested with the ROCF. To this end, 6-, 7-, and 8-year-old children were administered the test either in the standard fashion, or using a format in which the 18 key elements of the figure were introduced sequentially. Participants included 132 children who were randomly assigned to the standard or step-by-step administration groups. Significantly higher accuracy and organization scores for both copy and recall were seen with the step-by-step format than with the standard format, even though children in the step-by-step condition took less time to execute their drawings. Retention of encoded information was not affected by age or testing format. The fact that 6-year-olds in the step-by-step condition performed as well as, or better than, 8-year-olds in the standard condition suggests that the primary problem young children experience with the ROCF lies with organizational strategy formation. Advantages of using the Step-by-step ROCF in clinical practice are discussed. PMID- 16036440 TI - Impaired auditory attention skills following middle-ear infections. AB - The present study investigated auditory attention skills in a sample of children with non-chronic otitis media with effusion (OME). Twenty children with repeated episodes of OME but not found in the need for myringotomy and insertion of ventilating tubes were compared to 20 control children with no known episodes of OME based on parental reports and medical records. Mean age during assessment was 9 years, and none of the children showed signs of impaired language functions and with normal general cognitive abilities. They were assessed with dichotic listening CV-syllables (DLCV-108) free recall and directed attention tasks. The control children showed the expected right ear advantage during free recall and the directed right condition, and demonstrated a shift toward a left ear advantage during the directed left. The children with a history of OME showed a predominant right ear advantage across all three tasks. Although some change in ear accuracy occurred across tasks, impaired auditory attention skills were found following a history of middle ear infections. These results replicate those reported earlier from a sample of children with persistent otitis media with effusion, and suggest that treatment with ventilating tubes does not appear to make any difference in the development of auditory attention skills, whereas occurrence of OME must be considered when testing auditory attentional skills as a part of a neuropsychological assessment. PMID- 16036442 TI - Motor preparation, motor execution, attention, and executive functions in attention deficit/hyperactivity disorder (ADHD). AB - Attention and executive functions were investigated in medicated and unmedicated children with ADHD combined type using a novel selective reaching task. This task involved responding as rapidly as possible to a target while at times having to ignore a distractor. Results indicated that unmedicated children with ADHD showed slow and inaccurate responding. Slow responding reflected problems at the stage of movement preparation but not movement execution. An attentional impairment, rather than a motor planning problem per se, appeared to underlie the slow movement preparation. Inaccurate responding reflected problems with response inhibition and selective attention, impulsivity, set-shifting, and difficulties in maintaining vigilance. Although medicated children with ADHD did not show slow movement preparation, they did show some response inaccuracy, resulting especially from impulsive responding. These findings suggest that ADHD is characterized by slow motor preparation (but not motor execution), and deficits in selective attention, vigilance, and executive functions. Preliminary results suggest that stimulant medication may resolve some of these motor, attentional and executive function deficits. PMID- 16036443 TI - Attentional flexibility and perseveration: developmental aspects in young children. AB - Whereas a growing interest in the development of attentional flexibility (AF) and in perseverative behavior, being one marker of this component, exists in neuropsychological studies and in the domain of developmental psychopathology (e.g., PKU, infantile schizophrenia, autism and Parkinson's disease) (Pennington & Ozonoff, 1996; Stahl & Pry, 2002), only a few studies have concerned themselves with this subject in normal children. It is thus of interest to add more empirical data to the existing literature in this domain. Therefore, the aim of our study was to explore the development of AF and of perseverative errors in young preschool children with normal development, aged 1.5 to 6 years. Using set shifting tasks of increasing difficulty level, three age groups were compared with respect to their AF skills. Results show a developmental factor underlying AF, with different levels of this form becoming more and more complex with age, ranging from a rudimentary visual form to a complex representational form of flexibility. Overall, few perseverative errors occurred and they decreased with age. Results are discussed from a developmental and neuropsychological perspective. PMID- 16036444 TI - Response interference and working memory in 12-year-old children. AB - A group of 69 12-year-old children performed three well-known response interference tasks: the Stroop task, the Eriksen flanker task, and the Simon task. Individual differences in accuracy and speed correlated across the tasks. However, there was no correlation between the interference effects on these three tasks. Stroop interference, but not the Simon or flanker effect, was correlated with working memory capacity, as obtained from the WISC-R. These results may help clarify the nature of ADHD, which is characterized by problems with response interference. PMID- 16036445 TI - Neurobehavioral sequelae of child sexual abuse. AB - This study examined intellectual and memory functioning in a sample of sexually abused children compared to demographically and age-matched controls. The severity of abuse and other pertinent factors were also examined in relation to cognitive performance. Elevated levels of psychopathology were present in the abused children, as well as diminished performance on tasks influenced by attention/concentration. However, after controlling for differences in IQ and socioeconomic status (SES), significant differences in memory function were not found. Results are discussed in the context of stress effects on cognition and the potential resiliency of cognitive function in children undergoing treatment for sexual abuse. PMID- 16036446 TI - Performance on the test of memory malingering in a mixed pediatric sample. AB - Performance on the Test of Memory Malingering (TOMM) was evaluated in a sample of 100 consecutively referred 6 to 16-year-old children with a wide range of clinical diagnoses. In the complete sample, 97 children met actuarially defined criteria for sufficient effort on the TOMM. Two children were correctly identified as providing suboptimal effort and only one case was a possible false positive. Performance on the second trial of the TOMM did not vary with gender, ethnicity, parental occupation, performance on an independent memory test, or length of coma. Although younger children tended to be somewhat less efficient on the TOMM than older children, more than 90% of children in the 6-8 years range met criteria originally developed for adults for sufficient effort on the TOMM. It is concluded that the TOMM is a potentially useful measure of effort in the clinical neuropsychological evaluation of school-age children. PMID- 16036449 TI - Limitations in verbal fluency following heavy burdens of early childhood diarrhea in Brazilian shantytown children. AB - The effects of heavy burdens of diarrhea in the first 2 years of life on specific executive control function like verbal fluency are not well understood. In previous studies, we have shown associations of early childhood diarrhea (ECD) with nonverbal intelligence and school functioning. Therefore, we postulated that ECD might affect early neuropsychological development leading to long-term deficits in normal cognitive development. Based on our extensive 14-year prospective cohort studies of early childhood diarrheal illnesses in a Brazilian shantytown community, we examined ECD correlations between specific impairments of higher mental function and executive skills in shantytown children 5-10 years later (now at 6-12) years of age. Specifically we examined whether heavy diarrheal illnesses correlate with reduced performance on selected tests of executive function. Our study, for the first time, suggests a disproportional impairment in semantic but not phonetic fluency in a subset of children with heavy burdens of diarrhea in their first 2 years of life even when controlling for maternal education, breastfeeding, and child schooling. Similar semantic decrements have been associated with impaired recovery from brain injury. These exploratory studies suggest the importance of verbal fluency tests to assess executive functioning in children challenged by poor nutrition and diarrhea in early life. In addition, our unique findings show the potential influences of early childhood diarrhea on language development that is so critical to productive adulthood and potentially set a foundation for new neuropsychological approaches, which assess early burdens of enteric illnesses on childhood development. PMID- 16036450 TI - Characterization of children's decision making: sensitivity to punishment frequency, not task complexity. AB - On a gambling task that models real-life decision making, children between ages 7 and 12 perform like patients with bilateral lesions of the ventromedial prefrontal cortex (VMPFC), opting for choices that yield high immediate gains in spite of higher future losses (Crone & Van der Molen, 2004). The current study set out to characterize developmental changes in decision making by varying task complexity and punishment frequency. Three age groups (7-9 years, 10-12 years, 13 15 years) performed two versions of a computerized variant of the original Iowa gambling task. Task complexity was manipulated by varying the number of choices participants could make. Punishment frequency was manipulated by varying the frequency of delayed punishment. Results showed a developmental increase in the sensitivity to future consequences, which was present only when the punishment was presented infrequently. These results could not be explained by differential sensitivity to task complexity, hypersensitivity to reward, or failure to switch response set after receiving punishment. There was a general pattern of boys outperforming girls by making more advantageous choices over the course of the task. In conclusion, 7-12-year-old children--like VMPFC patients--appear myopic about the future except when the potential for future punishment is high. PMID- 16036451 TI - Attention problems and executive functions in children with spina bifida and hydrocephalus. AB - This study addressed the incidence of Attention-Deficit/Hyperactivity Disorder (ADHD) subtypes in children with spina bifida meningomyelocele and shunted hydrocephalus (SBH) as well as differences in executive functions among these subtypes. Parent rating scales revealed that 31% of the group with SBH could be identified with AD/HD, mostly the Inattentive type (23%). The group with SBH differed from normal controls on cognitive measures of executive functions, but subtype differences were not significant. Multivariate tests showed that children with SBH were rated with greater difficulties on the Behavior Rating Inventory of Executive Function (BRIEF) compared to controls; those with SBH and any subtype of ADHD differed from those with SBH and no ADHD; and those with ADHD (Combined Type) differed significantly from those with ADHD (Predominantly Inattentive Type). Subtype differences on univariate tests in the latter comparison were significant on the BRIEF Inhibit scale, showing more disinhibition in those with SBH and ADHD (Combined Type), but no significant differences were apparent on the BRIEF Sustain, Shift, and Initiate scales. The results show that the incidence of ADHD in children with SBH exceeds the population rate, is represented by problems with inattention rather than with impulsivity and hyperactivity; and that as with non-brain injured individuals, subtype differences in cognitive function remain to more clearly delineated. PMID- 16036452 TI - Sustained attention and executive functioning performance in attention deficit/hyperactivity disorder. AB - The aim of this study was to further refine the cognitive phenotype of attention deficit/hyperactivity disorder (ADHD), with respect to the ability to sustain attention and executive functioning. Participants were 34 boys with ADHD (combined type) and 28 normal controls. The groups were closely matched for age and IQ. All participants were 12 years of age. Both groups performed a computerized sustained attention task and a response interference task. Measures related to speed, accuracy, and time on task were collected. We found that children with ADHD performed slower, less accurately, more impulsively, and with less stability than controls. Both groups produced more errors with increasing time on task, reflecting reduced vigilance. Importantly, no interaction with time on task was found. The overall pattern of results suggests that measures related to accuracy are more informative than measures related to speed of responding in refining the cognitive phenotype of ADHD. PMID- 16036453 TI - The effect of reinforcement variables on inhibition in children with ADHD. AB - Behavioral inhibition, often cited as a central deficit in children with ADHD, has been shown to change in response to reinforcement. In this preliminary investigation, children with ADHD (n= 20) and matched controls (ages 7 to 12) completed a new version of the stop signal paradigm, the Fire Fighter Game, a measure of inhibition of a prepotent motor response, under three conditions: (1) no reinforcement; (2) immediate reinforcement; and (3) delayed reinforcement. In all conditions, the stop signal reaction time (SSRT) of children with ADHD was consistently longer than controls. Both groups improved significantly with reinforcement, and there was no strong evidence that immediate reinforcement was more effective than delayed reinforcement. However, it appeared that the reason for the changes in SSRT in response to reinforcement differed between the groups. Children in the control group responded faster on go trials, whereas children with ADHD improved their ability to inhibit responding with shorter stop delays. The relevance of these findings is discussed in terms of current theories of ADHD. PMID- 16036454 TI - Repeated assessment of cognition in children and the measurement of performance change. AB - There is limited understanding of the problems associated with repeated neuropsychological assessment in children, including the statistics used to guide decisions about cognitive change. This study investigated the utility of a computerized assessment battery that was specifically designed for the repeated assessment of cognitive function in children. Eighty-seven participants aged 8 to 12 years completed the battery four times within a 2-hour testing session. The results support the application of this assessment battery for measuring cognitive change in children. A novel method for calculating measurement error is employed, and its use in the detection of cognitive change in individual children is discussed. An estimate of the measurement error within each of the tests is provided, and recommendations are made regarding the application of this assessment battery for measuring cognitive change in children. PMID- 16036458 TI - Well-being and fairness in the distribution of scarce health resources. AB - Based on a general thesis regarding the proper resolution of interpersonal conflicts, this paper suggests a normative framework for the distribution of scarce health resources. The proposed thesis includes two basic ideas. First, individual well-being is the fundamental value. Second, interpersonal conflicts affecting well-being should be resolved in light of several conceptions of fairness, reflecting the independent value of persons and the moral significance of responsibility of individuals for the existence of interpersonal conflicts. These ideas are elaborated in several principles that are applied with respect to the distribution of scarce health resources. PMID- 16036459 TI - Enhancement technologies and human identity. AB - As the President's Council on Bioethics emphasized in a recent report, rapid growth of biotechnologies creates increasingly many possibilities for enhancing human traits. This article addresses the claim that enhancement via biotechnology is inherently problematic for reasons pertaining to our identity. After clarifying the concept of enhancement, and providing a framework for understanding human identity, I examine the relationship between enhancement and identity. Then I investigate two identity-related challenges to biotechnological enhancements: (1) the charge of inauthenticity and (2) the charge of violating inviolable core characteristics. My thesis is that a lucid, plausible understanding of human identity largely neutralizes these charges, liberating our thinking from some seductive yet unsound objections to enhancement via biotechnology. PMID- 16036460 TI - Human embryos in the original position? AB - Two different discussions in John Rawls' A Theory of Justice lead naturally to a rather conservative position on the moral status of the human embryo. When discussing paternalism, he claims that the parties in the original position would seek to protect themselves in case they end up as incapacitated or undeveloped human beings when the veil of ignorance is lifted. Since human embryos are examples of such beings, the parties in the original position would seek to protect themselves from their embryonic stages onward. When discussing the basis of equality, Rawls claims that the parties in the original position would guarantee basic rights for all those with the capacity to take part in this original position. To guarantee the basic rights of infants and young children, he goes on to interpret this capacity as a "potentiality that is ordinarily realized in due course." Since human embryos have this potentiality, they too should have basic rights. PMID- 16036461 TI - From the local to the global: bioethics and the concept of culture. AB - Cultural models of health, illness, and moral reasoning are receiving increasing attention in bioethics scholarship. Drawing upon research tools from medical and cultural anthropology, numerous researchers explore cultural variations in attitudes toward truth telling, informed consent, pain relief, and planning for end-of-life care. However, culture should not simply be equated with ethnicity. Rather, the concept of culture can serve as an heuristic device at various levels of analysis. In addition to considering how participation in particular ethnic groups and religious traditions can shape moral reasoning, bioethicists need to consider processes of socialization into professional cultures, organizational cultures, national civic culture, and transnational culture. From the local world of the community clinic or oncology unit to the transnational workings of human rights agencies, attentiveness to the concept of culture can illuminate how patients, family members, and health care providers interpret illness, healing, and moral obligations. PMID- 16036464 TI - Beyond verbal fluency: investigating the long-term effects of bilateral subthalamic (STN) deep brain stimulation (DBS) on language function in two cases. AB - Cognitive functioning has been described as largely impervious to chronic STN-DBS administered over 12-month periods. In relation to the domain of language, however, the effects of STN-DBS are yet to be thoroughly delineated. Verbal fluency tasks represent an almost exclusively applied index of linguistic proficiency relative to neuropsychological research within this population. Comprehensive investigations of the impact of STN-DBS on language function, however, have never been undertaken. The more precise elucidation of the role of the STN in the mediation of language processes, by way of assessments which probe language comprehension and production mechanisms, served as the primary focus of this research. Longitudinal analysis also afforded consideration of the way in which cognitive-linguistic circuits respond to STN-DBS over time. Bilateral STN DBS primarily effected clinically reliable fluctuations (i.e., both improvements and declines) in performance in both subjects on tasks demanding cognitive linguistic flexibility in the formulation and comprehension of complex language. Of particular note, both subjects demonstrated a cumulative increase in the proportion of reliable post-operative improvements achieved over time. The findings of this research lend support to models of subcortical participation in language which endorse a role for the STN, and suggest that bilateral STN-DBS may serve to enhance the proficiency of basal ganglia-thalamocortical linguistic circuits over time. PMID- 16036465 TI - Cerebellar agenesis II: motor and language functions. AB - In a former study of a patient with cerebellar agenesis (HK) mild motor deficits, problems in delay eyeblink conditioning and mild to moderate deficits in IQ, planning behavior, visuospatial abilities, visual memory, and attention were found. The present study reports additional findings in the same patient. In the motor domain, impairments in fine motor manipulations, trace eyeblink conditioning and motor imagination in a functional magnetic resonance (fMRI) study were found. Based on fMRI findings; however, cortical areas involved in a tapping task did not significantly differ from a healthy control group. In the cognitive domain, deficits in speech comprehension as well as verbal learning and declarative memory were present. No significant affective symptoms were observed. Although problems in executive, visuospatial and language tasks are in agreement with the so-called cerebellar cognitive affective syndrome-other possibilities remain. Non-motor impairments in HK might also be a consequence of lacking motor abilities in development and motor deficits may interfere with the performance of parts of the cognitive tasks. In addition, lack of promotion and learning opportunities in childhood may contribute and mental retardation based on extracerebellar dysfunction cannot be excluded. PMID- 16036466 TI - Dissociation of emotional processes in response to visual and olfactory stimuli following frontotemporal damage. AB - Contemporary neuropsychological studies have stressed the widely distributed and multicomponential nature of human affective processes. Here, we examined facial electromyographic (EMG) (zygomaticus and corrugator muscle activity), autonomic (skin conductance and heart rate) and subjective measures of affective valence and arousal in patient TG, a 30 year-old man with left anterior mediotemporal and left orbitofrontal lesions resulting from a traumatic brain injury. Both TG and a normal control group were exposed to hedonically valenced visual and olfactory stimuli. In contrast with control subjects, facial EMG and electrodermal activity in TG did not differentiate among pleasant, unpleasant and neutral pictures. In addition, the controls reacted spontaneously with larger corrugator EMG activity and higher skin conductance to unpleasant odors. By contrast, the subjective feeling states (pleasure and arousal ratings) remained preserved in TG. The covariation between facial and self-report measures of negative valence was also a function of the nature of the olfactory task in the patient only. Taken together, the data suggest a functional dissociation between brain substrates supporting generation of emotion and those supporting representation of emotion. PMID- 16036467 TI - A retrospective journal-based case study of an infant with autism and his twin. AB - This report describes the development of an infant who was later diagnosed with autism, and a direct comparison of his development to that of his twin, from a prenatal period through the age of 4 years, through the examination of personal journals and medical records kept by the mother of the twins. Examination of these journals revealed several differences in development between the twins, some as early as 6 months of age. In the first year of life, the infants already differed in language development, social development, sleep patterns, and sensitivity to pain. This rare opportunity to view early autistic development gives direction to developmental theories of autism and clinically useful cues to early signs of autism. PMID- 16036470 TI - Disorders of visuo-spatial cognition. PMID- 16036471 TI - The dynamics of language-related brain images. PMID- 16036472 TI - The neuropsychology of human memory. PMID- 16036473 TI - Different windows on executive functions. PMID- 16036474 TI - Integrating numerical indices of structure and function to optimize diagnostic sensitivity and specificity in screening for glaucoma. PMID- 16036475 TI - Performance of community-based glaucoma screening using Frequency Doubling Technology and Heidelberg Retinal Tomography. AB - PURPOSE: To assess the performance of a community-based glaucoma screening algorithm in the general population. METHODS: A total of 659 individuals aged 50 90 years were screened for glaucoma. Presenting visual acuity, family history of glaucoma, FDT perimetry, and HRT tests were assessed. Additional samples of participants served as control groups. Participants identified as glaucoma positive received a full ophthalmic examination. Based on this exam a consensus diagnosis was made which served as the gold standard. RESULTS: The optimal screening strategy combining visual acuity and family history with FDT and HRT had sensitivities, specificities, positive predictive values and negative predictive values of 96.8%, 89.7%, 31.9%, and 99.8% respectively for detecting glaucoma. CONCLUSIONS: By combining assessments of presenting visual acuity and family history of glaucoma with Frequency Doubling Technology perimetry and Heidelberg Retina Tomography, we devised a community glaucoma-screening algorithm that showed a high sensitivity and specificity for detecting glaucoma in the general population. PMID- 16036476 TI - Vision and self-rated health: longitudinal findings from an older Australian population. AB - PURPOSE: To assess temporal associations between vision and self-rated health. METHODS: We examined 3654 Australian residents (82.4% response) aged 49+ years in the Blue Mountains Eye Study, during 1992 to 1994, and re-examined 2334 in 1997 to 1999. Change in vision was defined if baseline and 5-year visual acuity (VA) changed by > or =10 letters (2 lines). Change in self-rated health was defined if overall health ranking (excellent, good, fair, poor) changed by > or =1 step. RESULTS: Of 2285 participants with data from both examinations, 59.8%, 23.9%, and 16.3% reported unchanged, declined, and improved self-rated health, respectively. Among persons whose self-rated health declined, the proportions with a decline in presenting VA, no change, or improvement were similar (25.2%, 23.8% and 22.5%, respectively, p for trend = 0.5). Among participants with decline, no change, or improvement in self-rated health, a similar magnitude in the age-sex adjusted mean VA decline (-1.0 to -2.7 letters) was observed. After adjusting for age, sex, hospitalizations, and incident chronic conditions, declining VA was not associated with increased decline in 5-year self-rated health (multivariate adjusted odds ratio 0.99, 95% CI 0.75-1.30). CONCLUSION: There were no parallel 5 year changes in self-rated health and vision in this population. PMID- 16036477 TI - Ocular trauma in a Greek population: review of 899 cases resulting in hospitalization. AB - PURPOSE: To determine the annual hospitalization rate and risk factors of eye injuries requiring hospitalization in an area of Greece. METHODS: Retrospective case analysis of 899 consecutive patients with ocular injury admitted at the Department of Ophthalmology, University Hospital at Patras Medical School, Greece, over a 9-year period. RESULTS: The average annual rate of hospitalized ocular injuries was 71.0 eye injuries per 1000 admissions at the Department of Ophthalmology. The majority (80.2%) of patients were male. The average age was 35.0 years (range 6 months-98 years). Approximately half of all injuries occurred in persons younger than 29 years of age. Patients with Albanian ethnic background were at higher risk. The most common type of injury was closed-globe injury (49.5%) that affected more frequently the very young individuals (0-19 years) and persons older than 50 years of age. Open-globe injuries occurred most often in young adults, 20 to 39 years of age, and comprised over 35% of all eye injuries in this age group. Most injuries (32.8%) occurred at the workplace, most commonly during construction activities. Injuries at home approximated the work-related injuries (30.0%). A total of 398 (42.5%) eyes underwent surgical intervention on the eye globe, while 49 (12.3%) of them underwent multiple operations as part of their treatment. 18.2% of the eyes were blinded and 27.9% had significant visual acuity loss. An enucleation during the follow-up period was performed in 4 (1.0%) eyes. CONCLUSIONS: Trauma is an important cause of ocular morbidity in Greece. Risk factors include young age, male gender, Albanian ethnic background, and workplace. General safety precautions and behavior modification are indicated. PMID- 16036478 TI - A population-based case-control study of isolated ocular coloboma. AB - AIMS: To attempt the detection of possible etiological factors in the origin of isolated ocular, mainly iris coloboma. METHODS: The data set of the Hungarian Case-Control Surveillance of Congenital Abnormalities, 1980-2002, was used. Exposure data and family history were based on (1) prospective medical records, (2) retrospective maternal information, and (3) information obtained by regional nurses at home visit. RESULTS: Of 46 cases, 40 were affected with iris coloboma and 19 had bilateral manifestation. The positive family history indicated a hereditary origin in 4.3% (2/46) of cases. An association was found between the isolated ocular coloboma and hydroxyethylrutoside treatment (adjusted POR with 95% CI: 5.4, 2.2-12.9). Another association was seen between isolated ocular coloboma and hypothyroidism (adjusted OR with 95% CI = 12.6, 3.0-52.7), but it was based only on two cases. CONCLUSIONS: It is necessary to study the preventable environmental factors in the origin of these ocular defects. PMID- 16036479 TI - Vision and hearing impairment in aged care clients. AB - PURPOSE: To determine the frequency of sensory impairments among aged care clients. METHODS: We examined 188 persons aged 65-99 years being evaluated for aged care services at a geriatric assessment center, west of Sydney, Australia. Visual acuity was measured using a LogMAR chart. Visual impairment was defined in the better eye: mild < 20/40 to > or = 20/80, moderate < 20/80 to > or = 20/200, and severe < 20/200. Hearing function was assessed by portable pure-tone air conduction audiometry. Hearing loss was defined using average hearing thresholds in the better ear: mild > 25 to < or = 40 decibels (dB), moderate > 40 to < or = 60dB, and severe > 60dB. RESULTS: Vision, hearing, or both were randomly tested in 96, 93, and 49 aged care clients, respectively. Vision impairment was found in 30.2% of clients. Distance vision could be improved with pinhole in 16.7% of 84 clients with presenting VA < 20/20. Moderate to severe hearing loss was present in 50.5%. Combined sensory impairment was detected in 22.5% of persons having both tests. The age-standardized proportions with vision impairment was 25.6%, higher than the rate (17.4%) found in the Blue Mountains Eye Study (BMES) community population, but lower than the 37.7% prevalence reported from the BMES nursing home sample. Hearing impairment was also more common in aged care clients (28.1% vs 17.5%). CONCLUSIONS: This study suggests a high prevalence of sensory impairment among older persons transitioning from independent community living to institutionalized care. PMID- 16036480 TI - Determinants of myopia among Omani school children: a case-control study. AB - BACKGROUND: Our aim was to determine risk factors for myopia in students of 10th grade of eight regions of Oman. METHODS: This was a case control study. Students of 10th grade with and without myopia were cases and controls respectively. In 2002 to 2003, their vision, refractive status, and physical measurements were recorded. The health records of all students were reviewed for their ocular and physical status in the 7th grade. The association of myopia to anthropometric variables, "Protein Energy Malnourishment" status, gender, history of myopia in parents and sibling, trachoma, and allergic conjunctivitis were analyzed. RESULTS: Myopia in students was significantly associated to myopia in parents [OR = 2.11 (95% CI 1.8-2.47)] and their siblings [OR = 2.87 (95% CI 2.45-3.35)]. Height was significantly higher in myopes compared to non-myopes both at the 10th and 7th grades. Weight and body mass index were greater in myopes than non-myopes only at 7th grade. Myopia in parents and siblings and female gender were predictors of myopia in the students. CONCLUSIONS: Hereditary and anthropometric factors seem to be associated with myopia in Omani children. The genetic etiology needs further confirmation. PMID- 16036481 TI - Post-cycloplegia myopic shift in an older population. AB - PURPOSE: To demonstrate that use of a mydriatic agent remains a significant confounder in autorefraction of the presbyopic population. METHODS: The pre- and post-cycloplegic autorefraction results of 37 subjects over 50 years of age were measured using a Humphrey-598 autorefractor. The results of both eyes were included in a multivariate regression analysis. RESULTS: The average age of the patient sample was 63.4 years. The mean spherical equivalent (SEQ) shifts for the hyperopic, myopic, pseudophakic, and emmetropic eyes were -0.53 D (95% CI -0.77 D to -0.39 D; p < 0.001), -0.38 D (95%CI -0.81 D to +0.04 D; p = 0.077), -0.49 D (95% CI -0.78 D to -0.20 D; p = 0.001), and -0.35 D (95% CI -0.59 D to -0.11 D; p = 0.004), respectively. CONCLUSION: Cyclopleged autorefraction in the presbyopic population is associated with a myopic shift that can potentially lead to overestimation of myopic prevalence. This is an important factor in comparing population studies where cyclogleged autorefraction is used in contrast to non cyclopleged autorefraction and subjective refraction. PMID- 16036482 TI - Is smoking a causative factor of hypertension? PMID- 16036483 TI - Combination of antihypertensive drugs from a historical perspective. PMID- 16036484 TI - Acute and chronic effects of cigarette smoking on arterial stiffness. AB - OBJECTIVES: Pulse wave velocity (PWV) is an indicator of arterial stiffness, especially in the aorta, and a marker for vascular damage. Smoking is reported to increase arterial stiffness. We examined the acute and chronic effects of smoking on arterial stiffness by measuring brachial--ankle PWV (baPWV) using an oscillometric method (VP 1000, Colin Co., Komaki, Japan). METHODS: All healthy male subjects (chronic smokers, n=40, 30.3 years old vs non-smokers, n=40, 28.3 years old) smoked two cigarettes (nicotine 1.5 mg) within 10 min and measured blood pressure (BP), heart rate and baPWV at baseline, 5, 15, 30, 45 and 60 min and compared with controls (n=20, 29.3 years old). RESULTS: Systolic BP was higher in chronic smokers than non-smokers or controls. Smoking increased the systolic and diastolic BP and heart rate significantly at 5 min in both chronic smokers and non-smokers as compared with baseline levels or controls (respectively, p<0.001) and returned to baseline level at 15 min. Pulse pressure did not increase significantly. baPWV increased significantly in both chronic smokers and non-smokers at 5 min (12.1--17.3 m/s vs 11.1--12.7 m/sec, respectively) and remained higher for 30 min compared with controls (p<0.001). Smoking increased baPWV to a greater extent in chronic smokers than in non smokers (p<0.01). CONCLUSION: Acutely, cigarette smoking increased BP, heart rate and baPWV in chronic smokers and non-smokers. These effects were more prominent in chronic smokers than in non-smokers. These findings suggest that cigarette smoking have deleterious effects on cardiovascular system by stiffening arteries. PMID- 16036485 TI - Selected climatic variables and blood pressure in Central European patients with chronic renal failure on haemodialysis treatment. AB - BACKGROUND/AIMS;Higher blood pressure (BP) in winter has been documented in healthy and hypertensive adults. It may potentially contribute to the observed excess winter cardiovascular mortality in the general population. The aim of the study was to assess whether BP varies similarly among patients with chronic renal failure on haemodialysis treatment, who present an increased risk of cardiovascular death. METHODS: We retrospectively analysed values of pre-dialysis BP and parameters of fluid retention--pre-dialysis body weight and inter-dialytic weight gain measured in 49 patients (23 male, 26 female; aged 46.0+/-13.5 years) from 1995 to 1998. For each patient we calculated deviations of monthly mean values of systolic BP, diastolic BP, pre-dialysis body weight and inter-dialytic weight gain from the lowest monthly means of these parameters in a given year. Monthly means of these deviations for the whole study group (dSBP, dDBP, dBW, dWG, respectively) were subsequently computed. Monthly means of air temperature (T), air relative humidity (H) and atmospheric pressure (AP) were provided by the local Institute of Meteorology. The Wilcoxon paired test was applied to compare mean values of BPs and parameters of fluid retention of every patient in three warmest and three coldest months of each year. Spearman rank correlation analysis was employed to evaluate relationships between dSBP, dDBP and climatic variables, dBW or dWG. RESULTS: Systolic BP was higher in summer than in winter (146.6+/ 20.5 vs 143.4+/-18.9 mmHg; p<0.00001). Diastolic BP was also higher in summer than in winter (82.6+/-8.5 vs 79.6+/-7.3 mmHg; p<10(-9)). Pre-dialysis body weight and inter-dialytic weight gain did not differ between summer and winter (66.0+/-13.2 vs 66.0+/-13.2 kg; p=0.98 and 2.27+/-0.6 vs 2.29+/-0.5 kg; p=0.53). There was a positive correlation between dSBP and T (RS=0.424, p<0.003), as well as dDBP and T (RS=0.591, p<0.00001) and an inverse correlation between dSBP and H (RS=-0.372, p<0.01), as well as dDBP and H (RS=-0.408, p<0.004). There were no significant associations between BPs and AP, dBW or dWG. CONCLUSIONS: In haemodialysed patients from southern Poland, BP is higher in summer than in winter. Changes in BP are related to seasonal changes in climatic variables--air temperature and air relative humidity. Seasonal variation in BP is not associated with variation in fluid retention. Possible alteration of cardiovascular reactivity to changes in climatic environment in haemodialysed chronic renal failure patients may be one of the potential explanations of these observations. PMID- 16036486 TI - Tissue Doppler imaging of the left ventricle in healthy elderly females does not support the concept of "isolated" diastolic dysfunction. AB - The concept of "isolated" left ventricular diastolic dysfunction has recently been challenged, since left ventricular contraction abnormalities can be demonstrated in subjects with normal left ventricular ejection fraction (EF). In this study of 35 healthy females (aged 65--80 years) with EF>50%, the ratio of peak early transmitral flow velocity to peak early diastolic myocardial velocity (E/Em) correlated significantly with peak systolic myocardial longitudinal velocity (Sm) (r=-0.57, p<0.0001), assessed as an average in six basal left ventricular segments. E/Em correlated also with age (r=0.51, p<0.002), but not significantly with ambulatory daytime systolic blood pressure (r=0.32, p=0.06), nor with left ventricular mass. In conclusion, there is a significant correlation between left ventricular diastolic and systolic function also in an apparently healthy population when adequately sensitive methods are used, in terms of tissue Doppler assessment of the left ventricular longitudinal motion. Although age may influence left ventricular longitudinal motion, an influence of arterial blood pressures is unclear. PMID- 16036487 TI - Plasma NT-proBNP concentration is related to ambulatory pulse pressure in peripheral arterial disease. AB - BACKGROUND: Increased levels of N-terminal prohormone brain natriuretic peptide (NT-proBNP) are associated with left ventricular dysfunction (LVD) and left ventricular hypertrophy (LVH), but the relation of NT-proBNP to ambulatory blood pressure (ABP) and hypertensive target organ damage in high-risk patients with peripheral arterial disease (PAD) has not been studied. We hypothesized that NT proBNP levels were increased in patients with PAD in comparison to a matched control group and that levels of NT-proBNP were related to ABP. METHODS: Blood samples were analysed for NT-proBNP in 103 males with PAD and 96 age- and sex matched controls. Ninety-eight PAD patients performed ABP monitoring and 99 underwent Tc-99m Sestamibi myocardial perfusion SPECT. RESULTS: NT-proBNP was significantly increased in PAD patients compared with controls [median (interquartiles)] 167(76, 418) vs 68(38, 142) pg/ml, p<0.001. Plasma levels of NT proBNP correlated positively to systolic blood pressure (SBP), pulse pressure (PP), night-day ratio of SBP and showed the strongest correlation to average night PP (r=0.42, p<0.001). In multiple regression analysis, night PP remained independently related to NT-proBNP. CONCLUSION: NT-proBNP levels are markedly increased in PAD patients compared to age-matched controls. Night PP is related to NT-proBNP levels independently of other variables highlighting the importance of ambulatory PP as a cardiovascular risk factor. Measurement of NT-proBNP could be indicated in PAD patients for further risk stratification. PMID- 16036488 TI - Cost analysis of different pharmacological treatment strategies in elderly hypertensives. AB - OBJECTIVE: To compare costs for management of hypertension in elderly hypertensives randomized to starting treatment with conventional (beta blockers/diuretics) therapy or a therapy initiated with a calcium antagonist or an angiotensin-converting enzyme (ACE) inhibitor. DESIGN: Health economic substudy in the Swedish Trial in Old Patients with Hypertension-2 (STOP Hypertension-2). SETTING: Outpatient clinics in Sweden. In this health economics substudy, 16/312 participating STOP-2 trial centers were selected. SUBJECTS: Elderly (70--84 years) patients (n=303) with a systolic and/or diastolic hypertension (or=180 and/or 105 mmHg). METHODS: Costs for patient management were analyzed and categorized in costs for routine care (protocol-driven costs, PDC), costs for extra visits or care (non-protocol-driven costs, NPDC), and direct drug costs (drug treatment costs, DTC). All calculations are related to costs during the first year of treatment after inclusion in STOP Hypertension-2. RESULTS: Out of the scheduled visits, a total of 99% were actually performed by the patients. There were no differences in the number of visits between the three treatment groups (diuretics/beta-blockers, calcium antagonists or ACE inhibitors). PDC did thus not differ between the three treatment groups. NPDC were similar in the conventional and calcium antagonist groups and lower than for the ACE inhibitor group. DTC were lower in the conventional treatment group compared with the other two groups. CONCLUSION. In elderly hypertensives in STOP Hypertension-2, total costs for management of hypertension were lower in patients assigned to diuretics, beta-blockers or calcium antagonists compared with ACE inhibitors during the first year of treatment. These results may be relevant to management of elderly hypertensive patients, especially in those patients without compelling indications or contraindications to starting treatment with either of these three main drug alternatives. Notably, with a specific drug regimen there are sizable NPDC such as extra visits and controls associated with symptoms or side-effects of a specific therapy, which significantly add to the total costs of treatment. Such costs, beyond the actual costs for the drugs, are important to realize and evaluate in order to provide the true costs for treatment of hypertensive patients. PMID- 16036489 TI - Endothelin A antagonist LU-135252 and trandolapril in the treatment of the Cohen Rosenthal diabetic hypertensive rat. AB - BACKGROUND: Hypertension and non-insulin-dependent diabetes mellitus (NIDDM) often occur simultaneously and the combination requires vigorous control of hypertension. This can generally be achieved by a combination of antihypertensive drugs. The present study examines the antihypertensive and possible hypoglycemic effects of combined therapy with endothelin A (ETA) receptor antagonist LU-135252 and angiotensin-converting enzyme (ACE) inhibitor trandolapril in male Cohen Rosenthal Diabetic Hypertensive (CRDH) rats. METHODS: Rats were divided into four groups as follows: group I served as control; group II--LU-135252 30 mg/kg/day; group III--trandolapril 0.1 mg/kg/day and group IV--both LU-135252 30 mg/kg/day and trandolapril 0.1 mg/kg/day. Systolic blood pressure (SBP) and plasma glucose levels were evaluated at the beginning of the experiment and after 2, 4 and 6 weeks. RESULTS: SBP decreased significantly in all treated groups after 2, 4 and 6 weeks of treatment compared to baseline. Maximum decrease was in group IV (combination) from 174.8+/-3.7 to 136.1+/-2.4 mmHg (22%) (p<0.0001); in group III (trandolapril) from 165.8+/-2.7 to 137.5+/-2.9 mmHg (17%) (p=0.0002); and in group II (LU-135252) and from 169.1+/-3.1 to 147.8+/-2.5 mmHg (12%) (p=0.0004). Glucose levels in plasma decreased significantly after 6 weeks of treatment. Maximum decrease was in group IV: from 501.0+/-42.8 to 178.6+/-7.3 mg/dl (62%) (p<0.0001); in group III from 428.2+/-47.7 to 146.8+/-5.6 mg/dl (63%) (p<0.0001); and in group II from 491.2+/-39.3 to 272.2+/-28.3 mg/dl (42%) (p=0.0002). CONCLUSION: The SBP decrease was additive when both drugs were given together. Thus, combination of ETA antagonist and ACE inhibitor appears to offer a rational fixed-dose antihypertensive therapy, which is superior to that of either drug alone. The decrease in glucose level, which was the least impressive while on LU 135252 alone, was more prominent during combination after 2 weeks, although without further decrease. PMID- 16036491 TI - Health economy of the metabolic syndrome pandemic. PMID- 16036490 TI - Effects of allopurinol, a xanthine oxidase inhibitor, on renal injury in hypercholesterolemia-induced hypertensive rats. AB - To investigate if increased lipid peroxidation is involved in hypercholesterolemia-induced hypertension and renal injury, we examined the effects of allopurinol, a xanthine oxidase inhibitor, on these conditions. Groups of male Sprague--Dawley rats were fed for 8 weeks with a high-cholesterol diet (4% cholesterol), a high-cholesterol plus allopurinol (10 mg/kgBW/day) diet or a normal diet. Systolic blood pressure (SBP), serum lipids, uric acid (UA) and malondialdehyde (MDA) as a measure of lipid peroxides, and urinary excretion of protein (UP) were measured after 0, 4 and 8 weeks. Urinary excretion of nitrite plus nitrate (UNOx) and iron (UFe), and MDA in the kidney were measured after 8 weeks. The renal injury was evaluated by the glomerular sclerosis score (SS). The high-cholesterol diet increased SBP, serum total cholesterol and UA, MDA in the serum and kidney, UP, UNOx, UFe and SS. Allopurinol ameliorated cholesterol induced elevation in serum UA, MDA in the serum and kidney, UP, UNOx, UFe and SS, but did not affect SBP. Hence, our results suggest that lipid peroxidation may be involved in hypercholesterolemia-induced renal injury, and that suppression of lipid peroxidation can reduce such injury. PMID- 16036492 TI - Who's really hypertensive?--Quality control issues in the assessment of blood pressure for randomized trials. AB - The characterization of blood pressure in treatment trials assessing the benefits of blood pressure lowering regimens is a critical factor for the appropriate interpretation of study results. With numerous operators involved in the measurement of blood pressure in many thousands of patients being screened for entry into clinical trials, it is essential that operators follow pre-defined measurement protocols involving multiple measurements and standardized techniques. Blood pressure measurement protocols have been developed by international societies and emphasize the importance of appropriate choice of cuff size, identification of Korotkoff sounds, and digit preference. Training of operators and auditing of blood pressure measurement may assist in reducing the operator-related errors in measurement. This paper describes the quality control activities adopted for the screening stage of the 2nd Australian National Blood Pressure Study (ANBP2). ANBP2 is cardiovascular outcome trial of the treatment of hypertension in the elderly that was conducted entirely in general practices in Australia. A total of 54 288 subjects were screened; 3688 previously untreated subjects were identified as having blood pressure >140/90 mmHg at the initial screening visit, 898 (24%) were not eligible for study entry after two further visits due to the elevated reading not being sustained. For both systolic and diastolic blood pressure recording, observed digit preference fell within 7 percentage points of the expected frequency. Protocol adherence, in terms of the required minimum blood pressure difference between the last two successive recordings, was 99.8%. These data suggest that adherence to blood pressure recording protocols and elimination of digit preferences can be achieved through appropriate training programs and quality control activities in large multi centre community-based trials in general practice. Repeated blood pressure measurement prior to initial diagnosis and study entry is essential to appropriately characterize hypertension in these elderly patients. PMID- 16036493 TI - First morning standing up may be risky in acutely ill older inpatients. AB - Information about orthostatic hypotension (OH) among elderly patients hospitalized for acute conditions in short-term facilities is scarce. Many older inpatients carry various predisposing factors for OH. However, its existence goes frequently unrecognized. In this context, first morning standing up following admission for an acute disease may be dangerous. The aim of this study was to investigate OH and associated manifestations in this situation. OH (> or = 20 mmHg systolic and/or (> or =10 mmHg diastolic blood pressure fall), heart rate, arrhythmias and appearance of dizziness or palpitations were recorded in 121 sequential inpatients aged >65 years, prior to and 1, 3 and 5 min following first morning standing. OH occurred in 64.5% of patients, while dizziness and/or palpitations appeared in 76%. Severe adverse effects were registered in 11.5% of OH patients. Significantly associated with OH were: bed rest lasting 9-24 h (vs (< or = 8 h, p<0.001), appearance of dizziness or palpitations (p<0.001 and p=0.005, respectively), heart failure (p=0.02) and renal dysfunction (p=0.04). OH and/or associated symptoms are frequent in acutely ill older inpatients on first morning standing up following nocturnal bed rest. The ominous potential consequences call for alertness to this phenomenon and application of appropriate preventive measures. PMID- 16036494 TI - Hypertensive smokers have a worse cardiovascular risk profile than non-smokers in spite of treatment--a national study in Sweden. AB - Smoking is a well-established risk factor for cardiovascular disease. Studies have indicated that smoking may outweigh the benefit of blood pressure (BP) control. Our aim was to compare cardiovascular risk factors in smokers vs non smokers from a national sample of treated hypertensives. Data were collected on smoking habits, BP control, total and low-density lipoprotein (LDL) cholesterol, diabetes, left ventricular hypertrophy (LVH), and microalbuminuria (MA), from records of 4424 consecutive patients by 189 physicians. All technical methods were local. Treated hypertensives who smoked had microalbuminuria significantly more often than non-smokers, 26.2% vs 20.5% (p<0.05), and a higher proportion of smokers were suboptimally controlled (DBP > or = 90 mmHg), 32.7% vs 25.0% (p<0.01). Smoking males had a higher prevalence of LVH (25.7% vs 20.1; p<0.05), microalbuminuria (29.7% vs 24.7%; p<0.01), and a higher proportion of subjects with uncontrolled systolic BP (> or = 140 mmHg) (72.8% vs 68.9%; p<0.01). Both DBP and total cholesterol were higher in smoking vs non-smoking females. An increased prevalence of LVH and microalbuminuria was independently associated with smoking. In summary, smokers with treated hypertension show a higher proportion of LVH (men), microalbuminuria and worse diastolic BP control than non smokers. This may hypothetically reflect either less compliance with drug treatment in smokers or that smoking impairs the pharmacological effects of antihypertensive drugs. PMID- 16036495 TI - Efficacy of telephone and mail intervention in patient compliance with antihypertensive drugs in hypertension. ETECUM-HTA study. AB - OBJECTIVE: To study the efficacy of telephone and mail intervention in therapeutic compliance among patients with mild to moderate hypertension. DESIGN: A prospective controlled multicenter clinical trial. SETTING: Eighty-five primary care centers in Spain, with a duration of 6 months. PATIENTS: A total of 636 patients with newly diagnosed or uncontrolled hypertension were included. Interventions. The patients were randomized and distributed between the following groups: (i) control (CG) - under routine clinical management; (ii) mail intervention (MIG) - received a mailed message reinforcing compliance and reminding of the visits (15 days, 2 and 4 months); (iii) telephone intervention (TIG) - received a telephone call at 15 days, then at 7 and 15 weeks. MAIN OUTCOME MEASURE: Five visits were scheduled, with the measurement of blood pressure and counting of tablets. Compliers were defined as subjects showing 80 110% drug consumption. Calculations were made of mean percentage compliance (MPC) and compliers, mean blood pressure and percentage controlled subjects. RESULTS: Five hundred and thirty-eight patients completed the study (261 males); 85.5% were compliers (CI = 82.5-88.5; n = 460). The MPC was 95.1+/-19.6% (CI = 93.28 96.92). The CG consisted of 182 individuals, MIG = 172 and TIG = 184. Compliers represented 69.2% of the CG (CI 62.5-75.9%), 91.3% (CI = 87.1-95.5) of the MIG (p = 0.0001) and 96.2% of the TIG (CI 93.5-98.9%); the final MPC was 89.6%+/-15 in CG, 96.6%+/-12 in MIG and 99.1+/-26.8 in TIG (p = 0.0001). The percentage of controlled subjects was 47.2% in CG (CI = 40-54.4), 61.3% in MIG (CI = 54.1 68.5%) and 63.3% in TIG (CI = 56.4-70.2%) (p<0.05). CONCLUSIONS: TIG and MIG are effective measures for improving patient compliance in hypertension. PMID- 16036496 TI - Blood viscosity: effects of mental stress and relations to autonomic nervous system function and insulin sensitivity. AB - We studied effects of mental stress on whole-blood viscosity (WBV) and blood pressure (BP), and relations between WBV and autonomic nervous system activity and insulin sensitivity. We measured WBV (rotational rheometer), plasma noradrenaline (NA), finger BP, heart rate variability (HRV) and baroreflex sensitivity (BRS; transfer technique) during hyperinsulinaemic glucose clamp and mental arithmetic stress test (MST) in 20 men with high ( > or =140/90 mmHg) and 21 men with normal (< or =115/75 mmHg) screening BP, and 10 women regardless of screening BP (all normotensive). WBV and NA increased during the MST, while HRV and BRS decreased. During the MST, WBV (all shear rates) and the response ((delta)WBV) (low shear) were higher in men with high compared to normal screening BP (p<0.05). In men, WBV correlated positively with NA and negatively with HRV, BRS and insulin sensitivity. The diastolic BP response ((delta)DBP) was independently explained by high-shear (delta)WBV (p<0.05) and (delta)NA (p<0.0001), and (delta)WBV independently by (delta)DBP (p<0.05). WBV is related to increased sympathetic activity, impaired vagal cardiac control and low insulin sensitivity in young adults. The haemorheological effect of mental stress is increased in young men with high screening BP and may be mediated by the acute increase in BP. PMID- 16036497 TI - Controlling systolic blood pressure is difficult in patients with diabetic kidney disease exhibiting moderate-to-severe reductions in renal function. AB - This study compared the use of antihypertensive treatment and blood pressure (BP) controls between patients with diabetic kidney disease (DK+) and patients with non-diabetic kidney disease (DK-) exhibiting moderate-to-severe chronic renal failure who did not need renal replacement therapy. A cross-sectional survey included all renal patients with s-creatinine at ?200 micromol/l attending regular control sessions at six renal units in Norway. Of the 351 patients included, 73 (20.8%) were DK+. The proportion reaching a BP goal of <130/80 mmHg was similar in DK+ and DK- (14.1% vs 13.6%, p = 0.92), while 38% and 39% achieved a BP of <140/90 mmHg, respectively. The systolic BP goal was more difficult to achieve than the diastolic BP goal in DK+ patients (35% vs 15%) despite a mean of three different types of drugs being used. Loop diuretics and beta-adrenergic receptor antagonists were the most frequently prescribed drugs, and the use of angiotensin-converting enzyme inhibitors or angiotensin-II-receptor antagonists declined when renal function deteriorated, from 80% to 0% and from 66% to 20% in the DK+ and DK- groups, respectively (p = 0.001). Thus, despite the use of multiple antihypertensive drugs, controlling BP - especially the systolic BP - is difficult in high-risk patients with chronic renal failure caused by diabetic kidney disease. PMID- 16036498 TI - Losartan but not atenolol reduce carotid artery hypertrophy in essential hypertension. A LIFE substudy. AB - BACKGROUND: We wanted to investigate whether treatment with losartan, an angiotensin II receptor blocker, induced regression of carotid artery hypertrophy as compared to the beta-receptor blocker, atenolol. METHODS: In 45 patients recruited for the LIFE Study with stage II-III hypertension and ECG left ventricular (LV) hypertrophy, we measured blood pressure, intima-media thickness (IMT) and lumen in the common carotid arteries by ultrasound, and minimal forearm vascular resistance (MFVR) by plethysmography, after 2 weeks of placebo treatment and after 1, 2 and 3 years of anti-hypertensive treatment with either atenolol- or losartan-based regimens. We measured the same parameters in 26 normal subjects matched for age and gender. RESULTS: The patients had as compared to normotensive controls higher IMT (0.87 vs 0.76 mm, p = 0.001) and intima-media cross-sectional area (IMA) (19.7 vs 15.5 mm2, p<0.001). Systolic and diastolic blood pressures were reduced to the same degree in patients treated with losartan as compared to atenolol. However, IMA decreased significantly only in patients treated with losartan (19.2 vs 17.6 mm2, p = 0.001) and the average relative decrease in IMA during the 3 years of treatment was significantly higher in patients treated with losartan as compared to atenolol (-7.4 vs -2.0%, p<0.05). CONCLUSION: Patients with hypertension and LV hypertrophy had hypertrophy of the common carotid arteries. Losartan, but not atenolol, induced regression of this hypertrophy. Because carotid artery hypertrophy has been associated with strokes, our findings may explain the lower incidence of strokes in the LIFE study in patients treated with losartan as compared to atenolol. PMID- 16036499 TI - Effects of olmesartan and enalapril at low or high doses on cardiac, renal and vascular interstitial matrix in spontaneously hypertensive rats. AB - We have evaluated the effects of different doses of an angiotensin-converting enzyme (ACE) inhibitor, enalapril (ENA) and of an angiotensin II type 1 receptor blocker olmesartan (OLM), on extracellular matrix of the heart, kidney, aorta and mesenteric artery of spontaneously hypertensive rats (SHR). Forty SHR and eight Wistar-Kyoto controls (WKY) were included in the study. Eight SHR were treated with high-dose OLM 15 mg/kg per day, eight with high-dose ENA 25 mg/kg per day, eight with low-dose OLM 1 mg/kg per day and eight with low-dose ENA (2 mg/kg per day). Eight SHR and eight WKY were kept untreated as controls. Treatment was from age 4 to 12 weeks. Systolic blood pressure (SBP) was measured non-invasively every week. The left ventricular weight to body weight (RLVM) was measured, and the cardiac, aortic and glomerular interstitial collagen content was evaluated using Sirius red staining and image analysis. Mesenteric small arteries were dissected and mounted on a micromyograph, and the media:lumen ratio (M/L) was calculated. Collagen subtypes were evaluated by polarized light microscopy. The SHR treated with high-dose OLM or ENA showed a normalization of SBP. The RLVM was significantly increased in untreated SHR compared with untreated WKY, whereas significantly lower values were observed in the groups of SHR treated with high dose OLM or ENA. A significant increase in cardiac and glomerular collagen content was observed in untreated SHR. Both high- or low-dose OLM and ENA normalized collagen content in the heart and the kidney. Both high-dose OLM and high-dose ENA normalized M/L ratio; however, OLM proved to be more effective than ENA in normalizing collagen pattern. In fact, aortic collagen content was normalized by both high-dose and low-dose OLM, but only by high-dose ENA. In conclusion, both OLM and ENA were significantly and equally effective in the prevention of cardiac and renal damage in SHR, whereas OLM was more effective than ENA in terms of effects on vascular extracellular matrix. PMID- 16036500 TI - Pathophysiological basis of pharmacotherapy in the hepatorenal syndrome. AB - Hepatorenal syndrome (HRS) is a functional and reversible impairment of renal function in patients with severe cirrhosis. Major pathophysiological elements include liver dysfunction, a circulatory derangement with central hypovolaemia and neurohumoral activation of potent vasoactive systems leading to a pronounced renal vasoconstriction. The prognosis of patients with HRS is poor but recent research has spread new enthusiasm for treatment. Efforts at treatment should seek to improve liver function, to ameliorate arterial hypotension and central hypovolaemia, and to reduce renal vasoconstriction. Therefore a combined approach should be applied with reduction of portal pressure with e.g. ss-adrenergic blockers and transjugular intrahepatic portosystemic shunt (TIPS), with amelioration of arterial hypotension and central hypovolaemia with vasoconstrictors such as terlipressin and plasma expanders. New experimental treatments with endothelin- and adenosine antagonists and long-acting vasoconstrictors may have a future role in the management of HRS. PMID- 16036501 TI - Oesophageal defensin expression during Candida infection and reflux disease. AB - OBJECTIVE: Alpha (alpha-) and beta (beta-) defensins are major constituents of the innate defence system, providing rapid antimicrobial action. The expression of alpha- and beta-defensins in the oesophagus and stomach by quantitative, real time, polymerase chain reaction (PCR) in healthy individuals was studied to define the influence of oesophageal Candida infection on their expression in comparison to oesophageal reflux disease. MATERIAL AND METHODS: Biopsy samples were taken from the upper gastrointestinal tract, mRNA was extracted, reverse transcribed into cDNA and real-time reverse transcription PCR (RT-PCR) analysis measuring transcript number of a-defensins and ss-defensins performed. Standard curves allowed quantification of gene copies per weight of mRNA. RESULTS: hBD-1, hBD-2 and hBD-3 had their highest expression levels in the oesophagus with factor 3 to 5 lower in the stomach. Candida oesophagitis resulted in massive up regulation of hBD-2 (800-fold), while hBD-1 and hBD-3 expression were slightly increased. In addition, expression of HNP 1-3 was detected, indicating infiltration of neutrophil granulocytes. In reflux disease, an up-regulation of hBD-2 (20-fold) and hBD-3 (50-fold) could also be observed, while hBD-1, hBD-4 and HD5 remained unaffected. Cytokine expression of interleukin-1beta, interleukin-6 and interleukin-8 were increased in both groups, while interleukin 10 expression was elevated only in reflux lesions. CONCLUSIONS: Candida colonization induced a high expression of antimicrobial peptides. In oesophageal reflux disease, induction of defensin expression could also be observed but to a lower degree. Therefore, up-regulation of defensins might protect against invasive candidiasis and keep the Candida infection limited to the mucosal surface. PMID- 16036502 TI - Gastroduodenal cytomegalovirus infection is common in kidney transplantation patients. AB - OBJECTIVE: Cytomegalovirus (CMV) infection is known to cause ulcerations, erosion and mucosal haemorrhage in the gastrointestinal tract. The aim of this study was to report the CMV findings in the gastroduodenal mucosa of kidney transplantation patients and immunocompetent controls. MATERIAL AND METHODS: Forty-six kidney transplant patients with upper gastrointestinal symptoms and 43 immunocompetent, dyspeptic patients (controls) prospectively underwent oesophagogastroduodenoscopies (OEGDs), with biopsies from the duodenum and stomach. CMV was demonstrated by immunohistochemistry, both in frozen sections using a monoclonal antibody against CMV-specific antigens (pp65 matrix protein) and in paraffin sections by means of a monoclonal antibody against the delayed early protein (p52). RESULTS: CMV was detected in the gastric mucosa in 30% of the kidney transplant patients and in 9% of the controls (p<0.05) and in the duodenal mucosa in 70% and 35%, respectively (p<0.01). The total frequency of CMV findings was similar in patients who underwent OEGDs <1 year and >1 year after transplantation. CMV inclusions were found only in transplantation patients <1 year after transplantation (n=9). CMV findings, especially inclusions, in the gastric biopsies were associated with nausea and upper gastric pain. Histopathological findings in CMV-positive samples were non-specific, focal inflammation in haematoxylin-eosin-stained preparations, while CMV p52 staining showed inclusions in either the epithelial or endothelial cells. CONCLUSIONS: CMV could be detected in the gastroduodenal mucosa in 74% of kidney transplantation patients and in 40% of immunocompetent controls (p<0.01). CMV diagnostics are always recommended when gastroduodenal biopsies of kidney transplantation patients are performed. PMID- 16036503 TI - Somatostatin inhibits gastric acid secretion more effectively than pantoprazole in patients with peptic ulcer bleeding: a prospective, randomized, placebo controlled trial. AB - OBJECTIVE: Gastric acid inhibition is beneficial in the management of peptic ulcer bleeding (PUB). The aim of this double-blind study was to test whether somatostatin (SST) increases intragastric pH in PUB as compared with pantoprazole (PAN) and placebo (PLA). MATERIAL AND METHODS: Eligible patients were randomized to receive SST (500 microg/h+250 microg bolus), or PAN (8 mg/h+80 mg bolus) or PLA (normal saline) i.v., for 24 h. All patients underwent gastric pH monitoring during the infusion of the trial drugs. RESULTS: The three groups (SST, n=14; PAN, n=14; PLA, n=15) were comparable for age, gender, aetiology of PUB and laboratory data at admission. Mean (+/-SE) baseline pH levels in the fundus increased during the administration of the trial drugs (SST: 1.94+/-0.18 to 6.13+/-0.37, p<0.0001; PAN: 1.93+/-0.16 to 5.65+/-0.37, p<0.0001; PLA: 1.86+/ 0.12 to 2.10+/-0.15, p=0.0917). During the first 12 h of infusion, the mean (+/ SE) percentage time spent above pH 4.0 and 5.4 was higher with SST versus PAN (84.4%+/-4.8 versus 55.1%+/-8.3, p=0.0049 and 74.2%+/-6.5 versus 47.1%+/-8.3, p=0.0163, respectively) and there was a trend favouring the SST group regarding the time spent above pH 6.0 and 6.8 (65.7%+/-6.4 versus 43.3%+/-8.2, p=0.0669 and 49.2%+/-7.7 versus 28.4+/-6.6, p=0.0738, respectively). CONCLUSIONS: In PUB, both SST and PAN inhibit gastric acid secretion as compared with placebo. However, during the first 12 h of the infusion, SST was more effective than PAN in maintaining high intragastric pH. These results may provide a rationale for the administration of SST in PUB. PMID- 16036504 TI - Evidence-based examination of the African enigma in relation to Helicobacter pylori infection. AB - OBJECTIVE: The African enigma describes the dissociation between the prevalence of Helicobacter pylori infection and H. pylori-related diseases. The aim of this study was to use an evidence-based review of endoscopic data from African countries to test whether there are data to support the concept of an African enigma. MATERIAL AND METHODS: A Medline search was carried out to identify prospective endoscopic studies in African populations. Data collected included: the number of endoscopies, age range (or mean age if available), indications for endoscopy, country, years during which data were collected, male to female ratio, and specific outcome of duodenal ulcer, gastric ulcer, or gastric cancer. RESULTS: Forty prospective endoscopic studies from 17 African countries were identified (20,531 patients) and evaluated between 1972 and 2001. Mean ages ranged from 31 to 53.1 years and male to female ratios from 0.67:1 to 4.64:1. H. pylori-related clinical outcomes were common; duodenal ulcers in 4326 patients (21.1%), gastric ulcers in 691 patients (3.4%), and gastric cancers in 503 patients (2.4%). CONCLUSIONS: Prospective upper endoscopic trials suggest that the clinical outcomes associated with H. pylori infection in Africa are similar to those seen in industrialized countries. No dissociation between the prevalence of H. pylori infection and H. pylori-related diseases existed; the African enigma as such does not exist and the continued study of the mechanism of a non-existent phenomenon is a misuse of resources. The myth resulted from reliance on anecdotal data and selection bias in populations with extremely limited access to health care and a relatively short life expectancy. PMID- 16036505 TI - Gastric mucosal cytokine levels in relation to host interleukin-1 polymorphisms and Helicobacter pyloricagA genotype. AB - OBJECTIVE: The outcome of a Helicobacter pylori infection is related in part to interrelationships among H. pylori virulence factors and the H. pylori-induced mucosal response. The host inflammatory response is partly governed by polymorphisms in pro-inflammatory genes. MATERIAL AND METHODS: Cytokine levels (interleukin (IL)-1beta, IL-6 and IL-8) were examined in H. pylori-infected and uninfected normal-appearing mucosa from patients with non-ulcer dyspepsia (NUD), margins of gastric ulcers and cancer tissues. Cytokine levels were compared with cagA genotypes and host interleukin (IL)-1 polymorphisms. RESULTS: The study comprised 168 Thai patients. All infected patients possessed anti-CagA antibody. Gastric mucosal IL-8 levels were significantly higher in H.pylori-positive cases than in -negative cases in all three tissue types (e.g. 1115 versus 217 pg/mg protein for NUD) (p<0.001). Normal-appearing but H. pylori-infected antral mucosa of patients with cagA type 1a strains had higher IL-8 levels than those with type 2a strains (2632 versus 1036 pg/mg protein) (p<0.005). IL-1B-511T/T carriers had higher antral mucosal IL-1ss levels versus non-carriers (pg/mg protein) (T/T=221, T/C=178, C/C=70) (p=0.005). IL-1B-511T/T carriers also had higher IL-1beta levels versus non-carriers in H. pylori-negative patients. CONCLUSIONS: It was found that both the host factors (IL-1 polymorphisms) and bacterial factors (cagA type 1a versus type 2a) influenced gastric mucosal cytokine levels. Future studies should concentrate on interactions among host factors (e.g. genetics and tissue responses) and bacterial and environmental factors. PMID- 16036506 TI - Development and validation of the Patient Assessment of Constipation Quality of Life questionnaire. AB - OBJECTIVE: Chronic constipation is characterized by difficult, infrequent, or seemingly incomplete bowel movements. The Patient Assessment of Constipation Quality of Life (PAC-QOL) questionnaire was developed to address the need for a standardized, patient-reported outcomes measure to evaluate constipation over time. MATERIAL AND METHODS: Items for the PAC-QOL were generated from the literature, clinical experts, and patients. Following principal components and multi-trait analyses, 28 items were retained forming four subscales (worries and concerns, physical discomfort, psychosocial discomfort, and satisfaction) and an overall scale. Validation studies were conducted in the United States, Europe, Canada, and Australia, to evaluate the internal consistency reliability (Cronbach's alpha), reproducibility (Intraclass Correlation Coefficients (ICCs)), validity (analysis of variance models), and responsiveness (effect size) of the PAC-QOL scales. RESULTS: The PAC-QOL scales were internally consistent (Cronbach's alpha >0.80) and reproducible (ICCs >0.70, except for the satisfaction subscale ICC=0.66). PAC-QOL scale scores were significantly associated with abdominal pain (p<0.001) and constipation severity (p<0.05). Effect sizes in patients reporting improvements in constipation over a 6-week period were moderate to large, with subscale effect sizes ranging from 0.76 to 3.41 and the overall scale effect size=1.77. Similar findings were observed in validation studies conducted in Europe, Canada, and Australia. CONCLUSIONS: The PAC-QOL is a brief but comprehensive assessment of the burden of constipation on patients' everyday functioning and well-being. Multinational studies demonstrate that the PAC-QOL is internally consistent, reproducible, valid, and responsive to improvements over time. PMID- 16036507 TI - Costs of outpatients with fecal incontinence. AB - OBJECTIVE: Fecal incontinence is a problem with a high prevalence. Patients generally suffer from their problems for many years. It has been shown that quality of life is negatively affected but health economic data for fecal incontinence are limited. The aim of this study was to estimate the costs associated with fecal incontinence in a large outpatient study group, taking a societal perspective. MATERIAL AND METHODS: Based on questionnaire data, we calculated the costs of health-care resources, out-of-pocket expenses and costs associated with production losses in paid and unpaid work. RESULTS: Data were available for 253 patients, of which 228 (90%) were female and 209 (83%) were treated in an academic medical center. The mean age of patients was 59 years (SD+/-13) with a mean duration of incontinence of 8.5 years (SD+/-8.3). Total costs were estimated on C2169 per fecal incontinent patient per year. Production losses in paid and unpaid work accounted for more than half of the total costs and costs of health-care visits accounted for almost a fifth of total costs. Costs associated with protective material (partially reimbursable and not reimbursable) formed only one-tenth of total costs, while incontinence medication was responsible for only 5% of total costs. CONCLUSIONS: More than half of total costs of fecal incontinence are made up of indirect non-medical costs. The costs associated with the use of incontinence material and other personal expenses are limited. PMID- 16036508 TI - Melatonin as a modulator of the ileal brake mechanism. AB - OBJECTIVE: The gastrointestinal tract represents the most important extrapineal source of melatonin. Intestinal melatonin release is induced by the ileal passage of nutrients and could play a part in the control of postprandial gut motility. The specific aim of this study was to determine the putative role of melatonin in the "ileal brake" reflex, an important mechanism released by ileal lipids that regulates the gastric emptying of chyme. MATERIAL AND METHODS: Under general anaesthesia rats were fitted with ileal cannula exteriorized at the back of the neck. After a 1-week recovery, experiments were performed in conscious fasted animals. Rats were fed by gavage 1.5 ml casein hydrolyse plus 0.05% phenol red and either saline or Intralipid were continuously infused (2 ml/h) into the ileum. Gastric emptying was measured 50 min after ingestion by gastric lavage and determination of phenol red by spectrophotometry. The effects of melatonin (1 mg/kg) and melatonin antagonist S-22153 (dose-response study 0.2-25 mg/kg) were tested versus vehicle in paired experiments at 1-week intervals. RESULTS: Ileal infusion of lipids delayed gastric emptying. During ileal infusion of lipids, melatonin antagonist S-22153, but not melatonin, potentiated the delay in gastric emptying induced by the ileal brake mechanism. The inhibition of gastric emptying induced by S-22153 was dose related. Neither melatonin nor S-22153 had noticeable effects on gastric emptying during ileal infusion of saline. CONCLUSIONS: Our data suggest that melatonin, released in response to ileal lipids, exerts a modulatory influence that decreases the inhibitory effects of the ileal brake on gastric emptying of nutrients. PMID- 16036509 TI - Small-bowel mucosal transglutaminase 2-specific IgA deposits in coeliac disease without villous atrophy: a prospective and randomized clinical study. AB - OBJECTIVE: In coeliac disease, autoantibodies directed against transglutaminase 2 are produced in small-bowel mucosa, and they have been found to be deposited extracellularly. The aim of this study was to investigate whether such mucosal IgA deposits are important in the diagnostic work-up of early-stage coeliac disease without small-bowel mucosal villous atrophy. MATERIAL AND METHODS: Forty one adults suspected of coeliac disease owing to increased density of mucosal gamma(delta)+ intraepithelial lymphocytes but normal villous morphology were randomized to gluten challenge or a gluten-free diet for 6 months. Clinically and histologically verified gluten dependency was compared with existence of small bowel mucosal transglutaminase 2-specific extracellular IgA deposits and (coeliac disease-type) HLA DQ2 and DQ8; 34 non-coeliac subjects and 18 patients with classical coeliac disease served as controls. RESULTS: Of the 41 patients, 5 in the challenge group and 6 in the gluten-free diet group were clinically gluten sensitive; all 11 had HLA DQ2 or DQ8. Ten of these 11 patients showed transglutaminase 2-targeted mucosal IgA deposits, which were dependent on gluten consumption. Minimal IgA deposits were seen in only 3 out of 30 patients with suspected coeliac disease without any clinically detected gluten dependency. The deposits were found in all classical coeliac patients and in none of the non coeliac control subjects. CONCLUSIONS: Clinically pertinent coeliac disease exists despite normal small-bowel mucosal villous architecture. Mucosal transglutaminase 2-specific IgA deposits can be utilized in detecting such patients with genetic gluten intolerance. PMID- 16036510 TI - Essential fatty acid depletion in children with inflammatory bowel disease. AB - OBJECTIVE: Children with inflammatory bowel disease (IBD) suffer from malabsorption and malnutrition and therefore may be at risk of developing polyunsaturated fatty acid (PUFA) deficiency. The aim of this study was to investigate PUFA status in children with IBD and the possible relationship to disease activity and nutritional status. MATERIAL AND METHODS: We assessed the fatty acid composition of plasma phospholipids (%wt/wt) of 21 children aged 5.5 18 years with IBD (ulcerative colitis, 15; Crohn's disease, 6) with mild or moderate disease activity. The clinical symptoms and biochemical indices of disease activity and nutritional status (lean and fat body mass, Hb, albumin serum conc.) were also determined. RESULTS: The patients had lower phospholipid PUFAs than 13 healthy, aged-matched controls (25.8+/-5.2 versus 34.2+/-5.7, M+/ SD, p<0.001), mainly due to lower values of linoleic acid (18:2n-6, 14.0+/-3.8 versus 18.3+/-4.3, p<0.01) and its major metabolite arachidonic acid (20:4n-6, 5.3+/-2.0 versus 9.3+/-1.9, p<0.0001). There were also higher values of a linolenic acid (18:3n-3, 0.3+/-0.4 versus 0.2+/-0.1, p<0.01) while the long-chain n-3 PUFA-eicosapentaenoic and docosahexaenoic acids were normal. Total n-6 PUFA correlated inversely to erythrocyte sedimentation rate (p<0.01), seromucoid (p<0.05) and positively to Hb concentration (p<0.01). CONCLUSIONS: Children with inflammatory bowel disease have a high risk of n-6 PUFA depletion, which is related to disease activity. PMID- 16036511 TI - Amelioration of experimental colitis by Na-H exchanger-1 inhibitor amiloride is associated with reversal of IL-1ss and ERK mitogen-activated protein kinase. AB - OBJECTIVE: Na-H exchanger-1 (NHE-1) is induced in experimental colitis. It has not yet been established whether its inhibition ameliorates colitis. The effects of amiloride, an inhibitor of NHE-1, on colitis were examined in this study. Levels of mitogen-activated protein (MAP) kinases ERK, p38 and interleukin 1ss which participate in intestinal inflammation were also examined in the colonic smooth muscle of rats with colitis. MATERIAL AND METHODS: Colitis was induced in Sprague-Dawley male rats by intrarectal administration of trinitrobenzenesulphonic acid (TNBS) and treated daily with amiloride (3, 5, and 10 mg/kg b.w. (body-weight), orally) starting 1 h before induction of colitis. The animals were sacrificed on day 5 post-TNBS. Controls received phosphate buffered saline in a similar manner. RESULTS: The highest dose of amiloride (10 mg/kg) was lethal. The lowest dose (3 mg/kg) was tolerated and was used in this study. Amiloride significantly reversed the colitis-reduced contractility and induction of MPO activity, NHE-1, IL-1ss and ERK, but not of p38 in inflamed colonic smooth muscle. Splenomegaly, increased colonic mass and decreased sodium pump activity were significantly reversed by amiloride treatment. There was no recovery of b.w. loss in the treated colitic animals. Urine output was increased, whereas food and water intake remained unchanged following amiloride treatment. CONCLUSIONS: These findings suggest that the beneficial effects of NHE-1 inhibition in experimental colitis are mediated through IL-1ss and ERK MAP kinase. PMID- 16036512 TI - Endoscopic biopsies in Ussing chambers evaluated for studies of macromolecular permeability in the human colon. AB - OBJECTIVE: Studies of mucosal permeability to protein antigens in humans are limited to in vitro techniques. The use of surgical specimens for such studies has major shortcomings. Endoscopic biopsies in Ussing chambers have been introduced as a means of studying secretion and transepithelial permeability, but have not been evaluated for studies of protein antigen uptake in human intestine. MATERIAL AND METHODS: Standard forceps biopsies from the sigmoid colon of 24 healthy volunteers were mounted in Ussing chambers with an exposed tissue area of 1.76 mm2. 51Cr-EDTA (paracellular probe) and horseradish peroxidase (HRP; 45 kDa protein antigen) were used as permeability markers. Mucosal permeability, electrophysiology, histology and energy contents of the biopsies were studied over time. To evaluate the ability of the technique to detect permeability changes, the mucosa was modulated with capric acid, a medium-chain fatty acid, known to affect tight junctions. RESULTS: In the Ussing chamber the mucosal biopsies were viable for 160 min with stable levels of ATP and lactate, and only minor changes in morphology. Steady-state permeability with low variability was seen for both markers during the 30-90 min period. Exposure to capric acid induced a rapid decrease in short-circuit current (Isc) and a slower reversible decrease in transepithelial resistance (TER), as well as an increased permeability to 51Cr-EDTA and HRP. CONCLUSIONS: Endoscopic biopsies of human colon are viable in Ussing chambers and are reliable tools for studies of mucosal permeability to protein antigens. The technique offers a broad potential for studies of mucosal function in the pathophysiology of human gastrointestinal diseases. PMID- 16036513 TI - Gastrin-17 specific antibodies are too specific for gastrinoma diagnosis but stain G-cells. PMID- 16036514 TI - Single or multiple perforations with varying locations as a complication of intestinal Behcet's disease: report of three cases. AB - Intestinal ulcers in Behcet's disease (BD) tend to cause perforation with significant morbidity. The optimal surgical procedure in such cases is controversial and the postoperative period can be eventful with an unpredictable course. We report three cases of perforations with varying locations in three patients with long-standing Behcet's disease. Two patients required two and one patient required four operations. It is emphasized that the clinician must be alert in a patient with BD when abdominal symptoms accompany the clinical picture. As soon as the diagnosis is reached, surgical intervention with limited resection must be performed. Endoscopic examination and careful medication play major roles in the follow-up. PMID- 16036515 TI - Severe cytomegalovirus enterocolitis after standard chemotherapy for non Hodgkin's lymphoma. AB - Reports of cytomegalovirus (CMV) colitis mainly concern patients with immunocompromisation resulting from, among others, HIV infection, allogeneic bone marrow transplantation and solid organ transplantation. CMV colitis rarely occurs during standard chemotherapy for non-Hodgkin's lymphoma (NHL). An unusual case of CMV enterocolitis in a 62-year-old patient is reported. After a first course of salvage chemotherapy for NHL, diffuse erosions and sloughing mucosa were seen throughout the large bowel. The final diagnosis was based on histological findings. Although ganciclovir and foscarnet are effective for CMV viremia, their use in the treatment of severe diarrhea in our patient did not result in improvement for one week, whereas concomitant use of octreotide led to rapid improvement. Octreotide may therefore be an effective agent for severe colitis. PMID- 16036516 TI - A patient with cerebral Whipple's disease and a stroke-like syndrome. AB - The central nervous system (CNS) may be affected in up to 50% of patients with Whipple's disease and this can occur even with little or no gastrointestinal involvement. We describe a 63-year-old patient in whom CNS involvement with Whipple's disease had the clinical and imaging features of a brain infarction. Treatment with aspirin and ceftriaxone followed by trimethoprim-sulfamethoxazole resulted in a good neurological recovery and complete remission of the malabsorption syndrome. Cerebral Whipple's disease resembling a stroke syndrome has so far been reported in only two other patients and in both cases it represented the first presentation of the disease. Arterial or arteriolar fibrosis, thrombosis and thickening associated with the inflammation of adjacent brain parenchyma and leptomeninges, and cerebral vasculitis caused by the hematogenous spread of Tropheryma whippelii to the brain may all be important triggers of brain infarction in patients with Whipple's disease. Our case report highlights the important point that cerebral Whipple's disease with the features of a stroke syndrome, if recognized early and treated aggressively with antibiotics, could have a favorable course with no long-term disability sequelae. PMID- 16036517 TI - Asynchronous sclerosing pancreato-cholangitis. PMID- 16036518 TI - Decreased mitochondrial oxidative capacity in hereditary tyrosinemia type 1. PMID- 16036519 TI - Ultrasonography for surgeons. PMID- 16036520 TI - Argon plasma coagulation ablation of Barrett's oesophagus. PMID- 16036521 TI - Occurrence and future history of oesophageal symptoms in an urban Swedish population: results of a questionnaire-based, ten-year follow-up study. AB - OBJECTIVE: To evaluate the occurrence and changes over time of upper gastrointestinal symptoms in adult subjects randomly recruited from the general population. MATERIAL AND METHODS: A questionnaire recording of upper gastrointestinal symptoms with a 10-year follow-up was carried out in a random sample of the adult population of Goteborg. The original sample, from 1986, comprised 441 subjects within the age span of 20-79 years. In total, 337 subjects participated in the index assessment, of which 197 (95 F) subsequently answered the second questionnaire 10 years later. RESULTS: A majority of the responders (159/197, 83%) reported no change in their global assessment of symptoms. Neither the point prevalence nor the severity for any symptom changed significantly over time. The prevalence of heartburn and acid regurgitation was 25 and 22%, respectively, and the corresponding yearly incidence was estimated to be 1.8 and 1.2%. The main predictor for the occurrence of symptoms of moderate to severe degree at follow-up was the presence of the same symptom 10 years previously. Age or gender per se did not significantly influence the occurrence of symptoms. Changes in body mass index and anti-reflux medication co-varied with the severity of the symptoms heartburn and acid regurgitation. CONCLUSIONS: Oesophageal symptoms are common in the adult Swedish population and seem to be fairly stable over time. PMID- 16036522 TI - Gastroesophageal reflux disease in children: association between symptoms and pH monitoring. AB - OBJECTIVE: The prevalence of symptoms associated with gastroesophageal reflux disease (GERD) in patients with abnormal results of pH monitoring has been investigated in adults and infants. A questionnaire suitable for children between 7 and 16 years of age has been proposed, but this tool has so far not been validated. In the present study the items of the questionnaire are validated against results from an esophageal 24-h study of pH. MATERIAL AND METHODS: Ninety nine children aged from 7 to 16 years referred from two outpatient clinics for suspected GERD completed the 7-point questionnaire regarding symptoms during the week prior to a pH study. The frequency of symptoms was investigated in patients with abnormal versus normal pH (reflux index >/< 5.0). A group of healthy children (n = 284) served as controls to estimate the frequency of symptoms in the normal population. RESULTS: It was found that 37/99 (37%) of patients had an abnormal pH study result. Regurgitation/vomiting yielded the best symptom discrimination, and was reported by 46% with abnormal versus 24% with normal pH study results (p = 0.029). A weighted score including the five best discriminating symptoms was positive in 75% versus 44% (OR 3.78, CI 1.52-9.37, p = 0.006). In a comparison of children with abnormal pH studies and healthy controls, a correct diagnosis based on five symptoms could be obtained in 75% and 94%, respectively. CONCLUSIONS: A relatively weak association was found between reflux symptoms and a positive pH study in 7-16-year-old children referred for pH monitoring. Thus, the questionnaire is not a diagnostic tool, and its potential use is limited to epidemiological studies. PMID- 16036523 TI - Th response to Helicobacter pylori differs between patients with gastric ulcer and duodenal ulcer. AB - OBJECTIVE: Helicobacter pylori (H. pylori) infection induces both gastric (GU) and duodenal ulcers (DU). We examined whether host immunological response to H. pylori determines different disease outcomes. MATERIAL AND METHODS: Thirty-two GU and 28 DU patients infected with H. pylori, and 24 dyspeptic patients without infection were enrolled. The constituents of cellular infiltrates in biopsies from each patient were determined and lymphokines secreted by stimulated T cells were measured. Serum concentrations of IgG subclasses specific to H. pylori were measured. RESULTS: Low pepsinogen I and high pepsinogen II levels were observed in GU patients, while a high pepsinogen I level was found in DU patients. T cells predominate over other cell types in both GU and DU patients. GU patients had a higher number of T cells (p < 0.01) and lower plasma cells (p < 0.05) than those in DU patients. T cells from GU patients produced greater amounts of IFN-gamma and less IL-4 than those in DU patients (p < 0.01). GU patients had a higher serum level of IgG2 specific to H. pylori than that in DU patients (p < 0.01). CONCLUSIONS: Th response by gastric T cells in GU patient was more polarized to Th1 as compared with that in DU patients, suggesting that a distinct immune response to H. pylori induces different disease outcomes. PMID- 16036524 TI - Plasma acylated ghrelin levels correlate with subjective symptoms of functional dyspepsia in female patients. AB - OBJECTIVE: Ghrelin is a brain-gut peptide that is mainly secreted from gastric endocrine cells (X/A like cells). In addition to promoting growth-hormone release and appetite, ghrelin also affects gastric motility and secretion. Circulating ghrelin levels are related to appetite and energy balance. Functional dyspepsia (FD) is a disorder characterized by the presence of chronic or recurrent symptoms of upper abdominal pain or discomfort. Although no known specific organic abnormalities are present in FD, abnormalities in gastrointestinal motility and sensitivity are thought to play a role in a substantial subgroup of patients. In addition, some patients also suffer from anorexia and body-weight loss. To investigate the role of ghrelin in the pathophysiology of FD, circulating ghrelin levels in affected patients were measured. MATERIAL AND METHODS: Eighteen Japanese female patients with functional dyspepsia and 18 healthy volunteers were recruited for the study. Acylated and desacyl forms of ghrelin were measured using commercially available enzyme-linked immunosorbent assay kits. RESULTS: Although plasma levels of acylated or desacyl ghrelin were not significantly different between healthy subjects and FD patients, plasma acylated, but not desacyl ghrelin, levels were correlated with a subjective symptom score in FD patients. In addition, the ratio of acylated to desacyl ghrelin (A/D ratio) was correlated strongly with acylated, but not desacyl, ghrelin levels. CONCLUSIONS: The correlation of circulating acylated ghrelin levels with the subjective symptom score and the A/D ratio in FD patients suggest that acylated ghrelin may play a role in the pathophysiology of FD. PMID- 16036525 TI - Effect of chemotherapy on circulating gastrointestinal hormone levels in ovarian cancer patients: relationship to nausea and vomiting. AB - OBJECTIVE: The introduction of 5-HT3 receptor antagonists greatly reduced the problems associated with nausea and vomiting immediately after cancer chemotherapy. However, delayed nausea and vomiting is still a major problem and the underlying mechanism is obscure. MATERIAL AND METHODS: We studied the effect of cisplatin-containing combination chemotherapy in 14 ovarian cancer patients on the levels of gastrin and a panel of other hormones as well as glucose and prostaglandin F2a. Blood samples were obtained once daily in the morning before chemotherapy and for 4 days after chemotherapy. RESULTS: Concentrations of many hormones including gastrin were generally high. A pronounced increase in plasma insulin levels occurred on the day after chemotherapy accompanied by a modest increase in plasma glucose concentrations. Minor increases were observed for gastrin, oxytocin and prostaglandin F2a. In contrast, a transient decrease after chemotherapy was observed for motilin. Plasma cortisol decreased markedly after chemotherapy as expected since betamethasone was given as an antiemetic prophylaxis. Certain trends concerning the relationship between some hormones and nausea and vomiting were noted. A high plasma gastrin concentration before chemotherapy was related to delayed vomiting. Relative day-to-day variability of cholecystokinin tended to correlate positively with delayed nausea, whereas an inverse relationship was observed for gastrin variability. CONCLUSIONS: Changes in hormone plasma levels were found but only few could be distinguished as possible mediators of delayed nausea and vomiting. PMID- 16036526 TI - Kinetics of mucosal granulocyte activation after gluten challenge in coeliac disease. AB - OBJECTIVE: To elucidate the dynamics of the rectal inflammatory response to rectal gluten challenge in coeliac disease by measuring inflammatory mediators released by activated neutrophils, eosinophils and mast cells/basophils. MATERIAL AND METHODS: The release of myeloperoxidase (MPO), eosinophilic cationic protein (ECP) and histamine was measured continuously during the early challenge period (3-6 h after gluten challenge) in coeliac patients (n = 9) and healthy controls (n = 5). A segmental perfusion technique was used to carry out this part of the study. Another method, the mucosal patch technique, was used to enable studies of the late challenge period (5-48 h after gluten challenge) in coeliac patients (n = 10) and healthy controls (n = 15). RESULTS: During the early challenge period the MPO levels began to increase as early as 3 h after challenge and increased progressively (p < 0.001) during the next 3 h. A decline in MPO levels was seen 15 h after challenge and another phase of increasing levels at 24 h. The MPO values declined 48 h after challenge but still remained significantly increased (p < 0.05). The ECP levels started to increase 4 h after challenge and increased progressively during the next 2 h (p < 0.05). The ECP kinetics during the late challenge period was similar as for MPO but the relative increase in ECP was more modest. No increase in histamine was found except in one patient who had a transient, early increase of histamine (3-5 h after challenge). No signs of inflammatory reaction to gluten were seen in the controls. CONCLUSIONS: There is a pronounced neutrophil activation in coeliac patients after rectal gluten challenge. This activation is apparent 4 h after challenge and remains for at least 48 h. A more modest eosinophil activation defined by ECP levels starts 1-2 h later and also remains for at least 48 h. The biphasic pattern of MPO and ECP after challenge suggests a biphasic inflammatory reaction. PMID- 16036527 TI - Changes in the expression and distribution of the inducible and endothelial nitric oxide synthase in mucosal biopsy specimens of inflammatory bowel disease. AB - OBJECTIVE: The role of nitric oxide (NO) in the pathophysiology of inflammatory bowel disease (IBD) is controversial. The aim of this study was to investigate the expression and localization of nitric oxide synthase isoforms (iNOS, eNOS) in IBD colonic mucosa. MATERIAL AND METHODS: Forty-four patients with IBD (24 ulcerative colitis (UC), 20 Crohn's disease (CD) and 16 controls) were investigated by colonoscopy. iNOS and eNOS in tissue sections was demonstrated by histochemistry (NADPH-diaphorase reaction) and immunohistochemistry. Cell type analysis and quantitative assessment of the iNOS immunoreactive (IR) cells and densitometry of iNOS in immunoblots were also performed. RESULTS: iNOS-IR cells were significantly numerous in inflamed mucosa of UC (64+/-4 cells/mm2) than in CD (4+/-2 cells/mm2). iNOS-IR/CD15-IR cells showed significant elevation in inflamed (i) versus uninflamed (u) UC mucosa (UCu 8+/-3%, UCi 85+/-10%) In CD, the percentage of iNOS-IR/CD68-IR cells was lower in inflamed sites (CDu 23+/-8%, CDi 4+/-3%). Immunoblot of biopsies revealed significant elevation of iNOS in active UC compared with uninflamed sites, whereas in CD no significant changes were detected. Differences were observed in eNOS and endothelial marker CD31 immunoreactivity. In patients with UC and in controls the ratios of eNOS/CD31-IR vessels were 82.3% and 92.0% respectively, whereas in CD the ratio was 8.3% with a concomitantly significant increase of CD31-IR vessels. The distribution and morphological characteristics of the NOS-IR inflammatory cells and endothelia were similar to those showing NADPH-diaphorase reactivity. CONCLUSIONS: Differences observed in the expression and distribution of NOS isoforms in immune and endothelial cells may contribute to better understanding of the structural and physiological changes in UC and CD. PMID- 16036528 TI - Tooth loss is associated with increased risk of gastric non-cardia adenocarcinoma in a cohort of Finnish smokers. AB - OBJECTIVE: Tooth loss has been associated with upper gastrointestinal cancer in several studies, but only one previous study used prospectively collected data. The importance of confounding by Helicobacter pylori has not previously been addressed. The objective was to determine the association between tooth loss and upper gastrointestinal cancer in the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study cohort and to determine the importance of potentially confounding dietary factors or H. pylori seropositivity. MATERIAL AND METHODS: A prospective cohort study with 29,124 subjects included 49 esophageal squamous cell carcinomas, 66 esophageal/gastric cardia adenocarcinomas, and 179 gastric non-cardia adenocarcinomas occurring between 1985 and 1999. Cox proportional hazards models adjusted for age and education were used to estimate hazard ratios (HRs) and 95% CIs. Odds ratios and 95% CIs were calculated with and without adjustment for H. pylori seropositivity in a nested case-control group to determine whether H. pylori confounded the association between tooth loss and gastric cancer. RESULTS: Tooth loss significantly increased the hazard ratio for gastric non-cardia cancer, the HR (95% CI) for edentulous subjects versus those with < 10 teeth lost was 1.65 (1.09, 2.49, respectively). No statistically significant associations were found between tooth loss and esophageal squamous cell carcinoma or esophageal/gastric cardia adenocarcinoma. Confounding by dietary factors, tobacco smoking, or H. pylori did not explain these results. CONCLUSIONS: Tooth loss was associated with increased risk of gastric non-cardia cancer, but not esophageal squamous cell carcinoma or esophageal/gastric cardia adenocarcinoma in this Finnish cohort. PMID- 16036529 TI - Long-term follow-up of interferon monotherapy in 454 consecutive naive patients infected with hepatitis C virus: multi-course interferon therapy may reduce the risk of hepatocellular carcinoma and increase survival. AB - OBJECTIVE: The long-term effects of multi-course interferon (IFN) monotherapy in patients infected with hepatitis C virus (HCV) are still unclear. MATERIAL AND METHODS: To evaluate the effects of multi-course IFN on hepatocarcinogenesis and survival, a follow-up study was conducted comprising 454 consecutively recruited non-cirrhotic naive patients infected with HCV, who had received IFN monotherapy between 1987 and 1992. The median follow-up was 11.3 years. RESULTS: A sustained response (SR) after the first IFN was achieved by 152 patients (33.5%) (Group A). Of 302 patients (66.5%) with non-SR after the first IFN, 130 patients (28.6%) did not receive additional IFN (Group B), and the remaining 172 patients (37.9%) received multi-course IFN monotherapy (Group C). With regard to hepatocarcinogenesis and survival rates for liver-related deaths, Groups A and C both showed significantly better long-term clinical outcome than Group B (p < 0.001; log-rank test). Three independent factors were identified by multivariate analyses (fibrosis stage 3, Group B, and age > or = 50) for all patients and two factors (fibrosis stage 3 and age > or = 50) for Group C associated with hepatocarcinogenesis. With regard to hepatocarcinogenesis rates according to the mean alanine aminotransferase (ALAT) levels during the IFN-free period in Group C, significantly higher rates were noted in patients with ALAT levels above 1.5 x the upper normal limit (17.6%) than those below the limit (0%) (p < 0.05). CONCLUSIONS: Multi-course IFN monotherapy reduces the risk of hepatocarcinogenesis and increases survival, and low ALAT levels during the IFN free period are associated with lower hepatocarcinogenesis rates in multi-course IFN. PMID- 16036530 TI - Prevalence and determinants of exocrine pancreatic insufficiency among older adults: results of a population-based study. AB - OBJECTIVE: The prevalence and main determinants of exocrine pancreatic insufficiency were investigated in a large population-based sample of older adults by measuring pancreatic elastase-1 in stool. MATERIAL AND METHODS: The study comprised 914 participants aged 50 to 75 years recruited by their general practitioner during a general health examination. All participants and their physicians were asked to fill out a standardized questionnaire which contained information on socio-demographic and lifestyle factors as well as medical history. Native stool was examined for pancreatic elastase-1 with a commercially available ELISA (ScheBo Tech, Giessen, Germany). RESULTS: Overall, 524 women and 390 men aged 50 to 75 years (mean age 61.9 years) were included in the analysis. In total, 105 (11.5%) of the 914 subjects showed signs of exocrine pancreatic insufficiency (EPI) with =200 microg elastase-1/g stool, and 47 (5.1%) subjects showed signs of a severe exocrine pancreatic insufficiency (SEPI, < 100 microg elastase-1/g stool). There was a clear increase in EPI with age. Patients taking angiotensin-converting enzyme (ACE) inhibitors had a lower prevalence than subjects without this medication; these associations persisted after adjustment for covariates. CONCLUSIONS: Prevalence of EPI increases with age and seems to be tentatively higher in men than in women. However, smoking seems to be an independent risk factor for EPI and SEPI whereas ACE-inhibitor intake might be a protective factor. The latter finding may even point to new options in the treatment of chronic pancreatitis. PMID- 16036531 TI - First-time endoscopy and use of antisecretory medication: a population-based cohort study. AB - OBJECTIVE: Endoscopy is the gold standard for diagnostic evaluation of upper gastrointestinal symptoms. The relation between endoscopy and use of antisecretory medication on a population level is unknown. The aim of this study was to describe development in the number of patients undergoing first-time endoscopies and their use of antisecretory medication. MATERIAL AND METHODS: Data on the use of endoscopies and antisecretory medication (H2 blockers and proton pump inhibitors) were extracted from five population-based databases and included all citizens in Funen County (population 470,000) who had first-time endoscopies between 1993 and 2002. RESULTS: A total of 27,829 first-time endoscopy patients were identified. In 2002 the number of first-time endoscopies was 5.6/1000 persons. The proportion that had redeemed prescription(s) on antisecretory medication the last year before endoscopy increased from 33% (1095/3286) in 1993 to 41% (1012/2445) in 2002 (p = 0.000). Following endoscopy, average use of antisecretory medication increased by 90 defined daily doses (DDD)/patient/year (95% CI 84-96) in patients with oesophagitis (N = 4850), by 59 DDD/patient/year (95% CI 54-64) in peptic ulcer patients (N = 4373) and by 18 DDD/patient/year (95% CI 16-20) in patients with normal endoscopies (N = 16,400). CONCLUSIONS: An increasing proportion of patients are treated with antisecretory medication before endoscopy. Following endoscopy, use of antisecretory medication increases irrespective of the diagnostic findings. PMID- 16036532 TI - ECG changes during endoscopic retrograde cholangio-pancreatography and coronary artery disease. AB - OBJECTIVE: Myocardial ischaemia has been described during endoscopic retrograde cholangio-pancreatography (ERCP), but the pathogenesis remains unclear. The aim of the present study was to evaluate whether coronary artery disease was present in patients with ST-segment changes during ERCP. MATERIAL AND METHODS: Forty patients were monitored with a Holter tape recorder during ERCP. Patients with ST segment deviation during ERCP subsequently underwent a standard exercise ECG test. RESULTS: Twelve patients developed signs of myocardial ischaemia during ERCP (30%) and 9 had concomitant tachycardia. None had a cardiac history or cardiorespiratory symptoms. Ten of the 12 patients did an exercise test and one patient developed silent ischaemia. Subsequent coronary angiography showed no evidence of coronary artery disease. CONCLUSIONS: No signs of existing coronary artery disease were found in patients developing ST deviation during ERCP when evaluated with a 12-lead exercise ECG test. Further studies should evaluate other mechanisms responsible for myocardial ischaemia during ERCP. PMID- 16036533 TI - Ultrasound may decrease the emergency surgery rate of incarcerated inguinal hernia. AB - OBJECTIVE: Manual reduction is the standard procedure for incarcerated inguinal hernia reduction. The role of ultrasound in incarcerated inguinal hernia reduction has not been defined. The aim of this study was to determine whether ultrasound can improve the ability to reduce incarcerated inguinal hernia safely when manual reduction fails and thereby decrease the emergency surgery rate. MATERIAL AND METHODS: Between January 1994 and December 2003, 112 adult patients with incarcerated inguinal hernias were admitted to a university medical center and classified into two groups. Group I consisted of 61 patients who received emergency surgical reduction after the failure of two attempts at manual reduction. In Group II, comprising 51 patients, ultrasound-guided reduction was performed when two attempts at manual reduction failed. Emergency surgical reduction was scheduled if both ultrasound-guided reduction and two attempts at manual reduction failed. The difference in emergency surgery rates between groups I and II was compared. RESULTS: In group I, manual reduction was successful in 55 cases (90.2%) and 6 patients underwent emergency surgery. In group II, manual reduction was successful in 45 cases, ultrasound-guided reduction in 4 cases, 1 reduction was avoided after ultrasonic examination, and 1 patient underwent emergency surgery. The emergency surgery rates in groups I and II were 9.8% and 2.0%, respectively. CONCLUSIONS: Ultrasound can improve the ability to reduce incarcerated inguinal hernia safely when manual reduction fails and may decrease the rate of emergency surgery. PMID- 16036534 TI - Clinical relevance of small-bowel findings detected by wireless capsule endoscopy. AB - OBJECTIVE: Capsule endoscopy is becoming known as a valid tool for identifying sources of obscure gastrointestinal (GI) bleeding. Fewer data are available about its clinical value for other indications. MATERIAL AND METHODS: Sixty patients (31 F, mean age 47 years, range 14-80 years) with no signs of overt GI bleeding were investigated by Given M2A video capsule for suspected small-bowel disease. The main clinical features were: iron deficient anemia (20), abdominal pain (12), chronic diarrhea (9), malabsorption and weight loss (7), Crohn's disease (CD) (5), and familial adenomatous polyposis (3). Three patients underwent wireless endoscopy for suspected GI neoplasm and one for portal thrombosis. RESULTS: Complete vision of the small bowel was achieved in 55 patients. No small-bowel lesions were identified in 17 patients, but 5 of them had gastric abnormalities. Small-bowel abnormality was found in 38 patients. Lesions compatible with CD were found in 14 patients, diffuse or patchy enteropathy in 7 and polyps in 6. Actively bleeding lesions were detected in 6 patients and potential bleeding sources in 5. Capsule endoscopy had an overall diagnostic yield of 62%. In particular, three small-bowel malignancies were detected and 9 patients received a better definition of their already-known pathology. However, further endoscopies were needed in 10 patients to obtain a diagnosis. One patient, diagnosed with ileal CD, underwent surgery, as the capsule remained trapped in a stricture. CONCLUSIONS: Wireless endoscopy effectively visualizes small-bowel abnormalities even though more accurate selection of the patients is needed in order to optimize its diagnostic efficacy. PMID- 16036535 TI - Acupuncture accelerates delayed gastrointestinal transit after abdominal surgery in conscious rats. PMID- 16036536 TI - Recurrent cholangitis associated with biliary sludge and Phrygian cap anomaly diagnosed by magnetic resonance imaging and magnetic resonance cholangiopancreatography despite normal ultrasound and computed tomography. AB - A 31-year-old woman presented with a one and half years' history of intermittent right upper quadrant (RUQ) pain, high fever and severely painful, warm and reddish swollen skin lesions on the fingers. Acute attack resolution occurred within 2 weeks after treatment with non-specific antibiotics. Low-grade fever (around 37.5 degrees C) and less painful swellings continued for 6 months after each attack. Abdominal ultrasound and computed tomography (CT) scans did not show any abnormality during the attacks. Biopsy of the skin lesions after the second attack revealed lymphocytic vasculitis. All laboratory studies including rheumatologic serology panel were normal. One month after the complete resolution of the second attack, the patient was observed to have high fever, the same skin lesions on the fingers as at the initial stage, nausea and marked abdominal pain in the RUQ. Routine laboratory studies including complete blood count, liver function tests and serum amylase and lipase levels were normal. An abdominal CT scan revealed a slight thickening of the gallbladder wall (3.9 mm). Two weeks later, abdominal magnetic resonance imaging (MRI) and magnetic resonance cholangiopancreatography (MRCP) were performed because of persistent abdominal pain. They revealed both biliary tract and pancreatic gland alterations consistent with past cholangitis and pancreatitis with coexisting Phrygian cap anomaly and biliary sludge on the neck of the gallbladder. PMID- 16036537 TI - Mucosal blood flow in pouchitis. PMID- 16036543 TI - Individually based preventive medical recommendations - are they sustainable and responsible? A call for ethical reflection. PMID- 16036544 TI - Quality of life of patients with diabetes mellitus. An overview of research in primary health care in the Nordic countries. AB - OBJECTIVE: A review of articles on health-related quality of life (HRQoL) among diabetes patients in primary health care in the Nordic countries. DESIGN: Search in PubMed and related sources. Used MeSH terms were: "Diabetes Mellitus", "Quality of Life", "Primary Health Care", and "Scandinavia" or "Denmark", "Finland", "Iceland", "Norway", or "Sweden". RESULTS: In total, 19 relevant articles were found, 9 from Finland and 10 from Sweden. HRQoL questionnaires used were from the Medical Outcomes Study, i.e. the SWED-QUAL in seven, SF-20 in four and SF-36 in one, NHP in two, GQL in two, GHQ in one, and own questionnaires in two. HRQoL was moderately affected in diabetic patients, with macrovascular diseases, especially coronary heart disease, and non-vascular diseases as the most consistently found and strongest predictors. Weaker predictors were microvascular complications, age, sex, metabolic level, and education. Other groups at higher risk of worse HRQoL were diabetes patients with psychiatric disorders, especially depression, and foreign-born patients. Good continuity of care was associated with better HRQoL. CONCLUSION: Diabetes affects the HRQoL through macrovascular complications and associated non-vascular comorbidity, and also by adding to the total burden of disease. Future research areas include transcultural and ethnic aspects, and effects of lifestyle interventions. PMID- 16036545 TI - Reduction of high cholesterol levels associated with younger age and longer education in a primary health care programme for cardiovascular prevention. AB - OBJECTIVE: To study possible social predictors for reduction of hyperlipidaemia in subjects offered lifestyle intervention in primary health care after an opportunistic screening. SETTING: Primary health care in Sollentuna, Sweden. DESIGN: Follow-up study of changes in high lipid levels in men and women aged 20 60 years participating in a voluntary screening and cardiovascular prevention programme. SUBJECTS AND MAIN OUTCOME MEASURES: A total of 1904 individuals had a follow-up visit registered after a mean of 466 days. Men and women with raised lipid levels (serum cholesterol = 6.5 mmol/l, and/or triglycerides = 2.3 mmol/l) at baseline were compared with normolipidaemic participants. Data on social characteristics such as education, occupation, marital status, and income were collected from national censuses. Associations between socioeconomic factors and changes in lipid levels were studied. RESULTS: Men and women with hyperlipidaemia were generally (p < 0.001) older (men 6-8 years, women 8-10 years) and less educated than normolipidaemic subjects. Significant predictors for reducing hypercholesterolaemia were younger age, OR 0.97 (0.95-1.00) for increasing age, and longer education, OR 0.47 (0.24-0.91) for low education (<9 years). Foreign born subjects were more likely to achieve a high success rate in reducing hypercholesterolaemia, OR 3.43 (1.00-11.8), than the Swedish-born. No significant predictors were detected for reduction of high triglyceride levels. CONCLUSION: A successful reduction of high cholesterol levels was associated with younger age and longer education in a primary health care-based programme for cardiovascular prevention. PMID- 16036546 TI - Low-budget method for lifestyle improvement in primary care. Experiences from the Goteborg Health Profile Project. AB - OBJECTIVE: To describe a self-administered preventive tool dealing with risk factors for cardiovascular disease and its effect with special reference to the question: did the project involve persons most in need of lifestyle changes? DESIGN: Screening questions offered to consecutive patients and followed by a self-administered health profile, intervention, and follow-up. Setting. Primary healthcare area of Askim, Sweden. SUBJECTS: Men and women between 18 and 65 years of age visiting GPs for acute disorders or planned visits during a three-month period were offered screening questions and, if wanted, a health profile. MAIN OUTCOME MEASURES: Participation rates, effects on lifestyle factors. RESULTS: There was an overrepresentation of subjects with a less favourable lifestyle among those who asked for the health profile. There was good agreement for all variables between self-estimation in screening questions and grading in the basal health profile. Lifestyle improvement was observed for dietary habits, physical activity, and mental stress at the one year follow-up. CONCLUSIONS: The results indicate that a relevant selection of persons with a "risk profile" can be made by means of simple screening questions. The pedagogic model using self administered health profile in combination with own responsibility seems to be a tool for low-budget preventive work in primary healthcare. PMID- 16036547 TI - Comorbidity and medication load in adult asthmatics. AB - OBJECTIVE: To examine comorbidity and the medication load among asthmatics. Design A self-administered postal inquiry. SETTING: A national register-based random sample of 6000 subjects aged 16 years or older entitled to special reimbursement for anti-asthmatic medication in Finland. SUBJECTS: A total of 4690 subjects with clinically diagnosed asthma. MAIN OUTCOME MEASURES: Reporting of doctor-diagnosed chronic diseases and the number of prescription medicines used by asthmatics. RESULTS: Two-thirds of the subjects (n = 2952, 63%) reported other diseases in addition to other chronic pulmonary diseases and allergies. Musculoskeletal and cardiovascular disorders were the most common, increasing with age. Allergies were most frequent among the young asthmatics. Nearly all the subjects (n = 4444, 95%) took at least one anti-asthmatic medicine, and two out of every three (n = 3051, 65%) received other prescription medicines, most commonly cardiovascular drugs or analgesics. Some 41% (n=1938) of all the asthmatics and as many as 21% of the young adults (n = 269) were taking at least five prescription medicines concomitantly. The total medication load increased with age. CONCLUSION: The load of comorbidity and prescribed medication is heavy in adult asthmatics of all ages. Thus asthmatic patients should best be treated by GPs, while pulmonary specialists work as consultants and take care of the most severe cases. PMID- 16036548 TI - Social background, aspects of lifestyle, body image, relations, school situation, and somatic and psychological symptoms in obese and overweight 15-year-old boys in a county in Sweden. AB - OBJECTIVE: To compare lifestyle and health aspects among obese, overweight, and normal-weight 15-year-old boys living in the county of Vastmanland, Sweden. DESIGN: A cross-sectional school-based survey. Setting. All schools in the county of Vastmanland, Sweden. SUBJECTS: A questionnaire was completed anonymously by 989 boys. MAIN OUTCOME MEASURES: The relations between body mass index and social factors, eating habits, physical activity, body image, relations, school situation, use of alcohol, drugs and tobacco, somatic and psychological symptoms in boys. RESULTS: Obese boys had a significant negative outcome in 19 out of 31 items studied compared with normal-weight boys, while the overweight boys had a significant negative outcome in 9 out of 31. The obese boys reported more irregular eating habits than normal-weight boys, were less satisfied with their weight and looks, and had fewer friends. A larger proportion of the obese boys reported that they did not like school, were more absent from school, and had been exposed to more violence. They bullied their schoolmates more often. The obese boys had tried sniffing solvents and used illicit drugs more frequently than their peers. They reported more somatic and psychological symptoms as well as suicidal thoughts and attempts. CONCLUSIONS: Obese 15-year-old boys differed from overweight and normal-weight boys in lifestyle and in the frequency of somatic and psychological symptoms. Early and vigorous intervention is necessary, as they may belong to a risk group that could develop not only medically but also socially negative consequences. PMID- 16036549 TI - Justifications of physicians' choice of action. AB - OBJECTIVE: To compare choice of actions and justifying reasons among the general public, GPs and oncologists. DESIGN: A postal questionnaire using four vignettes: a healthy patient reluctant to quit smoking and one demanding an X-ray, a pulmonary cancer patient reluctant to quit smoking and one demanding immunotherapy. Subjects A random sample of the general public (n = 1000), GPs (n = 200), and oncologists (n = 200). MAIN OUTCOME MEASURES: The respondents' choice of how the physician should act (general public)/would act (physicians), and, with reference to their choice of action, their justifying reasons. RESULTS: A majority answered that the physician should/would bring up the question about smoking cessation with the reluctant patient, justified by promotion of medical benefit and protection from harm. With reference to the reluctant cancer patient every second oncologist and every third GP would not mention smoking cessation, justified by respect for self-determination and enhancement of the relationship. Facing the patient demanding a new X-ray the general public was more in favour while the physicians rejected it. One-third of the general public answered that the seriously ill patient's demand for immunotherapy should be granted and referred to respect for self-determination and enhanced relationship. A majority of the physicians rejected such a demand, justified by promotion of a fair distribution of resources. CONCLUSION: The fear of infringing the patient's right to self-determination and harming the relationship is exaggerated by physicians. They are mostly expected to bring up smoking cessation with a reluctant patient. How to manage demanding patients is more controversial. PMID- 16036550 TI - Genetic consultations in primary care: GPs' responses to three scenarios. AB - OBJECTIVE: This study investigated general practitioners' responses to three scenarios in which patients consulted regarding genetic conditions. DESIGN: Self completed postal study. Setting. Primary care in Northern Ireland. SUBJECTS: Questionnaire were distributed to all the GPs in Northern Ireland (n = 1079). A total of 541 GPs participated (50%). MAIN OUTCOME MEASURES: Responses to three scenarios in which patients consulted regarding their family history and risk of bowel cancer, breast cancer, and cystic fibrosis. RESULTS: Most GPs correctly identified the patients' risk of bowel cancer, recommended regular colonoscopy, advised lifestyle changes, and did not refer to the genetic clinic. GPs who were qualified for longer were more likely to recommend colonoscopy and less likely to advise lifestyle changes. With the breast cancer patient GPs adopted a cautious approach; most would refer to the genetic and mammography clinics. With the cystic fibrosis example, most correctly identified the patient's risk of carrying the gene, would refer to the genetic clinic, and would encourage the patient to discuss the risk with his partner. In general, doctors were unsure, but would pass on genetic information to insurance companies if requested. CONCLUSION: The study suggests that, in most cases, general practitioners correctly identify at risk individuals but there may still be some uncertainty regarding referrals. The results suggest that ways of educating GPs should be explored. Educational interventions should be linked to a greater understanding of factors involved in referral (including the influence of gender and experience). The guidelines provided to GPs in relation to the provision of genetic information to insurance companies may need to be reviewed in some countries. PMID- 16036551 TI - Uncomplicated urinary tract infections. Bacterial findings and efficacy of empirical antibacterial treatment. AB - OBJECTIVE: To assess bacterial aetiology, antimicrobial susceptibility and efficacy of empirical treatment in uncomplicated urinary tract infections and to evaluate the dipstick as a diagnostic tool. DESIGN: Prospective study. SETTING: Clinical microbiology laboratory and 17 general practice clinics in Telemark County, Norway. SUBJECTS: A total of 184 female patients between 15 and 65 years of age with symptoms of uncomplicated urinary tract infection. MAIN OUTCOME MEASURES: Results from dipstick testing (leucocyte esterase and nitrite), bacterial culture, susceptibility patterns and efficacy of empirical antibacterial therapy on symptoms. RESULTS: Significant bacteruria was detected in 140 (76%) of the 184 urines. The leukocyte esterase test was of little help in predicting culture-positive UTI. A positive nitrite test accurately predicted culture-positivity, while a negative result was ambiguous. The most common bacterium, E. coli, was found in 112 (80%) of the 140 positive urines and was predominantly sensitive to ciprofloxacin (100%), mecillinam (94%), nitrofurantoin (97%), trimethoprim (88%), and sulphonamide (81%), and to a lesser extent to ampicillin (72%). In 18 patients the causative bacterium was resistant to the therapeutic agent used; 7 of these returned to their GP with persisting symptoms while in 11 symptoms resolved without further treatment. CONCLUSION: The study confirms E. coli as the predominant cause of uncomplicated UTI. Since in the majority of cases the bacterium found was susceptible to the locally preferred antimicrobials and the patients' symptoms were cured, empiric therapy is found to be an effective practice in the study area and, by inference, in others with similar antimicrobial susceptibility patterns. PMID- 16036552 TI - GPs' reasons for "non-pharmacological" prescribing of antibiotics. A phenomenological study. AB - OBJECTIVE: To study the reasons cited by Icelandic general practitioners for their "non-pharmacological" prescribing of antibiotics. DESIGN: A qualitative interview study with research dialogues guided by the Vancouver School of doing phenomenology. SETTING: General practice. Participants A total of 16 general practitioners: 11 in the maximum variety sample and 5 in the theoretical sample. RESULTS: The most important reasons for prescribing antibiotics in situations with low pharmacological indications (non-pharmacological prescribing) were an unstable doctor-patient relationship due to lack of continuity of care, patient pressure in a stress-loaded society, the doctor's personal characteristics, particularly zeal and readiness to serve, and, finally, the insecurity and uncertainty of the doctor who falls back on the prescription as a coping strategy in a difficult situation. CONCLUSION: The causes of non-pharmacological prescribing of antibiotics are highly varied, and relational factors in the interplay between the doctor and the patient are often a key factor. Therefore, it is of great importance for the general practitioner to know the patient and to become better equipped to resist patient pressure, in order to avoid the need to use the prescription as a coping strategy. Continuity of medical care and a stable doctor-patient relationship may be seen as the core concepts in this study and the most important task for the GPs is to promote the patients' trust. PMID- 16036553 TI - Abdominal complaints in general practice: diagnoses and characteristics of patients. PMID- 16036554 TI - Global problems and global science. PMID- 16036555 TI - J.T. Baker horrors. PMID- 16036556 TI - Protein--protein interactions in the eubacterial replisome. AB - Replication of genomic DNA is a universal process that proceeds in distinct stages, from initiation to elongation and finally to termination. Each stage involves multiple stable or transient interactions between protein subunits with functions that are more or less conserved in all organisms. In Escherichia coli, initiation of bidirectional replication at the origin (oriC) occurs through the concerted actions of the DnaA replication initiator protein, the hexameric DnaB helicase, the DnaC?helicase loading partner and the DnaG primase, leading to establishment of two replication forks. Elongation of RNA primers at each fork proceeds simultaneously on both strands by actions of the multimeric replicase, DNA polymerase III holoenzyme. The fork that arrives first in the terminus region is halted by its encounter with a correctly-oriented complex of the Tus replication terminator protein bound at one of several Ter sites, where it is trapped until the other fork arrives. We summarize current understanding of interactions among the various proteins that act in the different stages of replication of the chromosome of E. coli, and make some comparisons with the analogous proteins in Bacillus subtilis and the coliphages T4 and T7. PMID- 16036557 TI - Scorpion venom peptides without disulfide bridges. AB - Several hundred disulfide-bridged neurotoxic peptides have been characterized from scorpion venom; however, only few scorpion venom peptides without disulfide bridges have been identified and characterized. These non-disulfide-bridged peptides (NDBPs) are a novel class of molecules because of their unique antimicrobial, immunological or cellular signaling activities. This review provides an overview of their structural simplicity, precursor processing, biological activities and evolution, and sheds insight into their potential clinical and agricultural applications. Based on their pharmacological activities and peptide size similarity, we have classified these peptides into six subfamilies. PMID- 16036558 TI - On the existence of cellular tocopheryl phosphate, its synthesis, degradation and cellular roles: a hypothesis. AB - The finding that alpha-tocopheryl phosphate is present in cells in small amounts, that it can be synthesized and hydrolyzed supports the hypothesis that alpha tocopheryl phosphate might be a signaling molecule. The possible pathways needed for the synthesis, hydrolysis and signaling are considered in this hypothesis as well the possible extension of this reaction to additional molecules such as tocopherols and tocotrienols. A possible mechanism of action of other tocopherol esters (succinate and maleate) is also hypothesized. PMID- 16036559 TI - The mitochondrial ribomotor hypothesis. AB - The mitochondrial ribomotor model has been proposed to explain how the balance between rRNA and mRNA in mammalian mitochondria is regulated. In this model, the interaction of the mitochondrial transcription termination factor (mTERF) with some unknown component(s), causes a loop to form in the mtDNA chain that brings the initiation and termination regions together at its base. By bringing these sites into closer proximity, the mtRNA polymerase molecules can be directly transferred from the termination site to the IH1 initiation site of the H-strand once transcription terminates. This process occurs when mTERF is phosphorylated. When unphosphorylated, transcription is initiated from the IH2 site and the polymerase reads through the mTERF-dependent termination site, resulting in the transcription of almost the entire H-strand. PMID- 16036560 TI - Effect of ribozyme against transforming growth factorbeta1 on biological character of activated HSCs. AB - Transforming growth factorbeta1 (TGFbeta1) is considered to be the principal contributor to liver fibrosis. So in this study the ribozymes against TGFbeta1 were designed. The in vitro cleavage activities of the ribozymes were assayed through incubation of (32)p-labeled target RNAs and (32)p-labeled ribozymes in different conditions. HSC-T6 cells were transfected with the eukaryotic constructs encoding ribozyme and disable ribozyme, then the stable cell clones were used to evaluate its antifibrotic characteristic through the effect of ribozyme on biological character of activated hepatic stellate cells (HSCs). The results demonstrated that two ribozymes (Rz803 and Rz1395) could cleave target RNAs into expected products effectively, Rz803 possessed better cleavage activity in vitro. Stable transfection of Rz803 into activated HSCs reduced TGFbeta1 expression in mRNA and protein level efficiently. The further studies demonstrated that Rz803 reduced deposition of collagen I, suppressed HSC proliferation, but had no effect on HSC activation in transfected HSC-T6 cells. Therefore, it indicated that Rz803 could reverse the character of activated HSCs by down-regulating TGFbeta1 expression efficiently and diminishing TGFbeta1 signaling underlying activation of hepatic stellate cells. As the consequence, it would provide a potential therapeutic approach for liver fibrosis. PMID- 16036561 TI - Differential expression of rat beta-defensins. AB - The innate immunity, utilizes a battery of broad-spectrum antibacterial cationic polypeptides (3-5 kDa) named alpha- and beta-defensins. Several beta-defensins have been isolated and shown to play a role in the defense of various tissues. Herein, we report the expression pattern of two rat beta-defensins, rBD-1 and rBD 2, in liver, kidney, lung, spleen, and brain using RT-PCR. To study polymorphism and verify gene identity, all cDNA products were sequenced. rBD-1 was expressed in the kidney, lung, brain, but not in spleen or liver, whereas rBD-2 was expressed in the lung, but not in the kidney or spleen. In addition, rBD-2 was expressed in the brain and liver. No polymorphism was found in the genes encoding rat beta-defensins. These findings demonstrate a different expression pattern for rBD-2 than what has been reported. We conclude that the rat may be a useful model to investigate the function and contribution of beta-defensins to host defense. PMID- 16036562 TI - Is there an answer? PMID- 16036563 TI - How I became a biochemist. PMID- 16036564 TI - Stochastic cell. AB - Accumulating experimental evidence of stochasticity, self-organization and abrupt non-linear transitions underlying the dynamics of cellular structure and function is increasingly more consistent with the concepts and models of phase transitions, critical phenomena and non-linear thermodynamics rather than with the conventional clockwork description of the cell. The novel emerging image of the stochastic cell suggests that familiar and convenient classico-mechanical interpretations may be limiting our ability to understand the behavior of biological systems and calls for active exploration of alternative interpretational frameworks. PMID- 16036565 TI - Viral interactions with the nuclear transport machinery: discovering and disrupting pathways. AB - Viruses have been invaluable tools for discovering key pathways of nucleocytoplasmic transport. Conversely, disruption of specific nuclear transport pathways, are crucial for the productive life cycle of some viruses. The major cellular mRNA export pathway, which uses TAP (NXF1)/p15(NXT) as receptor, was discovered as a result of TAP interaction with CTE-containing RNAs from Mason Pfizer Monkey Virus. In addition, CRM1 or exportin 1, which is a transport receptor that mediates nuclear export of proteins, snRNAs, rRNAs and a small subset of mRNAs, was discovered as an interacting partner of the Rev protein of HIV1. Viruses may disrupt the nuclear transport machinery to prevent host antiviral response. VSV Matrix (M) protein inhibits mRNA export by forming a complex with the mRNA export factor Rae1 whereas poliovirus inhibits nuclear import of proteins by probably degrading Nup62 and Nup153. Hence, this review focuses on viruses as tools and as disruptors of nucleocytoplasmic trafficking. PMID- 16036566 TI - The power and potential of doxorubicin-DNA adducts. AB - Doxorubicin (trade name Adriamycin) is a widely used anticancer agent which exhibits good activity against a wide range of tumors. Although the major mode of action appears to be normally as a topoisomerase II poison, it also exhibits a number of other cellular responses, one of which is the ability to form adducts with DNA. For adduct formation doxorubicin must react with cellular formaldehyde to form an activated Schiff base which is then able to form an aminal (N-C-N) linkage to the exocyclic amino group of guanine residues. The mono-adducts form primarily at G of 5'-GCN-3' sequences where the chromophore of the drug is intercalated between the C and N base pair. The structure of the adducts has have been well defined by 2D NMR, mass spectrometry and X-ray crystallography. The formation of these anthracycline adducts in cells grown in culture has been unequivocally demonstrated. The source of formaldehyde in cells can be endogenous, provided by coadministration of prodrugs that release formaldehyde or by prior complexation of anthracyclines with formaldehyde. Since the adducts appear to be more cytotoxic than doxorubicin alone, and also less susceptible to drug-efflux forms of resistance, they offer new approaches to improving the anticancer activity of the anthracyclines. PMID- 16036567 TI - Mechanisms of persistent NF-kappaB activation by HTLV-I tax. AB - Human T cell leukemia virus type I (HTLV-I) is the causative agent of a fatal malignancy known as adult T cell leukemia (ATL). The HTLV-I Tax protein is thought to play a significant role in the initiation and pathogenesis of HTLV-I mediated disease. Tax is a potent oncogene that deregulates cellular gene expression by persistently activating signaling pathways such as NF-kappaB. Tax activation of NF-kappaB is critical for the immortalization and survival of HTLV I-infected T cells. In this review, we describe recent insights into the mechanisms employed by Tax to activate the canonical and noncanonical NF-kappaB signaling pathways. The adaptor function of Tax appears to be a common and important mechanism for the pathological activation of both NF-kappaB pathways. PMID- 16036568 TI - Rpb4 and Rpb7: a sub-complex integral to multi-subunit RNA polymerases performs a multitude of functions. AB - Rpb4 and Rpb7, are conserved subunits of RNA polymerase II that play important roles in stress responses such as growth at extreme temperatures, recovery from stationary phase, sporulation and pseudohyphal growth. Recent reports have shown that apart from stress response, these proteins also affect a multitude of processes including activated transcription, mRNA export, transcription coupled repair etc. We propose a model that integrates the multifarious roles of this sub complex. We suggest that these proteins function by modulating interactions of one or more ancillary factors with the polymerase leading to specific transcription of subsets of these genes. Preliminary experimental evidence in support of such a model is discussed. PMID- 16036569 TI - Primary platelet adhesion receptors. AB - Thrombotic diseases such as heart attack and stroke remain a major health concern in the Western world despite existing anti-thrombotic drugs. Current studies are revealing structure-function relationships of primary platelet adhesion receptors mediating adhesion, activation and aggregation, and the molecular mechanisms underlying platelet thrombus formation. Platelet adhesion is relevant not only to thrombotic disease, but there is increasing evidence of a specific role for platelets in vascular processes such as inflammation and atherogenesis. This review focuses on recent advances in understanding the molecular basis for platelet thrombus formation, in particular the receptors, glycoprotein (GP)Ib-IX V and GPVI, that initiate platelet adhesion and activation at high shear stress. PMID- 16036570 TI - Evidence for rat organic anion transporter 3 association with caveolin-1 in rat kidney. AB - The rat organic anion transporter 3 (rOAT3) has recently been identified as the third isoform of the OAT family. The mechanisms that regulate rOAT3's functions remain to be elucidated. rOAT3 contributes for moving a number of negatively charged organic compounds between cells and their extracellular milieu. Caveolin (Cav) also plays a role as a membrane transporter. To address the relationship of these two proteins, we investigated the protein-protein interaction between rOAT3 and Cav-1. The rOAT3 mRNA and protein expression were observed in the rat kidney, and the expressions of Cav-1 mRNA and protein were also detected in the kidney. Confocal microscopy of the immuno-cytochemistry experiments using primary cultured renal proximal tubular cells showed that rOAT3 and Cav-1 were co localized at the plasma membrane. This finding was confirmed by Western blot analysis using isolated caveolae-enriched membrane fractions from the rat kidney and immuno-precipitation experimentation. When rOAT3's synthesized cRNA of rOAT3 along with the antisense oligo deoxynucleotide ofXenopusCav-1 were co-injected intoXenopusoocytes, the [(3)H] estrone sulfate uptake was significantly decreased. These findings suggest that rOAT3 and caveolin-1 share a cellular expression in the plasma membrane and Cav-1 up-regulates the organic anionic compound uptake via rOAT3 under normal physiological conditions. PMID- 16036571 TI - Is there an answer? Is there a genetic basis for human susceptibility to leprosy? PMID- 16036572 TI - How I became a biochemist. PMID- 16036573 TI - Vitamin B4. PMID- 16036574 TI - Molecular signaling aptamers for real-time fluorescence analysis of protein. AB - Aptamers are a new class of nucleic acids that are selected in vitro for binding target molecules with high affinity and selectivity. They are promising protein binding molecular probes that rival conventional antibodies for protein analysis. There have been recent advances in the development of molecular signaling aptamers that can transduce target protein binding to sensitive fluorescence signal changes. This facilitates the real time protein monitoring in homogenous solution as well as potentially in vivo. Different signaling strategies of using dual labeled aptamers based on fluorescence resonance energy transfer (FRET), one fluorophore labeled aptamers based on fluorescence anisotropy assay, or other label-free aptamers are reviewed. PMID- 16036575 TI - Yeast programmed cell death: an intricate puzzle. AB - Yeasts as eukaryotic microorganisms with simple, well known and tractable genetics, have long been powerful model systems for studying complex biological phenomena such as the cell cycle or vesicle fusion. Until recently, yeast has been assumed as a cellular 'clean room' to study the interactions and the mechanisms of action of mammalian apoptotic regulators. However, the finding of an endogenous programmed cell death (PCD) process in yeast with an apoptotic phenotype has turned yeast into an 'unclean' but even more powerful model for apoptosis research. Yeast cells appear to possess an endogenous apoptotic machinery including its own regulators and pathway(s). Such machinery may not exactly recapitulate that of mammalian systems but it represents a simple and valuable model which will assist in the future understanding of the complex connections between apoptotic and non-apoptotic mammalian PCD pathways. Following this line of thought and in order to validate and make the most of this promising cell death model, researchers must undoubtedly address the following issues: what are the crucial yeast PCD regulators? How do they play together? What are the cell death pathways shared by yeast and mammalian PCD? Solving these questions is currently the most pressing challenge for yeast cell death researchers. PMID- 16036576 TI - Transcriptional regulation by the acetylation of nonhistone proteins in humans -- a new target for therapeutics. AB - Gene expression from the dynamic chromatin template is regulated by certain key cellular players that cause post-translational modifications of both histones and nonhistone proteins. The acetyltransferases and deacetylases are two such key groups of enzymes that play crucial roles in maintaining the reversible acetylation status of histones and nonhistone proteins. Emerging evidence suggests that acetylation of nonhistone protein is equally important in the transcription regulation as the histone acetylation. Since dysfunction of HATs and HDACs leads to several diseases, aberrant acetylation of nonhistone protein is also associated with diseases. Small molecule modulators of these enzymes, which may help in maintaining the normal cellular acetylation status of these proteins, have important therapeutic implications. PMID- 16036577 TI - Stress responses in mycobacteria. AB - Mycobacterium tuberculosis is a successful pathogen that overcomes numerous challenges presented by the immune system of the host. This bacterium usually establishes a chronic infection in the host where it may silently persist inside a granuloma until, a failure in host defenses, leads to manifestation of the disease. None of the conventional anti-tuberculosis drugs are able to target these persisting bacilli. Development of drugs against such persisting bacilli is a constant challenge since the physiology of these dormant bacteria is still not understood at the molecular level. Some evidence suggests that the in vivo environment encountered by the persisting bacteria is anoxic and nutritionally starved. Based on these assumptions, anaerobic and starved cultures are used as models to study the molecular basis of dormancy. This review outlines the problem of persistence of M. tuberculosis and the various in vitro models used to study mycobacterial latency. The basis of selecting the nutritional starvation model has been outlined here. Also, the choice of M. smegmatis as a model suitable for studying mycobacterial latency is discussed. Lastly, general issues related to oxidative stress and bacterial responses to it have been elaborated. We have also discussed general control of OxyR-mediated regulation and emphasized the processes which manifest in the absence of functional OxyR in the bacteria. Lastly, a new class of protein called Dps has been reviewed for its important role in protecting DNA under stress. PMID- 16036578 TI - Collagen structure: the Madras triple helix and the current scenario. AB - This year marks the 50th anniversary of the coiled-coil triple helical structure of collagen, first proposed by Ramachandran's group from Madras. The structure is unique among the protein secondary structures in that it requires a very specific tripeptide sequence repeat, with glycine being mandatory at every third position and readily accommodates the imino acids proline/hydroxyproline, at the other two positions. The original structure was postulated to be stabilized by two interchain hydrogen bonds, per tripeptide. Subsequent modeling studies suggested that the triple helix is stabilized by one direct inter chain hydrogen bond as well as water mediated hydrogen bonds. The hydroxyproline residues were also implicated to play an important role in stabilizing the collagen fibres. Several high resolution crystal structures of oligopeptides related to collagen have been determined in the last ten years. Stability of synthetic mimics of collagen has also been extensively studied. These have confirmed the essential correctness of the coiled-coil triple helical structure of collagen, as well as the role of water and hydroxyproline residues, but also indicated additional sequence dependent features. This review discusses some of these recent results and their implications for collagen fiber formation. PMID- 16036579 TI - A novel snoRNA can direct site-specific 2'-O-ribose methylation of snRNAs in Oryza sativa. AB - Small nucleolar RNAs (snoRNAs) are a kind of noncoding RNAs, and the vast majority of snoRNAs are involved in site-specific modifications of rRNAs. A novel box C/D snoRNA called snoR124 was found inOryza sativa, and it can direct 2'-O ribose methylation of spliceosomal small nuclear RNAs (snRNAs). The snoRNA has two antisense elements, and the results of primer extensions at different dNTP concentrations provide evidence that snoR124 guide 2'-O-methylations of the C76 residue in the U4 snRNA and the T91 residue in the U5 snRNA. In addition, this snoRNA is located in a snoRNA gene cluster with another 7 snoRNAs which are identified to direct ribose methylations in rRNAs. This is consistent with the opinion that the snoRNA gene organization in plant is mainly gene cluster. The snoR124 is the first example of a snoRNA that directs modifications of RNAs other than rRNAs in plant; it will avail to get more insights into the function of snoRNAs in plant. PMID- 16036580 TI - Hydrogen peroxide-induced cell death in Arabidopsis: transcriptional and mutant analysis reveals a role of an oxoglutarate-dependent dioxygenase gene in the cell death process. AB - Hydrogen peroxide is a major regulator of plant programmed cell death (PCD) but little is known about the downstream genes from the H(2)O(2)-signaling network that mediate the cell death. To address this question, a novel system for studying H(2)O(2)-induced programmed cell death in Arabidopsis thaliana was used. The catalase inhibitor aminotriazole (AT) reduced the catalase activity and caused endogenous accumulation of hydrogen peroxide that eventually triggered cell death. Microarray analysis with a DNA chip representing 21500 genes and subsequent comparison with other PCD-related expression studies revealed a set of new H(2)O(2)-responsive genes that were highly regulated in a common fashion during different types of PCD. These included an oxoglutarate-dependent dioxygenase and various oxidoreductases, the transcription factors Zat11, WRKY75 and NAM, proteasomal components, a heterologous group of genes with diverse functions, and genes encoding proteins with unknown functions. Knockout lines of the oxoglutarate-dependent dioxygenase exhibited significantly reduced death symptoms and chlorophyll loss upon H(2)O(2)-induced cell death, indicating a role for this gene in the cell death network. PMID- 16036581 TI - Is there an answer? PMID- 16036582 TI - How I became a biochemist. PMID- 16036584 TI - The IUB(MB): 1955--2005. PMID- 16036607 TI - Scientific publishing in non industrialized countries: a pilot wireless internet project for Africa. AB - There is general agreement that the internet is the major means of future scientific communication and education. However not everybody appreciates that the development of electronic communication in industrialized societies is not matched, even to a small extent, in developing countries. Several new technologies offer the potential for developing countries to provide connectivity. Terrestrial wireless and satellite technologies offer many advantages in that they do not require installation of wire-line networks. Satellite facilities can also be installed where communication is needed, even in remote and isolated areas, rather than waiting for terrestrial networks to be extended from the cities. PMID- 16036608 TI - Dual nature of the genome: genes for the individual life and genes for the evolutionary progress of the population. AB - Biological evolution is here postulated to be driven coordinately by the products of specific evolution genes and by non-genetic elements such as the intrinsic properties of matter and random encounter with environmental factors. Evolution genes are supposed to have their own evolutionary history in which second-order selection was exerted at the population level. The products of evolution genes can act as generators of genetic variations and/or as modulators of the frequency of genetic variation. Three major natural strategies, each with a number of specific mechanisms contribute to the overall spontaneous production of genetic variants. Each of these three strategies contributes its own specific quality to genetic variation. The difficulties of experimentally investigating these strategies and a wider discussion of some of the postulates within the scientific community are outlined. Finally, the general relevance of the postulated duality of the genome for our world view is briefly mentioned. PMID- 16036609 TI - From proteins to proteomics. AB - During the second half of the 20th century, biochemistry and subsequently molecular biology blossomed into the core upon which all biological and biomedical sciences now depend. A major part of these closely related disciplines has been the study of the structure and function of proteins and the diverse biological functions that they perform. Early experimentation necessarily focused on individual entities, selected mainly for their activities, but as technology improved there developed a tendency to look at proteins as larger, interactive groups or clusters. Spurred by the recent exponential production of genomic sequence data for a rapidly increasing number of species, protein chemistry has now evolved into a new discipline, proteomics. In addition to embracing the methods and approaches that have served protein scientists well in the past, it includes, and is perhaps best defined by, high-throughput analyses based in large part on 2D gel electrophoresis, MALDI and ESI mass spectrometry and combinatorial arrays. Proteomic targets include the identification of all genome products and a mapping of their interactions and expression profiles. These hold great promise for the identification of disease markers and drug targets, but are not without their challenges and pitfalls. PMID- 16036610 TI - Bovine spongiform encephalopathy--some surprises for biochemists. AB - Bovine Spongiform Encephalopathy (BSE) is typical of the dementias that affect both animals and man; Scrapie in sheep, Creutzfeldt-Jakob disease in man. Global efforts have been made to determine the nature of the active agents in these diseases. At present the 'protein only hypothesis' of Prusiner holds. It was a surprise that a protein could per se be the active agent but other surprises for our traditional teaching of biochemistry arose. These are explained in a brief summary of our present understanding of the biochemistry of the active agents that cause the diseases. PMID- 16036611 TI - Reactive oxygen species as mediators of cellular senescence. AB - Aging has often been viewed as a random process arising from the accumulation of both genetic and epigenetic changes. Increasingly, the notion that aging is a stochastic process is being supplanted by the concept that maximum lifespan of an organism is tightly regulated. This knowledge has led to a growing overlap between classical signal transduction paradigms and the biology of aging. We review certain specific examples where these seemingly disparate disciplines intersect. In particular, we review the concept that intracellular reactive oxygen species function as signalling molecules and that oxidants play a central role as mediators of cellular senescence. PMID- 16036612 TI - From JNK to pay dirt: jun kinases, their biochemistry, physiology and clinical importance. AB - The c-Jun N-terminal kinases (JNKs) were originally identified by their ability to phosphorylate c-Jun in response to UV-irradiation, but now are recognized as critical regulators of various aspects of mammalian physiology, including: cell proliferation, cell survival, cell death, DNA repair and metabolism. JNK-mediated phosphorylation enhances the ability of c-Jun, a component of the AP-1 transcription factor, to activate transcription, in response to a plethora of extracellular stimuli. The JNK activation leads to induction of AP-1-dependent target genes involved in cell proliferation, cell death, inflammation, and DNA repair. The JNKs, which are encoded by three different Jnk loci, are now known to be regulated by many other stimuli, from pro-inflammatory cytokines to obesity, in addition to UV-irradiation. Targeted disruption of the Jnk loci in mice has proved to be a critical tool in better understanding their physiological functions. Such studies revealed that the JNKs play important roles in numerous cellular processes, including: programmed cell death, T cell differentiation, negative regulation of insulin signaling, control of fat deposition, and epithelial sheet migration. Importantly, the JNKs have become prime targets for drug development in several important clinical areas, including: inflammation, diabetes, and cancer. PMID- 16036613 TI - Understanding and modulating ageing. AB - Ageing is characterized by a progressive accumulation of molecular damage in nucleic acids, proteins and lipids. The inefficiency and failure of maintenance, repair and turnover pathways is the main cause of age-related accumulation of damage. Research in molecular gerontology is aimed at understanding the genetic and epigenetic regulation of survival and maintenance mechanisms at the levels of transcription, post-transcriptional processing, post-translational modifications, and interactions among various gene products. Concurrently, several approaches are being tried and tested to modulate ageing in a wide variety of organisms. The ultimate aim of such studies is to improve the quality of human life in old age and prolong the health-span. Various gerontomodulatory approaches include gene therapy, hormonal supplementation, nutritional modulation and intervention by free radical scavengers and other molecules. A recent approach is that of applying hormesis in ageing research and therapy, which is based on the principle of stimulation of maintenance and repair pathways by repeated exposure to mild stress. A combination of molecular, physiological and psychological modulatory approaches can realize "healthy ageing" as an achievable goal in the not-so distant future. PMID- 16036614 TI - How to clean the dirtiest place in the cell: cationic antioxidants as intramitochondrial ROS scavengers. AB - Membrane-penetrating triphenyl alkyl phosphonium cations have been suggested for many years in our group as having the ability to measure mitochondrial potential were recently used by Murphy as vehicles to specifically target CoQ to mitochondria. As was shown in our group, the phosphonium derivative of CoQ (MitoQ) easily penetrates a planar bilayer phospholipid membrane as a cation, generating 60 mV electric potential (Deltapsi) per a 10-fold MitoQ gradient. This means that MitoQ should be unequally distributed across the inner mitochondrial membrane, the intramitochondrial [MitoQ] = extramitochondrial [MitoQ] x 10(3) at 180 mV Deltapsi. In line with such a calculation, Murphy and his colleagues reported that antioxidant efficiency of MitoQ added to mitochondria or cells appears to be very much higher than of CoQ. It was found that H2O2-induced apoptosis (Murphy) and the H2O2-mediated bystander killing of the cultivated cells (our group) are completely arrested by pretreatement of the cells with 10( 10) - 10(-8) M MitoQ. These effects indicate that MitoQ and similar compounds may be promising in treatment of heart attack, stroke and other diseases accompanied by massive apoptosis in the injured tissue. The very fact that: (i) MitoQ is not only accumulated by mitochondria but also can be regenerated in its reduced form by mitochondrial respiratory chain, (ii) it is the mitochondrial interior that produces a large portion of reactive oxygen species (ROS) in our body, and (iii) the most sensitive ROS targets are localized in the mitochondrial matrix suggest the MitoQ-like compounds are promising tools of molecular therapy of aerobic cells. In line with this suggestion, we found that addition of MitoQ strongly improves structural and biochemical parameters of cultivated cells. As to cationic tetrapeptides, recently advertised as mitochondrially-targeted Deltapsi independent antioxidants, their effect is most probably mediated by an opioid activity inherent in some of these substances. PMID- 16036615 TI - Are genetically modified plants useful and safe? AB - So far, plants have been genetically modified essentially to achieve resistance to herbicides, or to pathogens (mainly insects, or viruses), but resistance to abiotic stresses (such as cold, heat, drought, or salt) is also being studied. Genetically modified (GM) plants with improved nutritional qualities have more recently been developed, such as plants containing higher proportions of unsaturated fatty acids (omega-3 and omega-6) in their oil (to prevent cardio vascular diseases), or containing beta-carotene as in the golden rice (to prevent vitamin A deficiency). Possible risks for human health (such as the production of allergenic proteins), or for the environment (such as the appearance of superweeds as a result from gene flow), should be carefully studied, and a science-based assessment of benefits vs. risks should be made on a case by case basis, both for GM plants and for plants obtained by conventional breeding methods. PMID- 16036616 TI - The ryanodine receptors Ca2+ release channels: cellular redox sensors? AB - The release of Ca2+ from intracellular stores mediated by ryanodine receptors (RyR) Ca2+ release channels is essential for striated muscle contraction and contributes to diverse neuronal functions including synaptic plasticity. Through Ca2+-induced Ca2+-release, RyR can amplify and propagate Ca2+ signals initially generated by Ca2+ entry into cardiac muscle cells or neurons. In contrast, RyR activation in skeletal muscle is under membrane potential control and does not require Ca2+ entry. Non-physiological or endogenous redox molecules can change RyR function via modification of a few RyR cysteine residues. This critical review will address the functional effects of RyR redox modification on Ca2+ release in skeletal muscle and cardiac muscle as well as in the activation of signaling cascades and transcriptional regulators required for synaptic plasticity in neurons. Specifically, the effects of endogenous redox-active agents, which induce S-nitrosylation or S-glutathionylation of particular channel cysteine residues, on the properties of muscle RyRs will be discussed. The effects of endogenous redox RyR modifications on cardiac preconditioning will be analyzed as well. In the hippocampus, sequential activation of ERKs and CREB is a requisite for Ca2+-dependent gene expression associated with long lasting synaptic plasticity. Results showing that reactive oxygen/nitrogen species modify RyR channels from neurons and the RyR-mediated sequential activation of neuronal ERKs and CREB produced by hydrogen peroxide and other stimuli will be also discussed. PMID- 16036617 TI - Control of DNA replication: regulation and activation of eukaryotic replicative helicase, MCM. AB - DNA replication is a key event of cell proliferation and the final target of signal transduction induced by growth factor stimulation. It is also strictly regulated during the ongoing cell cycle so that it occurs only once during S phase and that all the genetic materials are faithfully duplicated. DNA replication may be arrested or temporally inhibited due to a varieties of internal and external causes. Cells have developed intricate mechanisms to cope with the arrested replication forks to minimize the adversary effect on the stable maintenance of genetic materials. Helicases play a central role in DNA replication. In eukaryotes, MCM (minichromosome maintenance) protein complex plays essential roles as a replicative helicase. MCM4-6-7 complex possesses intrinsic DNA helicase activity which translocates on single-stranded DNA form 3' to 5'. Mammalian MCM4-6-7 helicase and ATPase activities are specifically stimulated by the presence of thymine-rich single-stranded DNA sequences onto which it is loaded. The activation appears to depend on the thymine content of this single-strand, and sequences derived from human replication origins can serve as potent activators of the MCM helicase. MCM is a prime target of Cdc7 kinase, known to be essential for activation of replication origins. We will discuss how the MCM may be activated at the replication origins by template DNA, phosphorylation, and interaction with other replicative proteins, and will present a model of how activation of MCM helicase by specific sequences may contribute to selection of replication initiation sites in higher eukaryotes. PMID- 16036618 TI - The thermogenic activity of rat brown adipose tissue and rabbit white muscle Ca2+ ATPase. AB - The Ca2+-ATPase (SERCA) found in vesicles derived from the sarco/endoplasmic reticulum vesicles of rats brown adipose tissue and rabbit white muscle were identified by gel electrophoresis, Western blot, electron microscopy and immunolabeling with gold particles. In both tissues, the isoform found was SERCA 1. The Ca2+ affinity of the fat SERCA 1 was different from the muscle isoform. The degree of uncoupling is estimated measuring the ratio between Ca2+ transport and ATP cleaved. In brown fat vesicles the degree of uncoupling varied depending on the Ca2+ concentration of the medium. This was not observed in vesicles derived from muscle. At all Ca2+ concentrations tested, the uncoupling was not related to Ca2+ leakage from the membrane and was far more pronounced in fat than in muscle vesicle. When a Ca2+ gradient was formed across the vesicles membrane the heat released during ATP hydrolysis varied between 22 and 26 Kcal/mol in both fat and muscle vesicles but in the absence of a gradient the heat released was 17 Kcal/mol in fat and 12 Kcal/mol in muscle. The data reported indicate that the SERCA 1 of brown adipocytes is far more thermogenic than the white muscle SERCA 1, and suggest that, in addition to storing Ca2+ inside the endoplasmic reticulum, the SERCA 1 may represent a source of heat production contributing to the thermogenic function of brown adipose tissue. PMID- 16036619 TI - Lysosomal cysteine proteases: structural features and their role in apoptosis. AB - Among the variety of proteolytic enzymes enormous progress has been seen recently in our understanding of lysosomal cysteine proteases, also known as cysteine cathepsins. These enzymes play a crucial role in diverse biological processes in physiological and pathological states, including genetic diseases. In the present review, their properties and structural features that are important to an understanding of their biological function are presented. Special emphasis is given to the newly discovered role of lysosomal cathepsins in apoptotic pathways. PMID- 16036620 TI - Loop movement and catalysis in creatine kinase. AB - Recently the crystal structure of creatine kinase from Torpedocalifornica was determined to 2.1 A. The dimeric structure revealed two different forms in the unit cell: one monomer was bound to a substrate, MgADP, and the other monomer was bound to a transition-state analogue complex composed of MgADP, nitrate and creatine. The most striking difference between the structures is the movement of two loops (comprising residues 60-70 and residues 323-333) into the active site in the transition state structure. This loop movement effectively occludes the active site from solvent, and the loops appear to be locked into place by a salt bridge formed between His66 and Asp326. His66 is of particular interest as it is located within a PGHP motif conserved in all creatine kinases but not found in other guanidino kinases. We have carried out alanine-scanning mutagenesis of each of the residues in the PGHP motif and determined that only the His66 plays a significant role in the creatine kinase reaction. Although neither residue interacts directly with the substrate, the interaction His66 and Asp326 appears to be important in providing the precise alignment of substrates necessary for phosphoryl group transfer. Finally, it is clear that neither His66 nor Asp326 are responsible for the pKs observed in the pH-rate profile for HMCK. PMID- 16036621 TI - Species-specific regulation of the alpha-2(I) procollagen gene by proximal promoter elements. AB - Transcriptional regulation of the human alpha 2(I) procollagen proximal promoter involves the interaction of trans-acting factors at the inverted CCAAT box (G/CBE) located at position -80 and an adjacent GGAGGCCC-box at -70. Both these elements have previously been shown to be essential for activity of the human promoter. This study investigated nucleotide differences at three sites (-74, -72 and -71) between the human and mouse promoters that were sufficient to abolish trans-acting factor binding with the mouse sequence (GGAGACGT). Two distinct DNA protein interactions were detected on the human -107/+54 promoter fragment while a single interaction was observed at the equivalent mouse promoter. One of these factors is the CCAAT-binding factor (CBF) and it's binding was observed on both the human and mouse promoters. Although the GGAGGCCC DNA-binding element was not detected on the mouse promoter, GGAGGCC-binding proteins were present in mouse nuclear extracts as observed by their interaction with the human promoter. Functional analysis of the human and mouse -343/+54 and -107/+54 promoter regions revealed significant differences between species; the human constructs having higher activity than the mouse. The differences in promoter activity between species may in part be a result of the nucleotide differences in the GGAGGCCC box. Mutations in this region of the human -107/+54 promoter prevented DNA protein interaction and lowered promoter activity. These results support the hypothesis that the GGAGGCCC-box in the human alpha 2(1) procollagen promoter has a regulatory function and that there exists a species-specific difference in transcription factor binding and regulation of the gene. PMID- 16036622 TI - Is the fatty acid synthesis pathway a good target for anti-malarial therapy? PMID- 16036623 TI - How I became a biochemist. PMID- 16036624 TI - IUB and IUBMB. PMID- 16036625 TI - The Internet and the elderly. PMID- 16036626 TI - Psychological barriers to Internet usage among older adults in the UK. AB - The Internet is an important tool in assisting the older population to lead independent and social lives. However, the majority of Internet users are under 55. This study investigated the following psychological barriers to Internet use by older adults: perceived usefulness, perceived ease of use, Internet efficacy, perceived complexity of navigation and perceived complexity of terminology. Twenty-three subjects (aged 55 - 75 years) were interviewed in the UK. The influences of age, computer and Internet experience, and training on these psychological barriers were explored. The results showed that the majority of the older adults who had a positive perception of usefulness, ease of use, and efficacy of the Internet or e-mail, used the Internet or e-mail more often. In addition, it was found that computer or Internet experience increased perceptions of ease of use and efficacy of the Internet and reduced perceived complexity of navigation. There was no difference between the two age groups (55 - 65 and 66 - 75 years) in these psychological barriers. It was concluded that increased marketing of the Internet (aimed at the older user), more simple and uniformly designed Internet pages, more user-friendly online help and error message terminology, and increased provision of training for the older user would assist uptake of the World Wide Web. PMID- 16036627 TI - The digital divide: Internet and e-mail use by the elderly. AB - The elderly are the fastest growing population group. Along with their rise in numbers is their volume of use of health care services, in both number and intensity. Since many elderly live alone, can their independence be maintained such that health care costs can decrease? With the rise in technology utilization, specifically the Internet, can the elderly utilize the Internet for health care purposes? Survey methodologies conducted for this study showed that the elderly can learn to use computers and are looking for methods to stay connected and be informed. Further longitudinal research is needed to measure clinical outcomes. PMID- 16036628 TI - ECG and XML: an instance of a possible XML schema for the ECG telemonitoring. AB - Management of many types of chronic diseases relies heavily on patients' self monitoring of their disease conditions. In recent years, Internet-based home telemonitoring systems allowing transmission of patient data to a central database and offering immediate access to the data by the care providers have become available. The adoption of Extensible Mark-up Language (XML) as a W3C standard has generated considerable interest in the potential value of this language in health informatics. However, the telemonitoring systems often work with only one or a few types of medical devices. This is because different medical devices produce different types of data, and the existing telemonitoring systems are generally built around a proprietary data schema. In this paper, we describe a generic data schema for a telemonitoring system that is applicable to different types of medical devices and different diseases, and then we present an architecture for the exchange of clinical information as data, signals of telemonitoring and clinical reports in the XML standard, up-to-date information in each electronic patient record and integration in real time with the information collected during the telemonitoring activities in the XML schema, between all the structures involved in the healthcare process of the patient. PMID- 16036629 TI - Identification of echocardiographic indices for the early detection of left ventricular systolic dysfunction in beta-thalassaemia via Self-Organizing Maps: a data-exploration study. AB - Congestive heart failure (CHF) remains the primary cause of death in patients suffering from beta-thalassaemia major. Its early detection allows the prompt initiation of aggressive chelation therapy, when the condition can still be reversed. We aimed at identifying echocardiographic indices for the early detection of left ventricular (LV) systolic dysfunction, the physiological abnormality underlying CHF, in these patients. We used Self-Organizing Maps (SOMs)--an artificial neural network--for identifying novel correlations within our Electronic Healthcare Record (EHCR) database on beta-thalassaemia. We sought echocardiographic parameters that are correlated to future deterioration of the LV ejection fraction and therefore constitute early signs of LV systolic dysfunction. At the same time, we evaluated SOMs as tools for exploring clinical datasets and make recommendations on the setup of the SOM algorithm that is appropriate for such tasks. We found that high values of the LV end-systolic diameter index and of the E/A ratio are early indications of LV systolic dysfunction. From a technical point of view, zero-mean unit-variance normalization of the input data, a large initial neighbourhood radius and a rectangular SOM grid produced optimal maps for the purpose of detecting clinical correlations. We have successfully used SOMs for exploring a clinical dataset and for creating novel medical hypotheses. A clinical study has been launched to confirm these hypotheses, and initial results are encouraging. PMID- 16036630 TI - Attitudes and skills of Hong Kong Chinese medicine practitioners towards computerization in practice: a cluster analysis. AB - The objective was to determine whether definable subtypes exist within a cohort of Hong Kong Chinese medicine practitioners (CMPs) as related to computer use in their clinical practices based on their skills, attitudes, and background factors. Data were collected using a structured questionnaire. The sample of 954 CMPs was recruited in Hong Kong. The study employs personal and demographic variables, attitudes, and a skills scale. A cluster analysis yielded three clusters. Each cluster represents a different profile of Hong Kong CMPs on computer use in their clinical practices. A clear profile of Hong Kong CMPs may benefit healthcare professionals in appropriate education or assistance to prompt the use of computers by CMPs, an officially recognized profession. PMID- 16036631 TI - A knowledge audit of the managers of primary care organizations: top priority is how to use routinely collected clinical data for quality improvement. AB - Technology has provided improved access to the rapidly expanding evidence base and to computerized clinical data recorded as part of routine care. A knowledge audit identifies from within this mass of information the knowledge requirements of a professional group or organization, enabling implementation of an appropriately tailored knowledge-management strategy. The objective of the study is to describe perceived knowledge gaps and recommend an appropriate knowledge management strategy for primary care. The sample comprised 18 senior managers of Primary Care Trusts: the Chairman, Chief Executive Officer, or Research and Development Lead. A series of interviews were recorded verbatim, transcribed and analysed. Knowledge requirements were broad, suggesting that a broadly based knowledge-management strategy is needed in primary care. The biggest gap in current knowledge identified is how to perform needs assessment and quality improvement using aggregated routinely collected, general practice computer data. PMID- 16036632 TI - A comparison of modulation of proliferation of thymocyte by testosterone, dehydroisoandrosterone and androstenedione in vitro. AB - Although the relationship between androgens and the immune system has been well established, the immunomodulatory function of different kinds of androgens on the cellular immunity remains to be defined. In the present study, the in vitro influences of testosterone, dehydroisoandrosterone and androstenedione on the Con A induced proliferation of thymocyte were determined. The results showed that both dehydroisoandrosterone and testosterone suppressed the Con A induced proliferation of thymocyte, while dehydroisoandrosterone was less potent than testosterone. In contrast, relatively high doses of androstenedione augmented the proliferative activities of thymocytes. Taken together, these data implied that small structural changes of androgens result in markedly different biological effects. These findings suggest that the balance between these steroids may be a significant determinant in maintaining immune homeostasis. PMID- 16036633 TI - Ureaplasma urealyticum infection related to seminal plasma immunosuppressive factors, semen pH and liquefaction duration. AB - In this study, semen samples from fertile and unexplained infertile men were explored for relationships between seminal plasma immunosuppressive factors (SPIFs), semen pH, liquefaction duration and infection of ureaplasma urealyticum (Uu). SPIFs activity was measured by way of counteracting complement. PH was detected by exact pH test paper. Liquefaction duration was observed at 37 degrees C. The results showed that Uu infection ratios of semen samples with abnormal SPIFs, pH or liquefaction duration were markedly higher than those of normal semen samples. It is suggested that Uu infection decreases the level of SPIFs, changes the pH in semen and prolongs the semen liquefaction so as to cause spermatic quality decline. The enhancement of SPIFs level may change the male body against Uu infection. PMID- 16036634 TI - Expression of beta-catenin in human testes with spermatogenic defects. AB - beta-catenin is a multifunctional molecule that functions in intercellular adhesion and signal transduction during assembly of AJs between Sertoli cells as well as between Sertoli cells and germ cells. To assess changes in the testicular beta-catenin in male infertility conditions, testicular tissues from obstructive azoospermia with normal spermatogenesis, spermatogenic arrest (SA) and Sertoli cell-only syndrome (SCO) patients were examined for immunohistochemical localization of beta-catenin. In normal spermatogenic tissue, expression of beta catenin was largely found in the Sertoli cell-germ cell (primarily spermatocytes) contact areas. Interestingly, perinuclear localization of beta-catenin was found in spermatocytes and spermatids. In spermatogenic arrest, beta-catenin in cell contact areas between Sertoli cells and germ cells was greatly decreased, but perinuclear beta-catenin in spermatocytes was not. In SCO, weak or negligible immunoreactivity of beta-catenin was found in cell contacts between Sertoli cells. Nuclear localization of beta-catenin was found in myotubular cells in all samples. Taken together, altered expression of beta-catenin in cell contacts within the seminiferous epithelia in spermatogenic arrest and SCO suggests that interactions between Sertoli cells and germ cell are crucial for expression of beta-catenin, and thus functional development of AJs in seminiferous epithelia in human testis. It should be also emphasized that perinuclear beta-catenin in germ cells may play a specific role in spermatogenesis. PMID- 16036635 TI - Power Doppler ultrasound mapping in nonobstructive azoospermic patients prior to testicular sperm extraction. AB - This study was conducted to localize the testicular regions, which have better blood circulation by power Doppler ultrasonography in patients with nonobstructive azoospermia before testicular sperm extraction (TESE), and to investigate whether these vascularized areas have a high sperm retrieval rate or not. We evaluated 110 testes of 55 cases that were diagnosed as nonobstructive azoospermia. The mean age of the study group was 33 years (range 26 to 42). Patients with Y chromosome microdeletions, karyotype and hormonal abnormalities (except elevated FSH levels) were excluded from the study. In all cases, testes were evaluated by power Doppler ultrasonography before testicular sperm extraction. Testis was divided vertically into five equal parts and the area with maximum vascularity was determined subjectively. During testicular sperm extraction, starting from best-perfused areas, biopsies were done. If no motile or sufficient amount of sperm was found, TESE procedure was tried on the contralateral testis. TESE were performed from 82 testes and for the regions that show good and poor vascularity. The sperm finding rate was 38% and 14%, respectively (OR = 3.55)(p = 0.001). Power Doppler ultrasound mapping of the testis in nonobstructive azoospermic cases is a reliable and informative method to assess spermatogenic foci. It is a noninvasive technique that minimizes the unnecessary removal of hormone producing tissue and gives chance to end the TESE earlier than currently practiced procedures. PMID- 16036636 TI - Effects of Tremella mesenterica on steroidogenesis in MA-10 mouse Leydig tumor cells. AB - Tremella mesenterica (TM), a yellow jelly mushroom, has been traditionally used as food and crude medicine to improve several kinds of symptoms in Chinese society for a long time. Recent studies have illustrated that the fractions of fruiting bodies of TM exhibit a significant hypoglycemic activity in diabetic mouse models, which usually suffer from sexual dysfunction. In a previous study, we showed that TM reduced plasma testosterone production in normal rats without any positive effect in diabetic rats. It evolved a question of TM directly regulating Leydig cell steroidogenesis. In this study, MA-10 mouse Leydig tumor cells were treated with vehicle, different dosages of TM with or without human chorionic gonadotropin (hCG 50 ng/ml) to clarify the effects. Results showed that TM at different dosages (0.01-10 mg/ml) did not have any effect on MA-10 cell steroidogenesis (p > 0.05). In the presence of hCG, there was an inhibitory trend that TA suppressed MA-10 cell progesterone production at 3 hr treatment with a statistically significant difference by the 10 mg/ml TM (p < 0.05). In time course effect, TM alone did not have any effect on MA-10 cell steroidogenesis from at 1, 2, 3, 6 and 12 hr (p > 0.05). However, TM did reduce hCG-treated MA-10 cell progesterone production at 1, 2 and 3 hr (p < 0.05), respectively. To determine whether TM would have adverse effects on MA-10 cell steroidogenesis in the presence of hCG, MTT assay and recovery studies were conducted. MTT assay indicated that TM had no effect on surviving cells. In addition, with the removal of TM, and then the addition of hCG (2 and 4 hr), progesterone levels were restored within 4 hr. Taken together, present studies suggested that TM suppressed hCG-treated steroidogenesis in MA-10 cells without any toxicity effect. PMID- 16036637 TI - Malignant phyllodes tumor of the prostate. AB - This is a report of a 70-year-old man with malignant phyllodes tumor of prostate. The retropubic prostatectomy was done. The stroma of the tumor was cellular and composed of elongated cells with spindle shaped nuclei and fragmented bizarre giant cells. After recovery from surgery, prophylatic radiotherapy was given over 2 months. A case report of a patient treated at our medical center and a review of the literature was done. PMID- 16036638 TI - Effect of body weight on sperm concentration in normozoospermic males. AB - A total of 274 men (aged: 26 +/- 4.9 years) with normozoospermia were enrolled into this study. Their body mass index (BMI: kg/m2) varied between 17 and 39. According to BMI, the patients were divided into four groups: Group 1: 17-20, Group 2: 20.1-25, Group 3: 25.1-30 and Group 4: 30.1-39. Twenty-nine subjects were found in the first, 96 in the second, 91 in the third and 58 men in the fourth group. Sperm concentration was significantly lower in the obese group (29 x 10(6)/ml, p < 0.05) than in the group of BMI 17-20, 20-25 and 25-30. In advance, in the obese group, sperm count continuously decreased with aging. We conclude that obesity is associated with a lower sperm count in case of normozoospermia. PMID- 16036639 TI - Effects of exposure to new inhalational anesthetics on spermatogenesis and sperm morphology in rabbits. AB - The effects of chronic exposure to new inhalation anesthetics (sevoflurane and isoflurane) on spermatogenesis and sperm morphology were examined in 23 rabbits, randomly divided in 3 groups. Rabbits received 20 exposure hours (four hours/day x 5 days), as follows: group I: 2.3% (1.2 MAC) sevoflurane + 2L/min oxygen, group II: 1.3% (1.2 MAC) isoflurane + 2L/min oxygen, and group III (control): 2L/min oxygen. Semen was collected on the 12th, 19th, 26th, 33rd, and 41st days of exposure. Sperm concentration, motility and morphological changes were evaluated. On the 41st day, testicular biopsies were taken and observed with light microscopy. Sperm concentration and motility significantly decreased in the sevoflurane and the isoflurane groups, compared to control. There were no significant changes in the control group. It is concluded that chronic exposure to the new inhalational anesthetics had negative effects on spermatogenesis and sperm morphology. PMID- 16036640 TI - Revision of hypo-osmotic swelling (HOS) test with a proposition of classified grading system: its comparisons with four different types of human sperm separation technique. AB - In this study, we arbitrarily classified the morphological changes of sperm under hypo-osmotic condition and compared the results with four different sperm separation techniques. The morphology of classification ranged from a good swelling (SG 3 > SG 2 > SG 1), to non-swelling but reactive (SG 0-R), and completely non-swelling (SG 0). Thirty fresh semen from patients were divided into 4 groups and each processed by washing, swim up, 2-layer column and real time micro-separation system. The prevalent patterns of the swollen sperm in sequence after treatment was SG 0 > SG 1 > SG 3 > SG2 > SG 0-R in fresh semen, and the proportion of SG 0 was almost half of the total % HOS test results. However, SG 3 became the most common swollen form with a significant increase in number with any method of sperm preparation after treatment. The real time technique yielded the highest % rate of SG 3 type of swollen spermatozoa and was the richest in concentration (53+/-3.9%, P < 0.05) as compared with that of swim up (26+/-4.6%), 2-layer (20+/-4.4%), sperm washing (23+/-3.9%), and fresh semen (17+/- 2.8%), respectively. By contrast, 2-layer collected more number of SG0-R (5.3+/-1.2%, P < 0.05) pattern of spermatozoa when compared to real time and swim up. A positive correlation (r = 0.81, P < 0.002) was shown between the % total HOS spermatozoa and total motility of fresh semen but not with morphology. Collectively, an abnormally functional sperm may exist in almost half of a total sperm count from men. It would appear that a better potential fertilization capacity may reside in the more swollen sperm from HOS test than the less swollen sperm and that may be more related with the motility rather than the morphology. The HOS classification and grading system appears valuable in further evaluating sperm quality. PMID- 16036641 TI - Human papillomavirus hpv-16 DNA as an epitheliotropic virus that induces hyperproliferation in squamous penile tissue. AB - The prevalence of human papillomavirus HPV-16DNA sequences in 57 penile carcinoma biopsies was examined using the polymerase chain reaction (PCR) with type specific internal probes, employing HPV consensus primers from the L1 region. The cases comprised 39 typical squamous cell carcinoma and 18 specimens with different subtype. PCR products were analyzed and HPV-16DNA was detected in a high percentage of specimens. Thirty-eight biopsies were HPV-16DNA positive. This determination was correlated with cellular differentiation and growth pattern. Our data corroborates that squamous cell carcinoma was invariably associated with HPV-16DNA. PMID- 16036642 TI - Respect as an organizing normative category for research ethics. PMID- 16036645 TI - Does uncle Sam really want you? A response to "Rethinking research Ethics" by Rosamond Rhodes (AJOB 5:1). PMID- 16036646 TI - Response to Athula Sumathipala and Sisira Siribaddana, "Revisiting 'freely given informed consent' in relation to the developing world: the role of an Ombudsman" (AJOB 4:3). PMID- 16036647 TI - Response to Fritz Allhoff, "Telomeres and the ethics of human cloning" (AJOB 4:2). PMID- 16036648 TI - Lessons in conflict of interest: the construction of the martyrdom of David Healy and the dilemma of bioethics. AB - Bioethics journals have lagged behind medical and science journals in exploring the threat of conflict of interest (COI) to the integrity of publications. Some recent discussions of COI that have occurred in the bioethics literature are reviewed. Discussions of what has been termed the "Healy affair" unintentionally demonstrate that the direct and indirect influence of undisclosed COI may come from those who call for protection from the undue influence of industry. Paradoxically, the nature and tone of current discussions may serve to dull sensitivities to what is indeed a serious set of issues facing bioethics. Some proposals are presented to address COI and other challenges to the integrity of bioethics and its journals. COI is too important a topic to be left to ideologues, and there is no substitute for readers' caution and skepticism as tools in dealing with the full range of biases that exist in published papers. PMID- 16036649 TI - Reflections on 'Rethinking research ethics'. PMID- 16036650 TI - Consequentialism and harsh interrogations. AB - With this issue, we begin a regular feature on bioethics and public health. We welcome Matthew K. Wynia, M.D., M.P.H., Director of the Institute for Ethics of the American Medical Association as our new Contributing Editor. If you have comments or suggestions regarding this feature, please email us at manuscript@ bioethics.net. PMID- 16036651 TI - Rethinking research ethics. AB - Contemporary research ethics policies started with reflection on the atrocities perpetrated upon concentration camp inmates by Nazi doctors. Apparently, as a consequence of that experience, the policies that now guide human subject research focus on the protection of human subjects by making informed consent the centerpiece of regulatory attention. I take the choice of context for policy design, the initial prioritization of informed consent, and several associated conceptual missteps, to have set research ethics off in the wrong direction. The aim of this paper is to sort out these confusions and their implications and to offer instead a straightforward framework for considering the ethical conduct of human subject research. In the course of this discussion I clarify different senses of autonomy that have been confounded and present more intelligible justifications for informed consent. I also take issue with several of the now accepted dogmas that govern research ethics. These include: the primacy of informed consent, the protection of the vulnerable, the substitution of beneficence for research's social purpose, and the introduction of an untenable distinction between innovation and research. PMID- 16036652 TI - Some questionable premises about research ethics. PMID- 16036653 TI - How not to rethink research ethics. PMID- 16036654 TI - Does research ethics rest on a mistake? PMID- 16036655 TI - Waste not, want not: cognitive impairment should not preclude research participation. PMID- 16036656 TI - Does research ethics rest on a mistake? The common good, reasonable risk and social justice. PMID- 16036657 TI - Additional thoughts on rethinking research ethics. PMID- 16036658 TI - Research versus innovation: real differences. PMID- 16036659 TI - Beginning anew: same principles, different direction for research ethics. PMID- 16036660 TI - Context is key for voluntary and informed consent. PMID- 16036661 TI - Research participation: are we subject to a duty? PMID- 16036662 TI - Free-riding and research ethics. PMID- 16036663 TI - Putting the "ethics" into "research ethics". PMID- 16036664 TI - Histories of mistrust and protectionism: disadvantaged minority groups and human subject research policies. PMID- 16036665 TI - Trust, understanding and utopia in the research setting. PMID- 16036666 TI - Pediatric do-not-attempt-resuscitation orders and public schools: a national assessment of policies and laws. AB - Some children living with life-shortening medical conditions may wish to attend school without the threat of having resuscitation attempted in the event of cardiopulmonary arrest on the school premises. Despite recent attention to in school do-not-attempt-resuscitation (DNAR) orders, no assessment of state laws or school policies has yet been made. We therefore sought to survey a national sample of prominent school districts and situate their policies in the context of relevant state laws. Most (80%) school districts sampled did not have policies, regulations, or protocols for dealing with student DNARs. A similar majority (76%) either would not honor student DNARs or were uncertain about whether they could. Frequent contradictions between school policies and state laws also exist. Consequently, children living with life-shortening conditions who have DNARs may not have these orders honored if cardiopulmonary arrest were to occur on school premises. Coordinated efforts are needed to harmonize school district, state, and federal approaches in order to support children and families' right to have important medical decisions honored. PMID- 16036667 TI - School DNAR in the real world. PMID- 16036668 TI - Back to the future: overcoming reluctance to honor in-school DNAR orders. PMID- 16036669 TI - Going to school to die: equal treatment for well and ill children. PMID- 16036670 TI - The challenge of DNAR orders in schools. PMID- 16036671 TI - DNAR in schools: questions and concerns. PMID- 16036672 TI - Legal frameworks for addressing the well-being of terminally ill children. PMID- 16036673 TI - DNAR in the schools: watch your language! PMID- 16036674 TI - Ensuring that difficult decisions are honored--even in school settings. PMID- 16036675 TI - The spectrum of our obligations: DNR in public schools. PMID- 16036676 TI - Nurses at the helm: implementing DNAR orders in the public school setting. PMID- 16036677 TI - The first step: DNAR outside the hospital and the role of pediatric medical care providers. PMID- 16036678 TI - Pediatric resuscitation: questioning DNAR legitimacy and offering an alternative decision-making model. AB - The views expressed in this paper are those of the author and do not necessarily represent those of the American Medical Association. PMID- 16036682 TI - Dying for food. PMID- 16036683 TI - Slouching toward policy: lazy bioethics and the perils of science fiction. PMID- 16036687 TI - Science, faith and AIDS: the battle over harm reduction. PMID- 16036689 TI - Neuroimaging: revolutionary research tool or a post-modern phrenology? PMID- 16036688 TI - Imaging or imagining? A neuroethics challenge informed by genetics. AB - From a twenty-first century partnership between bioethics and neuroscience, the modern field of neuroethics is emerging, and technologies enabling functional neuroimaging with unprecedented sensitivity have brought new ethical, social and legal issues to the forefront. Some issues, akin to those surrounding modern genetics, raise critical questions regarding prediction of disease, privacy and identity. However, with new and still-evolving insights into our neurobiology and previously unquantifiable features of profoundly personal behaviors such as social attitude, value and moral agency, the difficulty of carefully and properly interpreting the relationship between brain findings and our own self-concept is unprecedented. Therefore, while the ethics of genetics provides a legitimate starting point--even a backbone--for tackling ethical issues in neuroimaging, they do not suffice. Drawing on recent neuroimaging findings and their plausible real-world applications, we argue that interpretation of neuroimaging data is a key epistemological and ethical challenge. This challenge is two-fold. First, at the scientific level, the sheer complexity of neuroscience research poses challenges for integration of knowledge and meaningful interpretation of data. Second, at the social and cultural level, we find that interpretations of imaging studies are bound by cultural and anthropological frameworks. In particular, the introduction of concepts of self and personhood in neuroimaging illustrates the interaction of interpretation levels and is a major reason why ethical reflection on genetics will only partially help settle neuroethical issues. Indeed, ethical interpretation of such findings will necessitate not only traditional bioethical input but also a wider perspective on the construction of scientific knowledge. PMID- 16036690 TI - On the proliferation of bioethics sub-disciplines: do we really need "genethics" and "neuroethics"? PMID- 16036691 TI - Brains, genes, and the making of the self. PMID- 16036692 TI - Caution in leaping from functional imaging to functional neurosurgery. PMID- 16036693 TI - Framing neuroethics: a sociological assessment of the neuroethical imagination. PMID- 16036694 TI - Finding the right compass for issue-mapping in neuroimaging. PMID- 16036695 TI - Imagining a neuroethics which would go further than genethics. PMID- 16036696 TI - Neuroethics: a philosophical challenge. PMID- 16036697 TI - Neuroethics, new ethics? PMID- 16036698 TI - Neuroscience and metaphysics. PMID- 16036699 TI - Neuro exceptionalism? PMID- 16036700 TI - Emerging neurotechnologies for lie-detection: promises and perils. AB - Detection of deception and confirmation of truth telling with conventional polygraphy raised a host of technical and ethical issues. Recently, newer methods of recording electromagnetic signals from the brain show promise in permitting the detection of deception or truth telling. Some are even being promoted as more accurate than conventional polygraphy. While the new technologies raise issues of personal privacy, acceptable forensic application, and other social issues, the focus of this paper is the technical limitations of the developing technology. Those limitations include the measurement validity of the new technologies, which remains largely unknown. Another set of questions pertains to the psychological paradigms used to model or constrain the target behavior. Finally, there is little standardization in the field, and the vulnerability of the techniques to countermeasures is unknown. Premature application of these technologies outside of research settings should be resisted, and the social conversation about the appropriate parameters of its civil, forensic, and security use should begin. PMID- 16036701 TI - Premarket approval regulation for lie detections: an idea whose time may be coming. PMID- 16036702 TI - Dual use and the "moral taint" problem. PMID- 16036703 TI - Spy versus spy. PMID- 16036704 TI - The brain doesn't lie. PMID- 16036705 TI - The Orwellian threat to emerging neurodiagnostic technologies. PMID- 16036706 TI - Can we scan for truth in a society of liars? PMID- 16036707 TI - Keeping an open mind: what legal safeguards are needed? PMID- 16036708 TI - Searching the brain: the Fourth Amendment implications of brain-based deception detection devices. PMID- 16036714 TI - Development of controlled release spheroids using natural polysaccharide as release modifier. AB - A polysaccharide hydrogel was isolated from the seeds of Tamarindus indica (tamarind) and was used as release modifier for the preparation of diclofenac sodium spheroids, using extrusion-spheronization technique. The process was studied for the effect of variables to arrive at spheroids with satisfactory particle shape, size and size-distribution. The prepared spheroids were characterized for surface morphology, qualitative surface porosity, friability, bulk density, and flow properties. The in vitro release studies exhibited a zero order release kinetics that was confirmed by Higuchi's and Peppas' models. A credible correlation was obtained among swelling index, viscosity, surface roughness of the polysaccharide, and in vitro dissolution profile of the spheroids. In the comparative bioavailability study, we found that the developed spheroids were able to sustain the drug release over 8 hr and could improve the extent of absorption and bioavailability of the drug. PMID- 16036715 TI - Solid lipid nanoparticles bearing flurbiprofen for transdermal delivery. AB - Topical application of the drugs at the pathological sites offer potential advantages of delivering the drug directly to the site of action and thus producing high tissue concentrations of the drug. The solid lipid nanoparticles (SLN) bearing flurbiprofen were prepared by microemulsion method by dispersing o/w microemulsion in a cold aqueous surfactant medium under mechanical stirring. The SLN gel was prepared by adding SLN dispersion to polyacrylamide gel prepared by using polyacrylamide (0.5%), glycerol (10%), and water (69.5%). Shape and surface morphology was determined by scanning electron microscopy that revealed fairly spherical shape of the formulation. Percent drug entrapment was higher in SLN dispersion in comparison to SLN gel formulations. In vitro drug release, determined using cellophane membrane, showed that SLN dispersion exhibited higher drug release compared with SLN gel formulations. Both the SLN dispersion and SLN gel formulation possessed a sustained drug release over a 24-hr period, but this sustained effect was more pronounced with SLN-gel formulations. The percent inhibition of edema after 8 hr was 55.51 +/- 0.26% in case of SLN-T4-gel, whereas flurbiprofen and SLN-T4 dispersion exhibited 28.81 +/- 0.46 and 31.89 +/- 0.82 inhibition of edema. The SLN-T4-gel not only decreased the inflammation to larger magnitude, but also sustained its effect. PMID- 16036716 TI - Comparison of two pegylated copolymeric micelles and their potential as drug carriers. AB - The aim of this study was to evaluate the ability of forming micelles from two types of synthesized diblock pegylated amphiphilic copolymers and their potential as a drug carrier. Two lactone monomers, epsilon-caprolactone (CL) and delta valerolactone (VL), were copolymerized with methoxy poly(ethylene glycol) (MePEG), respectively. The properties of copolymers were investigated and their biocompatibility was tested through an in vitro cytotoxicity study. The influences of the type of lactone monomer (CL and VL) and the feed molar ratios of lactone/MePEG (50/1, 80/1, 160/1) on the performance and release behavior of drug-loaded micelles were investigated. The opening of CL and VL rings by MePEG was efficient, and the pegylation of poly(lactone)s allowed copolymers possessing amphiphilic property and efficiently self-assembled to form micelles with a low critical micelle concentration (CMC) in the range of 10(-7)-10(-8) M. The nano sized micelles were able to incorporate hydrophobic drug and regulate drug release, and the release of drug was dominated by the hydrophobic poly(lactone) chain length. Although both amphiphilic copolymers exhibited similar controlled release character, the PCL/MePEG micelles possessed lower CMC, higher biocompatibility, and higher drug loading than PVL/MePEG micelles. These suggested that results choosing pegylated PCL as a drug carrier could be better than PVL/MePEG. PMID- 16036717 TI - Radical cross-linked albumin microspheres as potential drug delivery systems: preparation and in vitro studies. AB - The aim of this research is the preparation of acryloylated bovine serum albumin microspheres and the evaluation of their employment in drug delivery. The influence of preparation parameters on albumin microspheres and the chemicophysical properties of loaded drugs were investigated. In particular, we focused our attention on acylation albumin degree, amount of acryloylated albumin against comonomer in the polymerization step, and finally the release profile. We considered on the interaction drug-matrix, the fuctionalization degree of albumin, and the water affinity of matrix. PMID- 16036722 TI - Long-term effectiveness of spaced-retrieval memory training for older adults with probable Alzheimer's disease. AB - Ten older adults with probable Alzheimer's disease (AD) were trained to recall everyday objects using the spaced-retrieval technique. Five persons had participated in a prior spaced-retrieval program (Cherry & Simmons-D'Gerolamo, 1999, Clinical Gerontologist, 20, 39-63). The authors retested these original participants to measure the long-term effectiveness of spaced-retrieval training. Five new participants were included for comparison purposes. During training, participants selected a target object from an array of items at increasingly longer retention intervals. To enhance the effectiveness of spaced-retrieval training, two original and two new participants were given a target object orientation task prior to training. Positive effects of prior experience occurred for most of the original participants. The orientation task enhanced memory for the target objects. Implications for memory remediation in late adulthood are considered. PMID- 16036721 TI - How feelings of stereotype threat influence older adults' memory performance. AB - The purpose of the present research was to explore the role of stereotype threat as a mediator of older people's memory performance under different instructional sets. In three studies, younger and older participants completed a memory test that was either framed as a memorization or as an impression formation task. Across these studies, memory performance was greater for younger than for older adults and was higher in the impression formation than memorization condition, but was not different for older adults in the two instruction conditions. These results also showed that age differences in memory performance were mediated by participants' feelings of stereotype threat, such that age was positively related to stereotype threat and stereotype threat was negatively related to memory performance. These data demonstrate that concerns about being negatively stereotyped influence age differences in memory performance, and that the effects of these feelings on performance are not easily reduced by reframing the task instructions. PMID- 16036723 TI - A growth curve model of learning acquisition among cognitively normal older adults. AB - The objective of this study was to model recall and learning on the Auditory Verbal Learning Test using latent growth curve techniques. Participants were older adults recruited for the ACTIVE cognitive intervention pilot. A series of nested models revealed that an approximately logarithmic growth curve model provided optimal fit to the data. Although recall and learning factors were statistically uncorrelated, a fitted multivariate model suggested that initial recall was significantly associated with demographic characteristics but unrelated to health factors and cognitive abilities. Individual differences in learning were related to race/ethnicity, speed of processing, verbal knowledge, and global cognitive function level. These results suggest that failing to recognize initial recall and learning as distinct constructs clouds the interpretation of supraspan memory tasks. PMID- 16036724 TI - Listening to spoken text: adult age differences as revealed by self-paced listening. AB - An experiment is reported in which young and older adults heard spoken narratives presented in a segment-by-segment fashion using the auditory moving window (AMW) technique. Participants were instructed to initiate the presentation of each segment at their own pace, their goal being to insure good comprehension of the main ideas of the narratives. The pattern of pause times across passages was compared for passages being heard for the first time (novel condition) or after the participants had heard the passages several times before (familiar condition). The analysis of participants' pause durations in pacing through the passages suggests that although young and older adults respond similarly at the textbase level of processing, older adults do not allocate additional resources at the start of a passage in order to develop a mental model of the narrative. This pattern differs from that typically found in reading time studies. PMID- 16036725 TI - When familiar social partners are selected in open-ended situations: further tests of the socioemotional selectivity theory. AB - Socioemotional selectivity theory (SST; Carstensen, 1995, Current Directions in Psychological Science, 4, 151-156) predicts that novel social partners are preferred in open-ended situations, whereas familiar social partners are preferred in future-limited situations. The authors attempted to generalize past research to new familiar and novel partner options. Studies 1 (N=144; undergraduates, community-dwelling adults ages 65 to 95) and 2 (N=336 community dwelling participants ages 11 to 89) indicated that young and older participants in a future-limited situation preferred familiar partners. However, with different social partner options than have been used in previous research, young participants in an open-ended situation also preferred a familiar partner, contrary to the predictions of SST. PMID- 16036726 TI - Medial temporal lobe atrophy, apolipoprotein genotype, and plasma homocysteine in Sri Lankan patients with Alzheimer's disease. AB - The authors studied the association of Alzheimer's disease (AD) with total plasma homocysteine (tHcy) and apolipoprotein E (apoE) genotype, and the usefulness of measuring medial temporal lobe thickness (MTL) thickness for the diagnosis of AD in Sri Lankan patients. Using criteria of the NINCDS-ADRDA, 23 AD patients and 21 controls were recruited. All underwent MTL-oriented computed tomographic (CT) scans, measurement of plasma tHcy, and apoE genotyping. Mean plasma tHcy was significantly higher in AD patients than controls (p=.001). This association was independent of age, sex, body mass index (BMI), serum folate and vitamin B12, and serum creatinine. The frequency of apoE4 allele was significantly higher (p=.003) in AD patients, and the adjusted odds ratio of AD for the presence of one or more apoE4 alleles compared with none was 10.39 (95% CI 1.77-61.10; p=.010). The mean minimum MTL thickness was significantly higher in control subjects compared to that of AD patients (p<.001). This first report of apoE4, plasma tHcy, MTL thickness, and AD from Sri Lanka shows that high plasma tHcy, the presence of apoE4 allele, and MTL atrophy are associated with AD. PMID- 16036727 TI - Beyond the 2004 tsunami: health diplomacy as a response. PMID- 16036728 TI - Cancer information needs in rural areas. AB - Although cancer presents obstacles for all who experience it, persons in rural communities must negotiate additional challenges. This study determined the cancer information (CI) needs and the CI-seeking behavior and preferences among rural-dwelling persons. Patients (N = 801) = 50 years of age seen in 36 rural Kansas primary care practices completed a Cancer Care Information Needs Survey (CCINS); physicians completed a cancer resource knowledge and preference survey. Of the 801 patients, 184 (23%) reported a CI need. Of these 184 patients, 45% reported either not discussing cancer or having insufficient discussion time with their physicians; 44% needed more information after consulting their physician. Patients more likely to report a CI need were young, female, Internet users, persons with a prior cancer diagnosis, and persons seeing male physicians or physicians in group/multispecialty practices. Patients and physicians were unfamiliar with services provided by national cancer organizations. Physicians are a primary CI source; however, patients who need CI report insufficient cancer discussion time with their physician and need more CI after consulting their physician. Promoting access to national CI sources could bridge the CI needs gap that exists in rural areas currently. PMID- 16036729 TI - Health-related message boards/chat rooms on the Web: discussion content and implications for pharmaceutical sponsorships. AB - This content analysis study examined health-related message boards to better understand who is using this on-line health information and support device and what topics they are discussing. Besides needing to understand this support and coping mechanism for individuals, this has become an increasingly important topic for health communicators to understand because the Health and Human Services' (HHS) Inspector General recently gave permission to a pharmaceutical manufacturer for sponsorship of a disease management chat room. Very little research has been done on the content of these message boards/chat rooms. Key findings include that the most commonly discussed medical topics were medical treatments and drugs (often specific brands) and that these boards are clearly important sources of information and emotional support. Implication and future research are discussed. PMID- 16036730 TI - The effectiveness of media use in health education: evaluation of an HIV/AIDS radio [corrected] campaign in Ethiopia. AB - An effective vehicle to change behaviors is entertainment education. To demonstrate entertainment education effects, researchers must first indicate that participants have been exposed to their program. Exposure to effective programs has been associated with increases in knowledge about program topics, attitude change, and self-efficacious perceptions. The purpose of this study was to develop and test a new exposure technique that accurately and precisely determines direct exposure levels to Ethiopia's Journey of Life. Overall, the study found very high listenership, storyline recall, liking of the program, and strong desire to change behavior while maintaining low error rates in terms of verbatim recall of storylines and reported listenership to a fictitious program. PMID- 16036731 TI - Examining college students' intentions to become organ donors. AB - 502 university students completed survey items on attitudes, experiences, knowledge, and behaviors related to organ and tissue donation (OTD). Despite positive attitudes toward organ donation, only 11% of students formally have declared their intentions to donate through the state registry or by signing an organ card. When asked to report why they have not signed an organ donor card/registry, students reported, "not considering the topic," "intentions to donate in the future," and "general negative attitudes" among other reasons. Students also reported a generally positive attitude toward the topic of OTD and moderate to strong intentions to become organ donors in the future despite feeling somewhat uninformed on the topic. The results are discussed in relation to future campaign message strategies to promote OTD to university students. PMID- 16036732 TI - Embedding health messages into entertainment television: effect on gay men's response to a syphilis outbreak. AB - The entertainment education (EE) approach seeks to impact audiences' health behavior by embedding messages in mass media productions, storylines, and characters that appeal strongly to them. Effect on behavioral intentions was examined following a storyline about syphilis in men who have sex with men (MSM) presented in a popular dramatic series. Five hundred and one MSM drawn from gay Internet chat rooms completed the questionnaire. Differences in item responses between those who did and did not view the syphilis storyline were calculated, and linear regression was used to examine predictors of intentions to take action. Those who viewed the syphilis storyline were more likely to report intention to get screened and to tell others to get screened for syphilis. Seeing the episode was a predictor of these intentions. Education was also a predictor of intention to tell others to get screened. Results suggest that exposure to a storyline about syphilis in gay men had a positive public health outcome on users of Internet chat rooms for MSM. Further studies are warranted to examine the extent to which and how the EE approach can produce health-relevant outcomes for U.S. populations, just as the approach has done in the developing world. PMID- 16036733 TI - Understanding the role of cigarette promotion and youth smoking in a changing marketing environment. AB - In 2001, $11.21 billion was spent on domestic cigarette advertising and promotion, an increase of 16.9% over 2000. This article explains how cigarette industry efforts stimulate demand and encourage smoking within the context of recent changes, including the 1998 Master Settlement Agreement (MSA) and resulting litigation, and variations in tobacco marketing policies. Communication concepts are combined with adolescent development concepts to explain how youth are impacted. Industry documents and current syndicated research data are used to reveal and explain key concepts. PMID- 16036736 TI - Writing with Ev--words to transform science into action. PMID- 16036737 TI - Commentaries: Ev Roger's gifts. PMID- 16036738 TI - The association between knowledge and attitudes about genetic testing for cancer risk in the United States. AB - Attitudes about genetic testing are likely to be an important determinant of uptake of predictive genetic tests among the general public. Several prior studies have suggested that positive attitudes about genetic testing may be inversely related to knowledge about genetic testing. We conducted a random-digit dialing (RDD) telephone survey of 961 adults in the continental United States to determine the associations among knowledge of, attitudes about, and perceptions of eligibility for genetic testing for cancer risk. Knowledge about genetic testing for cancer risk was generally high, with a mean accuracy score of 72%. Attitudes about genetic testing for cancer risk were also generally positive, with 87% of respondents reporting genetic testing for cancer risk would be used to help doctors manage their health care and 85% to help scientists find cures for diseases. In contrast, 58% of respondents thought genetic testing for cancer risk would be used to prevent them from getting health insurance and 31% to allow the government to label groups as inferior. Twenty-nine percent of respondents thought they were currently eligible for testing. After adjustment for sociodemographic characteristics and family cancer history, higher knowledge was correlated with more positive attitudes about testing, but not with negative attitudes or perceptions of testing eligibility. Family history was positively associated with perceptions of eligibility (OR 3.49, 95% CI 2.36-5.18), and higher levels of education were inversely associated with perceptions of eligibility (OR 0.55, 95% CI 0.32-0.94 for comparison of college or higher vs. less than high school). These results suggest that most members of the general public are knowledgeable and have positive attitudes about genetic testing for cancer risk and that greater knowledge is correlated with more positive attitudes about the benefits of testing. PMID- 16036739 TI - Genomics--the perfect information-seeking research problem. AB - The intersection of the genetics era and information age poses unique and daunting challenges for health consumers who may not have the health literacy to keep pace. While rapid advances in genetics research promise enhanced care, the inherent complexities and individualistic nature of genetic information have resulted in a challenging information environment. The technical possibilities for acquiring genomic information are increasing at an exponential pace, as are the scientific advances relating to it. Furthermore, societal reactions to genomics, and possible privacy and discrimination issues, may constitute significant constraints. The health care infrastructure also has its limits, given the severe shortage of qualified cancer genetic counselors and general practitioners who are unprepared to address genetics, creating a demand for creative approaches to service delivery. The combination of individual salience, low health literacy, the consumer movement, and important policy problems, then makes genomics the perfect information seeking research problem. PMID- 16036740 TI - Messages from teens on the big screen: smoking, drinking, and drug use in teen centered films. AB - Smoking, drinking, and drug use endure as popular yet dangerous behaviors among American teenagers. Films have been cited as potential influences on teens' attitudes toward and initiation of substance use. Social cognitive theory suggests that teen viewers may be especially likely to learn from teen models who they perceive as similar, desirable, and attractive. Yet, to date, no studies systematically have analyzed teen characters in films to assess the frequency, nature, and experienced consequences of substance use depictions. Assessments of content are necessary precursors to effects studies because they can identify patterns of representations that warrant further examination. Accordingly, a content analysis of top grossing films from 1999, 2000, and 2001 was conducted. Overall, two-fifths of teen characters drank alcohol, one-sixth smoked cigarettes, and one-seventh used illicit drugs (N=146). Almost no differences existed between substance users and nonusers with regard to physical attractiveness, socioeconomic status (SES), virtuosity, or gender. Drinkers and drug users were unlikely to suffer any consequences--let alone negative consequences--in either the short or long term. Characters rarely were shown refusing offers to drink or do drugs, or regretting their substance usage. Girls were more likely than boys to be shown engaging in multiple substance use activities (e.g., smoking and drinking). Overall, recent teen-centered films may teach teen viewers that substance use is relatively common, mostly risk-free, and appropriate for anyone. PMID- 16036741 TI - HIV/AIDS stories on the World Wide Web and transformation perspective. AB - Survivor stories have been an important part of therapy and social support for persons living with human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome AIDS (PLWHAs), and the World Wide Web has made these stories accessible to a vast audience. These stories are examined in the light of the concept of "transformation perspective" defined as a self-communicative experience that changes an individual's life so that priorities and self-identity are refocused. The trigger event that alters the individual's life might be a diagnosis with cancer, HIV, diabetes, or some other serious illness; divorce; financial tragedy; unemployment; or retirement. The "disorienting dilemma," according to Mezirow, leads to self-examination, and thus to changes in the individual's frame of reference. The present study found that PLWHA's stories with more details were more likely to reflect transformation perspective. PMID- 16036742 TI - Women Connect! Strengthening communications to meet sexual and reproductive health challenges. AB - Women's nongovernmental organizations (NGOs) have significant comparative advantage for addressing sexual and reproductive health challenges facing women and families. This article describes an initiative to assist women's NGOs in developing greater skills using media and information communication technology for communicating women's health messages. Participating women's groups in Africa undertook innovative media projects--radio broadcasts on human immunodeficiency virus (HIV) and family planning, an antiviolence campaign, media campaigns on avoiding teen pregnancy--and designed websites, established Internet cafes, and downloaded health information from the Internet. Lessons learned offer guidance for collaboration with women's NGOs everywhere to strengthen communication for addressing critical sexual and reproductive health issues. PMID- 16036744 TI - Exposures of elderly volunteers with and without chronic obstructive pulmonary disease (COPD) to concentrated ambient fine particulate pollution. AB - The elderly and individuals who have chronic obstructive pulmonary disease (COPD) may be sensitive to particulate matter (PM) air pollution. We evaluated short term health responses of 13 elderly volunteers with COPD and 6 age-matched healthy adults to controlled exposures of ambient PM pollution in suburban Los Angeles. Using a Harvard particle concentrator and a whole-body chamber, we exposed each person on separate occasions to approximately 200 microg/m(3) concentrated ambient particles (CAP) less than 2.5 mum in diameter and to filtered air (FA). Each exposure lasted 2 h with intermittent mild exercise. We found no significant effects of CAP on symptoms, spirometry, or induced sputum. A significant negative effect of CAP on arterial oxygenation (measured by pulse oximetry) immediately postexposure was more pronounced in healthy subjects. Peripheral blood basophils increased after CAP in healthy but not in COPD subjects. In both groups, red cell counts increased slightly 1 day after exposure to FA but not to CAP. Preexposure ectopic heartbeats were infrequent in healthy subjects, but increased modestly during/after CAP exposure relative to FA. Ectopic beats were more frequent in COPD subjects, but decreased modestly during/after CAP relative to FA. Heart-rate variability over multi-hour intervals was lower after CAP than after FA in healthy elderly subjects but not in COPD subjects. Thus, in this initial small-scale study of older volunteers experimentally exposed to ambient PM, some acute cardiopulmonary responses were consistent with effects reported from epidemiologic studies. Unexpectedly, individuals with COPD appeared less susceptible than healthy elderly individuals. Further investigation of older adults is warranted to understand the pathophysiology and public health significance of these findings. PMID- 16036745 TI - The biopersistence of brazilian chrysotile asbestos following inhalation. AB - With the initial understanding of the relationship of asbestos to disease, little information was available on whether the two different groups of minerals that are called asbestos were of similar or different potency in causing disease. Asbestos was often described as a durable fiber that if inhaled would remain in the lung and cause disease. It has been only more recently, with the development of a standardized protocol for evaluating the biopersistence of mineral fibers in the lung, that the clearance kinetics of the serpentine chrysotile have been shown to be dramatically different from those of amphibole asbestos, with chrysotile clearing rapidly from the lung. In addition, recent epidemiology studies also differentiate chrysotile from amphibole asbestos. The biopersistence studies mentioned have indicated that chrysotile from Canada and California clear rapidly from the lung once inhaled. However, variations in chrysotile mineralogy have been reported depending upon the region. This is most likely associated with variations in the forces which created the chrysotile fibers centuries ago. In the present study, the dynamics and rate of clearance of chrysotile from the Cana Brava mine in central Brazil was evaluated in a comparable inhalation biopersistence study in the rat. For synthetic vitreous fibers, the biopersistence of the fibers longer than 20 microm has been found to be directly related to their potential to cause disease. This study was designed to determine lung clearance (biopersistence) and translocation and distribution within the lung. As the long fibers have been shown to have the greatest potential for pathogenicity, the chrysotile samples were specifically chosen to have more than 450 fibers/cm(3) longer than 20 microm in length present in the exposure aerosol. For the fiber clearance study (lung digestions), at 1 day, 2 days, 7 days, 2 wk, 1 mo, 3 mo, 6 mo, and 12 mo following a 5-day (6 h/day) inhalation exposure, the lungs from groups of animals were digested by low-temperature plasma ashing and subsequently analyzed by transmission electron microscopy (at the GSA Corp.) for total chrysotile fiber number in the lungs and chrysotile fiber size (length and diameter) distribution in the lungs. This lung digestion procedure digests the entire lung with no possibility of identifying where in the lung the fibers are located. A fiber distribution study (with confocal microscopy) was included in order to identify where in the lung the fibers were located. At 2 days, 2 wk, 3 mo, 6 mo, and 12 mo postexposure, the lungs from groups of animals were analyzed by confocal microscopy to determine the anatomic fate, orientation, and distribution of the retained chrysotile fibrils deposited on airways and those fibers translocated to the broncho-associated lymphoid tissue (BALT) subjacent to bronchioles in rat lungs. While the translocation of fibers to the BALT and lymphatic tissue is considered important as in cases of human's with asbestos related disease, there has been no report in the literature of pathological changes in the BALT and lymphatic tissue stemming from asbestos. Thus, if the fibers are removed to these tissues, they are effectively neutralized in the lung. Chrysotile was found to be rapidly removed from the lung. Fibers longer than 20 microm were cleared with a half-time of 1.3 days, most likely by dissolution and breakage into shorter fibers. Shorter fibers were also rapidly cleared from the lung with fibers 5-20 microm clearing even more rapidly (T1/2 = 2.4 days) than those < 5 microm in length (T1/2 weighted = 23. days). Breaking of the longer fibers would be expected to increase the short fiber pool and therefore could account for this difference in clearance rates. The short fibers were never found clumped together but appeared as separate, fine fibrils, occasionally unwound at one end. Short free fibers appeared in the corners of alveolar septa, and fibers or their fragments were found within alveolar macrophages. The same was true of fibers in lymphatics, as they appeared free or within phagocytic lymphocytes. These results support the evidence presented by McDonald and McDonald (1997) that the chrysotile fibers are rapidly cleared from the lung in marked contrast to amphibole fibers which persist. PMID- 16036746 TI - Chemoprevention of tobacco smoke-induced lung tumors by inhalation of an epigallocatechin gallate (EGCG) aerosol: a pilot study. AB - We investigated whether inhalation of aerosolized epigallocatechin gallate (EGCG) would prevent the development of lung tumors produced by tobacco smoke (TS). Male strain A/J mice were exposed for 5 mo, 6 h/day, 5 days/wk, to a mixture of tobacco sidestream and mainstream smoke. At the end of this exposure, 3 groups were formed: (a) mice exposed to TS and left undisturbed in air; (b) animals exposed to TS and given EGCG aerosol by nose-only inhalation for 30 min per session; and (c) animals exposed to TS and then exposed by nose-only inhalation to water aerosol without any EGCG (sham-exposed group). Three similar groups were formed from animals that previously had been kept in filtered air. In experiment 1, the EGCG concentration in the aerosol was 80 microg/L and administered 3 times a week and in experiment 2 it was 191 microg/L administered twice a week. Inhalation of EGCG did not modulate TS-induced tumorigenesis. In two accompanying positive control experiments, animals treated with the tobacco-specific carcinogen NNK [4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone] were given the same EGCG or water aerosol treatment. In both experiments, EGCG aerosol significantly reduced lung tumor multiplicity by 20% to 30% However, exposure of NNK-treated animals to water solvent alone (sham exposure) produced an even greater reduction in tumor multiplicities (40%). A reduction of lung tumor multiplicities was also observed in animals exposed nose-only once or five times a week to either water aerosols or to filtered air. It is concluded that water soluble chemopreventive agents that need to be ingested in comparatively high doses are not the most suitable candidates for administration by inhalation. PMID- 16036747 TI - Development of a physiologically based pharmacokinetic model for decane, a constituent of jet propellent-8. AB - Decane, a 10-carbon n-alkane and one of the highest vapor phase constituents of jet propellent-8 (JP-8), was selected to represent the semivolatile fraction for the initial development of a physiologically based pharmacokinetic (PBPK) model for JP-8. Rats were exposed to decane vapors at time-weighted average concentrations of 1200, 781, or 273 ppm in a 32-L Leach chamber for 4 h. Time course samples for 1200 ppm and end-of-exposure samples for 781 and 273 ppm decane exposures were collected from blood, brain, liver, fat, bone marrow, lung, skin, and spleen. The pharmacokinetics of decane could not be described by flow limited assumptions and measured in vitro tissue/air partition coefficients. A refined PBPK model for decane was then developed using flow-limited (liver and lung) and diffusion-limited (brain, bone marrow, fat, skin, and spleen) equations to describe the uptake and clearance of decane in the blood and tissues. Partition coefficient values for blood/air and tissue/blood were estimated by fitting end-of-exposure pharmacokinetic data and assumed to reflect the available decane for rapid exchange with blood. A portion of decane is speculated to be sequestered in "deep" pools in the body, unavailable for rapid exchange with blood. PBPK model predictions were adequate in describing the tissues and blood kinetics. For model validation, the refined PBPK model for decane had mixed successes at predicting tissue and blood concentrations for lower concentrations of decane vapor, suggesting that further improvements in the model may be necessary to extrapolate to lower concentrations. PMID- 16036748 TI - Effects of concentrated ambient particles on airway responsiveness and pulmonary inflammation in pulmonary hypertensive rats. AB - Epidemiological studies have associated particulate air pollution with exacerbation of lung function in human populations. However, the relationship between ambient particles and lung function in animal studies has been inconsistent. In order to investigate the effects of concentrated ambient particles (CAPs) on airway responsiveness, we exposed pulmonary hypertensive rats to CAPs using particle concentrator at an EPA of Taiwan supersite, located at a traffic busy urban area nearing Taipei city. The exposure group (n = 5) was exposed to CAPs for 6 h each day for 3 consecutive days (mean mass concentration = 371.7 microg/m(3)), while a control group (n = 6) was exposed to HEPA-filtered air. Whole-body barometric plethysmography was used to measure respiratory frequency, tidal volume, and airway responsiveness before and after exposure. Enhanced pause (Penh) was used as an indicator of airway responsiveness. To improve the accuracy of airway responsiveness measurement, we controlled temperature and humidity. Further, airway responsiveness was determined 5 h after particle exposure to overcome the stress effect in nose-only exposure chambers. After CAPs exposure, we found decreased respiratory frequency and increased tidal volume (p < .05). Using the methacholine challenge test, a significant difference of Penh measured before and after experiment was observed in the CAPs group (p < .05), but not in the filtered air group. Further analysis showed that the Penh difference before and after exposure in the CAPs group was significantly greater than that in the filtered air group (p < .05). We conclude that CAPs could induce airway hyperresponsiveness in pulmonary hypertensive rats. PMID- 16036749 TI - Patterns of histopathological change determined by the point counting method and its application for the hazard assessment of respirable dust. AB - We evaluate the morphometric point counting method (PCM) for qualitatively analyzing pulmonary inflammation and collagen deposits (i.e., fibrosis) in the assessment of the biological hazards of inhaled respirable particles at a realistic dose comparable to that of exposure in the work environment. Rats were exposed by intratracheal instillation to a 2-mg dose, which is close to the estimated overdose at which macrophage clearance is impared, of each of 3 kinds of particulate matter: crystalline silica, crocidolite asbestos, and titanium dioxide. The lung tissue was evaluated at 3 days, 1 wk, and 1, 3, and 6 mo after exposure. Digital images taken of the lung tissue after processing and staining of the lung sections were examined by the PCM under light microscopy. Evidence of inflammation along with progressive inflammatory changes occurred with crystalline silica and crocidolite, which are well-known hazardous particle types. In contrast, lung tissue from rats exposed to titanium dioxide particles demonstrated a decreasing pattern of histopathological change with increasing retention time. Differences in repair patterns of TiO(2) versus crocidolite and silica following the 2-mg dose exposure suggest that the PCM scoring system may be a useful and sensitive tool for qualitatively evaluating the biological hazards of new particle types, for which no toxicological information exists for low-dose exposure, by using the results from assessment of fibrogenic particle types (such as crocidolite and crystalline silica) as well as particle types with low toxicity (such as TiO(2)) as reference points. PMID- 16036750 TI - Extrapolation of the carcinogenic potency of fibers from rats to humans. AB - In 1999 Berry published a model for mesothelioma incidence following fiber exposure. He concluded, that the influence of the solubility of fibers on the mesothelioma rate is 17 times higher in humans than in rats. This conclusion may be helpful for evaluating the carcinogenic risk from man-made vitreous fibers, but it had little influence on some recent discussions. It has been demonstrated using this model, that in an injection experiment with rats, fibers with elimination constants of 0.1/year and 1/year--which would approximately correspond to crocidolite and perhaps ceramic fibers--differ in their mesothelioma risk only by a ratio of 3.2:1. In contrast, for humans exposed continuously from age 20 to age 60 a risk ratio of 4,750:1 is obtained. This result may be helpful for the assessment of the human cancer risk e.g., from exposure to refractory ceramic fibers. However, uncertainty is large, since the life-span of rats is too low to measure the elimination rate of bio-persistent fibers sufficiently. PMID- 16036751 TI - DNA methylation, cell proliferation, and histopathology in rats following repeated inhalation exposure to dimethyl sulfate. AB - Dimethyl sulfate (DMS) is an alkylating agent that is carcinogenic to the respiratory tract of rodents. DNA adducts, cell proliferation, and histopathology were assessed in rats to better understand the molecular dosimetry and tissue dynamics associated with repeated inhalation exposure to DMS. For DNA methylation, rats were exposed to DMS vapor 6 h/day for up to 10 days to 0.0, 0.1, 0.7 and 1.5 ppm. N7-Methylguanine and N3-methyladenine were detected in neutral thermal hydrolysates of DNA isolated from respiratory tract tissues by high-performance liquid chromatography (HPLC) using fluorescence and ultraviolet (UV) detection. DNA methylation was greatest in DNA isolated from nasal respiratory mucosa, less in olfactory, and little was found in lung. N7 Methylguanine levels in respiratory mucosa approached steady-state levels by day 5, and N7-methylguanine persistence following exposure for 5 consecutive days was also determined. Loss of N7-methylguanine from respiratory and olfactory mucosa appeared to follow first-order kinetics. N3-Methyladenine levels were at or below detection limits in all samples. The effect of DMS on histopathology and cell proliferation in the nasal epithelium was also investigated. Rats were exposed nose-only for 2 wk to DMS vapor at concentrations of 0, 0.1, 0.7, or 1.5 ppm. Inhalation exposure to DMS induced degenerative and inflammatory changes in nasal epithelium at >or=0.7 ppm. Cell proliferation evaluations showed a trend towards an increased response at 1.5 ppm. These experiments demonstrate that DMS can induce cytotoxic and proliferative effects and is a potent methylating agent of the nasal mucosa in vivo. These experiments will provide data for the development of dosimetry models useful for risk extrapolation. PMID- 16036752 TI - Interspecies dose extrapolation for inhaled dimethyl sulfate: a PBPK model-based analysis using nasal cavity N7-methylguanine adducts. AB - Dimethyl sulfate (DMS) is a volatile sulfuric acid ester used principally as a methylating agent in a wide variety of industrial applications. DMS reacts with organic macromolecules by a SN2 mechanism. The weight of experimental evidence suggests that DMS possesses genotoxic and carcinogenic potential. Inhalation studies have shown that repeated exposure to DMS leads to tumors in the nasal cavity and lower respiratory tract in both rats and mice. Here we present a quantitative assessment for cross-species dose extrapolation for inhaled DMS using a physiologically based pharmacokinetic (PBPK) model. The model is designed to simulate N7-methylguanine (N7 mG) DNA adduct levels in the nasal mucosa following DMS exposure in rats and humans. This model was parameterized and predictions were tested by comparison against experimentally measured N7 mG DNA adduct levels in rat nasal mucosa following inhalation exposure to DMS. The model based interspecies dose comparison, using N7 mG adduct levels in the nasal respiratory tissue as the appropriate dose metrics, predicts a dose rate seven times higher in rats compared to humans. PMID- 16036753 TI - Expression of C-reactive protein in human lung epithelial cells and upregulation by cytokines and carbon particles. AB - C-reactive protein (CRP) is the prototypic human acute-phase protein and is found at increased levels in the blood during episodes of inflammation. CRP was generally thought to be produced only by hepatocytes; however, several studies have shown extrahepatic synthesis of CRP. A previous study showed that PM10 and ultrafine carbon black (ufCB) were able to induce CRP expression in A549 cells. This study aims to examine the factors that lead to the production of CRP in A549 cells. A549 human lung epithelial cells were treated with cytokines (interleukin 6, tumor necrosis factor alpha, interferon gamma, or interleukin 1beta) or carbon particles (CB and ufCB) for 18 h. It was found that CRP could be expressed within the cells and that CRP was secreted from the cells particularly with tumor necrosis factor alpha, CB and ufCB treatments. It was also found that this expression of CRP with CB and ufCB treatments was dependent on nuclear factor kappa B (NFkappaB). The expression of CRP in A549 cells may indicate an important role for CRP expression and secretion from lung epithelial cells in response to inflammatory stimuli. PMID- 16036754 TI - Assessment of the elution of charcoal, cellulose acetate, and other particles from cigarettes with charcoal and activated charcoal/resin filters. AB - This experiment was designed to study the release of cellulose acetate fibers, charcoal, and other particles from cigarettes with charcoal and activated charcoal/resin filters. For the first time in such studies, efforts were made to identify the particles that were eluted using other analytical techniques in addition to light microscopy. Other corrective measures were also implemented. During the studies it was found that trimming of larger filters to fit smaller filter housings introduced cellulose acetate-like particles from the fibers of the filter material. Special, custom made-to-fit filters were used instead. Tools such as forceps that were used to retrieve filters from their housings were also found to introduce fragments onto the filters. It is believed that introduction of such debris may have accounted for the very large number of cellulose acetate and charcoal particles that had been reported in the literature. Use of computerized particle-counting microscopes appeared to result in excessive number of particles. This could be because the filter or smoke pads used for such work do not have the flat and level surfaces ideal for computerized particle-counting microscopes. At the high magnifications that the pads were viewed for particles, constant focusing of the microscope would be essential. It was also found that determination of total particles by using extrapolation of particle count by grid population usually gave extremely high particle counts compared to the actual number of particles present. This could be because particle distributions during smoking are not uniform. Lastly, a less complex estimation of the thickness of the particles was adopted. This and the use of a simple mathematical conversion coupled with the Cox equation were utilized to assess the aerodynamic diameters of the particles. Our findings showed that compared to numbers quoted in the literature, only a small amount of charcoal, cellulose acetate shards, and other particles are released. It was also shown that those particles would have a low likelihood of reaching the lung. PMID- 16036755 TI - Quantitative analysis of asbestos burden in a series of individuals with lung cancer and a history of exposure to asbestos. AB - Asbestos is recognized as a lung carcinogen. In the present study, tissue from 20 individuals who died from lung cancer and who had a history of exposure to asbestos was evaluated for the presence of asbestos bodies and uncoated asbestos fibers. A digestion procedure was used to isolate the particulates from the tissue. The samples were evaluated by light microcopy to quantify the numbers of ferruginous bodies in the tissue. The uncoated fibers (which included all fibers equal to or greater than 0.5 microm) were analyzed by analytical transmission electron microscopy. Seventeen of the 20 cases were positive for ferruginous bodies (which were morphologically consistent with asbestos bodies). Five of these were found to have concentrations within the range used in our laboratory for the general population (<20 ferruginous bodies/g wet tissue). Nineteen of the 20 cases were found to have asbestos fibers in the higher magnification scan (either 16 K or 20 K). Some of the asbestos fibers identified were specific for the types of exposures that were reported. Most individuals in this study were found to have mixed populations of asbestos fibers in the lung tissue. This suggests that when there are exposures to products containing commercial asbestos there are likely exposures to dust containing noncommercial asbestos. A contrast exists in the dust burden within the lung of these individuals as compared to samples from the general population in that occupational or "occupational-like" exposures such as in these cases are often reflected by the presence of longer fibers of asbestos in the tissue. PMID- 16036756 TI - Pro-oxidative interactions of cobalt with human neutrophils. AB - The primary objectives of this study were to investigate the effects of cobalt(II) chloride (Co, 1.5-25 microM) on the reactivity of hydrogen peroxide (H2O2, 100 microM) or oxidants generated by activated human neutrophils. The prooxidative interactions of Co with H2O2 or cells were measured by luminol enhanced chemiluminescence (LECL), and according to the extent of oxidative inactivation of added alpha-1-proteinase inhibitor (API). Cobalt dramatically potentiated the oxidation of luminol and API by both H2O2 and neutrophils activated with phorbol 12-myristate 13-acetate (5 ng/ml), without affecting the assembly of NADPH oxidase or the magnitude of oxygen consumption by the cells. Using 5,5-dimethyl-pyrolline 1-oxide-based electron spin resonance spectroscopy we were unable to detect hydroxyl radical formation by Co in the presence of either H2O2 or activated neutrophils, while the corresponding LECL responses were unaffected by the hydroxyl radical scavengers benzoate and mannitol (50 mM). These observations indicate that Co potentiates the reactivity of neutrophil derived oxidants, primarily H2O2, which if operative in vivo during exposure to the heavy metal may pose the risk of oxidant- and protease-mediated tissue injury. PMID- 16036757 TI - Performance of a portable whole-body mouse exposure system. AB - A mobile whole-body exposure system was developed for exposing mice to concentrated ambient particulate matter smaller than 2.5 microm in mass median aerodynamic diameter (MMAD). Each 20-L exposure cage was designed to hold 9 mice within individual compartments. This allowed for transport and subsequent exposure. Airflow mixing and the potential for stagnant areas within the compartments were modeled using computational fluid dynamic modeling (CFD). CFD analysis showed no stagnant areas and good mixing throughout the exposure cage. The actual performance of the exposure system was determined for 0.5 to 2.0 microm diameter aerosols by measuring (1) uniformity of aerosol distribution and (2) particle deposition in the tracheobronchial and pulmonary regions of mice exposed in the system. A 0.6-microm MMAD (GSD=2.0) cigarette smoke aerosol was used to experimentally measure the uniformity of aerosol distribution to the nine individual compartments. The average data from three runs showed no statistically significant difference among individual compartments. Particle deposition efficiency in adult male BALB/c mice was measured after exposure (30 min) in the system using monodisperse fluorescent polystyrene latex particles (0.5, 1, and 2 microm aerodynamic diameter). The measured deposition efficiency in this mobile exposure system for the combined tracheobronchial and pulmonary regions of the adult male BALBc mice was 21% for 0.5 microm, 11% for 1.0 microm, and 6.5% for 2.0 microm particles. These deposition efficiencies are similar to those reported for mice exposed in a nose-only exposure system, which indicates that particle losses to animal fur and exposure system surfaces were acceptable. PMID- 16036765 TI - Insights into the origin of postmortem ethanol. AB - Accurate interpretation of the blood ethanol (EtOH) concentration at the time of death presents a difficult task since the origin of detected EtOH in postmortem cases (either in corpses or in specimens after sample collection) may vary. Headspace gas chromatography is the choice method for detecting EtOH in blood or other specimens, due to the accuracy and sensitivity it provides. Possible sources of postmortem EtOH have been the ante-mortem ingestion, the ante-mortem endogenous production and the postmortem microbial neo-formation, which has been considered the most critical factor that could complicate the results. It has been reported that EtOH could be formed postmortem in variable and non predictable amounts, as a function of the type and number of microorganisms present either in corpses or specimens collected at autopsy. The presence of other volatiles-mostly n-propanol-has been correlated to microbial EtOH production, although the quantitative pattern between them and EtOH still remains obscure. The factors most frequently implicated in the mechanism of postmortem EtOH production in corpses have been considered the number and nature of microbes present, the availability of various types of substrates, the temperature and the time. Complication in the interpretation of blood alcohol concentration could arise due to the atypical distribution of EtOH in the body compartments after death. Specimens to blood EtOH ratios reported in the literature are presented. All the aforementioned aspects are discussed in a comprehensive way, providing a deep insight into this essential problem. PMID- 16036766 TI - Weak inhibitors protect cholinesterases from stronger inhibitors (dichlorvos): in vitro effect of tiapride. AB - Metoclopramide is a benzamide dopamine receptor antagonist and serotonine receptor agonist widely used as an antiemetic and gastric prokinetic drug. In addition, metoclopramide is a weak and reversible inhibitor of cholinesterases. The authors have previously shown that metoclopramide has a cholinesterase protective effect against inhibition by organophosphates (OPs). The putative mode of protective action of metoclopramide is, when administered in excess, competion for the active site of the enzyme with the more potent OP. In the present paper the authors present their results using another benzamide with weak cholinesterase inhibitory properties, tiapride (TIA). The purpose of the study was to quantify in vitro the extent of TIA-conferred protection, using dichlorvos (dichlorovinyl dimethyl phosphate; DDVP) as an inhibitor. DDVP is a moderately toxic (LD50 in rats in the milligram range), non-neuropathic OP. The substance is responsible for a large number of accidental or suicidal exposures. Red blood cell (RBC) acetylcholinesterase (AChE) activities in whole blood and butyrylcholinesterase (BChE) activities in human plasma were measured photometrically in the presence of different DDVP and TIA concentrations and IC50 was calculated. Determinations were repeated in the presence of increasing TIA concentrations. The IC50 of DDVP increases with the TIA concentration in a linear manner. The protective effect of TIA on cholinesterase could be of practical relevance in the treatment of OP poisoning. The authors conclude that in vivo testing of TIA as an OP protective agent is warranted. PMID- 16036767 TI - Direct dosing of preweaning rodents in toxicity testing and research: deliberations of an ILSI RSI Expert Working Group. AB - Laboratory animal studies designed to assess the effects of exposure of a test substance during postnatal development are commonly utilized in basic research and to evaluate potential hazard to children for chemical and pharmaceutical regulation. Direct dosing, defined here as the administration of a test substance directly to a preweaning mammal, has been identified as a useful tool that can be used in the conduct of such studies for regulatory purposes. The International Life Sciences Institute Risk Science Institute (ILSI RSI) convened an Expert Working Group to develop guidance on the design and implementation of direct dosing regulatory studies on preweaning mammals, which was published as an ILSI monograph in 2003 (Zoetis and Walls, Principles and Practices for Direct Dosing of Pre-Weaning Mammals in Toxicity Testing and Research, Washington, DC: ILSI Press, 2003). A summary of the Working Group conclusions regarding direct dosing studies with laboratory rodents are presented here, although the ILSI monograph also includes rabbits, canines, swine and nonhuman primates. Issues to be considered when designing the protocol include selection of the test species, the route of administration, dose levels, and the timing of dosing. Knowledge of the maturational status of the test species and information on critical windows of development are important in creating a valid study design. Most common routes of administration (e.g., oral, inhalation, injection) are possible with typical laboratory species; however, adjustments may be necessary due to practical considerations. Information on the pharmacokinetic profile in young animals versus adults and in the test species versus humans is very useful for determining dosing parameters. The conduct of the study and the interpretation of the data will be improved by an understanding of confounding factors as well as statistical and biological issues specific for postnatal studies. Ultimately, the success of the study will depend upon careful preparation, including thorough training of the technical staff. PMID- 16036768 TI - A preliminary assessment of the acute and subchronic toxicity profile of phase2: an alpha-amylase inhibitor. AB - Phase2, which has been reported to reduce body weight by its inhibition of a amylase, was evaluated for toxicity in young adult male and female Wistar rats (10 animals/dose group). Evaluations included mortality, change in body weight, food consumption pattern, organ weight, and other adverse side reactions as well as hematological, biochemical, and histopathological analyses. Acute toxicity was determined after a single dose of Phase2 by oral gavage at doses of 5.0, 1.0, and 0.5 g/kg body weight. Animals were sacrificed on fourteen days after Phase2 administration. Subchronic toxicity was determined by administering Phase2 daily for 90 days to rats, at doses of 1.0, 0.5, and 0.2 g/kg body weight. These animals were sacrificed on day 90. Acute and subchronic administration of Phase2 did not produce any adverse reactions or any significant change in the loss of body weight as compared to untreated controls, organ weight, and mortality. Administration of Phase2 did not alter the hepatic and renal function, and did not produce any change in the hematological parameters and in lipid profile. Subchronic administration produced a reduction in the food consumption after 77 days (1.0 g/kg body weight). These data indicate that acute and subchronic administration of Phase2 did not produce any toxicity to rats as evident from weight change, mortality, and limited biochemical and histopathological analyses. PMID- 16036769 TI - Phenylacetaldehyde oxidation by freshly prepared and cryopreserved guinea pig liver slices: the role of aldehyde oxidase. AB - Phenylacetaldehyde is formed when the xenobiotic and biogenic amine 2 phenylethylamine is inactivated by a monoamine oxidase-catalyzed oxidative deamination. Exogenous phenylacetaldehyde is found in certain foodstuffs such as honey, cheese, tomatoes, and wines. 2-Phenylethylamine can trigger migraine attacks in susceptible individuals and can become fairly toxic at high intakes from foods. It may also function as a potentiator that enhances the toxicity of histamine and tyramine. The present investigation examines the metabolism of phenylacetaldehyde to phenylacetic acid in freshly prepared and in cryopreserved guinea pig liver slices. In addition, it compares the relative contribution of aldehyde oxidase, xanthine oxidase, and aldehyde dehydrogenase in the oxidation of phenylacetaldehyde using specific inhibitors for each oxidizing enzyme. The inhibitors used were isovanillin for aldehyde oxidase, allopurinol for xanthine oxidase, and disulfiram for aldehyde dehydrogenase. In freshly prepared liver slices, phenylacetaldehyde was converted mainly to phenylacetic acid, with traces of 2-phenylethanol being present. Disulfiram inhibited phenylacetic acid formation by 80% to 85%, whereas isovanillin inhibited acid formation to a lesser extent (50% to 55%) and allopurinol had little or no effect. In cryopreserved liver slices, phenylacetic acid was also the main metabolite, whereas the 2 phenylethanol production was more pronounced than that in freshly prepared liver slices. Isovanillin inhibited phenylacetic acid formation by 85%, whereas disulfiram inhibited acid formation to a lesser extent (55% to 60%) and allopurinol had no effect. The results in this study have shown that, in freshly prepared and cryopreserved liver slices, phenylacetaldehyde is converted to phenylacetic acid by both aldehyde dehydrogenase and aldehyde oxidase, with no contribution from xanthine oxidase. Therefore, aldehyde dehydrogenase is not the only enzyme responsible in the metabolism of phenylacetaldehyde, but aldehyde oxidase may also be important and thus its role should not be ignored. PMID- 16036770 TI - Effects of postnatal exposure to a mixture of polychlorinated biphenyls, p,p' dichlorodiphenyltrichloroethane, and p-p'-dichlorodiphenyldichloroethene in prepubertal and adult female Sprague-Dawley rats. AB - The postnatal period is a critical phase of development and a time during which humans are exposed to higher levels of persistent organic pollutants (POPs), than during subsequent periods of life. There is a paucity of information describing effects of postnatal exposure to environmentally relevant mixtures of POPs, such as polychlorinated biphenyls (PCBs), p,p'-dichlorodiphenyltrichloroethane (DDT), and p,p'-dichlorodiphenyldichloroethene (DDE). To provide data useful for the risk assessment of postnatal exposure to POPs, mixtures containing 19 PCBs, DDT, and DDE were prepared according to their concentrations previously measured in the milk of Canadian women, and dose-response effects were tested on the proliferation of MCF7-E3 cells in vitro, and in vivo experiments. Female neonates were exposed by gavage at postnatal days (PNDs) 1, 5, 10, 15, and 20 with dosages equivalent to 10, 100, and 1000 times the estimated human exposure level over the first 24 days of life. The MCF7-E3 cells showed a 227% increase in the AlamarBlue proliferation index, suggesting estrogen-like properties of the mixture, but this was not confirmed in vivo, given the absence of uterotrophic effects at PND21. An increase (511%) in hepatic ethoxyresorufin-o-deethylase activity at the dose 100 x was the most sensitive endpoint among those measured at PND21 (organ weight, mammary gland and ovarian morphometry, hepatic enzyme inductions, serum thyroxine and pituitary hormones). In liver samples from older female rats (previously involved in a mammary tumor study [Desaulniers et al., Toxicol. Sci. 75:468-480, 2001]), hepatic metabolism of 14C-estradiol-17beta (E2) at PND55 to PND62 was significantly higher in the 1000x compared to the control group, but hepatic detoxification enzyme activities had already returned to control values. The production of hepatic 2-hydroxy-E2 decreased, whereas that of estrone increased with age. In conclusion, the smallest dose of the mixture to induce significant effects was 100x, and mixture-induced changes in the hepatic metabolism of estrogens might be a sensitive indicator of persistent effects. PMID- 16036771 TI - The mourning of an enlarged and diseased prostate. PMID- 16036772 TI - Hugh Hampton Young-father of modern American urology. PMID- 16036773 TI - Laparoscopic assisted placement of peritoneal dialysis catheters for selected patients with previous abdominal operation. AB - Peritoneal dialysis is an established alternative method for the management of patients with end-stage renal disease. Recently, laparoscopy has been utilized in assisting the insertion of catheters under direct vision. The efficacy of the laparoscopic approach for patients with a history of abdominal surgery remains largely unknown. The purpose of this study is to evaluate laparoscopy in the placement of peritoneal dialysis catheters for selected patients with previous abdominal operation. Laparoscopic assisted placement of peritoneal dialysis catheters was performed in 20 patients, who were carefully selected preoperatively and who also underwent previous abdominal operation between April 1999 and July 2001. Previous abdominal operation included appendectomy, ovarian resection, hysterectomy, cesarean section, open cholecystectomy, segmental resection of the small intestine, and truncal vagotomy with pyloroplasty. The procedure was performed using two 10-mm and one 5-mm abdominal trocar. All of the patients tolerated this procedure without significant surgical complications. However, 3 patients developed temporary hemoperitoneum, and 1 patient developed dialysate leakage. The overall success rate of catheter function (> 30 days after laparoscopy) was 90%, except in 2 cases where the catheter functioned poorly due to severe intra-abdominal adhesions. Simultaneous laparoscopic adhesiolysis was successfully performed in 5 cases. Laparoscopic implantation of peritoneal dialysis catheters appears to be a straightforward procedure, even for patients with previous abdominal operation. We believe that this technique may extend the application of peritoneal dialysis treatment in patients with previous abdominal surgery after discreet evaluation preoperatively. PMID- 16036774 TI - Influence of pneumoperitoneum on small bowel anastomoses: a histological analysis in the rat model. AB - Laparoscopic techniques are increasingly applied for the treatment of diverse gastrointestinal diseases. With regard to reports of a pronounced decrease of intra-abdominal blood flow with increasing intra-abdominal pressure, the present study investigates the impact of pressure and gas type on ischemia in small bowel anastomoses in the rat model. Laparotomy and ileoileal anastomosis were performed in 39 male Sprague-Dawley rats. A CO2 or helium pneumoperitoneum of 3 mm Hg or of 6 mm Hg was maintained before and after anastomoses. Rats in the control group received no pneumoperitoneum. Animals were sacrificed after 5 d, and the anastomotic region was explanted for subsequent histopathological examinations. In hematoxylin and eosin (HE)-stained sections, the Chiu score, villi configuration, and number of goblet cells were analyzed. Proliferation (Ki67) and expression of a matrix metalloproteinase (MMP-8) were examined by immunohistochemistry. Mucosal damage according to the scoring system by Chiu, the number of goblet cells, the villus length, the proliferation (Ki67), and the submucosal expression of MMP-8 was similar in all groups. Our results suggest that within a certain range of pressures and time, laparoscopic assisted surgery using CO2 pneumoperitoneum can be performed safely. Helium gas offers no advantages over CO2. PMID- 16036775 TI - Differential effect of tacrolimus on dermal and intestinal wound healing. AB - Tacrolimus, used in organ transplantation, inhibits cellular immune function. Little is known about the effect on dermal and colonic healing. Groups of 10 rats underwent dorsal skin incision, and polyvinyl alcohol sponges were implanted subcutaneously. Beginning at the day of wounding, rats were treated intraperitoneal with 1.0 or 2.0 mg tacrolimus/kg/day. Animals were sacrificed 10 d later to determine wound breaking strength and reparative collagen deposition. Expression of transforming growth factor (TGF)-beta, tumor necrosis factor (TNF) alpha, and interferon (IFN)-gamma was studied in wounds. Groups of 8 rats underwent laparotomy and left colonic anastomosis. These rats were treated by subcutaneous injections with 2.0 or 5.0 mg tacrolimus/kg. Animals were sacrificed 5 d later to test colonic bursting pressure and reparative collagen deposition. Expression of TGF-beta, TNF-alpha, IFN-gamma, and CD4 and CD8 in the anastomosis was investigated. Tacrolimus impaired dermal healing (p < .05). This was paralleled by decreased expression of TGF-beta (stimulates healing) and increased expression of IFN-gamma and TNF-alpha (both inhibit healing) (p < .05). In contrast, tacrolimus did not inhibit healing of colonic anastomoses. No effect was seen on the expression of TGF-beta, TNF-alpha, IFN-gamma, and CD4 and CD8 in colonic anastomoses. We concluded that tacrolimus differentially effects tissue healing and expression of cellular mediators in dermal and intestinal wounds. PMID- 16036776 TI - Thoracotomy is associated with significantly more profound suppression in lymphocytes and natural killer cells than video-assisted thoracic surgery following major lung resections for cancer. AB - Major surgery is immunosuppressive, and this could have an impact on postoperative tumor immunosurveillance and, therefore, long-term survival in cancer patients. Video-assisted thoracic surgery (VATS) lung resection is a new alternative surgical approach to thoracotomy for patients with early lung cancer. This is a pilot study to examine the postoperative changes in leukocytes, lymphocyte subsets, B cells, T cells, and natural killer (NK) cells in non-small cell lung cancer (NSCLC) patients undergoing lung resection with VATS versus thoracotomy approaches. Twenty-one consecutive patients with resectable primary NSCLC were assigned to VATS or thoracotomy approach over a 3-month period. Blood samples were collected preoperatively and at postoperative days (POD) 1, 3, and 7 for flow cytometry determination of total leucocytes, B cells, NK cells, lymphocytes, total T cells, and T4 and T8 cell numbers. There were no demographic differences between the two groups. Compared with the preoperative values, significantly increased total white cell numbers were detected at POD 1, 3, and 7 in all patients. At POD 1, although T8 cells and NK cells were reduced in both groups, total T cell, T4 cell, and lymphocyte numbers were significantly reduced only in the thoracotomy group. At POD 7, NK cell numbers were significantly lower in the thoracotomy group than that in the VATS group. No significant intra- or intergroup differences were seen with B cells. No significant differences in survival or disease-free survival were found between the two groups. Thus, VATS major lung resection for NSCLC is associated with less, as well as quicker recovery from, postoperative immunosuppression compared with the thoracotomy approach. The clinical relevance of better preserved cellular immunity in the early postoperative period warrants confirmation from large randomized trials. PMID- 16036777 TI - Comparison of adhesive properties of five different prosthetic materials used in hernioplasty. AB - This experimental study was designed to assess and to compare intra-abdominal adhesions following the use of five commercially available prosthetic mesh grafts in the repair if abdominal wall defects. Sixty Wistar albino rats were randomly divided into six groups (n = 10). A 2 x 1 cm defect at abdominal wall was created and defects were closed either primarily or with one of the following prosthetic mesh grafts: monofilament polypropylene, polytetrafluoroethylene, sodium hyaluronate/carboxymethylcellulose-coated polypropylene, polypropylene/polyglactin 910 composite, or resorbable hydrophilic collagen coated multifiber polyester. The severity of adhesions was graded, tensile strengths of adhesions were measured, and histopathological grades of inflammation and fibrosis were evaluated. Polypropylene mesh resulted in more adhesion formation in comparison to primary repair and other grafts used in this study, except polypropylene/polyglactin 910 composite mesh. In addition, the highest tensile strength of omental adhesions was detected in the polypropylene group (chi2 = 26.249; p = .0001). Polyester composite mesh caused the least adhesion formation among the groups. Sodium hyaluronate/carboxymethylcellulose coated polypropylene and polyester composite meshes revealed the highest fibrosis scores (chi2 = 50.776; p = .0001). The highest inflammatory activity was detected in the polytetrafluoroethylene mesh group (chi2 = 16.564; p = .005). Thus, sodium hyaluronate/carboxymethylcellulose-coated polypropylene and polytetrafluoroethylene meshes following polyester composite mesh were the minimal adhesion-forming grafts in this study. Disadvantages of the polytetrafluoroethylene mesh were lower fibrotic activity and higher inflammatory reaction to the graft. PMID- 16036778 TI - The role of fibrin tissue adhesives in flap necrosis in rats. AB - Flap necrosis is an important issue in surgery, and fibrin tissue adhesives, due to beneficial properties in preventing flap necrosis, were used in this study. Two groups, each comprising of 10 rats, were formed. Group I served as a control group, and fibrin tissue adhesive was applied to group II. The fibrinogen and thrombin concentrations in fibrin tissue adhesive were 30 mg/ml and 10 U/ml, respectively. The mean area of flap necrosis was 687.5 +/- 72.5 mm2 and 78.5 +/- 11.0 mm2 in the control and fibrin tissue adhesive groups (p < .0001), respectively. The percentage of flap necrosis was significantly lower in the fibrin tissue adhesive group compared to the control group (5.6% vs 49.1%) (p < .0001). Fibrin tissue adhesives decreased flap necrosis significantly compared to the control group. PMID- 16036779 TI - Slow down and still be good in surgery, otherwise ask William Halsted. PMID- 16036780 TI - Daniel Hale Williams: pioneer Black surgeon and educator. PMID- 16036781 TI - Evaluation of Pringle maneuver during liver resection in a rat model of surgical obstructive jaundice. AB - Temporary portal triad clamping (Pringle maneuver) during liver resection reduces intraoperative blood loss. A normal liver can safely tolerate normothermic ischemia for up to 60 min. However, its safety in patients with surgical obstructive jaundice (SOJ) is not known. Therefore, we investigated the effect of hepatic ischemia in an experimental rat model of SOJ created by ligating the bile duct. Four groups of rats were created: Group I (sham operation, 10 days later, liver resection); Group II (sham operation, 10 days later, liver resection with 5 min of hepatic ischemia); Group III (bile duct ligation, 10 days later, liver resection); and Group IV (bile duct ligation, 10 days later, liver resection with 5 min of hepatic ischemia). The ischemic injury was assessed by the survival of rats, liver tissue malondialdehyde and total glutathione (markers of free radical injury), serum alanine aminotransferase, aspartate aminotransferase, and liver histology. The results showed decreased survival (47.6% vs. 90% [p = .046]), increased liver tissue malondialdehyde (161 +/- 35 vs. 129 +/- 33 microg/gm liver tissue [p = .05]), and decreased liver tissue total glutathione (565 +/- 169 vs. 1075 +/- 276 nmol/gm liver tissue [p = .05]) in rats with SOJ subjected to hepatic ischemia when compared to nonjaundiced rats. The changes in serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase showed an increasing trend in the SOJ group but were not statistically significant. Ischemic changes in liver histology were seen more often in the SOJ group but were not statistically significant. These data suggest that temporary portal triad clamping in an experimental model of SOJ is detrimental to the outcome of liver resection. PMID- 16036782 TI - Pedunculated synovium grafts in articular cartilage defects in rabbits. AB - A rabbit model was used to assess the nature of healing tissues in hyaline cartilage defects and to compare the healing in defects treated with pedunculated synovium grafts to those in defects without synovial grafting. Both knees of 28 1 year-old rabbits were operated. A 3 x 2-mm cartilage defect that exposed cancellous bone was created in the non-weight-bearing area of each medial femoral condyle. Each right-knee defect was covered with a pedunculated synovial graft obtained from the same joint, and the left-knee defects were left uncovered as controls. Groups of rabbits were sacrificed at 3, 6, 12, and 24 weeks postsurgery. Sections from each knee were stained with hematoxylin-eosin and safranin O-fast green staining, and were immunohistochemically stained for type II collagen. The healing at each site was histologically scored, and the intensity of staining for type II collagen was graded. At 12 and 24 weeks, statistical comparisons of histological scores revealed significantly more hyaline cartilage tissue in the synovium-grafted defects. At 24 weeks, these same defects showed significantly more type II collagen. Thus, pedunculated synovium transplantation appears to hold promise as a method for repairing hyaline cartilage defects. PMID- 16036783 TI - Role of TIMP-2 in fascia transversalis on development of inguinal hernias. AB - The exact reason for the development of inguinal hernia has not been completely determined. However, it is known that the fascia transversalis (FT) is one of the structures preventing development of hernias. In the etiology of the inguinal hernia, disorders in collagen metabolism have been proposed, and the role of metalloproteinases in remodeling the collagen has recently been of great importance. We could not encounter any study where the role of metalloproteinase inhibitors was evaluated in inguinal hernia. We obtained samples of FT from patients with direct and indirect hernia and used an immunohistochemical method to determine tissue inhibitor of metalloproteinase-2 (TIMP-2) expression. In the study group, samples of FT were taken during the operation from 45 patients, of which 35 were indirect and 10 were direct inguinal hernias. In the control group, samples of FT from various abdominal incisions were also taken from 45 patients with no hernia and operated upon for another pathology. TIMP-2 scores of a direct inguinal hernia were significantly less than those of the control group. However, no difference has been found between the TIMP-2 scores of an indirect inguinal hernia and those of the control group. Decreased TIMP-2 scores in patients with a direct inguinal hernia, compared with both the indirect inguinal hernia group and the control group, explain the reason for the increase in matrix metalloproteinase-2 (MMP-2) that has been proposed in some studies. Therefore, it can be expressed that a decreased activity of TIMP-2 plays a role in inguinal hernia development. PMID- 16036784 TI - The safety and accuracy of sentinel-node biopsy in early-stage invasive breast cancer--Turkish experience. AB - The purpose of this prospective study was to investigate the correlation of sentinel lymph node (SLN) and axillary lymph node (ALN) metastasis in early-stage invasive breast cancer in a single institution. One hundred and fifteen patients with early-stage invasive breast cancer first underwent SLND followed by an appropriate surgical procedure (modified radical mastectomy, lumpectomy + axillary dissection, simple mastectomy + mammoplasty). In this series, a radioactive agent (technetium) was used to investigate the sentinel lymph node/nodes. In 28 (24.3%) patients, metastases were found in both SLN and axillary dissections. There were no metastases in either of these procedures in 69 (60%) patients. SLN metastasis was found in 13 (11.3%) patients, but no axillary metastasis was found. No skip metastasis was detected. Five patients in whom the sentinel node was not found were also negative for axillary metastasis. As the studies progress in this direction, it might be possible to avoid axillary dissection in patients with early breast cancer in whom metastasis in SLN cannot be detected. We believe this will reduce morbidity from breast cancer surgeries. PMID- 16036785 TI - Physioanatomic study of the diaphragmatic crura: the identification of autonomous "gastroesophageal sphincter". AB - It is claimed that the right diaphragmatic crus forms a loop sphincter around the lower esophagus. We investigated the hypothesis that the loop-shaped muscle does not belong to the diaphragmatic crura either anatomically or physiologically and is considered an autonomous muscle. Thirty-two cadaveric specimens (20 males, 12 females, 22 adults, mean age 36.2 +/- 11.8 years, 10 mature neonatal deaths) fixed in 10% formalin were studied anatomically. Electrophysiologic study was performed in 14 subjects (8 men, 6 women, mean age 36.6 +/- 8.2 years) scheduled for laparatomy. The loop muscle was stimulated by needle electrode, and response from the muscle and right crus was recorded by two electrodes. The test was repeated using an electrode into the left crus. Response of the loop muscle to individual stimulation of right and left crus was also registered. Muscle bundles formed a U-shaped loop around the lower esophagus, with the two limbs inserted into the 1st lumbar vertebra. An "esophago-sphincteric space" existed between the two limbs anterior to the esophagus. Three patterns of loop insertion were identified: classic, limb fusion, and limb crossing. The two crura were tendinous from their vertebral attachment, became fleshy, and fanned out proximally to merge with the fleshy diaphragm proper. Stimulation of loop muscle affected significant increase of its electric activity but had no effect on right or left crus. Crural stimulation produced significant increase of their electric activity and no effect on the loop muscle. Muscle fibers surrounding the lower esophagus formed a U-shaped loop. The loop arrangement seems to play significant role in competent mechanism of the gastroesophageal junction. Anatomic and electrophysiologic evidence suggest that the loop muscle is an autonomous muscle that surrounds the lower esophagus and is not derived from the diaphragmatic crura. We call this muscle "striated gastroesophageal sphincter". PMID- 16036786 TI - Effect of a BIOcocktail on the immune response at the early postoperative period in mice. AB - Since human subjects and laboratory animals may develop impaired immune response during surgery and the postoperative period, efforts have been made to preserve normal immune functions following surgery by the administration of nutritional supplements and probiotics. The present study was designed to examine the effect of a new nutritional supplement, BIOcocktail, on immune parameters in mice exposed to surgery. Forty mice were assigned to 4 groups containing 10 animals each. Two control groups (with and without subsequent sham laparotomy) were given tap water for 45 min every day for 2 weeks. The remaining 2 groups, with and without laparotomy, received BIOcocktail given orally for the same period of time. The proliferative response of splenic cells (splenocytes) stimulated with phytohemagglutinin (PHA), concanavalin A (Con A) and lipopolysaccharide (LPS) was determined by [3H]thymidine uptake. Cytokine levels were measured in splenocyte supernatants and sera using enzyme-linked immunosorbent assay (ELISA) kits. Natural killer cell activity of splenocytes was evaluated by 51Cr-release assay. Laparotomy, without BIOcocktail administration, was followed by a decreased proliferative response of splenocytes to PHA, Con A, and LPS and an increase in interleukin (IL)-6 serum level. In addition, a decreased secretion of IL-1beta, IL-12 and tumor necrosis factor (TNF)-alpha by the splenocytes was observed. Mice treated with BIOcocktail before laparotomy maintained a preoperative level of splenocyte proliferative response and serum concentrations of IL-12. It is concluded that BIOcocktail administered to mice for 2 weeks before operation resulted in the preservation of T- and B-cell proliferative response to mitogens and in the prevention of postoperative decrease in IL-12 serum level. PMID- 16036787 TI - You do not have to work hard to be successful and entirely happy. PMID- 16036791 TI - Olfactory transmission of neurotropic viruses. AB - Olfactory receptor neurons are unique in their anatomical structure and function. Each neuron is directly exposed to the external environment at the site of its dendritic nerve terminals where it is exposed to macromolecules. These molecules can be incorporated into by olfactory receptor neurons and transported transsynaptically to the central nervous system. Certain neurotropic pathogens such as herpes simplex virus and Borna disease virus make use of this physiological mechanism to invade the brain. Here the authors review the olfactory transmission of infectious agents and the resulting hazards to human and animal health. PMID- 16036790 TI - Human immunodeficiency virus type 1 genetic diversity in the nervous system: evolutionary epiphenomenon or disease determinant? AB - Over the past decade there has been a revolution in the understanding and care of human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) associated disease. Much of this progress stems from a broader recognition of the importance of differences in viral types, including receptor preference(s), replication properties, and reservoirs, as contributing factors to immunosuppresion and disease progression. In contrast, there is limited conceptualizatin of viral diversity and turnover in the brain and circulation in relation to neurocognitive impairments. Herein, the authors review current concepts regarding viral molecular diversity and phenotypes together with features of HIV-1 neuroinvasion, neurotropism, neurovirulence and neurosusceptiblity. Viral genetic and antigenic diversity is reduced within the brain compared to blood or other systemic organs within individuals. Conversely, viral molecular heterogeneity is greater in patients with HIV-associated dementia compared to nondemented patients, depending on the viral gene examined. Individual viral proteins exert multiple neuropathogenic effects, although the neurological consequences of different viral polymorphisms remain uncertain. Nonetheless, host genetic polymorphisms clearly influence neurological disease outcomes and likely dictate both acquired and innate immune responses, which in turn shape viral evolution within the host. Emerging issues include widespread antiretroviral therapy resistance and increasing awareness of viral superinfections together with viral recombination, all of which are likely to impact on both HIV genetic variation and neuropathogenesis. With the persisting prevalence of HIV-induced neurocognitive disabilities, despite marked improvements in managing immunosuppression, it remains imperative to fully define and understand the mechanisms by which viral dynamics and diversity contribute to neurological disease, permitting the development of new therapeutic strategies. PMID- 16036792 TI - Depressive symptoms, quality of life, and neuropsychological performance in HIV/AIDS: the impact of gender and injection drug use. AB - Limited attention has been paid to the potential impact of gender and injection drug use (IDU) on mood, quality of life, and neuropsychological performance in human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS). Several studies that describe the natural history of HIV/AIDS in terms of mental health and neuropsychological ability have focused solely on men or have excluded injection drug users. Women and injection drug users are two groups for whom the incidence of HIV infection is increasing. Additionally, the National Academy of Sciences recently recommended that studies concerned with health-related research include males and females, and that researchers analyze their data for gender differences. The goals of the current study were to investigate possible relationships between HIV and IDU status and depressive symptoms, quality of life, and neuropsychological performance in women and men matched for age, race, and education. Overall, women reported more depressive symptoms than men, and this gender difference was most evident in women who were both infected with HIV and who were also injection drug users. Women and HIV-infected individuals reported the poorest quality of life scores. Women outperformed men on a measure of verbal memory and HIV(-) participants outperformed HIV(+) participants on a measure of perceptual speed. Finally, gender and HIV status interacted such that uninfected women performed the best, and infected men performed the worst, on a test of verbal memory. A better understanding of how men and women with different drug use profiles respond to HIV/AIDS may substantially improve survival, as well as aspects of daily functioning, of affected individuals. Thus, further study and development of treatment protocols targeted at including women and IDU are needed. PMID- 16036793 TI - Cerebrospinal fluid from human immunodeficiency virus--infected individuals facilitates neurotoxicity by suppressing intracellular calcium recovery. AB - Neurologic decline associated with penetration of human immunodeficiency virus type 1(HIV-1) into the central nervous system is thought to be due, in large part, to inflammation and local secretion of neurotoxic substances. To examine the cellular processes that mediate neurotoxicity in vivo, the authors valuated the ability of neurons to maintain intracellular calcium homeostasis in the presence of toxic cerebrospinal fluid (CSF) (CSF(tox)) collected from a subset of HIV-infected individuals. Exposure of rat neural cultures to CSF(tox) resulted in a gradual increase in intracellular calcium in neurons (+63%), microglia (+251%), and astrocytes (+52%). Pretreatment of neural cultures with CSF(tox) resulted in an exaggerated calcium response to a brief pulse of glutamate and a > 90% suppression of the rate of recovery of intracellular calcium. Attempts to model the deficit using inhibitors of calcium transport across endoplasmic reticulum, mitochondrial, or plasma membrane indicated that blockade of the plasma membrane sodium/calcium exchanger was best able to reproduce the deficits seen during exposure to CSF(tox). Because the inability of cells to maintain calcium homeostasis would lead to exaggerated responses from a wide variety of stimuli, therapeutics designed to facilitate calcium transport from the cell may provide more comprehensive and effective intervention than strategies targeted to specific receptor pathways. PMID- 16036794 TI - Retinoid-induced mu opioid receptor expression by phytohemagglutinin-stimulated U937 cells. AB - Opioid use may be associated with an increased risk of neurological disease in human immunodeficiency virus (HIV) infection through effects on immune cell function. Studies were performed to examine the effects of specific retinoid receptor activation on mu opioid receptor (MOR) production by phytohemagglutinin (PHA)-stimulated U937 cells, a mononuclear cell line. PHA stimulation increased activation of the MOR promoter as well as levels of MOR mRNA, total receptor protein in cell lysates, and surface and cytoplasmic receptor expression. Retinoid X receptor (RXR) agonist and retinoic acid receptor (RAR) antagonist further increased MOR expression by the PHA-stimulated cells. In contrast, MOR expression was suppressed by RAR agonist and by RXR antagonist. Finally, opioid receptor binding was also increased by RXR agonist and RXR antagonist; no increase in binding occurred in the presence of RAR agonists and RXR antagonist. All together, these studies suggest that MOR expression in U937 cells can be differentially regulated by specific retinoid receptor activation. Such effects may have important clinical relevance for opioid users with HIV infection, including individuals with neurological disease. PMID- 16036795 TI - Up-regulation of astrocyte cyclooxygenase-2, CCAAT/enhancer-binding protein homology protein, glucose-related protein 78, eukaryotic initiation factor 2 alpha, and c-Jun N-terminal kinase by a neurovirulent murine retrovirus. AB - In susceptible strains of mice, infection with the mutant retrovirus MoMuLV-ts1 causes a neurodegeneration and immunodeficiency syndrome that resembles human human immunodeficiency virus-acquired immunodeficiency syndrome (HIV-AIDS). In this study the authors show increased expression of cyclooxygenase-2 (COX-2) in the brainstem tissues of ts1-infected mice. Up-regulated central nervous system (CNS) levels of this enzyme are associated with HIV-associated dementia and other inflammatory and neurodegenerative diseases such as amyotrophic lateral sclerosis, Alzheimer's disease, and Parkinson's disease. In brainstem sections, the authors find that astrocytes surrounding spongiform lesions contain increased amounts of immunoreactive COX-2. COX-2 is also up-regulated in cultured ts1 infected cells from the C1 astrocytic cell line, and activation of c-Jun N terminal kinase, or JNK, pathway. Markers of endoplasmic reticulum (ER) stress, specifically the CCAAT/enhancer-binding protein (CHOP), the glucose-related protein 78 (GRP78), and phosphorylated eukaryotic initiation factor 2 alpha (eIF2 alpha), were also up-regulated in ts1-infected C1 astrocytes. Up-regulation of COX-2 and the above ER signaling factors was reversed by treatment of the infected cells with curcumin which specifically inhibits the JNK/c-Jun pathway. These findings indicate that the JNK/c-Jun pathway is most likely responsible for COX-2 expression induced by ts1 in astrocytes, and that ts1 infection in astrocytes may lead to up-regulation of both inflammatory and ER stress pathways in the central nervous system. Because COX-2 inhibitors are now widely used to treat inflammatory conditions in animals and humans, this finding suggests that these drugs may be useful for therapeutic intervention in neurodegenerative syndromes as well. PMID- 16036796 TI - The central nervous system is a viral reservoir in simian immunodeficiency virus- infected macaques on combined antiretroviral therapy: a model for human immunodeficiency virus patients on highly active antiretroviral therapy. AB - This study used a simian immunodeficiency virus (SIV)-macaque model to determine whether virus persists in the central nervous system (CNS) of human immunodeficiency virus (HIV)-infected individuals in which plasma viral load has been suppressed by highly active antiretroviral therapy. SIV-infected macaques were treated with two reverse transcriptase inhibitors: PMPA (q- R-(2 phosphonomethoxypropyl)adenine)which does not cross the blood-brain barrier, and FTC (beta-2('),3(')-dideoxy-3 thia-5-fluorocytidine), which does. Viral DNA and RNA were quantitated in the brain after 6 months of suppression of virus replication in blood and cerebrospinal fluid (CSF). Viral DNA was detected in brain from all macaques, including those in which peripheral viral replication had been suppressed either by antiretroviral therapy or host immune responses. Significant neurological lesions were observed only in one untreated macaque that had active virus replication in the CNS. Expression of the inflammatory markers, major histocmopatibility complex (MHC) II and CD68 was significantly lower in macaques treated with PMPA/FTC. Thus, although antiretroviral treatment may suppress virus replication in the periphery and the brain and reduce CNS inflammation, viral DNA persists in the brain despite treatment. This suggests that the brain may serve as a long-term viral reservoir in HIV-infected individuals treated with antiretroviral drugs that suppress virus replication. PMID- 16036797 TI - High susceptibility of a human oligodendroglial cell line to herpes simplex type 1 infection. AB - More than 20 infectious agents, ranging from retroviruses to mycobacteria, have been associated with multiple sclerosis onset or relapses in which oligodendrocytes, the myelin-forming cells of the central nervous system, are the initial target of the pathogenic status. In this work, the nature of the susceptibility of the human precursor oligodendroglial KG-1C cell line to herpes simplex virus type 1 (HSV-1) was investigated. Infection of KG-1C cells was characterized by a high level of virus production and a notable progression of the cytopathic effect. After infection, there was a significant shut-off of host mRNA translation, which was correlated with evident synthesis of viral proteins. An examination by electron microscopy of the infected cells revealed the presence of large clusters of mitochondria located in the proximity of intracellular HSV-1 particle groups. In addition, transmission electron microscopy and nuclear fluorescence analysis showed neither signs of chromatin condensation nor of apoptotic bodies. Furthermore, procaspase-3 remained uncleaved, suggesting that apoptosis does not take place, at least in this system. Finally, expression and localization of MAL2, a subpopulation of detergent-insoluble lipid raft protein, was studied. Detection of MAL2 significantly increased after infection and it was colocalized with HSV-1 proteins. From these findings the authors conclude that human oligodendrocyte-like cells are highly susceptible to HSV-1 infection. The implications of this for central nervous system viral infection are discussed. PMID- 16036798 TI - Clinicopathological and virological analyses of familial human T-lymphotropic virus type I--associated polyneuropathy. AB - Human T-lymphotropic virus type I (HTLV-I) is known to be the causative agent of the chronic myelopathy, HTLV-I--associated myelopathy (HAM), and on rare occasions infection is also associated with the development of polyneuropathy. Here the authors present an HTLV-I--positive family of whom four members developed a chronic demyelinating polyneuropathy without HAM. Four female patients in a family from Hokkaido in Japan developed distal dominant paresthesia and muscle weakness in the second and third decades of their life. Neurological findings at ages ranging from 50 to 65 years included mild painful sensorimotor disturbances with atrophy of the distal parts of the extremities but without pyramidal signs or hyperactive tendon reflexes. Magnetic resonance imaging (MRI) findings of brain and spinal cord were unremarkable. Serum HTLV-I antibody levels were elevated at 1:8192 to 1:32,768, whereas those in cerebrospinal fluid were low at 1:4 to 1:8. Electrophysiological studies revealed polyphasic compound muscle action potentials with denervation potentials on nerve conduction studies and neurogenic patterns by electromyography, which were consistent with signs of chronic motor dominant demyelinating polyneuropathy. Sural nerve biopsy showed decreased myelinated fibers, occurrence of globule formation, myelin ovoid and remyelinated fibers, and an infiltration of CD68-positive macrophages with occasional CD4-positive T cells in the nerve fascicles. The polyneuropathy was responsive to steroid therapy. Analyses of serological human leukocyte antigen (HLA) types indicated that none of the patients possessed a high-risk HLA type known to be associated with adult T-cell leukemia (ATL), whereas they did have high responsive alleles to HTLV-I env similar to that observed in HAM. Nucleotide sequence analysis of the HTLV-I tax region demonstrated the B subgroup in all patients. This study suggests that HTLV-I infection can result in the development of a familial form of polyneuropathy that is associated with distinct HLA class I alleles, which might possibly involve a distinct virus subtype. PMID- 16036799 TI - Nectin-1/HveC Mediates herpes simplex virus type 1 entry into primary human sensory neurons and fibroblasts. AB - Immunocytochemistry detects nectin-1/HveC, nectin-2/HveB, and HVEM/HveA on the surface of sensory neurons and fibroblasts grown as primary cultures from human dorsal root ganglia. Viral entry into these cultured cells was assayed by infection with a recombinant herpes simplex virus type 1 (HSV-1) expressing green fluorescent protein. Soluble, truncated nectin-1 polypeptide, as well as polyclonal and monoclonal antibodies against nectin-1, inhibited infection of neurons, whereas polypeptides and antibodies capable of inhibiting HSV-1 interaction with nectin-2 and herpesvirs entry mediator (HVEM) failed to prevent infection of neuronal cells. These results demonstrate that nectin-1 is the primary receptor for HSV-1 entry into human fetal neurons. Viral entry into fibroblasts was also reduced by soluble nectin-1 but not by soluble HVEM. However, in contrast to the results obtained with neurons, antibodies against receptors failed to inhibit entry into fibroblasts, indicating that unlike neurons, fibroblasts have multiple receptors or mechanisms for HSV-1 entry. PMID- 16036800 TI - Macrophage chemoattractant protein-1 levels in cerebrospinal fluid correlate with containment of JC virus and prognosis of acquired immunodeficiency syndrome- associated progressive multifocal leukoencephalopathy. AB - In the highly active antiretroviral therapy (HAART) era, the role of the inflammatory response in acquired immunodeficiency syndrome (AIDS)-related progressive multifocal leukoencephalopathy (PML) remains controversial. In this study, JC virus DNA load and levels of cytokines were determined in cerebrospinal fluid (CSF) from 32 human immunodeficiency virus (HIV)-1-infected patients with confirmed PML who underwent HAART; cytokines were also measured in 12 HIV positive controls. Predictors of survival were analyzed by Cox's models. Macrophage chemoattractant protein (MCP)-1 levels were significantly higher in PML patients than in controls (mean +/- SD, 2.45 +/- 0.64 versus 1.32 +/- 0.64 log(10) pg/ml, P<.0001). In PML patients, the higher concentration of MCP-1 correlated with lower JC viral load (r=-.405, P=.036). Higher concentrations of MCP-1 in CSF were associated with longer survival on HAART after adjusting for CD4 counts (for each log(10) pg/ml higher, hazard ratio for death 0.28, 95% confidence interval 0.08--1.00). Predictors of shorter survival were lower baseline CD4 counts, higher JCV DNA concentrations, lower Karnofsky, and no prior HAART exposure. These results showed that higher CSF levels of MCP-1, an inflammatory cytokine, were correlated with better prognosis in HAART-treated patients with PML. PMID- 16036801 TI - Recurrent transverse myelitis following neurobrucellosis: immunologic features and beneficial response to immunosuppression. AB - The authors report the clinical course and immune system response of a patient with disease-associated recurrent transverse myelitis (TM) following cerebral infection with Brucellosis melitensis. The patient suffered four recurrences of his TM (each at a distinct spinal cord level) over the course of 2 years following his initial presentation, which ultimately progressed to quadriplegia. He had progressively declining cerebrospinal fluid (CSF) brucella antibody titers, suggesting a postinfectious, rather than an infectious, etiology. The authors simultaneously examined the expression of multiple cytokines in the CSF of this patient using cytokine antibody arrays and found a marked elevation of interleukin (IL)-6, IL-8, and macrophage chemoattractant protein (MCP)-1 levels relative to controls. Quantitative enzyme-linked immunosorbent assay (ELISA) analysis of the CSF confirmed a 1700-fold elevation of IL-6 and more modest elevations of IL-8 and MCP-1. IL-6 levels returned to baseline following treatment of the patient with intravenous cyclophosphamide and plasma exchange and the patient experienced a significant and sustained recovery of function. PMID- 16036802 TI - MSRV/HERV-W/syncytin and its linkage to multiple sclerosis: the usability and the hazard of a human endogenous retrovirus. PMID- 16036803 TI - Poliovirus type 1 infection of murine PRNP-knockout neuronal cells. AB - Transfection of the prion protein gene (Prnp) into prion-deficient mouse cells was shown to reduce the replication of coxsackievirus B3, an enterovirus. Because mice can be susceptible to poliovirus infection by parenteral routes, the authors tested the susceptibility to poliovirus-1 (PV-1) of a panel of murine neuronal cell lines differing in their ability to express Prnp. The investigated cell lines (prionless HpL3.4 cells, HpL3.4 cells transfected with a Prnp vector, HpL3.4 cells transfected with a void vector, wild-type Hw3.5 Prnp(+/+) cells) expressed the murine homologue (Tage4) of human poliovirus receptor (CD155/hPVR). PV-1 infection of Prnp(-/-) HpL3.4 cells resulted in the production of high viral titers, though viral antigens could be detected in only 0.5% to 2% of cells. Wild type Prnp(+/+) cells and prionless cells transfected with the Prnp gene were not permissive to PV-1. Results of viral titration and immunofluorescence were confirmed by conventional polymerase chain reaction (PCR) and quantitative real time PCR. Exposure to PV-1 had no influence on the gene expression profile of Prnp(+/+) cells. In contrast, PV-1 infection was associated with upregulation of several genes in permissive Prnp(-/-) cell cultures: type I interferon (IFN) genes, IFN-related developmental regulator 1 (IFNRD1), tumor necrosis factor superfamily member 13b (TNFSF13b), interleukin (IL) - 7, granulocyte/macrophage colony-stimulating factors (CSFs), hepatocyte growth factor (HGF), vascular endothelial growth factor-A, transforming growth factors beta1 and beta3 (TGFb1, TGFb3), as well as a variety of bone morphogenetic proteins endowed with neuroprotective activity. Distinction of permissive from nonpermissive neuronal cells on the basis of Prnp expression suggests that prion-deficient mice could represent an extraordinarily sensitive animal model for poliovirus infection. PMID- 16036804 TI - Tumor necrosis factor alpha leads to increased cell surface expression of CXCR4 in SK-N-MC cells. AB - Both host and viral factors play an important role in the pathogenesis of human immunodeficiency virus (HIV)-associated bran injury. In this study, the authors examined the interactions between tumor necrosis factor (TNF)-alpha, CXCR4, the alpha chemokine receptor, and three HIV isolates, including the T-tropic viruses, HIV-1(MN) and HIV-1(IIIB), and the dual tropic virus, HIV-1(89.6). The authors show by flow cytometry that treatment of differentiated SK-N-MC cells with TNF alpha induces a significant increase in the cell surface expression of CXCR4 in a time- and dose-dependent manner. The effect is partly regulated at the level of transcription. To assess the biological significance of this finding, we show that TNF-alpha potentiates the ability of the above mentioned HIV isolates to induce neuronal apoptosis and that the effect is significantly reduced by pretreating cells with monoclonal antibodies to either CXCR4 and TNF-alpha. Together these results suggest that TNF-alpha may render neuronal cells vulnerable to the apoptotic effects of HIV by increasing the cell surface expression of CXCR4 and thus identify another mechanism by which TNF-alpha contributes to the pathogenesis of HIV-associated brain injury. PMID- 16036805 TI - Neuronal activity regulates viral replication of herpes simplex virus type 1 in the nervous system. AB - Herpes simplex virus types 1 and 2 (HSV-1, -2) infect and also establish latency in neurons. In the present study, the authors investigated the influence of neuronal activity on the replication of HSV-1. The results showed that the sodium channel blocker tetrodotoxin (TTX) and the inhibitory neurotransmitter gamma aminobutyric acid (GABA) could significantly increase viral replication in primary neuronal cultures, by two- to fourfold. In contrast, KCl reduced viral production by at least 80% in the same cultures. Inhibitors of GABA(A) receptors completely abolished the effects of GABA. Intravitreously injected TTX in a mouse corneal scarification model enhanced the viral titers > 10-fold in both the trigeminal ganglia and the brain. At 2 h post infection, both TTX and GABA significantly up-regulated the levels of transcription for the viral immediate early (IE) genes ICP0, ICP4, and ICP27, as revealed by real time PCR. These results indicate that the neuronal excitation status may dictate the efficiency of HSV-1 viral replication, probably by regulating the levels of viral IE gene expression. These are the first findings connecting neuronal activity to the molecular mechanisms of HSV replication in the nervous system, which may significantly influence our view of herpesvirus infection and latency. PMID- 16036806 TI - Prevalence and risk factors for human immunodeficiency virus-associated neurocognitive impairment, 1996 to 2002: results from an urban observational cohort. AB - To assess prevalence and risk factors for human immunodeficiency virus (HIV) related neurocognitive impairment (NCI), the authors performed a 7-year survey in the period 1996 to 2002. A total of 432 patients were examined. HIV-related NCI was diagnosed in 238 patients (55.1%), meeting the HIV dementia (HIV-D) criteria in 45 (10.4%). The prevalence of both NCI and HIV-D did not change significantly during the study period. Compared with patients without NCI, patients with NCI were older (40.4 versus 38.2 years; P = .003), had a higher prevalence of positive HCV serology (61.1% versus 38.9%; P = .003), and a lower nadir CD4 cell count (156 versus 222 cells/microl; P < .001). Compared with patients seen during 1996 to 1999, patients with NCI seen during 2000 to 2002 were older (40.7 versus 38.8 years; P = .004), had a less advanced disease stage (previous acquired immunodeficiency syndrome [AIDS] 28.8% versus 65.7%; P < .001) and a higher nadir CD4 count (174 versus 132 cells/microl; P = .026). This study showed an unchanged prevalence of both HIV-related NCI and HIV-D in the period 1996 to 2002. The authors found evidences for new additional potential risk factors for HIV-related NCI (older age, lower nadir CD4 count, positive hepatitis C virus [HCV] serology), and for a change of risk factors for NCI in the late highly active antiretroviral therapy (HAART) era (older age, less advanced disease, higher nadir CD4 count). PMID- 16036807 TI - T cell-mediated restriction of intracerebral murine cytomegalovirus infection displays dependence upon perforin but not interferon-gamma. AB - The authors have previously reported that adoptive transfer of splenocytes suppresses murine cytomegalovirus (MCMV) brain infection following intracerebroventricular injection of immunodeficient mice and that depletion of Thy 1.2+ T lymphocytes abolishes this suppressive effect. Here the authors report that splenocytes depleted of CD4+ T lymphocytes prior to adoptive transfer retained their ability to control viral expression in the brain. In sharp contrast, depletion of the CD8+ T-cell population prior to transfer abolished the suppressive effect, with sixfold greater expression in the brain than when undepleted splenocytes were used. The authors went on to examine the contributions of cytokine- and perforin-mediated mechanisms in controlling MCMV brain infection using splenocytes from major histocompatibility (MHC)-matched IFN gamma -knockout (GKO), and perforin-knockout (PKO) mice. When used in adoptive transfer studies, splenocytes from GKO mice controlled viral expression; however, cells from PKO mice could not control reporter gene expression or viral DNA replication in brain tissues. The authors have previously reported that the levels of the T-cell chemoattractant CXCL10 are highly elevated in the brains of MCMV-infected mice. Here the authors found that the receptor for this ligand, CXCR3, was not essential in mediating the suppressive effects of adoptive transfer. These data indicate that peripheral CD8+ T cells control MCMV brain infection through a perforin-mediated mechanism and that neither IFN-gamma nor CXCR3 play a critical role in this neuroprotective response. PMID- 16036808 TI - Cocaine increases human immunodeficiency virus type 1 neuroinvasion through remodeling brain microvascular endothelial cells. AB - Cocaine is a suspected cofactor in human immunodeficiency virus (HIV)-associated dementia but cocaine's effects are not clear. Herein the authors describe investigations of the mechanisms by which cocaine increases HIV-1 invasion through brain microvascular endothelial cells (BMVECs). Cocaine binds to a site on BMVECs, which is not a biogenic amine transporter, a binding site for estrogen, or a muscarinic receptor and for which benztropine and tamoxifen have the highest affinity. Cocaine treatment of BMVECs disrupts intercellular junctions and induces cell ruffling, which could account for their increased permeability and decreased electrical resistance. HIV-1 enters BMVECs by macropinocytosis and is transported to lysosomes and inactivated. In cocaine treated BMVECs, the virus enters and persists in large cytoplasmic "lakes." Cocaine exposure of BMVECs up-regulates transcription of genes important in cytoskeleton organization, signal transduction, cell swelling, vesicular trafficking, and cell adhesion. The toxicity of cocaine for the blood-brain barrier may lead to increased virus neuroinvasion and neurovascular complications of cocaine abuse. PMID- 16036809 TI - Diffusion tensor imaging of subcortical brain injury in patients infected with human immunodeficiency virus. AB - Diffusion tensor imaging (DTI) was used to derive in vivo tissue status measurements of subcortical brain regions that are vulnerable to injury in human immunodeficiency virus (HIV)-infected patients. Quantitative measurements, including the mean diffusivity (MD) and fractional anisotropy (FA), were determined in lateralized basal ganglia (caudate and putamen) and centrum semiovale in 11 well-characterized HIV patients and in 11 control subjects. DTI measurements were examined for patterns of relationship with markers of clinical and cognitive progression. DTI measures acquired in subcortical regions were significantly correlated with loss of function in specific cognitive domains. Significant relationships were identified between measures for putamen and verbal memory (MD), visual memory (FA), working memory (FA), and overall cognitive impairment (MD). Measures for caudate (FA) were significantly correlated with visual memory. Measures for centrum semiovale were significantly correlated with visual memory deficits (MD) and visuoconstruction (FA). Relationships between anisotropy measures and anemia (basal ganglia) and CD4 counts (centrum semiovale) were also observed. Findings from this investigation indicate that DTI is a sensitive tool for correlating neuroanatomic pathologic features with specific cognitive deficits in patients with HIV infection. PMID- 16036810 TI - Gene expression profiles in peripheral blood mononuclear cells from patients with subacute sclerosing panencephalitis using oligonucleotide microarrays. AB - To investigate the molecular basis for measles virus persistence in patients with subacute sclerosing panencephalitis (SSPE), the authors used a high-density oligonucleotide microarray, and found that the expression of granulysin in peripheral blood mononuclear cells was significantly lower in the patients than in the controls. By a quantitative reverse transcriptase-polymerase chain reaction, the mRNA levels of granulysin were decreased in 30 SSPE patients, and were increased in 7 measles patients, as compared to the 23 controls. These results imply that granulysin might play a role in the host defense against measles virus and possibly be involved in the pathogenesis or pathophysiology of SSPE. PMID- 16036811 TI - Histone deacetylase inhibitors induce reactivation of herpes simplex virus type 1 in a latency-associated transcript-independent manner in neuronal cells. AB - Histone acetylation is implicated in the regulation of herpes simplex virus type 1 (HSV-1) latency. However, the role of histone acetylation in HSV-1 reactivation is less clear. In this study, the well-established model system, quiescently infected, neuronally differentiated PC12 (QIF-PC12) cells, was used to address the participation of histone acetylation in HSV-1 reactivation. In this model, sodium butyrate and trichostatin A (TSA), two histone deacetylase inhibitors, stimulated production of infectious HSV-1 progeny from a quiescent state. To identify viral genes responsive to TSA, the authors analyzed representative alpha, beta, and gamma viral genes using quantitative real-time polymerase chain reaction. Only the latency-associated transcript (LAT) accumulated in response to TSA treatment, under culture conditions that restricted virus replication and spread. This led the authors to evaluate the importance of LAT expression on TSA induced reactivation. In QIF-PC12 cells, the LAT deletion mutant virus dLAT2903 reactivated equivalently with its wild-type parental strain (McKrae) after TSA treatment, as well as forskolin and heat stress treatment. Both viruses also reactivated equivalently from latently infected trigeminal ganglia explants from rabbits. In contrast, there was a marked reduction in the recovery of dLAT2903, as compared to wild-type virus, from the eyes of latently infected rabbits following epinephrine iontophoresis. These combined in vitro, ex vivo, and in vivo data suggest that LAT is not required for reactivation from latently infected neuronal cells per se, but may enhance processes that allow for the arrival of virus at, or close to, the site of original inoculation (i.e., recrudescence). PMID- 16036812 TI - The bane of surrogate decision-making: defining the best interests of never competent persons. PMID- 16036813 TI - A model for validating an expert's opinion in medical negligence cases. PMID- 16036814 TI - A comparison of Cruzan and Schiavo: the burden of proof, due process, and autonomy in the persistently vegetative patient. PMID- 16036815 TI - Conflict of duty: capital punishment regulation and AMA medical ethics. PMID- 16036816 TI - Cord blood storage: property and liability issues. PMID- 16036820 TI - Errors of omission in the treatment of prehospital chest pain patients. AB - OBJECTIVE: Despite the widespread use of standard treatment protocols, there are few published data regarding paramedic protocol adherence. In this descriptive study, the authors sought to assess the frequency and nature of deviations from a standardized treatment protocol for the chief complaint of chest pain. They also sought to quantify any time delays in treatment of potential ischemic cardiac chest pain. METHODS: A retrospective review of written documentation obtained from four ambulance services in a mid-Atlantic state was completed. A convenience sample of consecutive emergency medical services (EMS) records was obtained from January 2001 to May 2002, and 75 calls were selected from each service (N = 300). RESULTS: Neither the median scene times nor the response times varied among the four services in the study. However, the suburban ambulance service (service 1) did have a significantly longer transport time (19 minutes) than the rural (14 minutes) and the urban (11 and 10 minutes) services (p < 0.05). Documentation of history and physical characteristics varied widely for each service. The patient took aspirin 10% of the time prior to EMS arrival, yet paramedics gave it additionally 50% of the time, while nitroglycerin was given in 73% of cases of suspected cardiac ischemia. Posttreatment vital signs for nitroglycerin were documented 30% of the time for three of the four services, while the other service documented these 75% of the time. Medical command contact varied by agency (80-100%), as did the receipt and completion of medical orders. CONCLUSIONS: Paramedics may delay transport of patients with potential cardiac ischemia. Deviations from protocol occur frequently and the care documented for prehospital patients with chest pain is variable. The expected care described by written protocols does not correlate with the treatment documented. PMID- 16036821 TI - Noninvasive fireground assessment of carboxyhemoglobin levels in firefighters. AB - OBJECTIVES: Carboxyhemoglobin (COHb) levels can be estimated by chemical analysis of exhaled alveolar breath. Such noninvasive measurement could be used on the fireground to screen both firefighters (FFs) and victims. The purpose of this study was to assess the feasibility of using a hand-held carbon monoxide (CO) monitoring device to screen for CO toxicity in FFs under field conditions. METHODS: Informed consent was obtained from all participants. Using a hand-held breath CO detection device, COHb readings were collected at baseline, and then as FFs exited burning buildings after performing interior fire attack and overhaul with self-contained breathing apparatus (SCBA) during live-fire training. Ambient CO levels were occasionally measured in interior areas where the FFs were working to assess the degree of CO exposure. RESULTS: Baseline COHb readings of 64 FFs ranged from 0% to 3% (mean 1%, median 1%). One hundred eighty-four COHb readings were collected during training exercises. The mean and median COHb levels were 1%. The maximum value in a FF wearing SCBA was 3%; values of 14%, 5%, and 4% were measured in instructors who were not properly wearing SCBA. Ambient CO readings during fire attack ranged from 75 to 1,290 ppm, and the ambient CO reading for overhaul ranged from 0 to 130 ppm. When the device was used for interior CO monitoring, washout time limited its utility for COHb monitoring in FFs. CONCLUSIONS: A hand-held CO monitoring device adapted for estimation of COHb levels by exhaled breath analysis can feasibly be deployed on the fireground to assess CO exposure in FFs. PMID- 16036822 TI - Measuring the EMS patient access time interval and the impact of responding to high-rise buildings. AB - OBJECTIVES: To measure the patient access time interval and characterize its contribution to the total emergency medical services (EMS) response time interval; to compare the patient access time intervals for patients located three or more floors above ground with those less than three floors above or below ground, and specifically in the apartment subgroup; and to identify barriers that significantly impede EMS access to patients in high-rise apartments. METHODS: An observational study of all patients treated by an emergency medical technician paramedics (EMT-P) crew was conducted using a trained independent observer to collect time intervals and identify potential barriers to access. RESULTS: Of 118 observed calls, 25 (21%) originated from patients three or more floors above ground. The overall median and 90th percentile (95% confidence interval) patient access time intervals were 1.61 (1.27, 1.91) and 3.47 (3.08, 4.05) minutes, respectively. The median interval was 2.73 (2.22, 3.03) minutes among calls from patients located three or more stories above ground compared with 1.25 (1.07, 1.55) minutes among those at lower levels. The patient access time interval represented 23.5% of the total EMS response time interval among calls originating less than three floors above or below ground and 32.2% of those located three or more stories above ground. The most frequently encountered barriers to access included security code entry requirements, lack of directional signs, and inability to fit the stretcher into the elevator. CONCLUSIONS: The patient access time interval is significantly long and represents a substantial component of the total EMS response time interval, especially among ambulance calls originating three or more floors above ground. A number of barriers appear to contribute to delayed paramedic access. PMID- 16036823 TI - Calling emergency medical services for acute stroke: a study of 9-1-1 tapes. AB - OBJECTIVE: To obtain a better understanding of how stroke events are communicated to 9-1-1 telecommunicators, and how telecommunicators and emergency medical services (EMS) personnel respond to such calls. METHODS: The authors identified 104 patients with a hospital discharge diagnosis of stroke or transient ischemic attack who were transported to hospital by ambulance in two North Carolina counties during 1999 and 2000. Ambulance call reports were abstracted and linked to 9-1-1 call center audiotapes, which were transcribed and verified. RESULTS: Of the 104 calls, 44 were made by medical personnel, 38 by a family member, eight by a bystander or neighbor, five undetermined, and three by other nonmedical personnel. In only six instances (6%) was the call placed by the patient. The most common symptoms reported were altered mental status (40%), trouble walking (32%), impaired speech (27%), and abnormal breathing (27%). Although the word "stroke" was often used (45%), 9-1-1 telecommunicators classified the calls as a stroke in only 31% of cases. However, in the majority of cases (79%), paramedics were dispatched at the highest priority. The median time from dispatch of EMS to patient arrival at the hospital was 41 minutes, approximately half of which was spent at the scene. CONCLUSION: Although typical stroke symptoms are commonly described, calls are often not classified as "strokes" by telecommunicators. Nevertheless, because of the symptoms reported during the calls, the majority of cases are treated as high priority by telecommunicators. PMID- 16036824 TI - Validation of using EMS dispatch codes to identify low-acuity patients. AB - OBJECTIVE: To validate the predictive ability of previously derived emergency medical services (EMS) dispatch codes to identify patients with low-acuity illnesses. METHODS: This prospective descriptive study was conducted in Rochester, New York. An expert panel reviewed and modified a previously derived set of low-priority EMS dispatch codes. Patients assigned these 21 codes between July 2002 and June 2003 were included for further analysis. Dispatch data and level of EMS care were recorded for each dispatch code. The proportion of low acuity patients (i.e., those who received only basic life support (BLS) care or those who were not transported using lights and sirens) was determined using previously established definitions. Codes were defined as associated with low acuity patients if the lower bound of the 95% confidence interval (CI) exceeded 90%. Medical records for patients identified as high-acuity were reviewed to evaluate whether the advanced life support (ALS) level care that was provided had a clinical impact. RESULTS: Emergency medical services cared for 43,602 patients during the study, and 7,540 were dispatched as low-priority. We found that 7,197 (95%; 95% CI: 95-96%) of these patients met low-acuity criteria and that 11 of the evaluated codes were validated, with low-acuity care provided at least 90% of the time. Of the 343 patients identified as high-acuity, 62 (18%; 95% CI: 14-23%) were determined to have received interventions that had a clinical impact. CONCLUSIONS: This study prospectively validates 11 EMS dispatch codes as being associated with low-acuity patients. These codes could be used to triage EMS patients based on dispatch information. PMID- 16036825 TI - Prehospital pain management: a comparison of providers' perceptions and practices. AB - OBJECTIVE: To assess the knowledge of emergency medical technicians-paramedics (EMT-Ps) and compare their practice perceptions with actual pain management interventions in adults and pediatric patients (adolescents and children) with chest pain (CP), extremity injuries, or burns. METHODS: This study included a cross-sectional survey of EMT-Ps and review of the emergency medical services (EMS) system patient care database. EMT-Ps were surveyed for: 1) knowledge of pain treatment protocol; 2) estimated number of CP, extremity injury, or burn encounters and the frequency of morphine administration; and 3) barriers to providing morphine. Data on patients transported with any above conditions and those who received morphine were abstracted from the EMS patient care database. Data were analyzed using descriptive statistics, and 95% confidence intervals (CIs) were calculated. RESULTS: Of 202 EMT-Ps, 155 (77%) completed the survey. Eighty-two percent reported knowledge of pain treatment protocol for both adults and pediatric patients. For adults, EMT-Ps estimated they administered morphine to 37% with CP (95% CI 35, 40), 24% with extremity injuries (95% CI 17, 30), and 89% with burns (95% CI 52, 99). In children and adolescents, inability to assess pain (93%) was the most common reason for withholding morphine. According to the EMS database, 5% of adults with CP (95% CI 4, 5), 12% extremity injuries (95% CI 8, 15), and 14% burns (95% CI 8, 20) received morphine. In children and adolescents, 3% with extremity injuries (95% CI 1, 5) and 9% with burns (95% CI 0, 26) received morphine. Pain score was documented in 67.0% of adult patients, compared with only 4.0% in pediatric patients (Delta = 63.0%, 95% CI: 60, 65). CONCLUSIONS: Significant disparity exists between EMT-Ps' perceptions of acute pain assessment and the frequency of providing analgesia and their actual practice. Children and adolescents had less documentation of pain assessment and received less analgesic interventions compared with adults. Inability to assess pain may be an important barrier to the provision of analgesia. PMID- 16036826 TI - Prehospital pain management in children suffering traumatic injury. AB - Prehospital pain management has become an important emergency medical services (EMS) patient care issue. OBJECTIVES: To describe the frequency of EMS and emergency department (ED) analgesic administration to injured children; to describe factors associated with the administration of analgesia by EMS; and to assess whether children with lower-extremity fractures receive analgesia as frequently as do adults with similar injuries. METHODS: This was a retrospective study of children (age < 21 years) who were transported by EMS between January 2000 and June 2002 and had a final hospital diagnosis of extremity fractures or burns. Secondarily, children with lower-extremity fractures were compared with a cohort of EMS-transported adults with similar injuries and transported during the same study period. Receipt of and time of parenteral analgesia were recorded. RESULTS: Seventy-three children met the inclusion criteria. The mean (range) age of this sample was 12.4 (0.9-21) years, with only four patients aged < 5 years. A majority of the patients were male (49/73, 67.1%) and sustained femur (20/73, 27.4%) or tibia/fibula (26/73, 35.6%) fractures. Few pediatric patients received prehospital analgesia (16/73, 21.9%), while a majority received analgesia in the ED (58/73, 79.4%). Prehospital analgesia was associated with earlier patient treatment than that administered in the ED (22.3 +/- 5.9 min vs. 88.3 +/- 38.2 min). Comparing children (n = 33) with adults (n = 76) with similar lower extremity fractures, a small insignificant difference was found in the rate of prehospital analgesia between children and adults (7/33, 21.2%, vs. 20/56, 26.3%). CONCLUSION: Few pediatric patients receive prehospital analgesia, although most ultimately received ED analgesia. Few factors were identified that could be associated with EMS oligoanalgesia. No difference was found between children and adults in the rates of EMS analgesia. PMID- 16036827 TI - Discontinuation of droperidol for the control of acutely agitated out-of-hospital patients. AB - OBJECTIVE: To identify the effects of the removal of droperidol as a treatment option for sedation of agitated out-of-hospital patients. METHODS: This was a retrospective review conducted January 1, 2001, through December 5, 2002, of patients with an out-of-hospital diagnosis of agitation who received either droperidol or midazolam prior to arrival in the emergency department (ED). The need for continuous cardiac or pulse oximetry monitoring, intubation, critical care ED management, intensive care unit admission, and mortality was reviewed. RESULTS: Seventy-one patients received droperidol or midazolam for acute agitation in the out-of-hospital setting. Forty-one patients received droperidol in 2001 (D2001); three patients received midazolam in 2001 (M2001). No patients received droperidol in 2002, and 27 patients received midazolam (M2002). Comparing the D2001 and M2002 groups, the need for continuous pulse oximetry monitoring in the ED [14/41 (34.1%) versus 18/27 (66.7%)], intubations [4/41 (9.8%) versus 10/27 (37.0%)], critical emergency medical services transports [5/41 (12.2%) versus 11/27 (40.7%)], critical ED care cases [6/41 (14.6%) versus 11/27 (40.7%)], and intensive care unit admissions [6/13 (46.2%) versus 14/15 (93.3%)] were increased in the M2002 group. No difference was found in the frequencies of ED cardiac monitoring, hospital admission, complications, or death. CONCLUSIONS: Since the removal of droperidol as a treatment option for out of-hospital agitated patients, the authors have observed an increased frequency of continuous pulse oximetry monitoring, intubation, ED critical care management, and intensive care unit admission in patients requiring chemical sedation for control of agitation in the out-of-hospital setting. PMID- 16036828 TI - Association of heat index and patient volume at a mass gathering event. AB - In 1999, a department of emergency medicine was asked to provide medical care at a football stadium with a capacity of 61,625. Over four seasons, the department's experience has been that the number of patients seen during a game correlates closely with game-time heat and humidity (heat index). OBJECTIVE: To determine how closely the heat index is associated with the number of patients who will require care at a mass gathering event. METHODS: This was a retrospective review of all patient care from 1999 to 2003 at a Division I college football stadium located in the southeastern United States. All patrons seen in two emergency care centers (ECCs) were included. To control for stadium attendance, the Pearson product-moment correlation (PPMC) was calculated for each game. This statistical tool determines whether there is a positive correlation between heat index and ratio of number of patients cared for per 10,000 patrons. RESULTS: A total of 20 games occurred, and the heat index ranged from 33 to 92. Number of patients varied from 15 to 74, and stadium attendance ranged from 53,371 to 61,625. The PPMC was calculated as 0.607, which indicates a strong positive correlation between heat index and patient volume (p < 0.005). Linear modeling predicts that for every 10-degree increase in the heat index, three more patients per 10,000 patrons will require care. CONCLUSIONS: In this retrospective study, the heat index was strongly associated with the volume of patients who would be seen at a mass gathering event. PMID- 16036829 TI - The effect of a voice assist manikin (VAM) system on CPR quality among prehospital providers. AB - Numerous studies have documented poor cardiopulmonary resuscitation (CPR) performance among prehospital providers during both simulated and actual resuscitations. Previous studies have shown that a real-time, voice assist manikin (VAM) system may improve CPR performance. OBJECTIVE: To determine whether VAM prompting would improve CPR performance by prehospital providers during simulated resuscitation. METHODS: In this prospective, randomized, crossover design, 114 prehospital providers performed two 3-minute sessions of one-rescuer CPR on a VAM-resuscitation manikin: one round with the VAM feature turned on and one with the feature turned off. The primary outcomes were measured at 15-second intervals and included the fraction of correct compressions, the mean compression depth, the fraction of correct ventilations, and the mean ventilation tidal volume. Generalized estimating equations were used to analyze the repeated measures. RESULTS: The VAM prompting was not directly associated with correct compressions during one-rescuer CPR in a cohort of subjects naive to the system. However, the general decay in correct compressions seen over 3 minutes was attenuated with VAM prompting. Neither the compression depth nor the decay in compression depth over time was affected by VAM prompting. In contrast, VAM prompting did affect the fraction of correct ventilations and attenuated the time dependent decline in correct ventilations in tidal volume. CONCLUSIONS: Use of VAM did not directly improve compression or ventilation rate or quality in this cohort of prehospital providers. However, use of VAM did prevent decay of compression and ventilation performance over time. PMID- 16036830 TI - The impact of a new CPR assist device on rate of return of spontaneous circulation in out-of-hospital cardiac arrest. AB - OBJECTIVE: The San Francisco Fire Department deployed an automated, load distributing-band chest compression device (AutoPulse, Revivant Corporation) to evaluate its function in a large urban emergency medical services (EMS) service. A retrospective chart review was undertaken to determine whether the AutoPulse had altered short-term patient outcome, specifically, return of spontaneous circulation (ROSC). METHODS: AutoPulse cardiopulmonary resuscitation (A-CPR) was used by paramedic captains responding to adult cardiac arrests with an average +/ SD response time of 15 +/- 5 minutes. The primary endpoint was patient arrival to an emergency department with measurable spontaneous pulses. The manual CPR comparison group was case-matched for age, gender, initial presenting electrocardiogram rhythm, and the number of doses of Advanced Cardiac Life Support medications as a proxy for treatment time. Matching was performed by an investigator blinded to outcome and treatment group. RESULTS: Sixty-nine AutoPulse uses were matched to 93 manual-CPR-only cases. A-CPR showed improvement in the primary outcome when compared with manual CPR with any presenting rhythm (A-CPR 39%, manual 29%, p = 0.003). When patients were classified by first presenting rhythm, shockable rhythms showed no difference in outcome (A-CPR 44%, manual 50%, p = 0.340). Outcome was improved with A-CPR in initial presenting asystole and approached significance with pulseless electrical activity (PEA)(asystole: A-CPR 37%, manual 22%, p = 0.008; PEA: A-CPR 38%, manual 23%, p = 0.079). CONCLUSION: The AutoPulse may improve the overall likelihood of sustained ROSC and may particularly benefit patients with nonshockable rhythms. A prospective randomized trial comparing the AutoPulse with manual CPR in the setting of out-of-hospital sudden cardiac arrest is under way. PMID- 16036831 TI - Near-continuous, noninvasive blood pressure monitoring in the out-of-hospital setting. AB - OBJECTIVES: This study was conducted to test out-of-hospital performance of a noninvasive radial artery tonometry device to assess blood pressure (BP), providing readings every 10-12 seconds. The primary objective was to determine the correlation between noninvasive BPs calculated with radial artery tonometry and standard oscillometric cuff methods. The secondary objective was to determine whether the difference observed between the two techniques was consistent over the range of BPs measured. METHODS: This prospective trial enrolled adults transported by helicopter (n = 9 patients), fixed-wing airplane (n = 1), or ground vehicle (n = 10) of a single transport service. Patients had BP assessed simultaneously, by both standard automatic cuff and radial artery tonometry device, every 5 minutes. Data were assessed with correlation coefficients, and Bland-Altman techniques were utilized to assess for bias over the range of mean arterial pressures (MAPs) encountered. For all tests, p was set at 0.05. RESULTS: No major problem with radial artery tonometry device field performance was noted. There were 139 pairs of MAP assessments in 20 patients. The correlation coefficient for the two assessment modalities was 0.96. Bland-Altman bias plot and Pitman's test (p = 0.11) revealed good correlation between the two assessment mechanisms over the entire range of MAPs (42 to 163 mm Hg) encountered in the study. CONCLUSION: The radial artery tonometry device provided MAP assessments that were highly correlated with readings from a standard oscillometric device. The radial artery tonometry device performed well in a variety of patient types and in multiple transport vehicles, and there was no sign that its performance was adversely affected by the out-of-hospital setting. PMID- 16036832 TI - Feasibility of sternal intraosseous access by emergency medical technician students. AB - OBJECTIVE: Emergency medical technician-basic (EMT-B) providers are not trained to establish vascular or intraosseous (IO) access on critically ill patients. This study was conducted to examine the feasibility of training EMT-B students to correctly place a commercial sternal IO infusion device (FAST-1). METHODS: Twenty nine EMT-B students attended a two-hour training session. Subjects were subsequently tested in FAST-1 application using a modified resuscitation mannequin permitting IO needle deployment. Two observers assessed correct IO application and technique. Results were analyzed using descriptive statistics (binomial proportions and medians with 95% confidence intervals). Inter-rater agreement of observations was evaluated using kappa statistics and intraclass correlation coefficients (ICCs). RESULTS: Inter-rater agreement ranged from fair to excellent (kappa = 0.37-1.00) for all parameters except sternal notch identification (kappa = -0.03). Reliabilities of elapsed times were good (ICC = 0.83, 0.31). Correct identification of the sternal notch was accomplished by 28 of 29 students (96.6%; 95% CI: 82.2-99.9%). Correct application of the IO target patch was achieved by 29 of 29 (100.0%; 88.1-100.0%). First-attempt successful IO needle deployment was achieved by 16 of 29 (55.2%; 35.7-73.6%). Overall successful IO needle deployment within four attempts was achieved by 27 of 29 (93.1%; 77.2-99.2%). The protective dome was correctly applied by 27 of 29 (93.1%; 77.2-99.2%). The median time to needle deployment was 27.5 seconds (95% CI: 24-31). The median time to dome placement was 50 seconds (95% CI: 42-55). CONCLUSIONS: EMT-B students with minimal training demonstrated limited success with applying a commercial sternal IO device. Clinical application by EMT-Bs on critically ill patients may be possible with more intensive training. PMID- 16036833 TI - HEMS vs. Ground-BLS care in traumatic cardiac arrest. AB - OBJECTIVE: To assess whether a top-level type of prehospital care, made of helicopter, physician, and advanced life support (ALS) procedures, improves the outcome of blunt trauma victims found in cardiac arrest (CA) as compared with a simpler type, composed of ground ambulance, nurse, and expanded basic life support (BLS). METHODS: This was a cohort study from the data set of a prospective, population-based, 12-month study targeting the 1,200,000 inhabitants of the Italian region Friuli Venezia Giulia. RESULTS: Fifty-six victims received the higher level of care (helicopter emergency medical services [HEMS] group) and 73 received the lower one (ground-BLS group). The two groups were homogeneous for mechanism of injury, gender, and time interval before cardiopulmonary resuscitation (CPR). Age was lower in the ground-BLS group. The percentage of patients in which CPR was attempted was significantly higher in the HEMS group (43% vs. 20%; CI 0.061 to 0.379). On-scene return of spontaneous circulation (ROSC) was also more likely in the HEMS group (37.5% of attempted CPRs vs. 6.6%; CI 0.027 to 0.591). None of the patients evacuated from the scene without ROSC ever attained it in hospital. This policy was virtually exclusive to the ground BLS group. Survival to hospital discharge was 3.5% (severely disabled) in the HEMS group and 0% in the ground-BLS group (CI -0.008 to 0.078). CONCLUSION: A top level type of prehospital care had significantly more chances to resuscitate blunt trauma victims found in CA as compared with a simpler level. No significant benefit on long-term outcome was found, but more cases might be needed in future studies because of the inevitably low number of survivors. PMID- 16036834 TI - Ketamine chemical restraint to facilitate rescue of a combative "jumper". PMID- 16036835 TI - Transport of the ventricular assist device-supported patient: a case series. PMID- 16036838 TI - An algorithmic approach to prehospital airway management. AB - Airway management, including endotracheal intubation, is considered one of the most important aspects of prehospital medical care. This concept paper proposes a systematic algorithm for performing prehospital airway management. The algorithm may be valuable as a tool for ensuring patient safety and reducing errors as well as for training rescuers in airway management. PMID- 16036839 TI - Defining the learning curve for paramedic student endotracheal intubation. AB - BACKGROUND: Proficiency in endotracheal intubation (ETI) is assumed to improve primarily with accumulated experience on live patients. While the National Standard Paramedic Curriculum recommends that paramedic students (PSs) perform at least five live ETIs, these training opportunities are limited. OBJECTIVE: To evaluate the effects of cumulative live ETI experience, elapsed duration of training, and clinical setting on PS ETI proficiency. METHODS: The authors used longitudinal, multicenter data from 60 paramedic training programs over a two year period. The PSs reported outcomes (success/failure) for all live ETIs attempted in the operating room (OR), the emergency department (ED), the intensive care unit (ICU), and other hospital or prehospital settings. Fixed effects logistic regression was used to model up to 30 consecutive ETI efforts by each PS, accounting for per-PS clustering. For each patient, the authors evaluated the association between ETI success and the PS's cumulative number of ETIs, adjusted for clinical setting, elapsed number of days from the first ETI encounter, and the interaction (cumulative ETIs x elapsed days). Predicted probability plots were constructed depicting the "learning curve" overall and for each clinical setting. Results. Between one and 74 ETIs (median 7; IQR 4-12) were performed by each of 802 PSs. Of 7,635 ETIs, 6,464 (87.4%) were successful. Stratified by clinical setting, 6,311 (82.7%) ETIs were performed in the OR, 271 (3.6%) in the ED, 64 (0.8%) in the ICU, 86 (1.1%) in other in-hospital settings, and 903 (11.8%) in the prehospital setting. For the 7,398 ETIs included in the multivariate analysis, cumulative number of ETI was associated with increased adjusted odds of ETI success (odds ratio 1.067 per ETI; 95% CI: 1.044-1.091). ETI learning curves were steepest for the ICU and prehospital settings but lower than for other clinical settings. CONCLUSIONS: Paramedic student ETI success improves with accumulated live experience but appears to vary across different clinical settings. Strategies for PS airway education must consider the volume of live ETIs as well as the clinical settings used for ETI training. PMID- 16036840 TI - Populations at risk for intubation nonattempt and failure in the prehospital setting. AB - OBJECTIVES: Pediatric cardiac arrest patients and adult traumatic arrest patients are perceived as more difficult to endotracheally intubate than adult cardiac arrest patients. The study hypothesis was that these populations were at higher risk of endotracheal intubation failure compared with adult cardiac arrest patients and that paramedics would more frequently defer attempts to intubate these patients. METHODS: This was a retrospective, observational study analyzing oral endotracheal intubations on pediatric cardiac arrest, adult traumatic arrest, and adult cardiac arrest patients over 66 months. Homogeneity of intubation nonattempt and endotracheal intubation failure was studied with chi square analysis. Relative risks (RRs) with 95% confidence intervals (CIs) were used to compare pediatric cardiac arrest with adult traumatic arrest with adult cardiac arrest nonattempt rates and endotracheal intubation failure rates. RESULTS: 2,669 oral endotracheal intubations were included. There was a significant difference in intubation nonattempts and intubation failure between the combined pediatric cardiac arrest and adult traumatic arrest groups and the adult cardiac arrest cohort (RR 7.24, 95% CI 5.73, 9.16 for nonattempt; RR = 2.33, 95% CI 1.93, 2.83 for intubation failure). Both groups individually showed significant risk for intubation nonattempt and endotracheal intubation failure compared with adult cardiac arrest, with the pediatric cohort at higher risk for failure and the adult traumatic arrest cohort at higher risk for nonattempt. CONCLUSIONS: There was significant risk of intubation nonattempt and intubation failure in the pediatric cardiac arrest and adult traumatic arrest cohorts compared with the adult cardiac arrest population, with the pediatric cohort being at particularly high risk for intubation failure and the adult traumatic arrest cohort at higher risk for nonattempt. PMID- 16036841 TI - Reliability of paramedic ratings of laryngoscopic views during endotracheal intubation. AB - BACKGROUND: Prior studies have related prehospital endotracheal intubation (ETI) difficulty to paramedic visualization of the vocal cords using the Cormack-Lehane (C-L) scale. However, the reliability of paramedic C-L ratings has not been formally studied. OBJECTIVE: To evaluate the reliability of C-L and a more recently described scale, percentage of glottic opening (POGO), when used by paramedics to rate laryngoscopic views during ETI. METHODS: Twenty-five standard slide images of laryngoscopic views were obtained during ETI. The 25 images were duplicated to facilitate evaluation of intrarater agreement (total 50 slides). Seven paramedics rated the degree of vocal cord visualization in each image using C-L (I-IV, ordinal scale; I = full visualization of vocal cords, IV = only epiglottis seen) and POGO (0-100 continuous scale; 0 = no vocal cords seen, 100 = full visualization of vocal cords). We assessed intra- and interrater reliabilities using Cohen's multirater kappa for C-L and intraclass correlation coefficients (ICCs) for POGO. RESULTS: C-L showed variable intrarater reliability (kappa range = 0.37-0.90) and poor interrater reliability (Cohen's multirater kappa = 0.22). POGO demonstrated good to excellent intrarater reliability (one way random-effects ICC range = 0.57-0.87) and fair to good interrater reliability (two-way random-effects ICC = 0.59, 95% Confidence interval: 0.48-0.71). CONCLUSIONS: Paramedic C-L ratings exhibit poor intra- and interrater reliabilities. Paramedic POGO ratings exhibit fair to good intra- and interrater reliabilities. POGO may be more appropriate than C-L for prehospital clinical and scientific application. Reliability must be formally evaluated for any proposed laryngoscopic exposure classification system. PMID- 16036842 TI - Endotracheal tube placement by EMT-Basics in a rural EMS system. AB - OBJECTIVE: To evaluate the effectiveness of an intubation-training module and special-waiver project in which Emergency Medical Technician (EMT)-Basics were trained to perform endotracheal intubations in a rural community. METHODS: This was a prospective observational study over a four-year period (July 1998 through May 2002) of all intubation attempts by EMT-Basics in the field. The authors observed intubation data, training methods, and quality-assurance methods of a special-waiver project agreed to by the State Department of Public Health to train and allow EMT-Basics to intubate patients. Data were from documentation unique to the project. Project documentation evaluated the placement and complication(s) of endotracheal tube (ETT) placement after arrival to the emergency department. An intubation attempt was defined as direct laryngoscopy. A successful attempt was defined as an appropriately sized ETT placed and secured in the trachea below the vocal cords and above the carina. Confirmation of placement in the field included accepted clinical methods and the use of qualitative colorimetric end-tidal carbon dioxide detectors. The EMT-Basics were trained using a paramedic curriculum, including operating room intubations on live adult patients. All patients were in either cardiopulmonary or respiratory arrest. RESULTS: Thirty-two intubations were performed by EMT-Basics. Thirty attempts were successful and two were unsuccessful (94%; 95% confidence interval [CI] 80-98%). Unsuccessful ETT placements were managed with accepted basic life support airway standards. There were no unrecognized esophageal ETT placements (0%; 95% CI 0-11%). CONCLUSION: This study demonstrated that with an intensive training program using selected highly motivated providers and close monitoring, a program of EMT-Basic ETT placement in a rural setting can achieve acceptable success rates in patients in cardiac or respiratory arrest. PMID- 16036843 TI - Evolution of statewide EMS drug formularies and regulations. AB - OBJECTIVE: To characterize and follow the variability present in statewide emergency medical services (EMS) medication formularies across the United States over a ten-year period. METHODS: Investigators contacted the lead EMS agencies in all 50 states during three years (1992, 1997, and 2002). Using a standardized form, the investigators collected information about each state's prehospital medication policies, including whether a statewide EMS medication formulary existed, the authority of local medical directors to modify it, and what medications it contained. The investigators then sorted states into categories based on the regulatory intent of their EMS medication policies and compared medication listings across years. RESULTS: Responses were obtained from all 50 states (n = 50, 100%) during each of the survey periods. There appeared to be a trend toward stricter state control and toward less variation between statewide formularies. State regulations in seven states stopped allowing local medical directors to retain full control of their systems' formularies, and eight states implemented mandatory statewide formularies. There was a trend toward more consistency between states, with more "most commonly" listed medications (6.9% in 1992 versus 22.1% in 2002) and fewer "least commonly" listed medications (58.3% in 1992 versus 42.3% in 2002). Controversial medications such as neuromuscular blockers and thrombolytics appeared in a small but increasing number of statewide formularies. CONCLUSIONS: Considerable variation was found among statewide EMS medication formularies, both in how they were established and in their contents. Although several states continued to rely solely on local medical direction, there seemed to be a trend toward more uniformity and stricter state control over prehospital medication formularies during the study period. PMID- 16036844 TI - Presentation and survival of prehospital apparent sudden infant death syndrome. AB - BACKGROUND: Prehospital providers are often involved in the resuscitation of apparent sudden infant death syndrome (SIDS) victims; however, data are few on the presentation and outcome of these patients. OBJECTIVES: To describe the presentation and determine the survival rate of infants who have an unwitnessed, prehospital arrest consistent with SIDS (apparent SIDS), and to compare the presentation of infants with a final diagnosis of SIDS with those who presented as apparent SIDS but had a different final diagnosis. METHODS: This was a secondary analysis of data from a controlled trial whose methodology has been previously described. The setting was two large, urban emergency medical services (EMS) systems of Los Angeles and Orange Counties, California. The population included 113 apparent SIDS victims from the original interventional study who had a prehospital, unwitnessed arrest consistent with SIDS, defined by the scenario of an infant aged =12 months being placed to sleep and later found in full arrest (pulseless and apneic). Data collected included ethnicity, gender, arrest etiology, signs of death (lividity, rigor mortis), prehospital interventions, return of spontaneous circulation (ROSC), arrest rhythm, code 3 transport (lights and sirens), and survival to hospital discharge. RESULTS: One hundred ten of 113 apparent SIDS patients had survival data; 0 of 110 (95% CI 0% to 3.3%) survived, although ROSC was achieved in 5%; for three patients data on survival were missing. Arrest rhythms were determined in 94% of the subjects: asystole 87%, pulseless electrical activity (PEA) 8%, and ventricular fibrillation 4%. Only 50 of 113 (44%) of the EMS records documented code 3 transport; the remainder of the records were ambiguous. SIDS was the final coroner's diagnosis for 79 of 113 (70%) of the cases. Other causes of death in these apparent SIDS victims included respiratory causes (12%), asphyxiation (3%), abuse (2%), congenital heart disease (2%), sepsis (2%), other (4%), and unknown (5%). Apparent SIDS victims with a final diagnosis of SIDS were more likely to show signs of death (27/79, 34% vs. 5/34, 15%, p = 0.035) and were less likely to have a rhythm of PEA (4/77, 5% vs. 5/31, 16%, p = 0.08), although the latter result was not statistically significant. CONCLUSIONS: Apparent SIDS victims have a dismal prognosis; all infants presenting with apparent SIDS died, even the 30% whose final diagnosis was not SIDS. Given that there were no survivors, new prehospital policies are needed governing the use of lights and sirens, resuscitation decisions including termination of resuscitation, provision of grief support to families, and incident stress debriefing for prehospital personnel. PMID- 16036845 TI - Ten years of police defibrillation: program characteristics and personnel attitudes. AB - OBJECTIVE: In 1992,a study to evaluate the effectiveness of police defibrillation was initiated in seven suburban police departments. This paper describes the characteristics of those programs and the attitudes of chiefs and officers ten years after the commencement of that study. METHODS: A 32-item survey instrument including questions on department demographics, defibrillation program characteristics, and attitudes was mailed to the chiefs. A separate 25-item survey was developed to assess officers' attitudes; a non-management officer distributed and collected these surveys anonymously. Responses are reported descriptively. RESULTS: All seven chiefs and 78% of the officers completed surveys. Police arrive prior to emergency medical services on 80% or more of calls, with a mean +/- standard deviation response interval of 3.1 +/-1.7 minutes. All departments identify a program coordinator and six a medical director; three departments have a quality-improvement program. Five departments have liability coverage for the use of an automated external defibrillator (AED), and no department reported any liability-related problems. Five of the chiefs and 82% of the officers disagreed or strongly disagreed that liability issues are a concern or a source of hesitation in using the AED. Ninety-six percent of the officers agreed or strongly agreed that police AED use is beneficial to cardiac arrest victims. All seven police chiefs and 89% of the officers agreed or strongly agreed that AED use by police is appropriate. CONCLUSIONS: In these seven police departments with ten years of program experience, defibrillation has become integrated into the law-enforcement culture. Liability issues are not a prominent concern of chiefs or officers. Police chiefs and officers in these departments continue to support police-based defibrillation programs. PMID- 16036846 TI - The safety and efficacy of prehospital needle and tube thoracostomy by aeromedical personnel. AB - BACKGROUND: Aeromedical crews routinely use needle thoracostomy (NT) and tube thoracostomy (TT) to treat major trauma victims (MTVs) with potential tension pneumothorax; however, the efficacy of prehospital NT and TT is unclear. OBJECTIVES: To explore the efficacy of aeromedical NT and TT in MTVs. METHODS: A retrospective chart review was performed using prehospital medical records and the county trauma registry over a seven-year period. All MTVs undergoing placement of NT or TT by aeromedical personnel were included; patients with incomplete data were excluded. Descriptive statistics were used to report the incidence of air release, clinical improvement (improved breath sounds or compliance if intubated, decreased dyspnea if nonintubated), and vital signs improvements (systolic blood pressure [SBP] increase to =90 mm Hg or increase by 5 mm Hg if < 90 mm Hg; heart rate improvement to 60-100 beats/min, increase by 10 beats/min if < 60 BPM, or decrease by 10 beats/min if > 100 beats/min; oxygen saturation increase if < 95%) for both NT and TT as documented in prehospital medical records. Survival and improvement in SBP based on trauma registry data were recorded for patients stratified by initial SBP. RESULTS: A total of 136 procedures (89 NTs and 47 TTs) in 81 patients were identified using prehospital medical records over a four-year period. Response rates to NT (60% overall, 32% vital signs) and TT (75% overall, 60% vital signs) were high. Vital signs improvements were observed more often in patients with a pulse and in nonintubated patients. A total of 168 patients were identified in the trauma registry over the seven-year study period. Normalization of SBP was observed in two-thirds of patients with a field SBP = 90 mm Hg and one-third of patients in whom field SBP could not be obtained. A small but significant proportion of patients undergoing prehospital NT and TT, including some with prehospital hypotension and high injury severity, survived to hospital discharge. The incidence of complications was low. CONCLUSIONS: Aeromedical crews appear to appropriately select MTVs to undergo field NT or TT. A low incidence of complications and a small but significant group of unexpected survivors support continued use of this procedure by aeromedical personnel. PMID- 16036847 TI - EMS and emergency department physician triage: injury severity in trauma patients transported by helicopter. AB - BACKGROUND: Many trauma patients who are not severely injured arrive at trauma centers via helicopter emergency medical services (HEMS). OBJECTIVE: To compare the injury severity of patients sent to trauma centers by HEMS from community emergency departments (EDs) with the injury severity of those triaged by prehospital providers to HEMS directly from accident scenes. METHODS: All records were reviewed from trauma-related missions during 1997for a single HEMS system, extracting information on location, time of day, patient age and gender, mechanism of injury, initial vital signs, Revised Trauma Score (RTS), and the extent of care required during transport. These records were then matched with outcome information routinely supplied to the HEMS system by affiliated trauma centers. Information from patients flown directly from scenes was then compared with that for patients flown from community EDs. RESULTS: Information was obtained for 658 patients flown from scenes and 345 flown from community EDs. There were similar proportions of patients in the two groups, with Injury Severity Scale (ISS) scores less than 6 (11.0% vs. 13.5%), between 6 and 14 (47.0% vs. 49.3%), and greater than 15 (42.0% vs. 37.1%); these were not statistically different (p > 0.05). There was also no significant difference between the groups in the RTS, mean ISS score, intensive care unit length of stay, hospital length of stay, or disposition. CONCLUSIONS: Scene and interhospital HEMS trauma missions in this system involve patients of similar injury severities. Prehospital providers may triage trauma patients to HEMS transport with proficiency similar to that of community ED physicians. PMID- 16036848 TI - The state of EMS education research project: characteristics of EMS educators. AB - OBJECTIVES: The preparation of emergency medicine practitioners occurs at a variety of levels, via individuals with various levels of experience and training, and through a variety of oversight organizations. The purpose of this study was to quantify the characteristics of those recognized as prehospital emergency medical services (EMS) educators, the type and amount of infrastructure available to facilitate the learning process, and what attributes and common practices the profession values. METHODS: The respondents (n = 1,691) were generated randomly from a nationwide systematic sample of all known EMS instructors. The sample was adequate to generalize to the 15,000 EMS educators. Face validity and content validity were assured through use of a 24-member focus group of EMS educators, practitioners, and administrators, who reviewed the questionnaire. Reliability estimates were generated via Cronbach's alpha and Kuder-Richardson 20 and 21 and ranged from 0.59 to 0.83, with a grand mean for the seven study constructs of 0.70. Factor analysis with varimax rotation explained 40% to 66% of the respective construct variance, with a grand mean of 56% of the response variance explained. RESULTS: The respondents were satisfied with their teaching experience (98%), expected to continue (80%), and taught because of a perceived dearth of qualified instructors. Only half of the respondents utilized federally generated curricular materials, and approximately 20% were uncomfortable in assessing psychomotor skills. The relationship of testing to assessment and skill performance appeared to be the most significant pedagogic challenge. CONCLUSIONS: The respondents were a senior group of educators committed to the training of new providers who will comprise the foundation of future EMS educational efforts at the national level. PMID- 16036849 TI - Use of treat-and-release medical directives for paramedics at a mass gathering. AB - INTRODUCTION: Paramedics provide a substantial proportion of care at mass gatherings but do not typically release patients without physician assessment. OBJECTIVE: To evaluate treat-and-release medical directives implemented at a large single-day summer rock concert. METHODS: Medical directives allowed paramedics to administer acetaminophen, dimenhydrinate, diphenhydramine, or polymyxin B ointment for common complaints without evidence of serious illness on history or examination. After treatment, patients were released or transferred to a medical facility according to predefined criteria. Patient demographics, chief complaint, treatment, and disposition were obtained from paramedic records. To determine whether any patients released by paramedic subsequently required ambulance transport, all ambulance records were searched for a period of eight hours before to 24 hours after the event. RESULTS: More than 450,000 people attended the concert, with 1,870 presenting for medical attention. Four hundred seven patients received medications under the directives. No disposition was recorded in 13 cases. Two hundred ninety-nine patients were treated with acetaminophen, of whom 269 (90.0%) were released and 23 (7.7%) required additional care. Sixty-two patients received dimenhydrinate, 44 (71%) were released, and 14 (23%) required transport. Thirty-six patients received diphenhydramine, and 34 (94%) were released. Ten patients received polymyxin B and were released. No patient released by paramedics was found to have later required ambulance transport. CONCLUSIONS: Treat-and-release medical directives for paramedics at mass gatherings may help divert patients from requiring care at a medical facility. Future research is needed to determine the safety (morbidity and mortality) of these directives. PMID- 16036850 TI - Emergency medical services in Connecticut. AB - This article describes emergency medical services (EMS) systems in Connecticut, beginning with a historical perspective. The discussion of statewide oversight of the EMS system includes legislative and regulatory mandates as well as recent external reviews of the system. Medical oversight of EMS care and services is provided by sponsor hospitals rather than individual medical directors. Most of the 169 cities and towns in the State maintain or contract for local EMS, and have traditionally resisted regionalization. This snapshot of the EMS system in Connecticut can serve as a reference for comparison of EMS systems in other jurisdictions. PMID- 16036851 TI - Effectiveness of a simple Internet-based disaster triage educational tool directed toward Latin-American EMS providers. AB - BACKGROUND: A previous survey demonstrated a lack of standardization related to disaster triage among Latin-American providers. OBJECTIVE: To assess the effectiveness of a short Internet-based educational intervention in disaster and mass-casualty triage. Using three Spanish Internet emergency medical services (EMS) forums, Latin-American providers were invited to participate in the study. The tool consisted of two educational modules: an introduction to disaster triage module and a START (simple triage and rapid treatment) module. Pre- and post intervention tests were administered, each consisting of five standardized scenarios. Factorial analysis was used to measure the weight of each scenario. The first and fifth scenarios were identical for intraclass correlation. Skill retention was assessed through a one-month follow-up survey. Statistical analysis was performed using chi-square and Fisher's exact test. A total of 55 EMS providers participated in the study. Five of 55 (9.1%) participants correctly answered four or more scenarios on the pretest intervention, compared with 53 of 55 (96.4%) on the posttest [p < 0.001, relative risk 10.60 (95% CI 4.59-24.49)]. Similar findings were obtained for those accurately triaging all five scenarios, with zero of 55 (0%) in the pretest compared with 49 of 55 in the posttest (p < 0.001). Follow-up at one month was 69%. Four or more scenarios were correctly answered at follow-up by 34 of 38 (89.5%) respondents. No significant difference was noted compared with the immediate post-course survey (p = 0.18). Although initial ability of the cohort to accurately triage patients was suboptimal, a short Internet-based educational tool significantly impacted the cohort's ability to perform triage in a simulated patient environment. This improvement was maintained after one month. PMID- 16036852 TI - Pulseless electrical activity: a diagnostic challenge for EMS. PMID- 16036853 TI - Blood-borne pathogens among firefighters and emergency medical technicians. AB - OBJECTIVE: Firefighters and emergency medical services (EMS) personnel have the potential for occupational exposures to blood, which increases their risk for occupational blood-borne infection. To address this concern, the authors conducted a literature review of occupational blood exposures, the seroprevalence of blood-borne pathogens among these workers, and the seroprevalence of blood borne pathogens among the patients they serve. METHODS: A MEDLINE search was conducted, and all identified articles that described surveys of exposures to blood or surveillance of blood-borne infections among firefighters and/or emergency medical technicians (EMTs) in the United States were reviewed. For hepatitis B, only seroprevalence surveys conducted after the 1992 requirement by the Bloodborne Pathogens Standard to offer vaccination to potentially exposed employees were included. RESULTS: From these data, the expected number of annual occupational hapatitis C virus seroconversions was estimated to be between 5.8 and 118.9 per 100,000 employee-years for EMT-paramedics, between 3.4 and 33.7 per 100,000 for firefighter-EMTs, and up to 3.6 per 100,000 for firefighters (non EMT). CONCLUSIONS: This review suggests there are a limited number of studies addressing this issue, and these studies have numerous limitations. Despite the expected occupational seroconversions and recognizing the limitations in drawing conclusions from these studies, it appears that firefighters and EMS personnel do not have an elevated seroprevalence of hepatitis C virus compared with the general population. Improved exposure surveillance programs would clarify exposure risks and identify potential interventions for firefighters and EMS personnel. PMID- 16036856 TI - Expanded emergency medical services: the failure of an experimental community health program. PMID- 16036858 TI - Transcriptomic analysis of F344 rat nasal epithelium suggests that the lack of carcinogenic response to glutaraldehyde is due to its greater toxicity compared to formaldehyde. AB - Formaldehyde is cytotoxic and carcinogenic to the rat nasal respiratory epithelium inducing tumors after 12 months. Glutaraldehyde is also cytotoxic but is not carcinogenic to nasal epithelium even after 24 months. Both aldehydes induce similar acute and subchronic histopathology that is characterized by inflammation, hyperplasia, and squamous metaplasia. Because early aldehyde induced lesions are microscopically similar, we investigated whether transcriptional patterns using cDNA technology could explain the different cancer outcomes. Treatments included 1-, 5-, or 28-day exposure by nasal instillation of formaldehyde solution (400 mM) or glutaraldehyde solution (20 mM). Animals were euthanized and the nasal respiratory epithelium removed for gene expression analysis and a subset of rats treated for 28 days was processed for microscopic examination. RNA was isolated and processed for expression assessment using Clontech Atlas Toxicology II Arrays. Both aldehydes induced hyperplasia, squamous metaplasia, and inflammatory infiltrates with scattered apoptotic bodies in the epithelium covering luminal surfaces of the nasoturbinate, maxilloturbinate, and nasal septum. A subset of 80 genes that were the most variant between the treated and control included the functional categories of DNA repair and apoptosis. Hierarchical clustering discriminated chemical treatment effects after 5 days of exposure, with 6 clusters of genes distinguishing formaldehyde from glutaraldehyde. These data suggest that although both aldehydes induced similar short-term cellular phenotypes, gene expression could distinguish glutaraldehyde from formaldehyde. The gene expression patterns suggest that glutaraldehyde's lack of carcinogenicity may be due to its greater toxicity from lack of DNA repair, greater mitochondrial damage, and increased apoptosis. PMID- 16036859 TI - Profiles of metabolites and gene expression in rats with chemically induced hepatic necrosis. AB - This study investigated whether integrated analysis of transcriptomics and metabolomics data increased the sensitivity of detection and provided new insight in the mechanisms of hepatotoxicity. Metabolite levels in plasma or urine were analyzed in relation to changes in hepatic gene expression in rats that received bromobenzene to induce acute hepatic centrilobular necrosis. Bromobenzene-induced lesions were only observed after treatment with the highest of 3 dose levels. Multivariate statistical analysis showed that metabolite profiles of blood plasma were largely different from controls when the rats were treated with bromobenzene, also at doses that did not elicit histopathological changes. Changes in levels of genes and metabolites were related to the degree of necrosis, providing putative novel markers of hepatotoxicity. Levels of endogenous metabolites like alanine, lactate, tyrosine and dimethylglycine differed in plasma from treated and control rats. The metabolite profiles of urine were found to be reflective of the exposure levels. This integrated analysis of hepatic transcriptomics and plasma metabolomics was able to more sensitively detect changes related to hepatotoxicity and discover novel markers. The relation between gene expression and metabolite levels was explored and additional insight in the role of various biological pathways in bromobenzene induced hepatic necrosis was obtained, including the involvement of apoptosis and changes in glycolysis and amino acid metabolism. The complete Table 2 is available as a supplemental file online at http://taylorandfrancis.metapress.com/openurlasp?genre=journal&issn=0192-6233. To access the file, click on the issue link for 33(4), then select this article. A download option appears at the bottom of this abstract. In order to access the full article online, you must either have an individual subscription or a member subscription accessed through www.toxpath.org. PMID- 16036860 TI - An improved model of isolated hemoperfused porcine livers using pneumatically driven pulsating blood pumps. AB - Existing liver perfusion models are largely limited by high degrees of ischemic and reperfusion injury and the lack of standardization. To establish a highly standardized perfusion model and minimize reperfusion injury, a porcine liver perfusion model was developed using an artificial heart pump (Buecherl Artificial Heart). This model is characterized by pneumatically driven and pressure controlled blood pumps with pulsating flow characteristics. The perfusion parameters and the integrity of the perfused organ were assessed using hemodynamic and hepatic function tests. In eight porcine liver perfusion experiments the system allowed maintaining stable and physiologic organ function over 3 hours by bile production (5.5 +/- 3.1 ml/30 minutes, resp. 22.9 +/- 8.4 ml cumulative at 180 minutes), oxygen consumption (2.2 +/- 0.2 ml/min/100 g overall mean) and significantly better liver enzyme levels (AST 19.5 +/- 10.1 U/l/100 g, ALT 2.1 +/- 0.8 U/l/min, LDH 57.8 +/- 24.2 U/l/100 g) compared to previous studies. It was also possible to reduce the circulating blood volume to 1,000 ml and to create a compact perfusion system that is adoptable to other organ systems such as the kidneys. The compact size and the absence of magnetic components also allow a use for advanced imaging techniques. In conclusion this optimized perfusion system provides a sound basis for future studies in the area of hepatotoxicity and pharmacology. PMID- 16036861 TI - Skeletal pathology in white storks (Ciconia ciconia) associated with heavy metal contamination in southwestern Spain. AB - In 1998, a mine tailings dyke in southwestern Spain broke, flooding the Agrio Guadiamar river system with acid tailings up to the borders of one of the largest breeding colonies of white storks in the western Palearctic, Dehesa de Abajo. Over the following years, a high proportion of nestlings developed leg defects, prompting this study. Ten fledglings with leg deformities from the spill area were compared with 11 normal storks of the same year class from another region far from the spill. However, metals were analyzed as a continuum rather than by site, as reference birds also contained high levels of metals. Gross pathology of the legs was supported by histopathology, which showed that bone remodeling activity was greater in the deformed storks, which also had more irregular subperiosteal bone, and tended to have higher residual islets of cartilage in their metaphyses, which, in turn were related to metal contaminant residues. Both Ca and P in bone were affected independently by metals. Deformed birds had lower serum bone alkaline phosphatase. Bone malformations, measured by leg asymmetry, was only partially explained by bone metals, indicating that a combination of factors was involved with the abnormal development in these young storks. PMID- 16036862 TI - Cartilage dysplasia and tissue mineralization in the rat following administration of a FGF receptor tyrosine kinase inhibitor. AB - PD176067 is a reversible and selective inhibitor of fibroblast growth factor receptor tyrosine kinase, and was in preclinical development as an angiogenesis inhibitor for the treatment of solid tumors. A 14-day oral toxicity study of PD176067 in young female rats (7 weeks old) was conducted at doses of 2.5, 5, and 10 mg/kg/day (15, 30, and 60 mg/m(2), respectively). Skeletal changes, and vascular and soft tissue mineralization were observed as primary drug-related toxicities. To determine if these changes are specific to young, rapidly growing animals with increased vascular and osseous development, PD176067 was administered to mature (11 months old) rats. Female rats received PD176067 by gavage for 14 days at doses of 2.5, 5, and 10 mg/kg/day and necropsied on day 15. Clinical signs of toxicity were seen at > or =5 mg/kg and one death occurred at 10 mg/kg. Physeal dysplasia (distal femur, proximal tibia, sternum) occurred in all drug-treated animals and was characterized by dose-related increased thickness of the zones of chondrocyte proliferation and hypertrophy, and marked thickening of the zone of ossification. Cartilage hyperplasia was characterized by proliferation of chondrocytes along margins of the synchondrosis and subperiosteum of sternebrae. Serum phosphorus levels increased 47% and 166% at 5 and 10 mg/kg, respectively. Mineralization of cardiac myocytes, aorta, various arteries, renal tubules, and gastric mucosa and muscularis was seen at 10 mg/kg, and consistent with the presence of calcium-phosphorus deposition. Physeal changes occurred at similar plasma PD176067 exposures in young and mature rats (AUC > or = 4.83 microg.hr/mL). PD176067 produced morphologically similar lesions in young and adult rats. PMID- 16036863 TI - PTHrP and PTH/PTHrP receptor 1 expression in odontogenic cells of normal and HHM model rat incisors. AB - Parathyroid hormone related peptide (PTHrP) was discovered as a causative factor of humoral hypercalcemia of malignancy (HHM). We examined PTHrP and its receptor (PTHR1) expression patterns in odontogenic cells in normal and HHM model rat incisors. Nontreated nude rats serving as the normal control and HHM model rats produced by implantation of PTHrP-expressing tumor (LC-6) cells were prepared. HHM rats fractured its incisor, and histopathologically, restrict population of odontoblasts showed findings classified as "shortening of high columnar odontoblasts" and "dentin niche." The incisors were immunostained against PTHrP and PTHR1. In normal rats, PTHrP and PTHR1 colocalized in ameloblasts, cementoblasts, and odontoblastic cells from mesenchymal cells to columnar odontoblasts. In high columnar odontoblasts, PTHrP solely expressed. In the HHM animals, although the expression patterns were identical to those of the normal rats in normal area, the shortened high columnar odontoblasts maintained PTHR1 expression and dentin niche comprising odontoblastic cells expressed both proteins. In the HHM model, the protein expression patterns changed in the odontoblastic cells with histological anomalies, and thus direct relations between the anomalies and PTHrP/PTHR1 axis are suggested. PMID- 16036864 TI - A study of the potential of the pig as a model for the vaginal irritancy of benzalkonium chloride in comparison to the nonirritant microbicide PHI-443 and the spermicide vanadocene dithiocarbamate. AB - A porcine model was established to test the mucosal toxicity potential of a thiophene thiourea (PHI-443)-based anti-HIV microbicide and a vanadocene-based spermicide, vanadocene dithiocarbamate (VDDTC) in comparison to benzalkonium chloride (BZK). Nine domestic pigs (Duroc) in nonestrus stage received a single intravaginal application of 2% BZK, 2% PHI-443, or 0.1% VDDTC-containing gel. At various times after gel application, cell differentials and levels of inflammatory cytokines (IL-1beta, IL-4, IL-6, IL-8, IL-10, IL-18, IFN-gamma, and TNF-alpha) in cervicovaginal lavage (CVL) fluid were monitored by flow cytometry and ELISA, respectively. Eight pigs were exposed intravaginally to a gel with and without BZK or VDDTC for 4 consecutive days and vaginal tissues were scored histologically for inflammation using a new scoring system. Only CVL fluid from pigs exposed to BZK showed a significant increase of IL-1beta, IL-8, and also IL 18 production when compared to the controls, PHI-443 or VDDTC-treated groups. Maximum levels of BZK-induced IL-1beta (100-fold), IL-8 (2,500-fold), IL-18 (80 fold), and IFN-gamma (10-fold) were found at 24 hours. In the in vivo porcine vaginal irritation model, increased levels of vaginal IL-1beta, IL-8, and IL-18 were associated with histological changes consistent with vaginal inflammation. These results demonstrate that key cervicovaginal inflammatory cytokines are useful in vivo biomarkers for predicting the mucosal toxicity potential of vaginal products in the physiologically relevant and sensitive porcine model. PMID- 16036865 TI - Incidences of selected lesions in control female Harlan Sprague-Dawley rats from two-year studies performed by the National Toxicology Program. AB - The NTP has a long history of using Fischer rats and has compiled a large database of incidences of lesions seen in control animals. Such a database is lacking for Harlan Sprague-Dawley (SD) rats. The intention of this paper is to report spontaneous lesions observed in female vehicle control Harlan SD rats, and to compare the incidence in 2 strains of rats (Fischer and Harlan SD) used in NTP studies. Female Harlan SD rats served as the test animals for a special series of 2-year studies. Male rats were not used in these studies. Complete histopathology was performed on all animals, and the pathology results underwent comprehensive NTP pathology peer review. The most commonly observed neoplasms in these female control Harlan SD rats were mammary gland fibroadenoma (71%), tumors of the pars distalis of the pituitary (41%) and thyroid gland C-cell tumors (30%). Female Fischer rats had incidences of 44% for mammary gland fibroadenomas, 34% for tumors of the pars distalis, and 16% for thyroid gland C-cell tumors. Fischer rats had a 15% incidence of clitoral gland tumors, while the Harlan SD rats had an incidence of < 1%. In contrast to Fischer F344 rats, the Harlan SD rats had a high incidence of squamous metaplasia of the uterus (44.2%). Squamous metaplasia is not a lesion commonly observed in NTP control Fischer rats. The Harlan SD rats had a very low incidence of mononuclear cell leukemia (0.5%), compared with an incidence of 24% in female Fischer rats. PMID- 16036866 TI - Anatomical localization of cartilage degradation markers in a surgically induced rat osteoarthritis model. AB - Osteoarthritis (OA) is a degenerative disease characterized by an irreversible loss of articular cartilage. Although surgically induced animal OA models are commonly used in drug efficacy assessment, degradation of type II collagen, an important component of articular cartilage is not routinely evaluated. Here, the medial meniscectomy surgical model (MMT) in Lewis rats was evaluated for proteoglycan loss with toluidine blue staining and collagen degradation with immunohistochemical staining for a collagen cleavage C-neoepitope, using a novel anti-type II collagen neoepitope antigen (TIINE) antibody. Femorotibial joints were collected for histology at 0 (no surgery), 3, 7, 14, 21, 28, 35, and 42 days postsurgery. Following MMT surgery, the medial tibial articular cartilage had proteoglycan matrix loss by day 3 that reached subchondral bone by days 28-42. Femoral cartilage damage occurred by day 14. TIINE staining was present at basal levels in growth plates and articular cartilage of all joints while all MMT treated animals had increased intensity and area of staining in erosions that colocalized with proteoglycan loss. The MMT model produces a progressive pattern of cartilage damage resembling human OA lesions, making it useful, when evaluated with cartilage biomarkers, for assessing changes in cartilage degradation. PMID- 16036867 TI - The comparative power of the discriminant methods used in toxicological pathology. AB - It is highly desirable to use experimental methodologies in toxicological pathology that combine statistical power, practicality, and objective reviewability to detect small differences. The different ways of gathering data at the microscope can result in clear differences in power to discriminate small, but real, differences between treated and control rodent groups with nonneoplastic lesions. Six alternative methods of gathering and analysing results are compared. They are referred to as the Measuring, Ordering, Scoring (or Grading), Pair-contrast, Outside-control, and Affected methods. Measuring and Ordering methods are uniformly more powerful than other more common and highly esteemed methods, such as Scoring/Grading. From the practical perspective, Measuring and Ordering can be applied objectively, reviewed objectively, and interpreted to standards that are widely accepted as valid throughout experimental science e.g., using confidence limits and intervals. They also are intuitively natural extensions of routine toxicological histopathological examinations. Establishing a small difference between control and treated groups is commonly a problem when reporting no-observed-effect levels. Ordering is the recommended method for assessing if a small difference between treated and control groups is within chance variation or is the result of a true treatment effect, when measurement is impractical. PMID- 16036868 TI - Preclinical safety and pharmacokinetics of recombinant human factor XIII. AB - Factor XIII (FXIII) is a thrombin-activated protransglutaminase responsible for fibrin clot stabilization and longevity. Deficiency in FXIII is associated with diffuse bleeding and wound-healing disorders in humans. This report summarizes results from several studies conducted in adult cynomolgus monkeys (M. fascicularis) to evaluate the safety and pharmacokinetics of recombinant human factor XIII A(2) dimer (rFXIII). Intravenous slow bolus injection of rFXIII resulted in the expected formation of the heterotetramer rA(2)cnB(2), prolonged circulating half-life (5-7 days), and increased plasma transglutaminase activity. Recombinant FXIII was well tolerated as a single dose up to 20 mg/kg rFXIII (2840 U/kg), as repeated daily doses up to 6 mg/kg (852 U/kg) for 14 days, and as 3 repeated doses of 8 mg/kg (1136 U/kg) separated by 14 days. Overt toxicity occurred after a single intravenous injection of = 22.5 mg/kg rFXIII (3150 U/kg), or with 2 doses of = 12.5 mg/kg (1775 U/kg) administered within 72 hours. The rFXIII-mediated toxicity was expressed as an acute systemic occlusive coagulopathy. Evaluation of plasma samples from dosed animals demonstrated formation of cross-linked fibrin/fibrinogen oligomers and higher-order protein aggregates, which are hypothesized to be responsible for the observed vessel occlusion and associated embolic sequelae. These results demonstrate that rFXIII mediated toxicity results from exaggerated pharmacological activity of the molecule at supraphysiological concentrations. The absence of observed toxicological effect with repeated intravenous doses up to 8 mg/kg (1136 U/kg) was used to support an initial clinical dose range of 0.014 to 0.35 mg/kg (2-50 U/kg). PMID- 16036869 TI - Fluorouracil plus leucovorin induces submandibular salivary gland enlargement in rats. AB - The administration of 5-fluorouracil (FU) and leucovorin (LV) to rats induced a previously unreported sialoadenitis-like toxicity. Four different treatment regimens were used: daily-times-5 iv or ip injections of LV (200 mg/kg) followed 30 minutes later by FU (27.5 mg/kg or 35 mg/kg). These treatments resulted in 3 severity levels of systemic toxicity indicated by changes in body weight. In addition to the well known FU+LV-induced diarrhea, myelosuppression, and stomatitis, facial edema, and enlargement of the submandibular salivary gland were consistently seen. Facial edema occurred almost exclusively in rats that subsequently underwent excessive weight loss and were euthanized. The submandibular, but not parotid or sublingual, salivary gland was enlarged and the severity of this effect changed in a bell-shaped relationship with respect to increasing FU+LV induced loss of body weight. Histologic examination of affected glands established the occurrence of bacterial infection, sialoadenitis and destruction of gland tissue. This paper provides the first known documentation of FU+LV treatment-induced selective pathology of the submandibular salivary gland. The selectivity of this toxicity, apparently not normally seen in humans, to the submandibular salivary gland of the rat is of interest and its mechanism warrants further investigation. PMID- 16036872 TI - Effects of medical therapy on pituitary tumors. AB - Previously surgery and irradiation were the only available procedures to treat patients with pituitary tumors. During the last few decades, novel drugs such as dopamine agonists and long-acting somatostatin analogs were developed and, an alternative medical therapy emerged. This paper summarizes the effect of medical therapy on the morphologic features of pituitary tumors and illustrates the ultrastructural alterations on electron micrographs. Currently drugs can be used in the management of pituitary tumors secreting GH, PRL, and/or TSH in excess. No medical therapy is available so far for ACTH-, FSH-, LH-, or alpha-subunit secreting tumors as well as non-hormone-secreting pituitary tumors. Dopamine agonists are effective in the management of PRL-secreting tumors; they cause marked reversible tumor shrinkage in the substantial majority of patients. Long acting somatostatin analogs are useful in the management of GH- and TSH-secreting pituitary tumors; they lead to mild to moderate tumor shrinkage in approximately 50% of cases. In patients treated with these drugs reduction of elevated blood hormone levels and amelioration of clinical symptoms ensue. It should be emphasized that no permanent cure is obtained. Blood hormone levels increase and the clinical symptoms reappear after discontinuation of treatment. Recently GH receptor blockers (pegvisomant) were introduced in the treatment of GH-producing pituitary adenomas. To the authors' knowledge the effect of these drugs on the morphology of pituitary tumors has not been revealed so far. PMID- 16036874 TI - The spectrum of pediatric tumors in infancy, childhood, and adolescence: a comprehensive review with emphasis on special techniques in diagnosis. AB - The spectrum of pediatric tumors varies considerably, from those derived from blastemal cells in various organ systems to proliferations of soft tissue supporting cells to hamartomatous processes that mimic malignant tumors. Small round cell tumors are often undifferentiated or poorly differentiated, making it difficult sometimes to provide a definitive diagnosis. Both benign and malignant tumors require a coordinated method for diagnosis, and need a comprehensive evaluation to provide the most appropriate diagnosis for designing therapy and predicting prognosis. Pediatric tumors require the integration of routine histopathologic examination with histochemical, immunocytochemical, ultrastructural, cytogenetic, and diagnostic molecular pathology techniques. This review provides updated guidelines with respect to the application of these special techniques in this rapidly evolving diagnostic arena. PMID- 16036873 TI - Morphological methods in the diagnosis of mitochondrial encephalomyopathies: the role of electron microscopy. AB - Mitochondrial encephalomyopathies (MEs) encompass a heterogeneous group of disorders that frequently present a diagnostic challenge to clinicians. Historically, MEs were diagnosed by finding ragged red fibers in the muscle biopsy and confirmatory evidence was provided by the presence of numerical and/or ultrastructural abnormalities in mitochondria. In most centers diagnosis involves clinical evaluation and the morphological, histochemical, and biochemical investigation of a skeletal muscle biopsy. However, with the availability of mitochondrial DNA analysis, the necessity and role of morphological methods and, in particular, electron microscopy has been questioned. The aim of this study was to delineate the role of electron microscopy in the diagnosis of MEs. PMID- 16036875 TI - Ultrastructural study of renal involvement in two females with Anderson-Fabry disease. AB - Anderson-Fabry disease (AFD) is a rare X-linked lipid storage disorder due to a deficient lysosomal a-galactosidase A (a-Gal) activity. In males with the classic form of the disease the enzymatic defect leads to progressive accumulation of glycosphingolipids (GL) in different organs, mainly in the kidney, heart, and brain, causing severe multisystem failure. AFD is usually mild in heterozygous females, but severe cerebrovascular, renal, and cardiac manifestations have been rarely described. The aim of this study is to describe renal involvement of mild symptomatic female carriers by ultrastructural analysis focusing to microvascular lesions, considered to be one of the major causes of systemic disease in AFD. Resin-embedded renal biopsies from 2 sisters with isolated mild proteinuria and belonging to a family group with AFD were observed by light and electron microscopy. In spite of the mild clinical symptoms, diffuse GL storages were demonstrated in all types of glomerular cells and in interstitial endothelial cells. Moreover, platelets were frequently observed in glomerular vassels, a feature coherent with a possible role of prothrombotic state, and platelet activation, in early glomerular lesions. PMID- 16036876 TI - Benign mullerian papilloma of childhood. AB - Benign mullerian papilloma of the female reproductive tract is a rare childhood tumor that can easily be mistaken by those unfamiliar with the entity for botryoid rhabdomyosarcoma. Ultrastructural findings have been mentioned only in two individual case reports, and these both were issued many years ago. The aim of this update is to familiarize the reader with the clinical, light, and electron microscopic features associated with this distinctive entity, and thereby hopefully preclude the risk of making a serious diagnostic error. Two cases are illustrated, one very typical in its presentation and the other less so. PMID- 16036877 TI - Primary ciliary dyskinesia: a review. AB - The entity sinusitis, bronchiectasis, and situs inversus is since long named Kartagener syndrome. Nowadays the designation used is primary ciliary dyskinesia (PCD), which implies cilia with decreased or total absence of motility, which may result in sinusitis, chronic bronchitis, bronchiectasis, and male infertility. A large number of deficiencies detectable on the ultrastructural level give rise to PCD. There may also be aberrations not detected up to the present. The normal left-right asymmetry of the body is thought to be due to the beating of the cilia in the embryonic (Hensen's) node. Total immotility of the cilia should therefore result in random asymmetry of the body that is situs inversus in 50% of the cases. It has also been claimed that 50% of cases with PCD have situs inversus. However, several deficiencies apparently do not cause total immotility, and all ultrastructural variants are not associated with situs inversus in 50% of the cases. Several of the deficiencies are difficult to detect. Optimal fixation and handling are therefore obligatory. The genetic changes behind the variants are now being studied in several laboratories. Patients with PCD have very low levels of nasal nitric oxide, which is of increasing diagnostic importance. Other established diagnostic methods are the saccharine test and determination of ciliary beat frequency. PMID- 16036878 TI - Cell death by autoschizis in TRAMP prostate carcinoma cells as a result of treatment by ascorbate: menadione combination. AB - A prostate carcinoma cell line derived from the transgenic murine prostate cancer model (TRAMP) was treated with ascorbate (VC) alone, menadione (VK(3)) alone, or a combination of ascorbate:menadione (VC + VK(3)) for 1, 2, and 4 h. Cytotoxic cell alterations examined by light and electron microscopy were treatment dependent with VC + VK(3) > VC > VK(3). Induced by oxidative stress, these alterations included cytokeletal changes conducive to cytoplasmic blebbing, self excisions, and progressive nuclear alterations. While the excised parts contained ribosomes, they were devoid of nuclear fragments or other organelles. The organelle-free self-excisions caused an extreme reduction in cell size as well as chromatolysis and karyolysis that were consistent with cell death by autoschizis, but not with apoptosis. PMID- 16036879 TI - Transmission electron microscopy in hematological diagnosis. AB - Transmission electron microscopy (TEM) is still a very useful adjunct for hematological diagnosis in the era of molecular techniques. In this article, the main applications of TEM to the cellular identification of normal myeloid cells, the study of dyserythropoietic conditions, myelodysplastic syndromes, congenital dyserythropoietic anemias, acute myeloid leukemias, and lymphoproliferative disorders, as well as the application of ultrastructural cytochemical reactions in hematological diagnosis, are reviewed. PMID- 16036881 TI - The evolving concept of renal neoplasia: impact of emerging molecular and electron microscopic studies. AB - The classification of renal tumors has evolved from one that initially encompassed only 2 types of tumors, i.e., clear and granular cell carcinomas, to the markedly expanded recent classification that incorporates new entities, some of which are primarily defined by specific molecular abnormalities. Despite these advances, a single tumor category, clear cell carcinoma, still incorporates the majority (approximately 70%) of renal tumors. It is, however, postulated that this single category is likely to encompass several different tumor types that are, at present, undifferentiated. Electron microscopic studies have been pivotal in defining the spectrum of oncocytoma-chromophobe renal cell carcinoma. Cytoplasmic eosinophilia found in some renal cell carcinomas currently classified as clear cell type is under intense study. Tumors that have recently emerged from this group include tumors with translocations involving chromosome Xp11.2, carcinomas associated with neuroblastoma and epithelioid angiomyolipoma. The spectrum of renal tumors seen in younger patients is wider than among older patients, with rare and unusual tumors being more likely seen in younger patients. The author concludes that although the routine application of electron microscopy to kidney tumor diagnosis may not be practical, systematic ultrastructural studies of these tumors may aid in the definition of new entities. PMID- 16036880 TI - Fatal pulmonary microsporidiosis due to encephalitozoon cuniculi following allogeneic bone marrow transplantation for acute myelogenous leukemia. AB - Microsporidia are ubiquitous obligate eukaryotic intracellular parasites that are now felt to be more akin to degenerate fungi than to protozoa. Microsporidia can be highly pathogenic, causing a broad range of symptoms in humans, especially individuals who are immunocompromised. The vast majority of human cases of microsporidiosis have been reported during the past 20 years, in patients with HIV/AIDS, while only relatively rare cases have been described in immunocompetent individuals. However, microsporidia infections are being increasingly reported in patients following solid-organ transplanation, where the main symptom has been diarrhea. The authors report the first case of pulmonary microsporidial infection in an allogeneic bone marrow transplant recipient in the United States and only the second case in the world. The patient, with a history of Hodgkin disease followed by acute myelogenous leukemia received a T-cell-depleted graft, but succumbed to respiratory failure 63 days post transplantation. An open lung biopsy, taken just before death, was originally thought to show toxoplasmosis. The correct diagnosis of microsporidiosis was made postmortem by light and electron microscopy. DNA polymerase chain reaction analysis confirmed the diagnosis and furthermore revealed it to be the dog strain of the microsporidia species Encephalitozoon cuniculi. Although to date rarely diagnosed, microsporidial infection should also be considered in the differential diagnosis of, e.g., unexplained pulmonary infection in bone marrow transplant patients. PMID- 16036882 TI - Mixed epithelial and stromal tumor of the kidney: preliminary immunohistochemical and electron microscopic studies of the epithelial component. AB - Mixed epithelial and stromal tumor of the kidney is a rare biphasic tumor composed of cysts and tubules embedded in the spindle cell stroma. Although the histogenesis of this tumor is unknown, it has been proposed that both components of the tumor, i.e., stromal and epithelial, are neoplastic. The authors report preliminary immunohistochemical and electron microscopic studies of the epithelial component from one case of a typical, benign, mixed epithelial, and stromal tumor of the kidney. In this study, some tubules showed positivity for proximal, while others showed positivity for distal, nephron immunomarkers. By electron microscopy, some tubules had features of proximal tubular epithelium, while other tubules had features of the loop of Henle (thin segments). The authors believe that in a benign tumor such morphologic heterogeneity is inconsistent with neoplastic proliferation. Therefore, they postulate that in mixed epithelial and stromal tumor of the kidney the tubules are entrapped rather than neoplastic. Additional studies are needed to address this issue and electron microscopy should play a significant role in this process. PMID- 16036883 TI - Merkel cells, normal and neoplastic: an update. AB - Merkel cells (MC) occur in the basal epidermal layer, hair follicles, and oral mucosa, as complexes with sensory axons. The axons transduce slowly adapting type I mechanoreception, and MC modulate their sensitivity. MC also determine and maintain the 3-dimensional epidermal structure. They have neuroendocrine granules, rigid spinous processes, and desmosomal junctions with each other and with keratinocytes. Rare MC are dermaWl. Current evidence supports a basal cell origin. Merkel cell carcinomas (MCC) occur mostly in sun-exposed skin in old age. Trabecular, intermediate, or small cell in pattern, MCC have neuroendocrine granules, intercellular junctions, rigid spinous processes, and a paranuclear collection of intermediate filaments staining for cytokeratin 20. Most MCC behave indolently, but those with the small cell pattern, and some with the intermediate pattern, are aggressive and rapidly fatal. PMID- 16036884 TI - The comparative role of immunohistochemistry and electron microscopy in the identification of myogenic differentiation in soft tissue pleomorphic sarcomas. AB - There is increasing evidence that histological classification of pleomorphic soft tissue sarcomas is prognostically useful, since a number of studies have provided evidence that myogenic differentiation is associated with a more aggressive clinical behavior. The aim of the current study was to analyze the role of electron microscopy in comparison with immunohistochemistry in the classification of soft tissue pleomorphic sarcomas. Thirty-nine pleomorphic sarcomas of the somatic soft tissues for which material for immunohistochemical and ultrastructural analysis was available were selected for this study. Cases were classified according to the criteria of the WHO classification of soft tissue tumors on the basis of the histologic appearance and of the results of immunohistochemical analysis, and then diagnoses were reconsidered at the light of the results of the ultrastructural analysis. The group of myogenic sarcomas included 13 leiomyosarcomas, 8 myofibrosarcomas, and 1 rhabdomyosarcoma, while the group of nonmyogenic sarcomas included 11 undifferentiated pleomorphic sarcomas/malignant fibrous histiocytomas (MFH), 4 myxofibrosarcomas, and 2 liposarcomas. Overall, there was a good concordance between immunohistochemistry and electron microscopy in recognizing myogenic differentiation in soft tissue pleomorphic sarcomas. Discrepancies included 1 case showing no immunoreactivity for muscle markers, which displayed ultrastructural features allowing reclassification as leiomyosarcoma, and 2 cases initially classified as undifferentiated pleomorphic sarcoma/MFH, which were reclassified as myofibrosarcomas after ultrastructural analysis. Ultrastructural analysis allowed the identification of pleomorphic sarcomas with myofibroblastic phenotype, a category that is not identifiable based on histologic and immunohistochemical profile. Notably, fibronexus junction was identified in tumor cells of 4 pleomorphic myofibrosarcomas, while 2 other lesions showed putative fibronexus junction structures, consisting of electron-dense straight fibrils adjacent to the cell surface, not clearly in continuity with cytoplasmic actin filaments. In conclusion, the results indicate that immunohistochemistry and electron microscopy can usefully complement each other in the classification of soft tissue pleomorphic sarcomas. PMID- 16036885 TI - Stromal elements for tumor diagnosis: a brief review of diagnostic electron microscopic features. AB - Tumor diagnosis mainly depends on the appearance of the tumor cells in recapitulating the appearance of primordial cells from which they arise. However, certain tumors may present with specific stromal changes that may assist/enhance the diagnosis. In this presentation, diagnostic stromal features have been reviewed. The cytoplasm is enclosed by a unit membrane, which serves as a barrier to, as well as an interface with, surrounding structures. Epithelial cells usually show characteristic basal-apical orientation. In mesenchymal tissue, different types of interface can be found in different types of mesenchymal tissue. External lamina can be defined as an anatomic structure, which encloses anatomic functional units. In epithelial tissue, cells in a functional unit are enclosed within a well-defined external lamina (EL). In malignant epithelial tumors, EL can become increasingly indistinct as tumors become less differentiated, and one has to look for it diligently. Within the external lamina, epithelial cells are closely packed with closely apposed cell membranes and cell attachment junctions. In contrast to epithelial tissue, mesenchymal tissue is usually characterized by the stromal elements they produce. Individual cells are embedded in the stroma, and individual mesenchymal cells represent the functional unit. Vascular endothelial cells are an exception since their relationship to stroma resembles to that of epithelial cells. Thus, tumors deriving from mesenchymal cells known to have external lamina such as muscle cells and Schwann cells tend to show total enclosure of cells by external lamina. In malignant muscle tumors, external lamina production can be focally present and found only by diligent search. In Schwann cell tumors, the presence of EL is prominent in low-grade tumors and more irregular and variable in malignant tumors. In the latter, stromal aggregation of scrolls of external lamina can be characteristic. Similar features are seen in ossifying fibromyxoid tumors. Fibronexus junctions (composed of extracellular fibronectin fillements linking intracellular 5-nm filaments) is claimed to be typical of myofbroblasts. Finding them in spindle cell tumors justifies a diagnosis of myofibroblastomas. There have been several stromal changes diagnostic for certain tumors found only by electron microscopy. Fibrous long-spaced collagen (known as Luse bodies) is diagnostic for peripheral nerve sheath tumors, but they can rarely be found in other tumors. Luse bodies usually appear as focally as crystallized aggregates apart from the regular collagenous interstitial stroma. They should be distinguished from other nonspecific long-spaced collagen changes. The changes are diffusely stromal in contrast to Luse bodies. Spiny collagen and amianthoid fibers are interesting collagen fibrils and their diagnostic value is questionable. Skeinoid fibers (SF) are short-spaced collagen of 41- to 45-nm banding so-named because of their peculiar appearance by electron microscopy simulating skeins of yarn. They were originally described in neurogenic tumors and small intestinal stromal tumors with features of gastrointestinal autonomic nerve tumors (GANT). Although there have been a few sporadic case reports of the presence of skeinoid fibers in nonneurogenic tumors, the frequent presence of SF in spindle cell tumors signifies their neurogenic nature in this authors' experience. An exception to this is that SF can be a constant element of rare ciliary body tumors known as ciliary mesectodermal leiomyomas, in which tumor cells show some resemblance to smooth muscle as well as Schwann cells. In addition to SF, several other types of peculiar crystallized collagen were observed in GANT tumors, particularly those with multiple tumor syndromes such as neurofibromatosis and Carney's triad. They simulate the appearance of railroad tracks or centrosomes. The reason for this is not known. The authors speculate that such collagen crystallization may be caused by genetic alterations involving collagenosis. Further studies will be necessary to clarify their pathogenesis. Another peculiar stromal change is electron-dense stromal filamentous aggregates with extra-long banding of > 250-nm periodicity previously described in Ewing sarcomas. This stromal change simulating a tiger skin pattern is also seen in primitive neuroectodermal tumors and malignant melanomas. In view of continually new discoveries of stromal changes that can be used for the differential diagnosis of tumors, the importance of close evaluation of stromal elements of tumors, and diligent application of electron microscopy in tumor diagnosis cannot be overemphasized. PMID- 16036886 TI - Applications of immunogold labeling in ultrastructural pathology. AB - This review provides an update on the use of postembedding immunogold labeling, preferentially of the protein A-gold technique, for electron microscopic research in diseased states. In the first part, some helpful antigen recovery techniques for use of immunogold labeling of ultrathin sections prepared from routinely aldehyde (and osmium tetroxide) fixed and conventionally epoxy resin-embedded tissue are cited. In the second part, selected applications for studies of tumor cell surface conjugates, such as polysialic acid in Wilms tumor, proinsulin insulin conversion in human insulinoma, and the importance of pre-Golgi intermediates for retention of a misfolded polypeptide hormone in a protein misfolding disease, are discussed. PMID- 16036887 TI - Two imaging techniques to find too flexible an airway: looking for malacia. PMID- 16036888 TI - Palivizumab for preterm infants. Is it worth it? AB - Respiratory syncytial virus infection is an important cause of morbidity. Although palivizumab prophylaxis is widely used, it is uncertain whether the cost is justified. A systematic review was therefore performed of the safety, efficacy, and the likely cost effectiveness of prophylaxis for preterm infants in the United Kingdom using a standard search strategy. The only randomised controlled trial identified showed a reduction in hospital admission but no benefit on more serious outcomes. None of the United Kingdom cost studies showed economic benefit for palivizumab prophylaxis. New treatments are rarely cost effective, and, in the absence of a comprehensive economic assessment, continued use for high risk infants may appear justified. PMID- 16036889 TI - The continuous glucose monitoring sensor in neonatal intensive care. AB - OBJECTIVE: To determine the feasibility of continuous glucose monitoring in the very low birthweight baby requiring intensive care, as these infants are known to be at high risk of abnormalities of glucose control. METHOD: Sixteen babies were studied from within 24 hours of delivery and for up to seven days. RESULTS: The subcutaneous glucose sensors were well tolerated and readings were comparable to those on near patient whole blood monitoring devices. CONCLUSION: Continuous glucose monitoring is practical in neonates, giving detailed information about glucose control. PMID- 16036890 TI - Prenatal three dimensional ultrasound detection of linear nevus sebaceous syndrome. PMID- 16036891 TI - Intrauterine programming of urinary calcium and magnesium excretion in children born to mothers with insulin dependent diabetes mellitus. AB - BACKGROUND: Offspring of diabetic rats have reduced urinary calcium and magnesium excretion compared with offspring of controls; these differences persist up to 16 weeks after birth, a time equivalent to young adulthood in humans. OBJECTIVES: To test the hypothesis that urinary calcium and magnesium excretion would be lower in children born to mothers with insulin dependent diabetes mellitus (ChMIDDM) than those born to non-diabetic mothers. METHODS: Concentrations of calcium, magnesium, sodium, and creatinine were measured in first void spot urine samples collected from 45 (28 male; median age 9.6 years) ChMIDDM and 127 (58 male; median age 11.3 years) controls. Analysis of covariance was used to test for differences in urinary calcium to creatinine ratios (UCa/Cr), magnesium to creatinine ratios (UMg/Cr), and log sodium to creatinine ratios (logUNa/Cr) between controls and ChMIDDM after allowing for the effects of sex and age. RESULTS: UCa/Cr (difference -0.10, 95% confidence interval (CI) -0.19 to -0.01; p = 0.03) and UMg/Cr (difference -0.15, 95% CI -0.22 to -0.08; p<0.0001) were lower in ChMIDDM than controls. However, logUNa/Cr did not differ between ChMIDDM and controls (difference -0.14, 95% CI -0.33 to 0.05; p = 0.1). The daily estimated intake of magnesium, sodium, and protein were significantly higher and that of calcium non-significantly higher in ChMIDDM than controls. In ChMIDDM, UCa/Cr and UMg/Cr were not related to diabetic control of mothers. CONCLUSIONS: Results of this study provide the first evidence that in humans, as in rats, there is modification of renal Ca and Mg handling in ChMIDDM, which persists well into childhood. PMID- 16036892 TI - Social deprivation and admission for neonatal care. AB - OBJECTIVE: To determine whether social deprivation is associated with neonatal unit admission. SETTING: English district general hospital. METHOD: Retrospective review of neonatal unit admission records between 1990 and 2002. RESULTS: There was a linear increase in admission rates with increasing deprivation. The admission rate was 6.1% of live births for infants in the most affluent quartile compared with 11.1% for those in the most deprived quartile. Admission rates for all indications except jaundice and feeding problems increased with increasing deprivation. CONCLUSION: Social deprivation correlates strongly with neonatal morbidity and the need for neonatal unit admission. This finding has implications for professionals in public health and primary and secondary care. PMID- 16036893 TI - Does antenatal pelvic dilation predict renal scarring? AB - Moderate antenatal renal pelvic dilation (5-15 mm) may suggest vesicoureteric reflux, but it is not known to predict renal scarring. Dimercaptosuccinic acid scans on such children aged over 4 years showed a scarring rate (0/133 boys, 1/56 girls) similar to our local population. Investigation and treatment of moderate dilation may not be required. PMID- 16036894 TI - Quantitative ultrasound assessment of bone in preterm and term neonates. AB - There is a need to explore novel methods of assessing bone health in sick preterm infants. This study of the speed of sound in the long bones of newborn term and preterm infants shows that, in this population, this technique is not site specific and has a high degree of interobserver and intraobserver precision. The speed of sound in newborn infants is primarily dependent on gestation rather than birth weight. PMID- 16036895 TI - Nasal continuous positive airway pressure: does bubbling improve gas exchange? AB - In a randomised crossover trial, 26 babies, treated with Hudson prong continuous positive airway pressure (CPAP) from a bubbling bottle, received vigorous, high amplitude, or slow bubbling for 30 minutes. Pulse oximetry, transcutaneous carbon dioxide, and respiratory rate were recorded. The bubbling rates had no effect on carbon dioxide, oxygenation, or respiratory rate. PMID- 16036896 TI - Ignac Semmelweis (1818-1865) of Budapest and the prevention of puerperal fever. AB - As a junior obstetrician in Vienna in 1847, Semmelweis discovered the cause of puerperal fever and introduced a method for its prevention. Although his findings were hailed by some as comparable to Jenner's introduction of vaccination, for various reasons his ideas failed to gain general acceptance until after his death. PMID- 16036897 TI - Funisitis and raised interleukin 6 concentrations in gastric aspirates at birth. PMID- 16036901 TI - A perspective on the muscle reflex: implications for congestive heart failure. AB - In this review we examine the exercise pressor reflex in health and disease. The role of metabolic and mechanical stimulation of thin fiber muscle afferents is discussed. The role ATP and lactic acid play in stimulating and sensitizing these afferents is examined. The role played by purinergic receptors subdivision 2, subtype X, vanilloid receptor subtype 1, and acid-sensing ion channels in mediating the effects of ATP and H+ are discussed. Muscle reflex activation in heart failure is then examined. Data supporting the concept that the metaboreflex is attenuated and that the mechanoreflex is accentuated are presented. The role the muscle mechanoreflex plays in evoking renal vasoconstriction is also described. PMID- 16036902 TI - Energy cost and muscular activity required for leg swing during walking. AB - To investigate the metabolic cost and muscular actions required for the initiation and propagation of leg swing, we applied a novel combination of external forces to subjects walking on a treadmill. We applied a forward pulling force at each foot to assist leg swing, a constant forward pulling force at the waist to provide center of mass propulsion, and a combination of these foot and waist forces to evaluate leg swing. When the metabolic cost and muscle actions were at a minimum, the condition was considered optimal. We reasoned that the difference in energy consumption between the optimal combined waist and foot force trial and the optimal waist force-only trial would reflect the metabolic cost of initiating and propagating leg swing during normal walking. We also reasoned that a lower muscle activity with these assisting forces would indicate which muscles are normally responsible for initiating and propagating leg swing. With a propulsive force at the waist of 10% body weight (BW), the net metabolic cost of walking decreased to 58% of normal walking. With the optimal combination, a propulsive force at the waist of 10% BW plus a pulling force at the feet of 3% BW the net metabolic cost of walking further decreased to 48% of normal walking. With the same combination, the muscle activity of the iliopsoas and rectus femoris muscles during the swing phase was 27 and 60% lower, respectively, but the activity of the medial gastrocnemius and soleus before swing did not change. Thus our data indicate that approximately 10% of the net metabolic cost of walking is required to initiate and propagate leg swing. Additionally, the hip flexor muscles contribute to the initiation and propagation leg swing. PMID- 16036903 TI - Chronic hypoxia modulates relations among calcium, myosin light chain phosphorylation, and force differently in fetal and adult ovine basilar arteries. AB - The present study tests the hypothesis that age-related differences in contractility of cerebral arteries from hypoxic animals involve changes in myofilament Ca2+ sensitivity. Basilar arteries from term fetal and nonpregnant adult sheep maintained 110 days at 3,820 m were used in measurements of cytosolic calcium concentration ([Ca2+]i), myosin light chain phosphorylation, and contractile tensions induced by graded concentrations of K+ or serotonin (5-HT). Slopes relating [Ca2+]i to tension were similar in fetal (0.83 +/- 0.07) and adult (1.02 +/- 0.08) arteries for K+ contractions but were significantly greater for fetal (3.77 +/- 0.64) than adult (2.00 +/- 0.13) arteries for 5-HT contractions. For both K+ and 5-HT contractions, these relations were left shifted in fetal compared with adult arteries, indicating greater Ca2+ sensitivity in fetal arteries. In contrast, slopes relating [Ca2+]i and %myosin phosphorylation for K+ contractions were less in fetal (0.37 +/- 0.08) than adult (0.81 +/- 0.07) arteries, and the fetal curves were right shifted. For 5-HT contractions, the slope of the Ca2+-phosphorylation relation was similar in fetal (0.33 +/- 0.09) and adult (0.33 +/- 0.23) arteries, indicating that 5-HT depressed Ca2+-induced myosin phosphorylation in adult arteries. For slopes relating %myosin phosphorylation and tension, fetal values (K+: 1.52 +/- 0.22, 5 HT: 7.66 +/- 1.70) were less than adult values (K+: 2.13 +/- 0.30, 5-HT: 8.29 +/- 2.40) for both K+- and 5-HT-induced contractions, although again fetal curves were left shifted relative to the adult. Thus, in hypoxia-acclimatized basilar arteries, a downregulated ability of Ca2+ to promote myosin phosphorylation is offset by an upregulated ability of phosphorylated myosin to produce force yielding an increased fetal myofilament Ca2+ sensitivity. Postnatal maturation reprioritizes the mechanisms regulating hypoxic contractility through changes in the source of activator Ca2+, the pathways governing myosin light chain phosphorylation, and its interaction with actin. PMID- 16036905 TI - Expressional reprogramming of survival pathways in rat cardiocytes by neuregulin 1beta. AB - Neuregulin/ErbB2-induced kinase signaling provides essential survival and protection clues for functional integrity of the adult heart and skeletal muscle. To define the regulatory pathways involved in neuregulin-dependent muscle cell survival, we set out to map the largely unknown transcript targets of this growth/differentiation factor in cardiocytes. Freshly isolated adult primary rat cardiocytes were treated for 24 h with recombinant human neuregulin-1beta (NRG 1beta, 30 ng/ml). Transcript level alterations in NRG-1beta-treated and control cardiocytes (n = 6) were identified with Atlas Rat Toxicology 1.2 cDNA arrays (BD Clontech) and established permutation L1 regression analysis. Selected transcriptional adjustments were confirmed by RT-PCR and Western blotting. Involvement of MAPK pathways was verified with the inhibitor PD-98059. Application of the single dose of NRG-1beta to quiescent cardiocytes induced expressional reprogramming of distinct cellular processes. This response included a prominent 50-100% increase in transcripts of multiple redox systems. It also involved a comparable mRNA augmentation of protein synthetic and folding factors together with augmented message for the trigger of cardiac hypertrophy, cyclin D1 (CCND1). First evidence for a role of neuregulin in promotion of mitochondrial turnover, voltage-gated ion channel expression, and the suppression of fatty acid transporter mRNAs was revealed. Subsequent analysis confirmed a corresponding upregulation of redox factor proteins thioredoxin and the thioredoxin reductase 1, GSTP-1, and CCND1 and demonstrated downregulation of the related transcripts by PD-98059 in neuregulin-stimulated cultures. These MAPK-dependent expressional adjustments point to novel oxidative defense and hypertrophy pathways being involved in the longer lasting protective function of neuregulin in the heart. PMID- 16036904 TI - Firing rates of motor units in human vastus lateralis muscle during fatiguing isometric contractions. AB - We investigated the firing rate of motor units in the vastus lateralis muscle in five healthy young men (mean = 21.4 yr, SD = 0.9) during a sequence of isometric constant-torque contractions repeated to exhaustion. The contractions were sustained at 20% of the maximal voluntary level, measured at the beginning of the test sequence. Electromyographic (EMG) signals were recorded via quadrifilar fine wire electrodes and subsequently decomposed into their constituent motor unit action potentials to obtain the motor unit firing times. In addition, we measured the whole muscle mechanical properties during the fatigue task using electrical stimulation. The firing rate of motor units first decreased within the first 10 20% of the endurance time of the contractions and then increased. The firing rate increase was accompanied by recruitment of additional motor units as the force output remained constant. The elicited twitch and tetanic torque responses first increased and then decreased. The two processes modulated in a complementary fashion at the same time. Our data suggest that, when the vastus lateralis muscle is activated to maintain a constant torque output, its motoneuron pool receives a net excitatory drive that first decreases to compensate for the short-lived potentiation of the muscle force twitch and then increases to compensate for the diminution of the force twitch. The underlying inverse relationship between the firing rate and the recruitment threshold that has been reported for nonfatigued contractions is maintained. We, therefore, conclude that the central nervous system control of vastus lateralis motor units remains invariant during fatigue in submaximal isometric isotonic contractions. PMID- 16036906 TI - Contraction signaling to glucose transport in skeletal muscle. AB - Contracting skeletal muscles acutely increases glucose transport in both healthy individuals and in people with Type 2 diabetes, and regular physical exercise is a cornerstone in the treatment of the disease. Glucose transport in skeletal muscle is dependent on the translocation of GLUT4 glucose transporters to the cell surface. It has long been believed that there are two major signaling mechanisms leading to GLUT4 translocation. One mechanism is insulin-activated signaling through insulin receptor substrate-1 and phosphatidylinositol 3-kinase. The other is an insulin-independent signaling mechanism that is activated by contractions, but the mediators of this signal are still unknown. Accumulating evidence suggests that the energy-sensing enzyme AMP-activated protein kinase plays an important role in contraction-stimulated glucose transport. However, more recent studies in transgenic and knockout animals show that AMP-activated protein kinase is not the sole mediator of the signal to GLUT4 translocation and suggest that there may be redundant signaling pathways leading to contraction stimulated glucose transport. The search for other possible signal intermediates is ongoing, and calcium, nitric oxide, bradykinin, and the Akt substrate AS160 have been suggested as possible candidates. Further research is needed because full elucidation of an insulin-independent signal leading to glucose transport would be a promising pharmacological target for the treatment of Type 2 diabetes. PMID- 16036907 TI - Exercise-induced increase in muscle insulin sensitivity. AB - Exercise/muscle contraction activates glucose transport. The increase in muscle glucose transport induced by exercise is independent of insulin. As the acute effect of exercise on glucose transport wears off, it is replaced by an increase in insulin sensitivity. An increase in insulin sensitivity results in a shift in the insulin dose-response curve to the left, with a decrease in the concentration of insulin needed to induce 50% of the maximal response. This phenomenon, which plays a major role in rapid muscle glycogen accumulation after exercise, is not mediated by amplification of the insulin signal. Development of the increase in insulin sensitivity after contractions does not require protein synthesis or activation of p38 MAPK. It does require the presence of a serum protein during the period of contractile activity. The effect of exercise on muscle insulin sensitivity is mimicked by hypoxia and by treatment of muscles with 5 aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside to activate AMP-activated protein kinase. The postexercise increase in sensitivity of muscle glucose transport to activation is not specific for insulin but also involves an increased susceptibility to activation by a submaximal contraction/hypoxia stimulus. The increase in insulin sensitivity is mediated by translocation of more GLUT4 glucose transporters to the cell surface in response to a submaximal insulin stimulus. Although the postexercise increase in muscle insulin sensitivity has been characterized in considerable detail, the basic mechanisms underlying this phenomenon remain a mystery. PMID- 16036908 TI - Point: The muscle pump raises muscle blood flow during locomotion. PMID- 16036909 TI - BOLD indirect vs. ASL direct measurement of muscle perfusion. PMID- 16036910 TI - A different analysis applied to a mathematical model on output of exhaled nitric oxide. PMID- 16036911 TI - Chemical rescue of I-site cleavage in living cells and in vitro discriminates between the cytomegalovirus protease, assemblin, and its precursor, pUL80a. AB - Chemical rescue is an established approach that offers a directed strategy for designing mutant enzymes in which activity can be restored by supplying an appropriate exogenous compound. This method has been used successfully to study a broad range of enzymes in vitro, but its application to living systems has received less attention. We have investigated the feasibility of using chemical rescue to make a conditional-lethal mutant of the cytomegalovirus (CMV) maturational protease. The 28-kDa CMV serine protease, assemblin, has a Ser-His His catalytic triad and an internal (I) cleavage site near its midpoint. We found that imidazole can restore I-site cleavage to mutants inactivated by replacing the critical active site His with Ala or with Gly, which rescued better. Comparable rescue was observed for counterpart mutants of the human and simian CMV assemblin homologs and occurred in both living cells and in vitro. Cleavage was established to be at the correct site by amino acid sequencing and proceeded at approximately 11%/h in bacteria and approximately 30%/h in vitro. The same mutations were unresponsive to chemical rescue in the context of the assemblin precursor, pUL80a. This catalytic difference distinguishes the two forms of the CMV protease. PMID- 16036912 TI - Discovery, structural determination, and putative processing of the precursor protein that produces the cyclic trypsin inhibitor sunflower trypsin inhibitor 1. AB - Backbone-cyclized proteins are becoming increasingly well known, although the mechanism by which they are processed from linear precursors is poorly understood. In this report the sequence and structure of the linear precursor of a cyclic trypsin inhibitor, sunflower trypsin inhibitor 1 (SFTI-1) from sunflower seeds, is described. The structure indicates that the major elements of the reactive site loop of SFTI-1 are present before processing. This may have importance for a protease-mediated cyclizing reaction as the rigidity of SFTI-1 may drive the equilibrium of the reaction catalyzed by proteolytic enzymes toward the formation of a peptide bond rather than the normal cleavage reaction. The occurrence of residues in the SFTI-1 precursor susceptible to cleavage by asparaginyl proteases strengthens theories that involve this enzyme in the processing of SFTI-1 and further implicates it in the processing of another family of plant cyclic proteins, the cyclotides. The precursor reported here also indicates that despite strong active site sequence homology, SFTI-1 has no other similarities with the Bowman-Birk trypsin inhibitors, presenting interesting evolutionary questions. PMID- 16036913 TI - Synthetic lethal interaction of the mitochondrial phosphatidylethanolamine and cardiolipin biosynthetic pathways in Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae mitochondria contain enzymes required for synthesis of the phospholipids cardiolipin (CL) and phosphatidylethanolamine (PE), which are enriched in mitochondrial membranes. Previous studies indicated that PE may compensate for the lack of CL, and vice versa. These data suggest that PE and CL have overlapping functions and that the absence of both lipids may be lethal. To address this hypothesis, we determined whether the crd1delta mutant, which lacks CL, was viable in genetic backgrounds in which PE synthesis was genetically blocked. Deletion of the mitochondrial PE pathway gene PSD1 was synthetically lethal with the crd1delta mutant, whereas deletion of the Golgi and endoplasmic reticulum pathway genes PSD2 and DPL1 did not result in synthetic lethality. A 20 fold reduction in phosphatidylcholine did not affect the growth of crd1delta cells. Supplementation with ethanolamine, which led to increased PE synthesis, or with propanolamine, which led to synthesis of the novel phospholipid phosphatidylpropanolamine, failed to rescue the synthetic lethality of the crd1delta psd1delta cells. These results suggest that mitochondrial biosynthesis of PE is essential for the viability of yeast mutants lacking CL. PMID- 16036914 TI - Origin of informational polymers. Differential stability of 3'- and 5' phosphoester bonds in deoxy monomers and oligomers. AB - To survive, an informational macromolecule must solve the major problem set by its very polymeric nature: instability. This is especially true in prebiotic terms because of the presumed initial absence of protective structures (proteins, lipids, etc.). We have analyzed the stability of the beta-glycosidic and of the 3'- and 5'-phosphoester bonds in both deoxy monomers and deoxy oligomers under a large set of conditions. The results show a strong dependence of the relative stability of these bonds on the physico-chemical environment. A set of conditions has been identified in which the stability of polymers becomes comparable with that of the precursor monomers. In certain instances the stability of the 5' phosphoester bond is even higher in the polymer than in the mononucleotide. PMID- 16036915 TI - Dominant negative mechanism underlies autosomal dominant Stargardt-like macular dystrophy linked to mutations in ELOVL4. AB - ELOVL4 (elongation of very long chain fatty acids 4) is a member of the ELO family of proteins involved in the biosynthesis of very long chain fatty acids. Protein truncation mutations in ELOVL4 have been identified in patients with autosomal dominant Stargardt-like macular degeneration. To determine whether a dominant negative mechanism is responsible for the autosomal dominant inheritance pattern of this disease, we studied the subcellular localization and interaction of wild type and mutant ELOVL4 in COS-7 and HEK 293T cultured cells by immunofluorescence and co-immunoprecipitation. Wild type ELOVL4 containing an endoplasmic reticulum retention sequence was localized to the endoplasmic reticulum as expected. In contrast, disease-associated C-terminal truncation ELOVL4 mutants accumulated as large inclusions exhibiting aggresome-like characteristics in a juxtanuclear position within COS-7 cells. When the wild type and mutant proteins were co-expressed incultured cells, wild type ELOVL4 co purified with mutant ELOVL4 on an immunoaffinity column and co-localized with the mutant protein in aggresome-like inclusions adjacent to the nucleus. These results indicate that wild type and mutant ELOVL4 form a complex that exhibits an abnormal subcellular localization found for individually expressed mutant ELOVL4. From these studies, we conclude that disease-linked C-terminal truncation mutants of ELOVL4 exert a dominant negative effect on wild type ELOVL4, altering its subcellular localization. This dominant negative mechanism contributes to the autosomal dominant inheritance of Stargardt-like macular dystrophy. PMID- 16036916 TI - Overexpression of NBS1 contributes to transformation through the activation of phosphatidylinositol 3-kinase/Akt. AB - Nijmegen breakage syndrome (NBS) is a chromosomal instability syndrome associated with cancer predisposition, radiosensitivity, microcephaly, and growth retardation. The NBS gene product, NBS1 (p95) or nibrin, is a part of the hMre11 complex, a central player associated with double strand break repair. We previously demonstrated that c-Myc directly activates NBS1 expression. Here we have shown that constitutive expression of NBS1 in Rat1a and HeLa cells induces/enhances their transformation. Repression of endogenous NBS1 levels using short interference RNA reduces the transformation activity of two tumor cell lines. Increased NBS1 expression is observed in 40-52% of non-small cell lung carcinoma, hepatoma, and esophageal cancer samples. NBS1 overexpression stimulates phosphatidylinositol (PI) 3-kinase activity, leading to increased phosphorylation levels of Akt and its downstream targets such as glycogen synthase kinase 3beta and mammalian target of rapamycin in different cell lines and tumor samples. Transformation induced by NBS1 overexpression can be inhibited by a PI3-kinase inhibitor (LY294002). Repression of endogenous Akt expression by short interference RNA decreases the transformation activity of Rat1a cells overexpressing NBS1. These results indicate that overexpression of NBS1 is an oncogenic event that contributes to transformation through the activation of PI3 kinase/Akt. PMID- 16036918 TI - Mechanistic insights into the isochorismate pyruvate lyase activity of the catalytically promiscuous PchB from combinatorial mutagenesis and selection. AB - PchB from Pseudomonas aeruginosa possesses isochorismate pyruvate lyase (IPL) and weak chorismate mutase (CM) activity. Homology modeling based on a structurally characterized CM, coupled with randomization of presumed key active site residues (Arg54, Glu90, Gln91) and in vivo selection for CM activity, was used to derive mechanistic insights into the IPL activity of PchB. Mutation of Arg54 was incompatible with viability, and the CM and IPL activities of an engineered R54K variant were reduced 1,000-fold each. The observation that position 90 was tolerant to substitution but position 91 was essentially confined to Gln or Glu in functional variants rules out involvement of Glu90 in general base catalysis. Counter to the generally accepted mechanistic hypothesis for pyruvate lyases, we propose for PchB a rare [1,5]-sigmatropic reaction mechanism that invokes electrostatic catalysis in analogy to the [3,3]-pericyclic rearrangement of chorismate in CMs. A common catalytic principle for both PchB functions is also supported by the covariance of the catalytic parameters for the CM and IPL activities and the shared functional requirement for a protonated Glu91 in Q91E variants. The experiments demonstrate that focusing directed evolution strategies on the readily accessible surrogate activity of an enzyme can provide valuable insights into the mechanism of the primary reaction. PMID- 16036917 TI - Identical phenotypes of CatSper1 and CatSper2 null sperm. AB - Among several candidate Ca(2+) entry channels in sperm, only CatSper1 and CatSper2 are known to have required roles in male fertility. Past work with CatSper1 null sperm indicates that a critical lesion in hyperactivated motility underlies the infertility phenotype and is associated with an absence of depolarization-evoked Ca(2+)entry. Here we show that failure of hyperactivation of CatSper2 null sperm similarly correlates with an absence of depolarization evoked Ca(2+) entry. Additional shared aspects of the phenotypes of CatSper1 and 2 null sperm include unperturbed regional distributions of conventional voltage gated Ca(2+) channel proteins and robust acceleration of the flagellar beat by bicarbonate. Further study reveals that treatment of both wild-type and CatSper2 null sperm with procaine increases beat asymmetry, a characteristic of the flagellar waveform of hyperactivation. This partial rescue of the loss-of hyperactivation phenotype suggests that an absence of CatSper2 precludes hyperactivation by preventing delivery of needed Ca(2+) messenger rather than by preventing flagellar responses to Ca(2+). CatSper2 null sperm also have an increased basal cAMP content and beat frequency. Protein kinase A inhibitor H89 lowers beat frequency to that of wild-type sperm, suggesting that CatSper2 is required for protein kinase A-mediated, tonic control of resting cAMP content. Relative to wild-type testis, CatSper1 and -2 null testes contain normal amounts of CatSper2 and -1 transcripts, respectively. However, CatSper1 null sperm lack CatSper2 protein and CatSper2 null sperm lack CatSper1 protein. Hence, stable expression of CatSper1 protein requires CatSper2 and vice versa. This co dependent expression dictates identical loss-of-function sperm phenotypes for CatSper1 and -2 null mutants. PMID- 16036919 TI - Phosphoinositide 3-kinase signaling to Akt promotes keratinocyte differentiation versus death. AB - Signaling pathways regulating the differentiation program of epidermal cells overlap widely with those activated during apoptosis. How differentiating cells remain protected from premature death, however, is still poorly defined. We show here that the phosphoinositide 3-kinase (PI3K)/Akt pathway is activated at early stages of mouse keratinocyte differentiation both in culture and in the intact epidermis in vivo. Expression of active Akt in keratinocytes promotes growth arrest and differentiation, whereas pharmacological blockade of PI3K inhibits the expression of "late" differentiation markers and leads to death of cells that would otherwise differentiate. Mechanistically, the activation of the PI3K/Akt pathway in keratinocyte differentiation depends on the activity of the epidermal growth factor receptor and Src families of tyrosine kinases and the engagement of E-cadherin-mediated adhesion. During this process, PI3K associates increasingly with cadherin-catenin protein complexes bearing tyrosine phosphorylated YXXM motifs. Thus, the PI3K signaling pathway regulates the choice between epidermal cell differentiation and death at the cross-talk between tyrosine kinases and cadherin-associated catenins. PMID- 16036920 TI - Ligand-assisted aggregation of proteins. Dimerization of serum amyloid P component by bivalent ligands. AB - A comprehensive series of solution and crystallographic studies reveal how simple, achiral, bivalent ligands of the cyclic pyruvate of glycerol promote face to-face complex formation of the pentraxin, serum amyloid P component (SAP) into decamers. SAP, a protein of the human innate immune system, is universally present in amyloids, including cerebral amyloid deposits found in the brain of Alzheimer disease patients. Removal of SAP through a specific aggregation mechanism mediated by multivalent ligands appears to provide therapeutic benefit in the progression of this disease. Crystallographic studies reveal that in our novel series of ligands only the methyl and carboxylate moieties of the pyruvate ketal directly interact with the protein, but the geometric constraints imposed by the tether dictate which of two chair conformations are adopted by the pyruvate dioxane ring. Solution studies, as interpreted through a simple thermodynamic model, account for the distribution of pentameric and decameric bound states at different ligand concentrations and indicate that differences in the flexibility of the tether determine the geometry and stability of the specific aggregates formed between SAP and two different bivalent ligands. The factors affecting the design of ligands promoting face-to-face protein dimerization as well as potential biological implications are discussed. PMID- 16037064 TI - CCM1 and CCM2 protein interactions in cell signaling: implications for cerebral cavernous malformations pathogenesis. AB - Cerebral cavernous malformations (CCMs) are sporadically acquired or inherited vascular lesions of the central nervous system consisting of clusters of dilated thin-walled blood vessels that predispose individuals to seizures and stroke. Familial CCM is caused by mutations in KRIT1 (CCM1) or in malcavernin (CCM2), the murine ortholog of which was concurrently characterized as osmosensing scaffold for MEKK3 (OSM). The roles of the CCM proteins in the pathogenesis of the disorder remain largely unknown. Here, we use co-immunoprecipitation, fluorescence resonance energy transfer and subcellular localization strategies to show that the CCM1 gene product, KRIT1, interacts with the CCM2 gene product, malcavernin/OSM. Analogous to the established interactions of CCM1 and beta1 integrin with ICAP1, the CCM1/CCM2 association is dependent upon the phosphotyrosine binding (PTB) domain of CCM2. A familial CCM2 missense mutation abrogates the CCM1/CCM2 interaction, suggesting that loss of this interaction may be critical in CCM pathogenesis. CCM2 and ICAP1 bound to CCM1 via their respective PTB domains differentially influence the subcellular localization of CCM1. Furthermore, we expand upon the established involvement of CCM2 in the p38 mitogen-activated protein kinase signaling module by demonstrating that CCM1 associates with CCM2 and MEKK3 in a ternary complex. These data indicate that the genetic heterogeneity observed in familial CCM may reflect mutation of different molecular members of a coordinated signaling complex. PMID- 16037065 TI - Analysis of intronic conserved elements indicates that functional complexity might represent a major source of negative selection on non-coding sequences. AB - The non-coding portion of human genome is punctuated by a large number of multispecies conserved sequence (MCS) elements with largely unknown function. We demonstrate that MCSs are unevenly distributed in human introns with the majority of relatively short introns (< 9 kb long) displaying no or a few MCSs and that MCS density reaching up to 10% of total size in longer introns. After correction for intron length, MCSs were found to be enriched within genes involved in development and transcription, whereas depleted in immune response loci. Moreover, many central nervous system tissues show a preferential expression of MCS-rich genes and MCS enrichment significantly correlates with gene functional complexity in terms of distinct protein domains. Analysis of human-mouse orthologous pairs indicated a significant association between intronic MCS density and conservation of protein sequence, promoter regions and untranslated sequences. Moreover, MCS density correlates with the predicted occurrence of human-mouse conserved alternative splicing events. These observations suggest that evolution acts on human genes as integrated units of coding and regulatory capacity and that functional complexity might represent a major source of negative selection on non-coding sequences. To substantiate our result, we also searched previously experimentally identified intronic regulatory elements and indicate that about half of these sequences map to an MCS; in particular, support to the notion whereby mutations in MCSs can result in human genetic diseases is provided, because three previously identified intronic pathological variations were found to occur within MCSs, and human disease and cancer genes were found significantly enriched in MCSs. PMID- 16037066 TI - Neuron-specific relaxation of Igf2r imprinting is associated with neuron-specific histone modifications and lack of its antisense transcript Air. AB - The mouse insulin-like growth factor II receptor (Igf2r) gene and its antisense transcript Air are reciprocally imprinted in most tissues, but in the brain, Igf2r is biallelically expressed despite the imprinted Air expression. To investigate the molecular mechanisms of such brain-specific relaxation of Igf2r imprinting, we analyzed its expression and epigenetic modifications in neurons, glial cells and fibroblasts by the use of primary cortical cell cultures. In glial cells and fibroblasts, Igf2r was maternally expressed and Air was paternally expressed, whereas in the primary cultured neurons, Igf2r was biallelically expressed and Air was not expressed. In the differentially methylated region 2 (DMR2), which includes the Air promoter, allele-specific DNA methylation, differential H3 and H4 acetylation and H3K4 and K9 di-methylation were maintained in each cultured cell type. In DMR1, which includes the Igf2r promoter, maternal-allele-specific DNA hypomethylation, histones H3 and H4 acetylation and H3K4 di-methylation were apparent in glial cells and fibroblasts. However, in neurons, biallelic DNA hypomethylation and biallelic histones H3 and H4 acetylation and H3K4 di-methylation were detected. These data indicate that lack of reciprocal imprinting of Igf2r and Air in the brain results from neuron specific relaxation of Igf2r imprinting associated with neuron-specific histone modifications in DMR1 and lack of Air expression. Our observation of biallelic Igf2r expression with no Air expression in neurons sheds light on the function of Air as a critical effector in Igf2r silencing and suggests that neuron-specific epigenetic modifications related to the lineage determination of neural stem cells play a critical role in controlling imprinting by antisense transcripts. PMID- 16037067 TI - A dinucleotide deletion in the ankyrin promoter alters gene expression, transcription initiation and TFIID complex formation in hereditary spherocytosis. AB - Ankyrin defects are the most common cause of hereditary spherocytosis (HS). In some HS patients, mutations in the ankyrin promoter have been hypothesized to lead to decreased ankyrin mRNA synthesis. The ankyrin erythroid promoter is a member of the most common class of mammalian promoters which lack conserved TATA, initiator or other promoter cis elements and have high G+C content, functional Sp1 binding sites and multiple transcription initiation sites. We identified a novel ankyrin gene promoter mutation, a TG deletion adjacent to a transcription initiation site, in a patient with ankyrin-linked HS and analyzed its effects on ankyrin expression. In vitro, the mutant promoter directed decreased levels of gene expression, altered transcription initiation site utilization and exhibited defective binding of TATA-binding protein (TBP) and TFIID complex formation. In a transgenic mouse model, the mutant ankyrin promoter led to abnormalities in gene expression, including decreased expression of a reporter gene and altered transcription initiation site utilization. These data indicate that the mutation alters ankyrin gene transcription and contributes to the HS phenotype by decreasing ankyrin gene synthesis via disruption of TFIID complex interactions with the ankyrin core promoter. These studies support the model that in promoters that lack conserved cis elements, the TFIID complex directs preinitiation complex formation at specific sites in core promoter DNA and provide the first evidence that disruption of TBP binding and TFIID complex formation in this type of promoter leads to alterations in start site utilization, decreased gene expression and a disease phenotype in vivo. PMID- 16037068 TI - Guidelines for conducting and reporting whole genome/large-scale association studies. PMID- 16037069 TI - Guidelines for association studies in Human Molecular Genetics. PMID- 16037071 TI - IL-2 responsiveness of CD4 and CD8 lymphocytes: further investigations with human IL-2Rbeta transgenic mice. AB - Responsiveness to IL-2 varies from one lympho-mononuclear subset to another. NK lymphocytes and monocytes spontaneously respond to IL-2 whereas it is generally accepted that T and B lymphocytes need to be activated to fully acquire this competence. To further investigate this phenomenon, we studied human IL-2Rbeta (hIL-2Rbeta) transgenic mice constitutively expressing heterospecific, intermediate-affinity IL-2R (hIL-2Rbeta/mouse IL-2Rgamma(c)). We noted that the B lymphocytes and monocytes from spleens of these hIL-2Rbeta transgenic animals failed to grow when cultured in IL-2-containing medium. Under the same experimental conditions, CD4 lymphocytes survived, again without growth, whereas CD8 lymphocytes and NK cells were able to proliferate and develop potent LAK cytotoxicity. The properties of these CD4 and CD8 lymphocytes were then compared after purification. Both subsets expressed functional IL-2R able to induce global protein phosphorylation and, more precisely, signal transducer and activation of transcription 5 and Erk phosphorylation. Therefore, the differential growth potential of these CD4 and CD8 lymphocytes cannot be explained by the lack of IL 2R-dependent early signaling events. When the entrance of purified CD4 and CD8 lymphocytes into the cell cycle was analyzed, we found that the CD4 lymphocytes were unable to enter the G1 phase in the absence of anti-CD3 stimulation. This correlates with the effect of IL-2 on cyclin-dependent kinase inhibitor p27(kip1). In CD4 lymphocytes, IL-2 does not affect p27(kip1) expression. But in CD8 lymphocytes, IL-2 down-modulates p27(kip1). These results indicate that, aside from IL-2R expression and function, IL-2 responsiveness is also controlled by lineage-specific mechanisms. PMID- 16037072 TI - Characterization of mouse spleen cells by subtractive proteomics. AB - Major analytical challenges encountered by shotgun proteome analysis include both the diversity and dynamic range of protein expression. Often new instrumentation can provide breakthroughs in areas where other analytical improvements have not been successful. In the current study, we utilized new instrumentation (LTQ FT) to characterize complex protein samples by shotgun proteomics. Proteomic analyses were performed on murine spleen tissue separated by magnetic beads into distinct CD45- and CD45+ cell populations. Using shotgun protein analysis we identified approximately 2,000 proteins per cell group by over 12,000 peptides with mass deviations of less than 4.5 ppm. Datasets obtained by LTQ FT analysis provided a significant increase in the number of proteins identified and greater confidence in those identifications and improved reproducibility in replicate analyses. Because CD45- and not CD45+ cells are able to regenerate functional pancreatic islet cells in a mouse model of type I diabetes, protein expression was further compared by a subtractive proteomic approach in search of an exclusive protein expression profile in CD45- cells. Characterization of the proteins exclusively identified in CD45- cells was performed using gene ontology terms via the Javascript GoMiner. The CD45- cell subset readily revealed proteins involved in development, suggesting the persistence of a fetal stem cell in an adult animal. PMID- 16037073 TI - Protein complexes in the archaeon Methanothermobacter thermautotrophicus analyzed by blue native/SDS-PAGE and mass spectrometry. AB - Methanothermobacter thermautotrophicus is a thermophilic archaeon that produces methane as the end product of its primary metabolism. The biochemistry of methane formation has been extensively studied and is catalyzed by individual enzymes and proteins that are organized in protein complexes. Although much is known of the protein complexes involved in methanogenesis, only limited information is available on the associations of proteins involved in other cell processes of M. thermautotrophicus. To visualize and identify interacting and individual proteins of M. thermautotrophicus on a proteome-wide scale, protein preparations were separated using blue native electrophoresis followed by SDS-PAGE. A total of 361 proteins, corresponding to almost 20% of the predicted proteome, was identified using peptide mass fingerprinting after MALDI-TOF MS. All previously characterized complexes involved in energy generation could be visualized. Furthermore the expression and association of the heterodisulfide reductase and methylviologen-reducing hydrogenase complexes depended on culture conditions. Also homomeric supercomplexes of the ATP synthase stalk subcomplex and the N5 methyl-5,6,7,8-tetrahydromethanopterin:coenzyme M methyltransferase complex were separated. Chemical cross-linking experiments confirmed that the multimerization of both complexes was not experimentally induced. A considerable number of previously uncharacterized protein complexes were reproducibly visualized. These included an exosome-like complex consisting of four exosome core subunits, which associated with a tRNA-intron endonuclease, thereby expanding the constituency of archaeal exosomes. The results presented show the presence of novel complexes and demonstrate the added value of including blue native gel electrophoresis followed by SDS-PAGE in discovering protein complexes that are involved in catabolic, anabolic, and general cell processes. PMID- 16037074 TI - An assessment of hepatitis C virus infection among health-care workers of the National Cancer Institute of Naples, Southern Italy. AB - BACKGROUND: As many people with chronic hepatitis C virus (HCV) infection are asymptomatic, HCV infection could spread easily among the health-care workers of the National Cancer Institute of Naples (especially before the identification of HCV and in the absence of good, effective preventative measures, e.g. sterile syringe use, gloves, protective glasses). METHODS: In order to determine whether there is a transmission risk for HCV infection from patient to health-care worker, we carried out a cross-sectional study of a cohort of National Cancer Institute health-care workers in Naples, Southern Italy. RESULTS: The chi2-test was not significant; we did not find any significant risk for HCV in the 'other untrained staff' group [odds ratio (OR) 2.2; 95% confidence interval (CI) 0.4 10.9] or in the health-care workers group (OR 1.6; 95% CI 0.4-7.0). In the health care worker subgroups, doctors were the reference category because of the low prevalence of HCV in this subgroup (3.3%). A non-significant association was found in the professional nurses group (OR 2.7; 95% CI 0.8-8.8), as well as in the categories of technicians and biologists. CONCLUSIONS: No excessive risk was found among the health-care workers as a whole or among the different categories of personnel, confirming that health-care employees have benefited sufficiently from preventative measures. PMID- 16037075 TI - Recreational drug consumers: who seeks treatment? AB - BACKGROUND: Recreational drug consumption represents a complex issue, because of the lack of a shared definition, settings and patterns of consumptions, and poorness of evidence-based treatments. In spite of the great number of users, just few seek specific treatment, probably representing those at major risk of continuing and problematic use. The general objective of this study was to report the characteristics and main consumption patterns of recreational drug consumers treated in Regione Piemonte (Italy), in order to formulate hypotheses suitable for further research aimed at establishing more effective preventive intervention and treatment. METHODS: All addiction treatment services in the region were required to fill in, for each user, a questionnaire reporting: contact of the user with the service, socio-demographic data, drugs consumption and interventions/treatments. RESULTS: Among 441 users, 70.3% were new users, 85.5% were males and the average age of consumption onset was 22 years. Psychiatric and traumatics events were present in 13.3% and in 17.9% of cases, respectively. The most frequent primary substances were cocaine (76.9%) and ecstasy (13.8%). Substances taken in association with primary drug are quoted in 75% of cases. CONCLUSIONS: The users that get in touch with services are those showing greater problems, representing, therefore, the main target for preventive public health interventions. Some interesting points can be noted: among patients treated, cocaine represents the main issue; females seem to be more sensitive to recreational drug effects; social class (education), settings and use habits are similar for recreational drugs users and heroin users. PMID- 16037076 TI - Gender and regional differences in perceived job stress across Europe. AB - BACKGROUND: Over the last 20 years stress at work has been found to be predictive of several conditions such as coronary heart disease, high blood pressure and non specific sick leave. The Karasek demand/control/strain concept has been the most widely used in prospective epidemiological studies. OBJECTIVES: To describe distribution in Karasek's demand/control (DC) dimensions as well as prevalence of strain in samples from different parts of Europe grouped into three regions (South, Middle, Sweden), adjusting for occupation. To describe gender differences in Karasek's DC dimensions along with strain prevalence and assess the regional stability of those differences in different occupational groups. DESIGN: The Job stress, Absenteeism and Coronary heart disease in Europe (JACE) study, a Concerted Action (Biomed I) of the European Union, is a multicentre prospective cohort epidemiological study: 38,019 subjects at work aged 35-59 years were surveyed at baseline. Standardised techniques were used for occupation coding (International Standardised Classification of Occupations) and for the DC model (Karasek scale): five items for the psychological demand and nine items for the control or decision latitude dimensions, respectively. RESULTS: A total of 34,972 subjects had a complete data set. There were important regional differences in the Karasek scales and in prevalence of strain even after adjustment for occupational class. Mean demand and control were higher in the Swedish centres when compared to two centres in Milano and Barcelona (Southern region) and values observed in four centres (Ghent, Brussels, Lille and Hoofddorp) in Middle Europe were closer to those observed in the Southern cities than to those obtained in the Swedish cities. Clerks (ISCO 4) and, more specifically, office clerks (ISCO 41) exhibited the smallest regional variation. In a multivariate model, the factor 'region' explained a small fraction of total variance. In the two Southern centres as well as in the four Middle European centres, men perceived marginally less job-demand as compared to women whereas the reverse was observed in the two Swedish centres. Differences were larger for control: men appeared to perceive more control at work than did women. In a multivariate model, gender explained a small fraction whereas occupational level explained a large fraction of the variance. CONCLUSIONS: In this standardised multicentre European study Karasek's DC model showed large gender and occupational differences whereas geographic region explained a small fraction of the total DC variance, notwithstanding large differences in labour market and working conditions as pointed out by the European Commission as recently as 2000. PMID- 16037077 TI - Mortality among 723,948 foreign- and native-born Swedes 1970-1999. AB - BACKGROUND: Mortality in a population is regarded as an accurate and valid measure of the population's health. There are a few international studies, predominantly cross-sectional, of mortality among all foreign-born compared with an indigenous population, and the results have varied. No Swedish longitudinal study describing and analysing mortality data was found in a literature review. METHODS: This study describes and analyses the differences in mortality between foreign-born persons and native Swedes during the period 1970-1999, based on data from Statistics Sweden and the National Board of Health and Welfare. The database consisted of 723,948 persons, 361 974 foreign-born living in Sweden in 1970, aged > or = 16 years, and 361 974 Swedish controls matched for age, sex, occupation and type of employment, living in the same county in 1970. RESULTS: The results showed increased mortality for foreign-born persons compared with the Swedish controls [odds ratio (OR) 1.08; 95% confidence interval (CI) 1.07-1.08]. Persons who had migrated 'late' (1941-1970) to Sweden were 2.5 years younger at time of death than controls. In relation to country of birth, the highest risk odds were for men born in Finland (OR 1.21), Denmark (OR 1.11) and Norway/Iceland (OR 1.074). Age cohorts of foreign-born persons born between 1901 and 1920 had higher mortality at age 55-69 years than cohorts born between 1921 and 1944. CONCLUSIONS: Migrants had higher mortality than the native population, and migration may be a risk factor for health; therefore, this seems to be an important factor to consider when studying mortality and health. PMID- 16037078 TI - The impact of the European Environment and Health Process on UK environment and health policy, plans and practice: what difference has it made? AB - The European Environment and Health Process (EEHP), led by the World Health Organization (WHO) Regional Office for Europe, aims to support WHO Member States as they plan and implement national and international environment and health policies. An evaluation of the impact of the EEHP in the UK was conducted in preparation for the fourth Ministerial Conference on Environment and Health in Budapest, 2004. The evaluation identified a number of impacts and influences of the EEHP. This concluded that the process had only a marginal direct influence on policy within the UK. However, it was also concluded that the process had resulted in several indirect influences, including better cooperation between government departments, greater awareness of environment and health issues from an international perspective, and a higher political profile of environment and health issues. A few outcomes of the EEHP also appear to have been taken into account in some national and local policy documents. The National Environmental Health Action Plan, which was produced as a direct result of the EEHP, appears to have had little direct impact in the UK, probably because of the lack of an implementation process and indicators, and because it was superseded by other policy initiatives relatively soon after publication. A need for better coordination and promotion of the EEHP amongst stakeholders responsible for environment and health policy areas was also identified. PMID- 16037079 TI - Relationship between economic development and risk of injuries in older adults and the elderly. A global analysis of unintentional injury mortality in an epidemiologic transition perspective. AB - BACKGROUND: Injury is recognized as an important public health problem not only in high-income countries, but also in low- and middle-income countries. Other studies have shown inverse association between economic development and unintentional injury mortality (UIM) among children and young adults. In this study we aimed to examine the association between economic development and UIM rates among 45+ years-old populations within the theoretical framework of epidemiologic transition. METHODS: The World Health Statistics Annual 1996 was used as the main source of data. The relationship between UIM rates and gross national product (GNP) per capita was studied by using two methods: (i) with regression analysis, and (ii) by categorizing the data in four income-based country groups and the comparison of differences in their mean values. The ages were divided into four sub-groups: 45-54 years, 55-64 years, 65-74 years and 75+ years. RESULTS: UIM rates were inversely correlated with GNP per capita in 45-54, 55-64 and 65-74 year-olds for both sexes. On the other hand, among the 75+ years populations for both sexes, the association between GNP per capita and UIM rates was positive (male, r = 0.234, and female, r = 0.337). CONCLUSION: Our study suggests that UIM rates shows deviating patterns across age groups. UIM exhibits declining rates among younger age groups by economic development which is consistent with what could be expected from the epidemiologic transition model, while injuries in older groups appear to increase by economic development. These findings imply that unintentional injury, as a whole, is not a homogeneous phenomenon from an epidemiological transition perspective. PMID- 16037080 TI - Non-synaptic mechanisms underlie the after-effects of cathodal transcutaneous direct current stimulation of the human brain. AB - Although cathodal transcranial direct current stimulation (tDCS) decreases cortical excitability, the mechanisms underlying DC-induced changes remain largely unclear. In this study we investigated the effect of cathodal DC stimulation on spontaneous neural activity and on motor responses evoked by stimulation of the central and peripheral nervous system. We studied 17 healthy volunteers. Transcranial magnetic stimulation (TMS) and transcranial electrical stimulation (TES) of the motor area were used to study the effects of cathodal tDCS (1.5 mA, 10 min) on resting motor threshold and motor evoked potentials (MEPs) recorded from the contralateral first dorsal interosseous muscle (FDI). The electroencephalographic (EEG) activity in response to cathodal tDCS was analysed by power spectral density (PSD). Motor axonal excitability changes in response to transcutaneous DC stimulation of the ulnar nerve (0.3 mA, 10 min) were assessed by testing changes in the size of the compound muscle action potential (CMAP) elicited by submaximal nerve stimulation. Cathodal tDCS over the motor area for 10 min increased the motor threshold and decreased the size of MEPs evoked by TMS for at least 60 min after current offset (t(0) 71.7 +/- 5%, t(20) 50.8 +/- 11%, t(40) 47.7 +/- 7.7%, and t(60) 39.7 +/- 6.4%, P < 0.01). The tDCS also significantly decreased the size of MEPs elicited by TES (t(0) 64 +/- 16.4%, P = 0.09; t(20) 67.6 +/- 10.8%, P = 0.06; and t(40) 58.3 +/- 9.9%, P < 0.05). At the same time in the EEG the power of delta (2-4 Hz) and theta (4-7 Hz) rhythms increased (delta 181.1 +/- 40.2, P < 0.05; and theta 138.7 +/- 27.6, P = 0.07). At the peripheral level cathodal DC stimulation increased the size of the ulnar nerve CMAP (175 +/- 34.3%, P < 0.05). Our findings demonstrate that the after-effects of tDCS have a non-synaptic mechanism of action based upon changes in neural membrane function. These changes apart from reflecting local changes in ionic concentrations, could arise from alterations in transmembrane proteins and from electrolysis-related changes in [H(+)] induced by exposure to constant electric field. PMID- 16037081 TI - Activation of transient receptor potential vanilloid 1 (TRPV1) by resiniferatoxin. AB - Transient receptor potential vanilloid 1 (TRPV1) is a Ca(2+) permeable non selective cation channel activated by physical and chemical stimuli. Resiniferatoxin (RTX), an ultrapotent agonist of TRPV1, is under investigation for treatment of urinary bladder hyper-reflexia and chronic pain conditions. Here, we have determined the characteristics of RTX-induced responses in cells expressing native and cloned rat TRPV1. Whole-cell currents increase with repeated application of submaximal concentrations of RTX until a maximal response is attained and do not deactivate even after prolonged washout. Interestingly, the rate of activation and block by capsazepine of RTX-induced currents are significantly slower than for capsaicin-induced currents. RTX-induced whole-cell currents are outwardly rectifying, but to a lesser extent than capsaicin-induced currents. RTX-induced single channel currents exhibit multiple conductance states and outward rectification. The open probability (P(o)) of RTX-induced currents is higher at all potentials as compared to capsaicin-induced currents, which showed a strong voltage-dependent decrease at negative potentials. Single-channel kinetic analyses reveal that open-time distribution of RTX-induced currents can be fitted with three exponential components at negative and positive potentials. The areas of distribution of the longer open time constants are significantly larger than capsaicin-induced currents. The closed-time distribution of RTX induced currents can be fitted with three exponential components as compared to capsaicin-induced currents, which require four exponential components. Current clamp experiments reveal that low concentrations of RTX caused a slow and sustained depolarization beyond threshold while generating few action potentials. Concentrations of capsaicin required for the same extent of depolarization generated a significantly greater number of action potentials. These properties of RTX may play a role in its clinical usefulness. PMID- 16037082 TI - Influence of ionic strength on the time course of force development and phosphate release by dogfish muscle fibres. AB - We measured the effects of ionic strength (IS), 200 (standard) and 400 mmol l(-1) (high), on force and ATP hydrolysis during isometric contractions of permeabilized white fibres from dogfish myotomal muscle at their physiological temperature, 12 degrees C. One goal was to test the validity of our kinetic scheme that accounts for energy release, work production and ATP hydrolysis. Fibres were activated by flash photolysis of the P(3)-1-(2 nitrophenyl) ethyl ester of ATP (NPE-caged ATP), and time-resolved phosphate (P(i)) release was detected with the fluorescent protein MDCC-PBP, N-(2[1-maleimidyl]ethyl)-7 diethylamino-coumarin-3-carboxamide phosphate binding protein. High IS slowed the transition from rest to contraction, but as the fibres approached the isometric force plateau they showed little IS sensitivity. By 0.5 s of contraction, the force and the rate of P(i) release at standard and high IS values were not significantly different. A five-step reaction mechanism was used to account for the observed time courses of force and P(i) release in all conditions explored here. Only the rate constants for reactions of ATP, ADP and P(i) with the contractile proteins varied with IS, thus suggesting that the actin-myosin interactions are largely non-ionic. Our reaction scheme also fits previous results for intact fibres. PMID- 16037083 TI - Fetal brain hypometabolism during prolonged hypoxaemia in the llama. AB - In this study we looked for additional evidence to support the hypothesis that fetal llama reacts to hypoxaemia with adaptive brain hypometabolism. We determined fetal llama brain temperature, Na(+) and K(+) channel density and Na(+)-K(+)-ATPase activity. Additionally, we looked to see whether there were signs of cell death in the brain cortex of llama fetuses submitted to prolonged hypoxaemia. Ten fetal llamas were instrumented under general anaesthesia to measure pH, arterial blood gases, mean arterial pressure, heart rate, and brain and core temperatures. Measurements were made 1 h before and every hour during 24 h of hypoxaemia (n = 5), which was imposed by reducing maternal inspired oxygen fraction to reach a fetal arterial partial pressure of oxygen (P(a,O(2))) of about 12 mmHg. A normoxaemic group was the control (n = 5). After 24 h of hypoxaemia, we determined brain cortex Na(+)-K(+)-ATPase activity, ouabain binding, and the expression of NaV1.1, NaV1.2, NaV1.3, NaV1.6, TREK1, TRAAK and K(ATP) channels. The lack of brain cortex damage was assessed as poly ADP-ribose polymerase (PARP) proteolysis. We found a mean decrease of 0.56 degrees C in brain cortex temperature during prolonged hypoxaemia, which was accompanied by a 51% decrease in brain cortex Na(+)-K(+)-ATPase activity, and by a 44% decrease in protein content of NaV1.1, a voltage-gated Na(+) channel. These changes occurred in absence of changes in PARP protein degradation, suggesting that the cell death of the brain was not enhanced in the fetal llama during hypoxaemia. Taken together, these results provide further evidence to support the hypothesis that the fetal llama responds to prolonged hypoxaemia with adaptive brain hypometabolism, partly mediated by decreases in Na(+)-K(+)-ATPase activity and expression of NaV channels. PMID- 16037084 TI - Aminoglycoside ototoxicity: permeant drugs cause permanent hair cell loss. PMID- 16037085 TI - Retrograde endocannabinoid regulation of GABAergic inhibition in the rat dentate gyrus granule cell. AB - The dentate gyrus is a key input gateway for the hippocampus, and dentate function is potently regulated by GABAergic inhibition. GABAergic inhibition is plastic and modulated by many factors. Cytoplasmic calcium ([Ca(+)](i)) is one of these factors, and its elevation inhibits GABA-mediated transmission in the hippocampus including the dentate gyrus granule cells (DGCs). We examined whether the [Ca(+)](i)-dependent decrease of GABA(A) receptor-mediated inhibitory postsynaptic current (IPSC) is explained by the retrograde suppression of GABA release caused by the depolarization-induced elevation of [Ca(+)](i) in DGCs (DSI: depolarization-induced suppression of inhibition). Repeated brief depolarizations or a single long depolarization inhibited spontaneous IPSCs with amplitudes over 25 pA for up to a minute, and reduced the amplitude of IPSCs evoked by direct stimulation in the molecular layer, suggesting that DGCs are susceptible to DSI. The magnitude of DSI correlated linearly with the duration of depolarization, and so did the increase of [Ca(+)](i). DSI was blocked by intrapipette application of BAPTA. In addition, bath application of thapsigargin and ryanodine, and intrapipette application of ryanodine and ruthenium red reduced the [Ca(+)](i) increase caused by the DSI-inducing depolarization, and substantially reduced the magnitude of DSI. Finally, the cannabinoid receptor agonists, CP55,942 and WIN55,212-2, mimicked DSI and prevented further IPSC reduction by DSI. DSI was blocked by the antagonist, SR141716A. We conclude that GABAergic inhibition in DGCs is subject to endogenous cannabinoid (eCB)-mediated retrograde regulation, and this process involves a depolarization-initiated release of Ca(+) from ryanodine-sensitive stores. Our findings suggest eCBs probably have physiological functions in the regulation of GABAergic plasticity in the dentate gyrus. PMID- 16037086 TI - Functional interconnectivity between the globus pallidus and the subthalamic nucleus in the mouse brain slice. AB - In accordance with its central role in basal ganglia circuitry, changes in the rate of action potential firing and pattern of activity in the globus pallidus (GP)-subthalamic nucleus (STN) network are apparent in movement disorders. In this study we have developed a mouse brain slice preparation that maintains the functional connectivity between the GP and STN in order to assess its role in shaping and modulating bursting activity promoted by pharmacological manipulations. Fibre-tract tracing studies indicated that a parasagittal slice cut 20 deg to the midline best preserved connectivity between the GP and the STN. IPSCs and EPSCs elicited by electrical stimulation confirmed connectivity from GP to STN in 44/59 slices and from STN to GP in 22/33 slices, respectively. In control slices, 74/76 (97%) of STN cells fired tonically at a rate of 10.3 +/- 1.3 Hz. This rate and pattern of single spiking activity was unaffected by bath application of the GABA(A) antagonist picrotoxin (50 microM, n = 9) or the glutamate receptor antagonist (6-cyano-7-nitroquinoxaline-2, 3-dione (CNQX) 10 microM, n = 8). Bursting activity in STN neurones could be induced pharmacologically by application of NMDA alone (20 microm, 3/18 cells, 17%) but was more robust if NMDA was applied in conjunction with apamin (20-100 nM, 34/77 cells, 44%). Once again, neither picrotoxin (50 microM, n = 5) nor CNQX (10 microM, n = 5) had any effect on the frequency or pattern of the STN neurone activity while paired STN and GP recordings of tonic and bursting activity show no evidence of coherent activity. Thus, in a mouse brain slice preparation where functional GP-STN connectivity is preserved, no regenerative synaptically mediated activity indicative of a dynamic network is evident, either in the resting state or when neuronal bursting in both the GP and STN is generated by application of NMDA/apamin. This difference from the brain in Parkinson's disease may be attributed either to insufficient preservation of cortico-striato-pallidal or cortico-subthalamic circuitry, and/or an essential requirement for adaptive changes resulting from dopamine depletion for the expression of network activity within this tissue complex. PMID- 16037087 TI - Functional role of Na+-HCO3- cotransport in migration of transformed renal epithelial cells. AB - Cell migration is crucial for immune defence, wound healing or formation of tumour metastases. It has been shown that the activity of the Na(+)-H(+) exchanger (NHE1) plays an important role in cell migration. However, so far it is unknown whether Na(+)- HCO(3)(-) cotransport (NBC), which has similar functions in the regulation of intracellular pH (pH(i)) as NHE1, is also involved in cell migration. We therefore isolated NHE-deficient Madin-Darby canine kidney (MDCK-F) cells and tested whether NBC compensates for NHE in pH(i) and cell volume regulation as well as in migration. Intracellular pH was measured with the fluorescent pH indicator 2'7'-bis(carboxyethyl)-5-carboxyfluorescein (BCECF). The expression of NBC isoforms was determined with semiquantitative PCR. Migration was monitored with time-lapse video microscopy and quantified as the displacement of the cell centre. We found that MDCK-F cells express the isoform NBC1 (SLCA4A gene product) at a much higher level than the isoform kNBC3 (SLCA4A8 gene product). This difference is even more pronounced in NHE-deficient cells so that NBC1 is likely to be the major acid extruder in these cells and the major mediator of propionate-induced cell volume increase. NHE-deficient MDCK-F cells migrate more slowly than normal MDCK-F cells. NBC activity promotes migration during an acute intracellular acid load and increases migratory speed and displacement on a short timescale (< 30 min) whereas it has no effect on the long term behaviour of migrating MDCK-F cells. Taken together, our results show that NBC actvity, despite many functional similarities, does not have the same importance for cell migration as NHE1 activity. PMID- 16037088 TI - Autocrine activation of nicotinic acetylcholine receptors contributes to Ca2+ spikes in mouse myotubes during myogenesis. AB - It is widely accepted that nicotinic acetylcholine receptor (nAChR) channel activity controls myoblast fusion into myotubes during myogenesis. In this study we explored the possible role of nAChR channels after cell fusion in a murine cell model. Using videoimaging techniques we showed that embryonic muscle nAChR channel openings contribute to the spontaneous transients of intracellular concentration of Ca2+ ([Ca2+]i) and to twitches characteristic of developing myotubes before innervation. Moreover, we observed a choline acetyltransferase immunoreactivity in the myotubes and we detected an acetylcholine-like compound in the extracellular solution. Therefore, we suggest that the autocrine activation of nAChR channels gives rise to [Ca2+]i spikes and contractions. Spontaneous openings of the nAChR channels may be an alternative, although less efficient, mechanism. We report also that blocking the nAChRs causes a significant reduction in cell survival, detectable as a decreased number of myotubes in culture. This led us to hypothesize a possible functional role for the autocrine activation of the nAChRs. By triggering mechanical activity, such activation could represent a strategy to ensure the trophism of myotubes in the absence of nerves. PMID- 16037089 TI - High intensity exercise decreases global brain glucose uptake in humans. AB - Physiological activation increases glucose uptake locally in the brain. However, it is not known how high intensity exercise affects regional and global brain glucose uptake. The effect of exercise intensity and exercise capacity on brain glucose uptake was directly measured using positron emission tomography (PET) and [18F]fluoro-deoxy-glucose ([18F]FDG). Fourteen healthy, right-handed men were studied after 35 min of bicycle exercise at exercise intensities corresponding to 30, 55 and 75% of on three separate days. [18F]FDG was injected 10 min after the start of the exercise. Thereafter exercise was continued for another 25 min. PET scanning of the brain was conducted after completion of the exercise. Regional glucose metabolic rate (rGMR) decreased in all measured cortical regions as exercise intensity increased. The mean decrease between the highest and lowest exercise intensity was 32% globally in the brain (38.6+/-4.6 versus 26.1+/-5.0 micromol (100 g)-1 min-1, P<0.001). Lactate availability during exercise tended to correlate negatively with the observed brain glucose uptake. In addition, the decrease in glucose uptake in the dorsal part of the anterior cingulate cortex (37% versus 20%, P<0.05 between 30% and 75% of VO2max) was significantly more pronounced in subjects with higher exercise capacity. These results demonstrate that brain glucose uptake decreases with increase in exercise intensity. Therefore substrates other than glucose, most likely lactate, are utilized by the brain in order to compensate the increased energy needed to maintain neuronal activity during high intensity exercise. Moreover, it seems that exercise training could be related to adaptive metabolic changes locally in the frontal cortical regions. PMID- 16037090 TI - Connexin-specific cell-to-cell transfer of short interfering RNA by gap junctions. AB - The purpose of this study was to determine whether oligonucleotides the size of siRNA are permeable to gap junctions and whether a specific siRNA for DNA polymerase beta (pol beta) can move from one cell to another via gap junctions, thus allowing one cell to inhibit gene expression in another cell directly. To test this hypothesis, fluorescently labelled oligonucleotides (morpholinos) 12, 16 and 24 nucleotides in length were synthesized and introduced into one cell of a pair using a patch pipette. These probes moved from cell to cell through gap junctions composed of connexin 43 (Cx43). Moreover, the rate of transfer declined with increasing length of the oligonucleotide. To test whether siRNA for pol beta was permeable to gap junctions we used three cell lines: (1) NRK cells that endogenously express Cx43; (2) Mbeta16tsA cells, which express Cx32 and Cx26 but not Cx43; and (3) connexin-deficient N2A cells. NRK and Mbeta16tsA cells were each divided into two groups, one of which was stably transfected to express a small hairpin RNA (shRNA), which gives rise to siRNA that targets pol beta. These two pol beta knockdown cell lines (NRK-kcdc and Mbeta16tsA-kcdc) were co-cultured with labelled wild type, NRK-wt or Mbeta16tsA-wt cells or N2A cells. The levels of pol beta mRNA and protein were determined by semiquantitative RT-PCR and immunoblotting. Co-culture of Mbeta16tsA-kcdc cells with Mbeta16tsA-wt, N2A or NRK-wt cells had no effect on pol beta levels in these cells. Similarly, co culture of NRK-kcdc with N2A cells had no effect on pol beta levels in the N2A cells. In contrast, co-culture of NRK-kcdc with NRK-wt cells resulted in a significant reduction in pol beta in the wt cells. The inability of Mbeta16tsA kcdc cells to transfer siRNA is consistent with the fact that oligonucleotides of the 12 nucleotide length were not permeable to Cx32/Cx26 channels. This suggested that Cx43 but not Cx32/Cx26 channels allowed the cell-to-cell movement of the siRNA. These results support the novel hypothesis that non-hybridized and possible hybridized forms of siRNA can move between mammalian cells through connexin-specific gap junctions. PMID- 16037091 TI - Light responses and light adaptation in rat retinal rods at different temperatures. AB - Rod responses to brief pulses of light were recorded as electroretinogram (ERG) mass potentials across isolated, aspartate-superfused rat retinas at different temperatures and intensities of steady background light. The objective was to clarify to what extent differences in sensitivity, response kinetics and light adaptation between mammalian and amphibian rods can be explained by temperature and outer-segment size without assuming functional differences in the phototransduction molecules. Corresponding information for amphibian rods from the literature was supplemented by new recordings from toad retina. All light intensities were expressed as photoisomerizations per rod (Rh*). In the rat retina, an estimated 34% of incident photons at the wavelength of peak sensitivity caused isomerizations in rods, as the (hexagonally packed) outer segments measured 1.7 microm x 22 microm and had specific absorbance of 0.016 microm(-1) on average. Fractional sensitivity (S) in darkness increased with cooling in a similar manner in rat and toad rods, but the rat function as a whole was displaced to a ca 0.7 log unit higher sensitivity level. This difference can be fully explained by the smaller dimensions of rat rod outer segments, since the same rate of phosphodiesterase (PDE) activation by activated rhodopsin will produce a faster drop in cGMP concentration, hence a larger response in rat than in toad. In the range 15-25 degrees C, the waveform and absolute time scale of dark-adapted dim-flash photoresponses at any given temperature were similar in rat and toad, although the overall temperature dependence of the time to peak (t(p)) was somewhat steeper in rat (Q(10) approximately 4 versus 2-3). Light adaptation was similar in rat and amphibian rods when measured at the same temperature. The mean background intensity that depressed S by 1 log unit at 12 degrees C was in the range 20-50 Rh* s(-1) in both, compared with ca 4500 Rh* s( 1) in rat rods at 36 degrees C. We conclude that it is not necessary to assume major differences in the functional properties of the phototransduction molecules to account for the differences in response properties of mammalian and amphibian rods. PMID- 16037092 TI - Balance between cardiac output and sympathetic nerve activity in resting humans: role in arterial pressure regulation. AB - Large, reproducible interindividual differences exist in resting sympathetic nerve activity among normotensive humans with similar arterial pressures, resulting in a lack of correlation between muscle sympathetic nerve activity (MSNA) and arterial pressure among individuals. Although it is known that the arterial pressure is the main short-term determinant of MSNA in humans via the arterial baroreflex, the lack of correlation among individuals suggests that the level of arterial pressure is not the only important input in regulation of MSNA in humans. We studied the relationship between cardiac output (CO) and baroreflex control of sympathetic activity by measuring MSNA (peroneal microneurography), arterial pressure (arterial catheter), CO (acetylene uptake technique) and heart rate (HR; electrocardiogram) in 17 healthy young men during 20 min of supine rest. Across individuals, MSNA did not correlate with mean or diastolic blood pressure (r<0.01 for both), but displayed a significant negative correlation with CO (r=-0.71, P=0.001). To assess whether CO is related to arterial baroreflex control of MSNA, we constructed a baroreflex threshold diagram for each individual by plotting the percentage occurrence of a sympathetic burst against diastolic pressure. The mid-point of the diagram (T50) at which 50% of cardiac cycles are associated with bursts, was inversely related to CO (r=-0.75, P<0.001) and stroke volume (SV) (r=-0.57, P=0.015). We conclude that dynamic inputs from CO and SV are important in regulation of baroreflex control of MSNA in healthy, normotensive humans. This results in a balance between CO and sympathetically mediated vasoconstriction that may contribute importantly to normal regulation of arterial pressure in humans. PMID- 16037093 TI - Presynaptic N-type and P/Q-type Ca2+ channels mediating synaptic transmission at the calyx of Held of mice. AB - At the nerve terminal, both N- and P/Q-type Ca2+ channels mediate synaptic transmission, with their relative contribution varying between synapses and with postnatal age. To clarify functional significance of different presynaptic Ca2+ channel subtypes, we recorded N-type and P/Q-type Ca2+ currents directly from calyces of Held nerve terminals in alpha1A-subunit-deficient mice and wild-type (WT) mice, respectively. The most prominent feature of P/Q-type Ca2+ currents was activity-dependent facilitation, which was absent for N-type Ca2+ currents. EPSCs mediated by P/Q-type Ca2+ currents showed less depression during high-frequency stimulation compared with those mediated by N-type Ca2+ currents. In addition, the maximal inhibition by the GABAB receptor agonist baclofen was greater for EPSCs mediated by N-type channels than for those mediated by P/Q-type channels. These results suggest that the developmental switch of presynaptic Ca2+ channels from N- to P/Q-type may serve to increase synaptic efficacy at high frequencies of activity, securing high-fidelity synaptic transmission. PMID- 16037094 TI - Electrophysiological characterization of native Na+-HCO3- cotransporter current in bovine parotid acinar cells. AB - Using patch-clamp and molecular biological techniques, we identified and characterized membrane currents most likely generated by an electrogenic Na+-HCO3 cotransporter (NBCe) in acutely dissociated bovine parotid acinar (BPA) cells. When BPA cells were dialysed with a N-methyl-D-glucamine (NMDG)-glutamate-rich pipette solution, switching a Na-glutamate-rich, nominally HCO3--free bath solution to the one containing 25 mM HCO3-, but not Cl-, elicited a whole-cell current with a linear current-voltage relation. The HCO3- evoked current was abolished by total replacement of extracellular Na+ (Na+o) with NMDG or by 0.5 mM 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid (DIDS), and was only partially supported by Li+o, but not by K+o, Cs+o, and cholineo. The reversal potential shift of DIDS (0.5 mM)-sensitive current induced by a change of [Na+]o corresponded to an apparent coupling ratio of HCO3- to Na+ of 2. RT-PCR analysis showed the presence of transcripts of NBCe1-B, but not NBCe1-A in BPA cells. Electrophysiological and pharmacological properties of whole-cell currents recorded from HEK293 cells transfected with the NBCe1-B, which was cloned from BPA cells resembled those of the native currents. Non-invasive measurements of membrane potential changes in the cell-attached patch configuration indicated that an NBCe activity is present in intact unstimulated BPA cells bathed in a 25 mM HCO3--containing solution. Collectively, these results not only suggest that an NBCe is present, functional and may be mediated, at least in part, by NBCe1-B in BPA cells, but also provide the first electrophysiological characterization of transport properties of NBCe expressed in native exocrine glands. PMID- 16037095 TI - The endocannabinoid system drives neural progenitor proliferation. AB - The discovery of multipotent neural progenitor (NP) cells has provided strong support for the existence of neurogenesis in the adult brain. However, the signals controlling NP proliferation remain elusive. Endocannabinoids, the endogenous counterparts of marijuana-derived cannabinoids, act as neuromodulators via presynaptic CB1 receptors and also control neural cell death and survival. Here we show that progenitor cells express a functional endocannabinoid system that actively regulates cell proliferation both in vitro and in vivo. Specifically, NPs produce endocannabinoids and express the CB1 receptor and the endocannabinoid-inactivating enzyme fatty acid amide hydrolase (FAAH). CB1 receptor activation promotes cell proliferation and neurosphere generation, an action that is abrogated in CB1-deficient NPs. Accordingly, proliferation of hippocampal NPs is increased in FAAH-deficient mice. Our results demonstrate that endocannabinoids constitute a new group of signaling cues that regulate NP proliferation and thus open novel therapeutic avenues for manipulation of NP cell fate in the adult brain. PMID- 16037096 TI - Antibodies against a secreted protein from hookworm larvae reduce the intensity of hookworm infection in humans and vaccinated laboratory animals. AB - The development of a vaccine would provide an important new tool for the control of human hookworm infection. On the basis of successful vaccination of laboratory animals with living irradiated, third-stage hookworm larvae (L3), we examined the antibody responses of individuals from hookworm endemic areas of Brazil and China against the most abundant L3 secreted antigens, the ancylostoma secreted proteins, ASP-1 and ASP-2. Logistic regression was used to investigate the effects of antibody isotype responses to ASPs on the risk of an individual harboring heavy hookworm infection. A significant protective association was observed between increasing anti-ASP-2 IgE levels and the risk of heavy hookworm infection. To confirm that ASP-2 is a protective antigen, laboratory dogs were immunized with recombinant ASP-2 formulated with the GlaxoSmithKline Adjuvant, AS03. Sera obtained from the immunized dogs exhibited high geometric mean antibody titers, immunoprecipitated native ASP-2 from L3 extracts and localized the site of ASP-2 expression to the glandular esophagus and body channels exiting to the cuticle. The sera also exhibited an increased ability to inhibit migration of L3 through tissue in vitro relative to sera from AS03-injected controls. Upon L3 challenge, the ASP-2 vaccinated dogs exhibited significant reductions in fecal egg counts and intestinal hookworm burden. These findings provide strong support for the development of an effective recombinant vaccine against hookworm infection in humans. PMID- 16037097 TI - Haploinsufficiency for trkB and trkC receptors induces cell loss and accumulation of alpha-synuclein in the substantia nigra. AB - The neurotrophins brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT 3) have been shown to promote survival and differentiation of midbrain dopaminergic (DAergic) neurons in vitro and in vivo. This is consistent with their expression and that of their cognate receptors, trkB and trkC, in the nigrostriatal system. Degeneration of DAergic neurons of the substantia nigra and alpha-synuclein-positive aggregates in the remaining substantia nigra (SN) neurons are hallmarks of Parkinson's disease (PD). Reduced expression of BDNF has been reported in the SN from PD patients. Moreover, mutations in the BDNF gene have been found to play a role in the development of familial PD. We show now that haploinsufficiencies of the neurotrophin receptors trkB and/or trkC cause a reduction in numbers of SN neurons in aged (21-23 month old) mice, which is accompanied by a reduced density in striatal tyrosine hydroxylase immunoreactive (TH-ir) fibers. These aged mutant mice, in contrast to wild-type littermates, display an accumulation of alpha-synuclein in the remaining TH-positive neurons of the SN. We conclude that impairment in trkB and/or trkC signaling induces a phenotype in the aged SN, which includes two hallmarks of PD, losses of TH positive neurons and axons along with massive neuronal deposits of alpha synuclein. PMID- 16037098 TI - Identification of CD36 molecular features required for its in vitro angiostatic activity. AB - Thrombospondin-1 (TSP-1), a natural inhibitor of angiogenesis, acts directly on endothelial cells (EC) via CD36 to inhibit their migration and morphogenesis induced by basic fibroblast growth factor. Here we show that CD36 triggered by TSP-1 inhibits in vitro angiogenesis stimulated by vascular endothelial growth factor-A (VEGF-A). To demonstrate that the TSP-1 inhibitory signal was mediated by CD36, we transduced CD36 in CD36-deficient endothelial cells. Both TSP-1 and the agonist anti-CD36 mAb SMO, which mimics TSP-1 activity, reduced the VEGF-A165 induced migration and sprouting of CD36-ECs. To address the mechanisms by which CD36 may exert its angiostatic function, we investigated the functional components of the C-terminal cytoplasmic tail by site-directed mutagenesis. Our results indicate that C464, R467, and K469 of CD36 are required for the inhibitory activity of TSP-1. In contrast, point mutation of C466 did not alter TSP-1 ability to inhibit EC migration and sprouting. Moreover, we show that activation of CD36 by TSP-1 down-modulates the VEGF receptor-2 (VEGFR-2) and p38 mitogen-associated protein kinase phosphorylation induced by VEGF-A165, and this effect was specifically abolished by point mutation at C464. These results identify specific amino acids of the C-terminal cytoplasmic tail of CD36 crucial for the in vitro angiostatic activity of TSP-1 and extend our knowledge of regulation of VEGFR-2-mediated biological activities on ECs. PMID- 16037099 TI - Abrupt reoxygenation following hypoxia reduces electrical coupling between endothelial cells of wild-type but not connexin40 null mice in oxidant- and PKA dependent manner. AB - Although electrical coupling along the arteriolar endothelium is central in arteriolar conducted response and in control of vascular resistance, little is known about the pathophysiological effect of hypoxia and reoxygenation (H/R) on this coupling. We examined this effect in a monolayer of cultured microvascular endothelial cells (ECs) derived from wild-type (WT) or connexin (Cx)40-/- mice (Cx40 is a key gap junction protein in ECs). To assess electrical coupling, we used a current injection technique and Bessel function model to compute the monolayer intercellular resistance. Hypoxia (0.1% O2, 1 h) followed by abrupt reoxygenation (5-90 min) reduced coupling (i.e., increased resistance) in WT but not in Cx40-/- monolayer. H/R increased superoxide production and reduced protein kinase A (PKA) activity in both monolayers. Activation of PKA by 8-bromo-cAMP prevented the reduction in coupling. Preloading of the WT monolayer with the antioxidant ascorbate prevented reductions in both PKA activity and cell coupling. Inhibition of PKA with 6-22 amide during normoxia mimicked the reduction in coupling. Finally, hypoxia followed by slow reoxygenation caused no change in superoxide level, PKA activity, or coupling. Using intravital microscopy, we assessed the physiological relevance of these findings in terms of KCl-induced conducted vasoconstriction in arterioles of WT mouse cremaster muscle in vivo. Ischemia (1 h) followed by abrupt reperfusion (15-30 min) reduced conduction. 8-bromo-cAMP prevented this reduction, while 6-22 amide mimicked this reduction in control nonischemic arterioles. We propose that abrupt reoxygenation reduces interendothelial electrical coupling via oxidant- and PKA-dependent signaling that targets Cx40. We suggest that this mechanism contributes to compromised arteriolar function after H/R. PMID- 16037100 TI - Patient perspectives of patient-provider communication after adverse events. AB - OBJECTIVE: To explore patient perceptions of patient-provider communication after an actual adverse medical event because prior patient error studies are rarely based on real situations. DESIGN: We conducted four patient focus groups using a semi-structured guide. We analyzed transcripts using an editing approach to identify themes. SETTING: Three sites in Colorado. STUDY PARTICIPANTS: participants were recruited from statewide post-injury program. Purposeful sampling began with patients in a geographic location; we contacted every other patient (up to 50). Twenty-two patients initially agreed to participate; 16 adults participated, representing 13 cases. RESULTS: Complex issues and processes were involved in resolution attempts. Effective communication was an important factor in whether professional relationships continued after an adverse event. The communication nature and quality influenced whether patients defined event as 'honest mistake' or 'error'. Two types of trauma (physical and emotional) were expected and found. A third (financial) uncovered and proved in some cases the most salient factor influencing patients' subsequent actions. Caring, honest, quick, personal, and repeated provider responses were linked to patient satisfaction. CONCLUSIONS: Provider communication timeliness and quality were important influences on patients' responses to adverse events. Confronting an adverse medical event collaboratively helped both patients and providers with patients' emotional, physical, and financial trauma and minimized the anger and frustration commonly experienced. Health organizations, providers, investigators, and policymakers should consider the patient experience when developing provider training or evaluating processes in patient resolution. PMID- 16037101 TI - Policymaker use of quality of care information. AB - OBJECTIVE: This study describes public policymakers' experiences with the feasibility of using information from quality of care assessment activities. The objective is to improve the ability to match quality evaluation tools with policymakers' information needs. DESIGN: US state administrative policymakers were interviewed about use of quality of care information and knowledge, attitudes, and experiences with information from specific types of measures. PARTICIPANTS: A purposive sample of 82 key informants from Medicaid program administrations in 48 states. MAIN MEASURE: Users of information from each of eight targeted types of quality of care measurement methods were compared with non-users based upon their levels of knowledge, perceived characteristics of quality of care information, and perceived characteristics of the policy situation. RESULTS: Participants indicated that some types of quality measurement methods have been useful, whereas others have not. Extent of quality assessment information use, and the measurement methods utilized, varied widely. Two factors were associated with the use of information from particular quality assessment methods: information needs of the policymakers and their perceptions of the characteristics, including strengths and weaknesses, of particular measurement methods. CONCLUSIONS: These policymakers had positive attitudes about quality assessment, were knowledgeable about types of methods, and had a variety of potential uses for quality-related information. Yet, perceptions and experiences with different types of measurement methods varied. We describe a set of quality assessment methods with complementary characteristics that could provide a relatively inclusive picture of quality of care and better address policymaker information needs. PMID- 16037102 TI - LDL-cholesterol predicts negative coronary artery remodelling in diabetic patients: an intravascular ultrasound study. AB - AIMS: To investigate the relationship between coronary artery remodelling and glycaemic and lipid profiles in diabetic patients. METHODS AND RESULTS: Intravascular ultrasound analyses of 131 angiographically non-significant coronary stenoses in 80 diabetic patients were performed. The remodelling index (RI) was calculated as the ratio between total vessel area at target site and total vessel area at proximal reference, and was assessed in two ways: as a continuous variable, and as a binary categorical variable: RI<1 namely, negative remodelling (group I), or RI> or =1 (group II). Percentage cross-sectional narrowing was 57+/-13%. On average, RI was 0.93+/-0.13. Coronary shrinkage was found in 94 (71.7%) lesions. Significant inverse correlations were demonstrated between RI and total cholesterol (r=-0.26, P=0.003), apolipoprotein-B (r=-0.23, P=0.01) and LDL-cholesterol (r=-0.3, P=0.001) levels. Multivariable lineal regression analysis identified LDL-cholesterol as the only independent predictor of RI (P=0.001). CONCLUSION: Negative remodelling is a frequent finding in diabetics and it is associated with LDL-cholesterol levels. This may contribute to the diffuse coronary artery disease observed in diabetic patients. PMID- 16037103 TI - The impact of successful assisted reproduction treatment on female and male mental health during transition to parenthood: a prospective controlled study. AB - BACKGROUND: The dynamics of mental health during the transition to parenthood have not been a focus of research. Our prospective longitudinal study was designed to reveal whether there are differences in mental health during the transition to parenthood between parents undergoing treatment with assisted reproduction techniques (ART) and those who conceive spontaneously. METHODS: STUDY GROUP: 367 couples with a singleton ART pregnancy using their own gametes. CONTROL GROUP: 379 couples with a spontaneous singleton pregnancy. Men and women separately filled in questionnaires including the General Health Questionnaire: at the 18th-20th week of pregnancy, 2 months postpartum and 1 year postpartum (T3). The effect of social and child-related factors on mental health was examined. RESULTS: ART women had fewer depressive symptoms during pregnancy than controls, but at T3 their depressive symptoms were at the same level as seen in controls. Anxiety symptoms increased among control but not among ART women across the transition. ART men reported generally fewer mental health symptoms than their controls. Social and child-related stressors had negative impacts on mental health changes among control couples, whereas no impact was found among ART couples. CONCLUSIONS: Successful ART did not predict mental health problems during the transition to parenthood. Moreover, ART couples' mental health was remarkably resistant to social and child-related stress during the transition to parenthood. PMID- 16037104 TI - Alternate day and daily administration of GnRH antagonist may prevent premature luteinization to a similar extent during FSH treatment. AB - BACKGROUND: This randomized controlled trial was designed to evaluate whether a GnRH antagonist given every other day could prevent premature luteinization in women undergoing IVF/ICSI treatment. METHODS: A total of 73 women receiving ovulation stimulation IVF cycles with recombinant FSH were allocated randomly on cycle day 7 to GnRH antagonist ganirelix in multiple doses (0.25 mg each), either daily (n = 37 women, group 1) or every other day (n = 36 women, group 2) until the day of HCG administration. RESULTS: Serum FSH, LH, estradiol and progesterone values showed similar trends in the two groups. During FSH stimulation, 13 (35%) of the women in group 1 had premature LH rises (> or = 10 IU/l) of which eight (22%) were after the start of antagonist administration. In group 2 there were 14 (39%) LH rises during FSH stimulation of which 10 (28%) were after the start of antagonist administration. Luteinization (serum progesterone >2 ng/ml) occurred in only one woman in each group overall (3%). A significantly smaller total dose of the antagonist was used in group 2 than in group 1 (P < 0.001). The study did not have power to evaluate differences in total dose of FSH, number of oocytes recovered and clinical pregnancy rate, all of which appeared similar in the two groups. CONCLUSIONS: Whether alternate day is as effective as daily administration of ganirelix in preventing premature luteinization should be addressed in a non-inferiority trial powered to evaluate live birth rate. PMID- 16037105 TI - Preterm birth in rural Malawi: high incidence in ultrasound-dated population. AB - BACKGROUND: Preterm birth is the major cause of neonatal death, and has an incidence in industrialized countries of 7%. We have found a high incidence (25 30%) previously in a population of anaemic, pregnant women in southern Malawi, studied with ultrasound dating. METHODS: Cohort study of 512 unselected pregnant women in rural communities in Malawi. All had ultrasound fetal measurements before 24 weeks. RESULTS: 20.3% of women delivered before 37 completed weeks of pregnancy. Babies born before 37 completed weeks but after 32 weeks (16%) were twice as likely to die as babies born at term (6.9 versus 3.4%) but this difference did not achieve statistical significance. For those born between 24 and 33 weeks gestation (4.4%) there was a highly significant increase in perinatal mortality (75%) (p <0 .000001). CONCLUSIONS: This population has a very high rate of preterm birth, which is probably infection-related. It may be representative of many rural populations in sub-Saharan Africa. Tackling the problem of neonatal mortality in low income countries will require effective methods to prevent preterm birth. PMID- 16037106 TI - Polymorphisms in the insulin receptor substrate-1 (IRS-1) gene and the insulin receptor substrate-2 (IRS-2) gene influence glucose homeostasis and body mass index in women with polycystic ovary syndrome and non-hyperandrogenic controls. AB - BACKGROUND: We aimed to evaluate the influence of the Gly972Arg variant of the insulin receptor substrate-1 gene (IRS-1) and the Gly1057Asp variant in IRS-2 on insulin resistance and glucose tolerance in women with polycystic ovary syndrome (PCOS) and healthy controls. METHODS: Genotypes, allelic frequencies, indexes of insulin resistance, glucose tolerance and hormone profiles were studied in a large sample of Spanish PCOS (n = 103) women compared with a control group (n = 48) of healthy women matched for body mass index. RESULTS: No differences in genotype or allelic frequencies were found between PCOS patients and healthy controls. When considering control subjects and PCOS patients as a whole, IRS-1 Arg972 carriers also presented with increased fasting insulin (133 +/- 60 versus 95 +/- 67 pmol/l, P = 0.008) and insulin resistance measured by homeostasis model assessment (4.3 +/- 2.1 versus 3.1 +/- 2.4, P = 0.009) compared with subjects homozygous for Gly972 alleles. These differences were even higher when restricting the analysis to PCOS patients. Subjects homozygous for the Gly1057 allele of IRS-2 presented with increased 60 and 90 min oral glucose tolerance test (OGTT) glucose levels compared with carriers of one or two Asp1057 alleles (7.9 +/- 2.1 versus 7.1 +/- 2.1 mmol/l, P = 0.042 and 7.0 +/- 2.1 versus 6.0 +/- 1.8 mmol/l, P = 0.014), and a similar tendency was observed for 120 min OGTT glucose levels. CONCLUSIONS: The Gly972Arg in IRS-1 and Gly1057Asp in IRS-2 polymorphisms influence glucose homeostasis in premenopausal women, but are not associated with PCOS. PMID- 16037107 TI - Misoprostol moistened with acetic acid or saline for second trimester pregnancy termination: a randomized prospective double-blind trial. AB - BACKGROUND: This study was conducted to evaluate the efficacy and side effects of a new regimen of 800 microg misoprostol administered intravaginally every 6 h up to a maximum of three doses in 24 h for second trimester pregnancy termination. METHODS: A total of 66 women seeking termination of second trimester pregnancy (30 fetal structural anomaly, six chromosomal abnormality and 30 fetal death) were randomly assigned to one of two treatment groups: (i) intravaginal misoprostol moistened with 3 ml of 5% acetic acid in group A (n = 33); or (ii) intravaginal misoprostol moistened with 3 ml of saline in group B (n = 33). RESULTS: The overall median (range) induction-abortion interval was 10 h (2-46) [10 h (4-35) in 36 live fetuses and 9 h (2-46) in 30 dead fetuses, P = 0.515]. All of the patients in both groups aborted within 48 h (100% success rate). The median (range) induction-abortion interval revealed a significantly faster delivery time (P < 0.001) in group A [8 h (2-24)] than in group B [14 h (3-46)]. CONCLUSIONS: This new regimen of 800 microg of vaginal misoprostol every 6 h for a maximum of three doses in 24 h was an effective alternative method for second trimester abortion. In addition, misoprostol moistened with acetic acid was significantly more effective than misoprostol moistened with saline. PMID- 16037108 TI - Towards less confusing terminology in reproductive medicine: clarifying medical ambiguities to the benefit of all. AB - It has been argued that terminology in reproductive medicine related to reproductive success is ambiguous, confusing and misleading. A proposed solution is the conditional use of the term 'fertility' which is qualified according to statements concerning description, diagnosis and prognosis, and for which a grading system is proposed. We argue that there already exists (from 1989) a well articulated conceptualization of fertility that does not appear to have been well disseminated within reproductive medicine. Within this conceptualization there is an important place for separate terms that clearly distinguish aetiology from outcome. We therefore see a central role for maintaining and articulating the terms fecundity and fertility. It is also suggested that one source of confusion within reproductive medicine stems from the conflation of interests for clinicians and patients when discussing infertility as both a cause and an outcome. Unpacking the meaning of the term 'infertility' reveals a complex of interdependent concepts that are both social and biological in origin. PMID- 16037109 TI - Microvascular dysfunction in women with polycystic ovary syndrome. AB - BACKGROUND: Polycystic ovary syndrome (PCOS) is associated with multiple cardiovascular risk factors and an increased prevalence of arterial dysfunction. However, microvascular dysfunction in PCOS has not been assessed. METHODS: Subjects comprised 12 women with PCOS and 12 age-matched controls with normal ovaries. Microvascular function was assessed by observing forearm skin microvascular erythrocyte flux responses, to cumulative iontophoretic doses of 1% (w/v) acetylcholine (ACh) and 1% (w/v) sodium nitroprusside (SNP), using laser Doppler imaging. RESULTS: Basal microvascular perfusion was comparable in PCOS and controls. The increase in skin microvascular perfusion in response to ACh was however generally blunted in PCOS women (P = 0.018). Peak ACh-induced erythrocyte flux was also less (p < 0.04) in PCOS women (125.1 +/- 21.7, i.e. 5.3-fold basal flux) than in controls (200.8 +/- 28.5, i.e. 8.3-fold basal flux). Analysis of covariance indicated this effect was unrelated to differences in body mass index or serum testosterone but serum insulin may be a weak confounder. No differences were noted between the PCOS and control groups in their response to SNP. CONCLUSION: Despite its limited sample size studied, this is the first demonstration that women with PCOS exhibit microvascular endothelial dysfunction, indicated by an inhibited vasodilatory response to ACh. PMID- 16037110 TI - A prospective randomized comparison of sublingual and oral misoprostol when combined with mifepristone for medical abortion at 12-20 weeks gestation. AB - BACKGROUND: Sublingual misoprostol has been shown to be effective in medical abortion. A prospective double-blinded placebo-controlled trial was done to compare the efficacy and side-effects of sublingual to oral misoprostol when used with mifepristone for medical abortion from 12 to 20 weeks gestation. METHODS: A total of 120 women at 12-20 weeks of gestation were randomized to receive 200 mg oral mifepristone followed by either sublingual or oral misoprostol 400 mg every 3 h for a maximum of five doses 36-48 h later. The course of misoprostol was repeated if the woman did not abort within 24 h. RESULTS: There was no significant difference (P = 0.43) in the success rate at 24 h [relative risk = 1.075; 95% confidence interval (CI): 0.94-1.19]. Abortion occurred in 91.4% in the sublingual group (95% CI: 81.0-96.7%) as compared to 85.0% (95% CI: 73.7 92.1%) in the oral group. The median induction-to-abortion interval was significantly shorter (P = 0.009) in the sublingual group (5.5 h) as compared to the oral group (7.5 h). The incidence of fever was higher in the sublingual group (P < 0.0001). The incidences of other side-effects were similar. CONCLUSION: Sublingual misoprostol, when combined with mifepristone, is effective for medical abortion in the second trimester. The induction-to-abortion interval is shorter when sublingual misoprostol is used when compared to oral misoprostol. PMID- 16037111 TI - GnRH II as a possible cytostatic regulator in the development of endometriosis. AB - BACKGROUND: GnRH II is the second form of GnRH and is widely distributed in peripheral tissues of the female reproductive tract as well as in the central nervous system. In the present study, we studied the possible implication of GnRH II in endometriosis. METHODS: Effects of GnRH II on 5-bromo-2'-deoxyuridine (BrdU) uptake by cultured endometriotic stromal cells were examined. Effects of GnRH II on interleukin (IL)-1beta-induced expression of cyclooxygenase (COX)-2 and IL-8 were also studied. mRNA levels of GnRH I, GnRH II, type I GnRH receptor and type II GnRH receptor were determined by real-time quantitative RT-PCR in endometrial tissues of women with or without endometriosis and in endometriotic tissues. RESULTS: GnRH II dose-dependently suppressed BrdU uptake by endometrial stromal cells. Treatment with IL-1beta markedly increased mRNA levels of COX-2 and IL-8 in endometrial stromal cells and IL-8 protein secretion by these cells, while these increments were significantly suppressed by supplementation with GnRH II. The mRNA levels of GnRH II were lower in endometrial and endometriotic tissues of women with endometriosis than in endometrial tissues of women without endometriosis, both in the proliferative phase and the secretory phase. In addition, as for GnRH I, type I GnRH receptor and type II GnRH receptor, the mRNA levels were lower in endometrial tissues of women with endometriosis than in those without endometriosis in the secretory phase. CONCLUSIONS: In the light of the demonstrated antiproliferative and anti-inflammatory effects of GnRH II on endometrial stromal cells, the lower expression of GnRH II in eutopic and ectopic endometrium of women with endometriosis suggests that endogenous GnRH II-mediated cytostatic regulation may be impaired in the development of endometriosis. PMID- 16037112 TI - The effect of endometriosis, cycle stage, lymphocyte suppression and pregnancy on CA-125 levels in peritoneal fluid and serum in baboons. AB - BACKGROUND: Serum CA-125 during the mid-follicular phase has been reported to be a clinically useful and reproducible marker in the diagnosis of advanced endometriosis in women. This study was undertaken to document the effect of the menstrual cycle, pregnancy and lymphocyte suppression on CA-125 levels in peritoneal fluid (PF) and serum in baboons with a normal pelvis and baboons with endometriosis. METHODS: CA-125 levels were measured in 264 serum samples that were serially obtained during one menstrual cycle from 10 animals with and without endometriosis. In addition, CA-125 levels were determined in 204 archived samples (serum, n = 112 and PF, n = 92) obtained from 32 female baboons with or without endometriosis. The CA-125 assays were performed by radioimmunoassay using kits from Centocor (Malvern, PA, USA). RESULTS: Serum CA-125 levels were at their highest during menstruation and decreased progressively during the follicular and luteal phase. PF CA-125 levels were increased during the follicular phase in baboons with a normal pelvis, but no cyclic changes were observed in animals with endometriosis. Serum CA-125 levels were unaffected by induction, lymphocyte suppression or pregnancy. Induction of endometriosis resulted in increased PF CA 125 levels, whereas lymphocyte suppression or pregnancy had no effect. CONCLUSION: In baboons, serum CA-125 originates mainly from eutopic endometrium whereas the main source of PF CA-125 seems to be the peritoneum or ectopic endometrium. The baboon appears to be a valid model to further study the relationship between endometriosis and CA-125. PMID- 16037113 TI - Further considerations on natural or mild hyperstimulation cycles for intrauterine insemination treatment: effects on pregnancy and multiple pregnancy rates. AB - BACKGROUND: The high iatrogenic multiple pregnancy rate associated with intrauterine insemination (IUI) in hyperstimulated cycles is becoming less acceptable. Therefore we investigated data from an earlier prospective trial with regard to the specific question of whether the application of mild hyperstimulation in IUI cycles could be an alternative strategy for obtaining acceptable pregnancy rates while preventing a high multiple pregnancy rate, compared with natural cycles for IUI. METHODS: Pregnancy outcome of 310 natural and 334 mildly hyperstimulated cycles for IUI in 171 couples with unexplained or mild male factor subfertility was analysed on a patient level with random coefficient models. RESULTS: Pregnancy rates were similar: 35 and 39.8% per couple in the natural and mildly hyperstimulated cycles respectively (P = 0.60). Multiple pregnancies, all twin pregnancies, were conceived significantly more frequently in the mild hyperstimulation group (27% of the pregnancies) than in the natural cycle group (4% of the pregnancies) (P = 0.01). All multiple pregnancies in the hyperstimulation group were conceived in multifollicular cycles. Multifollicular development was strongly associated with the application of mild hyperstimulation only (odds ratio 21.14, 95% confidence interval 8.15 54.79). CONCLUSION: The application of a mild hyperstimulation protocol as an alternative to a standard hyperstimulation protocol for IUI does not result in higher pregnancy rates than IUI in the natural cycle, while at the same time multiple pregnancies cannot be avoided. Therefore, there is no place for the use of gonadotrophins in IUI treatment. PMID- 16037114 TI - Decline in fertility of mouse sperm with abnormal chromatin during epididymal passage as revealed by ICSI. AB - BACKGROUND: Recent studies showed that ICSI with cauda epididymal or ejaculated sperm of infertile mice or men, respectively, was less effective in fertilization and normal embryo development than ICSI using sperm from the testes. These studies suggested that sperm nuclear quality declined after release from the testis, but the site where this loss of fertility occurs has not been localized. METHODS: We performed ICSI with testicular, caput, and cauda epididymal sperm from infertile Tnp1-/-Tnp2+/- mutant mice, which have a minimal level of transition nuclear proteins and are sterile by natural mating. RESULTS: When the heads of motile sperm from the testis or caput epididymis of Tnp1-/-Tnp2+/- males were injected into enucleated mouse oocytes, sperm chromosomes showed no difference from those of wild-type mice, but the chromosomes from sperm taken from the cauda epididymis of mutant males showed increased abnormalities. Injection of testicular or caput epididymal sperm from Tnp1-/-Tnp2+/- males into intact oocytes resulted in normal embryonic and fetal development and yields of liveborn equivalent to wild-type, but cauda sperm from Tnp1-/-Tnp2-/- mice produced lower implantation rates and yields of liveborn than did those from wild type mice. CONCLUSIONS: These results demonstrate that in mice with sperm chromatin abnormalities, the decline in fertility of sperm with ICSI occurs after the caput epididymis. The advantage of using caput epididymal sperm for ICSI in certain situations may be considered as an approach to be tested in human assisted reproduction. PMID- 16037115 TI - Nucleocytoplasmic ratio of fully grown germinal vesicle oocytes is essential for mouse meiotic chromosome segregation and alignment, spindle shape and early embryonic development. AB - BACKGROUND: This study examined the effect of nucleocytoplasmic ratio of fully grown germinal vesicle (GV) oocytes on meiotic chromosome segregation and alignment, spindle shape, Ca(2+) oscillations and capacity of early embryonic development in mouse. METHODS: GV oocytes with reduced volume (equal to 1/5 to 4/5 of an intact oocyte) were made by micromanipulation to remove different amounts of cytoplasm, and then matured and fertilized in vitro. RESULTS: When >1/2 of GV oocyte cytoplasm was removed, the time-course of GV breakdown (GVBD) was delayed and oocyte maturation rate decreased significantly. Abnormal chromosome segregation rate increased if >1/2 of the cytoplasm was removed from the oocyte. Length and structure of meiotic spindle and chromosome alignment were also impaired by the reduction of cytoplasmic volume. Once matured in vitro, the oocytes could undergo Sr(2+)-induced Ca(2+) oscillations and form pronuclei in a manner independent of nucleocytoplasmic ratio, but their ability to develop to 2 cell embryos was affected if >1/2 of their cytoplasm was removed from the GV oocytes. CONCLUSIONS: These results suggest that nucleocytoplasmic ratio is essential for normal meiotic chromosome segregation, spindle formation and chromosome alignment over the metaphase spindle, and development to 2-cell stage, for which 1/2 of the volume of the GV oocyte appears to be a threshold. PMID- 16037116 TI - The implantation of every embryo facilitates the chances of the remaining embryos to implant in an IVF programme: a mathematical model to predict pregnancy and multiple pregnancy rates. AB - BACKGROUND: We aimed to assess the validity of a theoretical mathematical model to predict the pregnancy rate and the multiple pregnancy rate in IVF/oocyte donation programmes on the basis of the implantation rate and the number of transferred embryos. METHODS: A total of 1835 embryo transfers corresponding to three different programmes in two centres with different implantation rates were analysed. Pregnancy and multiple pregnancy rates observed in the aforementioned programmes were compared with those obtained following different mathematical models. Four models were tested: binomial model, ground model, maternal variability model and collaborative model. The goodness of fit was performed by means of the maximum likelihood fit method. RESULTS: The binomial model could not predict the pregnancy rate, and especially the multiple pregnancy rate. The multiple pregnancy rate predicted following the binomial model was much lower than observed, up to 40-fold reduced. Ground model and maternal variability model adjusted to the data with more precision, but were still not accurate. Finally, the collaborative model reproduced with very great accuracy both pregnancy rate and the multiple pregnancy rate. A collaborative parameter of 22% was found, implying that the implantation probability of each embryo is increased by 22% for every embryo previously implanted. CONCLUSIONS: Embryonic implantation does not follow a binomial law, showing that the implantation is not independent from the number of embryos implanted. The best fit to the data is obtained following a collaborative model by which the implantation of one embryo is facilitated by the implantation of other embryo(s). The mathematical formula of the collaborative model predicts very accurately the pregnancy rate and the multiple pregnancy rate in IVF/oocyte donation programmes, based on the implantation rate of this specific programme and the number of embryos transferred up to five embryos. We recommend using the aforementioned formula to quantify the pregnancy rate and the risk of multiple pregnancy in the counselling of the infertile couple at embryo transfer. Such a formula is freely available at www.ifca.unican.es/matorras/mathpreg/. PMID- 16037117 TI - Fertilization, cleavage and blastocyst development according to the maturation timing of oocytes in in vitro maturation cycles. AB - BACKGROUND: This study was to examine the developmental capacity of oocytes collected from an in vitro maturation (IVM) programme according to their maturation time. METHODS: The study included 47 IVM cycles that underwent blastocyst transfer. The patients (n = 38) were primed with 10 000 IU HCG 36 h before their oocyte retrieval. The oocytes were classified into three groups: group 1 (n = 139) where oocytes were matured on day of oocyte collection; group 2 (n = 627) where oocytes were matured on day 1 after IVM; group 3 (n = 163) where oocytes matured on day 2 after IVM. Fertilization, cleavage and blastocyst formation were compared between three groups. RESULTS: Rates of cleavage and blastocyst development in group 3 (72.2%, 96/133; 19.0%, 15/133) were significantly lower than those of group 1 (100%, 108/108; 58.3%, 63/108) and group 2 (91.5%, 487/532; 50.4%, 268/532) respectively (P < 0.01). The number of freezable good quality blastocysts among blastocysts developed from group 1 (52.4%, 33/63) was significantly higher than those from group 2 (35.4%, 95/268) and group 3 (6.7%, 1/15) (P < 0.01). There were 24 clinical pregnancies (51.1%, 24/47) after transfer of the blastocysts and 29 healthy babies were delivered. CONCLUSION: These results suggest that oocytes reaching metaphase II faster in an IVM programme have better embryonic developmental competence. PMID- 16037118 TI - Is there a relation between school smoking policies and youth cigarette smoking knowledge and behaviors? AB - To comply with workplace legislation, New Zealand schools are required to have policies regarding tobacco smoking. Many schools also have policies to prevent tobacco use by students, including education programmes, cessation support and punishment for students found smoking. This paper investigated the associations between school policies and the prevalence of students' cigarette smoking. Furthermore, we investigated the association between school policy and students' tobacco purchasing behavior, knowledge of health effects from tobacco use and likelihood of influencing others not to smoke. Data were obtained from a self report survey administered to 2,658 New Zealand secondary school students and staff from 63 schools selected using a multi-stage sampling procedure. Components of school policy were not significantly associated with smoking outcomes, health knowledge or health behavior, and weakly related to a punishment emphasis and students advising others to not smoke. Similarly, weak associations were found between not advising others to not smoke and policies with a punishment emphasis as well as smoke-free environments. The results suggest that having a school tobacco policy was unrelated to the prevalence of tobacco use among students, tobacco purchasing behavior and knowledge of the negative health effects of tobacco. PMID- 16037119 TI - Genetic polymorphisms and the effect of cigarette smoking in the comet assay. AB - A potential genotoxic effect of cigarette smoking has repeatedly been investigated with the comet assay (alkaline single cell gel electrophoresis) and conflicting results have been reported. Besides differences in the methodology and the study design used, genetic differences between the subjects investigated might contribute to the variability of test results. Considering genetic polymorphisms of genes involved in metabolism or DNA repair has led to a better discrimination of smoking-related genotoxic effects in some cases but also led to discrepant results. We therefore evaluated our baseline comet assay effects obtained for nonsmokers and smokers in relation to selected genetic polymorphisms. Our study group comprised 52 nonsmokers and 51 smokers who were strictly selected to exclude potential confounding factors. We chose polymorphisms in the genes GSTM1 and CYP1A1 (Ile462Val) because they take part in the metabolism of genotoxins contained in tobacco smoke. In a subgroup of 32 nonsmokers and 31 smokers we also studied polymorphisms in XPD (Lys751Gln), XRCC1 (Arg399Gln) and XRCC3 (Thr241Val) because they are part of DNA repair pathways involved in the repair of tobacco-related DNA damage. Freshly collected peripheral whole blood samples were tested in the alkaline (pH > 13) comet assay. In all experiments a reference standard (untreated V79 cells) was included to correct for assay variability. An independent second evaluation was carried out for all experiments. None of these approaches revealed a significant difference between nonsmokers and smokers. PMID- 16037120 TI - Investigating genetic damage in workers occupationally exposed to methotrexate using three genetic end-points. AB - Genetic damage in workers occupationally exposed to an antineoplastic drug was studied using the micronucleus (MN) test, the comet assay, the hprt gene mutation assay and the TCR gene mutation assay. The subjects were divided into two groups: (i) 21 workers from a plant producing methotrexate (MTX); (ii) 21 controls were matched according to age, gender and smoking. Fresh blood samples were collected from the workers and controls. The results of the MN test showed that the mean micronuclei rate (MNR) and mean micronucleated cell rate (MCR) in workers were 10.10 +/- 0.95 per thousand and 8.05 +/- 0.75 per thousand, respectively, which were significantly higher than those (5.48 +/- 0.82 per thousand and 4.38 +/- 0.58 per thousand) in controls (P < 0.01). It was found in the comet assay that the mean tail length (MTL) of workers and controls were 1.30 +/- 0.06 microm and 0.07 +/- 0.01 microm, respectively. There was a significant difference between workers and controls for MTL (P < 0.01), but the difference between the mean tail moment (MTM, 0.23 +/- 0.03) of workers and MTM (0.17 +/- 0.04) of controls was not significant (P > 0.05). The results of hprt gene mutation assay showed that the average mutation frequency (Mf-hprt) of hprt in workers was 1.00 +/- 0.02 per thousand, which was significantly higher than that (0.86 +/- 0.01 per thousand) in controls (P < 0.01). Meanwhile, the results of TCR gene mutation assay indicated that Mfs-TCR gene mutation frequencies of workers and controls were 6.87 +/- 0.52 x 10(-4) and 1.67 +/- 0.14 x 10(-4), respectively, which were significantly different (P < 0.01). The results of our experiment suggest that genetic damage is detectable in the 21 workers occupationally exposed to methotrexate. PMID- 16037121 TI - Resolving abbreviations to their senses in Medline. AB - MOTIVATION: Biological literature contains many abbreviations with one particular sense in each document. However, most abbreviations do not have a unique sense across the literature. Furthermore, many documents do not contain the long forms of the abbreviations. Resolving an abbreviation in a document consists of retrieving its sense in use. Abbreviation resolution improves accuracy of document retrieval engines and of information extraction systems. RESULTS: We combine an automatic analysis of Medline abstracts and linguistic methods to build a dictionary of abbreviation/sense pairs. The dictionary is used for the resolution of abbreviations occurring with their long forms. Ambiguous global abbreviations are resolved using support vector machines that have been trained on the context of each instance of the abbreviation/sense pairs, previously extracted for the dictionary set-up. The system disambiguates abbreviations with a precision of 98.9% for a recall of 98.2% (98.5% accuracy). This performance is superior in comparison with previously reported research work. AVAILABILITY: The abbreviation resolution module is available at http://www.ebi.ac.uk/Rebholz/software.html. PMID- 16037122 TI - Lactate is a critical "sensed" variable in caudal hindbrain monitoring of CNS metabolic stasis. AB - Caudal hindbrain "sensing" of glucoprivation activates central neural mechanisms that enhance systemic glucose availability, but the critical molecular variable(s) linked to detection of local metabolic insufficiency remains unclear. Central neurons and glia are metabolically coupled via intercellular trafficking of the glycolytic product lactate as a substrate for neuronal oxidative respiration. Using complementary in vivo models for experimental manipulation of lactate availability within the caudal hindbrain, we investigated the hypothesis that lactate insufficiency may be monitored by local metabolically "sensitive" neurons as an indicator of central nervous system energy imbalance. The data show that caudal fourth ventricular (CV4) administration of the monocarboxylate transporter inhibitor alpha-cyano-4-hydroxycinnamate (4CIN) resulted in dose dependent increases in blood glucose in euglycemic animals, whereas the degree and duration of hypoglycemia elicited by insulin administration were exacerbated by exogenous L-lactate delivery to the CV4. Immunocytochemical processing of the hindbrain for the inducible c-fos gene product Fos revealed that 4CIN enhanced Fos immunoreactivity in the dorsal vagal complex (DVC), e.g., the nucleus of the solitary tract and dorsal vagal motor nucleus, and adjacent area postrema, sites where cells characterized by unique sensitivity to diminished glucose and/or glycolytic intermediate/end product levels reside, and in the medial vestibular nucleus (MV), and that CV4 L-lactate infusion increased Fos labeling within the DVC and MV after insulin-induced hypoglycemia. Together, these results support the view that lactate is a critical monitored metabolic variable in caudal hindbrain detection of energy imbalance resulting from glucoprivation and that diminished uptake and/or oxidative catabolism of this fuel activates neural mechanisms that increase systemic glucose availability. PMID- 16037123 TI - Microvascular rarefaction and decreased angiogenesis in rats with fetal programming of hypertension associated with exposure to a low-protein diet in utero. AB - In hypertension, increased peripheral vascular resistance results from vascular dysfunction with or without structural changes (vessel wall remodeling and/or microvascular rarefaction). Humans with lower birth weight exhibit evidence of vascular dysfunction. The current studies were undertaken to investigate whether in utero programming of hypertension is associated with in vivo altered response and/or abnormal vascular structure. Offspring of Wistar dams fed a normal (CTRL) or low (LP)-protein diet during gestation were studied. Mean arterial blood pressure response to ANG II was significantly increased, and depressor response to sodium nitroprusside (SNP) infusions significantly decreased in male LP adult offspring relative to CTRL. No arterial remodeling was observed in male LP compared with CTRL offspring. Capillary and arteriolar density was significantly decreased in striated muscles from LP offspring at 7 and 28 days of life but was not different in late fetal life [day 21 of gestation (E21)]. Angiogenic potential of aortic rings from LP newborn (day of birth, P0) was significantly decreased. Striated muscle expressions (Western blots) of ANG II AT(1) receptor subtype, endothelial nitric oxide synthase, angiopoietin 1 and 2, Tie 2 receptor, vascular endothelial growth factor and receptor, and platelet-derived growth factor C at E21 and P7 were unaltered by antenatal diet exposure. In conclusion, blood pressure responses to ANG II and SNP are altered, and microvascular structural changes prevail in this model of fetal programming of hypertension. The capillary rarefaction is absent in the fetus and appears in the neonatal period, in association with decreased angiogenic potential. The study suggests that intrauterine protein restriction increases susceptibility to postnatal factors resulting in microvascular rarefaction, which could represent a primary event in the genesis of hypertension. PMID- 16037124 TI - Dipalmitoylphosphatidylcholine is not the major surfactant phospholipid species in all mammals. AB - Pulmonary surfactant, a complex mixture of lipids and proteins, lowers the surface tension in terminal air spaces and is crucial for lung function. Within an animal species, surfactant composition can be influenced by development, disease, respiratory rate, and/or body temperature. Here, we analyzed the composition of surfactant in three heterothermic mammals (dunnart, bat, squirrel), displaying different torpor patterns, to determine: 1) whether increases in surfactant cholesterol (Chol) and phospholipid (PL) saturation occur during long-term torpor in squirrels, as in bats and dunnarts; 2) whether surfactant proteins change during torpor; and 3) whether PL molecular species (molsp) composition is altered. In addition, we analyzed the molsp composition of a further nine mammals (including placental/marsupial and hetero-/homeothermic contrasts) to determine whether phylogeny or thermal behavior determines molsp composition in mammals. We discovered that like bats and dunnarts, surfactant Chol increases during torpor in squirrels. However, changes in PL saturation during torpor may not be universal. Torpor was accompanied by a decrease in surfactant protein A in dunnarts and squirrels, but not in bats, whereas surfactant protein B did not change in any species. Phosphatidylcholine (PC)16:0/16:0 is highly variable between mammals and is not the major PL in the wombat, dunnart, shrew, or Tasmanian devil. An inverse relationship exists between PC16:0/16:0 and two of the major fluidizing components, PC16:0/16:1 and PC16:0/14:0. The PL molsp profile of an animal species is not determined by phylogeny or thermal behavior. We conclude that there is no single PL molsp composition that functions optimally in all mammals; rather, surfactant from each animal is unique and tailored to the biology of that animal. PMID- 16037125 TI - Lipopolysaccharide-induced fever in Pekin ducks is mediated by prostaglandins and nitric oxide and modulated by adrenocortical hormones. AB - Information on avian fever is limited, and, in particular, very little is known about the mediators and modulators of the febrile response in birds. Therefore, in this study, the possible mediatory roles of nitric oxide (NO) and prostaglandins (PGs), together with a potential modulatory role for adrenocortical hormones in the generation of fever was investigated in conscious Pekin ducks. Their body temperatures were continuously measured by abdominally implanted temperature-sensitive data loggers. The febrile response induced by intramuscular injection of LPS at a dose of 100 microg/kg was compared with and without inhibition of NO production by N-nitro-L-arginine methyl ester (L-NAME), inhibition of PG synthesis (by diclofenac), and elevation of circulating concentrations of dexamethasone and corticosterone (by exogenous administration). LPS administration induced a marked, monophasic fever with a rise in temperature of more than 1 degrees C after 3-4 h. In the presence of L-NAME, diclofenac, and adrenocorticoids at doses that had no effect upon normal body temperature in afebrile ducks, there was a significant inhibition of the LPS-induced fever. In addition, during the febrile response, the blood concentration of corticosterone was significantly elevated (from a basal level of 73.6 +/- 9.8 ng/ml to a peak level of 132.6 +/- 16.5 ng/ml). The results strongly suggest that the synthesis of both NO and PGs is a vital step in the generation of fever in birds and that the magnitude of the response is subject to modulation by adrenocorticoids. PMID- 16037126 TI - Fever suppression in near-term pregnant rats is dissociated from LPS-activated signaling pathways. AB - Near-term pregnant rats show a suppressed fever response to LPS that is associated with reduced induction of cyclooxygenase (COX)-2 in the hypothalamus. The objective of this study is to explore whether the LPS-activated signaling pathways in the fever-controlling region of the hypothalamus are specifically altered at near term. Three rat groups consisting of 15-day pregnant rats, near term 21- to 22-day pregnant rats, and day 5 lactating rats were injected with a febrile dose of LPS (50 mug/kg ip). The hypothalamic preoptic area and the organum vasculosum of the lamina terminalis (OVLT) were collected 2 h after LPS injection. The activation of three transcription modulators, nuclear factor kappaB (NF-kappaB), extracellular signal-regulated kinase 1/2 (ERK1/2), and signal transducer and activator of transcription 5 (STAT5), was assessed using semiquantitative Western blot analysis. LPS activated the NF-kappaB pathway in all rat groups, and this response was not altered at near term. ERK1/2 and STAT5 were constitutively activated during all reproductive stages, and their levels were not significantly affected by LPS injection. Plasma levels of the proinflammatory cytokines (IL-1beta, IL-6, TNF-alpha, and IFN-gamma), anti inflammatory cytokines (IL-4, IL-10, and IL-1 receptor antagonist), and corticosterone were unaffected during the three reproductive stages after LPS challenge. We observed a sharp decrease in the expression of a prostaglandin producing enzyme called lipocalin-prostaglandin D2 synthase in near-term pregnant and lactating rats. Thus fever suppression at near term is not due to an alteration in either LPS-activated intracellular signaling pathways or LPS induced pro- and anti-inflammatory cytokine production. PMID- 16037127 TI - Cardiovascular responses to water drinking: does osmolality play a role? AB - Water drinking activates the autonomic nervous system and induces acute hemodynamic changes. The actual stimulus for these effects is undetermined but might be related to either gastric distension or to osmotic factors. In the present study, we tested whether the cardiovascular responses to water drinking are related to water's relative hypoosmolality. Therefore, we compared the cardiovascular effects of a water drink (7.5 ml/kg body wt) with an identical volume of a physiological (0.9%) saline solution in nine healthy subjects (6 male, 3 female, aged 26 +/- 2 years), while continuously monitoring beat-to-beat blood pressure (finger plethysmography), cardiac intervals (electrocardiography), and cardiac output (thoracic impedance). Total peripheral resistance was calculated as mean blood pressure/cardiac output. Cardiac interval variability (high-frequency power) was assessed by spectral analysis as an index of cardiac vagal tone. Baroreceptor sensitivity was evaluated using the sequence technique. Drinking water, but not saline, decreased heart rate (P = 0.01) and increased total peripheral resistance (P < 0.01), high-frequency cardiac interval variability (P = 0.03), and baroreceptor sensitivity (P = 0.01). Neither water nor saline substantially increased blood pressure. These responses suggest that water drinking simultaneously increases sympathetic vasoconstrictor activity and cardiac vagal tone. That these effects were absent after drinking physiological saline indicate that the cardiovascular responses to water drinking are influenced by its hypoosmotic properties. PMID- 16037128 TI - Negative regulation of growth hormone receptor signaling. AB - GH has been of significant scientific interest for decades because of its capacity to dramatically change physiological growth parameters. Furthermore, GH interacts with a range of other hormonal pathways and is an established pharmacological agent for which novel therapeutical applications can be foreseen. It is easy to see the requirement for a number of postreceptor mechanisms to regulate and control target tissue sensitivity to this versatile hormone. In recent years, some of the components that take part in the down-regulatory mechanism targeting the activated GH receptor (GHR) have been defined, and the physiological significance of some of these key components has begun to be characterized. Down-regulation of the GHR is achieved through a complex mechanism that involves rapid ubiquitin-dependent endocytosis of the receptor, the action of tyrosine phosphatases, and the degradation by the proteasome. The suppressors of cytokine signaling (SOCS) protein family, particularly SOCS2, plays an important role in regulating GH actions. The aim of this review is to summarize collected knowledge, including very recent findings, regarding the intracellular mechanisms responsible for the GHR signaling down-regulation. Insights into these mechanisms can be of relevance to several aspects of GH research. It can help to understand growth-related disease conditions, to explain GH resistance, and may be used to develop pharmaceuticals that enhance some the beneficial actions of endogenously secreted GH in a tissue-specific manner. PMID- 16037129 TI - New modes of action for endocrine-disrupting chemicals. AB - Endocrine-disrupting chemicals (EDC) are commonly considered to be compounds that mimic or block the transcriptional activation elicited by naturally circulating steroid hormones by binding to steroid hormone receptors. For example, the Food Quality Protection Act of 1996 defines EDC as those, that "may have an effect in humans that is similar to an effect produced by a naturally occurring estrogen, or other such endocrine effect as the Administrator may designate." The definition of EDC was later expanded to include those that act on the estrogen, androgen, and thyroid hormone receptors. In this minireview, we discuss new avenues through which xenobiotic chemicals influence these and other hormone dependent signaling pathways. EDC can increase or block the metabolism of naturally occurring steroid hormones and other xenobiotic chemicals by activating or antagonizing nuclear hormone receptors. EDC affect the transcriptional activity of nuclear receptors by modulating proteasome-mediated degradation of nuclear receptors and their coregulators. Xenobiotics and environmental contaminants can act as hormone sensitizers by inhibiting histone deacetylase activity and stimulating mitogen-activated protein kinase activity. Some endocrine disrupters can have genome-wide effects on DNA methylation status. Others can modulate lipid metabolism and adipogenesis, perhaps contributing to the current epidemic of obesity. Additional elucidation of these new modes of endocrine disruption will be key in understanding the nature of xenobiotic effects on the endocrine system. PMID- 16037130 TI - The glucose-6-phosphatase catalytic subunit gene promoter contains both positive and negative glucocorticoid response elements. AB - Glucose-6-phosphatase catalyzes the final step in the gluconeogenic and glycogenolytic pathways. Glucocorticoids stimulate glucose-6-phosphatase catalytic subunit (G6Pase) gene transcription and studies performed in H4IIE hepatoma cells demonstrate the presence of a glucocorticoid response unit (GRU) in the proximal G6Pase promoter. In vitro deoxyribonuclease I footprinting analyses show that the glucocorticoid receptor binds to three glucocorticoid response elements (GREs) in the -231 to -129 promoter region and transfection results indicate all three contribute to glucocorticoid induction of G6Pase gene transcription. Furthermore, binding sites for hepatocyte nuclear factor-1 and -4, CRE binding factors, and FKHR (FOXO1a) are required for the full glucocorticoid response. Chromatin immunoprecipitation assays show that dexamethasone treatment stimulates glucocorticoid receptor and FKHR binding to the endogenous G6Pase promoter. Surprisingly, although glucocorticoids stimulate G6Pase gene transcription, deoxyribonuclease I footprinting and transfection analyses demonstrate the presence of a negative GRE and an associated negative accessory factor element in the -271 to -225 promoter region, which inhibit the glucocorticoid response. This appears to be the first report of a promoter that contains both positive and negative GREs, which function within the same cellular environment. We hypothesize that targeted signaling to the negative accessory element within the GRU may provide tight regulation of the glucocorticoid stimulation. PMID- 16037131 TI - Transforming growth factor-beta promotes inactivation of extracellular thyroid hormones via transcriptional stimulation of type 3 iodothyronine deiodinase. AB - Thyroid hormone is a critical mediator of cellular metabolism and differentiation. Precise tissue-specific regulation of the concentration of the active ligand, T(3), is achieved by iodothyronine monodeiodination. Type 3 iodothyronine deiodinase (D3) is the major inactivating pathway, preventing activation of the prohormone T(4) and terminating the action of T(3). Using nontransformed human cells, we show that TGF-beta stimulates transcription of the hDio3 gene via a Smad-dependent pathway. Combinations of Smad2 or Smad3 with Smad4 stimulate hDio3 gene transcription only in cells that express endogenous D3 activity, indicating that Smads are necessary but not sufficient for D3 induction. TGF-beta induces endogenous D3 in diverse human cell types, including fetal and adult fibroblasts from several tissues, hemangioma cells, fetal epithelia, and skeletal muscle myoblasts. Maximum stimulation of D3 by TGF-beta also requires MAPK and is synergistic with phorbol ester and several mitogens known to signal through transmembrane receptor tyrosine kinases but not with estradiol. These data reveal a previously unrecognized interaction between two pluripotent systems, TGF-beta and thyroid hormone, both of which have major roles in the regulation of cell growth and differentiation. PMID- 16037132 TI - CHIP (carboxyl terminus of Hsc70-interacting protein) promotes basal and geldanamycin-induced degradation of estrogen receptor-alpha. AB - In estrogen target cells, estrogen receptor-alpha (ERalpha) protein levels are strictly regulated. Although receptor turnover is a continuous process, dynamic fluctuations in receptor levels, mediated primarily by the ubiquitin-proteasome pathway, occur in response to changing cellular conditions. In the absence of ligand, ERalpha is sequestered within a stable chaperone protein complex consisting of heat shock protein 90 (Hsp90) and cochaperones. However, the molecular mechanism(s) regulating ERalpha stability and turnover remain undefined. One potential mechanism involves CHIP, the carboxyl terminus of Hsc70 interacting protein, previously shown to target Hsp90-interacting proteins for ubiquitination and proteasomal degradation. In the present study, a role for CHIP in ERalpha protein degradation was investigated. In ER-negative HeLa cells transfected with ERalpha and CHIP, ERalpha proteasomal degradation increased, whereas ERalpha-mediated gene transcription decreased. In contrast, CHIP depletion by small interference RNA resulted in increased ERalpha accumulation and reporter gene transactivation. Transfection of mutant CHIP constructs demonstrated that both the U-box (containing ubiquitin ligase activity) and the tetratricopeptide repeat (TPR, essential for chaperone binding) domains within CHIP are required for CHIP-mediated ERalpha down-regulation. In addition, coimmunoprecipitation assays demonstrated that ERalpha and CHIP associate through the CHIP TPR domain. In ERalpha-positive breast cancer MCF7 cells, CHIP overexpression resulted in decreased levels of endogenous ERalpha protein and attenuation of ERalpha-mediated gene expression. Furthermore, the ERalpha-CHIP interaction was stimulated by the Hsp90 inhibitor geldanamycin (GA), resulting in enhanced ERalpha degradation; this GA effect was further augmented by CHIP overexpression but was abolished by CHIP depletion. Finally, ERalpha dissociation from CHIP by various ERalpha ligands, including 17beta-estradiol, 4 hydroxytamoxifen, and ICI 182,780, interrupted CHIP-mediated ERalpha degradation. These results demonstrate a role for CHIP in both basal and GA-induced ERalpha degradation. Furthermore, based on our observations that CHIP promotes ERalpha degradation and attenuates receptor-mediated gene transcription, we suggest that CHIP, by modulating ERalpha stability, contributes to the regulation of functional receptor levels, and thus hormone responsiveness, in estrogen target cells. PMID- 16037133 TI - Monitoring activated clotting time for combined heparin and aprotinin application: an in vitro evaluation of a new aprotinin-insensitive test using SONOCLOT. AB - The kaolin-based activated clotting time (ACT) is commonly used for monitoring heparin-induced anticoagulation alone and combined with aprotinin during cardiopulmonary bypass. However, aprotinin prolongs ACT measurements. Recently, a new so-called 'aprotinin-insensitive' ACT test (SaiACT) has been developed for the SONOCLOT analyzer. In this study we evaluated and compared this new test for the SONOCLOT analyzer in vitro with an established kaolin-based ACT from HEMOCHRON (HkACT). Twenty-five patients undergoing elective valve surgery donated 80 mL of blood after induction of anesthesia. The blood was withdrawn in citrated tubes and processed to analyze effects of heparin (0, 1, 2, and 3 U x mL(-1)), aprotinin (0, 200 kIU x mL(-1)), and 25% hemodilution with calcium-free lactated Ringer's solution on ACT measurements. A total of 400 blood samples were analyzed and ACT was measured in a wide, clinically relevant range in duplicate with SaiACT and HkACT. Addition of aprotinin to heparinized blood samples induced no significant changes of SaiACT measurements. By contrast, HkACT readings increased significantly: aprotinin prolonged HkACT in heparinized blood samples by 20% +/- 37% (2 U x mL(-1)) and 24% +/- 18% (3 U x mL(-1)), respectively, and in vitro hemodilution increased this effect. IMPLICATIONS: Current standard techniques to measure heparin-induced anticoagulation during cardiopulmonary bypass are affected by aprotinin, a drug widely used in this setting. The aim of this study was to investigate in vitro a new, so-called 'aprotinin-insensitive' test from SONOCLOT to measure heparin-induced anticoagulation more reliably in combination with aprotinin. PMID- 16037134 TI - The inhibitory effects of sevoflurane on angiotensin II- induced, p44/42 mitogen activated protein kinase-mediated contraction of rat aortic smooth muscle. AB - Sevoflurane dilates blood vessels and reduces arterial blood pressure in a dose dependent manner. Angiotensin II (Ang II) is one of the primary regulators of vascular tension and arterial blood pressure, and the p44/42 mitogen-activated protein kinases (p44/42 MAPK) are involved in Ang II-mediated vascular smooth muscle contraction. We designed this study to examine the effects of sevoflurane on Ang II-induced, p44/42 MAPK-mediated contraction of rat aortic smooth muscle. The effects of the p44/42 MAPK kinase (MEK1/2) inhibitor, PD 098059 (10(-5) molar [M], 5 x 10(-5) M and 10(-4) M), and sevoflurane (1.7%, 3.4%, and 5.1%) on Ang II induced contraction and p44/42 MAPK phosphorylation were tested in rat aortic smooth muscle, using isometric force measurement and Western blot analysis, respectively. Ang II induced both a transient contractile response and phosphorylation of p44/42 MAPK, which were significantly attenuated by PD 098059 (P < 0.05-0.01). Sevoflurane inhibited Ang II-induced contractile response in a dose-dependent manner (P < 0.05 and 0.01 in response to 3.4% and 5.1% sevoflurane, respectively). Sevoflurane also dose-dependently depressed Ang II elicited p44/42 MAPK phosphorylation (P < 0.01 in response to 3.4% and 5.1% sevoflurane). These results suggest that the inhibitory effect of sevoflurane on Ang II-induced vasoconstriction is, at least in part, caused by the inhibition of the p44/42 MAPK-mediated signaling pathway. IMPLICATIONS: The present study demonstrates that sevoflurane can dose-dependently inhibit both angiotensin II (Ang II)-induced contraction and p44/42 MAPK phosphorylation of rat aortic smooth muscle. These data suggest that sevoflurane-produced inhibition of Ang II-induced vasoconstriction is, at least in part, caused by depression of the p44/42 MAPK mediated signaling pathway. PMID- 16037135 TI - Brown-Sequard syndrome following removal of a cerebrospinal fluid drainage catheter after thoracic aortic surgery. AB - Neurological deficit remains a devastating complication of thoracic aortic surgery despite advances in methods to protect the spinal cord from ischemia. Various techniques have been used, including the combination of cerebrospinal fluid (CSF) drainage and distal aortic perfusion to decrease the incidence of postoperative neurological deficit. These deficits are usually bilateral and result in paraplegia. In this case report we present a patient with Type B aortic dissection and thoracoabdominal aortic aneurysm repair with insertion of a lumbar CSF drainage catheter. Postoperatively, the patient developed unilateral neurological features consistent with Brown-Sequard syndrome after removal of the CSF catheter. The lumbar cerebrospinal fluid catheter was reinserted and the CSF was drained. Medullary T6-7 signal abnormalities were seen on spinal cord magnetic resonance imaging, and we suggest that the spinal cord suffered a direct injury during catheter removal. The patient had an uneventful recovery. IMPLICATIONS: We describe a patient who developed unilateral neurologic features suggestive of Brown-Sequard syndrome following removal of a cerebrospinal fluid catheter after thoracic aortic surgery. We suggest that the spinal cord was injured during catheter removal. The catheter was reinserted and the patient had a full neurologic recovery. PMID- 16037136 TI - Recombinant human erythropoietin use in a critically ill Jehovah's witness after cardiac surgery. AB - Complex cardiac surgery often requires blood transfusion. Some patients refuse transfusion, even when it is potentially life-threatening to do so. Although recombinant human erythropoietin (rhEPO) has been used to reduce the need for blood transfusion, it has been considered ineffective in critically ill patients. The time course of hematological responses in a Jehovah's Witness patient with acute renal failure and severe cardiac disease suggests that a trial of rhEPO should be considered for salvage therapy in critically ill patients. IMPLICATIONS: The authors describe successful treatment of life-threatening anemia using recombinant human erythropoietin in a critically ill Jehovah's Witness patient after cardiac surgery. PMID- 16037137 TI - Solitary liver mass detected by transesophageal echocardiography. AB - Transesophageal echocardiography (TEE) has become a mainstay of cardiac surgery in the diagnosis of intracardiac and vascular lesions adjacent to the esophagus. In this case, we detected a liver mass during intraoperative TEE examination in a patient undergoing elective cardiac valve surgery. Preoperatively, the patient had normal liver function tests and no symptoms of hepatocellular carcinoma. This mass was diagnosed as hepatocellular carcinoma after the surgery and treated with transarterial chemoembolization. In conclusion, this case report outlines another potential application of intraoperative TEE extending its role outside the realm of cardiac surgery. IMPLICATIONS: A liver mass was detected during intraoperative transesophageal echocardiography (TEE) examination in a patient undergoing cardiac valve surgery. This incidental finding permitted the early diagnosis of hepatocellular carcinoma, and demonstrates another potential utility of TEE. PMID- 16037138 TI - The usefulness of intraoperative transesophageal echocardiography to identify the site of drainage of coronary artery fistula. PMID- 16037139 TI - Chasing the tumor thrombus. PMID- 16037140 TI - Ultrasound-guided epidural catheter insertion in children. AB - Epidural catheters (EC) are often used in pediatric patients for intraoperative and postoperative pain relief. The small anatomical structures and catheter insertion under general anesthesia make it more difficult to perform EC and to prevent damage. In this study we investigated the use of ultrasound (US) in detecting neuraxial structures during insertion and placement of EC in children. ASA I-II children scheduled for elective surgery under combined general and epidural anesthesia were studied. Patients received balanced anesthesia using sevoflurane, opioids and rocuronium. Before EC insertion US examination in a lateral position was done to visualize and identify neuraxial structures. Quality of visualization and site and depth of structures were recorded. Using a sterile kit to hold the US probe in position and enable the visualization of the neuraxial structures, an epidural cannula was inserted, using the loss of resistance technique, as the EC passed under US control to the desired level. Of 25 children, 23 were evaluated. Epidural space, ligamentum flavum, and dural structures were clearly identified and the depth to skin level estimated in all patients. Loss of resistance was visualized in all patients with a lumbar epidural approach. Correlation of US measured depth and depth of loss of resistance was 0.88. In eight of 23 patients EC could be visualized during insertion and in 11 others it could be visualized with additional US planes. US is an excellent tool to identify neuraxial structures in both infants and children. The size and the incomplete ossification of the vertebra allow exact visualization and localization of the depth of the epidural space, the loss of resistance, and all relevant neuraxial structures. IMPLICATIONS: Epidural catheters in children are mostly inserted under sedation or general anesthesia. This study showed that the use of ultrasound could help visualize all relevant neuraxial structures and their site and depth from the skin. PMID- 16037141 TI - Injury pattern of the neonatal brain after hypothermic low-flow cardiopulmonary bypass in a piglet model. AB - Low-flow cardiopulmonary bypass (LF-CPB) is a widely used modality in neonatal heart surgery. While facilitating surgical repair, it poses a risk of neurological injury caused by hypoperfusion. In the present study, we characterize the injury pattern and influencing factors in a piglet hypothermic LF-CPB model. Piglets were anesthetized, tracheally intubated, ventilated, and prepared for CPB. After LF-CPB for 150 min at 22 degrees C (brain) using pH-stat strategy, animals were allowed to survive for 2 or 9 days. Neurological status was assessed daily and magnetic resonance imaging scans were performed. Brains were assessed histologically. Functional neurological impairment was seen in 64%, 30%, and 0% of animals 1, 2, and 9 days after CPB, respectively. All animals showed histological brain damage, predominantly in neocortex and hippocampus, less so in basal ganglia, thalamus, white matter, and cerebellum. Cell death appeared as selective neuronal necrosis in the deeper layers in neocortex and CA1 4 sections in hippocampus. Even in a pH-stat strategy, less neocortical and hippocampal damage correlated with higher arterial partial pressure for carbon dioxide. Less hippocampal damage was associated with higher blood glucose levels. Less functional neurological impairment and basal ganglia damage correlated with higher postoperative hematocrit. IMPLICATIONS: Neuronal injury after hypothermic low-flow cardiopulmonary bypass in a piglet model using pH-stat strategy occurs predominantly in deep neocortex and hippocampus. Factors mitigating injury were higher arterial carbon dioxide, hematocrit, and blood glucose levels. PMID- 16037142 TI - Sevoflurane preconditioning limits intracellular/mitochondrial Ca2+ in ischemic newborn myocardium. AB - Sevoflurane preconditioning (SPC) in adult hearts reduces myocardial ischemia/reperfusion (I/R) injury, an effect that may be mediated by reductions in intracellular Ca(2+) ([Ca(2+)](i)) and/or mitochondrial Ca(2+) ([Ca(2+)](m)) accumulation during ischemia and reperfusion. Because the physiology, pharmacology, and metabolic responses of the newborn differ from adults, we tested the hypothesis that SPC protects newborn myocardium by limiting [Ca(2+)](i) and [Ca(2+)](m) by a K(ATP) channel-dependent mechanism. Fluorescence spectrofluorometry and nuclear magnetic resonance spectroscopy were used to measure [Ca(2+)](i), [Ca(2+)](m), and adenosine triphosphate (ATP) in 4- to 7-day old Langendorff-perfused rabbit hearts. Three experimental groups were used to study the effect of SPC on [Ca(2+)](m)/[Ca(2+)](i), ATP, as well as hemodynamics and ischemic injury. The role of mitochondrial K(ATP) channels was assessed by exposing the SPC hearts to the mitochondrial K(ATP) channel blocker 5 hydroxydecanoic acid. Our results show that SPC significantly decreased [Ca(2+)](i) and [Ca(2+)](m) during I/R, as well as decreased creatine kinase release during reperfusion and resulted in higher ATP. 5-Hydroxydecanoic acid abolished the effect of SPC on [Ca(2+)], hemodynamics, ATP, and creatine kinase release. In conclusion, decreased [Ca(2+)](i) and [Ca(2+)](m) observed with SPC is associated with greater ATP recovery as well as diminished cell injury. Mitochondrial K(ATP) channel blockade attenuates the SPC effect during I/R, suggesting that these channels are involved in the protective effects of SPC in the newborn. IMPLICATIONS: The results of this study support the hypothesis that sevoflurane preconditioning protects newborn hearts from calcium overload and ischemic injury via a mechanism dependent on mitochondrial KATP channels. PMID- 16037143 TI - A double-blind comparison of intravenous ondansetron and placebo for preventing postoperative emesis in 1- to 24-month-old pediatric patients after surgery under general anesthesia. AB - We assessed the efficacy and safety of ondansetron (0.1 mg/kg IV) prophylactically administered before surgery for prevention of postoperative vomiting (POV) in a double-blind, placebo-controlled study of 670 pediatric patients, 1- to 24-mo-old, undergoing elective surgery under general anesthesia. The study enrolled 335 children in each treatment group (ondansetron versus placebo). Significantly fewer children treated with ondansetron exhibited emesis or discontinued the study prematurely after surgery (ondansetron, 11%; placebo, 28%; odds ratio = 0.33; P < 0.0001). The number required to treat prophylactically with ondansetron to prevent POV was approximately six. Ondansetron treatment also resulted in fewer patients requiring rescue medication or assumed to have had rescue upon early discontinuation from the study during the postoperative period (ondansetron, 5%; placebo, 10%) and less emesis (0 of 6) after rescue medication when compared with placebo (7 of 21). The incidence of POV and other antiemetic effects of ondansetron were similar in children aged 1 12 mo and 13-24 mo and in children prospectively expected or not expected to require opioids as part of their anesthetic or analgesic management. Ondansetron was well tolerated; the incidence of adverse events considered possibly related to study drug was similar between treatment groups (ondansetron, 1.8%; placebo, 1.5%). IMPLICATIONS: This prospective, randomized, double-blind, placebo controlled study establishes the efficacy and tolerability of IV ondansetron (0.1 mg/kg) in the prevention of postoperative emesis in 1- to 24-mo-old pediatric patients undergoing elective surgery under general anesthesia. PMID- 16037144 TI - Single-lung ventilation for pulmonary lobe resection in a newborn. AB - The increasing frequency of video-assisted thoracoscopic interventions as well as open thoracic surgical procedures in children demands appropriate anesthetic techniques to provide single-lung ventilation. A fiberoptically directed, wire guided 5F endobronchial blocker for use in small infants has recently been devised. We report on the very special aspects of airway management in a newborn 3000-g infant who presented a major anesthetic and surgical challenge because of congenital emphysema of the left upper pulmonary lobe. IMPLICATIONS: The special aspects of single-lung ventilation in a newborn 3000-g infant who presented a major anesthetic and surgical challenge because of congenital emphysema of the left upper pulmonary lobe are reported. PMID- 16037145 TI - Remifentanil as a single drug for extracorporeal shock wave lithotripsy: a comparison of infusion doses in terms of analgesic potency and side effects. AB - This randomized, double-blind study was designed to evaluate analgesic effectiveness and side effects of two remifentanil infusion rates in patients undergoing extracorporeal shock wave lithotripsy (ESWL) for renal stones. We included 200 patients who were administered remifentanil either 0.05 microg x kg( 1) x min(-1) (n = 100) or 0.1 microg x kg(-1) x min(-1) (n = 100) plus demand bolus of 10 microg of remifentanil via a patient-controlled analgesia (PCA) device. No other sedating drugs were given. The frequencies of PCA demands and deliveries were recorded. Arterial blood pressure, oxygen saturation, and respiratory rate were recorded throughout the procedure; postoperative nausea and vomiting (PONV), dizziness, itching, agitation, and respiratory depression were measured posttreatment. Visual analog scale (VAS) scores were taken preoperatively, directly postoperatively, and 30 min after finishing the procedure. There were no statistically significant differences in the frequency of PCA demands and delivered boluses or among perioperative VAS scores. The extent of PONV and frequency of dizziness and itching immediately after and dizziness 30 min after the end of treatment were significantly reduced in the smaller dose group. We conclude that a remifentanil regimen of 0.05 microg x kg( 1) x min(-1) plus 10 microg demands is superior to 0.1 microg x kg(-1) x min(-1) plus demands, as there was no difference in the VAS scores recorded between groups and it has a less frequent incidence of side effects in patients receiving ESWL. IMPLICATIONS: Remifentanil is an appropriate analgesic choice for patients undergoing extracorporeal shock wave lithotripsy (ESWL) therapy, as it has both fast onset and offset times. We studied remifentanil as a sole drug for ESWL and have shown that an infusion rate of 0.05 microg x kg-1 x min-1 plus patient controlled analgesia demands of 10 microg provides adequate analgesia and has significantly less side effects than a dose of 0.1 microg x kg-1 x min-1 plus 10 microg demands. PMID- 16037146 TI - The effect of different isoflurane-fentanyl dose combinations on early recovery from anesthesia and postoperative adverse effects. AB - We evaluated the effect of different combinations of fentanyl-isoflurane on early recovery from anesthesia in 80 adult patients undergoing laparoscopic cholecystectomy. Anesthesia was induced with fentanyl 2 microg/kg and thiopental 5 mg/kg. Nitrous oxide was not used and patients were randomly assigned to one of four groups: Group 1 (n = 20) received 0.6% end-tidal isoflurane plus fentanyl, Group 2 (n = 20) received 1.2% end-tidal isoflurane plus fentanyl, Group 3 (n = 20) received 1.8% end-tidal isoflurane plus fentanyl, and Group 4 (n = 20) received only isoflurane. In Groups 1, 2 and 3 isoflurane concentration was kept constant and fentanyl was given as necessary to maintain the mean arterial blood pressure within +/- 10% of the minimum mean arterial blood pressure measured in the ward. In Group 4, isoflurane concentration was adjusted to maintain mean arterial blood pressure as above. At the end of skin closure isoflurane was discontinued and the time to spontaneous breathing (TSB), time to extubation (TE) and time to eye opening (TEO) were recorded. In the postanesthesia care unit, the degree of sedation, respiratory rate, Spo(2), emesis, pain, and morphine consumption were evaluated every 15 min for 1 h, and thereafter every 30 min until discharge. Fentanyl requirements were 8.3 +/- 4.5 microg/kg (mean +/- sd) in Group 1, 3.8 +/- 1.3 microg/kg in Group 2, and 3.0 +/- 0.7 microg/kg in Group 3 (P < 0.001), whereas in Group 4 the mean end-tidal concentration of isoflurane was 2.0% +/- 0.4%. Although the mean TSB was <5.5 min in all groups, TE increased from 7.3 +/- 5.1 min in Group 1 to 20.6 +/- 10.7 min in Group 4 (P < 0.001), and TEO increased from 7.4 +/- 5.1 min in Group 1 to 25.8 +/- 9.4 min in Group 4 (P < 0.001). There were no differences among the groups in any of the variables measured in the postanesthesia care unit. This study shows that the combination of a small concentration of isoflurane and a relatively larger dose of fentanyl results in a faster recovery from anesthesia than the inverse combination of doses. IMPLICATIONS: A fast recovery from anesthesia increases patient safety. This study shows that the combination of a small concentration of isoflurane and a relatively larger dose of fentanyl results in a faster recovery from anesthesia than the inverse combination of doses. PMID- 16037147 TI - The influence of gender on loss of consciousness with sevoflurane or propofol. AB - Studies have suggested that hypnotic requirements for general anesthesia and emergence may be influenced by gender. In this study, we examined the effect of gender on the hypnotic requirement for loss of consciousness (LOC) using either a volatile (sevoflurane) or an IV (propofol) anesthetic. One-hundred-fifteen unpremedicated, ASA physical status I-II patients, aged 18-40 yr old, received either sevoflurane by mask to a predetermined end-tidal concentration (%ET(sevo)) or propofol by target-controlled infusion (effect site) while breathing spontaneously. After sufficient time for equilibration, LOC was assessed by lack of response to mild prodding. The up-down method of Dixon was used to determine the hypnotic target concentration at 50% response (LOC(50)). No statistically significant difference in LOC(50) was noted between men and women for sevoflurane (0.83% +/- 0.1% and 0.92% +/- 0.09% ET, respectively). Men required significantly more propofol than women (2.9 +/- 0.2 versus 2.7 +/- 0.1 microg/mL, respectively). However, there was no difference in the bispectral index (BIS) at LOC for men or women with either hypnotic anesthetic. This investigation identified a small, statistically significant difference in hypnotic requirement at LOC(50) between men and women with propofol but not with sevoflurane. As defined by BIS, men and women had equivalent hypnotic states at LOC(50), indicating that gender had no clinically significant effect on hypnotic requirements. However, BIS at a defined clinical end-point (LOC(50)) was significantly different between the sevoflurane and propofol groups, suggesting that neurophysiological effects of these anesthetics may be different. IMPLICATIONS: Gender affects the dosing requirements for, and response to, many drugs used in anesthetic practice. Loss of consciousness is an early clinical marker of hypnotic drug effect. We found no significant difference to either an inhaled (sevoflurane) or IV (propofol) anesthetic related to patient gender. PMID- 16037148 TI - Does yawning represent a transient arousal-shift during intravenous induction of general anesthesia? AB - Although yawning occurs frequently during the IV induction of general anesthesia, the significance of this response remains unknown. In this study, we induced 30 surgical patients with 4 mg/kg thiopental IV, and 30 patients with 2 mg/kg propofol IV. Thereafter, the occurrence of yawning was continuously assessed, as the only clinical end-point, for 1 min. The electroencephalographic bispectral index was monitored throughout the observation period. The criterion for an arousal response was a transient increase during a continuing decrease in the bispectral index value. On the basis of this criterion, the sensitivity and specificity of the yawning response as an arousal sign were 77% and 80%, respectively. If a patient exhibited a yawning response, the chance of arousal was 84% (positive predictive value). With no yawning response, the chance of nonarousal was 71% (negative predictive value). According to simple logistic regression, the yawning response was predictive of a transient arousal-shift with an odds ratio of 13.5 (95% confidence interval: 3.8-48; P < 0.001). The occurrence of a yawning response during IV induction may be a clinical indicator of a transient arousal-shift during progressive loss of consciousness. IMPLICATIONS: Yawning elicited by IV anesthetic induction was related to a transient increase during the continuing decrease in the electroencephalographic bispectral index value (sensitivity and specificity, 77% and 80%, respectively). This type of yawning may be a clinical indicator of a transient arousal-shift during progressive loss of consciousness. PMID- 16037149 TI - Changes in concentrations of free propofol by modification of the solution. AB - Because free propofol is thought to be responsible for pain on injection, we investigated the changes in concentrations of free propofol by modifying two kinds of propofol products in a medium- and long-chain triglyceride (MCT/LCT) emulsion and in an LCT emulsion. The techniques used in this study were 1) mixing 2% lidocaine (10:1), 2) mixing 5% dextrose in acetated Ringer's solution to reduce pH (10:1), and 3) changing the temperature to 4 degrees , 20 degrees , and 36 degrees C. The propofol preparations were dialyzed for 24 h, and the receptor medium was analyzed using high-performance liquid chromatography. The concentration of free propofol in propofol MCT/LCT was significantly smaller by 30% than that in propofol LCT. Neither mixing lidocaine nor cooling reduced the concentrations of free propofol in both products, but the concentrations were reduced by a decrease in pH and by an increase in temperature. Because mixing lidocaine can induce instability in an emulsion of propofol and warming can rapidly induce microbial growth, injection of lidocaine before propofol administration is recommended to reduce the pain on injection. The concentrations of free propofol in propofol MCT/LCT were significantly smaller (by approximately 30%-45%) than those in propofol LCT during any situation in this study. IMPLICATIONS: Neither mixing lidocaine nor cooling reduced the concentrations of free propofol in both products but the concentrations were reduced by a decrease in pH and by an increase in temperature. Propofol medium- and long-chain triglycerides had significantly smaller concentrations by approximately 30%-45% than those in propofol long-chain triglycerides during any situation in this study. PMID- 16037150 TI - Preoperative "fentanyl challenge" as a tool to estimate postoperative opioid dosing in chronic opioid-consuming patients. AB - When opioids are used for postoperative pain control, it is useful to define the dose-response relationship for analgesia and respiratory depression. We studied 20 chronically opioid-consuming patients having elective multilevel spine fusion. Preoperatively, each patient received a fentanyl infusion of 2 microg x kg(-1) x min(-1) until the respiratory rate was <5 breaths/min. Pharmacokinetic simulations were used to estimate the effect site concentration at the time of respiratory depression and to predict the patient-controlled analgesia settings that would provide an effect-site fentanyl concentration that was 30% of the concentration associated with respiratory depression. Postoperatively, patient controlled analgesia settings were adjusted to achieve 2-3 demand doses per hour. At steady-state patient-controlled analgesia settings, arterial blood gases and plasma fentanyl levels were measured. Sixteen patients required no adjustment or one patient-controlled analgesia adjustment. The median arterial Pco(2) level was 41 mm Hg and the interquartile range was 39-46 mm Hg. Plasma fentanyl levels demonstrated a significant correlation to the estimated effect-site concentration associated with respiratory depression determined during the preoperative fentanyl challenge. A preoperative fentanyl challenge used with pharmacokinetic simulations may be a useful tool to individualize the administration of analgesics to chronically opioid-consuming patients. IMPLICATIONS: In chronically opioid-consuming patients, doses causing respiratory depression and analgesia may differ from those in opioid-naive individuals. A preoperative infusion of fentanyl, used in conjunction with pharmacokinetic simulation, may be a valuable tool for identifying clinical end-points, such as respiratory depression and analgesia, and individualizing postoperative treatment of pain in patients who chronically consume opioids. PMID- 16037151 TI - A rapid increase in the inspired concentration of desflurane is not associated with epileptiform encephalogram. AB - The large inspired concentration of sevoflurane (S) during mask induction of anesthesia can induce epileptiform electroencephalogram (EEG) associated with tachycardia. Tachycardia is also seen when the concentration of desflurane (D) is abruptly increased. It is not known whether this is associated with epileptiform EEG similar to S. We studied EEG and heart rate (HR) during rapidly increased concentrations of S or D in 31 females during the postintubation period of anesthesia. Anesthesia was induced with propofol and remifentanil, and the tracheas were intubated. Patients were randomized to receive either S or D in nitrous oxide-oxygen mixture after intubation, at a small dose first. After 10 min, S or D vaporizer was advanced to the highest reading of the vaporizer (7% for S, 18% for D) for 5 min. HR and EEG were recorded. Epileptiform EEG activity was recorded in eight of 15 patients in group S and in none in group D (P < 0.05). HR increased in both groups. In group S, HR increased gradually and the highest HR value was 84 bpm at 5 min after the increase in sevoflurane concentration. In group D, HR increased to 93 bpm 2 min after the increase in desflurane concentration (no significant difference, S versus D). A rapid increase in the concentration of S frequently induces epileptiform EEG during normoventilation. Tachycardia during increasing concentrations of D is not associated with epileptiform EEG. IMPLICATIONS: A rapid increase in the concentration of sevoflurane induces epileptiform encephalogram (EEG) with tachycardia. A rapid increase in the concentration of desflurane also induces tachycardia but is not associated with epileptiform EEG. PMID- 16037152 TI - The gamma-subunit governs the susceptibility of recombinant gamma-aminobutyric acid type A receptors to block by the nonimmobilizer 1,2 dichlorohexafluorocyclobutane (F6, 2N). AB - To identify anesthetic effects that produce the different components of the complex anesthetic state, the so-called nonanesthetics/nonimmobilizer classes of compounds have been introduced. Because ionotropic gamma-aminobutyric acid type A (GABA(A)) receptors play an important role in the mediation of the central nervous system (CNS) effects of general anesthetics, and their susceptibility to modulation by various drugs depends on subunit composition, we have compared the effect of the nonimmobilizer 1,2-dichlorohexafluorocyclobutane (F6) on GABA(A) receptors expressed in human embryonic kidney 293 cells transfected with alpha1beta2 versus alpha1beta2gamma2s subunits. Using rapid perfusion and whole cell recording techniques, we found that, like isoflurane, F6 blocked GABA induced currents through alpha1beta2 receptors but, unlike isoflurane, the presence of the gamma2s subunit conferred complete resistance to block by F6. Also, in contrast to isoflurane, F6 had no effect on deactivation kinetics of GABA-induced currents in either type of receptor. We conclude that modulation of alphabetagamma receptors plays little or no role in the actions of F6, but the block of alphabeta receptors may contribute to its effects on the CNS. IMPLICATIONS: Gamma-aminobutyric acidA receptors are the target of numerous drugs affecting the central nervous system. The subunit composition of the GABAA receptors governs their interaction with many drugs. We investigated whether the gamma-subunit influences the interaction with the nonimmobilizer F6. PMID- 16037153 TI - The role of nicotinic inhibition in ketamine-induced behavior. AB - Several anesthetic drugs are nicotinic antagonists at or below levels used for anesthesia, including ketamine and volatile anesthetics. In contrast, propofol does not inhibit nicotinic receptors. To determine the potential behavioral ramifications of nicotinic inhibition by ketamine, we determined the doses of ketamine required to induce immobility, impair the righting reflex, and cause analgesia in the absence and presence of several nicotinic ligands. Propofol was used as a control in similar experiments. When used as a sole anesthetic drug, 383 +/- 22 mg/kg ketamine intraperitoneally (IP) was required for immobility and 180 +/- 17 mg/kg IP impaired righting reflex. Propofol, 371 +/- 34 mg/kg IP, induced immobility whereas 199 mg/kg IP inhibited the righting reflex. Nicotinic antagonists had no effect on the dose of propofol or ketamine required for either end-point. When nociceptive responses were tested at subhypnotic doses, no pronociceptive or antinociceptive phase was identified for propofol, whereas analgesia was induced at ketamine doses larger than 60 mg/kg IP. The broad spectrum nicotinic antagonist mecamylamine enhanced the analgesic action of ketamine. These findings are different than those seen with volatile anesthetics, where nicotinic inhibition is thought to be responsible for a pronociceptive action. Such a phase is possibly obscured by analgesia induced as a result of N methyl-d-aspartic acid antagonism by ketamine. IMPLICATIONS: Ketamine and volatile anesthetics, but not propofol, inhibit neuronal nicotinic acetylcholine receptors in clinically relevant concentration ranges. Nicotinic inhibition by ketamine is not related to its immobilizing or sedating effects but may play a role in ketamine's analgesic action. PMID- 16037154 TI - Beta3-containing gamma-aminobutyric acidA receptors are not major targets for the amnesic and immobilizing actions of isoflurane. AB - Mice bearing an N265M point mutation in the gamma-aminobutyric acid (GABA)(A) receptor beta3 subunit resist various anesthetic effects of propofol and etomidate. They also require a 16% larger concentration of enflurane and a 21% larger concentration of halothane to abolish the withdrawal reflex than do wild type mice. Using a Pavlovian test, we measured whether this mutation increased the concentration of isoflurane required to impair learning and memory relative to wild-type mice. We found that the concentration was not significantly increased. We also measured MAC (the minimum alveolar concentration required to eliminate movement in response to noxious stimulation in 50% of subjects). Isoflurane MAC for mutant mice (1.93% +/- 0.0.03%; mean +/- se; n = 14) was 17.0% larger than MAC for wild-type mice (1.65 +/- 0.04; n = 14; P < 0.001). Similarly, the cyclopropane MAC for mutant mice (27.6% +/- 0.55%; n = 16) was 13.6% larger than MAC for wild-type mice (24.3 +/- 0.46; n = 8; P < 0.01). The increase in MAC for cyclopropane was unexpected, because published reports find only minimal actions at alpha1beta2gamma2 GABA(A) receptors whereas isoflurane provides a large enhancement. Consistent with previous work on alpha1beta2gamma2 GABA(A) receptors, we found in Xenopus oocytes that 5 MAC cyclopropane enhanced the effect of GABA on alpha1beta2gamma2 GABA(A) receptors by only 76%, and by a nearly identical enhancement in alpha1beta3gamma2, and alpha6beta3gamma2 receptors. In contrast, a much smaller concentration of isoflurane (1 MAC) produced a 160% to 310% enhancement in these receptors. If, relative to isoflurane, cyclopropane minimally increases GABA-induced chloride currents at any GABA(A) receptor subtype, the present data for MAC are consistent with the notion that GABA(A) receptors do not mediate the immobility produced by inhaled anesthetics. IMPLICATIONS: The results of the present study indicate that beta3 containing gamma-aminobutyric acidA receptors do not mediate the amnesia produced by isoflurane and do not mediate, or only partially mediate, the immobility produced by inhaled anesthetics. PMID- 16037155 TI - The Influence of desflurane on QTc interval. AB - Volatile anesthetics may prolong the QTc interval and this may result in grave cardiac arrhythmias. We assessed the effect of desflurane on the QTc interval in 40 ASA physical status I or II patients. Volatile anesthetic induction with desflurane was performed, and after obtaining adequate level of anesthesia, QTc interval, heart rate, and noninvasive arterial blood pressure were measured. Prolongation of the QTc interval was observed within the first minute of anesthesia. There were no differences in QTc interval changes between sexes at any time. We conclude that desflurane prolongs the QTc interval, but that there are no differences between genders in sensitivity to this action. IMPLICATIONS: We assessed the effect of desflurane on QTc interval in patients without cardiac diseases. Prolongation of the interval was evident by the first minute of desflurane anesthesia. There were no differences between female and male patients. PMID- 16037156 TI - A prospective, randomized, double-blind, placebo-controlled study of the platelet and global hemostatic effects of Ganoderma lucidum (Ling-Zhi) in healthy volunteers. AB - Ganoderma lucidum is a Chinese herbal medicine popular with cancer patients. Previous in vitro studies suggested that Ganoderma lucidum might impair hemostasis. In this prospective, randomized double-blind study, healthy volunteers received orally Ganoderma lucidum capsules 1.5 g (n = 20) or placebo (n = 20) daily for 4 wk. We monitored subjects before drug administration and at 4 and 8 wk thereafter by routine coagulation screen, fibrinogen concentration, von Willebrand ristocetin cofactor activity, platelet function analyzer PFA-100, and thrombelastography. There were no significant between-group differences and all measurements remained within the normal range. Ganoderma lucidum ingestion over 4 wk was not associated with impairment of hemostasis. IMPLICATIONS: Ingestion of Ganoderma lucidum does not cause impairment of hemostatic function in healthy volunteers, despite earlier in vitro reports that it may cause platelet inhibition and may have other antithrombotic and fibrinolytic activity. The use of Ganoderma lucidum preoperatively is unlikely to increase the risk of surgical bleeding in otherwise healthy patients. PMID- 16037157 TI - Narcotrend or bispectral index monitoring during desflurane-remifentanil anesthesia: a comparison with a standard practice protocol. AB - Bispectral Index (BIS) (Aspect Medical Systems, Newton, MA) and Narcotrend (MonitorTechnik, Bad Bramstedt, Germany) are monitoring devices that were, as others, designed to assess the depth of anesthesia. Meanwhile, a number of studies indicate that with total IV anesthesia, BIS and Narcotrend have comparable effects on drug consumption and recovery times whereas comparative clinical data for volatile anesthetics are still missing. Therefore, we designed the present prospective, randomized, and double-blinded study to compare the effects of BIS and Narcotrend monitoring during desflurane-remifentanil anesthesia and versus a standard anesthetic practice protocol. One-hundred-twenty adult patients scheduled for minor orthopedic surgery were randomized to receive a desflurane-remifentanil anesthetic controlled either by Narcotrend or by BIS or solely by clinical variables. Anesthesia was induced with 0.4 microg x kg(-1) x min(-1) remifentanil and 2 mg/kg propofol. After tracheal intubation, remifentanil was infused at a constant rate of 0.2 microg x kg(-1) x min(-1) whereas desflurane in 1.5 L/min O(2)/air was adjusted according to clinical variables or the following target values: during maintenance of anesthesia to a value of "D(0)" (Narcotrend) or "50" (BIS), 15 min before the end of surgery to "C(1)" (Narcotrend) or "60" (BIS), whereas in the standard protocol group, desflurane was controlled according to clinical variables, e.g., heart rate, arterial blood pressure, movements. Recovery times and desflurane consumption were recorded by a blinded investigator. The desflurane vaporizer was weighed before and after anesthesia and consumption per minute was calculated. Data are mean +/- sd. The groups were comparable for demographic data, duration of anesthesia, and mean remifentanil dosages. Compared with standard practice, patients with Narcotrend or BIS monitoring needed significantly less desflurane (standard practice 443 +/- 71 mg/min, Narcotrend 374 +/- 124 mg/min, BIS monitoring 416 +/- 99 mg/min desflurane [both P < 0.05]). However, recovery times were not significantly different between the groups, e.g., opening of eyes 4.7 +/ 2.2 versus 3.7 +/- 2.0 versus 4.2 +/- 2.1 min. During desflurane-remifentanil anesthesia, Narcotrend and BIS monitoring seem to be equally effective compared with standard anesthetic practice: BIS and Narcotrend allow for a small reduction of desflurane consumption whereas recovery times are only slightly reduced. IMPLICATIONS: Monitoring the electroencephalogram with Narcotrend or Bispectral Index during desflurane-remifentanil anesthesia only slightly reduces recovery times when compared with a standard practice protocol. PMID- 16037158 TI - The impact of acoustic stimulation on the AEP monitor/2 derived composite auditory evoked potential index under awake and anesthetized conditions. AB - The AEP Monitor/2 features an auditory evoked potential (AEP) and electroencephalogram (EEG)-derived hybrid index of the patient's hypnotic state. The composite AEP index (AAI) is preferably calculated from the AEP, but in case of low signal quality it is based entirely on the spontaneous EEG. We investigated the impact of auditory input on the AAI in 16 patients with correctly positioned headphones for acoustic stimulation and headphones disconnected from the patient's ears under awake and anesthetized conditions. The AAI and the Narcotrend Index (NI), another EEG-based measure of hypnotic depth, were recorded simultaneously. AAI values under awake and anesthetized conditions were higher with correctly positioned headphones than with headphones disconnected from the patient's ears (P < 0.05) but remained within the range indicating the patient's actual hypnotic state as given by the manufacturer of the monitor. Under awake conditions with correctly positioned headphones we observed frequent fluctuations between AEP-derived and EEG-derived AAI, whereas with headphones disconnected from the patient's ears the AAI calculation was completely EEG based. Acoustic stimulation had no impact on the Narcotrend Index. Although relevant misinterpretations of the patient's hypnotic state as a consequence of a turnover from AEP-derived to EEG-derived AAI values should not occur, an improved harmonization of the two methods of indexing would be desirable. IMPLICATIONS: The AEP Monitor/2 generates an Index (AAITM) indicating the patient's hypnotic state by analyzing either auditory evoked potentials (AEP) or spontaneous electroencephalographic (EEG) activity. We demonstrate that, though significantly different under AEP-derived or EEG-derived conditions, AAI values remain within the range indicating the patient's actual hypnotic state as given by the manufacturer of the device. PMID- 16037159 TI - A pilot study of continuous transtracheal mixed venous oxygen saturation monitoring. AB - In this study, we investigated the feasibility and the accuracy of transtracheal mixed venous oxygen saturation (Svo(2)) monitoring. Ten patients undergoing thoracic surgery were included in this study. A single-use pediatric pulse oximetry sensor was attached to the double-lumen tube between the tracheal and bronchial cuff. After anesthesia was induced, the double-lumen tube was inserted into the trachea and adjusted to the proper position. During surgery, the pulmonary arterial blood was sampled every 3 min for 15 min to measure the Svo(2). The measurements made by the transtracheal pulmonary pulse oximeter (Sto(2)) were recorded at the same time that blood was sampled from the pulmonary artery for Svo(2) measurements. The levels of measurement agreement between the Sto(2) and the Svo(2) were analyzed using the Bland and Altman method. The mean +/- sd (range) oxygen saturation values during the data collecting period were 82.0% +/- 4.9% (72%-91%) for the Sto(2) and 82.2% +/- 5.5% (71%-91%) for the Svo(2), respectively. The linear correlation coefficient of the regression analysis between the Sto(2) and the Svo(2) was 0.934 (P < 0.05). A 95% confidence interval for absolute difference between the Sto(2) and the Svo(2) was 1.58% 2.09%. The mean +/- 2 sd difference between the Sto(2) and the Svo(2) was 0.12% +/- 3.97% on the Bland and Altman graph. We conclude that it is feasible to monitor the pulmonary artery oxygen saturation continuously by a transtracheal pulse oximetry technique and that it can be done so accurately. IMPLICATIONS: Mixed venous oxygen saturation (Svo2) is a measure of the balance between oxygen supply and consumption throughout the whole body. Svo2 can be measured invasively by inserting a pulmonary artery catheter with the associated disadvantages of cost and potential for patient injury. In this study, we investigated the feasibility of noninvasive Svo2 measurement using a transtracheal pulse oximetry technique. PMID- 16037160 TI - Dietary omega-3 fatty acids may be associated with increased neuropathic pain in nerve-injured rats. AB - Certain dietary proteins and oils are capable of decreasing chronic neuropathic pain levels in rats after partial sciatic nerve ligation injury. We tested, for the first time, the role of dietary polyunsaturated fatty acids in suppressing pain in partial sciatic nerve ligation-injured rats. Six groups of male Wistar rats were fed an identical casein-based, fat-free diet for 1 wk preceding partial sciatic nerve ligation injury and for 1 wk thereafter. In addition, rats received, via gavage, 1 mL/day of pure canola, corn, hemp, soy, or sunflower oil, differing significantly in their omega-3 and omega-6 polyunsaturated fatty acid content, or 1 mL of plain water. Responses to tactile and noxious heat stimuli were recorded before and after surgery and a difference score was calculated for each group by subtracting the preoperative from the post-partial sciatic nerve ligation values. Heat hyperalgesia, but not tactile allodynia, was significantly different among the dietary groups (P = 0.005). Heat hyperalgesia of rats fed hemp oil, developing the most robust response, was significantly larger compared with rats fed corn oil, developing the least pain model (difference score: 24.3 +/- 4.1 s versus 6.1 +/- 3.1 s, respectively; P < 0.001). These oils contain similar levels of omega-6 polyunsaturated fatty acids (hemp, 60%; corn, 58%) but their omega-3 levels are 28-fold different (20% versus 0.7%, respectively). A significant correlation was found among dietary levels of omega-3, but not omega 6 or the omega-3/omega-6 ratio, of the six dietary groups and heat hyperalgesia (P = 0.006). We conclude that dietary oil might predict levels of neuropathic pain in rats and that this effect may be associated with dietary omega-3 levels. IMPLICATIONS: We found that certain commonly used oils can have a significant analgesic effect in rats with persistent pain after partial nerve injury. This effect may be associated with the amounts of omega-3 fatty acids consumed by rats. PMID- 16037161 TI - The cross-modal interaction between pain-related and saccade-related cerebral activation: a preliminary study by event-related functional magnetic resonance imaging. AB - Pain-related cerebral activation in functional magnetic resonance imaging shows less consistent signals that decay earlier than in conventional task-related activation. This may result from pain's top-down inhibition mediated by cognitive or hemodynamic interaction that could affect activation by other modalities. Using event-related functional magnetic resonance imaging, we examined whether pain affects cerebral activation by a saccade task through such cross-modal interaction. Six right-handed volunteers underwent whole-brain echo-planar imaging on a 3.0 T magnetic resonance imaging scanner while they received thermal pain stimulus at 50 degrees C on the right forearm (P; n = 6), performed a visually guided saccade task (V; n = 6), and went through a simultaneous pain plus-saccade paradigm (PV; n = 5). Averaged functional activation maps were synthesized and signal time courses were analyzed at activation clusters. P activated the bilateral secondary somatosensory cortex (S2). V activated the posterior, supplementary, frontal eye fields, and visual areas. PV enhanced the S2 activation and activated additional pain-related areas, including the bilateral premotor area, right insula, anterior, and posterior cingulate cortices. In contrast, V-related activation was attenuated in PV. We propose that pain caused cross-modal suppression on the oculomotor activity and that an oculomotor task enhanced pain-related activation by triggering attention toward pain. IMPLICATIONS: Pain-related cerebral activation is enhanced by attention toward pain. It may involve top-down suppression over the unrelated neural networks of saccade. PMID- 16037162 TI - A behavioral and pharmacological validation of the acetone spray test in gerbils with a chronic constriction injury. AB - Cold and mechanical allodynia are important symptoms in patients with neuropathic pain. The study of cold allodynia in animals can help us to understand the underlying pathophysiological mechanisms of neuropathic pain and to validate drugs. The evaluation of cold allodynia in gerbils with a chronic constriction injury of the sciatic nerve on the cold plate is not always stable. We developed a new application method of acetone using a specific spray technique with an Eppendorf multistepper pipette. The chronic constriction injury of the sciatic nerve in gerbils resulted in a long-lasting mechanical and acetone spray-induced hyperreactivity throughout the testing period, which is clearly different from what was seen in sham-operated animals. The acetone spray test incorporates a multimodal stimulus different from direct cold stimulation. The reactivity to the acetone spray coincides in time and strength with the reactivity observed in mechanical allodynia in gerbils and with mechanical and thermal allodynia in other species. Furthermore, a pharmacological validation of the acetone spray test by different reference compounds was performed. Different compounds effective in neuropathic pain models in rodents influence the hyperreactivity to the acetone spray after acute and chronic administration. This study indicates that the multimodal acetone spray test is a valuable tool in the study of neuropathic pain in rodents. IMPLICATIONS: The acetone spray test is a multimodal and valuable tool in the evaluation of neuropathic pain behavior in gerbils. PMID- 16037163 TI - Validation of statistical methods to compare cancellation rates on the day of surgery. AB - We investigated the validity of several statistical methods to monitor the cancellation of electively scheduled cases on the day of surgery: chi(2) test, Fisher's exact test, Rao and Scott test, Student's t-test, Clopper-Pearson confidence intervals, and Chen and Tipping modification of the Clopper-Pearson confidence intervals. Discrete-event computer simulation over many years was used to represent surgical suites with an unchanging cancellation rate. Because the true cancellation rate was fixed, the accuracy of the statistical methods could be determined. Cancellations caused by medical events, rare events, cases lasting longer than scheduled, and full postanesthesia or intensive care unit beds were modeled. We found that applying Student's two-sample t-test to the transformation of the numbers of cases and canceled cases from each of six 4-wk periods was valid for most conditions. We recommend that clinicians and managers use this method in their quality monitoring reports. The other methods gave inaccurate results. For example, using chi(2) or Fisher's exact test, hospitals may erroneously determine that cancellation rates have increased when they really are unchanged. Conversely, if inappropriate statistical methods are used, administrators may claim success at reducing cancellation rates when, in fact, the problem remains unresolved, affecting patients and clinicians. IMPLICATIONS: Operating room cancellation rates can be monitored statistically by considering the number of canceled and performed cases during each 4-week period, performing a transformation of each period's cancellation rate, and then applying Student's t-test. Methods such as the Fisher's exact test and {chi}2 test should be avoided for this application because they can give erroneous results. PMID- 16037164 TI - The efficacy and safety of pain management before and after implementation of hospital-wide pain management standards: is patient safety compromised by treatment based solely on numerical pain ratings? AB - Inadequate analgesia in hospitalized patients prompted the Joint Commission on Accreditation of Healthcare Organizations in 2001 to introduce standards that require pain assessment and treatment. In response, many institutions implemented treatment guided by patient reports of pain intensity indexed with a numerical scale. Patient safety associated with treatment of pain guided by a numerical pain treatment algorithm (NPTA) has not been examined. We reviewed patient satisfaction with pain control and opioid-related adverse drug reactions before and after implementation of our NPTA. Patient satisfaction with pain management, measured on a 1-5 scale, significantly improved from 4.13 to 4.38 (P < 0.001) after implementation of an NPTA. The incidence of opioid over sedation adverse drug reactions per 100,000 inpatient hospital days increased from 11.0 pre-NPTA to 24.5 post-NPTA (P < 0.001). Of these patients, 94% had a documented decrease in their level of consciousness preceding the event. Although there was an improvement in patient satisfaction, we experienced a more than two-fold increase in the incidence of opioid over sedation adverse drug reactions in our hospital after the implementation of NPTA. Most adverse drug reactions were preceded by a documented decrease in the patient's level of consciousness, which emphasizes the importance of clinical assessment in managing pain. IMPLICATIONS: Although patient satisfaction with pain management has significantly improved since the adoption of pain management standards, adverse drug reactions have more than doubled. For the treatment of pain to be safe and effective, we must consider more than just a one-dimensional numerical assessment of pain. PMID- 16037165 TI - Helping surgical patients quit smoking: why, when, and how. AB - Millions of cigarette smokers undergo elective surgery each year. Efforts to help them quit smoking could improve immediate perioperative outcomes, such as those related to the cardiac and respiratory systems, and the healing of surgical wounds. Perhaps more importantly, the scheduling of elective surgery represents an excellent opportunity for smokers to permanently quit, with great benefit to their long-term health. Although it is difficult for smokers to quit, there are now several interventions of proven benefit that can more than double the chances of success. These include simple physician advice to quit, brief behavioral interventions that can be provided by physicians or other clinicians, and pharmacotherapy with drugs such as nicotine. Although specific strategies tailored for the surgical patient remain to be developed, there are steps that anesthesiologists can implement into their practices now that can help their patients quit smoking. IMPLICATIONS: The scheduling of elective surgery provides an excellent opportunity for cigarette smokers to quit, given the deleterious effects of smoking on perioperative outcome and the potential for surgery to serve as a "teachable moment" to modify smoking behavior. There are now proven methods to help them do so. PMID- 16037166 TI - Nitric oxide synthase inhibition in sepsis? Lessons learned from large-animal studies. AB - Nitric Oxide (NO) plays a controversial role in the pathophysiology of sepsis and septic shock. Its vasodilatory effects are well known, but it also has pro- and antiinflammatory properties, assumes crucial importance in antimicrobial host defense, may act as an oxidant as well as an antioxidant, and is said to be a "vital poison" for the immune and inflammatory network. Large amounts of NO and peroxynitrite are responsible for hypotension, vasoplegia, cellular suffocation, apoptosis, lactic acidosis, and ultimately multiorgan failure. Therefore, NO synthase (NOS) inhibitors were developed to reverse the deleterious effects of NO. Studies using these compounds have not met with uniform success however, and a trial using the nonselective NOS inhibitor N(G)-methyl-l-arginine hydrochloride was terminated prematurely because of increased mortality in the treatment arm despite improved shock resolution. Thus, the issue of NOS inhibition in sepsis remains a matter of debate. Several publications have emphasized the differences concerning clinical applicability of data obtained from unresuscitated, hypodynamic rodent models using a pretreatment approach versus resuscitated, hyperdynamic models in high-order species using posttreatment approaches. Therefore, the present review focuses on clinically relevant large-animal studies of endotoxin or living bacteria-induced, hyperdynamic models of sepsis that integrate standard day-to-day care resuscitative measures. PMID- 16037167 TI - Severe transfusion-related acute lung injury. AB - A 46-yr-old man developed severe hypoxemia, pulmonary infiltrates, and an acute decrease in his leukocyte count shortly after transfusion of fresh-frozen plasma (FFP) during recovery from cardiac surgery. Cardiogenic pulmonary edema was excluded. Granulocyte-reactive and agglutinating alloantibodies were detected in the serum of the fresh-frozen plasma donor. The cross-match with the patient's granulocytes revealed antibodies specific for HLA class I. Transfusion-related acute lung injury (TRALI) is a potentially life-threatening, under-recognized and under-reported complication of transfusion. Conservative transfusion strategies and preclusion of the implicated blood donors with granulocyte-reactive antibodies from future blood donation may prevent TRALI and could save lives. IMPLICATIONS: Transfusion-related acute lung injury (TRALI) is a potentially life threatening, probably under-recognized and under-reported complication of transfusing blood products. Conservative transfusion strategies and preclusion of the implicated blood donors with granulocyte-reactive antibodies from future blood donation may prevent TRALI and potentially save lives. PMID- 16037168 TI - A retrospective analysis of a remifentanil/propofol general anesthetic for craniotomy before awake functional brain mapping. AB - We performed this study to summarize drug dosing, physiologic responses, and anesthetic complications from an IV general anesthetic technique for patients undergoing craniotomy for awake functional brain mapping. Review of 98 procedures revealed "most rapid" IV infusion rates for remifentanil 0.05, 0.05-0.09 microg x kg(-1) x min(-1) and propofol 115, 100-150 microg x kg(-1) x min(-1). The infusions lasted for 78, 58-98 min. Intraoperative emergence from general anesthesia was 9 (6-13) min after discontinuing IV infusions to allow for brain mapping and was independent of infusion duration and duration of craniotomy before mapping. Spontaneous ventilation was generally satisfactory during drug infusion, as evidenced by Sao(2) = 95% (92%-98%) and Paco(2) = 50 (47-55) mm Hg. However, we recorded at least one 30-s epoch of apnea in 69 of 96 patients. Maximum systolic arterial blood pressure was 150 (139-175) mm Hg and minimal systolic arterial blood pressure was 100 (70-150) mm Hg during drug infusion. Three patients experienced intraoperative seizures. Two patients did not tolerate the awake state and required reinduction of general anesthesia. No patients required endotracheal intubation or discontinuation of surgery. This general anesthetic technique is effective for craniotomy with awake functional brain mapping and offers an alternative to continuous wakefulness or other IV sedation techniques. IMPLICATIONS: An IV general anesthetic technique using remifentanil and propofol is an effective method allowing for reliable emergence for intraoperative awake functional brain mapping during craniotomy. PMID- 16037169 TI - Sevoflurane impairs cerebral blood flow autoregulation in rats: reversal by nonselective nitric oxide synthase inhibition. AB - In this study, we investigated the effects of 1.0 and 2.0 minimum alveolar anesthetic concentration (MAC) sevoflurane on cerebral blood flow (CBF) autoregulation before and after nonselective inhibition of nitric oxide (NO) synthase in rats. Rats were randomly assigned as follows: Group 1 (n = 8): 1.0 MAC sevoflurane; Groups 2 and 3 (n = 8 per group): 2.0 MAC sevoflurane. Assessment of autoregulation within a mean arterial blood pressure range of 140 60 mm Hg was performed by graded hemorrhage before and after administration of l arginine methyl ester (l-NAME, 30 mg/kg IV, Groups 1 and 2) or during hypocapnia (Group 3). In 10 additional animals, brain tissue NO(2)(-) concentrations were measured at 1.0 and 2.0 MAC sevoflurane. CBF autoregulation was maintained with 1.0 MAC sevoflurane (Group 1) regardless of NO synthase status indicating that CBF autoregulation might not be related to NO availability. Sevoflurane dose dependently increased brain tissue NO(2)(-) and impaired CBF autoregulation. Administration of l-NAME (Group 2) but not hypocapnia (Group 3) restored CBF autoregulation. This suggests that sevoflurane impairs the autoregulatory capacity secondary to an increase of the perivascular NO availability and questions the importance of basal cerebrovascular tone in terms of vasodilatory capacity during hypotensive challenges. IMPLICATIONS: The present study suggests that the volatile anesthetic sevoflurane dose-dependently impairs cerebrovascular autoregulation by mechanisms secondary to increase of perivascular nitric oxide availability. PMID- 16037170 TI - The effects of sevoflurane and hyperventilation on electrocorticogram spike activity in patients with refractory epilepsy. AB - We investigated the effects of sevoflurane and hyperventilation on intraoperative electrocorticogram (ECoG) spike activity in 13 patients with intractable epilepsy. Grid electrodes were placed on the brain surface and ECoG was recorded under the following conditions: 1) 0.5 minimal alveolar anesthetic concentration (MAC) sevoflurane, 2) 1.5 MAC sevoflurane, and 3) 1.5 MAC sevoflurane with hyperventilation. The number of spikes per 5 min and the percentage of leads with spikes were assessed in each condition. In 4 patients with chronically implanted subdural electrodes, the leads with seizure onset and with spikes during the interictal periods in the awake state were compared with those during sevoflurane anesthesia at 0.5 MAC and 1.5 MAC. The number of spikes and the percentage of leads with spikes were significantly more under 1.5 MAC sevoflurane anesthesia compared with those under 0.5 MAC sevoflurane (P < 0.05). The induction of hyperventilation significantly increased the number of spikes and percentage of leads with spikes (P < 0.05). With 0.5 MAC sevoflurane, the leads with spikes were similar to those at seizure onset in the awake state, whereas with 1.5 MAC sevoflurane, spikes were similar to those occurring during interictal periods in the awake state. These results indicate that sevoflurane and hyperventilation can affect the frequency and extent of ECoG spike activity in patients with intractable epilepsy. Careful attention should be paid to the concentration of sevoflurane used and ventilatory status when intraoperative EcoG is used to localize epileptic lesions. IMPLICATIONS: Electrocorticogram can be used to define the location and extent of epileptic foci during epilepsy surgery. However, electrocorticogram can be affected by anesthetic technique. The present study found that sevoflurane concentration and hyperventilation affected the frequency and the extent of electrocorticogram spike activity in epileptic patients. PMID- 16037171 TI - Revising a dogma: ketamine for patients with neurological injury? AB - We evaluated reports of randomized clinical trials in the perioperative and intensive care setting concerning ketamine's effects on the brain in patients with, or at risk for, neurological injury. We also reviewed other studies in humans on the drug's effects on the brain, and reports that examined ketamine in experimental brain injury. In the clinical setting, level II evidence indicates that ketamine does not increase intracranial pressure when used under conditions of controlled ventilation, coadministration of a gamma-aminobutyric acid (GABA) receptor agonist, and without nitrous oxide. Ketamine may thus safely be used in neurologically impaired patients. Compared with other anesthetics or sedatives, level II and III evidence indicates that hemodynamic stimulation induced by ketamine may improve cerebral perfusion; this could make the drug a preferred choice in sedative regimes after brain injury. In the laboratory, ketamine has neuroprotective, and S(+)-ketamine additional neuroregenerative effects, even when administered after onset of a cerebral insult. However, improved outcomes were only reported in studies with brief recovery observation intervals. In developing animals, and in certain brain areas of adult rats without cerebral injury, neurotoxic effects were noted after large-dose ketamine. These were prevented by coadministration of GABA receptor agonists. IMPLICATIONS: Ketamine can be used safely in neurologically impaired patients under conditions of controlled ventilation, coadministration of a {gamma}-aminobutyric acid receptor agonist, and avoidance of nitrous oxide. Its beneficial circulatory effects and preclinical data demonstrating neuroprotection merit further animal and patient investigation. PMID- 16037172 TI - A comparison of Espocan and Tuohy needles for the combined spinal-epidural technique for labor analgesia. AB - When using the needle-through-needle combined spinal-epidural (CSE) technique for labor analgesia, failure to obtain cerebrospinal fluid (CSF), paresthesias, and intrathecal or intravascular migration of the catheter are of concern. Epidural needles with spinal needle apertures, such as the back-hole Espocan (ES) needles, are available and may reduce these risks. We describe the efficacy and adverse events associated with a modified epidural needle (ES) versus a conventional Tuohy needle for CSE. One-hundred parturients requesting labor analgesia (CSE) were randomized into 2 groups: 50-ES 18-gauge modified epidural needle with 27 gauge Pencan atraumatic spinal needle, 50-conventional 18-gauge Tuohy needle with 27-gauge Gertie Marx atraumatic spinal needle. Information on intrathecal or intravascular catheter placement, paresthesia on introduction of spinal needle, failure to obtain CSF through the spinal needle after placement of epidural needle, unintentional dural puncture, and epidural catheter function was obtained. No intrathecal catheter placement occurred in either group. Rates of intravascular catheter placement and unintentional dural puncture were similar between the groups. Significant differences were noted regarding spinal needle induced paresthesia (14% ES versus 42% Tuohy needles, P = 0.009) and failure to obtain CSF on first attempt (8% ES versus 28% Tuohy needles, P < 0.02). Use of ES needles for CSE significantly reduces paresthesia associated with the insertion of the spinal needle and is associated with more frequent successful spinal needle placement on the first attempt. IMPLICATIONS: The use of modified epidural needles with a back hole for combined spinal-epidural technique significantly reduces paresthesia associated with the insertion of the spinal needle and is associated with more frequent successful spinal needle placement on the first attempt. PMID- 16037173 TI - The comparative neurotoxicity of intrathecal lidocaine and bupivacaine in rats. AB - There is a considerable difference in the number of reports of neurologic injury in the literature between lidocaine and other local anesthetics. Few in vivo animal studies have produced convincing results showing a difference in neurotoxicity among anesthetics. We investigated whether lidocaine and bupivacaine differ with respect to sensory impairment and histologic damage when equipotent doses of the two are administered intrathecally in rats. First, to determine relative anesthetic potency, rats intrathecally received 20 muL of saline, 0.625%, 1.25%, 2.5%, or 5% lidocaine, or 0.125%, 0.25%, 0.5%, or 1.0% bupivacaine, and were examined with the tail-flick test for 90 min. The potency ratio calculated was approximately 1:4.70 (95% confidence interval, 3.65-6.07) for lidocaine/bupivacaine. In the next experiment, 45 rats intrathecally received 20 muL of saline, 2.13% bupivacaine (approximately 1.5 mg/kg), or 10% lidocaine (approximately 6.9 mg/kg), and were examined for persistent functional impairment and morphologic damage. Rats given lidocaine developed significantly more prolonged tail-flick latencies than those in other groups 4 days after injection and incurred more morphologic damage than those given saline or bupivacaine. In conclusion, although the doses of anesthetics administered were larger than those used clinically, the present results suggest that bupivacaine is less neurotoxic than lidocaine when administered intrathecally at equipotent concentrations in the rat model. IMPLICATIONS: Bupivacaine induces less severe functional impairment and morphologic damage than lidocaine when the two anesthetics are intrathecally administered at equipotent concentrations in the rat. PMID- 16037174 TI - The long term myotoxic effects of bupivacaine and ropivacaine after continuous peripheral nerve blocks. AB - Compared with bupivacaine, acute myotoxicity of ropivacaine is less severe. Thus, in this study we compared the long term myotoxic effects of both drugs in a clinically relevant setting. Femoral nerve catheters were inserted in anesthetized pigs, and either 20 mL of bupivacaine (5 mg/mL) or ropivacaine (7.5 mg/mL) was injected. Subsequently, bupivacaine (2.5 mg/mL) and ropivacaine (3.75 mg/mL) were continuously infused (8 mL/h) over 6 h. Control animals were treated with corresponding volumes of normal saline. After 7 and 28 days, respectively, muscle samples were dissected at the former injection sites, and histological patterns of muscle damage were blindly scored (0 = no damage to 3 = marked lesions/myonecrosis) and compared. No morphological tissue changes were detected in control animals. In the observed period, both local anesthetics induced morphologically identical patterns of calcific myonecrosis, formation of scar tissue, and a marked rate of fiber regeneration. However, bupivacaine's effects were constantly more pronounced than those of ropivacaine. These data show that both drugs induce irreversible skeletal muscle damage in a clinically relevant model, and confirm the exceeding rate of myotoxicity of bupivacaine. However, the clinical impact of these long term myotoxic effects still has to be assessed. IMPLICATIONS: In a period of 4 wk after peripheral nerve block, both long-acting local anesthetics, bupivacaine and ropivacaine, produced calcific myonecrosis suggestive of irreversible skeletal muscle damage. In comparison with ropivacaine, however, the extent of bupivacaine-induced muscle lesions was significantly larger. PMID- 16037175 TI - The influence of lumbosacral cerebrospinal fluid volume on extent and duration of hyperbaric bupivacaine spinal anesthesia: a comparison between seated and lateral decubitus injection positions. AB - We designed the present study to examine the influence of lumbosacral cerebrospinal fluid (CSF) volume on the spread and duration of hyperbaric bupivacaine spinal anesthesia when the injection is made with the patient in the lateral position compared with that when the patient is in a seated position. Seventy-four patients undergoing peripheral orthopedic or urogenital surgery with spinal block were enrolled. Lumbosacral CSF volumes were calculated from axial magnetic resonance images. Patients were randomly assigned to 1 of 2 groups: the lateral (L) and seated (S) groups (n = 37 each). Spinal anesthesia (3 mL hyperbaric 0.5% bupivacaine) was administered using a 25-gauge pencil-type needle with the needle aperture directed cephalad and the patient in the lateral decubitus position with the non-operated side up (L group) or with the patient in a seated position (S group). Patients were turned supine immediately after spinal injection (L group) or after remaining seated for 2 min (S group). Statistical correlation coefficients (rho) were assessed using Spearman's rank correlation. There were negative correlations between CSF volume and peak sensory block level in both the L (rho = -0.69, P < 0.0001) and S groups (rho = -0.68, P < 0.0001). In the S group, but not in the L group, CSF volume significantly correlated with onset time of peak sensory block level (rho = -0.48, P = 0.004), and time required for regression to L1-4 (P < 0.05-0.01). We conclude that CSF volume influences the spread of spinal anesthesia with hyperbaric bupivacaine regardless of patient position when the spinal injection is made. CSF volume influenced the duration of spinal sensory anesthesia when the injection was made with the patient in a seated position, but not in the lateral position. IMPLICATIONS: Patient position during the spinal injection does not alter the influence of cerebrospinal fluid (CSF) volume on the spread of hyperbaric bupivacaine spinal anesthesia. However, CSF volume influences the duration of spinal sensory anesthesia when the injection is made with the patient in a seated position, but not in the lateral position. PMID- 16037176 TI - A novel technique for experimental stellate ganglion block in rats. AB - A stellate ganglion block (SGB) is routinely performed in a clinical setting for the treatment of sympathetically maintained pain syndromes. However, the cardiovascular effects of SGB have not been well defined. The purpose of the present study was to develop a new technique of SGB in a rat model. Our new technique of SGB is a posterior percutaneous approach and uses the cartilaginous process of the C7 spinous process as a landmark. Twenty-six Sprague-Dawley female rats were divided into six groups. Group I (n = 4) underwent right sided SGB, Group II (n = 5) underwent left-sided SGB, and Group III (n = 5) underwent bilateral SGB using bupivacaine 0.25%. Three additional sham groups (n = 4 in each group) served as controls to each of the three treatment groups. Ipsilateral eyelid droop (ptosis) was observed in all animals that underwent SGB with bupivacaine. Heart rate decreased significantly for up to 45 min after bilateral SGB compared with control groups. However, this value did not change in rats after unilateral SGB. In 9 additional rats, we evaluated the accuracy of SGB by injecting methylene blue to stain the right (n = 3), left (n = 3), and bilateral SGB (n = 3). At autopsy, 11 of 12 SG were stained post-methylene blue injection. We conclude from our study that our new approach, posterior percutaneous SGB is a reliable technique that can be used for further studies. IMPLICATIONS: We describe a new technique for stellate ganglion block in rats that may be used in future studies to investigate the role of cervical sympathetic nervous system (especially the stellate ganglion) in regulating sympathetically maintained pain and myocardial function. PMID- 16037177 TI - The short-lasting analgesia and long-term antihyperalgesic effect of intrathecal clonidine in patients undergoing colonic surgery. AB - In this study, we investigated the antihyperalgesic effect of clonidine after surgery. Sixty patients undergoing right colic resection were studied. Patients were randomized to receive prior to general anesthesia a 2-mL intrathecal (IT) injection of 300 microg of clonidine or saline, or 10 mg of bupivacaine. General anesthesia was achieved using a target concentration propofol infusion and monitored using bispectral index. Postoperative analgesia was provided by morphine IV given through a patient-controlled analgesia device. Postoperative analgesia was assessed by morphine requirements and visual analog scale pain scores at rest, cough, and movement during the first 72 h. Mechanical hyperalgesia was measured by von Frey filaments. Patients were questioned regarding residual pain at 2 wk,1, 6, and 12 mo. The patient-controlled analgesia morphine requirements were significantly smaller in the IT clonidine group (31.5 +/- 12 versus 91 +/- 25.5 and 43 +/- 15 mg, respectively, in groups clonidine, saline, and bupivacaine: P < 0.05 at 72 postoperative hours). The area of hyperalgesia at 72 h was 3 +/- 5 cm(2) in the clonidine group versus 90 +/- 30 and 35 +/- 20 cm(2) in the saline and bupivacaine groups (P < 0.05). At 6 mo, fewer patients in the clonidine group experienced residual pain than in the saline group (0 of 20 versus 6 of 20, P < 0.05). We conclude that both intraoperative spinal clonidine and bupivacaine improve immediate postoperative analgesia. IT clonidine was, however, more potent than IT bupivacaine to reduce postoperative secondary hyperalgesia. IMPLICATIONS: Spinal clonidine contributes to the reduction of secondary hyperalgesia in patients recovering from abdominal surgery. PMID- 16037178 TI - Intraarticular patient-controlled regional anesthesia after arthroscopically assisted anterior cruciate ligament reconstruction: ropivacaine/morphine/ketorolac versus ropivacaine/morphine. AB - Anterior cruciate ligament reconstruction (ACLR) is associated with moderate to severe postoperative pain. We compared the intraarticular analgesic effects of ropivacaine and morphine with or without ketorolac and the need for rescue IV morphine at rest and during movement in patients undergoing anterior cruciate ligament reconstruction during spinal anesthesia. Thirty-nine patients receiving intraarticular patient-controlled regional analgesia with a 10-mL bolus and a 60 min lockout interval were randomized into 3 groups: the RM group received 0.25% ropivacaine and morphine 0.2 mg/mL; the RMK group received 0.25% ropivacaine, morphine 0.2 mg/mL and ketorolac 1 mg/mL; the P group received saline. Analgesic mixtures were prepared in 100-mL bags and coded. If needed, rescue morphine 2 mg was self-administered IV with 10-min lockout intervals. Pain scores and patient satisfaction were assessed at rest and during movement. There were no significant differences among the groups in pain scores and patient satisfaction. Daily morphine consumption was significantly smaller in the RMK group (8 +/- 8 mg) compared with the RM group (23 +/- 20 mg; P = 0.002) and in both groups compared with control (46 +/- 21 mg; P < 0.001). We conclude that intraarticular patient controlled regional analgesia provides effective pain relief after anterior cruciate ligament reconstruction. The combination of intraarticular ropivacaine, morphine, and ketorolac was superior to control or to a combination of ropivacaine and morphine. IMPLICATIONS: This study showed the feasibility and efficacy of intraarticular patient-controlled regional analgesia technique for pain relief after anterior cruciate ligament reconstruction. The combination of intraarticular ropivacaine, morphine, and ketorolac was superior to control or to a combination of ropivacaine and morphine. PMID- 16037179 TI - Ultrasound guidance for facet joint injections in the lumbar spine: a computed tomography-controlled feasibility study. AB - We conducted this study to develop an ultrasound-guided approach for facet joint injections of the lumbar spine. Five zygapophyseal joints (L1-S1) on each side of 5 embalmed cadavers were examined by ultrasound for a total of 50 examinations. The joint space was demonstrated under ultrasound guidance. The midpoint of the joint space, defined as the middle of its cranio-caudal extension on its dorsal surface, was taken as a reference point, and its position was computed from its depth and lateral distance from the spinous process. Forty-two of 50 approaches could be clearly visualized. Subsequently, these distances were compared to those obtained by computed tomography (CT). To assess the efficacy of ultrasound in the needle placement, all lumbar facet joints were approached in one embalmed cadaver. The exact placement of the needle tips was again evaluated by CT. Ultrasound and CT measurements showed the same mean depth and lateral distance to the reference point, 3.15 +/- 0.5 cm and 1.9 +/- 0.6 cm, respectively. Pearson's coefficient of correlation was 0.86 (P < 0.0001) between ultrasound and CT. All 10 needle tips were within the joint space during simulated facet joint injections. We conclude that ultrasound guidance might be a useful adjunct for facet joint injections in the lumbar spine. IMPLICATIONS: This study was designed to develop an ultrasound-guided approach to the facet joints of the lumbar spine and to assess its feasibility and accuracy by means of a comparison to computed tomography images. The imaging study demonstrated a significant correlation between ultrasound and computed tomography measurements. During simulated facet injection, ultrasound guidance consistently resulted in accurate needle placement. PMID- 16037180 TI - Digital skin blood flow as an indicator for intravascular injection of epinephrine-containing simulated epidural test dose in sevoflurane-anesthetized adults. AB - I designed this study to determine the efficacy of heart rate (HR), systolic blood pressure (SBP), and digital skin blood flow (DSBF) in detecting intravascular injection after a simulated epidural test dose containing 15 mug of epinephrine in sevoflurane-anesthetized adults. In addition, the testing threshold using DSBF was derived. Eighty patients were randomized to receive either 0.5 minimum alveolar anesthetic concentration (MAC) sevoflurane or 1.0 MAC sevoflurane and nitrous oxide in oxygen (n = 40 for each sevoflurane concentration). Each group of patients was further randomized to receive either 3 mL of 1.5% lidocaine containing 15 mug of epinephrine IV or 3 mL of saline IV (n = 20 each). HR, SBP, and DSBF were monitored for 5 min after injection. By using the HR (positive if >or=10 bpm increase) and SBP (positive if >or=15 mm Hg increase) criteria, a positive response rate to epinephrine was 95% for both variables during 0.5 MAC and 90% during 1.0 MAC sevoflurane anesthesia. Injection of the test dose resulted in peak DSBF decrease by 87% +/- 8% and 81% +/- 12% at 52 +/- 10 and 53 +/- 13 s in the sevoflurane 0.5 and 1.0 MAC groups, respectively. Positive DSBF criterion, as determined from peak increases during saline administration, was a decrease in DSBF >or=15%. Using this value, the sensitivity, specificity, positive predictive value, and negative predictive value were 100% in both sevoflurane groups. In conclusion, DSBF was superior to conventional hemodynamic criteria for detection of an intravascular injection of epidural test dose. IMPLICATIONS: This study examined the efficacy of digital skin blood flow to detect an intravascular injection of an epinephrine-containing epidural test dose. This new variable when measured with a laser Doppler flowmeter was superior to conventional hemodynamic criteria during sevoflurane anesthesia. PMID- 16037181 TI - Nerve stimulation for regional anesthesia of the face: use of the blink reflex to confirm the localization of the trigeminal nerve. AB - A blink reflex is a bilateral eyelid closure in response to nociceptive stimuli. We hypothesized that elicitation of a blink by a stimulating needle may enhance performing regional anesthesia of the face. Thirty outpatients undergoing cutaneous surgery of the face were studied. A 2.5-cm insulated beveled stimulating needle was inserted at the emergence of the infraorbital nerve. On elicitation of a bilateral blink with 0.5 mA of stimulating current, bupivacaine 0.5% 3-7 mL was injected. Operative conditions were excellent. Blink response can be used to localize superficial branches of the trigeminal nerve. Elicitation of a blink response during nerve stimulation results in successful trigeminal block. IMPLICATIONS: Elicitation of a blink response during nerve stimulation results in successful trigeminal block. PMID- 16037182 TI - Predictive factors of early postoperative urinary retention in the postanesthesia care unit. AB - Urinary retention is a common postoperative complication associated with bladder overdistension and the risk of permanent detrusor damage. The goal of this study was to determine predictive factors of early postoperative urinary retention in the postanesthesia care unit (PACU). We prospectively collected, in 313 adult patients, variables including age, gender, previous history of urinary tract symptoms, type of surgery and anesthesia, intraoperative administration of anticholinergics, amount of intraoperative fluids, IV morphine titration, and bladder volume on entry to the PACU. For each patient, bladder volume was measured by ultrasound on entry and before discharge from the PACU. Urinary retention was defined as a bladder volume larger than 600 mL with an inability to void within 30 min. Predictive factors were identified by multivariate analysis. The incidence of urinary retention in the PACU was 16%. In the multivariate analysis only the amount of intraoperative fluids (>or=750 mL; P = 0.02; odds ratio = 2.3), age (>or=50 yr; P = 0.008; odds ratio = 2.4), and bladder volume on entry to PACU (>or=270 mL; P = 0.0001; odds ratio = 4.8) were found to independently increase the risk of urinary retention. Considering the clinical impact of undiagnosed postoperative urinary retention, these results suggest systematic evaluation of bladder volume with a portable ultrasound device in the PACU, especially in patients with risk factors. IMPLICATIONS: In this observational study, the ultrasound monitoring of bladder volume in the postanesthesia care unit (PACU) revealed that postoperative urinary retention occurred with an incidence of 16%. Age (>or=50 yr), amount of intraoperative fluid volume (>or=750 mL), and bladder volume on entry to PACU (>or=270 mL) were independent predictive factors for this complication. PMID- 16037183 TI - The influence of postural changes on gastroesophageal reflux and barrier pressure in nonfasting individuals. AB - There is controversy regarding optimal body positioning (i.e., head-up, head down) in awake nonfasting individuals to minimize the risk for pulmonary aspiration of gastric contents as the result of gastroesophageal reflux (GER). In the present study, we investigated GER and intragastric-esophageal barrier pressure by means of multichannel intraluminal impedance measurement and intragastric-esophageal manometry in awake, nonfasting volunteers randomly positioned in a 20 degrees head-up position, the supine position, and a 20 degrees head-down position. No significant difference among positions was found with respect to number of GER episodes per person (0/1/1) or intragastric esophageal barrier pressure (15.6/19.6/19.4 mm Hg). We conclude that specific body positioning is useless in the prophylaxis of GER in awake nonfasting individuals. IMPLICATIONS: Tilting of nonfasting individuals to the head-up or head-down position recommended for prevention of regurgitation of gastric contents does not influence the frequency of gastroesophageal reflux. PMID- 16037184 TI - Intraoperative fluid restriction improves outcome after major elective gastrointestinal surgery. AB - Fluid therapy is one of the most controversial topics in perioperative management. There is continuing debate with regard to the quantity and the type of fluid resuscitation during elective major surgery. However, there are increasing reports of perioperative excessive intravascular volume leading to increased postoperative morbidity and mortality. Recent evidence suggests that judicious perioperative fluid therapy improves outcome after major elective gastrointestinal surgery. The observed benefits may not be solely attributable to crystalloid restriction but also to the use of colloids instead. Some clinically useful guidelines based on the studies discussed in this review include avoidance of deep general anesthesia and elimination of preload for patients who receive epidural analgesia. A balanced approach to fluid management is recommended, with colloids administered to provide hemodynamic stability and maintain urine output of 0.5 mL x kg(-1) x h(-1) and crystalloids administered only for maintenance. In addition, blood loss may be replaced with colloid on a volume-to-volume basis. Furthermore, predetermined algorithms that suggest replacement of third space losses and losses through diuresis are unnecessary. Significant reduction in crystalloid volume can be achieved without encountering intraoperative hemodynamic instability or reduced (i.e., < 0.5 mL x kg(-1) x h(-1)) urinary output just by avoiding replacement of third space losses and preloading. Finally, there is a need for well-controlled studies in a well-defined patient population using clear criteria or end-points for perioperative fluid therapy. PMID- 16037185 TI - A simple method to identify an external vaporizer leak (the "SNIFF" method). PMID- 16037187 TI - Pro-drugs in in vitro experiments. PMID- 16037189 TI - Lidocaine toxicity in volunteer subjects undergoing awake fiberoptic intubation. PMID- 16037190 TI - Prolonged postoperative disorientation after methylene blue infusion during parathyroidectomy. PMID- 16037191 TI - Anesthetic management using echocardiography for surgery of lower extremity in a patient with Ebstein's malformation. PMID- 16037193 TI - Intrathecal morphine for postpartum bilateral tubal ligation. PMID- 16037195 TI - Ultrasound evidence of intraneural injection. PMID- 16037197 TI - Awareness during anesthesia. PMID- 16037198 TI - Existential distress and palliative sedation. PMID- 16037200 TI - Intraoperative detection of tracheobronchial placement of a nasogastric tube: a new observational method. PMID- 16037201 TI - Fire during thoracotomy: a need to control the inspired oxygen concentration. PMID- 16037203 TI - Reduction in requirement of propofol during combined epidural (bupivacaine) and general anesthesia guided by bispectral index. PMID- 16037204 TI - Anesthesia for Freeman-Sheldon syndrome using a folded laryngeal mask airway. PMID- 16037206 TI - Postischemic nitrous oxide alone versus intraischemic nitrous oxide in the presence of isoflurane: what it may change for neuroprotection against cerebral stroke in the rat. PMID- 16037207 TI - Inhibition of a vaccine-induced anti-tumor B cell response by soluble protein antigen in the absence of continuing T cell help. AB - DNA vaccination can elicit the production of anti-tumor antibodies, thus obviating the need to continuously administer passive antibody. This vaccination strategy is particularly important where antibodies have proven to be effective anti-tumor agents. To amplify antibody responses against weak tumor antigens, we previously designed DNA-fusion vaccines incorporating tumor sequences linked to microbial genes. By using a safe idiotypic (Id) antigen from a B cell tumor fused to a fragment C (FrC) sequence from tetanus toxin, we induced both anti-Id and anti-FrC antibodies. It was important to determine whether the antigen itself, either injected or released from residual tumor cells, would boost the antibody response. Id protein not only failed to boost the response, but permanently and rapidly inhibited it by ablating Id-specific memory B cells. In contrast, an Id protein-FrC conjugate boosted both Id-specific and FrC-specific responses. Strikingly, the depletion of CD4+ T cells converted the Id protein-FrC conjugate vaccine into an inhibitor. These findings support the hypothesis that the activation of memory B cells by a DNA vaccine encoding a protein antigen, in the presence of the protein itself, depends completely on T cell help. Furthermore, by using knockout mice, we have shown that inhibition of the Id-specific memory B cells by the Id protein is largely independent of the FcgammaRIIB and, hence, independent of immune complexes. The principles revealed by using a DNA vaccine have implications for all cancer vaccines designed to induce and maintain antibody responses against weak autologous tumor antigens. PMID- 16037208 TI - Effective function annotation through catalytic residue conservation. AB - Because of the extreme impact of genome sequencing projects, protein sequences without accompanying experimental data now dominate public databases. Homology searches, by providing an opportunity to transfer functional information between related proteins, have become the de facto way to address this. Although a single, well annotated, close relationship will often facilitate sufficient annotation, this situation is not always the case, particularly if mutations are present in important functional residues. When only distant relationships are available, the transfer of function information is more tenuous, and the likelihood of encountering several well annotated proteins with different functions is increased. The consequence for a researcher is a range of candidate functions with little way of knowing which, if any, are correct. Here, we address the problem directly by introducing a computational approach to accurately identify and segregate related proteins into those with a functional similarity and those where function differs. This approach should find a wide range of applications, including the interpretation of genomics/proteomics data and the prioritization of targets for high-throughput structure determination. The method is generic, but here we concentrate on enzymes and apply high-quality catalytic site data. In addition to providing a series of comprehensive benchmarks to show the overall performance of our approach, we illustrate its utility with specific examples that include the correct identification of haptoglobin as a nonenzymatic relative of trypsin, discrimination of acid-d-amino acid ligases from a much larger ligase pool, and the successful annotation of BioH, a structural genomics target. PMID- 16037209 TI - Contributions of past and present human generations to committed warming caused by carbon dioxide. AB - We developed a highly simplified approach to estimate the contributions of the past and present human generations to the increase of atmospheric CO(2) and associated global average temperature increases. For each human generation of adopted 25-year length, we use simplified emission test cases to estimate the committed warming passed to successive children, grandchildren, and later generations. We estimate that the last and the current generation contributed approximately two thirds of the present-day CO(2)-induced warming. Because of the long time scale required for removal of CO(2) from the atmosphere as well as the time delays characteristic of physical responses of the climate system, global mean temperatures are expected to increase by several tenths of a degree for at least the next 20 years even if CO(2) emissions were immediately cut to zero; that is, there is a commitment to additional CO(2)-induced warming even in the absence of emissions. If the rate of increase of CO(2) emissions were to continue up to 2025 and then were cut to zero, a temperature increase of approximately 1.3 degrees C compared to preindustrial conditions would still occur in 2100, whereas a constant-CO(2)-emissions scenario after 2025 would more than double the 2100 warming. These calculations illustrate the manner in which each generation inherits substantial climate change caused by CO(2) emissions that occurred previously, particularly those of their parents, and shows that current CO(2) emissions will contribute significantly to the climate change of future generations. PMID- 16037210 TI - Redundant control of the Caenorhabditis elegans sperm/oocyte switch by PUF-8 and FBF-1, two distinct PUF RNA-binding proteins. AB - PUF proteins control both growth and differentiation in the C. elegans germ line. These conserved RNA-binding proteins inhibit expression of target mRNAs, either by repressing translation or promoting degradation. Previous studies showed that PUF-8, a PUF protein with striking similarity to human Pumilio, prevents return of primary spermatocytes to the mitotic cell cycle [Subramaniam, K. & Seydoux, G. (2003) Curr. Biol. 13, 134-139]. We now report that PUF-8 is also critical for the hermaphrodite sperm/oocyte switch. Most puf-8 mutant hermaphrodites make both sperm and oocytes and are self-fertile, but some make a vast excess of sperm and fail to switch into oogenesis. This puf-8 defect is dramatically enhanced by removal of another puf gene called fbf-1: all fbf-1 puf-8 double mutants fail in the hermaphrodite sperm/oocyte switch. Therefore, puf-8 and fbf-1 act redundantly to control this decision. Epistasis analyses place puf-8 and fbf-1 upstream of fog-2, a gene near the top of the germ-line sex determination pathway. Furthermore, the abundance of FOG-2 increases dramatically in the distal region of fbf-1 puf-8 double mutants. We suggest that PUF-8 and FBF-1 may control fog-2 expression, and that the sperm/oocyte decision occurs in the distal germ line. PMID- 16037211 TI - Modulation of disease, T cell responses, and measles virus clearance in monkeys vaccinated with H-encoding alphavirus replicon particles. AB - Measles remains a major worldwide problem partly because of difficulties with vaccination of young infants. New vaccine strategies need to be safe and to provide sustained protective immunity. We have developed Sindbis virus replicon particles that express the measles virus (MV) hemagglutinin (SIN-H) or fusion (SIN-F) proteins. In mice, SIN-H induced high-titered, dose-dependent, MV neutralizing antibody after a single vaccination. SIN-F, or SIN-H and SIN-F combined, induced somewhat lower responses. To assess protective efficacy, juvenile macaques were vaccinated with a single dose of 10(6) or 10(8) SIN-H particles and infant macaques with two doses of 10(8) particles. A dose of 10(8) particles induced sustained levels of high-titered, MV-neutralizing antibody and IFN-gamma-producing memory T cells, and most monkeys were protected from rash when challenged with wild-type MV 18 months later. After challenge, there was a biphasic appearance of H- and F-specific IFN-gamma-secreting CD4+ and CD8+ T cells in vaccinated monkeys, with peaks approximately 1 and 3-4 months after challenge. Viremia was cleared within 14 days, but MV RNA was detectable for 4-5 months. These studies suggest that complete clearance of MV after infection is a prolonged, phased, and complex process influenced by prior vaccination. PMID- 16037212 TI - Alterations in the alpha2 isoform of Na,K-ATPase associated with familial hemiplegic migraine type 2. AB - A number of missense mutations in the Na,K-ATPase alpha2 catalytic subunit have been identified in familial hemiplegic migraine with aura. Two alleles (L764P and W887R) showed loss-of-function, whereas a third (T345A) is fully functional but with altered Na,K-ATPase kinetics. This study describes two additional mutants, R689Q and M731T, originally identified by Vanmolkot et al. [Vanmolkot, K. R., et al. (2003) Ann. Neurol. 54, 360-366], which we show here to also be functional and kinetically altered. Both mutants have reduced catalytic turnover and increased apparent affinity for extracellular K(+). For both R689Q and M731T, sensitivity to vanadate inhibition is decreased, suggesting that the steady-state E(1) <==> E(2) poise of the enzyme is shifted toward E(1). Whereas the K'(ATP) is not affected by the R689Q replacement, the M731T mutant has an increase in apparent affinity for ATP. Analysis of the structural changes effected by T345A, R689Q, and M731T mutations, based on homologous replacements in the known crystal structure of the sarcoplasmic reticulum Ca-ATPase, provides insights into the molecular bases for the kinetic alterations. It is suggested that the disease phenotype is the consequence of lowered molecular activity of the alpha2 pump isoform due to either decreased K(+) affinity (T345A) or catalytic turnover (R689Q and M731T), thus causing a delay in extracellular K(+) clearance and/or altered localized Ca(2+) handling/signaling secondary to reduced activity in colocalized Na(+)/Ca(2+) exchange. PMID- 16037213 TI - Activationless electron transfer through the hydrophobic core of cytochrome c oxidase. AB - Electron transfer (ET) within proteins occurs by means of chains of redox intermediates that favor directional and efficient electron delivery to an acceptor. Individual ET steps are energetically characterized by the electronic coupling V, driving force DeltaG, and reorganization energy lambda. lambda reflects the nuclear rearrangement of the redox partners and their environment associated with the reactions; lambda approximately 700-1,100 meV (1 eV = 1.602 x 10(-19) J) has been considered as a typical value for intraprotein ET. In nonphotosynthetic systems, functional ET is difficult to assess directly. However, using femtosecond flash photolysis of the CO-poised membrane protein cytochrome c oxidase, the intrinsic rate constant of the low-DeltaG electron injection from heme a into the heme a(3)-Cu(B) active site was recently established at (1.4 ns)(-1). Here, we determine the temperature dependence of both the rate constant and DeltaG of this reaction and establish that this reaction is activationless. Using a quantum mechanical form of nonadiabatic ET theory and common assumptions for the coupled vibrational modes, we deduce that lambda is <200 meV. It is demonstrated that the previously accepted value of 760 meV actually originates from the temperature dependence of Cu(B)-CO bond breaking. We discuss that low-DeltaG, low-lambda reactions are common for efficiently channeling electrons through chains that are buried inside membrane proteins. PMID- 16037214 TI - Slc7a11 gene controls production of pheomelanin pigment and proliferation of cultured cells. AB - In mammals, >100 genes regulate pigmentation by means of a wide variety of developmental, cellular, and enzymatic mechanisms. Nevertheless, genes that directly regulate pheomelanin production have not been described. Here, we demonstrate that the subtle gray (sut) mouse pigmentation mutant arose by means of a mutation in the Slc7a11 gene, encoding the plasma membrane cystine/glutamate exchanger xCT [Kanai, Y. & Endou, H. (2001) Curr. Drug Metab. 2, 339-354]. A resulting low rate of extracellular cystine transport into sut melanocytes reduces pheomelanin production. We show that Slc7a11 is a major genetic regulator of pheomelanin pigment in hair and melanocytes, with minimal or no effects on eumelanin. Furthermore, transport of cystine by xCT is critical for normal proliferation, glutathione production, and protection from oxidative stress in cultured cells. Thus, we have found that the Slc7a11 gene controls the production of pheomelanin pigment directly. Cells from sut mice provide a model for oxidative stress-related diseases and their therapies. PMID- 16037216 TI - Historical perspective of the Maillard reaction in food science. AB - Maillard's paper of 1912 describing the reaction between amino acids and sugars is both innovative and visionary. It provides original and still-valuable data on the chemistry of a new reaction and foresees its involvement in many scientific and biological domains, even in human pathology. This paper was ignored by the scientific community until 1941. In 1948 the Maillard reaction was definitely recognized as being responsible for the browning and loss of nutritive value of heated milk powders. There was then a continuous increase in papers on the chemistry of this complex reaction to identify its various pathways: in food science, to evaluate the influence of reaction parameters (pH, T degrees , time, sugar reactivity, concentration of the reagents, water activity, glass transition temperature) on the evolution of the reaction and on changes in food quality; in nutrition, to quantify the loss of bioavailability of essential amino acids; on the metabolism of the reaction products and on the physiological effects of the ingested Maillard reaction products. The significant scientific advances and the key persons and pioneers who contributed much to the understanding of the Maillard reaction are presented. The food industry is directly concerned with the occurrence of this reaction in processed foods and contributed significantly by its own research to understanding the phenomena and to optimizing the processes and conditions of food preparation in order to preserve the nutritional, safety, and organoleptic qualities of foods. PMID- 16037215 TI - A high frequency of overlapping gene expression in compacted eukaryotic genomes. AB - The gene density of eukaryotic nuclear genomes is generally low relative to prokaryotes, but several eukaryotic lineages (many parasites or endosymbionts) have independently evolved highly compacted, gene-dense genomes. The best studied of these are the microsporidia, highly adapted fungal parasites, and the nucleomorphs, relict nuclei of endosymbiotic algae found in cryptomonads and chlorarachniophytes. These systems are now models for the effects of compaction on the form and dynamics of the nuclear genome. Here we report a large-scale investigation of gene expression from compacted eukaryotic genomes. We have conducted EST surveys of the microsporidian Antonospora locustae and nucleomorphs of the cryptomonad Guillardia theta and the chlorarachniophyte Bigelowiella natans. In all three systems we find a high frequency of mRNA molecules that encode sequence from more than one gene. There is no bias for these genes to be on the same strand, so it is unlikely that these mRNAs represent operons. Instead, compaction appears to have reduced the intergenic regions to such an extent that control elements like promoters and terminators have been forced into or beyond adjacent genes, resulting in long untranslated regions that encode other genes. Normally, transcriptional overlap can interfere with expression of a gene, but these genomes cope with high frequencies of overlap and with termination signals within expressed genes. These findings also point to serious practical difficulties in studying expression in compacted genomes, because many techniques, such as arrays or serial analysis of gene expression will be misleading. PMID- 16037217 TI - The discovery of glycated hemoglobin: a major event in the study of nonenzymatic chemistry in biological systems. AB - Glycated hemoglobins are minor components of human hemoglobin (Hb). These are formed nonenzymatically by condensation of glucose or other reducing sugars with alpha- and beta-chains of hemoglobin A. The subfraction HbA1c, a nonenzymatic glycation at the amino-terminal valines of the beta-chain, was identified by the author in the 1960s as a minor "abnormal fast-moving hemoglobin band" in diabetic patients during routine screening for hemoglobin variants. This finding later turned out to be an important biomolecular marker with clinical and pathological applications. Measurement of HbA1c in diabetic patients is an established procedure for evaluating long-term control of diabetes, and the introduction of this measurement represents an outstanding contribution to the quality of care of diabetic patients in this century. More importantly, HbA1c is the first example of in vivo nonenzymatic glycation of proteins, and its discovery opened new and still-growing avenues of research on Maillard reactions in biological systems, including the concept of advanced glycation/lipoxidation end products (AGEs/ALEs) and the development of diabetic complications and various diseases associated with aging. Although interest in the Maillard reaction is growing rapidly, much remains to be done in this field, including detection and characterization of all in vivo AGEs/ALEs, development and clinical applications of AGE inhibitors and breakers, as well as investigations into the possible roles of the Maillard reaction in regulatory biology and carcinogenesis. PMID- 16037218 TI - Taste-active maillard reaction products: the "tasty" world of nonvolatile maillard reaction products. AB - This study was done to obtain greater insight into the structures and sensory activities of those tastants that are not present in foods per se, but are generated during food processing by Maillard-type reactions from carbohydrates and amino acids and thus remain unknown. In order to rank the tastants according to their relative taste impact and to identify the key tastants generated during thermal food processing, the so-called taste dilution analysis (TDA), which uses the human tongue as a biosensor for tastants, was applied to heated, intensely bitter tasting binary mixtures of glucose or xylose and proline or alanine, respectively. This screening technique led to the identification of previously unknown taste compounds, among which intensely bitter tastants such as quinizolate and homoquinizolate, a pungent-tasting pyranopyranone, cyclopentenone derivatives exhibiting a physiological cooling effect, as well as a taste enhancing pyridinium betaine named alapyridaine will be presented. PMID- 16037219 TI - Current status of acrylamide research in food: measurement, safety assessment, and formation. AB - Relatively high amounts of acrylamide have recently been reported in carbohydrate rich foods under low moisture conditions. This sparked intensive investigations into acrylamide, encompassing its occurrence, chemistry, and toxicology/potential health risk in the human diet. Robust and efficient analytical methods have been developed to obtain reliable quantitative data. Recent epidemiological studies failed to evidence an association of cancer incidence and dietary acrylamide exposure. The link of acrylamide in foods to Maillard-type reactions and, in particular, to asparagine has been a major step in elucidating feasible chemical formation routes. Decarboxylation of the Schiff base derived from asparagine and a carbonyl reactant plays a key role in acrylamide formation leading to azomethine ylide intermediates, which offer an attractive mechanistic explanation of the acrylamide amounts found experimentally, including the fact that acrylamide is preferentially formed in the presence of fructose, as compared to glucose or alpha-dicarbonyls. However, the physical state of the reaction system may also affect acrylamide formation by influencing molecular mobility, particularly under low moisture conditions. Current research deals mainly with mitigation studies to reduce acrylamide during food processing and the role of water. PMID- 16037220 TI - Oxidative pyrolysis and postpyrolytic derivatization techniques for the total analysis of maillard model systems: investigation of control parameters of maillard reaction pathways. AB - Factors that regulate various pathways of Maillard reaction leading to aroma, color, or carcinogen generation have not been identified, due to the difficulties associated with analyzing complex reaction mixtures. In particular, the role played by oxidation in directing aromagenic, chromogenic, or carcinogenic pathways is not well understood. In order to overcome the analytical difficulties, novel Py-GC/MS-based methodologies were developed to analyze volatile and nonvolatile residues of Maillard reaction products generated from the same model system under air or helium atmosphere. The analysis of nonvolatiles was achieved through a postpyrolytic in situ derivatization technique using hexamethyldisilazane, and pyrolysis under air was achieved through modification of the GC equipped with sample concentration trap to allow gas stream switching and subsequent isolation of the pyrolysis chamber from the analytical stream. In this approach label incorporation from the starting materials can be observed in both volatile and nonoxidative conditions for mechanistic studies. In addition, monitoring of redox potentials, oxygen consumption, and color generation of relevant model systems over time were also carried out at different temperatures. The data collected have indicated that perturbation in the redox potential of Maillard model systems by external (oxidizing conditions) or internal (formation of reductones) factors can alter the balance among the four critically important groups of precursors: alpha dicarbonyl, alpha-hydroxycarbonyl, 2-aminocarbonyls, and 2-(amino acid)-carbonyl compounds and hence control the relative importance of aromagenic versus chromogenic pathways. PMID- 16037221 TI - Identification of N7-(1-carboxyethyl)-arginine, a novel posttranslational protein modification of arginine formed at high hydrostatic pressure. AB - An aqueous solution of Nalpha-acetylarginine (0.1 mol/L) and glucose (0.3 mol/L) in 0.5 mol/L 3-[N-morpholino]propanesulfonic acid (5 mL; pH 7.0) was incubated at 110 degrees C for 2 h with application of high hydrostatic pressure (HHP). After enzymic deacetylation, the arginine modifications formed were separated by ion exchange chromatography. Besides N5-(4,5-dihydro-4-methyl-5-oxo-1H-imidazol-2-yl) L-ornithine, the novel arginine modification N7-(1-carboxyethyl)-arginine was identified based on NMR and LC/MS measurements and synthesis-in particular, in samples reacted at a pressure of 600 MPa. As compared to samples treated at normal pressure, both compounds were formed in higher amounts at increased pressure. PMID- 16037222 TI - Labeling studies on the formation pathway of Nepsilon-carboxymethyllysine in maillard-type reactions. AB - Nepsilon-carboxymethyllysine (CML) is among the most often studied advanced glycation end products (AGEs) in foods and medicine, and several pathways have been suggested for its formation. Based on the results of model studies using glucose labeled with carbon-13 in different positions, a new pathway is proposed for CML formation without involving free glyoxal or autoxidation reactions. PMID- 16037223 TI - Elucidation of chemical pathways in the maillard reaction by 17O-NMR spectroscopy. AB - 17O-NMR spectroscopy was employed as an innovative method to help understand mechanistic pathways in sugar fragmentation. Elucidation of reaction mechanisms to final Maillard end products was achieved by starting from specific intermediates obtained by synthesis, such as 1-deoxy-D-erythro-hexo-2,3-diulose. This alpha-dicarbonyl was thermally treated in the presence of 17O-enriched water under alkaline conditions. The reaction products were monitored by 17O-NMR spectroscopy and their structures corroborated by complementary techniques. For the first time, evidence is shown for the direct formation of acetic acid from 1 deoxy-D-erythro-hexo-2,3-diulose by an oxidative alpha-dicarbonyl cleavage and incorporation of a 17OH group into the acetic acid released as sugar fragment. PMID- 16037224 TI - The effect of reaction conditions on the origin and yields of acetic acid generated by the maillard reaction. AB - The effect of the reaction conditions on the origin and yields of acetic acid from glucose was studied in the system containing equimolar concentrations of 13C labeled glucose and glycine. Acetic acid was quantified by GC-MS using isotope dilution assay. The beta-dicarbonyl cleavage of 1-deoxyhexo-2,4-diulose is proposed to be a major pathway leading to the formation of acetic acid in the glucose-based Maillard reaction systems under food processing conditions. Acetic acid was built up from all six carbon atoms of glucose. The relative distribution of acetic acid was independent of the reaction time. Temperature and pH had only small effects. PMID- 16037225 TI - Formation of mutagenic/carcinogenic heterocyclic amines under moderate conditions. AB - More than 10 kinds of heterocyclic amines (HCAs), showing mutagenic and carcinogenic potency, have been isolated from cooked fish and meat. But many researchers say that the contribution ratio of HCAs to human cancer is very low. Our purpose in this experiment was to investigate the possibility of the formation of HCAs under moderate conditions, including in vivo. A mixture of d glucose, creatinine, and amino acid such as glycine, methionine, threonine, and proline was dissolved in phosphate-buffered solution (pH7.4) and incubated at 37 degrees C, 50 degrees C, 128 degrees C. At an appropriate time, an aliquot of the reaction solution was treated with blue cotton. HCAs were separated from the blue cotton by elution with 2% ammoniacal methanol. The eluates were submitted to the Ames test, the micronucleus test for determination of mutagenicity, and also LC MS analysis for the detection of HCAs. Nonadsorbates to blue cotton were treated with dichloromethane and then subjected to the mutagenicity test. In the Ames test, the mutagenic activity of the reaction mixture increased with an increase of the reaction temperature. The HCA fraction from 50 degrees C incubated solution showed high frequency in the micronucleus test using HepG2 cells. The dichloromethane fractions contained other type of mutagens different from HCAs. In HCA fractions, IQ, MeIQx, 4,8-DiMeIQx, and 7,8-DiMeIQ were identified. It is said that the heating process is an essential factor in the formation of HCAs. But our experiment shows that HCAs are produced not only in the cooking process, but also in moderate conditions such as 37 degrees C and 50 degrees C. PMID- 16037226 TI - Maillard reactions of ribose 5-phosphate and amino acids. AB - An important metabolite in nucleotide synthesis, ribose 5-phosphate (R5P) undergoes Maillard reactions at a rate significantly faster than most common sugars and sugar phosphates of its type. At pH = 8 and 37 degrees C, R5P reacts with amino acids to generate UV-absorbing compounds within hours and brownish compounds within a day. The increased rate can be attributed to the strategic location of the 5-phosphate group near the reactive C1 center. As expected, the rate of reaction is at least partially related to pKa of the attacking amine and it follows first-order kinetics in respect to amine (in the case of glycine). In terms of producing 280 nm absorbing compounds, the reaction order in respect to R5P was 1.5. The reaction mechanism appears to proceed through a 1-deoxyribosome intermediate. In reactions with proteins, R5P exhibits an ability to promote cross-linking. PMID- 16037227 TI - Protein cross-linking in food. AB - The aims of this paper are (1) to probe the relationship between molecular structure and protein cross-linking ability for a range of small molecules; (2) to establish whether this relationship holds within a food matrix; and (3) to test the impact of Maillard cross-linking on food functionality, particularly texture, in wheat- and soy-based food systems. A variety of molecules were obtained, either commercially or via organic synthesis. Cross-linking ability was tested using our standard model system, employing ribonuclease A and analyzing the results by SDS-PAGE. Molecules of varying reactivity were tested in wheat- and soy-based products, and the changes in functionality were correlated with changes in protein cross-linking. No simple relationship was found between molecular structure and ability to cross-link ribonuclease. Only the most reactive reagents were able to cross-link within the food matrix. Nevertheless, a low degree of cross-linking was shown to have significant consequences on the properties of wheat- and soy-based foods, suggesting that the Maillard reaction may represent a means to control food texture. PMID- 16037228 TI - Chemistry and biological effects of melanoidins and glyceraldehyde-derived pyridinium as advanced glycation end products. AB - Blue pigments (blue-M1 and blue-M2) and red pigments (red-M1 and red-M2) were generated in a xylose-glycine reaction system. Blue-M2 was identified as an addition compound of di-xylulose-glycine to blue-M1 that involved two pyrrolopyrrole structures. We identified red pigments as isomers of addition compounds of xylulose-glycine to the condensed compound between pyrrolopyrrole-2 carbaldehyde and pyrrole-2-carbaldehyde. These pigments have polymerizing activity, suggesting that they are important Maillard reaction intermediates through the formation of melanoidins. Melanoidins induced IFN-gamma and IL-12 expression in spleen cells exposed to allergen and in macrophages, respectively. These findings suggest that melanoidins have a suppressive effect on allergic reaction as a novel physiological effect. On the other hand, we identified a glyceraldehyde-derived advanced glycation end product (AGE) formed from glyceraldehyde and N-acetylarginine as well as glyceraldehyde-derived pyridinium (GLAP) in physiological conditions. The AGE was identified as 5-methylimidazoline 4-one (MG-H1), which has been reported to be formed from arginine and methylglyoxal. GLAP, which induced reactive oxygen species (ROS) production in HL 60 cells, is supposed to be a toxic AGE, while MG-H1 is a nontoxic AGE. PMID- 16037229 TI - Dicarbonyl intermediates in the maillard reaction. AB - The complexity of the Maillard reaction arises partly from multiple fragmentation reactions of the sugar moiety, constituting branch points in the reaction progress and establishing many parallel reaction pathways. Reactive intermediates produced by these processes are often alpha-oxoaldehydes. The formation of alpha oxoaldehydes enhances and redirects glycating activity in the Maillard reaction since alpha-oxoaldehydes are up to 20,000-fold more reactive than glucose in glycation processes and are predominantly arginine-directed glycating agents. alpha-Oxoaldehydes bypass a requirement for a fructosamine precursor in the formation of advanced glycation end products (AGEs) since alpha-oxoaldehydes react with proteins (also nucleotides and basic phospholipids) to form AGEs directly. The major AGE formed from alpha-oxoaldehydes is generally a hydroimidazolone with other products-although for glyoxal, N(omega) carboxymethylarginine is a major product. alpha-Oxoaldehyde formation also occurs in the absence of an amine substrate, particularly during heat processing of sugar solutions and lipid peroxidation processes-in the latter case, the glycation adducts are advanced lipoxidation products (ALEs). Hydroimidazolones are quantitatively important AGEs in cellular and extracellular proteins in physiological systems. Hydroimidazolone free adducts are liberated by cellular proteolysis and digestion. They are released into blood plasma for urinary excretion. Modification of arginine residues by alpha-oxoaldehydes may be particularly damaging since arginine residues have high-frequency occurrence in ligand and substrate recognition sites in receptor and enzyme active sites. Along with fructosamine formation, alpha-oxoaldehyde intermediates of the Maillard reaction represent a major source of damage to the proteome and genome. PMID- 16037230 TI - Ornithine is a novel amino acid and a marker of arginine damage by oxoaldehydes in senescent proteins. AB - Long-lived proteins undergo age-related postsynthetic modifications by glycation and advanced glycation end products (AGEs), which destabilize them by altering their conformation and charge. It was accidentally discovered that ornithine (orn) increased with age in acid hydrolyzates of human skin collagen and lens crystallins which led us to investigate the source of orn. Here, we detected such modifications of orn in these proteins. Acid hydrolysis of arginine (arg)-base AGE standards produced orn at different yields. The data provide unequivocal evidence for the in vivo formation of orn and its own AGEs in aging proteins, and suggest that arg-based AGEs serve as precursors of orn. PMID- 16037231 TI - Formation of alpha-aminoadipic and gamma-glutamic semialdehydes in proteins by the maillard reaction. AB - Recent research has demonstrated that nonenzymatic glycation (the Maillard reaction) lead to the formation of carbonyl groups and advanced glycation end products (AGEs) in proteins. Such oxidative modifications are a major contributing factor to diabetic complications and aging. alpha-Aminoadipic semialdehyde (AAS) and gamma-glutamic semialdehyde (GGS) have been identified as the major carbonyl products in oxidized proteins both in vitro and in vivo. AAS is an oxidative deamination product of lysine residue, while GGS originates from arginine and proline residues. To evaluate oxidative damage to proteins by the Maillard reaction, we developed a method of detecting AAS and GGS by high performance liquid chromatography (HPLC). The aldehydic residues in proteins were derivatized by reductive amination with NaCNBH3 and p-aminobenzoic acid (ABA), a fluorescence regent. After acid hydrolysis of the ABA-derivatized protein, ABA AAS and ABA-GGS were measured by fluorometric HPLC. Thus, AAS and GGS could be detected in various proteins such as human plasma protein using the present method. Accumulation of both aldehydic residues was observed in oxidized proteins by reactive oxygen species. Furthermore, AAS and GGS were markedly formed in the incubation of BSA with ascorbic acid. The formation of both aldehydic residues was also observed in the incubation of BSA with 100 mM glucose or 1.0 mM methylglyoxal in the absence and presence of 100 microM Fe3+ for 2 weeks. These results suggest that the Maillard reaction can contribute to the formation of AAS and GGS in vivo. PMID- 16037232 TI - Methylglyoxal can modify GAPDH activity and structure. AB - The activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) can play an important role in regulating multiple upstream pathways relating to the development of diabetic complications. GAPDH can be modified by a number of metabolic factors, including oxidative and glycation products. To study the effect of glycation on GAPDH we have measured GAPDH structure and activity after exposure of the enzyme to the potent alpha dicarbonyl sugar methylglyoxal (MG). Rabbit GAPDH was incubated with 10-1000 microM MG for 96 hours, and enzyme activity was measured at intervals by a spectrophotometric assay. Isoelectric focusing of purified and cellular GAPDH was performed with a PROTEAN IEF system and the bands visualized by Western blotting. The mass of glycated and native GAPDH was determined by MALDI with a Applied Biosystems Voyager System 6235. GAPDH activity (at 96 h) was decreased by 20% with 1.0 micromolar MG and showed progressively greater suppression of activity with increasing concentrations up to 1 mM, where activity was decreased by 97%. Reduction in GAPDH activity was rapidly decreasing by 69.2% by two hours with 1 mM MG. IEF showed an isoelectric point (IEP) of 8.5 for native GAPDH, while measurable changes were seen with modification by MG levels of 1 mM (IEP 7.5) and 50 microM (IEP 8.0). With MALDI, GAPDH mass increased from 36.012 kDa to 37.071 after exposure to 50 microM MG and to 40.625 following 1 mM MG. This indicates addition of 12.75 and 55.6 MG residues, respectively, to GAPDH. GAPDH can be modified by methylglyoxal intracellular concentrations close to those previously observed in vivo, with measurable changes in isoelectric point and mass. These modifications can lead to decreased enzyme activity, suggesting that conditions associated with elevated intracellular MG could modify GAPDH activity in vivo. PMID- 16037233 TI - Effect of glucose concentration on formation of AGEs in erythrocytes in vitro. AB - Posttranslational modifications, such as advanced glycoxidation and lipoxidation end products (AGE/ALEs), are implicated in the pathogenesis of diabetic complications and atherosclerosis. Recent studies have demonstrated that AGE/ALEs are generated not only in extracellular matrix proteins, but also in intracellular proteins from metabolic intermediates. In this study we investigate the effect of glucose concentration on the formation of the AGE/ALEs, Nepsilon (carboxymethyl)lysine (CML), Nepsilon-(carboxyethyl)lysine (CEL), S (carboxymethyl)cysteine (CMC), and S-(2-succinyl)cysteine (2SC) in erythrocytes as a function of glucose concentration. Human erythrocytes (10% hematocrit) were incubated in Dulbecco's modified Eagle's medium (DMEM) containing 5 mM or 30 mM glucose for 5 days at 37 degrees C. Globin was recovered by precipitation with 0.25 M HCl in acetone. Following acid hydrolysis, amino acids were converted to their trifluoroacetyl methyl ester derivatives and analyzed by GC/MS/MS. The CML and CEL content of globin increased in a time- and glucose-dependent manner and also increased 1.3- and 1.8-fold, respectively, in incubations containing 30 mM glucose; whereas CMC and 2SC content did not change during the five-day incubations. Furthermore, CEL content of globin in erythrocytes incubated with 30 mM was the highest in the other AGEs, indicating that methylglyoxal may play a major role in AGE formation in erythrocytes. The erythrocyte system should be useful for cellular screening of the efficacy of inhibitors of AGE/ALE formation. PMID- 16037234 TI - Methylglyoxal induces apoptosis through oxidative stress-mediated activation of p38 mitogen-activated protein kinase in rat Schwann cells. AB - Although recent studies have suggested the potential involvement of apoptotic cell death in the development of diabetic neuropathy, the precise mechanism remains to be elucidated. On the other hand, it is known that the formation of methylglyoxal (MG), a highly reactive dicarbonyl compound, is accelerated under diabetic conditions through several glucose-related metabolisms including the glycation reaction. We found that MG was capable of inducing apoptosis in peripheral nerve-derived Schwann cells (SCs) in a time- and dose-dependent manner, accompanied by a reduction of intracellular glutathione content. Furthermore, MG induced phosphorylation of MKK3/MKK6, an upstream molecule in the p38 MAPK pathway. N-acetyl-L-cysteine, an antioxidant, successfully suppressed the activity of the p38 MAPK signaling pathway along with the inhibition of apoptosis, indicating the involvement of oxidative stress in the MG-induced apoptosis via the p38 MAPK pathway. These results suggest a possible contribution of glucose-derived MG to the development of diabetic neuropathy by injuring the cellular constituent of the peripheral nerve system, such as SCs, in the hyperglycemic milieu. PMID- 16037235 TI - Dicarbonyl stress and apoptosis of vascular cells: prevention by alphaB crystallin. AB - Methylglyoxal (MGO) is an alpha-dicarbonyl compound produced from triose phosphate intermediates of glycolysis. It reacts rapidly with proteins to produce advanced glycation products. We have studied the effect of MGO modification of fibronectin on retinal capillary cell viability. Our studies show that pericytes grown on MGO-modified fibronectin (FN) undergo enhanced apoptosis through the p38MAPK-mediated oxidative pathway and that alphaB-crystallin, a stress protein present in pericytes, can protect them from MGO-mediated apoptosis. Our studies with vascular endothelial cells show that hyperglycemia-induced apoptosis is inhibited by overexpression of alphaB-crystallin. These observations suggest a novel role of alphaB-crystallin in hyperglycemia-mediated damage to vascular cells in diabetes. PMID- 16037236 TI - Phototransformations of advanced glycation end products in the human eye lens due to ultraviolet A light irradiation. AB - Previous studies from this laboratory have shown that ultraviolet A (UVA) light can bleach the yellow advanced glycation end products (AGEs) of aged and cataractous human lenses. The AGEs OP-lysine and argpyrimidine are two UVA absorbing posttranslational modifications that are abundant in the eye lens. The purpose of this study was to outline the changes in these two AGEs due to UVA irradiation. The changes of OP-lysine, OP-phenethylamine (a phenethylamine analogue of OP-lysine), and argpyrimidine due to irradiation with UVA light in the presence or absence of air and ascorbic acid were followed by different spectral methods. Aged human lenses were similarly irradiated in artificial aqueous humor. The amounts of OP-lysine in the irradiated lenses and in the corresponding dark controls were determined by HPLC. Both OP-lysine and argpyrimidine decreased 20% when irradiated with UVA light in the absence of ascorbic acid. Under the same conditions, OP-lysine was bleached 80% in the presence of ascorbic acid during irradiation experiments. In contrast, argpyrimidine UVA light bleaching was not affected by the presence of ascorbic acid. Interestingly the major product of OP-phenethylamine after UVA irradiation in the presence of ascorbic acid was phenethylamine, which indicates that the entire heterocycle of this AGE was cleaved and the initial amino group was restored. Some AGEs in the human eye lens can be transformed by UVA light. PMID- 16037237 TI - The maillard reaction for sunlight protection. AB - During seven months of a clinical trial in spring, summer, and fall, 30 UVA/B/Soret band-photosensitive patients used sequential topical applications of dihydroxyacetone (DHA) followed by naphthoquinone only at bedtime and received excellent photoprotection without a single therapeutic failure or loss of any patient to follow-up. Eighteen of the 30 patients extended the limits of their photoprotection repeatedly over a seven-month period to tolerate without sunburns six to eight hours of midday sunlight under all kinds of occupational and recreational environmental conditions. Previously, the use of 3% DHA topically in earlier studies gave only a sun protection factor (SPF) of 3. In this reanalysis of the original notes of a previous clinical study of the melanoidins produced by DHA followed by naphthoquinone in the keratin layers of the epidermis of minimally pigmented Caucasian photosensitive patients, it is determined that these patients received a minimal UVB photoprotection of SPF 18 or more. This represents at least a sixfold amplification of the UVB photoprotective effect over the use of only dihydroxyacetone in the Maillard reaction. PMID- 16037238 TI - K2P--a novel cross-link from human lens protein. AB - We report here the isolation of a novel acid-labile yellow chromophore from the enzymatic digest of human lens proteins and the identification of its chemical structure by LC-MS and NMR. This new chromophore exhibited a UV absorbance maximum at 343 nm and a molecular mass of 370 Da. One- and two-dimensional NMR analyses elucidated the structure as being 1-(5-amino-5-carboxypentyl)-4-(5-amino 5-carboxypentyl-amino)-3-hydroxy-2, 3-dihydropyridinium, a cross-link between the epsilon-amino groups of two lysine residues and a five-carbon atom ring. We assigned it the trivial name of K2P. Quantitative determinations of K2P in individual normal human lens or cataract lens water-soluble and water-insoluble protein digests revealed a significant enhancement of K2P in the early stage of brunescent cataract lens proteins (type I/II, 613 +/- 362 pmol/mg of water insoluble sonicate supernatant (WISS) protein or 85 +/- 51 pmol/mg of water soluble [WS] protein) when compared with aged normal human lens proteins (261 +/- 93 pmol/mg of WISS protein or 23 +/- 15 pmol/mg of WS protein). Furthermore, a gradual decrease of K2P in the late stages of brunescent cataract lenses with the development of the browning color in the lens argues different coloration mechanisms during the processes of normal aging and cataract development. This new cross-link may serve as a quantitatively significant biomarker for assessing the role of lens protein modifications during aging and in the pathogenesis of cataract. PMID- 16037239 TI - Assessment of protein function following cross-linking by alpha-dicarbonyls. AB - Protein cross-linking via the Maillard reaction with alpha-dicarbonyl compounds has been the subject of intense scrutiny in the literature. We report here a study of the impact of this cross-linking on enzyme function. Protein function following glycation was examined by treating ribonuclease A with methylglyoxal, glyoxal, and diacetyl, which cross-linked the enzyme and impaired its activity. The effects of two reported Maillard reaction inhibitors, aminoguanidine and 3,5 dimethylpyrazole-1-carboxamidine, on the cross-linking reaction were assessed, with a parallel measurement of the effect on enzyme activity. The results demonstrate that preventing protein cross-linking does not necessarily preserve enzyme activity. These results cast doubt on the likely efficacy of some purported antiaging compounds in vivo. PMID- 16037240 TI - Ketosis leads to increased methylglyoxal production on the Atkins diet. AB - In the popular and widely used Atkins diet, the body burns fat as its main fuel. This process produces ketosis and hence increased levels of beta-hydroxybutyrate (BOB) acetoacetate (AcAc) and its by-products acetone and acetol. These products are potential precursors of the glycotoxin methylglyoxal. Since methylglyoxal and its byproducts are recognized as a significant cause of blood vessel and tissue damage, we measured methylglyoxal, acetone, and acetol in subjects on the Atkins diet. We found that by 14-28 days, methylghyoxal levels rose 1.67-fold (P = 0.039) and acetol and acetone levels increased 2.7- and 6.12-fold, respectively (P = 0.012 and 0.028). Samples from subjects with ketosis showed even greater increases in methylglyoxal (2.12-fold), as well as acetol and acetone, which increased 4.19- and 7.9-fold, respectively; while no changes were seen in samples from noncompliant, nonketotic subjects. The increase in methylglyoxal implies that potential tissue and vascular damage can occur on the Atkins diet and should be considered when choosing a weight-loss program. PMID- 16037241 TI - Methylglyoxal, glyoxal, and their detoxification in Alzheimer's disease. AB - The accumulation of advanced glycation end products (AGEs) in brains with Alzheimer's disease (AD) has been implicated in the formation of insoluble deposits such as amyloid plaques and neurofibrillary tangles. AGEs are also known to activate glia, resulting in inflammation and neuronal dysfunction. As reactive intermediates of AGE formation, neurotoxic reactive dicarbonyl compounds such as glyoxal and methylglyoxal have been identified. One of the most effective detoxification systems for methylglyoxal and glyoxal is the glutathione-dependent glyoxalase system, consisting of glyoxalase I and glyoxalase II. In this study, we have determined the methylglyoxal and glyoxal levels in the cerebrospinal fluid of AD patients compared to healthy controls. Methylglyoxal levels in AD patients were twofold higher than in controls, but this difference was not significant due to the large intergroup variations and the small sample size. However, the concentrations of both compounds were five to seven times higher in CSF than in plasma. We also investigated the glyoxalase I level in AD and healthy control brains. The number of glyoxalase I- positive neurons were increased in AD brains compared to controls. Our findings suggest that glyoxalase I is upregulated in AD in a compensatory manner to maintain physiological methylglyoxal and glyoxal levels. PMID- 16037242 TI - Evaluation of glyoxal and methylglyoxal levels in uremic patients under peritoneal dialysis. AB - Advanced glycation end products (AGEs) accumulate in serum and tissues of patients with chronic renal failure, even in the absence of diabetes, and a different clearance of these species has been observed by hemodialysis and peritoneal dialysis (CAPD). Furthermore, it has been shown that not only AGE but also 1,2-dicarbonyl compounds are formed during heat sterilization of glucose based peritoneal dialysis fluids. Therefore, we investigated the level of some AGEs (pentosidine and free pentosidine) and dicarbonyl compounds (glyoxal and methylglyoxal) in end-stage renal disease patients subjected to peritoneal dialysis. Samples (20 from healthy subjects, 16 from uremic patients before and after 12 h of peritoneal dialysis) were analyzed, and the plasma and dialysate levels of glyoxal, methylglyoxal, pentosidine, and free pentosidine were determined. In plasma of uremic patients, mean values of pentosidine showed a small decrease after dialysis and were always higher than those of healthy control subjects. An analogous trend was observed for free pentosidine. In the case of peritoneal dialysate, no pentosidine and free pentosidine were found at time zero, whereas both compounds were detected after 12 h of dialysis. Glyoxal and methylglyoxal mean levels showed a decrease in plasma after dialysis even if their values were always higher than those of healthy control subjects. Surprisingly, an analogous trend was observed also in dialysate. These results might indicate that glyoxal and methylglyoxal already present in the dialysis fluid react with the peritoneal matrix proteins, accounting for the gradual loss of peritoneal membrane function that is often observed in patients subjected to CAPD for a long time. PMID- 16037243 TI - Application of semiquantitative proteomics techniques to the maillard reaction. AB - Proteomic tools-in particular, mass spectrometry (MS)-have advanced significantly in recent years, and the identification of proteins within complex mixtures is now a routine procedure. Quantitative methods of analysis are less well advanced and continue to develop. These include the use of stable isotope ratio approaches, isotopically labeled peptide standards, and nonlabeling methods. This paper summarizes the use of MS as a proteomics tool to identify and semiquantify proteins and their modified forms by using examples of relevance to the Maillard reaction. Finally, some challenges for the future are presented. PMID- 16037244 TI - The carbon module labeling (CAMOLA) technique: a useful tool for identifying transient intermediates in the formation of maillard-type target molecules. AB - Although the Maillard reaction is a well-known source of aroma and taste compounds in processed foods, a systematic correlation of the concentrations of most of the reaction products identified so far with human perception has scarcely been performed. Furthermore, the influence of process parameters on yields and formation mechanisms of key flavor compounds has not been systematically studied. In this short state-of-the-art review, concepts to characterize flavor-active food constituents are briefly discussed, and approaches to elucidate formation mechanisms from labeling experiments and isotopomeric quantitation are highlighted on the basis of results obtained in the author's laboratory. PMID- 16037245 TI - Analysis of protein glycation products by MALDI-TOF/MS. AB - Matrix-assisted laser desorption ionization-mass spectrometry with time-of-flight detection (MALDI-TOF/MS) is a promising tool to analyze advanced glycation end product (AGE)-modified proteins. The combination of soft ionization (MALDI) with time-of-flight mass detection allows analysis of peptides and proteins of a molecular mass up to 300 kDa with minimal sample workup. Because the direct structural analysis of intact AGE proteins is not possible due to the formation of broad and poorly resolved peaks, peptide mapping was introduced into the analysis of AGE proteins by MALDI-TOF/MS, allowing site-specific analysis of defined AGEs. When methylglyoxal-modified lysozyme was subjected to MALDI-TOF/MS peptide mapping, methylimidazolone and argpyrimidine attached to the arginine residue and carboxyethyl (CEL) bound to the lysine were detected on peptide(aa1 7) (KVFGRCE). In contrast, only one methylimidazolone was found on peptide(aa8 35) (LAAAMKRHGLDNYRGYSLGNWVCAAKFE) and peptide(aa120-129) (VQAWIRGCRL), respectively. The analysis of AGE protein, which had been incubated with glucose, revealed the presence of an Amadori product and a carboxymethyl residue (CML) on peptide(aa1-7) and peptide(aa8-35), as well as an imidazolone A on peptide(aa120 129). Furthermore, the early Maillard reaction of lysozyme, which had been glycated by seven different sugars, was monitored by MALDI-TOF/MS peptide mapping. Finally, this approach was successfully applied for site- and product specific relative quantification of AGEs. For example, kinetics of CML and Amadori product formation on peptide(aa1-7), as well as imidazolone A formation on peptide(aa120-129), were determined. PMID- 16037246 TI - Peptide mapping of human serum albumin modified minimally by methylglyoxal in vitro and in vivo. AB - Methylglyoxal is a potent glycating agent and important precursor of advanced glycation end products (AGEs) in physiological systems. Unlike glucose, methylglyoxal is predominantly an arginine-directed glycating agent. Methylglyoxal reacts with proteins to form mainly the arginine-derived hydroimidazolone AGE, Ndelta-(5-hydro-5-methyl-4-imidazolon-2-yl)-ornithine (MG H1), argpyrimidine, the lysine-derived AGEs, N(epsilon)-(1-carboxyethyl)lysine (CEL), and methylglyoxal-derived lysine dimer (MOLD). Sites within proteins susceptible to modification by methylglyoxal have not been identified. Here we show that modification of human serum albumin by methylglyoxal forms mainly hydroimidazolone MG-H1 residues. The location of MG-H1 residues was identified by mass spectrometric peptide mapping. This method identified a hot spot of hydroimidazolone formation at Arg-410, with other minor MG-H1 modifications at Arg-114, Arg-186, Arg-218, and Arg-428. Other extracellular and intracellular proteins are modified by methylglyoxal in physiological systems. Modification of arginine residues by methylglyoxal may be particularly damaging because arginine residues have a high frequency of occurrence in ligand and substrate recognition sites in receptor and enzyme active sites. PMID- 16037247 TI - Advanced glycation end products/peptides: an in vivo investigation. AB - Advanced glycation end products/peptides (AGE/peptides) originate by in vivo enzymatic digestion of nonenzymatically glycated proteins, which are produced by reaction of glucose with primary amino groups present in the protein chain following the Maillard pattern. AGE/peptides are highly reactive species and can interact with tissue and circulating proteins, leading to tissue modification and impaired protein functionality. Serum levels of AGE/peptides are reported to be particularly high in diabetes (in terms of higher production) or in end-stage renal disease (in terms of accumulation). For these reasons, their structural identification is of high interest, giving information on their relationship with the pathological state and allowing the design of possible therapeutic interventions. We report here some preliminary results obtained by liquid chromatography/electrospray ionization/mass spectrometry (LC/ESI/MS) and matrix assisted laser desorption ionization MS (MALDI-MS) investigations carried out on the low-molecular-weight serum peptide fraction from 10 healthy subjects, 10 patients with poorly controlled diabetes, and 10 patients with end-stage nephropathy. PMID- 16037248 TI - A convenient method for preparation of high-purity, Amadori-glycated phosphatidylethanolamine and its prooxidant effect. AB - Phospholipid-linked glycation has been implicated to play an important role in lipid peroxidation associated with several conditions, such as normal aging and diabetes. In this study, we established a convenient method for the preparation of a high-purity, Amadori-glycated phosphatidylethanolamine (Amadori-PE) standard in a methanol system. We evaluated the prooxidant effect of the Amadori-PE standard. When Amadori-PE was incubated in a micellar and liposome system in the presence of metal ion, a remarkable increase of lipid peroxidation was observed in both systems. In addition, the Amadori-PE-induced lipid peroxidation was effectively inhibited by superoxide dismutase, mannitol, catalase, and EDTA. These results indicate that Amadori-PE is a considerable risk factor that may cause membrane lipid peroxidation in the pathogenesis of diabetes and aging. PMID- 16037249 TI - Tandem mass spectrometry analysis of Amadori-glycated phosphatidylethanolamine in human plasma. AB - Amadori-glycated phosphatidylethanolamine (Amadori-PE), a nonenzymatically glycated lipid formed under hyperglycemic conditions, is known as a reliable indicator of lipid glycation in vivo. We have quantified the Amadori-PE concentration in human plasma samples using a reverse-phase liquid chromatography tandem mass spectrometry with neutral loss scan or multiple reaction monitoring. Amounts of Amadori-PE in plasma of diabetic patients (0.15 mol% of PE), diabetic patients with chronic hemodialysis (0.29 mol% of PE), and nondiabetic patients with chronic hemodialysis (0.13 mol% of PE) are higher than that of the control group (0.08 mol% of PE). In addition, the concentration of Amadori-PE was proportional to that of phosphatidylcholine hydroperoxide, a reliable indicator of membrane lipid peroxidation, in human plasma (P < 0.05). These results indicate that plasma Amadori-PE-glycated lipid product formed under hyperglycemic conditions is an inducer of membrane lipid peroxidation, and therefore lipid glycation plays an active part in the development of human disease. PMID- 16037250 TI - Proteomic method for the quantification of methionine sulfoxide. AB - Glycoxidation and lipoxidation reactions contribute to the chemical modification of proteins during the Maillard reaction. Reactive oxygen species, produced during the oxidation of sugars and lipids in these processes, irreversibly oxidize proteins. Methionine is particularly susceptible to oxidation, yielding the oxidation product methionine sulfoxide (MetSO). Here we describe a method for the analysis of MetSO using proteomic techniques. Using these techniques, we measured MetSO formation on the model protein RNase during aerobic incubations with glucose and arachidonate. We also evaluated the susceptibility of MetSO to reduction by NaBH4), a commonly used reductant in the analysis of Maillard reaction products. PMID- 16037251 TI - Simple noninvasive measurement of skin autofluorescence. AB - Accumulation of advanced glycation end products (AGEs) is thought to play a role in the pathogenesis of chronic complications of diabetes mellitus and renal failure. Several studies indicate that AGE accumulation in tissue may reflect the cumulative effect of hyperglycemia and oxidative stress over many years. Simple quantitation of AGE accumulation in tissue could provide a tool for assessing the risk of long-term complications. Because several AGEs exhibit autofluorescence, we developed a noninvasive autofluorescence reader (AFR). Skin autofluorescence measured with the AFR correlates with collagen-linked fluorescence and specific skin AGE levels from skin biopsy samples. Furthermore, skin autofluorescence correlates with long-term glycemic control and renal function, and preliminary results show correlations with the presence of long-term complications in diabetes. The AFR may be useful as a clinical tool for rapid assessment of risk for AGE-related long-term complications in diabetes and in other conditions associated with AGE accumulation. PMID- 16037252 TI - Accumulation of advanced glycation end products, measured as skin autofluorescence, in renal disease. AB - Advanced glycation end products (AGEs) accumulate during renal failure and dialysis. Kidney transplantation is thought to reverse this accumulation by restoring renal function. Using a noninvasive and validated autofluorescence reader, we evaluated AGE levels in 285 transplant recipients (mean age, 52 years; range, 41 to 60 years), 32 dialysis patients (mean age, 56 years; range, 43 to 65 years), and 231 normal control subjects (mean age, 51 years; range, 40 to 65 years). Measurements in transplant recipients were performed for a mean of 73 months (range, 32 to 143 months) after transplantation. Dialysis patients were on dialysis therapy for a mean of 42 months (range, 17 to 107 months). Fluorescence was significantly increased in dialysis patients compared with normal control subjects (2.8 vs. 2.0 arbitrary units [a.u.], P < .0001). Although fluorescence levels were significantly decreased in transplant recipients compared with dialysis patients (2.5 vs. 2.8 a.u., P < .0001), fluorescence in transplant recipients was higher than in controls (2.5 vs. 2.0 a.u., P < .0001). In transplant recipients, fluorescence correlated positively with the duration of dialysis prior to transplantation (R = 0.21, P < .0001), and negatively with creatinine clearance (R = -0.34, P < .0001). No correlation was found between time after transplantation and fluorescence in transplant recipients (R = -0.10, P = .10). Fluorescence in dialysis patients was positively correlated with duration of dialysis (R = 0.36, P = .042). Our results, like those of others, suggest that kidney transplantation does not fully correct increased AGE levels found in dialysis patients. The increased AGE levels in kidney transplant recipients cannot be explained by the differences in renal function alone. The availability of a simple, noninvasive method (AGE-Reader) to measure AGE accumulation may be used to monitor AGE accumulation in a clinical setting as well as in a study setting. PMID- 16037253 TI - Evaluation of the maillard reaction in infant formulas by means of front-face fluorescence. AB - Foods are complex mixtures of macro- and micronutrients, which interact, leading to oxidation, glycation, and hydrolysis upon heating (e.g., sterilization, cooking) and storage. Their nutritional quality and safety are consequently affected, justifying the need for accurate monitoring of the evolution of the food composition during processing and shelf life. Classical chromatographic analysis as well as newly proposed rapid methods based on fluorescence spectrometry analyses were applied on whey powder-based models and commercial samples (in powdered form and ultrahigh temperature [UHT] sterilized), some of which had been previously submitted to protein hydrolysis. These samples were incubated for 48 h at 60 degrees C to mimic accelerated storage. Fluorescence fingerprints addressing modifications in the product composition during processing were recorded and analyzed by chemometric methods. Carboxymethyllysine (Nepsilon-[carboxymethyl]lysine; CML) was measured using an ELISA method. Fluorescence, recorded in a front-face mode on intact samples, is very sensitive to pertinent physicochemical changes induced by heat treatment, formulation (the moisture level in powders, presence of vitamin C and iron), and storage. Similar trends were observed between powders' fluorescence and CML-for example, a strong effect of protein hydrolysis and increasing water content. Addition of vitamin C was associated with an antioxidant effect despite the presence of iron. Good calibration models were obtained for predicting CML from fluorescence spectra both in food models and in commercial samples, although more work is needed to obtain accurate and robust calibration models. Results show the potential of nondestructively applied fluorescence spectrometry for measuring CML in formulas, a rapid, simple, and cost-effective method to monitor formula quality. PMID- 16037254 TI - Interplay between the maillard reaction and lipid peroxidation in biochemical systems. AB - The Maillard reaction and lipid peroxidation are two of the most important chemical reactions that take place in biochemical systems. Both include a whole network of different reactions in which an extraordinarily complex mixture of compounds is produced in very different amounts, with both positive and negative consequences. In addition, both reactions are intimately interrelated, and the products of each reaction influence the other. Furthermore, there are common intermediates and products in both pathways; these products are usually known as advanced glycation end products (AGEs) and advanced lipoxidation end products (ALEs). Moreover, other AGE/ALEs are analogous and participate similarly in both amino acid degradation and amino phospholipid/protein polymerization by identical mechanisms. All these data suggest that the Maillard reaction and lipid peroxidation are so closely interrelated that both reactions should be considered simultaneously to understand the reaction mechanisms, kinetics, and products in the complex mixtures of carbohydrates, lipids, and proteins occurring in biochemical systems. In these systems, lipids and carbohydrates are competing in the chemical modification of amino phospholipids and proteins. Therefore, although there are significant differences between the Maillard reaction and lipid peroxidation, many aspects of both reactions can be better understood if they are included in only one general carbonyl pathway that can be initiated by both lipids and carbohydrates. PMID- 16037255 TI - Isolevuglandins, oxidatively truncated phospholipids, and atherosclerosis. AB - Isolevuglandins (isoLGs) and oxidatively truncated phospholipids are products of lipid peroxidation. Some of these, especially isoLGs and gamma-hydroxyalkenal analogues (e.g., the 5-hydroxy-8-oxo-6-octenoic acid and 9-hydroxy-12-oxo-10 dodecenoic acid esters of 2-lysophosphatidylcholine, HOOA-PC or HODA-PC, respectively) of 4-hydroxy-2(E)-nonenal (HNE), damage proteins by covalent adduction, thereby interfering with their normal functions. These lipid-derived protein modifications may serve as dosimeters of oxidative injury. Elevated plasma levels of isoLG-protein epitopes are associated with atherosclerosis but are independent of total cholesterol, a classical risk factor. Both protein adducts and oxidatively truncated phospholipids (oxPL) can also elicit receptor mediated cellular responses that include endocytosis of oxidized low-density lipoprotein (LDL) and expression of chemokines, which may foster infiltration of monocyte macrophages into the subendothelial space, where they become foam cells through unregulated endocytosis of oxidatively damaged LDL. PMID- 16037256 TI - Role of glucoxidation and lipid oxidation in the development of atherosclerosis. AB - Previous data have indicated that modification of proteins/lipids by glucoxidation and/or lipid oxidation may initiate/propagate the formation of atherosclerotic plaques. Although the biomarker carboxymethyllysine (CML) has been detected in these lesions, the origin of the reactive oxygen species (ROS) leading to its formation and the source of its carbon backbone are unknown. As presented here, the stimulation of cultured monocytes by phorbol-12-myristate-13 acetate (TPA), an activator of protein kinase C that can mimic the effects of high glucose, angiotensin II, and other physiological stimuli, leads to cellular ROS generation and concomitant formation of intracellular CML. Inhibitors of ROS generating cellular systems such as NO synthase, xanthine oxidase, or cytochrome P450 oxidase had no effect on CML formation. Likewise, in cells with inactive NAD(P)H oxidase no reduced CML formation was found. In cells exhibiting a high glycolysis rate, CML formation was unaffected. Because we found rapid CML formation in the presence of unsaturated fatty acids, it appears that lipid oxidation is quantitatively more important. In vivo studies revealed strong intracellular CML staining in areas of histiocytic/monocytic infiltration or proliferation, mostly associated with atheroma formation. Corresponding CML staining patterns were found in healing wounds of different ages, indicating that formation of atherosclerosis is a chronic wound repair associated with a low grade inflammatory reaction. In summary, CML is formed concomitantly with oxidative stress in activated monocytes and can be regarded as a biomarker for a low-grade inflammatory tissue reaction in the atherosclerotic plaque. Its formation via lipid oxidation may be involved in the development of atherosclerosis. PMID- 16037257 TI - Is atherosclerosis a multifactorial disease or is it induced by a sequence of lipid peroxidation reactions? AB - The delivery of not only free cholesterol but also cholesterol esters to cells by low-density lipoprotein (LDL) has hitherto been unstudied. Minor compounds present in mammalian-derived food include cholesterol linoleate and arachidonate. Evidence is presented that these esters are directly incorporated into VLDL and are responsible for the deleterious effects of atherosclerosis. Cholesterol esterified with these polyunsaturated fatty acids (PUFAs) is readily oxidized at the PUFA residue during storage and heating. Apparently, the liver is unable to distinguish between nonoxidized and oxidized cholesterol PUFA esters and also incorporates the latter into VLDL, which is transformed to LDL. When this LDL is transferred to endothelial cells, the toxic products are liberated and induce cell damage. Cell damage is combined with structural changes that influence neighboring cells and cause an influx of Ca2+ ions and activation of phospholipases and lipoxygenases, resulting in production of lipid hydroperoxides (LOOHs). When the level of free PUFAs generated by phospholipases exceeds a certain limit, lipoxygenases commit suicide, causing liberation of iron ions. The latter react with LOOHs and thus induce a switch from enzymatic to nonenzymatic generation of lipid peroxidation (LPO) products. Although the LOO. radicals produced in enzymatic reactions are deactivated within the enzyme complex, LOO. radicals generated in nonenzymatic reactions are able to attack any biological compound, inducing severe damage. Apparently, iron ions and LOOH molecules at the surface of injured cells transfer the nonenzymatic LPO reactions to the phospholipid layer of bypassing lipoproteins, thus explaining why inflammatory diseases, such as diabetes, are combined with atherogenesis. PMID- 16037258 TI - The immunogenicity of modified lipoproteins. AB - The immunogenicity of modified low-density lipoprotein (mLDL) has been demonstrated both in laboratory animals and humans. Circulating human mLDL antibodies, purified by affinity chromatography, are predominantly of the IgG isotype, subclasses 1 and 3. The purified antibodies react with malondialdehyde lysine and carboxymethyl-lysine epitopes, but also recognize minimally modified forms of LDL that do not contain significant amounts of those two epitopes. The quantitative assays of mLDL and mLDL antibodies in serum samples by enzymoimmunoassay (EIA) are unreliable owing to the interference of preformed circulating immune complexes (CICs). Isolation of CICs by precipitation with low concentrations of polyethylene glycol followed by analysis of antigens and antibodies contained in the precipitates is a technically complex approach, but one that yields valuable data. With this approach we have confirmed that the IgG antibodies involved in IC formation belong to the proinflammatory IgG1 and IgG3 isotypes, have a higher avidity than those that remain unbound in the supernatant after CIC precipitation, and are of higher avidity in diabetic patients with macroalbuminuria than in those with normal albuminuria. We have also developed capture assays for different forms of mLDL. These assays have shown a significant enrichment in mLDL of the precipitated ICs. The enrichment is also more pronounced in the CICs obtained from diabetic patients with macroalbuminuria. In conclusion, isolation and characterization of LDL-ICs appears to yield information of significant value that is not derived from other approaches to measure LDL modifications and their corresponding antibodies in humans. PMID- 16037259 TI - Glycation does not alter LDL-induced secretion of tissue plasminogen activator and plasminogen activator inhibitor-1 from human aortic endothelial cells. AB - Diabetes may induce both quantitative and qualitative changes in lipoproteins, especially low-density lipoprotein (LDL). Effects of LDL glycation on endothelial cell secretion of tissue plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1) have not been fully elucidated. Human aortic endothelial cell (HAEC) tPA and PAI-1 production were determined after incubation with LDL (50 to 500 microg/mL protein, 24 h) from three sources: (1) nondiabetic LDL (N-LDL) modified in vitro to form six preparations: native, nonmodified (N); glycated (G); minimally oxidized (MO); minimally oxidized and glycated (MOG); heavily oxidized (HO); and heavily oxidized and glycated (HOG); (2) in vivo glycated and relatively nonglycated LDL subfractions from type 1 diabetic patients; (3) LDL from type 1 diabetic patients and matched controls, which was subfractionated using density gradient ultracentrifugation. In experiments using LDL modified in vitro, the rate of tPA release by HAECs incubated with N-LDL (83 +/- 4 ng/mg cell protein/24 h) did not differ significantly from those incubated with G-LDL (73 +/ 7), MO-LDL (74 +/- 13), or MOG-LDL (66 +/- 15) and was not influenced by LDL concentration. The rate of PAI-1 release was similar in HAECs incubated with N LDL (5.7 +/- 0.6 mug/mg cell protein/24 h), G-LDL (5.7 +/- 0.7), MO-LDL (5.5 +/- 0.8), or MOG-LDL (5.7 +/- 0.9) and was not influenced by LDL concentration. In contrast, tPA release was significantly decreased in cells incubated with LDL (10 microg/mL) modified extensively by oxidation, and averaged 45.2 +/- 5.0 and 43.7 +/- 9.9 ng/mg/24 h for HO-LDL and HOG-LDL, respectively, and was further decreased with increasing concentrations of the heavily oxidized LDL preparations. PAI-1 release was not significantly decreased relative to N-LDL in cells incubated with low concentrations (5 to 50 microg/mL) of HO-LDL and HOG LDL, but was decreased to 3.2 +/- 0.5 and 3.1 +/- 0.7 microg/mg/24 h for HO-LDL and HOG-LDL at 200 microg/mL, respectively. Results using in vivo glycated versus nonglycated LDL showed that tPA and PAI-1 release did not differ between subfractions. Release of tPA averaged 5.11 +/- 0.6 and 5.12 +/- 0.7 ng/mg/24 h, whereas release of PAI-1 averaged 666 +/- 27 ng/mg/24 h and 705 +/- 30 ng/mg/24 h for nonglycated and glycated LDL subfractions, respectively. Using LDL of different density subclasses, tPA and PAI-1 release in response to LDL from diabetic patients compared with control subjects did not differ when HAECs were incubated with LDLs of increasing density isolated from each subject pair. We conclude that oxidation of LDL, but not glycation, may contribute to the altered fibrinolysis observed in diabetes. PMID- 16037260 TI - Effects of modified low-density lipoproteins on human retinal pericyte survival. AB - According to a current paradigm cardiovascular diseases can be initiated by exposure of vascular cells to qualitatively modified low-density lipoproteins (LDL). Capillary leakage, an early feature of diabetic retinopathy, results in the exposure of retinal pericytes to modified LDL, including glycated (G-LDL) and heavily oxidized glycated LDL (HOG-LDL). We demonstrate here that modified LDL inhibits the proliferation and survival of cultured human retinal pericytes. Modified LDL also induced DNA fragmentation in bovine retinal pericytes. Overall, HOG-LDL produced a significantly higher extent of cytotoxicity and apoptosis in retinal pericytes. These results indicate that exposure of pericytes to HOG-LDL could be implicated in the development of diabetic retinopathy. PMID- 16037261 TI - Acrolein modifies apolipoprotein A-I in the human artery wall. AB - Carbonyl stress is implicated in accelerated vascular disease, but little is known about the factors that control the reactions of carbonyls with proteins. Acrolein is a reactive carbonyl generated by the oxidation of lipids and amino acids. It also forms during cigarette smoking. We therefore investigated the possibility that acrolein might react with apolipoprotein A-I (apoA-I), the major protein of high-density lipoprotein (HDL), which plays a critical role in mobilizing cholesterol from artery wall macrophages. Tandem mass spectrometric analysis demonstrated that lysine residues were the only amino acids in apoA-I that were modified by acrolein. Immunohistochemical studies with a monoclonal antibody revealed that acrolein adducts colocalized with apoA-I in human atherosclerotic lesions. Moreover, the ability of apoA-I to remove cholesterol from cultured cells was impaired after exposure to acrolein, suggesting that the carbonyl might interfere with apoA-I's normal function of promoting cholesterol efflux from artery wall cells. Our observations suggest that acrolein may interfere with normal HDL cholesterol transport by modifying apoA-I. This structural damage might play a critical role in atherogenesis by impairing cholesterol removal from artery wall cells. PMID- 16037262 TI - Lipid-derived modifications of plasma proteins in experimental and human diabetes. AB - Plasma from two diabetic rat models and human diabetic patients was analyzed to investigate the hypothesis that enhanced oxidative stress in diabetes promotes lipid-derived protein modification. We evaluated the nonenzymatic modification of plasma protein by oxidized phospholipids, including measurement of protein-bound pentanedioate, nonanedioate, and hexanoate, all derived from oxidation of phospholipid polyunsaturated fatty acids. Generally pentanedioate was higher in diabetic compared with nondiabetic control groups, and nonanedioate was also higher in the diabetic rat models. We conclude that diabetes is associated with higher levels of phospholipid-derived protein modification in both animal models and human diabetes. Their role in the development of diabetes vascular complications warrants further research. PMID- 16037263 TI - Angiogenic potency of Amadori-glycated phosphatidylethanolamine. AB - Glycation has been thought to participate in diabetic vascular diseases. However, there are no reports about the effects of lipid glycation on endothelial dysfunction. In this study, we have evaluated whether Amadori-glycated phosphatidylethanolamine (Amadori-PE), a lipid-linked glycation compound, affected proliferation, migration, and tube formation of cultured human umbilical vein endothelial cells. These three factors involved in angiogenesis were significantly stimulated by Amadori-PE at a low concentration of less than 5 microM. Furthermore, Amadori-PE also stimulated the secretion of matrix metalloproteinase-2 (MMP-2), a pivotal enzyme in the initial step of angiogenesis. Our results indicated for the first time that Amadori-PE would elicit vascular disease through angiogenic potency on endothelial cells, thereby playing an active part in the development and progression of diabetic microangiopathy. PMID- 16037264 TI - Covalent binding of acetone to aminophospholipids in vitro and in vivo. AB - We have determined the ions characteristic of acetone adducts of reference aminophospholipids and have used them as markers for identification of acetone adducts of aminophospholipids in commercial lecithin, acetone extracts of tissue lipids, and in plasma and red blood cells of diabetic subjects. The acetonation products were determined by normal-phase high-performance liquid chromatography (HPLC) with on-line electrospray-mass spectrometry, and electrospray/collision induced dissociation in the negative ion mode. The major acetone complexes of PtdEtn and PtdSer were identified as the diacetone derivatives [PtdEtn+116-H2O]- and [PtdSer+116-H2O]-, respectively, although ions corresponding to monoacetone [PtdEtn+58-H2O]- and doubly dehydrated diacetone adducts [PtdSer+116-2 x 18]- were also observed. Upon increase of the capillary exit voltage (CapEx) from -160 to -300 V, new ions appeared with the original retention time but with 58 masses (one acetone molecule) lower than the mass of the parent compounds, along with fragment ions corresponding to lysoGPE+40 and free fatty acids. Scanning of chloroform/methanol extracts of red blood cell lipids of two of five diabetic subjects examined yielded elevated levels (in relation to nondiabetic subjects) for ions corresponding to the diacetone adducts [M+98]- of the major molecular species of PtdEtn and PtdSer. Because of possible overlap with major molecular species of PtdIns, the identification of the acetonated PtdSer in diabetic blood requires further confirmation. PMID- 16037265 TI - Protective role of antioxidative food factors in oxidative stress caused by hyperglycemia. AB - Hyperglycemia causes the autoxidation of glucose, glycation of proteins, and the activation of polyol metabolism. These changes accelerate generation of reactive oxygen species (ROS) and increases in oxidative chemical modification of lipids, DNA, and proteins in various tissues. Oxidative stress may play an important role in the development of complications in diabetes such as lens cataracts, nephropathy, and neuropathy. Glycation reactions, especially Maillard reactions, occur in vivo as well as in vitro and are associated with the chronic complications of diabetes mellitus and aging and age-related diseases by increases in oxidative chemical modification of lipids, DNA, and proteins. In particular, long-lived proteins such as lens crystallines, collagens, and hemoglobin may react with reducing sugars to form advanced glycation end products (AGEs). Recently, we found a novel type of AGE, named MRX, and we found that MRX is a good biomarker for detecting oxidative stress produced during Maillard reaction. We also examined in detail the role of lipid peroxidation reaction in hyperglycemia and found that hexanoyl modification formed by the reaction of oxidized lipids and proteins must be important for oxidative stress. Detailed analyses of the formation mechanism of hexanoyl lysine (HEL) moiety in proteins were conducted, and excretion of HEL into urine was quantified by using LC/MS/MS. Macrophages and neutrophils play an important role in oxidative stress during hyperglycemia, and we determined that oxidatively modified tyrosines are a good biomarker for formation of oxidative stress at an early stage. Immunochemical analyses by application of monoclonal antibodies specific to lipid hydroperoxide modified proteins produced by polyunsaturated fatty acids including docosahexaenoic acid (DHA) in oxidative stress caused by hyperglycemia were conducted, and the relationship between glycation and lipid peroxidation reactions both by chemical and immunochemical approaches are discussed. Recently, we put much more focus on dietary antioxidants for prevention of diabetic complications. Curcuminoids, the main yellow pigments in Curcuma longa (turmeric), have been used widely and for a long time in the treatment of sprain and inflammation in indigenous medicine. Curcumin is the main component of turmeric, and two minor components are also present as the curcuminoids. Curcuminoids possess antioxidant activity. Protective effects of curcumin (U1) and one of its major metabolites, tetrahydrocurcumin (THU1), have been examined for development of diabetic cataract in 25% galactose-fed SD rats. Through detailed examination of protective mechanisms of THU1, it was found that THU1 showed that scavenger ROS not only formed during hyperglycemia, but also induced antioxidative enzymes including detoxification enzymes such as glutathine S transferase. THU1 also showed significant increase of glutathione concentration in the cultured rat lens. Glutathione (gamma-glutamylcysteinyl glycine [GSH]) is thought to be an important factor in cellular function and defense against oxidative stress, and we found that dietary GSH suppresses oxidative stress in vivo in prevention of diabetic complications such as diabetic nephropathy and neuropathy. PMID- 16037266 TI - Advanced glycation in health and disease: role of the modern environment. AB - It is believed that intracellular and extracellular advanced glycation (AGEs) or lipoxidation end products (ALEs), together with dysregulated glucose and lipid metabolism, are important contributors to oxidant or carbonyl stress, enhanced cellular redox-sensitive transcription factor activity, and impaired innate immune defense, causing over time inappropriate inflammatory responses. However, neither the magnitude nor the persistent nature of this increased prooxidant state are completely understood. A significant correlation has been found between ingested and circulating AGEs in humans in recent years. Based on animal studies, the injurious impact of diet-derived AGEs to vascular and kidney tissues is estimated to rival or even exceed that caused by hyperglycemia or hyperlipidemia. Consistent with this view, dietary AGE restriction has been associated with suppression of several immune defects, insulin resistance, and diabetic complications, whether genetically or diet induced, despite persistent diabetes. These findings are in support of clinical evidence from subjects with diabetes or vascular or kidney disease. Most recently, evidence from animal studies points to AGE restriction as an effective means for extending median life span, similar to that previously shown by marked caloric restriction. We conclude that excessive AGE consumption, in the current dietary/social structure, represents an independent factor for inappropriate oxidant stress responses, which may promote the premature expression of complex diseases associated with adult life, such as diabetes and cardiovascular disease. PMID- 16037267 TI - Diet-derived advanced glycation end products are major contributors to the body's AGE pool and induce inflammation in healthy subjects. AB - Advanced glycation end products (AGEs) are a heterogeneous group of compounds that form continuously in the body. Their rate of endogenous formation is markedly increased in diabetes mellitus, a condition in which AGEs play a major pathological role. It is also known, however, that AGEs form during the cooking of foods, primarily as the result of the application of heat. This review focuses on the generation of AGEs during the cooking of food, the gastrointestinal absorption of these compounds, and their biological effects in vitro and in vivo. We also present preliminary evidence of a direct association between dietary AGE intake and markers of systemic inflammation such as C-reactive protein in a large group of healthy subjects. Together with previous evidence from diabetics and renal failure patients, these data suggest that dietary AGEs may play an important role in the causation of chronic diseases associated with underlying inflammation. PMID- 16037268 TI - Are food advanced glycation end products toxic in biological systems? AB - Model food advanced glycation end products (AGEs) were prepared as glycated casein (GC) and glycated soy protein (GS) by the reaction of casein or soy protein with glucose at 50 degrees C, relative humidity 75% for seven days in a powder state. These browned proteins were used as materials for animal experiments. A mixture of 20% glycated proteins (GC:GS = 1:1) diet was fed to streptozotocin (STZ)-diabetic rats for 11 weeks. The results showed that: (1) fructoselysine was observed in the hepatic portal veins, arteries, and femoral veins of rats fed with glycated proteins after 2 h of feeding; (2) blood sugar of glycated protein-fed rats was lower than that of diabetic rats fed with intact protein, while HbA1C in blood and glucose in urine of both groups were similar; (3) lipid peroxidation status in serum, liver, and kidney of both groups was similar; (4) superoxide dismutase (SOD) and glutathione-S-transferase (GST) enzymatic activity in serum and liver of both groups were also similar; (5) there were no differences in degree of cataract formation and concentration of glucose, fructose, sorbitol, and lipid peroxide in the lenses of both groups. From the above results, it can be estimated that food AGEs are not toxic in biological systems, and reactive oxygen species increase in diabetic rats is not caused by glycated proteins but by other pathways. PMID- 16037269 TI - Studies on absorption and elimination of dietary maillard reaction products. AB - A nine-day dietary study involving 18 healthy volunteers was performed in order to investigate the influence of nutrition on the urinary excretion of the Maillard reaction products (MRPs) fructoselysine, pyrraline, and pentosidine. From day two through day eight, most types of Maillard product-containing food had to be avoided. On day five, participants were divided into four groups, three of them receiving a test meal (pretzel sticks, brewed coffee, or custard) containing defined amounts of MRPs. The fourth group served as a control. Urine samples taken over a 24-h period were analyzed for MRPs using chromatographic means. As a result of the MRP-free diet, urinary excretion of free pyrraline and fructoselysine, which was calculated from furosine analysis, were lowered about 90%. Excretion of pentosidine decreased about 40%. Consumption of pretzel sticks and coffee on day five resulted in increased amounts of pyrraline and pentosidine in urine samples on days five to seven. Related to the supplied amounts of pyrraline, about 50% were recovered in the urine samples after ingestion of the pretzel sticks. For pentosidine, 60% of the ingested free derivative from coffee brew and 2% of the peptide-bound amino acid ingested with the bakery product were recovered in the urine samples, indicating a better bioavailability for free pentosidine compared to the protein-bound form. For peptide-bound Amadori products, no influence on the excretion was observed after ingestion of the test foods, indicating degradation in the intestine or plasma to yet-unknown metabolites. In conclusion, differences concerning the excretion rate of individual MRPs point to individual resorption and metabolic pathways. These results are of importance for the discussion of a possible (patho)physiological role of dietary advanced glycation end products (AGEs). PMID- 16037270 TI - Renal effects of oral maillard reaction product load in the form of bread crusts in healthy and subtotally nephrectomized rats. AB - The biological consequences of chronic consumption of Maillard reaction products (MRPs) on renal function in health and renal disease are still incompletely understood. We investigated the metabolic and renal effects of a diet with varying MRP content in healthy and subtotally nephrectomized rats. Male Wistar rats were subjected to sham operation (control, C, n = 12), or to 5/6 nephrectomy (5/6NX, n = 12). Both groups were randomized into subgroups and pair-fed with either a MRP-poor or -rich diet for six weeks. The diet was prepared by replacing 5% or 25% of wheat starch by bread crust (BC). In spite of pair-feeding, the rats on the 25% BC diet gained more body weight (C: 183 +/- 6 g; C + 5% BC: 197 +/- 7 g; C + 25% BC: 229 +/- 6 g [P < 0.05]; 5/6NX: 165 +/- 10 g; 5/6NX + 5% BC: 202 +/ 3 g; 5/6NX + 25% BC: 209 +/- 8 g [P < 0.05]) and had a higher organ weight (heart, liver, lung, kidney/remnant kidney). Bread crust-enriched diet induced proteinuria (C: 15 +/- 5 mg/24 h; C + 5% BC: 19 +/- 4; C + 25% BC: 26 +/- 3 [P < 0.05]; 5/6NX: 30 +/- 7 mg/24 h; 5/6NX + 5% BC: 47 +/- 9; 5/6NX + 25% BC: 87 +/- 19 [P < 0.01]) and a rise in urinary transforming growth factor beta(1) excretion (C: 0.4 +/- 0.1 ng/24 h; C + 5% BC: 0.6 +/- 0.1; C + 25% BC: 1.2 +/- 0.3; 5/6NX: 0.5 +/- 0.1 ng/24 h; 5/6NX + 5% BC: 0.9 +/- 0.1; 5/6NX + 25% BC: 1.6 +/- 0.2 [P < 0.01]). Plasma creatinine or creatinine clearance were not affected significantly. In conclusion, our data suggests that long-term consumption of a diet rich in MRPs may lead to damage of the kidneys. PMID- 16037271 TI - Dietary bread crust advanced glycation end products bind to the receptor for AGEs in HEK-293 kidney cells but are rapidly excreted after oral administration to healthy and subtotally nephrectomized rats. AB - In renal HEK-293 cells, the dietary Maillard reaction compounds casein-linked Nepsilon-carboxymethyllysine (CML), CML, bread crust (BC), and pronyl-glycine (a key compound formed in association with the process-induced heat impact applied to bread dough) all showed activation of p38-MAP kinase. Expression of the C terminus truncated receptor for advanced glycation end products (RAGE) resulted in a reduction of HEK-293-MAP kinase activation. As these findings suggested a RAGE-mediated activating effect of CML, BC, and pronyl-glycine on kidney cellular signal transduction pathways, an in vivo study was performed. Male Wistar rats were subjected to a sham operation (CTRL, n = 20) or to 5/6 nephrectomy (NX, n = 20). Both groups were randomized into two subgroups and fed 20 g of a diet containing either 25% by weight BC or wheat starch (WS). GC-MS analyses of CML, carboxyethyllysine (CEL), and pentosidine revealed increased levels of CML and CEL in the liver but decreased levels of CML in the kidneys of CTRL and NX rats fed the BC diet compared to those on the WS diet. However, urinary levels of CML were also elevated in the CTRL and NX rats on the BC diet, pointing to enhanced excretion of AGEs after BC administration. Although renal insufficiency in the NX rats was reflected by proteinuria, the renal handling of CML and, presumably, other AGEs was not impaired. PMID- 16037272 TI - Biological and chemical assessment of antioxidant activity of sugar-lysine model maillard reaction products. AB - The antioxidant activity of Maillard reaction products (MRPs) is often associated with increased stability and shelf life of food systems vulnerable to oxidation reactions. In this study, nondialyzed, high-molecular weight (HMW = >3500 Da) MRPs were recovered from three model sugar-lysine (glucose-lysine, Glc-Lys; fructose-lysine, Fru-Lys; and ribose-lysine, Rib-Lys) reactions, heated at 121 degrees C for one hour. Samples were characterized by UV and fluorescence spectra and assessed for antioxidant activity using both standard chemical methods (1,1 diphenyl-2-picryl-hydrazyl [DPPH] and oxygen radical absorbing capacity [ORAC]). In addition, biochemical (e.g., cell culture for intracellular oxidation in RAW264.7 cells and protection against metal ion-induced cytotoxicity in C3H/10T1/2 mouse embryo fibroblast cells) assays were used. Patterns of change for fluorescence and multiple colorimetric parameters corresponded to the recovery yield of HMW MRPs and indicated that Rib was more (P < 0.05) reactive than Glc, which in turn was greater (P < 0.05) than Fru. These characteristics of rate of browning did not parallel the significant (P < 0.05) antioxidant activity noted for different sugar-derived HMW MRPs to scavenge DPPH radical, or exhibit total antioxidant activity using the ORAC (e.g., 800-1000 micromol Trolox/gm MRP) method. Antioxidant activity of Glc-, Fru-, and Rib-Lys HMW-MRPs (50 microg/mL) produced protection (P < 0.05) against both H2O2- and AAPH-induced intracellular oxidation reactions in cultured RAW 264.7 cells. Metal chelating activity of all three sugar-derived HMW MRPs (0.01% w/v) was attributed to similar protection (P < 0.05) against Fe2+ and Cu2+-induced cytotoxicity in cultured mouse embryonic fibroblasts. The reducing activity of all three HMW-MRPs indicated the potential for prooxidant activity that could explain enhanced cytotoxicity of Fe3+ in cultured cells. PMID- 16037273 TI - Prevention of oxidative stress by adenoviral overexpression of glutathione related enzymes in pancreatic islets. AB - Chronic exposure to supraphysiologic glucose concentrations causes functional damage to cells and tissues, a process known as glucose toxicity. Recent research indicates that one important mechanism for glucose toxicity is oxidative stress. Glucose has been shown to form reactive oxygen species through several metabolic pathways. The pancreatic islet is distinguished by its relatively low antioxidant enzyme content and activity, which render it especially susceptible to oxidative stress. Adenoviral overexpression of glutathione peroxidase as well as gamma glutamylcysteine ligase have been shown to protect the islet against oxidative stress. Antioxidants have been shown to brake the worsening of diabetes by improving beta cell function in animal models. These observations suggest that enhancing antioxidant defense mechanisms in pancreatic islets may be a valuable pharmacologic approach to managing diabetes. PMID- 16037274 TI - A common pathway for intracellular reactive oxygen species production by glycoxidative and nitroxidative stress in vascular endothelial cells and smooth muscle cells. AB - A large body of evidence suggests that carbonyl compounds induce intracellular signaling by increasing oxidative stress in the cell; however, the mechanisms involved have not been fully described. The focus of our research is on the pathway in which antioxidative enzymes are modified and inactivated by carbonyl compounds, resulting in the accumulation of active oxygen species in the cell. A common pathway appears to exist for cellular signaling evoked by nitroxidative stress. It could be concluded that some glycoxidative stress and nitroxidative stress cause intracellular signaling via similar mechanisms. The elucidation of the pathway for extracellular stress-induced reactive oxygen species (ROS) production would be important for our understanding of the role of ROS as signaling molecules. PMID- 16037275 TI - An in vitro approach to the chronological aging of skin by glycation of the collagen: the biological effect of glycation on the reconstructed skin model. AB - Glycation is a slow, nonenzymatic reaction that takes place between free amino groups in proteins primarily from lysine and a reducing sugar such as glucose or ribose. In skin, this reaction creates new residues or formations of cross-links (advanced glycation end products, AGEs) in the extracellular matrix of the dermis. The formation of these bridges between dermal molecules is supposed to be responsible for loss of elasticity or other properties of the dermis observed during aging. Glycation may therefore play an important role in chronologic aging. In order to examine this hypothesis, we have developed a reconstructed skin model made of a modified dermal compartment that is a fibroblast-contracted collagen lattice prepared with preglycated collagen. The presence of AGEs (glycoxidation products) in the skin equivalents was evidenced using specific antibodies against carboxymethyllysine (CML). Several changes were observed after collagen glycation: (1) fibroblast shape and distribution (vimentin staining) were modified; (2) extracellular matrix molecules and the dermal-epidermal junction zone seemed to be enhanced (procollagen I and III, collagen IV and VII stainings); (3) stainings for beta1 and alpha6 integrins were also increased in the epidermal cell layer; and (4) collagenase activity was increased. To verify the biological effect of glycation, we used the well-known glycation inhibitor aminoguanidine. After aminoguanidine treatment, we found a low CML amount and decreased distribution of markers previously overexpressed in glycated skin constructs. These in vitro findings were at least in part related to aging in vivo and demonstrate an actual effect of glycation in skin aging. PMID- 16037276 TI - Cross-linking of the extracellular matrix by the maillard reaction in aging and diabetes: an update on "a puzzle nearing resolution". AB - The aging extracellular matrix is characterized by an age-related increase in insolubilization, yellowing, and stiffening, all of which can be mimicked by the Maillard reaction in vitro. These phenomena are accelerated in metabolic diseases such as diabetes and end-stage renal disease, which have in common with physiological aging the accumulation of various glycation products and cross links. Eight years ago we concluded that the evidence favored oxidative cross linking in experimental diabetes [Monnier, V.M. et al. 1996. The mechanism of collagen cross-linking in diabetes: a puzzle nearing completion. Diabetes 45(Suppl. 3): 67-72] and proposed a major role for a putative non-UV active cross link derived from glucose. Below, we provide an update of the field that leads to the conclusion that, while oxidation might be important for Maillard reaction mediated cross-linking via Strecker degradation and allysine formation, the single most important collagen cross-link known to date in diabetes and aging is glucosepane, a lysyl-arginine cross-link that forms under nonoxidative conditions. PMID- 16037277 TI - Oxidative stress and neurodegeneration. AB - Oxidative stress is a well-studied early response in chronic neurodegenerative diseases, including Alzheimer's disease, where neuronal loss can exceed 90% in the vulnerable neuronal population. Oxidative stress affects all classes of macromolecules (sugar, lipids, proteins, and DNA), leading inevitably to neuronal dysfunction. We observed that Nepsilon-(carboxymethyl)lysine (CML), the predominant advanced glycation end product that accumulates in vivo, along with its glycation-specific precursor hexitol-lysine, are increased in neurons from cases of Alzheimer's disease, especially those containing intracellular neurofibrillary pathology. The increase in hexitol-lysine and CML can result from either lipid peroxidation or advanced glycation, whereas hexitol-lysine is solely a product of glycation, suggesting that two distinct oxidative processes act in concert in the neuropathology of the disease. Furthermore, using olfactory neurons as an experimental model, we observed an increase in glycation products in neurons derived from Alzheimer's disease patients. Our findings support the idea that aldehyde-mediated modifications, in concert with oxyradical-mediated modifications, are critical early pathogenic factors in Alzheimer's disease. PMID- 16037278 TI - Receptor for advanced glycation end products and its ligands: a journey from the complications of diabetes to its pathogenesis. AB - Many studies have suggested that the expression of RAGE (receptor for advanced glycation end products) is upregulated in human tissues susceptible to the long term complications of diabetes. From the kidneys to the macrovessels of the aorta, RAGE expression is upregulated in a diverse array of cell types, from glomerular epithelial cells (podocytes) to endothelial cells, vascular smooth muscle cells, and inflammatory mononuclear phagocytes and lymphocytes. Although RAGE was first described as a receptor for advanced glycation end products (AGEs), the key finding that RAGE was also a signaling receptor for proinflammatory S100/calgranulins and amphoterin, led to the premise that even in euglycemia, ligand-RAGE interaction propagated inflammatory mechanisms linked to chronic cellular perturbation and tissue injury. Indeed, such considerations suggested that RAGE might even participate in the pathogenesis of type 1 diabetes. Our studies have shown that pharmacological and/or genetic deletion/mutation of the receptor attenuates the development of hyperglycemia in NOD mice; in mice with myriad complications of diabetes, interruption of ligand RAGE interaction prevents or delays the chronic complications of the disease in both macro- and microvessel structures. Taken together, these findings suggest that RAGE is "at the right place and time" to contribute to the pathogenesis of diabetes and it complications. Studies are in progress to test the premise that antagonism of this interaction is a logical strategy for the prevention and treatment of diabetes. PMID- 16037279 TI - Receptor for advanced glycation end products is a promising target of diabetic nephropathy. AB - Advanced glycation end products (AGEs) and the receptor for AGE (RAGE) interactions have been implicated in the development of diabetic vascular complications, which cause various disabilities and shortened life expectancy, and reduced quality of life in patients with diabetes. Diabetes-induced RAGE overexpressing transgenic mice exhibited the exacerbation of the indices of nephropathy, and this was prevented by the inhibition of AGE formation. We also created RAGE-deficient mice by homologous recombination. They showed marked amelioration of diabetic nephropathy as compared with wild-type mice. Through an analysis of vascular polysomal poly(A)+ RNA, we identified a novel splice variant coding for a soluble RAGE protein and named it endogenous secretory RAGE (esRAGE). esRAGE was able to protect AGE-induced vascular cell injuries as a decoy receptor and was actually detected in human circulation. We conclude that RAGE plays an active role in the development of diabetic vascular complications, especially nephropathy, and is a promising target for overcoming this disease. The esRAGE, an endogenous decoy receptor, may be related to individual variations in resistance to the development of diabetic vascular complications. PMID- 16037280 TI - Glycation products as markers and predictors of the progression of diabetic complications. AB - The structure of a growing number of glycation and advanced glycation end products has been elucidated. Measuring these products can be used to assess cumulative glycemic and glycoxidative damage in diabetes and other chronic conditions. The predictive power of a given glycation product can be tested in large prospective studies that evaluate the risk of developing diabetic micro- and macrovascular disease over years following the quantitative determination of that marker. This article provides a comprehensive review of the field, comparing the merits of each marker, whether in skin, serum, or other tissue. Several conclusions are drawn, one of which identifies skin glycation products as powerful predictors of the risk of developing diabetic complications. PMID- 16037281 TI - The maillard reaction in eye diseases. AB - Diabetes and age-related eye disorders remain leading causes of blindness worldwide. While defined pathogenic mechanisms for many of these diseases remain elusive, there is increasing evidence that products of the Maillard reaction may play an important role in their etiology. Advanced glycation end products (AGEs) form though a range of pathways within Maillard chemistry, and there is evidence to suggest that these adducts accumulate in the intracellular and/or extracellular environment of ocular structures. This review evaluates the ever growing literature on AGEs in biological systems and draws relevant links to diseases such as diabetic retinopathy, age-related macular degeneration, and cataract formation. It also outlines recent pharmaceutical strategies to inhibit Maillard reaction products and provides links to how these may serve to limit ocular cell dysfunction. PMID- 16037282 TI - Role of advanced glycation end products and their receptors in development of diabetic neuropathy. AB - Diabetic neuropathy is a life-threatening complication involving both peripheral and autonomic nerves. The hyperglycemia-induced polyol pathway as well as enhanced oxidative stress are among the factors implicated in the pathogenesis of diabetic neuropathy. Their effects are possibly exerted by direct nerve tissue damage or mediated by endothelial injury or vascular dysfunction. Formation of advanced glycation end product (AGE) is another important candidate for the cause of peripheral neuropathy. Indeed, the levels of AGEs were increased in the serum and also in the peripheral nerves obtained from diabetic patients. Structural and functional proteins of those nerves are also glycated, resulting in impaired nerve function and characteristic pathologic alterations. In addition, interaction between AGEs and their receptors induce biological effects on the target tissues for diabetic complications. In the peripheral nerve, the receptor for AGE (RAGE) is expressed in endothelial and Schwann cells. It is thus anticipated that interactions between AGEs and RAGE facilitate endoneural vascular dysfunction, leading to microangiopathy in the peripheral nerve. The roles of these mechanisms, in particular on the molecular mechanisms of AGE-RAGE interactions in the development of diabetic neuropathy are largely still speculative and yet to be explored. PMID- 16037283 TI - Synergistic contributions of carbonyl stress and megsin in diabetic nephropathy. AB - Diabetic nephropathy is the most common cause of end-stage renal failure. The primary glomerular changes in diabetic nephropathy are diffuse and nodular glomerulosclerosis, manifested by an increase in mesangial matrix. Research has demonstrated that advanced glycation end products (AGEs), oxidative stress, and carbonyl stress might play a crucial role in the pathogenesis of diabetic nephropathy via multiple mechanisms. AGEs augment extracellular matrix synthesis, contribute to the release of proinflammatory cytokines and expression of growth factors and adhesion molecules, and interact with the renin-angiotensin system. Megsin is a novel serine protease inhibitor predominantly expressed in mesanguim. Megsin is upregulated in kidney samples of patients with diabetic nephropathy. Transgenic mice overexpressing megsin spontaneously develop kidney disease characterized by mesangial injury. Megsin is likely to contribute to mesangial injury in the process of diabetic nephropathy. Lack of appropriate animal models has hampered understanding the pathogenesis of diabetic nephropathy and development of effective therapies. Megsin and AGEs are suitable targets for new drugs of diabetic nephropathy and for the development of appropriate animal models of diabetic nephropathy. PMID- 16037284 TI - Effects of advanced glycation end products on ezrin-dependent functions in LLC PK1 proximal tubule cells. AB - We have recently shown that advanced glycation products (AGEs) bind to the ERM (ezrin, radixin, moesin) family of proteins. ERM proteins act as cross-linkers between cell membrane proteins and the actin cytoskeleton. They are also involved in signal transduction pathways. They therefore have a critical role in normal cell processes, including modulation of cell shape, adhesion, and motility. We postulate that AGEs may contribute to diabetic complications by disrupting ERM function. In support of this hypothesis, AGEs inhibit ezrin-dependent tubulogenesis of proximal tubule cells. Phosphorylation is an important activating mechanism for ERM proteins, and AGEs inhibit ezrin phosphorylation mediated by the epidermal growth factor receptor. PMID- 16037285 TI - Localization of the ezrin binding epitope for glycated proteins. AB - ERM proteins (ezrin, radixin, and moesin) have recently been identified as a new class of AGE-binding proteins. ERM proteins link the plasma membrane with the actin cytoskeleton and regulate cell shape, motility, adhesion, and signal transduction. ERM proteins have three structural domains: the N-terminal domain, a coiled midregion, and the C-terminal domain. The N-terminal domain binds to a number of plasma membrane ligands and is involved in signal transduction, while the C-domain binds to actin filaments. Binding studies with isolated structural domains showed that glycated proteins bind to an epitope within the N-terminal domain of ezrin (aa 1-324). It is postulated that some of the cellular effects of AGEs leading to diabetic complications may be mediated by binding to this region of ezrin, thereby interrupting the cross-linking between the plasma membrane and actin cytoskeleton and downstream signaling pathways. Indeed, changes in actin arrangement, cell shape, and adhesion have been described in diabetes, and AGE BSA inhibits ezrin-dependent tubulogenesis of LLC-PK1 proximal tubular cells. For future development of antagonists, further identification of the ezrin-binding epitope for glycated proteins is required. PMID- 16037286 TI - Processing advanced glycation end product-modified albumin by the renal proximal tubule and the early pathogenesis of diabetic nephropathy. AB - Diabetes is characterized by increased quantities of circulating proteins modified by advanced glycation end products (AGEs). Proteins filtered at the glomerulus and presented to the renal proximal tubule are likely to be highly modified by AGEs. The proximal tubule binds, takes up, and catabolizes AGE modified albumin by pathways different from those of unmodified albumin. These differences were examined in polarized, electrically resistant proximal tubular cells grown in monolayer culture. In patients with type 1 diabetes, urinary excretion of a lysosomal enzyme predicted the development of nephropathy. PMID- 16037287 TI - Significance of proximal tubular metabolism of advanced glycation end products in kidney diseases. AB - Advanced glycation end products (AGEs) are formed by the nonenzymatic Maillard reaction between sugars and proteins. Low-molecular weight AGEs are filtered by renal glomeruli and then reabsorbed and metabolized by proximal tubule cells (PTCs). High-molecular weight AGEs are also delivered to PTCs in proteinuric states. In patients with diabetes, AGE generation is increased, and the actions of AGEs on PTCs are likely involved in the pathogenesis of diabetic nephropathy. In patients with chronic renal failure (CRF), reduced renal metabolism of AGEs likely accounts for the accumulation of AGEs in serum, leading to uremic complications including dialysis-related amyloidosis. AGE precursors such as reactive carbonyl compounds also accumulate in the sera of patients with CRF. It is likely that PTCs take up AGEs and AGE precursors via specific endocytotic receptors or transporters. Megalin is a multiligand endocytotic receptor that is abundantly expressed on PTCs. There is evidence that megalin is involved in the cellular uptake and degradation of AGEs. We previously reported a cell therapy model involving implantation of megalin-expressing cells into experimental mice with renal failure for elimination of uremic toxin proteins. Further studies are needed to clarify the molecular mechanisms of the metabolism of AGEs and their precursors to develop a strategy for the treatment of diabetic nephropathy and uremic complications of CRF. PMID- 16037288 TI - Low-molecular weight advanced glycation end products: markers of tissue AGE accumulation and more? AB - Incomplete digestion of advanced glycation end product (AGE)-modified protein results in the formation of low-molecular weight degradation products incorporating AGE modifications (LMW-AGEs). In addition to being biomarkers of AGE modification, LMW-AGEs may have a high toxic potential, being free to interact with AGE receptors at distant sites via the circulation. Several free AGEs have been identified, including pentosidine, N(epsilon) (carboxymethyl)lysine (CML), and free-imidazole AGEs. In addition, fluorescence (370 nm [excitation]/440 nm [emission]) in the LMW phase of serum correlates with tissue fluorescence, an established marker for AGE modification. In experimental diabetes, LMW fluorescence increases with duration of disease and is normalized with the AGE inhibitor aminoguanidine. LMW fluorescence is also higher in patients with diabetes, in whom it is associated with glomerular filtration rate and hemoglobin. Patients with hyperfiltration have lower LMW fluorescence than those with normal renal function, which may protect them from AGE accumulation in the short term. These findings provide clinical support for the association between AGEs and progressive renal injury in diabetes. PMID- 16037289 TI - Plasma low-molecular weight fluorescence in type 1 diabetes mellitus. AB - Characteristic tissue fluorescence is associated with advanced glycation end product (AGE) accumulation in experimental diabetes models, but its utility in patients with type 1 diabetes remains to be established. We studied 148 patients with type 1 diabetes and 77 healthy age-matched control subjects. Low-molecular weight (LMW) fluorophore levels were estimated in plasma samples obtained after an overnight fast. Intra- and interassay coefficients of variation were 4.7% and 6.4%, respectively. LMW fluorophore levels were significantly higher in patients with diabetes than in control subjects (6.3 +/- 0.6 AU/mL vs. 4.1 +/- 0.3; P = 0.007). However, all of this difference came from patients with microvascular complications (n = 67, 7.5 +/- 1.3). There was no significant difference in LMW fluorescence between complication-free patients (4.4 +/- 0.2) and control subjects (P > 0.05). On multivariate analysis, LMW fluorophores correlated with measures of renal function (P < 0.05) but not with diabetes per se. In addition, there was no correlation between LMW fluorophores and the markers of oxidative stress or systemic inflammation. Longitudinal and interventional studies are required to determine whether the association between LMW fluorophores and nephropathy is cause or effect. PMID- 16037290 TI - Plasma levels of AGE peptides in type 1 diabetic patients are associated with serum creatinine and not with albumin excretion rate: possible role of AGE peptide-associated endothelial dysfunction. AB - Patients with renal impairment have an increased risk for cardiovascular disease, which may be the result of advanced glycation end products (AGEs). The aim of this study was to investigate the levels of AGE peptides in relation to kidney function and to study the relationship of AGE peptides with endothelial function and inflammation in type 1 diabetic patients. We measured plasma levels of AGE peptides with a simple fluorescent analytical procedure in patients with end stage renal disease with or without diabetes and in 60 type 1 diabetic patients categorized as having normo-, micro-, or macroalbuminuria. Using enzyme-linked immunosorbent assays, we determined vascular cell adhesion molecule 1 (sVCAM-1), sE-selectin, plasminogen activator inhibitor 1 (PAI-1), tissue type-specific plasminogen activator (tPA), von Willebrand factor (vWF), and soluble thrombomodulin (sTM) to be markers of endothelial function and determined C reactive protein (CRP) to be a marker of inflammation. AGE peptides were increased approximately fivefold in patients with end-stage renal disease, without difference between patients with or without diabetes. In type 1 diabetic patients, the increase of AGE peptides across the groups normo-, micro-, and macroalbuminuria (with medians [range] of 12.6% [7.8-27.2%], 12.1% [7.8-162%], and 46.5% [9.0-248.9%]) was associated with serum creatinine level and not with albumin excretion rate (AER). The relationship with serum creatinine decreased but remained significant after adjusting for age, sex, diabetes duration, hemoglobin A1c (HbA1c), AER, systolic and diastolic blood pressure (BP), and CRP in multiple linear-regression analysis. AGE peptide levels were significantly associated with sVCAM-1 and sTM, independently of serum creatinine. However, these relationships were no longer significant after adjusting for age, sex, diabetes duration, HbA1c, AER, systolic and diastolic BP, and CRP. This study shows that plasma levels of AGE peptides rise with renal impairment, as determined by serum creatinine. AGE peptides are associated with some markers of endothelial activation, which may suggest an involvement of AGE peptides in the acceleration of cardiovascular complications in type 1 diabetic patients with renal impairment. PMID- 16037291 TI - Pathological roles of advanced glycation end product receptors SR-A and CD36. AB - The pathological significance of advanced glycation end product (AGE)-modified proteins deposited in several lesions is generally accounted for by their cellular interaction via the AGE receptors and subsequent acceleration of the inflammatory process. In this study, we focused on two AGE receptors specifically, the role of SR-A in pathogenesis of diabetic nephropathy and the role of CD36 in AGE-induced downregulation of leptin by adipocytes. In terms of SR-A, diabetic wild-type mice exhibited increased urinary albumin excretion, glomerular hypertrophy, and mesangial matrix expansion, whereas SR-A-knockout mice showed reduced glomerular size and mesangial matrix area. In these diabetic SR-A-knockout mice, the number of macrophages that infiltrated into glomeruli was remarkably reduced (P < 0.05), suggesting that SR-A-dependent glomerular migration of macrophages plays an important role in the pathogenesis of diabetic nephropathy. In terms of CD36, incubation of glycolaldehyde-modified bovine serum albumin (GA-BSA) with 3T3-L1 adipocytes reduced leptin secretion by these cells. The binding of GA-BSA to these cells and subsequent endocytic degradation were effectively inhibited by a neutralizing anti-CD36 antibody. AGE-induced downregulation of leptin was protected by N-acetyl-cysteine, an antioxidant. These results indicate that the interaction of AGE ligands with 3T3-L1 adipocytes via CD36 induces oxidative stress and leads to inhibition of leptin expression by these cells, suggesting a potential link of this phenomenon to exacerbation of the insulin sensitivity in metabolic syndrome. PMID- 16037292 TI - Advanced glycation end product receptor-mediated cellular dysfunction. AB - Advanced glycation end products (AGEs), S100/calgranulins, and HMGB1 proteins supposedly play a pivotal role in diabetes mellitus and other chronic inflammatory diseases by promoting cellular dysfunction via binding to cellular surface receptors. Particularly, engagement of the receptor for AGEs (RAGE) has gained major attention because it converts short-lasting cellular activation in sustained cellular dysfunction. Consistently, blockade of ligand-RAGE interaction with soluble RAGE (sRAGE) suppresses chronic cellular activation and dysfunction in animal models of chronic diseases. RAGE-/- mice, however, demonstrate that the protection conferred by RAGE deficiency is lower than that mediated by sRAGE. Furthermore, RAGE-/- mice can be protected by sRAGE in certain settings of the adaptive immune response. This finding implies that abounding RAGE ligands overworking the RAGE pathway might also activate other receptors. PMID- 16037293 TI - Advanced glycation end products: a possible link to angiotensin in an animal model. AB - Advanced glycation end products (AGEs) have been associated with progressive vascular and renal damage in a variety of pathological conditions such as renal failure and diabetes mellitus. The formation of AGEs is generally attributed to increased oxidative and carbonyl stress or hyperglycemia. Activation of the cellular receptor of AGE (RAGE) leads to subsequent cellular activation and proinflammatory responses. Angiotensin (Ang) produces cellular oxidative stress and similarly promotes end organ damage via its type 1 receptor. We investigated the interrelation between these two systems in a new transgenic rat (TGR) model with Ang II-dependent hypertension and renal damage and in nontransgenic controls. TGR showed increased systolic blood pressure (approximately 210 mmHg), proteinuria, and increased renal collagen I mRNA expression compared with normotensive nontransgenic controls. Immunohistochemical staining of kidney sections showed colocalization for Nepsilon-carboxy(methyl)lysine, RAGE, and NF kappaB in TGR glomeruli. These features were absent in nontransgenic controls. Our observations suggest a possible link between Ang II-dependent end-organ damage and the AGE/RAGE axis in vivo. TGRs provide an excellent model to study the interrelation between the renin-angiotensin system and the AGE/RAGE axis in promoting cardiovascular end-organ damage, which would otherwise not be possible in humans. PMID- 16037294 TI - Genotoxicity of advanced glycation end products: involvement of oxidative stress and of angiotensin II type 1 receptors. AB - In patients with chronic renal failure, cancer incidence is increased. This may be related to an elevated level of genomic damage, which has been demonstrated by micronuclei formation as well as by comet assay analysis. Advanced glycation end products (AGEs) are markedly elevated in renal failure. In the comet assay, the model AGEs methylglyoxal- and carboxy(methyl)lysine-modified bovine serum albumin (BSA) induced significant DNA damage in colon, kidney, and liver cells. The addition of antioxidants prevented AGE-induced DNA damage, suggesting enhanced formation of reactive oxygen species (ROS). The coincubation with dimethylfumarate (DMF), an inhibitor of NF-kappaB translocation, reduced the genotoxic effect, thereby underscoring the key role of NF-kappaB in this process. One of the genes induced by NF-kappaB is angiotensinogen. The ensuing proteolytic activity yields angiotensin II, which evokes oxidative stress as well as proinflammatory responses. A modulator of the renin-angiotensin system (RAS), the angiotensin II (Ang II) receptor 1 antagonist, candesartan, yielded a reduction of the AGE-induced DNA damage, connecting the two signal pathways, RAS and AGE signaling. We were able to identify important participants in AGE-induced DNA damage: ROS, NF-kappaB, and Ang II, as well as modulators to prevent this DNA damage: antioxidants, DMF, and AT1 antagonists. PMID- 16037295 TI - Glycolaldehyde-modified bovine serum albumin downregulates leptin expression in mouse adipocytes via a CD36-mediated pathway. AB - Previous observations by us have clarified that proteins modified by advanced glycation end products (AGEs) are recognized as effective ligands by CD36 overexpressed CHO cells and undergo receptor-mediated endocytosis. CD36, a member of the class B scavenger receptor family, also acts as a fatty acid transporter in adipocytes. Oxidized low-density lipoprotein (Ox-LDL), a ligand for CD36, is known to upregulate CD36 by activating peroxisome proliferator-activated receptor gamma (PPAR-gamma) in macrophages, whereas PPAR-gamma ligands such as troglitazone and 15-deoxy-delta12,14-prostaglandin J2 decrease leptin secretion from adipocytes. The purpose of this study was to examine effects of AGE ligands on leptin expression in adipocytes. Glycolaldehyde-modified bovine serum albumin (GA-BSA) decreased leptin expression at both the protein and mRNA levels in 3T3 L1 adipocytes and mouse epididymal adipocytes. The binding to and subsequent endocytic degradation of GA-BSA by 3T3-L1 adipocytes were effectively inhibited by a neutralizing anti-CD36 antibody. These results indicate that the ligand interaction of GA-BSA with CD36 leads to downregulation of leptin expression in 3T3-L1 adipocytes, suggesting that AGE-induced leptin downregulation is linked to reduction of the insulin sensitivity in metabolic syndrome. PMID- 16037296 TI - Aldose reductase and AGE-RAGE pathways: key players in myocardial ischemic injury. AB - Cardiovascular disease represents the major cause of morbidity and mortality in patients with diabetes mellitus. The impact of cardiac disease includes increased sensitivity of diabetic myocardium to ischemic episodes and diabetic cardiomyopathy, manifested as a subnormal functional response of the diabetic heart independent of coronary artery disease. In this context, we were to our knowledge the first to demonstrate that diabetes increases glucose flux via the first and key enzyme, aldose reductase, of the polyol pathway, resulting in impaired glycolysis under normoxic and ischemic conditions in diabetic myocardium. Our laboratory has been investigating the role of the polyol pathway in mediating myocardial ischemic injury in diabetics. Furthermore, the influence of the aldose reductase pathway in facilitating generation of key potent glycating compounds has led us to investigate the impact of advanced glycation end products (AGEs) in myocardial ischemic injury in diabetics. The potent impact of increased flux via the aldose reductase pathway and the increased AGE interactions with its receptor (RAGE) resulting in cardiac dysfunction will be discussed in this chapter. PMID- 16037297 TI - Advanced glycation end products and bone loss during aging. AB - It is well known that bone mass density decreases with age. Age-related bone mass loss is ascribed to several factors. Nonenzymatic glycation has been proposed as a new potential factor in the loss of bone during aging. In this study we evaluated the concentration of pentosidine, an advanced glycation end product, in cortical and trabecular bone and in the plasma of subjects undergoing orthopedic surgery. The relationship between these parameters and a clinical index of osteoporosis was also studied. Samples of bone and plasma of 104 nondiabetic subjects (74 women and 30 men), 72 +/- 1 years old, were studied. Pentosidine was determined by HPLC after decalcification and hydrolysis. The radiologic Singh index was evaluated blindly by orthopedic surgeons to provide the degree of osteoporosis. Pentosidine concentration of cortical bone shows a significant exponential increase with age (r = 0.610, P < 0.001). This increase, however, is not seen in the trabecular bone, which is characterized by a large spread in the data. Interestingly the concentration of cortical pentosidine is also related to the Singh score (r(s) = -0.274, P < 0.01). Plasma pentosidine has a significant exponential correlation with age (r = +0.339, P < 0.001) and a linear correlation with the cortical bone pentosidine (r = +0.248, P < 0.05). This study demonstrates that pentosidine increases exponentially in cortical bone during aging, and is thus a good biomarker for the degree of bone mass density loss. The trabecular bone concentration of pentosidine is more variable, probably because of the turnover rate and the local environment; plasma pentosidine might provide information on the bone turnover rate. PMID- 16037298 TI - Increased protein glycation in cirrhosis and therapeutic strategies to prevent it. AB - Glycation of liver proteins by reactive aldehydes formed from the metabolism of ethanol and lipid peroxidation has been implicated in the development of both alcoholic and nonalcoholic liver cirrhosis. Modified proteins are targeted to the proteasome for proteolysis. Release of glycation-free adducts into the circulation may provide a diagnostic "signature" of hepatic protein damage. We quantitatively screened protein glycation, oxidation, and nitrosation adduct residues and free adducts in portal, hepatic, and peripheral venous blood plasma of cirrhotic patients; we also screened the hepatic and peripheral venous blood plasma of control subjects by liquid chromatography-mass spectrometry. There was a remarkable 14-16-fold increase of glyoxal-derived, hydroimidazolone-free adduct in portal and hepatic venous plasma of cirrhotic patients with respect to normal controls. There was only a twofold increase of glycation adduct residues in plasma proteins in cirrhotic patients, which was attributed mainly to decreased albumin turnover. Therapeutic strategies to decrease dicarbonyl compounds may be beneficial, such as dicarbonyl scavengers, glutathione repleting agents, and high dose thiamine therapy. PMID- 16037299 TI - Advanced glycation end products in human cancer tissues: detection of Nepsilon (carboxymethyl)lysine and argpyrimidine. AB - Tumors are generally characterized by an increased glucose uptake and a high rate of glycolysis. Since one consequence of an elevated glycolysis is the nonenzymatic glycation of proteins, we studied the presence of advanced glycation end products (AGEs) in human cancer tissues. We detected the presence of the AGEs N(epsilon)-(carboxymethyl)lysine (CML) and argpyrimidine in several human tumors using specific antibodies. Because AGEs have been associated with the etiology of a variety of different diseases, these results suggest that CML and argpyrimidine could be implicated in the biology of human cancer. PMID- 16037300 TI - Advanced glycation end product free adducts are cleared by dialysis. AB - Plasma advanced glycation end product (AGE) free adducts are increased up to 50 fold among patients on dialysis. We examined the ability of hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) to clear these compounds. The AGE free adducts Nepsilon-carboxymethyl-lysine (CML) and Nepsilon-(1 carboxyethyl)lysine (CEL) and the hydroimidazolones derived from glyoxal (G-H1), methylglyoxal (MG-H1), and 3-deoxyglucosone (3DG-H) were determined by LC-MS/MS and pentosidine by HPLC with fluorimetric detection in ultrafiltrates of plasma, urine, or PD effluent as appropriate from patients on HD (n = 8) or PD (n = 8), and from healthy controls (n = 8). Among patients on HD, all free AGEs predialysis were significantly higher than in controls and were decreased with dialysis. The removal of MG-H1 and 3DG-H was comparable to that of urea, whereas that of CML and pentosidine was some 20% higher; in contrast, the removal of CEL and G-H1 was 25% lower. Among patients on CAPD, free AGEs in PD effluent increased with increasing dwell time. The combined renal and peritoneal 24-h excretion rates of CML (4.7 micromol), CEL (6.5 micromol), 3DG-H (16.6 micromol), and pentosidine (0.08 micromol) were twofold higher than the amount excreted in healthy controls, whereas MG-H1 was ninefold higher (59 micromol); the combined clearances of all free AGEs except pentosidine were lower than in healthy controls. Impaired renal clearance contributes to increased plasma free AGEs in uremia, but the increased excretion rate among patients on PD demonstrates that there was also an increased synthesis of free AGEs. Both HD and PD are able to remove free AGEs. PMID- 16037301 TI - From molecular footprints of disease to new therapeutic interventions in diabetic nephropathy. AB - Proteins are particularly attractive targets for product analysis, which is used to understand pathology. Protein modifications, such as advanced glycation end products (AGEs), serve as footprints of biochemical processes and also help in the search for novel agents that efficiently inhibit protein damage. Interestingly, several medical agents that are used clinically interfere with oxidative protein damage through different mechanisms characteristic of their chemical structures. We recently found that angiotensin II receptor blockers (ARBs) and angiotensin converting enzyme inhibitors (ACEIs) lower the in vitro formation of the AGEs pentosidine and carboxymethyllysine. Their inhibition for AGE formation is more striking than aminoguanidine. Unlike aminoguanidine, ARBs and ACEIs do not trap reactive carbonyl precursors of AGEs. Rather, they inhibit AGE formation, possibly as a result of their potent ability to scavenge hydroxyl radicals and to chelate the transition metals necessary for the Fenton reaction. We tested their AGE-lowering ability in vivo in a unique type-2 diabetic model with nephropathic SHR/NDmcr-cp rats, which exhibits the metabolic syndrome (obesity, hyperglycemia, hyperlipidemia, hyperinsulinemia) in addition to hypertension. Obesity and associated metabolic derangements, in addition to hypertension, markedly accelerate renal injury. Expectedly, correction of hyperglycemia and hyperinsulinemia partially but significantly improves renal injury. A low-calorie diet greatly improves renal injury despite persistent hypertension. Among antihypertensive agents, ARBs, unlike nifedipine and atenolol, are renoprotective despite persistent metabolic syndrome, but their action is independent of blood pressure lowering and is observed in a dose dependent manner despite the complete blockade of angiotensin II receptor. Interestingly, the improvement of renal injury by ARBs as well as a low-calorie diet is associated with a significant reduction in local oxidative stress and AGE formation in the kidney. During the characterization of the AGE-lowering profile of our chemical compound libraries ( approximately 2000), we identified several inhibitors of oxidative stress and advanced glycation. They are indeed renoprotective, independently of correction of hypertension and metabolic syndrome, in experimental diabetic nephropathy and other nephritis models. Altogether, our data are in good agreement with the recent therapeutic concept for diabetic nephropathy that multiple risk factor interventions are critical in the treatment of diabetic renal injury, and further implicate a therapeutic potential of inhibition of oxidative stress and advanced glycation. PMID- 16037302 TI - Can advanced glycation end product inhibitors modulate more than one pathway to enhance renoprotection in diabetes? AB - Although advanced glycation end products (AGEs) have been postulated to contribute to diabetic nephropathy in their own right, advanced glycation is clearly only one pathway by which renal injury may be induced in diabetes. The interaction between metabolic and hemodynamic factors amplifies the deleterious effects of the diabetic milieu, thereby reducing the threshold for microvascular injury via common mechanisms. This includes interactions between AGE-mediated pathways and the renin angiotensin system, oxidative stress, protein kinase C, and growth factors, which play a significant role in the development and progression of diabetic renal disease. As it is likely that the future of preventive therapy will not involve a single "cure-all" agent, it seems that a highly relevant question in diabetic nephropathy should be, which pathogenic pathways are already addressed by currently available therapies? Combination therapies that target multiple pathways may ultimately be more successful than those that modify a single pathway. Therefore, research into synergistic interactions among the various pathogenic pathways leading to diabetic complications is critical in order to develop interventions that confer optimal end-organ protection. PMID- 16037303 TI - Advanced glycation end products in diabetes-associated atherosclerosis and renal disease: interventional studies. AB - There is increasing evidence that advanced glycation end products (AGEs) and their interactions with various receptors (in particular, the receptor RAGE) play a pivotal role in the development and progression of diabetic macro- and microvascular complications. Several approaches have been used to inhibit tissue accumulation of AGEs in diabetes, including inhibitors of AGE formation such as aminoguanidine, ALT 946, and pyridoxamine-or putative cross-link breakers such as ALT 711. Alternative interventions have also included the administration of a soluble receptor for RAGE, sRAGE, thus capturing circulating AGEs and preventing them from binding to the cell-bound full-length receptor RAGE, thereby inhibiting the proinflammatory and profibrotic response following AGE-RAGE binding. In this review we summarize the evidence for such antiglycation therapies in retarding or delaying the development and progression of diabetes-associated atherosclerosis and renal disease while focusing on interventional strategies inhibiting AGE accumulation. In summary, all approaches have been shown to confer some degree of antiatherosclerotic and renoprotective effects, albeit to different degrees and by different mechanisms. PMID- 16037304 TI - Renoprotective and lipid-lowering effects of LR compounds, novel advanced glycation end product inhibitors, in streptozotocin-induced diabetic rats. AB - The accelerated formation of advanced glycation/lipoxidation end products (AGEs/ALEs) has been implicated in the pathogenesis of various diabetic complications. Several natural and synthetic compounds have been proposed and advanced as inhibitors of AGE/ALE formation. We examined the effects of two new AGE/ALE inhibitors, LR-9 and LR-74, on the prevention of early renal disease and dyslipidemia in streptozotocin (STZ)-induced diabetic rats. Diabetic rats were treated with either LR-9 or LR-74 for 32 weeks. Progression of renal disease was evaluated by measurements of urinary albumin and plasma creatinine concentrations. AGE-induced chemical modification of the tail tendon collagen and levels of Nepsilon-(carboxymethyl)- and (carboxyethyl)- lysines (CML and CEL) in skin collagen were measured. AGE/ALE levels in kidneys were determined by immunohistochemistry. Plasma lipids and their lipid hydroperoxide concentrations were also determined. Treatment of either LR-9 or LR-74 significantly inhibited the increase in albuminuria, plasma creatinine, hyperlipidemia, and plasma lipid peroxidation in diabetic rats without any effects on hyperglycemia. Both compounds also reduced CML-AGE accumulation in kidney glomeruli and tubules, AGE linked fluorescence and cross-linking of tail collagen, and levels of CML and CEL in skin collagen. These results suggest that both LR compounds can inhibit the progression of renal disease and also prevent dyslipidemia in experimental diabetes. These compounds may have an additional beneficial effect as an antioxidant against lipid peroxidation, and thus may provide alternative therapeutic options for the treatment of various diabetic macrovascular complications. PMID- 16037305 TI - High-dose thiamine therapy counters dyslipidemia and advanced glycation of plasma protein in streptozotocin-induced diabetic rats. AB - The streptozotocin-induced (STZ) diabetic rat experimental model of diabetes on insulin maintenance therapy exhibits dyslipidemia, mild thiamine deficiency, and increased plasma protein advanced glycation end products (AGEs). The reversal of thiamine deficiency by high-dose thiamine and S-benzoylthiamine monophosphate (benfotiamine) prevented the development of incipient nephropathy. Recently, we reported that high-dose thiamine (but not benfotiamine) countered diabetic dyslipidemia. To understand further the differences between the effects of thiamine and benfotiamine therapy, we quantified the levels of the AGEs in plasma protein. We found hydroimidazolone AGE residues derived from glyoxal and methylglyoxal, G-H1 and MG-H1, were increased 115% and 68% in STZ diabetic rats, with respect to normal controls, and were normalized by both thiamine and benfotiamine; whereas N-carboxymethyl-lysine (CML) and N-carboxyethyl-lysine (CEL) residues were increased 74% and 118% in STZ diabetic rats and were normalized by thiamine only. The lack of effect of benfotiamine on plasma CML and CEL residue concentrations suggests there may be important precursors of plasma protein CML and CEL residues other than glyoxal and methylglyoxal. These are probably lipid-derived aldehydes. PMID- 16037306 TI - Inhibitors of advanced glycation end product formation and neurovascular dysfunction in experimental diabetes. AB - Advanced glycation and lipoxidation end products (AGEs/ALEs) have been implicated in the pathogenesis of the major microvascular complications of diabetes mellitus: nephropathy, neuropathy, and retinopathy. This article reviews the evidence regarding the peripheral nerve and its vascular supply. Most investigations done to assess the role of AGEs/ALEs in animal models of diabetic neuropathy have used aminoguanidine as a prototypic inhibitor. Preventive or intervention experiments have shown treatment benefits for motor and sensory nerve conduction velocity, autonomic nitrergic neurotransmission, nerve morphometry, and nerve blood flow. The latter depends on improvements in nitric oxide-mediated endothelium-dependent vasodilation and is responsible for conduction velocity improvements. A mechanistic interpretation of aminoguanidine's action in terms of AGE/ALE inhibition is made problematic by the relative lack of specificity. However, other unrelated compounds, such as pyridoxamine and pyridoxamine analogues, have recently been shown to have beneficial effects similar to aminoguanidine, as well as to improve pain-related measures of thermal hyperalgesia and tactile allodynia. These data also stress the importance of redox metal ion-catalyzed AGE/ALE formation. A further approach is to decrease substrate availability by reducing the elevated levels of hexose and triose phosphates found in diabetes. Benfotiamine is a transketolase activator that directs these substrates to the pentose phosphate pathway, thus reducing tissue AGEs. A similar spectrum of improvements in nerve and vascular function were noted when using benfotiamine in diabetic rats. Taken together, the data provide strong support for an important role for AGEs/ALEs in the etiology of diabetic neuropathy. PMID- 16037307 TI - Post-Amadori AGE inhibition as a therapeutic target for diabetic complications: a rational approach to second-generation Amadorin design. AB - Aminoguanidine and pyridoxamine (Pyridorintrade mark), two major inhibitors of advanced glycation end product (AGE) formation, have entered clinical trials for diabetic nephropathy. They share no structural similarity and are believed to inhibit AGE formation by entirely different mechanisms. Pyridoxamine is a post Amadori AGE inhibitor-that is, an "Amadorin"-whereas aminoguanidine primarily scavenges reactive dicarbonyl precursors to AGEs. However, pyridoxamine also has a limited potential to react with dicarbonyls. We thus embarked on an effort to develop second-generation Amadorins with low nucleophilicity. Our hypothesis was that we could improve specificity for inhibiting the post-Amadori pathway by minimizing the potential for scavenging small dicarbonyl intermediates. This mechanism-based strategy has led to a rational drug design program that has successfully produced candidate Amadorins, among them the novel compound BST 4997. This Amadorin has greater post-Amadori potency than pyridoxamine but possess no dicarbonyl scavenging activity. Prototypical inhibitors like BST-4997 provide a unique tool to help identify relevant AGE pathways that contribute to diabetic complications. Targeting AGE inhibition differs significantly from traditional approaches to drug discovery and thus represents a new paradigm for the drug industry that should be recognized. PMID- 16037308 TI - Pyridoxamine: the many virtues of a maillard reaction inhibitor. AB - Pyridoxamine (PM) is one of three natural forms of vitamin B6. It is a critical transient intermediate in catalysis of transamination reactions by vitamin B6 dependent enzymes. The discovery eight years ago that PM can inhibit the Maillard reaction stimulated new interest in this B6 vitamer as a prospective pharmacological agent for treatment of complications of diabetes. PM application in diabetic nephropathy has now progressed to a phase III clinical trial. Investigation of the PM mechanism of action demonstrated that PM inhibits post Amadori steps of the Maillard reaction by sequestering catalytic metal ions and blocking oxidative degradation of Amadori intermediate. PM also has the capacity to scavenge toxic carbonyl products of sugar and lipid degradation, and to inhibit reactive oxygen species. These multiple activities position PM as a promising drug candidate for treatment of multifactorial chronic conditions in which oxidative reactions and/or carbonyl compounds confer pathogenicity. PMID- 16037309 TI - Tissue-specific variation in glycation of proteins in diabetes: evidence for a functional role of amadoriase enzymes. AB - The Amadori product fructoselysine (FL), an intermediate in the formation of many advanced glycation end products, may be deglycated by various pathways. These include spontaneous chemical degradation or enzymatic deglycation by amadoriases. This study was designed to compare changes in FL in various tissues in response to changes in glycemia, thereby testing tissue-specific deglycation. FL content in skin collagen, red cell hemoglobin, and total muscle, liver, and brain protein was analyzed by isotope dilution gas chromatography-mass spectrometry. Mean blood glucose increased over fourfold in diabetic versus control rats, whereas changes in glycation of proteins varied from fivefold in collagen to no change in the liver and brain. These results suggest significant differences among tissues in the activity of deglycating enzymes and/or protein turnover. PMID- 16037310 TI - Some clues as to the regulation, expression, function, and distribution of fructosamine-3-kinase and fructosamine-3-kinase-related protein. AB - Fructosamine-3-kinase (FN3K) and the more recently discovered fructosamine-3 kinase-related protein (FN3KRP) appear to protect proteins from nonenzymatic glycation. To gain a better understanding of these enzymes we performed a series of investigations including (1) in silico comparisons of their promoters; (2) real-time PCR analysis of their expression in human tissues; (3) effects of hyperglycemia, interleukin-1beta (IL-1beta), and nuclear factor kappa-B (NFkappaB) activation on their mRNA levels; (4) effects of small interfering RNA (siRNA) suppression of FN3K expression (knockdown) in cultured cells and (5) search of FN3K and FN3KRP homologs in available genomic and EST (expressed sequence tag) databases. Our results indicate that (1) both FN3K and FN3KRP promoters are TATA-less and CAAT-less and contain several homologous CpG islands and Sp1 binding sites. (2) Both genes are expressed in all human tissue examined, with FN3K showing significantly higher levels in tissues susceptible to nonenzymatic glycation and diabetic complications. (3) Treatment of fibroblasts with high glucose, IL-1beta, and activation of NFkappaB does not affect the expression of either FN3K or FN3KRP. (4) Knockdown of FN3K in cultured cells inhibits or arrests their growth. (5) FN3K-like genes are widely distributed in nature, with the notable exception of insects and yeasts. These data suggest that FN3K and FN3KRP are constitutive "housekeeping" genes and that they play an important role in cell metabolism, possibly as deglycating enzymes. PMID- 16037311 TI - Substrate specificity of amadoriase I from Aspergillus fumigatus. AB - Generation of Amadori products is the major single modification by the Maillard reaction in vivo and a source of biologically active glycoxidation products leading to protein cross-linking in biological tissues. Amadoriase I from Aspergillus fumigatus cleaves Amadori products into deoxyglucosone, hydrogen peroxide, and the corresponding primary amine. It has been reported that Amadori products formed on free amino acids are a good substrate for amadoriase I, whereas the enzyme is unable to cleave Amadori products formed on whole proteins. This work aims to investigate the affinity of amadoriase I for oligopeptides and small proteins. Recombinant amadoriase I was expressed in E. coli and purified by Ni His-tag affinity chromatography. Di-, tri-, and tetrapeptides were derivatized with glucose, and the corresponding Amadori products were purified by TLC and HPLC. Glycated beta-lactoglobulin was also used as a substrate. In both cases the formation of Amadori products was monitored by electrospray ionization-mass spectrometry (ESI-MS). The Km of amadoriase for glycated-L-lysine was 4.2 mM, which is in accordance with the literature. Km decreases with the length of peptide, being slightly reduced for dipeptides, and is around 10 mM for tri- and tetrapeptides. Glycated proteins are not substrates of the enzyme; but when amadoriase I was added during the glycation reaction, a significant reduction of Amadori product formation was observed on both peptides and proteins. Data confirm the hypothesis that steric hindrance is critical for amadoriase I activity, indicating that oligopeptides up to four amino acids in length are good substrates. Moreover, these data show that, at least in some experimental conditions, amadoriase I is able to reduce the formation of protein-bound Amadori product. PMID- 16037312 TI - Transglycation--a potential new mechanism for deglycation of Schiff's bases. AB - Nonenzymatic glycation is believed to play a major role in the development of diabetic complications. Over the past several years we and others have shown that in cells this nonenzymatic process can be reversed by an ATP-dependent reaction catalyzed by fructosamine-3-kinase (FN3K) and possibly by its isozyme, fructosamine-3-kinase-related protein (FN3KRP). In this study we provide the first evidence that this FN3K-dependent deglycation, acting on the Amadori products, is complemented by another deglycation process operating on the very first product of nonenzymatic glycation, glucosylamines (Schiff's bases). We postulate that the first step in this Schiff's-base deglycation process occurs by transfer of the sugar moiety from macromolecule-bound glucosylamine to one of the low-molecular weight intracellular nucleophiles-in particular, glutathione. We term this reaction transglycation, and in this study we demonstrate that it occurs readily and spontaneously in vitro. We further propose that one of the spontaneously formed glucose-glutathione adduct(s) is subsequently removed from cells by a multidrug-resistance pump (MRP, MDR-protein, ATP-binding-cassette protein), metabolized, and excreted in urine. In support of this latter contention, we show that at least one transglycation product, glucose-cysteine, is found in human urine and that its concentrations are increased in diabetes. PMID- 16037313 TI - Proteins of Thermus thermophilus are resistant to glycation-induced protein precipitation: an evolutionary adaptation to life at extreme temperatures? AB - In thermophilic bacteria, formation of Maillard products may occur at increased rates because this reaction is favored at higher temperatures. Therefore, specific protective mechanisms against glycation-induced protein precipitation are likely to exist in thermophilic bacteria. Indeed, Thermus thermophilus proteins remained soluble when a cell-free extract of T. thermophilus was incubated at 37 degrees C in the presence of glucose, fructose, or methylglyoxal; whereas E. coli proteins precipitated. In E. coli cell-free extracts, sugar induced precipitation was accelerated by the addition of 5 microM Fe2+ and inhibited by metal chelators, suggesting that glycoxidation processes are involved in the formation of the precipitate. A low lysine content, endogenous small scavenger molecules, or enzymatic "antiglycation" mechanisms for the degradation of AGEs or their precursors could be excluded as possible causes for the resistance to protein precipitation in T. thermophilus. Therefore, the resistance to glycation-mediated protein precipitation is an endogenous property of thermophilic proteins that was acquired during evolution in environments with high glycation activity. PMID- 16037314 TI - Maillard reaction products derived from thiol compounds as inhibitors of enzymatic browning of fruits and vegetables: the structure-activity relationship. AB - Some thiol-derived Maillard reaction products (MRPs) may exert antioxidant activity, depending on the reaction conditions as well as on the sugar and the sulphydryl compound. Recently, we reported that MRPs derived from glucose or fructose with cysteine (CSH) or glutathione (GSH) mixtures greatly inhibited polyphenoloxidases (PPOs), oxidoreductases responsible for discoloration of fresh or minimally processed fruits and vegetables. Glucose and GSH were shown to be the most active in producing inhibitory MRPs. Therefore, we examined the way in which the nature of the reactants affected their synthesis, in order to establish a structure-activity relationship for the inhibitory products. Various aqueous (0.083 M, 0.125 M, or 0.25 M) mixtures of a sugar (hexose, pentose, or diholoside) with either a CSH-related compound (CSH, GSH, N-acetyl-cysteine, cysteamine, cysteic acid, methyl-cysteine, cysteine methyl ester), an amino acid (gamma-glutamic acid, glycine, methionine), or other sulfur compound (thiourea, 1,4-dithiothreitol, 2-mercaptoethanol) were heated at 103 degrees C for 14 h. Soluble MRPs were compared for their ability to inhibit apple PPO activity. In the presence of CSH, the rated sugars (same molar concentration) ranked as to inhibitory effect were pentoses > sucrose > hexoses > or = maltose. In the presence of glucose, the simultaneous presence of an amino group, a carboxyl group, and a free thiol group on the same molecule seemed essential for the production of highly inhibitory compounds. PMID- 16037377 TI - Expression of insulin-like factor 3 protein in the rat testis during fetal and postnatal development and in relation to cryptorchidism induced by in utero exposure to di (n-Butyl) phthalate. AB - Cryptorchidism is a common reproductive abnormality, possibly resulting from abnormal hormone production/action by the fetal testis. Insulin-like factor 3 (Insl3) is thought to be involved in gubernaculum development and transabdominal testicular descent, but its importance is unclear, due partly to lack of suitable Insl3 antibodies. We generated (by genetic immunization) and validated a novel antirat Insl3 antibody, which we used to characterize immunoexpression of Insl3 in rat Leydig cells (LCs) from fetal life until adulthood and its relationship to cryptorchidism. Immunoexpression was strong on embryonic day (E) 17.5 and E19.5 and from 35 d of age onward but weak from E21.5 until puberty. Because in utero exposure to di (n-butyl) phthalate (DBP) induces cryptorchidism and suppresses Insl3 gene expression, we investigated Insl3 protein expression in fetal and adult rats exposed to 500 mg/kg.d DBP from E13.5 to E21.5. Expression on E17.5 and E19.5 decreased dramatically after DBP exposure, but there was no consistent correlation between this suppression and abnormal testis position. We also compared expression of Insl3 and P450 side-chain cleavage enzyme in fetal testes from rats exposed in utero to DBP or flutamide (50 mg/kg.d). DBP treatment suppressed expression of both P450 side-chain cleavage enzyme and Insl3 at E19.5, but flutamide exposure had no effect on either protein, demonstrating that Insl3 expression in fetal rat LCs is not androgen regulated. In adult rats, Insl3 expression was suppressed in 80% of cryptorchid and 50% of scrotal testes from rats exposed to DBP, suggesting that prenatal DBP exposure also leads to maldevelopment/malfunction of the adult LC population in some animals. PMID- 16037378 TI - Urocortins of the South African clawed frog, Xenopus laevis: conservation of structure and function in tetrapod evolution. AB - Several corticotropin-releasing factor (CRF) family genes have been identified in vertebrates. Mammals have four paralogous genes that encode CRF or the urocortins 1, 2, and 3. In teleost fishes, a CRF, urotensin I (a fish ortholog of mammalian urocortin 1) and urocortin 3 have been identified, suggesting that at least three of the four mammalian lineages arose in a common ancestor of modern bony fishes and tetrapods. Here we report the isolation of genes orthologous to mammalian urocortin 1 and urocortin 3 from the South African clawed frog, Xenopus laevis. We characterize the pharmacology of the frog peptides and show that X. laevis urocortin 1 binds to and activates the frog CRF1 and CRF2 receptors at picomolar concentrations. Similar to mammals, frog urocortin 3 is selective for the CRF2 receptor. Only frog urocortin 1 binds to the CRF-binding protein, although with significantly lower affinity than frog CRF. Both urocortin genes are expressed in brain, pituitary, heart, and kidney of juvenile frogs; urocortin 1 is also expressed in skin. We also identified novel urocortin sequences in the genomes of pufferfish, zebrafish, chicken, and dog. Phylogenetic analysis supports the view that four paralogous lineages of CRF-like peptides arose before the divergence of the actinopterygian and sarcopterygian fishes. Our findings show that the functional relationships among CRF ligands and binding proteins, and their anorexigenic actions mediated by the CRF2 receptor, arose early in vertebrate evolution. PMID- 16037379 TI - Insulin-like growth factor-I receptor signaling in tamoxifen-resistant breast cancer: a supporting role to the epidermal growth factor receptor. AB - There is considerable evidence that the epidermal growth factor receptor (EGFR) and IGF-I receptor (IGF-IR) cross-talk in breast cancer cells. In the present study, we have examined whether EGFR/IGF-IR cross-talk exists in EGFR-positive tamoxifen-resistant variants of MCF-7 (Tam-R) and T47D (T47D-R) breast cancer cell lines. Although Tam-R cells expressed reduced IGF-IR protein levels compared with their wild-type MCF-7 counterparts, phosphorylated IGF-IR protein levels were equivalent in the two cell lines under basal growth conditions, possibly as a consequence of increased IGF-II expression in Tam-R cells. IGF-II activated both IGF-IR and EGFR in Tam-R cells, whereas only activation of IGF-IR was observed in wild-type cells. In contrast, epidermal growth factor rapidly induced EGFR, but not IGF-IR, phosphorylation in Tam-R cells. IGF-II promoted direct association of c-SRC with IGF-IR, phosphorylated c-SRC, and increased EGFR phosphorylation at tyrosine 845, a c-SRC-dependent phosphorylation site. Pretreatment with either AG1024 (IGF-IR-specific inhibitor) or an IGF-II neutralizing antibody inhibited basal IGF-IR, c-SRC, and EGFR phosphorylation, and AG1024 significantly reduced Tam-R basal cell growth. The c-SRC inhibitor SU6656 also inhibited growth, reduced basal and IGF-II-induced c-SRC and EGFR phosphorylation, and blocked EGFR activation by TGFalpha. Similarly, in T47D-R cells, AG1024 and SU6656 inhibited basal and IGF-II-induced phosphorylation of c SRC and EGFR, and SU6656 reduced TGFalpha-induced EGFR activity. These results suggest the existence of a unidirectional IGF-IR/EGFR cross-talk mechanism whereby IGF-II, acting through the IGF-IR, regulates basal and ligand-activated EGFR signaling and cell proliferation in a c-SRC-dependent manner in Tam-R cells. This cross-talk between IGF-IR and EGFR is not unique to Tam-R cells because this mechanism is also active in a tamoxifen-resistant T47D-R cell line. PMID- 16037380 TI - ObRa and ObRe are differentially expressed in adipose tissue in aged food restricted rats: effects on circulating soluble leptin receptor levels. AB - In rodents, soluble leptin receptor (SLR) may be generated by alternative splicing of ObR mRNA and/or as a cleavage product of ObR membrane-anchored receptors. In this study, we investigated the contribution of both processes on the generation of SLR in 3-, 8-, and 24-month-old Wistar rats fed ad libitum (AL) or under food restriction (FR). To this end, we determined serum SLR levels and analyzed ObRa and ObRe mRNA expression under these physiological conditions. Additionally, we studied the cellular distribution of ObRa and the generation of SLR by N-ethyl-maleimide-induced shedding from ObRa membrane receptors in isolated adipocytes. Serum SLR levels were significantly increased in 8- and 24 month-old rats under FR, whereas similar amounts were found in rats of different ages fed AL. ObRa and ObRe mRNA expression in epididymal adipose tissue increased with aging. In contrast, after FR, ObRe mRNA expression decreased, whereas ObRa mRNA expression further increased compared with 8- and 24-month-old rats fed AL. Additionally, FR promoted a change in the distribution of ObRa between internal and plasma membranes in isolated adipocytes, increasing its presence at the cell surface. Finally, the generation of SLR by N-ethyl-maleimide-induced shedding from ObRa was also increased under FR. These data suggest that shedding of ObRa membrane-anchored receptors, rather than ObRe expression, might preferentially contribute to the generation of the increased levels of SLR in serum under conditions of FR. PMID- 16037381 TI - Role of sulfated tyrosines of thyroglobulin in thyroid hormonosynthesis. AB - Our previous studies showed that sulfated tyrosines (Tyr-S) are involved in thyroid hormone synthesis and that Tyr(5), the main hormonogenic site of thyroglobulin (Tg), is sulfated. In the present paper, we studied the role of Tyr S in the formation and activity of the peroxidase-Tg complex. Results show that noniodinated (35)SO(3)-Tg specifically binds (Kd=1.758 microM) to immobilized lactoperoxidase (LPO) via Tyr-S linkage by using saturation binding and competition experiments. We found that NIFEY-S, a 15-amino acid peptide corresponding to the NH2-end sequence of Tg and containing the hormonogenic acceptor Tyr5-S, was a better competitor than cholecystokinin and Tyr-S. 35SO3 Tg, iodinated without peroxidase, bound to LPO with a Kd (1.668 microM) similar to that of noniodinated Tg, suggesting that 1) its binding occurs via Tyr-S linkage and 2) Tyr-S requires peroxidase to be iodinated, whereas nonsulfated Tyr does not. Iodination of NIFEY-S with [125I]iodide showed that Tyr5-S iodination increased with LPO concentration, whereas iodination of a nonsulfated peptide containing the donor Tyr130 was barely dependent on LPO concentration. Enzymatic hydrolysis of iodinated Tg or NIFEY-S showed that the amounts of sulfated iodotyrosines also depended on LPO amount. Sulfated iodotyrosines were detectable in the enzyme-substrate complex, suggesting they have a short life before the coupling reaction occurs. Our data suggest that after Tyr-S binding to peroxidase where it is iodinated, the sulfate group is removed, releasing an iodophenoxy anion available for coupling with an iodotyrosine donor. PMID- 16037382 TI - The adapter protein GRB10 is an endogenous negative regulator of insulin-like growth factor signaling. AB - The growth factor IGF-I is critical for normal human somatic growth and development. Growth factor receptor-bound protein (Grb)10 is a protein that interacts with the IGF-I receptor and may thus regulate IGF-I-stimulated growth. However, the role of endogenous Grb10 in regulating IGF-I action is not known. The objective of this study was to determine the function of endogenous Grb10 in IGF signaling responses. Using small interfering RNA, we demonstrate that knockdown of Grb10 enhances IGF-I-mediated phosphorylation of insulin receptor substrate proteins, Akt/protein kinase B, and ERK1/2 and leads to a corresponding increase in DNA synthesis. Although IGF-I receptor autophosphorylation normally correlates with receptor signaling, we demonstrate a decrease in IGF-I-stimulated receptor phosphorylation in Grb10 knockdown cells. Pretreatment of cells with the protein-tyrosine phosphatase inhibitor pervanadate partially reverses this effect of Grb10 knockdown on receptor phosphorylation, indicating that endogenous Grb10 may block phosphatase access to the activated IGF-I receptor. Marked small interfering RNA knockdown of Grb10 does not result in increased or decreased expression of the related proteins Grb7 or Grb14. As further evidence for Grb10 functional specificity, the recently identified Grb10 interacting GYF proteins are shown to interact specifically with Grb10 and not with Grb7 or Grb14, using yeast two-hybrid assays. We conclude that Grb10 functions as a specific endogenous suppressor of IGF-I-stimulated cell signaling and DNA synthesis. Modulation of the Grb10-IGF-I receptor pathway may represent a mechanism that regulates IGF-I-responsive cell and tissue growth. PMID- 16037383 TI - Estrogen and exercise may enhance beta-cell function and mass via insulin receptor substrate 2 induction in ovariectomized diabetic rats. AB - The prevalence and progression of type 2 diabetes have increased remarkably in postmenopausal women. Although estrogen replacement and exercise have been studied for their effect in modulating insulin sensitivity in the case of insufficient estrogen states, their effects on beta-cell function and mass have not been studied. Ovariectomized (OVX) female rats with 90% pancreatectomy were given a 30% fat diet for 8 wk with a corresponding administration of 17beta estradiol (30 microg/kg body weight) and/or regular exercise. Amelioration of insulin resistance by estrogen replacement or exercise was closely related to body weight reduction. Insulin secretion in first and second phases was lower in OVX during hyperglycemic clamp, which was improved by estrogen replacement and exercise but not by weight reduction induced by restricted diets. Both estrogen replacement and exercise overcame reduced pancreatic beta-cell mass in OVX rats via increased proliferation and decreased apoptosis of beta-cells, but they did not exhibit an additive effect. However, restricted diets did not stimulate beta cell proliferation. Increased beta-cell proliferation was associated with the induction of insulin receptor substrate-2 and pancreatic homeodomain protein-1 via the activation of the cAMP response element binding protein. Estrogen replacement and exercise shared a common pathway, which led to the improvement of beta-cell function and mass, via cAMP response element binding protein activation, explaining the lack of an additive effect with combined treatments. In conclusion, decreased beta-cell mass leading to impaired insulin secretion triggers glucose dysregulation in estrogen insufficiency, regardless of body fat. Regular moderate exercise eliminates the risk factors of contracting diabetes in the postmenopausal state. PMID- 16037384 TI - Resveratrol regulates insulin-like growth factor-II in breast cancer cells. AB - IGF-II is a potent mitogen and inhibitor of apoptosis in breast cancer. Regulation of IGF-II is complex and includes inhibition by tumor suppressors, stimulation by oncogenes, and imprinting and hormonal regulation by estrogens. Resveratrol (RSV) is a phytoestrogen that displays estrogen-like agonistic and antagonistic activity. Recent studies have shown that RSV inhibits the growth of breast cancer cells and may represent a potent agent in chemopreventive therapy. Because 17beta-estradiol regulates IGF-II, we hypothesized that RSV may have a similar effect on IGF-II. The present study was designed to examine whether: 1) RSV modulates IGF-II in breast cancer cells; 2) regulation of IGF-II by RSV is dependent on the ER status; and 3) IGF-II (not IGF-I) mediates RSV effects on breast cancer cells. Treatment of MCF-7 and T47D cells with RSV (10(-6) M) caused stimulation of precursor IGF-II mRNA and protein; this effect was blocked by coincubation with 17beta-estradiol (10(-9) M). Cell growth stimulated by RSV (10( 6) M) was blocked by addition of a blocking IGF-I receptor antibody, or the antiestrogen tamoxifen (10(-7) M). In contrast, RSV treatment (10(-4) M) inhibited IGF-II secretion and cell growth in MCF-7 and T47D cells. No increase in IGF-II levels is seen in estrogen receptor (-) MCF-10 cells, even though cell growth was inhibited by RSV 10(-4) M and precursor IGF-II blocked the inhibitory effect of resveratrol. No change in IGF-I was observed with RSV treatment (10(-6) to 10(-4) M). Our study demonstrates that RSV regulates IGF-II and that IGF-II mediates RSV effect on cell survival and growth in breast cancer cells. PMID- 16037385 TI - Human CTLA4 knock-in mice unravel the quantitative link between tumor immunity and autoimmunity induced by anti-CTLA-4 antibodies. AB - Although results from preclinical studies in animal models have proven the concept for use of anti-cytotoxic T-lymphocyte antigen 4 (CTLA-4) antibodies in cancer immunotherapy, 2 major obstacles have hindered their successful application for human cancer therapy. First, the lack of in vitro correlates of the antitumor effect of the antibodies makes it difficult to screen for the most efficacious antibody by in vitro analysis. Second, significant autoimmune side effects have been observed in a recent clinical trial. In order to address these 2 issues, we have generated human CTLA4 gene knock-in mice and used them to compare a panel of anti-human CTLA-4 antibodies for their ability to induce tumor rejection and autoimmunity. Surprisingly, while all antibodies induced protection against cancer and demonstrated some autoimmune side effects, the antibody that induced the strongest protection also induced the least autoimmune side effects. These results demonstrate that autoimmune disease does not quantitatively correlate with cancer immunity. Our approach may be generally applicable to the development of human therapeutic antibodies. PMID- 16037386 TI - CCR2 is required for CD8-induced graft-versus-host disease. AB - Graft-versus-host disease (GVHD) is a major complication of allogeneic hematopoietic stem cell transplantation (HSCT). Migration of donor-derived T cells into GVHD target organs plays a critical role in the development of GVHD and chemokines and their receptors are important molecules involved in this process. Here, we demonstrate in murine bone marrow transplantation models that the expression of the inflammatory CC chemokine receptor 2 (CCR2) on donor derived CD8+ T cells is relevant for the control of CD8+ T-cell migration and development of GVHD. Recipients of CCR2-deficient (CCR2-/-) CD8+ T cells developed less damage of gut and liver than recipients of wild-type CD8+ T cells, which correlated with a reduction in overall GVHD morbidity and mortality. Assessment of donor CD8+ T-cell target organ infiltration revealed that CCR2-/- CD8+ T cells have an intrinsic migratory defect to the gut and liver. Other causes for the reduction in GVHD could be excluded, as alloreactive proliferation, activation, IFN-gamma production and cytotoxicity of CCR2-/- CD8+ T cells were intact. Interestingly, the graft-versus-tumor effect mediated by CCR2-/- CD8+ T cells was preserved, which suggests that interference with T-cell migration by blockade of CCR2 signaling can separate GVHD from GVT activity. PMID- 16037387 TI - JAK2 mutation 1849G>T is rare in acute leukemias but can be found in CMML, Philadelphia chromosome-negative CML, and megakaryocytic leukemia. AB - An activating 1849G>T mutation of JAK2 (Janus kinase 2) tyrosine kinase was recently described in chronic myeloproliferative disorders (MPDs). Its role in other hematologic neoplasms is unclear. We developed a quantitative pyrosequencing assay and analyzed 374 samples of hematologic neoplasms. The mutation was frequent in polycythemia vera (PV) (86%) and myelofibrosis (95%) but less prevalent in acute myeloid leukemia (AML) with an antecedent PV or myelofibrosis (5 [36%] of 14 patients). JAK2 mutation was also detected in 3 (19%) of 16 patients with Philadelphia-chromosome (Ph)-negative chronic myelogenous leukemia (CML), 2 (18%) of 11 patients with megakaryocytic AML, 7 (13%) of 52 patients with chronic myelomonocytic leukemia, and 1 (1%) of 68 patients with myelodysplastic syndromes. No mutation was found in Ph(+)CML (99 patients), AML M0-M6 (28 patients), or acute lymphoblastic leukemia (20 patients). We conclude that the JAK2 1849G>T mutation is common in Ph(-) MPD but not critical for transformation to the acute phase of these diseases and that it is generally rare in aggressive leukemias. PMID- 16037388 TI - Induction of HM1.24 peptide-specific cytotoxic T lymphocytes by using peripheral blood stem-cell harvests in patients with multiple myeloma. AB - HM1.24 antigen is preferentially overexpressed in multiple myeloma (MM) cells but not in normal cells. To explore the potential of HM1.24 as a target for cellular immunotherapy, we selected 4 HM1.24-derived peptides that possess binding motifs for HLA-A2 or HLA-A24 by using 2 computer-based algorithms. The ability of these peptides to generate cytotoxic T lymphocytes (CTLs) was examined in 20 healthy donors and 6 patients with MM by a reverse immunologic approach. Dendritic cells (DCs) were induced from peripheral-blood mononuclear cells of healthy donors or peripheral-blood stem-cell (PBSC) harvests from patients with MM, and autologous CD8(+) T cells were stimulated with HM1.24 peptide-pulsed DCs. Both interferon gamma-producing and cytotoxic responses were observed after stimulation with either HM1.24-126 or HM1.24-165 peptides in HLA-A2 or HLA-A24 individuals. The peptide-specific recognition of these CTLs was further confirmed by tetramer assay and cold target inhibition assay. Importantly, HM1.24-specific CTLs were also induced from PBSC harvests from patients with MM and these CTLs were able to kill MM cells in an HLA-restricted manner. These results indicate the existence of functional DCs and HM1.24-specific CTL precursors within PBSC harvests and provide the basis for cellular immunotherapy in combination with autologous PBSC transplantation in MM. PMID- 16037390 TI - Excessive fluid accumulation during stem cell mobilization: a novel prognostic factor of first-year survival after stem cell transplantation in AL amyloidosis patients. AB - High-dose melphalan followed by stem cell transplantation (HDM-SCT) has become the treatment of choice for patients with immunoglobulin light chain amyloidosis (AL). Unfortunately, treatment mortality can be excessive in certain subpopulations. We have noted that patients who gained more than 2% body weight during mobilization had a poorer outcome following HDM-SCT. Excluding 2 patients for lack of weight record and denial of consent, 126 patients between July 1997 and June 2003 were retrospectively studied. Weight increased more than 2.0% during mobilization in 51.6% of the patients. Patients who accumulated more than 2.0% tended to have more proteinuria, more organs involved, lower serum albumin, more diuretic use, and dosage adjustment during mobilization. First-year mortality was significantly higher in those with more than 2% weight gain (33.9% versus 9.8%, P = .002). Multivariate analysis showed weight gain, glomerular filtration rate, and septal thickness to be independent predictors of first-year mortality. The increase in mortality was noted even after the excess weight was diuresed prior to conditioning. The impact on mortality dissipated after the first year. Weight gain during mobilization appears to be a new marker of adverse outcome following HDM-SCT. Better prognostication may improve the treatment mortality rate of these patients. PMID- 16037389 TI - Neutrophil stimulation with Mycobacterium bovis bacillus Calmette-Guerin (BCG) results in the release of functional soluble TRAIL/Apo-2L. AB - Mycobacterium bovis bacillus Calmette-Guerin (BCG) has been used to treat bladder cancer for almost 30 years; however, the effector mechanism of the BCG-induced antitumor response remains enigmatic. Most BCG research has focused on the mononuclear-cell infiltrate, but growing evidence supports a role for neutrophils in the antitumor response. Previously, we demonstrated increased urinary tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL/Apo-2L) levels from BCG-responsive patients compared to nonresponders. Interestingly, neutrophils isolated from the urine expressed TRAIL/Apo-2L, leading us to investigate the neutrophil response to BCG. BCG-stimulated neutrophils expressed surface-bound and released functional soluble TRAIL/Apo-2L. Whereas neither interferon alpha (IFN-alpha) nor IFN-gamma directly induced TRAIL/Apo2L expression by neutrophils, IFN-alpha did stimulate TRAIL gene transcription, and IFN-primed neutrophils contained and released more TRAIL/Apo-2L after BCG stimulation than did unprimed neutrophils. In unstimulated neutrophils TRAIL/Apo 2L was present predominantly in the azurophilic granules and plasma-membrane enriched/secretory-granule fraction. Finally, we observed that killed BCG, Toll like receptor 2 (TLR2) and TLR4 agonists, and an M tuberculosis cell-wall fraction were each capable of inducing the release of soluble TRAIL/Apo-2L from neutrophils. These results further characterize the potential role neutrophils may play in initiating the antitumor response described with BCG treatment for superficial bladder cancer. PMID- 16037391 TI - Functional aberrant expression of CCR2 receptor on chronically activated NK cells in patients with TAP-2 deficiency. AB - Chemokines play a pivotal role in homeostatic and inflammatory migration of naive and activated natural killer (NK) subsets. Recent studies have shown that aberrant chemokine receptor expression on certain immune cells underlies the pathogenesis of clinical conditions in which recruitment of such cells is altered. Progressive accumulation of activated NK cells, subsequently resulting in the formation of chronic granulomatous lesions in the respiratory tract and the skin, has been described in a number of patients with transporter associated with antigen processing 2 (TAP-2) deficiency in the later stages of disease. Therefore, the goal of the present study was to elucidate whether the dysregulation of chemoattracting receptor expression on NK cells could explain abnormal navigation of these cells in TAP-2 deficiency. High-throughput proteomic comparison, followed by verification with flow cytometry, revealed that chronically activated NK cells derived from 3 newly identified patients with TAP 2 deficiency consistently expressed aberrant levels of CC chemokine receptor 2 (CCR2) chemokine receptor in vitro and in vivo. This expression pattern translated into specific responsiveness of chronically activated NK cells derived from patients with TAP-2 deficiency to multiple ligands of CCR2. Moreover, the in vivo elevated levels of interleukin-2 (IL-2) and monocyte chemoattractant protein 1 (MCP-1) detected in serum and bronchoalveolar lavage samples derived from these patients highlight the potential involvement of the CCR2 pathway in aberrant NK cell retention at chronic inflammatory sites. PMID- 16037392 TI - Platelet aggregation induces platelet aggregate stability via SLAM family receptor signaling. AB - Platelet aggregation is a dynamic entity, capable of directing its own growth and stability via the activation of signaling cascades that lead to the expression and secretion of various secondary agonists. Here we show that the signaling pathways triggered during platelet aggregation include an intrinsic pro thrombotic activity mediated by 2 homophilic adhesion molecules, CD84 and CD150 (SLAM [signaling lymphocyte activation molecule]), which are tyrosine phosphorylated in a platelet aggregation-dependent fashion. The 2 CD84/SLAM adapter proteins, SAP (SLAM-associated protein) and EAT-2 (EWS-activated transcript-2), were found in platelets; only SAP, however, was found to immunoprecipitate with tyrosine-phosphorylated SLAM. The immobilized extracellular domain of CD84 promoted microaggregate formation, while SAP deficient platelets demonstrated defective spreading on immobilized CD84, demonstrating a functional role in platelets for SLAM family interactions. Finally, analysis of SLAM-deficient mice revealed an overall defect in platelet aggregation in vitro and a delayed arterial thrombotic process in vivo. The data indicate that signaling of the adhesion molecules in the SLAM family, activated by proximity during aggregation, further stabilize platelet-platelet interactions in thrombosis. PMID- 16037393 TI - Distinct roles for the NF-kappaB1 and c-Rel transcription factors in the differentiation and survival of plasmacytoid and conventional dendritic cells activated by TLR-9 signals. AB - Reticuloendotheliosis viral oncogene homolog/nuclear factor of kappa light polypeptide gene enhancer in B cells 1 (Rel/NF-kappaB) activation is a ubiquitous outcome of engaging Toll-like receptors (TLRs), yet the cell-type-specific functions of this pathway in response to particular microbial signals remain poorly defined. Here we show that NF-kappaB1 and C-Rel, Rel/NF-kappaB proteins induced in conventional dendritic cells (cDCs) and plasmacytoid dendritic cells (pDCs) by cytosine-phosphate-guanosine (CpG) DNA, a TLR-9 ligand, serve markedly different functions in these DC subsets. With the exception of impaired interleukin-12 (IL-12) production, cultured Nfkb1(-/-)C-Rel(-/-) cDCs responded relatively normally to CpG DNA. In contrast, CpG-treated Nfkb1(-/-)C-Rel(-/-) pDCs, which were still able to produce type I interferon and regulated on activation normal T-cell expressed and secreted (RANTES), but not IL-6 or IL-12, failed to acquire an activated dendritic phenotype and underwent apoptosis. Although the TLR-9-mediated death of Nfkb1(-/-)C-Rel(-/-) pDCs, which coincided with a failure to up-regulate the prosurvival proteins B-cell lymphoma apoptosis regulator xL (Bcl-x(L)) and A1, was blocked by Bcl-2 transgene expression, this inhibition of apoptosis still failed to rescue the differentiation defects. This indicated that these NF-kappaB transcription factors independently regulate TLR-9 mediated pDC morphogenesis and survival. Collectively, these findings establish that NF-kappaB1 and c-Rel, while largely dispensable for TLR-9-induced cDC activation, are critical for regulating differentiation and survival programs during pDC activation. PMID- 16037394 TI - G-CSF potently inhibits osteoblast activity and CXCL12 mRNA expression in the bone marrow. AB - Accumulating evidence indicates that interaction of stromal cell-derived factor 1 (SDF-1/CXCL12 [CXC motif, ligand 12]) with its cognate receptor, CXCR4 (CXC motif, receptor 4), generates signals that regulate hematopoietic progenitor cell (HPC) trafficking in the bone marrow. During granulocyte colony-stimulating factor (G-CSF)-induced HPC mobilization, CXCL12 protein expression in the bone marrow decreases. Herein, we show that in a series of transgenic mice carrying targeted mutations of their G-CSF receptor and displaying markedly different G CSF-induced HPC mobilization responses, the decrease in bone marrow CXCL12 protein expression closely correlates with the degree of HPC mobilization. G-CSF treatment induced a decrease in bone marrow CXCL12 mRNA that closely mirrored the fall in CXCL12 protein. Cell sorting experiments showed that osteoblasts and to a lesser degree endothelial cells are the major sources of CXCL12 production in the bone marrow. Interestingly, osteoblast activity, as measured by histomorphometry and osteocalcin expression, is strongly down-regulated during G-CSF treatment. However, the G-CSF receptor is not expressed on osteoblasts; accordingly, G-CSF had no direct effect on osteoblast function. Collectively, these data suggest a model in which G-CSF, through an indirect mechanism, potently inhibits osteoblast activity resulting in decreased CXCL12 expression in the bone marrow. The consequent attenuation of CXCR4 signaling ultimately leads to HPC mobilization. PMID- 16037396 TI - Inhibition of inducible nitric oxide synthase attenuates platelet adhesion in subpleural arterioles caused by lung ischemia-reperfusion in rabbits. AB - Oxidative stress, induced by lung ischemia-reperfusion, leads to platelet and leukocyte activation and may contribute to decreased alveolar perfusion by platelet adhesion to the arteriolar wall. We investigated the hypothesis that ischemia-reperfusion injury increases inducible nitric oxide synthase (iNOS) activity and subsequent generation of reactive nitrogen species with P-selectin dependent platelet-endothelial interactions and vasoconstriction during lung reperfusion. Subpleural arterioles, labeled platelets, and leukocytes were examined in anesthetized, open-chest rabbits by intravital fluorescence microscopy. Ischemia was caused by reversible occlusion of the right pulmonary artery for 1 or 2 h (1IR and 2IR groups). During 2 h of reperfusion, postischemic platelet rolling and adhesion were independent from leukocyte-arteriolar wall interactions and correlated with pulmonary arteriolar constriction in proportion to the length of ischemia. In rabbits treated with an iNOS inhibitor (1400W) before occlusion (2IR + 1400W group), platelet-arteriolar wall interactions and vasoconstriction were prevented. iNOS expression and activity in ischemic lung tissue were markedly greater than control and also were proportional to ischemia duration. NOS activity, immunochemically detected P-selectin, and nitrotyrosine expression in ischemic lung tissue from animals subjected to ischemia reperfusion, as well as the plasma level of soluble P-selectin, were significantly higher than in nonischemic lungs and were inhibited by pretreatment with 1400W. These results show that platelet adhesion and arteriolar constriction during early reperfusion in the ventilated lung can result from increased iNOS activity and is highly correlated with reactive nitrogen species and P-selectin expression. PMID- 16037395 TI - Repeat exercise normalizes the gas-exchange impairment induced by a previous exercise bout in asthmatic subjects. AB - Twenty-one subjects with asthma underwent treadmill exercise to exhaustion at a workload that elicited approximately 90% of each subject's maximal O2 uptake (EX1). After EX1, 12 subjects experienced significant exercise-induced bronchospasm [(EIB+), %decrease in forced expiratory volume in 1.0 s = -24.0 +/- 11.5%; pulmonary resistance at rest vs. postexercise = 3.2 +/- 1.5 vs. 8.1 +/- 4.5 cmH2O.l(-1).s(-1)] and nine did not (EIB-). The alveolar-to-arterial Po2 difference (A-aDo2) was widened from rest (9.1 +/- 6.7 Torr) to 23.1 +/- 10.4 and 18.1 +/- 9.1 Torr at 35 min after EX1 in subjects with and without EIB, respectively (P < 0.05). Arterial Po2 (PaO2) was reduced in both groups during recovery (EIB+, -16.0 +/- -13.0 Torr vs. baseline; EIB-, -11.0 +/- 9.4 Torr vs. baseline, P < or = 0.05). Forty minutes after EX1, a second exercise bout was completed at maximal O2 uptake. During the second exercise bout, pulmonary resistance decreased to baseline levels in the EIB+ group and the A-aDo2 and PaO2 returned to match the values seen during EX1 in both groups. Sputum histamine (34.6 +/- 25.9 vs. 61.2 +/- 42.0 ng/ml, pre- vs. postexercise) and urinary 9alpha,11beta-prostaglandin F2 (74.5 +/- 38.6 vs. 164.6 +/- 84.2 ng/mmol creatinine, pre- vs. postexercise) were increased after exercise only in the EIB+ group (P < 0.05), and postexercise sputum histamine was significantly correlated with the exercise PaO2 and A-aDo2 in the EIB+ subjects. Thus exercise causes gas exchange impairment during the postexercise period in asthmatic subjects independent of decreases in forced expiratory flow rates after the exercise; however, a subsequent exercise bout normalizes this impairment secondary in part to a fast acting, robust exercise-induced bronchodilatory response. PMID- 16037397 TI - Sprint training improves postischemic, left ventricular diastolic performance. AB - We examined the effects of sprint training on left ventricular diastolic stiffness during normoxia and after ischemia-reperfusion (I/R). Thirty-seven, male Sprague-Dawley rats, weighing 150-175 g at the initiation of the experiment, were randomly assigned to a sedentary, control group (n = 20) or to a high intensity, sprint-trained group (n = 17). Animals were trained 5 days/wk on a motor-driven treadmill for 6 wk. High-intensity sprint training consisted of running five 1-min sprints at 75 m/min, 15% grade, interspersed with 1-min active recovery runs at a speed of 20 m/min, 15% grade. Langendorff-derived isolated heart performance was measured before and after 20 min of no-flow ischemia followed by 30 min of reperfusion. Isolated myocytes were harvested from a subset of postischemic hearts. Sprint training reduced Langendorff-derived LV chamber stiffness (P < 0.05) and induced a rightward shift in the LV pressure-volume relationship during both normoxic perfusion and after I/R. LV developed pressure after I/R was also better preserved in hearts obtained from sprint-trained animals (P < 0.05), a result that is in part related to a lower postischemic LV chamber stiffness in sprint-trained hearts. The putative impact of sprint training on postischemic LV chamber stiffness was masked by glycolytic inhibition with iodoacetate, suggesting that glycolysis was involved in the better postischemic recovery observed in sprint-trained hearts. There was a tendency for enhanced postischemic cardiomyocyte shortening in sprint-trained cardiomyocytes compared with control. The rate of myocyte relaxation, i.e., time for 50% relaxation of the Ca(2+) transient amplitude, was similar between groups. These data suggest that additional mechanisms unrelated to Ca(2+) were involved in sprint-induced protection from ischemia-reperfusion-induced LV diastolic dysfunction. PMID- 16037398 TI - Kinetics of O2 uptake, leg blood flow, and muscle deoxygenation are slowed in the upper compared with lower region of the moderate-intensity exercise domain. AB - Six male subjects [23 yr (SD 4)] performed repetitions (6-8) of two-legged, moderate-intensity, knee-extension exercise during two separate protocols that included step transitions from 3 W to 90% estimated lactate threshold (thetaL) performed as a single step (S3) and in two equal steps (S1, 3 W to approximately 45% thetaL; S2, approximately 45% thetaL to approximately 90% thetaL). The time constants (tau) of pulmonary oxygen uptake (Vo2), leg blood flow (LBF), heart rate (HR), and muscle deoxygenation (HHb) were greater (P < 0.05) in S2 (tauVo2, approximately 52 s; tauLBF, approximately 39 s; tauHR, approximately 42 s; tauHHb, approximately 33 s) compared with S1 (tauVo2, approximately 24 s; tauLBF, approximately 21 s; tauHR, approximately 21 s; tauHHb, approximately 16 s), while the delay before an increase in HHb was reduced (P < 0.05) in S2 (approximately 14 s) compared with S1 (approximately 20 s). The Vo2 and HHb amplitudes were greater (P < 0.05) in S2 compared with S1, whereas the LBF amplitude was similar in S2 and S1. Thus the slowed Vo2 response in S2 compared with S1 is consistent with a mechanism whereby Vo2 kinetics is limited, in part, by a slowed adaptation of blood flow and/or O2 transport when exercise was initiated from a baseline of moderate-intensity exercise. PMID- 16037399 TI - Kidneys extract BNP and NT-proBNP in healthy young men. AB - Renal metabolism of the cardiac marker NH2-terminal-pro-brain natriuretic peptide (NT-proBNP) has been suggested. Therefore, we determined the renal extraction ratios of NT-proBNP and its bioactive coproduct brain natriuretic peptide (BNP) at rest and during exercise. In addition, the cerebral ratios were evaluated. Ten young healthy men were investigated at baseline, during moderate cycle exercise (heart rate: 140, Borg scale: 14-15), and in the recovery with BNP and NT-proBNP measured from the brachial artery and the jugular and renal veins, and the renal and cerebral extraction ratios (Ext-Ren and Ext-Cer, respectively) were calculated. Cardiac output, stroke volume, heart rate, mean arterial pressures, and estimated glomerular filtration were determined. BNP and NT-proBNP were extracted by the kidneys but not by the brain. We observed no effect of exercise. The mean values (+/- SE) of Ext-Ren of NT-proBNP were similar (0.19 +/- 0.05, 0.21 +/- 0.06, and 0.12 +/- 0.03, respectively) during the three sessions (P > 0.05). Also the Ext-Ren of BNP were similar (0.18 +/- 0.07, 0.15 +/- 0.11, and 0.14 +/- 0.06, respectively; P > 0.05). There were no significant differences between Ext-Ren of BNP and NT-proBNP during the three sessions (P > 0.05). The Ext-Cer of both peptides varied insignificantly between -0.21 +/- 0.15 and 0.11 +/- 0.08. The renal extraction ratio of both BNP and NT-proBNP is approximately 0.15-0.20. There is no cerebral extraction, and short-term moderate exercise does not affect these values. Our findings suggest that the kidneys extract BNP and NT proBNP to a similar extent in healthy young men. PMID- 16037400 TI - Exercise hyperemia and vasoconstrictor responses in humans with cystic fibrosis. AB - ATP released from circulating erythrocytes is a potential signal regulating muscle blood flow during exercise (exercise hyperemia), and intravascular ATP appears to blunt sympathetic vasoconstriction during exercise. Erythrocytes from patients with cystic fibrosis (CF) do not release ATP. The goal of the present study was to determine whether increases in forearm blood flow during exercise are blunted in CF patients and whether CF patients exhibit greater vasoconstrictor responsiveness during exercise. Nine control subjects and 10 CF patients who were free of other disease complications (approximately 96% O2 saturation) performed incremental rhythmic forearm exercise at 5, 10, and 15% of maximum handgrip strength for 21 min (7 min at each workload). We used a cold pressor test to evoke sympathetic vasoconstriction under resting conditions and at each exercise workload. As a control, subjects performed a second exercise bout without the cold pressor test. Continuous brachial artery blood velocity was monitored beat-to-beat, and vessel diameter was assessed by Doppler ultrasound. Artery diameter, as well as blood pressure, heart rate, and O2 saturation, was measured at steady-state exercise and at 1 min into the cold pressor stimulus. Blood pressure and heart rate responses to the forearm exercise and each cold pressor test were similar in both groups (P > 0.05). Contrary to our hypothesis, forearm blood flow (P = 0.91) and forearm vascular conductance (P = 0.82) were similar at rest and at each level of exercise between CF patients and controls. Additionally, there was no difference in the degree of sympathetic vasoconstriction between groups at rest and at each level of exercise (P = 0.22). Our results suggest that ATP released from the deformation of erythrocytes is not an obligatory signal for exercise hyperemia in human skeletal muscle. PMID- 16037402 TI - Nonthermoregulatory control of cutaneous vascular conductance and sweating during recovery from dynamic exercise in women. AB - The purpose of the study was to examine the effect of 1) active (loadless pedaling), 2) passive (assisted pedaling), and 3) inactive (motionless) recovery modes on mean arterial pressure (MAP), cutaneous vascular conductance (CVC), and sweat rate during recovery after 15 min of dynamic exercise in women. It was hypothesized that an active recovery mode would be most effective in attenuating the fall in MAP, CVC, and sweating during exercise recovery. Ten female subjects performed 15 min of cycle ergometer exercise at 70% of their predetermined peak oxygen consumption followed by 20 min of 1) active, 2) passive, or 3) inactive recovery. Mean skin temperature (Tsk), esophageal temperature (Tes), skin blood flow, sweating, cardiac output (CO), stroke volume (SV), heart rate (HR), total peripheral resistance (TPR), and MAP were recorded at baseline, end exercise, and 2, 5, 8, 12, 15, and 20 min postexercise. Cutaneous vascular conductance (CVC) was calculated as the ratio of laser-Doppler blood flow to MAP. In the active recovery mode, CVC, sweat rate, MAP, CO, and SV remained elevated over inactive values (P < 0.05). The passive mode was equally as effective as the active mode in maintaining MAP. Sweat rate was different among all modes after 12 min of recovery (P < 0.05). TPR during active recovery remained significantly lower than during recovery in the inactive mode (P < 0.05). No differences in either Tes or Tsk were observed among conditions. The results indicate that CVC can be modulated by central command and possibly cardiopulmonary baroreceptors in women. However, differences in sweat rate may be influenced by factors such as central command, mechanoreceptor stimulation, or cardiopulmonary baroreceptors. PMID- 16037401 TI - Chronic intermittent hypoxia upregulates genes of lipid biosynthesis in obese mice. AB - Obstructive sleep apnea (OSA), a condition tightly linked to obesity, leads to chronic intermittent hypoxia (CIH) during sleep. There is emerging evidence that OSA is independently associated with insulin resistance and fatty liver disease, suggesting that OSA may affect hepatic lipid metabolism. To test this hypothesis, leptin-deficient obese (ob/ob) mice were exposed to CIH during the light phase (9 AM-9 PM) for 12 wk. Liver lipid content and gene expression profile in the liver (Affymetrix 430 GeneChip with real-time PCR validation) were determined on completion of the exposure. CIH caused a 30% increase in triglyceride and phospholipid liver content (P < 0.05), whereas liver cholesterol content was unchanged. Gene expression analysis showed that CIH upregulated multiple genes controlling 1) cholesterol and fatty acid biosynthesis [malic enzyme and acetyl coenzyme A (CoA) synthetase], 2) predominantly fatty acid biosynthesis (acetyl CoA carboxylase and stearoyl-CoA desaturases 1 and 2), and 3) triglyceride and phospholipid biosynthesis (mitochondrial glycerol-3-phosphate acyltransferase). A majority of overexpressed genes were transcriptionally regulated by sterol regulatory element-binding protein (SREBP) 1, a master regulator of lipogenesis. A 2.8-fold increase in SREBP-1 gene expression in CIH was confirmed by real-time PCR (P = 0.001). Expression of major genes of cholesterol biosynthesis, SREBP-2 and 3-hydroxy-3-methylglutaryl-CoA reductase, was unchanged. In conclusion, we have shown that CIH may exacerbate preexisting fatty liver of obesity via upregulation of the pathways of lipid biosynthesis in the liver. PMID- 16037403 TI - An in vivo microanalytical technique for measuring the local biochemical milieu of human skeletal muscle. AB - Myofascial pain associated with myofascial trigger points (MTrPs) is a common cause of nonarticular musculoskeletal pain. Although the presence of MTrPs can be determined by soft tissue palpation, little is known about the mechanisms and biochemical milieu associated with persistent muscle pain. A microanalytical system was developed to measure the in vivo biochemical milieu of muscle in near real time at the subnanogram level of concentration. The system includes a microdialysis needle capable of continuously collecting extremely small samples (approximately 0.5 microl) of physiological saline after exposure to the internal tissue milieu across a 105-microm-thick semi-permeable membrane. This membrane is positioned 200 microm from the tip of the needle and permits solutes of <75 kDa to diffuse across it. Three subjects were selected from each of three groups (total 9 subjects): normal (no neck pain, no MTrP); latent (no neck pain, MTrP present); active (neck pain, MTrP present). The microdialysis needle was inserted in a standardized location in the upper trapezius muscle. Due to the extremely small sample size collected by the microdialysis system, an established microanalytical laboratory, employing immunoaffinity capillary electrophoresis and capillary electrochromatography, performed analysis of selected analytes. Concentrations of protons, bradykinin, calcitonin gene-related peptide, substance P, tumor necrosis factor-alpha, interleukin-1beta, serotonin, and norepinephrine were found to be significantly higher in the active group than either of the other two groups (P < 0.01). pH was significantly lower in the active group than the other two groups (P < 0.03). In conclusion, the described microanalytical technique enables continuous sampling of extremely small quantities of substances directly from soft tissue, with minimal system perturbation and without harmful effects on subjects. The measured levels of analytes can be used to distinguish clinically distinct groups. PMID- 16037404 TI - Adrenergic receptors mediate stress-induced elevations in extracellular Hsp72. AB - Heat-shock protein concentrations in the blood increase after exposure to a variety of stressors, including trauma and psychological stress. Although the physiological function of extracellular heat shock protein remains controversial, there is evidence that extracellular heat shock protein 72 (Hsp72) can facilitate immunologic responses. The signal(s) that mediate(s) the in vivo elevation of extracellular Hsp72 in the blood after stressor exposure remain(s) unknown. Here we report that Hsp72 increases in the circulation via an alpha1-adrenergic receptor-mediated signaling pathway. Activation of alpha1-adrenoceptors results in a rapid increase in circulating Hsp72, and blockade of alpha1-adrenoceptors prevents the stress-induced rise in circulating Hsp72. Furthermore, our studies exclude a role for beta-adrenoceptors, glucocorticoids, and ACTH in mediating stress-induced elevations in circulating extracellular Hsp72. Understanding the signals involved in elevating extracellular Hsp72 could facilitate the use of extracellular Hsp72 to bolster immunity and perhaps prevent exacerbation of inflammatory diseases during stress. PMID- 16037405 TI - GABAB-receptor-mediated suppression of sympathetic outflow from the spinal cord of neonatal rats. AB - Using a splanchnic nerve-spinal cord preparation in vitro that could spontaneously generate sympathetic nerve discharge (SND), we investigated the roles of intraspinal GABA(B) receptors in the regulation of SND. Despite an age dependent difference in sensitivity, bath applications of baclofen (Bac; GABA(B) receptor agonist) consistently reduced SND in a concentration-dependent manner. The drug specificity of Bac in activation of GABA(B) receptors was verified by application of its antagonist saclofen (Sac) or CGP-46381 (CGP). Sac or CGP alone did not change SND. However, in the presence of Sac or CGP, the effects of Bac on SND inhibition were reversibly attenuated. The splanchnic sympathetic preganglionic neuron (SPN) was recorded by blind whole cell, patch-clamp techniques. We examined Bac effects on electrical membrane properties of SPNs. Applications of Bac reduced excitatory synaptic events, induced membrane hyperpolarizations, and inhibited SPN firing. In the presence of 12 mM Mg2+ or 0.5 microM TTX to block Ca2+- or action potential-dependent synaptic transmissions, applications of Bac induced an outward baseline current that reversed at -29 +/- 6 mV. Because the K+ equilibrium potential in our experimental conditions was -100 mV, the Bac-induced currents could not simply be attributed to an alteration of K+ conductance. On the other hand, applications of Bac to Cs+-loaded SPNs reduced Cd2+-sensitive and high-voltage-activated inward currents, indicating an inhibition of voltage-gated Ca2+ currents. Our results suggest that the activation of intraspinal GABA(B) receptors suppresses SND via a mixture of ion events that may link to a change in Ca2+ conductance. PMID- 16037406 TI - Impact of glutamine supplementation on glucose homeostasis during and after exercise. AB - The interaction of glutamine availability and glucose homeostasis during and after exercise was investigated, measuring whole body glucose kinetics with [3 3H]glucose and net organ balances of glucose and amino acids (AA) during basal, exercise, and postexercise hyperinsulinemic-euglycemic clamp periods in six multicatheterized dogs. Dogs were studied twice in random treatment order: once with glutamine (12 micromol.kg(-1).min(-1); Gln) and once with saline (Con) infused intravenously during and after exercise. Plasma glucose fell by 7 mg/dl with exercise in Con (P < 0.05), but it did not fall with Gln. Gln further stimulated whole body glucose production and utilization an additional 24% above a normal exercise response (P < 0.05). Net hepatic uptake of glutamine and alanine was greater with Gln than Con during exercise (P < 0.05). Net hepatic glucose output was increased sevenfold during exercise with Gln (P < 0.05) but not with Con. Net hindlimb glucose uptake was increased similarly during exercise in both groups (P < 0.05). During the postexercise hyperinsulinemic-euglycemic period, glucose production decreased to near zero with Con, but it did not decrease below basal levels with Gln. Gln increased glucose utilization by 16% compared with Con after exercise (P < 0.05). Furthermore, net hindlimb glucose uptake in the postexercise period was increased approximately twofold vs. basal with Gln (P < 0.05) but not with Con. Net hepatic uptake of glutamine during the postexercise period was threefold greater for Gln than Con (P < 0.05). In conclusion, glutamine availability modulates glucose homeostasis during and after exercise, which may have implications for postexercise recovery. PMID- 16037407 TI - Clonidine induces nitric oxide- and prostaglandin-mediated vasodilation in healthy human skin. AB - Sustained sympathetic activation not only leads to vasoconstriction but also might induce paradox vasodilation. This study was performed to explore whether and how alpha(2)-receptor stimulation mediates this vasodilation. We investigated 11 healthy subjects in 33 dermal microdialysis (MD) sessions. After nerve trunk blockade, MD fibers were inserted and perfused with physiological saline until skin trauma-related vasodilation subsided. Thereafter, fibers were perfused with either clonidine solutions (10(-3), 5 x 10(-4), 10(-4) mol/l), N(G)-monomethyl-l arginine (L-NMMA; nitric oxide synthase blocker), acetylsalicylic acid (ASA; cyclooxygenase blocker), or combinations of these. Laser-Doppler scanning of the investigated skin revealed that clonidine not only induces vasoconstriction but subsequently also vasodilation with higher concentrations (P < 0.001). In contrast, both L-NMMA and ASA induced vasoconstriction (P < 0.001). By coapplication of 10(-3) mol/l clonidine with L-NMMA or ASA, vasodilation was partially prevented (P < 0.001). Our results demonstrate that sustained alpha(2) receptor stimulation induces vasodilation in a dose-dependent way, which is mediated by nitric oxide and prostaglandin mechanisms in human skin. PMID- 16037409 TI - Macrophages in the murine pancreas and their involvement in fetal endocrine development in vitro. AB - Macrophages are a heterogeneous population of cells that belong to the mononuclear phagocyte system. They play an important role in tissue homeostasis and remodeling and are also potent immune regulators. Pancreatic macrophages are critically involved in the development and pathogenesis of autoimmune diabetes. To elucidate the ontogeny of pancreatic macrophages, we characterized in this study the macrophages present in the adult and developing fetal pancreas of normal mice. We additionally examined the presence of local macrophage precursors and the involvement of macrophages in the growth of endocrine tissue in the fetal pancreas. We identified two phenotypically distinct macrophage subsets in the adult pancreas. The majority of macrophages was CD45(+)ER-MP23(+)MOMA-1(+). Under noninflammatory conditions, only a minority ( approximately 5%) of the pancreatic macrophages additionally expressed the macrophage marker F4/80. In contrast, in the fetal pancreas, phenotypically, mature macrophages were identified exclusively by their expression of F4/80 and lacked detectable staining with ER MP23 and MOMA-1 antibodies. In fetal pancreas organ cultures, we could show that macrophages develop from pre-existing precursors, which are present in the fetal pancreas at embryonic age 12.5. Moreover, the number of macrophages increased significantly when macrophage-colony stimulating factor was added to these cultures. It is important that this increase of F4/80-positive cells was paralleled by an increase in the number of insulin-producing cells, suggesting that macrophages support the growth of these endocrine cells. PMID- 16037408 TI - A novel role for Notch ligand Delta-1 as a regulator of human Langerhans cell development from blood monocytes. AB - Human Langerhans cells (LCs) are of hematopoietic origin, but cytokine regulation of their development is not fully understood. Notch ligand Delta-1 is expressed in a proportion of the skin. Granulocyte-macrophage colony-stimulating factor (GM CSF) and transforming growth factor-beta1 (TGF-beta1) are also secreted in the skin. We report here that Delta-1, in concert with GM-CSF and TGF-beta1, induces the differentiation of human CD14(+) blood monocytes into cells that express LC markers: CD1a, Langerin, cutaneous lymphocyte-associated antigen, CC chemokine receptor 6, E-cadherin, and Birbeck granules. The resulting cells display phagocytic activity and chemotaxis to macrophage inflammatory protein-1alpha (MIP 1alpha). In response to CD40 ligand and tumor necrosis factor alpha, the cells acquire a mature phenotype of dendritic cells that is characterized by up regulation of human leukocyte antigen (HLA)-ABC, HLA-DR, CD80, CD86, CD40, and CD54 and appearance of CD83. These cells in turn show chemotaxis toward MIP-1beta and elicit activation of CD8(+) T cells and T helper cell type 1 polarization of CD4(+) T cells. Thus, blood monocytes can give rise to LCs upon exposure to the skin cytokine environment consisting of Delta-1, GM-CSF, and TGF-beta1, which may be, in part, relevant to the development of human epidermal LCs. Our results extend the functional scope of Notch ligand delta-1 in human hematopoiesis. PMID- 16037410 TI - Dendritic cells differentiated in the presence of IFN-{beta} and IL-3 are potent inducers of an antigen-specific CD8+ T cell response. AB - Dendritic cells (DC) are professional antigen-presenting cells that are used in vaccine approaches to cancer. Classically, mature monocyte-derived DC are generated in vitro in the presence of interleukin (IL)-4, granulocyte macrophage colony stimulating factor, and inflammatory cytokines (G4-DC). Recently, it has been described that DC can also be generated in the presence of IL-3 and interferon (IFN)-beta and that these DC are efficiently matured using polyriboinosinic polyribocytidylic acid (I3-DC). In this study, a series of in vitro experiments was performed to compare side-by-side I3-DC and G4-DC as vaccine adjuvants. Phenotypic characterization of the DC revealed differences in the expression of the monocyte marker CD14 and the maturation marker CD83. Low expression of CD14 and high expression of CD83 characterized G4-DC, whereas I3-DC displayed intermediate expression of CD14 and CD83. Both types of DC were as potent in the induction of allogeneic T cell proliferation. Upon CD40 ligation, G4-DC produced lower amounts of IFN-alpha and pulmonary and activation-regulated chemokine, similar amounts of IL-6, macrophage-inflammatory protein (MIP)-1alpha, and MIP-1beta, and higher amounts of IL-12 p70, tumor necrosis factor alpha, and MIP-3beta than I3-DC. We further evaluated whether the DC could be frozen/thawed without loss of cell number, viability, phenotype, and function. After freezing/thawing, 56.0% +/- 9.0% of I3-DC and 77.0% +/- 3.0% of G4-DC (n=9) were recovered as viable cells, displaying the same phenotype as their fresh counterparts. Finally, in vitro stimulations showed that fresh and frozen peptide loaded I3-DC are more potent inducers of Melan-A-specific CD8(+) T cell responses than G4-DC. The antigen-specific T cells were functional as shown in cytotoxicity and IFN-gamma secretion assay. PMID- 16037411 TI - Dopamine contamination of infused tyramine. PMID- 16037412 TI - Serum parathyroid hormone concentrations are increased in women with polycystic ovary syndrome. AB - BACKGROUND: The present study was designed to investigate the effects of polycystic ovary syndrome (PCOS) and of obesity on serum parathyroid hormone (RhoTauEta), 25-hydroxyvitamin D (25-OH-vitamin D), and 1,25-dihydroxyvitamin D [1,25-(OH)2-vitamin D] concentrations and the possible associations of the above calciotropic hormones with the hormonal and metabolic characteristics of the syndrome. METHODS: We studied 58 obese [body mass index (BMI)>30 kg/m2] women with PCOS, 64 overweight (BMI, 25-30 kg/m2) women with the syndrome, 169 normal weight (BMI<25 kg/m2) women with PCOS, 29 obese controls (ovulatory women without clinical or biochemical hyperandrogenemia), 14 overweight controls, and 70 normal weight controls. Blood samples were collected (at 0900 after an overnight fast) between the 3rd and 6th days of a menstrual cycle in the control groups and during a spontaneous bleeding episode in the PCOS groups. Circulating concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin (PRL), testosterone, Delta4-androstenedione, 17alpha hydroxyprogesterone, sex-hormone-binding globulin (SHBG), insulin, glucose, PTH, 25-OH-vitamin D, and 1,25-(OH)2-vitamin D were measured. RESULTS: Both PCOS and increased body weight had a significant positive effect on serum PTH values. PTH concentrations were significantly correlated with age, BMI, glucose, PRL, SHBG, and testosterone. Only the correlations with testosterone and PRL were BMI independent. The effect of PCOS on PTH concentrations remained significant after adjustment for BMI, but not after adjustment for testosterone concentration. Increased body weight also had a significant negative effect on 25-OH- and 1,25 (OH)2-vitamin D concentrations, but no association with the syndrome was observed. CONCLUSIONS: The results of the present study are in agreement with previous data supporting an association of increased PTH and decreased vitamin D metabolite concentrations with obesity. Moreover, the present findings indicate, for the first time, that PTH probably is also linked to PCOS-associated hyperandrogenism. PMID- 16037413 TI - DNA sequencing by indexer walking. AB - BACKGROUND: There is a need for DNA sequencing methods that are faster, more accurate, and less expensive than existing techniques. Here we present a new method for DNA analysis by means of indexer walking. METHODS: For DNA sequencing by indexer walking, we ligated double-stranded synthetic oligonucleotides (indexers) to DNA fragments that were produced by type IIS restriction endonucleases, which generate nonidentical 4-nucleotide 5' overhangs. The subsequent amplification (30 thermal cycles) of indexed DNA provided a template for automated DNA sequencing with fluorescent dideoxy terminators. The data gathered in the first sequencing reaction permitted further movement into the unknown nucleotide sequence by digestion of analyzed DNA with selected type IIS restriction endonuclease followed by ligation of the next indexer. A library of presynthesized indexers consisting of 256 oligonucleotides was used for bidirectional analysis of DNA molecules and provided universal primers for sequencing. RESULTS: The proposed protocol was successfully applied to sequencing of cryptic plasmids isolated from pathogenic strains of Escherichia coli. The overall error rate for base-calling was 0.5%, with a mean read length of 550 nucleotides. Approximately 1000 nucleotides of high-quality sequence could be obtained per day from a single clone. CONCLUSIONS: Indexer walking can be used as a low-cost procedure for nucleotide sequence determination of DNA molecules, such as natural plasmids, cDNA clones, and longer DNA fragments. It can also serve as an alternative method for gap filling at the final stage of genome sequencing projects. PMID- 16037414 TI - Detection of single-nucleotide polymorphisms by PCR with universal energy transfer-labeled primers: application to folate- and cobalamin-related genes. PMID- 16037415 TI - Pharmacokinetics and metabolism of a new potent antiepileptic drug, 2,2,3,3 tetramethycyclopropanecarbonylurea, in rats. AB - The pharmacokinetics and metabolism of 2,2,3,3 tetramethylcyclopropanecarbonylurea (TMCU), a potent anticonvulsant compound, were studied in male Sprague-Dawley rats following i.v. (5 mg/kg), oral (20 mg/kg), and i.p. (20 mg/kg) administrations. Urine samples were analyzed by gas chromatography-mass spectrometry (GC/MS) and liquid chromatography-mass spectrometry. Plasma samples were analyzed by GC/MS. TMCU absolute bioavailability was 83% and 90% following oral and i.p. dosing, respectively. Following i.p. administration, the peak plasma concentration (C(max)) obtained 45 min after dosing was 15.4 mg/l. Following oral dosing, C(max) was 6.5 mg/l, and it was reached after 4 h. The disposition kinetics of TMCU in rats was adequately described by a one-compartment open body model. TMCU is well distributed into the extravascular tissues with volume of distribution (V(ss)) of 0.87 l/kg and undergoes extensive metabolism. Only a small fraction of TMCU excreted unmetabolized in the urine (6.3 +/- 0.8%). trans-2-Hydroxymethyl-2,3,3 trimethylcyclopropanecarbonylurea (OH-TMCU) was a predominant metabolite of TMCU. Its structure was established by NMR and X-ray crystallography. Following i.p. administration of 5 and 20 mg/kg TMCU, the drug was excreted in the urine as OH TMCU at an extent of 28.3 +/- 2.6% and 42.1 +/- 3.8%, respectively. A portion of OH-TMCU was excreted in the urine as TMCU sulfate and TMCU glucuronide. PMID- 16037416 TI - Multiple causes of high extinction risk in large mammal species. AB - Many large animal species have a high risk of extinction. This is usually thought to result simply from the way that species traits associated with vulnerability, such as low reproductive rates, scale with body size. In a broad-scale analysis of extinction risk in mammals, we find two additional patterns in the size selectivity of extinction risk. First, impacts of both intrinsic and environmental factors increase sharply above a threshold body mass around 3 kilograms. Second, whereas extinction risk in smaller species is driven by environmental factors, in larger species it is driven by a combination of environmental factors and intrinsic traits. Thus, the disadvantages of large size are greater than generally recognized, and future loss of large mammal biodiversity could be far more rapid than expected. PMID- 16037417 TI - Effects of telomerase and telomere length on epidermal stem cell behavior. AB - A key process in organ homeostasis is the mobilization of stem cells out of their niches. We show through analysis of mouse models that telomere length, as well as the catalytic component of telomerase, Tert, are critical determinants in the mobilization of epidermal stem cells. Telomere shortening inhibited mobilization of stem cells out of their niche, impaired hair growth, and resulted in suppression of stem cell proliferative capacity in vitro. In contrast, Tert overexpression in the absence of changes in telomere length promoted stem cell mobilization, hair growth, and stem cell proliferation in vitro. The effects of telomeres and telomerase on stem cell biology anticipate their role in cancer and aging. PMID- 16037418 TI - Control and detection of singlet-triplet mixing in a random nuclear field. AB - We observed mixing between two-electron singlet and triplet states in a double quantum dot, caused by interactions with nuclear spins in the host semiconductor. This mixing was suppressed when we applied a small magnetic field or increased the interdot tunnel coupling and thereby the singlet-triplet splitting. Electron transport involving transitions between triplets and singlets in turn polarized the nuclei, resulting in marked bistabilities. We extract from the fluctuating nuclear field a limitation on the time-averaged spin coherence time T2* of 25 nanoseconds. Control of the electron-nuclear interaction will therefore be crucial for the coherent manipulation of individual electron spins. PMID- 16037419 TI - Mechanisms for the inhibition of DNA methyltransferases by tea catechins and bioflavonoids. AB - In the present investigation, we studied the modulating effects of several tea catechins and bioflavonoids on DNA methylation catalyzed by prokaryotic SssI DNA methyltransferase (DNMT) and human DNMT1. We found that each of the tea polyphenols [catechin, epicatechin, and (-)-epigallocatechin-3-O-gallate (EGCG)] and bioflavonoids (quercetin, fisetin, and myricetin) inhibited SssI DNMT- and DNMT1-mediated DNA methylation in a concentration-dependent manner. The IC(50) values for catechin, epicatechin, and various flavonoids ranged from 1.0 to 8.4 microM, but EGCG was a more potent inhibitor, with IC(50) values ranging from 0.21 to 0.47 microM. When epicatechin was used as a model inhibitor, kinetic analyses showed that this catechol-containing dietary polyphenol inhibited enzymatic DNA methylation in vitro largely by increasing the formation of S adenosyl-L-homocysteine (a potent noncompetitive inhibitor of DNMTs) during the catechol-O-methyltransferase-mediated O-methylation of this dietary catechol. In comparison, the strong inhibitory effect of EGCG on DNMT-mediated DNA methylation was independent of its own methylation and was largely due to its direct inhibition of the DNMTs. This inhibition is strongly enhanced by Mg(2+). Computational modeling studies showed that the gallic acid moiety of EGCG plays a crucial role in its high-affinity, direct inhibitory interaction with the catalytic site of the human DNMT1, and its binding with the enzyme is stabilized by Mg(2+). The modeling data on the precise molecular mode of EGCG's inhibitory interaction with human DNMT1 agrees perfectly with our experimental finding. PMID- 16037420 TI - Health and health care in transitional Europe. PMID- 16037421 TI - The road to reform. PMID- 16037423 TI - Mental health in post-communist countries. PMID- 16037422 TI - Supply and regulation of medicines. PMID- 16037424 TI - Combating non-communicable diseases. PMID- 16037425 TI - Blind eye to complaints allowed psychiatrists to abuse patients. PMID- 16037426 TI - Lack of targets leads to underperformance in NHS services. PMID- 16037428 TI - 25,000 civilians have been killed in Iraq since invasion. PMID- 16037430 TI - Professor Roy Meadow struck off. PMID- 16037439 TI - Swedish court rules against doctor at centre of row over destroyed research data. PMID- 16037440 TI - NHS publicises advances in antenatal and neonatal screening programmes. PMID- 16037441 TI - European court upholds restriction on sale of food supplements. PMID- 16037442 TI - WHO needs more money to deliver better health care to poor people. PMID- 16037443 TI - Lawyers will appeal ruling that cleared Guantanamo doctor of ethics violations. PMID- 16037446 TI - Association of socioeconomic position with insulin resistance among children from Denmark, Estonia, and Portugal: cross sectional study. AB - OBJECTIVES: To examine the association between socioeconomic position and insulin resistance in children from three countries in northern Europe (Denmark), eastern Europe (Estonia), and southern Europe (Portugal) that have different physical, economic, and cultural environments. DESIGN: Cross sectional study. PARTICIPANTS: 3189 randomly selected schoolchildren aged 9 and 15 years from Denmark (n = 933), Estonia (n = 1103), and Portugal (n = 1153). MAIN OUTCOME MEASURE: Insulin resistance (homoeostasis model assessment). RESULTS: Family income and parental education were inversely associated with insulin resistance in Danish children but were positively associated with insulin resistance in Estonian and Portuguese children. Among Danish children, insulin resistance was 24% lower (95% confidence interval -38% to -10%) in those whose fathers had the most education compared with those with the least education. The equivalent results were 15% (2% to 28%) higher for Estonia and 19% (2% to 36%) higher for Portugal. These associations remained after adjustment for a range of covariates: -20% (-36% to -5%) for Denmark, 10% (-4% to 24%) for Estonia, and 18% (-1% to 31%) for Portugal. Strong statistical evidence supported differences between the associations in Denmark and those in the other two countries in both unadjusted and adjusted models (all P < 0.03). CONCLUSIONS: Among Danish children, those with the most educated and highest earning parents had least insulin resistance, whereas the opposite was true for children from Estonia and Portugal. PMID- 16037447 TI - Health inequalities and social dynamics in Europe. PMID- 16037448 TI - Changes in dietary fat and declining coronary heart disease in Poland: population based study. PMID- 16037449 TI - Will Europe's agricultural policy damage progress on cardiovascular disease? PMID- 16037450 TI - Lung cancer mortality at ages 35-54 in the European Union: ecological study of evolving tobacco epidemics. PMID- 16037451 TI - Making the transition to action. PMID- 16037452 TI - Comparison of amount of biomedical research originating from the European Union and the United States. AB - OBJECTIVE: To examine and compare the research productivity of the European Union, the four "candidate" countries (those currently waiting to join the EU), and the United States in several fields of biomedical sciences. DESIGN: A retrospective observational study-bibliometric analysis. DATA SOURCES: Manuscripts published by authors from each country separately and from each group of countries for the period 1994 to 2004 and included in the Essential Science Indicators database of the Institute of Scientific Information. MAIN OUTCOME MEASURES: Number of published articles and number of citations, adjusted for gross domestic product and population size. RESULTS: 1,485,749 articles were published by authors from the EU compared with 1,356,805 from the US. The research productivity of the first 15 countries to join the EU, adjusted for population, was lower (76%) than that of the US-and even lower (66%) when the 10 newest EU countries were included in the analysis. CONCLUSION: The newest EU members and the EU candidate countries need further help and resources to increase their productivity, thereby improving the productivity of the EU as a whole. PMID- 16037453 TI - Bridging the gaps in biomedical research. PMID- 16037454 TI - Survey of attitudes and knowledge about science in medical students in southeast Europe. PMID- 16037455 TI - Factors associated with spousal physical violence in Albania: cross sectional study. AB - OBJECTIVE: To describe the prevalence of intimate partner violence and associated factors among married women in Albania. DESIGN: Cross sectional study. SETTING: Tirana, the capital city of Albania. PARTICIPANTS: A representative sample of 1039 married women aged 25-65 living in Tirana and recorded in the 2001 census. METHODS: Questionnaire on intimate partner violence and social and demographic characteristics of the women and their husbands. MAIN OUTCOME MEASURE: Women's experience within the past year of being hit, slapped, kicked, or otherwise physically hurt by the husband. RESULTS: More than a third (37%, 384/1039) of women had experienced violence. Risk was greatest among women aged 25-34 (odds ratio 1.47, 95% CI 1.04 to 2.09), women with tertiary education (3.70, 2.04 to 6.67), women in white collar jobs (4.0, 1.59 to 10.0), women with least educated husbands (5.01, 2.91 to 8.64), and women married to men raised in rural areas (3.31, 2.29 to 4.80). Women were at higher risk if they were more educated than their husbands (4.76, 2.56 to 9.09). CONCLUSIONS: In transitional Albania, the risk of spousal violence is high, and more empowered women are at greater risk. PMID- 16037456 TI - Development of family medicine in Kosovo. PMID- 16037457 TI - Implementing general practice in Russia: getting beyond the first steps. PMID- 16037458 TI - Cardiovascular risk factors in Croatia: struggling to provide the evidence for developing policy recommendations. PMID- 16037459 TI - Public health reforms in Estonia: impact on the health of the population. PMID- 16037460 TI - Transition and public health in the Slovak Republic. PMID- 16037461 TI - Impact of transition on public health in Ukraine: case study of the HIV/AIDS epidemic. PMID- 16037462 TI - HIV is not just a transitional problem. PMID- 16037463 TI - Transition and the HIV risk environment. PMID- 16037464 TI - Croatian healthcare system in transition, from the perspective of users. PMID- 16037465 TI - Patients' empowerment: the East side story. PMID- 16037466 TI - Ethics and the structures of health care in the European countries in transition: hospital ethics committees in Croatia. PMID- 16037467 TI - Ethics committees and countries in transition: a figleaf for structural violence? PMID- 16037468 TI - Ethics in health care and research in European transition countries: reality and future prospects. PMID- 16037469 TI - Managing healthcare reform in Hungary: challenges and opportunities. PMID- 16037470 TI - A tale of two Germanys: East Germany's health system provides lessons 14 years on. PMID- 16037471 TI - A tale of two Germanys: Some patients in former East Germany were more equal than others. PMID- 16037472 TI - A tale of two Germanys: Health problems in eastern Germany are clearly related to transition. PMID- 16037473 TI - Perindopril monotherapy and PROGRESS in Europe. PMID- 16037474 TI - Poland is losing its doctors. PMID- 16037475 TI - Health care based on priorities is lost in decentralisation. PMID- 16037476 TI - From the former Soviet Union: Socioeconomic health inequalities exist in post communist Lithuania. PMID- 16037477 TI - From the former Soviet Union: Maternal education seems to determine pregnancy outcomes in Russia. PMID- 16037478 TI - TB in immigrants is not public health risk, but uncontrolled epidemics are. PMID- 16037479 TI - Nuclear weapons are another post-communist health hazard. PMID- 16037480 TI - Georgia to Georgia initiative. PMID- 16037482 TI - Hearing, deaf, and hard-of-hearing Israeli adolescents' evaluations of deaf men and deaf women's occupational competence. AB - This study examined 74 deaf and hard-of-hearing (D/HH) and 91 hearing high school students regarding their own occupational aspirations and their evaluations of occupational competence (EOCs) for deaf adults. In the EOC, participants rated the suitability of 25 occupations (varying according to prestige and required level of communication) for deaf men and women. The results showed that occupations requiring intensive communication levels, regardless of their prestige, were evaluated as much less suitable for deaf individuals than were those requiring less communication. D/HH adolescents did not find highly prestigious occupations as suitable for deaf adults even when communication barriers were irrelevant. Both D/HH and hearing participants expressed biased evaluations of deaf women's competence, but no further evidence emerged for stereotypic attitudes. Higher educational aspirations among hearing adolescents, especially hearing males, correlated with a higher EOC of deaf adults. No such associations emerged for D/HH participants. No gender effects emerged. Implications of these outcomes for career development, especially for females, were discussed. PMID- 16037481 TI - Development and pilot testing of a new psychosocial intervention for older Latinos with chronic psychosis. AB - Latinos constitute the largest minority in the United States, and there is an increasing number of Latino patients with schizophrenia and other psychoses living into old age. However, few specific behavioral interventions have been developed aimed at improving the functioning of this group. We evaluated a psychosocial intervention designed to improve the everyday living skills of middle-aged and older outpatients with very chronic psychotic disorders. Three psychiatric clinics, specializing in care of Latinos, were randomly assigned to (1) a 24-session intervention entitled Programa de Entrenamiento para el Desarrollo de Aptitudes para Latinos (PEDAL) group therapy (n = 21) targeting areas identified in our previous work as being problematic for this population (e.g., using public transportation) or (2) a time-equivalent friendly support group (SG; n = 8). Compared to the patients randomized to SG, PEDAL-treated patients' performance on everyday living skills improved significantly postintervention and was still significantly better at a 6-month maintenance follow-up period and at a 12-month no-treatment follow-up period. There was no significant change in psychopathology. Limitations of this pilot study are discussed. Results suggest that participation in this skills training program, designed specifically for older Latino patients with long-standing psychotic disorders, has the potential to significantly increase the patients' independence and improve functional skills. PMID- 16037483 TI - Nonverbatim captioning in Dutch television programs: a text linguistic approach. AB - In the Netherlands, as in most other European countries, closed captions for the deaf summarize texts rather than render them verbatim. Caption editors argue that in this way television viewers have enough time to both read the text and watch the program. They also claim that the meaning of the original message is properly conveyed. However, many deaf people demand verbatim subtitles so that they have full access to all original information. They claim that vital information is withheld from them as a result of the summarizing process. Linguistic research was conducted in order (a) to identify the type of information that is left out of captioned texts and (b) to determine the effects of nonverbatim captioning on the meaning of the text. The differences between spoken and captioned texts were analyzed on the basis of on a model of coherence relations in discourse. One prominent finding is that summarizing affects coherence relations, making them less explicit and altering the implied meaning. PMID- 16037484 TI - Inhibition of tumor necrosis factor-alpha-induced RANTES secretion by alkaline protease in A549 cells. AB - Pseudomonas aeruginosa is a gram-negative bacterium that is an opportunistic pathogen in patients with cystic fibrosis and in immunocompromised hosts. This bacterium produces a variety of proteolytic enzymes, including alkaline protease (AP), which has multiple biological effects. This study investigated the effects of AP on the A549 pulmonary epithelial cell line. Results demonstrate that AP inhibited tumor necrosis factor (TNF)-alpha-induced RANTES gene expression and secretion in a concentration-dependent manner. The TNF-alpha-induced RANTES gene expression and secretion was attenuated with a neutralizing monoclonal antibody directed against the TNF receptor type 1 (TNFR1). Conversely, a neutralizing monoclonal antibody directed against TNF receptor type II had no effect, suggesting that these events were regulated through the TNFR1 receptor. In addition, we observed that soluble TNF receptor type 1 (sTNFR1) levels were significantly increased in culture supernatants of AP-treated cells in a concentration-dependent manner. Finally, membrane-associated TNFR1 was decreased after AP exposures. In these studies, the enzymatically inactive form of AP had no effect on TNF-alpha-induced RANTES secretion, shedding of sTNFR1, or membrane associated TNFR1. These results demonstrate that AP stimulates shedding of cell surface TNFR1, resulting in an increase in sTNFR1. Consequently, these events decrease the cells' ability to stimulate RANTES gene expression and secretion through TNFR1. PMID- 16037485 TI - Expression and role of the hyaluronan receptor RHAMM in inflammation after bleomycin injury. AB - Lung injury is associated with increased concentrations of hyaluronan (hyaluronic acid, HA). HA modifies cell behavior through interaction with cell-associated receptors such as receptor for HA-mediated motility (RHAMM, CD168). Using a function blocking anti-RHAMM antibody (R36), we investigated the expression and role of RHAMM in the inflammatory response to intratracheal bleomycin in rats. Immunostaining showed increased expression of RHAMM in macrophages 4-7 d after injury. Surface biotin labeling of cells isolated by lavage confirmed increased surface expression of a 70-kD RHAMM after lung injury, and in situ hybridization demonstrated increased RHAMM mRNA in macrophages responding to injury. Time-lapse cinemicrography demonstrated a 5-fold increase in motility of alveolar macrophages from bleomycin-treated animals that was completely blocked by R36 in vitro. Further, HA-stimulated macrophage chemotaxis was also inhibited by R36. Daily administration of R36 to injured animals resulted in a 40% decrease in macrophage accumulation 7 d after injury. Further, H&E staining of tissue sections showed that bleomycin-mediated changes in lung architecture were improved with R36 treatment. Taken together with previous results showing the inhibitory effects of HA-binding peptide on inflammation and fibrosis, we conclude that the interaction of RHAMM with HA is a critical component of the recruitment of inflammatory cells to the lung after injury. PMID- 16037486 TI - Engraftment of neonatal lung fibroblasts into the normal and elastase-injured lung. AB - Interstitial fibroblasts are an integral component of the alveolar wall. These cells produce matrix proteins that maintain the extracellular scaffold of alveolar structures. Emphysema is characterized by airspace enlargement resulting from the loss of alveolar cellularity and matrix. In this study, we explored the endotracheal delivery of fibroblasts to the lung parenchyma as a means of repairing damaged alveolar structures directly or indirectly for the delivery of transgenes. Fibroblasts were isolated from the lungs of neonatal transgenic mice expressing GFP during the period of rapid alveolarization. These GFP+ cells maintained their myofibroblast phenotype in culture and expressed elastin and alpha-smooth muscle actin mRNA. We administered GFP+ fibroblasts to saline- and elastase-treated mice by endotracheal instillation. We detected more GFP+ fibroblasts in the alveolar walls and in the interstitial areas of elastase injured lungs than in normal lungs as assessed by immunohistochemistry and fluorescent imaging. The presence of GFP+ fibroblasts in the interstitium demonstrated transepithelial migration of these cells. Expression of GFP+ fibroblasts in recipient lungs was maintained for at least 20 d after endotracheal administration. These cells synthesize matrix components including elastin in vitro and could contribute to restoring the structural integrity of the alveolar wall. PMID- 16037487 TI - Cloning and characterization of the phosphoglucomutase of Trypanosoma cruzi and functional complementation of a Saccharomyces cerevisiae PGM null mutant. AB - Trypanosoma cruzi is the etiological agent of Chagas' disease, a chronic illness characterized by progressive cardiomyopathy and/or denervation of the digestive tract. The parasite surface is covered with glycoconjugates, such as mucin-type glycoproteins and glycoinositolphospholipids (GIPLs), whose glycans are rich in galactopyranose (Galp) and/or galactofuranose (Galf) residues. These molecules have been implicated in attachment of the parasite to and invasion of mammalian cells and in modulation of the host immune responses during infection. In T. cruzi, galactose (Gal) biosynthesis depends on the conversion of uridine diphosphate (UDP)-glucose (UDP-Glc) into UDP-Gal by an NAD-dependent reduction catalyzed by UDP-Gal 4-epimerase. Phosphoglucomutase (PGM) is a key enzyme in this metabolic pathway catalyzing the interconversion of Glc-6-phosphate (Glc-6 P) and Glc-1-P which is then converted into UDP-Glc. We here report the cloning of T. cruzi PGM, encoding T. cruzi PGM, and the heterologous expression of a functional enzyme in Saccharomyces cerevisiae. T. cruzi PGM is a single copy gene encoding a predicted protein sharing 61% amino acid identity with Leishmania major PGM and 43% with the yeast enzyme. The 59-trans-splicing site of PGM RNA was mapped to a region located at 18 base pairs upstream of the start codon. Expression of T. cruzi PGM in a S. cerevisiae null mutant-lacking genes encoding both isoforms of PGM (pgm1Delta/pgm2Delta) rescued the lethal phenotype induced upon cell growth on Gal as sole carbon source. PMID- 16037488 TI - Hypoglycosylation with increased fucosylation and branching of serum transferrin N-glycans in untreated galactosemia. AB - Untreated classic galactosemia (galactose-1-phosphate uridyltransferase [GALT] deficiency) is known as a secondary congenital disorders of glycosylation (CDG) characterized by galactose deficiency of glycoproteins and glycolipids (processing defect or CDG-II). The mechanism of this undergalactosylation has not been established. Here we show that in untreated galactosemia, there is also a partial deficiency of whole glycans of serum transferrin associated with increased fucosylation and branching as seen in genetic glycosylation assembly defects (CDG-I). Thus galactosemia seems to be a secondary "dual" CDG causing a processing as well as an assembly N-glycosylation defect. We also demonstrated that in galactosemia patients, transferrin N-glycan biosynthesis is restored upon dietary treatment. PMID- 16037489 TI - Monomer/dimer equilibrium of the AB-type lectin from mistletoe enables combination of toxin/agglutinin activities in one protein: analysis of native and citraconylated proteins by ultracentrifugation/gel filtration and cell biological consequences of dimer destabilization. AB - The biological activity of a lectin is influenced by its quaternary structure. Viscumin is special among the family members of toxic AB-type plant lectins, because it triggers mitogenicity, toxicity, and agglutination. Its activity profile is dependent on the concentration, motivating a thorough inspection of the status of quaternary structure. Over a broad range of protein concentrations (0.01-25 mg/mL), viscumin occurs as a dimer. At high concentrations, the solutions exhibited nonideality, self-association, and polydispersity in sedimentation equilibrium and velocity experiments caused by irreversible aggregation. Calculation of viscumin's overall shape based on sedimentation velocity data resulted in an elongated dimer form resembling that of crystallized agglutinin. Appearance of monomers was restricted to concentrations in the submicrogram/mL level, as demonstrated by fast protein liquid chromatography gel filtration analysis. To shift the equilibrium to the monomer for comparative cell biological assays, we performed chemical modification under conditions protecting the lectin activity. Citraconylation was effective to destabilize the dimer. Binding studies by fluorescence-activated cell scan analysis revealed a reduction in cell association upon modification and a tendency for increased sensitivity towards haptenic inhibitors at microg/mL concentrations. Nonetheless, growth inhibition continued to be potent for the ricin-like monomer despite reduced extent of binding. Occurrence of a concentration-dependent monomer/dimer equilibrium appears to achieve the same objectives as the development of two separate protein entities in Ricinus communis, an alternative strategy to emergence of a monomeric toxin, and cell cross-linking dimeric agglutinin. PMID- 16037490 TI - Differential analysis of site-specific glycans on plasma and cellular fibronectins: application of a hydrophilic affinity method for glycopeptide enrichment. AB - Isolation of glycopeptides utilizing hydrogen bonding between glycopeptide glycans and a carbohydrate-gel matrix in the organic phase is useful for site specific characterization of oligosaccharides of glycoproteins, when combined with mass spectrometry. In this study, recovery of glycopeptides was improved by including divalent cations or increasing the organic solvent in the binding solution, without losing specificity, whereas it was still less effective for those with a long peptide backbone exceeding 50 amino acid residues. The method was then applied to the analysis of glycan heterogeneities at seven N glycosylation sites in each of the plasma and cellular fibronectins (FNs). There was a remarkable site-specific difference in fucosylation between these isoforms; Asn1244 selectively escaped the global fucosylation of cellular FN, whereas only Asn1007 and Asn2108 of the plasma isoform underwent modification. In addition, a new O-glycosylation site was identified at Thr279 in the connecting segment between the fibrin- and heparin-binding domain and the collagen-binding domain, and the glycopeptide was reactive to a peanut agglutinin lectin. Considering that another mucin-type O-glycosylation site lies within a different connecting segment, the O-glycosylation of FN was suggested to play a significant role in segregating the neighboring domains and thus maintaining the topology of FN and the domain functions. In addition, the method was applied to apolipoprotein B-100 (apoB100) whose N-glycan structures at 17 of 19 potential sites have been reported, and characterized the remaining sites. The results also demonstrated that the enriched glycopeptide provides resources for site-specific analysis of oligosaccharides in glycoproteomics. PMID- 16037491 TI - Patients with unsolved congenital disorders of glycosylation type II can be subdivided in six distinct biochemical groups. AB - Defects in the biosynthesis of N- and core 1 O-glycans may be found by isoelectric focusing (IEF) of plasma transferrin and apolipoprotein C-III (apoC III). We hypothesized that IEF of transferrin and apoC-III in combination with sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of apoC-III may provide a classification for congenital disorders of glycosylation (CDG) patients. We analyzed plasma from 22 patients with eight different and well characterized CDG subtypes and 19 cases with unsolved CDG. Transferrin IEF (TIEF) has been used to distinguish between N-glycan assembly (type 1 profile) and processing (type 2 profile) defects. We differentiated two different CDG type 2 TIEF profiles: The "asialo profile" characterized by elevated levels of asialo- and monosialotransferrin and the "disialo profile" characterized by increased levels of disialo- and trisialotransferrin. ApoC-III IEF gave two abnormal profiles ("apoC-III(0)" and "apoC-III(1)" profiles). The results for the eight established CDG forms exactly matched the theoretical expectations, providing a validation for the study approach. The combination of the three electrophoretic techniques was not additionally informative for the CDG-Ix patients as they had normal apoC-III IEF patterns. However, the CDG-IIx patients could be further subdivided into six biochemical subgroups. The robustness of the methodology was supported by the fact that three patients with similar clinical features ended in the same subgroup and that another patient, classified in the "CDG-IIe subgroup," turned out to have a similar defect. Dividing the CDG-IIx patients in six subgroups narrows down drastically the options of the primary defect in each of the subgroups and will be helpful to define new CDG type II defects. PMID- 16037492 TI - Structures and mechanisms of glycosyltransferases. AB - Glycosyltransferases (GTs) catalyze the transfer of a sugar moiety from an activated donor sugar onto saccharide and nonsaccharide acceptors. A sequence based classification spreads GTs in many families thus reflecting the variety of molecules that can be used as acceptors. In contrast, this enzyme family is characterized by a more conserved three-dimensional architecture. Until recently, only two different folds (GT-A and GT-B) have been identified for solved crystal structures. The recent report of a structure for a bacterial sialyltransferase allows the definition of a new fold family. Progress in the elucidation of the structures and mechanisms of GTs are discussed in this review. To accommodate the growing number of crystal structures, we created the 3D-Glycosyltransferase database to gather structural information concerning this class of enzymes. PMID- 16037493 TI - Agrin binds alpha-synuclein and modulates alpha-synuclein fibrillation. AB - Recent studies have begun to investigate the role of agrin in brain and suggest that agrin's function likely extends beyond that of a synaptogenic protein. Particularly, it has been shown that agrin is associated with the pathological lesions of Alzheimer's disease (AD) and may contribute to the formation of beta amyloid (Abeta) plaques in AD. We have extended the analysis of agrin's function in neurodegenerative diseases to investigate its role in Parkinson's disease (PD). Alpha-synuclein is a critical molecular determinant in familial and sporadic PD, with the formation of alpha-synuclein fibrils being enhanced by sulfated macromolecules. In the studies reported here, we show that agrin binds to alpha-synuclein in a heparan sulfate-dependent (HS-dependent) manner, induces conformational changes in this protein characterized by beta-sheet structure, and enhances insolubility of alpha-synuclein. We also show that agrin accelerates the formation of protofibrils by alpha-synuclein and decreases the half-time of fibril formation. The association of agrin with PD lesions was also explored in PD human brain, and these studies shown that agrin colocalizes with alpha synuclein in neuronal Lewy bodies in the substantia nigra of PD brain. These studies indicate that agrin is capable of accelerating the formation of insoluble protein fibrils in a second common neurodegenerative disease. These findings may indicate shared molecular mechanisms leading to the pathophysiology in these two neurodegenerative disorders. PMID- 16037494 TI - Roles for the galactose-/N-acetylgalactosamine-binding lectin of Entamoeba in parasite virulence and differentiation. AB - Entamoeba histolytica, an intestinal protozoan parasite, is a major cause of morbidity and mortality in developing countries. The pathology of the disease is caused by the colonization of the large intestine by the amoebic trophozoites and the invasion of the intestinal epithelium. Some of the trophozoites will eventually differentiate into the infectious cyst form, allowing them to be transmitted out of the bowel and into water supplies to be passed from person to person. Both the virulence of the organism and the differentiation process relies on a galactose-/N-acetylgalactosamine (GalNAc)-binding lectin that is expressed on the surface of trophozoites. The functional activity of this lectin has been shown to be involved in host cell binding, cytotoxicity, complement resistance, induction of encystation, and generation of the cyst wall. The role of the lectin in both differentiation and virulence suggests that it may be a pivotal molecule that determines the severity of the infection from a commensal state resulting from increased encystation to an invasive state. The lectin-glycan interactions that initiate these diverse processes are discussed with emphasis on comparing the binding of host ligands and the interactions involved in encystation. PMID- 16037496 TI - Multivariate statistical methods. PMID- 16037497 TI - Axial: an abused word? AB - OBJECTIVE: This article will discuss how the word axial has developed into a synonym for transverse, and why this is incorrect usage when describing an anatomic plane. CONCLUSION: Radiologists should be aware of what these terms mean and use them appropriately. Journal editors are in a unique position to encourage this. PMID- 16037498 TI - Comparison of contrast-enhanced MRI with iodine-123 BMIPP for detection of myocardial damage in hypertrophic cardiomyopathy. AB - OBJECTIVE: The purpose of this study was to compare contrast-enhanced MRI with dual-radionuclide SPECT for the detection of myocardial damage associated with hypertrophic cardiomyopathy. SUBJECTS AND METHODS: Twenty-three patients with hypertrophic cardiomyopathy were examined. Delayed hyperenhancement of the damaged myocardium was observed using contrast-enhanced MRI, and regional wall thickness and left ventricular ejection fraction were measured using cine balanced steady-state free precession MRI. Dual-radionuclide SPECT using technetium-99m sestamibi and iodine-123 15-(p-iodophenyl)-3-(R,S) methylpentadecanoic acid (BMIPP) was performed at rest. In the abnormal myocardial segments, agreement between the contrast-enhanced MRI and 123I BMIPP SPECT was assessed. The relationships between the regional and global cardiac abnormalities and the delayed hyperenhancement on MRI and decreased uptake of 123I BMIPP were also evaluated. RESULTS: In 368 left ventricular segments, 57 segments showed delayed hyperenhancement on MRI, 43 segments showed decreased uptake of 123I BMIPP, and seven showed decreased uptake of (99m)Tc sestamibi. The delayed hyperenhancement and decreased uptake of 123I BMIPP were frequently observed in the interventricular septal wall (p < 0.0001); however, the agreement between the methods in detecting myocardial abnormalities was fair (kappa = 0.38). The abnormal septal walls were significantly thicker than those without apparent abnormalities (p = 0.031). There was an inverse correlation between the number of enhancing segments and the ejection fraction (r = -0.53). CONCLUSION: In hypertrophic cardiomyopathy, contrast-enhanced MRI was valuable for the detection of extensive myocardial damage. PMID- 16037499 TI - Two-phase reconstruction for the assessment of left ventricular volume and function using retrospective ECG-gated MDCT: comparison with echocardiography. AB - OBJECTIVE: The aims of our study were to investigate the clinical feasibility of a two-phase reconstruction method based on ECG to evaluate left ventricular (LV) volume and function using cardiac MDCT and to compare these results with those from echocardiography. SUBJECTS AND METHODS: The LV end-diastolic and end systolic volumes, stroke volume, and ejection fraction were measured using two different methods of cardiac MDCT in 19 patients who had undergone cardiac MDCT and echocardiography. The first was a two-phase reconstruction method based on retrospective ECG-triggering: The end-systolic phase was reconstructed when the reconstruction window was located halfway in the ascending T wave on ECG, and the end-diastolic phase was reconstructed when the reconstruction window was located at the starting point of the QRS complex on ECG. The second was a multiphase reconstruction method: 20 series of images were reconstructed at every 5% throughout the cardiac cycle. The LV volumes and function determined by the two reconstruction methods were compared. The results measured by cardiac MDCT were compared with those obtained by echocardiography. RESULTS: The LV end-diastolic and end-systolic volumes, stroke volume, and ejection fraction measured by the two-phase reconstruction method correlated well with those measured by the multiphase reconstruction method (r = 0.984, 0.978, 0.969, 0.969, respectively). There were no significant differences between the results of the two different reconstruction methods (p > 0.05). The LV volumes showed moderate to good correlation between cardiac MDCT and echocardiography (0.766 < r < 0.940). Ejection fraction measured by cardiac MDCT yielded a significant overestimation of 2.9% +/- 8.7% (mean +/- SD) compared with that measured by echocardiography. CONCLUSION: A two-phase reconstruction method on cardiac MDCT is relatively simple and can provide an objective standard for reconstructing the appropriate image sets for end-diastole and end-systole without the need to review serial preview images. PMID- 16037500 TI - Anomalous origin of the left coronary artery from the pulmonary artery in adulthood on CT and MRI. PMID- 16037501 TI - CT angiography of pulmonary artery aneurysms in Hughes-Stovin syndrome. PMID- 16037502 TI - Pulmonary sarcoidosis: comparison of findings of inspiratory and expiratory high resolution CT and pulmonary function tests between smokers and nonsmokers. AB - OBJECTIVE: The purpose of our study was to compare the high-resolution CT and pulmonary function test findings of smokers and nonsmokers with pulmonary sarcoidosis. MATERIALS AND METHODS: Full inspiratory and expiratory high resolution CT of the thorax and pulmonary function tests were performed in 46 patients (23 smokers and 23 lifelong nonsmokers) with histologically proven sarcoidosis. The median interval between high-resolution CT and pulmonary function tests was 8 days (range, 0-27 days). High-resolution CT findings were categorized into six patterns, and the overall extent of each pattern was scored independently (high-resolution CT score). Correlation between each high resolution CT score with each pulmonary functional parameter was performed using Spearman's rank correlation and stepwise multiple regression analysis. RESULTS: Air trapping on expiration (45/46 patients, 98%) and small nodules on inspiration (all 46 patients, 100%) were the most common findings. Smokers had a greater extent of emphysema than nonsmokers (p = 0.002). No significant difference was seen in the extent of air trapping, consolidation, ground-glass attenuation, reticular opacities, or small nodules between smokers and nonsmokers. On Spearman's rank correlation, the extent of air trapping negatively correlated with forced vital capacity in smokers (p < 0.05) but not in nonsmokers. The extent of small nodules negatively correlated with forced vital capacity and positively correlated with the ratio of forced expiratory volume in 1 sec to forced vital capacity in nonsmokers (p < 0.05, both) but not in smokers, respectively. On stepwise multiple regression analysis, the extent of air trapping on CT was independently associated with decreased forced vital capacity (p < 0.05), and cigarette smoking was the main determinant of decrease in maximum midexpiratory flow and forced expiratory flow at 50% of vital capacity (p < 0.01). CONCLUSION: Cigarette smoking confounds the correlation between the CT and pulmonary function test findings in patients with sarcoidosis. Therefore, smoking history must be taken into account when correlating the extent of parenchymal sarcoidosis on CT with functional impairment. PMID- 16037503 TI - Comparison of eight different digital chest radiography systems: variation in detection of simulated chest disease. AB - OBJECTIVE: In a short period, a variety of technically different digital radiography chest systems have become available for clinical use. Our purpose was to assess the diagnostic performance of eight different digital radiography chest systems for detection of simulated chest disease under clinical conditions. MATERIALS AND METHODS: Assessed were four different flat-panel detector systems, two different charge-coupled device systems, one selenium-coated drum, and one storage phosphor system. For each system, 10 chest images of an anthropomorphic chest phantom were obtained. Each image contained one to 12 simulated chest lesions. Eight radiologists performed soft-copy interpretations. Entrance dose was measured and effective dose calculated. A semi-parametric logistic regression model was used for statistical analysis. RESULTS: Statistically significant differences were found in the diagnostic performance of the eight digital chest systems (p = 0.01). Best performance was observed with the charge-coupled device system with slot-scan technology, yielding a sensitivity of 46% (132 of 288) lesions detected. The performance of three flat-panel detectors and the selenium drum system was not significantly different from the slot-scan charge-coupled device system. Fewer lesions were detected with the storage phosphor system than with all other digital technologies, with a sensitivity of 34% (99 of 288) lesions detected, slot-scan charge-coupled device system versus storage phosphor system, p < 0.001. The effective dose varied among the digital systems. CONCLUSION: We found differences in diagnostic performance among the eight different digital chest systems. Differences in detection rates are predominantly explained by detector design. PMID- 16037504 TI - Endobronchial actinomycosis associated with broncholithiasis: CT findings for nine patients. AB - OBJECTIVE: Our objective was to assess the CT findings for endobronchial actinomycosis associated with broncholithiasis. CONCLUSION: Endobronchial actinomycosis associated with broncholithiasis manifests as a proximal obstructive calcified endobronchial nodule associated with distal post obstructive pneumonia of the involved lobe or segment on CT. The possibility of endobronchial actinomycosis should be entertained when broncholithiasis is seen on CT in tuberculosis-endemic areas. PMID- 16037505 TI - Radiologic and pathologic features of bronchiolitis. AB - OBJECTIVE: The purpose of this article is to describe and illustrate the clinical, pathologic, and imaging features of the inflammatory and fibrotic forms of bronchiolitis. The CT features presented in this article represent the typical features associated with each entity. CONCLUSION: Direct signs of bronchiolitis include centrilobular nodules and tree-in-bud pattern. Indirect signs include mosaic attenuation and air trapping. Although classic examples of each entity exist, there can be substantial overlap in the appearances, and distinguishing among these entities is not always possible. When high-resolution CT features overlap, clinical details will usually help to narrow the differential diagnosis. PMID- 16037506 TI - Contracture of the deltoid muscle: sonographic evaluation with MRI correlation. AB - OBJECTIVE: This article evaluates the sonographic features of deltoid contracture (DC) with MRI correlation. MATERIAL AND METHODS: Two reviewers evaluated the imaging features in 22 painful shoulders of 20 patients with a sonographic diagnosis of DC and a subsequent confirming MRI study. The sonographic and MRI findings with regard to the lesion extent (assessed by a 3-point scale: 1 = less than or equal to one third of the longitudinal deltoid length involved, 2 = greater than one third and less than or equal to two thirds involved, and 3 = greater than two thirds involved), transverse lesion morphologic appearance, and maximal transverse diameter measured were compared with kappa statistics and Wilcoxon's signed rank test, respectively. RESULTS: Compared with MRI, there were two false-positive diagnoses of DC on sonography. Among the 20 true-positive diagnoses, sonography showed good agreement with MRI in assessing the lesion extent (kappa = 0.796, p < 0.001). Three sonographic lesion morphologic patterns for hyperechoic lesions (I = with multiple < 8-mm hypoechoic spots, II = heteroechoic lesions with predominant 8-15-mm hypointense areas, and III = > 15 mm calcified nodules, respectively) showed excellent agreement with three MRI lesion patterns (I = multiple < 8-mm hypointense spots, II = predominant 8-15-mm hypointense areas, and III = > 15-mm hypointense nodules, respectively) (kappa = 0.921, p < 0.001). However, the maximum lesion diameters appeared significantly larger on sonography than on MRI (2.8 +/- 0.6 cm vs 2.0 +/- 0.8 cm, mean +/- SD; p < 0.001), which was plausibly ascribed to the better sonographic delineation of hyperechoic immature fibrotic tissues. CONCLUSION: Sonography is helpful for evaluating DC and correlates well with MRI. PMID- 16037507 TI - Comparison of fat-suppressed T2-weighted fast spin-echo sequence and modified STIR sequence in the evaluation of the rotator cuff tendon. AB - OBJECTIVE: This study was performed to determine whether a modified version of the classic STIR sequence provides similar information about the integrity of the rotator cuff tendon as the commonly used fat-suppressed T2-weighted fast spin echo sequence. SUBJECTS AND METHODS: Sixty-one consecutive MRI examinations of the shoulder in 57 patients were performed using a coronal oblique T1-weighted spin-echo sequence, a modified version of the STIR sequence, and a fat-suppressed T2-weighted fast spin-echo sequence. Three reviewers independently assessed the rotator cuff tendon using the coronal oblique modified inversion recovery sequence and T1-weighted spin-echo sequence. After a minimum of 4 weeks, reviewers assessed the rotator cuff tendon using the fat-suppressed T2-weighted fast spin-echo sequence and T1-weighted spin-echo sequence. The kappa statistic was used to measure the degree of concordance between interpretations when each sequence was used independently. The conditional probability that a full- and a partial-thickness tear would be diagnosed on both sequences was calculated. Image quality was assessed in a side-by-side comparison. RESULTS: The overall weighted kappa score was 0.82, which indicates excellent concordance between the two sequences. If a full-thickness tear of the rotator cuff tendon was found on the fat-suppressed T2-weighted fast spin-echo sequence, there was a 94.1% probability that the same conclusion would be reached using the modified inversion recovery sequence. If a partial-thickness tear was found on the fat-suppressed T2-weighted fast spin-echo sequence, there was an 80.3% probability that the same conclusion would be reached with the modified inversion recovery sequence. Fat suppression in the modified inversion recovery sequence was superior to that in the T2 weighted fast spin-echo sequence in 26-39% of the examinations. CONCLUSION: The modified inversion recovery sequence and fat-suppressed T2-weighted fast spin echo sequence provide similar information about the integrity of the rotator cuff tendon. PMID- 16037508 TI - Fat-suppressed 3D spoiled gradient-echo MRI and MDCT arthrography of articular cartilage in patients with hip dysplasia. AB - OBJECTIVE: Our objective was to assess the diagnostic ability of MDCT arthrography for acetabular and femoral cartilage lesions in patients with hip dysplasia. MATERIALS AND METHODS: A disorder of the articular cartilage was evaluated in 20 hips of 18 patients with acetabular dysplasia who did not have osteoarthritis or who had early stage osteoarthritis before undergoing pelvic osteotomy surgery. The findings on fat-suppressed 3D fast spoiled gradient-echo MRI and MDCT arthrography of the hip were evaluated by two independent observers, and sensitivity, specificity, and accuracy were determined using arthroscopic findings as the standard of reference. Kappa values were calculated to quantify the level of interobserver agreement. RESULTS: The sensitivity and specificity for the detection of any cartilage disorder (grade 1 or higher) were (observer 1/observer 2) 49%/67% and 89%/76%, respectively, on MRI, and 67%/67% and 89%/82%, respectively, on CT arthrography. The sensitivity and specificity for the detection of cartilage lesions with substance loss (grade 2 or higher) were (observer 1/observer 2) 47%/53% and 92%/87%, respectively, on MRI, and 70%/79% and 93%/94%, respectively, on CT arthrography. CT arthrography provided significantly higher sensitivity in the detection of grade 2 or higher lesions than MRI for both observers. Interobserver agreement in the detection of grade 2 or higher cartilage lesions was moderate (kappa = 0.53) on MRI and substantial (kappa = 0.78) on CT. CONCLUSION: MDCT arthrography is a sensitive and reproducible method for assessing articular cartilage lesions with substance loss in patients with hip dysplasia. PMID- 16037509 TI - T1-weighted MRI characteristics of pedal osteomyelitis. AB - OBJECTIVE: The objective of our study was to better define the T1-weighted MRI characteristics of surgically proven pedal osteomyelitis. CONCLUSION: Decreased T1 marrow signal in a geographic medullary distribution with a confluent pattern and concordance with fat-suppressed T2- and T1-weighted postcontrast signal abnormality was present in 100% of the surgically proven cases of pedal osteomyelitis. None of the patients with decreased T1 marrow signal in a subcortical distribution or in a hazy, reticulated pattern had surgically proven osteomyelitis regardless of the fat-suppressed T2-weighted or postcontrast T1 weighted findings. PMID- 16037510 TI - Resorption of osteochondroma by accompanying pseudoaneurysm. PMID- 16037511 TI - MRI, arthroscopy, and histologic observations of an annular ligament causing painful snapping of the elbow joint. PMID- 16037512 TI - Needle tract implantation after sonographically guided percutaneous biopsy of hepatocellular carcinoma: evaluation of doubling time, frequency, and features on CT. AB - OBJECTIVE: The purpose of this study was to evaluate the doubling time, frequency, and features on dynamic CT of extrahepatic needle tract implantation of malignant neoplasms after sonographically guided percutaneous biopsy for hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Between January 1997 and June 2003, 1,055 patients underwent sonographically guided percutaneous biopsy for HCC. The serial changes of implanted tumor volume were estimated on retrospective review of CT, and their doubling times were calculated from the two CT scans showing the first and last visible implanted tumors. The frequency of extrahepatic needle tract implantation of malignant neoplasms was evaluated overall and according to the type of needle used. The CT features of the implanted tumors were examined with regard to size, number, location, morphology, and enhancement pattern. RESULTS: The mean doubling time of extrahepatic needle tract implantation of malignant neoplasms after sonographically guided percutaneous biopsy was 112 days (range, 22-415 days). The mean time interval between biopsy and the emergence of the implanted tumor on CT was 267 days (range, 116-619 days). The overall frequency was 0.76% (8/1,055). The frequencies according to the type of needle were 1.3% (8/622) for the group treated with the end-cutting needle and 0% (0/433) for the group treated with the tru-cut needle; these frequencies differ from each other with statistical significance (p < 0.05, Fisher's exact test). Fifteen of the 17 implanted tumors were round or oval enhancing nodules along the needle tract, and 13 showed persistent enhancement on equilibrium phase images. CONCLUSION: The doubling times of extrahepatic needle tract implantation of malignant neoplasms after sonographically guided percutaneous biopsy for HCC were similar to those of typical HCCs in the liver on CT-based analysis. The frequency was relatively low, and their CT features were similar to those reported previously. PMID- 16037513 TI - CT evaluation of appendicitis and its complications: imaging techniques and key diagnostic findings. AB - OBJECTIVE: This article reviews various CT protocols for appendicitis, identifies key CT findings for diagnosing appendicitis, discusses unusual manifestations such as chronic and recurrent appendicitis, and profiles imaging features that differentiate appendicitis from other inflammatory and neoplastic ileocecal conditions. Patients were studied with helical CT. CONCLUSION: CT is a highly accurate, noninvasive test for appendicitis, but the optimal CT technique is controversial. Major complications of appendicitis (perforation, abscess formation, peritonitis, bowel obstruction, septic seeding of mesenteric vessels, gangrenous appendicitis) and their management are discussed. Abdominal CT is a well-established technique in the study of acute abdominal pain and has shown high sensitivity and specificity for diagnosing and differentiating appendicitis, providing an accurate diagnosis in the early stages of disease. PMID- 16037514 TI - Rapid automated measurement of body fat distribution from whole-body MRI. AB - OBJECTIVE: The purpose of this article is to determine the feasibility of using computer-assisted diagnosis (CAD) techniques to automatically identify, localize, and measure body fat tissue from a rapid whole-body MRI examination. CONCLUSION: Whole-body MRI in conjunction with CAD allows a fast, automatic, and accurate approach to body fat measurement and localization and can be a useful alternative to body mass index. Whole-body fat analysis can be achieved in less than 5 min. PMID- 16037515 TI - Preoperative MRI of potential living-donor-related liver transplantation using a single dose of gadobenate dimeglumine. AB - OBJECTIVE: This article evaluates the feasibility of single-dose gadobenate dimeglumine-enhanced MRI as both an angiographic and biliary contrast medium for making a preoperative evaluation of the donor candidates for a living-donor related liver transplantation. SUBJECTS AND METHODS: Eleven right hepatic lobe donors underwent MRI examinations using T1- and T2-weighted imaging and T2 weighted MR cholangiography (MRC). The MR angiography (MRA) and contrast-enhanced (CE) T1-weighted MRC images then were obtained after injecting a single dose of gadobenate dimeglumine. One radiologist and one surgeon prospectively reviewed all the MRI examinations for hepatic vascular and biliary abnormalities and compared them with the surgical findings and intraoperative cholangiograms. In addition, two blinded reviewers evaluated the two sets of MRC (T2-weighted MRC set and T2-weighted MRC plus CE-T1-weighted MRC set) retrospectively and recorded the anatomic types of the hilar biliary branching pattern along with their confidence in the interpretation. RESULTS: Prospective analysis detected the following vascular variants: hepatic arterial variation in two patients, portal venous variation in one, and a significantly large accessory hepatic vein (> 5 mm) in one. Biliary variants also were identified in two patients. All the MRI findings on the vascular and biliary anatomy were corroborated intraoperatively. Retrospective analysis showed that the mean diagnostic confidence in the combined set was significantly higher than that of the T2-weighted MRC alone by both reviewers (p < 0.05). CONCLUSION: Obtaining both MRA and CE-T1-weighted MRC is feasible using a single dose of gadobenate dimeglumine. Therefore, gadobenate dimeglumine-enhanced MRI might play a role as a preoperative imaging technique for the vascular and biliary evaluation of potential living donors. PMID- 16037516 TI - MR cholangiopancreatography diagnosis of juxtapapillary duodenal diverticulum simulating a cystic lesion of the pancreas: usefulness of an oral negative contrast agent. PMID- 16037517 TI - Prognostic value of the standardized uptake value in esophageal cancer. AB - OBJECTIVE: On PET, the level of tissue glycolysis can be quantified by the accumulation of fluorine-18-fluorodeoxyglucose expressed as the standardized uptake value (SUV). The aims of this study were to investigate the relation between SUV and the stage of disease and whether SUV can be used to predict resectability and survival in patients with esophageal cancer. CONCLUSION: SUV can be used to predict resectability; however, SUV is not an independent factor that can be used to assess survival in patients with esophageal cancer. PMID- 16037518 TI - Phlebosclerotic colitis: imaging-pathologic correlation. AB - OBJECTIVE: The objective of this study was to analyze the imaging findings of phlebosclerotic colitis in comparison with histologic findings. CONCLUSION: Calcifications and/or obstructions of the veins of the colonic wall and adjacent mesentery and collateral formation, edematous thickening of the colonic wall, and increased density in the fatty tissue of the surrounding mesentery were features of this rare entity. However, in the early stage of the disease, no definite calcification of the affected veins was observed. PMID- 16037519 TI - Contrast-enhanced MR angiography and digital subtraction angiography in living renal donors: diagnostic agreement, impact on decision making, and costs. AB - OBJECTIVE: The objective of our study was to evaluate the diagnostic agreement, the impact on decision making, and the costs of contrast-enhanced MR angiography and digital subtraction angiography in the workup of living renal donors. CONCLUSION: Contrast-enhanced MR angiography for the preoperative evaluation of renal donors is superior to digital subtraction angiography in revealing vascular anomalies and depicting parenchymal abnormalities and is less costly; furthermore, it does not lead to preoperative decisions that differ from those based on digital subtraction angiography. If contrast-enhanced MR angiography does not provide sufficient information to make a confident decision, an additional digital subtraction angiography examination should be performed. PMID- 16037521 TI - Mycotic aortic aneurysm presenting initially as an aortic intramural air pocket. PMID- 16037520 TI - Prospective comparison of indirect CT venography versus venous sonography in ICU patients. AB - OBJECTIVE: The purpose of our study was to prospectively compare indirect CT venography (CTV) and duplex venous sonography in the evaluation of acute deep venous thrombosis (DVT) in ICU patients with suspected pulmonary embolism. SUBJECTS AND METHODS: All ICU patients undergoing CT pulmonary angiography in the evaluation of acute pulmonary embolism during a 10-month period were included in the study group. Indirect CTV was performed 3 min after initiation of the contrast bolus and was compared with duplex venous sonography of the lower extremities. Both techniques were compared with a clinical standard when DVT was considered present if a positive result was obtained on sonography or on CTV in the setting of pulmonary embolism. RESULTS: The study group comprised 61 ICU patients. Using the clinical standard, DVT was detected in 10 of 61 patients. The sensitivity and specificity for CTV were 70% and 96%, respectively, and for sonography were 70% and 100%. CONCLUSION: The performance characteristics of CTV and deep venous sonography were similar when compared with a clinical standard. The results support the use of indirect CTV after CT pulmonary angiography as an alternative to sonography in the ICU population. PMID- 16037522 TI - CT imaging findings of pulmonary neoplasms after treatment with radiofrequency ablation: results in 32 tumors. AB - OBJECTIVE: The purpose of this study is to describe the CT appearance of thoracic neoplasms after treatment with radiofrequency ablation (RFA). MATERIALS AND METHODS: Thirty-two thoracic neoplasms in 26 patients had pulmonary RFA and imaging follow-up. Fourteen neoplasms were primary lung cancer and 18 were metastases. The mean pretreatment neoplasm size was 3.1 cm (range, 1.0-7.0 cm), and the average number of neoplasms treated per patient was 1.2 (range, 1-3). The mean follow-up was 10.1 months (range, 1-30 months). Imaging findings on CT were evaluated by three radiologists and documented by consensus. RESULTS: The most common finding immediately after treatment was peripheral ground-glass opacity surrounding the treated neoplasm, seen in 27 of 32 tumors (84%). This rapidly resolved in all but one patient by the end of the first month. Cavitation was seen in 10 of 32 tumors (31%) on follow-up CT and was most common in neoplasms in the inner two thirds of the lung and adjacent to a segmental bronchus. Sixty percent of the cavitations decreased in size on follow-up scans. Ten of 32 tumors (31%) that did not develop cavitation developed bubble lucencies on follow-up CT. Pleural thickening was found in 12 of 22 (55%) parenchymal neoplasms, and linear opacifications were seen between the treated lesion and adjacent pleura in 14 of 22 parenchymal tumors (64%). Pleural effusions were seen in four patients (15%). Fourteen of 22 tumors (64%) with follow-up imaging at 1 month enlarged from pretreatment CT scans. At 3 and 6 months after RFA, the majority remained stable in size. CONCLUSION: Peripheral ground-glass opacity, cavitation, bubble lucencies, and pleural changes are common findings on CT after RFA. Many treated neoplasms increase in size from baseline on 1- to 3-month follow-up CT scans and then remain stable thereafter. Enlargement of a treated tumor after 6 months is felt to represent local recurrence. Stability of a treated lesion beyond 6 months does not guarantee continued stability. PMID- 16037523 TI - TIPS with expanded polytetrafluoroethylene-covered stent: results of an Italian multicenter study. AB - OBJECTIVE: Our objective is to describe the results of a multicenter prospective trial on the safety and efficacy of transjugular intrahepatic portosystemic shunts (TIPS) using the Viatorr stent-graft. SUBJECTS AND METHODS: From 2001 to 2003, 114 patients (75 men and 39 women; mean age, 59.3 years) with portal hypertension underwent TIPS with the Viatorr stent-graft. Indications for treatment were variceal bleeding (n = 49, 43.0%), refractory ascites (n = 52, 45.6%), hypertensive gastropathy (n = 10, 8.8%), Budd-Chiari syndrome (n = 1, 0.9%), and hepatorenal syndrome (n = 2, 1.7%). Eight patients (7.0%) had Child Pugh class A cirrhosis; 60 (52.6%), Child-Pugh class B; and 46 (40.4%), Child Pugh class C. Patients were monitored by color Doppler sonography and phlebography. RESULTS: The procedure was successful in 113 (99.1%) of 114 patients; in one patient, creation of the track was not feasible. The mean portosystemic pressure gradient decreased from 21.8 to 8.7 mm Hg. Three minor immediate complications (2.6%) occurred (two cases of self-limiting hemoperitoneum and one extrahepatic portal puncture requiring covered stenting). At a mean follow-up of 11.9 months, the overall mortality rate was 31.0% (35/113), with a 30-day mortality rate of 8.8% (10/113). Mortality was significantly higher in patients in Child-Pugh class C with refractory ascites and with post-procedural encephalopathy. Cumulative primary patency rates were 91.9%, 79.9%, and 75.9% at 6, 12, and 24 months' follow-up, respectively. Restenosis occurred in 15 patients (13.3%) within the stent (n = 8, 53.3%) or at the ends of the portal (n = 1, 6.7%) or hepatic (n = 6, 40%) veins and was solved by percutaneous transluminal angioplasty (n = 11), stenting (n = 3), or parallel TIPS (n = 1). The secondary patency rate was 98.2%. Post-procedural encephalopathy occurred in 27 patients (23.9%). CONCLUSION: The Viatorr stent graft is safe and effective in TIPS creation, with high primary patency rates. Covering the entire track up to the inferior vena cava can increase patency. PMID- 16037524 TI - Nonenhancing breast malignancies on MRI: sonographic and pathologic correlation. AB - OBJECTIVE: The purpose of this study was to describe the sonographic appearance and histopathologic basis of those malignancies of the breast that show non enhancement on MRI. CONCLUSION: Breast malignancies with suspicious features on sonography may not be detected on MRI due to non-enhancement of the lesions. In such cases, we suggest the use of tissue sampling to differentiate between benign and malignant breast lesions. PMID- 16037525 TI - Primary desmoid tumor (extraabdominal fibromatosis) of the breast. PMID- 16037526 TI - Invasive ductal carcinoma with fibrotic focus: mammographic and sonographic findings with histopathologic correlation. AB - OBJECTIVE: We sought to define the imaging findings of invasive ductal carcinoma with fibrotic focus and its associated histopathologic correlation. CONCLUSION: Radiologists should be aware of the imaging characteristics of this newly described entity because of the significant prognostic implications and perhaps to prompt the pathologist to assess for the presence of a fibrotic focus at excision. PMID- 16037527 TI - Incidental colonic focal lesions detected by FDG PET/CT. AB - OBJECTIVE: The aim of this study was to assess the performance of FDG PET/CT for the detection of colonic lesions, especially advanced neoplasms (villous or >10 mm adenomas, carcinomas). Because of 18F FDG accumulation in adenomatous polyps, PET using FDG can detect early premalignant colorectal lesions. MATERIALS AND METHODS: FDG PET/CT studies performed for a 1-year period in 1,716 consecutive patients with various malignant diseases, except colorectal cancer, were retrospectively reviewed. PET images obtained 1 hr after FDG injection and non contrast CT images used for attenuation correction were fused for analysis. Of 45 patients showing intense focal colonic FDG uptake, 20 patients (with 21 foci) underwent a colonoscopic investigation, and, when necessary, polyp resection. The intensity of FDG uptake was quantified using the standardized uptake value (SUV(max)). RESULTS: The FDG colonic foci were associated with 18 colonoscopic abnormalities in 15 patients, with no colonic abnormality detected in five patients (false-positive [FP] results). Histopathologic findings revealed advanced neoplasms in 13 patients (13 villous adenomas and three carcinomas) and two cases of hyperplastic polyps. A difference in the mean SUV(max) was found between FP and true-positive colonic FDG foci but was not statistically significant (p = 0.14). CONCLUSION: Presence of a focal colonic FDG uptake incidental finding on a PET/CT scan justifies a colonoscopy to detect (pre )malignant lesions. The fusion of PET and CT images allows an accurate localization of the lesions. PET/CT is a useful tool to differentiate pathologic from physiologic FDG uptake. PMID- 16037528 TI - Conservative versus surgical treatment for complex neonatal ovarian cysts: outcomes study. AB - OBJECTIVE: Prenatally diagnosed complex ovarian cysts are most often managed surgically in an attempt to save the ovary. Nevertheless, published surgical results disclose that most patients undergo oophorectomy or salpingo oophorectomy. We assessed whether a surgical or conservative approach was more appropriate by comparing the long-term outcome of infants treated by both methods. A hypothesis for the cause of complex cysts is presented. CONCLUSION: Clinical evidence questions the use of surgery for asymptomatic complex ovarian cysts. Histologic analysis suggests gonad maldevelopment as the origin of complex neonatal ovarian cysts. PMID- 16037529 TI - Reducing radiation exposure from survey CT scans. AB - OBJECTIVE: The purpose of this study was to focus attention on the technique factors commonly used in survey CT scans (e.g., scout, topogram, or pilot scans) to measure the radiation exposure from typical survey CT scans, to compare their exposure to that of typical chest radiographs, and to explore methods for radiation exposure reduction. MATERIALS AND METHODS: The default survey CT scans on 21 CT scanners, representing three different vendors and 11 different models, were investigated. Exposure measurements were obtained with an ion chamber at isocenter and adjusted to be consistent with standard chest radiographic exposure measurement methods (single posterior-anterior projection). These entrance exposures were compared with those of typical chest radiographs, for which the mean for average-sized adults is 16 mR (4.1 x 10(-6) C/kg). RESULTS: The entrance exposures of the default survey CT scans ranged from 3.2 to 74.7 mR (0.8 to 19.3 x 10(-6) C/kg), which is equivalent to approximately 0.2 to 4.7 chest radiographs. By changing the default scan parameters from 120 kVp to 80 kVp and the tube position from 0 degrees (tube above table) to 180 degrees (tube below table), the entrance exposure for the survey CT scan was reduced to less than that of one chest radiograph for all CT scanners. CONCLUSION: For institutions at which the interpreting radiologists do not rely heavily on the appearance of the survey CT image, we recommend adjusting the technique parameters (kilovoltage and X-ray tube position) to decrease radiation exposure, especially for vulnerable patient populations such as children and young women. PMID- 16037530 TI - 16-MDCT of the posttraumatic pediatric elbow: optimum parameters and associated radiation dose. AB - OBJECTIVE: Our objective was to investigate a low-radiation-dose protocol for 16 MDCT of the posttraumatic pediatric elbow using z-axis automatic tube-current modulation, based on optimum scanning parameters determined in a porcine fracture model, and to report the radiation dose from this technique in nine children with acute elbow trauma. CONCLUSION: For the posttraumatic pediatric elbow, 16-MDCT using z-axis automatic tube-current modulation was optimal at 100 kVp with a noise index of 20 and a minimum amperage of 25 mA. PMID- 16037531 TI - Camptodactyly-arthropathy-coxa vara-pericarditis syndrome versus juvenile idiopathic arthropathy. AB - OBJECTIVE: The objective of our article was to highlight the important clinical and radiographic features of camptodactyly-arthropathy-coxa vara-pericarditis (CACP) syndrome. In particular, we emphasize those features that allow differentiation of CACP syndrome from juvenile idiopathic arthropathy. CONCLUSION: CACP syndrome should be considered in all patients who present with a noninflammatory arthropathy or with "atypical juvenile idiopathic arthritis," particularly if radiographs reveal an absence of erosions. In the correct clinical setting, large acetabular cysts on pelvic radiographs may be considered pathognomonic of CACP syndrome. PMID- 16037532 TI - New findings in idiopathic arterial calcification of infancy detected by MDCT. PMID- 16037533 TI - Chest pain evaluation in the emergency department: can MDCT provide a comprehensive evaluation? AB - OBJECTIVE: The purpose of our study was to determine whether MDCT can provide a comprehensive assessment of cardiac and noncardiac causes of chest pain in stable emergency department patients. SUBJECTS AND METHODS: Patients with chest pain who presented to the emergency department without definitive findings of acute myocardial infarction based on history, physical examination, and ECG were recruited immediately after the initial clinical assessment. For each patient, the emergency department physician was asked whether a CT scan would normally have been ordered on clinical grounds (e.g., to exclude pulmonary embolism). Each consenting patient underwent enhanced ECG-gated 16-MDCT. Ten cardiac phases were reconstructed. The images were evaluated for cardiac (coronary calcium and stenosis, ejection fraction, and wall motion and perfusion) and significant noncardiac (pulmonary embolism, dissection, pneumonia, and so forth) causes of chest pain. Correlation was made between the presence of significant cardiac and noncardiac findings on CT and the final clinical diagnosis based on history, examination, and any subsequent cardiac workup at the 1-month follow-up by a consensus of three physicians. RESULTS: Sixty-nine patients met all criteria for enrollment in the study, of whom 45 (65%) would not otherwise have undergone CT. Fifty-two patients (75%) had no significant CT findings and a final diagnosis of clinically insignificant chest pain. Thirteen patients (19%) had significant CT findings (cardiac, 10; noncardiac, 3) concordant with the final diagnosis. CT failed to suggest a diagnosis in two patients (3%), both of whom proved to have clinically significant coronary artery stenoses. In two patients (3%), CT overdiagnosed a coronary stenosis. Sensitivity and specificity for the establishment of a cardiac cause of chest pain were 83% and 96%, respectively. Overall sensitivity and specificity for all other cardiac and noncardiac causes were 87% and 96%, respectively. CONCLUSION: ECG-gated MDCT appears to be logistically feasible and shows promise as a comprehensive method for evaluating cardiac and noncardiac chest pain in stable emergency department patients. Further hardware and software improvements will be necessary for adoption of this paradigm in clinical practice. PMID- 16037534 TI - Remnants of fetal circulation: appearance on MDCT in adults. AB - OBJECTIVE: The objective of our study was to describe the appearance of the remnants of the fetal circulation in adults on MDCT. CONCLUSION: The use of MDCT allows frequent visualization of the remnants of the umbilical vein, ductus venosus, foramen ovale, ductus arteriosus, and umbilical arteries. PMID- 16037535 TI - Chagas' disease involving the brain and spinal cord: MRI findings. PMID- 16037536 TI - Possible airflow around needle in lung biopsy. PMID- 16037537 TI - Extraadrenal pheochromocytoma: a rare cause of tachycardia and hypertension during percutaneous biopsy. PMID- 16037538 TI - Bird's nest filter migration to the right atrium. PMID- 16037539 TI - Chance fracture of the upper thoracic spine. PMID- 16037540 TI - Treatment of multiple splenic aneurysms by coil embolization. PMID- 16037541 TI - Pseudotumoral appearance of peliosis hepatis. PMID- 16037542 TI - MR urography depiction of fluid-debris levels: a sign of pyonephrosis. PMID- 16037543 TI - In vitro model of acute esophagitis in the cat. AB - We have shown that IL-1beta and IL-6, possibly originating from the mucosa in response to injury, inhibit neurally mediated contraction of esophageal circular muscle but do not affect ACh-induced contraction, reproducing the effect of experimental esophagitis on esophageal contraction. To examine the interaction of mucosa and circular muscle in inflammation, we examined the effect of HCl on in vitro esophageal mucosa and circular muscle. Circular muscle strips, when directly exposed to HCl, contracted normally. However, when circular muscle strips were exposed to supernatants of mucosa incubated in HCl (2-3 h, pH 5.8), contraction decreased, and the inhibition was partially reversed by an IL-6 antibody. Supernatants from the mucosa of animals with in vivo-induced acute esophagitis (AE) similarly reduced contraction. IL-6 levels were higher in mucosal tissue from AE animals than in control mucosa and in AE mucosa supernatants than in normal mucosa supernatants. IL-6 levels increased significantly in normal mucosa and supernatants in response to HCl, suggesting increased production and release of IL-6 by the mucosa. IL-6 increased H2O2 levels in the circular muscle layer but not in mucosa. Exposure of the mucosa to HCl caused IL-1beta to increase only in the mucosa and not in the supernatant. These data suggest that HCl-induced damage occurs first in the mucosa, leading to the production of IL-1beta and IL-6 but not H2O2. IL-1beta appears to remain in the mucosa. In contrast, IL-6 is produced and released by the mucosa, eventually resulting in the production of H2O2 by the circular muscle, with this affecting circular muscle contraction. PMID- 16037544 TI - Expression and function of 5-HT4 receptors in the mouse enteric nervous system. AB - The aim of the current study was to identify enteric 5-HT(4) splice variants, locate enteric 5-HT(4) receptors, determine the relationship, if any, of the 5 HT(4) receptor to 5-HT(1P) activity, and to ascertain the function of 5-HT(4) receptors in enteric neurophysiology. 5-HT(4a), 5-HT(4b), 5-HT(4e), and 5-HT(4f) isoforms were found in mouse brain and gut. The ratio of 5-HT(4) expression to that of the neural marker, synaptophysin, was higher in gut than in brain but was similar in small and large intestines. Submucosal 5-HT(4) expression was higher than myenteric. Although transcripts encoding 5-HT(4a) and 5-HT(4b) isoforms were more abundant, those encoding 5-HT(4e) and 5-HT(4f) were myenteric plexus specific. In situ hybridization revealed the presence of transcripts encoding 5 HT(4) receptors in subsets of enteric neurons, interstitial cells of Cajal, and smooth muscle cells. IgY antibodies to mouse 5-HT(4) receptors were raised, affinity purified, and characterized. Nerve fibers in the circular muscle and the neuropil in ganglia of both plexuses were highly 5-HT(4) immunoreactive, although only a small subset of neurons contained 5-HT(4) immunoreactivity. No 5-HT(4) immunoreactive nerves were detected in the mucosa. 5-HT and 5-HT(1P) agonists evoked a G protein-mediated long-lasting inward current that was neither mimicked by 5-HT(4) agonists nor blocked by 5-HT(4) antagonists. In contrast, the 5-HT(4) agonists renzapride and tegaserod increased the amplitudes of nicotinic evoked excitatory postsynaptic currents. Enteric neuronal 5-HT(4) receptors thus are presynaptic and probably exert their prokinetic effects by strengthening excitatory neurotransmission. PMID- 16037545 TI - Activation of CFTR by ASBT-mediated bile salt absorption. AB - In cholangiocytes, bile salt (BS) uptake via the apical sodium-dependent bile acid transporter (ASBT) may evoke ductular flow by enhancing cAMP-mediated signaling to the cystic fibrosis transmembrane conductance regulator (CFTR) anion channel. We considered that ASBT-mediated BS uptake in the distal ileum might also modulate intestinal fluid secretion. Taurocholate (TC) induced a biphasic rise in the short circuit current across ileal tissue, reflecting transepithelial electrogenic ion transport. This response was sensitive to bumetanide and largely abrogated in Cftr-null mice, indicating that it predominantly reflects CFTR mediated Cl- secretion. The residual response in Cftr-null mice could be attributed to electrogenic ASBT activity, as it matched the TC-coupled absorptive Na+ flux. TC-evoked Cl- secretion required ASBT-mediated TC uptake, because it was blocked by a selective ASBT inhibitor and was restricted to the distal ileum. Suppression of neurotransmitter or prostaglandin release, blocking of the histamine H1 receptor, or pretreatment with 5-hydroxytryptamine did not abrogate the TC response, suggesting that neurocrine or immune mediators of Cl- secretion are not involved. Responses to TC were retained after carbachol treatment and after permeabilization of the basolateral membrane with nystatin, indicating that BS modulate CFTR channel gating rather than the driving force for Cl- exit. TC induced Cl- secretion was maintained in cGMP-dependent protein kinase II deficient mice and only partially inhibited by the cAMP-dependent protein kinase inhibitor H89, suggesting a mechanism of CFTR activation different from cAMP or cGMP signaling. We conclude that active BS absorption in the ileum triggers CFTR activation and, consequently, local salt and water secretion, which may serve to prevent intestinal obstruction in the postprandial state. PMID- 16037546 TI - Extracellular matrix stimulates reactive oxygen species production and increases pancreatic cancer cell survival through 5-lipoxygenase and NADPH oxidase. AB - The extracellular matrix (ECM) facilitates pancreatic cancer cells survival, which is of central importance for pancreatic adenocarcinoma that is highly fibrotic. Here, we show that reactive oxygen species (ROS) mediate the prosurvival effect of ECM in human pancreatic cancer cells. Fibronectin and laminin stimulated ROS production and NADPH oxidase activation in pancreatic cancer cells. Both pharmacological and molecular approaches show that fibronectin stimulated ROS production through activation of NADPH oxidase and NADPH oxidase independent pathways and that 5-lipoxygenase (5-LO) mediates both these pathways. Analyses of the mechanisms of ROS production by ECM proteins and growth factors indicate that activation of NADPH oxidase (Nox4) is a common mechanism employed both by ECM proteins and growth factors to increase ROS in pancreatic cancer cells. We also found that Nox4 is present in human pancreatic adenocarcinoma tissues and that these tissues display membrane NADPH oxidase activity. ECM proteins and growth factors activate NADPH oxidase through different mechanisms; in contrast to ECM proteins, growth factors activate NADPH oxidase through 5-LO independent mechanisms. Inhibition of 5-LO or NADPH oxidase with pharmacological inhibitors of these enzymes and with Nox4 or 5-LO antisense oligonucleotides markedly stimulated apoptosis in cancer cells cultured on fibronectin. Our results indicate that ROS generation via 5-LO and downstream NADPH oxidase mediates the prosurvival effect of ECM in pancreatic cancer cells. These mechanisms may play an important role in pancreatic cancer resistance to treatments and thus represent novel therapeutic targets. PMID- 16037547 TI - Partial calcanectomy for the treatment of recalcitrant heel ulcerations. AB - Pressure ulcers of the heel are a major and growing health-care problem. Although prevention and aggressive local wound care and pressure reduction remain the gold standard for treatment of most heel ulcers, recalcitrant wounds may require surgical intervention. Limb salvage when dealing with heel ulcers remains a challenge. Nine feet (eight patients) that underwent partial calcanectomy for chronic nonhealing heel ulcers were evaluated retrospectively. Complete healing occurred in seven of nine feet. Patients who were ambulatory before surgery remained ambulatory after healing. PMID- 16037548 TI - Effect of regular exercise on the radiographic progression of foot osteoarthritis. AB - Among the elderly, osteoarthritis often causes chronic pain and disability. Although research has addressed the association between exercise and osteoarthritis, few studies have examined the effect of exercise on the radiographic progression of osteoarthritis. We investigated the relationship between ongoing exercise and radiographic progression of foot osteoarthritis. The first metatarsophalangeal and medial cuneiform-first tarsometatarsal joints were assessed. Joint-specific osteoarthritis radiographic progression scores were determined using four assessments: joint space narrowing, osteophytes, sclerosis, and a composite score. This cohort study included a subset of 221 men and women aged 40 to 91 years participating in a community-based osteoarthritis study. Adjusted risk estimates (95% confidence intervals) summarizing the relationship between ongoing exercise and radiographic progression in the first metatarsophalangeal joint ranged from 0.34 (0.11-0.99) for osteophytes to 0.66 (0.23-1.92) for sclerosis; because only eight individuals experienced medial cuneiform-first tarsometatarsal joint progression, these estimates were less stable, ranging from 2.41 (0.49-11.83) for composite to 4.29 (0.11-166.52) for osteophytes. Overall, our findings do not suggest that regular exercise is a risk factor for foot osteoarthritis progression. Future replication studies are warranted to confirm these findings. PMID- 16037549 TI - Use of laterally wedged custom foot orthoses to reduce pain associated with medial knee osteoarthritis: a preliminary investigation. AB - Osteoarthritis of the knee is a common condition that can cause considerable pain and disability. Various forms of lateral wedging may be effective in the treatment of medial compartment osteoarthritis, but it is not known whether incorporating a lateral wedge into a custom-molded foot orthosis will achieve similar results. Therefore, 30 subjects (21 men and 9 women) aged 29 to 77 years (mean +/- SD, 58.1 +/- 11.6 years) with radiographically confirmed medial compartment knee osteoarthritis were issued custom-molded foot orthoses with a 5 degrees lateral heel wedge. Pain levels were recorded using a 100-mm visual analog pain scale on the date of issue of the orthoses (baseline) and again 3 and 6 weeks later. Mean +/- SD pain levels were significantly reduced at 3 weeks (34 +/- 22 mm) and 6 weeks (23 +/- 22 mm) versus baseline (69 +/- 19 mm) (F2 = 39.57). The degree of pain reduction was greater in patients with less severe osteoarthritis. At 6 weeks, all subjects had achieved at least some reduction in pain, and 28 reported that their orthoses were comfortable. This preliminary study indicates that laterally wedged foot orthoses may be beneficial in the treatment of mild-to-moderate osteoarthritis of the medial compartment of the knee. Further investigations using a larger sample, longer follow-up, and a no treatment control group seem warranted. PMID- 16037550 TI - Efficacy of fifth metatarsal head resection for treatment of chronic diabetic foot ulceration. AB - This study compares the potential benefit of fifth metatarsal head resection versus standard conservative treatment of plantar ulcerations in people with diabetes mellitus. Using a retrospective cohort model, we abstracted data from 40 patients (22 cases and 18 controls) treated for uninfected, nonischemic diabetic foot wounds beneath the fifth metatarsal head. There were no significant differences in sex, age, duration of diabetes mellitus, or degree of glucose control between cases and controls. Patients who underwent a fifth metatarsal head resection healed significantly faster (mean +/- SD, 5.8 +/- 2.9 versus 8.7 +/- 4.3 weeks). Patients were much less likely to reulcerate during the period of evaluation in the surgical group (4.5% versus 27.8%). The results of this study suggest that fifth metatarsal head resection is a potentially effective treatment in patients at high risk of ulceration and reulceration. PMID- 16037551 TI - Plantar pressure and radiographic changes to the forefoot after the Austin bunionectomy. AB - We describe the effects of the Austin bunionectomy on plantar pressure distribution and radiographic measurements in the forefoot in 31 subjects (44 feet) with mild-to-moderate hallux valgus deformity and 36 control subjects (36 feet). Plantar pressure measurements before and 24 months after surgery showed peak pressure beneath the hallux reduced to normal values. Peak pressure measurements beneath the first, second, and third metatarsal heads in hallux valgus feet were relatively unchanged after surgery and remained higher than normal values. The operation produced significant decreases in mean preoperative radiographic measurements of hallux abductus, metatarsus primus varus, and first metatarsal protrusion distance in these patients to below-normal values. PMID- 16037552 TI - Treatment of verruca plantaris with a combination of topical fluorouracil and salicylic acid. AB - A medical record review was conducted to determine the clinical outcome and average time to resolution of verruca plantaris in 20 patients treated with twice daily applications of either 0.5% or 5.0% topical fluorouracil combined with topical 17% and 40% salicylic acid. Seven patients used 0.5% fluorouracil, and 13 used 5.0% fluorouracil. All of the lesions were sharply debrided at regular 1- or 2-week intervals. All 20 patients achieved full clinical resolution in a mean +/- SD of 82.5 +/- 56.6 days. Three patients (15%) had recurrent lesions, which subsequently resolved with repeated treatment. Two patients (10%) developed local dermatitis, which resolved with temporary discontinuation of the medication and the addition of a topical corticosteroid. It was observed that the twice-daily application of topical fluorouracil and salicylic acid is a safe and effective treatment for verruca plantaris. PMID- 16037553 TI - Effect of tube angulation on the measurement of intermetatarsal angles. AB - We systematically investigated the effect of tube angulation on angular distortion of the anteroposterior radiograph of the foot. Three-dimensional data from the metatarsals originating from computed tomographic scans of ten healthy volunteers were projected onto the supporting surface at various tube angulations to simulate radiography. The distortion of the intermetatarsal angles decreased from 1.2 degrees to 3.5 degrees at 20 degrees tube angulation to 0.4 degrees to 2.7 degrees at 0 degrees tube angulation. The relatively small improvement in angular measurement using 0 degrees instead of 15 degrees tube angulation would not outweigh the adverse effects of changing the standard radiographic technique. Physician awareness of this source of error when planning surgical therapy seems more important. PMID- 16037554 TI - The amount of rearfoot motion used during the stance phase of walking. AB - The purpose of this study was to determine the proportion of available passive frontal plane rearfoot motion that is used during the stance phase of walking. Data were collected from 40 healthy, asymptomatic volunteer subjects (20 men and 20 women) aged 23 to 44 years. Passive inversion and eversion motion was measured in a non-weightbearing position by manually moving the calcaneus. Dynamic rearfoot motion was referenced to a vertical calcaneus and tibia and was measured using a three-dimensional electromagnetic motion-analysis system. The results indicated that individuals used 68.1% of their available passive eversion range of motion and 13.2% of their available passive inversion range of motion during walking. The clinical implication of individuals' regularly operating at or near the end point of their available rearfoot eversion range of motion is discussed. PMID- 16037555 TI - Low-molecular-weight heparins: an overview for the podiatric physician. AB - Deep venous thrombosis is a common but underdiagnosed medical condition. The epidemiologic features, economic impact, morbidity, and mortality of venous thromboembolism make it imperative that the podiatric physician be familiar with its pathogenesis as well as its pharmacologic treatment. Medical literature rooted in clinical evidence has demonstrated that low-molecular-weight heparins are safe and effective for the prevention and treatment of venous thromboembolism. The primary purpose of this article is to review the pharmacologic characteristics of low-molecular-weight heparins. Dosing recommendations for low-molecular-weight heparins as they apply to the prevention of deep venous thrombosis are presented. Finally, a dosing criteria chart is presented to assist the podiatric physician in prescribing and evaluating low molecular-weight heparins as a therapeutic class. PMID- 16037556 TI - Longitudinal tear of the tibialis anterior tendon. AB - Diagnosis and treatment of longitudinal tears of the tibialis anterior tendon are not well documented in the surgical literature. Described here is successful primary surgical repair of a longitudinally torn tibialis anterior tendon in a 60 year-old woman. PMID- 16037557 TI - Bone matrix therapy for aneurysmal bone cysts. AB - Aneurysmal bone cysts are unique pathologic entities that cause pain and local osseous destruction. Many surgical treatment modalities have been described. This article reports on the case of a 16-year-old high school athlete with left heel pain due to an aneurysmal bone cyst in the calcaneus. Curettage of the bone cyst was performed, and the void was filled with a commercially available mixture of cancellous bone and demineralized bone matrix. Early return to athletic activity was achieved, with no recurrence noted at 3-year follow-up. PMID- 16037558 TI - Congenital malalignment of the great toenails in a pair of monozygotic twins. AB - Congenital malalignment of the great toenails is the lateral deviation of the long axis of nail growth relative to the distal phalanx. The nails grow slowly, with thickening, curving, and transverse ridging. We describe a pair of 3-year old monozygotic female twins with congenital malalignment of the great toenails complicated by ingrowing and paronychia. Although there are a few cases without any family history, congenital malalignment is believed to be inherited through an autosomal-dominant gene of variable expression. This report provides further evidence of the heritability of the disease. PMID- 16037559 TI - Three-dimensional reconstruction of magnetic resonance images of a displaced flexor hallucis longus tendon in hallux valgus. AB - By using three-dimensional magnetic resonance image reconstruction, lateral displacement of the flexor hallucis longus tendon and sesamoid bones was made clearly visible in a living patient. This finding supports a biomechanical model related to disturbed muscle balance at the first metatarsophalangeal joint, which could play an important role in the pathogenesis of hallux valgus and metatarsus primus varus. PMID- 16037560 TI - A new fixation technique for the Lapidus bunionectomy. AB - Presented here is a preliminary report of 102 patients who underwent first metatarsocuneiform joint arthrodeses performed with external fixation for the correction of hallux valgus. The advantages of using external fixation are the ability to initiate early weightbearing, predictable fusion, and removal of all of the hardware postoperatively. In the 102 patients reported here, the average time to initiation of unassisted full weightbearing was 13.1 days. The average time to fusion was 5.3 weeks, with removal of the external fixator at an average of 5.5 weeks postoperatively. There was no incidence of delayed union or nonunion. There was one case of pin-tract irritation, which resolved with appropriate pin care and a short course of oral antibiotics. External fixation is an effective alternative to traditional internal fixation techniques in metatarsocuneiform joint arthrodesis. PMID- 16037561 TI - Endoscopic gastrocnemius recession for the treatment of gastrocnemius equinus. AB - A prospective analysis was conducted of the amount of correction obtained and number of complications resulting from 31 endoscopic gastrocnemius recessions in 28 patients. The average amount of increase in ankle dorsiflexion was 18 degrees. Few complications were encountered, with mild "soreness" and distal "bruising" being reported by four patients. The only other complaint was "weakness." The endoscopic gastrocnemius recession is a minimally invasive technique with a low rate of complications that offers a comparable amount of correction to that of traditional open procedures in the treatment of gastrocnemius equinus. PMID- 16037562 TI - American Podiatric Medical Association Best Walking City Competition, 2004. AB - In 2004, the American Podiatric Medical Association conducted its third annual "Best Walking City Competition." This study improved on the 2002 and 2003 studies by increasing the number of cities competing for the title of "Best Walking City" and by including a variety of new measures of walking activities to provide a more comprehensive and equitable basis for comparing cities. The top 20 best walking cities in 2004 were identified from among the 200 largest cities across the United States. Lists of top cities were also developed by city population size and geographic region and by three different types of walking activities prevalent in each city. PMID- 16037563 TI - A comparison of KOH and culture results from two mycology laboratories for the diagnosis of onychomycosis during a randomized, multicenter clinical trial: a subset study. PMID- 16037564 TI - The Farnesoid X receptor: a molecular link between bile acid and lipid and glucose metabolism. AB - Bile acids are the end products of cholesterol metabolism. They are synthesized in the liver and secreted via bile into the intestine, where they aid in the absorption of fat-soluble vitamins and dietary fat. Subsequently, bile acids return to the liver to complete their enterohepatic circulation. The Farnesoid X receptor (FXR) is a member of the nuclear receptor superfamily and has emerged as a key player in the control of multiple metabolic pathways. On its activation by bile acids, FXR regulates bile acid synthesis, conjugation, and transport, as well as various aspects of lipid and glucose metabolism. This review summarizes recent advances in deciphering the role of FXR in the context of hepatic lipid and glucose homeostasis and discusses the potential of FXR as a pharmacological target for therapeutic applications. PMID- 16037565 TI - Oxidative stress promotes endothelial cell apoptosis and loss of microvessels in the spontaneously hypertensive rats. AB - OBJECTIVE: Endothelial cell apoptosis caused by oxidative stress may lead to the loss of microvessels (rarefaction) in hypertension. We examine here the effects of antioxidants on cell apoptosis and rarefaction. METHODS AND RESULTS: The juvenile spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats were treated with superoxide scavengers, Tempol or Tiron, during growth. After the treatment, oxidative stress status, endothelial cell apoptosis rate, and microvessel length density in skeletal muscle and mesentery were evaluated in comparison with age-matched controls. Untreated 16-week-old SHR had higher oxidative stress (P<0.01) and cell apoptosis rate (P<0.05) and lower microvessel length density (371+/-17 mm/mm3 [P<0.01]) compared with age-matched WKY rats (435+/-15 mm/mm3). In the SHR, but not in WKY rats, systemically applied antioxidants attenuated oxidative stress and cell apoptosis rate (P<0.05 versus untreated controls) and prevented the loss of microvessels (411+/-15 mm/mm3 for Tempol [P<0.01 versus untreated control] and 399+/-17 mm/mm3 for Tiron [P<0.05]). CONCLUSIONS: Antioxidant treatment with cell-permeable superoxide scavengers inhibits endothelial cell apoptosis and prevents microvessel rarefaction in the SHR during growth. PMID- 16037567 TI - Atherosclerotic plaque progression and vulnerability to rupture: angiogenesis as a source of intraplaque hemorrhage. AB - Observational studies of necrotic core progression identify intraplaque hemorrhage as a critical factor in atherosclerotic plaque growth and destabilization. The rapid accumulation of erythrocyte membranes causes an abrupt change in plaque substrate characterized by increased free cholesterol within the lipid core and excessive macrophage infiltration. Neoangiogenesis is associated closely with plaque progression, and microvascular incompetence is a likely source of intraplaque hemorrhage. Intimal neovascularization is predominantly thought to arise from the adventitia, where there are a plethora of pre-existing vasa vasorum. In lesions that have early necrotic cores, the majority of vessels invading from the adventitia occur at specific sites of medial wall disruption. A breech in the medial wall likely facilitates the rapid in-growth of microvessels from the adventitia, and exposure to an atherosclerotic environment stimulates abnormal vascular development characterized by disorganized branching and immature endothelial tubes with "leaky" imperfect linings. This network of immature blood vessels is a viable source of intraplaque hemorrhage providing erythrocyte-derived phospholipids and free cholesterol. The rapid change in plaque substrate caused by the excessive accumulation of erythrocytes may promote the transition from a stable to an unstable lesion. This review discusses the potential role of intraplaque vasa vasorum in lesion instability as it relates to plaque rupture. PMID- 16037566 TI - The effects of leukocyte-type 12/15-lipoxygenase on Id3-mediated vascular smooth muscle cell growth. AB - OBJECTIVE: 12/15-lipoxygenase (12/15-LO) has been implicated in the pathogenesis of vascular disease. Vascular smooth muscle cell (VSMC) proliferation is a key component of the response to injury in vascular disease. The role of 12/15-LO in regulating VSMC proliferation is poorly understood. Id3 has been shown to regulate growth in various cell types and is expressed in VSMCs within atherosclerotic and restenotic lesions. This study examines the role of Id3 in 12/15-LO-mediated VSMC proliferation. METHODS AND RESULTS: Primary aortic VSMCs from leukocyte-type 12/15-LO transgenic, leukocyte-type 12/15-LO knockout (KO), and control mice were plated in equal densities and assayed for growth, Id3 protein expression, and Id3 transcription. Results demonstrated that 12/15-LO transgenic VSMCs grew faster, whereas 12/15-LO KO VSMCs grew slower relative to control VSMCs. Further, pharmacological and molecular inhibition of 12/15-LO resulted in decreased VSMC growth. Western blots demonstrated increased Id3 protein in 12/15-LO transgenic VSMCs, whereas luciferase promoter reporter assays revealed increased Id3 transcription. In addition, overexpression of 12/15-LO increased growth in control cells but not in Id3 KO cells. 12/15-LO transgenic VSMCs demonstrated increased protein kinase C (PKC) activity. Consistent with these data, PKC inhibition decreased Id3 promoter activation. CONCLUSIONS: 12/15 LO is an important mediator of VSMC growth. The growth-promoting effects of 12/15 LO are at least partially mediated through induction of Id3 transcription. PMID- 16037568 TI - Combination of tumor necrosis factor-alpha ablation and matrix metalloproteinase inhibition prevents heart failure after pressure overload in tissue inhibitor of metalloproteinase-3 knock-out mice. AB - Cytokine and extracellular matrix (ECM) homeostasis are distinct systems that are each dysregulated in heart failure. Here we show that tissue inhibitor of metalloproteinase (TIMP)-3 is a critical regulator of both systems in a mouse model of left ventricular (LV) dilation and dysfunction. Timp-3(-/-) mice develop precipitous LV dilation and dysfunction reminiscent of dilated cardiomyopathy (DCM), culminating in early onset of heart failure by 6 weeks, compared with wild type aortic-banding (AB). Timp-3 deficiency resulted in increased TNFalpha converting enzyme (TACE) activity within 6 hours after AB leading to enhanced tumor necrosis factor-alpha (TNFalpha) processing. In addition, TNFalpha production increased in timp-3(-/-)-AB myocardium. A significant elevation in gelatinase and collagenase activities was observed 1 week after AB, with localized ECM degradation in timp-3(-/-)-AB myocardium. Timp-3(-/-)/tnfalpha(-/-) mice were generated and subjected to AB for comparative analyses with timp-3(-/-) AB mice. This revealed the critical role of TNFalpha in the early phase of LV remodeling, de novo expression of Matrix metalloproteinases (MMP)-8 in the absence of TNFalpha, and highlighted the importance of interstitial collagenases (MMP-2, MMP-13, and MT1-MMP) for cardiac ECM degradation. Ablation of TNFalpha, or limiting MMP activity with a synthetic MMP inhibitor (PD166793), each partially attenuated LV dilation and cardiac dysfunction in timp-3(-/-)-AB mice. Notably, combining TNFalpha ablation with MMP inhibition completely rescued heart disease in timp-3(-/-)-AB mice. This study provides a basis for anti-TNFalpha and MMP inhibitor combination therapy in heart disease. PMID- 16037569 TI - Distinct pathways regulate expression of cardiac electrical and mechanical junction proteins in response to stretch. AB - To define mechanisms regulating expression of cell-cell junction proteins, we have developed an in vitro system in which neonatal rat ventricular myocytes were subjected to pulsatile stretch. Previously, we showed that expression of the gap junction protein, connexin (Cx) 43, is increased by approximately 2-fold after 1 hour of stretch, and this response is mediated by stretch-induced secretion of vascular endothelial growth factor (VEGF). Here, we report that the mechanical junction proteins plakoglobin, desmoplakin, and N-cadherin are also upregulated by pulsatile stretch but by a mechanism independent of VEGF or other secreted chemical signals. Stretch-induced upregulation of mechanical junction proteins was blocked by anti-beta1 and anti-beta3 integrin antibodies. Transfection of cells with adenovirus expressing GFP-FRNK, a dominant-negative inhibitor of focal adhesion kinase (FAK)-dependent signaling, blocked stretch-induced upregulation of Cx43 and mechanical junction proteins but did not block the ability of exogenous VEGF to upregulate Cx43 expression. Conditioned medium removed from uninfected cells after stretch increased Cx43 expression when added to nonstretched cells, and this effect was blocked by anti-VEGF antibodies, but stretch-conditioned medium from GFP-FRNK cells had no effect on Cx43 expression. The src kinase inhibitor 4-amino-5-(4-chloro-phenyl)-7-(t-butyl)pyrazolol[3,4 d]pyrimidine blocked stretch-induced upregulation of mechanical junction proteins but not Cx43. Thus, stretch upregulates expression of both electrical and mechanical junction proteins via integrin-dependent activation of FAK. Stretch induced upregulation of Cx43 expression is mediated by FAK-dependent secretion of VEGF. In contrast, stretch-induced upregulation of adhesion junction proteins involves intracellular mechanotransduction pathways initiated via integrin signaling and acting downstream of src kinase. PMID- 16037570 TI - Interleukin-10 suppresses tissue factor expression in lipopolysaccharide stimulated macrophages via inhibition of Egr-1 and a serum response element/MEK ERK1/2 pathway. AB - Atherosclerosis is considered to be an inflammatory disease. Tissue factor (TF), a prothrombotic molecule expressed by various cell types within atherosclerotic plaques, is thought to play an essential role in thrombus formation after atherosclerotic plaque rupture. Recent studies suggest that the antiinflammatory cytokine interleukin-10 (IL-10) has many antiatherosclerotic properties. Therefore, the effects of IL-10 on TF expression in response to inflammation were investigated. Mouse macrophages were stimulated with lipopolysaccharide (LPS) in the presence or absence of IL-10. Pretreatment with IL-10 resulted in a 50% decrease in TF mRNA expression and TF promoter activity. Binding of early growth response gene-1 (Egr-1) to the consensus DNA sequence, a key transcriptional activator of TF expression in response to inflammation, and the expression of Egr 1 mRNA were also inhibited by IL-10. This inhibition was independent of the induction of suppressor of cytokine signaling protein-3 by IL-10. Macrophages that had been transfected with luciferase reporter constructs containing the murine Egr-1 5'-flanking sequence exhibited reduced reporter gene activity in response to LPS stimulation with IL-10 pretreatment. Studies with deletion constructs of the Egr-1 promoter identified the proximal serum response element SRE3 as a potential regulatory site for the IL-10 mediated suppression of Egr-1 expression. Furthermore, activation of the upstream signal-transduction elements, such as mitogen-activated protein kinase kinase (MEK) 1/2, extracellular signal regulated kinase 1/2, and Elk-1 were also inhibited by IL-10 pretreatment. Taken together, these results demonstrate a pathway for the IL-10 mediated inhibition of TF expression during inflammation and may explain the antiatherosclerotic effects of IL-10. PMID- 16037571 TI - Alk3/Bmpr1a receptor is required for development of the atrioventricular canal into valves and annulus fibrosus. AB - Endocardial cushions are precursors of mature atrioventricular (AV) valves. Their formation is induced by signaling molecules originating from the AV myocardium, including bone morphogenetic proteins (BMPs). Here, we hypothesized that BMP signaling plays an important role in the AV myocardium during the maturation of AV valves from the cushions. To test our hypothesis, we used a unique Cre/lox system to target the deletion of a floxed Alk3 allele, the type IA receptor for BMPs, to cardiac myocytes of the AV canal (AVC). Lineage analysis indicated that cardiac myocytes of the AVC contributed to the tricuspid mural and posterior leaflets, the mitral septal leaflet, and the atrial border of the annulus fibrosus. When Alk3 was deleted in these cells, defects were seen in the same leaflets, ie, the tricuspid mural leaflet and mitral septal leaflet were longer, the tricuspid posterior leaflet was displaced and adherent to the ventricular wall, and the annulus fibrosus was disrupted resulting in ventricular preexcitation. The defects seen in mice with AVC-targeted deletion of Alk3 provide strong support for a role of Alk3 in human congenital heart diseases, such as Ebstein's anomaly. In conclusion, our mouse model demonstrated critical roles for Alk3 signaling in the AV myocardium during the development of AV valves and the annulus fibrosus. PMID- 16037572 TI - Evidence in favor of a calcium-sensing receptor in arterial endothelial cells: studies with calindol and Calhex 231. AB - Small increases in extracellular Ca2+ dilate isolated blood vessels. In the present study, the possibility that a vascular, extracellular Ca2+-sensing receptor (CaSR) could mediate these vasodilator actions was investigated. Novel ligands that interact with the CaSR were used in microelectrode recordings from rat isolated mesenteric and porcine coronary arteries. The major findings were that (1) raising extracellular Ca2+ or adding calindol, a CaSR agonist, produced concentration-dependent hyperpolarizations of vascular myocytes, actions attenuated by Calhex 231, a negative allosteric modulator of CaSR. (2) Calindol induced hyperpolarizations were inhibited by the intermediate conductance, Ca2+ sensitive K+ (IKCa) channel inhibitors, TRAM-34, and TRAM-39. (3) The effects of calindol were not observed in the absence of endothelium. (4) CaSR mRNA and protein were present in rat mesenteric arteries and in porcine coronary artery endothelial cells. (5) CaSR and IKCa proteins were restricted to caveolin-poor membrane fractions. We conclude that activation of vascular endothelial CaSRs opens endothelial cell IKCa channels with subsequent myocyte hyperpolarization. The endothelial cell CaSR may have a physiological role in the control of arterial blood pressure. PMID- 16037573 TI - Protein kinase G transmits the cardioprotective signal from cytosol to mitochondria. AB - Ischemic and pharmacological preconditioning can be triggered by an intracellular signaling pathway in which Gi-coupled surface receptors activate a cascade including phosphatidylinositol 3-kinase, endothelial nitric oxide synthase, guanylyl cyclase, and protein kinase G (PKG). Activated PKG opens mitochondrial KATP channels (mitoKATP) which increase production of reactive oxygen species. Steps between PKG and mitoKATP opening are unknown. We describe effects of adding purified PKG and cGMP on K+ transport in isolated mitochondria. Light scattering and respiration measurements indicate PKG induces opening of mitoKATP similar to KATP channel openers like diazoxide and cromakalim in heart, liver, and brain mitochondria. This effect was blocked by mitoKATP inhibitors 5-hydroxydecanoate, tetraphenylphosphonium, and glibenclamide, PKG-selective inhibitor KT5823, and protein kinase C (PKC) inhibitors chelerythrine, Ro318220, and PKC-epsilon peptide antagonist epsilonV(1-2). MitoKATP are opened by the PKC activator 12 phorbol 13-myristate acetate. We conclude PKG is the terminal cytosolic component of the trigger pathway; it transmits the cardioprotective signal from cytosol to inner mitochondrial membrane by a pathway that includes PKC-epsilon. PMID- 16037574 TI - Rapid endothelial cell-selective loading of connexin 40 antibody blocks endothelium-derived hyperpolarizing factor dilation in rat small mesenteric arteries. AB - In resistance arteries, spread of hyperpolarization from the endothelium to the adjacent smooth muscle is suggested to be a crucial component of dilation resulting from endothelium-derived hyperpolarizing factor (EDHF). To probe the role of endothelial gap junctions in EDHF-mediated dilation, we developed a method, which was originally used to load membrane impermeant molecules into cells in culture, to load connexin (Cx)-specific inhibitory molecules rapidly (approximately 15 minutes) into endothelial cells within isolated, pressurized mesenteric arteries of the rat. Validation was achieved by luminally loading cell impermeant fluorescent dyes selectively into virtually all the arterial endothelial cells, without affecting either tissue morphology or function. The endothelial monolayer served as an effective barrier, preventing macromolecules from entering the underlying smooth muscle cells. Using this technique, endothelial cell loading either with antibodies to the intracellular carboxyl terminal region of Cx40 (residues 340 to 358) or mimetic peptide for the cytoplasmic loop (Cx40; residues 130 to 140) each markedly depressed EDHF mediated dilation. In contrast, multiple antibodies directed against different intracellular regions of Cx37 and Cx43, and mimetic peptide for the intracellular loop region of Cx37, were each without effect. Furthermore, simultaneous intra- and extraluminal incubation of pressurized arteries with inhibitory peptides targeted against extracellular regions of endothelial cell Cxs (43Gap 26, 40Gap 27, and (37,43)Gap 27; 300 micromol/L each) for 2 hours also failed to modify the EDHF response. High-resolution immunohistochemistry localized Cx40 to the end of endothelial cell projections at myoendothelial gap junctions. These data directly demonstrate a critical role for Cx40 in EDHF-mediated dilation of rat mesenteric arteries. PMID- 16037575 TI - Vascular remodeling versus vasoconstriction in chronic hypoxic pulmonary hypertension: a time for reappraisal? PMID- 16037576 TI - Beware of cells bearing gifts: cell replacement therapy and arrhythmic risk. PMID- 16037577 TI - Bone morphogenetic proteins in vascular calcification. AB - Vascular calcification is a common problem among the elderly and those with chronic kidney disease (CKD) and diabetes. The process of tunica media vascular calcification in CKD appears to involve a phenotypic change in the vascular smooth muscle cell (VSMC) resulting in cell-mediated mineralization of the extracellular matrix. The bone morphogenetic proteins (BMPs) are important regulators in orthotopic bone formation, and their localization at sites of vascular calcification raises the question of their role. In this review, we will discuss the actions of the BMPs in vascular calcification. Although the role of BMP-2 in vascular calcification is not proven, it has been the most studied member of the BMP family in this disease process. The role of BMP-2 may be through inducing osteoblastic differentiation of VSMCs through induction of MSX 2, or by inducing apoptosis of VSMCs, a process thought critical in the initiation of vascular calcification. Additionally, BMP-2 may be related to loss of regulation of the matrix Gla protein. A second BMP, BMP-7, less studied than BMP-2 may have opposing actions in vascular calcification. In postnatal life, BMP 7 is expressed primarily in the kidney, and expression is diminished by renal injury. BMP-7 is an important regulator of skeletal remodeling and the VSMC phenotype. BMP-7 restores skeletal anabolic balance in animal models of CKD with disordered skeletal modeling, also reducing serum phosphate in the process. BMP-7 also reverses vascular calcification in CKD, and reduction in vascular calcification is due, in part, to reduced serum phosphate, an important inducer of VSMC-mediated vascular mineralization and in part to direct actions on the VSMC. PMID- 16037607 TI - Maternal and neonatal scalp hair concentrations of zinc, copper, cadmium, and lead: relationship to some lifestyle factors. AB - Postpartum scalp hair samples from 82 term-pregnancy mother/ neonate pairs were analyzed for their concentrations of zinc (Zn), copper (Cu), cadmium (Cd), and lead (Pb), using inductively coupled plasma-mass spectrometry. Maternal and neonatal Zn concentrations had geometric means (and 99% confidence intervals) of 122.5 microg/g (117.9--131.5 microg/g) and 146.9 microg (141.5--156.7 microg/g) respectively. Corresponding Cu values were 18.4 microg/g (17.6--23.8 microg/g) and 6.7 microg/g (6.3--7.6 microg/g). Those of Cd were 0.49 microg/g (0.47--0.69 microg/g) in the mothers and 0.57 microg/g (0.55--0.86 microg/g) in the neonates. For Pb, they were 7.95 microg/g (7.60--9.32 microg/g) and 4.56 microg/g (4.39- 5.56 microg/g). Cigarette smoking, despite its relatively low prevalence (19.5%), was associated with lower Zn and higher Cd and Pb concentrations and in lower Zn/Cd and Zn/Pb molar concentration ratios. Smoking also altered interelemental relationships, particularly those of Zn with Cd and Pb and those between Cd and Pb. Smoking frequency appeared to show negative dose-response effects on maternal and neonatal Zn concentrations, Zn/Pb molar concentration ratios, and birth weight. Mothers with a history of oral contraceptive (OC) usage had significantly higher Cu concentrations and lower Zn/Cu molar concentration ratios than non users, with the highest Cu concentrations and lowest Zn/Cu values being associated with third-generation OCs. No similar effects were elicited in the respective neonatal Cu concentrations. Neither alcohol consumption nor prenatal supplementation with iron and/or folic acid had discernible effects on the maternal or neonatal elemental concentrations. The data from this study suggest that in a given population of term-pregnancy mothers and neonates, significant interindividual variations in hair trace element concentrations can occur, irrespective of commonality of general environment, and that lifestyle factors, including cigarette smoking and OC usage history, can be significant contributory factors to such variations. The data are discussed in relation to the effects of smoking-associated exposure to Cd and Pb exposure on Zn availability for placental transfer, as well as on the quantitative maternal Zn supply levels to the fetus resulting from the known tendency of smokers to have lower dietary intakes of Zn. PMID- 16037608 TI - Antioxidant enzyme activities of human peripheral blood mononuclear cells exposed to trace elements. AB - The trace elements copper, zinc, and selenium are important immune modulators and essential cofactors of the antioxidant enzymes. In the present study, the proliferative effect of human peripheral mononuclear cells (PBMCs) that have been exposed to copper, zinc, and selenium and the corresponding activities of antioxidant enzymes, namely superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase, were determined. Zinc and copper stimulated the PBMC proliferation in a dose-dependent manner within the dose range 25-200 micromol/L. SOD and GPx activities in PBMCs exposed to zinc were inhibited, whereas catalase activity was unaffected. All the three antioxidant enzymes in the cells exposed to copper were inhibited. Selenium exerted more potent inhibition of the cell proliferation while causing stimulation of the antioxidant enzymes at the lowest dose (25 micromol/L) than at the highest dose (200 micromol/L) tested. A significant negative correlation was observed between proliferation and antioxidant enzyme (SOD and GPx) activities in trace-element-exposed PBMC. The present findings substantiate the importance of trace elements as immune modulators and the involvement of enzymatic antioxidant system in the immune cell regulation. PMID- 16037609 TI - Monitoring metals in the population living in the vicinity of a hazardous waste incinerator: concentrations in autopsy tissues. AB - This study is a part of a monitoring program for the determination of metals in various human tissues of the population living in the vicinity of a new hazardous waste incinerator (HWI) in Constanti (Tarragona County, Spain). Concentrations of arsenic (As), beryllium (Be), cadmium (Cd), chromium (Cr), mercury (Hg), manganese (Mn), nickel (Ni), lead (Pb), tin (Sn), thallium (Tl), and vanadium (V) were determined in brain, bone, kidney, liver, and lung autopsy samples collected in 2003 from 22 individuals who had been living for at least 10 yr in the area under evaluation. Results were compared with the metal levels obtained in a baseline study, which was performed during the construction of the HWI (1996- 1998). In the present survey, As, Be, Tl, and V levels were not detected in any of the analyzed tissues, while Cr concentrations were very close to the limit of detection. The highest levels of Cd and Hg were found in kidney (17.46 microg/g and 0.23 microg/g, respectively), those of Mn in liver (1.07 microg/g), and those of Ni, Pb, and Sn in bone (1.16 microg/g, 2.11 microg/g, and 0.34 microg/g, respectively). In comparison to the results of the baseline study, a general reduction of most metal concentrations was observed in the current survey. PMID- 16037610 TI - Mitochondrial membrane potential is reduced in copper-deficient C2C12 cells in the absence of apoptosis. AB - Mitochondrial membrane potential is reduced in copper-deficient rat hearts, but it is uncertain if this will lead to the onset of apoptosis. To determine if copper deficiency per se leads to apoptosis, C2C12 cells were made copper deficient by treatment with the copper chelator tetraethylenepentamine (TEPA). In TEPA-treated cells, the activity of Cu, Zn-superoxide dismutase and cytochrome-c oxidase decreased dramatically. The protein levels of nuclear-encoded subunits of the cytochromie-c oxidase decreased, but the mitochondrial-encoded subunits remained unchanged. Decreased mitochondrial membrane potential was indicated in TEPA-treated cells, but further investigation of the potential induction of apoptosis by measuring caspase-3 activity, protein concentrations of Bcl-2 and Bax, and DNA fragmentation suggested that apoptosis is not induced in TEPA treated C2C12 cells. Cells with decreased mitochondrial membrane potential were not destined to apoptosis as a result of copper deficiency. PMID- 16037611 TI - Prevention by Ce3+ of DNA destruction caused by Hg2+ in fish intestine. AB - The interactions between Hg2+, Ce3+, and the mixture of Ce3+ and Hg2+, and DNA from fish intestine in vitro were investigated by using absorption spectrum and fluorescence emission spectrum. The ultraviolet absorption spectra indicated that the addition of Hg2+, Ce3+, and the mixture of Ce3+ and Hg2+ to DNA generated an obviously hypochromic effect. Meanwhile, the peak of DNA at 205.2 nm blue-shifted and at 258.2 nm red-shifted. The size of the hypochromic effect and the peak shift of DNA by metal ion treatments was Hg2+>Hg2++Ce3+>Ce3+. The fluorescence emission spectra showed that with the addition of Hg2+, Ce3+, and the mixture of Ce3+ and Hg2+ the emission peak at about 416.2 nm of DNA did not obviously change, but the intensity reduced gradually and the sequence was Hg2+>Hg2++ Ce3+>Ce3+. Hg2+, Ce3+, and the mixture of Ce3+ and Hg2+ had 1.12, 0.19, and 0.41 binding sites to DNA, respectively; the fluorescence quenching of DNA caused by the metal ions all attributed to static quenching. The binding constants (KA) of binding sites were 8.98 x 104 L/mol and 1.02 x 104 L/mol, 5.12 x 104 L/mol and 1.10 x 103 L/mol, 6.66 x 104 L/mol and 2.36 x 103 L/mol, respectively. The results showed that Ce3+ could relieve the destruction of Hg2+ on the DNA structure. PMID- 16037613 TI - The environment & health: an emerging area of research in India. PMID- 16037612 TI - Purification and characterization of mouse hepatic enzyme that converts selenomethionine to methylselenol by its alpha,gamma-elimination. AB - The objective of this study was to purify and characterize a mouse hepatic enzyme that directly generates CH3SeH from seleno-l-methionine (l-SeMet) by the alpha,gamma-elimination reaction. The l-SeMet alpha,gamma-elimination enzyme was ubiquitous in tissues from ICR mice and the activity was relatively high in the large intestine, brain, and muscle, as well as the liver. Aging and sex of the mice did not have any significant influence on the activity in the liver. The enzyme was purified from the mouse liver by ammonium sulfate precipitation and four kinds of column chromatography. These procedures yielded a homogeneous enzyme, which was purified approx 1000-fold relative to mouse liver extract. Overall recovery was approx 8%. The purified enzyme had a molecular mass of approx 160 kDa with four identical subunits. The Km value of the enzyme for the catalysis of l-SeMet was 15.5 mM, and the Vmax was 0.29 units/mg protein. Pyridoxal 5'-phosphate (pyridoxal-P) was required as a cofactor because the holoenzyme could be resolved to the apoenzyme by incubation with hydroxylamine and reconstituted by addition of pyridoxal-P. The enzyme showed the optimum activity at around pH 8.0 and the highest activity at 50 degrees C; it catalyzed the alpha,gamma-elimination reactions of several analogs such as d,l-homocysteine and l-homoserine in addition to l-SeMet. This enzyme also catalyzed the alpha,beta-elimination reaction of Se-methylseleno-l-cysteine. However, l methionine was inert. Therefore, the purified enzyme was different from the bacterial l-methionine gamma-lyase that metabolizes l-SeMet to CH3SeH, in terms of the substrate specificity. These results were the first identification of a mammalian enzyme that specifically catalyzes the alpha,gamma-elimination reaction of l-SeMet and immediately converts it to CH3SeH, an important metabolite of Se. PMID- 16037614 TI - Paragonimiasis: an emerging public health problem in India. PMID- 16037615 TI - Wheelchair propulsion: a straining form of ambulation. PMID- 16037616 TI - Measurement of psychiatric disability. PMID- 16037617 TI - Insect cell culture in research: Indian scenario. AB - Insect cell cultures are widely used in viral diagnosis and biotechnology, for the production of recombinant proteins, viral pesticides and vaccines as well as in basic research in genetics, molecular biology, biochemistry, endocrinology and virology. Following KRP Singh's pioneering research in 1967, a large number of cell lines from diptera, hemiptera, and lepidopteran insects were established and characterized in India. With the availability of the modern tools in molecular biology and the advancements made in biotechnology, the indigenous cell lines may prove useful in creating a future without biohazardous chemical pesticides as well as producing life saving pharmaceuticals and vaccines for many diseases. This review summarizes information gathered regarding the insect cell lines established so far in India. It also covers the familiarization of the well characterized continuous cell lines and their potential applications. Special attention is given to virus susceptibility of the cell lines, the yield of virus with a comparative analysis with other conventional systems. The potential applications of dipteran and lepidopteran cell lines in agriculture and biotechnology are also briefly discussed for prospective studies. PMID- 16037618 TI - Development of enzyme-linked immunosorbent assay for serodiagnosis of human paragonimiasis. AB - BACKGROUND & OBJECTIVE: Although human paragonimiasis is known to occur in Manipur and Arunachal Pradesh, there is no indigenous immunodiagnostic test available in India. Sputum examination for detection of eggs is less sensitive method for diagnosis of paragonimiasis and unfortunately, the eggs are not always present in the sputum of infected individuals. Due to overlapping clinical manifestations and similarities between X-ray picture in pulmonary paragonimiasis and pulmonary tuberculosis, chances of diagnostic confusion can increase. Therefore, the objective of this study was to develop an ELISA test indigenously for diagnosis of paragonimiasis. METHODS: Somatic (S) and excretory-secretory (ES) antigens of adult trematodes belonging to genus Paragonimus were prepared. Serum samples were collected from 22 confirmed patients of paragonimiasis and from five groups of negative controls. The cut-off points for both types of antigens were calculated using receiver operating characteristic (ROC) curve analysis. The sensitivity, specificity, predictive values and efficiency of the ELISA tests were also calculated. RESULTS: IgG-ELISA test using either S or ES antigens were 100 per cent sensitive yet the utility of S antigen for diagnosis of paragonimiasis seemed to be less reliable because of low specificity (91.3%). On the other hand, ES antigen was more reliable with 100 per cent specificity as no false positive cases were recorded. INTERPRETATION & CONCLUSION: ES antigen can be effectively used for screening large populations for paragonimiasis. In areas where both paragonimiasis and tuberculosis are endemic, suspected subjects should be screened for both the diseases because of chance of mistaken diagnosis. Correct diagnosis will avoid unnecessary costly treatment. PMID- 16037619 TI - Effect of chronic use of different propulsion systems in wheelchair design on the aerobic capacity of Indian users. AB - BACKGROUND & OBJECTIVE: The use of wheelchairs (WC) of ergonomically different propulsion mechanisms may influence the cardiorespiratory capacity of the WC user. The purpose of the present investigation was to observe the impact of chronic use of four ergonomically different propulsion systems and age of the WC users on their aerobic capacity. METHODS: The male subjects (n=77), exclusively using hand rim (n=20), arm crank using both arms (n=22), arm crank using one arm (n=17), and arm lever using one arm (n=18) propelled WCs and 20 able-bodied (AB) subjects as the control group participated They performed maximal exercise test in continuous, step-wise incremental workload at a crank rate of 50 rpm on an arm crank ergometer. The VO(2) and heart rate obtained during 2.45 to 3 min of each test exercise, and VO(2max) and HR(max) were derived respectively. Two-way ANOVA and multiple comparison tests were performed to compare the groups with respect to VO(2max) and HR(max). Age was used as a classificatory variable. RESULTS: The maximal physiological response of the AB subjects was superior to the WC users but the WC user groups did not show any significant differences amongst them. The aerobic capacity decreased with advancement of age and for WC users this is more pronounced. Equations for age predicted VO(2max) and HR(max) were also derived. INTERPRETATION & CONCLUSION: As the WC users used the four distinct propulsion systems, it was anticipated that it would induce variation in the physiological variables of the users. But the results of the study revealed that there was no significant difference. Suggesting that the WC users might have developed certain self-regulatory mechanisms in order to overcome the variation induced by the different propulsion systems. PMID- 16037620 TI - Disability assessment in mental illnesses using Indian Disability Evaluation Assessment Scale (IDEAS). AB - BACKGROUND & OBJECTIVE: Psychiatric disorders cause disability in individuals and pose significant burden on their families. In most of the cases residual disability and poor quality of life continue even after disability evaluation in patients with chronic mental illness in very important. The present study was undertaken to assess and compare the disability in patients with schizophrenia and obsessive-compulsive disorder (OCD) using Indian Disability Evaluation Assessment Scale (IDEAS). METHODS: Patients diagnosed to have schizophrenia and OCD with mild severity of illness were included in the study. Indian Disability Evaluation Assessment Scale (IDEAS) was applied. Disability was assessed in these patients on all domains of IDEAS. RESULTS: Majority of the patients with schizophrenia were from rural areas whereas most of the patients with OCD were from urban background. There was comparable disability in the patients with schizophrenia with duration of illness in the range of 2-5 yr and >5 yr. Significant disability in work and global score was seen in patients of obsessive compulsive disorder with duration of illness >5 yr. Patients with schizophrenia had significantly higher disability in all domains than patients with OCD. INTERPRETATION & CONCLUSION: Schizophrenia causes greater disability than obsessive-compulsive disorder in patients. These illnesses affect all areas of daily functioning leading to greater disability, and thus increasing the burden on the family, pose greater challenge for the rehabilitation of patients and their inclusion in the mainstream of the family and society. Further studies on a larger sample need to be done to confirm the finding. PMID- 16037621 TI - Effect of erythromycin on homocysteine-induced extracellular matrix metalloproteinase-2 production in cultured rat vascular smooth muscle cells. AB - BACKGROUND & OBJECTIVE: Several lines of evidence have shown an association between Chlamydia infection and atherosclerosis, but clinical trials of preventive antibiotic (erythromycin) treatment in patients with coronary artery disease have shown conflicting results. Hyperhomocysteinaemia is an independent risk factor of coronary artery disease and causes an intense remodelling of the extracellular matrix in arterial walls, particularly an elastolysis involving metalloproteinases. In the present study we investigated the effects of erythromycin on the production of homocysteineinduced extracellular matrix metalloproteinase-2 (MMP-2) in cultured rat vascular smooth muscle cells (VSMCs). METHODS: Effects of different concentration of homocysteine (Hcy) (0-5000 micromol/l) on MMP-2 production, and the effects of different erythromycin concentrations (0-10 mmol/l) on homocysteine-induced MMP-2 production in cultured rat VSMCs were studied using gelatin zymography and Western blotting. The changes of MMP-2 under various treatments for 1, 3 and 5 days were also compared. RESULTS: Homocysteine (50-1000 mu mol/l) increased the production of MMP-2 significantly in a dose-dependent manner and reduced the production of MMP-2 at a high level (5000 mu mol/l). Increased production of MMP-2 induced by homocysteine was reduced by extracellularly added erythromycin in a dose-dependent manner. INTERPRETATION & CONCLUSION: Homocysteine increased the production of MMP-2 significantly in a dose-dependent manner. Extracellularly added erythromycin decreased homocysteine-induced MMP-2 secretion. The findings of the present study suggested that the beneficial effect of erythromycin on vascular disease processes might be due to its inhibitory effect on the Hcyinduced production of MMP-2 in VSMCs. PMID- 16037622 TI - A survey of bancroftian filariasis for microfilariae & circulating antigenaemia in two villages of Madhya Pradesh. AB - BACKGROUND & OBJECTIVE: The estimation of filariasis prevalence in Panna district of Madhya Pradesh was so far relied upon clinical and night blood smear examination. However, night blood smear examination fails to detect the infection in individuals having low parasitaemia and cryptic filarial infection. The present study was undertaken to re-evaluate the prevalence of filariasis in two villages of Panna district by Og4C3 ELISA. METHODS: The study was carried out during 2002-2003 in two villages (namely Pista and Taroni) of Panna district, Madhya Pradesh. Clinical examination was performed according to WHO criteria to classify filarial disease. Night blood smears collected during 20:00 to 00:00 h were examined to detect microfilariae (Mf). For estimation of circulating filarial antigen (CFA) by Og4C3 ELISA, 2 ml blood was collected from each individual by venepuncture. RESULTS: With Og4C3 ELISA, 38 per cent serum samples of village Pista (n=332) and 47.7 per cent from village Taroni (n=88) were found positive for CFA. The overall disease rate was (243/ 420) 57.9 per cent by CFA while it was (182/420) 43.3 per cent by night blood smear examination. A total of 14.5 per cent individuals were having cryptic filarial infection detected by CFA. INTERPRETATION & CONCLUSION: The study emphasizes the use of CFA estimation being a sensitive and specific diagnostic tool for the evaluation of the true prevalence of the disease. The high CFA prevalence in the study area necessitates early intervention measures to check its transmission. PMID- 16037623 TI - Influenza activity among the paediatric age group in Chennai. AB - BACKGROUND & OBJECTIVE: Respiratory viral infections have a major impact on public health. Acute respiratory infections largely caused by viruses, are the most common illnesses experienced by otherwise healthy adults and children. Among the respiratory viruses, influenza viruses are known to cause outbreaks globally. Information on the activity of influenza virus in our country is limited and none from Chennai. The present study was carried out to isolate and identify the influenza virus serotypes causing acute respiratory infection in children attending a tertiary care centre at Chennai. METHODS: During January to December 2002, 240 children with acute respiratory infection attending the out patient clinic of Institute of Child Health were included by convenient sampling. Throat swabs were collected from 4 to 5 cases every week. Isolation of influenza virus was attempted by inoculating the sample in Madin Darby Canine Kidney (MDCK) cell line. The isolates were typed by haemagglutination inhibition test and confirmed by immunoflourescence assay. RESULTS: Virus isolation was positive in 30 (12.5%) of the 240 samples. Influenza A/H3N2/Panama/ 2000/99 was the predominant serotype isolated accounting for 24 (80%) of the 30 isolates. Influenza B/Sichuan/379/99 was isolated in 4 (13.33%) and a combination of Influenza A/H3N2 and B/Sichuan in 2 (6.6%) of the isolates. INTERPRETATION & CONCLUSION: Isolation of influenza A and B viruses indicated a significant activity of these viruses in Chennai. Peak activity was observed during and after the first spell of rain. The predominance of A/H3N2/ Panama is an indication that the Indian scenario is similar to the global picture of influenza activity. PMID- 16037624 TI - Comparison of two methods to detect carbapenemase & metallo-beta- lactamase production in clinical isolates. AB - BACKGROUND & OBJECTIVE: Bacterial resistance has greatly hampered effective treatment of patients in clinical settings. Non-fermenting Gram-negative bacilli (NFGNB) are common nosocomial pathogens. In this study we attempted to develop a convenient test for early detection of carbapenemase and metallo-beta-lactamase (MbetaL) production in NFGNB. Lack of sufficient reports from India in this area indicated the need for this study. METHODS: A total of 50 imipenem resistant NFGNB were speciated, and their resistance reconfirmed by disk diffusion and minimum inhibitory concentration (MIC) determination by agar dilution. Two different methods namely modified Hodge and EDTA disk synergy tests were evaluated for carbapenemase and metallo-beta-lactamase (MbetaL) production. RESULTS: Of the 50 imipenem resistant NFGNB, 48 and two respectively fell in the resistant and intermediate range in MIC using agar dilution. Majority of these were Pseudomonas aeruginosa (n=28), followed by Burkholderia cepacia (n=9). The modified Hodge test could detect 28 strains as carbapenemase and MbetaL producers, while the EDTA disk synergy test was able to detect an additional 8 strains producing MbetaL and carbapenemase. INTERPRETATION & CONCLUSION: Pseudomonas aeruginosa was found to be the predominant NFGNB in our hospital setting and EDTA disk synergy could detect more carbapenemase and metallo-beta- lactamase producers compared to modified Hodge test. PMID- 16037625 TI - Bacteriology of orthopaedic wound infections in an Indian Tertiary Care Hospital. PMID- 16037627 TI - Soluble angiopoietin receptor Tie-2 in patients with acute myocardial infarction and its effects on angiogenesis. AB - Tie-2 receptor has been shown to play a role in the neovascularization of tumors, but little is known about the role it may play in acute myocardial infarction (AMI). The aims of this research are (1) To study the variety of soluble Tie-2 (sTie-2) in patients with AMI. (2) To study the effects of recombinant soluble Tie-2/Fc on HUVECs viability and tube formation in vitro. Serum levels of sTie-2 in 27 patients with AMI were measured on admission (day 1), day 2, day 3 and day 7 after onset of chest pain and 28 healthy controls by ELISA. In addition, the viability of HUVECs and tube formation area were measured after stimulated with recombinant Tie-2/Fc chimera. Median level of sTie-2 increased significantly in the AMI patients when compared with the controls and the maximum level appeared at day 2 after onset of AMI. Tie-2/Fc induced EC apoptosis and inhibited HUVEC tube formation in vitro. Results of this study showed that the level of sTie-2 increased in AMI. The effects of Tie-2/Fc on EC viability and tube formation indicated that angiogenesis might be inhibited in the acute phase of AMI. PMID- 16037628 TI - Deep venous thrombosis: behaviour of the polymorphonuclear leukocyte integrin pattern at baseline and after in vitro activation. AB - In a group of 18 subjects with acute deep venous thrombosis (DVT), evidenced by clinical examination and echo-color-Doppler, we examined the phenotypical expression of the polymorphonuclear leukocyte (PMN) beta2-integrins (CD11a, CD11b, CD11c, CD18), obtained by using a flow cytofluorimeter. The evaluation was performed before and after in vitro activation (prolonged for 5 and 15 minutes) with 4-phorbol 12-myristate 13-acetate (PMA) and N-formyl-methionyl-leucyl phenylalanine (fMLP). In DVT subjects, at baseline, the phenotypical expression of CD11b was decreased and that of CD11c was increased when compared with normal controls; no difference was found in CD11a and CD18 expression. In normal subjects PMN activation with both activators led to a constant increase of all PMN adhesion molecules; in DVT subjects CD11b, CD11c and CD18 increased, while CD11a expression did not show any change. These data indicate the presence of a functional alteration in circulating PMN cells from patients with DVT. PMID- 16037629 TI - The role of reactive free radicals in ischemic preconditioning--clinical and evolutionary implications. AB - Ischemic preconditioning is a condition of reduced sensitivity to ischemic damage. This protective state can be induced by exposure to periods of brief, sublethal, ischemia prior to a protracted ischemic event, but, more interestingly, also by administration of specific drugs. Recent studies have emphasized the central role of free radicals (including superoxide anion and nitric oxide) in this process. In line with these observations, studies have demonstrated that also drugs such as organic nitrates, which are able to release nitric oxide (but also the highly oxidant superoxide anion) can induce preconditioning. Starting from our observations made in human experimental models regarding the effects of chronic therapy with organic nitrates, we criticize the hypothesis whereby nitrates might be used to induce a state similar to preconditioning upon chronic exposure. As well, we propose a theory for the evolutionary meaning of ischemic preconditioning based on the hypothesis that, while protective over short periods of time, continuous exposure to oxidant free radicals might be associated with a loss of this protective effect and, in certain cases, with an increased oxidative damage. PMID- 16037630 TI - Microvascular hemodynamic responses to arteriovenous shunting in rat limb. AB - Autologus veins have been used clinically as a bypass conduit for reconstruction of small arteries, but there are few data available for microvascular response to arteriovenous (AV) shunting. This study was aimed to evaluate microvascular hemodynamic changes induced by creating AV anastomosis in rat hindlimb. Using intravital fluorescence videomicroscopy, we measured velocities of red blood cells (RBCs) flowing in the microvascular network in the control state, in the occlusion state where the superficial femoral artery (SFA) was occluded, and in the AV shunting state where the AV anastomosis was opened after occlusion of SFA. RBC velocities were measured in 155 capillaries of 6 rats using a dual window method and a frame-by-frame technique. The mean velocity and the coefficient of variation were 0.61 mm/sec and 0.90 in the control state, 0.34 mm/sec and 1.30 in the occlusion state, 0.83 mm/sec and 1.24 in the AV shunting state, respectively. These indicated that hemodynamic heterogeneity among capillaries increased with decrease in mean velocity following the arterial occlusion, while the AV shunting augmented the heterogeneity with increase in mean velocity. Capillaries with low perfusion (<0.1 mm/sec) or high perfusion (>1.0 mm/sec) were 5.8% or 20.6%, 29.6 or 5.2%, and 22.6 or 30.3% out of all measured capillaries in the control, occlusion and AV shunting conditions, respectively. In conclusion, AV shunting increased capillary perfusion and also its spatial heterogeneity, preferentially inducing high velocity in the microvasculature. PMID- 16037631 TI - Plasma total homocysteine concentrations in obese and non-obese female patients with type 2 diabetes mellitus; its relations with plasma oxidative stress and nitric oxide levels. AB - Hyperhomocysteinemia has been identified as independent risk factor for early atherosclerotic vascular disease. The purpose of our study was to investigate the plasma homocystein (Hcy) concentrations and its relationship with lipid peroxidation as thiobarbituric acid reactive substances (TBARS) and nitric oxide (NOx; nitrite plus nitrate) concentrations in age-matched non-obese (n=55) and obese (n=60) female subjects with type 2 diabetes mellitus. Non-obese diabetic patients have significantly higher plasma tHcy and TBARS (p<0.001 and p<0.001), and significantly lower NOx concentrations than the controls (n=25) (p<0.001). The plasma tHcy and TBARS concentrations were higher and nitric oxide concentrations were lower in obese diabetics than in non-obese diabetics (for each comparison; p<0.001). Correlation analysis demonstrated that there was a significant positive correlation between tHcy and TBARS (r=0.452, p<0.01) in diabetics groups. There was no significant correlation between tHcy and plasma NOx, insulin and blood pressure. We thought that Hcy might have a permissive role on the endothelium damage through free radical generating systems and the presence of obesity the free radical induced-damage has been elevated in diabetic patients. PMID- 16037632 TI - Reduced erythrocyte deformability and hypercoagulability in idiopathic sudden sensorineural hearing loss. AB - OBJECTIVE: Sudden sensorineural hearing loss is a frequent disease whose aetiology is still unknown in about 80% of patients. Aim of this study was to evaluate if haemorheological changes and some indexes of hypercoagulability and hypofibrinolysis are associated with idiopathic sudden sensorineural hearing loss (ISSHL). METHODS: We studied 63 patients with ISSHL and 67 healthy control subjects, matched for age, sex and traditional cardiovascular risk factors. Haemorheological studies were performed by assessing whole blood viscosity (WBV) at 0.512 s(-1) and 94.5 s(-1), plasma viscosity (PLV) and erythrocyte deformability index (DI). To assess whole blood coagulation Sonoclot analysis was performed. Sonoclot variables studied were Sonoclot activated clotting time (SonACT), clot rate and time to peak. Fibrinogen, PAI-1 antigen (ag) and factor VIII:C plasma levels were also measured. RESULTS: WBV, PLV, SonACT, clot rate, time to peak, PAI-1ag and factor VIII:C were significantly altered in patients in comparison with controls (p<0.05). A multivariate analysis (adjusted for traditional cardiovascular risk factors, hematocrit, fibrinogen, haemostatic and haemorheological variables) indicated that WBV at 94.5 s(-1), DI, SonACT, clot rate, PAI-1ag plasma levels and factor VIII:C activity were independently associated with ISSHL (p<0.05). CONCLUSIONS: The observed changes in viscosity, blood clotting and fibrinolysis may contribute, at least in part, to the pathophysiological mechanism of ISSHL. PMID- 16037633 TI - Myocardial oxygen tension during fatal right heart failure following perfluorocarbon and radiographic contrast agent interaction. PMID- 16037634 TI - Vascularization and perfusion of hepatocellular carcinoma: assessment with contrast-enhanced ultrasound using perflutren protein-type A microspheres. AB - PURPOSE: To assess the vascularization and the perfusion within hepatocellular carcinoma (HCC) including treatment-related changes with contrast-enhanced (CE) ultrasound (US). MATERIALS AND METHODS: Twenty-six biopsy-verified HCC lesions (size between 2.5-8 cm, median 3.8 cm) in 20 patients were examined with unenhanced and CE vascular US techniques immediately before selective angiography for transarterial chemoembolization (TACE) as well as immediately after TACE using all of the following modalities: color-coded Doppler sonography (CCDS), power Doppler imaging (PDI), CE pulse inversion harmonic imaging with PDI (PIHI+PDI), and CE coded harmonic angiography (CHA). In CE US studies, perflutren protein-type A microspheres were administered as contrast agent in a single 0.5 ml i.v. bolus diluted in 20 ml 0.9% NaCl. Selective arteriograms and CE computed tomographies were taken for reference purposes. The Wilcoxon test was used for statistical analysis. RESULTS: Intratumoral vessels could be visualized before TACE in 11/26 lesions (42%) with CCDS; in 15/26 (58%) with PDI; in 23/26 (88%) with CE CHA; in 26/26 (100%) with CE PIHI+PDI. Following TACE, the sensitivities were calculated as follows: CCDS 33%; PDI 55%; CE CHA 77%; and CE PIHI+PDI 100%. The corresponding negative predictive values were 74% for CCDS; 81% for PDI; 89% for CE CHA and 100% for CE PIHI+PDI. During the capillary phase, contrast enhancement could be observed in the CHA mode only. CONCLUSION: CE US by means of PIHI+PDI and CHA enables reliable visualization of residual tumor following TACE equivalent to that which is attained with angiography and Contrast Harmonic Imaging with Power Doppler, if perflutren microspheres are used as contrast agent in a single low-dose bolus. PMID- 16037635 TI - Red blood cell deformability in iron deficiency anaemia. AB - In patients with iron deficiency anaemia (IDA) it has been suggested that the shortened erythrocyte lifespan may be in part due to decreased erythrocyte deformability. In order to know whether erythrocyte deformability is decreased in IDA patients, we have determined the erythrocyte Elongation Index (EI) by means of ektacytometric techniques (Rheodyn SSD, Myrenne Gmbh, Germany), in 50 IDA patients and 100 well age and sex matched healthy controls. At the three shear stresses tested, 12, 30 and 60 Pa, IDA patients show statistically lower EI than controls (37.4+/-6.7 vs 48.6+/-2.9; 45.0+/-6.0 vs 54.5+/-2.8; 48.7+/-5.8 vs 57.0+/-2.9 mPa.s, respectively; p<0.001). A statistically significant correlation was found between EI at 12, 30, and 60 Pa and the hematimetric indices (MCV, MCH and MCHC), suggesting that the alteration in surface/volume ratio (shape) which characterizes this kind of microcytic hypocromic anaemia, accounts in part for the decreased EI. Rheodyn SSD, as an ektacytometric technique, is very sensitive to alterations in red blood cell geometry, for what seems to be a useful tool for detecting diminished erythrocyte deformability in IDA patients. PMID- 16037636 TI - Targeted therapies; who detects the target? PMID- 16037637 TI - NMD microarray analysis for rapid genome-wide screen of mutated genes in cancer. AB - Gene mutations play a critical role in cancer development and progression, and their identification offers possibilities for accurate diagnostics and therapeutic targeting. Finding genes undergoing mutations is challenging and slow, even in the post-genomic era. A new approach was recently developed by Noensie and Dietz to prioritize and focus the search, making use of nonsense mediated mRNA decay (NMD) inhibition and microarray analysis (NMD microarrays) in the identification of transcripts containing nonsense mutations. We combined NMD microarrays with array-based CGH (comparative genomic hybridization) in order to identify inactivation of tumor suppressor genes in cancer. Such a "mutatomics" screening of prostate cancer cell lines led to the identification of inactivating mutations in the EPHB2 gene. Up to 8% of metastatic uncultured prostate cancers also showed mutations of this gene whose loss of function may confer loss of tissue architecture. NMD microarray analysis could turn out to be a powerful research method to identify novel mutated genes in cancer cell lines, providing targets that could then be further investigated for their clinical relevance and therapeutic potential. PMID- 16037638 TI - Antigen gene transfer to human plasmacytoid dendritic cells using recombinant adenovirus and vaccinia virus vectors. AB - Recombinant adenoviruses (RAd) and recombinant vaccinia viruses (RVV) expressing tumour-associated antigens (TAA) are used as anti-tumour vaccines. It is important that these vaccines deliver the TAA to dendritic cells (DC) for the induction of a strong immune response. Infection of myeloid DC (MDC) with RAd alone is relatively inefficient but CD40 retargeting significantly increases transduction efficiency and DC maturation. Infection with RVV is efficient without DC maturation. Plasmacytoid dendritic cells (PDC) play a role in the innate immune response to viral infections through the secretion of IFNalpha but may also play a role in specific T-cell induction. The aim of our study was to investigate whether PDC are better targets for RAd and RVV based vaccines. RAd alone hardly infected PDC (2%) while CD40 retargeting did not improve transduction efficiency, but it did increase PDC maturation (25% CD83 positive cells). Accordingly, specific CTL activation by RAd infected PDC was limited (the number of IFNgamma producing CTL was reduced by 75% compared to stimulation with peptide loaded PDC). RVV infected PDC specifically stimulated CTL but PDC were not activated. These results indicate that PDC are not ideal targets for RAd and RVV based vaccines. However, PDC induced specific CTL activation after pulsing with recombinant protein, indicating that PDC can also cross-present antigens released from surrounding infected cells. PMID- 16037639 TI - Quantitative analysis of chemotherapeutic effects in tumors using in vivo staining and correlative histology. AB - AIMS: To microscopically analyze the chemotherapeutic response of tumors using in vivo staining based on an annexinV-Cy5.5 probe and independently asses their apoptotic count using quantitative histological analysis. METHODS: Lewis Lung Carcinomas cells, that are sensitive (CS-LLC) and resistant (CR-LLC) to chemotherapy were implanted in nude mice and grown to tumours. Mice were treated with cyclophosphamide and injected with a Cy5.5-annexinV fluorescent probe. In vivo imaging was performed using Fluorescence Molecular Tomography. Subsequently tumours were excised and prepared for histology. The histological tumour sections were stained for apoptosis using a terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay. A minimum of ten tissue sections were analyzed per tumour for apoptosis quantification by TUNEL staining and corresponding Cy5.5 distribution. RESULTS: We detected higher levels of apoptosis and corresponding higher levels of Cy5.5 fluorescence in the CS-LLC vs. the CR-LLC tumours. The cell count rate on CS-LLC sections over CR-LLC was found to be approximately 2 :1 where the corresponding area observed on Cy5.5 distribution measurements revealed a approximately 1.7 :1 ratio of CS-LLC over CR-LLC. These observations are consistent with the higher apoptotic index expected from the CS-LLC cell line. CONCLUSIONS: Quantitative analysis of histological slices revealed higher fluorescence and higher apoptotic count in the CS-LLC tumour images compared to the CR-LLC tumour images. These observations demonstrate that the annexinV-Cy5.5 probe sensed the chemotherapeutic effect of cyclophospamide and further confirmed in vivo FMT measurements. PMID- 16037640 TI - Chromosomal changes in relation to clinical outcome in larynx and pharynx squamous cell carcinoma. AB - Invasive head and neck squamous carcinomas are among the cytogenetically most complex tumors. Perhaps for this reason, there is little consensus on the prognostic value of specific chromosomal aberrations. Here we present results of CGH analysis of 56 clinically well-characterized set of head and neck cancers, consisting of larynx and pharynx only. The aim was to find possible associations with clinical outcome. The major chromosome arms showing gains were (in decreasing order): 3q, 7q, 8q, 5p, 11q13, 17q and 18p, and losses occurred at 3p, 11qter, 4p, 18q, and 5q. The segments most frequently amplified were 3q26-qter, 11q13, 11q22, 3q12-13, 18p11.3, 18q11.2 and 8q24.3. Tumors with stages III and IV, and lymph node positive tumors had a worse clinical outcome. Surprisingly, no specific chromosomal abnormality correlated with disease-free survival. The only aberration that correlated to one of the clinico-pathological parameters was amplification 11q13, that occurred solely in lymph node positive, stage IV tumors. However 11q13 amplification did not correlate with disease-free survival. These results seem to indicate that genetic alterations at the level of chromosomes have limited prognostic value in patients with invasive larynx and pharynx squamous cell carcinomas. PMID- 16037641 TI - Spontaneous apoptosis in chronic lymphocytic leukemia is not an independent prognostic factor for stability of disease when compared with combined AgNOR and TTM scores. PMID- 16037642 TI - [General thoughts about use of plants in food, supplements and in phytotherapy]. AB - Herbal products on the market are evaluated by health authorities to decide whether they are drugs to all intents and purposes, and consequently they have to conform to related laws, or whether they should be regarded as dietary supplements and hence marketed freely or with notification of label to competent Authorities. The question appears to be very complicated because only a limited number of plants are utilized in phytotherapy, whereas most of them are borderline between therapeutic and physiological activity. The article summarizes the main national and community legislation on dietary supplements and herbal products. PMID- 16037643 TI - [Biologically-active phytochemicals in vegetable food]. AB - Phytochemicals constitute a heterogeneous group of substances and evidence for their role in the protective effect on human health, when their dietary intake is significant, is emerging. These compounds have biological properties as antioxidant activity, modulation of detoxification enzymes, stimulation of the immune system, decrease of platelet aggregation and modulation of hormone metabolism. Glucosinolates and widely group of polyphenols, including the main category of flavonoids, are examined. Since their weak estrogenic activity is of interest at present, isoflavones are discussed at length. Numerous factors affect the phytochemical content and availability in plants; these factors include environmental conditions and industrial or domestic processing. The bioavailability of phytochemicals can be influenced by intrinsic factors in food and/or in human, in general the substances are little adsorbed, largely metabolized and rapidly eliminated. For this reason it is advisable that their consumption is constant in the time to maintain high concentrations of metabolites in the blood. PMID- 16037644 TI - [Medicinal plants in phytotherapy]. AB - Plant kingdom has a wider biochemical diversity than animals and at least four fifths of secondary metabolites come from vegetable world. This is probably due to the link between soil and plants, therefore these have to develop numerous adaptation mechanisms. To date, about 40% of modern monomolecular drugs derives directly or indirectly from plants. The phytotherapy today constitutes the most popular medical practice of complementary medicine, and in many countries its increase is continuous. Some vegetable preparations are marked as pharmaceutical regulation. However in the most cases, in absence of clinical tests, the empiric experience matured in a long period can be considered acceptable testimony of their efficacy. The vegetable products, that are used to cure smaller indications, are based on the Directive 2004/24/EC of March 2004. PMID- 16037645 TI - [Pharmacological considerations on herbal medicine use]. AB - Herbal products are considered dietary supplements from a regulatory point of view. If we apply the basic principles of pharmacology to herbal medicines they can be classified into three groups: 1) products whose efficacy has been demonstrated: their active principles are known and the therapeutic doses are established; 2) herbal products whose efficacy is probable, but not clearly demonstrated: they contain pharmacologically active substances that are used to standardize the products. Their therapeutic dose is difficult to establish; 3) products with uncertain efficacy but with a long lasting traditional use: they can be useful for treating minor disorders, but should be employed exactly as in the traditional medicine. Most of the herbal products at present classified as herbal supplements fall into the second and third group: it is apparent that they cannot be considered herbal supplements. PMID- 16037647 TI - [Herbal supplements in sports: use and abuse]. AB - The use of natural supplements, included herbal supplements, by athletes has become an habit which often lacks any valid scientific rationale. It appears evident that this habit may entail health risks (including more or less serious adverse effects), consequent either: 1) to the pharmacodynamic effects of the drugs at high doses; or 2) to the occurrence of accumulation especially when their administration is not justified by a reduced synthesis or an increased demand; or 3) to the occurrence of intolerance; or, finally, 4) to the presence of unlabelled ingredients. The abuse of this kind of products always entails risks to the consumer, not only to the elite athlete, that can incur an adverse analytical finding on the occasion of anti-doping tests, but also to the amateur sportsman, for the possible occurrence of adverse drug reactions (ADR). PMID- 16037646 TI - [Clinical phytotherapy: opportunities and problematics]. AB - In every country traditional medicines find foundation in magical or religious beliefs, or popular experience. The World Health Organization is engaged to establish guidelines for methodology of clinical research and the appraisal of effectiveness of traditional medicine. In Italy it has been adopted the term non conventional medicines in order to group one series of therapies and medicines which are radically different one from the other. Phytotherapy uses medicinal plants prepared by means of adapted extractive technologies and proper pharmacological preparations, purified and standardized in their chemical principles. As for any shape of therapy there are possible side effects, contraindications, pharmacological interactions. It is deemed necessary that a basic knowledge of the pharmacological aspects of phytotherapy be included in the regular Course of Medicine. PMID- 16037648 TI - [Pilot study for the surveillance of adverse reactions to herbal preparations and dietary supplements]. AB - Use of herbal medicine is increasing in Italy. Herbal preparations are generally used as self-medication, often in the wrong belief that "natural" can be equated with "harmless". Whereas use of these products has been associated with serious adverse events, due to the quality of the raw materials used, to interactions with synthetic drugs or to their use in particular conditions, such as pregnancy or lactation. To monitor herbal preparation use in the population is necessary in order to point out possible adverse reactions. This was the objective of this pilot study, together with the urgency to inform health personnel and patients on the correct use of these products. After this experience we strongly believe that a system to assess the safety of herbal preparations is needed. PMID- 16037649 TI - [Quality control in herbal supplements]. AB - Quality and safety of food and herbal supplements are the result of a whole of different elements as good manufacturing practice and process control. The process control must be active and able to individuate and correct all possible hazards. The main and most utilized instrument is the hazard analysis critical control point (HACCP) system the correct application of which can guarantee the safety of the product. Herbal supplements need, in addition to standard quality control, a set of checks to assure the harmlessness and safety of the plants used. PMID- 16037650 TI - [Herbal drugs: from traditional use to regulation]. AB - Herbal preparations have been used for centuries as the main therapeutic means. In Italy there is an ancient tradition of using herbal remedies, which became extremely important from the 16th to the 18th century. Nowadays multinational companies invest great resources on herbal drugs and preparations. This article focuses on herbal medicines, herbal products, and food supplements. Moreover the European legislation on traditional medicinal plants and food supplements is analysed and discussed. PMID- 16037651 TI - [Herbal food supplements]. AB - Dietary supplements are increasingly popular. Supplements include substances such as vitamins, minerals, amino acids, essential fatty acids, fibers and botanicals. Directive 2002/46/EC foresees the use as food supplements of vitamins and minerals listed in the Annexes to the Directive. The Directive challenged the importance of a European regulatory framework for this products to be adopted by the member states. To date herbal products without any therapeutic activity have been included in food supplements. This review summarizes the main legislations on food supplements focusing on herbal products or derivatives. PMID- 16037652 TI - [The characteristics of Alzheimer's Disease Units in relation to neuropsychological tests]. AB - The Cronos Project is a post-marketing surveillance study implemented by the Italian Ministry of Health and the National Institute of Health whose main objectives are to characterise the population of Alzheimer's disease patients treated with acetylcholinesterase inhibitors and monitor effectiveness and drug safety in the field practice. In this project 503 Alzheimer's disease units were activated located throughout the country. The characteristics of these Alzheimer's disease units are presented for setting (territorial, university, hospital, extra-hospital), health personnel employed, examinations offered (CT and MRI scans and laboratory tests), counselling activities and relationship with caregiver associations in relation to neuropsychological tests. PMID- 16037653 TI - [Use of neuropsychological tests in the Cronos Project]. AB - Within the Cronos Project, 503 Alzheimer's disease units were activated throughout the country. In June/July 2002 a questionnaire was sent to all Alzheimer's disease units to collect data on neuropsychological tests utilised in the diagnostic process of dementias. Only 196 of Alzheimer's disease units that have responded to the questionnaire (196/392) declare to utilize neuropsychological test while 97.8% use the mini mental state examination. This paper aims to collect, for the first time in Italy, data on use of the neuropsychological tests in the specialist units on dementia and to propose a discussion on the reliability of neuropsychological instruments. PMID- 16037654 TI - [Dementias: mostly clinico-neuropsychological or mostly psychometric diagnosis?]. AB - In this paper many critical reflections on diagnosis management of patients affected by dementia have been reported. In particular, the importance of clinico neuropsychological evaluation of patients with respect to management prevalently based on neuropsychological tests is underlined. In this view a periodic diagnostic exercise with the objective of improving the diagnostic approach of dementias has been proposed to the Alzheimer's disease units (UVA) included in the Cronos Project. PMID- 16037655 TI - [Variability of epidemiological measures in mild cognitive impairment and dementia]. AB - Variability in occurrence estimates is one of the basic features of the epidemiology of dementia and mild cognitive impairment (MCI). This review will cover two levels of variability that affect epidemiological research on dementia and MCI: the conceptual and the operational level. More specifically, it is highlighted how the lack of a precise definition of MCI leads to a greater variability in the occurrence estimates of this condition, when compared to dementia. Variability will decrease only when more precise criteria and aims of the concept "MCI" will be specified. PMID- 16037656 TI - [Variability of efficacy measures in Alzheimer's disease]. AB - The efficacy of medical interventions is their capacity of inducing positive modifications of the natural history of diseases. The natural history of dementia is marked by specific events related to the cognitive and functional decline, but their occurrence is poorly predictable in individual patients being highly variable from patient to patient. For this reason it is difficult that the modest efficacy of available interventions for dementia, or their entity measured by clinical scales, may be perceived in clinical practice or in observational studies. Moreover in randomized clinical studies, the effect of this variability, in analogy to misclassification of exposition and/or disease in case control or cohort epidemiological studies, is that of an underestimation of the true efficacy of interventions. PMID- 16037657 TI - [In-hospital management of victims of chemical weapons of mass destruction]. AB - Emergency situations caused by chemical weapons of mass destruction add a new dimension of risk to those handling and treating casualties. The fundamental difference between a hazardous materials incident and conventional emergencies is the potential for risk from contamination to health care professionals, patients, equipment and facilities of the Emergency Department. Accurate and specific guidance is needed to describe the procedures to be followed by emergency medical personnel to safely care for a patient, as well as to protect equipment and people. This review is designed to familiarize readers with the concepts, terminology and key operational considerations that affect the in-hospital management of incidents by chemical weapons. PMID- 16037658 TI - [Creutzfeldt-Jakob disease and other human forms of transmissible spongiform encephalopathy in Italy: a mortality study carried out from different data sources]. AB - Creutzfeldt-Jakob Disease (CJD) is a rare pathology (about 1 case per million) but it has a great importance for Public Health; the Italian National CJD register has been established in the Istituto Superiore di Sanita (ISS) since 1993, and epidemiological studies on CJD have been carried out as well. This paper reports a mortality study carried out comparing and integrating data from the two available sources: the National CJD Register and the Italian Data Base on Mortality, processed by the ISS Statistics Unit, on the data collected by the Italian Census Bureau (ISTAT). The study allowed to estimate: the underreporting of CJD mortality to both sources, the misclassification of ISTAT data and the integrated mortality rates from CJD in Italy: 1.58 per million persons aged 25 or more, average rate during the period 1993-1999. PMID- 16037659 TI - [Youth and HIV: information campaigns by and for adolescents]. AB - This article describes a pilot project carried out in collaboration between the Istituto Superiore di Sanita and the Azienda Sanitaria Locale Roma E without any specific funding. The aim was to take a group of adolescents and provide them with a grounding in both HIV/AIDS infection and social communication, and with the instruments necessary to develop an informative campaign with other adolescents as the target group. The project was divided into three phases: sessions for raising levels of HIV/AIDS information and awareness involving 702 secondary school students; workshops to provide 120 selected students with communication and advertising know-how, to allow them to develop an HIV/AIDS infection information campaign targeted at their peers; a final event for the presentation of the students' findings. Prevention was the focus of the adolescents' resulting products, with particular attention to condoms as means of protection. The target population was judged as best influenced by channels such as posters and television ads, and the resulting messages were cartoon based, both ironic and fun yet accompanied by strong and direct statements designed to shock the viewer. The methods used in the project turned out to be particularly suitable for giving importance to the input of the participants who went from being publicity targets to developers. PMID- 16037660 TI - Quantitative assay of total dsDNA with PicoGreen reagent and real-time fluorescent detection. AB - We describe a quantitative assay of dsDNA based on real-time PCR measurement of fluorescence due to the interaction of PicoGreen dye with dsDNA. An aliquot of 1 to 5 ml of the sample is mixed with 45 ml of diluted PicoGreen reagent within an optical PCR tube. This is placed into the real-time apparatus set to read SYBR Green I dye at the end of three cycles of 94 degrees C for 30 s and 65 degrees C for 30 s. The averaged fluorescence value is converted into DNA amount using a calibration curve prepared with lambda-DNA standard. The calibration curve has a dynamic linear range from 0.20 to 50 ng and a standard deviation variability below 5.0%. In conclusion, this method allows reliable determinations on minimal amounts of DNA from biological samples and PCR products in clinical applications of molecular biology. PMID- 16037661 TI - Combining time-resolved and single-phase 3D techniques in contrast-enhanced carotid MR angiography. AB - We established an easy-to-use technique for performing contrast-enhanced carotid MR angiography (MRA) with a commercial scanner. Twenty-three patients with suspected carotid or vertebral arterial lesions were prospectively studied. Two techniques were applied in the study. After performing sagittal time-resolved acquisitions, we undertook a coronal single-phase 3D acquisition, in which the injection timing was estimated from the preceding images. In each case, we obtained multidirectional images with sufficient venous suppression. The combined use of time-resolved and single-phase 3D MRA is a feasible technique for obtaining selective arterial images without the use of special applications or hardware. PMID- 16037662 TI - In vitro 1H-NMR spectroscopic analysis of metabolites in fast- and slow-twitch muscles of young rats. AB - The lactate (LAC), creatine (CRN), taurine (TAU), anserine (ANS) and carnosine (CAR) content of the masseter muscles (MM), long extensor muscles of digits (EDL) and soleus muscles (SOL) of young rats were determined using in vitro 1H-NMR spectroscopy to assess the significance of CRN, TAU, ANS and CAR in these muscles. The muscles of Wistar rats at the ages of 6, 12 and 18 weeks were dissected after decapitation and used for the metabolite analyses. The LAC and CAR content of all muscle groups showed no age dependence. The CRN content was increased age-dependently in MM but not in EDL or SOL. The LAC and CRN content was higher in MM and EDL (fast-twitch) than in SOL (slow-twitch) (P<0.01-0.001 at 18 weeks). A significant positive correlation existed between the LAC and CRN content (P<0.00001, r=0.80), suggesting that the CRN content reflects the capacity of the anaerobic glycolysis of the individual muscles. The TAU content was higher in SOL and MM than in EDL (P<0.05) and showed an approximately 1.5 fold increase with age in all three muscle groups. The ANS content was higher in EDL than in SOL and MM (P<0.001), and showed an approximately threefold increase with age in all three muscle groups. The ANS content positively correlated with the LAC content (P<0.001, r=0.41), and the chemical shift of the imidazole proton in ANS showed a correlation with the LAC content (P<0.0001, r>0.76), indicating that ANS would buffer the pH change produced by LAC. These results suggest that 1H-NMR spectroscopy would provide an adjunct method of assessing the muscle types and their development. PMID- 16037663 TI - Image-based diagnosis of Alzheimer-type dementia: measurements of hippocampal and ventricular areas in MR images. AB - We studied the possibility of objective diagnosis of Alzheimer-type dementia through radiological findings by evaluating cerebral and hippocampal atrophy in magnetic resonance images of patients with dementia. A statistically significant difference was seen in the hippocampus area index (HAI) and the ventricle area index (VAI) of healthy elderly individuals. Discriminant analysis with these two indexes demonstrated a sensitivity of 90.2% and a specificity of 81.8%, with an overall accuracy of 87.7%. Multiple discriminant analysis with VAI and HAI by stage of dementia showed an overall accuracy of 61.6% for discrimination of four stages. These results indicate that quantitative magnetic resonance imaging measurements can be used as an objective diagnostic technique for Alzheimer-type dementia. PMID- 16037664 TI - Manipulo-spatial processing of ideographic characters in left-handers: observation in fMRI. AB - It has been suggested that left-handers have a cerebral ambilaterality for language representation. Specifically, the use of the right hand for writing may have a specific effect on the cerebral organization in left-handers. In an investigation of the relationship between motor and visual language procedures, functional magnetic resonance imaging at three tesla was conducted during stroke counting of kanji (Japanese ideographic characters) in six left-handers who usually write with their right hand. Two types of stimulus presentation, phonography-displayed and kanji-displayed, were employed to examine the different neural pathways used for processing kanji. Each stimulus presentation involved two motor conditions: one allowed finger movements for tracing the characters, while the other disallowed finger movements. The tasks induced activation in the primary motor area, the premotor area, the supplementary motor area, and the anterior cingulate gyrus as well as the parietal and occipital lobes (Brodmann's area 7/39/19). The activated areas in both the movement-allowed and movement disallowed conditions were almost identical except for the primary motor area. These results clearly contrasted with those of a previous study of right-handers which showed that right-handed volunteers demonstrated decreased activation in the premotor area and the dorsal pathway during the movement-allowed condition. This discrepancy may be attributable to a difference in cerebral organization for language processing. Specifically, in left-handers, the visuospatial procedure for kanji and the motor procedure for tracing the kanji may be ambilaterally distributed in both hemispheres, whereas in right-handers these procedures may be predominantly lateralized in the left hemisphere. PMID- 16037665 TI - An emerging concept of biomolecular dynamics and function: applications of NMR & MRI. AB - A new concept of protein dynamics has emerged quite recently, and a crucial link between protein dynamics and function has been largely established using recent NMR techniques in the solution state. Protein structure is governed by the thermodynamic principle and may not necessarily be unique in the solution state. Enzyme catalysis, protein folding or allosteric transition occurs on the microsecond to millisecond time scale, implying that in order to prepare the specific nuclear coordinate for the electronic state transition, a protein must rearrange its nuclear coordinates substantially, and this process may generally take a long period of time almost comparable to that of protein folding. Protein coordinates optimized for the electronic reaction may form an energy state--which may be called an "excited state"--that is thermodynamically distinct from the native state. In contrast, the native state is called a "ground state." Relevant NMR techniques developed recently may also have useful application to MRI, since the critical time scale of various reactions in a living system is also around micro- to milliseconds. PMID- 16037666 TI - Dynamic MR imaging and radiotherapy. AB - Dynamic MRI has been used to improve the detection of tumors and to make differential diagnosis. Most malignant lesions show early enhancement and early washout of contrast media on dynamic MRI, but the characterization of the tumor remains unclear. Pharmacokinetic analysis of dynamic MRI can provide information about the permeability of contrast media in the tumor that may reflect the oxygen concentration of the tumor. This information may be useful in the prediction of a tumor's response to radiation therapy. PMID- 16037667 TI - Recent topics in NMR imaging and MRI. AB - NMR and NMR imaging (MRI) are finding increasing use not only in the clinical and medical fields, but also in material, physicochemical, biological, geological, industrial and environmental applications. This short review is limited to two topics: (i) new techniques and pulse sequences and their application to non clinical fields that may have clinical application; and (ii) new trends in MR contrast agents. The former topic addresses pulse sequence and data analysis; dynamics such as diffusion, flow, velocity and velocimetry; chemometrics; pharmacological agents; and chemotherapy; the latter topic addresses contrast agents (CA) sensitive to biochemical activity; CA based on water exchange; molecular interactions and stability of CA; characteristics of emerging CA; superparamagnetic CA; and macromolecular CA. PMID- 16037668 TI - A case of MELAS: hyperperfused lesions detected by non-invasive perfusion weighted MR imaging. AB - We report a case of MELAS in which lesions were detected by perfusion-weighted MR imaging. Perfusion-weighted MRI using contrast media (PWI) and FAIR (flow sensitive alternating inversion recovery), an arterial spin labeling method, clearly showed these lesions as hyperperfused areas. One of these lesions, diminished after steroid therapy, could also be detected by FAIR. PMID- 16037669 TI - MR imaging of multifocal autoimmune pancreatitis in the pancreatic head and tail: a case report. AB - Autoimmune pancreatitis, unlike alcoholic chronic pancreatitis, is a rare variant of pancreatitis. We describe the MR imaging findings in a 50-year-old man with multifocal autoimmune pancreatitis in the pancreatic head and tail. Our case demonstrates multifocal enlargement, diminished signal intensity on a T1-weighted image, and delayed enhancement with the capsule-like hypointense bands in a dynamic study. MR cholangiopancreatography is also found to be useful in evaluating the effectiveness of steroid therapy. PMID- 16037670 TI - A case of testicular malignant lymphoma with extension to the epididymis and spermatic cord. AB - Findings of magnetic resonance imaging (MRI) in a case of testicular lymphoma are presented. The tumor was lower intensity than the normal testis on T2-weighted images. Coronal images clearly showed the tumor extending to the spermatic cord. Compared with seminoma, lymphoma more often invades the epididymis and spermatic cord. MRI is a useful method for differentiating between testicular lymphoma and seminoma because it clearly shows tumor extension to the epididymis and spermatic cord. PMID- 16037671 TI - Laparoscopic surgery for inflammatory bowel disease. AB - Laparoscopic surgery has recently been gaining acceptance as an alternative approach for patients with inflammatory bowel disease. There is increasing evidence demonstrating the multiple potential benefits of laparoscopy including faster recovery, reduced costs, and lower morbidity. For patients with acute colitis, a laparoscopic subtotal colectomy and end ileostomy have been shown to be feasible and safe in experienced hands. When indicated, many of these patients may be able to safely undergo a subsequent laparoscopic approach for construction of an ileo-anal pouch. Although still controversial, an elective laparoscopic restorative proctocolectomy with ileo-anal pouch anastomosis has also been shown to be feasible with functional outcomes at least similar to those obtained with an open approach. However, larger randomized series of patients are needed with longer follow-up in order to draw definite conclusions. For Crohn's disease, a laparoscopic approach is ideal for stoma creation. In addition, laparoscopic ileo colectomy is arguably the preferred approach for patients with terminal ileal disease. Some experienced laparoscopic groups have also applied laparoscopic techniques for more complicated cases with recurrent disease or disease-related complications, such as fistulous disease. Other short-term benefits of a laparoscopic approach may include a decreased incidence of ventral hernias, decreased incidence of small bowel obstruction, and faster recovery. These benefits may also have significant economic impact. In contrast to earlier reports, there is reliable evidence that conversion is not associated with a poorer outcome. A policy of starting most suitable cases laparoscopically may offer patients the potential benefits of a laparoscopic approach without increasing morbidity. PMID- 16037672 TI - Isolated resection of segment 1 of the liver. AB - Isolated resection of segment 1 is a parenchyma-sparing technique that can be used for the resection of benign or malignant tumors confined to this segment. The method of resection of segment 1 is described in relation with its complex anatomical features. Special attention is paid to prevention of damage to the hilar bile ducts and major hepatic veins during the procedure. PMID- 16037673 TI - Deceptive endoscopic appearances in ulcerative colitis. PMID- 16037674 TI - Intestinal blood loss due to GIST. PMID- 16037675 TI - Ring-reinforced prosthesis for paracolostomy hernia. AB - BACKGROUND: The reported recurrence rate after paracolostomy hernia repair, with or without mesh, varies between 47 and 100%. We developed a polypropylene mesh prosthesis with a central opening, reinforced with a polypropylene ring, that can be implanted suprafascially through a mucocutaneous incision. METHODS: The results of a consecutive series of 46 patients with paracolostomy hernia, who were implanted with this prosthesis between October 1988 and November 2000 in 12 hospitals, were analyzed. The mean follow-up was 5 (range 1-13) years. RESULTS: The early infection rate was 4.3%. Two patients were lost to follow-up. The late infection rate was 2.3%. Seven patients (15.9%) developed a recurrent parastomal hernia and were all reoperated. The prosthesis was definitively removed in 10 patients. CONCLUSION: Although implantation of this new prosthesis is not without complications, it offers the lowest reported recurrence rate until now. PMID- 16037676 TI - Primary carcinoma of the appendix - Hull series. AB - BACKGROUND: Appendiceal carcinoma (AC) is a rare entity that does not have a well defined treatment strategy. At presentation, most patients are clinically thought to have appendicitis and the diagnosis is made only by formal histology. Once the diagnosis of AC is made, patients are treated by various strategies including surgery, chemotherapy depending on nodal status of the disease. AIM: To review the Hull hospitals' experience with AC. METHODS: Between 1982 and 2002, 10 patients with primary AC were seen. The histopathology reports of all appendiceal specimens removed were traced. Follow-up was by chart review or patient follow-up as appropriate. We did not include patients with primary carcinoid tumours or secondary adenocarcinoma. RESULTS: There was an equal sex distribution. All patients underwent surgery, 3 had post-operative chemotherapy. Complete follow-up information was available with a median follow-up time of 56 months, with a range of 12-168 months. Five patients survived at least 4 years from the time of diagnosis. CONCLUSION: Long-term survival in patients with AC is possible. PMID- 16037677 TI - Catechol-O-methyltransferase Val158Met polymorphism in schizophrenia: associations with cognitive and motor impairment. AB - Cognitive and motor deficits have been proposed as markers of abnormal neurodevelopment in schizophrenia and have been associated with genetic liability. In a multicenter study involving 106 subjects, 56 with deficit schizophrenia and 50 with nondeficit schizophrenia, we tested the hypothesis that the catechol-O-methyltransferase (COMT) Val(158)Met polymorphism is associated with cognitive and motor deficits either in schizophrenia as a whole or in its deficit subtype. The COMT Val(158)Met polymorphism shared 6.6% of the executive/attention dysfunction variance in patients with schizophrenia and 15.6% of the motor impairment variance in patients with deficit schizophrenia. These results support the hypothesis that the COMT Val(158)Met polymorphism influences executive functions in schizophrenia and the neuromotor performance in the deficit subtype only. PMID- 16037678 TI - Chronic mild stress (CMS) revisited: consistency and behavioural-neurobiological concordance in the effects of CMS. AB - The chronic mild stress (CMS) model of depression has high validity but has in the past been criticized for being difficult to replicate. However, a large number of recent publications have confirmed that CMS causes behavioural changes in rodents that parallel symptoms of depression. This review summarizes studies from over sixty independent research groups that have reported decreases in reactivity to rewards, and a variety of other depression-like behaviours, in rats or mice, following exposure to CMS. Together, these changes are referred to as a 'depressive' behavioural profile. Almost every study that has examined the effects of chronic antidepressant treatment in these procedures has reported that antidepressants were effective in reversing or preventing these 'depressive' behavioural changes. (The single exception is a study in which the duration of treatment was too brief to constitute an adequate trial.) There are also a handful of reports of CMS causing significant effects in the opposite direction, termed here an 'anomalous' behavioural profile. There are six neurobiological parameters that have been studied in both 'anhedonic' and 'anomalous' animals: psychostimulant and place-conditioning effects of dopamine agonists; dopamine D2 receptor number and message; inhibition of dopamine turnover by quinpirole, and beta-adrenergic receptor binding. On all six measures, CMS caused opposite effects in animals displaying 'depressive' and 'anomalous' profiles. Thus, there is overwhelming evidence that under appropriate experimental conditions, CMS can cause antidepressant-reversible depressive-like effects in rodents; however, the 'anomalous' profile that is occasionally reported appears to be a genuine phenomenon, and these two sets of behavioural effects appear to be associated with opposite patterns of neurobiological changes. PMID- 16037679 TI - Effects of fexofenadine hydrochloride in a guinea pig model of antigen-induced rhinitis. AB - Allergic rhinitis is an inflammatory disease of the nasal mucosa, induced by histamine, leukotrienes, and other substances released from mast cells. Fexofenadine hydrochloride, the active metabolite of terfenadine, is a novel, nonsedating antiallergic drug having H1 receptor antagonistic activity. Fexofenadine is effective for the treatment of allergic rhinitis. However, its mechanism of action in attenuating nasal congestion has not yet been elucidated. Therefore, we first examined the effects of fexofenadine on a guinea pig model of antigen-induced rhinitis. We also evaluated the effects of mepyramine, zafirlukast and ramatroban in this model; these drugs are an H1 receptor antagonist, a selective leukotriene antagonist and a selective thromboxane antagonist, respectively. Rhinitis was induced by ovalbumin (OVA) instillation into the nasal cavity of animals that had been sensitized by two earlier OVA injections (s.c. and i.p.). The nasal airway resistance was measured for 45 min after the challenge. Fexofenadine hydrochloride (20 mg/kg) and terfenadine (20 mg/kg) administered orally 70 min prior to the challenge significantly inhibited (fexofenadine, p < 0.001, terfenadine, p < 0.05) the increase in nasal airway resistance. Ramatroban (30 mg/kg) administered orally 60 min prior to the challenge also significantly inhibited (p < 0.05) the increase in nasal airway resistance. In contrast, mepyramine (3 mg/kg i.v.) and zafirlukast (3 mg/kg p.o.) failed to reduce the increase in nasal airway resistance. These results suggest that thromboxane may be involved in the increase in the nasal airway resistance in this model. Accordingly, fexofenadine may reduce the increase in nasal airway resistance by inhibiting the release of chemical mediators, including thromboxane, that are involved in the increase in nasal airway resistance in this model. PMID- 16037680 TI - Biological compatibility of polymethyl methacrylate, hydrophilic acrylic and hydrophobic acrylic intraocular lenses. AB - PURPOSE: Extensive clinical investigations of the biocompatibility of different intraocular lenses (IOLs) have been made in an effort to optimize the outcome of modern cataract surgery. The aim of this study was to add animal eye experimental implantation data regarding cellular reaction on the anterior surface of IOLs. METHODS: Thirteen adult albino rabbits had phacoemulsification/aspiration of the crystalline lens followed by implantation of a posterior chamber IOL in each eye. Three types of IOLs were studied: Hydroview (Bausch and Lomb; n = 7), Acrysof (Alcon, USA; n = 7), and polymethyl methacrylate (PMMA; HOYA, Japan; n = 7). The animals were killed by intravenous pentobarbital 1, 4, or 8 weeks later. The IOLs were explanted and stained with hematoxylin and eosin, and observed under a light microscope. The shape of mouse ascites-induced macrophages on the anterior surface of the three different IOL types (Hydroview, PMMA, and Acrysof) was studied after 24 h of oven culture. RESULTS: Hydrophilic acrylic IOLs showed the highest affinity for lens epithelial cell (LEC) outgrowth, and the lowest and slowest maturation rate reaction of macrophages. PMMA IOLs showed the lowest affinity for LEC outgrowth, and the highest reaction of macrophages. Hydrophobic acrylic IOLs showed intermediate results both regarding LECs and macrophages. CONCLUSIONS: Results suggest that IOL biomaterial properties are the key factor that influences the quantity of monocytes/macrophages as well as the process of their maturation/senescence. LEC outgrowth is influenced both by the biomaterial of IOLs and by the monocyte/macrophage reaction. PMID- 16037681 TI - Tissue inhibitor of metalloproteinase 1 adenoviral gene therapy alone is equally effective in reducing restenosis as combination gene therapy in a rabbit restenosis model. AB - Neointimal formation is a common feature after angioplasty, bypass grafting and stenting. Angioplasty damages endothelium, causing pathological changes in arteries which lead to smooth muscle cell proliferation, synthesis of extracellular matrix components and eventually restenosis formation. Adenoviruses offer an efficient transgene expression in the vascular system. In this study, we compared the effects of different gene combinations. We wanted to find out whether adenoviral catheter-mediated delivery of an additive combination of the vascular endothelial growth factor (VEGF)-A with VEGF-C is more effective than the combination of tissue inhibitor of metalloproteinase 1 (TIMP-1) alone or with VEGF-C in a rabbit balloon denudation model. Additionally, we wanted to clarify whether the combination therapy prolongs the treatment effect. It was found that TIMP-1 alone prevents restenosis and that the combination of VEGF-A and VEGF-C has a similar effect at the 2-week time point. However, the combination of VEGF-A and VEGF-C lost the treatment effect at the 4-week time point due to the catch-up growth of neointima. On the other hand, TIMP-1 and the combination of TIMP-1 with VEGF-C still had an extended treatment effect at the 4-week time point. When considering the gene combination used in this study, it is concluded that gene therapy with adenoviral TIMP-1 alone is sufficient in reducing restenosis and that combination gene therapy does not bring any significant advantages. PMID- 16037682 TI - Intracranial arteriovenous malformations treated utilizing a linear accelerator based patient rotator or commercially available radiosurgery system. AB - PURPOSE: To report a single-institution experience with intracranial arteriovenous malformations (AVMs) treated utilizing a linear accelerator-based patient rotator (PR) or BrainLAB (BL) radiosurgery system (BrainLAB AG, Heimstetten, Germany). METHODS AND MATERIALS: Since 1989, 84 evaluable patients were treated. PR patients (n = 45) were planned/localized on the basis of biplane angiography and treated between 1989 and 2000. BL patients (n = 39) were planned/localized on the basis of CT/MRI and treated since 2000. Kaplan-Meier analyses of survival, nidus obliteration (NO), and any radiographic improvement were undertaken with Cox regression of dose and volume effects. RESULTS: No significant complication, survival, previous embolization incidence, AVM location or size differences existed between BL/PR patients. The groups differed significantly in prescribed dose (PR: 16.2 Gy, BL: 17.3 Gy, p = 0.004) and isodose (PR: 62%, BL: 79%, p < 0.0001). Estimated 2-year NO rate was 87% for BL patients, 12% for PR patients (p < 0.0001). Ultimate PR NO rate was 67% at 6 years. Dose (p = 0.037) and isodose (p = 0.014) significantly affected PR NO outcome; volume was of borderline significance (p = 0.069). No factors significantly affected BL outcome. Analyses of small (< or = 4.0 cm3), high-dose (> or = 17.0 Gy) PR patients (PR1 group) vs. BL patients still demonstrated greater NO (p = 0.04) and radiographic improvement (p = 0.0004) rates for the BL group. PR1 patients had a 76% 3-year NO rate. CONCLUSIONS: BL-based radiosurgery achieved a high NO rate, the PR method did not. Differences in outcomes between PR/BL groups may be due to localization methods or an inherent advantage with the BL system. PMID- 16037683 TI - Usefulness of an intraoperative electrophysiological navigator system for subthalamic nucleus surgery in Parkinson's disease. AB - OBJECTS: The characteristics and benefits are presented of an intraoperative neurophysiological navigator computerized system developed in our center (Columbus-Track 1.0) that helps the surgical team in neuronal identification and in strategy processes in subthalamic nucleus surgery for Parkinson's disease (PD). METHODS: The navigator consists of three assembled parts: (1) neuronal identification, based on wavelet processing, filtering and gaussian characteristics of the signal; (2) track identification, based on anatomical coincidence, somatomotor response and microstimulation quotient, and (3) strategy, coordinating correction for the next track. A retrospective comparative study was performed with 15 consecutive PD patients (30 targets) operated without the system and the next 15 consecutive patients operated with the aid of the system. With the aid of the computerized navigation system, a significant reduction in the number of tracks was observed (t = -2.503, p = 0.0015), with a mean difference of 1.2 tracks per hemisphere. A non-significant reduction in the total intervention time was also observed, with a mean difference of 20 min per hemisphere (t = -1.418, p = 0.161). CONCLUSIONS: The intraoperative computerized navigation system can aid the surgical team in better identifying the neuronal signal and in defining the optimal track to achieve the target. PMID- 16037684 TI - Study of tobacco habits and alterations in enzymatic antioxidant system in oral cancer. AB - AIM AND OBJECTIVE: Tobacco is a major etiological factor for oral cancer development, accounting 30-40% of all cancer cases in India. Tobacco consumption generates free radicals and causes oxidative damages. In order to counteract these lethal effects, normal living cells have multiple antioxidant defense systems in a cascade manner. Thus, it seems that studying biological parameters, like antioxidant enzyme system, may be helpful in risk assessment and early diagnosis of oral cancer. Therefore, we analyzed erythrocytic and tissue antioxidant enzyme activities in terms of glutathione-S-transferase (GST), glutathione reductase (GR), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and plasma thiol levels. MATERIALS AND METHODS: Study included healthy controls with no habit of tobacco (NHT, n = 25), controls with habit of tobacco (WHT, n = 31) and oral cancer patients (n = 52). All the parameters were analyzed with highly sensitive and specific spectrophotometric methods. RESULTS: Erythrocytic SOD and plasma thiol levels were significantly lower (p = 0.03), while GPx and CAT levels were higher (p = 0.017) in WHT as compared to NHT. No significant changes in GST and GR levels were observed between NHT and WHT. GST, GR, SOD and CAT activities were significantly higher (p = 0.05, p < 0.001, p = 0.003 and p < 0.001, respectively) while GPx and thiol levels were lower (p = 0.035 and p < 0.001, respectively) in oral cancer as compared to WHT. Odds ratio for erythrocytic GR, SOD, CAT and plasma thiol showed significantly higher risk of oral cancer development in WHT. Mean levels of SOD and CAT were increased, while GPx and thiol were decreased with the increase in habit duration in oral cancer. GST, GR and SOD activities were significantly higher (p = 0.0001, p = 0.005 and p = 0.005, respectively), while, CAT and thiol levels were lower (p = 0.0001 and p = 0.015, respectively) in malignant tissues as compared to adjacent normal tissues. CONCLUSION: The data revealed that evaluation of antioxidant enzyme activities and thiol levels in WHT can be helpful to identify individuals at a higher risk of oral cancer development PMID- 16037685 TI - Phase II study of an 'all-oral' regimen of capecitabine, idarubicin and cyclophosphamide for metastatic breast cancer--safety, efficacy and quality of life. AB - OBJECTIVE: Patients with metastatic breast cancer (MBC) generally have a poor prognosis. Many of these patients have a good performance status. A new all-oral regimen (XIC) was evaluated in a phase II trial. The impact of the regimen on the safety and efficacy of the drug, as well as quality of life (QOL) of the patients was assessed. PATIENTS AND METHODS: From September 2000 to September 2001, informed consent was obtained from 20 heavily pretreated women with MBC. They were placed on a 6-week cycle regimen comprising capecitabine (X; 2,000 mg/m2/day in two divided doses for 2 weeks then 1 week rest), idarubicin (I; 10 mg/m2/day, days 1, 3 and 5) and cyclophosphamide (C; 100 mg/m2/day for 2 weeks then 1 week rest). RESULTS: Toxicities were generally tolerable. One patient had grade III neutropenia, which was reversible on cessation of treatment. One patient (5%) had a complete response and 4 patients (20%) achieved partial responses, yielding an overall response rate of 25%. Eight patients (40%) had stable disease. Median time to disease progression and median survival time were 13.4 and 23.7 months, respectively. Global and physical EORTC QLQ-30 scores showed no significant decrease in QOL. CONCLUSION: This is a small-scale study. XIC was generally well tolerated and favoured by the patients. This moderately active and convenient 'all-oral' regimen deserves clinical trials at a wider scale. PMID- 16037686 TI - Clinical applications of endoscopic ultrasound to oncology. AB - Endoscopic ultrasound (EUS) is a useful imaging modality in patients with certain gastrointestinal malignancies as well as lung cancer. In many cases, EUS has been shown to have superior staging accuracy compared with other imaging techniques such as computed tomography, magnetic resonance imaging and positron emission tomography. This article will review the role of EUS in diagnosing and staging esophageal, rectal, pancreatic and lung cancers as well as subepithelial lesions. Newer therapeutic applications of EUS, namely endoscopic mucosal resection for removal of localized lesions as well as celiac plexus neurolysis for pain control, will also be discussed. PMID- 16037687 TI - Inhibitors of epidermoid growth factor receptor suppress cell growth and enhance chemosensitivity of nasopharyngeal cancer cells in vitro. AB - OBJECTIVE: Epidermoid growth factor receptor (EGFR, HER1) is overexpressed in a majority of head-and-neck cancers, including nasopharyngeal carcinoma (NPC). Although EGFR inhibitors appear to be effective for some head-and-neck cancers, their efficacy in NPC remains unclear. METHODS: The effect of EGFR-specific tyrosine kinase inhibitors, including PD153035 and ZD1839, were studied in NPC TW01, NPC-TW04, and HONE1 cell lines. The effect of combining EGFR inhibitors with cytotoxic agents was evaluated in NPC-TW04 cells. RESULTS: All three NPC cell lines expressed EGFR. PD153035 and ZD1839 inhibited the growth of NPC cells with IC50s around 10 and 20 microM, respectively. These inhibitors, however, effectively suppressed ligand-stimulated EGFR activation in NPC cells with a much lower concentration (> or =0.1 microM). The growth-suppression activity of EGFR inhibitors was closely associated with suppression of AKT phosphorylation. LY294002, a phosphatidylinositol-3 kinase (P13K)/AKT inhibitor, did suppress the growth of NPC cells. Pretreatment of EGFR inhibitors by 24 h significantly enhanced the cytotoxic effect of doxorubicin, paclitaxel, cisplatin, and 5 fluororuacil in NPC-TW04 cells. CONCLUSIONS: Our data indicate that inhibition of EGFR activation is not sufficient to induce growth inhibition in NPC cells in vitro. EGFR inhibitors may be useful adjuncts in treating NPC when combined with conventional anticancer drugs. PMID- 16037688 TI - Analysis of genetic alterations associated with DNA diploidy, aneuploidy and multiploidy in gastric cancers. AB - OBJECTIVE: Recent studies have shown a close association between DNA ploidy status (diploidy, aneuploidy and multiploidy) identified by the crypt isolation technique and specific genetic alterations in colorectal carcinomas. However, such an association has not been elucidated for gastric tumors, even though they share common genetic features with colorectal carcinomas. In the present study, we established an association between DNA ploidy status and genetic alterations in gastric cancer. METHOD: The DNA ploidy status of gastric tumors was classified as diploid, aneuploid or multiploid using the crypt isolation technique, which allows isolation of pure tumor crypt from tumor tissue. Crypt isolation combined with DNA cytometric sorting, polymerase chain reaction assay using 26 microsatellite markers and direct sequencing of the p53 gene were used to detect allelic imbalances [loss of heterozygosity (LOH) or allelic loss], microsatellite imbalance (MSI) and mutation of p53 in 54 gastric cancers (13 diploid, 12 aneuploid, 29 multiploid). RESULT: Diploid tumors showed few genetic alterations, including allelic imbalances and p53 mutations. In contrast, aneuploid tumors and multiploid tumors (in particular, aneuploid populations of multiploid tumors) exhibited multiple genetic alterations, including allelic imbalances and p53 mutations. In addition, the frequencies of genetic alterations observed in the corresponding diploid fractions of multiploid tumors were relatively higher than in diploid tumors. MSI was commonly observed in diploid, aneuploid and multiploid carcinomas. CONCLUSIONS: The present results indicate that in gastric carcinomas, diploid tumors are generally non-LOH and MSI, whereas aneuploid and multiploid tumors are associated with LOH and MSI, suggesting that the genetic profile of these carcinomas is dependent on the tumor's ploidy status. PMID- 16037689 TI - The negative regulation of red cell mass by neocytolysis: physiologic and pathophysiologic manifestations. AB - We have uncovered a physiologic process which negatively regulates the red cell mass by selectively hemolyzing young circulating red blood cells. This allows fine control of the number of circulating red blood cells under steady-state conditions and relatively rapid adaptation to new environments. Neocytolysis is initiated by a fall in erythropoietin levels, so this hormone remains the major regulator of red cell mass both with anemia and with red cell excess. Physiologic situations in which there is increased neocytolysis include the emergence of newborns from the hypoxic uterine environment and the descent of polycythemic high-altitude dwellers to sea level. The process first became apparent while investigating the mechanism of the anemia that invariably occurs after spaceflight. Astronauts experience acute central plethora on entering microgravity resulting in erythropoietin suppression and neocytolysis, but the reduced blood volume and red cell mass become suddenly maladaptive on re-entry to earth's gravity. The pathologic erythropoietin deficiency of renal disease precipitates neocytolysis, which explains the prolongation of red cell survival consistently resulting from erythropoietin therapy and points to optimally efficient erythropoietin dosing schedules. Implications should extend to a number of other physiologic and pathologic situations including polycythemias, hemolytic anemias, 'blood-doping' by elite athletes, and oxygen therapy. It is likely that erythropoietin influences endothelial cells which in turn signal reticuloendothelial phagocytes to destroy or permit the survival of young red cells marked by surface molecules. Ongoing studies to identify the molecular targets and cytokine intermediaries should facilitate detection, dissection and eventual therapeutic manipulation of the process. PMID- 16037690 TI - Characterization of novel airway submucosal gland cell models for cystic fibrosis studies. AB - Cultured airway epithelial cells are widely used in cystic fibrosis (CF) research as in vitro models that mimic the in vivo manifestations of the disease and help to define a specific cellular phenotype. Recently, a number of in vitro studies have used an airway adenocarcinoma cell line, Calu-3 that expresses submucosal gland cell features and significant levels of the wild-type CFTR mRNA and protein. We further characterized previously described CF tracheobronchial gland cell lines, CFSMEo- and 6CFSMEo- and determined that these cell lines are compound heterozygotes for the F508del and Q2X mutations, produce vestigial amounts of CFTR mRNA, and do not express detectable CFTR protein. Electrophysiologically, both cell lines are characteristically CF in that they lack cAMP-induced Cl- currents. In this study the cell lines are evaluated in the context of their role as the CF correlate to the Calu-3 cells. Together these cell systems provide defined culture systems to study the biology and pathology of CF. These airway epithelial cell lines may also be a useful negative protein control for numerous studies involving gene therapy by cDNA complementation or gene targeting. PMID- 16037691 TI - ClC-3-independent sensitivity of apoptosis to Cl- channel blockers in mouse cardiomyocytes. AB - It has been shown that Cl-/HCO3- exchangers and Cl- channels, both of which are sensitive to stilbene derivatives, have essential roles in the mechanism of apoptosis induction. Staurosporine-induced apoptosis in neonatal mouse cardiomyocytes was prevented by a stilbene derivative, DIDS. To clarify whether Cl-/HCO3- exchangers or Cl- channels are targets of DIDS and whether ClC-3 is involved in the apoptotic process, staurosporine-induced reduction of cell viability, DNA laddering and caspase-3 activation were examined in cultured mouse ventricular myocytes derived from wild-type and ClC-3-deficient mice. Staurosporine-induced apoptosis and its DIDS sensitivity in ambient HCO3(-)-free conditions in which operation of Cl-/HCO3- exchangers is minimized were indistinguishable from when HCO3- was present. Apoptosis was also prevented by application of a non-stilbene-derivative Cl- channel blocker, NPPB, which cannot block Cl-/HCO3- exchangers. Cardiomyocytes derived from ClC-3-deficient mice similarly underwent apoptosis after exposure to staurosporine; moreover, apoptosis was prevented by application of DIDS or NPPB. Thus, we conclude that in cardiomyocytes, apoptosis is critically dependent on operation not of Cl-/HCO3- exchangers but of Cl- channels which are distinct from ClC-3. PMID- 16037692 TI - Effect of peroxynitrite on passive K+ transport in human red blood cells. AB - Peroxynitrite is generated in vivo by the reaction between nitric oxide, from endothelial and other cells, and the superoxide anion. It is therefore pertinent to examine its effects on the membrane permeability of red blood cells. Treatment of human red blood cells with peroxynitrite (nominally 1 mM) markedly stimulated passive K+ permeability. The main effect was on a Cl(-)-independent K+ pathway, which remains unidentified. Although K+-Cl- cotransport (KCC) was stimulated, this was dependent on saline composition, being inhibited by physiological levels of glucose (IC50 4 mM), and also by sucrose and MOPS. Effects on the Cl(-) independent K+ pathway were less dependent on saline composition, and were not inhibited by amiloride, ethylisopropylamiloride, dimethylamiloride or gadolinium. Na+-K+-2Cl- cotransporter was inhibited whilst there was little effect on the Gardos channel (Ca2+-activated K+ channel). Peroxynitrite was markedly more effective in oxygenated cells than deoxygenated ones. Treatment with peroxynitrite per se did not affect initial cell volume. Anisotonic swelling modestly increased the Cl(-)-independent K+ influx, but did not affect peroxynitrite-stimulated KCC. Decreasing extracellular pH from 7.4 to 7.2 or 7.0 increased KCC stimulation, whilst the Cl(-)-independent component of K+ transport was lowest at pH 7.2. Finally, protein phosphatase inhibition with calyculin A (100 nM) inhibited KCC, implying that, as with other KCC stimuli, peroxynitrite acts via decreased protein phosphorylation; pre-treatment with calyculin A also inhibited the Cl(-)-independent component of K+ transport. These findings are relevant to the actions of peroxynitrite in vivo. PMID- 16037693 TI - Inhibition of glutamine synthetase triggers apoptosis in asparaginase-resistant cells. AB - The resistance to L-asparaginase (ASNase) has been associated to the overexpression of asparagine synthetase (AS), although the role played by other metabolic adaptations has not been yet defined. Both in ASNase-sensitive Jensen rat sarcoma cells and in ARJ cells, their ASNase-resistant counterparts endowed with a five-fold increased AS activity, ASNase treatment rapidly depletes intracellular asparagine. Under these conditions, cell glutamine is also severely reduced and the activity of glutamine synthetase (GS) is very low. After 24 h of treatment, while sensitive cells have undergone massive apoptosis, ARJ cells exhibit a marked increase in GS activity, associated with overexpression of GS protein but not of GS mRNA, and a partial restoration of glutamine and asparagine. However, when ARJ cells are treated with both ASNase and L-methionine sulfoximine (MSO), an inhibitor of GS, no restoration of cell amino acids occurs and the cell population undergoes a typical apoptosis. No toxicity is observed upon MSO treatment in the absence of ASNase. The effects of MSO are not referable to depletion of cell glutathione or inhibition of AS. These findings indicate that, in the presence of ASNase, the inhibition of GS triggers apoptosis. GS may thus constitute a target for the suppression of ASNase-resistant phenotypes. PMID- 16037694 TI - Tongue-palate contact during selected vowels in children with cleft palate. AB - This study reports tongue-palate contact recorded using electropalatography (EPG) during five vowels /i/, /theta/, /I/, /o/ and /backwards c/ spoken by school-aged children with cleft palate and a group of normal speakers. All the children had articulation disorders affecting consonants but none had obvious vowel errors. Two measures were taken from the EPG data at the temporal midpoint of the vowels. The first identified the percentage of vowels produced with complete coronal constriction and the second calculated amount of contact. The results showed that children with cleft palate frequently produced the high vowel /i/ with complete constriction, with 40% of /i/ targets articulated in this way. There were lower percentages for /theta/ and /I/ and no complete constrictions during the lower vowels /o/ and /backwards c/. None of the normal speakers produced any vowels with complete constriction. In terms of amount of contact, the vowels ranked /i/>/theta/>/I/>/o/>/backwards c/, with /i/ having the most and /backwards c/ the least contact. Although this ranking held for both groups, the cleft group had more contact than normal speakers, especially during high vowels. Complete constriction is viewed as a clinically relevant phenomenon that blocks oral airflow and as a result increases nasal airflow during vowels. PMID- 16037695 TI - Stuttering frequency on meaningful and nonmeaningful words in adults who stutter. AB - Previous research has suggested that word meaning can influence the loci and frequency of stuttering moments. Based on this proposition, it was hypothesized that people who stutter will exhibit a larger proportion of stuttering moments on meaningful words when compared to nonmeaningful or nonsense words. In order to test this hypothesis, stuttering frequency was examined among 9 English-speaking adults who stutter as they read a total of 126 words that were either meaningful or nonmeaningful in nature. The results indicated that approximately 117% more stuttering moments were recorded on nonmeaningful words when compared to meaningful words and this difference was statistically significant (p=0.01). These findings challenge the notion that word meaning can be considered as a reliable predictor of stuttering frequency. PMID- 16037696 TI - [Laser measuring device for phonation]. AB - INTRODUCTION: Clinical examinations with short-interval, color-filtered double flash stroboscopy enable us to estimate the movements of the vocal fold edges during phonation. So-called 'displacement bands' show the degree of the vibratory amplitude in these images. Using the two-point light projection method the displacement bands can be measured with very high accuracy. A combination of these two methods was used for velocity measurements of the vocal fold movements within the phonatory cycle. The aim of the study was to explore the influence of change in sound pressure level (SPL) and fundamental frequency (F0) on the velocity of the horizontal amplitude of the vocal folds during phonation. MATERIAL AND METHODS: 40 healthy volunteers (31 women, 9 men, average age 24.8 years) were examined. During the special videostroboscopy with short-interval, color-filtered double flashes laser spots were projected onto the vocal folds by an endoscopic two-point light projection device. The subjects had to change their SPL and F0 following a test protocol. During phonation they had to produce tones in low, middle and high chest voice as well as in falsetto register. Each subject was asked to do this in a soft, a modal, a loud and a very loud manner. Images of the phonatory cycle showing the vocal folds immediately before collision were measured separately at three different positions. About 15,000 single measurements were evaluated. RESULTS: The measured velocity of the horizontal amplitude during the closing phase of the vocal folds in chest voice was 30-160 cm/s. An increase in SPL resulted in an increasing velocity of the displacement bands. A change in F0 did not lead to statistically relevant changes in the measured velocity. CONCLUSION: A combination of short-interval, color-filtered double-flash stroboscopy and a two-point light projection method enables measurements of the velocity of vocal fold movements. The SPL is the important factor for the velocity change within the phonatory cycle. F0 seems not to covary with the velocity of the horizontal amplitude during the closing phase of the phonatory cycle. PMID- 16037697 TI - Copper in developmental stuttering. AB - It has previously been reported that men with developmental stuttering showed reduced concentration of copper in the blood, and a negative correlation between the copper level and the severity of stuttering. Disorders of copper metabolism may result in dysfunction of the basal ganglia system and dystonia, a motor disorder sharing some traits of stuttering. It has been shown that copper ions affect the dopamine and the GABA systems. With this background we investigated the plasma level of copper, the copper binding protein ceruloplasmin, and the estimated level of free copper in stuttering adults. Sixteen men with developmental stuttering were compared with 16 men without speech problems. The samples were assayed in one batch in a pseudorandom and counterbalanced order. No significant differences were found between stuttering men and the control group in any of the biological variables, and no negative correlation between copper and the general severity of stuttering was shown. On the contrary, an explorative analysis resulted in a positive correlation between high plasma copper and superfluous muscular activity during stuttering (r=0.51, p=0.04). This result indicates that there is no relation between developmental stuttering and low plasma copper in the main population of stuttering adults. PMID- 16037698 TI - Vocal tract area functions to assist in determining voiced epochs in esophageal speech. AB - Esophageal talkers may have reduced intelligibility due to both time domain and frequency domain variability. The unpredictable nature of esophageal speech can cause problems when automatic procedures are used in applications such as long distance telephone messages. The current study compared a standard coding algorithm (LPC-10e) with a novel approach to determining voiced periods (vocal tract area functions) in the speech of esophageal talkers. The results of the study showed that the sentences synthesized with the vocal tract area function algorithm were more intelligible than those synthesized with the standard LPC-10e algorithm. Supplemental information, such as vocal tract area functions, may be useful in determining voiced epochs when variability in vocal parameters is high. PMID- 16037699 TI - Significance of testicular microlithiasis. AB - INTRODUCTION: Testicular microlithiasis is an uncommon condition characterized by calcifications within the seminiferous tubules. The true prevalence in a normal population has not been defined. METHODS: A review of the literature with emphasis on the connection between testicular microlithiasis and testicular malignancy was carried out. RESULTS: Testicular microlithiasis is associated with different testicular pathologies, including testicular cancer. However, a direct causative connection between testicular microlithiasis and testicular pathologies is not supported by the literature. CONCLUSIONS: Patients with testicular microlithiasis should be followed up regularly. Further investigations concerning the etiology of testicular microlithiasis remain to be done. PMID- 16037700 TI - Endoscopic enucleation of leiomyoma of the bladder. AB - A case of benign leiomyoma of the urinary bladder was discovered incidentally and treated successfully with endoscopic enucleation. We describe the technique of this new surgical approach and discuss its advantages. We recommend this minimally invasive approach over open surgical resection. PMID- 16037701 TI - Prognostic factors in a recent series of patients treated with radical cystectomy for bladder cancer. AB - OBJECTIVE: To identify the clinical and pathological prognostic factors in a homogeneous series of patients with bladder cancer who had undergone radical cystectomy in the late 1990s. MATERIALS AND METHODS: We retrospectively analyzed the clinical data of 156 patients who had undergone radical cystectomy and iliac obturator lymphadenectomy for bladder carcinoma at our department between 1995 and 2001. RESULTS: The mean follow-up was 39.71 +/- 26.2 months. The 5-year overall and cancer-specific survival rates were 47.2 and 54.7%, respectively. Upper urinary tract obstruction (p = 0.03), clinical stage of both the primary tumor (p = 0.0001) and loco-regional lymph nodes (p = 0.04), pathological stage (2002 TNM) of the primary tumor (p < 0.0001), pathological loco-regional lymph node involvement (p < 0.0001), and vascular embolization (p = 0.005) were significant on univariate analysis. Pathological lymph node involvement (p = 0.001) and both pathological (p = 0.022) and clinical stages of the primary tumor (p = 0.002) turned out to be independent predictors of cancer-specific survival. CONCLUSION: Pathological lymph node involvement, clinical and pathological stage of the primary tumor were the cancer-specific, survival-independent, predictors in our series. Our multivariate analysis data identified pT3-4 and pN+ patients as those with the worst prognosis. PMID- 16037702 TI - Urothelial cancer of the renal pelvis in percutaneous nephrolithotomy patients. AB - INTRODUCTION: The association between staghorn calculus of the kidney long standing and urothelial tumors of the renal pelvis is well documented. We describe 3 patients with urothelial cancer in a kidney who underwent percutaneous nephrolithotomy (PCNL). PATIENTS AND METHODS: Retrospective analysis of our series of more than 500 PCNLs revealed 3 patients in whom urothelial cancer of the renal pelvis was diagnosed during or following percutaneous stone removal. Preoperative assessment included IVP, renal sonography and renal scan using DTPA in all 3 patients, and CT in 2 patients. Imaging did not raise the suspicion of a neoplastic lesion. RESULTS: All patients had a long history of urolithiasis and urinary infections. No patient was diagnosed preoperatively. One patient was diagnosed postoperatively, when a CT demonstrated a renal lesion. The second patient underwent simple nephrectomy due to a non-functioning kidney, and the tumor was found on pathological analysis. In the third patient, biopsies were taken from a suspicious-looking tissue in the renal pelvis during the PCNL session. All patients had transitional cell carcinoma: 1 associated with sarcomatoid features and 1 with squamous carcinoma. They all died from metastatic disease 2-19 months after the diagnosis of urothelial cancer. CONCLUSIONS: The preoperative diagnosis of urothelial cancer in patients with staghorn stones is difficult due to the existing stone and inflammation. Since the prognosis of urothelial cancer is extremely poor, biopsies of the renal pelvis, obtained directly through the nephroscope during the PCNL session, may be the only key for early diagnosis and treatment. A high index of suspicion should be raised when patients suffering from infected staghorn calculi are encountered, and such intraoperative biopsies should be considered. PMID- 16037703 TI - Effect of different sized transurethral catheters on pressure-flow studies in women with lower urinary tract symptoms. AB - INTRODUCTION: The study evaluates the impact of three different sized (4.5-, 6- and 7-Fr) catheters on pressure-flow studies in women undergoing urodynamic evaluation for lower urinary tract symptoms. MATERIAL AND METHODS: 60 women referred for the evaluation of lower urinary tract symptoms were enrolled in this randomized controlled study. Patients were divided into two groups (A and B) of 30 women each. The patients underwent non-invasive free-flow uroflowmetry with determination of post-void residual urine volume (PVR) and two consecutive pressure-flow studies using two different transurethral catheters. In group A the two consecutive pressure-flow studies were performed using a 4.5-Fr catheter once and a 6-Fr catheter once; in group B the two consecutive pressure-flow studies were performed using a 4.5-Fr catheter once and a 7-Fr catheter once. Patients were also randomized for the consecutive order in which the two different transurethral catheters were used. RESULTS: The pressure-flow parameters were significantly different from the equivalent free-flow findings. The maximum and average flow rate in all pressure-flow studies performed were significantly lower than the equivalent free-flow parameters and the flow time was significantly longer for all pressure-flow versus free-flow studies. Furthermore, there was a significantly larger PVR for pressure-flow than for free-flow measurements. There was no significant difference in maximum flow rate, average flow rate and flow time between 4.5- and 6-Fr pressure-flow studies (group A). However, there was a statistically significant difference between 4.5- and 7-Fr pressure-flow studies (group B) in those uroflowmetry parameters. Detrusor pressure at maximum flow (Pdet. Qmax) and maximum detrusor pressure (P(det. max)) in group A did not show statistically significant differences between 4.5- and 6-Fr pressure-flow studies whereas in group B Pdet. Qmax and P(det. max) were significantly different between 4.5- and 7-Fr pressure-flow studies. CONCLUSIONS: A 4.5-, 6- or 7-Fr transurethral catheter may obstruct micturition changing uroflowmetry parameters. A statistically significant difference was found in all but PVR parameters particularly in all detrusor parameters between 4.5- vs. 7-Fr whereas no statistically significant difference was found in all but PVR parameters between 4.5- vs. 6-Fr. We can thus state that pressure-flow findings may be influenced by different sized transurethral catheters. A misinterpretation of pressure-flow findings may have clinical implications on establishing an accurate diagnosis. PMID- 16037704 TI - Removal of catheter at midnight versus early morning: the patients' perspective. AB - BACKGROUND: With an increasing role of the patient as a partner in making a combined decision in care plan goals, it is important to identify the patient's perspective of the experience of removal of catheter (ROC). METHODS: A non consecutive prospective randomized study was performed in 84 patients who underwent a transurethral resection of the prostate to determine the impact of midnight versus early-morning ROC on sleep deprivation, over all discomfort to the patient. RESULTS: There was no difference in the patient experience in both groups. We found a reduced frequency during the first 6 h of ROC at midnight. However, there was an increased incidence of sleep disturbances in this group. This may in part be due to an anxiety of urge incontinence and may be allayed by appropriate counselling. There was no delay in discharge of the patients in both groups. CONCLUSION: The patients must, therefore, be given the choice of ROC either at midnight or early morning, as the advantages of a reduced frequency must be correlated with an increased incidence of sleep disturbances. PMID- 16037705 TI - Loss to follow-up in a longitudinal study on urogenital tract symptoms in Dutch older men. AB - OBJECTIVE: To describe loss to follow-up (LTFU) in a longitudinal community-based study on urogenital tract dysfunction in older men. PATIENTS AND METHODS: A cohort study of men recruited from a Dutch municipality was performed. A baseline study and two follow-up rounds--all with questionnaires and additional measurements--were performed with, on average, 2.1-year intervals. Baseline characteristics were compared between participants and non-participants in the first and in the second follow-up study. RESULTS: The response rates in the first and in the second follow-up were 78.0 and 80.0%, respectively. Various characteristics were found to be related to LTFU (i.e., more than 5% difference in response rate). Lower urinary tract symptoms were related to LTFU in the first and second follow-up. Sexual dysfunction was related to LTFU only in the second follow-up. Adjustment for confounders yielded odds ratios for the primary outcome variables (lower urinary tract symptoms, sexual dysfunction, and health status) that approximated the value of 1. LTFU according to these variables was different in men with and without other chronic illnesses. CONCLUSIONS: LTFU seems not to be related to the primary outcome variables in this study. Describing response patterns in longitudinal studies is important, especially in studies involving older participants, as often is the case in urological research. PMID- 16037706 TI - Preperitoneal prosthetic mesh hernioplasty for the simultaneous repair of inguinal hernia during prostatic surgery: experience with 172 patients. AB - OBJECTIVES: We report our experience with posterior preperitoneal prosthetic hernioplasty for inguinal hernia in patients undergoing concomitant pelvic surgery for prostatic pathologies. METHODS: 172 patients with unilateral or bilateral inguinal hernia underwent posterior preperitoneal prosthetic hernioplasty during pelvic surgery for BPH and malignant prostatic pathologies. RESULTS: There was no evidence of hernial recurrence after a mean of 30 months and no complications attributable to the hernial repairs, except for 1 patient who developed a small periprosthetic hematoma and 1 patient with a pelvic lymphocele, both spontaneously resolved. CONCLUSIONS: Posterior preperitoneal prosthetic hernioplasty for inguinal hernia combined with pelvic surgery for prostatic pathologies should be applied routinely by urologists because it is a relatively simple procedure and with a very low rate of complications and recurrence. PMID- 16037707 TI - Prospective study of estramustine phosphate for hormone refractory prostate cancer patients following androgen deprivation therapy. AB - INTRODUCTION: Estramustine phosphate (EMP) in combination with other cytotoxic agents has been widely used in clinical trials as an anti-tumor agent for the treatment of hormone-refractory prostate cancer (HRPC). However, few prospective studies have considered the efficacy of EMP monotherapy for HRPC patients following androgen-deprivation therapy (ADT), given the availability of methods to measure prostate-specific antigen (PSA) levels in the serum. We therefore initiated a prospective study to determine whether EMP is efficient for HRPC following ADT using changes in PSA levels as the major endpoint. METHODS: After a diagnosis of anti-androgen withdrawal syndrome had been excluded, 34 patients with HRPC who showed an elevated serum PSA level in 3 or more sequential tests following ADT were treated orally with 560 mg/day of EMP. The clinical stage and the median PSA value for inclusion in the study were D2 and 25.9 (range 6.5 540.8) ng/ml, respectively. Treatment was continued until evidence of disease progression reappeared or until severe adverse effects appeared. RESULTS: Of the 34 patients enrolled, 29 were evaluated, while the other 5 (15%) patients were discontinued due to severe gastrointestinal side effects. Seven of the 29 patients (24%) showed a decrease of 50% or greater in serum PSA levels from the initially elevated values, with the median duration of PSA response being 8.0 (range 2.2-18.8) months. Baseline PSA, hemoglobin, alkaline phosphatase, lactate dehydrogenase, performance status, and length of time of initial hormonal treatment did not correlate with the PSA response. With a median follow-up time of 20.0 (range 3.2-45.6) months, the cancer-specific survival rate at 2 years was 83% in the PSA responders and 44% in the non-responders. The PSA response was correlated with cancer-specific survival (p = 0.029). CONCLUSIONS: Following ADT one quarter of HRPC patients responded to EMP, with more than 50% of patients showing a decrease in PSA levels and an enhanced survival rate. PMID- 16037708 TI - Effect of acute alcohol intake on prostate tissue and serum PSA-like protein levels in rats. AB - OBJECTIVE: To investigate both the possible changes induced by acute alcohol intake on prostate tissue at the ultrastructural level and its effect on serum prostate-specific-antigen (PSA)-like protein levels in rats. MATERIALS AND METHODS: 44 male Wistar rats were included in the study in four separate groups; 7 treatment and 4 control animals were selected for each group. The treatment group received 40% ethanol (6 g/kg) while the control group was injected with the same dose of intraperitoneal isotonic saline. The first group was sacrificed after 3 h, the second group after 24 h, the third group after 72 h and the fourth group on day 7. Samples were examined by light and Jeol-100 electron microscope. Total serum PSA-like protein levels were determined by a Tosoh immunoenzymometric analyzer AIA 600. RESULTS: In groups 1-3, electron microscopy showed dilatations in the endoplasmic reticulum cisternae of prostatic acinar cells, disarrangements in apparent Golgi complex and apertures belonging to the basal labyrinth. It was also observed that there was a mass of debris cells inside the acinar lumen, and the secretory epithelium was detached from the basal membrane in some places. No pathology was found in group 4 by light and electron microscopy. In group 1, significant levels of increased serum total PSA-like protein were found compared to controls and other treatment groups (p = 0.006). CONCLUSIONS: The fundamental effect of acute alcohol intake on prostate tissue was observed in intermediate stages participating in the exocrine secretion process in the cellular organelles. At the same time, this influences the levels of serum PSA-like protein. PMID- 16037709 TI - Plasma chromogranin A in patients with prostate cancer improves the diagnostic efficacy of free/total prostate-specific antigen determination. AB - INTRODUCTION: We ascertained whether plasma chromogranin A enhances the power of serology assessing prostate cancer (PC). MATERIALS AND METHODS: We studied 56 PC and 83 benign prostatic hyperplasia (BPH) patients. In the sera we measured total prostate-specific antigen (tPSA) and free PSA (fPSA) and calculated the ratio between fPSA and tPSA (f/tPSA). In plasma samples the levels of chromogranin A (CgA) were also assayed. RESULTS: PC patients had higher CgA (p < 0.005) and tPSA (p < 0.05) levels, and a lower f/tPSA ratio (p < 0.001), than BPH patients. When f/tPSA and CgA were combined, the diagnostic sensitivity was enhanced (57-73%), while the specificity had only an 8% reduction (from 89 to 80%). CgA was only correlated to the Gleason PC score (p < 0.05). CONCLUSIONS: CgA determination in PC may enhance the diagnostic accuracy of the f/tPSA assay and provides useful information on the tumor grade. PMID- 16037710 TI - Penile sensitivity and sexual satisfaction after circumcision: are we informing men correctly? AB - OBJECTIVES: Currently no consensus exists about the role of the foreskin or the effect circumcision has on penile sensitivity and overall sexual satisfaction. Our study assesses the effect of circumcision on sexually active men and the relative impact this may have on informed consent prior to surgery. MATERIALS AND METHODS: One hundred and fifty men between the ages of 18 and 60 years were identified as being circumcised for benign disease between 1999 and 2002. Patients with erectile dysfunction were excluded from the study. The data was assessed using the abridged, 5-item version of the International Index of Erectile Function (IIEF-5). Questions were also asked about libido, penile sensitivity, premature ejaculation, pain during intercourse and appearance before and after circumcision. IIEF-5 data was analysed using two-tailed paired t test to compare pre-operative and post-operative score changes across the study group. For the rest of the questions, data was analysed using 'Sign Test', calculating two-sided p values and 95% confidence intervals. RESULTS: Fifty-nine percent of patients (88/150) responded. The total mean IIEF-5 score was 22.41 +/- 0.94 and 21.13 +/- 3.17 before and after circumcision, respectively (p = 0.4). Seventy four percent of patients had no change in their libido levels, 69% noticed less pain during intercourse (p < 0.05), and 44% of the patients (p = 0.04) and 38% of the partners (p = 0.02) thought the penis appearance improved after circumcision. Penile sensation improved after circumcision in 38% (p = 0.01) but got worse in 18%, with the remainder having no change. Overall satisfaction was 61%. CONCLUSIONS: Penile sensitivity had variable outcomes after circumcision. The poor outcome of circumcision considered by overall satisfaction rates suggests that when we circumcise men, these outcome data should be discussed during the informed consent process. PMID- 16037711 TI - Electromyographic changes after deep dorsal vein arterialization. AB - PURPOSE: To investigate the cavernosal electromyographic changes after deep dorsal vein arterialization. PATIENTS AND METHODS: Corpus cavernosum electromyographs (CC-EMG) were done in 18 patients with patent anastomosis 1 year after the operation by using coaxial needle electrodes during the flaccid state and following intracavernous 60 mg papaverine injection. RESULTS: The mean preoperative amplitude and relaxation degree measurements were found to be 77 +/- 11 microV and 34 +/- 8%, respectively. The same recordings 1 year after the operation were 140 +/- 24 microV and 76 +/- 11%, respectively. Comparisons of preoperative and postoperative values revealed statistically significant results (p < 0.0001). CONCLUSION: Smooth muscle restoration of cavernous tissue was proved by observing the postoperative increases in the amplitude and relaxation degree parameters of CC-EMG. PMID- 16037712 TI - Pre-ischemic administration of nitric oxide synthase inhibitors reduced germ cell apoptosis after spermatic vessel ligation in the rat testis. AB - INTRODUCTION: The Fowler-Stephens maneuver, a mode of spermatic vessel ligation, is a method of choice in the management of high testes. The aim of the present study is to investigate the effects of pre-ischemic administration of N(G)-nitro L-arginine methyl ester (L-NAME), a nitric oxide (NO) synthase (NOS) inhibitor, in preventing testicular ischemic damage and germ cell-specific apoptosis in rats. MATERIALS AND METHODS: 30 min before ligation of the spermatic vessels, L NAME was administered intraperitoneally to a group of rats and saline was given to another group of rats (controls). Biochemical assessments of NO and germ cell specific apoptosis were performed. RESULTS: Testicular NO levels in the L-NAME group showed significant decreases in the ipsilateral (p = 0.004) and contralateral (p = 0.015) testes in relation to those of the control group. The apoptotic indices were found in 2.3% of the L-NAME group and 3.1% of the control group. CONCLUSION: Pre-ischemic administrations of the NOS inhibitor, L-NAME, effectively decreased NO production and to some degree caused a reduction in germ cell apoptosis in the rat testes after spermatic vessel ligation. Further studies are mandatory to confirm our preliminary results and to address the potential introduction of NOS inhibitors into clinical practice. PMID- 16037713 TI - Effectiveness of vardenafil versus papaverine in penile Doppler ultrasonography. AB - PURPOSE: To evaluate erectile dysfunction, penile color Doppler ultrasound is currently considered the best method. But intracavernous injection is invasive and has adverse effects, such as prolonged erection and risk of priapism. In our study we want to evaluate whether vardenafil can be used instead of papaverine in penile Doppler ultrasonography. MATERIALS AND METHODS: A total of 24 patients with erectile dysfunction underwent color Doppler ultrasound before and after intracavernous injection of 60 mg papaverine with genital and audiovisual sexual stimulation. Peak flow and end diastolic velocity were measured in the recorded waveforms obtained 0, 1, 5, 10 and 20 min after injection. The patients also underwent color Doppler ultrasound after a 10-mg oral dose of vardenafil with genital and audiovisual sexual stimulation and at least 5 days after the prior examination. The same parameters were measured at 30, 45, 60, 75 and 90 min after the drug administration. We compared the results with the values obtained after papaverine injection. RESULTS: After oral vardenafil mean peak flow velocity significantly increased starting at 30 min and achieving a maximum value at 60 min. There were no significant differences in the 2 methods in mean peak velocity 1, 5, 10 and 20 min after papaverine injection, and 30, 45, 60, 75 and 90 min after oral vardenafil administration. Although papaverine injection is the gold standard for penile color Doppler ultrasound, it has severe adverse effects such as prolonged erection which we observed in 3 patients (12.5%) and required pharmacological detumescence. After vardenafil no severe adverse effects were observed. CONCLUSIONS: Vardenafil administration achieved increased peak flow velocity comparable to that after intracavernous papaverine injection. With no prolonged erection vardenafil is a safer alternative compared to more invasive intracavernous injection and is also an alternative for patients who fear injections. PMID- 16037714 TI - Fas ligand and TNF-related apoptosis-inducing ligand induction on infiltrating lymphocytes in bladder carcinoma by bacillus Calmette-Guerin treatment. AB - AIM: To determine the molecular mechanisms underlying the efficacy of bacillus Calmette-Guerin (BCG) therapy against superficial carcinoma of the urinary bladder, we evaluated the expression of cytotoxic molecules on tumor-infiltrating lymphocytes before and after therapy. METHODS: Immunofluorescence staining allowed the specific detection of Fas, Fas ligand (FasL), and tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) expression on tumor cells and the respective leukocyte populations in biopsy samples from 6 patients. RESULTS: Significant increases in the infiltration of FasL- and TRAIL-expressing CD4+ T cells and macrophages and FasL-expressing CD8+ T and NK cells were observed after BCG instillation in bladder carcinoma. Moreover, Fas expression was upregulated on tumor cells after BCG instillation. CONCLUSION: The data suggested that the enhanced infiltration of FasL- and/or TRAIL-expressing leukocytes (CD4+ T cells, CD8+ T cells, natural killer cells and macrophages) and the induction of Fas expression on tumor cells may play an important role in the therapeutic effect of BCG instillation. PMID- 16037715 TI - Spontaneous external drainage of renal abscess through persistent nephrostomy tract eleven years after percutaneous nephrolithotomy. AB - A unique case of spontaneous external drainage of a renal abscess through a persistent nephrostomy tract is reported in a patient with a history of two subsequent percutaneous nephrolithotomies 11 years earlier. Nephrostomy tracts can open up even after several years to provide an exit path for infected urine or renal abscess drainage. PMID- 16037716 TI - Adult granulosa cell tumour of the testis and bony metastasis. A report of the first case of granulosa cell tumour of the testicle metastasising to bone. AB - Granulosa cell tumour is among the rarest forms of testicular cancer. Only 21 cases have been reported to date and only 4 have metastasised. All documented metastases are intra-abdominal. We present the first case of granulosa cell tumour of the testis metastasising to bone (tibia), presenting 6 years after orchidectomy. This highlights the need to maintain a high index of suspicion for extra-abdominal metastases during long-term follow-up in patients with previous testicular granulosa cell tumour. PMID- 16037717 TI - Testicular granulocytic sarcoma without systemic leukemia. AB - This case report describes a unilateral testicular granulocytic sarcoma or chloroma. Because of the relatively immature nature of the tumor cells, the histological diagnosis can be difficult. Granulocytic sarcomas are well known in patients with systemic leukemia and can sometimes precede a systemic leukemic outcome. A solitary granulocytic sarcoma not followed by a hematological proliferation of the myelocytic stem cells is very rare. No prognostic factors that are able to predict a systemic outcome are known. Therefore, in this case with no signs of systemic disease, we adopted a wait-and-see policy after radical orchidectomy. Up to now, after a follow-up period of 7 years, the patient is still free of disease. Diagnosis and therapy of this urologic disease are discussed and the literature is reviewed. PMID- 16037718 TI - Perception of nurse experts on the contribution of nursing interventions to NOC nursing outcomes in general hospitals in Korea. AB - The purpose of this study was to identify the perception of nurse experts on the contribution of nursing interventions to Nursing Outcomes Classification NOC nursing outcomes. A nursing outcome is a nursing-sensitive patient outcome primarily affected by nursing interventions. As one of the standardized language systems of nursing outcomes, the NOC must be examined for applicability before it is used in Korea. Data were collected in February and March 2003 using a 5-point Likert scale. For data collection, 230 quality improvement (QI) or quality assurance (QA) nurses from general hospitals in Korea were asked to rate the extent that nursing interventions contribute to each of the NOC nursing outcomes (2,000) in their hospitals. Ninety-six nurses from 63 hospitals responded and the response rate was 41.7%. Mean scores for perception of contribution of nursing interventions to each of the NOC nursing outcomes ranged from 2.18 to 4.54. Vital Signs Status had the highest score (M=4.54), and Abuse Recovery: Financial, the lowest score (M=2.18). Of the seven NOC domains, the mean score was highest for Physiologic Health (M=3.91) and lowest for Community Health (M=2.92). Of the 29 NOC classes, the mean score for perceived contribution was highest for Metabolic Regulation (M=4.32) and lowest for Community Well-Being (M=2.92). Participants perceived that nursing interventions in general hospitals in Korea contributed, at least to a certain extent, to most of the NOC nursing outcomes. Based on these results, NOC should have relatively good applicability in Korea. PMID- 16037719 TI - Barriers and attitudes to research among nurses in one hospital in Korea. AB - PURPOSE: Nursing research is recognized as an essential basis for the continuous development of the scientific nursing knowledge and practice. An understanding of the barriers and attitudes to research among nurses is important to improve clinical nursing research activities. METHOD: Data were collected from nurses at a general hospital, located in D city, using a self-reported questionnaire, from March 15th to April 4th, 2003, and 133 questionnaires were used for the analyses. A 5 point scale instrument was used to measure barriers and attitudes towards nursing research, with a Cronbach's alpha of .85 and .89, respectively. RESULTS: Nurses had limited experience in research and were not well prepared to conduct it. Lack of experience, training and time were the major barriers for conducting research. However, the nurses had positive attitudes toward research. There was a difference in barriers (t=2.68, p=.02) and research attitudes (t=-2.74, p=.00) according to the nurses' position. Also, the nurses' research experience influenced research attitudes (t=-3.27, p=.00). The degree of research preparation (F=6.98, p=.00) had an effect on the attitudes toward nursing research but not on the barriers. CONCLUSION: There is an urgent need to promote both the ability and environment for clinical nurses to actively participate in research. As the majority of nurses had little experience and low confidence in research and perceived many barriers, this promotion can be accomplished by enhancing their confidence in research through continuing education and by gradually eliminating the barriers to research. PMID- 16037720 TI - A study of the relationships between daily life stress, self-efficacy and university student life adjustment. AB - PURPOSE: This correlation study was undertaken to examine the relationships between daily life stress, self-efficacy and university student life adjustment and to analyze the self-efficacy affecting that university student life adjustment. METHODS: 265 university students were recruited from several participating required undergraduate classes. They were surveyed as the subject of this study. Data collection was conducted through the use of questionnaires. RESULTS: The university life adjustment level showed a maximum score of 9 with the mean score 5.22. A negative correlation was found between daily life stress and university life adjustment. Also, there was positively correlation between self-efficacy and university life adjustment. Self-efficacy and daily life stress accounted for 23% of the variance in university student life adjustment. CONCLUSION: According to the results, self-efficacy is a useful concept in helping overall university life adjustment. Therefore, university student consultation office or nurse should consider the program based on self efficacy in order to help university students to better adapt to university life. PMID- 16037721 TI - The effect of job stress and lifestyle on blood lipid levels in male aircrew personnel. AB - INTRODUCTION: [corrected] Cardiovascular disease has the potential to lead to sudden in-flight incapacitation and permanent grounding. The aims of this study are to examine the relationships between lifestyle, job stress and blood lipid levels of male aircrew personnel of a Korean airline and to identify which factors influence their hyperlipidemia. METHODS: Two hundred sixteen male aircrew personnel completed a questionnaire by self-report and consented to participate in the study. The questionnaire collected data related to job stress, life style, serum cholesterol levels and general characteristics of the aircrew. The cholesterol levels of the subjects were collected from their most recent health check-up records. Subjects were divided into two groups (the desirable group and the risk group) based on their serum cholesterol level, 200 mg/dl. RESULTS: Mean age and marital status were significantly different between the two groups. More subjects in the risk group had habits of eating high lipid foods, while more subjects in the desirable group exercised more frequently than the risk group. In logistic regression analysis, after controlling age and marital status, types of working situation (domestic duty or international duty, odds ratio=.390, p=.018), diet (odds ratio=.429, p=.037), and exercise (odds ratio=.320, p=.055) were influencing factors on aircrew's serum cholesterol levels. CONCLUSIONS: The cholesterol level of aircrew personnel is closely related to their lifestyle, such as lipid diet and exercise. The type of work situations, e.g. staying in an airplane for long periods of time or staying abroad, may influence these diet patterns and exercise habits. PMID- 16037722 TI - Study on nursing college students' subjectivity in their attitude toward jobs. AB - PURPOSE: The purpose of the study was to categorize nursing students' subjectivity in their attitude toward their jobs, and thereby understand the differences among these attitude types. METHODS: The study used a Q-method to measure nursing students' attitude toward jobs identity types. In-depth and objective interviews and literature review formed Q sample. The P sample consisted of 25 nursing students. RESULTS: The results of the study show that nursing students can be categorized into three types, depending on their attitudes toward their jobs. The first type, "interest-oriented" students, strongly disagree to the following: giving priority to job over marriage, standing unfair treatment in the workplace, the importance of promotion opportunity, irresponsibility, and uncertainty. The "reward-oriented" students, on the other hand, strongly disagree to the following: indifference to career prospects, employment-related relocation of residence, irresponsibility, standing difficulties, and compromises with others. The third type of nursing students is the possession-oriented students, who strongly disapprove of irresponsibility, refusal to compromise with reality, standing unfair job allocation or promotion and career uncertainty. CONCLUSIONS: The study on nursing college students' attitude toward their jobs is meaningful in the following aspects: First, the study clarifies nursing college students' attitudes toward their job by categorizing it. Second, the study confirms the changing attitudes of nursing students toward jobs with the change of times and calls for proper educational programs to foster healthy career attitudes. Third, proper decision-making as regards jobs and job allocation for nurses, or their career attitudes, is beneficial to individuals, the medical industry, and society. PMID- 16037723 TI - Effect of asthma management education program on stress and compliance of patients with allergic asthma to house dust mite. AB - PURPOSE: This study was designed to examine the effect of asthma management education program applied to allergic asthma patients receiving immunotherapy due to house dust mite on their stress and compliance with health care regimens. METHODS: A quasi experimental design with non-equivalent control group and non synchronized design was used. The subjects of this study were 61 patients who were receiving immunotherapy at intervals of a week after their symptoms were diagnosed as house dust mite allergic asthma at the pulmonary department of a university hospital in Seoul. They were divided into an experimental group of 29 patients who received asthma management education and a control group of 32 patients. The asthma management education program was composed of group education (once) and reinforcement education (three times) with environmental therapy and immunotherapy to house dust mite. RESULTS: Stress significantly decreased in the experimental group compared to that in the control group. Compliance with health care regimens significantly increased in the experimental group compared to that in the control group. CONCLUSIONS: The results suggested that the asthma management education program is effective for the management of stress and the improvement of compliance in patients with allergic asthma to house dust mite. PMID- 16037724 TI - Peer and parental influences on adolescent smoking. AB - PURPOSE: The purpose of this study was to determine the relationship between peer and parental factors and smoking behavior of adolescents in urban cities and to investigate whether there are gender differences. METHODS: A stratified and random cluster sampling design was used to obtain a cross-sectional sample of high school students in two urban cities. The sample consisted of 512 Korean adolescents (256 boys and 256 girls) aged 15 to 18 (mean age 16.7+/-.58). Self reported questionnaire consisted of adolescent smoking behavior, peer smoking and alcohol use, parental smoking and alcohol use, father-mother-peer relationships and perceived social support from peers and parents. Multiple logistic regression analysis was used to examine the hypothesized model. RESULTS: The findings showed that peer and parental factors accounted for 30.3% of the variance in adolescent smoking and peer smoking was most strongly associated with adolescent smoking behavior (OR=10.18). In addition, peer smoking (OR=4.71), peer alcohol use (OR=4.21), and peer relationships (OR=1.03) were significantly associated with boys' smoking behavior. In girls, peer smoking (OR=26.50) and parent smoking (OR=5.48) were significantly associated with smoking behavior. CONCLUSIONS: Consistent with previous findings, peer smoking is a significant factor on adolescent smoking. Specifically, boys would be more influenced from peers than girls. Therefore, smoking prevention programs for adolescents might be focused on the social context such as, resisting to peer pressure and enhancing the self efficacy to control. PMID- 16037725 TI - Fatigue and its related factors in Korean patients on hemodialysis. AB - PURPOSE: This study examined the characteristics of fatigue and the relationship between fatigue and its related factors in Korean patients on hemodialysis. METHODS: A cross-sectional correlational study was conducted with 104 patients on hemodialysis in Seoul, Korea. FINDING: Of a total of 104 subjects, eighty-one (77.9%) complained of fatigue. Fatigue severity was measured by the self-rating Visual Analogue Scale-Fatigue (VAS-F) with a mean score of 36.5 (SD=17.49, range 2--81). The mean duration of fatigue was 3.8 hours (SD=5.3, range 0--24). Depression was most significantly correlated with fatigue (beta=.43, p<.00), with inter-dialytic weight gain (beta=.25, p<.05) being the second most significant correlate. CONCLUSION: This study shows that nursing interventions for patients who experience fatigue while on hemodialysis should be focused on both psychological problems, such as depression, as well as on physiological problems, such as inter-dialytic weight gain. PMID- 16037726 TI - Spirituality: concept analysis based on hybrid model. AB - PURPOSE: The purpose of this study was to obtain a clearer understanding of spirituality and examine the process of spirituality through defining the meaning and attributes of spirituality. METHOD: Concept analysis was done in the three phases, theoretical phase, fieldwork phase, and analytical phase suggested in the Hybrid Model. Five people participated in the fieldwork phase. RESULTS: Spirituality is activated through self-awareness which occurs as spirit being activated through self-introspection, and through restoration of the relationship with Supreme Being. This interconnectedness with Supreme Being has an absolute impact on one's harmonious interconnectedness with self and neighbors, thus leads all the critical attributes of spirituality to be revealed. The core energy of this harmonious interconnectedness is love. When activated, it has a great impact on an individual as integrative energy, leads one to go beyond everyday experience as well as to have new perspectives, and to live a satisfactory life in every aspect. CONCLUSION: The results of this study suggest that promotion of connectedness is the most important element in spiritual nursing interventions. The results can also be used effectively in developing spirituality assessment scales and theory. PMID- 16037727 TI - The factors influencing the compliance of breast self-examination of middle-aged women. AB - PURPOSE: This cross-sectional survey was conducted to described the compliance of Breast Self-Examination of middle-aged women using a convenient sample, and to examine relationships between the compliance of BSE and Health Beliefs, and the influencing factors on the compliance of BSE. METHODS: The subjects were 373 literate volunteers who were from 41 to 60 years of age who visited 6 public health centers. From June 7, 2004 to August 20, 2004, data were collected by 5 research assistants using a self-report questionnaire. The questionnaire was used to obtain information on the general characteristics, knowledge, health beliefs, and compliance of BSE. RESULTS: The findings of this study suggested that there were significant differences in the scores of the perceived susceptibility and severity between compliers and non-compliers of the BSE. BSE compliance was significantly correlated with knowledge, perceived susceptibility, and perceived severity. The most powerful predictor of BSE compliance was the perceived susceptibility. The perceived susceptibility, the perceived severity, the knowledge and educational level accounted for 41.8% of the variance in middle aged women's BSE compliance. CONCLUSION: Increase in knowledge about breast cancer, with a concomitant increase in both perceived susceptibility and perceived severity could produce a subtle cue or motivating force sufficient to affect a behavior change. Further research is needed to examine the qualitative difference between BSE and other early detection behaviors. PMID- 16037728 TI - Relationships of mood disturbance, symptom experience, and attentional function in women with breast cancer based upon the theory of unpleasant symptoms. AB - PURPOSE: The purpose of this study was to identify direct, mediating, and moderating relationships of mood disturbance, symptom experience, and attentional function in Korean women with breast cancer based upon a middle-range theory of unpleasant symptoms. METHODS: This study used a cross-sectional, correlational design. A convenience sample of 125 women receiving chemotherapy for breast cancer was recruited from a university hospital in South Korea. The women completed questionnaires on mood disturbance, symptom experience, and attentional function using the Linear Analogue Self-Assessment Scale, the Symptom Experience Scale, and the Attentional Function Index, respectively. RESULTS: Each mood disturbance and symptom experience showed a significant relationship with attentional function. Symptom experience did not act as a mediator between mood disturbance and attentional function, but it did act as a moderator: patients with a higher level of mood disturbance exhibited a lower level of attentional function when their symptoms were at the level of medium, but not when their symptoms were either high or low. CONCLUSION: This suggests that clinical interventions for attenuating the influence of mood disturbance on attentional function may be effective only in women experiencing medium level of symptoms. PMID- 16037729 TI - Modeling adherence to therapeutic regimens in patients with hypertension. AB - PURPOSE: This study was done to identify and test a model of the psychosocial variables that influence adherence to therapeutic regimens in patients with hypertension. METHOD: A convenience sample of 219 patients with hypertension who were enrolled in an outpatient clinic of a cardiovascular center in Korea participated in the study. They completed self-administered questionnaires anonymously. The questionnaire was based on the Social Action Theory model and a literature review. The explanatory model was constructed and tested using structural equation modeling in order to examine the effects within the model. RESULTS: The results of this study showed that perceived self-efficacy was the strongest factor influencing patient adherence in this sample. Adherence to therapeutic regimens in patients with hypertension was influenced by self efficacy, patient-provider relationship, social support, and depression. CONCLUSIONS: Adherence to therapeutic regimens in patients with hypertension was most strongly influenced by self-efficacy. These findings suggest that nursing interventions to promote patient adherence should focus on the promotion of self efficacy including improvement in patient-provider relationship and social support, and reduction in depression. PMID- 16037730 TI - Development and analysis of the effects of caregiver training program on aggressive behavior in elders with cognitive impairment. AB - PURPOSE: The purposes of this study were to 1) describe the type and frequency of aggressive behavior of cognitively impaired nursing home resident, 2) develop a caregiver training program on prevention and management of aggressive behavior, 3) examine the effects of caregiver training program on the incidence of aggressive behavior of cognitively impaired nursing home resident, and 4) examine the effects of caregiver training program on nursing staff's aggressive behavior management skills. METHODS: One-group, time series, quasi-experimental design with a pre-test and two post-tests was used. Data were collected from cognitively impaired home residents (N=32) and nursing staff (N=36) in a proprietary nursing home using Ryden Aggression Scale I, II, and Aggressive Behavior Management Scale. Data were entered and analyzed by descriptive statistics and repeated measures ANOVA. RESULTS: Incidence of aggressive behavior was high with a mean score of 3.09 (SD=3.11) at baseline. Caregiver training program was developed based on Progressively Lowered Stress Threshold (PLST) model and gerontological and psychiatric literature. The mean scores of aggressive behavior at baseline, Post I, and II did not differ significantly although the difference approached to the significant level (F=2.925, p=.066). Nursing staff's aggressive behavior management skills increased at Post I, and at Post II when compared to baseline, and the difference was significant (F=12.736, p=<.001). CONCLUSION: Caregiver training program showed potential impact on reduction of aggressive behavior in elders with cognitive impairment and was effective in increasing nursing staff's aggressive behavior management skills. PMID- 16037731 TI - Effect of preemptive gabapentin on postoperative pain relief and morphine consumption following lumbar laminectomy and discectomy: a randomized, double blinded, placebo-controlled study. AB - Synergism between gabapentin and morphine in treating incisional pain has been demonstrated in animal experiments and clinical studies. The efficacy of gabapentin for treatment of perioperative pain remains controversial. This study was designed to detect the influence of gabapentin premedication on morphine consumption in the immediate postoperative period in patients undergoing lumbar laminectomy and discectomy. Either gabapentin 800 mg (in two equally divided doses) or placebo was given preoperatively to 60 adult patients undergoing elective lumbar laminectomy or discectomy in a double-blinded, placebo controlled, randomized study. Standard general anesthesia was given to all the patients. Morphine was administered via patient-controlled analgesia pump in the immediate postoperative period for first 8 hours. Pain at rest and on movement was assessed using a Verbal Rating Scale (VRS) every 2 hours for the first 8 postoperative hours. There were no differences in demographics or surgical duration between the two groups. The amount of fentanyl administered in the intraoperative period was similar between the two groups. In the postoperative period, the VRS score for pain at 0, 2, 4, 6, and 8 hours was not significantly different between the two groups. Highest median VRS score was recorded at 0 hours postoperatively in both groups (VRS: rest = 6, movement = 8 in placebo group; rest = 6, movement = 8 in gabapentin group). Total morphine consumption and side effects were similar in the two groups. Gabapentin does not decrease the morphine requirement or morphine side effects in the immediate postoperative period following lumbar laminectomy and discectomy. PMID- 16037732 TI - Comparison of low-dose intrathecal and epidural morphine and bupivacaine infiltration for postoperative pain control after surgery for lumbar disc disease. AB - This prospective, blinded, placebo-controlled study was performed to compare the postoperative analgesic efficacy of low-dose intrathecal and epidural morphine with paraspinal muscle infiltration of bupivacaine in lumbar discectomy cases. Eighty ASA I-III adult patients undergoing elective surgery for lumbar disc disease were enrolled in the study. Patients were randomized to four groups by envelopes. Study groups were as follows: group 1 (n = 20), intrathecal morphine 0.1 mg; group 2 (n = 20), epidural morphine 2 mg; group 3 (n = 20), 30 mL of bupivacaine 0.25% paraspinal muscle infiltration; group 4 (n = 20), 30 mL of saline paraspinal muscle infiltration before wound closure. Recorded parameters were time to response to painful and verbal stimuli and postoperative pain assessed at 30 minutes and 2, 4, 6, 8, 12, and 24 hours by Visual Analog Scale (VAS) and Numeric Pain Scale (NPS). Hemodynamic data, sedation scores, and side effects were also recorded. Meperidine and naproxen sodium were used for postoperative analgesia. Follow-up was performed by a blinded investigator. Mean VAS scores were lower in groups 1 and 2 at 30 minutes (P < 0.05). Mean VAS score of group 2 was lower than that of group 4 at 4 hours postoperatively (P < 0.05). Mean NPS scores were lower in groups 1 and 2 at 2, 4, and 6 hours (P < 0.05) and in group 2 at 8 hours compared with the other groups. The number of patients requiring meperidine at early postoperative phase (0-6 hours) was less in groups 1 and 2 compared with groups 3 and 4 (P < 0.05). There were no statistically significant differences in the late postoperative analgesic requirements, after correction for multiple testing. In conclusion, low-dose intrathecal and epidural morphine provide lower postoperative pain scores and a reduction in early postoperative analgesic requirement with insignificant side effects compared with paraspinal bupivacaine or saline infiltration. PMID- 16037733 TI - No difference in emergence time and early cognitive function between sevoflurane fentanyl and propofol-remifentanil in patients undergoing craniotomy for supratentorial intracranial surgery. AB - Balanced anesthesia with sevoflurane-fentanyl has been widely accepted as anesthetic management for neurosurgery. Propofol-remifentanil regimen has been successfully used in various surgical settings, but a comprehensive comparison of sevoflurane-fentanyl and propofol-remifentanil anesthesia in patients undergoing craniotomy for supratentorial intracranial surgery has not yet been done. The aim of this prospective, randomized, open-label clinical trial was to compare clinical properties of sevoflurane-fentanyl with propofol-remifentanil anesthesia in patients undergoing supratentorial intracranial surgery. The primary endpoint was to compare early postoperative recovery and cognitive functions within the two groups; we also evaluated hemodynamic events, vomiting, shivering, and pain. One hundred twenty patients (64 males; age 15-75 years) were randomized to either total intravenous anesthesia (group T) or sevoflurane anesthesia (group S). Emergence and extubation times and cognitive function (Short Orientation Memory Concentration Test [SOMCT]) were compared in the two groups. Brain swelling, incidence of hypotensive and hypertensive episodes, postoperative vomiting, shivering, and pain were also analyzed. The mean emergence time (12.2 +/- 4.9 minutes for group S versus 12.3 +/- 6.1 minutes for group T; P = 0.92) and extubation time (18.2 +/- 2.3 minutes for group S versus 18.3 +/- 2.1 minutes for group T; P = 0.80) were similar in the two groups. Average SOMCT scores, both 15 minutes after extubation (25.6 +/- 4.9 in group S versus 23.9 +/- 7.5 in group T; P = 0.14) and 45 minutes after extubation (27.3 +/- 2.2 in group S versus 26.0 +/ 5.1 in group T; P = 0.07) were also comparable. Brain swelling was present in seven and five patients in groups S and T, respectively (P = 0.76). Hypotension was present in 12% (group S) and 28% (group T) of patients (P = 0.02). Hypertension was present in 17% of patients in group S and 40% of patients in group T (P = 0.0046). Shivering was present in 18% and 25% of patients in groups T and S (P = 0.37). Our study demonstrates that there is no patient benefit of using total intravenous anesthesia with an ultra-short-acting opioid over the conventional balanced volatile technique in terms of recovery and cognitive functions. PMID- 16037734 TI - Scalp infiltration with bupivacaine plus epinephrine or plain ropivacaine reduces postoperative pain after supratentorial craniotomy. AB - Local anesthetic infiltration has been proposed to decrease postoperative pain. The aim of this study was to determine whether scalp infiltration with bupivacaine or ropivacaine would improve analgesia after supratentorial craniotomy for tumor resection. Eighty patients were recruited into a randomized double-blind study. Infiltration was performed after skin closure with 20 mL of saline 0.9% (placebo group, n = 40), of 0.375% bupivacaine with epinephrine 1:200,000 (bupivacaine group, n = 20), or of 0.75% ropivacaine (ropivacaine group, n = 20). Postoperative analgesia was provided with patient-controlled morphine IV analgesia (PCA). The study was continued until PACU discharge, which occurred early in the morning following surgery. Results are reported on 37 patients in the placebo group, 20 in the bupivacaine group, and 19 in the ropivacaine group because 4 patients experienced postoperative complications and were excluded from the study. Morphine titration at arrival in the postanesthesia care unit was necessary more often in the placebo group (62% of the patients) than in the 2 treated groups (19% in each, P = 0.02). The median quantity of morphine administered during the first 2 postoperative hours, including initial titration administered by a nurse and PCA-administered morphine, was lower in each treated group than in the placebo group (P < 0.01). The median morphine consumption up to the 16th postoperative hour was not significantly different among the 3 groups. There was no difference in the visual analogue scale scores among the 3 groups at any time. Scalp infiltration with either bupivacaine or ropivacaine had a statistically significant effect on morphine consumption during the first 2 postoperative hours. PMID- 16037735 TI - Protective effect of propofol against kainic acid-induced lipid peroxidation in mouse brain homogenates: comparison with trolox and melatonin. AB - This study compared the effectiveness of propofol with that of trolox and melatonin for reduction of lipid peroxidation in vitro. Lipid peroxidation was induced by addition of kainic acid (KA; 10 mM), hydrogen peroxide (H2O2; 10 mM), or ferrous ammonium sulfate (5 microM) to mouse brain homogenate, and thiobarbituric acid-reactive substances (TBA-RS) were used as a marker of lipid peroxidation. Propofol, trolox, and melatonin reduced KA-, H2O2-, and ferrous ammonium sulfate-induced lipid peroxidation in a concentration-dependent manner. In reducing KA-induced lipid peroxidation, 50% inhibitory concentration (IC50) values of antioxidants were as follows: propofol (11.33 mM), trolox (4.00 mM), and melatonin (9.72 mM). In reducing H2O2-induced lipid peroxidation, IC50 values of antioxidants were as follows: propofol (56.86 mM), trolox (33.34 mM), and melatonin (26.63 mM). In reducing ferrous ion-induced lipid peroxidation, IC50 values of antioxidants were as follows: propofol (49.57 mM), trolox (60.35 mM), and melatonin (22.02 mM). Under the in vitro conditions of this experiment, propofol was an excellent and a very potent antioxidant in inhibiting KA-, H2O2-, and ferrous ion-induced lipid peroxidation in mouse brain homogenates. We conclude that the antioxidant properties of propofol at clinically relevant anesthetic concentrations may have a neuroprotective effect. PMID- 16037736 TI - The effect of phenytoin on rocuronium-induced neuromuscular block in the rat phrenic nerve-hemidiaphragm preparation. AB - Anticonvulsant therapy alters the action of nondepolarizing muscle relaxants. We determined the effects of acute and chronic administration of phenytoin on rocuronium-induced neuromuscular block using the rat phrenic nerve-hemidiaphragm preparation. Rats were divided into 3 groups: a saline control group (n = 10), an acute phenytoin-treated group (n = 30), and a chronic phenytoin-pretreated group (n = 30). Phrenic nerve-hemidiaphragm was dissected, mounted in a bath containing oxygenated Krebs solution, and the nerve was stimulated at supramaximal intensity. Single twitch responses were recorded by physiogram. In the acute phenytoin-treated group, acute effects of phenytoin were determined based on the phenytoin concentration of 1, 10, or 100 microg/mL in the bath. The chronic effects of phenytoin were determined using phrenic nerve-diaphragms from rats pretreated with phenytoin (50 mg/kg/d) for 1, 7, or 28 days. In rats with phenytoin 100 microg/mL in the bath, all concentrations of rocuronium produced twitch depression significantly different from those of other groups (P < 0.05), and the concentration-response curve shifted to the left. In rats with phenytoin 10 microg/mL in the bath, the effective concentrations for 50%, 90%, and 95% twitch depression values were significantly different from those of the control group (P < 0.05). In chronically (28 days) phenytoin-pretreated rats, the concentration-response curve significantly shifted to the right (P < 0.05). These findings show that acute administration of phenytoin augmented the neuromuscular blocking effects of rocuronium, whereas chronic phenytoin treatment causes resistance to the neuromuscular blocking effects of rocuronium in target organs. PMID- 16037737 TI - Plasma ropivacaine levels during awake intracranial surgery. AB - Plasma ropivacaine levels produced by routine scalp infiltration for field block were assayed in eight patients undergoing awake intracranial surgery. Levels peaked within approximately 13 minutes, indicating rapid systemic absorption of ropivacaine. Potentially toxic plasma drug levels were achieved in three of eight patients, although no untoward effects attributable to elevated levels of ropivacaine were observed. Clinical vigilance is indicated during scalp infiltration with ropivacaine. PMID- 16037739 TI - Does preemptive use of gabapentin have no effect on postoperative pain and morphine consumption following lumbar laminectomy and discectomy? PMID- 16037740 TI - Response to: Ten years of experience with paediatric neuroendoscopic third ventriculostomy features and perioperative complications of 210 cases. PMID- 16037741 TI - Anesthesia for emergency craniotomy in a patient with von Hippel Lindau disease with pheochromocytoma. PMID- 16037743 TI - Difficult intubation related to ankylosis of temporomandibular joint after craniotomy. PMID- 16037744 TI - Exceptional clinical observation: total brain ischemia during normal intracranial pressure readings caused by obstruction of the outflow of a neuroendoscope. PMID- 16037745 TI - Postoperative hallucinations following propofol infusion in a neurosurgical patient: a diagnostic dilemma. PMID- 16037746 TI - JCAHO's most challenging standards. PMID- 16037747 TI - Postmenopausal hormone therapy and cardiovascular disease: one view of the elephant. PMID- 16037748 TI - Determining the effects of androgen therapy on sexual well-being: a complex challenge. PMID- 16037749 TI - The social and cultural context of sexual function among middle-aged women. PMID- 16037750 TI - Compliance with hormone therapy after Women's Health Initiative: who is to blame? PMID- 16037751 TI - Determinants of the effect of estrogen on the progression of subclinical atherosclerosis: Estrogen in the Prevention of Atherosclerosis Trial. AB - OBJECTIVE: To determine the extent to which the estrogen-induced changes in lipids and markers of carbohydrate metabolism explain the beneficial effect of estrogen therapy on the progression of carotid artery intima-media thickness (IMT) in postmenopausal women. DESIGN: A randomized, double-blind, placebo controlled, single-center trial enrolling 222 postmenopausal women 45 years and older without cardiovascular disease and with low-density lipoprotein (LDL) cholesterol levels of 3.37 mmol/L or greater (> or = 130 mg/dL). Intervention was unopposed micronized 17beta-estradiol versus placebo. Measurements were made using high-resolution B-mode ultrasonography to measure carotid artery IMT at baseline and every 6 months on-trial. RESULTS: Progression of carotid IMT was inversely related to on-trial high-density lipoprotein (HDL) cholesterol (P = 0.04) and was directly related to on-trial LDL-cholesterol (P = 0.005). Compared with placebo, women randomized to estradiol showed a higher mean on-trial HDL cholesterol level and a lower mean on-trial LDL-cholesterol level. In contrast, fasting glucose, insulin, and hemoglobin A1C were lowered and insulin sensitivity increased with estradiol therapy, but the changes were not related to carotid IMT progression. On-trial HDL-cholesterol and LDL-cholesterol were significant independent determinants of carotid IMT progression, jointly explaining 30% of the treatment effect of unopposed estrogen on the progression of carotid IMT. CONCLUSION: Unopposed 17beta-estradiol reduced carotid IMT progression in postmenopausal women in part by increasing HDL-cholesterol and decreasing LDL cholesterol. Although women randomized to estradiol showed improvement in all the markers of carbohydrate metabolism, these factors did not play a significant role in carotid IMT progression. PMID- 16037752 TI - Combined esterified estrogens and methyltestosterone versus esterified estrogens alone in the treatment of loss of sexual interest in surgically menopausal women. AB - OBJECTIVE: To compare the effect of esterified estrogens and methyltestosterone versus esterified estrogens alone on diminished sexual interest in surgically menopausal women. DESIGN: This randomized, double-blind study compared the effect of combined esterified estrogens (1.25 mg) and methyltestosterone (2.5 mg) (EE/MT) versus esterified estrogens (1.25 mg) alone (EE) for 8 weeks. Several different sexual function questionnaires were used to measure response to therapy. Changes from baseline in sexual interest/function and hormone levels were evaluated after 4 and 8 weeks of treatment. RESULTS: A total of 102 women were randomized into the study; 52 (age range, 32-61 years) to EE/MT and 50 (age range, 33-62 years) to EE. After 8 weeks, significant differences between treatments were not seen in the Changes in Sexual Functioning Questionnaire (CSFQ F-C) sexual desire/interest subscale score, the primary efficacy variable. In contrast statistically significant between-treatment differences were found for several secondary efficacy variables including Menopausal Sexual Interest Questionnaire (MSIQ) sexual interest/desire score, CSFQ-F-C arousal/erection subscale score and Women's Health Questionnaire sexual functioning subscale score. The mean serum concentration of bioavailable and free testosterone significantly increased, approximately doubling between baseline and the end of the study in patients receiving EE/MT, with a significant (P < 0.001) between treatment difference. The mean serum concentration of sex hormone-binding globulin significantly decreased to less than one third of the pretreatment levels in patients receiving EE/MT (P < 0.001). Both treatments were well tolerated. CONCLUSIONS: The mixed results seen with the different sexual function questionnaires may be due to the CSFQ-F-C's lack of specificity for this population. Increased levels of bioavailable and free testosterone paralleled the improved MSIQ item scores. Both the EE and EE/MT treatments were well tolerated. PMID- 16037753 TI - Correlates of sexual function among multi-ethnic middle-aged women: results from the Study of Women's Health Across the Nation (SWAN). AB - OBJECTIVE: To examine sexual function in a cohort of Baby Boomer women of diverse racial/ethnic backgrounds; to compare differences between pre-and early perimenopausal women; and to identify sociodemographic, health-related, and psychosocial (including psychological, behavioral, and relationship) factors related to sexual function. DESIGN: Six domains of sexual function were studied in 3,167 women in the baseline cohort of the Study of Women's Health Across the Nation (SWAN). Participants were 42 to 52 years old, pre-or early perimenopausal, and not using hormones. The study sample included non-Hispanic white, African American, Hispanic, Chinese, and Japanese women. RESULTS: Early perimenopausal women reported greater pain with intercourse than premenopausal women (P = 0.01), but the two groups did not differ in frequency of sexual intercourse, desire, arousal, or physical or emotional satisfaction. Variables having the greatest association across all outcomes were relationship factors, the perceived importance of sex, attitudes toward aging, and vaginal dryness. Despite controlling for a wide range of variables, we still found ethnic differences for arousal (P < 0.0001), pain (P = 0.03), desire (P < 0.0001), and frequency of sexual intercourse (P = 0.0003). African American women reported higher frequency of sexual intercourse than white women; Hispanic women reported lower physical pleasure and arousal. Chinese women reported more pain and less desire and arousal than the white women, as did the Japanese women, although the only significant difference was for arousal. CONCLUSIONS: Relationship variables, attitudes toward sex and aging, vaginal dryness, and cultural background have a greater impact on most aspects of sexual function than the transition to early perimenopause. PMID- 16037754 TI - Hormone use and patient concerns after the findings of the Women's Health Initiative. AB - OBJECTIVE: To assess behaviors and concerns related to hormone therapy after the findings of the Women's Health Initiative (WHI). DESIGN: A survey was mailed to a random sample of 1,200 women identified through the pharmacy database as taking one of two estrogen + progestogen therapies (EPT) during the 6-month period before the publication of WHI findings. Questions included hormone use history, changes in usage, an assessment of symptoms, symptom changes, health behavior changes, use of alternative therapies, and demographics. RESULTS: The response rate was 70%, with women in their 60s and those receiving hormone therapy for 5 or more years were more likely to respond (P < 0.05). The majority had started hormones for symptom relief (69%) and expected to continue use. Many reported discontinuation (63%) or modifying their medication (18%). Half of these women stopped then restarted, the other half changed products. Women in their 50s were more likely to remain on hormones than older women (P < 0.01), and those taking ethinyl estradiol and norethindrone acetate were more likely to remain on their medication than those on conjugated estrogens (43% vs 29%, P < 0.01). Little change was reported in exercise and 19% increased their calcium intake. Patient concerns fell into five major categories: long-term effects, symptom control, breast cancer risk, bone health, and cognitive function. CONCLUSIONS: Women seem to be heeding the warnings about hormones but remain concerned about the potential long-term sequelae and symptom control. More research is needed to identify safer approaches to symptom relief and to address the concerns expressed. PMID- 16037755 TI - The impact of the Women's Health Initiative Randomized Controlled Trial 2002 on perceived risk communication and use of postmenopausal hormone therapy in Germany. AB - OBJECTIVE: The purpose of this representative, nationwide telephone survey was to collect information about postmenopausal hormone therapy (HT) use in relation to women's knowledge about the Women's Health Initiative Randomized Controlled Trial 2002 (WHI-RCT) in Germany. DESIGN: During July 2003, telephone interviews were conducted with randomly selected women aged 45 to 60 years (N = 10,030; response 59.9%; completed interviews n = 6,007). They were asked about information sources regarding the WHI-RCT, and use of HT in conjunction with it. RESULTS: Most women stated that they knew about the WHI-RCT (88.6%), and those with high educational status appeared to have more information than those with middle or low educational status. Among informed women (uninformed excluded), 46.6% continued, 25.7% stopped, and 14.2% ceased use of HT before the WHI-RCT. The prevalence of lifetime use of HT was higher in West Germany (37.4%) than in East Germany (29.2%), the highest prevalence of use was in the group aged 55 to 59 years. The most common reason to continue HT was the benefit risk assessment by physicians (58%); the most common reason to stop HT were women's perceptions that the risks of HT exceeded the benefits (56%). If information was solely given by physicians, women were more likely to continue HT (60.4%), compared with mass media (46.1%), as a source of information. CONCLUSIONS: The survey demonstrates the impact of the WHI-RCT in Germany, and shows that both the media and advice given by physicians were important. Women who continued to use HT did so largely because of their physician's advice. Women who discontinued were mainly those who had a negative subjective perception about risk of HT. PMID- 16037756 TI - Effect of a combination of isoflavones and Actaea racemosa Linnaeus on climacteric symptoms in healthy symptomatic perimenopausal women: a 12-week randomized, placebo-controlled, double-blind study. AB - OBJECTIVE: To investigate the effects of a novel dietary supplement containing soy isoflavones and Actaea racemosa Linnaeus (formerly called Cimicifuga racemosa L.) on climacteric symptoms in healthy perimenopausal women. DESIGN: In a multicenter, randomized, placebo-controlled, double-blind study, 124 women experiencing at least five vasomotor symptoms every 24 hours were randomized to receive daily either a phytoestrogen-containing supplement (n = 60) or placebo (n = 64) for 12 weeks. The modified Kupperman Index and Greene Climacteric Scale, a visual analogue scale designed to measure quality of life and the daily number and severity of hot flushes, was used in the screening period and in weeks 6 and 12. Changes in these scores from baseline were calculated. RESULTS: At weeks 6 and 12, all scores in both groups had improved compared with baseline, though the overall difference in scores between the groups was not statistically significant. CONCLUSION: The supplement containing soy isoflavones and A racemosa L. had no statistically significant effect on climacteric symptoms in perimenopausal women experiencing at least five vasomotor symptoms per day. PMID- 16037757 TI - Vaginal estrogen therapy and overactive bladder symptoms in postmenopausal patients after a tension-free vaginal tape procedure: a randomized clinical trial. AB - OBJECTIVE: To evaluate whether the frequency of overactive bladder (OAB) symptoms increases in menopause patients after a tension-free vaginal tape (TVT) procedure, and to determine if topical estrogen therapy can help prevent these symptoms. DESIGN: After undergoing a preoperative assessment, enrolled patients were randomly allocated to receive TVT plus postoperative vaginal estrogen therapy (ET group) or TVT without adjunctive medical treatment (No ET group). The pre- and postoperative assessments included: acquisition of a urogynecologic history with standardized questions regarding urinary function (including a 10 grade visual analogue scale score), urogynecologic clinical examination, and urodynamic assessment. Follow-up assessments were performed at 1, 3, and 6 months after surgery. RESULTS: Fifty-six of 59 patients were evaluable; 28 received topic vaginal estrogen after surgery (ET group) and 28 did not receive adjunctive medical treatment (No ET group). The overall OAB syndrome rate in menopause patients (No ET group) was 7% (2 of 28 patients) at baseline and 32% (9 of 28 patients) 6 months after surgery (P = 0.04). At the 6-month follow-up assessment, the incidence of urinary urgency was 4% (1 of 28 patients) and 29% (8 of 28 patients) in the ET and No ET groups, respectively (P = 0.01). Differences in frequency and nocturia were not statistically significant. Analysis of the visual analogue scale scores revealed that at the 6-month follow-up assessment, urgency significantly improved in the ET group compared with the No ET group (0.23 +/- 1.0 vs 2.30 +/- 3.7, respectively; P = 0.02). CONCLUSIONS: The TVT procedure seems to increase the frequency of OAB syndrome in menopause patients. Vaginal estriol therapy significantly reduces symptoms of urinary urgency, has a high rate of patient satisfaction, and can be used to treat postmenopausal women for at least 6 months after a TVT procedure. PMID- 16037758 TI - Combination of soy protein and high dietary calcium on bone biomechanics and bone mineral density in ovariectomized rats. AB - OBJECTIVE: To determine if feeding soy in combination with a high-calcium diet would preserve bone mineral density and biomechanical bone strength to a greater extent than either soy or calcium alone. DESIGN: Rats were sham-operated (n = l0) and fed a control diet (AIN93G containing 0.2% calcium, 20% casein) or ovariectomized (n = 40) and randomized to one of the following diets (n = 10 per group): control, high calcium (2.5% calcium, 20% casein), soy (0.2% calcium, 20% soy protein), or soy plus high calcium (2.5% calcium, 20% soy protein) for 8 weeks. Bone mineral density of femurs and lumbar vertebrae 1 through 6 were measured by dual energy x-ray absorptiometry. Biomechanical strength properties of femurs and the fifth lumbar vertebrae were measured by three-point bending and compression, respectively. RESULTS: The dietary combination of soy and high calcium did not result in higher femur bone mineral density compared with other ovariectomized groups, and there were no differences in femur yield load or peak load among groups. In contrast, soy plus high calcium resulted in a higher (P < 0.05) vertebral bone mineral density compared with all other ovariectomized groups. Vertebral strength was preserved among rats fed either soy plus high calcium, soy, or high calcium whereas the ovariectomized group fed a control diet had lower (P < 0.05) vertebral strength than the sham-operated group. CONCLUSION: Bone mineral density of the lumbar spine was the only bone outcome that significantly benefited from the combination of soy and high calcium compared with soy or high calcium alone. PMID- 16037759 TI - Menopause symptoms and quality of life in women aged 45 to 65 years with and without breast cancer. AB - OBJECTIVE: To compare the prevalence of menopause symptoms, sexual activity, and quality of life in women with and without breast cancer. DESIGN: A cross sectional study using one group for comparison was conducted on women aged 45 to 65 years who had not received hormone therapy or tamoxifen during the last 6 months. Participants were recruited from the Menopause and Breast Cancer Outpatient Facilities. One hundred eighty-two women were included, 97 with breast cancer and 85 without breast cancer. Sociodemographic and clinical features and prevalence of menopause symptoms were assessed. The quality of life was assessed by the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) questionnaire. To compare sociodemographic and clinical features between groups, the Student's t test or Fisher exact test was used. Multiple logistic regression and linear regression models were used to control for potential confounding variables. RESULTS: The mean age of participants with breast cancer was 53.2 +/- 6.2 years, and the mean age of those without cancer was 57.8 +/- 4.9 years (P < 0.01). Age at menopause was 47.2 +/- 5.1 years and 47.4 +/- 4.9 years for women with and without breast cancer (P = 0.76), respectively. Approximately one-fourth of women with breast cancer and 4.7% of women without cancer were premenopausal (P < 0.01). The prevalence of menopause symptoms was similar between the groups. Women with breast cancer reported less sexual activity (51.5%) than women without cancer (62.4%) (P < 0.01). Quality of life scores were good in both groups. There was a significant difference regarding physical functioning, with a median score of 90 for the cancer group and 75 for the group without cancer (P < 0.01). CONCLUSION: The prevalence of menopause symptoms was similar in women with and without breast cancer. Sexual activity was less frequent in women with breast cancer. Quality of life was good in women from both groups, although women with breast cancer had the highest level of physical functioning. PMID- 16037760 TI - Vascular events in the Multiple Outcomes of Raloxifene Evaluation (MORE) trial: incidence, patient characteristics, and effect of raloxifene. AB - OBJECTIVE: To determine the incidence of arterial and venous thromboembolic (VTE) events, to determine the effect of raloxifene on the incidence of combined vascular (arterial and VTE) events, and to identify patient characteristics associated with these vascular events, in women participating in the MORE trial. DESIGN: In a post hoc analysis using MORE data, arterial, VTE, and combined vascular event rates were compared between participants receiving placebo (n = 2,576) and those receiving 60 mg/d of raloxifene (n = 2,557). Baseline characteristics were compared between those who did and did not experience an arterial event. The same analysis was performed for VTE events. RESULTS: Overall, during a mean follow-up time of 41 months, 178 women experienced an arterial event and 40 experienced a VTE event. In the placebo group, the incidence of arterial events exceeded VTE events by at least sevenfold. Raloxifene had no significant effect on the incidence of combined vascular events in the overall cohort (hazard ratio 0.95, 95% CI, 0.73-1.24). In a subset of women retrospectively determined to be at increased cardiovascular risk, raloxifene was associated with a lower incidence of combined vascular events (hazard ratio 0.63, 95% CI, 0.40-0.97). Baseline characteristics differed between those who did and those who did not experience an arterial event, but this was generally not the case for VTE events. CONCLUSIONS: Arterial events were more common than VTE events. The characteristics of women experiencing an arterial event differed from those experiencing a VTE event. Raloxifene had a neutral effect on the risk of combined vascular events in the overall population, and was associated with a reduced combined vascular event rate in women at increased cardiovascular risk. Additional studies are needed to confirm the effect of raloxifene on overall vascular outcomes. PMID- 16037761 TI - 17beta-estradiol exerts a beneficial effect on coronary vascular remodeling in the early stages of hypertension in spontaneously hypertensive rats. AB - OBJECTIVE: Estrogens may induce cardioprotective effects and prevent neointima formation in response to vascular injury in vivo. The present study evaluated the effect of 17beta-estradiol on myocardial arterial remodeling and on vascular mitogen-activated protein kinase expression in experimental hypertension. DESIGN: The experiments were performed in intact female spontaneously hypertensive rats (SHR), in SHR that were ovariectomized at 10 or 25 weeks of age, and in ovariectomized SHR that were supplemented with 17 beta-estradiol (OVX + E2, 1.5 mg every 8 weeks, subcutaneous pellets). RESULTS: At 18 weeks of age, in all myocardial arterioles and small arteries studied, we found significant increases in wall-to-lumen ratio in ovariectomized rats as compared with intact animals. 17beta-estradiol significantly reduced the wall-to-lumen ratio in OVX + E2 rats. Perivascular fibrosis of small coronary arteries was significantly increased by ovariectomy, and this effect was prevented by long-term treatment with 17beta estradiol. Phosphorylated extracellular signal-regulated kinase 1/2 significantly increased in mesenteric arteries from ovariectomized animals and this effect was prevented by 17beta-estradiol. Wall-to lumen ratio and perivascular fibrosis were significantly higher in older intact animals at 33 weeks of age. However, neither ovariectomy nor estradiol replacement had any effect on long-term hypertension induced vascular remodeling. CONCLUSIONS: These data suggest that estradiol may exert a beneficial effect by protecting the vasculature from hypertension-induced myocardial arterial remodeling in the early stages of hypertension, but not when chronic alterations are established after a long-term period of hypertension. PMID- 16037762 TI - Hot flushes during the menopause transition: a longitudinal study in Australian born women. AB - OBJECTIVE: To investigate factors associated with the presence, severity, and frequency of hot flushes. DESIGN: A 9-year prospective study of 438 Australian born women, aged 45 to 55 years and menstruating at baseline. Annual fasting blood collection, physical measurements, and interviews including questions about bothersome hot flushes in previous 2 weeks were performed. A "hot flush index" score was calculated from the product of the severity and frequency data. Data were analyzed using random-effects time-series regression models. RESULTS: A total of 381 women supplied complete data over the follow-up years. A total of 350 women experienced the menopause transition, of whom 60 (17%) never reported bothersome hot flushes. At baseline, women who reported hot flushes were significantly more likely to have higher negative moods, not be in full- or part time paid work, smoke, and not report exercising every day. Over the 9-year period of the study, variables significantly associated with reporting bothersome hot flushes were relatively young age (P < 0.001), low exercise levels (P < 0.05), low estradiol levels (P < 0.001), high follicle-stimulating hormone (FSH) levels (P < 0.001), smoking (P < 0.01), being in the late menopause transition (P < 0.001), or being postmenopausal (P < 0.001). In women reporting hot flushes, the hot flush index score increased as their FSH levels increased (P < 0.01), as they entered the late stage of the menopause transition (P < 0.001), and as they became postmenopausal (P < 0.05), and decreased with as their age (P < 0.001) and exercise level (P < 0.05) increased. Between-women analyses found that the hot flush index score was greater in women with higher average FSH levels over time (P < 0.05). CONCLUSION: Menopause status, FSH and estradiol levels, age, exercise level, and smoking status all contributed to the experience of bothersome hot flushes. PMID- 16037763 TI - The effects of progesterone, medroxyprogesterone acetate, and norethisterone on growth factor- and estradiol-treated human cancerous and noncancerous breast cells. AB - OBJECTIVE: Mitogenic growth factors from stromal breast tissue and estrogens are important in growth regulation of breast cells and may modify different responses dependent on the choice of progestogens. The effects of the acknowledged pharmacologically significantly different progestogens, progesterone, medroxyprogesterone acetate (MPA), and norethisterone, were investigated in breast cells treated with growth factor and/or estradiol, comparing normal and primary cancer cells for the first time. DESIGN: MCF10A (human epithelial, estrogen-receptor-negative and progesterone-receptor-negative normal breast cells) were incubated with progestogens at 1 microM and 100 nM for 7 days and epidermal growth factor, fibroblast growth factor, and insulin-like growth factor at 1 pM. HCC1500 (human estrogen-receptor-positive and progesterone-receptor positive primary breast cancer cells) were incubated in the same way with growth factors and/or estradiol at 100 pM. Cell proliferation rate was measured by the ATP assay, whereas cell death was measured by photometric enzyme immunoassay. Ratios of cell death to proliferation were calculated from these results. RESULTS: In combination with growth factors, MPA reduced the ratio in favor of a proliferative effect in MCF10A cells, followed by norethisterone. Progesterone had no significant effect on the growth factor response. In HCC1500 cells, MPA with growth factors increased the ratio favoring inhibition, norethisterone had a proliferative effect, and progesterone had no significant effect. In combination with estradiol, MPA increased the ratio to inhibition to the greatest extent, followed by norethisterone and progesterone at 1 muM. In combination with growth factors and estradiol, MPA and norethisterone both showed an inhibitory effect, whereas progesterone had no significant effect. CONCLUSION: Certain progestogens are able to induce the proliferation of benign or malignant human breast cells independently of the effects of growth factors and estradiol. Therefore, the choice of progestogen may be important in terms of influencing a possible breast cancer risk. PMID- 16037764 TI - Childhood cognitive ability and age at menopause: evidence from two cohort studies. AB - OBJECTIVE: To investigate whether poorer cognitive ability in childhood is associated with an earlier menopause. DESIGN: Two cohorts were included: a nationally representative British birth cohort study of 1,350 women born in March 1946 and followed up to age 54 years, and an Aberdeen cohort study of 3,465 women born in Aberdeen from 1950 to 1956 and followed up to age 44 to 50 years. Both cohorts had prospective information on childhood cognitive ability at age 7 or 8 years. RESULTS: In both cohorts, women with lower cognitive scores in childhood reached menopause earlier than women with higher scores. With follow-up of menopause to 49 years, the hazard ratio (HR) for one standard deviation of the cognitive score was 0.80 (95% CI, 0.72-0.90) in the Aberdeen cohort and 0.84 (95% CI, 0.73-0.97) in the older 1946 birth cohort. The effect was still evident in the 1946 birth cohort with follow-up of menopause to 53 years (HR = 0.87; 95% CI, 0.79-0.95). These ratios were weakly attenuated by adjustment for potential confounding effects of lifetime socioeconomic circumstances, parity, and smoking. CONCLUSIONS: The association between early cognitive ability and timing of menopause only partially reflects common risk factors, although residual confounding remains a possibility. Alternatively, early environmental or genetic programming may explain this association, perhaps through setting lifelong patterns of hormone release or causing transient hormonal changes at sensitive periods of development. These findings have implications for the interpretation of studies investigating an association between age at menopause and adult cognitive function. PMID- 16037765 TI - Effects of lidocaine with and without epinephrine on plasma epinephrine and lidocaine concentrations and hemodynamic values during third molar surgery. AB - Lidocaine with epinephrine is currently the most common local anesthetic agent used for impacted third molar surgery. The purpose of the present study was to define the adverse hemodynamic effects and plasma concentrations of lidocaine and epinephrine on 17 healthy patients during the impacted teeth operations. Arterial blood pressure (systolic blood pressure, diastolic blood pressure), heart rate, peripheral oxygen saturation range, and electrocardiography were measured by an automatic noninvasive pressure device and monitor. High-performance liquid chromatography was used to measure the changes of plasma concentrations of epinephrine and lidocaine from blood samples taken 5 different times during the operation. We concluded that lidocaine-epinephrine is effective local anesthetic and had no important adverse events in healthy patients during the third molar surgery. PMID- 16037766 TI - Fibromatosis of the tongue: a case report. PMID- 16037767 TI - Improving patient care through virtual oral medicine. PMID- 16037769 TI - The influence of the sagittal split osteotomy on the condylar cartilage structure and the subchondral vascularization of the temporomandibular joint: a preliminary study in goats. AB - OBJECTIVE: The concern that a sagittal split osteotomy of the mandibular ramus could compromise the subchondral vascularization and especially the venous outflow in the condylar area, which in turn could influence the structure of the condylar cartilage, prompted this preliminary study on goats. STUDY DESIGN: A sagittal split osteotomy was performed in the mandibular ramus at 1 side in each of 6 young adult goats. The contralateral side served as control. The animals were killed after different postoperative periods, ranging from 1 to 15 days. Histomorphometric analyses were performed after perfusion of the vascular system with India ink. CONCLUSION: Based on the observed cartilage thickening, the sagittal split osteotomy may influence the condylar cartilage in the first days postoperatively through a disturbance of the vascular supply. PMID- 16037768 TI - Risk of periodontal defects after third molar surgery: An exercise in evidence based clinical decision-making. AB - OBJECTIVE: The object of this study was to apply evidence-based principles to answer the question, What is the risk of having periodontal defects on the distal aspect of the mandibular second molar (M2) following third molar (M3) removal? STUDY DESIGN: To identify relevant articles for review, we completed a computerized literature search of Medline. The inclusion criteria for articles included prospective cohort studies or randomized clinical trials with follow-up periods of 6 months or more, and preoperative and postoperative measurements of periodontal probing depths (PDs) or attachment levels (ALs). RESULTS: Eight articles met the inclusion criteria. Overall, the reported mean changes in ALs or PDs on the distal of M2 6 months after M3 removal were clinically insignificant, ie, less than 2 mm. Six months after M3 removal, 52% to 100% of subjects had no change or improvement in ALs or PDs. Given periodontal disease present preoperatively, the number needed to treat (NNT) ranged from 3 to 10. Given healthy periodontal status preoperatively, 48% had worsening of their periodontal measures after M3 removal and the number needed to harm (NNH) was 2. CONCLUSION: Commonly, the second molar periodontal probing depth or attachment levels either remain unchanged or improve after third molar removal. For subjects with healthy second molar periodontium preoperatively, the indication for third molar removal needs to be evaluated carefully as these subjects have an increased risk for worsening of probing depths or attachment levels after third molar removal. PMID- 16037771 TI - Quality of dying in head and neck cancer patients: a retrospective analysis of potential indicators of care. AB - OBJECTIVE: The objective of this study was to evaluate the quality of the dying experience in a cohort of head and neck cancer patients by a retrospective analysis of potential indicators of care. STUDY DESIGN: The study design included retrospective case note analysis of previously evaluated factors considered important in the care of terminally ill patients and validated indicators of care obtainable from administrative data. The documented factors were analyzed to indicate specific elements of the quality of death and dying experience. RESULTS: Pain was a common symptom (84%) and was managed successfully in all patients, with 93% receiving opioids. Management of other symptoms, except neuropsychological problems, were satisfactory. Sixty-three percent of patients died in hospital and only 22% had a relative present at the time of death. Resuscitation status was documented in only 65% of the notes, though none were admitted to the intensive care unit or underwent resuscitation. Fifty-three percent of patients were admitted as an emergency in the last month of life and bleeding was the most common cause of admission. CONCLUSION: Valuable information can be obtained from a focused retrospective analysis. The indicators evaluated suggest that biomedical aspects of care are being recognized and addressed, although there is room for improvement. Psychosocial and spiritual aspects of care were not documented and must be taken into account to assess the "complete quality of dying" experience. PMID- 16037772 TI - Facial distortion secondary to idiopathic gingival hyperplasia: surgical management and oral reconstruction with endosseous implants. AB - Little information exists regarding the surgical and oral reconstructive options for the patient with idiopathic gingival hyperplasia, especially with regard to endosseous implant placement and recurrence of the disease. The authors report a case of severe gingival enlargement and bone resorption involving the maxillary and mandibular arches in a 21-year-old female causing distortion of facial features. The etiology could not be established but the condition was exacerbated during pregnancy, causing chewing, speaking, and esthetic problems. The hyperplastic tissue and all the teeth were removed surgically, followed by immediate placement of 2 endosseous implants. Restoration of esthetics and function with a maxillary denture and mandibular overdenture were completed 3 months later. No disease recurrence was noted at the 6-month postoperative interval. PMID- 16037773 TI - Dental management of patients with myasthenia gravis: a literature review. AB - Myasthenia gravis is a chronic neuromuscular disease characterized by muscular weakness and fatigability. Dental management of patients diagnosed with myasthenia gravis presents a challenge to the dental profession. A MEDLINE search of the English medical (limited to provision in dental care) and dental literature on myasthenia gravis and dental management published between 1975 and 2004 was conducted. In the dental literature, 12 articles were found, and only a few focused on myasthenia gravis and dental care. The purpose of this article was to review and summarize the clinical signs and symptoms associated with myasthenia gravis, highlighting the role of the dental profession in the process of the diagnosis and management of the oral and dental complications that might be associated with the disease, while avoiding myasthenic crisis. PMID- 16037774 TI - Current controversies in oral lichen planus: report of an international consensus meeting. Part 2. Clinical management and malignant transformation. AB - Despite recent advances in understanding the immunopathogenesis of oral lichen planus (LP), the initial triggers of lesion formation and the essential pathogenic pathways are unknown. It is therefore not surprising that the clinical management of oral LP poses considerable difficulties to the dermatologist and the oral physician. A consensus meeting was held in France in March 2003 to discuss the most controversial aspects of oral LP. Part 1 of the meeting report focused on (1) the relationship between oral LP and viral infection, with special emphasis on hepatitis C virus (HCV), and (2) oral LP pathogenesis, in particular the immune mechanisms resulting in lymphocyte infiltration and keratinocyte apoptosis. Part 2 focuses on patient management and therapeutic approaches and includes discussion on malignant transformation of oral LP. PMID- 16037775 TI - Tuberculosis affecting the oral cavity in Brazilian HIV-infected patients. AB - OBJECTIVE: Tuberculosis (TB) affecting the oral cavity is a very rare disease. We reviewed the medical data of 18 HIV(1)-infected patients referred to the Stomatology Group, Ear, Nose, and Throat Department, Sao Paulo University Medical School, who had a diagnosis of oral tuberculosis, between 1991 and 2002. This retrospective study aimed to analyze the relationship between oral tuberculosis and HIV infection. STUDY DESIGN: The clinical records of the patients were reviewed to note age and gender, symptoms, and site and type of the lesions in the oral mucosa. Results of CD4+ T-lymphocyte count, sputum analysis, tuberculin skin test, histopathologic characteristics, cultures of the specimens (that were taken during the biopsy), chest x-ray findings, and the type of treatment they received were also reviewed. RESULTS: The most affected sites were palate and tongue. All patients had culture confirming tuberculosis. CONCLUSION: Patients with HIV and nonhealing oral lesions or papillomatous masses in the oral mucosa should be screened for the presence of pulmonary tuberculosis. PMID- 16037776 TI - Primary oral sarcoidosis preceding Lofgren's syndrome. PMID- 16037778 TI - Oral plasmablastic lymphoma in an HIV-negative patient: a case report and review of the literature. AB - Plasmablastic lymphoma is an HIV-associated non-Hodgkin's lymphoma that primarily affects the oral cavity and jaws. The purpose of this report is to describe the first case of plasmablastic lymphoma occurring in an HIV-negative, nonimmunocompromised individual, and to review the histopathologic and immunohistochemical phenotype of this lymphoma. Histopathologically, our case exhibited a dense, diffuse lymphocytic infiltrate of noncohesive large lymphocytes with plasmacytoid features. Immunohistochemical analysis revealed positivity for the B-cell marker CD79a, VS38c, Epstein-Barr virus latent membrane protein (LMP), immunoglobulin G (IgG), and lambda light chain restriction. Neoplastic cells were negative for leukocyte common antigen, CD20, CD3, CD10, CD138, Bcl-2, Bcl-6, desmin, actin, EMA, S-100, HMB45, Alk-1, HHV8, IgA, IgM, and cytokeratin. The features of this rare disease are summarized based on a comprehensive review of the epidemiologic, clinical and immunohistochemical findings of previously reported cases. PMID- 16037779 TI - Diagnosis and classification of mandibular osteomyelitis. AB - To establish a unified classification system for mandibular osteomyelitis, various diagnostic terms were critically assessed and clinicopathologic findings of the lesions were carefully reviewed. We recommend classifying mandibular osteomyelitis into bacterial osteomyelitis and osteomyelitis associated with the synovitis, acne, pustulosis, hyperostosis, and osteitis (SAPHO) syndrome. Other diagnostic terms were excluded because they were not appropriate for classification. Diagnostic criteria for bacterial osteomyelitis are suppuration and osteolytic change. The lesions are easily cured by antibiotic treatments. Mandibular osteomyelitis in SAPHO syndrome is characterized by nonsuppuration and a mixed pattern on radiography, with solid type periosteal reaction, external bone resorption, and bone enlargement. The presence of osteomyelitis in other bones, arthritis, or skin diseases (palmoplantar pustulosis, pustular psoriasis, and acne) strongly suggests this syndrome. Antibiotic therapy is usually ineffective and the symptoms of SAPHO syndrome are often persistent. PMID- 16037780 TI - Utility of magnetic resonance cisternography using three-dimensional fast asymmetric spin-echo sequences with multiplanar reconstruction: the evaluation of sites of neurovascular compression of the trigeminal nerve. AB - OBJECTIVE: To evaluate the utility of magnetic resonance (MR) cisternography using 3-dimensional (3D) fast asymmetric spin-echo (FASE) sequences with multiplanar reconstruction (MPR) for detection of the sites of neurovascular compression (NVC) in patients with trigeminal neuralgia. STUDY DESIGN: Both MR cisternography with 3D-FASE sequences and MR angiography (MRA) were performed on 150 patients with clinical signs and symptoms that suggested trigeminal neuralgia. Results from the original MR cisternography with 3D-FASE sequences, the original MRA, and 4 reformatted images were used for interpretation. Images with inversion between black and white were used from the MR cisternography with 3D-FASE sequences to evaluate NVC. A diagnosis of NVC was made from the presence of vascular contact with the trigeminal nerve at the root entry zone (REZ) and the nature of the involved vessels. For the patients with NVC detectable on 3D FASE or MRA images, the relationship between the clinically manifested regions and the NVC sites was also evaluated. RESULTS: Of the 150 patients evaluated, 89 were deemed to have NVC. Of these 89 patients, 3 underwent surgery that identified the artery that was involved with the trigeminal nerve. The correlation between the clinically manifested regions and the NVC sites was significantly detectable using both 3D-FASE images and MRA in 89 patients with detectable NVC. The correlation coefficient using 3D-FASE imaging was a little higher than that using MRA. Of the 61 patients deemed not to have NVC, 6 were found to have brain tumors that invaded the root entry zone of the trigeminal nerve. However, the remaining 55 patients had no identifiable cause for trigeminal neuralgia. NVC was found in the asymptomatic side in 27 (18%) of the 150 patients with trigeminal neuralgia using MR cisternography with 3D-FASE sequences. CONCLUSIONS: The technique of MR cisternography using 3D-FASE sequences with MPR is more accurate and useful than MRA for detection of the site of NVC in patients with trigeminal neuralgia. Patients with trigeminal neuralgia should be further examined using MR imaging to rule out a brain tumor. Radiologists should understand that a few subjects with no symptoms could show NVC with MR cisternography. PMID- 16037781 TI - Visual assessment of the mandibular cortex on panoramic radiographs to identify postmenopausal women with low bone mineral densities. AB - OBJECTIVE: The purpose of this study was to determine whether simple visual estimation of the mandibular inferior cortex width on panoramic radiographs is useful for identifying postmenopausal women with low skeletal bone mineral density (BMD). STUDY DESIGN: Panoramic radiographs were made on 100 women aged 50 84 years who also had BMD assessment of the lumbar spine and femoral neck by dual energy x-ray absorptiometry. The panoramic images were examined twice by 4 observers to evaluate the thickness of the mandibular inferior cortex. RESULTS: Intraobserver agreement varied from moderate to almost perfect. Interobserver agreements varied from fair to almost perfect. Overall mean sensitivity, specificity and positive predictive value in identifying women with low BMD were 54.7%, 82.0%, and 72.5% in the lumbar spine and 51.8%, 82.2%, and 75.7% in the femoral neck, respectively. CONCLUSIONS: Our results suggest that simple visual estimation of the mandibular inferior cortex on panoramic radiographs may be useful for identifying postmenopausal women with low BMD. PMID- 16037782 TI - Streptococci from root canals in teeth with apical periodontitis receiving endodontic treatment. AB - OBJECTIVES: The object of this study was to investigate the diversity among streptococcal species isolated from root canals in conjunction with endodontic therapy and to characterize their production of extracellular proteins. STUDY DESIGN: Consecutive root canal samples (RCS) taken as bacteriological controls during root canal treatment of teeth with apical periodontitis were analyzed in a total of 100 clinical cases. Bacteria were isolated and classified by selective media and gas liquid chromatography. Streptococcal strains were identified by carbohydrate fermentation, hydrolysis of aesculin/arginine, and production of enzymes. Releases of extracellular proteins by streptococci and Enterococcus spp in fluid culture media were examined with SDS-PAGE and 2-dimension gel electrophoresis (2 DE). Extracellular proteins produced were quantified and qualitatively analyzed. Specific proteins were targeted with Western immunoblot assays. Comparisons were made with type strains. RESULTS: Of a total of 241 bacterial strains recovered in the first samples submitted, Streptococcus gordonii, S anginosus, and S oralis were the most frequently isolated streptococci. In 49 of 89 resubmitted samples showing bacterial growth, S gordonii and S oralis still predominated among streptococci. Other common bacterial isolates were Enterococcus spp, Lactobacillus paracasei, and Olsenella uli. Quantitative and qualitative differences in extracellular protein production were observed among clinical isolates and laboratory streptococcal strains. In similar conditions for growth, S intermedius, S anginosus, S oralis, and S gordonii were strong producers of extracellular proteins (>3.0 microg/mL), while Enterococcus spp and S mutans were weak. Whole cell protein extracts showed a different profile from that of extracellular proteins. The chaperone protein DnaK was recognized to be produced extracellularly by S gordonii, S oralis, S anginosus, and S parasanguis. CONCLUSIONS: Being strong producers of extracellular proteins and by virtue of common presence in teeth undergoing endodontic therapy, S gordonii, S anginosus, and S oralis may be of pathogenic significance in posttreatment apical periodontitis. PMID- 16037783 TI - Nonsurgical retreatment of teeth with periapical lesions previously managed by either endodontic or surgical intervention. AB - OBJECTIVE: The purpose of this study was to evaluate the outcome of nonsurgical retreatment of teeth with periapical lesions, which had been previously managed by either endodontic or surgical intervention, and to identify factors that might influence the prognosis. STUDY DESIGN: Seventy-five endodontically and 11 surgically treated teeth with persisting periapical lesions ranging in size from 2 to 11 mm in diameter were included in this study. The teeth were nonsurgically retreated, using calcium hydroxide as intracanal medicament, and were followed for a period of 2 to 8 years. RESULTS: Clinical and radiographic assessment of retreatment showed that complete healing for all cases was 61.6% with an additional category of incomplete healing of 14%; 24.4% failed. The size of the periapical lesions and previous surgical treatment had marginally negative influences on the prognosis, but the differences were not statistically significant. CONCLUSION: The favourable results of this study demonstrated that nonsurgical root canal retreatment is an alternative to surgical intervention of postendodontic or postsurgical failure. PMID- 16037784 TI - Decrease in the fatigue resistance of nickel-titanium rotary instruments after clinical use in curved root canals. AB - OBJECTIVE: The changes in fatigue resistance of nickel-titanium rotary ProFile instruments after clinical use for shaping 10 curved molar root canals were evaluated in this study. STUDY DESIGN: Twenty-five sets of files #20, #25, and #30 and tapers .04 and .06 were divided into 2 groups, one with 10 sets of new files that were tested in a fatigue test bench device as a control. The other, experimental group, with 15 sets of clinically used files, was tested in the same device. The Student t test was employed to compare mean values of the measured parameters. RESULTS: A statistically significant decrease in the number of cycles to failure was determined for the clinically used files, as compared with the new ones. The fracture point was the same for all files tested. CONCLUSIONS: The clinical use of ProFile instruments for shaping curved canals reduces their fatigue resistance. PMID- 16037785 TI - Our journal reaches an important milestone. PMID- 16037786 TI - Advances since the paper by Zander and Glass (1949) on the pursuit of healing methods for pulpal exposures: historical perspectives. AB - Clinical observations and experimental studies in humans and laboratory animals have demonstrated that healing and repair of pulpal exposures by caries, trauma, or iatrogenic causes are possible with a variety of wound treatment methods. Yet clinical trials have shown that predictable long-term pulp tissue preservation may be an elusive goal and has led to doubts about pulp capping and pulpotomy as valid clinical procedures. Nevertheless substantial knowledge has accumulated over the years on the mechanisms and the treatment factors that are important to promote/support continued vital pulp functions. This article highlights some key contributions to our current knowledge base, which have come to light during the more than 50 years since a pioneering experimental study by Zander and Glass was published in the Triple O journal. PMID- 16037787 TI - A review and update of new methods for immobilization of the mandible. AB - In 1948, Dr Kurt H Thoma, a leading authority of the day, published a paper on new methods for immobilization of the mandible in the first issue of this Journal. He reviewed the state of the art for management of patients with fractures of the mandible. By reviewing the paper now we can see how difficult to treat some of the injuries he saw were. This paper assesses his patient management in light of today's knowledge and experience. It discusses changes in therapy that have occurred during this over fifty-year time span. Although many things have changed since then, what has not changed is our desire to provide the best care for our patients using currently available scientific knowledge and clinical evidence. PMID- 16037788 TI - Viruses as agents of disease. 1948. PMID- 16037789 TI - The prevention of virus disease. 1948. PMID- 16037790 TI - Pleiotropic mechanisms of virus survival and persistence. AB - Viruses are enormously efficient infectious agents that have been implicated in causing human disease for centuries. Transmission of these pathogens continues to be from one life form to another in the form of isolated cases, epidemics, and pandemics. Each infection requires entry into a susceptible host, replication, and evasion of the immune system. Viruses are successful pathogens because they target specific cells for their attack, exploit the cellular machinery, and are efficient in circumventing and/or inhibiting key cellular events required of survival. This article reviews some of the advances that have taken place in human virology in the past 50 years, emphasizing mechanisms that contribute to, and are involved with, virus survival and persistence. PMID- 16037791 TI - Eosinophilic granuloma of bone: case report of eosinophilic granuloma of mouth (jaws, gums, and palate) with simultaneous fistula in ano. 1948. PMID- 16037792 TI - Langerhans cell histiocytosis: current insights in a molecular age with emphasis on clinical oral and maxillofacial pathology practice. AB - Langerhans cell histiocytosis (LCH) commonly involves the oral and maxillofacial region, and comes to the attention of dental practitioners when a patient presents with orofacial pain and a bony or soft tissue lesion. This is a relatively rare entity, which has made it difficult to investigate the clinical, biologic, and molecular aspects of the disease. Treatment protocols are not well defined, particularly in adults. During the past decade, the Histiocyte Society has formulated various LCH categories, based on risk stratification, and treatment protocols for the pediatric population. Adult trials are currently available through the Histiocyte Society. Although there has been considerable controversy, the neoplastic nature of LCH has been established by demonstrating clonality. LCH symptoms and the development and persistence of LCH lesions have been ascribed to a "chemokine/cytokine storm" due to autocrine and paracrine mechanisms. Discovery of biologic, cytogenetic, and molecular abnormalities in LCH have already affected treatment by providing novel therapeutic targets. PMID- 16037793 TI - New methods for immobilization of the mandible. 1948. PMID- 16037794 TI - X-ray evidence of mechanical trauma. 1959. PMID- 16037795 TI - Imaging of maxillofacial trauma: evolutions and emerging revolutions. AB - The aim of diagnostic imaging for maxillofacial trauma is to provide additional information that can positively influence medical or surgical patient management. Current advances in diagnostic imaging have come from the confluence of 3 driving forces: (1) the demand from clinicians to enhance and expand their diagnostic abilities; (2) the development of new theoretical concepts by basic scientists; and (3) the application of concepts by engineers and manufacturers to provide increasingly sophisticated imaging capabilities. The role of imaging within the health care environment is, however, also buffeted by the complex, sometimes competing, interactions of external social, political, economic, and technological pressures at the national, regional, and local levels. The purposes of this review are to provide a perspective on current imaging modalities used for maxillofacial trauma and to provide an insight into the influences, both technologic and external, on future developments and applications. PMID- 16037796 TI - The healing of phenolized pulp exposures. 1949. PMID- 16037797 TI - [The role of the immune system in the pathogenesis of atherosclerosis]. AB - This paper is concerned with intracellular infections such as herpes virus, cytomegalovirus and Chlamydia pneumonia which may be implicated in the pathogenesis of atherosclerosis. These viral and bacterial pathogens are regarded as potential causative factors in the pathogenesis of atherosclerosis. The authors support the hypothesis that these infections may induce endothelial injury. The paper reviews both direct and indirect immune system-mediated effect of the viruses indicated on the disease process and the potential role of killer cells in endothelial injury. PMID- 16037798 TI - [Predictive significance of insulin resistance and metabolic syndrome for assessment of the degree of endothelium disfunction and early signs of atherosclerotic lesion]. AB - The contribution of the clinical or laboratory risk factors associated with the metabolic syndrome to the changes in peripheral vascular reactions and to the natural history of atherosclerosis has not been established until now. The aim of the work was to study interrelations between the risk factors entering the symptom complex of the metabolic syndrome, to assess their impact on endothelium dependent vasodilatation and constrictor component of vascular reactivity as well as on the change in the thickness of the intima-media complex (TIMC) of the common carotid artery (CCA). The study accrued 122 practically normal men aged 35 to 50 years. Stepwise multiple regression analysis has established interrelations between the disorder of vasoregulating endothelium function and the intensity of atherosclerotic lesion according to the maximal value of the TIMC of the CCA, glucose concentration 120 min after glucose load, IMT, the signs of arterial hypertension. The data have been obtained that confirmed the hypothesis of the role played by endothelium dysfunction manifesting by the disturbance of vasoregulating function, as an early marker of atherosclerosis. Carbohydrate metabolism, tissue sensitivity to insulin and the presence of arterial hypertension were most significant predictive factors of the TIMC of the CCA and disorder of arterial wall function. PMID- 16037799 TI - [The loss of work fitness and the course of invalidism in patients with limb vessel lesions]. AB - The growth of the sick rate of limb peripheral vessels associated with a severe outcome (trophic ulcers, amputation) exerts an appreciable effect on the lowering of quality of life in patients. This manifests by the prolonged loss of work fitness, change of the habitual occupation and disability establishment. Objective analytical information on this problem will be of help in the delineation of the tendencies in this direction and potential approaches to the prevention of social losses. The present work is based on an analysis of 2115 statements of medicosocial expert evaluation (MSEE) of invalids suffering from diseases of limb vessels, performed over recent 8 years in the Altai region. The decisions made by the MSEE were based on the results of the clinical examination of patients using the current diagnostic modalities (ultrasonography, duplex scanning, angiography, etc). It has been established that among persons who had undergone MSEE, over the half (64.1%) were under 60 years, i.e. in the age of work fitness. It is noteworthy that the overwhelming number of invalids were men (83%) and workers (84.2%). As for special vascular pathologies, the majority of patients presented with obliterative arterial diseases (OAD) of the lower limbs, accounting for 76.3% whereas patients with venous pathology ranked second in number (15.9%). The highest severity of invalidism (groups I and II) was also recorded in OAD (77.5%), especially in atherosclerosis obliterans (AO) which accounted for 84%. Of note, these diseases were marked by no tendency toward reduction of their incidence. The time of temporary disability (from 3 to 9 months) was also most frequently recorded in OAD of the limbs. In OAD, the temporary or persistent loss of work fitness were caused by critical ischemia and amputations whereas in venous pathology, namely in varicosity and post thrombophlebotic syndrome, the cause was progressing CVI complicated by trophic ulcers. On the whole, the lack of changes in the lowering of the number of invalids due to the given pathology evidences the unsatisfactory results of these patients' rehabilitation and the high socioeconomic tension determined by considerable treatment expenses and the high number of the disabled. Approaches to the escape from such a situation should be looked for in the early mass screening diagnosis of vascular lesions, early drug and surgical treatment and in the refinement of the system of rehabilitation prophylactic medical examination. PMID- 16037800 TI - [Combined bolus magnetic resonance angiography and two-dimensional time-of-flight magnetic resonance angiography in patients with occlusive diseases of lower limb arteries]. AB - AIM: The purpose of the work was to assess the diagnostic significance of combined bolus magnetic resonance angiography (MRA) and two-dimensional time-of flight MRA of lower limb arteries as compared to radioangiography in patients with occlusive diseases of lower limb arteries. MATERIAL AND METHODS: Twenty-six patients with the symptoms of lower limb ischemia of varying severity were provided bolus MRA with prolonged manual injection of contrast substance and transposition of the MR-tomograph table. Additionally, all the patients underwent two-dimensional time-of-flight (2D TOF) MRA of the leg arteries. RESULTS: The statistic processing of bolus MSA for visualization of the pelvic segment has demonstrated its high sensitivity and specificity (97.74% and 92.31% respectively). On visualization of the femoral artery segment the sensitivity of this modality accounted for 97.57%, the specificity for 94.57%. In patients with severe ischemia, visualization of the leg arteries was difficult. Consequently, the specificity of the modality accounted for 44.58%. However, when this technique was combined with two-dimensional time-of-flight MRA of the leg arteries, the sensitivity and specificity of the modality reached 96.38% and 91.92% respectively. CONCLUSION: Bolus MRA of lower limb arteries is marked by the high sensitivity and specificity for visualization of the arteries of the iliac and femoral segments. To visualize the leg arteries in patients with severe forms of lower limb ischemia, this modality should be supplemented by two dimensional time-of-flight MRA. PMID- 16037801 TI - [Cerebral venous hemodynamics in chronic disorders of cerebral circulation]. AB - The aim of the work was to assess extra- and intracranial venous hemodynamics in patients with circulatory disorder-induced encephalopathy (DE). Altogether 114 DE patients were examined. There were 46 women and 68 men aged 43 to 78 years (mean age 59.6+/-12.5 years). As dependent on the clinical manifestations the patients were distributed into groups: stage 1 DE was present in 36 patients, stage 2 DE in 47>> and stage 3 DE was identified in 31 patients. 82.78% of the examined had arterial hypertension (AH), the mean standing of which accounted for 9.7+/-7.2 years. The control group accrued 36 practically normal persons aged 36 to 62 years (mean age 47.6+/-11.3 years). All the patients were provided standard neurologic examination, magnetic resonance tomography (MRT) of the brain with venography of the brachiocephalic veins and venous sinuses of the brain, Color Doppler Imaging of the extracranial vessels, and transcranial Doppler. The patients complained primarily of headache, dizziness, instability and staggering on walking, memory and work fitness decrease, and irritability. Atherosclerotic plaques which were primarily homogeneous (types IV and V according to the classification by Geroulakos et al., 1993) were identified in the carotid arteries in 76 (62%) patients. In 48 (42%) patients, stenoses were bilateral. Hemodynamically significant (>50%) stenoses were present in 42 (34%) persons. In most cases, the patients showed dilatation of the jugular veins and a mean reduction of the flow intensity to 14+/-8.1 cm/s as compared to the control group (20.6+/-11.3 cm/s). The tendency toward flow intensity lowering associated with its phasic nature disorder was particularly well-defined in patients with stage 3 DE and a long-term history of AH. On examination of the parameters of cerebral venous circulation the patients with stage 1 DE tended to the rise of the linear flow velocity (LFV) in the basal veins of Rozenthal and in the direct sinus. However, no significant changes in the PI parameters were recorded. In the patient group with stages 2 and 3 DE, there was an appreciable rise of the LFV in the deep veins in the presence of a remarkable PI lowering (the flow velocity in the vein of Rozenthal 21.8+/-7.2 cm/s in stage 2 DE, and 24.4+/-7.2 cm/s in stage 1 DE). In 87 (79%) cases, MRT revealed the signs of diffuse ischemic lesion of the brain. Fifty-five (48%) patients were diagnosed to have leukoarayos whereas in 48 (42%) cases, there were identified multiple lacunar infarctions, primarily of the deep cerebral segments. Ten (8%) patients demonstrated type 1 Arnold Chiari abnormalities -- hypoplasia of the large cerebral cistern and 4 patients had porto-cerebellar atrophy (megacysterna magna). Analysis of the MRV revealed, in the majority of cases (in 67 or 59%), developmental abnormality of the drainage system of the brain. Thus, 42 (37%) patients were diagnosed to have hypoplasia of one of the transverse sinuses (of the right one in 23 cases and of the left one in 19 cases); 17 (15%) persons had aplasia of the transverse sinus. Eight patients showed hypoplasia of the sigmoid sinuses (of the right one in 5 cases and of the left one in 3 cases). In all the cases of developmental abnormalities of the venous sinuses, there was a compensatory dilatation of the contralateral sinus and in some cases, there were visualized the upper and lower sinuses, the identification of which in health is difficult. So, atherosclerosis of AH-induced lesion of the brachiocephalic arteries interferes with the action of the physiological "arteriovenous pump" thereby provoking venous congestion. Progression of the process is associated with depletion of the compensatory adaptive potentialities of the collateral venous outflow which (especially in concomitant developmental abnormality in the region of the posterior cranial fossa and venous sinuses) favours aggravation of venous circulatory distress, the rise of the CSF pulse pressure and the emergence of benign intracranial hypertension and hydrocephalus followed by cerebral atrophy. PMID- 16037802 TI - Temporary placement of the cava filter to the suprarenal segment of the inferior vena cava. AB - This paper describes successful results of temporary placement of the Russian removable cava filter "Zontik" to the suprarenal segment of the inferior vena cava for the time of performing cesarean section and for the short-term postoperative period in a patient with late pregnancy, venous thrombosis of the lower limbs and pulmonary thromboembolism. PMID- 16037803 TI - [Roentgenendovascular stenting (stent-graft) of the external iliac artery for elimination of post-traumatic arteriovenous anastomosis]. AB - Presented herein is a case report concerning successful treatment of the post traumatic pathological anastomosis between the left external iliac artery and iliac vein using the stent-graft Jostent-graft. The anastomosis emerged after gunshot injury to the abdominal cavity. PMID- 16037804 TI - [Repeated surgical treatment of varicosity: the causes and specific features]. AB - The paper is concerned with repeated surgical treatment for varicosity. The studies were carried out of the efficacy of surgical treatment at the specialized angio-surgical and general surgery hospitals. It has been discovered that after treatment at the specialized hospitals the patients applied for repeated surgical assistance 6 times less frequently. It is shown that the incompetent perforating veins provoke the recurrence of the symptoms of varicosity. An original technique for their obliteration is offered using highly intensive laser radiation (HILR) under permanent ultrasound guidance. The optimal parameters of HILR for venous obliteration were worked out in animal experiments. The experimental data were confirmed by microscopy and electron scanning microscopy. The clinical observations have demonstrated the high efficacy and safety of the treatment method offered. PMID- 16037805 TI - [The modern concept of the tactics of conservative and combined treatment of limb lymphedema]. AB - The paper is concerned with the general problems of the tactics and stages of conservative and combined treatment of limb lymphedema using compression therapy techniques. The data presented herein cover both our own studies and the reported materials which are grouped so as to understand the basic principle of the concept of combined therapy for lymph edema (CTLE). The treatment presumes the use of two main variants in the form of CTLE as an independent method of conservative treatment of lymphedema and CTLE as a program coupled with surgical operations. The treatment policy for lymphedema involves two basic stages: hospital and prophylactic. The hospital program of the CTLE includes technologies acting on different pathogenetic components of lymphedema. In primary lymphedema, the hospital treatment allows to minimize lymphedema to 36% and in secondary to 4 4% of the initial parameters. During the prophylactic period, edema regression is appreciably delayed or does not take place at all. The next stage aimed at edema decrease is followed by a course of hospital CTLE which, as dependent on lymphedema severity, is carried out 1-4 times a year. The main principle of a current approach to the treatment of lymphedema is based on the concept of multimodality independent or combined treatment. The accuracy of this principle observance predetermines the efficacy of the results obtained. PMID- 16037806 TI - [Specialized outpatient care for patients with initial forms of lower limb chronic venous insufficiency]. AB - Authors present 7-year follow up of 567 patients with initial forms of chronic venous insufficiency (CVI) of lower extremities (442 patients with C-1 class and 125 patients with C-2 class). The analysis of individual clinical and ultrasonic data has shown wide variability and the lack of specificity in CVI symptomatology. Ultrasonic investigation revealed veno-venous reflux in 74.6% of patients with C-1 class. Complex outpatient care adhered the principles of minimal invasiveness and radicality, along with individual approach to surgical correction and drug therapy. Early (within 3 weeks) and late (3 and 12 months) outcomes were assessed, including trends in CVI subjective symptoms and objective evaluation of changes in superficial venous system. One year later full clinical and esthetic effect was achieved in 91.4% of C-1 class patients and in 75.8% of C 2 class patients. PMID- 16037807 TI - [Surgical method for correction of absolute valvular insufficiency in post thrombophlebotic disease of the lower limbs]. AB - A new technique has been worked out for intravasal autovenous valvuloplasty. It is based on the "ink-pot that does not spill" principle with creation of two cusps and is used in the treatment of patients with post-thrombophlebotic disease of the lower limbs induced by absolute valvular incompetence of the great veins. Criteria are offered for assessment of the efficacy of the technique for formation of venous valves. Altogether 54 operations were performed including those provided to 29 patients (the main group) operated on according to the technique proposed and those in 25 patients (control group) operated on the basis of the known technique reported in the literature. In 51 patients, these operations were accomplished in combination with different variants of procedures on the communicating and saphenous veins of the lower limbs. The time of some follow ups was as long as 7 years. The beneficial short-term results were obtained in 96.5% of cases whereas the long-term outcomes were positive in 92% of the basic group patients. PMID- 16037808 TI - Carotid thromboses in the early postoperative period after carotid endarterectomy. AB - The paper analyzes 10 cases of internal carotid artery (ICA) thrombosis, which occurred after 635 carotid endarterectomies (CEA), carried out from January 1997 to July 2004. CEA procedures included 346 (54.5%) open CEAs with PTFE patch angioplasty and 289 (45.5%) eversion CEAs. Patients with thromboses in the reconstructed area (n=10) had the profiles of comorbidities and cerebrovascular insufficiency grade similar to the total CEA group (p>0.1). Differences concerned the higher rate of atrial fibrillation cases (10%), diabetes mellitus (30%) and contralateral ICA occlusions (20%) in the group of thromboses (p>0.1). Among these 10 patients, 5 underwent eversion CEA (1.44% of the total eversion CEA group) and 5 - open CEA with patch angioplasty (1.73%). In 8 patients thromboses manifested as local neurological symptoms in the area supplied by the operated carotid artery. One patient demonstrated the progression of general cerebral symptomatology, while in the tenth patient thrombosis was accompanied with cerebral coma. Urgent ultrasonography in 7 patients failed to detect a blood flow in ICA, that proved the diagnosis of ICA thrombosis; in 3 patients ultrasonography showed a mural thrombosis. Nine patients underwent reoperation in emergency, while for the tenth patient an intervention was contraindicated due to the thrombosis of middle cerebral artery, unstable clinical state (cerebral coma) and progression of neurological symptomatology. PTFE arterial graft was implantedin 7 reoperated patients with ICA thrombosis, thrombectomy from ICA was carried out in 1 patient and in another one patient a thrombectomy from reconstructed segment with PTFE patch angioplasty of the arteriotomy defect was fulfilled. In 3 reoperated patients a complete resolution of neurological symptoms was evident 24 hours after intervention, in 2 patients neurological symptomatology regressed over 2-4 weeks. In another 3 reoperated patients local neurological symptoms persisted; 1 patient demonstrated postoperative progression of neurological symptomatology and coma with fatal outcome. The last patient, for whom a reoperation was contraindicated, died due to aggravation of cerebral coma. Dynamic follow-up in early postoperative period after CEA, especially in the first 6 hours, as well as an urgent duplex scanning of the reconstructed area in case of neurological symptoms development, provides timely diagnosis of postoperative thrombosis and the success of reoperations. Adjustments to patient's cardiac status and appropriate intraoperative heparinization can decrease the risk of this complication. An urgent reoperation aimed at the restoration of carotid blood flow, immediately after the diagnosis of thrombosis (provided the absence of deep coma), promotes a complete or partial regression of neurological symptoms and helps to prevent a progression of cerebrovascular disturbances. PMID- 16037809 TI - Risk factors and prevention of injuries to the cranial nerves in reconstructive surgery of the carotid arteries. AB - Reconstructive operations on aortic arch branches is the most effective approach to prevention of acute and chronic disorders of cerebral circulation. Iatrogenic injuries to the cranial nerves worsen the early end, particularly, the late postoperative period, decrease the quality of life and the social status of patients who had undergone carotid reconstructions. The aim of the study was to improve the short- and long-term results of reconstructive operations on the carotid arteries by means of minimizing the incidence and severity of iatrogenic injuries to the cranial nerves. The study accrued 149 patients undergoing operations on the carotid arteries for atherosclerosis or pathologic tortuosity. Of these 82 patients forming the control group were examined for the incidence and character of injuries to the cranial nerves. Neuropathy of the cranial nerves (CN) was identified in 16 (19.5%) patients (7 patients had injuries to the hypoglossal nerve, 3 to the facial nerve, 5 to the vagus; one patient presented with coexistent injury to the glossopharyngeal and pharyngeal branches of the vagus). The clinically and statistically significant risk factors of injuries were: minor surgical experience, the high loop of the internal carotid artery (ICA), lengthy atherosclerotic stenosis greater than 2 cm, diabetes mellitus, intraoperative trauma of the area of the cranial nerves, high mobilization of the ICA, the lack of visualization of pairs X and XII of the CN, intraoperative bleeding, intersection of the superior radix of the deep cervical loop, edema and hematoma of the neck in the postoperative period, and early unscheduled reoperations. One month later the cumulative stability of cranial dysfunction accounted for 62.5%, after 3 months it accounted for 43.8%, after 6 months for 31.2 , after 9 months for 18.8%, and after 12 months for 6,2%. In patients with injury to the CN, analysis of the quality of life made in the late postoperative period revealed its lowering with respect to all the constituents of mental and physical health. Multimodality prevention of CN injuries was carried out in 67 basic group patients and was aimed at exclusion or abatement of the intensity of the impact of the removable risk factors. The proposed measures made it possible to minimize the incidence of CN injuries from 19.5 to 4.5% and to appreciably improve the quality of life of operated patients. PMID- 16037810 TI - The long-term results of surgical treatment of ascending aortic aneurysms. AB - The paper summarizes an experience with surgical treatment of ascending aortic aneurysms, gained at the Russian Scientific Center of Surgery, RAMS. Ninety-two patients operated on for the complicated and uncomplicated forms of ascending aortic aneurysms were entered into the study. A comparative analysis was made of the long-term results of different types of surgical interventions. The authors demonstrate the advantages of operations using valve-containing conduits and of operations which consisted in graded intima-sparing resection of aortic aneurysm and eco-prosthetics. The survival of patients over the period as long as 12 years after operation accounted for 86%. PMID- 16037811 TI - Combined operations for multilevel lesions of the aortoiliac and femoropopliteal segments. AB - This paper describes the results of the treatment of 53 patients with combined arterial lesions of the iliofemoral and femoro-popliteal segments. Altogether 56 primary one-step operations were performed. The initial success was achieved in 53 (94.6%) cases. The long-term results were followed up in 43 (81.1%) patients, in the long-term period, repeated endovascular interventions were accomplished in 6 (1.8%) patients with re-stenoses of the iliac arteries. The best long-term results were obtained in the groups of patients who had undergone balloon angioplasty and stenting of the iliac arteries. PMID- 16037812 TI - Intraoperative preparation of the radial artery for coronary artery bypass grafting. AB - In coronary surgery, the problem of pharmacological prevention of vasospasm of the bypass from the radial artery (RA) remains unsolved until now. In view of this circumstance the authors propose a method for the use of aminazine (chlorpromazine), a phenothiazine series inhibitor of calmodulin, for prevention of vasospasm of auto-arterial bypasses from the RA at coronary artery bypass grafting (CABG). An analysis was made of the results of 30 CABGs using the RA conduit. The operations were performed at the Department of Cardiovascular Surgery, SI Research Institute of Cardiology, TSC, SB RAMS. The experimental data presented herein demonstrate that intraoperative treatment of the exposed RA by aminazine solution in a concentration of 10-6 M/L may prevent spasm of the bypass from the RA for a long time. Control bypassgraphy carried out over the first postoperative week has revealed that all the bypasses from the RA are functionally competent. So, the method proposed allows a reliable, effective and prevention of perioperative spasm of auto-arterial bypasses from the RA at CABG. PMID- 16037813 TI - The right aortic arch with mirror-image branching of brachiocephalic arteries and aplasia of the left brachiocephalic trunk: surgically cured rare cause of cerebrovascular insufficiency. AB - A clinical case with rare anomaly: right aortic arch with mirror-image branching of brachiocephalic arteries and aplasia (isolation) of the left brachiocephalic trunk, causing cerebrovascular insufficiency and left arm claudication is described. Despite the clear neurologic manifestations of cerebrovascular insufficiency and absence of the pulse on the left arm, the pathology was not diagnosed until 35 years of age. The anomaly was diagnosed and surgically cured in the Department of Vascular Surgery of Vilnius University Emergency Hospital. Using transsternal median thoracotomy approach, 12 mm diameter PTFE vascular graft, arising from the ascending aorta and connected to the angle, made of branching left common carotid and left subclavian arteries, was implanted into the position of missing left brachiocephalic trunk. 3 years after surgery patient feels healthy and is working. To our knowledge this is the first reported case of surgical replacement of missing left brachiocephalic trunk in the right-sided aortic arch with mirror-image branching of brachiocephalic arteries. PMID- 16037814 TI - [Varicose eczema: the etiology, pathogenesis and diagnosis. Part II]. AB - Varicose eczema is a frequently occurring complication due to chronic venous insufficiency. In view of the fact that the complicated forms of chronic venous insufficiency (classes 4-6 according to the clinical classification CEAP) are associated with the presence of dermatological lesions, the reliable diagnosis of the given pathology presents a matter of great concern. For the phlebologist end, not infrequently, for the dermatologist the diagnosis of varicose eczema may be difficult because of the diversity of its manifestations. Also, there is a number of diseases that look like varicose eczema but require a different approach to the treatment. The Russian literature practically lacks the works related to varicose eczema. The problems of the diagnosis and treatment including the organizational aspects have not been worked out. The surgeons hold that this disease is dermatological whereas the dermatologists view it as surgical. As a result, the majority of patients do not receive adequate treatment. The paper describes the phlebologist's standpoint of the problem of varicose eczema. Part II of the paper reviews the differential diagnosis of varicose eczema. PMID- 16037815 TI - [The treatment of lower limb varicosity under outpatient conditions]. AB - Presented herein is a 7-year experience with the treatment of lower limb varicosity under outpatient conditions according to the authors' technology based on three inventions. The advantages of the given treatment method as compared to the traditional surgical technology are demonstrated as are its simplicity, safety and efficiency. PMID- 16037816 TI - Interactions between NEEP21, GRIP1 and GluR2 regulate sorting and recycling of the glutamate receptor subunit GluR2. AB - Trafficking of AMPA-type glutamate receptors (AMPAR) between endosomes and the postsynaptic plasma membrane of neurons plays a central role in the control of synaptic strength associated with learning and memory. The molecular mechanisms of its regulation remain poorly understood, however. Here we show by biochemical and atomic force microscopy analyses that NEEP21, a neuronal endosomal protein necessary for receptor recycling including AMPAR, is associated with the scaffolding protein GRIP1 and the AMPAR subunit GluR2. Moreover, the interaction between NEEP21 and GRIP1 is regulated by neuronal activity. Expression of a NEEP21 fragment containing the GRIP1-binding site decreases surface GluR2 levels and delays recycling of internalized GluR2, which accumulates in early endosomes and lysosomes. Infusion of this fragment into pyramidal neurons of hippocampal slices induces inward rectification of AMPAR-mediated synaptic responses, suggesting decreased GluR2 expression at synapses. These results indicate that NEEP21-GRIP1 binding is crucial for GluR2-AMPAR sorting through endosomes and their recruitment to the plasma membrane, providing a first molecular mechanism to differentially regulate AMPAR subunit cycling in internal compartments. PMID- 16037817 TI - Nuclear export and cytoplasmic processing of precursors to the 40S ribosomal subunits in mammalian cells. AB - It is generally assumed that, in mammalian cells, preribosomal RNAs are entirely processed before nuclear exit. Here, we show that pre-40S particles exported to the cytoplasm in HeLa cells contain 18S rRNA extended at the 3' end with 20-30 nucleotides of the internal transcribed spacer 1. Maturation of this pre-18S rRNA (which we named 18S-E) involves a cytoplasmic protein, the human homolog of the yeast kinase Rio2p, and appears to be required for the translation competence of the 40S subunit. By tracking the nuclear exit of this precursor, we have identified the ribosomal protein Rps15 as a determinant of preribosomal nuclear export in human cells. Interestingly, inhibition of exportin Crm1/Xpo1 with leptomycin B strongly alters processing of the 5'-external transcribed spacer, upstream of nuclear export, and reveals a new cleavage site in this transcribed spacer. Completion of the maturation of the 18S rRNA in the cytoplasm, a feature thought to be unique to yeast, may prevent pre-40S particles from initiating translation with pre-mRNAs in eukaryotic cells. It also allows new strategies for the study of preribosomal transport in mammalian cells. PMID- 16037818 TI - Arf-dependent regulation of Pdgf signaling in perivascular cells in the developing mouse eye. AB - We have established that the Arf tumor suppressor gene regulates mural cell biology in the hyaloid vascular system (HVS) of the developing eye. In the absence of Arf, perivascular cells accumulate within the HVS and prevent its involution. We now demonstrate that mural cell accumulation evident at embryonic day (E) 13.5 in Arf(-/-) mice was driven by excess proliferation at E12.5, when Arf expression was detectable in vitreous pericyte-like cells. Their expression of Arf overlapped with Pdgf receptor beta (Pdgfrbeta), which is essential for pericyte accumulation in the mouse. In cultured cells, p19Arf decreased Pdgfrbeta and blocked Pdgf-B-driven proliferation independently of Mdm2 and p53. The presence of a normal Arf allele correlated with decreased Pdgfrbeta in the embryonic vitreous. Pdgfrbeta was required for vitreous cell accumulation in the absence of Arf. Our findings demonstrate a novel, p53- and Mdm2-independent function for p19Arf. Instead of solely sensing excessive mitogenic stimuli, developmental cues induce Arf to block Pdgfrbeta-dependent signals and prevent the accumulation of perivascular cells selectively in a vascular bed destined to regress. PMID- 16037819 TI - Deconstructing stemness. AB - Stem cells are unique in their capacity to self-renew and generate differentiated progeny to maintain tissues throughout life. A common molecular program for stem cells has remained elusive. We discuss what the molecular logic of stemness may be. We suggest that it may not be coupled to distinct cellular properties such as self-renewal or multipotency, but rather to the stable suspension at a specific developmental stage. In this view, the stem cell niche allows a cell to maintain a transcriptional accessibility enabling the generation of specific differentiated progeny. PMID- 16037821 TI - Hematopoietic stem cell gene therapy with drug resistance genes: an update. AB - Transfer of drug resistance genes into hematopoietic stem cells (HSCs) has promise for the treatment of a variety of inherited, that is, X-linked severe combined immune deficiency, adenosine deaminase deficiency, thalassemia, and acquired disorders, that is, breast cancer, lymphomas, brain tumors, and testicular cancer. Drug resistance genes are transferred into HSCs either for providing myeloprotection against chemotherapy-induced myelosuppression or for selecting HSCs that are concomitantly transduced with another gene for correction of an inherited disorder. In this review, we describe ongoing experimental approaches, observations from clinical trials, and safety concerns related to the drug resistance gene transfer. PMID- 16037820 TI - Cell cycle-dependent nuclear retention of p53 by E2F1 requires phosphorylation of p53 at Ser315. AB - We show here that the cell cycle-dependent DNA-binding and transcriptional activity of p53 correlates with E2F expression in human primary fibroblasts. E2F1 binds and stimulates DNA-binding, transactivation and apoptotic functions of p53 but not p63 and p73. E2F1 binds residues 347-370 of p53 and enhances nuclear retention of Ser315 phosphorylated p53. This regulation of p53 by E2F1 is cell cycle dependent, as the cellular distribution of Ser315 phosphorylated p53 is associated with the periodic expression of E2F and cyclin A throughout the cell cycle. This is the first demonstration that the activities of p53 are regulated during the cell cycle by E2F/p53 interactions and that phosphorylation of p53 at Ser315 is required for this regulation. PMID- 16037822 TI - Targeting proliferating tumor cells via the transcriptional control of therapeutic genes. AB - We have previously reported the construction of a cell cycle-regulated HSV-1 amplicon vector (denoted as pC8-36) that confers luciferase reporter gene activities dependent on cellular divisions. However, luciferase reporter gene is well known for its relatively high sensitivity, thus, it is crucial to evaluate the therapeutic efficacy of a transcriptional targeted vector. In this report, we have engineered the FasL and FADD genes into pC8-36 and demonstrated their efficacy for the treatment of human gliomas in vitro and in vivo. Using trypan blue dye exclusion and TUNEL assay, FasL expression mediated by pC8-36 was shown to induce a significantly higher percentage of cell death in proliferating cells than those observed in the G(1)-arrested cells. The observed cell killing effect correlated well with the level of FasL protein expression when analyzed by ELISA assay. Furthermore, the incorporation of both FasL and FADD into pC8-36 resulted in the enhancement of apoptosis in the target glioma cells both in vitro and in vivo. Targeting proliferating tumor cells via the transcriptional control of therapeutic genes could potentially improve the safety and efficacy of cancer gene therapy, and thus would allow the development of strategies for more effective anticancer therapies. PMID- 16037823 TI - Combination effect of oncolytic adenovirotherapy and TRAIL gene therapy in syngeneic murine breast cancer models. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) gene therapy and oncolytic adenovirotherapy have been investigated extensively in xenografic human tumor models established in immunocompromised nude mice. However, the effects of these therapies on syngeneic murine tumors in immunocompetent settings were not well documented. We hypothesized that TRAIL gene therapy used with an oncolytic adenovirus would overcome the weaknesses of the two therapies used individually. In this study, we evaluated the antitumor effects of an oncolytic adenovirus, Delta24, in both human and murine breast cancer cell lines. We also analyzed the effects of TRAIL gene therapy combined with oncolytic virotherapy in these cancer cells. Our results showed that Delta24 can replicate and help the E1-deleted adenovector replicate in murine cancer cells. We also found that these two therapies combined had greater antitumor activity than either one alone in both human and murine breast cancer cells lines and in the syngeneic breast cancer models established in immunocompetent mice. Moreover, Delta24 virotherapy alone and combined with TRAIL gene therapy dramatically reduced the spontaneous liver metastasis that originated in the subcutaneous 4T1 tumor established in Balb/c mice. These findings provide important considerations in the development and preclinical assessments of oncolytic virotherapy. PMID- 16037825 TI - Cryopyrin and pyrin activate caspase-1, but not NF-kappaB, via ASC oligomerization. AB - Mutations in cryopyrin and pyrin proteins are responsible for several autoinflammatory disorders in humans, suggesting that these proteins play important roles in regulating inflammation. Using a HEK293 cell-based reconstitution system that stably expresses ASC and procaspase-1 we demonstrated that neither cryopyrin nor pyrin or their corresponding disease-associated mutants could significantly activate NF-kappaB in this system. However, both cryopyrin and two disease-associated cryopyrin mutants induced ASC oligomerization and ASC-dependent caspase-1 activation, with the disease associated mutants being more potent than the wild-type (WT) cryopyrin, because of increased self-oligomerization. Contrary to the proposed anti-inflammatory activity of WT pyrin, our results demonstrated that pyrin, like cryopyrin, can also assemble an inflammasome complex with ASC and procaspase-1 leading to ASC oligomerization, caspase-1 activation and interleukin-1beta processing. Thus, we propose that pyrin could function as a proinflammatory molecule. PMID- 16037826 TI - Increased expression of matrix metalloproteinase-2 in oral cells after short-term stimulation and long-term usage of areca quid. AB - BACKGROUND AND PURPOSE: Arecoline, an areca quid (AQ) component, has been shown to inhibit the secretion and activity of matrix metalloproteinase-2 (MMP-2) in fibroblast cultures. This study assessed whether MMP-2 expression was inhibited in the saliva samples and tumor specimens of oral tumor patients with a long-term history of AQ consumption. The net effect of crude AQ extract (AQE) on MMP-2 expression by oral cells was also investigated. METHODS: Western blot analysis, zymography, and reverse transcriptase-polymerase chain reaction were used to detect MMP-2 protein and mRNA in saliva and tumor samples, as well as in the conditioned media (CM) of oral cell cultures. RESULTS: The level of MMP-2 protein was significantly higher in the saliva samples of 12 oral tumor patients who had a minimum 10-year AQ-consuming history than in those of 12 non-AQ-using healthy controls (p < 0.05). MMP-2 mRNA was expressed in 26 of 28 oral squamous cell carcinoma (OSCC) specimens. MMP-2 protein was also detectable in the tested OSCC homogenates. Short-term stimulation with 10% AQE increased the secretion of MMP-2 protein in the CM of oral epidermoid carcinoma cell Meng-1 (an OSCC cell line) and oral fibroblasts. CONCLUSIONS: MMP-2 expression is elevated rather than inhibited in most oral tumor patients with long-term AQ usage. Short-term AQE stimulation also increases the secretion of MMP-2 by oral epithelial cells and fibroblasts. Our results suggest that AQ consumption may promote oral tumor progression through the induction of MMP-2 secretion. PMID- 16037824 TI - Minimally invasive localization of oncolytic herpes simplex viral therapy of metastatic pleural cancer. AB - Herpes simplex virus-1 (HSV-1) oncolytic therapy and gene therapy are promising treatment modalities against cancer. NV1066, one such HSV-1 virus, carries a marker gene for enhanced green fluorescent protein (EGFP). The purpose of this study was to determine whether NV1066 is cytotoxic to lung cancer and whether EGFP is a detectable marker of viral infection in vitro and in vivo. We further investigated whether EGFP expression in infected cells can be used to localize the virus and to identify small metastatic tumor foci (<1 mm) in vivo by means of minimally invasive endoscopic systems equipped with fluorescent filters. In A549 human lung cancer cells, in vitro viral replication was determined by plaque assay, cell kill by LDH release assay, and EGFP expression by flow cytometry. In vivo, A549 cells were injected into the pleural cavity of athymic mice. Mice were treated with intrapleural injection of NV1066 or saline and examined for EGFP expression in tumor deposits using a stereomicroscope or a fluorescent thoracoscopic system. NV1066 replicated in, expressed EGFP in infected cells and killed tumor cells in vitro. In vivo, treatment with intrapleural NV1066 decreased pleural disease burden, as measured by chest wall nodule counts and organ weights. EGFP was easily visualized in tumor deposits, including microscopic foci, by fluorescent thoracoscopy. NV1066 has significant oncolytic activity against a human NSCLC cell line and is effective in limiting the progression of metastatic disease in an in vivo orthotopic model. By incorporating fluorescent filters into endoscopic systems, a minimally invasive means for diagnosing small metastatic pleural deposits and localization of viral therapy for thoracic malignancies may be developed using the EGFP marker gene inserted in oncolytic herpes simplex viruses. PMID- 16037828 TI - Clinical manifestations and outcomes of constrictive pericarditis in children. AB - BACKGROUND AND PURPOSE: Constrictive pericarditis (CP) is rare in children. The purpose of this study was to define the clinical manifestations and outcomes of this disease. METHODS: From January 1985 through December 2003, 8 patients with surgically confirmed CP were enrolled. Data on clinical characteristics, disease manifestations, treatment, and outcomes were collected and analyzed. RESULTS: One patient had l-transposition of the great arteries (l-TGA); the other 7 patients had no underlying cardiovascular diseases. None of the patients had direct evidence of tuberculous infection. Age at disease onset ranged from 8 months to 13 years (median, 10 years), and the interval between symptom onset and correct diagnosis ranged from 2 to 31 months (median, 12 months). Major symptoms and signs included abdominal distension, hepatomegaly, neck vein engorgement and ascites. Evidence of calcification on chest roentgenogram was found in 1 patient (12.5%). Computed tomography or magnetic resonance image studies showed pericardial thickening in 6 patients (75%). Seven patients underwent cardiac catheterization, and all hemodynamic data met the diagnostic criteria for CP. All patients with structurally normal hearts survived during a median follow-up of 31 months after pericardiectomy (range, 5 to 103 months). The patient with l-TGA died of pneumonia 1 year after pericardiectomy. CONCLUSIONS: Symptoms and signs of right heart failure were common in children with CP. The etiology of CP remains largely unknown. Patients with structurally normal hearts have good outcomes after pericardiectomy. PMID- 16037827 TI - Intussusception in Taiwanese children: analysis of incidence, length of hospitalization and hospital costs in different age groups. AB - BACKGROUND AND PURPOSE: This study investigated the characteristics of intussusception in Taiwanese children of different age groups, including the incidence, length of hospitalization and hospital costs. METHODS: Children with a diagnosis of intussusception who were hospitalized from 1999 through 2001 were identified from a nationwide health insurance claims database. The incidence of intussusception was calculated by age, gender, and season. Length of hospitalization and hospital costs were also analyzed. RESULTS: A total of 6988 cases of intussusception were identified in Taiwan from 1999 to 2001. Among them, 4859 cases occurred in children below 15 years of age. The average incidence among children below age 15 years was 34.5 per 100,000, with a peak incidence of 118.8 per 100,000 observed among children younger than 24 months old. The highest incidence of intussusception in Taiwanese children occurred between 12 and 24 months of age. According to the data for patients below 15 years of age hospitalized for intussusception in year 2000, males were more likely to be affected than females (61.3% vs 38.7%). Intussusception-related hospitalizations were rare in infants in the first few months of life, increased in those 6 to 12 months old, and peaked among children 1 to 3 years old. Among the 952 patients with intussusception admitted to hospitals in 2000, 297 (31.2%) received surgery, incurring higher median medical costs (New Taiwan Dollars [NT dollars] 42,265 or US dollars 1234) and longer median hospital stay (6.2 days) than the 655 patients who did not require surgery (NT dollars 6290 or US dollars 185 for hospitalization of 2.4 days). CONCLUSIONS: The study found that the incidence of intussusception peaked in the second year of life in Taiwanese children. There was also a male predominance and lack of seasonal variation in incidence. PMID- 16037829 TI - Cardiac catheterization in low birth weight infants. AB - BACKGROUND AND PURPOSE: The increased survival of low birth weight infants means that more of these infants may be candidates for cardiac catheterization. There is a lack of data from Taiwan on the results of cardiac catheterization in these infants. This study investigated the outcome of cardiac catheterization in infants weighing less than 2500 g in a single hospital over an 11-year period. METHODS: The records of 41 infants (17 males and 24 females) weighing less than 2500 g at the time of cardiac catheterization from August 1993 to July 2004 were reviewed. Morbidity and mortality were compared between diagnostic and intervention groups. RESULTS: Interventional procedures were performed in 14 and diagnostic catheterizations in 27 infants. These interventional procedures included 5 balloon valvuloplasties for pulmonary stenosis and 1 for aortic stenosis, 1 balloon angioplasty for critical coarctation, 1 aortic valvuloplasty, 1 dilatation for coarctation plus aortic valvuloplasty, 1 temporary pacemaker implantation, and 5 balloon atrioseptostomies. The median age at catheterization was 5 days and the median weight was 2.35 kg. All diagnostic and interventional procedures were successfully performed without mortality. Complications included 2 cases each of arrhythmia, bleeding, apnea, and thromboembolism in the diagnostic group and, in the intervention group, 2 cases each of supraventricular tachycardia and bleeding, and 1 case of linear dissection of a coarctation. CONCLUSIONS: None of the 41 catheterized low birth weight infants died. Furthermore, the complication rate in the intervention group was no higher than in the diagnostic group. We conclude that cardiac catheterization, even with an interventional procedure, can be performed safely in low birth weight infants. PMID- 16037831 TI - Isolation of the left subclavian artery in a child with tetralogy of Fallot and right aortic arch. AB - Isolation of the subclavian artery is a very rare vascular anomaly in which the subclavian artery does not communicate with the aortic arch but instead is connected to the pulmonary artery by the ductus arteriosus. The subclavian steal phenomenon from the vertebrobasilar system into the pulmonary and subclavian arteries is usually persistent. We report an 18-month-old boy with an isolated left subclavian artery associated with tetralogy of Fallot and right aortic arch. Subclavian steal phenomenon in this patient resulted in symptoms of diminished pulse and blood pressure of the left arm. He underwent total correction of tetralogy of Fallot and ligation of the ductus arteriosus without reimplantation of the left subclavian artery. There was no substantial blood pressure difference between the upper limbs after the operation. In a patient with right aortic arch and diminished blood pressure or pulse in the left arm, isolation of the left subclavian artery should be included in the differential diagnosis. PMID- 16037830 TI - Prevalence of anisometropia in Taiwanese schoolchildren. AB - BACKGROUND AND PURPOSE: This study investigated the prevalence and distribution of anisometropia in Taiwanese schoolchildren using nationwide data from refractive surveys performed in 1995 and 2000. METHODS: Complete survey data was obtained for 11,175 students in 1995 and 10,878 students in 2000. The refractive status of each student was measured using an autorefractometer under cycloplegia and rechecked with retinoscopy. The difference in refractive status between each participant's eyes was determined. Chi-squared statistic was used to assess the difference between the 2 surveys. Multiple linear regression was used to determine the trend and effects of covariates. RESULTS: Most of the schoolchildren (77.6% in 1995, 71.9% in 2000) were not anisometropic. Most of the anisometropic differences were in the range 0.5 to 1.0 D (14.1% in 1995 vs 17.9% in 2000). About 6% of schoolchildren in 1995 and 7.0% in 2000 had anisometropic differences in the range from -1.0 to -2.0 D. Fewer than 4% of students had a level of anisometropia greater than 2.0 D (2.7% vs 3.2%, respectively). The prevalence of anisometropia and the extent of anisometropic difference both increased with age and with maximal myopic refraction (both p < 0.0001). Both the prevalence and extent of anisometropia showed significant differences between the 2 surveys (both p < 0.0001). CONCLUSIONS: Most of the Taiwanese schoolchildren surveyed were non-anisometropic. The prevalence and amount of anisometropia were significantly increased from 1995 to 2000. The mechanisms responsible for these increases have not been determined, but may be related to increase of myopic refraction. PMID- 16037832 TI - Recurrent acalculous cholecystitis and sclerosing cholangitis in a patient with X linked hyper-immunoglobulin M syndrome. AB - X-linked hyper-immunoglobulin M (IgM) syndrome (XHIGM) is a rare genetic primary immunodeficiency disease caused by mutations of the CD40 ligand (CD40L) gene with normal or elevated levels of IgM and markedly decreased serum IgG, IgA, and IgE. Liver disease may occur as a clinical manifestation in XHIGM. This complication appears to increase with age. We report an 18-year-old male patient who had recurrent episodes of acalculous cholecystitis (AC) and sclerosing cholangitis (SC). The diagnosis of XHIGM was confirmed by the finding of CD40L expression < 1% of normal and a tyrosine 169 asparaginase (t526a) mutation in exon 5 (the tumor necrosis factor domain) of the CD40L gene. The patient had direct hyperbilirubinemia (direct bilirubin 5.5 mg/dL, total bilirubin 8.7 mg/dL), cholestasis (alkaline phosphatase 1133 U/L, gamma-glutamyl transferase 1019 U/L) and elevated transaminases (aspartate aminotransferase 70 U/L, alanine aminotransferase 101 U/L). Findings on abdominal ultrasound and abdominal computed tomography were compatible with AC. After the fourth episode of cholecystitis, cholecystectomy and liver biopsy were performed. Operative cholangiography revealed poor opacification of the hepatic duct and proximal common bile duct; the upstream intrahepatic bile ducts were not visualized. The biopsy specimen showed marked fibrosis of the portal areas. Enterococcus species was cultured from the bile. Children or adolescents with recurrent AC and SC should be evaluated for an underlying immunodeficiency syndrome such as XHIGM. PMID- 16037833 TI - Dental anomalies in two patients with incontinentia pigmenti. AB - Incontinentia pigmenti (IP) is a rare X-linked dominant inherited disorder which has a variety of ectodermal aberrations. Skin hyperpigmentation is the most characteristic feature of IP. However, extracutaneous anomalies involving dentition, hair, eyes, and central nervous system are also found. The dental anomalies reported include peg-shaped or malformed teeth, hypodontia, delayed eruption, and impacted tooth. This report describes the dental anomalies in 2 IP patients who had the characteristic features of skin hyperpigmentation. One was a 13-year-old girl who had slender cone-shaped permanent anterior teeth, hypodontia, and delayed eruption of teeth which are characteristic dental anomalies in an IP patient. The other was a 10-year-old girl who only had 2 tulip shaped maxillary permanent central incisors with shorter tapering roots but no congenital missing teeth or delayed eruption of teeth. Our findings suggest that IP may present a broad variation of dental anomalies individually. However, the characteristic finding of permanent anterior teeth with a longer crown and a shorter root found in both of our IP patients may be worthy of consideration in the differential diagnosis of IP. PMID- 16037834 TI - Acute pancreatitis combined with acute Budd-Chiari syndrome as the initial manifestation of small cell lung cancer. AB - Tumor metastasis to the pancreas is a rare but recognized cause of acute pancreatitis. Autopsy series have reported a 24-40% of pancreatic involvement in small cell lung cancer. However, only a very few cases of tumor-induced acute pancreatitis have been described. Budd-Chiari syndrome complicating lung cancer is a rarely reported condition. We report a 68-year-old woman with extensive small cell lung cancer with the unusual initial presentation of both acute pancreatitis and acute Budd-Chiari syndrome. This patient suffered from progressive epigastralgia for 3 weeks. Severe epigastralgia with radiation to back and progressive jaundice developed 2 days prior to admission. After admission, the liver enlarged rapidly and the ascites increased markedly. Chest roentgenogram showed a mass lesion over the left lower lung field. Poorly differentiated carcinoma cells were found in ascites and bone marrow. The patient died on the ninth day of hospitalization before chemotherapy was initiated. Prompt diagnosis of extensive-stage small cell lung cancer may allow early chemotherapy treatment which favorably influences recovery when the pancreatitis is mild. Although prolonged survival might have been expected had this patient recovered from pancreatitis and received chemotherapy, diagnosis was delayed due to difficulty in immunohistochemical diagnosis of the tumor and the unusual clinical presentation. The use of stains employing antibodies against neurofilament and neuron-specific enolase cell antigens is important for early diagnosis of poorly differentiated metastatic tumor cells. PMID- 16037835 TI - Metachronous adenocarcinoma occurring at an esophageal colon graft. AB - Subtotal esophagectomy and reconstruction with colonic interposition grafting is the usual surgical treatment for benign esophageal stricture and malignant esophageal cancer. While short-term complications are frequently reported, long term complications are rare. Herein, we report a 57-year-old male, with a previous caustic injury to the esophagus who underwent subtotal esophagectomy and right colon grafting at age 18. He underwent left hemicolectomy for obstructive colon cancer at the splenic flexure at age 55, followed by resection of the interposed colon graft due to adenocarcinoma at age 57. PMID- 16037836 TI - Use of magnetic resonance cholangiopancreatography to diagnose neonatal congenital choledochal cyst. AB - Antenatal identification of choledochal cysts has become increasingly common with advances in prenatal ultrasonography. However, the antenatal diagnosis needs to be confirmed postnatally and a preoperative radiologic evaluation of the cyst prior to complete resection is required. There have been few reports of the application of magnetic resonance cholangiopancreatography in neonates, or in neonates with large choledochal cysts. We describe a neonate with a progressively growing giant choledochal cyst, which was initially detected at 28 weeks' gestation. After delivery, the baby did not feed well and suffered from jaundice and frequent postprandial bilious vomiting. At the age of 11 days, magnetic resonance cholangiopancreatography was used to confirm the prenatal diagnosis and provide a thorough preoperative evaluation. He underwent early resection of the cyst at 15 days of age and recovered uneventfully. Magnetic resonance cholangiopancreatography is an alternative diagnostic method for confirming the antenatal diagnosis of large choledochal cysts in neonates. It may also provide useful diagnostic information for preoperative evaluation. PMID- 16037837 TI - Prevalence and associated factors of nocturia among adult residents of the Matsu area of Taiwan. AB - The aims of this study were to investigate the prevalence of nocturia among adults residing in the Matsu area of Taiwan, and to examine the association of this condition with sociodemographic and health characteristics. Between October 2002 and February 2003, all adults older than 30 years residing in the Matsu area were invited to participate in a health awareness campaign. A total of 1706 adults aged 30 to 91 years completed the study survey. Among them, 16.8% reported nocturia with > or = 2 voids per night. The prevalence of nocturia increased with age from 8.4% in the 30-39 years age group to 30.9% in the > or = 70 years age group (p < 0.001). No significant difference of nocturia prevalence was found between men and women. In addition to age, diabetes (odds ratio, 2.5; 95% confidence interval, 1.4-4.2) was also independently associated with nocturia. This study suggests the importance of nocturia as a health problem and the need for greater awareness and treatment. PMID- 16037838 TI - Matched-unrelated-donor bone marrow transplantation for children with leukemia. AB - BACKGROUND AND PURPOSE: This report describes the results of matched-unrelated donor transplant for leukemia or myelodysplasia in the first 23 recipient children at a single medical center in Taiwan. METHODS: Between August 1994 and February 2003, 23 consecutive children with leukemia or myelodysplasia underwent matched-unrelated-donor bone marrow transplantation (BMT). The preparative regimen consisted of fractionated total body irradiation and cyclophosphamide in 6 patients; busulfan in combination with etoposide and cyclophosphamide in 4 patients who received cranial irradiation before transplantation; and busulfan and cyclophosphamide in 13 patients. RESULTS: Engraftment was achieved in 91.3% of cases. Acute graft-versus-host disease (GVHD) occurred in 18 of 21 patients who engrafted (85.7%). Event-free survival for all patients was 24.46 +/- 9.24%. The 12 children with standard-risk disease had better event-free survival than the 11 children with high-risk disease (46.88 +/- 15.03% vs 0%, p < or = 0.001). CONCLUSIONS: The major obstacles to successful matched-unrelated-donor BMT are acute GVHD, relapse and infection. Early transplantation and patient selection, prophylactic and therapeutic maneuvers for GVHD, as well as appropriate donor selection and virus prophylaxis may improve the results. PMID- 16037839 TI - Interscience Conference on Antimicrobial Agents and Chemotherapy--40th meeting. Faropenem: a new antimicrobial agent. 17-20 September 2000, Toronto, Canada. PMID- 16037840 TI - Moxifloxacin: a new antimicrobial agent. PMID- 16037841 TI - BMS-284756 and ABT-773: new antibacterial agents. PMID- 16037842 TI - Highlights of new antimicrobial agents. PMID- 16037843 TI - Antifungal agents and efflux pump inhibitors. PMID- 16037845 TI - Poster and slide session I. PMID- 16037844 TI - Selected symposia. PMID- 16037846 TI - Poster and slide session II. PMID- 16037847 TI - Antibiotics and ribosomes: interaction and resistance. AB - This session was convened by Dr Joyce Sutcliffe (Pfizer Central Research, Groton, CT, USA) and Dr R Leclerq (Hopital Cote de Nacre, Caen, France), with the assistance of the Committee Member Advisor, Dr Patrice Courvalin (Institut Pasteur, Paris, France). It included several presentations in which recent advances on understanding interactions of antibiotics with the ribosome, and mechanisms of action of such drugs, were discussed. The majority of speakers focused on antibiotic interaction with the large ribosomal subunit. Exciting new developments in the field of ribosomal research were highlighted by publication, just several weeks before the conference, of high-resolution crystallographic structures of the large and small ribosomal subunits. With this in mind, several speakers discussed biochemical and genetic data on antibiotic-ribosome interactions within the context of newly available structural information. PMID- 16037848 TI - Global Resistance Day. PMID- 16037850 TI - Poster sessions. PMID- 16037849 TI - Inflammation Research Association--Tenth National Meeting Symposia. 24-28 September 2000, Hot Springs, VA, USA. PMID- 16037852 TI - Clinical trials of new drugs. PMID- 16037853 TI - Leukemia 2000: towards the cure. 7-9 September 2000, Houston, TX, USA. PMID- 16037855 TI - Postmenopausal osteoporosis. 22-26 September 2000, Toronto, Canada. PMID- 16037854 TI - American Society for Bone and Mineral Research--22nd Annual Meeting. PMID- 16037856 TI - Nucleosides, nucleotides and their biological applications--XIV International Roundtable. Recent Advances in Nucleosides Chemistry and Chemotherapy. 10-14 September 2000, San Francisco, CA, USA. AB - This satellite symposium honoring Dr Jack J Fox, provided a forum for reviewing recent advances in nucleoside research. The topics covered were: novel chemistries, applications to current problems in biology, antiviral and antitumor agents, and aspects of enzymology and mechanism of action of important nucleoside analogs. PMID- 16037857 TI - American College of Clinical Pharmacology--29th Annual Meeting. 17-19 September 2000, Chicago, IL, USA. PMID- 16037858 TI - Medicinal chemistry--16th international symposium. 18-22 September 2000, Bologna, Italy. AB - This meeting was part of a biennial series focusing upon topics at the forefront of medicinal chemistry. There was no single therapeutic focus for the meeting, instead, the lectures, arranged with main lectures taking place in three parallel sessions, were spread over a number of themes relevant to drug discovery. These are highlighted in order of presentation. PMID- 16037859 TI - Photodynamic therapy comes of age. AB - The concept underlying photodynamic therapy (PDT) is the use of light-absorbing molecules which, when delivered to target cells, and activated by irradiation with light of the appropriate wavelength, produce reactive oxygen species that cause cell death by apoptosis or necrosis. Classically, photodynamic agents have been macrocycles and, as such, application is limited to topical and intravenous administration. In the latter case, reliance has been placed on the characteristic behavior of the photodynamic agents in showing some degree of selectivity for concentrating in the target to minimize non-specific damage to the host tissue. The parameters open to development of improved drugs are: (i) the design of the photodynamic agent molecule as a means of determining the wavelength of light for activation, and for influencing physicochemical characteristics and the pharmacokinetic behavior of the drug; (ii) the delay between administration and activation; (iii) the nature of the activating light source; and, (iv) the duration and intensity of the activating light. Obviously, PDT is attractive for treating disease states in which natural apoptotic mechanisms are compromised, specifically for cancerous states and in cases of uncontrolled cell proliferation. PDT also has immunomodulatory sequelae, including triggering of T-cell mediated activity against residual cancerous cells. The use of PDT is being extended to diverse, related immune and proliferative disease states, and to the inactivation of bacteria and viruses. Increasingly, attention is being given to improving treatment by targeting conjugates and local delivery strategies, as well as by the design of photodynamic agents with closely defined photophysical and physicochemical properties. Progress is being made in challenging indications, such as the treatment of solid and pigmented tumors. Alternative technologies not involving light activation are available for some molecules which may also be used with light activation. Some ex vivo techniques and medical devices have been reported. PMID- 16037860 TI - Factor Xa inhibitors--a review of the recent patent literature. AB - This review covers the patent literature and related scientific reports in the field of factor Xa inhibitors published between January 1999 and June 2000. During this time, the amount of scientific information as well as the number of newly published patent applications has continuously increased. It is the aim of this review to give an overview of the different structural types of factor Xa inhibitors, to clarify the relationship between newly claimed and previously known inhibitors, and assess the clinical potential of the different factor Xa inhibitors. PMID- 16037861 TI - Decitabine (SuperGen). AB - Decitabine, a potent DNA methyltransferase inhibitor, which was originally under development by Pharmachemie, is being developed by SuperGen. Pharmachemie had been studying decitabine in phase II clinical trials for several leukemia indications in Europe and the US. Preliminary results indicated that the compound was active in the treatment of myelodysplasia, relapsed leukemia, acute myeloid leukemia and postallogeneic progenitor cell transplant relapse. The compound is in phase II clinical trials with phase III trials scheduled to begin shortly. Decitabine has been used to treat myelodysplastic syndrome in a total of 125 patients, with an overall response rate of 49%. In a study using decitabine to treat chronic myelogenous leukemia in 81 patients, a response rate of 62% among patients in chronic phase of the disease was achieved. In a phase I/II trial designed to establish safety and efficacy in the treatment of sickle cell anemias treatment with decitabine generated a response in 100% of the patients tested: a total of eight patients were enrolled, each experienced elevated levels of fetal hemoglobin. Side effects were minimal and the drug was well tolerated. Plans for additional clinical studies of decitabine as a treatment for sickle cell anemia are underway. A phase II trial using a low dose of decitabine in patients with myelodysplastic syndrome has been completed. Of 66 patients entered, 62 were evaluable. The response rate was 48%, with a median response duration of 40 weeks. The mean survival from the start of therapy was 13 months. In a study with 37 CML patients, a 25% overall response rate was seen in those patients in the blastic phase of the disease, and a 52% response rate was observed in the accelerated phase patients. The most significant side effect was prolonged myelosuppression. The drug suppresses cellular growth in seven human tumor cell lines, possibly by reactivation of certain growth suppressor genes. PMID- 16037862 TI - Azimilide (Procter & Gamble). AB - Azimilide is a class III anti-arrhythmic agent under development by Procter & Gamble. In December 1998, the company submitted an NDA with the FDA seeking an indication for the maintenance of sinus heart rhythm in patients with various forms of supraventricular arrhythmia. By November 1997, the drug was being evaluated in clinical trials for its use in patients with implantable cardioverter defibrillators (ICDs). The study was to examine its ability to reduce the frequency and severity of ICD electrical discharges. As of August 1999, azimilide was in phase I development by Tanabe Seiyaku in Japan, for the potential treatment of arrhythmia. Results of a pivotal clinical trial, presented at the annual meeting of the Society for Neuroscience in November 1998, demonstrated azimilide to prolong the arrythmia-free period in patients suffering from atrial fibrillation. Phase III clinical trials so far have involved patients with supraventricular and ventricular arrhythmias. Trial results from the ASAP (Azimilide Supraventricular Arrhythmia Program Trial), showed that azimilide significantly prolonged the arrythmia-free period in patients suffering from atrial fibrillation. There were three deaths in the azimilide group compared to one in the placebo group. The safety and efficacy of azimilide in 6000 patients who have suffered a heart attack is currently under study in the phase III ALIVE (azimilide post-infarct survival evaluation) multicenter trial, which commenced in centers in the US and Europe in October 1997. PMID- 16037863 TI - Victory by stages: the "tour de force" to early correction in congenital heart surgery. PMID- 16037864 TI - [Transplantation of internal thoracic artery, historical perspective]. PMID- 16037865 TI - Impact of implantation technique on hemodynamic results of the Pericarbon Freedom stentless valve. AB - BACKGROUND: The Pericarbon Freedom stentless valve has shown excellent hemodynamic results in the midterm course. However, there is no information as to whether a continuous or interrupted suture technique at the inflow site has an impact on postoperative hemodynamics. METHODS: 139 patients were enrolled in a non-randomized, prospective matched trial. An interrupted suture line technique was used in 68 patients and a continuous suture line technique was used in 71 at the inflow site. Isolated valve replacement was performed in 70.4% of the continuous and 67.6% of the interrupted suture group. Pre- and postoperative hemodynamics and one-year follow-up were obtained by echocardiography and expressed as mean and peak gradients and grade of regurgitation. RESULTS: No significant difference between continuous and interrupted suture techniques were noted with respect to mean (11.8 +/- 6.3 vs. 12.5 +/- 6.2 mm Hg, p = 0.251) and peak gradients (21.0 +/- 9.6 vs. 22.0 +/- 10.9 mm Hg, p = 0.292) as well as to the degree of regurgitation. Bypass and cross-clamping times decreased by 22.4 and 20.6 minutes, respectively, with the use of the continuous suture technique. One year follow-up showed a further, significant decrease of mean and peak gradients. CONCLUSIONS: The Pericarbon Freedom stentless valve appears to offer excellent postoperative performance. The suture line technique at the inflow site does not result in any hemodynamic differences. PMID- 16037866 TI - Intraoperative bypass flow measurement reduces the incidence of postoperative ventricular fibrillation and myocardial markers after coronary revascularisation. AB - OBJECTIVE: Sudden ventricular fibrillation (VF) and myocardial infarction (MI) are life-threatening complications after coronary artery bypass grafting (CABG). We prospectively analysed the impact of intraoperative bypass flow measurement with the transit time flow Doppler method (TTFD) on the incidence and outcome of postoperative VF and MI. METHODS: In 1995 a standardized algorithm for the treatment of postoperative VF was introduced in our institution. The rate of postoperative VF was therefore exactly registered. In 1998 the TTFD method was implemented as a standard in all CABG cases. Whenever insufficient bypass graft flow was detected, anastomoses were redone and technical problems affecting the grafts were excluded. The incidence of postoperative VF and CK/CK-MB fraction was observed prospectively and the new data was compared to the data from 1995 to 1998. RESULTS: From 1/95 to 7/98 a total of 4321 patients (group A) were operated on with isolated CABG procedures using extracorporeal circulation. In the period from 8/98 to 10/02 a total of 3421 patients (group B) was operated on with isolated CABG procedures under the same conditions, except that the TTFD method was used in every case. The treatment of VF was standardised in both groups according to the algorithm. The most striking effect was the significant reduction of VF from 0.66% to 0.44% when TTFD was introduced and the steep decrease in mortality from 30% to 12.2% in patients with VF when the algorithm and TTFD were routinely applied. Furthermore the rate of insufficient bypass flow detected by angiography was reduced by 66%. CONCLUSION: Routinely the use of TTFD significantly reduced the incidence of postoperative VF, postoperative CK/CK-MB fraction, and angiographically detected bypass malfunction. A simultaneously implemented algorithm reduced the mortality with VF after CABG. The consequent use of TTFD intraoperatively reduced the incidence of postoperative anastomosis and technically related complications of bypass surgery and led to a significant reduction of postoperative mortality in CABG procedures. PMID- 16037867 TI - Replacement of a severe chronic post-traumatic aneurysm of the ascending aorta with aortic valve conduit--reconstruction of the anterior mitral valve ring and implantation of A-V sequential/biventricular pacemaker. AB - We present the case of a 23-year-old African professional footballer who was admitted on April 1, 1999 to the Cardiology Department of the University Hospital in Magdeburg, on an emergency basis, from a regional lung clinic. According to the history, he was involved in a collision with an opposing player during a football match in his country (in Africa). He lost consciousness for a short time, but continued playing to the end of the match. About two months later he was invited by a German football club for a check-up, with the view to ultimately playing for the club. The team did not find him physically fit enough to play professional football, so he decided to go to Paris by bus on March 31, 1999. During the journey he suddenly became cardio-pulmonary decompensated and had to undergo cardio-pulmonary resuscitation (CPR). He was intubated and placed on a respirator and immediately transferred to a nearby lung clinic. From the lung clinic he was transferred to the Intensive Care Unit of the Cardiology Department of the Magdeburg University Hospital, on April 1, 1999 as an emergency case. He was intensively treated with catecholamines, intravenous ACE inhibitors and diuretics. His clinical condition did not improve appreciably. His chest X-ray showed extreme dilatation of the right and left heart as well as extreme pulmonary congestion. PMID- 16037868 TI - Hemodynamic performance and incidence of patient-prosthesis mismatch of the complete supraannular perimount magna bioprosthesis in the aortic position. AB - BACKGROUND: Complete supraannular placement and smaller stent design allow the implantation of a Perimount Magna bioprosthesis with a larger inner diameter than that of a standard Perimount. This study compares the hemodynamic performance and the incidence of patient-prosthesis mismatch (PPM) of both prostheses. METHODS: 128 patients underwent aortic valve replacement, receiving either a Magna (n = 57) or a standard (n = 71) prosthesis. Inner aortic annulus diameter was measured intraoperatively by a hegar dilator to match echocardiographically obtained results to the annulus diameter instead of matching them to labelled valve size. RESULTS: The Magna was significantly superior with respect to mean pressure gradient and effective orifice area in patients with an annulus diameter of 22-23 mm. In patients with an annulus diameter < 22 mm or > 23 mm, there was a non significant trend towards superior hemodynamics in the Magna group. Severe PPM (effective orifice area index < or = 0.65 cm (2)/m (2)) was present in 11.1% (Magna) vs. 42.1% (Standard) of patients with an annulus diameter < 22 mm; in 0% (Magna) vs. 13.8% (Standard) with an annulus diameter of 22-23 mm; no PPM was seen in patients with annulus diameter > 23 mm in both groups. CONCLUSIONS: The Perimount Magna had a significantly reduced incidence of patient-prosthesis mismatch and superior hemodynamics compared to the standard Perimount. PMID- 16037869 TI - VATS with a supraclavicular window for huge substernal goiter: an alternative technique for preventing recurrent laryngeal nerve injury. AB - A careless attempt to remove a huge substernal goiter using the cervical approach can lead to recurrent laryngeal nerve injury, which has been consistently reported after the surgery. We present an alternative and less invasive technique combining video-assisted thoracoscopic surgery (VATS) with a supraclavicular approach. This technique seems to offer improved exposure and reliable control of the neuro-vascular structures in the anterior mediastinum when resecting a huge substernal goiter that may prevent nerve injury. PMID- 16037870 TI - Surgical resection of secondary chest wall tumors. AB - OBJECTIVE: The objective of this study was to evaluate which factors influence survival following surgical resection of secondary tumors of the chest wall (non bronchial carcinoma). METHODS: Between 1990 and 2001, 69 patients (23 men, 46 women) underwent chest wall resection with curative intent. All of the patients were retrospectively analyzed for sex and age, presenting symptoms, tumor location, disease-free interval, histology, radiation therapy or chemotherapy, surgical techniques and extent of resection, 30-day mortality and long-term survival. RESULTS: The most common tumors were isolated locally recurrent breast cancer (n = 33) and renal cell carcinoma (n = 17). Resection of chest wall tumors in all of the other patients revealed a kaleidoscope of different pathologies (n = 19). Overall 5-year survival was 38 %. In patients with isolated recurrence of breast cancer and in patients with chest wall metastases of renal cell cancer, the median survival was 40.6 months and 53.7 months, respectively. A disease-free interval of more than 24 months and no systemic chemotherapy after mastectomy were parameters for a favorable prognosis in patients with breast cancer. CONCLUSIONS: We conclude that chest wall resection of secondary chest wall tumors is a safe and effective treatment as part of a multidisciplinary approach. The role of surgery will continue to evolve as improvements in systemic treatment occur. PMID- 16037871 TI - Spontaneous hemopneumothorax: is conservative treatment enough? AB - PURPOSE: Spontaneous hemopneumothorax is a rare disorder, occurring in 1% to 12% of patients with spontaneous pneumothorax. The present review was undertaken to emphasize the potential life-threatening condition of spontaneous hemopneumothorax and reassess the benefit of conservative treatment with chest tube drainage. MATERIAL AND METHODS: From 1997 to 2002, 291 cases of spontaneous pneumothorax were treated in our department. Of these, 9 (3.09%) developed hemopneumothorax (> 400 ml). The clinical features of these patients and the results of conservative and surgical management were retrospectively reviewed. RESULTS: Seven patients were treated conservatively and two required VATS and thoracotomy because of worsening clinical condition. The amount of aspirated blood ranged from 400 to 3700 ml (mean, 1533 ml). Six patients received a homologous blood transfusion. CONCLUSIONS: In conclusion, hemopneumothorax is a serious condition complicating spontaneous pneumothorax. Conservative treatment is adequate in most cases and should be performed if bleeding persists for less than 24 hours after chest tube placement. PMID- 16037872 TI - Mediastinal cystic lymphangioma: is resection always necessary? AB - Mediastinal cystic lymphangiomas are rare tumours. Usually asymptomatic, they can be complicated by a chylopericardium or/and chylothorax. We report a case of a left lesion infiltrating through the supra-aortic vessels complicated with a iatrogenic left chylothorax managed by bilateral video-assisted thoracoscopy at the same time. First, on the right, the chylous inflow was stopped by suturing the thoracic duct; then, on the left, a fenestration was done to confirm the diagnosis and treat the lesion while preserving the adjacent structures. PMID- 16037873 TI - Lung function in patients operated for chronic pleural empyema. AB - OBJECTIVES: To evaluate lung function after lung decortication in patients operated for chronic pleural empyema. METHODS AND MATERIALS: To determine the lung function after lung decortication in 20 men (71.4%) and 8 women (28.6%) a prospective evaluation of blood gases, spirometry, lung perfusion, ventilation, and alveolar permeability were performed 28 weeks (15-60 weeks) after the operation. RESULTS: Median lung perfusion on the affected side was 45.2% (26.1 55.3%) in 13 right and 36% (13.8-47.2%) in 15 left empyemas, and was within normal limits in 4/28 cases. The mean lung ventilation was 44.4% (21.5-54%) and 37% (18.9-50.6%) in patients affected on the right or left side, respectively. Ventilation perfusion inequality occurred in 22 patients (78.6%). Dynamic ventilation scintigraphy revealed impairment of epithelial integrity in 10 cases (35.7%). Individual analysis revealed that 15 patients (53.6%) showed a severe reduction in one of the examinations and 8 (28.6%) in more than one. CONCLUSIONS: Although the separate analysis of the different functional parameters revealed a mild reduction of lung function after decortication, in the majority of patients the impairment was more severe due to multilevel functional lung damage. PMID- 16037874 TI - Ruptured hydatid cyst in the right thorax: differential diagnosis to pleural empyema. AB - We present a case of a hydatid cyst (Echinococcus granulosus) rupturing into the right pleural cavity. Clinical and radiological findings were consistent with a pleural effusion caused by pneumonia and led primarily to a wrong diagnosis. Pleural fluid analysis was suspicious for parapneumonic pleural empyema. After thoracoscopy led to the correct diagnosis a combined surgical and chemotherapy approach achieved complete remission. PMID- 16037875 TI - Successful extracorporeal lung assistance for overwhelming pneumonia in a patient with undiagnosed full blown aids--a controversial therapy in HIV-patients. AB - A case is described of overwhelming pneumonia in a patient with a history of coughing since three months. Because of hypoxia and hypercapnia that could not be managed by optimal mechanical ventilation, the patient required urgent extracorporeal lung assistance (ECLA, also known as ECMO). Afterwards the diagnosis of full blown AIDS was made. Appropriate antiviral, antibiotic and antimycotic therapy was successfully established. The patient was weaned from ECLA 4 days later, and discharged from hospital after two months. PMID- 16037876 TI - Pulmonary embolism after cesarian section due to heparin-induced thrombocytopenia despite normal platelet count. AB - Heparin-induced thrombocytopenia (HIT) type II is typically characterized by a decrease in platelet count to values between 20 and 120 x 10 (9)/L or a platelet count fall of greater than 50%. We report on a patient who developed a HIT syndrome, thrombosis of the vena cava, and fulminant pulmonary embolism during heparin treatment after cesarean section, without a significant decrease in platelet count. Lepirudin anticoagulation and ecarin clotting time (ECT) monitoring were used successfully during cardiopulmonary bypass. PMID- 16037877 TI - Repair of recurrent coarctation using an ascending aortic autograft. AB - Results of aortic arch repair for interrupted aortic arch or aortic coarctation have considerably improved. However, re-stenosis or aneurysm formation is a common complication requiring complex re-interventions or even extra-anatomic bypass grafting. In two patients with recurrent coarctation, the use of cardiopulmonary bypass was mandatory, in one due to the concomitant repair of the intra-cardiac defect, in the other due to the small aortic arch, the long segment aortic coarctation and the small diameter of the supra-aortic vessels. In both patients a segment of the ascending aorta was interposed between the distal aortic arch and the proximal descending aorta with uneventful postoperative courses and freedom from pathological findings at 1 year and 6 months follow-up. In patients undergoing complex congenital heart surgery involving the ascending aorta, a segment of the autologous ascending aorta may be used to repair recurrent isthmic stenosis, avoiding the use of any foreign material. PMID- 16037878 TI - A giant rhabdomyoma with left ventricular inflow occlusion and univentricular physiology. AB - During a routine prenatal ultrasound a huge tumor was diagnosed located in the left ventricle and left atrium with complete obstruction of the left ventricular inflow tract. On the first day of life, urgent surgical intervention was necessary because of progressive heart failure with univentricular physiology. Intraoperatively, the tumor mass was found to originate from the left ventricular wall, reaching into the left atrium. Histological examination revealed a rhabdomyoma. PMID- 16037879 TI - Hemoglobin and hematocrit values after saline infusion and tourniquet. AB - This study attempted to contribute to standardization of blood testing in sport, and to investigate the effect of artificial dilution with saline. In 10 healthy, physically active males and 3 healthy physically active females hemoglobin (Hb), hematocrit (Ht), and % reticulocytes (%retics) were measured at different time points to look for possible fluctuations during day time, while the subjects had regular coffee breaks and lunch. In 7 of the subjects in a separate experiment 500 ml of saline were infused around 8 am and Hb, Ht, and %retics were measured before and every hour thereafter until 7 hours after infusion. In addition Ht was measured on a hematological analyzer as well as with a centrifuge. In a separate experiment the effect of tourniquet duration on Hb and Ht was studied in 9 of the subjects. The results show that Hb, Ht, and %retics are stable from 8 am to 4 pm, but that infusion of 500 ml of saline induces an acute decrease in Hb and Ht within one hour (Hb decreased from 15.2+/-0.9 g/dl to 14.5+/-1.0 g/dl, and Ht from 45.6+/-2.8 % to 44.0+/-2.5 %). The decline in Hb and Ht was maintained during the 7-hour observation period. Ht values of the same samples measured with a hematological analyzer and a centrifuge were not different. Application of the tourniquet did significantly affect Hb and Ht values only from two minutes, and thereafter Hb and Ht remained stable during the rest of the 5-minute tourniquet. With blood testing in sport these results have to be taken into consideration. PMID- 16037880 TI - Heart rate responses during a breath-holding competition in well-trained divers. AB - The diving response elicited by breath-holding (BH) and immersion mainly consists of bradycardia, decreased cardiac output, and peripheral vasoconstriction. These responses reduce oxygen consumption and thereby prolong the duration of the dive. They may also lead to cardiac arrhythmias or hypoxia, however, which in turn may play a role in the occurrence of syncope during BH. The aim of the present study was to analyze the cardiac responses to prolonged breath-holding in elite divers during a competition. Heart rate behaviour and the incidence of arrhythmia were recorded in 16 well-trained breath-hold divers (BHD) using a cardio-frequency meter (for 15 divers) and a Holter (for one diver) during maximal static breath holding. Anthropometric, spirometric, and training characteristics such as percentage of body fat, pulmonary volumes and years of BH training were also determined. Forced vital capacity (FVC) and forced expiratory volume in one second (FEV (1)) were higher than the predicted values (+7.7%, p<0.05 and+6.6%, p<0.05, respectively). During the static BH, divers presented apneic bradycardia (-44%) correlated with static BH times (p<0.05); this was associated with cardiac arrhythmias (supraventricular extrasystoles and ventricular extrasystoles) in the Holter-equipped subject. These results are in agreement with those obtained in laboratory conditions and confirm the existence of cardiac arrhythmias in well trained BHD. PMID- 16037881 TI - Evidence of major genes for plasma HDL, LDL cholesterol and triglyceride levels at baseline and in response to 20 weeks of endurance training: the HERITAGE Family Study. AB - This study assessed major gene effects for baseline HDL-C, LDL-C, TG, and their training responses (post-training minus baseline) in 527 individuals from 99 White families and 326 individuals from 113 Black families in the HERITAGE Family Study. The baseline phenotypes were adjusted for the effects of age and BMI, and the training response phenotypes were adjusted for the effects of age, BMI, and their respective baseline values, within each of the sex-by-generation-by-race groups, prior to genetic analyses. In Whites, we found that LDL-C at baseline and HDL-C training response were under influence of major recessive genes (accounting for 2--30 % of the variance) and multifactorial (polygenic and familial environmental) effects. Interactions of these major genes with sex, age, and BMI were tested, and found to be nonsignificant. In Blacks, we found that baseline HDL-C was influenced by a major dominant gene without a multifactorial component. This major gene effect accounted for 45 % of the variance, and exhibited no significant genotype-specific interactions with age, sex, and BMI. Evidence of major genes for the remaining phenotypes at baseline and in response to endurance training were not found in both races, though some were influenced by major effects that did not follow Mendelian expectations or were with ambiguous transmission from parents to offspring. In summary, major gene effects that influence baseline plasma HDL-C and LDL-C levels as well as changes in HDL-C levels in response to regular exercise were detected in the current study. PMID- 16037882 TI - Acute prolonged exercise reduces moderately oxidized LDL in healthy men. AB - We studied the effects of a 2-day walk exercise (6 h+6 h) on the serum concentration of circulating moderately oxidized LDL (LDL baseline conjugated dienes), lipids (total cholesterol, LDL cholesterol, HDL cholesterol, and triglyceride), antioxidants (alpha-tocopherol, gamma-tocopherol, beta-carotene, and ubiquinol-10), and antioxidant potential in serum (S-TRAP) and LDL (LDL-TRAP) in healthy well-trained men. The exercise was performed twice with an interval of 14 days. While 6 h walking the subjects drank 6 cl . kg (-1) water which contained either carbohydrate (CHO trial) or placebo (PLA trial). During the 2 day exercise the level of oxidized LDL decreased by 25 % (p=0.001) in the PLA trial. At the same time serum gamma-tocopherol decreased by 20 % (p=0.049), while the other measured antioxidants remained unchanged and the serum antioxidant potential increased by 22 % (p=0.018). Serum total cholesterol decreased by 3 % (p=0.017), serum triglycerides by 22 % (p=0.001), and LDL-cholesterol by 14 % (p=0.045). HDL cholesterol increased by 9 % (p=0.001). The results in the carbohydrate trial were similar to the ones in the PLA trial. The findings suggest that exercise of long duration but of low, non-exhaustive intensity decreases the concentration of circulating oxidized LDL simultaneously with an increase in serum antioxidant potential in healthy trained men. Carbohydrate ingestion during the exercise does not have any further effect on these changes. PMID- 16037883 TI - Cardiovascular responses to apnea during dynamic exercise. AB - Breath holding maneuvers induce hypoxia, hypercapnia, and various cardiovascular responses typically including increases in total peripheral resistance, mean arterial pressure (MAP) and decreases in heart rate (HR). During dynamic exercise these responses may have a generally negative impact on performance. Moreover, they deserve particular attention in cardiovascular risk subjects. In 26 healthy sport students we studied the HR and MAP effects induced by the combination of dynamic exercise (cycle ergometry, 30 W and 250 W) with 20 s of either respiratory arrest (mouth piece pressure held constant at 20 mm Hg), free breathing, or rebreathing, i. e. periods of unimpeded breathing leading to similar levels of hypercapnia and hypoxia as the respiratory arrest. The measurements yielded no major differences between the conditions of rebreathing and free breathing. In contrast, 20 s of apnea led to a marked increase in MAP and a HR depression at both levels of exercise intensity. Additionally, there was a delayed MAP recovery after this stimulus. The present findings show that breath holding has marked effects on MAP and HR during dynamic exercise, which are essentially independent of the resulting hypoxia and of increases in intrathoracic pressure. The key factor seems to be an increase in total peripheral resistance, probably including a vasoconstriction in the exercising muscles. PMID- 16037884 TI - Evidence of hepatic glucagon resistance associated with hepatic steatosis: reversal effect of training. AB - The present study was undertaken to test the hypothesis that a high-fat diet induced hepatic steatosis is associated with a reduction in hepatic glucose output (HGO) in response to a hyperglucagonemic infusion, and that this postulated state of hepatic glucagon resistance in high-fat fed rats is attenuated by concurrent exercise training. In four groups of anesthetized rats, glucagon (2 ug/kg/min iv) was infused over a period of 60 min to measure HGO. Two groups of rats were either fed a standard (SD) or a high-fat (HF; 42 % kcal) diet for eight weeks and were assigned either to a Sedentary (Sed) or a treadmill trained (TR) group. Training was initiated two weeks after the beginning of the diet protocol and was progressively increased over a period of 6 weeks reaching 60 min at 26 m/min, 10 % grade, for the last 3 weeks. The HF compared to the SD diet resulted in approximately 28 % higher (p < 0.01) liver triglyceride levels in Sed rats. This increase was completely prevented by the exercise training program in the HF-TR group. Plasma glucagon ( approximately 90,000 pg/ml) and insulin ( approximately 500 pmol/l) levels were increased to a similar extent in all four groups, with the exception of higher (p<0.05) insulin levels in SD-Sed group. Glucagon induced-hyperglycemia ( approximately 300 mg/dl) was higher (p<0.05) in the SD-Sed than in HF-Sed and SD-TR groups. Glucagon infusion resulted in a significantly (p<0.05) lower increase ( approximately 35 %) in HGO in HF-Sed compared to SD-Sed group. The lower level of HGO in HF-Sed compared to SD-Sed rats was observed whether HGO was measured after 25, 40, or 60 min of glucagon infusion. Exercise training in HF fed rats resulted in a significant (p<0.05) attenuation (50 %) of the state of HF-induced glucagon resistance. Comparisons of all individual liver triglyceride and 60-min HGO values revealed that liver triglyceride values were highly (p<0.001) predictive of the decreased glucagon action on HGO (R= -0.849). The present results indicate that the feeding of a high-fat diet induces a state of hepatic glucagon resistance, which is partially attenuated by concurrent exercise training. It is suggested that liver lipid infiltration may interfere with the action of glucagon, thus inducing glucagon resistance in liver. PMID- 16037885 TI - Is there an association between ACE and CKMM polymorphisms and cycling performance status during 3-week races? AB - In this paper, we examine the association between polymorphisms of the angiotensin-converting enzyme (ACE) and muscle-specific creatine kinase (CKMM) genes, and the actual performance status observed in professional cyclists capable of completing a classic tour stage race such as the Giro d'Italia, Tour de France, or Vuelta a Espana. To accomplish this, we compared the frequencies of the ACE and CKMM genotypes/alleles in 50 top-level Spanish professional cyclists that have completed at least one of these events to 119 sedentary controls, and 27 elite (Olympic-class) Spanish runners. The genetic polymorphism at the CK-MM locus was detected with the NcoI restriction endonuclease. The results of our study showed that the proportion of the DD genotype was higher in cyclists (50.0 %) than in the other two groups (p<0.05), the proportion of the ID genotype was higher in controls (46.2 %) than in the other two groups (p<0.05), and the proportion of the II genotype was higher in runners (40.7 %) than in the other two groups (p<0.05). The proportion of the D allele was higher in both cyclists (65.0 %) and controls (57.6 %) than in runners (46.3 %) (p<0.001), whereas the proportion of the I allele was higher in runners than in the other two groups (p<0.001). No statistical differences were found for CKK-MM- NcoI. We conclude that in top-level professional cyclists capable of completing a classic 3-wk tour race, the frequency distribution of the D allele and the DD genotype seems to be higher than in other endurance athletes such as elite runners (in whom the I allele is especially frequent). PMID- 16037886 TI - Endurance running ability at adolescence as a predictor of blood pressure levels and hypertension in men: a 25-year follow-up study. AB - The aim was to study whether aerobic fitness measured by a maximal endurance running test at adolescence predicts prevalence of hypertension or blood pressure levels in adulthood. From the 413 (197 slow runners and 216 fast runners) participating in a 2000-meter running test at adolescence in 1976 and responding to a health and fitness questionnaire in 2001, 29 subjects (15 very slow runners and 14 very fast runners) participated in a clinical follow-up study in 2001. Compared to those who were fast runners in adolescence, those who were slow runners tended to have higher age-adjusted risk of hypertension at follow-up (OR 2.7, 95 % CI 0.9 to 7.5; p=0.07). The result persisted after further adjustment for body mass index at follow-up (OR 2.9, 95 % CI 1.0 to 8.3; p=0.05). Diastolic blood pressure was higher for very slow runners at adolescence compared to very fast runners, the age-adjusted mean diastolic blood pressure being 90 mm Hg (95 % CI 86 to 93) vs. 83 mm Hg (95 % CI 80 to 87), age-adjusted p=0.013. High endurance type fitness in adolescence predicts low risk of hypertension and low resting diastolic blood pressure levels in adult men. PMID- 16037887 TI - Validity of a V.O2 max prediction equation of the 2-km walk test in female seniors. AB - Walking is a useful exercise mode for most adults due to its general ease, acceptability, and safety. Therefore, many field tests based on performance in walking have been developed to predict V.O (2 max). Even if these tests are much easier to perform than laboratory tests, field tests have to be valid. The objective of the paper was to explore the accuracy and bias of a V.O (2 max) prediction equation of the 2-km Walk Test, in an active female senior group (n=18, mean age: 66.1+/-4.4). V.O (2 max) (l . min (-1)) was measured during cycle ergometry by direct gas analysis from a maximal test (step: 30 W, time: 2 min 30). V.O (2 max) related to body mass was then calculated (ml . min (-1) . kg (-1)). Subjects completed also the 2-km Walk Test (UKK Institute). V.O (2 max) (ml . min (-1) . kg (-1)) was then predicted from age, sex, body mass index, heart rate, and walking time measured during the 2-km Walk Test. Predicted V.O (2 max) and measured V.O (2 max) were highly correlated (r=0.63, p<0.01). Predicted V.O (2 max) (20.5+/-6.1 ml . min (-1) . kg (-1)) was not significantly different from measured V.O (2 max) (18.7+/-3.4 ml . min (-1) . kg (-1)). Prediction equation bias with its 95 % limits of agreement was - 1.8+/-4.8 ml . min (-1) . kg (-1) with a coefficient of variation of 24.2 %. In an active female senior population, the 2-km Walk Test offers a fairly accurate V.O (2 max) prediction. The training and learning effects can be neglected because when the test was repeated no significant bias was observed between the two trials. PMID- 16037888 TI - Bone mineral density and degenerative changes of the lumbar spine in former elite athletes. AB - The aim of this study was to assess bone mineral density (BMD) and degenerative changes in the lumbar spine in male former elite athletes participating in different track and field disciplines and to determine the influence of body composition and degenerative changes on BMD. One hundred and fifty-nine former male elite athletes (40 throwers, 97 jumpers, 22 endurance athletes) were studied. Anthropometric (age, body mass index [BMI]) and sport-specific data (personal best, intensity, duration, and time since termination of competitive sports career as well as current sporting activity) were collected. Degenerative changes of the lumbar spine in lateral view were evaluated by using the Kellgren and Lawrence Score. Bone mineral density of the lumbar spine was measured in an anterior-posterior view with dual energy X-ray absorptiometry (DEXA, T-score). Throwers had a higher body mass index than jumpers and endurance athletes. Throwers and jumpers had higher BMD (T-LWS) than endurance athletes. Bivariate analysis revealed a negative correlation of BMD (T-score) with age and a positive correlation with BMD and Kellgren score (p<0.05). Even after multiple adjustment for confounders lumbar spine BMD is significantly higher in throwers, pole vaulters, and long- and triple jumpers than in marathon athletes. Different types of mechanical loading caused by sporting activities seem to influence the BMD of the lumbar spine, even if different body constitutions (i.e. BMI) and age, training history, and degenerative changes in the lumbar spine of former throwers, jumpers, and endurance athletes are taken into consideration. PMID- 16037889 TI - The effects of an ankle destabilization device on muscular activity while walking. AB - Chronic Ankle Instability sprain causes are unclear and many factors or mechanisms may contribute to recurrence of this injury. The aim of the study was to investigate how an ankle destabilization device affects the EMG patterns of the ankle muscles during ankle stabilization against inversion. The left foot was equipped with a mechanical device mounted under the heel of the shoe. This mechanical device induces subtalar joint destabilization necessitating the control of ankle muscles. Surface electrodes were placed over the tibialis anterior, the peroneus longus, the peroneus brevis, the gastrocnemius lateral, and the gastrocnemius medial. Nine healthy subjects (mean age 37+/-12 yr; mean mass 68+/-17 kg; mean height 1.73+/-0.7 m) were instructed to walk normally along a tape fixed on the floor. The ankle destabilization device altered the walking pattern of all subjects. More specifically, the walking pattern is disturbed resulting in higher amplitude of the EMG activity of the peroneal muscles and the Tibialis Anterior and anticipatory reactions in the peroneal muscles. The results suggest that the ankle destabilization device could be beneficial for rehabilitation programs especially during the training of walking. Using this material may help to a specific reinforcement of muscles involved in anti inversion ankle movement. PMID- 16037890 TI - Intracyclic velocity variations and arm coordination during exhaustive exercise in front crawl stroke. AB - The purpose of this study was to analyse the effects of an exhaustive exercise on arm coordination and intracyclic velocity variations (IVV) to better understand the ways in which they are modified under fatigue conditions. Seventeen competitive swimmers performed a 200-m all-out test and a set of two 25-m (before and after the 200-m) at maximal intensity to measure stroking parameters, IVV, and the relative duration of the different parts of the stroke cycle and identify the model of arm coordination by using the index of coordination (IdC). Results showed an increase in the relative duration of the propulsive time, which induced a change in arm coordination as fatigue developed in relation to a decrease in stroke length (SL) and stroke rate (SR) (p<0.05). The evolution of IdC corresponds to a reduction of the non-propulsive lag time between the two arms' propulsive actions. Despite these modifications, IVV were not significantly modified (p<0.05). The present results highlighted that IdC and relative durations of each part of the cycle (particularly the pull phase) could be assessed to complete the "SL x SR" model and to partly understand the technique modifications under fatigue condition. PMID- 16037891 TI - Delayed latency of peroneal reflex to sudden inversion with ankle taping or bracing. AB - The purpose of the present study was to examine the effects of ankle taping and bracing based on the peroneal reflex in the hypermobile and normal ankle joints with and without history of ankle injury. Thirty-six ankle joints of 18 collegiate American football athletes with and without previous history of injury were studied. The angle of talar tilt (TT) was measured by stress radiograph for classifying normal (TT5 degrees ) ankles. They were tested with taping, bracing, and without any supports as a control. The latency of peroneus longus muscle was measured by a sudden inversion of 25 degrees using surface EMG signals. The results of the present study show no significant three-way Group (hypermobile or normal ankles) by History (previously injured or uninjured ankles) by Condition (control, taping, or bracing) interaction, while Condition main effect was significant (p<0.05). There were significant differences between control (80.8 ms) and taping (83.8 ms, p<0.01), between control and bracing (83.0 ms, p<0.05), but not between taping and bracing (p>0.05). In conclusion, ankle taping and bracing delayed the peroneal reflex latency not only for hypermobile ankles and/or injured ankle joints but also for intact ankle joints. PMID- 16037892 TI - The effects of dehydration on brain volume -- preliminary results. AB - In adults the cranium is a rigid bony vault of fixed size and therefore the intra cranial volume is a constant which equals the sum of the volume of the brain, the intra-cranial volume of CSF and the intra-cranial volume of blood. There can be marked changes in the volumes of these three intra-cranial compartments which may influence susceptibility to brain damage after head injury. This is the first study to investigate the relationship between dehydration and changes in the volume of the brain and the cerebral ventricles. Six healthy control subjects underwent magnetic resonance imaging of the brain before and after a period of exercise in an environmental chamber. The subjects lost between 2.1 % and 2.6 % of their body mass due to water loss through sweating. We found a correlation between the degree of dehydration and the change in ventricular volume (r=0.932, p=0.007). The changes in ventricular volume caused by dehydration were much larger than those seen in day-to-day fluctuations in a normally hydrated healthy control subject. PMID- 16037893 TI - Disordered eating in Japanese and Chinese female runners, rhythmic gymnasts and gymnasts. AB - Eating Attitude Test-26 (EAT-26) and self-administered questionnaires were used to survey eating attitudes and menstrual irregularity of Japanese (J) and Chinese (C) collegiate female runners (RUNs), rhythmic gymnasts (RGYMs), gymnasts (GYMs), and nonathletic controls (NAs). Athletes were recruited from several outstanding colleges for physical education and sport in each country. The prevalence of disordered eating (DE) was significantly higher in Japanese athletes (21% in JRUNs, 19% in JRGYMs, and 15% in JGYMs) than Chinese athletes (4% in CRUNs, 2% in CRGYMs, and none in CGYMs). Also, the prevalence of amenorrhea was very low in CRUNs (1%), CRGYMs (0%), and CGYMs (0%), as compared with their respective Japanese counterparts (JRUNs 22%, JRGYMs 10%, JGYMs 8%). There was no significant difference in the prevalence of DE and amenorrhea between Japanese and Chinese NAs. Multivariate logistic regression analysis using 10 independent variables regarding anthropometrics, athletics, weight and diet concern, and nationality indicated that high frequency of dieting during a lifetime, high athletic level and being Japanese were significant risks for the development of DE in athletes. It is concluded that the low prevalence of DE in the Chinese in this study is partly explained by the lack of socioculturally- and socioeconomically-imposed desire to be thin as well as by the low frequency of dieting during their lifetime. PMID- 16037894 TI - Modelling the relationships between training, anxiety, and fatigue in elite athletes. AB - This study investigated the effects of 40-week training on anxiety and perceived fatigue in four elite triathletes. Anxiety and perceived fatigue were self reported by the subjects twice a week by the way of a specific questionnaire and were linked by a mathematical model to the training loads calculated from the exercise heart rate. A significant relationship (r=0.32; p<0.001) between the training loads and anxiety was identified using a two-component model: a first, negative (i.e., anxiety decreased) short-term (tau (1)=23 days) function and a second, positive long-term (tau (2)=59 days) function. The relationship between the training loads and perceived fatigue was significant (r=0.30; p<0.001), with one negative function (tau (1)=4 days). This mathematical model can potentially describe the relationships between training loads and anxiety or perceived fatigue and may improve both the adjustment of the duration of tapering and the early detection of staleness. PMID- 16037895 TI - Energy turnover at the Race Across AMerica (RAAM) - a case report. AB - We report about energy intake and energy expenditure in an official finisher of the Race Across AMerica (RAAM) in 2003. Energy intake from nutrition was continuously recorded. Energy expenditure was measured by continuous heart rate recording with a portable heart rate monitor POLAR S710 to estimate energy expenditure during physical activity. Our athlete (33 years, 179 cm, 73 kg, VO (2)max 60 ml . min (-1) . kg (-1), lactate threshold at 77%VO (2)max) finished the 4701-km cycling race with altitude differences amounting to 25,826 meters in 9 days 16 hours and 45 minutes in 4th place. He completed 470+/-72.9 km (372-541 km) per day with 2,582+/-1,576 (683-5,047) meters of altitude. During the whole race, he expended a total energy of 179,650 kcal with 17,965+/-2165 (15,100 - 23,280) kcal per day. Total energy intake was 96,124 kcal with an average of 9,612+/-1,500 (7,513-12,735) kcal per day. Of total ingested calories, 75.2% derived from carbohydrates, 16.2% from fat, and 8.6% from protein. He ingested an energy of 9,612+/-1,550 (7,513 - 12,735) kcal daily, consisting of 1,814+/-310 (1,336 - 2,354) g of carbohydrates, 172+/-47 (88-251) g of fat, and 207+/-52 (128 286) g of protein. The average daily energy deficiency amounted to 8,352+/-2,523 (4,425-13,631) kcal. A total deficiency of 83,526 kcal resulted after the race while the athlete lost 5 kg of body weight. These results provide data about energy intake and energy expenditure in the RAAM for future athletes, nutritionists and coaches. Further investigation should be performed in order to determine whether either muscle mass or body fat will be lost in extreme endurance cycling. PMID- 16037896 TI - Letter to the editors - Re: Heinicke K, Heinicke I, Schmidt W, Wolfarth B. A three-week traditional altitude training increases hemoglobin mass and red cell volume in elite biathlon athletes. Int J Sports Med 2005; 26: 350-355. PMID- 16037898 TI - Haematological testing and antidoping policies - Re: Robinson N, Schattenberg L, Zorzoli M, Mangin P, Saugy M. Haematological analysis conducted at the departure of the Tour de France 2001. Int J Sports Med 2005; 26: 200-207. PMID- 16037900 TI - [Systemic therapy of operable carcinoma of the breast]. PMID- 16037901 TI - [Surveillance in women with early breast cancer, systematic versus symptom guided follow-up]. AB - Nearly all national (AGO, DKG) and international guide lines (e. g. ASCO) for follow-up of breast cancer patients do not explicitly recommend regular laboratory and radiological/ultrasound screening procedures. According to these guide lines, follow-up should be focused on the breast, only patients with possibly tumour related symptoms should be screened for metastatic disease. The rejection of more time-consuming and costlier follow-up examinations remains a contradiction to established follow-up guide lines for other solid tumours. In addition, treatment options for metastatic breast cancer disease have improved continuously over the last years. However, treatment options are considerably limited in advanced disease, if e. g. symptoms like dyspnoea or jaundice are already present at first diagnosis of metastatic disease. Therefore we will review available data of older studies as well as discuss arguments for a systematic surveillance in high-risk breast cancer patients. Overall, symptom guided follow-up seems to be adequate for patients with small primary tumours, no lymph node involvement and therefore a high curative probability, whereas in the authors' opinion systematic surveillance should be recommended for high risk patients even in the absence of symptoms. All patients, however, should be fully informed about the possibility of metastatic disease development and should be enabled to select the quality of their postoperative follow-up. PMID- 16037902 TI - [Mammographic breast density and breast cancer risk during HRT]. AB - Higher breast density leads to a higher risk of breast-cancer coming along with a reduced sensitivity of mammography, the most important method for early diagnosis of breast cancer. HRT leads to an increase in breast density in up to (1/3) of treated women. Combined regimes of estrogen-progestin show a stronger influence than estrogen only. Changes of breast density appear almost entirely within the first year of administration and seem to be reversible after suspension of treatment within a few weeks. A possible solution of the dilemma is to perform mammography in asymptomatic premenopausal women during the first half of the menstrual cycle. In women with increased breast density using HRT it is recommended to modify the combination of hormones, change the application mode, or suspend HRT for a short period of 3 weeks prior to mammography. In symptomatic women with dense breasts, additive breast ultrasound is strongly recommended to reduce the otherwise high rate of missed breast cancers. PMID- 16037903 TI - [Metastatic breast cancer: options for metastasectomy]. AB - Usually, the therapy of metastatic breast cancer consists of chemotherapy or endocrine therapy, because even in the case of isolated metastases in one organ, diffuse tumor cell dissemination exists, so that local surgical treatment does not seem sensible. Particurlarly in patients with hepatic or pulmonary metastases the indication for hepatic or pulmonary metastasectomy should be individualized, as hepatic or pulmonary metastases usually develop during a phase of disease, when extrahepatic or -pulmonary metastases also can be detected. Only in patients with long disease-free interval, with isolated hepatic or pulmonary metastases, and the possibility of R0-resection is hepatic or pulmonary metastasectomy a therapeutic option in selected cases. PMID- 16037904 TI - [Estrogen receptor Beta isoforms -- functions and clinical relevance in breast cancer]. AB - Estrogen receptors (ER) are hormone regulated transcription factors which mediate the cellular estrogen response. So far two types of ER could be identified, ERalpha and ERbeta. ERalpha plays an important role as a predictive factor for the effectiveness of endocrine agents in the treatment of breast cancer. Patients with ERalpha positive tumors receive adjuvant endocrine therapy. Recent findings indicate a crucial role of ERbeta and its isoforms for the prognosis of breast cancer. This paper reviews the current knowledge on the functions of ERbeta and its splice variants. PMID- 16037905 TI - [Therapy-induced leukemia -- an underestimated complication of antineoplastic chemotherapy?]. AB - Alkylating agents and topoisomerase II inhibitors are mutagenic cytotoxic drugs which may induce therapy-related acute myeloid leukemia or myelodysplasia. The frequency of these complications depends on the type of agent used, its dosage, and the duration of treatment. Commonly used protocols for adjuvant chemotherapy of breast cancer, such as the CMF or AC protocols, are associated with a leukemia rate of 0.2 to 0.5 % after 10 years. Intensification of chemotherapy or additional radiotherapy lead to a significant increase in the incidence of leukemia. The prognosis of therapy-related leukemia is dismal. Therefore, it appears mandatory to restrict adjuvant chemotherapy to those patients who are the most likely to benefit from it. PMID- 16037906 TI - [Management of HIV-positive pregnancies -- results from a retrospective study]. AB - OBJECTIVE: The management of HIV-positive pregnancies was investigated in conjunction to pre-, peri and postpartal complications and the HIV transmission rate. PATIENTS AND METHODS: Retrospective study of 88 HIV-positive patients who were delivered at the Dept. of Obstetrics and Gynaecology during 1.1.1997 31.12.2001. RESULTS: HIV-positive patients showed significantly more prepartal complications, compared to control group. Low CD4-cell count (< or = 200/microl) or high viral load (> 10 000 HIV-copies/ml) was not associated with increased risk for transmission relevant complications. The overall HIV-transmission rate was 3.4 % (3/89 newborns; with ART 2.5 % [2/85], without ART 33.3 % [1/3]). The transmission rate increased with complications during pregnancy (7.7 % [2/26] vs. 1.6 % [1/61]). Newborns delivered < or = 35 (th) week of gestation showed a transmission rate of 5.3 % compared to 2.9 % of newborns delivered after the 35 (th) week of gestation. 98 % of the patients were delivered by cesarean section (primary: n = 71, prior: n = 15), spontaneously: n = 2). 97 % of patients (85/88) were treated with antiretroviral therapy (ART). No differences were found in the postpartal complication rate of HIV-positive to -negative patients. None of the newborns was breast fed. CONCLUSIONS: Treatment of this risk-pregnancies in HIV experienced centers significantly reduces the risk of HIV transmission. PMID- 16037911 TI - Seasonal recruitment in case-control studies by Stallard and Tombs, Statistics in Medicine 2004; 23:3193-3207. PMID- 16037907 TI - [Mediastinal large B-cell lymphoma with symptomatic superior vena cava syndrome in a patient with bichorial twin pregnancy in the 26th week of gestation: peri and postpartal management -- a case report]. AB - OBJECTIVE: Non-Hodgkin lymphoma (NHL) is rarely observed during pregnancy. The clinical behavior of this malignancy does not differ significantly from that outside of the setting of pregnancy. Antineoplastic chemotherapy is usually given during the second and third trimester. However, irradiation is another therapeutic option. The teratogenic potential limits its use in pregnancy. Finding an appropriate therapeutic management in an emergency setting is therefore difficult. CASE REPORT: In this report, we describe the case of a 31 year-old gravida three, para two, in whom a mediastinal large B-cell lymphoma with symptomatic superior vena cava syndrome was diagnosed in a bichorial twin pregnancy in the 26 (th) week of gestation. After premature delivery by caesarean section at 26 + 0 weeks gestation the patient was immediately submitted to mediastinal irradiation. The clinical symptoms resolved and adjuvant CHOEP chemotherapy was instituted. Chemotherapy was well tolerated and a partial remission was observed after 4 cycles. The neonatological follow-up was uneventful. DISCUSSION: To our knowledge, this is the first case in the literature of a patient with bichorial twin pregnancy with large cell mediastinal NHL and symptomatic superior vena cava syndrome who underwent irradiation after caesarean section because of life-threatening medical condition. PMID- 16037912 TI - Sonographic assessment of urinary retention in multiple system atrophy and idiopathic Parkinson's disease. AB - Sonography of the bladder was performed before and after voluntary voiding in 20 subjects with possible/probable multiple system atrophy (MSA), 20 patients with idiopathic Parkinson's disease (PD), and 20 healthy controls. Mean residual volume was 173 ml (SD, 194) in MSA and significantly increased compared to both PD (21 ml; SD, 49) and controls (13 ml; SD, 12). Incomplete voiding (>100 ml of residual volume) was found in 11 subjects with MSA but only in 1 patient with PD. Positive predictive value of increased residual volume for MSA was 91.6% in this study, whereas negative predictive value was only 67.8. Bladder sonography is an objective, simple, and safe tool that allows one to screen for urinary retention, which is highly suggestive but incompletely sensitive for MSA. Because sonography is easily accessible and rapidly performed, it is feasible for routine assessment of atypical Parkinsonism. PMID- 16037913 TI - Efficient internal pallidal stimulation in Gilles de la Tourette syndrome: a case report. AB - The usefulness of deep brain stimulation (DBS) of thalamic nuclei in Gilles de la Tourette syndrome (GTS) has recently been advocated. We report on a 14-month follow-up study of a patient with intractable GTS in whom bilateral DBS of the internal globus pallidus was carried out. Tic frequency per minute decreased by 73% in the postoperative phase and in particular the vocal tics became less intense. Pronation/ supination bradykinesia of the left extremities was a well tolerated, permanent side effect. Pallidal DBS could become a valuable rescue therapy for otherwise intractable GTS. PMID- 16037914 TI - Paraneoplastic chorea with leukoencephalopathy presenting with obsessive compulsive and behavioral disorder. AB - Chorea is a rare manifestation of paraneoplastic disease and is associated with CV2/CRMP-5 antibodies. Obsessive-compulsive disorder and large-scale white matter abnormalities on MRI have not been previously reported in association with these antibodies. We report on a case of CV2 paraneoplastic syndrome with obsessive compulsive behavior preceding the motor manifestations of chorea with associated leukoencephalopathy on MRI. The literature on paraneoplastic chorea is reviewed. PMID- 16037915 TI - Clinical and neurophysiologic spectrum of orthostatic tremor: case series of 26 subjects. AB - Orthostatic tremor (OT) is a condition described as high-frequency tremors predominantly in the legs and trunk, which are present not only in the standing position but also during isometric contraction of the limb muscles. This report is one of the largest OT series describing clinical and neurophysiologic findings in 26 subjects with OT. The main findings included 13.0 to 18.6 Hz leg tremors while standing with varied patterns of phase relationships between the antagonists of the ipsilateral leg and between the homologous muscles of the contralateral leg, short latency tremor onset upon standing with abrupt cessation after sitting, coexistence of tremors in the cranial structures and the arms, and sense of unsteadiness without actual falls. Although the oscillator of OT is most likely located in the brainstem, cerebral cortex, basal ganglia, and cerebellum may also be involved in its pathogenesis. PMID- 16037916 TI - Sway patterns in orthostatic tremor: impairment of postural control mechanisms. AB - Sway parameters in orthostatic tremor (OT) patients were compared with age matched controls. The effects of vision (eyes open or closed), stance width (feet apart or together), and external support (with or without) on sway and 14-18 Hz energy were measured. Sway in OT patients decreased in the presence of each of the stabilizing factors but the extent of benefit obtained by OT patients was significantly less than controls for support for the sagittal plane (22% vs. 42% decrease; P < 0.01) and feet apart in the mediolateral plane (38% vs. 65% decrease; P < 0.01). Three patients with proprioceptive loss also had a reduced response to these factors. Energy in the 14-18 Hz range did not always change in parallel with sway levels. Vision suppresses activity at the OT frequency in patients. Postural control mechanisms in OT remain responsive to postural conditions, but patients differ significantly from normals in the degree of their responsiveness. OT appears to have disruption of the normal generation or processing of proprioceptive signals as one of its important components. PMID- 16037917 TI - Changes induced by levodopa and subthalamic nucleus stimulation on parkinsonian speech. AB - Levodopa (L-dopa) and subthalamic nucleus (STN) stimulation treatments have been associated with both improvement and exacerbation of dysarthria in Parkinson's disease (PD). We report four cases illustrating variant responses of dysarthria to dopaminergic and STN stimulation therapies. Patients' motor disability and dysarthria were perceptually rated by the Unified Parkinson's Disease Rating Scale (UPDRS) in four conditions according to medication and STN stimulation. Dedicated software packages allowed acquisition and analysis of acoustic recordings. Case 1, who had a severe off period aphonia, experienced improvement of speech induced by both levodopa and STN stimulation. In Case 2, both treatments worsened speech due to the appearance of dyskinesias. Case 3 had a dysarthria exacerbation induced by STN stimulation with parameters above optimal levels, interpreted as current diffusion from the STN to corticobulbar fibers. In Case 4, dysarthria exacerbation occurred with stimulation at an electrode contact located caudally to the target, also arguing for current diffusion as a potential mechanism of speech worsening. The presented cases demonstrated variant effects in relation to L-dopa and STN stimulation on speech. It seems that motor speech subcomponents can be improved like other limb motor aspect, but that complex coordination of all speech anatomical substrates is not responsive to STN stimulation. These hypotheses may be helpful for better understanding and management of STN stimulation effects on motor speech and skeleton-motor subsystems. PMID- 16037918 TI - Parkinson's disease, stroke, and related epidemiology. AB - We investigated the prevalence of cerebrovascular disease and other comorbidities in Parkinson's disease (PD) patients compared to the general population. Five hundred PD patients were chosen randomly from one author's (A.H.R.) database. Age and sex-matched controls were derived from 270 patients with essential tremor from the same database and from 490 patients in a general practitioner's database. Age, hypertensive status, smoking status, coronary artery disease, orthostatic hypotension, diabetes mellitus, and symptomatic cerebrovascular disease (stroke or transient ischemic attack) were assessed. Statistical analysis was performed using Pearson chi(2) testing and binary logistic regression analysis. The prevalence of coronary artery disease, hypertension, diabetes mellitus, and orthostatic hypotension was similar among groups. The PD group had more patients who never smoked and less current smokers than the other groups. While there were similar frequencies of symptomatic cerebrovascular disease among groups, the prevalence of stroke was lower in PD patients. This difference disappeared upon stratification into groups based on smoking status and in the addition of smoking as a covariate in the multivariate analysis. Diminished smoking in PD patients likely plays a role in our finding of decreased stroke in patients with PD. Increased access to appropriate neurological care and subsequent prevention of stroke after a warning transient ischemic attack may also play a role, as may diminished levels of excitotoxic neurotransmitters in PD patients. PMID- 16037919 TI - Re: Patient-based outcomes of cervical dystonia: a review of rating scales. PMID- 16037920 TI - Hedonistic homeostatic dysregulation in Parkinson's disease and excess dopamine replacement therapy. PMID- 16037921 TI - Facial palsy-induced blepharospasm relieved by a Bangerter foil. PMID- 16037922 TI - Burden of disease related to Parkinson's disease in Spain in the year 2000. AB - We measured the burden caused by Parkinson's disease (PD) in Spain during the year 2000 and compared it against PD burden worldwide and in the European A subregion. Burden of disease (BoD) is an important factor in health policy. Disability-adjusted life years (DALY) as a measure of BoD is the result of adding years of life lost (YLL) and years lived with disability (YLD). The burden of PD (BPD) has not been studied in Spain. YLL were obtained from the Spanish death certificates and YLD from the estimated number of incident PD cases and the average PD duration. PD disability was calculated, using the Disability Weights for Diseases in the Netherlands. Prior PD DALY data for Europe and the world were obtained from the 2001 World Health Organization World Health Report. A discount rate of 3% and age-weighting modulation factor with K = 1 were used. In Spain, PD generated 67,582 DALY, comprising 6,351 (9.4%) YLL and 61,231 (90.6%) YLD. Most PD DALY (57.5%) occurred in the population 60 to 74 years of age. When PD DALY estimates were adjusted using the world population in 2000, Spain registered a PD DALY rate of 84 per 100,000 population, higher than both the world and European A subregion rates (24 and 35 per 100,000 population, respectively). PD burden in Spain in 2000 was high, with disability being the major contributing factor. Although BPD in Spain was greater than both world and European A subregion BPD, these differences should nevertheless be interpreted with caution. PMID- 16037924 TI - Suboptimal medication adherence in Parkinson's disease. AB - Patients take less medication than prescribed in many disease areas but evidence for suboptimal therapy adherence in Parkinson's disease (PD) is limited. A single center observational study of antiparkinsonian medication was undertaken using electronic monitoring (MEMS; Aardex, Zug, Switzerland) over 3 months. Of 68 patients approached, 6 declined and 8 dropped out, leaving 54 patients (taking 117 preparations) with available data. Poorer compliance was associated significantly with younger age, with taking more antiparkinsonian tablets per day, with higher depression scores, and with poorer quality of life. Of the 54 evaluable patients, 11 (20%) had average total compliance of under 80% (underusers) and 43 (80%) had average total compliance of over 80% (satisfactory adherence). Underusers had median total compliance of 65% (interquartile range, 37-74) versus 98% (interquartile range, 93-102) in the satisfactory adherence group. Timing compliance (number of doses taken in the correct time interval) was poor in both underusers (median, 11%; interquartile range, 2-20) and those with satisfactory adherence (median, 25%; interquartile range, 11-73). In conclusion, poorer compliance is associated with younger age, depression, and more tablets per day, and one-fifth of PD patients underuse medication. Consideration of drug therapy adherence has implications in the management of PD. PMID- 16037925 TI - Comment on Sagel et al. PMID- 16037926 TI - The factors that influence the elution order for the resolution of amino acids on vancomycin phase using the polar-organic mobile phases after their pre-column derivatization with electrophilic reagents. AB - A variety of amino acids were enantioresolved on a vancomycin bonded chiral phase using the polar-organic mobile phases after their pre-column derivatization with electrophilic reagents in alkaline medium. The resolution was highly dependent on the analyte's structure and was enhanced as the aromatic side-chain group on the skeleton of analyte for pi-pi interaction with the chiral selector became available. The steric hindrance resulting from the bulky side-chain group on the analyte also affected the resolution. Elution reversal, not found on the teicoplanin phase under the same chromatographic conditions, was possible through altering the type of reagent used in the derivatizing reaction (e.g. 2,4 difluorophenylisothiocyanate to 2,4-difluorophenylisocyanate). It is believed that the steric hindrance, as a result of the bulky sulfur atom in reagents such as methylisothiocyanate and others examined in this study, was responsible for the reversed elution order. The bulkiness of the substituent on the aromatic ring of derivatizing reagents (i.e. 2,3- and 3,5-dichlorophenylisothiocyanate) was observed to affect the resolution and alter the elution order as well. PMID- 16037927 TI - Synthesis and evaluation of a new biselector s-triazine based chiral stationary phase for enantioselective HPLC: potentiality of the approach and perspectives. AB - A s-triazine scaffold bearing a free and a protected amino group was used for connecting two different chiral auxiliaries, whose enantiodiscriminating capabilities in enantioselective chromatography are well documented. A biselector system was synthesized and, after linkage to silica gel, used for the chromatographic resolution of different racemic analytes, chosen among the compounds resolved by the two different isolated chiral auxiliaries. The obtained chromatographic results were compared with literature data related to the use of the two chiral stationary phases (CSPs) whose chiral moieties constitute the biselector CSP, allowing us to gain useful information about potentialities and limitations of this approach for obtaining independent CSPs having broader applicability with respect to the classical independent monoselector CSPs. PMID- 16037928 TI - Effect of alcohols and temperature on the direct chiral resolutions of fipronil, isocarbophos and carfentrazone-ethyl. AB - The enantiomeric separations of three pesticides fipronil (asymmetric nitrogen), isocarbophos (asymmetric phosphorus) and carfentrazone-ethyl (asymmetric carbon) were studied on cellulose-tri(3,5-dimethylphenylcarbamate) chiral stationary phase using high-performance liquid chromatography under normal phase. The mobile phase was n-hexane with alcohols including ethanol, n-propanol, iso-propanol, n butanol and iso-butanol as polar modifiers. The flow rate was 1.0 mL/min with UV detection at 280, 225 and 230 nm for fipronil, isocarbophos and carfentrazone ethyl respectively. The influence of the modifiers and their volume content and temperature from 0 to 50 degrees C on the separations was investigated. The chiral stationary phase showed excellent stereoselectivity for the two enantiomers of fipronil and isocarbophos and certain chiral recognition for carfentrazone-ethyl. Iso-propanol was more suitable for the chiral separation of isocarbophos and carfentrazone-ethyl, and iso-butanol was better for fipronil. The resolutions increased with the decreasing modifier content and temperature for all the three chiral pesticides. PMID- 16037930 TI - Enantiorecognition on solid chiral selectors using microbatch technology: an example of limitation in case of strong association in the racemate. AB - When they were independently tested, the enantiomers of N,N'-bis(salicylidene) trans-1,2-cyclohexanediamine showed a large difference in adsorption on new chiral selectors using microbatch technology. Surprisingly, when these enantiomers were applied on the same supports as a racemic mixture, no discrimination was observed even though suitable adsorption existed. When a mixture enriched in one enantiomer (scalemic mixture) was applied, the resulting supernatant contained the racemic form and the enantiomer in excess was adsorbed on the support together with a part of racemate. This behavior, which militates in favor of a strong heterochiral dimer formation in the racemate, was revealed using microbatch technology but remained hidden on classical column chromatography on chiral support. Molecular dynamics calculations corroborate this hypothesis, showing a favorite binding mode of the heterochiral dimer, which is stabilized by various inter- and intramolecular interactions. Our findings may be considered as a new limitation of microbatch technology, but they may have some inference in case of chiral amplification using the N,N'-bis(salicylidene) trans-1,2-cyclohexanediamine enantiomers as chiral ligands. PMID- 16037929 TI - Column selection and method development for the separation of nucleoside phosphotriester diastereoisomers, new potential anti-viral drugs. Application to cellular extract analysis. AB - Analytical HPLC methods using derivatized cellulose and amylose chiral stationary phases used in normal and reversed-phase modes were developed for the diastereoisomeric separation of mononucleotide prodrugs (pronucleotides) of 3' azido-2',3'-dideoxythymidine (AZT). The resolutions were performed with two silica-based celluloses using normal and reversed-phase methodologies: Tris-3,5 dimethylphenylcarbamate (Chiralcel OD-H and Chiracel OD-RH) and Tris methylbenzoate (Chiralcel OJ and OJ-R). Two amyloses phases, Tris-3,5 dimethylphenylcarbamate (Chiralpak AD) and Tris-(S)-1-phenylethylcarbamate (Chiralpak AS), were used in normal-phase mode. Additionally, we developed separation using two stationary phases with immobilized cyclodextrins in reversed phase and polar-organic modes. The mobile phase and the chiral stationary phase were varied to achieve the best resolution. Different types and concentration of aliphatic alcohols, acetonitrile or water in the mobile phase were also tested for the different separation modes. An optimal baseline separation (Rs > 1.5) was readily obtained with all silica-based celluloses and amyloses using a normal phase methodology. The different columns gave complementary results in term of resolution. Limits of detection and quantification were 0.12-0.20 and 0.40-0.67 microm, respectively. This analytical method was applied in a preliminary study for the pronucleotide 2 quantification in cellular extract. PMID- 16037931 TI - Application of dimethyl-beta-cyclodextrin as a chiral selector in capillary electrophoresis for enantiomer separation of ephedrine and related compounds in some drugs. AB - Dimethyl-beta-cyclodextrin (DM-beta-CD) modified capillary electrophoresis has been developed for chiral separation of ephedrine and related compounds, such as (+/-)-norephedrine, (+/-)-N-methylephedrine, (+/-)-ephedrine and (+) pseudoephedrine. The influence of some crucial parameters such as buffer concentration, pH value, DM-beta-CD concentration, applied voltage and separation temperature on the separation was investigated. Under the optimum conditions, i.e. 40 mM DM-beta-CD in 75 mM Tris (pH 2.5) as the running electrolyte, separation voltage +25 kV and temperature 25 degrees C, a satisfactory separation of the enantiomers was accomplished. The detection limits (S/N = 3) ranged from 65 to 161 ng/mL and the linear range was 0.15 to 101.0 microg/mL for pressure injection. The present method was successfully applied for the analysis of a series of drugs such as anti-tussive, the drug for rheum, the drug for rhinitis and a Chinese traditional herbal medicine, Ephedrae herba (Ma-Huang in Chinese). The recoveries of ephedrine and related compounds in real samples ranged from 97.6 to 103.5%. This method is useful in the simple and rapid analysis of ephedrine derivatives in marketed products. PMID- 16037932 TI - Gas chromatographic quantification of free D-amino acids in higher vertebrates. AB - D-amino acids were determined in brain, body fluids (urine, blood coagulate, serum, plasma) and faeces of animals belonging to nine out of 11 taxonomic orders of vertebrates (Artiodactyla, Aves, Carnivora, Lagomorpha, Marsupalia, Osteichthyes, Primates, Rodentia, Tubilidentata). Free amino acids were isolated by means of cation exchangers and converted into volatile N(O)-perfluoroacylamino acid propyl esters. Derivatives of amino acids were separated into D- and L enantiomers using Chirasil-L-Val capillary columns and detected by selected ion monitoring mass spectrometry. Quantification of amino acids was achieved by comparison of analytes with amino acid standards using L-norleucine as internal standard. Large relative amounts of D-serine were determined in brains of mammals but not of birds. In body fluids the D-enantiomers of most proteinogenic L-amino acids were detected, largest absolute and relative amounts were found in urine. Therein quantities of D-Ala and D-Ser exceeded 50% relative to the L-enantiomers in many instances. Feeding animals with diet fortified with DL-Met resulted in excretion of almost racemic Met in urine. D-Amino acids were also abundant in faeces of rodents. The data confirm that d-amino acids are common in body fluids and certain tissues of vertebrates. PMID- 16037933 TI - Thin-layer chromatography separation of enantiomers of verapamil using macrocyclic antibiotic as a chiral selector. AB - Silica gel thin-layer chromatography plates impregnated with macrocyclic antibiotic, vancomycin, as chiral selector were prepared and used for the resolution of (+/-)-verapamil. A mobile phase system of acetonitrile-methanol water (15:2.5:2.5, v/v) was worked out systematically. The effects of chiral selector, temperature and pH on resolution were also studied. The spots were detected with iodine vapors and the detection limit was found to be 0.074 microg of each enantiomers. PMID- 16037935 TI - Spinocerebellar ataxia type 17: extension of phenotype with putaminal rim hyperintensity on magnetic resonance imaging. AB - We report on a 50-year-old woman who presented with an 8-year history of involuntary movements, unsteadiness, and cognitive decline. Examination revealed multidomain cognitive deficits, jerky ocular pursuit movements, hypometric saccades, gaze impersistence, dysarthria, upper limb dystonia, and widespread chorea. TATA-binding protein gene test revealed trinucleotide expansion allele sizes of 47 and 39 repeats, confirming the diagnosis of spinocerebellar ataxia type 17 (SCA-17). Magnetic resonance imaging (MRI) showed marked cerebellar atrophy and putaminal rim hyperintensity. This is the first case of SCA-17 reported to show MRI signal change in the basal ganglia, and extends the phenotypic manifestation of SCA-17. PMID- 16037936 TI - Clinical feature profile of spinocerebellar ataxia type 1-8 predicts genetically defined subtypes. AB - An increasing number of genetically defined types of spinocerebellar ataxia (SCA) have been reported in the past decade. Phenotype--genotype correlation studies have suggested a broad overlap between SCA types. The aim of the present study was to identify patterns of clinical features that were likely to distinguish between SCA types and to test the specificity and sensitivity of these signs and symptoms using a Bayesian classifier. In total, 127 patients from 50 families with SCA types 1 to 8 were examined using a worksheet with a panel of 33 symptoms and signs. By computing the probabilities of each trait for each SCA type, we rated the predictive value of each feature for each form of ataxia and then combined the probabilities for the entire panel of traits to construct a Bayesian classifier. Results of this analysis were summarized in a simpler, more operator based algorithm. Patients with SCA5, SCA6, and SCA8 demonstrated a predominant cerebellar syndrome, whereas patients with SCA1, SCA2, SCA3, SCA4, and SCA7 frequently had clinical features indicating an extracerebellar involvement. The Bayesian classifier predicted the SCA type in 78% of patients with sensitivities between 60 and 100% and specificities between 94 and 98.2%. The highest sensitivity to correctly predict the true SCA type was found for SCA5, SCA7, and SCA8. Sensitivities and specificities found in the present study validate the use of algorithms to help to prioritize specific SCA gene testing, which will help to reduce costs for gene testing. PMID- 16037937 TI - Heterotopic hindlimb allotransplantation in rats: an alternative model for immunological research in composite-tissue allotransplantation. AB - A nonfunctional heterotopic hindlimb allotransplantation model is presented, and a study was performed between this model and standard orthotopic hindlimb transplantation to compare operation and ischemia time, overall morbidity, and mortality rates. In this model, the skin component of the hindlimb was stripped away to the ankle level, and the remaining part along with the vascularized epigastric skin was transplanted heterotopically to the inguinal space of the recipient. No osteotomy and intramedullary fixation were performed. Thirty heterotopic and orthotopic hindlimb transplantations were performed in two groups. Each group involved 15 transplantations from inbred, male Brown Norway (BN; RT1n) weighing 150-200 g to Lewis rats (LEW; RT1(1)) weighing 250-300 g. The animals were followed up for 2 weeks under immunosuppression with cyclosporine A (16 mg/kg/day). Mean operation times for heterotopic and orthotopic hindlimb transplantations were 60 and 105 min, with ischemia times of 35 and 85 min, respectively. No animal deaths or major complications were encountered in heterotopic hindlimb transplantation during the follow-up period. Seroma formation was observed in one animal as the only minor complication. The mortality rate for orthotopic hindlimb transplantation was 26.7%, and there were minor complications in 35%, including infection, ulceration, and loss of rigid fixation. This model circumvents the disadvantages of osteotomy and intramedullary fixation, which may increase the risk of blood loss, embolus, and infection in immunosuppressed animals. It also helps avoid tension or kinking on the anastomosis due to inadequate judgment of the osteotomy level, distortion associated with loss of rigid fixation, and weight mismatch between donor and recipient. From the immunological point of view, insult to bone marrow is avoided, and a relatively constant amount of bone marrow is introduced to be used in chimerism-based tolerance studies. We recommend this model for composite tissue allotransplantation studies when functional recovery is not of primary importance. PMID- 16037938 TI - The role of adsorbed fibrinogen in platelet adhesion to polyurethane surfaces: a comparison of surface hydrophobicity, protein adsorption, monoclonal antibody binding, and platelet adhesion. AB - Ten specially synthesized polyurethanes (PUs) were used to investigate the effects of surface properties on platelet adhesion. Surface composition and hydrophilicity, fibrinogen (Fg) and von Willebrand's factor (vWf) adsorption, monoclonal anti-Fg binding, and platelet adhesion were measured. PUs preadsorbed with afibrinogenemic plasma or serum exhibited very low platelet adhesion, while adhesion after preadsorption with vWf deficient plasma was not reduced, showing that Fg is the key plasma protein mediating platelet adhesion under static conditions. Platelet adhesion to the ten PUs after plasma preadsorption varied greatly, but was only partially consistent with Fg adsorption. Thus, while very hydrophilic PU copolymers containing PEG that had ultralow Fg adsorption also had very low platelet adhesion, some of the more hydrophobic PUs had relatively high Fg adsorption but still exhibited lower platelet adhesion. To examine why some PUs with high Fg adsorption had lower platelet adhesion, three monoclonal antibodies (mAbs) that bind to sites in Fg thought to mediate platelet adhesion were used. The antibodies were: M1, specific to gamma-chain C-terminal; and R1 and R2, specific to RGD containing regions in the alpha-chain N- and C-terminal, respectively. Platelet adhesion was well correlated with M1 binding, but not with R1 or R2 binding. When these mAbs were incubated with plasma preadsorbed surfaces, they blocked adhesion to variable degrees. The ability of the R1 and R2 mAbs to partially block adhesion to adsorbed Fg suggests that RGD sites in the alpha chain may also be involved in mediating platelet adhesion and act synergistically with the C-terminal of the gamma-chain. PMID- 16037939 TI - Bioresorbable composites prepared by supercritical fluid foaming. AB - Bone is a natural composite construct, with a gradient structure going from a loose interconnected cellular core to an outer dense wall, thus minimizing bone weight while keeping a high mechanical resistance. Due to this unique and complex structure, bone defects are difficult to replace or repair. Tissue engineering aims at providing artificial bone grafts. Several techniques have been proposed to produce porous structures or scaffolds, but, as yet, with no optimal solutions. This article focuses on bioresorbable ceramic-polymer composite foams obtained by supercritical fluid foaming. This flexible technique enables an adequate morphology and suitable properties for bone tissue engineering to be obtained. Composite scaffolds are biocompatible, allowing cell proliferation and differentiation. PMID- 16037940 TI - Histological and radiographic evaluation of polymethylmethacrylate with two different concentrations of barium sulfate in a sheep vertebroplasty model. AB - Percutaneous vertebral augmentation with PMMA has been widely performed and usually provides good pain relief and stabilization of fractured vertebrae. Adequate visualization of PMMA during injection is desirable to minimize cement extravasation, so contrast agents such as barium sulfate are commonly added to the PMMA. The aim of this study was to evaluate the differences of histology and radiographic visualization when different concentrations of barium sulfate are mixed with PMMA. Six sheep were utilized in this study. Three vertebrae of each animal were exposed via retroperitoneal approach, and a cavity was created and then filled with either 10% or 30% BaSO4/PMMA, or left empty. Vertebrae were harvested and analyzed radiographically and histologically 12 and 90 days after surgery. Average CT value of the 30% BaSO4/PMMA group was 2.4-fold higher than that of the 10% BaSO4/PMMA group. Foreign-body giant cells were recognized around BaSO4particles at 90 days in the 30% BaSO4 group, whereas few particles were recognized in the 10% group at 90 days, or in either group at 12 days. A very mild giant-cell reaction is induced by a higher concentration of BaSO4 in PMMA, but the marked improvement in cement visualization by increased BaSO4 may be important to minimize more serious complications of cement extravasation during PMMA injection. PMID- 16037941 TI - Discerning alumina ball wear from confounding metal transfer artifact. AB - On dismounting a ceramic femoral ball from its metal trunnion, there is usually a range of gray metallic bands transferred to the trunnion bore inside the ball. This creates an artifact that may compromise detection by weight of the exceedingly low wear rates of ceramic balls. The objective of this study was to compare the weight trending of the metal trunnions and their ceramic balls during conditioning studies. Our hypothesis was that a pretest conditioning protocol would eliminate or greatly reduce the metal transfer artifact. The balls and tapers were placed on a hip simulator under 300-600 load cycles but with no articulation. The balls were then dismounted from the trunnions, and both were cleaned and weighed. This was repeated 6-23 times. We developed a novel hydraulic method for dismounting balls from trunnions that proved to be safe and efficient. There was significant weight loss in the trunnions after the ball removal, but there was no corresponding weight gain in the alumina balls. The weight effect of the metal transfer appeared to have been removed from the balls with our standard cleaning procedures. Therefore, wear rates for alumina balls may be gravimetrically determined without compensating for the metal transfer from trunnion to ball. PMID- 16037942 TI - Type IV collagen induces down-regulation of steroidogenic response to gonadotropins in adult rat Leydig cells involving mitogen-activated protein kinase. AB - We have previously shown that type IV collagen (alpha1 (IV) and alpha2 (IV) collagen chains) (Col-IV) inhibits testosterone (T) production by Leydig cells (LC). The aim of this study was to analyze mechanism/s by which Col-IV exerts this effect. No significant differences in the specific binding of hCG to LH/hCG receptors in LC cultured on uncoated or Col-IV coated plates were observed. An inhibition of cAMP production in hCG-stimulated LC cultured on Col-IV was detected. The inhibition exerted by Col-IV on T production in response to hCG was also observed when cells were stimulated with 8Bromo-cAMP. In addition, conversion of steroid precursors to T in LC cultured on uncoated and Col-IV coated plates was similar. On the other hand, we detected an increase of ERK1/2 phosphorylation in hCG-stimulated LC cultured on Col-IV. Genistein added to LC cultures reduced the ability of Col-IV to increase ERK1/2 phosphorylation and reverted the inhibitory effect of Col-IV on T production. An inhibitor of MEK, PD98059 added to LC cultures also reverted the inhibitory effect of Col-IV on T production. A decrease of steroidogenic acute regulatory protein (StAR) expression in hCG-stimulated LC cultured on Col-IV coated plates that could be reverted by addition of PD98059 to the cultures was also demonstrated. All together these results suggest that Col-IV inhibits T production in LC by binding to integrins, activating ERK1/2, decreasing cAMP production and decreasing StAR expression. PMID- 16037943 TI - VPAC1 expression is regulated by FXR agonists in the human gallbladder epithelium. AB - Vasoactive intestinal peptide receptor-1 (VPAC1) is the high-affinity receptor of vasoactive intestinal peptide (VIP), a major regulator of bile secretion. To better define the level at which VPAC1 stimulates bile secretion, we examined its expression in the different cell types participating in bile formation (i.e., hepatocytes, bile duct, and gallbladder epithelial cells). Because VPAC1 expression was previously shown to be regulated by nuclear receptors, we tested the hypothesis that it may be regulated by the farnesoid X receptor (FXR). Quantitative RT-PCR and immunoblot analyses of cell isolates indicated that VPAC1 is expressed in all three cell types lining the human biliary tree, with predominant expression in the gallbladder. In primary cultures of human gallbladder epithelial cells, VIP induced cAMP production and chloride secretion. Analysis of the VPAC1 gene revealed the presence of potential FXR response element sequences, and both FXR and RXRalpha expressions were detected in gallbladder epithelial cells. In these cells, the FXR pharmacological agonist GW4064 upregulated VPAC1 expression in a dose-dependent manner, and this effect was antagonized by the RXRalpha ligand, 9-cis retinoic acid. Chenodeoxycholate activated endogenous FXR in gallbladder epithelial cells, as ascertained by electromobility shift assay and upregulation of the FXR target gene, small heterodimer partner. Chenodeoxycholate also provoked an increase in VPAC1 mRNA and protein content in these cells. In conclusion, FXR agonists may increase gallbladder fluid secretion through transcriptional activation of VPAC1, which may contribute to the regulation of bile secretion by bile salts and to a protective effect of FXR pharmacological agonists in gallstone disease. PMID- 16037944 TI - Proteasome inhibition sensitizes hepatocellular carcinoma cells, but not human hepatocytes, to TRAIL. AB - TRAIL exhibits potent anti-tumor activity on systemic administration in mice. Because of its proven in vivo efficacy, TRAIL may serve as a novel anti neoplastic drug. However, approximately half of the tumor cell lines tested so far are TRAIL resistant, and potential toxic side effects of certain recombinant forms of TRAIL on human hepatocytes have been described. Pretreatment with the proteasome inhibitor MG132 and PS-341 rendered TRAIL-resistant hepatocellular carcinoma (HCC) cell lines but not primary human hepatocytes sensitive for TRAIL induced apoptosis. We investigated the different levels of possible MG132-induced interference with resistance to apoptotic signal transduction. Although proteasome inhibition efficiently suppressed nuclear factor-kappaB (NF-kappaB) activity, specific suppression of NF-kappaB by mutIkappaBalpha failed to sensitize TRAIL-resistant cell lines for TRAIL-induced apoptosis. In contrast to the previously reported mechanism of sensitization by 5-fluorouracil (5-FU), cellular FLICE-inhibitory protein (cFLIP)(L) and cFLIP(S) were markedly upregulated in the TRAIL death inducing signaling complex (DISC) by proteasome inhibitor pretreatment. Compared with 5-FU pretreatment, caspase-8 was more efficiently recruited to the DISC in MG132 pretreated cells despite the presence of fewer death receptors and more cFLIP in the DISC. But downregulation of cFLIP by short interference RNA (siRNA) further sensitized the HCC cell lines. In conclusion, these results show that otherwise chemotherapy-resistant tumor cells can be sensitized for TRAIL-induced apoptosis at the DISC level in the presence of high levels of cFLIP, which suggests the existence of an additional factor that modulates the interaction of FADD and the TRAIL death receptors. Of clinical relevance, proteasome inhibitors sensitize HCC cells but not primary human hepatocytes for TRAIL-induced apoptosis. PMID- 16037945 TI - LKM1 autoantibodies in chronic hepatitis C infection: a case of molecular mimicry? AB - Anti-liver-kidney microsome type 1 (LKM1) autoantibodies directed against the cytochrome P450 2D6 (CYP2D6) are considered specific markers of type 2 autoimmune hepatitis, but are also found in 5% of sera from patients chronically infected by hepatitis C virus (HCV). Molecular mimicry between HCV proteins and CYP2D6 has been proposed to explain the emergence of these autoantibodies. Anti-LKM1 autoantibodies from hepatitis C-infected patients were affinity-purified against immobilized CYP2D6 protein and used to screen a phage display library. CYP2D6 conformational epitopes were identified using phage display analysis and the identification of statistically significant pairs (SSPs). Cross-reactivity between CYP2D6 and HCV protein candidates was tested by immunoprecipitation. Nineteen different clones were isolated, and their sequencing resulted in the mapping of a conformational epitope to the region of amino acids 254-288 of CYP2D6. Candidate HCV proteins for molecular mimicry included: core, E2, NS3 and NS5a. Affinity-purified autoantibodies from HCV+/LKM1+ patients immunoprecipitated either NS3, NS5a, or both, and these reactivities were specifically inhibited by immobilized CYP2D6. In conclusion, HCV+/LKM1+ sera recognize a specific conformational epitope on CYP2D6 between amino acids 254 to 288, the region that contains the major linear epitope in type 2 autoimmune hepatitis patients. Cross-reactivity due to molecular mimicry at the B-cell level was shown between the CYP2D6 and the HCV NS3 and NS5a proteins and could explain the presence of anti-LKM1 in patients chronically infected with HCV. Further investigation of the role played by this molecular mimicry in HCV-infected patients may lead to more specific strategies for diagnosis and treatment. PMID- 16037946 TI - Spectrum of NAFLD and diagnostic implications of the proposed new normal range for serum ALT in obese women. AB - The upper limit of normal for ALT activity has been recommended to be lowered to < or = 30 U/L in men and < or = 19 U/L in women. These changes have been suggested to be diagnostically useful in subjects with nonalcoholic fatty liver disease (NAFLD). Our aim was to investigate the prevalence and spectrum of NAFLD with regard to the new ALT guidelines in 233 women with class II/III obesity. We compared our prior reference range for ALT (ULN < or = 30 U/L in women) with the new standard. Our study demonstrates that only 86 patients (36.9%) would be classified as having normal ALT levels compared with 169 patients (72.5%) by the new and old standards, respectively. In patients with normal ALT activity (new vs. old standard), the prevalence of fatty liver (FL: 39.5% vs 40.2%), portal fibrosis, and steatosis (IPF: 37.2% vs. 33.7%) and nonalcoholic steatohepatitis (NASH: 23.3% vs. 26%) were similar. In comparison, newly defined patients with elevated ALT levels (>19 U/L) demonstrated an increased prevalence of FL (36%) and IPF (11.6%) but a 23.8% decrease in the prevalence of NASH as compared with the old standard. The sensitivity and specificity for NASH were 42% and 80% (ALT > 30 U/L) compared with 74% and 42% (ALT > 19 U/L). In conclusion, a significant increase in the prevalence of FL and IPF is detected in subjects with elevated ALT levels with the application of the new standard. However, the diagnostic utility for ALT to identify NASH or IPF remains poor, and significant healthcare expenditures may be incurred if this standard is adopted. PMID- 16037947 TI - Expanded mesencephalic precursors develop into grafts of densely packed dopaminergic neurons that reinnervate the surrounding striatum and induce functional responses in the striatal neurons. AB - The search for alternative sources of dopaminergic cells, other than primary fetal tissue for transplantation in Parkinson's disease has become a major focus of research. Different methodological approaches have led to generation in vitro of cells expressing DA-cell markers, although these cells are frequently unable to survive for a long time in vivo after transplantation and/or induce functional effects in the host brain. In the present study, we grafted cell aggregates treated with antibodies against fibroblast growth factor 4 into dopaminergic denervated striata in rats. Furthermore, we grafted cell suspensions from primary mesencephalic fetal tissue. Grafts from expanded precursors were able to survive (at least 3 months postgrafting) and most decreased the lesion-induced ipsiversive rotation. In addition, immunolabeling for tyrosine hydroxylase and/or Fos showed that the grafts reinnervated the surrounding striatal tissue with dopaminergic terminals, and induced the expression of Fos in the striatal neurons of the reinnervated area after administration of amphetamine to the host rat. The number of dopaminergic cells in grafts from expanded precursors inducing rotational recovery was usually lower (1,226+/-314) than that in grafts from primary fetal tissue (1,671+/-122), but they were more densely packed in grafts that were of smaller volume and did not have the characteristic central nondopaminergic area observed in grafts from primary fetal tissue. The results suggest that long-term survival and functional integration into the DA-denervated striatum can be achieved with grafts of expanded mesencephalic precursors. PMID- 16037948 TI - Dual synaptic sites of D(1)-dopaminergic regulation of ethanol sensitivity of NMDA receptors in nucleus accumbens. AB - Regulation of NMDAreceptor-mediated synaptic transmission onto accumbal medium spiny neurons (MSN) may constitute an important site in drug reward and reinforcement in mesolimbic structures. Previously, we reported that D(1)-like dopamine receptors activate a postsynaptic cAMP/PKA/DARPP-32 signaling cascade culminating in phosphorylation of SER897-NR1 subunits and a reduction in the sensitivity to ethanol of NMDA receptor-mediated synaptic transmission. Here, we use a detailed electrophysiological analysis of D(1)-like receptor regulation of the ethanol sensitivity of accumbal NMDA receptors (NMDARs) through recordings of quantal Sr(2+)-supported NMDA miniature synaptic currents (mEPSCs) in reduced Mg(2+) (0.6 mM) and report dual presynaptic and postsynaptic components of D(1) like regulation of ethanol sensitivity of NMDARs. Ethanol inhibited NMDA mEPSC amplitude and frequency in a dose-dependent manner (25-75 mM), indicating inhibitory effects on presynaptic and postsynaptic components NMDA receptor mediated synaptic transmission. The presynaptic inhibitory effect was corroborated by analysing the ratio of paired-pulse facilitation (PPF) of Ca(2+) supported NMDA EPSCs. Activation of D(1) receptors with the agonist, SKF 38393 (25 microM), reversed ethanol suppression of NMDA mEPSC frequency and amplitude. Furthermore, the Mg(2+)-dependent decay off-rate of NMDA mEPSCs was substantially reduced by ethanol in a manner strongly reversed by the D(1) agonist. D(1) receptor-mediated attenuation of both the presynaptic and postsynaptic actions of ethanol was completely blocked by a D(1) selective antagonist (SCH 23390). These data suggest that D(1)-like receptors modulate both the presynaptic and postsynaptic effects of ethanol on NMDA receptor-mediated synaptic transmission in nucleus accumbens (NAc) and that these interactions may contribute to ethanol induced neuroadaptation of the reward pathway. PMID- 16037949 TI - Gompertz kinetics model of fast chemical neurotransmission currents. AB - At a chemical synapse, transmitter molecules ejected from presynaptic terminal(s) bind reversibly with postsynaptic receptors and trigger an increase in channel conductance to specific ions. This paper describes a simple but accurate predictive model for the time course of the synaptic conductance transient, based on Gompertz kinetics. In the model, two simple exponential decay terms set the rates of development and decline of transmitter action. The first, r, triggering conductance activation, is surrogate for the decelerated rate of growth of conductance, G. The second, r', responsible for Y, deactivation of the conductance, is surrogate for the decelerated rate of decline of transmitter action. Therefore, the differential equation for the net conductance change, g, triggered by the transmitter is dg/dt=g(r-r'). The solution of that equation yields the product of G(t), representing activation, and Y(t), which defines the proportional decline (deactivation) of the current. The model fits, over their full-time course, published records of macroscopic ionic current associated with fast chemical transmission. The Gompertz model is a convenient and accurate method for routine analysis and comparison of records of synaptic current and putative transmitter time course. A Gompertz fit requiring only three independent rate constants plus initial current appears indistinguishable from a Markov fit using seven rate constants. PMID- 16037950 TI - Increased AMPA GluR1 receptor subunit labeling on the plasma membrane of dendrites in the basolateral amygdala of rats self-administering morphine. AB - Glutamate-dependent synaptic plasticity is emerging as an important neural substrate of addiction. These drug-dependent neural adaptations may occur within brain systems that mediate reward, emotion, and cognitive function such as the amygdala complex. Modification of glutamate receptor targeting may be a key mechanism mediating neural plasticity; however, evidence for alteration of amygdala AMPA receptor localization in response to drug self-administration is lacking. High-resolution immunogold electron microscopic immunocytochemistry was used to compare surface and intracellular labeling of the calcium sensitive AMPA GluR1 receptor subunit in the basolateral (BLA) and central (CeA) nuclei of the amygdala in rats self-administering escalating doses of morphine or saline. Morphine self-administration was associated with regionally diverse effects on dendritic GluR1 targeting in the BLA and CeA. In the BLA of morphine self administering animals, there was a significant increase in the proportion of immunogold particles for GluR1 on the plasma membrane of dendrites, particularly in association with extrasynaptic sites, which was most prominent in large (2-4 microm) profiles. In contrast, there were no significant differences in surface or intracellular immunogold labeling in the CeA between morphine self administering and control animals. In both amygdala regions, GluR1 and the micro opioid receptor, the major cellular target of morphine, were only infrequently colocalized. These results indicate that GluR1 targeting is a dynamic process that can be differentially affected in distinct amygdala regions in response to chronic self-administration of morphine. Homeostatic adaptations in the subcellular localization of calcium sensitive AMPA receptors within the BLA may be an important neural substrate for alterations in reward, autonomic function, and behavioral processes associated with opiate addiction. PMID- 16037951 TI - Changes in in vivo [(3)H]-Ro15-4513 binding induced by forced swimming in mice. AB - Mice were forced to swim for 5 min in water at a temperature of 12 degrees C (cold water swim stress) or 32 degrees C (warm water swim stress), and stress induced analgesia (SIA) was measured using the tail-flick test. The cold water swim stress induced non-opioid SIA as well as hypothermia, whereas the warm water swim stress caused opioid SIA. The in vivo binding of [(3)H]-Ro15-4513 was measured in the stressed mice and compared with that in control mice. The specific binding of [(3)H]-Ro15-4513 in the cerebral cortex, hippocampus, and cerebellum was significantly altered by forced swimming in cold water. Apparent association and dissociation rate of [(3)H]-Ro15-4513 binding were decreased, and the change in the dissociation rate was most pronounced in the hippocampus. In contrast, no significant alterations were observed in in vitro binding. The hypothermia induced by the cold water swim stress seems to be the main reason for alterations in the specific binding of [(3)H]-Ro15-4513. The kinetics of a saturable amount of [(3)H]-Ro15-4513 in the blood and brain were also measured. The relative ratio of the radioactivity concentration in the brain to that in the blood was significantly decreased by forced swimming in cold water, indicating that the cold water swim stress induced changes in the nonspecific binding of [(3)H]-Ro15-4513 in the brain. These results together with previous reports suggested that non-opioid SIA induced by the cold water swim stress might be related to alterations in the rates of general ligand-receptor interactions including GABA(A)/benzodiazepine system. Changes in the nonspecific binding might be also involved in non-opioid SIA. PMID- 16037952 TI - Possible role of mitochondria in posttetanic potentiation of GABAergic synaptic transmission in rat neocortical cell cultures. AB - It has been previously demonstrated that mitochondria are of crucial importance for posttetanic potentiation (PTP) at neuromuscular junction. The aim of our study was to examine whether this may also be the case at a central synapse. To address this question, we studied possible mitochondrial involvement in PTP of GABAergic synaptic transmission in rat neocortical cultures, a preparation in which PTP has not been previously documented. Synaptic responses were evoked by local extracellular stimulation. Whole-cell patch-clamp technique was employed to record inhibitory postsynaptic currents (IPSCs) from postsynaptic neurons. Tetanic stimulation (30 Hz, 4 s) of the presynaptic neuron evoked an increase of IPSC amplitude, lasting for about 1 min. PTP was accompanied by a decrease of coefficient of variation of the IPSC and a decrease of paired-pulse (IPSC(2)/IPSC(1)) ratio, indicating involvement of presynaptic mechanism(s) in PTP. Possible role of mitochondria in PTP was addressed using drugs affecting Ca(2+) uptake and subsequent Ca(2+) efflux: carbonyl cyanide 3 chlorophenylhydrazone (CCCP) and tetraphenylphosphonium ions (TPP(+)). It was found that both CCCP (1-2 microM) and TPP(+) (10 microM) either substantially decreased or eliminated PTP. These results further confirm presynaptic origin of PTP in neocortical neurons and suggest an important role of mitochondrial Ca(2+) turnover in this form of synaptic plasticity at the central synapse. PMID- 16037953 TI - Imparting mineral affinity to proteins with thiol-labile disulfide linkages. AB - Chemical conjugation of bisphosphonates (BPs) to proteins is an effective means to enhance binding of proteins to mineral-containing biomaterials. BPs linked to proteins with reversible (i.e., cleavable) linkages were considered desirable over the conjugates linked with stable linkages because cleavable linkages allow protein release in free form from the mineral-containing biomaterials. To explore the feasibility of creating cleavable BP-protein conjugates, an amine- and a thiol-containing BP were conjugated to the model protein Bovine Serum Albumin (BSA) with N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), which resulted in disulfide-linked BP-BSA conjugates. Although disulfide-linked conjugates were stable under aqueous conditions, the conjugates in solution were readily cleaved in the presence of physiological concentrations (approximately 0.3 mM) of the thiol compound, cysteine. The imparted mineral affinity as a result of BP conjugation, as assessed by hydroxyapatite (HA) binding in vitro, was lost upon cleavage of the disulfide-linked BP. The conjugates bound to HA were also cleavable with cysteine, but their cleavage rate was significantly reduced as compared to the conjugates in solution. In conclusion, disulfide-linked BP conjugates were shown to be readily cleavable by the amino acid cysteine and this resulted in the loss of imparted mineral affinity of the proteins. The proposed approach will be useful for modulating in vivo delivery of proteins implanted with mineral-containing biomaterials. PMID- 16037954 TI - Influence of near null magnetic field on in vitro growth of potato and wild Solanum species. AB - The influence of near null magnetic field on in vitro growth of different cultures of potato and related Solanum species was investigated for various exposure times and dates. Potato (Solanum tuberosum L. cv. Desiree) in vitro cultures of shoot tips or nodal segments were used. Three different exposure periods revealed either stimulation or inhibition of root, stem, or leaf in vitro growth after 14 or 28 days of exposure. In one experiment the significant stimulation of leaf growth was also demonstrated at biochemical level, the quantity of chlorophyll a and b and carotenoids increasing more than two-fold. For the wild species Solanum chacoense, S. microdontum, and S. verrucosum, standardized in vitro cultures of nodal stem segments were used. Root and stem growth was either stimulated or slightly inhibited after 9 days exposure to near null magnetic field. Callus cultures obtained from potato dihaploid line 120/19 were maintained in near null magnetic field in 2 different months. For these experiments as well as for Solanum verrucosum, callus cultures recorded either slight inhibition or no effect on fresh weight. For all experiments significant growth variation was brought about only when geomagnetic activity (AP index) showed variations at the beginning of in vitro growth and when the explant had at least one meristematic tissue. Moreover longer maintenance in near null magnetic field, 28 days as compared to 14 days or the controls, can also make a difference in plant growth in response to geomagnetic field variations when static component was reduced to zero value. These results of in vitro plant growth stimulation by variable component of geomagnetic field also sustain the so-called seasonal "window" effect. PMID- 16037955 TI - Evolution of the spermatozoon in muroid rodents. AB - In the rodent superfamily Muroidea, a model for the evolution of sperm form has been proposed in which it is suggested that a hook-shaped sperm head and long tail evolved from a more simple, nonhooked head and short tail in several different subfamilies. To test this model the shape of the sperm head, with particular emphasis on its apical region, and length of sperm tail were matched to a recent phylogeny based on the nucleotide sequence of several protein-coding nuclear genes from 3 families and 10 subfamilies of muroid rodents. Data from the two other myomorph superfamilies, the Dipodoidea and kangaroo rats in the Geomyoidea, were used for an outgroup comparison. In most species in all 10 muroid subfamilies, apart from in the Murinae, the sperm head has a long rostral hook largely composed of acrosomal material, although its length and cross sectional shape vary across the various subfamilies. Nevertheless, in a few species of various lineages a very different sperm morphology occurs in which an apical hook is lacking. In the outgroups the three species of dipodid rodents have a sperm head that lacks a hook, whereas in the heteromyids an acrosome containing apical hook is present. It is concluded that, as the hook-shaped sperm head and long sperm tail occur across the muroid subfamilies, as well as in the heteromyid rodents, it is likely to be the ancestral condition within each of the subfamilies with the various forms of nonhooked sperm heads, that are sometimes associated with short tails, being highly derived states. These findings thus argue against a repeated evolution in various muroid lineages of a complex, hook shaped sperm head and long sperm tail from a more simple, nonhooked sperm head and short tail. An alternative proposal for the evolution of sperm form within the Muroidea is presented in the light of these data. PMID- 16037956 TI - New experimental approach to study host tissue response to surgical mesh materials in vivo. AB - Implantation of surgical meshes is a common procedure to increase abdominal wall stability in hernia repair. To improve biocompatibility of the implants, sophisticated in vivo animal models are needed to study inflammation and incorporation of biomaterials. Herein, we have established a new model that allows for the quantitative analysis of host tissue response and vascular ingrowth into surgical mesh materials in vivo. Ultrapro meshes were implanted into dorsal skinfold chambers of Syrian golden hamsters. Angiogenesis, microhemodynamics, microvascular permeability, and leukocyte-endothelial cell interaction of the host tissue were analyzed in response to material implantation over a 2-week period using intravital fluorescence microscopy. Mesh implantation resulted in a short-term activation of leukocytes, reflected by leukocyte accumulation and adherence in postcapillary venules. This cellular inflammatory response was accompanied by an increase of macromolecular leakage, indicating loss of integrity of venular endothelial cells. Angiogenesis started at day 3 after implantation by protrusion of capillary sprouts, originating from the host microvasculature. Until day 10, these sprouts interconnected with each other to form a new microvascular network. At day 14, the inflammatory response had disappeared and the vascular ingrowth was completed. Histology confirmed the formation of granulation tissue with adequate incorporation of the mesh filaments within the host tissue. We conclude that this novel model of surgical mesh implantation is a useful experimental approach to analyze host tissue response and vascular ingrowth of newly devised materials for hernia repair. PMID- 16037957 TI - Effects of a strong static magnetic field on bacterium Shewanella oneidensis: an assessment by using whole genome microarray. AB - The effect of a strong static 14.1 T magnetic field on log phase cells of bacterial strain Shewanella oneidensis MR-1 was evaluated by using whole genome microarray of this bacterium. Although differences were not observed between the treatment and control by measuring the optical density (OD), colony forming unit (CFU), as well as post-exposure growth of cells, transcriptional expression levels of 65 genes were altered according to our microarray data. Among these genes, 21 were upregulated while other 44 were downregulated, compared with control. PMID- 16037958 TI - Effects of 900 MHz electromagnetic fields exposure on cochlear cells' functionality in rats: evaluation of distortion product otoacoustic emissions. AB - In recent years, the widespread use of mobile phones has been accompanied by public debate about possible adverse consequences on human health. The auditory system is a major target of exposure to electromagnetic fields (EMF) emitted by cellular telephones; the aim of this study was the evaluation of possible effects of cellular phone-like emissions on the functionality of rat's cochlea. Distortion Products OtoAcoustic Emission (DPOAE) amplitude was selected as cochlea's outer hair cells (OHC) status indicator. A number of protocols, including different frequencies (the lower ones in rat's cochlea sensitivity spectrum), intensities and periods of exposure, were used; tests were carried out before, during and after the period of treatment. No significant variation due to exposure to microwaves has been evidenced. PMID- 16037959 TI - Effects of a 50 Hz electric field on plasma lipid peroxide level and antioxidant activity in rats. AB - The effects of exposure to extremely low frequency electric fields (ELF EFs) on plasma lipid peroxide levels and antioxidant activity (AOA) in Sprague-Dawley rats were studied. The test was based on comparisons among rats treated with a combination of the oxidizing agent, 2,2'-azobis(2-aminopropane) dihydrochloride (AAPH) and 50 Hz EF of 17.5 kV/m intensity for 15 min per day for 7 days, AAPH alone, EF alone or no treatment. EF significantly decreased the plasma peroxide level in rats treated with AAPH, similar to treatment by ascorbic acid or the superoxide dismutase. Ascorbic acid increased AOA; however, EF and superoxide dismutase did not change AOA compared with sham exposure in stressed rats. No influence on the lipid peroxide level and AOA in unstressed rats was observed with EF exposure alone. Although the administration of AAPH decreased AOA, this decrease did not change when EF was added. These data indicate that the ELF EF used in this study influenced the lipid peroxide level in an oxidatively stressed rat. PMID- 16037960 TI - Contact current hypothesis: summary of results to date. AB - Research conducted over the past 5 years has addressed the hypothesis that the reported association between residential magnetic fields and childhood leukemia may be explained by exposure to contact current. The use of multi-grounded neutrals in electrical distribution and residential electrical wiring systems in the United States results in a voltage on a residence's water line relative to earth that in turn creates a voltage between the water fixtures of a bathtub, sink, or shower and the drain, if the latter is made of conductive material. A bathing child may thus be exposed to contact current upon manual contact with the faucet, spout, or water stream. Dosimetry modeling indicates that modest and realistically anticipated currents (10s of microA) can produce electric fields in bone marrow (100s of mV/m) sufficient to overcome questions of biophysical plausibility. Both measurements in two regions of the United States and computer modeling of typical single-residence US neighborhoods indicate that residences with average magnetic fields in the high tail of the magnetic field distribution are more likely than residences with lower fields to also have higher contact voltage. The association of residential magnetic fields with contact voltage, the dosimetry results, and the indication from a behavioral survey that children tend to engage in behavior that results in exposure all support the hypothesis. Further research is needed to characterize electrical systems in other nations to determine whether contact current exposure occurs and whether it is associated with residential magnetic fields. PMID- 16037961 TI - Developmental effects of electromagnetic fields. AB - This paper reviews experimental studies on the effects of radiofrequency (RF), extremely low frequency (ELF), and intermediate frequency (IF) electromagnetic fields on animal development. Numerous studies have shown that RF fields are teratogenic at exposure levels sufficiently high to cause significant increase of temperature. There is no consistent evidence of RF field effects at nonthermal exposure levels. Only a few studies have evaluated possible effects on postnatal development using sensitive endpoints, such as behavioral effects. ELF electric fields up to 150 kV/m have been evaluated in several mammalian species. The results are rather consistent and do not suggest adverse developmental effects. The results of studies on ELF magnetic fields suggest effects on bird embryo development, but not consistently in all studies. Results from experiments with other non-mammalian experimental models have also suggested subtle effects on developmental stability. In mammals, most studies have shown no effects of prenatal exposure to ELF or IF magnetic on gross external, visceral, or skeletal malformations. The only finding that shows some consistency is increase of minor skeleton alterations in several experiments. Taken as a whole, the results do not show robust adverse effects of ELF and IF fields on development. However, additional studies on the suggested subtle effects on developmental stability might increase our understanding of the sensitivity of biological organisms to weak low-frequency magnetic fields. PMID- 16037962 TI - Validation of a micro-CT technique for measuring volumetric wear in retrieved acetabular liners. AB - In this study, a novel micro-CT-based technique for evaluating wear in retrieved acetabular liners was introduced and validated. Six UHMWPE acetabular components ranging in implantation time from 2.7 to 14.4 years were collected and evaluated with the use of a high-resolution micro-CT scanner. The components were scanned with a uniform volumetric resolution of 74 microns (16-bit precision) with the use of a 1,024 x 1,024 in-plane image matrix. Manual rigid 3D image registration of the interior hemispherical portion of the acetabular cup with geometric primitives by trained observers allowed for isolation, visualization, and measurement of the wear volume. Results for these six components indicated an average wear rate of 65 mm(3)/year. Overall scanner error was quantified gravimetrically and associated with a maximum uncertainty of 0.6%. Intra-- and interobserver uncertainty analysis showed the method to be both accurate and repeatable. PMID- 16037963 TI - Thermogelling emulsions for vascular embolization and sustained release of drugs. AB - Thermogelling emulsion system was developed to function as an embolic agent and sustained release system. PEG-PLGA-PEG triblock copolymer was synthesized, and blended with oily phase (Lipiodol(R)) to constitute the thermogelling emulsions. Because the polymer-rich aqueous phase dramatically increases viscosity in response to temperature change, especially within the range between 20 and 30 degrees C, the emulsions produce a stop-flowing gel with oil droplets entrapped. Thereafter, paclitaxels were released from the oily reservoir of gelled emulsions in a controlled manner. Reduced burst effect and steady drug release with near zero-order release kinetics were observed. Human umbilical vein endothelial cells (HUVEC) were collected from fresh umbilical cords for in vitro antiangiogenesis test. It demonstrated that the sustained release of paclitaxel from emulsions inhibited growth of HUVEC and that the IC(50), calculated according to release rate, was consistent with that obtained from free drug study. In addition, the emulsions forming a depot in situ inside the injection site of the blood vessel in rabbit ear obstructed the blood flow, and being monitored under X-ray angiography. Taken together, this study proved the feasibility of the thermogelling emulsions for vascular embolization and sustained drug release. The results presented a potential system for arterial transcatheter embolization on hepatocellular carcinoma combined with anti-angiogeneic treatment. PMID- 16037964 TI - Degradation of poly-L/DL-lactide versus TCP composite pins: a three-year animal study. AB - Biodegradable polylactide implants allow secure fixation of osteochondral fractures. This quasirandomized parallel-group animal study investigates whether a composite implant of poly-L/DL-lactide (PLDLLA) with additional 10% beta tricalcium phosphate produces an osteoconductive effect, whether the admixture positively influences implant degradation (assessed by comparing time to implant degradation), and whether the pin sites of degraded implants are replaced with bone tissue. On 36 medial femoral condyles of sheep, osteotomies were fixed with either three PLDLLA pins or three composite pins. At 3, 18, and 36 months, the pin sites were measured histologically and the state of degradation assessed according to Pistner's classification. All fractures healed without clinically relevant complications and without displacement. Both pin types led to asymptomatic pin-site enlargement at 18 months, which disappeared by 36 months. At 18 months, 14 of 18 PLDLLA pins were no longer evident, whereas 16 of 18 composite pins showed only peripheral degradation. By 36 months, all implants from both groups were completely degraded and replaced with scar (2/18) or bone tissue (16/18). At 36 months there was no evidence for significant improvement of either degradation performance or osseointegration through use of the 10% TCP composite mixture. PMID- 16037965 TI - Physiologically clotted fibrin-calcined bone composite--a possible bone graft substitute. AB - Fibrin plays a major role in blood clotting and is important in wound healing. In the present study, physiologically clotted fibrin (PF) and calcined bone (CB) both of bovine origin were used in the preparation of a bone graft material with chitosan (C) as a binder. This bone graft (PF-CB) was characterized by stress strain analysis, X-ray diffraction (XRD) studies, scanning electron microscopy, differential scanning calorimetry, thermogravimetric analysis, and FTIR spectroscopy. Thermal studies revealed that the incorporation of CB into PF did not alter the thermal stability of fibrin, although it affects the total loss percentage. The composition containing 24:1:15 (PF:C:CB) on dry weight basis exhibited maximum compressive strength, and the same composite was used for characterization studies. The XRD studies have shown the amorphous nature of the implant. SEM pictures revealed the heterogeneity and porous nature of the implant. PF in combination with CB is expected to exhibit osteoinductivity because fibrin, the osteoinductive protein, acts as an angiogenic factor, whereas CB provides calcium and phosphate ions that are needed for new bone formation. PMID- 16037966 TI - Injectable gels of anionic collagen:rhamsan composites for plastic correction: preparation, characterization, and rheological properties. AB - The present article describes the preparation and characterization of anionic collagen gels obtained from porcine intestinal submucosa after 72 h of alkaline treatment and in the form of rhamsan composites to develop injectable biomaterials for plastic reconstruction. All materials were characterized by SDS/polyacrylamide gel electrophoresis, infrared spectroscopy, thermal stability, potentiometric titration, rheological properties, and fluidity tests. Biocompatibility was appraised after the injection of anionic collagen: rhamsan composites at 2.5% in 60 North Folk rabbits. Independently of processing, the collagen's secondary structure was preserved in all cases, and after 72 h of hydrolysis the collagen was characterized by a carboxyl group content of 346+/-9, which, at physiological pH, corresponds to an increase of 106+/-17 negative charges, in comparison to native collagen, due to the selective hydrolysis of asparagine and glutamine carboxyamide side chain. Rheological studies of composites at pH 7.4 in concentrations of 2, 4, and 6% (in proportions of 75:1 and 50:1) showed a viscoelastic behavior dependent on the frequency, which is independent of concentration and proportion. In both, the concentration of the storage modulus always predominated over the loss modulus (G'>G'' and delta<45 degrees ). The results from creep experiments confirmed this behavior and showed that anionic collagen:rhamsan composites at pH 7.4 in the proportion of 50:1 are less elastic and more susceptible to deformation in comparison to gels in the proportion of 75:1, independent of concentration. This was further confirmed by flow experiments, indicating that the necessary force for the extrusion of anionic collagen:rhamsan composites, in comparison to anionic collagen, was significantly smaller and with a smooth flow. Biocompatibility studies showed that the tissue reaction of anionic collagen:rhamsan composites at 2.5% in the proportion of 75:1 was compatible with the application of these gels in plastic reconstruction. These results suggest that the association of collagen with rhamsan may be a good alternative in the replacement of glutaraldehyde to stabilize the microfibril assembly of commercial collagen gel preparations. PMID- 16037967 TI - Successful treatment of oral verrucous hyperplasia and oral leukoplakia with topical 5-aminolevulinic acid-mediated photodynamic therapy. AB - BACKGROUND AND OBJECTIVES: Our recent studies showed that a new topical 5 aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) protocol using a light-emitting diode (LED) light source is an effective and successful treatment modality for five cases of oral verrucous hyperplasia (OVH) and one case of verrucous carcinoma. In this study, we treated eight OVH lesions with the same topical ALA-PDT protocol to further confirm the efficacy of this protocol on OVH lesions. In addition, our recent study demonstrated an unsatisfactory clinical outcome for 24 oral leukoplakia (OL) lesions treated once a week by the same topical ALA-PDT protocol. Therefore, in this study 24 OL lesions were treated twice a week by the same protocol to compare whether the twice-a-week treatment modality could result in a better clinical outcome than the once-a-week treatment modality. STUDY DESIGN/MATERIALS AND METHODS: In this study, 8 OVH and 24 OL lesions were treated by the same topical ALA-PDT once a week and twice a week, respectively. RESULTS: All the former eight OVH lesions treated once a week showed complete response (CR) after 2-5 (mean, 3.8) treatments of ALA-PDT. The latter 24 OL lesions treated twice a week demonstrated CR in 8, partial response (PR) in 16, and no response in none. The present 24 OL lesions treated twice a week had a significantly better clinical outcome than the previous 24 OL lesions treated once a week (P = 0.000, chi-square test). CONCLUSIONS: We conclude that complete regression of OVH lesions can be achieved by less than six treatments of topical ALA-PDT once a week. Although the response of OL lesions to the topical ALA-PDT is not as good as the response of OVH lesions to the same therapy, all OL lesions can have at least PR after eight treatments with the topical ALA-PDT twice a week. In addition, OL lesions treated twice a week have a significantly better clinical outcome than OL lesions treated once a week. PMID- 16037968 TI - Evaluation of a novel flash lamp system (FLS) incorporating optimal spectral filtration for the treatment of photoaging. AB - BACKGROUND AND OBJECTIVES: Improvement in photoaging using laser and Intense Pulsed Light (IPL) modalities is well documented in the literature. We report a prospective study evaluating the safety and efficacy of a novel flash lamp IPL system incorporating a spectral filtration system designed to maximize improvement of facial dyschromias, telangiectases, and skin texture. The device was a prototype xenon flashlamp pulsed light. The novel features of this IPL device are the extended pulse duration and smooth temporal pulse profile. STUDY DESIGN/MATERIALS AND METHODS: We enrolled 23 patients (22 female, 1 male) of Fitzpatrick skin type I-IV with evidence of photoaging (lentigenes, ephelides, telangiectases, rhytides). Each patient underwent test sites on their back to establish safe treatment parameters. Using treatment fluences of 20-37 J/cm(2), each patient was given 2-4 full-face treatments separated by 3-4 weeks. RESULTS: Test sites revealed that shorter wavelength filters and shorter pulse durations increased the risk of epidermal injury. Objective assessment by evaluation of pre and post treatment photographs of the face showed an average improvement of 53% in hyperpigmentation, 39% in telangiectases, and 8% in wrinkles. There were no significant adverse effects. CONCLUSIONS: The novel flash lamp IPL system in this study was safe and effective in reducing vascular and pigment dyschromias in patients with skin types I through IV. PMID- 16037969 TI - Assessing microleakage of class V resin composite restorations after Er:YAG laser and bur preparation. AB - BACKGROUND AND OBJECTIVE: This study aimed to evaluate the extent of microleakage in cavities prepared with bur and Er:YAG laser, hybridized with different bonding systems. STUDY DESIGN: Sixty bovine teeth were randomly divided into six groups (n = 10): (G1) Diamond bur + Single Bond; (G2) Diamond bur + AdheSE; (G3) Diamond bur + Clearfil SE Bond; (G4) Er:YAG (250 mJ, 4 Hz, 80.6 J/cm(2)) + Single Bond; (G5)Er:YAG + AdheSE, and (G6) Er:YAG + Clearfil. Cavities were restored with a micro-hybrid composite resin. After thermocycling, the specimens were stained with 2% methylene blue solution and sectioned in the mesiodistal direction. Dye penetration was scored based upon the extent of the dye using a light stereoscope. RESULTS: The Kruskal-Wallis and Mann-Whitney test revealed no statistically significant differences between the method of preparation (diamond bur and laser). However, statistical differences were found between the adhesives tested. CONCLUSION: Based on the results of this study, Er:YAG laser confirmed to be as effective as the conventional methods for preparing adhesive restorations. PMID- 16037970 TI - Modeling and simulations of the pharmacokinetics of fluorophore conjugated antibodies in tumor vicinity for the optimization of fluorescence-based optical imaging. AB - BACKGROUND AND OBJECTIVES: One of the methods to detect and localize tumors in tissue is to use fluorophore conjugated specific antibodies as tumor surface markers. The goals of this study are to understand and quantify the pharmacokinetics of fluorophore conjugated antibodies in the vicinity of a tumor. This study concludes another stage of the development of a non-invasive fluorescenated antibody-based technique for imaging and localization of tumors in vivo. STUDY DESIGN/MATERIALS AND METHODS: A mathematical model of the pharmacokinetics of fluorophore conjugated antibodies in the vicinity of a tumor was developed based on histological staining experiments. We present the model equations of concentrations of antibodies and free binding sites. We also present a powerful simulation tool that we developed to simulate the imaging process. We analyzed the model and studied the effects of various independent parameters on the imaging result. These parameters included initial volume of markers (injected volume), total number of binding sites, tumor size, binding and dissociation rate constants, and the diffusion coefficient. We present the relations needed between these parameters in order to optimize the imaging results. RESULTS AND CONCLUSIONS: A powerful and accurate tool was developed which may assist in optimizing the imaging system results by setting the injection volume and concentration of fluorophore conjugated antibodies in tissue and approximating the time interval where maximum specific binding occurs and the tumor can be imaged. PMID- 16037971 TI - Low power laser irradiation alters gene expression of olfactory ensheathing cells in vitro. AB - BACKGROUND AND OBJECTIVES: Both photobiomodulation (PBM) and olfactory ensheathing cells (OECs) transplantation improve recovery following spinal cord injury. However, neither the combination of these two therapies nor the effect of light on OECs has been reported. The purpose of this study was to determine the effect of light on OEC activity in vitro. MATERIALS AND METHODS: OECs were purified from adult rat olfactory bulbs and exposed to 810 nm light (150 mW; 0, 0.2, or 68 J/cm(2)). After 7-21 days in vitro, cells underwent immunocytochemistry or RNA extraction and RT-PCR. RESULTS: Analysis of immunolabeling revealed a significant decrease in fibronectin expression in the cultures receiving 68 J/cm(2). Analysis of gene expression revealed a significant (P < 0.05) increase in brain derived neurotrophic factor (BDNF), glial derived neurotrophic factor (GDNF), and collagen expression in the 0.2 J/cm(2) group in comparison to the non-irradiated and 68 J/cm(2) groups. OEC proliferation was also found to significantly increase in both light treated groups in comparison to the control group (P < 0.001). CONCLUSIONS: These results demonstrate that low and high dosages of PBM alter OEC activity, including upregulation of a number of neurotrophic growth factors and extracellular matrix proteins known to support neurite outgrowth. Therefore, the application of PBM in conjunction with OEC transplantation warrants consideration as a potential combination therapy for spinal cord injury. PMID- 16037972 TI - Surgical outcome and prognostic factors of cryptogenic neocortical epilepsy. AB - Surgical treatment of cryptogenic neocortical epilepsy is challenging. The aim of this study was to evaluate surgical outcomes and to identify possible prognostic factors including the results of various diagnostic tools. Eighty-nine patients with neocortical epilepsy with normal magnetic resonance imaging (35 patients with frontal lobe epilepsy, 31 with neocortical temporal lobe epilepsy, 11 with occipital lobe epilepsy, 11 with parietal lobe epilepsy, and 1 with multifocal epilepsy) underwent invasive study and focal surgical resection. Patients were observed for at least 2 years after surgery. The localizing values of interictal electroencephalogram (EEG), ictal scalp EEG, interictal 18F-fluorodeoxyglucose positron emission tomography (FDG-PET), and subtraction ictal single-photon emission computed tomography were evaluated. Seventy-one patients (80.0%) had a good surgical outcome (Engel class 1-3); 42 patients were seizure free. Diagnostic sensitivities of interictal EEG, ictal scalp EEG, FDG-PET, and subtraction ictal single-photon emission computed tomography were 37.1%, 70.8%, 44.3%, and 41.1%, respectively. Localization by FDG-PET and interictal EEG was correlated with a seizure-free outcome. The localizing value of FDG-PET was greatest in neocortical temporal lobe epilepsy. The focalization of ictal onset and also ictal onset frequency in invasive studies were not related to surgical outcome. Concordance with two or more presurgical evaluations was significantly related to a seizure-free outcome. PMID- 16037973 TI - Transcranial ultrasound shows nigral hypoechogenicity in restless legs syndrome. AB - In patients with Parkinson's disease, hyperechogenicity of the substantia nigra using transcranial ultrasound has been related to increased tissue concentrations of iron. Recently, deficient iron transport mechanisms in substantia nigra neurons have been described in postmortem tissue of patients with restless legs syndrome (RLS). This study was performed to study substantia nigra echogenicity in RLS patients compared with normal control subjects and Parkinson's disease patients. RLS patients had significantly reduced midbrain areas of hyperechogenicity compared with control subjects, and even more markedly reduced hyperechogenicity compared with Parkinson's disease patients. These findings lend further support to nigral iron deficiency as a pathogenetic factor in RLS. PMID- 16037974 TI - Mutations in phenotypically mild D-2-hydroxyglutaric aciduria. AB - D-2-hydroxyglutaric aciduria is a neurometabolic disorder with mild and severe phenotypes. Recently, we reported pathogenic mutations in the D-2 hydroxyglutarate dehydrogenase gene as the cause of the severe phenotype of D-2 hydroxyglutaric aciduria in two patients. Here, we report two novel pathogenic mutations in this gene in one patient with a mild presentation and two asymptomatic siblings with D-2-hydroxyglutaric aciduria from two unrelated consanguineous Palestinian families: a splice error (IVS4-2A-->G) and a missense mutation (c.1315A-->G;p.Asn439Asp). Overexpression of this mutant protein showed marked reduction of the enzyme activity. PMID- 16037975 TI - Clinical course in young patients with sporadic Creutzfeldt-Jakob disease. AB - Sporadic Creutzfeldt-Jakob disease (sCJD) is a rare neurodegenerative disease with the greatest incidence occurring in patients between 60 and 70 years old. Younger patients may also be affected. In this study, we used all case material available from 52 patients with sCJD aged 50 years or younger at disease onset, who were identified between 1993 and 2003 in Germany. The objective of this study was to describe the psychiatric and neurological features of these young patients with emphasis on the different codon 129 genotypes and PrP types, and to compare them with elder patients with sCJD and patients with variant CJD. We also gave particular attention to electroencephalogram, magnetic resonance imaging, and 14 3-3 results, as well as to the neuropathological lesion profile. The clinical syndrome in young patients differs from elder patients with CJD with respect to clinical signs, disease duration, technical investigations, and neuropathological lesion profile. The psychiatric symptoms in young patients with sCJD are similar to the psychiatric symptoms expressed by patients with variant CJD; however, in contrast with the variant cases, young patients with sCJD experience development of prominent dementia early in the disease course. PMID- 16037976 TI - Elevation of 12/15 lipoxygenase products in AD and mild cognitive impairment. AB - The 12/15 lipoxygenase (12/15LOX) enzyme is increased in pathologically affected frontal and temporal regions of Alzheimer's disease (AD) brains compared with controls. Herein, we measured 12(S)-HETE and 15(S)-HETE levels, products of 12/15LOX, in cerebrospinal fluid (CSF) of normal individuals, subjects with mild cognitive impairment (MCI) and AD. Compared with controls, there was a significant increase of both metabolites in CSF from AD and MCI, which correlated with lipid peroxidation and tau protein levels. These results suggest that the activation of this enzyme occurs early in the course of AD, before the onset of overt dementia, thereby implicating 12/15LOX-mediated lipid peroxidation in the pathogenesis of AD. PMID- 16037977 TI - Electron microscopic radioautographic study of RNA synthesis in hepatocyte mitochondria of aging mouse. AB - In order to study the aging changes of intramitochondrial RNA synthesis in mouse hepatocytes, 10 groups of aging mice, each consisting of three individuals (total 30) from fetal day 19 to postnatal month 24 were injected with 3H-uridine, an RNA precursor, sacrificed 1 hour later, and the liver tissues processed for electron microscopic radioautography. On EM radioautograms obtained from each animal the number of mitochondria, the number of labeled mitochondria, and the mitochondrial labeling index labeled with 3H-uridine showing RNA synthesis in each hepatocytes, both mononucleate and binucleate cells, were counted and the averages in respective aging groups were compared. From the results it was demonstrated that the numbers of mitochondria, the numbers of labeled mitochondria, and the labeling indices of intramitochondrial RNA syntheses in both mononucleate and binucleate hepatocytes of mice at various ages increased and decreased according to the age of the animals. PMID- 16037978 TI - Experimental LPS-induced cholestasis alters subcellular distribution and affects colocalization of Mrp2 and Bsep proteins: a quantitative colocalization study. AB - Quantitative colocalization analysis is a powerful tool for reliable estimation of the colocalization of antigens. We employed it to determine the changes of colocalization of multidrug resistance protein 2 (Mrp2) and bile salt export pump (Bsep) in confocal immunofluorescence microscopy images of rat liver following lipopolysaccharide (LPS) administration. Samples were taken 2, 24, 48 hours, and 1 week after LPS challenge. Pearson's correlation coefficient (PCC), an overlap coefficient according to Manders' (MOC), and overlap coefficients k1 and k2 were used to explore changes of the degree of colocalization. In intact animals, confocal microscopy showed tight colocalization of Mrp2 and Bsep proteins exclusively at the bile canaliculi. High degree of colocalization was confirmed quantitatively. Injection of LPS resulted in the appearance of fuzzy-looking areas of fluorescence of both proteins around bile canaliculi 2 and 24 hours after administration and relocation of Mrp2 protein to the basolateral domain of hepatocytes at 48 hours. By 1 week, canalicular localization was restored morphologically. Quantitative colocalization analysis of canalicular regions showed a steady decrease of the degree of colocalization of Mrp2 and Bsep up to 48 hours with the slight increase of its value by 1 week. These findings demonstrate that Mrp2, in contrast to Bsep, is partially and reversibly relocated from canalicular to basolateral domain of hepatocytes after LPS challenge. PMID- 16037979 TI - Cilia in the head of hornets: form and function. AB - In the head of the Oriental hornet, beneath the cuticle, there are plaques of hair cells. These are distributed throughout the upper front part of the head; to wit: in the region of the vertex (i.e., around and behind the ocelli), in the genae around and behind the compound eyes (the ommatidia), and in the region of the forehead or frons. These hair cells are arranged with their thin whip-like part (i.e., cilia) directed outward and morphologically fall into three distinct groups: type (a) thin elongated cilia connected to each other alongside by side links; type (b) thin elongated cilia of which two or more interconnect at their distal ends via a delicate nerve fiber bearing a knob at its center; and type (c) shorter and thicker cilia that roughly resemble a triangular thorn and are also interconnected by a thin thread, which, however, bears a ball rather then a knob at its center. The knob in the one case and the ball in the other vary in their diameter, but in both instances the interconnecting elements, be they nerve fibers or threads, are seemingly multidirectional. Beneath the frons, in the region of the coronal suture, the hair cells (cilial plaques) are inwardly directed and bear a large trachea at their center. Presumably, the "weighted" cilial cells that are directed toward the exterior of the body aid the hornet in navigation and gravity determination whereas the inwardly directed ciliary cells may possibly serve in acoustic communication. Another element worthy of mention within the hair cells are yellow granules (yg). These yg's originate from the whip-like portion of the ciliary cells that are distributed beneath the frons plate, and also in the yellow stripes of the gastral cuticle. Conceivably, these yellow granules, in both cases, may play a role in the absorption and storage of solar energy. In summary, ciliary structures are involved in the hornet in gravity sensing, in acoustical communication and in light sensing, i.e., with some similarity with what happens in vertebrates in the inner ear and in the photoreceptor. PMID- 16037980 TI - Fluorescent human lung macrophages analyzed by spectral confocal laser scanning microscopy and multispectral cytometry. AB - Numerous highly fluorescent macrophages (MPhi), designated "smoker cells," exist in the lungs of smokers and subjects who have quit smoking within 5 years. The brightly fluorescent MPhi, however, are not present in the lungs of never smokers. Some investigators have speculated that the intense fluorescence of the MPhi is due to smoke-induced changes in the autofluorescence of naturally occurring (i.e., endogenous) compounds (e.g., NADP). In contrast, other researchers have theorized that the fluorescence is due to the uptake of tobacco smoke particulates (i.e., "tar"). Studies reported herein were undertaken to test the hypothesis that the origin of the MPhi fluorescence could be profiled with the novel technologies afforded by spectral confocal laser scanning microscopy (sCLSM) and multispectral cytometry (MSC). To this end, spectral emissions were obtained by sCLSM of optical sections of live MPhi isolated from fresh surgically excised human lung tissue and in air-dried lung tissue imprints. Confirmation of spectral profiles of these single cell observations was obtained in population studies with the use of high-throughput MSC in which multispectral analyses were performed with three different lasers. Proof of concept experiments demonstrated that relatively nonfluorescent MPhi from the lungs of nonsmokers became fluorescent upon short-term ex vivo exposure to tobacco smoke tar. Summarily, the studies reported herein document that the fluorescence of human lung MPhi is due to tobacco tar. PMID- 16037981 TI - Receptor binding occurrence and plasma levels of natriuretic peptides in response to sympathectomy. AB - In the present investigation the relationship between the sympathetic nervous system and the endocardial levels of receptor binding sites for natriuretic peptides and the plasma content of atrial natriuretic peptide were analyzed in rats. In order to destruct the cardiac sympathetic nerve terminals, chemical sympathectomy with 6-hydroxydopamine was made in parallel with intravenous measurements of blood pressure and heart frequency. By use of immunohistochemical and enzyme-linked-immunosorbent techniques the expression of tyrosine hydroxylase positive sympathetic nerve terminals and plasma levels of pro-atrial natriuretic peptide were determined, respectively. The occurrence of receptor binding sites for natriuretic peptides was examined by in vitro receptor autoradiography. In contrast to the marked occurrence of natriuretic peptide receptor binding sites seen in the ventricular endocardium of control rats, the sympathectomized rats exhibited a decreased number of binding sites for natriuretic peptides in the endocardium of both the right and left chambers. Interestingly, this was found in parallel with a significant decrease of systolic and diastolic blood pressure and increased plasma levels of pro-atrial natriuretic peptide in the treated group of rats. These findings, together with those in previous studies, give support to an idea that one part of the blood pressure-decreasing effects, seen in patients treated with beta-adrenergic blockade, might be through a reduction of the natriuretic clearance receptor C, then giving rise to increased levels of atrial natriuretic peptide. PMID- 16037982 TI - Growth model for arc-deposited fullerene-like CNx nanoparticles. AB - Multiwall CNx nanotubes, nanoonions, and amorphous nanoballs were formed by carbon DC arc evaporation in a nitrogen atmosphere. The samples were investigated by conventional and high-resolution transmission electron microscopy. We propose a fragment-by-fragment growth mechanism for the formation of the nanoparticles. Accordingly, particles and aggregates of particles form in the vacuum ambient by the collisions between atomic species and small fragments. This growth model is supported by the discontinuous inner shells and disordered surface layers composed from graphene fragments. Image simulations confirm the detectability of dangling and back-folding surface layers in the experimental images. Further, the simulated images also confirm that the growth of nanoonions starts from a single fullerene-like seed. The amorphous nanoballs form when ordering of the building blocks during growth is hindered by the cross-linking nitrogen bonds. PMID- 16037983 TI - Digital position determination system for electron microscopy. AB - The precise determination of object positions within a specimen grid is important for many applications in electron microscopy. For example, real-time position determination is necessary for current statistical approaches and the efficient mapping and relocation of objects. Unfortunately, precise real-time position determination is not available on many older electron microscopes with manual stage controls. This report demonstrates the cost-effective and flexible implementation of a digital position determination system that can be adapted to many hand-operated electron microscopes. A customized solution that includes the hardware and software to accomplish position determination is presented. Lists of required parts, instructions for building the hardware, and descriptions of the developed programs are included. Two LED-photodiode assemblies detect x and y movements via an optical wheel that is in physical contact with the mechanical x and y stage control elements. These detector assemblies are interfaced with an integrated circuit that converts movement information into serial port-compatible signals, which are interpreted by a computer with specialized software. Two electron microscopes, a Philips CM12 (S)TEM and a Philips 201 TEM, were equipped with the described digital position determination system. The position fidelity and position fidelity after reloading of grids were determined for both microscopes. The determined position deviation was 1.06 microm in the x axis and 0.565 microm in the y axis for the Philips CM12 (S)TEM, and 0.303 microm in the x axis and 0.545 microm in the y axis for the Philips 201 TEM. After reloading and computational realigning, the determined average position variation was 2.66 microm in the x axis and 2.61 microm in the y axis for the Philips CM12 (S)TEM, and 1.13 microm in the x axis and 1.27 microm in the y axis for the Philips 201 TEM. PMID- 16037984 TI - Generic head models for atlas-based EEG source analysis. AB - We describe a method for using a generic head model, in the form of an anatomical atlas, to produce EEG source localizations. The atlas is fitted to the subject by a nonrigid warp using a set of surface landmarks. The warped atlas is used to compute a finite element model (FEM) of the forward mapping or lead-fields between neural current generators and the EEG electrodes. These lead-fields are used to localize current sources from the subject's EEG data and the sources are then mapped back to the anatomical atlas. This approach provides a mechanism for comparing source localizations across subjects in an atlas-based coordinate system, which can be used in the large fraction of EEG studies in which MR images are not available. The Montreal brain atlas was used as the reference anatomical atlas and 10 individual MR volumes were used to evaluate the method. The atlas was fitted to each subject's head by a thin-plate-spline (TPS) warp. The spatial locations of a generic 155-electrode configuration were used to constrain the warp. For the purposes of evaluation, dipolar sources were placed on the inner cortical surface in the atlas geometry and transferred to each subject's brain space using a polynomial warp. The parameters of the warp were computed using an intensity-based matching of the atlas and subject brains, thus ensuring that the sources were placed at approximately the same anatomical location in each case. Data were simulated in the subject geometry and a dipole fit was performed on these data using an FEM of the TPS warped atlas. The source positions found in the warped atlas were transferred back to the original atlas and compared to the original position. Sources were simulated at 972 locations evenly distributed over the inner cortical surface of the atlas. The mean error over all 10 subjects was 8.1 mm in the subject space and 15.2 mm in the atlas space. In comparison, using an affine transformation of the electrodes into atlas space and an FEM model generated from the atlas produced mean errors of 22.3 mm in subject space and 19.6 mm in atlas space. With a standard three-shell spherical model the errors were 27.2 mm in the subject space and 34.7 mm when mapped to atlas space. PMID- 16037985 TI - Spatiotemporal mapping of cortical activity accompanying voluntary movements using an event-related beamforming approach. AB - We describe a novel spatial filtering approach to the localization of cortical activity accompanying voluntary movements. The synthetic aperture magnetometry (SAM) minimum-variance beamformer algorithm was used to compute spatial filters three-dimensionally over the entire brain from single trial neuromagnetic recordings of subjects performing self-paced index finger movements. Images of instantaneous source power ("event-related SAM") computed at selected latencies revealed activation of multiple cortical motor areas prior to and following left and right index finger movements in individual subjects, even in the presence of low-frequency noise (e.g., eye movements). A slow premovement motor field (MF) reaching maximal amplitude approximately 50 ms prior to movement onset was localized to the hand area of contralateral precentral gyrus, followed by activity in the contralateral postcentral gyrus at 40 ms, corresponding to the first movement-evoked field (MEFI). A novel finding was a second activation of the precentral gyrus at a latency of approximately 150 ms, corresponding to the second movement-evoked field (MEFII). Group averaging of spatially normalized images indicated additional premovement activity in the ipsilateral precentral gyrus and the left inferior parietal cortex for both left and right finger movements. Weaker activations were also observed in bilateral premotor areas and the supplementary motor area. These results show that event-related beamforming provides a robust method for studying complex patterns of time-locked cortical activity accompanying voluntary movements, and offers a new approach for the localization of multiple cortical sources derived from neuromagnetic recordings in single subject and group data. PMID- 16037986 TI - An analysis of variability in the manufacturing of dexosomes: implications for development of an autologous therapy. AB - Dexosomes are nanometer-size vesicles released by dendritic-cells, possessing much of the cellular machinery required to stimulate an immune response (i.e. MHC Class I and II). The ability of patient-derived dexosomes loaded with tumor antigens to elicit anti-tumor activity is currently being evaluated in clinical trials. Unlike conventional biologics, where variability between lots of product arises mostly from the manufacturing process, an autologous product has inherent variability in the starting material due to heterogeneity in the human population. In an effort to assess the variability arising from the dexosome manufacturing process versus the human starting material, 144 dexosome preparations from normal donors (111) and cancer patients (33) from two Phase I clinical trials were analyzed. A large variability in the quantity of dexosomes (measured as the number of MHC Class II molecules) produced between individual lots was observed ( > 50-fold). An analysis of intra-lot variability shows that the manufacturing process introduces relatively little of this variability. To identify the source(s) of variability arising from the human starting material, distributions of the key parameters involved in dexosome production were established, and a model created. Computer simulations using this model were performed, and compared to the actual data observed. The main conclusion from these simulations is that the number of cells collected per individual and the productivity of these cells of are the principal sources of variability in the production of Class II. The approach described here can be extended to other autologous therapies in general to evaluate control of manufacturing processes. Moreover, this analysis of process variability is directly applicable to production at a commercial scale, since the large scale manufacture of autologous products entails an exact process replication rather than scale-up in volume, as is the case with traditional drugs or biologics. PMID- 16037987 TI - Biotransformation of R-2-hydroxy-4-phenylbutyric acid by D-lactate dehydrogenase and Candida boidinii cells containing formate dehydrogenase coimmobilized in a fibrous bed bioreactor. AB - R-2-hydroxy-4-phenylbutyric acid (R-HPBA) is an important intermediate in the manufacture of angiotensin converting enzyme inhibitors. In this work, a recombinant D-lactate dehydrogenase (LDH) was used to transform 2-oxo-4 phenylbutyric acid (OPBA) to R-HPBA, with concomitant oxidation of beta nicotinamide adenine dinucleotide (NADH) to NAD(+). The cofactor NADH was regenerated by formate dehydrogenase (FDH) present in whole cells of Candida boidinii, which were pre-treated with toluene to make them permeable. The whole cells used in the process were more stable and easier to prepare as compared with the isolated FDH from the cells. Kinetic study showed that the reaction rate was dependent on the concentration of cofactor, NAD(+), and that both R-HPBA and OPBA inhibited the reaction. A novel method for co-immobilization of whole cells and LDH enzyme on cotton cloth was developed using polyethyleneimine (PEI), which induced the formation of PEI-enzyme-cell aggregates and their adsorption onto cotton cloth, leading to multilayer co-immobilization of cells and enzyme with high loading (0.5 g cell and 8 mg LDH per gram of cotton cloth) and activity yield ( > 95%). A fibrous bed bioreactor with co-immobilized cells and enzyme on the cotton cloth was then evaluated for R-HPBA production in fed-batch and repeated batch modes, which gave relatively stable reactor productivity of 9 g/L . h and product yield of 0.95 mol/mol OPBA when the concentrations of OPBA and R HPBA were less than 10 g/L. PMID- 16037988 TI - Enrichment using antibody-coated microfluidic chambers in shear flow: model mixtures of human lymphocytes. AB - Isolation of phenotypically-pure cell subpopulations from heterogeneous cell mixtures such as blood is a difficult yet fundamentally important task. Current techniques such as fluorescent activated cell sorting (FACS) and magnetic activated cell sorting (MACS) require pre-incubation with antibodies which lead to processing times of at least 15-60 min. In this study, we explored the use of antibody-coated microfluidic chambers to negative deplete undesired cell types, thus obtaining an enriched cell subpopulation at the outlet. We used human lymphocyte cell lines, MOLT-3 and Raji, as a model system to examine the dynamic cell binding behavior on antibody coated surfaces under shear flow. Shear stress ranging between 0.75 and 1.0 dyn/cm2 was found to provide most efficient separation. Cell adhesion was shown to follow pseudo-first order kinetics, and an anti-CD19 coated (Raji-depletion) device with approximately 2.6 min residence time was demonstrated to produce 100% pure MOLT-3 cells from 50-50 MOLT-3/Raji mixture. We have developed a mathematical model of the separation device based on the experimentally determined kinetic parameters that can be extended to design future separation modules for other cell mixtures. We conclude that we can design microfluidic devices that exploits the kinetics of dynamic cell adhesion to antibody coated surfaces to provide enriched cell subpopulations within minutes of total processing time. PMID- 16037989 TI - Genomic search for prostate cancer predisposition loci in Utah pedigrees. AB - BACKGROUND: We report a genome linkage scan in extended Utah pedigrees, utilizing a pedigree-splitting approach to reduce intra-familial heterogeneity. METHODS: Fifty-nine pedigrees with at least four Prostate cancer (PrCa) cases and no more than two meioses separating PrCa cases were analyzed using the CIDR genomic search STRP marker set. Parametric linkage analyses using dominant and recessive models were performed on four datasets resulting from a pedigree splitting algorithm. In addition, age at diagnosis subset analyses were performed. RESULTS: Four regions of interest (LODs>1.9) were identified on chromosomes 1p, 3q, 5q, and 22q. The linkage peaks on 1p, 3q, and 22q have been previously implicated for PrCa, though not significantly. The 1p region was supported by a single large Utah pedigree with a multipoint LOD score of 3.1. An additional 10 regions gave LOD scores>1.22 (nominal linkage evidence), including moderate evidence supporting the HPC20 region with a recessive model. CONCLUSIONS: Our genome-wide search in the informative, extended Utah pedigrees continues to illustrate an ability to identify and replicate linkage peaks, and supports four regions of interest for PrCa predisposition genes. PMID- 16037990 TI - Cement lines of secondary osteons in human bone are not mineral-deficient: new data in a historical perspective. AB - Using qualitative backscattered electron (BSE) imaging and quantitative energy dispersive X-ray (EDX) spectroscopy, some investigators have concluded that cement (reversal) lines located at the periphery of secondary osteons are poorly mineralized viscous interfaces with respect to surrounding bone. This conclusion contradicts historical observations of apparent highly mineralized (or collagen deficient) cement lines in microradiographs. Such conclusions, however, may stem from unrecognized artifacts that can occur during scanning electron microscopy. These include specimen degradation due to high-energy beams and the sampling of electron interaction volumes that extend beyond target locations during EDX analysis. This study used quantitative BSE imaging and EDX analysis, each with relatively lower-energy beams, to test the hypothesis that cement lines are poorly mineralized. Undemineralized adult human femoral diaphyses (n = 8) and radial diaphyses (n = 5) were sectioned transversely, embedded in polymethyl methacrylate, and imaged in a scanning electron microscope for BSE and EDX analyses. Unembedded samples were also evaluated. Additional thin embedded samples were stained and evaluated with light microscopy and correlated BSE imaging. BSE analyses showed the consistent presence of a bright line (higher atomic number) coincident with the classical location and description of the cement line. This may represent relative hypermineralization or, alternatively, collagen deficiency with respect to surrounding bone. EDX analyses of cement lines showed either higher Ca content or equivalent Ca content when compared to distant osteonal and interstitial bone. These data reject the hypothesis that cement lines of secondary osteons are poorly mineralized. PMID- 16037991 TI - Natural history of human prostate gland: Morphometric and histopathological analysis of Japanese men. AB - BACKGROUND: To clarify the pathology of the development of prostatic disorders such as inflammation, cancer, and hyperplasia, we compared histopathological findings of the prostate according to age group. METHODS: Whole-mount sections of prostates were used to assess the relationship between age and prostate weight (n=962), prostate histological composition in the transition zone (TZ) and in the peripheral zone (PZ) (n=68), prostate histopathological findings by zone (n=102), and comparison of latent tumor development by age group (n=1,815). RESULTS: A rapid increase in prostate weight from birth to the 20s was followed by a slow rise thereafter. Volume increases (P<0.01) were observed in all components of glandular epithelium, glandular lumen, and stroma in the TZ from the 40s to 70s inclusive. In the PZ, the epithelial and stromal volumes tended to decrease in an age-dependent manner (P<0.05). Calculi and lymphocyte infiltration were detected at a relatively early age, with a tendency towards an age-dependent increase. Glandular dilation and nodular hyperplasia were noted first in the 30s group, also with a tendency towards age-dependent increase. Latent tumors were first detected in the 30s group (5.6%), and slowly increased thereafter. CONCLUSIONS: There was an age-dependent trend towards prostate glandular dilation and prostate enlargement with inflammation. It was demonstrated that tumor and hyperplasia have a long natural history, usually starting in the fourth decade of life, accompanied by dynamic changes with age in glandular tissue composition as well as cell proliferation activity. PMID- 16037992 TI - Profiling of gene expression changes caused by p53 gain-of-function mutant alleles in prostate cancer cells. AB - BACKGROUND: Tumor suppressor p53 mutations are associated with the transition of prostate cancer to metastatic, hormone-refractory disease and stable expression of p53 gain-of-function (p53GOF) alleles support growth of LNCaP in androgen depleted medium. In this study, we performed gene expression profiling of four LNCaP-p53GOF sublines to test the hypothesis that different p53GOF mutants mediated androgen independence via modulation of a common set of genes. METHODS: Expression profiling was performed using Affymetrix HG-U95Av2 arrays followed by hierarchical clustering to identify expression patterns associated with particular molecular alterations. p53GOF-mediated regulation of Id-1 expression was validated by RT-PCR and dual-luciferase reporter assays. RNA interference was used to investigate the effects of Id-1 and Id-3 suppression. RESULTS: LNCaP p53GOF sublines possessed a molecular signature consisting of 95 differentially regulated genes that could be segregated into two clusters of transcripts induced (n=50) and repressed (n=45) by p53GOF expression. To begin validating these genes as effectors of the p53 mutants, we evaluated one of the overexpressed genes, Id 1. RT-PCR confirmed the microarray results and revealed elevated Id-1 levels in LNCaP-p53-P151S (loss-of-function only mutant), thereby implicating p53 mutational inactivation, but not gain-of-function, as a basis for Id-1 deregulation. Reporter assays demonstrated enhanced Id-1 promoter activity in an LNCaP-p53GOF subline. The contribution of Id-1 to p53GOF-mediated biology was demonstrated by the ability of RNAi-mediated gene silencing to decrease both basal and androgen-independent proliferation. CONCLUSIONS: While different p53GOF mutants result in overall distinct expression profiles, they share a common set of differentially-expressed genes that can be used to signify their presence and provide insight into mechanisms underlying androgen independence. PMID- 16037993 TI - Exploration of peptide motifs for potent non-viral gene delivery highly selective for dividing cells. AB - BACKGROUND: The immunogenicity of viral DNA vectors is an important problem for gene therapy. The use of peptide motifs for gene delivery would largely overcome this problem, and provide a simple, safe and powerful approach for non-viral gene therapy. METHODS: We explored the functional properties of two motifs: the (Lys)(16) motif (for binding and condensing DNA, and probably also nuclear translocation of plasmids) and the fusogenic peptide motif of influenza virus (for acid-dependent endocytic escape of peptide/DNA particles). The physical properties and gene delivery efficiencies of (Lys)(16)-containing peptides in combination with free fusogenic peptide were evaluated, and compared with a single composite peptide incorporating both moieties. Post-mitotic corneal endothelial cells and growth-arrested HeLa were included, so as not to neglect the question of nuclear translocation of plasmids. RESULTS: The fusogenic moiety in the composite peptide was able to adopt an alpha-helical configuration unhindered by the (Lys)(16) moiety, and retained acid-dependent fusogenic properties. The composite peptide gave remarkably high levels of gene delivery to dividing cell lines. However, in marked contrast to (Lys)(16)/DNA complexes plus free fusogenic peptide, the composite peptide was completely ineffective for gene delivery to post-mitotic and growth-arrested cells. CONCLUSIONS: Attachment of the fusogenic peptide to (Lys)(16) appears to block (Lys)(16)-mediated nuclear translocation of plasmid, but not fusogenic peptide mediated endocytic escape. This strengthens the experimental basis for (Lys)(16)-mediated nuclear translocation of plasmids, and provides a single peptide with potent gene delivery properties, restricted to dividing cells. This property is potentially useful in experimental biology and clinical medicine. PMID- 16037994 TI - Platinum-mediated oxidative addition and reductive elimination of imidazolium salts at C4 and C5. PMID- 16037995 TI - Molecular recognition of RNA by neomycin and a restricted neomycin derivative. PMID- 16037996 TI - Phenoxyl radicals hydrogen-bonded to imidazolium: analogues of tyrosyl D. of photosystem II: high-field EPR and DFT studies. PMID- 16037997 TI - A half adder based on a photochemically driven [2]rotaxane. PMID- 16037998 TI - Conservation of chirality in a hierarchical supramolecular self-assembled structure with pentagonal symmetry. PMID- 16037999 TI - pH-Switchable through-space interaction of organic radicals within a self assembled coordination cage. PMID- 16038000 TI - Stimuli-responsive controlled-release delivery system based on mesoporous silica nanorods capped with magnetic nanoparticles. PMID- 16038001 TI - Are hormones from the neuropeptide Y family recognized by their receptors from the membrane-bound state? AB - Hormones and many other neurotransmitters, growth factors, odorant molecules, and light all present stimuli for a class of membrane-anchored receptors called G protein-coupled receptors (GPCRs). The GPCRs are the largest family of cell surface receptors involved in signal transduction. About 1% of all known genes of Drosophila and more than 5% of the genes of Caenorhabditis elegans encode GPCRs. In addition, more than 50% of current therapeutic agents on the market target these receptors. When the enormous biological and pharmaceutical importance of these receptors is considered, it is surprising how little is known about the mechanism with which these receptors recognize their natural ligands. In this review we present a structural approach, utilizing techniques of high-resolution NMR spectroscopy, to address the question of whether peptides from the neuropeptide Y family of neurohormones are recognized directly from solution or from the membrane-bound state. In our studies we discovered that the structures of the membrane-bound species are better correlated to the pharmacological properties of these peptides than the solution structures are. These findings are supported by the observation that many biophysical properties of these peptides seem to be optimized for membrane binding. We finally present a scenario of possible events during receptor recognition. PMID- 16038002 TI - Probing the binding entropy of ligand-protein interactions by NMR. PMID- 16038003 TI - A gold catalyst for carbene-transfer reactions from ethyl diazoacetate. PMID- 16038004 TI - Preparation and characterization of inclusion complexes of beta-cyclodextrin with ionic liquid. AB - The solubilities of beta-cyclodextrin (beta-CD), ionic liquid (IL) 1-butyl-3 methylimidazolium hexafluorophosphate (bmimPF6), and their mixture in water were determined, and the conductivity of these aqueous solutions was measured. It was demonstrated that beta-CD and bmimPF6 could enhance the solubility of each other, and the solubility curves of each were linear with gradients of about 1. The conductivity decreased remarkably with increasing beta-CD concentration, and a discernible break in the conductivity curve could be observed when beta-CD and bmimPF6 were equimolar in the solution. The solubility and conductivity results indicated that inclusion complexes (ICs) of 1:1 stoichiometry were formed. The inclusion compounds were further characterized by using powder X-ray diffraction (XRD) analysis, 13C CP/MAS (cross-polarization magic-angle spinning) NMR and 1H NMR spectroscopy, and thermogravimetric analysis (TGA). The results showed that the ICs were a fine crystalline powder. The host-guest system exhibited a channel type structure and each glucose unit of beta-CD was in a similar environment. The decomposition temperature of the ICs was lower than that of bmimPF6 and beta-CD individually. PMID- 16038005 TI - Preparation, structure, and ethylene polymerization behavior of half-sandwich picolyl-functionalized carborane iridium, ruthenium, and rhodium complexes. AB - The synthesis of half-sandwich transition-metal complexes containing the Cab(N) and Cab(N,S) chelate ligands (HCab(N) = HC2B10H10CH2C5H4N (1), LiCab(N,S) = LiSC2B10H10CH2C5H4N (4)) is described. Compounds 1 and 4 were treated with chloride-bridged dimers [{Ir(Cp*)Cl2}2] (Cp* = eta5-C5Me5), [{Ru(p-cymene)Cl2}2] and [{Rh(Cp*)Cl2}2] to give half-sandwich complexes [Ir(Cp*)Cl(Cab(N))] (2), [Ru(p-cymene)Cl(Cab(N))] (3), and [Rh(Cp*)Cl(Cab(N,S))] (5), respectively. Addition reaction of LiCab(S) (Cab(S) = SC2(H)B10H10) to the rhodium complex 5 yields [Rh(Cp*)(Cab(S))(Cab(N,S))] (6). All the complexes were characterized by IR and NMR spectroscopy, and by elemental analysis. In addition, X-ray structure analyses were performed on complexes 2, 3, 5, and 6, in which the potential C,N- and N,S-chelate ligands were found to coordinate in a bidentate mode. The carborane complex 2 shows catalytic activities up to 3.7x10(5) g PE mol(-1) Ir h( 1) for the polymerization of ethylene in the presence of methylaluminoxane (MAO) as cocatalyst. The polymer obtained from this homogeneous catalytic reaction has a spherical morphology. Catalytic activities and the molecular weight of polyethylene have been investigated for various reaction conditions. PMID- 16038006 TI - PNA encoding (PNA=peptide nucleic acid): from solution-based libraries to organized microarrays. AB - Microarray-based technologies have attracted attention in chemical biology by virtue of their miniaturized format, which is well suited to probe ligand-protein interactions or investigate enzymatic activity in complex biological mixtures. A number of research groups have reported the preparation of surfaces on microarrays with specific functional groups to chemoselectively attach small molecules from libraries. We have developed an alternative method whereby libraries are encoded with peptide nucleic acid (PNA), such that libraries which exist as mixtures in solution self-assemble into an organized microarray through hybridization to produce readily available DNA arrays. This allows libraries synthesized by split and mix methods to be decoded in a single step. An asset of this method compared to direct spotting is that libraries can be used in solution for bioassays prior to self-assembly into the microarray format. PMID- 16038007 TI - 2-Bromophospholide ions: synthesis and theoretical study. AB - DFT calculations at the B3 LYP/6-311++G(3df,2p) level indicate that the 2 bromophospholide ion could be stable toward self-arylation as a result of the lowered nucleophilicity of the in-plane phosphorus lone pair (the corresponding sigma(P) orbital is lowered by 0.7 eV compared with the corresponding orbital of the parent phospholide ion, and the negative charge at P is reduced from -0.435 to -0.369 e). Accordingly, the synthesis of 2-bromo-3,4-dimethylphospholide was successfully carried out by quantitative base-induced dealkylation of 2-bromo-1 (2-ethoxycarbonylethyl)-3,4-dimethylphosphole. This ion reacts with FeCl2 to give the corresponding 2,2'-dibromo-3,3',4,4'-tetramethyl-1,1'-diphosphaferrocene as a poorly stable mixture of meso- and rac-diastereomers in 18% yield. PMID- 16038008 TI - Iodine monochloride-amine complexes: an experimental and computational approach to new chiral electrophiles. AB - Lactonizations are important steps in many synthetic sequences. Substrate controlled reactions that use chiral auxiliaries or chiral alkenes have already been studied in depth. This study focuses on stereoselective reagent-controlled iodolactonizations, by application of a new method that uses complexes of iodine monochloride and various donor molecules. (R)-1,2,3,4-Tetrahydro-1-naphthylamine and other amines with similar structures were found to be efficient in the iodocyclization of 4-aryl-4-pentenoic acids. Calculations were performed on complexes of (R)-1,2,3,4-tetrahydro-1-naphthylamine with XCl (X = I, H) to identify possible reactive species in these iodocyclizations. Calculations were carried out at various levels of theory, including B3 LYP/6-31+G (d,p) by using a modified SDD basis set for iodine. PMID- 16038009 TI - Trimethylyttrium and trimethyllutetium. PMID- 16038010 TI - Open-framework chalcogenides as visible-light photocatalysts for hydrogen generation from water. PMID- 16038011 TI - Ethnic differences in prenatal growth and the association with maternal and fetal characteristics. AB - OBJECTIVES: The objectives of this study were to determine ethnic differences in prenatal growth and to examine their association with differences in maternal and fetal characteristics such as maternal height, weight, age, parity and fetal gender. METHODS: A total of 1494 women from Rotterdam, The Netherlands, with a low-risk pregnancy who participated in a population-based cohort study, the Generation R Study, were offered three ultrasound examinations during pregnancy. Multilevel modeling was applied to determine ethnic differences in (estimated) fetal weight (including birth weight) and in the separate biometric variables that were used to calculate the estimated fetal weight (abdominal circumference, head circumference and femur length). Additionally the association of ethnic differences with maternal and fetal characteristics (i.e. maternal weight, height, age, parity and fetal gender) was studied. RESULTS: Turkish, Cape Verdian, Surinamese-Creole and Surinamese-Hindustani women had on average smaller fetuses than the native Dutch women. The differences became more pronounced towards term. In the Turkish group the differences were no longer statistically significant when adjusted for maternal weight, height, age, parity and fetal gender. In the Cape Verdian, Surinamese-Creole and Surinamese-Hindustani groups the differences decreased after adjustment (31%, 16% and 39%, respectively). CONCLUSIONS: This study shows that there are ethnic differences in fetal growth, which to a large extent may be attributed to differences in maternal weight, height, age and parity. For some ethnic groups, however, additional factors are involved, as differences remain significant after correction for fetal and maternal characteristics. PMID- 16038012 TI - A novel approach for gene therapy: engraftment of fibroblasts containing the artificial chromosome expression system at the site of inflammation. AB - BACKGROUND: Rheumatoid arthritis is characterized by inflammation of the synovial tissue. High systemic doses are necessary to achieve therapeutic levels of anti rheumatic drugs in the joints. Gene transfer might provide a more efficient delivery system for genes encoding therapeutic proteins. METHODS: The artificial chromosome expression system (ACE System) is a new non-integrating, non-viral gene expression system which functions like a natural chromosome. This technology offers advantages over current expression systems because it allows stable and predictable expression of proteins encoded by single or multiple genes over long periods of time. We are developing ex vivo gene therapy using murine artificial chromosomes containing a reporter gene (LacZ and red fluorescent protein (RFP)) for local delivery of genes in rats with adjuvant arthritis (AA). RESULTS: The delivery of the intact ACE System into rat fibroblast-like synoviocytes (FLS) and rat skin fibroblasts (RSF) was detected within 24 to 48 h post-transfection. After growing cells under selection, clones expressing LacZ and RFP were identified. Furthermore, we investigated the feasibility of local delivery of a reporter gene to the joints of rats with AA by ex vivo gene therapy. This resulted in engraftment of the injected cells in the synovial tissue microarchitecture and expression of the reporter gene. CONCLUSIONS: This work demonstrates the potential feasibility of treating arthritis and other inflammatory diseases using fibroblasts containing the ACE System as a non-viral vector for gene therapy. PMID- 16038013 TI - Neonatal outcome of antenatally diagnosed congenital cystic adenomatoid malformations. AB - OBJECTIVE: To investigate the natural history and outcome of antenatally diagnosed congenital cystic adenomatoid malformation (CCAM) of the lung. METHODS: This was a retrospective study of all cases of fetal CCAM of the lung diagnosed antenatally. All cases were referred to a tertiary center for further management. A computer search identified all referred cases, and the records of these patients were examined to determine the pregnancy outcome. RESULTS: In a 4-year period, 34 cases of fetal CCAM were referred for further management. At presentation, all the cases were noted to be unilateral CCAMs and the majority (79%) were microcystic in nature. The CCAMs were complicated by varying degrees of mediastinal shift (79%) and hydrops fetalis (18%). During the course of the pregnancy, the lung lesion was seen to reduce in size or resolve spontaneously in 76% of cases without any prenatal intervention (including resolution of hydrops in three cases). The overall survival rate into infancy was 88%. One pregnancy was terminated for persisting hydrops fetalis and another resulted in infant death from complications of neonatal cardiac surgery for an associated aortic coarctation. CONCLUSION: The outcome of antenatally detected CCAM is much better than previously reported even when complicated by hydrops fetalis at presentation. The latter seems to be related to the high spontaneous regression rate of this tumor. Despite the antenatal resolution of CCAMs on ultrasound, postnatal follow-up is recommended in view of the long-term complications of this malformation. PMID- 16038014 TI - Ethnic variation in the prevalence of echogenic intracardiac foci and the association with Down syndrome. AB - OBJECTIVE: To determine whether the prevalence of fetal echogenic intracardiac foci (EIF) differs according to maternal ethnicity. METHODS: We performed a retrospective cohort study of all women undergoing second-trimester diagnostic ultrasound examination and amniocentesis at a prenatal diagnosis referral center from January 1 2000 to July 1 2003. Data were collected on the presence of EIF, gestational age at time of ultrasound scan, karyotype results, maternal age and ethnicity. Univariate and multivariate analyses of EIF, ethnicity and presence of aneuploidy were conducted. RESULTS: Among the 7480 women qualifying for the study, EIF were found in 309 (4.1%). When maternal ethnicity was subdivided into Caucasian, African-American, Hispanic, Asian-American, Native American, Asian Indian, and Middle Eastern, the highest rates of EIF were found in fetuses of African-American (6.7%), Asian-American (6.9%), and Middle Eastern (8.1%) mothers compared to a rate of 3.3% in Caucasians (P < 0.001). In all ethnic groups except Hispanics, EIF was associated with an increased risk for Down syndrome (odds ratio range from 1.8 to 15.7). CONCLUSIONS: African-American, Asian-American, and Middle Eastern patients are more likely than patients of other ethnicities to have a fetus with an EIF. Even controlling for ethnicity, fetuses with an EIF still have an increased risk for Down syndrome. As more data accumulate, the prevalence of EIF and its association with Down syndrome among different ethnic groups can be incorporated into patient counseling. PMID- 16038015 TI - How to identify twins at low risk of spontaneous preterm delivery. AB - OBJECTIVE: The aim of this study was to evaluate transvaginal sonographic assessment of cervical length at 23 weeks as a screening test for spontaneous preterm delivery in order to define a cut-off value that could be used to select twin pregnancies at low risk of spontaneous preterm delivery. METHODS: In a prospective multicenter study of 383 twin pregnancies included before 14 + 6 weeks a cervical scan with measurement of the cervical length was performed at 23 weeks' gestation. The results were blinded for the clinicians if the cervical length was > or = 15 mm. The rates of spontaneous delivery at different cut-off levels of cervical length were determined. RESULTS: Eighty-nine percent of the twins had dichorionic placentation and 58% were conceived after assisted reproduction. The rate of spontaneous preterm delivery was 2.3% (1.5% for dichorionic (DC) and 9.1% for (MC) monochorionic twins) before 28 weeks and 18.5% (17.1% for DC and 29.5% for MC twins) before 35 weeks. The screen-positive rate was 5% for a cervical length < or = 20, 7-8% at < or = 25, 16-17% at < or = 30 and 34-48% at < or = 35 mm depending on chorionicity. The false-negative rate (1 negative predictive value) ranged from 1.2% at 28 weeks to 18.6% at 35 weeks for all twins. Receiver-operating characteristics curves showed that the sensitivity increased with declining gestational age with cut-off levels of highest accuracy at 21 mm for 28 weeks and 29 mm for 33 weeks. CONCLUSIONS: Cervical length measurement at 23 weeks of gestation is a good screening test for predicting twins at low risk of preterm and very preterm delivery, especially in DC twins. The present results suggest that a cut-off of 25 mm should be recommended. PMID- 16038016 TI - Atypical ductus venosus blood flow pattern in fetuses with severe tricuspid valve regurgitation. AB - We observed seven cases of atypical ductus venosus (DV) blood flow velocity waveform pattern with impairment of systolic forward flow resulting in a notch or a significant reduction in peak velocity during the S-wave in systole. All affected fetuses had severe tricuspid valve regurgitation associated with congestive heart failure and/or cardiac malformations. The decrease in venous systolic forward flow modulates the venous pulsatility indices towards more favorable values and should be considered when fetuses with tricuspid regurgitation are followed by Doppler assessment of the DV. Detection of these changes in the DV flow profile should prompt detailed color Doppler echocardiography with special emphasis on right atrioventricular valve regurgitation. PMID- 16038017 TI - Carbonylation of milk powder proteins as a consequence of processing conditions. AB - During industrial treatments, milk proteins could be oxidatively modified, thus leading to the formation of modified/oxidised amino acid residues. The apparition of such modified residues may contribute to the formation of new immunologically reactive structures. Some of these adducts could, in an advanced stage, lead to cross-linked protein species whose proteolytic susceptibility would be drastically decreased. Such protein species, that are resistant to digestion, could also constitute major food allergens. Therefore, these oxidative protein modifications tend to increase the natural allergenicity of milk proteins. For these reasons, monitoring milk protein oxidative modifications could be very useful regarding both product quality and allergenicity issues. In the present paper, we highlight, using different analytical approaches, the preferential carbonylation of beta-lactoglobulin (beta-Lg) during industrial treatments of milk. This result is particularly interesting since native beta-Lg represents one of the major milk allergens. PMID- 16038018 TI - Global detection and characterization of hypothetical proteins in Shewanella oneidensis MR-1 using LC-MS based proteomics. AB - The availability of whole genome sequences has enabled the application of powerful tools for assaying global expression patterns in environmentally relevant bacteria such as Shewanella oneidensis MR-1. A large number of genes in prokaryote genomes, including MR-1, have been annotated as hypothetical, indicating that no similar protein has yet been identified in other organisms. Using high-sensitivity MS coupled with accurate mass and time (AMT) tag methodology, 1078 tryptic peptides were collectively detected in MR-1 cultures, 671 of which were unique to their parent protein. Using only these unique tryptic peptides and a minimum of two peptides per protein, we identified, with high confidence, the expression of 258 hypothetical proteins. These proteins ranged from 3.5 to 139 kDa, with 47 being 100 amino acid residues or less. Using a combination of information including detection in cells grown under specific culture conditions, presence within a specific cell fraction, and predictive algorithms such as PSORT and PSORT-B, possible/plausible functions are proposed for some hypothetical proteins. Further, by applying this approach a number of proteins were found not only to be expressed, but only expressed under certain culturing conditions, thereby suggesting function while at the same time isolating several proteins to distinct locales of the cell. These results demonstrate the utility of the AMT tag methodology for comprehensive profiling of the microbial proteome while confirming the expression of a large number of hypothetical genes. PMID- 16038019 TI - The human platelet proteome mapped by peptide-centric proteomics: a functional protein profile. AB - Several studies have been published in which holistic approaches were used to characterise the proteome and transcriptome of human platelets. The key intent being that a deeper understanding of the normal and aberrant physiological functions of platelets can only be achieved if most biomolecular building blocks are mapped. Here we present the application of recently developed novel technologies that overcome some of the shortcomings of gel-based proteomics. Central in our approach is the so-called combined fractional diagonal chromatography (COFRADIC)-technology in which sets of representative peptides are sorted in a diagonal RP chromatographic system through a specific modification of their side chain. In this study we combined three different COFRADIC sorting techniques to analyse the proteome of human platelets. Methionyl, cysteinyl and amino terminal peptides were isolated and analysed by MS/MS. Merging the peptide identifications obtained after database searching resulted in a core set of 641 platelet proteins, which comprises the largest set identified today. In comparison to previously published platelet proteomes, we identified 404 novel platelet proteins containing a high number of hydrophobic membrane proteins and hypothetical proteins. Furthermore we discuss the observed characteristics and potential benefits of each of the different COFRADIC technologies for proteome analysis and highlight important issues that need to be considered when searching sequence databases using data obtained in peptide-centric, non-gel proteomics studies. PMID- 16038020 TI - Proteome profiling of interleukin-12 treated human T helper cells. AB - Selective activation of T helper subsets 1 (Th1) and 2 (Th2) plays a crucial role in different pathological conditions. Th1 cell response is involved in pathogenesis of autoimmune diseases, such as type II diabetes and multiple sclerosis, and Th2 cell response in pathogenesis of allergy and asthma. Cytokine interleukin-12 (IL-12) is one of the key factors in the differentiation of naive CD4(+) T cells into Th1 cells. In this study we used 2-DE and MS to find and identify IL-12 regulated proteins in human CD4(+) T cells. In total, 42 protein spots were found to be differentially expressed following IL-12 stimulation, of which 22 were up- and 20 down-regulated. Among the upregulated proteins there are a multifunctional cytokine macrophage migration inhibitory factor and a known IL 12 target gene Programmed cell death 4. Downregulated proteins include p21 activated kinase 2 and its upstream GTPase Cdc42. Compared to previous reports our analysis provides a new view on the IL-12 induced changes on CD4(+) T cells underscoring the importance of creating and combining the data generated at various levels to build a comprehensive view of a given biological process of the cell. PMID- 16038021 TI - Peptidomic analysis of human blood specimens: comparison between plasma specimens and serum by differential peptide display. AB - The human Plasma Proteome Project pilot phase aims to analyze serum and plasma specimens to elucidate specimen characteristics by various proteomic techniques to ensure sufficient sample quality for the HUPO main phase. We used our proprietary peptidomics technologies to analyze the samples distributed by HUPO. Peptidomics summarizes technologies for visualization, quantitation, and identification of the low-molecular-weight proteome (<15 kDa), the "peptidome." We analyzed all four HUPO specimens (EDTA plasma, citrate plasma, heparin plasma, and serum) from African- and Asian-American donors and compared them to in-house collected Caucasian specimens. One main finding focuses on the most suitable method of plasma specimen collection. Gentle platelet removal from plasma samples is beneficial for improved specificity. Platelet contamination or activation of platelets by low temperature prior to their removal leads to distinct and multiple peptide signals in plasma samples. Two different specimen collection protocols for platelet-poor plasma are recommended. Further emphasis is placed on the differences between plasma and serum on a peptidomic level. A large number of peptides, many of them in rather high abundance, are only present in serum and not detectable in plasma. This ex vivo generation of multiple peptides hampers discovery efforts and is caused by a variety of factors: the release of platelet derived peptides, other peptides derived from cellular components or the clot, enzymatic activities of coagulation cascades, and other proteases. We conclude that specimen collection is a crucial step for successful peptide biomarker discovery in human blood samples. For analysis of the low-molecular-weight proteome, we recommend the use of platelet-depleted EDTA or citrate plasma. PMID- 16038022 TI - Immunoassay and antibody microarray analysis of the HUPO Plasma Proteome Project reference specimens: systematic variation between sample types and calibration of mass spectrometry data. AB - Four different immunoassay and antibody microarray methods performed at four different sites were used to measure the levels of a broad range of proteins (N = 323 assays; 39, 88, 168, and 28 assays at the respective sites; 237 unique analytes) in the human serum and plasma reference specimens distributed by the Plasma Proteome Project (PPP) of the HUPO. The methods provided a means to (1) assess the level of systematic variation in protein abundances associated with blood preparation methods (serum, citrate-anticoagulated-plasma, EDTA anticoagulated-plasma, or heparin-anticoagulated-plasma) and (2) evaluate the dependence on concentration of MS-based protein identifications from data sets using the HUPO specimens. Some proteins, particularly cytokines, had highly variable concentrations between the different sample preparations, suggesting specific effects of certain anticoagulants on the stability or availability of these proteins. The linkage of antibody-based measurements from 66 different analytes with the combined MS/MS data from 18 different laboratories showed that protein detection and the quality of MS data increased with analyte concentration. The conclusions from these initial analyses are that the optimal blood preparation method is variable between analytes and that the discovery of blood proteins by MS can be extended to concentrations below the ng/mL range under certain circumstances. Continued developments in antibody-based methods will further advance the scientific goals of the PPP. PMID- 16038023 TI - Treatment of uncomplicated reflux disease. AB - Uncomplicated reflux disease comprises the non-erosive reflux disease (NERD) and erosive reflux disease (ERD). The objectives of treatment are the adequate control of symptoms with restoration of quality of life, healing of lesions and prevention of relapse. Treatment of NERD consists in the administration of proton pump inhibitors (PPI) for 2-4 wk, although patients with NERD show an overall poorer response to PPI treatment than patients with ERD owing to the fact that patients with NERD do not form a pathophysiologically homogenous group. For long term management on-demand treatment with a PPI is probably the best option. In patients with ERD, therapy with a standard dose PPI for 4-8 wk is always recommended. Long-term treatment of ERD is applied either intermittently or as continuous maintenance treatment with an attempt to reduce the daily dosage of the PPI (step-down principle). In selected patients requiring long-term PPI treatment, antireflux surgery is an alternative option. In patients with troublesome reflux symptoms and without alarming features empirical PPI therapy is another option for initial management. Therapy should be withdrawn after initial success. In the case of relapse, the long-term care depends on a careful risk assessment and the response to PPI therapy. PMID- 16038024 TI - Oral Xeloda plus bi-platinu two-way combined chemotherapy in treatment of advanced gastrointestinal malignancies. AB - AIM: To compare the effect, adverse events, cost-effectiveness and dose intensity (DI) of oral Xeloda vs calcium folinate (CF)/5-FU combination chemotherapy in patients with advanced gastrointestinal malignancies, both combined with bi platinu two-way chemotherapy. METHODS: A total of 131 patients were enrolled and randomly selected to receive either oral Xeloda (X group) or CF/5-FU (control group). Oral Xeloda 1,000 mg/m2 was administered twice daily from d 1 to 14 in X group, while CF 200 mg/m2 was taken as a 2-h intravenous infusion followed by 5 FU 600 mg/m2 intravenously for 4-6 h on d 1-5 in control group. Cisplatin and oxaliplatin were administered in the same way to both the groups: cisplatin 60-80 mg/m2 by hyperthermic intraperitoneal administration, and oxaliplatin 130 mg/m2 intravenously for 2 h on d 1. All the drugs were recycled every 21 d, with at least two cycles. Pyridoxine 50 mg was given t.i.d. orally for prophylaxis of the hand-foot syndrome (HFS). Then the effect, adverse events, cost-effectiveness and DI of the two groups were evaluated. RESULTS: Hundred and fourteen cases (87.0%) finished more than two chemotherapy cycles. The overall response rate of them was 52.5% (X group) and 42.4% (control group) respectively. Tumor progression time (TTP) was 7.35 mo vs 5.95 mo, and 1-year survival rate was 53.1% vs 44.5%. There was a remarkable statistical significance of TTP and 1-year survival between the two groups. The main Xeloda-related adverse events were myelosuppression, gastrointestinal toxicity, neurotoxicity and HFS, which were mild and well tolerable. Therefore, no patients withdrew from the study due to side effects before two chemotherapy cycles were finished. Both groups finished pre-arranged DI and the relative DI was nearly 1.0. The average cost for 1 patient in one cycle was RMB9 137.35 (X group) and RMB8 961.72 (control group), or USD1 100.89 in X group and USD1 079.73 in control group. To add 1% to the response rate costs RMB161.44 vs RMB210.37 respectively (USD19.45 vs USD25.35). One-month prolongation of TTP costs RMB1 243.18 vs RMB1 506.17 (USD149.78 vs USD181.47). Escalation of 1% of 1-year survival costs RMB172.74 vs RMB201.64 (USD20.75 vs USD24.29). CONCLUSION: Oral Xeloda combined with bi-platinu two-way combination chemotherapy is efficient and tolerable for patients with advanced gastrointestinal malignancies; meanwhile the expenditure is similar to that of CF/5-FU combined with bi-platinu chemotherapy, and will be cheaper if we are concerned about the increase of the response rate, TTP or 1-year-survival rate pharmacoeconomically. PMID- 16038025 TI - Effect of superoxide dismutase and malondialdehyde metabolic changes on carcinogenesis of gastric carcinoma. AB - AIM: To investigate the relationship between the superoxide dismutase (SOD), malondialdehyde (MDA) metabolic changes and the gastric carcinogenesis. METHODS: The SOD activity and MDA content were measured in the gastric tissues from the focus center, peripheral and far-end areas of gastric carcinoma (n = 52) and gastric ulcer (n = 10). All the tissues were subjected to routine histological examinations and classifications. RESULTS: The SOD activity was greatly reduced but the MDA content was markedly increased in the center areas of the non-mucous gastric carcinoma (non-MGC); and the poorly differentiated gastric carcinoma varied. The SOD activity was gradually decreased and the MDA content was gradually increased in the tissues from the focus far-end, peripheral to center areas of non-MGC. Both of the SOD activity and the MDA content were significantly declined and were respectively at same low level in the tissues from the focus center, peripheral, and far-end area with the mucous gastric carcinoma (MGC). In contrast to the gastric ulcer and grade I or II of non-MGC, the same level of the SOD activity and the MDA content were found in the focus center areas. Between non-MGC (groups A-D) and gastric ulcer (group F), the differences of SOD activity and MDA content were very noticeable in the gastric tissues from the focus peripheral and far-end areas, in which the SOD activity showed noticeable increase and the MDA content showed noticeable decrease in the gastric ulcer. CONCLUSION: The active free radical reaction in the gastric tissues can induce the carcinogenesis of non-MGC. The utmost low ability of antioxidation in the gastric tissues can induce the carcinogenesis of MGC. The metabolic change of the free radicals centralized mostly in the center of ulcerated lesions only, which suggested the ability of antioxidation was declined only in these lesions. However, the metabolism of free radicals varied significantly and the ability of antioxidation declined not only in the local focus area but also in the abroad gastric tissues with gastric carcinoma. PMID- 16038026 TI - Nutritional factors and gastric cancer in Zhoushan Islands, China. AB - AIM: To investigate the association between nutrient intakes and high incidence rate of gastric cancer among residents in Zhoushan Islands. METHODS: A frequency matched design of case-control study was used during the survey on dietary factors and gastric cancer in Zhoushan Islands, China. A total of 103 cases of gastric cancer diagnosed in 2001 were included in the study and 133 controls were randomly selected from the residents in Zhoushan Islands. A food frequency questionnaire was specifically designed for the Chinese dietary pattern to collect information on dietary intake. A computerized database of the dietary and other relative information of each participant was completed. Total calories and 15 nutrients were calculated according to the food composition table and their adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were estimated by gender using unconditional logistic regression models. RESULTS: High intakes of protein, saturated fat, and cholesterol were observed with the increased risk of gastric cancer particularly among males (OR(Q4 vs Q1) were 10.3, 3.24, 2.76 respectively). While carbohydrate was a significant high-risk nutrient (OR(Q4 vs Q1) = 14.8; P for linear trend = 0.024) among females. Regardless of their gender, the cases reported significantly higher daily intake of sodium mainly from salts. As to the nutrients of vitamins A and C, an inversed association with the risk of GC was found. Baseline characteristics of participants were briefly described. CONCLUSION: The findings from this study confirm the role of diet related exposure in the etiology of gastric cancer from the point of view of epidemiology. An increased risk of gastric cancer is associated with high intakes of protein, saturated fat, cholesterol and sodium, while consumption of polyunsaturated fat, vitamin A and ascorbic acid may have a protective effect against gastric cancer. PMID- 16038027 TI - Growth inhibition of high-intensity focused ultrasound on hepatic cancer in vivo. AB - AIM: To investigate the damaging effect of high-intensity focused ultrasound (HIFU) on cancer cells and the inhibitory effect on tumor growth. METHODS: Murine H22 hepatic cancer cells were treated with HIFU at the same intensity for different lengths of time and at different intensities for the same length of time in vitro, the dead cancer cells were determined by trypan blue staining. Two groups of cancer cells treated with HIFU at the lowest and highest intensity were inoculated into mice. Tumor masses were removed and weighed after 2 wk, tumor growth in each group was confirmed pathologically. RESULTS: The death rate of cancer cells treated with HIFU at 1 000 W/cm2 for 0.5, 1, 2, 4, 8, and 12 s was 3.11+/-1.21%, 13.37+/-2.56%, 38.84+/-3.68%, 47.22+/-5.76%, 87.55+/-7.32%, and 94.33+/-8.11%, respectively. A positive relationship between the death rates of cancer cells and the length of HIFU treatment time was found (r = 0.96, P<0.01). The death rate of cancer cells treated with HIFU at the intensity of 100, 200, 400, 600, 800, and 1 000 W/cm2 for 8 s was 26.31+/-3.26%, 31.00+/-3.87%, 41.97+/ 5.86%, 72.23+/-8.12%, 94.90+/-8.67%, and 99.30+/-9.18%, respectively. A positive relationship between the death rates of cancer cells and the intensities of HIFU treatment was confirmed (r = 0.98, P<0.01). The cancer cells treated with HIFU at 1 000 W/cm2 for 8 s were inoculated into mice ex vivo. The tumor inhibitory rate was 90.35% compared to the control (P<0.01). In the experimental group inoculated with the cancer cells treated with HIFU at 1 000 W/cm2 for 0.5 s, the tumor inhibitory rate was 22.9% (P<0.01). By pathological examination, tumor growth was confirmed in 8 out of 14 mice (57.14%, 8/14) inoculated with the cancer cells treated with HIFU at 1 000 W/cm2 for 8 s, which was significantly lower than that in the control (100%, 15/15, P<0.05). CONCLUSION: HIFU is effective on killing or damage of H22 hepatic cancer cells in vitro and on inhibiting tumor growth in mice ex vivo. PMID- 16038028 TI - Classification of right hepatectomy for special localized malignant tumor in right liver lobe. AB - AIM: To describe a new classification method of right hepatectomy according to the different special positions of tumors. METHODS: According to positions, 91 patients with malignant hepatic tumor in the right liver lobe were divided into six groups: tumors in the right posterior lobe and (or) the right caudate lobe compressing the right portal hilum (n = 14, 15.4%), tumors in the right liver lobe compressing the inferior vena cava and (or) hepatic veins (n = 11, 12.9%), tumors infiltrating diaphragmatic muscle (n = 7, 7.7%), tumors in the hepatorenal recess (infiltrating the right fatty renal capsule, transverse colon and right adrenal gland, n = 8, 8.8%), tumors deeply located near the vertebral body (n = 3, 3.3%), tumors at other sites in the right liver lobe (the control group, n = 48, 52.75%). The values of intraoperative blood loss (IBL), tumor's maxim cross section area (TMCSA), and time of hepatic hilum clamping (THHC) and incidence of postoperative complications were compared between five groups of tumor and control group, respectively. RESULTS: The THHC in groups 1-4 was significantly longer than that in the control group, the IBL in groups 1-4 was significantly higher than that in the control group, the TMCSA in groups 2-4 was significantly larger than that in the control group, and the ratio of IBL/TMCSA in group 1 was significantly higher than that in the control group. There was no significant difference in the indexes between group 5 and the control group. CONCLUSION: The site of tumor is the key factor that determines IBL. PMID- 16038029 TI - Apoptosis and its pathway in X gene-transfected HepG2 cells. AB - AIM: To investigate the effect of hepatitis B virus (HBV) X gene on apoptosis and expressions of apoptosis factors in X gene-transfected HepG2 cells. METHODS: The HBV X gene eukaryon expression vector pcDNA3-X was transiently transfected into HepG2 cells by lipid-media transfection. Untransfected HepG2 and HepG2 transfected with pcDNA3 were used as controls. Expression of HBx in HepG2 was identified by RT-PCR. MTT and TUNEL were employed to measure proliferation and apoptosis of cells in three groups. Semi-quantified RT-PCR was used to evaluate the expression levels of Fas/FasL, Bax/Bcl-xL, and c-myc in each group. RESULTS: HBV X gene was transfected into HepG2 cells successfully. RT-PCR showed that HBx was only expressed in HepG2/pcDNA3-X cells, but not expressed in HepG2 and HepG2/pcDNA3 cells. Analyzed by MTT, cell proliferation capacity was obviously lower in HepG2/pcDNA3-X cells (0.08910+/-0.003164) than in HepG2 (0.14410+/ 0.004927) and HepG2/pcDNA3 cells (0.12150+/-0.007159) (P<0.05 and P<0.01). Analyzed by TUNEL, cell apoptosis was much more in HepG2/pcDNA3-X cells (980/2,000) than HepG2 (420/2,000), HepG2/pcDNA3 cells (520/2,000) (P<0.05 and P<0.01). Evaluated by semi-quantified RT-PCR, the expression level of Fas/FasL was significantly higher in HepG2 cells transfected with HBx than in HepG2 and HepG2/pcDNA3 cells (P<0.05 and P<0.01). Bax/Bcl-xL expression level was also elevated in HepG2/pcDNA3-X cells (P<0.05 and P<0.01). Expression of c-myc was markedly higher in HepG2/pcDNA3-X cells than in HepG2 and HepG2/pcDNA3 cells (P<0.05 and P<0.01). CONCLUSION: HBV X gene can impair cell proliferation capacity, improve cell apoptosis, and upregulate expression of apoptosis factors. The intervention of HBV X gene on the expression of apoptosis factors may be a possible mechanism responsible for the change in cell apoptosis and proliferation. PMID- 16038030 TI - Treatment of metastatic colorectal carcinomas by systemic inhibition of vascular endothelial growth factor signaling in mice. AB - AIM: Tumor angiogenesis has been shown to be promoted by vascular endothelial growth factor (VEGF) via stimulating endothelial cell proliferation, migration, and survival. Blockade of VEGF signaling by different means has been demonstrated to result in reduced tumor growth and suppression of tumor angiogenesis in distinct tumor entities. Here, we tested a recombinant adenovirus, AdsFlt1-3, that encodes an antagonistically acting fragment of the VEGF receptor 1 (Flt-1), for systemic antitumor effects in pre-established subcutaneous CRC tumors in mice. METHODS: Murine colorectal carcinoma cells (CT26) were inoculated subcutaneously into Balb/c mice for in vivo studies. Tumor size and survival were determined. 293 cell line was used for propagation of the adenoviral vectors. Human lung cancer line A549 and human umbilical vein endothelial cells were transfected for in vitro experiments. RESULTS: Infection of tumor cells with AdsFlt1-3 resulted in protein secretion into cell supernatant, demonstrating correct vector function. As expected, the secreted sFlt1-3 protein had no direct effect on CT26 tumor cell proliferation in vitro, but endothelial cell function was inhibited by about 46% as compared to the AdLacZ control in a tube formation assay. When AdsFlt1-3 (5 x 10(9) PFU/animal) was applied to tumor bearing mice, we found a tumor inhibition by 72% at d 12 after treatment initiation. In spite of these antitumoral effects, the survival time was not improved. According to reduced intratumoral microvessel density in AdsFlt1-3-treated mice, the antitumor mechanism can be attributed to angiostatic vector effects. We did not detect increased systemic VEGF levels after AdsFlt1-3 treatment and liver toxicity was low as judged by serum alanine aminotransferase determination. CONCLUSION: In this study we confirmed the value of a systemic administration of AdsFlt1-3 to block VEGF signaling as antitumor therapy in an experimental metastatic colorectal carcinoma model in mice. PMID- 16038031 TI - Effect of NS398 on metastasis-associated gene expression in a human colon cancer cell line. AB - AIM: To investigate the effect of NS398 on the metastasis-associated gene expression in LoVo colorectal cancer cells. METHODS: LoVo cells were treated with NS398 at the concentration of 100 micromol/L for 24 and 48 h respectively. Total RNA was extracted with TRIzol reagents and reverse transcribed with Superscript II and hybridized with cDNA microarray (containing oncogenes, tumor suppressor genes, signal transduction molecules, adhesive molecules, growth factors, and ESTs) fabricated in our laboratory. After normalization, the ratio of gene expression of NS398 treated to untreated LoVo cells was either 2-fold up or 0.5 fold down was defined as the differentially expressed genes. Semi-quantitative RT PCR was used to validate the microarray results. RESULTS: Among the 447 metastasis-associated genes, 9 genes were upregulated and 8 genes were downregulated in LoVo cells treated with NS398 for 24 h compared to untreated cells. While 31 genes were upregulated and 14 genes were downregulated in LoVo cells treated with NS398 for 48 h. IGFBP-5, PAI-2, JUN, REL, BRCA1, and BRCA2 might be the new targets of NS398 in treatment of colorectal cancer. CONCLUSION: NS398 might exert its anti-metastasis effect on colorectal cancer by affecting several metastasis-associated gene expression. PMID- 16038032 TI - Outcome of lamivudine-resistant hepatitis B virus is generally benign except in cirrhotics. AB - AIM: We set to determine factors that determine clinical severity after the development of resistance. METHODS: Thirty-five Asian patients with genotypic lamivudine resistance were analyzed in three groups: 13/35 (37%) were non cirrhotics with normal pre-treatment ALT (Group IA), 12/35 (34%) were non cirrhotics with elevated pre-treatment ALT (Group IB), and 10/35 (29%) were cirrhotics (Group II). Patients were followed for a median of 98 wk (range 26 220) after the emergence of genotypic resistance. RESULTS: Group IA patients tended to retain normal ALT. Group IB patients showed initial improvement of ALT with lamivudine but 9/12 patients (75%) developed abnormal ALT subsequently. On follow-up however, this persisted in only 33%. Group II patients also showed improvement while on treatment, but they deteriorated with the emergence of resistance with 30% death from decompensated liver disease. Pretreatment ALT levels and CPT score (in the cirrhotic group) were predictive of clinical resistance and correlated with peak ALT levels and CPT score. CONCLUSION: The phenotype of lamivudine-resistant HBV correlated with the pretreatment phenotype. The clinical course was generally benign in non-cirrhotics. However, cirrhotics had a high risk of progression and death (30%) with the development of lamivudine resistance. PMID- 16038033 TI - Possible mechanism for hepatitis B virus X gene to induce apoptosis of hepatocytes. AB - AIM: To investigate the possible mechanism for HBV X gene to induce apoptosis of hepatocyte HL-7702 cells. METHODS: HBV X gene eukaryon expression vector pcDNA3-X was established and transfected into HL-7702 cells by lipid-mediated transfection, including transient and stable transfection. Positive clones were screened by incubating in the selective medium with 600 microg/mL G418 and named HL-7702/HBV-encoded X protein (HBx) cells. The expressions of Fas/FasL, Bax/Bcl 2, and c-myc mRNA were measured by semi-quantitative RT-PCR in HL-7702/HBx and control group, respectively. RESULTS: RT-PCR analysis confirmed that HBV X gene was transfected into HL-7702 cells successfully. By semi-quantitative RT-PCR analysis, Bax and c-myc mRNA levels in HL-7702/HBx cells of transient transfection were significantly higher than those in control, FasL and c-myc mRNA levels in HL-7702/HBx cells of stable transfection were significantly higher than those in control, whereas the Bcl-2 mRNA levels in HL-7702/HBx cells of transient and stable transfection were significantly lower than those in control. CONCLUSION: HBV X gene may promote the apoptosis of hepatocytes by regulating the expressions of Fas/FasL, Bax/Bcl-2, and c-myc gene in a dose-dependent manner. PMID- 16038034 TI - Expression of CD86 and increased infiltration of NK cells are associated with Helicobacter pylori-dependent state of early stage high-grade gastric MALT lymphoma. AB - AIM: A high percentage of early-stage high-grade gastric mucosa-associated lymphoid tissue (MALT) lymphomas remain Helicobacter pylori (H pylori)-dependent. However, unlike their low-grade counterparts, high-grade gastric MALT lymphomas may progress rapidly if unresponsive to H pylori eradication. It is mandatory to identify markers that may predict the H pylori-dependent status of these tumors. Proliferation of MALT lymphoma cells depends on cognate help and cell-to-cell contact of H pylori-specific intratumoral T-cells. To examine whether the expression of co-stimulatory marker CD86 (B7.2) and the infiltration of CD56 (+) natural killer (NK) cells can be useful markers to predict H pylori-dependent status of high-grade gastric MALT lymphoma. METHODS: Lymphoma biopsies from 26 patients who had participated in a prospective study of H pylori-eradication for stage I(E) high-grade gastric MALT lymphomas were evaluated. Tumors that resolved to Wotherspoon grade II or less after H pylori eradication were classified as H pylori-dependent; others were classified as H pylori-independent. The infiltration of NK cells and the expression of CD86 in pre-treatment paraffin embedded lymphoma tissues were determined by immunohistochemistry. RESULTS: There were 16 H pylori-dependent and 10 H pylori-independent cases. CD86 expression was detected in 11 (68.8%) of 16 H pylori-dependent cases but in none of 10 H pylori independent cases (P = 0.001). H pylori-dependent high-grade gastric MALT lymphomas contained significantly higher numbers of CD56 (+) NK cells than H pylori-independent cases (2.8+/-1.4% vs 1.1+/-0.8%; P = 0.003). CD86 positive MALT lymphomas also showed significantly increased infiltration of CD56 (+) NK cells compared to CD86-negative cases (2.9+/-1.1% vs 1.4+/-1.3%; P = 0.005). CONCLUSION: These results suggest that the expression of co-stimulatory marker CD86 and the increased infiltration of NK cells are associated with H pylori dependent state of early-stage high-grade gastric MALT lymphomas. PMID- 16038035 TI - Effect of Helicobacter pylori infection on p53 expression of gastric mucosa and adenocarcinoma with microsatellite instability. AB - AIM: To investigate the relationship between Helicobacter pylori (H pylori) infection, microsatellite instability and the expressions of the p53 in gastritis, intestinal metaplasia and gastric adenocarcinoma and to elucidate the mechanism of gastric carcinogenesis relating to H pylori infection. METHODS: One hundred and eight endoscopic biopsies and gastric adenocarcinoma were available for the study including 33 cases of normal, 45 cases of gastritis, 30 cases of intestinal metaplasia, and 46 cases of gastric adenocarcinoma. Peripheral blood samples of these patients were also collected. H pylori infection and p53 expressions were detected by means of streptavidin-peroxidase (SP) immunohistochemical method. Microsatellite loci were studied by PCR-SSCP-CE using the markers BAT-26, D17S261, D3S1283, D2S123, and D3S1611. MSI was defined as the peak shift in the DNA of the gastric tissue compared with that of the peripheral blood samples. Based on the number of mutated MSI markers, specimens were characterized as high MSI (MSI-H) if they manifested instability at two or more markers, low MSI (MSI-L) if unstable at only one marker, and microsatellite stable (MSS) if they showed no instability at any marker. RESULTS: H pylori infection was detected in the samples of gastritis, intestinal metaplasia, and gastric adenocarcinoma and the infection frequencies were 84.4%, 76.7%, and 65.2%, respectively, whereas no H pylori infection was detected in the samples of normal control. There was a significant difference in the infection rates between gastritis and carcinoma samples (P = 0.035). No MSI was detected in gastritis samples, one MSI-H and two MSI-L were detected among the 30 intestinal metaplasia samples, and 12 MSI-H and 3 MSI-L were detected in the 46 gastric carcinomas. In those gastric carcinomas, the MSI-H frequency in H pylori-positive group was significantly higher than that in H pylori-negative group. No p53 expression was detected in the normal and gastritis samples from dyspeptic patients. P53 positive immunohistochemical staining was detected in 13.3% of intestinal metaplasia samples and in 43.5% of gastric carcinoma samples. The levels of p53 in H pylori-positive samples were higher than those in the negative group when the carcinoma samples were subdivided into H pylori-positive and -negative groups (P = 0.013). Eight samples were detected with positive p53 expression out of the 11 MSI-H carcinomas with H pylori infection and no p53 expression could be seen in the H pylori-negative samples. CONCLUSION: H pylori affect the p53 pattern in gastric mucosa when MMR system fails to work. Mutations of the p53 gene seem to be an early event in gastric carcinogenesis. PMID- 16038036 TI - Sensory-motor responses to mechanical stimulation of the esophagus after sensitization with acid. AB - AIM: Sensitization most likely plays an important role in chronic pain disorders, and such sensitization can be mimicked by experimental acid perfusion of the esophagus. The current study systematically investigated the sensory and motor responses of the esophagus to controlled mechanical stimuli before and after sensitization. METHODS: Thirty healthy subjects were included. Distension of the distal esophagus with a balloon was performed before and after perfusion with 0.1 mol/L hydrochloric acid for 30 min. An impedance planimetry system was used to measure cross-sectional area, volume, pressure, and tension during the distensions. A new model allowed evaluation of the phasic contractions by the tension during contractions as a function of the initial muscle length before the contraction (comparable to the Frank-Starling law for the heart). Length-tension diagrams were used to evaluate the muscle tone before and after relaxation of the smooth muscle with butylscopolamine. RESULTS: The sensitization resulted in allodynia and hyperalgesia to the distension volumes, and the degree of sensitization was related to the infused volume of acid. Furthermore, a nearly 50% increase in the evoked referred pain was seen after sensitization. The mechanical analysis demonstrated hyper-reactivity of the esophagus following acid perfusion, with an increased number and force of the phasic contractions, but the muscle tone did not change. CONCLUSION: Acid perfusion of the esophagus sensitizes the sensory pathways and facilitates secondary contractions. The new model can be used to study abnormal sensory-motor mechanisms in visceral organs. PMID- 16038037 TI - Relationship between gastrointestinal and extra-gastrointestinal symptoms and delayed gastric emptying in functional dyspeptic patients. AB - AIM: Delayed gastric emptying and an enlarged fasting gastric antrum are common findings in functional dyspepsia but their relationship with gastrointestinal (GI), and the frequently associated extra-GI symptoms remains unclear. This study evaluated the relationship between GI and extra-GI symptoms, fasting antral volume and delayed gastric emptying in functional dyspepsia. METHODS: In 108 functional dyspeptic patients antral volume and gastric emptying were assessed with ultrasonography (US). Symptoms were assessed with standardized questionnaire. The association of symptoms and fasting antral volume with delayed gastric emptying was estimated with logistic regression analysis. RESULTS: Delayed gastric emptying was detected in 39.8% of the patients. Postprandial drowsiness (AOR 11.25; 95%CI 2.75-45.93), nausea (AOR 3.51; 95%CI 1.19-10.32), fasting antral volume (AOR 1.93; 95%CI 1.22-3.05), were significantly associated with delayed gastric emptying. Symptoms, mainly the extra-GI ones as postprandial drowsiness and nausea, combined with fasting antral volume predicted the modality of gastric emptying with a sensitivity and specificity of 78%. CONCLUSION: In functional dyspeptic patients, (1) an analysis of fasting antral volume and of symptoms can offer valuable indication on the modality of gastric emptying, and (2) it seems appropriate to inquire on postprandial drowsiness that showed the best correlation with delayed gastric emptying. PMID- 16038038 TI - Expression of adhesion molecules on mature cholangiocytes in canal of Hering and bile ductules in wedge biopsy samples of primary biliary cirrhosis. AB - AIM: To examine the expression of intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1) expression on canals of Hering (CoH) and bile ductules associated with the autoimmune process of bile duct destruction in primary biliary cirrhosis (PBC). METHODS: Ten wedged liver biopsies of PBC (five cases each of stages 2 and 3) were studied. The liver specimens were processed for transmission electron microscopy. Immunohistochemistry was performed using anti-ICAM-1 and anti-LFA-1 mouse mAbs. In situ hybridization was done to examine the messenger RNA expression of ICAM-1 in formalin-fixed, paraffin-embedded sections using peptide nucleic acid probes and the catalyzed signal amplification (CSA) technique. Immunogold-silver staining for electron microscopy was performed using anti-ICAM and anti-LFA-1 mouse mAbs. The immunogold particles on epithelial cells of bile ductules and cholangiocytes of CoH cells were counted and analyzed semi-quantitatively. Western blotting was performed to confirm ICAM-1 protein expression. RESULTS: In liver tissues of PBC patients, immunohistochemistry showed aberrant ICAM-1 expression on the plasma membrane of epithelial cells lining bile ductules, and also on mature cholangiocytes but not on hepatocytes in CoH. LFA-1-positive lymphocytes were closely associated with epithelial cells in bile ductules. ICAM 1 expression at protein level was confirmed by Western blot. In situ hybridization demonstrated ICAM-1 mRNA expression in bile ductules and LFA-1 mRNA in lymphocytes infiltrating the bile ductules. By immunoelectron microscopy, ICAM 1 was demonstrated on the basal surface of epithelial cells in bile ductules and on the luminal surfaces of cholangiocytes in damaged CoH. Cells with intermediate morphology resembling progenitor cells in CoH were not labeled with ICAM-1 and LFA-1. CONCLUSION: De novo expression of ICAM-1 both on mature cholangiocytes in CoH and epithelial cells in bile ductules in PBC implies that lymphocyte-induced destruction through adhesion by ICAM-1 and binding of LFA-1-expressing activated lymphocytes takes place not only in bile ductules but also in the CoH. PMID- 16038039 TI - Excretion and detection of SARS coronavirus and its nucleic acid from digestive system. AB - AIM: To study whether severe acute respiratory syndrome coronavirus (SARS-CoV) could be excreted from digestive system. METHODS: Cell culture and semi-nested RT PCR were used to detect SARS-CoV and its RNA from 21 stool and urine samples, and a kind of electropositive filter media particles was used to concentrate the virus in 10 sewage samples from two hospitals receiving SARS patients in Beijing in China. RESULTS: It was demonstrated that there was no live SARS-CoV in all samples collected, but the RNA of SARS-CoV could be detected in seven stool samples from SARS patients with any one of the symptoms of fever, malaise, cough, or dyspnea, in 10 sewage samples before disinfection and 3 samples after disinfection from the two hospitals. The RNA could not be detected in urine and stool samples from patients recovered from SARS. CONCLUSION: Nucleic acid of SARS CoV can be excreted through the stool of patients into sewage system, and the possibility of SARS-CoV transmitting through digestive system cannot be excluded. PMID- 16038040 TI - Effects of iron manipulation on trace elements level in a model of colitis in rats. AB - AIM: Trace elements (TE) metabolism is altered in inflammatory bowel diseases. TE (zinc and copper) are constituents of antioxidant enzymes. Iron is involved in the pathogenesis of chronic inflammation. The aim was to evaluate zinc and copper status and the effects of iron manipulation in experimental colitis. METHODS: Twenty-four male Sprague-Dawley rats were divided into four groups: standard diet, iron-deprived diet, iron-supplemented diet, and sham-treated controls. Macroscopic damage was scored. DNA adducts were measured in the colon. Liver and colonic concentration of TE were measured. RESULTS: Macroscopic damage was reduced in iron-deprived groups and increased in iron-supplemented rats. Damage to the DNA was reduced in iron-deprived groups and increased in iron-supplemented groups. Liver and colonic iron concentrations were reduced in iron-deprived and increased in iron-supplemented rats. Liver zinc concentration was reduced after supplementation whereas colonic levels were similar in controls and treated rats. Liver copper concentration was reduced in all the colitic groups except in the iron-supplemented group whereas colonic concentration was increased in iron deprived rats. CONCLUSION: Iron deprivation diminishes the severity of DNBS colitis while supplementation worsens colitis. Zinc and copper status are modified by iron manipulation. PMID- 16038041 TI - Relationship between beta-catenin expression and epithelial cell proliferation in gastric mucosa with intestinal metaplasia. AB - AIM: To investigate beta-catenin expression in patients with intestinal metaplasia, and to look for a possible relationship between beta-catenin expression and either epithelial proliferation values or Helicobacter pylori (H pylori) infection. METHODS: Twenty patients with complete type intestinal metaplasia were studied. beta-Catenin expression and epithelial cell proliferation in antral mucosa were assessed using an immunohistochemical analysis. H pylori infection was detected by histology and a rapid urease test. RESULTS: Reduced beta-catenin expression on the surface of metaplastic cells was detected in 13 (65%) out of 20 patients. Moreover, in eight (40%) patients intranuclear expression of beta-catenin was found. When patients were analyzed according to H pylori infection, the prevalence of both beta-catenin reduction at the cell surface and its intranuclear localization did not significantly differ between infected and uninfected patients. Cell proliferation was higher in patients with intranuclear beta-catenin expression as compared to the remaining patients, although the difference failed to reach the statistical significance (36+/-8.9 vs 27.2+/-11.4, P = 0.06). On the contrary, a similar cell proliferation value was observed between patients with reduced expression of beta catenin on cell surface and those with a normal expression (28.1+/-11.8 vs 26.1+/ 8.8, P = 0.7). H pylori infection significantly increased cell proliferation (33.3+/-10.2% vs 24.6+/-7.4%, respectively, P = 0.04). CONCLUSION: Both cell surface reduction and intranuclear accumulation of beta-catenin were detected in intestinal metaplasia. The intranuclear localization of beta-catenin increases cell proliferation. H pylori infection does not seem to play a direct role in beta-catenin alterations, whilst it significantly increases cell proliferation. PMID- 16038042 TI - Expression of c-erbB-2 and glutathione S-transferase-pi in hepatocellular carcinoma and its adjacent tissue. AB - AIM: To investigate the possible role of c-erbB-2 and glutathione S-transferase (GST-Pi) in primary hepatocellular carcinogenesis and the relationship between liver hype-rplastic nodule (LHN), liver cirrhosis (LC), and hepatocellular carcinoma (HCC). METHODS: The expression of c-erbB-2 and GST-Pi was detected immunohistochemically in 41 tissue specimens of HCC and 77 specimens of its adjacent tissue. RESULTS: The positive expression of c-erbB-2 in LHN (28.6%) was significantly higher than that in LC (0%) (P = 0.032<0.05), but no significant difference was seen between HCC and LHN or LC (P>0.05, chi2= 0.002, 3.447). The positive expression of GST-Pi in HCC (89.6%) or LHN (71.1%) was significantly higher than that in LC (22.9%, P<0.001, chi2= 49.91, 16.96). There was a significant difference between HCC and LHN (P<0.05, chi2= 6.353). CONCLUSION: The c-erbB-2 expression is an early event in the pathogenesis of HCC. GST-Pi may be a marker enzyme for immunohistochemical detection of human HCC and its preneoplastic lesions. LHN seems to be a preneoplastic lesion related to hepatocarcinogenesis. PMID- 16038043 TI - Association of polymorphisms of IL and CD14 genes with acute severe pancreatitis and septic shock. AB - AIM: To investigate IL-1beta+3 594 in the 5th intron, IL-10-1 082 and CD14-159 polymorphisms in patients with acute pancreatitis (AP) and septic shock. METHODS: The study included 215 patients (109 with acute severe pancreatitis (SAP), 106 with acute mild pancreatitis (MAP)) and 116 healthy volunteers. Genomic DNA was prepared from peripheral blood leukocytes. Genotypes and allele frequencies were determined in patients and healthy controls using restriction fragment length polymorphism analysis of PCR products. RESULTS: The frequencies of IL-1beta+3 594T, IL-10-1082G and CD14-159T allele were similar in patients with mild or severe pancreatitis and in controls. Within SAP patients, no significant differences were found in the allele distribution examined when etiology was studied again. Patients with septic shock showed a significantly higher prevalence of IL-10-1082G allele than those without shock (chi2= 5.921, P = 0.015). CONCLUSION: IL-10-1082G plays an important role in the susceptibility of SAP patients to septic shock. Genetic factors are not important in determination of disease severity or susceptibility to AP. PMID- 16038044 TI - Effects of magnolol and honokiol derived from traditional Chinese herbal remedies on gastrointestinal movement. AB - AIM: To study the effects of magnolol and honokiol on isolated smooth muscle of gastrointestinal tract and their relationship with Ca2+, and on the gastric emptying and the intestinal propulsive activity in mice. METHODS: Routine experimental methods using isolated gastric fundus strips of rats and isolated ileum segments of guinea pigs were adopted to measure the smooth muscle tension. The effects of magnolol 10(-3), 10(-4), 10(-5) mol/L, and honokiol 10(-4), 10( 5), 10(-6) mol/L on the contractility of gastric fundus strips of rats and ileum of guinea pigs induced by acetylcholine (Ach) and 5-hydroxytryptamine (5-HT) was assessed respectively. The method using nuclein and pigment methylene blue was adopted to measure the gastric retention rate of nuclein and the intestinal propulsive ratio of a nutritional semi-solid meal for assessing the effect of magnolol and honokiol (0.5, 2, 20 mg/kg) on gastric emptying and intestinal propulsion. RESULTS: Magnolol and honokiol significantly inhibited the contractility of isolated gastric fundus strips of rats treated with Ach or 5-HT and isolated ileum guinea pigs treated with Ach or CaCl2, and both of them behaved as non-competitive muscarinic antagonists. Magnolol and honokiol inhibited the contraction induced by Ach in Ca2+-free medium and extracellular Ca2+-dependent contraction induced by Ach. Each group of magnolol and honokiol experiments significantly decreased the residual rate of nuclein in the stomach and increased the intestinal propulsive ratio in mice. CONCLUSION: The inhibitory effect of magnolol and honokiol on contractility of the smooth muscles of isolated gastric fundus strips of rats and isolated ileum of guinea pigs is associated with a calcium-antagonistic effect. Magnolol and honokiol can improve the gastric emptying of a semi-solid meal and intestinal propulsive activity in mice. PMID- 16038045 TI - Effect of early nutrition on intestine development of intrauterine growth retardation in rats and its correlation to leptin. AB - AIM: To investigate the intestine and body development of intrauterine growth retardation (IUGR) rats under early different protein diet and to analyze the correlation between leptin and intestine and body development. METHODS: An IUGR rat model was established by food restriction of pregnant female rats. Fifty-six neonatal IUGR rats and 24 neonatal normal rats were randomly divided into normal control group (C group), IUGR model group (SC group), low protein diet IUGR group (SL group), and high protein diet IUGR group (SH group). Eight rats were killed per group at wk 0, 4, and 12. Serum leptin, body weight (BW), body length (BL), intestinal weight (IW), intestinal length (IL), and intestinal disaccharidase (including lactase, maltase, and saccharase) were detected. RESULTS: BW (4.50+/ 0.41 g), BL (5.96+/-0.40 cm), IW (0.05+/-0.01 g), and IL (15.9+/-2.8 cm) in neonatal IUGR rats were much lower than those in C group (6.01+/-0.55 g, 6.26+/ 0.44 cm, 0.10+/-0.02 g, 21.8+/-2.7 cm, P<0.05), while intestinal lactase and maltase activities were higher than those in C group. SH group showed the fastest catch up growth and their BW, BL, IW, and IL reached the C group level at wk 4. SC group showed relatively slower catch up growth than SH group, and their BW, BL, IW did not reach the C group level at wk 4. SL group did not show intestine and body catch up growth. Intestinal maltase (344+/-33 micromol/(min.g)) and saccharase activities (138+/-32 micromol/(min.g)) in SL group were both markedly lower than those in C group (751+/-102, 258+/-27 micromol/(min.g), P<0.05). There were no significant differences in lactase activities at wk 4 and disaccharidase activities at wk 12 among all groups (P>0.05). The leptin level in SL group (0.58+/-0.12 ng/mL) was the highest in all groups, and much lower in SH group (0.21+/-0.03 ng/mL) than that in any other IUGR groups at wk 4 (P<0.05). Leptin was negatively related to BW (r = -0.556, P = 0.001), IW (r = -0.692, P = 0.001) and IL (r = -0.738, P = 0.000) at wk 4, while no correlation was found at wk 12. CONCLUSION: High protein diet is a reasonable early nutritional mode to IUGR rats in promoting intestine and body catch up growth. PMID- 16038046 TI - Protection against hepatic ischemia/reperfusion injury via downregulation of toll like receptor 2 expression by inhibition of Kupffer cell function. AB - AIM: To elucidate the mechanism of liver protection by inhibition of Kupffer cells (KCs) function. METHODS: All the animals were randomly divided into three groups. Blockade group (gadolinium chloride solution (GdCl3) injection plus ischemia/reperfusion (I/R) injury): GdCl3 solution was injected once every 24 h for 2 d via the tail vein before I/R injury. Non-blockade group (saline solution injection plus I/R injury): saline instead of GdCl3 as a control was injected as in the blockade group. Sham group: saline was injected without I/R injury. Liver samples were collected 4 h after blood inflow restoration. The blockade of the function of KCs was verified by immunostaining with an anti-CD68 mAb. Toll-like receptor 2 (TLR2) was immunostained with a goat antimouse polyclonal anti-TLR2 antibody. Membrane proteins were extracted from the liver samples and TLR2 protein was analyzed by Western blot. Portal vein serum and plasma were taken respectively at the same time point for further detection of the levels of tumor necrosis factor-alpha (TNF-alpha) and alanine aminotransferase (ALT), an indicator of liver function. RESULTS: Compared to non-blockade group, CD68+ cells significantly reduced in blockade group (OPTDI, optical density integral): 32.97+/-10.55 vs 185.65+/-21.88, P<0.01) and the liver function impairment was relieved partially (level of ALT: 435.89+/-178.37 U/L vs 890.21+/-272.91 U/L, P<0.01). The expression of TLR2 protein in blockade group significantly decreased compared to that in non-blockade group (method of immunohistochemistry, OPDTI: 75.74+/-17.44 vs 170.58+/-25.14, P<0.01; method of Western blot, A value: 125.89+/-15.49 vs 433.91+/-35.53, P<0.01). The latter correlated with the variation of CD68 staining (r = 0.745, P<0.05). Also the level of portal vein TNF alpha decreased in blockade group compared to that in non-blockade group (84.45+/ 14.73 ng/L vs 112.32+/-17.56 ng/L, P<0.05), but was still higher than that in sham group (84.45+/-14.73 ng/L vs 6.07+/-5.33 ng/L, P<0.01). CONCLUSION: Inhibition of the function of KCs may protect liver against I/R injury via downregulation of the expression of TLR2. PMID- 16038047 TI - Effect of c-fos antisense probe on prostaglandin E2-induced upregulation of vascular endothelial growth factor mRNA in human liver cancer cells. AB - AIM: To examine the effect of prostaglandin E2 (PGE2) on the expression of vascular endothelial growth factor (VEGF) mRNA in the human hepatocellular carcinoma (HCC) HepG2 cells and the possible involvement of c-fos protein in this process. METHODS: Human HCC HepG2 cells were divided into three groups treated respectively with PGE2, a combination of PGE2 and c-fos antisense oligodeoxynucleotide (ASO), and PGE2 plus c-fos sense oligodeoxynucleotide (SO). The expression of VEGF mRNA in HepG2 cells after different treatments was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). The relative expression level of VEGF mRNA in HepG2 cells in each group was measured. RESULTS: Administration of PGE2 resulted in an increased expression of c-fos and VEGF mRNA in HepG2 cells. The relative expression level of c-fos mRNA reached the peak at 3 h (68.4+/-4.7%) after PGE2 treatment, which was significantly higher than that at 0 h (20.6+/-1.7%, P<0.01). Whereas, the highest expression level of VEGF mRNA was observed at 6 h (100.5+/-6.1%) after PGE2 treatment, which was significantly higher than that at 0 h (33.2+/-2.4%, P<0.01). C-fos ASO significantly reduced PGE2-induced VEGF mRNA expression in HepG2 cells. CONCLUSION: PGE2 increases the expression and secretion of VEGF in HCC cells by activating the transcription factor c-fos, promotes the angiogenesis of HCC and plays an important role in the pathogenesis of liver cancer. PMID- 16038048 TI - Risk factors for primary liver carcinoma in Chinese population. AB - AIM: To evaluate the risk factors for primary liver carcinoma (PLC) in Chinese population. METHODS: Chinese Biomedical Literature Database, China Hospital Knowledge Database and MEDLINE were searched. All the related literatures were screened, and the risk factors for PLC in Chinese population were studied. Heterogeneity was evaluated by odds ratio (OR) q test. Combined OR and its 95% confidence interval (95%CI) were calculated, the association between the investigated risk factors and PLC was determined. Validity and bias of the findings were evaluated by sensitivity analysis and funnel plot analysis respectively. RESULTS: Fifty-five of one hundred and ninety identified studies were accepted according to the inclusive criteria. Ten factors related to PLC were demonstrated by sensitive analysis and funnel plot analysis. They were cirrhosis (OR = 11.97, P = 0.000), HBV infection (OR = 11.34, P = 0.000), HCV infection (OR = 4.28, P = 0.000), family history of liver cancer (OR = 3.49, P = 0.000), unstable emotion (OR = 2.20, P = 0.000), depressed characters (OR = 3.07, P = 0.000), aflatoxin (OR = 1.80, P = 0.000), alcoholic (OR = 1.88, P = 0.000), intake of musty food (OR = 1.87, P = 0.000) and drinking contaminated water from pond (OR = 1.77, P = 0.003). CONCLUSION: The main risk factors for PLC in China are liver diseases, family history of liver carcinoma, poor psychic status, aflatoxin, and some unhealthy behaviors. PMID- 16038049 TI - Effects of sulfasalazine on biopsy mucosal pathologies and histological grading of patients with active ulcerative colitis. AB - AIM: To investigate the mechanisms of sulfasalazine (SASP) in the treatment of ulcerative colitis (UC). METHODS: Changes of pathological signs and histological grading of 106 patients with active UC were observed before and after the treatment with SASP, 1 g, thrice daily for 6 wk. RESULTS: The effect of SASP on the vasculitis in lamina propria was 48.2% and 17.4% in the mild active UC (P<0.001) and 68% and 26.7% in the moderate active UC (P<0.001) before and after treatment. Fibroid necrosis of vessel wall was found in one case of mild UC and two cases of moderate UC before treatment and was not found after treatment. No thrombosis was found in mild UC before and after treatment, while thrombosis was found in one case of moderate UC before treatment. The effect on mucosal glandular abnormality was 30.4% and 13.0% in mild UC (P<0.05), and 42% and 40% in moderate UC (P>0.05) before and after treatment. The rate of eosinophil infiltration was 98.2% and 80.4% in mild UC (P<0.01), and 100% and 91.1% in moderate UC (P<0.05) before and after treatment. The effect on crypt abscess was 21.4% and 4.4% in mild UC (P<0.05), and 48% and 13.3% in moderate UC (P<0.001) before and after treatment. The effect on mucosal pathohistological grading was 2.00+/-0.84 and 0.91+/-0.46 in mild UC (P<0.001), and 2.49+/-0.84 and 1.31+/-0.75 in moderate UC (P<0.001) before and after treatment. CONCLUSION: SASP can improve small vessel lesions and crypt abscesses and reduce neutrophilic and eosinophilic leukocyte infiltration in inflammatory mucosa of UC. PMID- 16038050 TI - Liposome transfected to plasmid-encoding endostatin gene combined with radiotherapy inhibits liver cancer growth in nude mice. AB - AIM: To evaluate whether intratumoral injection of liposome-endostatin complexes could enhance the antitumor efficacy of radiation therapy in human liver carcinoma (BEL7402) model. METHODS: Recombinant plasmid pcDNA3.End was transfected into human liver carcinoma cell line (BEL7402) with lipofectamine to produce conditioned medium. Then BEL7402 cells and human umbilical vein endothelial cells (HUVECs) were treated with the conditioned medium. Cell cycle and apoptosis were analyzed by flow cytometer and endothelial cell proliferation rates were determined by MTT assay. The antitumor efficacy of endostatin gene combined with ionizing radiation in mouse xenograft liver tumor was observed. RESULTS: Endostatin significantly suppressed the S phase fraction and increased the apoptotic index in HUVECs. In contrast, endostatin treatment had no effect on BEL7402 cell apoptosis (2.1+/-0.3% vs 8.9+/-1.3%, t = 8.83, P = 0.009<0.01) or cell cycle distribution (17.2+/-2.3% vs 9.8+/-1.2%, t = 4.94, P = 0.016<0.05). The MTT assay showed that endostatin significantly inhibited the proliferation of HUVECs by 46.4%. The combination of local endostatin gene therapy with radiation therapy significantly inhibited the growth of human liver carcinoma BEL7402 xenografts, the inhibition rate of tumor size was 69.8% on d 28 compared to the untreated group. The tumor volume in the pcDNA3.End combined with radiation therapy group (249+/-83 mm3) was significantly different from that in the untreated group (823+/-148 mm3, t = 5.86, P = 0.009<0.01) or in the pcDNA3 group (717+/-94 mm3, t = 6.46, P = 0.003<0.01). Endostatin or the radiation alone also inhibited the growth of liver tumor in vivo, but their inhibition effects were weaker than those of endostatin combined with radiation, the inhibition rates on d 28 were 44.7% and 40.1%, respectively. CONCLUSION: Endostatin not only significantly suppresses tumor growth but also enhances the antitumor efficacy of radiation therapy in human carcinoma xenograft. PMID- 16038051 TI - Small bowel non-Hodgkin's lymphoma remaining in complete remission by surgical resection and adjuvant rituximab therapy. AB - A 44-year-old man was referred to our hospital with intermittent abdominal pain. Because distention of fluid- and gas-filled loops of small intestine was proved by X-ray, the patient was diagnosed as having small bowel obstruction. A laparotomy revealed a segmental stenosis in the jejunum, which showed diffuse thickening of the intestinal wall. Some mesenteric lymph nodes were swollen. Pathological examination was defined. We diagnosed diffuse large B-cell lymphoma based on the pathological findings of diffuse transmural infiltration of large lymphoid cells and flow-cytometric analyses. Rituximab was administered as adjuvant therapy at weekly doses of 375 mg/m2. Four cycles were performed every 6 mo and he remained CR. Rituximab may be effective as adjuvant therapy. PMID- 16038052 TI - Ovarian carcinoma in two patients with chronic liver disease. AB - Ascites is a common and debilitating complication of cirrhosis. However, patients with chronic liver disease are not spared from other causes of ascites and physicians should be careful not to miss an underlying malignancy. Ovarian cancer is an insidious disease, which is difficult to diagnose and it ranks first in mortality among all gynecological cancers. Here, we present two cases of patients with chronic liver disease that developed ascites not simply because of cirrhosis but as a manifestation of ovarian cancer. We would like to emphasize that the causes of ascites, other than the liver itself, should not be overlooked in patients with chronic liver disease. PMID- 16038053 TI - Acute pancreatitis caused by leptospirosis: report of two cases. AB - Two cases of acute pancreatitis with leptospirosis are reported in this article. CASE 1: A 68-year-old woman, presented initially with abdominal pain, nausea, vomiting, and jaundice. She was in poor general condition, and had acute abdominal signs and symptoms on physical examination. Emergency laparotomy was performed, acute pancreatitis and leptospirosis were diagnosed on the basis of surgical findings and serological tests. The patient died on postoperative d 6. CASE 2: A 62-year-old man, presented with fever, jaundice, nausea, vomiting, and malaise. Acute pancreatitis associated with leptospirosis was diagnosed, according to abdominal CT scanning and serological tests. The patient recovered fully with antibiotic treatment and nutritional support within 19 d. PMID- 16038054 TI - The plant Golgi apparatus--going with the flow. AB - The plant Golgi apparatus is composed of many separate stacks of cisternae which are often associated with the endoplasmic reticulum and which in many cell types are motile. In this review, we discuss the latest data on the molecular regulation of Golgi function. The concept of the Golgi as a distinct organelle is challenged and the possibility of a continuum between the endoplasmic reticulum and Golgi is proposed. PMID- 16038055 TI - Golgi reassembly after mitosis: the AAA family meets the ubiquitin family. AB - The Golgi apparatus in animal cells breaks down at the onset of mitosis and is later rebuilt in the two daughter cells. Two AAA ATPases, NSF and p97/VCP, have been implicated in regulating membrane fusion steps that lead to regrowth of Golgi cisternae from mitotic fragments. NSF dissociates complexes of SNARE proteins, thereby reactivating them to mediate membrane fusion. However, NSF has a second function in regulating SNARE pairing together with the ubiquitin-like protein GATE-16. p97/VCP, on the other hand, is involved in a cycle of ubiquitination and deubiquitination of an unknown target that governs Golgi membrane dynamics. Here, these findings are reviewed and discussed in the context of the increasingly evident role of ubiquitin in membrane traffic processes. PMID- 16038056 TI - SNAREs and traffic. AB - SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) are now generally accepted to be the major players in the final stage of the docking and the subsequent fusion of diverse vesicle-mediated transport events. The SNARE-mediated process is conserved evolutionally from yeast to human, as well as mechanistically and structurally across different transport events in eukaryotic cells. In the post-genomic era, a fairly complete list of "all" SNAREs in several organisms (including human) can now be made. This review aims to summarize the key properties and the mechanism of action of SNAREs in mammalian cells. PMID- 16038057 TI - Familial cancer and ARLTS1. PMID- 16038058 TI - Thrombosis of the cerebral veins and sinuses. PMID- 16038059 TI - Thrombosis of the cerebral veins and sinuses. PMID- 16038060 TI - The costs of institutional review boards. PMID- 16038061 TI - Learning to mentor. PMID- 16038063 TI - Bibliography. Current world literature. Cardiopulmonary monitoring. PMID- 16038062 TI - The role of laparoscopy in establishing diaphragmatic injury in lower chest wounds. PMID- 16038064 TI - Response to the letter on bullying. PMID- 16038065 TI - Bullying. PMID- 16038067 TI - [Regional meeting of the Japanese Association of Anatomists. Japan. 2004. Abstracts]. PMID- 16038066 TI - Interactive visual and postvisual processes and their roles in form-specific memory. AB - Effects of depth of encoding on form-specific memory were examined. After viewing words (e.g., "bear") presented centrally during initial encoding, participants completed word stems (e.g., "BEA") presented laterally and pattern masked during subsequent test. When the encoding task was perceptual, letter-case specific memory was not observed, unlike in previous experiments without pattern masking. However, when the encoding task required both perceptual and conceptual processing, letter-case specific memory was observed in direct right-hemisphere, but not in direct left-hemisphere, test presentations, like in previous studies without pattern masking. Results were not influenced by whether stems were completed to form the first words that came to mind or words explicitly retrieved from encoding. Depth of encoding may influence form-specific memory through interactive processing of visual and postvisual information. PMID- 16038068 TI - [113th Scientific meeting of the Japan Prosthodontic Society. Osaka, Japan. May 14-15, 2005. Program and Abstracts]. PMID- 16038069 TI - Cytoskeletal interactions with the outside world. AB - Cell motility depends on the conversion of extracellular cues into intracellular cytoskeletal responses. A recent Keystone meeting on cell migration and adhesion showcased advances in the field. PMID- 16038070 TI - Acquired immune deficiency syndrome and infection with human immunodeficiency virus. PMID- 16038071 TI - The Cruzan decision: 9.5 theses for discussion. PMID- 16038072 TI - Nancy Cruzan and the best interest standard. PMID- 16038073 TI - Advance directives in the 1990s. PMID- 16038074 TI - Nancy Cruzan and the "right to die"--a Jewish perspective. PMID- 16038076 TI - Clear and convincing evidence: the case of Nancy Cruzan. PMID- 16038075 TI - The constitution and the right to die. PMID- 16038077 TI - Genetic testing: for whose benefit is it provided? PMID- 16038078 TI - Ethical conflicts in health care technology. PMID- 16038079 TI - Ethical issues in genetic engineering: a survey. PMID- 16038080 TI - Telling illness: the patient-physician story. PMID- 16038081 TI - The Human Genome Project: what questions does it raise for theology and ethics? PMID- 16038082 TI - The human significance of the genome project. PMID- 16038083 TI - Does Jewish law require us to tell patients the truth about their terminal conditions? PMID- 16038084 TI - Challenge for fair trade. PMID- 16038085 TI - Weary father left to count the days: doctors hope technology can sustain fetus. PMID- 16038086 TI - BMA drops its opposition to doctor-assisted suicide. PMID- 16038087 TI - Improper insulin administration may explain poor diabetes control. PMID- 16038088 TI - The yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe: models for cell biology research. AB - Yeast species provide excellent models for fundamental biological research. In this review, I will describe characteristics of the two most common laboratory systems: the fission yeast Schizosaccharomyces pombe, and the budding yeast Saccharomyces cerevisiae. They have substantial similarities that make them powerful as research tools, and also striking biological differences that make them complementary experimental models. Each provides unique tools for understanding environmental effects on cellular systems. PMID- 16038089 TI - Worms in space? A model biological dosimeter. AB - Although it is well known that radiation causes mutational damage, little is known about the biological effects of long-term exposure to radiation in space. Exposure to radiation can result in serious heritable defects in experimental animals, and in humans, susceptibility to cancer, radiation-sickness, and death at high dosages. It is possible to do ground controlled studies of different types of radiation on experimental animals and to physically measure radiation on the space station or on space probes. However, the actual biological affects of long-term exposure to the full range of space radiation have not been studied, and little information is available about the biological consequences of solar flares. Biological systems are not simply passive recording instruments. They respond differently under different conditions, and thus it is important to be able to collect data from a living animal. There are technical difficulties that restrict the placement of an experimental organism in a space environment for long periods of time, in a manner that allows for the recovery of genetic data. Use of the self-fertilizing hermaphroditic nematode, Caenorhabditis elegans offers potential for the design of a biological dosimeter. In this paper, we describe the advantages of this model system and review the literature of C. elegans in space. PMID- 16038090 TI - Drosophila melanogaster--the model organism of choice for the complex biology of multi-cellular organisms. AB - Drosophila melanogaster has been intensely studied for almost 100 years. The sophisticated array of genetic and molecular tools that have evolved for analysis of gene function in this organism are unique. Further, Drosophila is a complex multi-cellular organism in which many aspects of development and behavior parallel those in human beings. These combined advantages have permitted research in Drosophila to make seminal contributions to the understanding of fundamental biological processes and ensure that Drosophila will continue to provide unique insights in the genomic era. An overview of the genetic methodologies available in Drosophila is given here, together with examples of outstanding recent contributions of Drosophila to our understanding of cell and organismal biology. The growing contribution of Drosophila to our knowledge of gravity-related responses is addressed. PMID- 16038091 TI - Use of animal models for space flight physiology studies, with special focus on the immune system. AB - Animal models have been used to study the effects of space flight on physiological systems. The animal models have been used because of the limited availability of human subjects for studies to be carried out in space as well as because of the need to carry out experiments requiring samples and experimental conditions that cannot be performed using humans. Experiments have been carried out in space using a variety of species, and included developmental biology studies. These species included rats, mice, non-human primates, fish, invertebrates, amphibians and insects. The species were chosen because they best fit the experimental conditions required for the experiments. Experiments with animals have also been carried out utilizing ground-based models that simulate some of the effects of exposure to space flight conditions. Most of the animal studies have generated results that parallel the effects of space flight on human physiological systems. Systems studied have included the neurovestibular system, the musculoskeletal system, the immune system, the neurological system, the hematological system, and the cardiovascular system. Hindlimb unloading, a ground based model of some of the effects of space flight on the immune system, has been used to study the effects of space flight conditions on physiological parameters. For the immune system, exposure to hindlimb unloading has been shown to results in alterations of the immune system similar to those observed after space flight. This has permitted the development of experiments that demonstrated compromised resistance to infection in rodents maintained in the hindlimb unloading model as well as the beginning of studies to develop countermeasures to ameliorate or prevent such occurrences. Although there are limitations to the use of animal models for the effects of space flight on physiological systems, the animal models should prove very valuable in designing countermeasures for exploration class missions of the future. PMID- 16038092 TI - Exercise and pharmacological countermeasures for bone loss during long-duration space flight. AB - Bone loss in the lower extremities and lumbar spine is an established consequence of long-duration human space flight. Astronauts typically lose as much bone mass in the proximal femur in 1 month as postmenopausal women on Earth lose in 1 year. Pharmacological interventions have not been routinely used in space, and countermeasure programs have depended solely upon exercise. However, it is clear that the osteogenic stimulus from exercise has been inadequate to maintain bone mass, due to insufficient load or duration. Attention has therefore been focused on several pharmacological interventions that have been successful in preventing or attenuating osteoporosis on Earth. Anti-resorptives are the class of drugs most commonly used to treat osteoporosis in postmenopausal women, notably alendronate sodium, risedronate sodium, zoledronic acid, and selective estrogen receptor modulators, such as raloxifene. There has also been considerable recent interest in anabolic agents such as parathyroid hormone (PTH) and teriparatide (rhPTH [1-34]). Vitamin D and calcium supplementation have also been used. Recent studies of kindreds with abnormally high bone mineral density have provided insight into the genetic regulation of bone mass. This has led to potential therapeutic interventions based on the LRP5, Wnt and BMP2 pathways. Another target is the RANK-L/osteoprotegerin signaling pathway, which influences bone turnover by regulating osteoclast formation and maturation. Trials using such therapies in space are being planned. Among the factors to be considered are dose response relationships, bone quality, post-use recovery, and combination therapies--all of which may have unique characteristics when the drugs are used in space. PMID- 16038093 TI - Consequences of cardiovascular adaptation to spaceflight: implications for the use of pharmacological countermeasures. AB - There is little evidence obtained from space flight to support the notion that occurrence of cardiac dysrhythmias, impaired cardiac and vascular function, and manifestation of asymptomatic cardiovascular disease represent serious risks during space flight. Therefore, the development of orthostatic hypotension and instability immediately after return from spaceflight probably reflect the most significant operational risks associated with the cardiovascular system of astronauts. Significant reductions in stroke volume and lower reserve for increasing peripheral vascular resistance contribute to ineffective maintenance of systemic arterial blood pressure during standing after spaceflight despite compensatory elevations in heart rate. The primary mechanism underlying reduced stroke volume appears to be a reduction in preload associated with less circulating blood volume while inadequate peripheral vasoconstriction may be caused partly by hyporeactivity of receptors that control arterial smooth muscle function. A focus for development of future countermeasures for hemodynamic responses to central hypovolemia includes the potential application of pharmacological agents that specifically target and restore blood volume (e.g., fludrocortisone, electrolyte-containing beverages) and reserve for vasoconstriction (e.g., midodrine, vasopressin). Based on systematic evaluations, acute physical exercise designed to elicit maximal effort or inspiratory resistance have shown promise as successful countermeasures that provide protection against development of orthostatic hypotension and intolerance without potential risks and side effects associated with specific pharmacological interventions. PMID- 16038095 TI - Evaluation of a silane quaternary ammonium salt as an antimicrobial surface treatment. PMID- 16038094 TI - Diet as a factor in behavioral radiation protection following exposure to heavy particles. AB - Major risks associated with radiation exposures on deep space missions include carcinogenesis due to heavy-particle exposure of cancer-prone tissues and performance decrements due to neurological damage produced by heavy particles. Because exposure to heavy particles can cause oxidative stress, it is possible that antioxidants can be used to mitigate these risks (and possibly some health risks of microgravity). To assess the capacity of antioxidant diets to mitigate the effects of exposure to heavy particles, rats were maintained on antioxidant diets containing 2% blueberry or strawberry extract or a control diet for 8 weeks prior to exposure to 1.5 or 2.0 Gy of accelerated iron particles at Brookhaven National Laboratory. Following irradiation rats were tested on a series of behavioral tasks: amphetamine-induced taste aversion learning, operant responding and spatial learning and memory. The results indicated that the performance of the irradiated rats maintained on the antioxidant diets was, in general, significantly better than that of the control animals, although the effectiveness of the diets ameliorating the radiation-induced deterioration in performance varied as a function of both the specific diet and the specific endpoint. In addition, animals fed antioxidant diets prior to exposure showed reduced heavy particle-induced tumorigenesis one year after exposure compared to the animals fed the control diet. These results suggest that antioxidant diets have the potential to serve as part of a system designed to provide protection to astronauts against the effects of heavy particles on exploratory missions outside the magnetic field of the earth. PMID- 16038096 TI - Screening and identification of cryopreservative agents for human cellular biotechnology experiments in microgravity. PMID- 16038097 TI - Pseudomonas aeruginosa growth and production of Exotoxin A in static and modeled microgravity environments. PMID- 16038098 TI - Development and testing of an efficient LED intracanopy lighting design for minimizing Equivalent System Mass in an advanced life-support system. PMID- 16038099 TI - Cell behavior in simulated microgravity: a comparison of results obtained with RWV and RPM. PMID- 16038100 TI - Methods for the culture of C. elegans and S. cerevisiae in microgravity. AB - To support the study of the effects of microgravity on biological systems, our group is developing and testing methods that allow the cultivation of C. elegans and S. cerevisiae in microgravity. Our aim is to develop the experimental means by which investigators may conduct peer reviewed biological experiments with C. elegans or S. cerevisiae in microgravity. Our protocols are aimed at enabling investigators to grow these organisms for extended periods during which samples may be sub-cultured, collected, preserved, frozen, and/or returned to earth for analysis. Data presented include characterization of the growth phenotype of these organisms in liquid medium in OptiCells(TM) (Biocrystal, LTD). PMID- 16038101 TI - Development of the EMCS hardware for multigenerational growth of Drosophila melanogaster in space. PMID- 16038102 TI - Neurophysiological long-term recordings in space: experiments Scorpi and Scorpi T. PMID- 16038104 TI - Environmental factors and lupus: are we looking too late? PMID- 16038103 TI - A Toll for lupus. AB - Toll-like receptor (TLR)-9 recognizes CpG motifs in microbial DNA. TLR9 signalling stimulates innate antimicrobial immunity and modulates adaptive immune responses including autoimmunity against chromatin, e.g., in systemic lupus erythematosus (SLE). This review summarizes the available data for a role of TLR9 signalling in lupus and discusses the following questions that arise from these observations: 1) Is CpG-DNA/TLR9 interaction involved in infection-induced disease activity of lupus? 2) What are the risks of CpG motifs in vaccine adjuvants for lupus patients? 3) Is TLR9 signalling involved in the pathogenesis of lupus by recognizing self DNA? PMID- 16038105 TI - Correlation of membranous glomerular ultrastructural changes with disease severity and outcome in lupus patients initiating cyclophosphamide therapy. AB - The aim of this study was to assess the utility applying an electron microscopy (EM) scoring system used in idiopathic membranous nephritis based on the location of subepithelial and/or intramembranous electron dense deposits in interpretation of renal biopsies from patients with lupus nephritis. We selected patients with electron dense deposits traditionally associated with membranous changes on EM from 84 patients treated with bolus cyclophosphamide, with five years follow-up. An EM scoring system designed for idiopathic membranous nephritis was applied (stages I or II, mild changes; stages III or IV, advanced changes). Twenty-seven out of 84 had membranous changes by light microscopy, of whom 22 had satisfactory tissue for EM membranous analysis. Eleven out of 22 had mild EM changes (EM stage I or II); 11 had advanced disease (EM stage III). Advanced EM stage was associated with a higher serum creatinine at entry when tests were adjusted for WHO class (2.62 +/- 0.6 versus 1.31 +/- 0.28 mg/dL, P < 0.022 by ANOVA), and EM stage was independent of NIH activity or chronicity indexes or disease duration. After five years, adverse outcomes (death or dialysis) were seen in one of the 11 patients with EM stages I-II versus five of the 11 EM stage III patients (P < 0.07). Advanced membranous type electron dense deposition in lupus as assessed by EM was associated with worse renal function in patients with comparable WHO classification and NIH activity and chronicity indexes. In this group of lupus patients initiating cyclophosphamide for severe nephritis, EM stage provided important additional information regarding the extent of renal injury. PMID- 16038106 TI - Long-term thalidomide use in refractory cutaneous lesions of lupus erythematosus: a 65 series of Brazilian patients. AB - Thalidomide has been reported as efficacious in refractory cutaneous lupus erythematosus (LE). The most fearful side-effects are teratogenicity and neuropathy. We reported clinical efficacy of long-term low-dose use of thalidomide in 65 patients with LE, emphasizing the prevalence of adverse effects, especially of neuropathy and its related factors. Data obtained from medical records included age, sex, disease duration, and the presence of diagnostic criteria for systemic lupus erythematosus (SLE), the extent and activity of cutaneous lesions and previous treatments. Sixty-three patients (98.9%) presented complete or partial improvement with thalidomide therapy. Drowsiness occurred in 50 patients (77%). Twenty-eight patients (43.2%) presented neuropathy symptoms. Nerve conduction studies were done in 21 (75%) of them and were abnormal in 12 (57%). With the interruption of thalidomide, 24 (82.5%) had complete or partial improvement of neuropathy symptoms and 23 (82%) of them had cutaneous relapse. There were no significant differences between those who developed or not neuropathy in treatment duration, age, total dose and systemic versus cutaneous LE. In conclusion, thalidomide can be used in refractory cutaneous LE with great efficacy and relative security. Controlled studies with schemes with lower doses or intermittent usage or alternative drugs are wanted to reduce the burden of cutaneous morbidity of lupus erythematosus. PMID- 16038107 TI - Association between beta2-glycoprotein I gene polymorphisms and pediatric SLE and antiphospholipid antibodies. AB - Antibodies against phospholipids (PL) and PL-binding proteins have been causally implicated in antiphospholipid syndrome (APS). Mutations in the fifth domain of the beta2-glycoprotein I (beta2GPI) protein, a putative PL-binding site, may play a critical role in APS pathogenesis. The purpose of this study was to identify associations between beta2GPI mutations and both antiphospholipid antibodies (aPL) and their associated clinical manifestations in a pediatric and adolescent cohort and to search for novel mutations. Genetic analysis of beta2GPI was performed in 58 youths with systemic lupus erythematosus (SLE) and/or aPL, to identify known polymorphisms at amino acids 247 and 306 as well as novel mutations in exon 7 of the beta2GPI gene, and their association with aPL associated clinical manifestations. Our results demonstrate an association between substitution of Val for Leu at AA247 (L247V) of beta2GPI and both the development of aPL (P = 0.05) and aPL-associated clinical manifestations (P = 0.03) among pediatric patients. The odds ratio associated with risk of aPL associated clinical manifestations for the homozygous VV polymorphism was 5.5 (CI 1.3-23, P = 0.03) for the overall cohort, and 4.75 (CI 0.66-55.49, P = 0.06) after adjusting for ethnicity. The association was not significant after stratifying for SLE versus non-SLE. Association between the VV genotype at amino acid 247 of beta2GPI and clinical disease supports a genetic cause for APS among children and adolescents. Neither novel exon 7 beta2GPI mutations or the previously described C306G polymorphism was identified in this pediatric cohort. PMID- 16038108 TI - Evaluation of average amount of cerebral blood flow measured by brain perfusion index in patients with neuropsychiatric systemic lupus erythematosus. AB - We used the brain perfusion index (BPI), an indicator of the average amount of cerebral blood flow (CBF), to evaluate the usefulness of the average amount of CBF for neuropsychiatric systemic lupus erythematosus (NPSLE). Of the seventy three SLE patients examined in this study (total 100 scans), 16 patients (23 scans) had already been diagnosed with NPSLE based on clinical symptoms indicative of central nervous system involvement. In addition, 12 patients (17 scans) exhibited the antiphospholipid antibody syndrome (APS). BPI is significantly influenced by age and we therefore used the BPI ratio (ratio of age predicted BPI to measured BPI value) for each assessment. The mean BPI value of 100 scans was 11.2 +/- 2.79, and the mean BPI ratio was 0.99 +/- 0.24 in all SLE patients. The mean BPI ratio among NPSLE (0.84 +/- 0.19) was significantly lower than that of the non-NPSLE patients (1.04 +/- 0.24) (P < 0.0005). However, there was no difference in the mean BPI ratio between APS patients (0.98 +/- 0.24) and non-APS patients (0.99 +/- 0.25). These results indicate that the mean CBF assessed by the BPI ratio using SPECT is of use in the evaluation of central nervous system involvement in SLE patients. PMID- 16038109 TI - Large pericardial effusions due to systemic lupus erythematosus: a report of eight cases. AB - The aim of this study was to describe the clinical, echocardiographic and laboratory characteristics of large pericardial effusions and cardiac tamponade secondary to systemic lupus erythematosus (SLE). An ongoing prospective study was conducted at Tygerberg Academic Hospital, South Africa between 1996 and 2002. All patients older than 13 years presenting with large pericardial effusions (> 10 mm) requiring pericardiocentesis were included. Eight cases (out of 258) were diagnosed with SLE. The mean (SD) age was 29.5 (10.7) years. Common clinical features were Raynaud's phenomenon, arthralgia and lupus nephritis class III/IV. Echocardiography showed Libman-Sacks endocarditis (LSE) in all the mitral valves. Two patients developed transient left ventricular dysfunction; both these patients had pancarditis. Typical serological findings included antinuclear antibodies, anti-double stranded DNA antibodies, low complement C4 levels and low C3 levels. CRP was elevated in six cases. Treatment consisted of oral steroids and complete drainage of the pericardial effusions. No repeat pericardial effusions or constrictive pericarditis developed amongst the survivors (3.1 years follow up). This study concludes that large pericardial effusions due to SLE are rare, and associated with nephritis, LSE and myocardial dysfunction. Treatment with steroids and complete drainage is associated with a good cardiac outcome. PMID- 16038110 TI - Hypercatabolism of IgG in mice with lupus-like syndrome. AB - The metabolism of radioiodinated mouse IgG was studied in mice with lupus-like syndrome before and after the onset of the disease. Before the onset of the disease, the pharmacokinetic parameters of IgG in MLR-1pr and Pristane-primed Balb/c mice were within the normal range of values. After the onset of the disease a considerable increase in the catabolic rate of IgG was recorded abbreviating its half life to less than one third of the normal value. The increased catabolism of IgG could not be related to the concentration--catabolism effect or to the presence of rheumatoid factor and autoantibody or to the IgG loss through the kidney and gastrointestinal tract. The hypercatabolism of IgG was explained by disease-induced impairment of the function of the receptor FcRn, which regulates the homeostasis of IgG. PMID- 16038111 TI - Review of presentations at the 6th European Lupus Meeting 3-5 March 2005. AB - The 6th European Lupus Meeting was held at the Royal College of Physicians of London and was attended by 450 delegates. The conference brought together leading speakers from Europe and North America who reviewed current knowledge and exciting new developments in both clinical and basic science aspects of systemic lupus erythematosus. This review summarizes the major points covered in each session. PMID- 16038112 TI - Spontaneous splenic artery hemorrhage with secondary antiphospholipid syndrome in lupus: a case report. AB - Spontaneous hemorrhage is a rare complication of lupus. We describe a 36-year old female with lupus who suffered spontaneous, nontraumatic hemorrhage from branches of the splenic artery with massive blood loss while being treated for a lupus flare. We compare this to the two other reported cases of similar lupus associated splenic artery hemorrhage documented in the literature, both of which had significant pre-existing hemorrhagic risk factors. Spontaneous, nontraumatic hemorrhage of the splenic artery in the absence of risk factors, and in a patient with secondary antiphospholipid syndrome, has been previously undescribed in lupus. PMID- 16038113 TI - Systemic lupus erythematosus with refractory hemolytic anemia effectively treated with cyclosporin A: a case report. AB - A 51 year old woman with systemic lupus erythematosus (SLE) serially developed thrombocytopenia, arthritis, lupus nephritis, pleuritis, mesenteric vasculitis and refractory hemolytic anemia during the past 19 years prior to presentation. The woman had been managed with high doses of prednisolone, splenectomy, methylprednisolone pulse therapy and cytotoxic drugs, including oral cyclophosphamide, azathioprine, mexotrexate and monthly parenteral cyclophosphamide for hemolytic anemia. After two months of therapeutic trial with Cyclosporin A (CsA) (3 mg/kg), the follow-up hemoglobin (Hb) level was increased to 12.0 gm% and the dose ofprednislone was reduced to 5 mg every other day without occurring rebound during the subsequent three months. There were no obvious side effects from the medication. PMID- 16038114 TI - Anti-glutamic acid decarboxylase antibodies in a patient with systemic lupus erythematosus and fibromyalgia symptoms. AB - We report the case of a 29-year old female nurse with a five-year history of systemic lupus erythematosus (SLE) involving multiple systems and on chronic prednisone therapy. This patient has a coexisting diagnosis of fibromyalgia fulfilling ACR criteria. A recent deterioration in her level of functioning in addition to a flare of her inflammatory disease led to further evaluation. During the course of investigation an anti-glutamic acid decarboxylase antibody was found to be present and significantly elevated. A therapeutic trial of baclofen did result in improvement of her subjective myalgias. We raise the possibility of an autoimmune contribution to myalgic symptoms in a portion of SLE patients. PMID- 16038115 TI - Urticarial vasculitis and subacute cutaneous lupus erythematosus. PMID- 16038116 TI - Azathioprine induced hepatic veno-occlusive disease in systemic lupus erythematosus. PMID- 16038117 TI - Systemic lupus erythematosus induced by therapy with interferon-beta in a patient with multiple sclerosis. PMID- 16038118 TI - [Early and intensive insulin therapy lowers late complications]. PMID- 16038119 TI - [Rasagiline. A new monoamine oxidase b inhibitor for Parkinson treatment]. PMID- 16038120 TI - [Pharmaceutical care of Parkinson patients]. PMID- 16038121 TI - [Acetylsalicylic acid and vitamin C]. PMID- 16038122 TI - [How can we make drug taking easier for children?]. PMID- 16038123 TI - [Advanced course of Parkinson disease. Little improved motion with continuous intestinal levodopa administration]. PMID- 16038124 TI - [Pathogenesis and therapeutic strategy]. PMID- 16038125 TI - [Better transplantation survival with aspirin?]. PMID- 16038126 TI - [Prevention of stroke with aspirin, vitamin E is ineffective]. PMID- 16038127 TI - [Hyperhomocysteinemia. Causal risk factors for stroke]. PMID- 16038128 TI - [Cognitive function in the aged. Little deterioration from moderate alcohol consumption]. PMID- 16038129 TI - [Early sequential therapy]. PMID- 16038130 TI - [Sinus thrombosis in heparin-induced thrombocytopenia type II]. PMID- 16038132 TI - [Ambulatory medicine trial. More damage than benefit?]. PMID- 16038131 TI - [Inflammatory lung and systemic disease]. PMID- 16038133 TI - [Schizophrenia. Psychoeducation as "travel leader in the land of improvement"]. PMID- 16038134 TI - [Adipokines--a future for therapeutic use?]. PMID- 16038135 TI - [ Preparation of folic acid supplements]. PMID- 16038136 TI - [Fast diagnosis with MRI]. PMID- 16038137 TI - [Medicine and the media]. PMID- 16038138 TI - [Two years of legal practice of euthanasia in Belgium: comparison with the Netherlands. First evaluation in a palliative care unit]. AB - The Belgian law relative to euthanasia prescribes that a physician performing an euthanasia has to complete a registration document and to send it within 4 days to the federal commission for control and evaluation of euthanasia. The 259 first documents are described in the report of the commission referred to Parliament on September 17, 2004. The present paper analyses this report and compares its most important data with those published in the Netherlands: apart from the total number of euthanasia's, much smaller in Belgium, and apart from aspects which are specifically related to the Belgian law, the statistical data are very similar in both countries. The difference in the number of registration documents written in French and in Flemish is analyzed and discussed. A first evaluation of the application of the law in a supportive and palliative care unit is reported. PMID- 16038139 TI - [Docetaxel as salvage treatment for non-small cell lung cancer: implementation study]. AB - Salvage treatment with docetaxel after failure of cisplatin-based chemotherapy in non-small cell lung cancer (NSCLC) has already been tested in the context of clinical prospective trials but only dealing with selected patients. The aim of the present study is to assess the generalizability of these results in an unselected population. We retrospectively analysed the data of all patients not included in clinical studies and treated at the Jules Bordet Institute between September 1996 and July 2003 by docetaxel after a previous chemotherapy containing platinum. Patients were separated in two groups: eligible or not studies according to the selection criteria of the published phase II and III prospective trials. There were 27 patients treated by docetaxel after relapse or failure of prior first-line platinum-based chemotherapy. Among them, 15 were ineligible according to the criteria of published phase II and III trials. The global response rate was 7.41%. All responders had a partial response to first line chemotherapy. Toxicity consisted mainly in grade III/IV neutropenia and was similar between the two group of patients. The median survival time was 25 weeks. In conclusion, our implementation study confirms the activity of docetaxel as salvage therapy after failure of first line cisplatin-based chemotherapy in an unselected population of patients with NSCLC, with a response rate similar to that obtained in propective trials. PMID- 16038140 TI - [The endocannabinoid system and the regulation of the metabolism]. AB - The endocannabinoid system modulates many physiological functions by acting on receptors CB1 and CB2. The endocannabinoids are produced only when and where they are needed. They act locally and are immediately metabolised after their action. Overactivation of the endocannabinoid system is observed in obesity, with stimulation of the appetite in the hypothalamus and fat accumulation in the adipocytes with increase of insulin resistance and decrease of adiponectin. Nicotine use overactivates also the endocannabinoid system. CB1 blockade by a specific inhibitor (rimonabant) decreases food intake and weight in animal studies and increases adiponectin and insulin sensitivity. Moreover, rimonabant decreases tobacco dependence. Clinical studies with rimonabant are encouraging. PMID- 16038141 TI - [Rimonabant (Acomplia), specific inhibitor of the endocannabinoid system]. AB - The endocannabinoid system plays a major role in the regulation of body energy by stimulation of the appetite in the hypothalamus and increase of fat accumulation in adipocytes. The blockade of the cannabinoid system (CB1) by the specific inhibitor (rimonabant) decreases food intake and adiposity in animals and in humans. Moreover rimonabant lowers tobacco addiction. Clinical studies (RIO LIPIDS and RIO-EUROPE) have recently confirmed that rimonabant combined with a hypocaloric diet over 1 year, promoted significant decrease of body weight, waist circumference and improvement of dyslipidemia. Rimonabant was well tolerated with mild and transient side effects. The future place of rimonabant in the strategy of obesity is still to be clarified. PMID- 16038142 TI - [Buccal ulcerations in an Ecuadorian patient]. AB - The goal of this text is to remind the place of histoplasmosis in the differential diagnosis of chronical buccal lesions, even if this pathology is extremely rare in our countries. The diagnosis is easy and fast because we can do biopsy in a place easily accessible. PMID- 16038143 TI - [Complications during lithium maintenance therapy]. AB - Through the case report of a bipolar patient treated with lithium for the past twenty years who presented a nephrogenic diabetes insipidus, a renal insufficiency, and a lithium intoxication. The different complications and their eventual treatment of lithium maintenance therapy are exposed and discussed. PMID- 16038144 TI - [Prescription in International Common Denomination. Affirmation of the pharmacist's competence]. PMID- 16038145 TI - [Prescription in International Common Denomination. A new challenge for general physicians?]. PMID- 16038146 TI - [Hippocrates, father of semiology and medical deontology]. AB - Elaborated 24 centuries ago, Hippocrate's and his school's works are considerable. They concern all aspects of medicine, surgery and even obstetrics, tackled by three approaches: clinical observation by constitution for each patient of what can be considered as the ancestor of the current medical file and by use of clinical signs still in application in modern semeiology; the Oath, basis of benevolence ethic, still taken nowadays but in an actualized form by future medical doctors; humoral theory which proposes a physiopathological concept of diseases and deduces a therapeutical approach still in use until the end of 19th century, by lack of sufficient knowledge about organs function and origin of diseases. It consists in an amalgam of archaic concepts, elaborated by presocratic philosophers about matter's structure transposed to body fluids and presented in a pseudocoherent form. Current concepts of diseases, especially role of pathogens render it obsolete. PMID- 16038147 TI - Health promotion: the next generations. PMID- 16038148 TI - Charters, declarations, world conferences: practical significance for health promotion practitioners 'on the ground'. PMID- 16038149 TI - Bridging the gap: integrating quality management into health promotion practice. PMID- 16038150 TI - Therapeutic injections in the context of Egyptian culture. PMID- 16038151 TI - HIV education in rural China. PMID- 16038152 TI - [Heath promotion today: proposals for the future]. PMID- 16038153 TI - [Collaborative effort between UIPES and WHO--Interview with Dr. Robert Beaglehole]. PMID- 16038154 TI - [Cough and malaise in young children due to allergic rhinoconjunctivitis]. AB - Four young children (a 6-year-old girl and three boys in the ages of 3.5, 4 and 10 years) with allergic rhinoconjunctivitis were referred with symptoms of cough and general discomfort. Examination revealed signs and symptoms of allergic rhinoconjunctivitis. Blood tests revealed that they were allergic to house dust mite. The symptoms decreased following suitable treatment with antihistamines or nasal corticosteroids. Allergic rhinoconjunctivitis is usually diagnosed in adolescents and adults with symptoms of sneezing, nasal congestion or rhinorrhea and itchy red eyes ('hay fever'). In view of the availability of effective treatment options, it is important to recognise this condition in children who present with cough and malaise. PMID- 16038155 TI - [The practice guideline 'Lower urinary-tract symptoms in middle-aged and elderly men' (second revision) from the Dutch College of General Practitioners; a response from the perspective of general practice]. AB - Recent research has shown that the simple pathophysiological concept which explains micturition complaints in elderly men as being caused by benign prostatic hyperplasia needs to be abandoned. It has been shown that there is no clear correlation between prostate volume and voiding problems and also that reductase blocker drugs which specifically decrease prostate volume do not lead to a clinically important reduction in symptoms. The internationally accepted new conceptual umbrella description of these micturition problems is 'lower urinary tract symptoms', a term that is more suited to research this area where a good understanding of the fundamental underlying mechanisms is lacking. The Dutch general practitioners' guideline for micturition problems in elderly men addresses this paradigm change in a thorough and easily understandable way. PMID- 16038156 TI - [The practice guideline 'Lower urinary-tract symptoms in middle-aged and elderly men' (second revision) from the Dutch College of General Practitioners; a response from the perspective of urology]. AB - New insights in the fields of terminology, diagnosis and treatment have been successfully implemented in the second revision of the practice guideline 'Lower urinary-tract symptoms in middle-aged and elderly men' from the Dutch College of General Practitioners. The result is a coherent, lucid and moreover practical guideline. It is important to stress that lower urinary-tract symptoms do not increase the patient's risk of prostate cancer. Screening for prostate cancer by the routine measurement of serum prostate-specific antigen is therefore not indicated. PMID- 16038157 TI - [Policy decisions from a societal perspective: local or national evaluation of health related quality of life not necessary]. AB - Research among a sample of the Dutch population has shown that different relative values are assigned to some of the five sub-domains of the 'Euro-Quality of life, five dimensions' (EQ-5D) questionnaire when compared to the current English values. These findings might be interpreted as proof that the resulting 'tariffs' should also differ across countries. Conversely, one could argue that the current Oxford tariffs are simply inadequate. Clearly, decisions for individual patients should be based on the individual's preferences. However, policy decisions taken from a societal perspective should be based on a global value system. It would seem that a single Western European tariff would be preferable to a whole range of national tariffs. PMID- 16038158 TI - [The possible role of microchimerism in the aetiology of autoimmune diseases]. AB - The aetiology of several autoimmune diseases has not yet been elucidated. Microchimerism, the persistence of small numbers ofallogeneic cells in an individual, has been mentioned recently in connection with the occurrence of autoimmune diseases such as systemic sclerosis and juvenile inflammatory idiopathic myopathy. These allogeneic cells can originate from mutual foeto maternal exchange of blood cells during pregnancy or from a donor after blood transfusion or (organ) transplantation. In some cases, a syndrome then develops that resembles the chronic graft-versus-host reaction after stem-cell transplantation, in which allogeneic cells react with autologous cells. Studies on microchimerism in patients with systemic sclerosis and juvenile inflammatory idiopathic myopathy, compared to controls, sometimes reveal a clearly increased prevalence of microchimerism in patients. However, microchimerism can also be found in healthy individuals. Direct proof of a causal relation between microchimerism and autoimmune diseases does not exist. Additional genetic or environmental factors may be partly responsible for a disturbed balance between tolerance and aggression. PMID- 16038159 TI - [The feasibility of the razor-blade shave technique in the diagnosis and treatment of benign and malignant skin lesions on the face]. AB - The razor-blade shave technique uses a slightly-curved classic razor blade to shave and remove the exophytic part of a skin lesion and part of the intradermal structure down to the stratum papillare. It can then be sent for pathology investigations. This treatment is curative for many benign skin conditions. The cosmetic result ofthe razor-blade technique is superior to that of surgical excision. The razor-blade technique is useful as a biopsy instrument in the diagnosis of skin rumours of unknown nature, especially in keratoacanthomas. The technique is very easy and cheap. However, if the skin lesion is a suspected malignant melanoma, traditional surgical excision should be performed. The shave technique is not suitable for malignant skin tumours. PMID- 16038160 TI - [Summary of the practice guideline 'Lower urinary-tract symptoms in middle-aged and elderly men' (second revision) from the Dutch College of General Practitioners]. AB - The 1997 practice guideline from the Dutch College of General Practitioners concerning lower urinary-tract symptoms (LUTS) in middle-aged and elderly men has been revised and some points have been adapted. The underlying cause of LUTS in middle-aged and elderly men is an improperly functioning voiding mechanism of the bladder associated with ageing. Symptoms are not simply due to prostate enlargement. In uncomplicated LUTS the patient's perception of the level of inconvenience is very important in considering and choosing therapeutic options. Percussion of the bladder after micturition is no longer universally advised. In general, invasive treatment is more effective in relieving symptoms than medical treatment, although invasive treatment causes more adverse effects. LUTS and prostate cancer are different entities, and LUTS is not a risk factor for prostate cancer. The issue of prostate cancer is discussed in this practice guideline in order to clear up popular misconceptions and to enhance the practical implementation of this guideline. PMID- 16038161 TI - [Diagnostic image (241). A woman with malaise, periodic fever and extreme fatigue]. AB - A 51-year-old white woman with extreme fatigue, fever episodes, weight loss, leucocytosis and increased erythrocyte sedimentation rate showed diffuse high uptake of 18F-fluorodeoxyglucose in the entire aortic wall, which is characteristic of giant cell large-vessel arteritis, such as Takayasu disease. PMID- 16038162 TI - [Measuring the quality of life in economic evaluations: the Dutch EQ-5D tariff]. AB - OBJECTIVE: To value EQ-5D health states by a general Dutch public. EQ-5D is a standardised questionnaire that is used to calculate quality-adjusted life-years for cost-utility analysis. DESIGN: Descriptive. METHOD: A sample of 309 Dutch adults from Rotterdam and surroundings was asked to value 17 EQ-5D health states using the time trade-off method. Regression analysis was applied to the valuations of these 17 health states. By means of the estimated regression coefficients, which together constitute the so-called Dutch tariff, valuations can be determined for all possible EQ-5D health states. These values reflect the relative desirability of health states on a scale where 1 refers to full health and 0 refers to death. Societal valuations are necessary in order to correct life years for the quality of life. RESULTS: Complete data were obtained from 298 persons. Theywere representative for the Dutch population as far as age, gender and subjective health were concerned, but had a somewhat higher educational level. The estimated Dutch EQ-5D tariff revealed that the respondents assigned the most weight to (preventing) pain and anxiety or depression, followed by mobility, self-care and the activities of daily living. The Dutch tariff differed from the UK ('Measurement and Valuation of Health') tariff, which is currently used in Dutch cost-utility analyses. Compared to UK respondents, Dutch respondents assigned more weight to anxiety and depression and less weight to the other dimensions. Conclusion. The valuation of health states by this representative Dutch study group differed from the valuation that is currently used in Dutch cost-utility analyses. PMID- 16038163 TI - [Evaluation of the rate of autopsy and rate of disparity between autopsy results and clinical cause of death in a surgical ward, with the emphasis on necrological review]. AB - OBJECTIVE: To determine the rate of autopsy and the rate of disparity between autopsy results and the clinically determined cause of death in a surgical ward. DESIGN: Descriptive. METHOD: A total of 12,000 patients were admitted to the surgical ward of the Red Cross Hospital, the Hague, the Netherlands, from January 1999 to December 2002. 305 (3%) died during their stay on the ward. By using our standard mortality registration system, it was possible to classify the causes of death, evaluate shortcomings in treatment, and determine the extent of agreement between pre- and post-mortem findings. RESULTS: Permission for an autopsy was obtained for 136 patients (45%). The autopsy rates in patients who died following abdominal aortic surgery, colonic surgery, peripheral artery bypass surgery, and hip surgery were 55%, 63%, 35% and 30%, respectively. In 37 patients (27%), the autopsy report revealed a disparity with the clinical cause of death. Patients who died after abdominal aortic surgery or colonic surgery had disparity rates of 33% and 21%, respectively. Patients who died after peripheral artery bypass surgery or hip surgery had disparity rates of 13% and 7%, respectively. CONCLUSION: The overall rate of autopsy was lower (45%) than in the period 1992 1998 (60%), but remained relatively high in patient groups who were previously found to have a high rate of disparity between pre- and post-mortem findings. Post-mortem examination remains an important tool that can be used to verify diagnosis and treatment and therefore assess the quality of care. PMID- 16038164 TI - [Liver abscesses as a complication of Crohn's disease]. AB - Liver abscesses were found in two women aged 23 and 34 years who suffered from Crohn's disease. The first patient was seen because of fever and thoracic pain and had been treated with infliximab. The second patient, who was pregnant, presented with abdominal pain that was thought to be due to an exacerbation of her inflammatory bowel disease. Ultrasonography and CT revealed that both patients had large liver abscesses. Both received antibiotic treatment, the first patient underwent drainage of the abscess, and the second underwent puncture twice, resulting in clinical improvements in both patients. In contrast to intra abdominal abscesses, liver abscesses are rarely seen in patients with Crohn's disease. The clinical presentation can be mistaken for an exacerbation of Crohn's disease, but the diagnosis can be made easily using ultrasonography or CT. Treatment consists of (ultrasound-guided) percutaneous drainage and administration of antibiotics. PMID- 16038165 TI - [Liver abscess following navel piercing]. AB - In an 18-year-old woman with abdominal complaints of unknown origin, ultrasonography and CT showed a large abnormal structure in the left half of the liver. Biopsy showed this to be an abscess. Because Staphylococcus aureus was cultured and the patient had suffered from a navel infection after navel piercing several months earlier, it was concluded that the infection had spread through the ligamentum teres hepatis to cause the liver abscess. The abscess was drained and the patient was treated successfully with antibiotics. PMID- 16038166 TI - [Eosinophilic oesophagitis in young men with food swallowing complaints]. PMID- 16038167 TI - [Pseudogout in 3 patients with presumed therapy-resistant rheumatoid arthritis]. PMID- 16038168 TI - [Lithium, a potentially dangerous drug]. PMID- 16038169 TI - Lessons learned in drug development from anesthesia. PMID- 16038170 TI - [Effectiveness of postoperative autologous blood transfusion after knee replacement surgery and influence of preoperative hemoglobin level]. AB - OBJECTIVE: To evaluate the effectiveness of postoperative autologous blood transfusion on reducing the need for allogenic transfusion during recovery from total knee arthroplasty until hospital discharge, and to determine whether effectiveness is related to preoperative hemoglobin level. MATERIAL AND METHODS: Retrospective study of patients undergoing surgery at Hospital Jerez de la Frontera, Spain, in 2003, assessing the association between postoperative autologous blood transfusion, preoperative hemoglobin, and allogenic transfusion requirements. RESULTS: A total of 107 patients were studied. Eighty-three received autologous blood transfusions after surgery and 15 (14.02%) required allogenic transfusion. The rate of allogenic transfusion was higher in association with hemoglobin levels exceeding 13 g x dL(-1) (P=0.003) and it was lower in patients who received autologous blood transfusions (P=0.046). In patients who received autologous transfusion, preoperative hemoglobin level and risk of allogenic transfusion were unrelated. When autologous transfusion was not given, allogenic transfusion risk was higher when hemoglobin concentration was less than 13 g x dL(-1) (P=0.0008). Autologous transfusion had a significant effect when hemoglobin level was less than 13 g x dL(-1) (P=0.002) but did not affect the rate of transfusion when hemoglobin was 13 g x dL(-1) or more. CONCLUSIONS: Autologous blood transfusion is effective for reducing the need for allogenic transfusion after knee replacement surgery, particulary when a patient's hemoglobin level is less than 13 g x dL(-1). PMID- 16038171 TI - [Evaluation of the OrthoPAT autologous transfusion system by experimental models simulating intra- and postoperative blood salvage]. AB - OBJECTIVE: To evaluate the efficacy of the OrthoPAT (Haemonetics) system for blood salvage and for removing chemical or cellular debris, by experimental models simulating intra- and postoperative conditions. MATERIAL AND METHODS: Blood samples (20%-25% packed red cells) were prepared for the intraoperative model (n=8) and the postoperative model (n=22). Surgical compresses were soaked in some samples (n=5). Other samples were supplemented with hemolyzed blood (n=7). From others cytokines were removed and blood activated with bacterial liposaccharides (n=10) was added. The samples were analyzed before and after processing; tests included detection of free plasma hemoglobin (FPH), potassium ions (K+), glutamic oxalic transaminase (GOT), lactate dehydrogenase (LDH), proteins, and cytokines. RESULTS: In the intraoperative model 2935 (SD 260) mL of blood was processed. The concentration of packed red cells was 63% and 80% of the red cells were recovered. In the postoperative model 652 (35) mL was processed, the packed red cell concentration was 67% and 81% of the red cells were recovered. Reductions were observed in the concentrations of white blood cells (72%), platelets (88%), GOT and LDH (75%), and proteins and K+ (>95%). Fifty percent of the red cells were recovered in the surgical compresses model. In the hemolysis model, the K+ and FPH concentrations were reduced more than 95%. In the cytokine model, up to 90% of the interleukin 1beta, interleukin 6, and tumor necrosis factor content was removed from the activated blood samples. CONCLUSIONS: These findings suggest that the OrthoPAT system washes blood and salvages content effectively, recovering 80% of red cells. Moreover, its processing capacity (800-1000 mL x h(-1)) seems adequate for blood replacement in orthopedic surgery. PMID- 16038173 TI - [Severe injury in the obstetric patient: considerations for anesthesiologist and critical care specialist]. AB - In spite of the high incidence of moderately severe or severe injury in obstetric patients, the anesthesiology literature contains few references to their diagnosis and treatment. We review the epidemiology, etiology, treatment, and prognosis of severe or moderately severe injuries in obstetric patients. PMID- 16038172 TI - [Comparison of gastric aspirate volume after intubation in patients with and without symptoms of gastroesophageal reflux disease]. AB - OBJECTIVES: To compare the postintubation gastric aspirate volume of patients with and without symptoms of gastroesophageal reflux disease (GERD). METHODS: Prospective randomized study of 331 physical status ASA I-II adults scheduled for surgery. Patients with GERD symptoms (heartburn and regurgitation) were assigned to group A (n=83); asymptomatic patients (n=248) were assigned to groups B (n=85), C (n=70), and D (n=93). Group A was subdivided: group A1 received outpatients treatment for GERD and group A2 did not. Groups A, B, and C received prophylaxis with omeprazole 40 mg and metoclopramide 10 mg, respectively. Group D received no prophylaxis. Groups A and B patients received 1 mg x kg(-1) of succinylcholine, and groups C and D received 0.2 mg x kg(-1) of cisatracurium. After intubation, gastric content was aspirated and measured. The results were compared with Pearson's chi2 and Student t tests, analysis of variance, and Kruskal-Wallis and Mann-Whitney U tests. RESULTS: The gastric aspirate volumes in each group were as follows: 36.6 (SD 5) mL in group A; 9.98 (4.9) mL in group B; 10.8 (5.5) mL in group C; 15.62 (6.3) mL in group D; 32.7 (5.1) mL in group A1; and 39.08 (3.6) mL in group A2. Volumes were significantly greater in group A than in groups B, C, and D (P<0.0001), and in subgroup A2 than in subgroup A1 (P<0.001). The percentages of patients from whom a volume greater than 25 mL was aspirated in each group were as follows: 98.9% of group A, 1.2% of group B, 2.8% of group C, and 8.6% of group D (P<0.001). CONCLUSION: Patients with chronic heartburn and regurgitation have greater postintubation gastric aspirate volumes than do asymptomatic patients, attributable to the gastroesophageal motility dysfunction characteristic of GERD that can favor aspiration. GERD questions should be included in preanesthetic medical history taking, and symptomatic patients should be prescribed outpatient proton pump inhibitor therapy. PMID- 16038174 TI - [The AGREE appraisal tool for assessing guidelines for postoperative pain management]. PMID- 16038175 TI - [Dual atrioventricular nodal conduction and arrhythmia with severe hemodynamic alterations during liver retransplantation]. AB - We report the case of a man who developed tachycardia caused by atrioventricular reentry related to dual nodal conduction during liver retransplantation. The hemodynamic alterations were severe. Arrhythmia and altered cardiac conduction are potential complications of liver transplantation. The development of tachyarrhythmias--atrial fibrillation as well as episodes of supraventricular and ventricular tachycardia and bradycardia--have been described. Such arrhythmias tend to occur particularly during reperfusion of the graft. Risk factors implicated are the severe ion imbalances, acid-base imbalance, and hypothermia that accompany the reperfusion of a new organ. A review of the possible pathogenic and etiological mechanisms that lead to arrhythmia in patients with end-stage liver disease is provided. PMID- 16038176 TI - [Hafnia alvei in a rare case of severe pneumonia in a postanesthesia recovery unit]. AB - An 85-year-old man with a history of moderate chronic obstructive pulmonary disease in treatment with anticholinergic drugs was admitted to the recovery unit for postoperative monitoring after right inguinal herniorrhaphy surgery and removal of a strangulated intestinal section. On the third day after surgery the patient developed radiographic signs consistent with pneumonia and required mechanical ventilation. Two blood cultures and a respiratory secretion sample grew a microorganism identified as Hafnia alvei. H. alvei is a gram-negative bacillus that colonizes the digestive tract of humans and animals and in immunodepressed patients it can colonize the mouth and pharynx. Isolation of H. alvei is described in the literature on pediatric patients and those with a history of immune deficiency or chronic disease. Infection has a severe impact on general health. We report a rare and interesting case of pneumonia and bacteremia from H. alvei infection acquired by an immunocompetent patient soon after arrival in the postoperative recovery unit. The patient died of the infection. PMID- 16038177 TI - [Nasotracheal fiberoptic intubation under remifentanil for sedation and analgesia in a boy with a difficult airway due to giant gingival hypertrophy]. AB - A 9-year-old boy diagnosed with gingival hypertrophy and with a history of West syndrome and associated low platelet levels underwent gingival reduction surgery. Because difficult intubation was foreseen, the fiberoptic tube was inserted through the nose with the patient breathing spontaneously under remifentanil for sedation and analgesia. The procedure was carried out under balanced general anesthesia and with standard monitoring. At the end of gingivectomy, a tracheostomy was performed and the patient was transferred to the pediatric intensive care unit for postoperative observation. PMID- 16038178 TI - [Intracoronary air embolism detected during intraoperative transesophageal echocardiography]. AB - A 39-year-old hypertensive man with severe aortic stenosis underwent aortic valve replacement monitored by intraoperative transesophageal echocardiography. Upon weaning the patient off extracorporeal circulation, hemodynamics became severely compromised, with hypotension, tachycardia, and elevated precordial electrocardiographic tracings. The echocardiographic images were instrumental during the episode to demonstrate that the anterior wall presented hypokinesis consistent with ischemia in the region but that there were also images of hyperrefringence highly suggestive of intracoronary air embolism. Intraoperative transesophageal echocardiography allowed us to diagnose the real cause of the ischemic event and rule out an atheromatous plaque as the source. Perfusion pressure was increased to treat the air embolism. The echocardiographic image demonstrated success, specifically restoration of left ventricular regional contractility. This experience revealed the usefulness of transesophageal echocardiography in intraoperative monitoring to diagnose ischemia, assess the cause, and guide treatment. PMID- 16038179 TI - [Subarachnoid anesthesia in a patient with alkaptonuric ochronosis]. PMID- 16038181 TI - [Epidural blood patch in a Jehovah's witness]. PMID- 16038180 TI - [Suspicion of pulmonary thromboembolism in a pregnant woman: differential diagnosis]. PMID- 16038182 TI - [Accidental catherization of the inferior vena cava with a Swan-Ganz catheter: radiologic monitoring]. PMID- 16038183 TI - [Perineal pruritus after intravenous injection of dexamethasone for postoperative prophylaxis of nausea and vomiting]. PMID- 16038184 TI - [When prophylaxis fails: Is treatment of postoperative nausea and vomiting with same-class antiemetics useful?]. PMID- 16038185 TI - Labor analgesia and idiopathic thrombocytopenic purpura. PMID- 16038186 TI - [Why use a posterior approach to the lumbar plexus block in a patient with hemostatic abnormalities?]. PMID- 16038187 TI - [Venous catheter migration to the pulmonary artery]. PMID- 16038188 TI - Sensitive and specific quantification of the anticancer agent ZD1839 (Gefitinib) in plasma by on-column focusing capillary liquid chromatography-tandem mass spectrometry. AB - The development of an on-column focusing gradient capillary LC method coupled to tandem mass spectrometry (quadrupole-linear ion trap) for the quantitative determination of the anticancer agent ZD1839 (Gefitinib, Iressa) in blood plasma is described. Plasma samples (0.2 ml) were extracted with methyl tert-butyl ether. The analytes of interest, ZD1839 and the internal standard [(2)H8]ZD1839 (ZD1839-d8) were eluted on a 50 mm x 1 mm, 5 microm particle size, capillary ODS Hypersil column using an aqueous ammonium acetate gradient at 40 microl/min. Mass spectrometric detection was performed by a Q-Trap tandem mass spectrometer with electrospray positive ionisation, and monitored in the multiple reaction monitoring transitions 447 >128 and 455 >136, respectively. The limit of quantification of ZD18395 was 0.1 ng/ml. The method proved to be robust, allowing quantification of ZD1839 with sufficient precision, accuracy and sensitivity. PMID- 16038189 TI - Identification and quantification of tamoxifen and four metabolites in serum by liquid chromatography-tandem mass spectrometry. AB - We have developed a method for the determination of tamoxifen (tam) and its metabolites 4-hydroxytamoxifen (4OHtam), N-demethyltamoxifen (NDtam), N dedimethyltamoxifen (NDDtam), tamoxifen-N-oxide (tamNox), and 4-hydroxy-N demethyltamoxifen (4OHNDtam) in 50 microl human serum. Serum proteins were precipitated with acetonitrile. Deuterated-tamoxifen (D5 tam) was added as internal standard. Sample supernatant was injected into an on-line reversed-phase extraction column coupled with a C18 analytical column and analytes were detected by tandem mass spectrometry. The lower limits of quantification were 0.25 ng/mL for 4OHtam, NDtam and tam, 1.0 ng/mL for NDDtam and tamNox. Ranges of within- and between-day variation were 2.9-15.4% and 4.4-12.9%, respectively. PMID- 16038190 TI - Quantitative analysis of delta9-tetrahydrocannabinol in preserved oral fluid by liquid chromatography-tandem mass spectrometry. AB - A rapid and sensitive method for the analysis of delta9-tetrahydrocannabinol (THC) in preserved oral fluid was developed and fully validated. Oral fluid was collected with the Intercept, a Food and Drug Administration (FDA) approved sampling device that is used on a large scale in the U.S. for workplace drug testing. The method comprised a simple liquid-liquid extraction with hexane, followed by liquid chromatography-tandem mass spectrometry (LC-MS-MS) analysis. Chromatographic separation was achieved using a XTerra MS C18 column, eluted isocratically with 1 mM ammonium formate-methanol (10:90, v/v). Selectivity of the method was achieved by a combination of retention time, and two precursor product ion transitions. The use of the liquid-liquid extraction was demonstrated to be highly effective and led to significant decreases in the interferences present in the matrix. Validation of the method was performed using both 100 and 500 MicroL of oral fluid. The method was linear over the range investigated (0.5 100 ng/mL and 0. 1-10 ng/mL when 100 and 500 microL, respectively, of oral fluid were used) with an excellent intra-assay and inter-assay precision (relative standard deviations, RSD <6%) for quality control samples spiked at a concentration of 2.5 and 25 ng/mL and 0.5 and 2.5 ng/mL, respectively. Limits of quantification were 0.5 and 0.1 ng/mL when using 100 and 500 microL, respectively. In contrast to existing GC-MS methods, no extensive sample clean-up and time-consuming derivatisation steps were needed. The method was subsequently applied to Intercept samples collected at the roadside and collected during a controlled study with cannabis. PMID- 16038191 TI - Bacterial response to eukaryotic cells. Analysis of differentially expressed proteins using nano liquid chromatography-electrospray ionization tandem mass spectrometry. AB - Host-bacteria interactions have mostly been investigated with regard to the host response or to activities of pathogenic bacteria. In contrast, we aim to identify reactions of non-pathogenic bacteria that result from their contact with host cells of the gastrointestinal tract. In a proteomic approach, the response of non pathogenic human Escherichia coli bacteria on gut epithelial cells (rat IEC-6) was investigated in an in vitro co-culture model. For this purpose, a sensitive analytical procedure was developed based on the identification of two-dimensional polyacrylamide gel electrophoresis separated proteins by online nanoLC electrospray ionization MS/MS using a quadrupole time-of-flight tandem mass spectrometer for accurate mass determination. We demonstrate here the efficiency of this technique by the identification of a total of 43 differentially expressed proteins, out of which 25 were up-regulated and 18 were down-regulated. They represent a wide range of molecular weight and different metabolic and physiological functions. PMID- 16038192 TI - On-line identification of secondary metabolites in freshwater microalgae and cyanobacteria by combined liquid chromatography-photodiode array detection-mass spectrometric techniques. AB - The analysis and identification of a wide range of secondary metabolites biosynthesized by different algal taxa and cyanobacteria has been performed through a selective and sensitive methodology, mainly based on reversed-phase HPLC coupled both to UV photodiode array detection and to atmospheric pressure mass spectrometric techniques (HPLC-DAD-APIMS). Results are reported here with special attention to the analyses carried out both on the natural phytoplankton (mixed populations) of Lake Tovel (Northern Italy, Brenta Dolomites) and on enclosure-produced biomass of the dinoflagellate Glenodinium sanguineum Marchesoni (1941). This analytical procedure might represent a powerful tool for the fast screening of the taxonomic composition (broad groups, e.g. divisions) of natural mixed populations of phytoplankton, by providing a reliable distribution of accessory pigments extracted from microalgae, such as carotenoids and chlorophyll derivatives. Furthermore, we showed that in the same chromatographic analysis other classes of natural products, such as galactolipids, alkaloids, sterols and mycosporine-like amino acids, can be detected by using combined optical and mass spectrometric techniques. These metabolites represent distinctive biochemical signatures, sometimes even at the species level. PMID- 16038193 TI - Determination of biogenic amines in fresh and processed meat by suppressed ion chromatography-mass spectrometry using a cation-exchange column. AB - A new method for simultaneous determination of underivatized biogenic amines based on the separation by cation-exchange chromatography and suppressed conductivity coupled with mass spectrometry detection has been developed. The method has been applied to the analysis of cadaverine, putrescine, histamine, agmatine, phenethylamine and spermidine in processed meat products. The amines were extracted from muscle tissue with methanesulfonic acid without any additional derivative step or sample clean-up. Biogenic amines were separated by the IonPac CS17 column, a cation-exchange column used with gradient elution, and detection was done by suppressed conductivity and mass spectrometry. Tyramine was simultaneously analysed by using a spectrophotometer (275 nm) before the suppressed conductivity detection. Linearity of response was obtained in the range 0.25-25 microg mL(-1). The detection limits ranged from 23 microg L(-1) for putrescine to 155 microg L(-1) for spermidine (suppressed conductivity) and from 9 microg L(-1) for agmatine to 34 microg L(-1) for spermidine (MS). Average recoveries from meat samples ranged from 85 to 97% and coefficients of variation ranged from 4.5 to 9.7%. The analysis of biogenic amines in fresh and processed meats (dry-cured, cooked and fermented products) can be used as a quality marker of raw material and for studying the relationship between their changes and the fermentation process involved in dry sausage ripening. PMID- 16038194 TI - On-line identification of sugarcane (Saccharum officinarum L.) methoxyflavones by liquid chromatography-UV detection using post-column derivatization and liquid chromatography-mass spectrometry. AB - Sugarcane (Saccharum officinarum L., Gramineae) bagasse and leaves were investigated for their flavonoid content and transgenic sugarcane ("Bowman-Birk" and "Kunitz") was compared with non-modified ("control") plants. Analyses were carried out by high-performance liquid chromatography coupled to diode array UV detection (LC/UV), also using post-column addition of shift reagents, and tandem MS (atmospheric pressure chemical ionization-MS/MS and collision-induced dissociation-MS). On-line UV and MS data demonstrated the presence of methoxyflavone glycosides and aglycones in a total of seven compounds. Three naturally occurring flavones glycosides and two unusual erythro- and threo diastereoisomeric flavolignan 7-O-glucosides were identified together with their aglycones. PMID- 16038195 TI - Quantitative determination and structural characterization of isoflavones in nutrition supplements by liquid chromatography-mass spectrometry. AB - In this paper, an accurate and route method was developed to quantitative determine daidzein, genistein, glycitein, daidzin, glycitin, 6"-O-acetyldaidzin, 6"-O-acetylglycitin and 6"-O-acetylgenistin contents in selected high and low isoflavones in nutrition supplements by on line liquid chromatography-atmospheric pressure chemical ionisation mass spectrometry (LC-APCI-MS). Improved extraction and hydrolysis methods of the isoflavones from three nutrition supplements were also studied and a rapid extraction method was developed. Comparison of different MS2 and MS3 spectra of isoflavones and some unknown compounds were also explored and proposed pathway fragments of nine isoflavones were first systematically suggested. PMID- 16038196 TI - Discovering metabolites of post-harvest fungicides in citrus with liquid chromatography/time-of-flight mass spectrometry and ion trap tandem mass spectrometry. AB - In this study, we benefit from the combination of liquid chromatography (LC)/time of-flight (TOF) MS accurate mass measurements to generate elemental compositions of ions and LC/ion trap multiple MS (MSn) providing complementary structural information, which is useful for the elucidation of unknown organic compounds at trace levels in complex food extracts. We have applied this approach to investigate different citrus fruits extracts, and we have identified two post harvest fungicides (imazalil and prochloraz), the main degradation product of imazalil ([M + H]+, m/z 257) and a non-previously reported prochloraz degradation product ([M + H]+, m/z 282). The database-mediated identification of the parent compounds was based on the generated elemental composition obtained from accurate mass measurements and additional qualitative information from the high resolution chlorine isotopic clusters of both the protonated molecules (imazalil, [M + H]+ 297.0556, <0.1 ppm error, 2-Cl; prochloraz, [M + H]+ 376.0381, 1.9 ppm error, 3 Cl) and their characteristic fragments ions (imazalil: m/z 255 and 159; prochloraz: m/z 308 and 266). The correlation between the structural information provided by ion trap MS/MS fragmentation pathways of the parent species and the TOF accurate mass elemental composition data of the degradation products were the key to elucidate the structures of the degradation products of both post-harvest fungicides. Finally, where standards were not available (prochloraz), further confirmation was obtained by synthesizing the proposed degradation product by acid hydrolysis of the parent standard and confirmation by LC/TOF-MS. PMID- 16038197 TI - Multi-residue pesticide analysis in fruits and vegetables by liquid chromatography-time-of-flight mass spectrometry. AB - In this work, a new multi-residue methodology using liquid chromatography-time-of flight mass spectrometry (LC-TOF-MS) for the quantitative (routine) analysis of 15 pesticide residues has been developed. The analytical performance of the method was evaluated for different types of fruit and vegetables: pepper, broccoli, tomato, orange, lemon, apple and melon. The accurate mass measurements were compared in different matrices at significantly different concentration levels (from 0.01 to 0.5 mg/kg) obtaining accuracy errors lower than 2 ppm, which is well within the accepted limits for elemental confirmation. Linearity of response over two orders of magnitude was demonstrated (r > 0.99). Matrix effects resulting in suppression or enhancement of the response were frequently observed, most notably in broccoli and citrus. Instrumental limits of detection (LOD) were between 0.0005 and 0.03 mg/kg depending on the commodity and pesticide studied, all being within European Union regulations for food monitoring program. Finally, the methodology was applied to the analysis of two samples from an inter laboratory exercise. The high degree of confirmation for target pesticides by accurate mass measurements demonstrated the applicability of the method in routine analysis. This study is a valuable indicator of the potential of LC-TOF MS for quantitative multi-residue analysis of pesticides in vegetables and fruits. PMID- 16038198 TI - Strategies to avoid the mis-identification of anatoxin-a using mass spectrometry in the forensic investigation of acute neurotoxic poisoning. AB - Anatoxin-a (AN) is a potent neurotoxin, produced by a number of cyanobacterial species, and consumption of freshwater contaminated with this toxin has led to animal deaths. Forensic investigations of suspected AN poisonings are frequently hampered by difficulties in detecting this toxin in biological matrices due to its rapid decay. In addition, detection of AN using single quadrupole mass spectrometry (MS) is suspect due to the presence of the amino acid, phenylalanine (Phe), since these compounds are isobaric and elute similarly in reversed phase liquid chromatography (LC). Approaches to prevent the misidentification of AN that have been explored in these studies included: (a) fluorimetric LC following derivatisation using 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F); (b) methylation using diazomethane prior to LC-MS determination; (c) multiple tandem MS using a quadrupole ion-trap (LC-MS3); and (d) hybrid quadrupole time-of-flight (QqTOF). Interference from Phe was not observed in any of procedures, (a)-(c), and the high mass accuracy obtained in method (d), readily distinguished between AN (165.11536) and Phe (165.07898). LC-MSn was also employed to study the fragmentation pathway of Phe and multi-stage MS spectra provided characteristic fragmentation information that clearly distinguished between AN and Phe. The difficulties associated with the over reliance on low resolution MS without MS/MS data in forensic toxicology are discussed. PMID- 16038199 TI - Probing new approaches using atmospheric pressure photo ionization for the analysis of brominated flame retardants and their related degradation products by liquid chromatography-mass spectrometry. AB - Atmospheric pressure photo ionisation has been evaluated for the analysis of brominated flame retardants and their related degradation products by LC-MS. Degradation mixtures obtained from the photochemical degradation of tetrabromobisphenol A and decabromodiphenylether were used as model systems for the assessment of the developed methodology. Negative ion mode gave best results for TBBPA and its degradation compounds. [M - H]- ions were formed without the need of using a doping agent. MS and MS/MS experiments allowed the structural identification of new TBBPA "polymeric" degradation compounds formed by attachment of TBBPA moieties and/or their respective cleavage products. In the case of polybromodiphenylethers, the positive mode provided M*+ ions and gave better results for congeners ranging from mono- to pentabromodiphenylethers whereas for higher bromination degrees, the negative ion mode (providing [M - Br + O]- ions) was best suited. Under both positive and negative ionisation modes, the use of toluene as doping agent gave better results. Liquid chromatography mass spectrometry by means of atmospheric pressure photo-ionisation was applied to the analysis of aromatic brominated flame retardants and their degradation products. This methodology proved to be particularly useful, for the characterisation and structural identification of some compounds which are not amenable to GC-MS, especially in the case of apolar "polymeric" degradation products of tetrabromobisphenol A investigated in this work. PMID- 16038200 TI - Stability of fluorinated surfactants in advanced oxidation processes--A follow up of degradation products using flow injection-mass spectrometry, liquid chromatography-mass spectrometry and liquid chromatography-multiple stage mass spectrometry. AB - The advanced oxidation process (AOP) reagents ozone (O3), O3/UV, O3/H2O2, and H2O2/Fe2+ (Fenton's reagent) were applied to the anionic and the non-ionic fluorinated surfactants perfluorooctanesulfonate (PFOS) and N-ethyl-N (perfluoroalkyl)-sulfonyl-glycinic acid (HFOSA-glycinic acid) or N-ethyl-N perfluoroalkyl sulfonylamido-2-ethanol polyethoxylates (NEtFASE-PEG), their methyl ethers (NEtFASE-PEG methyl ether) and partly fluorinated alkyl-ethoxylates (FAEO) dissolved in ultrapure water. To monitor the efficiencies of destruction samples were taken during the treatment period of 120 min. After sample concentration by C18-solid phase extraction (SPE) and desorption MS, coupled with atmospheric pressure chemical ionisation (APCI) or electrospray interface (ESI) was applied for detection. No elimination of PFOS was observed while HFOSA glycinic acid and AOP treated non-ionic surfactants were eliminated by oxidation. Degradation products could be detected and identified. So PFOS was observed during HFOSA-glycinic acid oxidation. Polyethylene glycols (PEG) and PEG methyl ethers were generated from non-ionic fluorinated surfactants beside their oxidation products--aldehydes and acids--all identified by tandem (MS-MS) or multiple stage mass spectrometry (MSn). AOP treatment of FAEO blend resulted in a mixture of partly fluorinated alcohols, separated and identified using GC-MS. PMID- 16038201 TI - Autoanalyzer for continuous fractionation and quantitation of the polyphenols content in wines. AB - A simple continuous flow autoanalyzer for the on-line fractionation of the polyphenols content in wines is proposed. The target compounds are isolated from the matrix by solid-phase extraction on an RP-C18 sorbent column, using selective solvents for the sequential elution of each polyphenol family. Moreover, evaporative light scattering detection (ELSD) is used for the first time for the on-line monitorization of the three polyphenol fractions present in the wine samples. Thus, a single sample injection is required to determine the global concentration of the three selected polyphenol fractions and the whole analysis is completed within a few minutes. Three calibration graphs were constructed for quantitative analysis of the global compounds concentration in every fraction and covered the range 5-300 mg l(-1) (expressed as gallic acid). Average repeatability, expressed as relative standard deviation, was 4%. The proposed autonalyzer was applied to the analysis of a variety of commercial wine samples. The results obtained were compared with those provided by the Folin-Ciocalteau method, being similar in all instances. PMID- 16038202 TI - Investigation on conformational order and mobility of diamondbond-C18 and C18 alkyl modified silica gels by Fourier transform infrared and solid-state NMR spectroscopy. AB - The effect of surface coverage and solid supports on the conformational order of alkyl chains of commercially available carbon clad zirconia based supports and synthesised C18-alkyl modified silica based supports are probed in the dry state for the first time using variable temperature Fourier transform infrared (FT-IR) and solid-state 13C NMR spectroscopy. From FT-IR spectroscopy, the conformational order of alkyl chains tethered to the substrates is examined by the analysis of CH2 symmetric and anti-symmetric stretching bands. Through solid-state 13C NMR spectroscopy, the order is inferred from the relative intensity of the main methylene carbon resonance assigned to trans and trans-gauche conformations. It is found that molecules tethered to the graphite layer experience a strongly diamagnetic component of the highly anisotropic magnetic susceptibility of the graphite lattice, which reflects upfield shift in the 13C NMR spectra of commercially available octadecyl-modified carbon clad zirconia based column materials. The present results prove that temperature, surface coverage and solid supports have an influence on the conformational order and mobility of alkyl chains tethered to the carbon clad inorganic metal oxides. PMID- 16038203 TI - Coupling of size-exclusion chromatography to a continuous assay for subtilisin using a fluorescence resonance energy transfer peptide substrate: testing of two standard inhibitors. AB - Liquid chromatography (LC) was coupled on-line to a homogeneous continuous-flow protease assay using fluorescence resonance energy transfer (FRET) as a readout for the screening of inhibitors of an enzyme (e.g., Subtilisin Carlsberg). The inhibitors aprotinin (a protein of approximately 6500 g/mol) and 4-(2 aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF, 240 g/mol) were mixed with other, non-active compounds and separated on a size-exclusion chromatography column. After the separation, the analytes were eluted to the postcolumn reactor unit where the enzyme solution and subsequently the FRET peptide substrate were added; by measuring the fluorescence intensity the degree of inhibition was monitored on-line. As expected, only the two inhibitors caused a change in the FRET response. Detection limits for aprotinin were 5.8 microM in the flow injection analysis (FIA) mode and 12 microM in the on-line LC mode. System validation was performed by determining IC50 values for aprotinin for the FIA mode (19 microM) and the on-line mode (22 microM). These IC50 values were in line with the value determined in batch experiments (25 microM). With this system, chemical information (i.e., chromatographic retention time) and biological information (i.e., enzyme inhibition) can be combined to characterize mixtures. PMID- 16038204 TI - Residue determination of glyphosate, glufosinate and aminomethylphosphonic acid in water and soil samples by liquid chromatography coupled to electrospray tandem mass spectrometry. AB - This paper describes a method for the sensitive and selective determination of glyphosate, glufosinate and aminomethylphosphonic acid (AMPA) residues in water and soil samples. The method involves a derivatization step with 9 fluorenylmethylchloroformate (FMOC) in borate buffer and detection based on liquid chromatography coupled to electrospray tandem mass spectrometry (LC-ESI MS/MS). In the case of water samples a volume of 10 mL was derivatized and then 4.3 mL of the derivatized mixture was directly injected in an on-line solid phase extraction (SPE)-LC-MS/MS system using an OASIS HLB cartridge column and a Discovery chromatographic column. Soil samples were firstly extracted with potassium hydroxide. After that, the aqueous extract was 10-fold diluted with water and 2 mL were derivatized. Then, 50 microL of the derivatized 10-fold diluted extract were injected into the LC-MS/MS system without pre-concentration into the SPE cartridge. The method has been validated in both ground and surface water by recovery studies with samples spiked at 50 and 500 ng/L, and also in soil samples, spiked at 0.05 and 0.5 mg/kg. In water samples, the mean recovery values ranged from 89 to 106% for glyphosate (RSD <9%), from 97 to 116% for AMPA (RSD < 10%), and from 72 to 88% in the case of glufosinate (RSD < 12%). Regarding soil samples, the mean recovery values ranged from 90 to 92% for glyphosate (RSD <7%), from 88 to 89% for AMPA (RSD <5%) and from 83 to 86% for glufosinate (RSD <6%). Limits of quantification for all the three compounds were 50 ng/L and 0.05 mg/kg in water and soil, respectively, with limits of detection as low as 5 ng/L, in water, and 5 microg/kg, in soil. The use of labelled glyphosate as internal standard allowed improving the recovery and precision for glyphosate and AMPA, while it was not efficient for glufosinate, that was quantified by external standards calibration. The method developed has been applied to the determination of these compounds in real water and soil samples from different areas. All the detections were confirmed by acquiring two transitions for each compound. PMID- 16038205 TI - Mathematical analysis of affinity membrane chromatography. AB - A mathematical model including convection, diffusion and Freundlich adsorption is developed. To examine the validity of the model, the affinity membranes were prepared by coating chitosan on the nylon membranes, a ligand of poly-L-lysine was bound to the chitoan-coating membranes, and the adsorption behavior of bilirubin through the stacked affinity membranes was investigated. The agreements between the theoretical and experimental results are exceptional. Using our new model, we show that: (1) As Pe increases, the breakthrough curves become sharper. For Pe greater than 30, the effect of axial diffusion is insignificant; (2) As m increases, the time of total saturation is delayed and the loading capacity at the point of breakthrough is increased; (3) As n decreases, the time of total saturation is delayed and the loading capacity at the point of breakthrough is increased; (4) As r increases, both the time of total saturation and the loading capacity at the point of breakthrough are increased; (5) adsorption rate influences the time of total saturation strongly but contributes little to the loading capacity. PMID- 16038206 TI - Characterization of immobilized artificial membrane (IAM) and XTerra columns by means of chromatographic models. AB - Immobilized artificial membranes (IAMs) prepared from phosphatidylcholine analogs are used as stationary phases in liquid chromatography systems to model drug partitioning between an aqueous phase (mobile phase) and a cell membrane (IAM column). Two different chromatographic models, which describe retention as a function of solute and column-mobile phase properties, have been applied to characterization of an IAM and two reversed phase C18 columns (Waters XTerra MSC18 and XTerra RP18) with acetonitrile-water mobile phases. The comparison of the results shows that the phosphatidylcholine group makes IAM column more polar than both XTerra columns, specially in terms of hydrogen-bond acceptor ability. XTerra RP18 is slightly more polar than XTerra MSC18 because of the presence of the embedded carbamate polar group. PMID- 16038207 TI - Development and validation of a hydrophilic interaction chromatography-mass spectrometry assay for taurine and methionine in matrices rich in carbohydrates. AB - A new procedure based on hydrophilic interaction chromatography coupled to tandem mass spectrometry (ionisation process by pneumatically assisted electrospray in negative ion mode), is developed and validated for the simultaneous determination of underivatised taurine and methionine in beverages rich in carbohydrates such as energy drinks. No initial clean-up procedure and no sample derivatisation are required. Satisfactory analysis was obtained on an Astec apHera NH2 (150 mm x 4.6 mm; 5 microm) column with methanol-water (60/40) as mobile phase. The method was validated in terms of specificity, detection limits, linearity, accuracy, precision and stability, using threonine as internal standard. The potential effects of matrix and endogenous amino acid content were also examined. The limits of detection in the beverage varied from 20 microg L(-1) for taurine to 50 micro L(-1) for methionine. PMID- 16038208 TI - Alpha-ribazole, a fluorescent marker for the liquid chromatographic determination of vitamin B12 in foodstuffs. AB - A method to determine the contents of free vitamin B12 in various foods by reversed phase liquid chromatography-fluorimetry is reported. It includes a purification of the samples by passage through an immunoaffinity column and a pre column conversion of vitamin B12 into the fluorescent alpha-ribazole (successive treatments of the extract with 2.5 M sodium hydroxide (at 100 degrees C for 15 min) and alkaline phosphatase (7.5 U) at 37 degrees C and pH 8 for 16 h). An enzymatic hydrolysis prior to the purification step (pepsin at 37 degrees C and pH 4 for 3 h) made it possible to release the vitamin B12 bound to proteins and thus to obtain the total vitamin B12 contents of these foodstuffs. The method proposed for the determination of free and bound vitamin B12 gives a good recovery rate (95-100%) and a satisfactory repeatability (R.S.D.r between 1.0 and 5.4%). Owing to its low quantification limit (3 ng g(-1)) and the good resolution of the alpha-ribazole peak, it could most probably be applied to the determination of this vitamin in any foodstuff. PMID- 16038209 TI - Hydrophilic interaction liquid chromatography--mass spectrometry for the analysis of paralytic shellfish poisoning (PSP) toxins. AB - Hydrophilic interaction liquid chromatography (HILIC) was examined for the separation of paralytic shellfish poisoning (PSP) toxins using the stationary phase TSK-gel Amide-80. The parameters tested included type of organic modifier and percentage in the mobile phase, buffer concentration, pH, flow rate and column temperature. Using mass spectrometric (MS) detection, the HILIC column allowed the determination of all the major PSP toxins in one 30 min analysis with a high degree of selectivity and sensitivity. The high percentage of organic modifier in the mobile phase and the omission of ion pairing reagents, both favored in HILIC, provided limits of detection (LOD) in the range 50-100 nM in selected ion monitoring (SIM) mode on a single quadrupole LC-MS system. LOD in selected reaction monitoring (SRM) mode on a sensitive triple quadrupole system were as low as 5-30 nM. Excellent linearity of response was observed. PMID- 16038210 TI - Evaluation of diphenylamine derivatives in apple peel using gradient reversed phase liquid chromatography with ultraviolet-visible absorption and atmospheric pressure chemical ionization mass selective detection. AB - A method was developed for extracting, identifying, and quantifying diphenylamine (DPA) derivatives in the peel of DPA-treated apples using gradient reversed-phase liquid chromatography with ultraviolet-visible absorption and atmospheric pressure chemical ionization detection (LC-UV-vis-APCI-MS). Compounds routinely analyzed using this method included hydroxylated, nitrosated, nitrated, and methoxylated diphenylamine derivatives. Analysis of peel treated with 0-8 g L(-1) DPA showed that peel DPA content was a limiting factor in derivative production and that recovery of most compounds over this range was linear. PMID- 16038211 TI - Validation of a screening method based on liquid chromatography coupled to high resolution mass spectrometry for analysis of perfluoroalkylated substances in biota. AB - A screening method for analysis of perfluoroalkylated substances (PFAS) in biota samples has been developed and validated using liver samples from polar cod (Boreogadus saida) and glaucous gull (Larus hyperboreus). The method was based on extraction of target compounds from homogenised samples into the solvent mixture used as mobile phase in high-performance liquid chromatography (HPLC), i.e. methanol/water (50:50; 2 mM ammonium acetate). The extract was filtered and directly injected into a HPLC/time-of-flight mass spectrometry (TOF-MS) system. Quantification was performed using 7H-perfluoroheptanoic acid as internal standard and a calibration standard solution dissolved in sample extract for each matrix type (matrix-matched calibration standard). The method is very time and cost efficient. Except for long-chain compounds and perfluorooctane sulfonamide (which cannot be covered by this method), recoveries were between 60% and 115% and method detection limits were in the range 0.04-1.3 ng/g wet weight. Blank values could be neglected with the exception of perfluorooctane sulfonate (PFOS), perfluorohexanoic acid (PFHxA) and perfluorooctanoic acid (PFOA). One of the major challenges in PFAS analysis is ionisation disturbance by co-eluting matrix in the ion source of the mass spectrometer. Both matrix and analyte specific signal enhancement and suppression was observed and quantified. Repeated extractions (n = 3) gave relative standard deviations (RSD) <35% for all PFAS. Accuracy was examined by comparing the screening method to the generally applied ion pair extraction (IPE) method. PFAS concentration values of a glaucous gull liver sample deviated by less than 30% for the two methods, provided that matrix matched calibration standards were employed in both methods. PMID- 16038212 TI - Fully automated analysis of estrogens in environmental waters by in-tube solid phase microextraction coupled with liquid chromatography-tandem mass spectrometry. AB - A simple, rapid and sensitive method for the determination of five estrogens, estrone, 17beta-estradiol, estriol, ethynyl estradiol, and diethylstilbestrol, was developed using a fully automated method consisting of in-tube solid-phase microextraction (SPME) coupled with liquid chromatography-tandem mass spectrometry (LC/MS/MS). These estrogens were separated within 8 min by HPLC using an XDB-C8 column and 0.01% ammonia/acetonitrile (60/40, v/v) at a flow rate of 0.2 mL/min. Electrospray ionization conditions in the negative ion mode were optimized for MS/MS detection of the estrogens. The optimum in-tube SPME conditions were 20 draw/eject cycles of 40 microL of sample using a Supel-Q PLOT capillary column as an extraction device. The extracted compounds were easily desorbed from the capillary by passage of the mobile phase, and no carryover was observed. Using the in-tube SPME LC/MS/MS method, good linearity of the calibration curve (r > or = 0.9996) was obtained in the concentration range from 10 to 200 pg/mL for all compounds examined. The limits of detection (S/N= 3) of the five estrogens examined ranged from 2.7 to 11.7 pg/mL. The in-tube SPME method showed 34-90-fold higher sensitivity than the direct injection method (5 microL injection). This method was applied successfully to the analysis of environmental water samples without any other pretreatment and interference peaks. Several surface water and wastewater samples were collected from the area around Asahi River, and estriol was detected at 35.7 pg/mL in the effluent of a sewage treatment plant. The recoveries of estrogens spiked into river waters were above 86%, except for estriol, and the relative standard deviations were below 0.9-8.8%. PMID- 16038213 TI - New quantification procedure for the analysis of chlorinated paraffins using electron capture negative ionization mass spectrometry. AB - An improved quantification procedure for the analysis of chlorinated paraffins (CPs) is presented based on electron capture negative ionization mass spectrometry. It compensates differences in response factors between reference CP mixtures and the CP pattern present in environmental samples. The use of a CP standard with a matching degree of chlorination is no longer necessary. It could be shown that the response factors of C10-, C11-, C12- and C13-CP mixtures of both 50 and 60% chlorine content were only slightly influenced by the carbon chain length. A linear correlation (R2 = 0.965) between the total response factor of a CP mixture and its chlorine content was obtained for seven short chain chlorinated paraffin mixtures (SCCP, C10-C13) with different composition and chlorine content (51-69%). Maximum single deviations were <7% for this reference set. It allowed to determine the correct total response factor of the CP composition present in a sample. The deviations were not more than 7-33% for five independent SCCP control samples compared to up to 373% for the conventional procedure. The procedure was tested by quantifying the SCCP and MCCP levels in 10 fish liver samples. The proposed method allowed to compensate the influence of the degree of chlorination of the applied reference standard on the total response factor. PMID- 16038214 TI - Speciation analysis of inorganic arsenic by microchip capillary electrophoresis coupled with hydride generation atomic fluorescence spectrometry. AB - A novel method for speciation analysis of inorganic arsenic was developed by on line hyphenating microchip capillary electrophoresis (chip-CE) with hydride generation atomic fluorescence spectrometry (HG-AFS). Baseline separation of As(III) and As(V) was achieved within 54 s by the chip-CE in a 90 mm long channel at 2500 V using a mixture of 25 mmol l(-1) H3BO3 and 0.4 mmol l(-1) CTAB (pH 8.9) as electrolyte buffer. The precisions (RSD, n=5) ranged from 1.9 to 1.4% for migration time, 2.1 to 2.7% for peak area, and 1.8 to 2.3% for peak height for the two arsenic species at 3.0 mg l(-1) (as As) level. The detection limits (3sigma) for As(III) and As(V) based on peak height measurement were 76 and 112 microg l(-1) (as As), respectively. The recoveries of the spikes (1 mg l(-1) (as As) of As(III) and As(V)) in four locally collected water samples ranged from 93.7 to 106%. PMID- 16038215 TI - Polyacrylamide gel electrophoresis separation and detection of polyamidoamine dendrimers possessing various cores and terminal groups. AB - Detection and separation of polyamidoamine dendrimers possessing various cores and surface groups was studied by polyacrylamide gel electrophoresis. Although many dyes and staining techniques were able to detect dendrimers on polyacrylamide gels, Coomassie Blue was found to be the most sensitive and convenient. Amine and hydroxyl terminated dendrimers were best separated under acidic conditions, while dendrimers with carboxyl surfaces required alkaline buffers. Some dendrimers were very susceptible to diffusion that could occur during their separation, staining or destaining steps. In the absence of an appropriate fixation step, dendrimers could be resolved by using small pore size gels and low voltage or current. Increasing core lengths did not significantly affect migration of a given dendrimer generation but exhibited improved separation and staining characteristics. Polyacrylamide gel electrophoresis was found to be a rapid, inexpensive, and reliable procedure to characterize many different water-soluble dendritic macromolecules. PMID- 16038216 TI - Multi-walled carbon nanotubes as a solid-phase extraction adsorbent for the determination of chlorophenols in environmental water samples. AB - Multi-walled carbon nanotubes (MWCNs) are used as adsorbent for solid-phase extraction (SPE) of several chlorophenols (CPs). CPs were adsorbed on MWCNs cartridge, then desorbed with pH 10.0 methanol, finally determined by HPLC. Under the optimized conditions, detection limits of 0.08-0.8 ng mL(-1) were obtained. The method had been applied to analyze the five CPs in tap water and river water. PMID- 16038217 TI - System constants for the bis(cyanopropylsiloxane)-co-methylsilarylene HP-88 and poly(siloxane) Rtx-440 stationary phases. AB - The solvation parameter model is used to characterize the retention properties of the bis(cyanopropylsiloxane)-co-methylsilarylene, HP-88, and poly(siloxane), Rtx 440, stationary phases over the temperature range 60-140 degrees C. HP-88 is among the most cohesive, dipolar/polarizable and hydrogen-bond basic of stationary phases for open-tubular column gas chromatography. It has no hydrogen bond acidity or capacity for electron lone pair interactions. It exhibits similar selectivity to the poly(cyanopropylsiloxane) stationary phase SP-2340. Rtx-440 is a low-polarity, low-cohesion stationary phase with a moderate capacity for dipolar/polarizable and hydrogen-bond base interactions. It has no hydrogen-bond acidity and possesses weak electron lone pair interactions. It has unique selectivity when compared against a system constants database for 28 common stationary phase compositions. Cluster analysis indicated that the poly(cyanopropylphenyldimethylsiloxane) stationary phase containing 6% cyanopropylphenylsiloxane monomer, DB-1301, the poly(dimethyldiphenylsiloxane) stationary phase containing 20% diphenylsiloxane monomer, Rtx-20, the poly(siloxane) stationary phase of unknown composition, DB-624, and DX-1 [a mixture of poly(dimethylsiloxane) and poly(ethylene glycol) 9:1] are the closest selectivity matches in the database. The selectivity of DB-1301 and Rtx-440 are very similar for solutes with weak hydrogen-bond acidity allowing one stationary phase to be substituted for the other with likely success. For strong hydrogen bond acids, such as phenols, DB-1301 and Rtx-440 exhibit different selectivity. PMID- 16038218 TI - Bleeding stromal tumor in Meckel's diverticulum. PMID- 16038219 TI - Familial tubular breast carcinoma. PMID- 16038220 TI - Hypopituitarism due to hypothalamic B-cell lymphoma. PMID- 16038221 TI - Retroscapular hibernoma. PMID- 16038222 TI - Tracheal nodularity due to tracheobronchopathia osteochondroplastica. PMID- 16038223 TI - Incisional inferior lumbar hernia. PMID- 16038224 TI - Central pontine and thalamic myelinolysis in an alcoholic patient. PMID- 16038225 TI - Trauma-related conductive hearing loss. PMID- 16038226 TI - Lemierre syndrome with jugular thrombophlebitis and cavitary pneumonia. PMID- 16038227 TI - Pulmonary alveolar microlithiasis in two brothers. PMID- 16038228 TI - Cardiac insufficiency due to left arterial benign myxoma. PMID- 16038229 TI - Persistent hypoglossal artery found incidentally. PMID- 16038230 TI - Type I hydatid cyst of the liver: typical MRI features. PMID- 16038231 TI - Ovarian torsion in a 12-year-old girl. PMID- 16038232 TI - Granulocytic sarcoma or chloroma. PMID- 16038233 TI - Paragastric splenosis. PMID- 16038234 TI - Tuberous sclerosis and subependymal giant cell astrocytoma. PMID- 16038235 TI - Cystic tuberculosis of talus. PMID- 16038236 TI - Primary cerebral neuroblastoma. PMID- 16038237 TI - Pulmonary cystic fibrosis with fatty replacement of pancreas. PMID- 16038238 TI - Aggressive lymphoma of the skull in a patient with AIDS. PMID- 16038239 TI - Posttraumatic iliopsoas bursitis. PMID- 16038240 TI - Paget disease of the skull and temporal bone. PMID- 16038241 TI - Sternocostoclavicular hyperostosis in SAPHO-syndrome. PMID- 16038242 TI - Imperforated Cowper's syringocele. PMID- 16038243 TI - Reflections on the ILA African Leprosy Congress. PMID- 16038244 TI - Epidemiological shift in leprosy in a rural district of central India following introduction of multi-drug therapy (April 1986 to March 1992 and April 1992 to March 2002). AB - This study compares the epidemiological pattern of leprosy in pre- (April 1986 to March 1992) and post- (April 1992 to March 2002) multi-drug therapy (MDT) periods by retrospective analysis of 3274 registered leprosy cases in the rural field area of Regional Leprosy Training & Research Institute (RLTRI), situated in Raipur district of Chattisgarh province of Central India. The area has high endemicity for leprosy. In the post-MDT period, prevalence rate (PR) came down to less than 1 in 10, while New Case Detection Rate (NCDR) remained almost static during the two periods. Of the total new registered cases, 30.1% were registered during the pre-MDT period and the remaining 69.9% during the post-MDT period. Comparison of key leprosy variables among new registered cases showed a 2-fold rise in the proportion of MB cases (14.8 versus 27.6%), 3.0% increase in proportion of child cases (15.3 versus 18.6%) and cases with deformity grade II (3.1 versus 5.9%) and 4.0% increase in female proportion (41.4 versus 45.7%) during the post-MDT period. A decline was noted in mean age of registration for both MB (6.4 years) and PB (5.7 years) groups in the post-MDT period. While comparing treatment and outcome related variables, a marked fall of 25.8 months was recorded in treatment duration in the post-MDT period. The defaulter rate came down by 45.0% and relapse rate by more than 12.0% during the same period. The study shows that MDT is effective operationally, but continued ongoing transmission of infection and delayed diagnosis needs corrective action. PMID- 16038245 TI - Curing the stigma of leprosy. AB - The stigma of leprosy is a real phenomenon in many people's lives that affects their physical, psychological, social and economical well-being. There are many causes for this damaging image of leprosy. There is no one easy answer to dispelling this image; it is something that has to be done in partnership with communities and patients. Many papers document the effects of stigma, but few discuss or trial solutions. Education and media campaigns counteract false beliefs about leprosy and raise awareness of new advances in the field. Leprosy care is increasingly provided in an integrated setting showing patients and their communities that leprosy is not a disease apart. Physical and socio-economic rehabilitation is worthwhile in restoring self worth and status in the community and helps patients to find employment. Group counselling can allow those with leprosy to talk about their feelings and experiences to empower one another. Gradually attitudes towards leprosy are changing, but there is still much to be done if the underlying menace of stigma is to be dealt with. We as health professionals must be prepared to make the first move and give that first touch. Certainly more research is needed. In the highly endemic countries the road to elimination may yet be long. Perhaps with effort we will one day be able not only to treat the disease, but also to cure the stigma of leprosy, and make that road an easier one. PMID- 16038247 TI - High prevalence of non-leprotic hypochromic patches among children in a rural area of Mali, West Africa. AB - The identification of one or several hypochromic patches (HP) on the skin is a key stage in the diagnosis of leprosy on dark skin. However, HP are often caused by other disorder than leprosy. A study to determine the prevalence and causes of HP among children was carried out in a rural area of Mali in November 2001. All children under 15 years of age in two villages in an endemic area were screened by two dermatologists. Among the 1729 children seen, HP were identified in 71 patients, with a prevalence of 4.1%. The most common cause of HP was tinea versicolor, which was present in 39.4% of children with HP, followed by pityriasis alba in 31%, naevus achromicus in 24% and vitiligo in 5.6%. No case of leprosy was detected. Our study raises several points with practical consequences for the detection of leprosy cases: the high prevalence of non-leprous HP compared to leprosy, the reliability of the clinical diagnosis of leprosy, and the role of general health care workers in the detection of leprosy cases. Helping those who should be involved in that detection in distinguishing true cases from other hypochromic disorders appears to be a priority. PMID- 16038246 TI - Immunoprophylactic effects of the anti-leprosy Mw vaccine in household contacts of leprosy patients: clinical field trials with a follow up of 8-10 years. AB - We report here a large scale, double blind immunoprophylactic trial of a leprosy vaccine based on Mycobacterium w (Mw) in an endemic area of Kanpur Dehat, Uttar Pradesh, India. A population of 420,823 spread over 272 villages was screened where 1226 multibacillary (MB) and 3757 paucibacillary (PB) cases of leprosy were detected. A total of 29,420 household contacts (HHC) of these patients were screened for evidence of active or inactive leprosy. After exclusion of 1622 contacts for any of the different exclusion criteria, a total of 24,060 HHC could be vaccinated for vaccine or placebo under coding (20,194 administered two doses and 3866 received single dose). The vaccine consisted of 1 x 10(9) heat killed bacilli (Mw) in normal saline for the first dose and half of the first dose, i.e. 5 x 10(8) bacilli for the second dose, given 6 months after the first dose. The placebo consisted of 1/8th dose of the normal dose of tetanous toxoid. Both placebo and vaccine were given under double-blind coding, The contacts were followed up during three surveys at 3, 6 and 9 years after the initial vaccination, for detection of post-vaccination cases (PVCs) and observing any side-effects caused as a result of vaccination. The codes were opened on 24th January 2001, after the analysis of the data following completion of the third and final follow-up survey. When only contacts received the vaccine, Mw vaccine showed a protective efficacy (PE) of 68-6% at the end of first, 59% at the end of the second and 39.3% at the end of the third follow-up survey. When both patients and contacts received the vaccine, the protective efficacy observed was 68%, 60% and 28% at the end of the first, second and third surveys, respectively. When patients, and not the contacts, received the vaccine, a PE of 42.9% in the first, 31% in the second and 3% in the third survey was shown. These results suggest that the vaccination of the contacts is more valuable in achieving the objective of immunoprophylaxis than that of patients, and the vaccine effects are noted maximally in children (as compared to adolescents and adults) who constitute the most responsive group The effect of vaccine is sustained for a period of about 7 8 years, following which there is a need to provide a booster vaccination for the sustained protection. PMID- 16038248 TI - Report on the first meeting of the IDEAL (Initiative for Diagnostic and Epidemiological Assays for Leprosy) consortium held at Armauer Hansen Research Institute, ALERT, Addis Ababa, Ethiopia on 24-27 October 2004. PMID- 16038249 TI - How can adherence with multi-drug therapy in leprosy be improved? PMID- 16038250 TI - Leprosy and HIV co-infection in five patients. AB - In a retrospective study, five patients are reported who suffered from a Mycobacterium leprae/HIV co-infection and were treated for their HIV infection with HAART. In four patients, this revealed their leprosy and induced a type I leprosy reaction. Two patients who were lepromin negative at diagnosis were retested after about 1 year of anti-retroviral treatment, and found to be positive. PMID- 16038251 TI - Aplastic anaemia associated with multidrug therapy (dapsone, rifampicin and clofazimine) in a patient with lepromatous leprosy. PMID- 16038252 TI - Lichen scrofulosorum in a patient with lepromatous leprosy after BCG immunotherapy. AB - Lichen scrofulosorum is a rare form of tuberculid seen in children and young adults. The cutaneous lesions are typically symptomless papular eruptions, associated with a strong Mantoux reaction, tuberculosis of lymph nodes and/or other organs or rarely following BCG vaccination. We describe an unusual case of occurrence of lichen scrofulosorum following BCG immunotherapy in a patient with lepromatous leprosy. PMID- 16038253 TI - Leprosy acquired by inoculation from a knee injury. AB - This case study reports on the development of clinical leprosy in a young Caucasian female from a non-endemic country who contracted the disease while living in a leprosy endemic country. In the presentation and discussion, some relevant factors will be reviewed and discussed that may play a role in the transmission, susceptibility and clinical development of the disease. PMID- 16038254 TI - Independent evaluation of GAEL. PMID- 16038255 TI - Bigger may not always be better: giant borderline tuberculoid leprosy lesion. PMID- 16038256 TI - [Disease gene identification: opportunities and challenges]. AB - The recent achievements of the Human Genome Project make it increasingly feasible to determine the genetic basis of human diseases, especially complex traits. Genomics will provide powerful means to discover hereditary elements that interact with environmental factors leading to diseases. However, the expected transformation toward genomics-based medicine will occur over decades, which requires the joint efforts of many scientists and physicians. Such transformation provides both opportunities and challenges to everyone involved in this field. PMID- 16038257 TI - [GNU Pattern: open source pattern hunter for biological sequences based on SPLASH algorithm]. AB - OBJECTIVE: To construct a high performance open source software engine based on IBM SPLASH algorithm for later research on pattern discovery. METHODS: Gpat, which is based on SPLASH algorithm, was developed by using open source software. RESULTS: GNU Pattern (Gpat) software was developped, which efficiently implemented the core part of SPLASH algorithm. Full source code of Gpat was also available for other researchers to modify the program under the GNU license. CONCLUSION: Gpat is a successful implementation of SPLASH algorithm and can be used as a basic framework for later research on pattern recognition in biological sequences. PMID- 16038258 TI - [Altered expression of the HSD17B4 gene in esophageal squamous cell carcinoma and loss of heterozygosity analysis]. AB - OBJECTIVE: To investigate the alteration of the gene HSD17B4 in esophageal squamous cell carcinoma and its potential significance. METHODS: The mRNA expression and loss of heterozygosity (LOH) of HSD17B4 in 40 primary esophageal tumors were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and microsatellite analysis with the intragenic marker D5S1384 of the gene. RESULTS: The frequencies of allelic loss of D5S1384 and the rate of down regulation of gene HSD17B4 were 46.2% and 62.5%, respectively. CONCLUSION: HSD17B4 may be a candidate tumor suppressor gene associated with esophageal squamous cell carcinoma. PMID- 16038259 TI - [Screening susceptibility genes of type 2 diabetes in Chinese population by single nucleotide polymorphism analysis]. AB - OBJECTIVE: To search for the susceptibility variant (s) of type 2 diabetes in the susceptible regions on chr.1 (1p36.23-36.33, 1q24.3-25.1, and 1q42.12-42.13) by genotyping SNP markers in case-control DNA samples and identifying the haplotype associated with type 2 diabetes. METHODS: Totally 124 SNPs in 33 candidate genes in the mapped regions were chosen from public SNP data or identified by sequencing the samples that were used to search for SNP locus. Sequencing method was used to genotype the loci for 236 sporadic type 2 diabetes patients and 152 normal subjects in Northern Han Chinese population. The haplotypes with significant difference were further analyzed. RESULTS: Of 124 SNPs successfully typed, 4 SNPs that showed association with diabetes status were found: rs203849 (P=0.005, OR=1.60) and rs203826 (P=0.016, OR=1.60) located in sAC gene, rs7535528 (P=0.028, OR=1.45) located in PANK4, rs884363 (P=0.043, OR=1.37) located in CASP9 gene. In addition, the frequencies of two combination types from these 4 SNP genotypes were significantly different between case and control groups (P < 0.001). Furthermore, four haplotypes associated with diabetes were found in haplotype analysis of sAC gene. CONCLUSION: sAC, PANK4, and CA SP9 may be associated with type 2 diabetes in Han population in north China, and it seems that the synergetic effect of these genes is responsible for the development of type 2 diabetes. PMID- 16038260 TI - [Nuclear localization region in soluble adenylyl cyclase]. AB - OBJECTIVE: To locate the region responsible for nuclear localization of protein sAC. METHODS: The eukaryotic expression vector of vairous sAC deletion mutants were transfected into Hela cells. The localization of each mutant was observed using confocal microscope. RESULTS: For some mutants, the localization of sAC changed. Deletion of some region made it unable to locate in the nuclear. CONCLUSION: It is possible to figure out that the nucleotide region (739-1038 and 1045-1261) take charge of nuclear localization of sAC. PMID- 16038261 TI - [NQ01 gene polymorphism C609T associated with an increased risk for cognitive dysfunction and sporadic Alzheimer's disease in Chinese]. AB - OBJECTIVE: To investigate the association between the C609T polymorphism of NADP (H): quinoneoxidoreductase 1 (NQ01) gene and decreased cognitive function and sporadic Alzheimer's disease (AD) in a community cohort. METHODS: Polymerase chain reaction (PCR), denaturing high performance liquid chromatography (DHPLC) and sequencing were used to determine the genotype of NQ01 in 110 subjects without cognitive dysfunction, 21 with cognitive dysfunction, and 65 AD patients from a community cohort. RESULTS: Significantly different distributions of C/T and T/T genotypes were found between MMSE normal and abnormal subjects (OR=2.8, 95%CI 0.96-8.18, P=0.024), and between AD patients and healthy controls (OR=3.27, 95% CI 1.54-6.94, P=0.001), respectively. The frequencies of T allele of NQ01 C609T were significantly higher in MMSE abnormal subjects and AD patients (P=0.034 and 0.005) as compared to normal controls. CONCLUSION: The C609T polymorphism of NQ01 gene may be a genetic risk factor for cognitive dysfunction and sporadic AD in Chinese population. PMID- 16038262 TI - [Relationship between congenital long QT syndrome and Brugada syndrome gene mutation]. AB - OBJECTIVE: To investigate the molecular pathology in families with long QT syndrome (LQTS) including Jervell-Longe-Nielsen syndrome (JLNS) and Romano-ward syndrome (RWS) and Brugada syndrome (BS) in Chinese population. METHODS: Polymerase chain reaction and DNA sequencing were used to screen for KCNQ1, KCNH2, KCNE1, and SCN5A mutation. RESULTS: We identified a novel mutation N1774S in the SCN5A gene of the BS family, a novel mutation G314S in a RWS family which had also been found in Europe, North America, and Japan, and a single nucleotide polymorphisms (SNPs) G643S in the KCNQ1 of the JLNS family. In this JLNS family, another heterozygous novel mutation in exon 2a was found in KCNQ1 of the patients. CONCLUSION: New mutations were found in our experiment, which expand the spectrum of KCNQ1 and SCN5A mutations that cause LQTS and BS. PMID- 16038263 TI - [Recombinant human pigment epithelium-derived factor inhibits the proliferation of the endothelial cells from blood vessels]. AB - OBJECTIVE: To express and purify the recombinant human pigment epithelium-derived factor (PEDF) which inhibits the proliferation of the endothelium cells from blood vessel in E.coli. METHODS: PEDF gene was inserted into the prokaryotic expression vector pGEX-4T-2. The recombinant protein PEDF was expressed in E.coli BL-21, and purified by the GST Sepharose 4B affinity column. The recombinant human PEDF protein was identified by Western blot and mass spectrum. The biological activity of the recombinant human PEDF protein was measured by using MTT. RESULTS: The 46 kDa recombinant human PEDF protein was obtained. It significantly inhibited the proliferation of the human umbilical vein cell line HUVEC. CONCLUSION: The recombinant human PEDF with anti-angiogenesis activity protein may be successfully purify through prokaryotic expression. PMID- 16038264 TI - [Inhibition of multidrug resistance related P-gp expression in human neuroblastoma by antisense peptide nucleic acid]. AB - OBJECTIVE: To investigate the efficiency of a peptide nucleic acid (PNA) delivery system by using liposome via PNA-DNA hybrids and to test the inhibitive action of antisense PNA on expression of multidrug resistance (MDR) related P-glycoprotein (P-gp) in human neuroblastoma cell line SK-N-SH. METHODS: Two antisense PNAs were designed targeting at MDR-1 mRNA and then combined with partially complement DNAs respectively. The hybrids were delivered into cells using cationic liposome. The transfection efficiency, expression of P-gp and MDR-1 mRNA, intracellular adarimycin (ADM) were measured by flow cytometry, reverse transcription polymerase chain reaction (RT-PCR), and high performance liquid chromatography (HPLC). RESULTS: Transfection of PNA increased the cell average fluorescence intensity significantly and the extent of increase was dependent on the concentration of PNA. After being transfected by both PNAs, P-gp expression of SK N-SH cells decreased significantly and the intracellular ADM level was increased by about 3 times. The level of MDR-1 mRNA expression slightly decreased after transfection, but no statistical significance was observed. CONCLUSIONS: PNA can be delivered into tumor cells in form of PNA-DNA hybrids by cationic liposome. Properly designed antisense PNA can inhibit MDR related P-gp expression of SK-N SH cells efficiently and specifically. PMID- 16038265 TI - [Differential gene expression in nasopharyngeal carcinoma cell with reduced and normal expression of 6A8 alpha-mannosidase]. AB - OBJECTIVE: To detect the differential display of mRNA expression between human nasopharyngeal carcinoma cell CNE-2L2 with reduced malignancy caused by transduction of a DNA antisense to 6A8 alpha-mannosidase cDNA (AS cell) and the wild type cell (W cell). METHODS: Differential display of mRNA expression was analyzed using DNA microarray analysis. The datasets were confirmed by Northern blotting and RT-PCR. RESULTS: Out of the 1069 genes analyzed, 34 genes were up regulated in AS cells relative to W cells. Conversely, 42 genes were down regulated. The genes, up-regulation of which might have suppressive effect on tumor malignant behaviors, were P130 mRNA for 130K protein, TGF-betaIIR alpha, GABBR1, TGFBR1, TNFAIP1, STANIN, E-CADHERIN, CTNNA1 and 2, RFX2, TMPO, etc. The genes, down-regulation of which might have suppressive effect on tumor malignant behaviors, were CD44, NDRG1, TGFB1, RPS5, LEGUMAIIN, CBS, CD59, SNRPA1, etc. The microarray datasets were confirmed by Northern blot and RT-PCR analysis. CONCLUSIONS: In comparison to the W cell, AS cell has up-regulation of 34 genes and down-regulation of 42 genes. Changes of the gene expression may play a role in the malignancy reduction of AS cell. PMID- 16038266 TI - [Effect of calmodulin antagonist EBB on invasion of human fibrosarcoma cell HT1080]. AB - OBJECTIVE: To investigate the potential effect of EBB, a calmodulin antagonist, on invasion of human fibrosarcoma cells HT1080. METHODS: The antitumor effect of EBB was assessed by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Activities of matrix metalloproteinase (MMP)-2 and MMP-9 were measured by Zymogrophy analysis. The mRNA levels, of MMP-2, MMP-9, and tissue inhibitor of metalloproteinases (TIMP)-1 were evaluated by reverse transcriptionpolymerase chain reaction (RT-PCR). Transwell chamber assay was applied to measure the effect of EBB on the invasion of HT1080 cells. RESULTS: Calmodulin antagonist EBB inhibited the proliferation of HT1080 cells with an IC50 of (8.2 +/- 1.2) microg/ml. EBB down-regulated the activities of MMP-2 and MMP-9, and down-regulated the mRNA levels of MMP-2 and MMP-9, while up-regulated the mRNA levels of TIMP-1. The invasive ability of HT1080 cells was decreased to (31.13 +/- 2.265)%, (59.91 +/- 2.566)%, and (71.58 +/- 0.5960)% after exposure of the cells with 2, 5, and 10 microg/ml EBB, respectively. CONCLUSION: Treatment with calmodulin antagonist EBB is effective in suppressing tumor invasion. The possible mechanism is the down-regulation of MMPs. PMID- 16038267 TI - [Ex vivo expansion of Valpha24 natural killer T cells with alpha galactosylceramide]. AB - OBJECTIVE: To evaluate the method for expanding Valpha24 natural killer T (NKT) cells with alpha-galactosylceramide (alpha-GalCer) ex vivo. METHODS: Mononuclear cells (MNCs) isolated from adult peripheral blood or umbilical cord blood (UCB) were divided into three groups. In Group A1 (n = 5), CD34+ progenitorderived dendritic cells were differentiated in a cytokine-supplemented culture system from cord blood and acted as antigen presenting cells (APC) to induce the expansion of cord blood Valpha24 NKT cells in presence of alpha-GalCer; in Group A2 (n = 5), adult peripheral monocyte-derived dendritic cells (Mo-DC) were used as APC to induce the expansion of adult peripheral NKT cells in presence of alpha GalCer; whereas in Group B (n = 16), alpha-GalCer was added into adult peripheral MNCs culture system without additional DCs. Cytokine-produce were measured by ELISA, and NKT cells' proliferation ability, cytotoxicity, and suppressive effect on mixed lymphocyte reaction (MLR) were examined by MTT assays. RESULTS: Valpha24 NKT cells in Group A1, A2, and B were expanded up to 128 (95-207), 250.5 (179.6 790.6), and 326 (101-2 136) -fold by day 12, respectively. Adult NKT cells expanded in Group B were markedly better than those in Group A1 (P = 0.038). When stimulating by PMA, the NKT cells had a 3-day stimulate index of 1.80 +/- 0.41; and the secretion ratio of IL-4 to IFN-gamma of UCB or adult peripheral blood NKT cells were 0.30 +/- 0.13 and 0.28 +/- 0.18; and the ex vivo antitumor effect of expanded NKT cells were found in cell line HL60, KG1a, and Raji except for K562; and the suppressive effect of expanded NKT cells or the culture supernatant on MLR were confirmed. CONCLUSIONS: Alpha-GalCer can facilitate the rapid shorttime expansion of Valpha24 NKT cells in presence of IL-2 and IL-15. These expanded NKT cells, kill tumor cell lines, and inhibit can massively excret IL-4 and IFN-gamma allogeneic T-cell response. PMID- 16038268 TI - [Relationship between the increase of hepatic D-bifunctional protein activity and bile acid biosynthesis in rats]. AB - OBJECTIVE: To determine the physiological role of D-bifunctional protein (DBP) in bile acid biosynthesis through investigating the effect of increasing activity of DBP on bile acid biosynthesis. METHODS: Twenty male Wistar rats were divided into two groups: diethylhexyl phthalate (DEHP) group (n = 10) and control group (n = 10). Serum triglyceride, total cholesterol, hepatic DBP activity, and fecal bile acids were assayed. The mRNA levels of hepatic peroxisome proliferator-activated receptor alpha (PPARalpha), DBP, and cholesterol 7alpha-hydroxylase (CYP7A1) were detected by RT-PCR. RESULTS: Compared with control group, serum triglyceride level was decreased significantly and PPARalphamRNA level was increased significantly in DEHP group (P < 0.01). Together with a sharp induction of DBP mRNA expression and DBP activity in DEHP group (P < 0.01), the levels of CYP7A1 mRNA and fecal bile acids were significantly increased by 1.9 times and 1.6 times respectively compared to control group (P < 0.01). There was a significantly positive correlation between DBP mRNA level or DBP activity and CYP7A1 mRNA level (r = 0.89, P < 0.01; r = 0.95, P < 0.01). CONCLUSION: The up-regulation of DBP mRNA and activity in liver can result in the increase in CYP7A1 mRNA expression and bile acid biosynthesis, suggesting that DBP may be involved in bile acid biosynthesis together with CYP7A1. PMID- 16038269 TI - [Altered expression of vascular endothelial growth factor and its receptors in transdifferentiated human proximal tubular epithelial cells induced by transforming growth factor beta1]. AB - OBJECTIVE: To examine the expression of vascular endothelial growth factor (VEGF) and its receptors (VEGFR1, VEGFR2) in transdifferentiated human proximal tubular epithelial (HK-2) cell induced by transforming growth factor beta1 (TGFbeta1). METHODS: The transdifferentiation of HK-2 cells was detected by evaluation of expression of alpha-SMA by cytoimmunochemistry and RT-PCR. The VEGF mRNA was evaluated with RT-PCR. The secreted VEGF in the culture media was measured with ELISA. The cellular VEGF, VEGFR1, and VEGFR2 were measured with Western blot. RESULTS: The immunostain of alpha-SMA were positive in HK-2 cell induced by TGFbeta1 at the concentration of 5 and 8 ng/ml for 72 h. The expression of alpha SMA mRNA was induced by TGFbeta1 in concentration- and time-dependent manners. The expressions of mRNA and protein of VEGF were upregulated by TGFbeta1 at the concentration of 0.1 and 1 ng/ml for 72 h and at the concentration of 8 ng/ml for 12 h and 24 h when compared with the control. But expressions of mRNA and protein of VEGF were downregulated by TGFbeta1 at the concentration of 3, 5, and 8 ng/ml for 72 h and at the concentration of 8 ng/ml for 36, 48, and 72 h, respectively. Meanwhile, Protein levels of VEGFR1 and VEGFR2 were upregulated by TGFbeta1 in concentration- and time- dependent manners. CONCLUSIONS: Increased expression of VEGFR1 and VEGFR2 and two-phase change in VEGF expression occurred in the process of tubular epithelial transdifferentiation induced by TGFbeta1. Reduced expression of VEGF may contribute to tubular epithelial transdifferentiation in a vicious circle. PMID- 16038270 TI - [Effect of the positive sera of autoantibodies against the human beta1 adrenoceptor from patients with congestive heart failure on activity of L-type Ca2+ channel in guinea pig cardiac myocytes]. AB - OBJECTIVE: To investigate the effect of the positive sera of autoantibodies against the human beta1-adrenoceptor from patients with congestive heart failure on activity of L-Type Ca2+ channel in guinea pig cardiac myocytes. METHOD: Using whole cell patch-clamp technique, we quantitatively researched the ionic intensity and density of L-type Ca2+ channel (ICa-L). RESULTS: The beta adrenocepter agonist isoprenaline increased the ICa-L peak intensity and density from (997.09 +/- 227.5) pA and (8.20 +/- 0.86) pA/pF to (2241.01 +/- 348.5) pA and (18.98 +/- 1.18) pA/pF, respectively (P < 0.01). The positive sera of autoantibodies against the beta1-adrenoceptor could also increase ICa-L peak intensity and density from (963.57 +/- 207.56) pA and (8.14 +/- 0.72) pA/pF to (1382.41 +/- 241.36) pA and (11.70 +/- 1.03) pA/pF (P < 0.01). Esmolol, a beta1 adrenoceptor antagonist blocked these effects of isoprenaline and autoantibodies. CONCLUSIONS: Human cardiac positive sera of autoantibodies against the beta1 adrenoceptor has an isoproterenol-like effect on cardiac myocytes receptor. It may participate in the pathophysiologic process of cardiac myocytes. PMID- 16038271 TI - [Clinical application of portable spirometry in asthma]. AB - OBJECTIVE: To investigate the clinical applications of portable spirometry in asthma. METHODS: Twenty patients with asthma were recruited from Peking Union Medical College Hospital. Flow-volume loop, simultaneous asthma symptoms, and mood were monitored three times a day for consecutive 14 days. RESULTS: In patients with a normal daytime spirometry, marked decline of forced expiratory volume in one second (FEV1) and peak expiratory flow (PEF) were observed at night and/or in the early morning. A within subject correlation analysis between FEV1, PEF, and asthma symptoms showed that the correlation between symptoms and airway obstruction was found only in seven out of twenty patients (35%). Four patients (20%) reported many symptoms with nearly normal portable spirometry. Accordingly, their symptoms were not correlated with FEV1 and PEF. This group of patients was defined as over-perceivers. On the contrary, another two patients (10%) did not report any symptoms while obvious airways obstruction was recorded by a portable spirometry. These patients were defined as under-perceivers. CONCLUSIONS: Dynamic monitoring of flow-volume loop with a portable spirometry is more accurate than routine lung function test in assessment of asthma severity. In addition, combined with simultaneous monitoring of symptoms, it would be of particularly helpful in identifying two specific types of asthma patients, e.g. over perceivers and under-perceivers. PMID- 16038272 TI - [Expression of plasminogen activator inhibitor-1 and tissue plasminogen activator in the kidney of KKAy mice with type 2 diabetes]. AB - OBJECTIVE: To study the role of plasminogen activator inhibitor-1 (PAI-1) and tissue plasminogen activator (tPA) in the accumulation of extracellular matrix (ECM) in the kidney of KKAy mice with type 2 diabetes. METHODS: KKAy mice, a type 2 diabetic animal model, and C57BL-J mice were sacrificed at 16, 20, and 24 weeks of age, respectively. The local expression of renal laminin was analyzed with immunohistochemistry. Chromogenic substance was used to show the activity of PAI 1. The mRNA expression of tPA was determined by RT-PCR. The mRNA expression of PAI-1 was measured by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Laminnin expression was significantly increased in all age groups of KKAy mice. The tPA mRNA was significantly lower than that in C57BL mice, especially at the age of 16w (only 47%). Otherwise the PAI-1 mRNA expression was remarkably up-regulated than that in C57BL mice. CONCLUSION: In type 2 diabetes KKAy mice, the accumulation of ECM may be associated with the abnormal expression of PAI-1/tPA mRNA. PMID- 16038273 TI - [Meta-analysis on the relationship of chlamydia pneumonia infection and coronary heart disease]. AB - OBJECTIVE: To determine the relationship of chlamydia pneumonia (CP) infection and coronary heart disease (CHD). METHODS: Studies published before 2004 that had reported on correlations between CHD and CP were collected. Studies were sorted into four sorts according to different testing methods. We determined to use either fixed effect model or random effect model according to the result of heterogeneity test. Meta-analysis was done by RevMan 4.2. RESULTS: Totally 29 studies were included into the review. The odd ratio (OR) to review all literature enzyme-linked immunosorbent assay (ELISA), dot-immunogdd filtration assay (DIGFA), and micro-immunofluorescence (MIF) was 4.00 (3.07, 5.22), 2.86 (1.89, 4.32), 14.89 (1.46, 151.57), and 4.46 (3.15, 6.32). CONCLUSION: CP infection is associated with CHD, CP infection may be one of the risk factors of CHD. PMID- 16038274 TI - [Mechanism of alopecia in patients with paraneoplastic pemphigus]. AB - OBJECTIVE: To investigate the relationship between the levels of antidesmoglein (DSG) 1, 3 antibodies in the sera of patients with paraneoplastic pemphigus (PNP) and alopecia. METHODS: Sera from PNP patients, bullous pemphigoid patients, and normal healthy subjects were collected and 2 tissue samples from 2 healthy scalps were resected. Anti-DSG 1, 3 antibodies in the sera of PNP patients were detected by enzyme-linked immunosorbent assay (ELISA). Indirect immunofluorescent assay was used to detect whether the antibodies in the sera of PNP patients binds with the follicular epithelium of normal healthy scalp. RESULTS: Anti-DSG3 autoantibody was strongly positive and anti-DSG1 weakly positive in one patient, while both two antibodies were negative in the other patient. Their sera could bind to keratinocytes and follicular epithelium in human scalp. Immunofluorescent signals were found on the intercellular epidermal cell surface and outer root sheath of the follicular epithelium. However, the immunofluorescent signals in the section incubating with serum of bullous pemphigoid were only found on basal membrane zone. No signals were found in the section incubating with normal healthy serum. CONCLUSION: Alopecia in PNP patients are correlated with the anti DSG3. PMID- 16038275 TI - [Orthognathic surgery and distraction osteogenesis for treatment of obstructive sleep apnea hypopnea syndrome]. AB - OBJECTIVE: To discuss the indication and protocol of surgical therapy when treating mandibular micrognathism accompanying obstructive sleep apnea-hypopnea syndrome (OSAHS) using distraction osteogenesis and orthognathic surgery. METHODS: A total of 17 patients with mandibular micrognathism accompanying moderate to severe OSAHS, aged 11 to 59 years, 15 men and 2 women, were reviewed. Four of them were treated with orthognathic surgery, 5 of them were treated with distraction osteogenesis, and the other 8 patients were treated with an integrated procedure combining distraction osteogenesis with orthognathic surgery. Cephalometric analysis and polysomnography studies were obtained pre- and postoperatively. RESULTS: SNB angle changed from 64.6 degrees to 71.9 degrees, post airway space (PAS) from 5.4mm to 13.2 mm, apnea and hypopnea index (AHI) from 58.4 to 7.6, lowest saturation of oxygen (LSAT) from 66% to 87%. All the differences showed statistical significance (P < 0.001). The average mental horizontal advancement (MHA) was 14.3 mm and its correlation coefficients with deltaSNB, deltaPAS, deltaAHI, and deltaLSAT were 0.36, 0.62, 0.34, and -0.14, respectively. CONCLUSIONS: Both distraction osteogenesis and orthognathic surgery can be effectively used to treat patients with mandibular micrognathism accompanying OSAHS with slightly different indications. A combination of these two operations may be preferred. PMID- 16038276 TI - [Validity and reliability of the Demirjian distinguish software on estimating dental age]. AB - OBJECTIVE: To study the validity and reliability of the Demirjian distinguish software on dental age estimation. METHOD: The dental age of 60 children were estimated with both the software three times and manual measures. RESULTS: The correlation coefficients of dental age estimated by the software and by manual measure was 0.974 (female) (P > 0.1) and 0.970 (male) (P > 0.05); the coefficients of interclass correlation of each dental age estimated by the software was 0.977 (female) (P > 0.1) and 0.977 (male) (P > 0.05). CONCLUSION: Demirjian distinguish software has high validity and reliability in estimating dental age. PMID- 16038277 TI - [Surgical treatment of aortic regurgitation caused by Behcet's disease]. AB - OBJECTIVE: To summarize operational effect with surgical treatment of aortic regurgitation caused by Behcet's disease and discuss relevant surgical techniques for treatment of these conditions. METHODS: Eight patients with aortic regurgitation secondary to Behcet's disease and received surgery between April 1997 and August 2003 were retrospetively analyzed. Among them, two patients had their aortic valves replaced in other hospital before admitted to our hospital where one undertook aortic valve replacement (AVR), and the other undertook aortic root replacement (ARR). In six patients who were initially treated in our hospital, the surgical procedures for aortic regurgitation included AVR in three patients and ARR operation in other three patients in whom Bentall-type operation was conducted in two patients and Cabrol-type operation in one. RESULTS: One patient died during hospital stay. The follow-up periods ranged from 3 months to 36 months. In five patients with prosthetic valve detachment or suture detachment, redo homograft replacement was required in one patient and redo AVR in 3, one patient had redo AVR twice, and the remaining one patient had no surgery at present. Three patients primarily operated by ARR operation have no complications. CONCLUSIONS: The rate of prosthetic valve detachment is high in patients with Behcet's disease. ARR should be a first-line therapy for operation promised these patients. PMID- 16038278 TI - [Initial radiographic manifestations in patients with SARS]. AB - OBJECTIVE: To observe initial radiographic manifestations and evaluate its value for the diagnosis and differential diagnosis in patients with severe acute respiratory syndrome (SARS). METHODS: Initial chest radiographs of 79 patients, 15 to 69 years old (mean age 34.1 years, 40 men and 39 women), were retrospectively reviewed in the study. For all patients, average temperature was 38.8 degrees C and mean time from onset of the fever to the time of initial radiographs taken was 5.8 days. Location, abnormal distribution, appearance of lung abnormalities, effusion of pericadiaum and pleura, and lymphadenopathy were observed. Abnormal distribution and associated findings were compared. Differences of mean age, temperature, and the time were statistically analyzed between patients with and without abnormal findings. RESULTS: Initial chest radiographs were normal in 36 of 79 patients (45.5%). Abnormal radiographs in 43 patients (81.3%) showed exudative changes. No significant difference of mean age and day (form onset of the fever to the time of initial radiographs taken) was found between the patients with and without abnormal initial chest radiographs (P > 0.05). No lymphadenopathy, effusion of pericadiaum and pleura were found on the initial chest radiographs. CONCLUSIONS: The main abnormal radiographic appearance of lung in patients with SARS is exudative changes. SARS patients with normal initial chest radiographs are not uncommon and abnormal radiographic findings may appear to be late after onset of fever. Therefore, more attention should be paid in the clinical diagnosis of SARS. PMID- 16038279 TI - [Treatment of ureteral calculi with ballistic lithoclast]. AB - OBJECTIVE: To evaluate the availability of ballistic lithoclast in treatment of ureteral calculi. METHODS: Totally 625 patients with ureteral calculi were treated with ballistic lithoclast by rigid ureteroscopy. RESULTS: Totally 610 patients were treated successfully by ballistic lithoclast with a cure rate of 97.6% (90.3% in upper ureter, 96.8% in middle ureter, 100% in lower ureter). The average time of lithoclast was (5.0 +/- 2.8) minutes and the stone-free rate was 99.2%. CONCLUSION: Ballistic lithoclast combined with rigid ureteroscopy is safe and effective in treating ureteral calculi. PMID- 16038280 TI - [Modified dacryocystorhinostomy]. AB - OBJECTIVE: To introduce a novelty-modified dacryocystorhinostomy. METHODS: With the innovated scalpels, 11 patients (13 eyes) suffering from chronic dacryocystitis were performed with the newly modified surgical procedures derived from traditional dacryocystorhinostomy. The efficacy, efficiency, and complications were observed. RESULTS: Thirteen operations for 11 patients were performed successfully and efficiently. The wound was during operation neat and with a slight hemorrhage which could be stanched easily. No serious complications were observed intra-operatively. The average time of follow-up was 7.8 months (6 12 months). All the dacryocysto-rhinal canals remained unblocked. CONCLUSIONS: The modified dacryocystorhinostomy with the help of innovated mucous scalpel has high success rate, high efficiency and low complications. It deserves wide application as the first choice for chronic dacryocystitis. PMID- 16038281 TI - [Expression of galectin-3 in invasive prolactinomas]. AB - OBJECTIVE: To investigate the expression of galectin-3 (Gal-3) in prolactinomas. METHODS: Expressions of Gal-3 were evaluated by immunohistochemistry using polyclonal antibody in 16 invasive prolactinomas and 16 prolactinomas. RESULTS: Gal-3 was expressed both in invasive prolactinomas and noninvasive prolactinomas while significantly higher expression seen in the invasive prolactinomas (P < 0.05). CONCLUSION: Gal-3 expression may be used as a useful indicator to determine the invasiveness and prognosis of prolactinomas. PMID- 16038282 TI - [Research development of Mendelian inherited diabetes]. AB - Diabetes mellitus is a chronic syndrome of abnormal metabolism, determined by interaction of multifactorial genetic and environmental factors. Some specific types of diabetes, such as MODY, Leprechaunism, lipoatrophic diabetes, and Rabson Mendenhall syndrome, are monogenic forms of diabetes and are inherited as a Mendelian pattern. The article reviews the research development of these Mendelian inherited diabetes will be reviewed. PMID- 16038283 TI - [Advances in the studies on the molecular and genetic aspects of epilepsy]. AB - Epilepsy is one of the most common and debilitating neurological diseases that affects more than 40 million people worldwide. Genetic factors contribute to the pathogenesis of epilepsy. Molecular genetic studies have identified 15 disease causing genes for epilepsy. The majority of the genes encode ion channels, including voltage-gated potassium channels KCNQ2 and KCNQ3, sodium channels SCN1A, SCN2A, and SCN1B, chloride channels CLCN2, and ligand-gated ion channels CHRNA4, CHRNB2, GABRG2, and GABRA1. Interestingly, non-ion channel genes have also been identified as epilepsy genes, and these genes include G-protein-coupled receptor MASS1/VLGR1, GM3 synthase, and proteins with unknown functions such as LGI1, NHLRC1, and EFHC1. These studies make genetic testing possible in some patients, and further characterization of the identified epilepsy genes may lead to the development of new drugs and new treatments for patients with epilepsy. PMID- 16038284 TI - [Genes and their functional mechanisms in the pathogenesis of muscular dystrophy]. AB - Muscular dystrophy (MD), a group of inherited disorders characterized by progressive skeletal muscle wasting and weakness, can be classified into several groups according to Mendelian inheritance patterns and clinical features. Many genes related to MD have been identified and cloned by genetic linkage analysis and positional cloning strategy. Our understanding of the molecular mechanisms giving rise to muscular dystrophy have made a progress by the functional analysis of proteins encoded by candidate genes for MD. This article reviews genes and their functional mechanisms in the pathogenesis of muscular dystrophy. PMID- 16038285 TI - [Advances in molecular genetics of schizophrenia]. AB - Schizophrenia (MIM 181500) is a complex disorder affecting approximately 1% of the population worldwide. Epidemiologic evidences, together with recent linkage and association studies, have clearly demonstrated the high heritability of schizophrenia (up to 80%). Uncovering the genetic mechanism of schizophrenia has became one of the greatest challenges for both psychiatry and genetics. In recent years, remarkable advances in the genetics of this disorder has been achieved with the rapid growth of human genome information and experiment technologies. Several candidate genes within some of the best-supported linkage regions have been reported and, more importantly, replicated. Moreover, these genes present a significant connection in the signaling pathways implicated in the development of schizophrenia, especially NMDA receptor-mediated glutamate transmission. In this review, we summarize the recent advances in the genetics of schizophrenia, focusing particularly on linkage disequilibrium analysis and the latest understanding of the neurobiology of the disorder. PMID- 16038286 TI - [Research status of genodermatoses in China]. AB - Among over 300 genodernatoses, causative genes have been identified in 170 monogenetic diseases, while gene mapping has been performed in over 100 monogenetic and polygenctic skin diseases. Researches in genodermatoses has rapidly advanced in China in recent ten years. The causative genes involved in multiple familial trichoepithelioma and primary erythermalgia have been found. Two independent genome-wide scans with DNA markers have been performed to detect genetic linkage related to psoriasis and vitiligo. In this review article, we summarize these most recent findings. PMID- 16038287 TI - Csaba Horvath and preparative liquid chromatography. AB - Few chromatographers have been interested in furthering preparative liquid chromatography. The pioneers, Tswett, Kuhn and Lederer, A.J.P. Martin, Tiselius, isolated fractions but as an intermediate step in the analysis of their samples. The progress in electronics and sensors, and in their miniaturization has lead to the paradoxical situation that the analysts never see the transient pure fractions that their detector quantitates. Yet, over the last 25 years, preparative liquid chromatography has become an important industrial process for the separation, the extraction, and/or the purification of many pharmaceuticals or pharmaceutical intermediates, including pure enantiomers, purified peptides and proteins, compounds that are manufactured at the relatively large industrial scale of a few kilograms to several hundred tons per year. This development that has strongly affected the modem pharmaceutical industry is mainly due to the pioneering work of Csaba Horvath. His work in preparative HPLC was critical at both the practical and the theoretical levels. He was the first scientist in modem times to pay serious attention to the relationships between the curvature of the equilibrium isotherms, the competitive nature of nonlinear isotherms, and the chromatographic band profiles of complex mixtures. The thermodynamics of multi-component phase equilibria and mass transfer kinetics in chromatography attracted his interest and were the focus of ground-breaking contributions. He investigated displacement chromatography, an old method invented by Tiselius that Csaba was first to implement in HPLC. This choice was explained by the essential characteristic of displacement chromatography, in that it delivers fractions that can be far more concentrated than the feed. Remarkably, once the basics of nonlinear chromatography had been mastered in his group, most of the applications that were studied by his coworkers dealt with peptides of various sizes and with proteins. Thus, all the applications of preparative HPLC in the biotechnologies derive directly from Csaba's work. Although displacement did not pan out as a general method, the reasons are related more to practical constraints of the production of pharmaceuticals and to the long period of cheap energy that might be ending now. This report reviews Csaba's work in nonlinear chromatography. PMID- 16038288 TI - Electrophoresis: the march of pennies, the march of dimes. AB - The present review encompasses ca. 65 years of history of developments in electrokinetic separations, taking as a starting point the year 1937, i.e. the official launching of Tiselius' moving boundary electrophoresis (MBE). The 1950s have been particularly rich in introducing novel methodologies in zone electrophoresis (ZE), thus bringing about the decline of MBE. Among them of extraordinary importance was the development of electrophoresis on agar gels coupled to immuno-diffusion at right angles, which brought a big revolution not only in biochemistry but also in clinical chemistry. Also the by now forgotten paper electrophoresis was a landmark in separation science, in that it implemented, in its "fingerprinting" version, the first genuine two-dimensional (2D) map, coupling orthogonally a charge to a hydrophobic scale separation, while permitting for the first time the detection of spot mutations, i.e. single amino acid replacements in a polypeptide chain, that paved the way to modern genetic analysis. Equally important was the introduction of starch-block electrophoresis, that brought about the notion of sieving and the first discontinuous buffers, refined, in the 1960s, by Ornstein and Davies with their classical papers combining multiphasic buffer systems to polyacrylamide gels, that went down to history as disc-electrophoresis. The 1960s also contributed with two fundamental techniques, isoelectric focusing (IEF) and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) that permitted to discriminate proteins solely on the basis of surface charge and molecular mass, respectively. The 1970s gave other fundamental contributions, such as isotachophoresis, the first example of a fully instrumental approach to electrophoresis, both in its analytical and preparative version (Tachophor and Tachofrac), 2D maps combining IEF to SDS-PAGE at right angles and silver staining techniques, that incremented sensitivity by 3 orders of magnitude. The 1980s generated immobilized pH gradients and capillary zone electrophoresis (CZE), two big players that dominated the electrokinetic horizon for all the 1990s and still in vigorous use in present days. The review terminates with a glimpse, in the third millennium, onto microchip technology and hyphenated techniques, notably direct interfacing of various electrophoretic separation methods with mass spectrometry (MS). PMID- 16038289 TI - Haplotyping by capillary electrophoresis. AB - The investigation of the genetic background and phenotype structures of complex diseases, such as cardiovascular or psychiatric disorders and tumors, is one of the most scrutinized fields of the post genomic era. Besides the multiplex analysis of genetic markers and polymorphisms throughout the whole genome, more and more attention is focused on the interaction between the etiological factors of these traits. Haplotype determination, rather than multiplex genotyping seems to be one of the first building blocks of this endeavor. This review focuses on the importance and theoretical background of haplotyping, and summarizes the recent examples of novel and emerging haplotyping techniques by capillary gel electrophoresis based DNA fragment analysis, a powerful tool for the examination of the inheritance of complex traits. PMID- 16038290 TI - Homogeneous gels for capillary electrochromatography. AB - Homogeneous gels represent a new type of (electro)chromatographic media possessing unique separation properties unmatched with any other chromatographic beds. It is important to emphasize that they principally differ from continuous beds, polymer rods (better known as monoliths), which are particulate separation media with pores permitting hydrodynamic flow through the columns. Monoliths, thus, are more similar to beds conventionally packed with beads, although the particles building up monolithic columns are usually smaller in size (few submicometers) and covalently linked together. Consequently, homogeneous gels deserve better the term "monoliths" having a non-particulate structure formed by crosslinked free polymer chains (according to a dictionary a monolith is a non modularized column). The goals of this minireview are to clarify the position of homogeneous gels among the separation media (including polymer solutions), to explain and to exemplify their outstanding (electro)chromatographic properties. This review gives hopefully a complete list of references to homogeneous gels developed for capillary electrochromatography. PMID- 16038291 TI - Harnessing dielectric forces for separations of cells, fine particles and macromolecules. AB - A review of conventional dielectrophoresis on a microchip platform is presented. The benefits of miniaturization, some device geometries used to accomplish on chip separations, and applications of these devices are discussed. PMID- 16038292 TI - Electrochemical properties of columns in capillary electrochromatography. I. Ohm's law, resistivity and field strength. AB - The most commonly used type of column in capillary electrochromatography (CEC) consists of a packed segment and an open (but buffer-filled) segment. The two segments differ importantly in two respects: firstly, their electrical resistivity; and secondly, their zeta potentials at a multitude of solid-liquid interfaces. Determination of the magnitude of these properties for each segment cannot be made using only results from the column as a whole. Instead, measurements of resistivity and zeta potentials of an entirely open, unpacked column can be used in conjunction with those of the CEC column to determine the electrochemical nature of both segments. This review of basic electrochemical properties will describe simple procedures that can be used to determine resistance, resistivity, conductivity, conductance, and field strength in each segment. In addition, it will be shown how the properties of each segment add together to give the same properties of the CEC column as a whole. The equations so derived will be applied to data from the literature and conclusions drawn from the results. PMID- 16038293 TI - Retrograde adsorption isotherms: an impossible fact? AB - "Retrograde" adsorption isotherms have been reported, but seem to violate thermodynamics and would cause concentration steps to migrate against the direction of fluid-phase flow. In general, what appears to be retrograde behavior is caused by one or more additional, uncontrolled variables. This is illustrated with adsorption of sulfonate on a weak-acid ion exchanger, where adsorption is accompanied by partial conversion of the ion exchanger to the sodium form. PMID- 16038294 TI - Exploiting pH mismatch in preparative high-performance liquid chromatographic recovery of ertapenem from mother liquor streams. AB - Preparative chromatography was successfully employed to recover ertapenem from mother liquor streams. The recovery process involved concentration of mother liquor stream by evaporation, purification by reversed-phase preparative high performance liquid chromatography (HPLC), and removal of chromatographic solvents in the recovered fractions by evaporation. HPLC feed was prepared by stripping off the organic solvents from the mother liquor using a wiped-film evaporator. Purification was first carried out on a 25 cm x 0.46 cm analytical column packed with 10-microm Kromasil C8 particles and then scaled up to a 25 cm x 5 cm preparative column. Gram-level recovery of ertapenem with high purity was achieved by exploiting a novel approach based on pH mismatch between the feed and the eluent. Purified ertapenem streams from preparative HPLC runs were combined, evaporated and recycled into the crystallizer for ertapenem isolation. PMID- 16038295 TI - Effect of charge regulation on steric mass-action equilibrium for the ion exchange adsorption of proteins. AB - A thermodynamic formalism is developed for incorporating the effects of charge regulation on the ion-exchange adsorption of proteins under mass-overloaded conditions as described by the steric mass-action (SMA) isotherm. To accomplish this, the pH titration behavior of a protein and the associated adsorption equilibrium of the various charged forms of a protein are incorporated into a model which also accounts for the steric hindrance of salt counterions caused by protein adsorption. For the case where the protein is dilute, the new model reduces to the protein adsorption model described recently by the authors which accounts for charge regulation. Similarly, the new model reduces to the steric mass-action isotherm developed by Brooks and Cramer which applies to mass overloaded conditions for the case where charge regulation is ignored so that the protein has a fixed charge. Calculations using the new model were found to agree with experimental data for the adsorption of bovine serum albumin (BSA) on an anion-exchange column packing when using reasonable physical properties. The new model was also used to develop an improved theoretical criterion for determining the conditions required for an adsorbed species to displace a protein in displacement chromatography when the pH is near the protein pI. PMID- 16038296 TI - Two-component protein adsorption kinetics in porous ion exchange media. AB - This work provides a theoretical analysis of multicomponent adsorption kinetics for conditions typical of protein adsorption in porous ion exchangers as well as experimental results for the adsorption of lysozyme/cytochrome c mixtures in the cation exchanger SP-Sepharose-FF. The theory predicts the formation of overshoots in the intraparticle concentration profiles and in the total amount adsorbed for the more weakly adsorbed component. An analytical solution valid for the case where the isotherms are rectangular is developed and found to be in good agreement with the limiting behavior of the general numerical solution of the model equations. The experimental results show that the two proteins are competitively adsorbed and that an overshoot of adsorbed cytochrome c occurs during simultaneous adsorption. Model predictions based on the assumption that the adsorption isotherms are rectangular and that lysozyme completely displaces cytochrome c are in qualitative and quantitative agreement with the experimental kinetics suggesting that the overshoot phenomena observed with multicomponent systems in these resins can be explained with a diffusion model without the need to account for flux coupling or electrophoretic contributions to transport. PMID- 16038297 TI - Competitive ion-exchange adsorption of proteins: competitive isotherms with controlled competitor concentration. AB - The competitive adsorption processes inevitably present in chromatographic separations of complex mixtures have not been extensively studied. This is partly due to the difficulty of measuring true competitive isotherms, in which all system parameters (including competitor concentrations) are held constant. We report a novel approach to determining competitive protein adsorption isotherms in which the competitor concentration is held constant across the entire isotherm. By using the heme prosthetic group in cytochrome b5 as a quantitative spectrophotometric label, competitive isotherms between cytochrome b5 and alpha lactalbumin can be constructed. Similarly, manganese-substituted protoporphyrin IX heme replacement allows the non-perturbing labeling of individual cytochrome b5 conservative surface charge mutants by replacement of a single atom in the interior of the protein. This labeling allows the study of competition between cytochrome b5 charge mutants of identical size and shape, which differ only in charge arrangement. Using these techniques, the effect of competing species on equilibrium behavior and the apparent heterogeneity of anion-exchange adsorbents in the presence of competitors can be quantitatively studied by fitting the data to two popular single-component binding models, the Temkin and the Langmuir Freundlich (L-F) isotherms. PMID- 16038298 TI - Molecular simulation study of the bonded-phase structure in reversed-phase liquid chromatography with neat aqueous solvent. AB - The dramatic loss of retention in reversed-phase liquid chromatography when switching to 100% aqueous solvent and stopping flow (depressurizing) has long intrigued separation scientists. Recent experimental evidence suggests that the observed loss of retention is due to the loss of pore wetting with subsequent loss of solvent penetration in the porous matrix. One of the prevalent explanations of this phenomenon has been that the bonded phase chains, typically octadecyl silane bound to porous silica, would undergo significant conformational changes, viz. collapse, under pure aqueous conditions. As a definitive means toward elucidating the conformation of bonded-phase chains under pure aqueous conditions, configurational-bias Monte Carlo simulations in the Gibbs ensemble were carried out for a system of dimethyl octadecyl silane of intermediate coverage bound to the (111) face of beta-cristobalite and immersed in pure water. The results of two sets of simulations, which were started from two entirely different starting configurations as a validity check toward reaching the same equilibrium distribution of states, show that chains are neither clustering together nor laying on the surface but rather have a broad distribution of orientations and of conformational states. The interface between the bonded and solvent phases is rough on a molecular level, and clusters of water molecules are sometimes found to adsorb at the silica surface. This computational study lends further evidence that the driving force for the loss of retention when switching to pure aqueous conditions and depressurizing is not the collapse of bonded-phase chains. PMID- 16038299 TI - Analyses of phenolic endocrine disrupting chemicals in marine samples by both gas and liquid chromatography-mass spectrometry. AB - With the renovation of Boston Harbor's Deer Island wastewater treatment plant and the extension of its diffuser pipes 15 km further into Massachusetts Bay, there arose the question whether the increased load of its secondary-treated wastewater contained significant amounts of phenolic endocrine disrupting chemicals (EDCs). Sampling from an oceanographic research vessel during the summers of 2003 and 2004 allowed for a unique opportunity to obtain clam, zooplankton, and bottom sediment samples. The samples were prepared by enhanced organic-solvent microwave digestion, followed by solid-phase extraction (SPE), derivatization and then analyzed by gas chromatography-mass spectrometry (GC-MS) or left un-derivatized and analyzed by LC-UV and liquid chromatography-mass spectrometry (LC-MS). The marine samples, especially parts of the clams, zooplankton and certain bottom sediments were found to contain primarily bisphenol A (BPA) at concentrations of 1-30 ng/g. PMID- 16038300 TI - Separation and characterization of underivatized oligosaccharides using liquid chromatography and liquid chromatography-electrospray ionization mass spectrometry. AB - Native cyclodextrin-based columns are particularly useful for the analysis of oligosaccharides because the retention of these carbohydrates is based mainly on the hydrogen bonding interactions of oligosaccharide hydroxyl groups with the stationary phase. Thus, the retention time predictably increases with the number of analyte hydroxyl groups, which corresponds to the elongation of the oligosaccharide chain. High-performance liquid chromatography (HPLC) coupled to electrospray ionization (ESI) mass spectrometry (MS) was used for the separation and characterization of underivatized oligosaccharide mixtures. With the limits of detection as low as 50 pg, all individual components of oligosaccharide mixtures (up to 11 glucose units long) were baseline resolved on a Cyclobond I 2000 column and detected using ESI-MS. Low flow rates and narrow I.D. columns increase the ESI-MS sensitivity significantly. The method showed potential usefulness for the sensitive and quick analysis of hydrolysis products of polysaccharides, and for trace levels of individual oligosaccharide or oligosaccharide isomers from biological systems. PMID- 16038301 TI - Theory of peak capacity in gradient elution. AB - Peak capacity is the best measure of the performance of a gradient separation. In this paper, the theory of peak capacity for the standard operating conditions of reversed-phase and ion-exchange chromatography is outlined. The influence of the operating conditions on the peak capacity of a separation are discussed. Finally, bandspreading phenomena in gradient chromatography are analyzed. PMID- 16038302 TI - Modeling of overloaded gradient elution of nociceptin/orphanin FQ in reversed phase liquid chromatography. AB - The Reversed-phase (RP) gradient elution chromatography of nociceptin/orphanin FQ (N/OFQ), a neuropeptide with many biological effects, has been modeled under linear and non-linear conditions. In order to do this, the chromatographic behavior has been studied under both linear and nonliner conditions under isocratic mode at different mobile phase compositions--ranging from 16 to 19% (v/v) acetonitrile (ACN) in aqueous trifluoracetic acid (TFA) 0.1% (v/v)-on a C-8 column. Although the range of mobile phase compositions investigated was quite narrow, the retention factor of this relatively small polypeptide (N/OFQ is a heptadecapeptide) has been found to change by more than 400%. In these conditions, gradient operation resulted thus to be the optimum approach for non linear elution. As the available amount of N/OFQ was extremely reduced (only a few milligrams), the adsorption isotherms of the peptide, at the different mobile phase compositions examined, have been measured through the so-called inverse method (IM) on a 5 cm long column. The adsorption data at different mobile phase compositions have been fitted to several models of adsorption. The dependence of the isotherm parameters on the mobile phase composition was modeled by using the linear solvent strength (LSS) model and a generalized Langmuir isotherm that includes the mobile phase composition dependence. The overloaded gradient separation of N/OFQ has been modeled by numerically solving the equilibrium dispersive (ED) model of chromatography under a selected gradient elution mode, on the basis of the previously determined generalized Langmuir isotherm. The agreement between theoretical calculations and experimental overloaded band profiles appeared reasonably accurate. PMID- 16038303 TI - Binding behaviour and conformational properties of globular proteins in the presence of immobilised non-polar ligands used in reversed-phase liquid chromatography. AB - The thermodynamic and extra-thermodynamic dependencies of five types of cytochrome c in water-acetonitrile mixtures of different composition in the presence of immobilised n-octyl ligands as a function of temperature from 278 K to 338 K have been investigated. The corresponding enthalpic, entropic and heat capacity parameters, deltaHdegrees assoc, deltaS degrees assoc and delta C degrees p, have been evaluated from the observed non-linear Van't Hoff plots of these globular proteins in these heterogeneous systems. The relationships between the free energy dependencies, various molecular parameters and extra thermodynamic dependencies (empirical correlations) of these protein-non-polar ligand interactions have also been examined. Thus, the involvement of enthalpy entropy compensation effects has been documented for the binding of these cytochrome cs to solvated n-octyl ligands. Moreover, the results confirm that this experimental approach permits changes in molecular surface area due to the unfolding of these proteins on association with non-polar ligands as a function of temperature to be correlated with other biophysical properties. This study thus provides a general procedure whereby the corresponding free energy dependencies of globular proteins on association with solvated non-polar ligands in heterogeneous two-phase systems can be quantitatively evaluated in terms of fundamental molecular parameters. PMID- 16038304 TI - Solid-phase capture and release of arginine peptides by selective tagging and boronate affinity chromatography. AB - A method for the selection of arginine-containing peptides from a mixture by a solid phase capture and release technique is presented. The method is based on the covalent modification of the guanidine group of arginine with 2,3-butanedione and phenylboronic acid under alkaline conditions. Using polymeric materials with immobilised phenylboronic acid the arginine-peptides can be captured on a solid support while arginine-free peptides are not covalently bound and can be washed away. Finally, the arginine-peptides can be cleaved again from the boronic acid beads due to the reversibility of the reaction. The recovered peptides are then analysed by liquid chromatography-tandem mass spectrometry. The method was optimised with model peptides with regard to the non-specific binding of arginine free peptides and quantitative cleavage of the label after the selection step. Using an adequate protocol, the applicability towards more complex samples was successfully tested with a tryptic digest of a mixture of three standard proteins. PMID- 16038305 TI - Characterisation and evaluation of metal-loaded iminodiacetic acid-silica of different porosity for the selective enrichment of phosphopeptides. AB - Silica particles of different porosity were functionalised with iminodiacetic acid (IDA) and loaded with Fe(III) to yield immobilised metal affinity chromatography stationary phases (Fe(III)-IDA-silica) for phosphopeptide enrichment. The elution step of bound phosphopeptides was optimised with a 32P radioactive labelled peptide by a comprehensive study. Several elution systems, including phosphate buffers of different pH and concentration and ethylenediaminetetraacetic acid solutions were employed. Furthermore the effect of support porosity on elution behaviour was investigated. Under best conditions recoveries higher than 90% were achieved. A solid-phase extraction (SPE) protocol was developed for fractionation of phosphorylated and non-phosphorylated peptides and desalting of the fractions which is essential for subsequent mass spectrometric analysis by the combination of Fe(III)-IDA-silica and C18-silica particles. The pH of the loading buffer was found to be a critical parameter for the efficiency of the SPE protocol. As tryptic digests of alpha-lactalbumin, lysozyme and ribonuclease A mixed with three synthetic phosphopeptides were fractionated, pH 2.5 provided minimal proportion of unspecific bound peptides when comparing the fractions after mu-LC-electrospray ionization MS separation. The effect of a sample derivatisation reaction (methylation) on the efficiency of phosphopeptide enrichment was further investigated. Blocking carboxylate groups by methyl ester formation totally prevented unspecific interaction with the immobilised Fe(III) ions, but generated partially methylated phosphopeptides that increased the complexity of the phosphorylated fraction. PMID- 16038306 TI - Detection of Nepsilon-monomethyllysine using high-performance liquid chromatography and high-performance liquid chromatography-mass spectrometry. AB - Nepsilon-Monomethyllysine was identified in the serum, urine, brain, and liver samples of rats treated per os with L-deprenyl. The identification procedure included reaction with Fmoc chloride, clean-up, and analysis using HPLC-UV-MS. Oral administration of (-)-N-14C-methyl-N-propynyl(2-phenyl-1 methyl)ethylammonium hydrochloride L-deprenyl) to rats resulted in transfer of the radiolabelled methyl group to the Nepsilon-amino group of the endogenous lysine. The radiolabelled Nepsilon-monomethyllysine was urinary eliminated together with the other radiolabelled deprenyl metabolites, such as deprenyl-N oxide and methamphetamine. The presence of Nepsilon-monomethyllysine has also been traced, and its concentrations were compared in the serum, liver and brain of rats subjected to L-deprenyl treatment. Methyl group transfer from the L deprenyl to endogenous compounds; and the urinary elimination of their products may offer a vital way to eliminate or to decrease the degree of drug transmethylation to the lysine constituents of blood vessels' proteins. PMID- 16038307 TI - High temperature fast chromatography of proteins using a silica-based stationary phase with greatly enhanced low pH stability. AB - Reversed-phase liquid chromatography (RPLC) is very widely used for the separation and characterization of proteins and peptides. A novel type of highly stable silica-based stationary phase has been developed for protein separations. A dense monolayer of dimethyl-(chloromethyl)phenylethyl)-chlorosilane (DM-CMPES) on the surface of silica is "hyper-crosslinked" with a polyfunctional aromatic crosslinker through Friedel-Crafts chemistry resulting in stationary phases with extraordinary stability in acidic media. Elemental analysis data confirm the high degree of cross-linking among the surface groups. The hyper-crosslinked phases are extremely stable under highly acidic mobile phase conditions even at a temperature as high as 150 degrees C. A wide-pore (300 A) material made in this way is used here to separate proteins by a reversed-phase mechanism and compared to a commercially available "sterically protected" C18 phase. For small molecules, including neutral and basic compounds, these crosslinked phases give comparable peak shape and efficiency to the commercial phase. Our results show that no pore blockage takes place as commonly afflicts polymer coated phases. In consequence, protein separations on the new phases are acceptable. Using strong ion-pairing reagents, such as HPF6, improves the separation efficiency. Compared to the commercial phases, these new phases can be used at lower pHs and much higher temperatures thereby enabling much faster separations which is the primary focus of this work. Better efficiency for proteins was obtained at high temperature. However, at conventional linear velocities the instability of proteins at high temperature becomes a problem which establishes an upper temperature limit. Uses of a narrowbore column and high flow rates both solves this problem by reducing the time that proteins spend on the hot column and, of course, speeds up the separation of the protein mixture. Finally, an ultrafast gradient (<1 min) protein separation was obtained by utilizing the high temperature and thus high linear velocities afforded by the extreme stability of these new phases. The phases are stable even after 50h of exposure to 0.1% TFA at 120 degrees C. This paper is dedicated to the memory of Csaba Horvath whose work in high temperature HPLC inspired the development of the stationary phases described here. PMID- 16038308 TI - Hydrophobic interaction chromatography of proteins. III. Unfolding of proteins upon adsorption. AB - Hydrophobic interaction chromatography (HIC) exploits the hydrophobic properties of protein surfaces for separation and purification by performing interactions with chromatographic sorbents of hydrophobic nature. In contrast to reversed phase chromatography, this methodology is less detrimental to the protein and is therefore more commonly used in industrial scale as well as in bench scale when the conformational integrity of the protein is important. Hydrophobic interactions are promoted by salt and thus proteins are retained in presence of a cosmotropic salt. When proteins are injected on HIC columns with increasing salt concentrations under isocratic conditions only, a fraction of the applied amount is eluted. The higher the salt concentration, the lower is the amount of eluted protein. The rest can be desorbed with a buffer of low salt concentration or water. It has been proposed that the stronger retained protein fraction has partially changed the conformation upon adsorption. This has been also corroborated by physicochemical measurements. The retention data of 5 different model proteins and 10 different stationary phases were evaluated. Partial unfolding of proteins upon adsorption on surfaces of HIC media were assumed and a model describing the adsorption of native and partial unfolded fraction was developed. Furthermore, we hypothesize that the surface acts as catalyst for partial unfolding, since the fraction of partial unfolded protein is increasing with length of the alkyl chain. PMID- 16038309 TI - Effect of pH changes on water release values in hydrophobic interaction chromatographic systems. AB - The effect on pH on protein binding in HIC systems was investigated. Isocratic experiments were carried out to determine the capacity factors of various proteins as a function of temperature, pH and salt type. This paper presents a framework based on the Maxwell linkage function for estimating the number of released water molecules during the adsorption/desorption process due to a change of buffer pH. This approach also enables one to predict the effect of pH change on the water released values upon binding at any temperature condition. The results indicate that the total number of released water molecules (delta nu) for a pH change increased more on aromatic surfaces (phenyl Sepharose) than on aliphatic resins (butyl Sepharose). The results also indicate that the total number of released water molecules (deltanu) for a pH change increased with salt concentration and when changing from chaotropic to kosmotropic salts. The (deltanu) values also increased as the buffer pH approached the protein's pI, and decreased away from its pI. This work helps to establish a framework for the investigation of pH effects on protein selectivity in HIC systems. PMID- 16038310 TI - Affinity chromatography with monolithic capillary columns. II. Polymethacrylate monoliths with immobilized lectins for the separation of glycoconjugates by nano liquid affinity chromatography. AB - Monolithic capillary columns with surface bound lectin affinity ligands were introduced for performing lectin affinity chromatography (LAC) by nano-liquid chromatography (nano-LC). Two kinds of polymethacrylate monoliths were prepared, namely poly(glycidyl methacrylateco-ethylene dimethacrylate) and poly(glycidyl methacrylate-co-ethylene dimethacrylate-co-[2-(methacryloyloxy)ethyl]trimethyl ammonium chloride) to yield neutral and cationic macroporous polymer, respectively. Two lectins including concanavalin (Con A) and wheat germ agglutinin (WGA) were immobilized onto the monolithic capillary columns. The neutral monoliths with immobilized lectins exhibited lower permeability under pressure driven flow than the cationic monoliths indicating that the latter had wider flow-through pores than the former. Both types of monoliths with immobilized lectins exhibited strong affinity toward particular glycoproteins and their oligosaccharide chains (i.e., glycans) having sugar sequences recognizable by the lectin. Due to the strong binding affinity, the monoliths with surface bound lectins allowed the injection of relatively large volume (i.e., several column volumes) of dilute samples of glycoproteins and glycans thus allowing the concentration of the glycoconjugates and their subsequent isolation and detection at low levels (approximately 10(-8) M). To further exploit the lectin monoliths in the isolation of glycoconjugates, two-dimensional separation schemes involving LAC in the first dimension and reversed-phase nano-LC in the second dimension were introduced. The various interrelated methods established in this investigation are expected to play a major role in advancing the sciences of "nano-glycomics". PMID- 16038311 TI - Nitromethane as solvent in capillary electrophoresis. AB - Nitromethane has several properties that make it an interesting solvent for capillary electrophoresis especially for lipophilic analytes that are not sufficiently soluble in water: freezing and boiling points are suitable for laboratory conditions, low viscosity leads to favourable electrophoretic mobilities, or an intermediate dielectric constant enables dissolution of electrolytes. In the present work we investigate the change of electrophoretically relevant analyte properties - mobilities and pKa values - in nitromethane in dependence on the most important experimental conditions determined by the background electrolyte: the ionic strength, I, and the pH. It was found that the mobility decreases with increasing ionic strength (by, e.g. up to 30% from I = 0 to 50 mmol/L) according to theory. An appropriate pH scale is established by the aid of applying different concentration ratios of a buffer acid with known pKa and its conjugate base. The mobility of the anionic analytes (from weak neutral acids) depends on the pH with the typical sigmoidal curve in accordance with theory. The pKa of neutral acids derived from these curves is shifted by as much as 14 pK units in nitromethane compared to water. Both findings confirm the agreement of the electrophoretic behaviour of the analytes with theories of electrolyte solutions. Separation of several neutral analytes was demonstrated upon formation of charged complexes due to heteroconjugation with chloride as ionic constituent of the background electrolyte. PMID- 16038312 TI - Rapid quantitative capillary zone electrophoresis method for monitoring the micro heterogeneity of an intact recombinant glycoprotein. AB - A simple high-resolution capillary zone electrophoresis (CZE) method capable of rapidly assessing the micro-heterogeneity of a 24 kDa molecular weight glycoprotein, has been developed. Separation is carried out using a bare silica capillary at a pH of 2.5 in a commercially available electrophoresis buffer system composed of triethanolamine and phosphoric acid. Over 30 peaks were detected within a run time of 15 min using a 27 cm capillary and approximately 60 peaks were detected using a 77 cm capillary. Although most of the peaks arise from differences in the oligosaccharide structures present on the one glycosylation site on this molecule, other forms of micro-heterogeneity due to the presence of the nonglycosylated form of this glycoprotein and various types of chemical degradation, e.g., deamidation, are also responsible for the multitude of peaks observed. Although the exact chemical identity of each peak in the resulting electropherogram of this glycoprotein is not known, useful information can be obtained for assessing comparability, stability, and batch consistency. Factors impacting the resolution, precision, accuracy, and robustness of the assay are also discussed along with inherent advantages and limitations associated with measuring the micro-heterogeneity of intact glycoproteins. PMID- 16038313 TI - Dynamic pH junction technique for on-line preconcentration of peptides in capillary electrophoresis. AB - A method based on the presence of a dynamic pH junction within the capillary to induce band narrowing for enhanced detection sensitivity for some peptides is presented. This technique is predicated on a sharp reduction in an analyte's migration velocity following a reversal of its electrophoretic direction from the acidic sample zone to the basic BGS zone. Larger-than-usual injection volumes of samples in relatively high-conductivity matrices were enabled, without degrading peak shape, resolution and efficiency. The size of the original sample plug was reduced by as much as 38-fold, and improvement in detector response in terms of peak height by as much as 124-fold was obtained. The effects of pH and concentration of the sample matrix, and the length of sample injection on the efficiency of the technique are discussed. PMID- 16038314 TI - Separation, detection, and identification of peptides by ion-pair reversed-phase high-performance liquid chromatography-electrospray ionization mass spectrometry at high and low pH. AB - Bioactive peptides and tryptic digests of various proteins were separated under acidic and alkaline conditions by ion-pair-reversed-phase high-performance liquid chromatography (RP-HPIPC) in 200 microm I.D. monolithic, poly(styrene divinylbenzene)-based capillary columns using gradients of acetonitrile in 0.050% aqueous trifluoroacetic acid, pH 2.1, or 1.0% triethylamine-acetic acid, pH 10.6. Chromatographic performances with mobile phases of low and high-pH were practically equivalent and facilitated the separation of more than 50 tryptic peptides of bovine serum albumin within 15-20 min with peak widths at half height between 4 and 10 s. Neither a significant change in retentivity nor efficiency of the monolithic column was observed during 17-day operation at pH 10.6 and 50 degrees C. Upon separation by RP-HPIPC at high-pH, peptide detectabilities in full-scan negative-ion electrospray ionization mass spectrometry (negESI-MS) were about two to three times lower as compared to RP-HPIPC at low-pH with posESI-MS detection. Tandem mass spectra obtained by fragmentation of deprotonated peptide ions in negative ion mode yielded interpretable sequence information only in a few cases of relatively short peptides. However, in order to obtain sequence information for peptides separated with alkaline mobile phases, tandem mass spectrometry (MS/MS) could be performed in positive ion mode. The chromatographic selectivities were significantly different in separations performed with acidic and alkaline eluents, which facilitated the fractionation of a complex peptide mixture obtained by the tryptic digestion of 10 proteins utilizing off-line, two dimensional RP-HPIPC at high pH x RP-HPIPC at low pH and subsequent on-line identification by posESI-MS/MS. PMID- 16038315 TI - Evaluation of multidimensional (ion-exchange/reversed-phase) protein separations using linear and step gradients in the first dimension. AB - The performance characteristics of multidimensional liquid chromatographic protein separations were evaluated using on-line electrospray mass detection, and a novel workflow for automated LC/MS data processing. Two-dimensional ion exchange/reversed-phase LC separations of Escherichia coli cytosol were conducted using either a continuous linear or discontinuous step gradient in the first dimension. Chromatographic profiles of the top 100 most abundant components were characterized to assess overall separation reproducibility within each mode, and to characterize differences in component distribution between the two modes of operation. Analysis of the resulting data indicates that multidimensional separations of complex protein mixtures can be done reproducibly. Furthermore, under the conditions employed within this study, a linear first dimension gradient was more effective at fractionating the protein mixture, distributing fewer major components to multiple second dimension cycles than an equivalent step gradient. The application of on line mass spectrometry, and automated processing of the resulting data, proved valuable for producing component level analysis of multidimensional protein separations. PMID- 16038316 TI - Use of electrokinetic measurements for characterization of columns used in capillary electrochromatography. AB - The separation mechanism in capillary electrochromatography (CEC) is a hybrid differential migration process, which entails the features of both high performance liquid chromatography (HPLC) and capillary zone electrophoresis (CZE), i.e., chromatographic retention and electrophoretic migration. The focus of this paper is on the use of electrokinetic data, such as current, electroosmotic flow (EOF) and column efficiency measurements, that are readily available, for an improved understanding of CEC separations. A framework is presented here for the use of this data for evaluation of a variety of performance parameters including, conductivity ratio, interstitial EOF mobility, porosity, and zeta potential. This framework is applied for characterization of two monolithic columns with different chemistry that were manufactured in-house. The above-mentioned performance parameters were calculated for the two columns and it is found that the poly(VBC-EGDMA-SWNT) monolithic column with the GPTMS PEI coating offers a significantly improved flow distribution in comparison to the poly(VBC-EGDMA) monolithic column. This observation is confirmed by performing separation of peptides on the two columns and height equivalent of a theoretical plate (HETP) measurements on the resulting peaks. It is shown that following our approach leads to an improved understanding of the separations achieved with the columns and to better column design. PMID- 16038317 TI - PH effects on drug interactions with lipid bilayers by liposome electrokinetic chromatography. AB - Liposome electrokinetic chromatography (LEKC) provides convenient and rapid methods for studying drug interactions with lipid bilayers using liposomes as a pseudostationary phase. LEKC was used to determine the effects of pH on the partitioning of basic drugs into liposomes composed of zwitterionic phosphatidylcholine (PC), anionic phosphatidylglycerol (PG), and cholesterol, which mimic the composition of natural cell membranes. An increase in pH results in a smaller degree of ionization of the basic drugs and consequently leads to a lower degree of interaction with the negatively charged membranes. From the LEKC retention data, the fractions of drugs distributed in the bulk aqueous and the liposome phase were determined at various pH values. Finally, lipid mediated shifts in the ionization constants of drugs were examined. PMID- 16038318 TI - Characterization of flow and voltage profiles in planar electrochromatography. AB - Planar electrochromatography (PEC) is a new technology for thin layer chromatography (TLC) where the separation is driven by electroosmotic forces, not capillary action. This allows for much faster and more efficient chromatography in a planar format. Care needs to be taken when performing these experiments because voltage and flow characteristics can change through a single run, due to buffer gradients, temperature changes (Joule heating) and localized plate heterogeneity. We have designed a PEC instrument and cover grid to allow investigation of flow and voltage characteristics as solvent moves across a TLC plate. Our unique cover grid allows monitoring voltage at eight discrete points between the positive and negative reservoirs. A linear relationship between voltage and distance should be seen, giving a constant voltage drop across a plate, but this did not occur. This non-linear function changes over time, following the plate equilibration. Once a plate is equilibrated, voltage and flow characteristics remain fairly constant. Theoretical calculations support the physical observations. Larger plate widths (5 cm) were also briefly investigated and it is concluded that large width plates could be easily implemented to maintain multiple sample capability. PMID- 16038319 TI - Use of mixed-mode sorbents for the electrochromatographic separation of thrombin receptor antagonistic peptides. AB - In this study, the thrombin receptor antagonistic peptide TRAP-1 and its alanine scan analogues, TRAP 2-6, have been employed as probes to characterise the performance of C18/SCX mixed-mode capillary electrochromatographic (CEC) columns. It was found that the resolution of this group of peptides could only be achieved in a narrow pH range with phosphate-based running electrolytes. The influence of the running electrolyte composition, e.g. the buffer choice, the ionic strength, the pH and the organic solvent content, on the electroosmotic flow (EOF) of these mixed-mode CEC columns was investigated. In addition, the retention mechanism for this group of peptide probes in the electrochromatographic process was studied by examining the effect of varying the running electrolyte composition. As a result, it can be concluded that the electrochromatographic separation of this set of peptides was mediated by a combination of electrophoretic migration and chromatographic retention involving both hydrophobic as well as ion exchange interactions. By modulating the running electrolyte composition, the hydrophobic or ion exchange components of the interaction process could be made to dominate the chromatographic retention of the peptides. Based on this strategy, a high resolution separation of six closely related synthetic peptides was demonstrated with this mixed-mode CEC system. PMID- 16038320 TI - Amino acids determination using capillary electrophoresis with on-capillary derivatization and laser-induced fluorescence detection. AB - Free amino acids have been derivatized on-capillary with 3-(2-furoyl)quinoline-2 carboxaldehyde (FQ) and analyzed using a laboratory-made capillary electrophoresis apparatus with laser-induced fluorescence detection. Several parameters that control on-capillary derivatization of amino acids, including pH, mixing time, reaction time, concentration of the derivatization reagents (potassium cyanide and FQ) and solvent of FQ, as well as the temperature of mixing and reaction were optimized. Repeatabilities better than 1.8% for migration time and 7.8% for peak height were obtained. Assay detection limits for the different amino acids ranged from 23 nM for glycine to 50 nM for lysine and glutamic acid. The methods developed were applied to the analysis of several amino acids in pharmaceutical preparations and plasma samples. Results showed a good agreement with those obtained using an amino acid autoanalyzer for the same samples. PMID- 16038321 TI - Polypyrrole-coated capillaries for capillary zone electrophoresis. AB - Fused-silica capillaries are permanently coated by silanization with 3-{[3-(N pyrrole)-2-hydroxypropyl]amino}propyltriethoxysilane followed by oxidative polymerization of the pyrrole moieties with iron (III) or peroxodisulfate in the presence of chloride, perchlorate, or dextransulfate as anions. This approach allows to modulate the EOF in its magnitude as well as in its direction. With the small anions chloride and perchlorate, the EOF is reversed below pH 5 while with the large dextransulfate polyanions (DS) the EOF is relatively constant over the pH range from 2.5 to 9.4. This can be of advantage at low pH, at which the EOF of uncoated capillaries is close to zero. Application for separation of some herbicides is shown. The lifetime of PP-modified capillaries is satisfactory: the decrease in EOF is less than 3% during 80 analyses (160 min) and less than 5% over three months of storage. The reproducibility of capillary modification is about 5% (RSD of EOF). PMID- 16038322 TI - Application of main component fraction collection method for purification of compound libraries. AB - In order to support high-throughput library purification, a novel UV triggered fraction collection method was developed in which a maximum-seeking-algorithm driven, six-port valve collects the largest chromatographic peak. This straightforward strategy achieves the one sample-one fraction approach, thus resulting in a simpler and less error prone workup procedure. The effectiveness of this main component fraction collection method will be illustrated here by the results of the purification of compound libraries (altogether 6086 compounds, having an averaged success rate of 79.4%). Advanced applications, where the desired component differs from the main component, will also be discussed. PMID- 16038323 TI - Application of capillary zone electrophoresis and reversed-phase high-performance liquid chromatography in the biopharmaceutical industry for the quantitative analysis of the monosaccharides released from a highly glycosylated therapeutic protein. AB - Two assays for the quantitative determination of the neutral and amino monosaccharides attached to a therapeutic glycoprotein were developed using capillary zone electrophoresis (CZE) and RP-HPLC. These assays meet the strict batch release requirements of the quality control in biopharmaceutical industry. The monosaccharides were released from the glycoprotein by hydrolysis with 2N trifluoroacetic acid. In the CZE assay the monosaccharides were reacetylated prior to derivatization with 8-aminopyrenesulfonic acid (APTS), reacetylation in the glycoprotein matrix was investigated in detail. The RP-HPLC method used pre column derivatization with anthranilic acid in methanol-acetate-borate reaction medium; reacetylation was not necessary. However, epimerization of the different monosaccharides was observed and studied in detail. For the quantitative assay, separation of the amino-monosaccharide epimers had to be developed. The HPLC assay was validated. PMID- 16038324 TI - Comparison of silanization/hydrosilation and organosilanization modification procedures on etched capillaries for electrokinetic chromatography. AB - Etched capillaries for use in open tubular electrochromatography are modified by silanization/hydrosilation and organosilanization. The migration behavior of both types of capillaries is evaluated with small basic molecules, peptides and proteins. Comparisons of peak symmetry and efficiency are used to measure the effectiveness of the two methods for modifying the etched surface. From this information, the suitability of each method for use with etched capillaries can be determined. PMID- 16038325 TI - Buffer system for the separation of neutral and charged small molecules using micellar electrokinetic chromatography with mass spectrometric detection. AB - An organic buffer system will be discussed that is suitable for the separation of neutral as well as charged molecules be means of micellar electrokinetic chromatography (MEKC). The buffers are based on the combination of a long chain alkyl acid, such as lauric acid with ammonium hydroxide or an organic base such as tris-hydroxymethylaminomethane (Tris). The resulting buffer system is able to separate neutral compounds based on its micellar properties. These buffers exhibit much reduced conductivity compared to traditional MEKC buffers, such as sodium dodecylsulfate (SDS), which contain inorganic salts. They also have inherent buffer capacity at high pH resulting from the basic buffer component, which in our studies had pK values from about 8-11. The separations that were observed showed high efficiency with plate counts in many cases above 500,000 plates per meter. The reduced conductivity allowed for the application of much higher electric fields, resulting in very fast analysis times. Alternatively, an increase in detection sensitivity could be achieved, as the reduced conductivity allowed for the use of capillaries with lager internal diameters. Combinations of different alkyl acids and organic bases provided for significant flexibility in selectivity tuning. Finally, the fact that the organic micellar buffer systems discussed here do not contain inorganic ions, allows for coupling with mass spectrometric (MS) detection. The possibility of MS detection combined with the high speed in analysis that can be obtained using these organic buffer systems, could make this approach an interesting option for high throughput analysis of combinatorial libraries. PMID- 16038326 TI - Preparation and evaluation of poly(polyethylene glycol methyl ether acrylate-co polyethylene glycol diacrylate) monolith for protein analysis. AB - A poly(polyethylene glycol methyl ether acrylate-co-polyethylene glycol diacrylate) monolith was prepared by UV-initiated polymerization. Methanol and ethyl ether were selected as porogens from a variety of organic solvents to achieve the desirable characteristics of the monolith. The preparation of the monolith could be achieved within 10 min. The monolith was macroscopically homogeneous, had low flow resistance, and did not swell or shrink significantly in tetrahydrofuran. Inverse size-exclusion data indicate that the monolith had a total porosity of 75.4% and an internal porosity of 9.1%. The monolith could be used for size-exclusion separation of peptides, although it could not separate proteins with molecular masses between 10 and 100 K due to its unique pore size distribution. It was found to resist adsorption of proteins in capillary liquid chromatography when using 100 mM phosphate buffer (pH 7.0) containing 0.5 M NaCl. Complete recovery of both acidic and basic proteins was achieved. The monolith can be used for applications in which inert materials are required for protein analysis. PMID- 16038327 TI - Metal oxide monolithic columns. AB - Metal oxide monoliths composed of ZrO2 and HfO2 have been synthesized in situ inside capillary columns. The material shows globular-like structure and through pores. Capillary electrochromatography and capillary liquid chromatography were performed in a monolithic column with the HfO2 material. Separation of a simple sample mixture showed the potential of the new metal oxide monolithic columns. PMID- 16038328 TI - Studies on the use of needle-free injection device on proteins. AB - In the following communication we report the evaluation of 18 proteins that were processed by a specific needle free injection device. The processed protein samples were analyzed by two HPLC techniques, reversed-phase liquid chromatography (RPLC) and size-exclusion chromatography (SEC). These techniques are two of the most widely used analytical techniques in the biopharmaceutical industry for the characterization, integrity assessment and stability study of peptide and protein products. The results indicate that needle free injection, using the specific device of this study, is not damaging to the studied proteins and does not generate aggregates. We found no evidence of the predicted possible effects of needle free injections, and concluded that needle free delivery is in general not different than any other delivery system and that its use should be evaluated on a case by case basis. It has to be noted that there are various needle free device designs and our work was performed using an Iject from Bioject. Our conclusions therefore should be limited to the Iject design we used in this study. In the reported experiments we used commercially available (economical) model proteins, which facilitate the use of the results for future comparison and reference. The work reported here can serve as a reference to illustrate the benign nature of our needle free injection device. It also highlights an interesting analogy between a set of phobias that were seen to have plagued the early stages of biochemistry and HPLC, on the one hand, and some attitudes that appear to hinder the widespread acceptance of needle free injection at present time, on the other. These phobias were identified and named by Professor Csaba Horvath, the father of HPLC, as barophobia, siderophobia and lithophobia. Today a wealth of evidence is available to indicate that those phobias are ungrounded and that the negative observations can be explained in most cases by adsorption and prevented by proper formulations and solvent conditions. PMID- 16038329 TI - Automated headspace-solid-phase micro extraction-retention time locked-isotope dilution gas chromatography-mass spectrometry for the analysis of organotin compounds in water and sediment samples. AB - An automated method for the simultaneous determination of six important organotin compounds namely monobutyltin (MBT), dibutyltin (DBT), tributyltin (TBT), monophenyltin (MPhT), diphenyltin (DPhT) and triphenyltin (TPhT) in water and sediment samples is described. The method is based on derivatization with sodium tetraethylborate followed by automated headspace-solid-phase micro extraction (SPME) combined with GC-MS under retention time locked (RTL) conditions. Home synthesized deuterated organotin analogues were used as internal standards. Two high abundant fragment ions corresponding to the main tin isotopes Sn118 and Sn120 were chosen; one for quantification and one as qualifier ion. The method was validated and excellent figures of merit were obtained. Limits of quantification (LOQs) are from 1.3 to 15 ng l(-1) (ppt) for water samples and from 1.0 to 6.3 microg kg(-1) (ppb) for sediment samples. Accuracy for sediment samples was tested on spiked real-life sediment samples and on a reference PACS-2 marine harbor sediment. The developed method was used in a case-study at the harbor of Antwerp where sediment samples in different areas were taken and subsequently screened for TBT contamination. Concentrations ranged from 15 microg kg(-1) in the port of Antwerp up to 43 mg kg(-1) near a ship repair unit. PMID- 16038330 TI - [Non-alcoholic fatty liver--life of overindulgence with fatty liver]. PMID- 16038331 TI - [Therapy refractory hypertension: mask helped where drugs failed]. PMID- 16038332 TI - [After large studies by federal health insurance: what is the future of acupuncture?]. PMID- 16038333 TI - [Can natural substances prevent or improve type 2 diabetes mellitus? Coffee and cinnamon for control of hyperglycemia]. PMID- 16038334 TI - [In type 2 diabetic patients look for silent myocardial ischemia. Yes, but only after preselection]. PMID- 16038335 TI - [Vitamin D deficiency promotes tumor development. Sun bathing for cancer prevention? (interview by Dr. Thomas Meissner)]. PMID- 16038336 TI - [Knee as vulnerable joint--inappropriate training, too much excess weight, an old injury?]. PMID- 16038337 TI - [Chronic knee joint discomfort]. AB - Chronic pain in the knee joint is frequently a sign of arthrosis in adults. This must be clearly differentiated from other knee problems. Patellofemoral stress syndrome (occurs mostly in young people) and arthritis with effusion in the knee joint after long and mostly unusual stress also allow only a reduced function of the knee joint. However, even when the knee joint is still fully functional, chronic problems could already exist: For example, for joggers, iliotibial band friction syndrome (runner's knee) or after high unphysiological stress, patellar tendinopathy (jumper's knee). These must be differentiated from pes anserinus syndrome and a plica mediopatellaris. PMID- 16038338 TI - [Hip pain--a problem of differential diagnosis in the primary care practice]. AB - The criterion "positive or negative capsular pattern" serves as a signpost in the differential diagnosis of pain in the groin. Slipped capital femoral epiphysis, Legg-Calve-Perthes disease, necrosis of the femoral head in adults, and the coxarthrosis of athletes belong to the first group. Inguinal hernia, osteitis pubis, and adductor tendinosis have no capsular pattern. Pain not localized in the groin is referred to as pelvic girdle pain. Pelvic girdle pain can arise suddenly after an intense trauma such as after a traffic accident, overstraining during sports or as apophyseal injuries. If chronic pelvic girdle pain exists, the cause of the problem is frequently primary or insertion tendinosis or bursitis. PMID- 16038339 TI - [Which cardiac patients may be allowed to travel by air?]. AB - During airline flights, cardiological patients in particular are put at stress by the decrease in pressure in the passenger cabin and the associated decrease in oxygen partial pressure in the blood. Possible risks are, in particular, rhythm disorders, myocardial ischemic states or an excessive increase in pulmonary artery pressure. Flight travel may be permitted at the earliest two weeks after a myocardial infarction or coronary event. It should be forbidden in patients with decompensated cardiac failure, instable angina pectoris, severe pulmonary arterial or arterial hypertension, uncontrolled arrhythmias and surgery on the heart within a period of 2 weeks prior to the intended flight. PMID- 16038340 TI - [Chronic pain due to degenerative and inflammatory musculoskeletal diseases in the elderly. Observational study investigating dosage, course of treatment, effectiveness and tolerability of tilidine/naloxone]. PMID- 16038341 TI - [Unusual obstruction of the small intestine]. PMID- 16038342 TI - [10 minute consultation: unexpected weight loss as the main symptom. "At the beginning I was still happy that the kilos dropped"]. PMID- 16038344 TI - [French health cooperation with developing countries]. PMID- 16038343 TI - [Diagnosis at a glance. Young man risks more than a fat lip]. PMID- 16038345 TI - [Contribution of the French Armed Forces Health Services to relief in Indonesia following the tsunami on December 26, 2004]. AB - As of early January 2005, the French Armed Forces Health Services were on the ground in Indonesia to contribute to relief operations following the tsunami in Indonesia on December 26, 2004. Findings of preliminary investigation performed upon deployment of the first French elements demonstrated that a major priority was to speed up vaccination against measles in children between the ages of 6 months and 15 years. A vaccination campaign was planned in cooperation with Indonesian authorities and UNICEF. Implementation involved training of personnel and setting up vaccination clinics in refugee camps, schools, and neighborhoods in the city of Meulaboh. The vaccination campaign was hampered by a number of problems in particular involving logistics and required constant supervision. A survey to evaluate the results of the campaign in terms of vaccination coverage showed that the objective of vaccinating 70% of children in refugee camps was achieved. The survey provided other child health indicators that will be helpful to local authorities taking over after the departure of the French Armed Forces Health Services. PMID- 16038346 TI - [Experience of Handicap International in providing humanitarian relief in region near Aceh, Indonesia from March 1 to 27, 2005]. AB - This report describes the experience of the author in March 2005 during the relief efforts deployed in the region near Aceh, Indonesia (North Sumatra) by Handicap International, one of the 400 NGO that provided humanitarian aid following the tsunami disaster that struck Southeast Asia on December 26, 2004. Working in Banda Aceh and Meulaboh, the author was confronted with the extent of the devastation both in terms of property damage and human suffering. Clinical cases were often severe and rarely encountered in industrialized countries. The tsunami worsened the already poor sanitary conditions: rundown care facilities, poorly trained health care personnel, tropical disease, poor hygiene, and AVP. PMID- 16038347 TI - [Meat inspection. 2.--Bovine]. AB - The purpose of this second technical notice is to provide public health agents with guidelines for inspection of bovine meat. Focusing on hazards related to zoonotic agents, this notice successively presents the symptoms and lesions that must be checked for before approving or rejecting meat for human consumption. PMID- 16038348 TI - [Epidemic of Marburg hemorrhagic fever in Angola]. PMID- 16038349 TI - [Experience of a dermatologist on an overseas military mission: Operation Licorne in Cote d'Ivoire]. AB - The authors report their experience as tropical physicians and as dermatologists within the Medico-surgical group during Operation Licorne an overseas mission with the French Army near Yamussoukro in Cote d'Ivoire. Most of the major endemic diseases were encountered. Only acute infections were treated. The Buruli ulcer epidemic due to Mycobacterioum ulcerans ulcer epidemic is a scourge that mainly affects children with a high incidence of osteitis. PMID- 16038350 TI - [Should National Polio Vaccine Day be eradicated?]. PMID- 16038351 TI - [Osteomyelitis in a Congolese child]. PMID- 16038352 TI - [Unpublished letter from Albert Calmette to Marcel Leger. A new mission for China?]. PMID- 16038353 TI - [Epidemiologic, clinical, and etiologic features of acute delirious episodes (ADE) in Togo: findings of a hospital survey]. AB - This transverse descriptive epidemiologic study carried out in a hospital setting in Togo over a period of 80 days included a total of 63 patients presenting acute delirious episodes. There were 25 women and 38 men ranging in age from 18 to 45 were included. Most patients were young (20 to 34 years) and presented symptoms comparable to those described elsewhere in France and Africa. However several particularites were noted in this fairly protected traditional rural setting in which delirious episodes are uncommon as compared to urban settings in which such cases are more common due to sociocultural stress, drug addiction, and HIV infection. PMID- 16038354 TI - [People living with HIV: daily life and management of the risk of transmission in the infected person's entourage: study in Ouagadougou]. AB - HIV infection has modified social relationships not only within various population subgroups but also at the family level. Patient management has been seriously affected. In 2001 we undertook a descriptive study among a population of PLHIV treated in an ambulatory center in order to 1(o)) describe the socio demographical characteristics of PLHIV admitted to the center and 2(o)) identify the strategies adopted by PLHIV and their families to control the risk of HIV transmission in the entourage. Patients were systematically included over a period of two weeks. Each patient was asked to indicate the name of a family member that could be interviewed by the research team. A total of 188 PLHIV (122 women) ranging in age from 19 to 55 years were questioned. Interviews were conducted in 66 families. Most PLHIV had received some formal education ranging from primary school to university. Over one third (36.4%) had known that they were seropositive for at least one year. Almost three-fourths (73.4%) were aware that HIV was transmissible to other people. The most frequently mentioned transmission hazards were handling contaminated articles (51.5%), unprotected sexual intercourse (46.4%), contact with blood (34.8%), and sharing personal toilet articles (11.6%). The most commonly mentioned preventive measures were abstinence from sex (27.1%), use condoms (28.7%), and protection from pointed or cutting objects (19.1%). According to responding family members, the most common protective measures against HIV transmission were observing hygiene measures (36.5%), discarding any fluids from the ill person in the toilet (26.5%), wearing gloves (25%), and protecting against handling contaminated objects (5.8%). These data suggest that programs aimed at educating PLHIV and their families on the risk of HIV transmission and preventive measure has had a positive impact on the quality of life. PMID- 16038355 TI - [Malaria in an outbreak zone in Oyapock (French Guiana): incidence of malaria attacks in the American Indian population of Camopi]. AB - The Oyapock region is the second largest malaria outbreak area in French Guiana after Maroni. This area that has been less extensively studied that Maroni is characterized by the presence of an exclusively American Indian population on the French bank and by a high incidence of Plasmodium vivax associated with Plasmodium falciparum. The purpose of this study was to determine the incidence of malaria attacks in Camopi, a population center in which most inhabitants live in 15 small villages located along the middle part of the Oyapock River on the Brazilian border. Study involved diagnosis of malaria attacks (ongoing fever or reported fever within 48 hours and presence of asexual Plasmodium parasites with no other apparent etiology) and accurate estimation of its incidence as well as evaluation of the American Indian population. Study was carried out over the two year period between April 2000 and March 2002. The mean annual incidence of malaria was 486 per 1000. Incidence was comparable for the two Plasmodium species: 248 p. 1000 for P. falciparum versus 259 p. 1000 for P. vivax but was much higher in children than adults (402 p. 1000 versus 92 p. 1000). The incidence of P. falciparum varied during the year and from one year to the next. Mixed infection documented by microscopic evidence was uncommon. With an annual incidence of around 500 p 1.000, the Oyapock area of French Guiana must be considered as a malaria hot spot. PMID- 16038356 TI - [Human African trypanosomiasis in the mangrove forest in Guinea: epidemiological and clinical features in two adjacent outbreak areas]. AB - The purpose of this study carried out in two adjacent areas of the coastal mangrove forest of Guinea (Dubreka and Boffa) was to screen the population for disease, provide information on human African trypanosomiasis (HAT, a.k.a. sleeping sickness) and compare the epidemiologic and clinical features with those of outbreak areas in the Ivory Coast where more data is currently available. Cases of HAT were confirmed by parasitological testing after active medical work up (91 of 9637 patients examined). Five cases were confirmed in patients in treatment centers. Of the first 57 cases admitted for treatment in the Dubreka and Boffa centers, 29 were responded to a clinical and epidemiological questionnaire and underwent thorough clinical examination. Disease stage was determined by cytochemical testing of cerebrospinal fluid. As in outbreak areas of the Ivory Coast, sleeping sickness in Dubreka and Boffa is a rural disease mainly affecting the working population. Most cases identified in Guinea involved men and women working in farming, fishing, or salt extraction. However unlike Ivory Coast outbreak areas where ethnic diversity related to share cropping is considered to play a major role in maintaining endemicity, almost all patients in our study (98%) were from the native Soussou population that is self employed and lives in villages with no immigrant population. While clinical symptoms observed in these patients were not different from those reported elsewhere, there was a high frequency of cervical adenopathy (93%). This finding could provide a useful diagnostic sign for screening populations living in these mangrove forest regions and as a source for parasitological diagnosis as shown by the fact that 88.5% of patients were screened on the basis of lymph node fluid specimens. Most patients including among those identified by active work-up (5%) were in the meningo encephalitis phase of the disease (98%). The findings of this study underline the need not only to continue surveillance in these regions but also to extend surveillance throughout the country as a means of avoiding recrudescence and extension of the disease. PMID- 16038357 TI - [Pulmonary bilharziosis due to Schistosoma haematobium: pitfalls of species diagnosis. A case report from Libreville, Gabon]. AB - The purpose of this report is to describe a case of pulmonary schistosomiasis treated at la Fondation Jeanne Ebori in Libreville, Gabon. This case is exceptional due to the rarity of the disease and the schistosomiasis agent involved. The patient was a 47-year-old woman who presented with recurrent right sided pneumonia 6 months after initial hospitalization and nonspecific antimicrobial therapy. Her general status was altered by hyperthermia, right chest pain, and repetitive bouts of hemoptysia. Because etiological diagnosis could not be achieved and clinical condition was deteriorating, the decision was taken to perform exploratory thoracotomy. Based on operative findings, medial and lower lobectomy was performed. Histological examination demonstrated granulomatous inflammatory lesions due to bilharziosis. Infection was attributed to Schistosoma haematobium that is the most common agent in pulmonary localizations. However Ziehl coloration raised the possibility of involvement of Schistosoma intercalatum that has never been observed in the lung and of a hybrid species. Natural hybridization between S. haematobium and S. intercalatum and the presence of active transmission of the Schistosoma hybrid has been suspected in Gabon. PMID- 16038358 TI - [Utility and limitations of laboratory diagnosis of amebiasis]. AB - Entamoeba histolytica is an invasive and pathogenic protozoan parasite that causes amebiasis. It must be distinguished from Entamoeba dispar, a nonpathogenic commensal parasite of the human gut lumen that is morphologically identical to Entamoeba histolytica. Diagnosis of amoebic colitis currently requires combination of microscopic examination of stool specimens with another technique allowing positive identification of the two species. Stool culture followed by zymodem analysis is considered as gold standard but is not applicable in routine practice. Detection of specific Entamoeba histolytica antigens in stools is a fast, sensitive technique that should be considered as the method of choice. Stool PCR is a highly sensitive and specific technique but high cost make it unsuitable for use in endemic areas where economic conditions are difficult. The utility of serologic tests in distinguishing Entamoeba dispar from Entamoeba histolytica is controversial. However serology is still considered as the method of choice for diagnosis of extraintestinal amebiasis. Circulating Gal/GalNac lectin antigens can be detected in the serum of 96% of patients with untreated amoebic liver abscess. In the future this method should allow early diagnosis and treatment of extraintestinal amoebiasis in patients who have not yet developed detectable serum antibodies. PMID- 16038359 TI - Contribution of PCR-based methods to diagnosis and management of imported malaria. AB - Since the first description, in 1990, of the diagnosis of Plasmodium falciparum infection by polymerase-chain-reaction (PCR), the role of this kind of molecular method in laboratory diagnosis of imported malaria is still a topical question. Various molecular assays have been used, the first of which was hybridization using labeled probes in 1984. When compared to thick blood smear, this test displayed a sensitivity ranging from 65% to 81% and specificity was close to 100%. The next technical improvement was the introduction of the so-called polymerase chain reaction (PCR), the principle of which was described in 1985. In 1993, a PCR-based assay detecting all four Plasmodium species was published, followed by different variants of this method. By the turn of the century, novel real-time PCR slashed workaround time, which dropped from 2 1/2 hours to less than 1 hour. Moreover, automatic reading with no human action on PCR products reduced the risks of contamination. The first application of real-time PCR to the diagnosis of malaria was published in 2001. PCR-based assays were found to be more sensitive than all conventional methods. Variations in sensitivity were probably due to different medical practices as well as to the proportion of various types of subjects (travelers under chemoprophylaxis, immigrants from malaria-endemic areas) in the population undergoing malaria diagnosis. The target of the primers was also of crucial importance: for the detection of P. falciparum, the most efficient assays amplified either the gene SSUrRNA, or Pf155/RESA, or Cox 1. Specificity of PCR results is guaranteed by the nature of the target for primers or probes, as determined by the studies of the Plasmodium genome whose results are available in GenBank. PCR use often corrected the results of Plasmodium species identification by microscopy and PCR-based methods were found to be the most efficient for the detection of mixed infections. Concerning the diagnosis of imported malaria, it appears clearly that PCR should be considered as second-line method which can be especially interesting, as a negative result rules out malaria in febrile patients. However, the use of PCR assays appears to be restricted to health centers, such as University Hospitals, for whom malaria identification is an important and routine problem. In the future, the detection of mutations related to drug resistance could be used to orient anti-malarial therapy. PMID- 16038360 TI - [Monitoring and evaluation of health care programs in Subsaharian Africa]. AB - The monitoring and evaluation is usually the weakest component of health programs in sub-Saharan Africa, what is undermining the sustainability of funding. The problems are complex and the weaknesses of the health systems are reflected on the monitoring and evaluation of specific programs. This paper gives an insight of the problems faced during field missions for monitoring and evaluation. The steps for building the M&E system have been reviewed and keys points for implementation have been provided. PMID- 16038361 TI - [Assessment of transfusion requirements: a way to in improve perioperative management of blood products?]. AB - Autogenic blood transfusion is indispensable in emergency surgery situations involving severe blood loss. It may also be required in some non-emergency surgical and obstetrical situations. The use of blood-sparing techniques as an alternative to autogenic blood transfusions blood loss can be especially beneficial in tropical settings where the risk of viral transmission is high. The combined use of blood-sparing and autogenic transfusion techniques requires preoperative assessment of transfusion requirements. The expected amount of preoperative blood loss must be determined and compared with the acceptable amount of blood loss for the patient in function of transfusion threshold. Various techniques to reduce the need for autogenic blood transfusion can be used depending on locally available resources. Blood-sparing techniques include treatment to increase the patient's baseline hemoglobin rates, use of cell saving systems for autologous blood transfusion, and/or perioperative transfusion of recuperated blood. In this article these techniques are illustrated in two practical clinical cases. PMID- 16038362 TI - [The unfinished agenda for child survival: what role for the integrated management of childhood illness?]. AB - The endorsement by the United Nations General Assembly of the Millennium Development Goals (MDG) and the growing acknowledgment by the international community that child survival is an unfinished agenda created a new momentum for rapid scaling up of effective child health interventions. In this review, the authors discuss the environment in which child health programs are being implemented and the potential role of the integrated management of childhood illness (IMCI) strategy in country efforts to achieve the MDGs. The discussion is based on the conclusions of a multi-country analytic review of the IMCI strategy conducted jointly by DFID, UNICEF, USAID, and WHO as well as the results of another multi-country evaluation coordinated by the WHO on IMCI costs, effectiveness, and impact. The article concludes on the need to increase child health investments and on the potential importance of IMCI in improving child survival. However, the MDGs may not be reached if IMCI is not implemented in conjunction with other strategies to reduce mortality during the first days of life and to strengthen the health system. The authors also stress the need to increase research on mechanisms to scale up delivery of existing public health interventions. PMID- 16038363 TI - [The sarcoidosis in Cambodia: a poorly understood disease]. PMID- 16038364 TI - [Role of formal education in the fight against female excision in Senegal]. PMID- 16038365 TI - [Community participation in NVD against poliomyelitis in Kanel, Senegal]. PMID- 16038366 TI - [Seventh expert's meeting on envenomation by poisonous animals]. PMID- 16038367 TI - [Enterovirus meeting (St Etienne, October 12-13, 2004)]. PMID- 16038368 TI - Colorectal mesenchymal tumor: a clinicopathologic study of 25 cases. AB - BACKGROUND: It is important to distinguish gastrointestinal stromal tumors (GISTs) from other gastrointestinal mesenchymal tumors (GIMTs), because of the malignant potential of GISTs and the availability of molecular targeted therapy. GISTs represent the most common subgroup of GIMTs, and rarely occur in the colon and rectum. The first objective of our retrospective study was to reclassify colorectal mesenchymal tumors, from files collected over 20 years, to determine if, based on immunohistologic features, the lesions were truly GISTs. The second objective was to identify the relationship between clinicopathologic features and prognostic factors of GISTs in the colon and rectum. METHODS: We evaluated all cases of colorectal mesenchymal tumor identified from the database of the Department of Surgical Pathology at Taichung Veterans General Hospital for the period 1983-2001. For 25 patients, clinical data, and information about tumor characteristics, surgical procedures, and survival outcomes, were obtained and analyzed. Histopathologic evaluations, and appropriate immunohistochemical markers, were used to distinguish between various GIMT subtypes. The relationship between KIT expression and clinicopathologic features was investigated. RESULTS: The following variables were significantly associated with different CD117 results: symptomatic presentation, location, gross features, tumor size, mitotic count, cellularity, and type of surgery. Only 18 tumors were identified as GISTs. For these, the following variables were significantly associated (by univariate analysis) with increased lethality: tumor size (p = 0.049); mitotic count (p = 0.019); nuclear atypia (p = 0.019); and tumor necrosis (p = 0.045). However, only mitotic activity showed a significant difference in the survival analysis (p = 0.0304; log-rank test). CONCLUSION: Two clinicopathologically different categories were identified from our colorectal mesenchymal tumors: intramural GISTs and polypoid submucosal leiomyomas. Our study suggests that GIST is a better categorization than smooth muscle tumor because of the malignant potential. Prognosis is strictly related to the number of mitoses. However, tumor size, nuclear atypia and tumor necrosis are probably also significant predictive factors of lethality. Future studies with DNA analysis and larger patient numbers are essential to evaluate the prognostic significance of our findings. PMID- 16038369 TI - Clinical factors and outcomes in patients with acute mesenteric ischemia in the emergency department. AB - BACKGROUND: The purpose of this study was to determine the initial clinical characteristics of acute mesenteric ischemia and identify variables associated with adverse outcomes in the emergency department (ED). METHODS: The charts of 124 consecutive patients with surgically and pathologically identified acute mesenteric ischemia from September 1990 to September 2000 were reviewed retrospectively to obtain data about demographics, initial clinical presentations, predisposing diseases, previous medications, laboratory tests, and common findings on computed tomography scans with contrast. Only patients admitted through the ED and treated on medical or surgical wards were enrolled. RESULTS: Mean patient age was 71.1 years (range, 25-100 years). The overall mortality rate was 50%. There were no significant differences in gender, underlying disease, previous medication, initial signs and symptoms, and causes of mesenteric infarction, between survivors and non-survivors. Univariate analysis demonstrated that older age, bandemia, hepatic and renal impairment, hyperamylasemia, metabolic acidosis, hypoxia, intramural pneumatosis, and septic syndrome, were more frequent in patients who died than in those who survived (p < 0.05). Logistic regression identified the following variables as independent predictors of death: old age (odds ratio, OR, 1.077; 95% confidence interval, Cl, 1.013, 1.146; p = 0.02); bandemia (OR, 3.894; 95% Cl, 1.160, 13.074; p = 0.03); elevated serum aspartate aminotransferase (AST; OR, 4.532; 95% Cl, 1.274, 16.122; p = 0.02); increased blood urea nitrogen (BUN; OR, 7.219; 95% Cl, 1.166, 44.696; p = 0.03); and metabolic acidosis (OR, 6.604; 95% Cl, 1.804, 24.171; p < 0.01). CONCLUSION: A high index of suspicion and aggressive diagnostic imaging can facilitate early diagnosis and improve outcomes for patients with acute mesenteric ischemia. Risk stratification showed that elderly patients with metabolic acidosis, bandemia, or elevated AST and BUN had a poor prognosis. Greater therapeutic intervention is advocated to reduce mortality in high-risk patients with acute mesenteric ischemia. PMID- 16038370 TI - A randomized, parallel, comparative study of the efficacy and safety of nafarelin versus danazol in the treatment of endometriosis in Taiwan. AB - BACKGROUND: The purpose of this study was to evaluate the efficacy and safety of nafarelin, a gonadotropin-releasing hormone (GnRH) analogue, versus danazol in the treatment of women with endometriosis in Taiwan. METHODS: Fifty-nine women with laparoscopically and pathologically confirmed endometriosis were randomized to receive nafarelin or danazol for 180 days. Efficacy was assessed from mean changes in laparoscopy score (LS) and total symptom severity score (TSSS). Adverse events (AEs) and laboratory parameters, including hematology, hepatic function, blood pressure, and lipid levels, were monitored for safety evaluations. RESULTS: All demographic and baseline factors, except body weight, were comparable between the 2 treatment groups. Both nafarelin and danazol satisfactorily resolved pelvic tenderness, induration, pelvic pain, dysmenorrhea and dyspareunia. No significant differences were noted in efficacy endpoints between nafarelin and danazol regarding LS and TSSS at 90 and 180 days of treatment. No significant difference was observed between the 2 groups regarding the overall incidence of AEs, except for laboratory-related AEs. However, nafarelin tended to have less impact than danazol on aspartate transaminase and alanine transaminase, and nafarelin was better tolerated than danazol regarding changes in lipid profiles. Both treatments had little or no effect on hematologic parameters. CONCLUSION: Nafarelin and danazol demonstrated similar clinical efficacy, but nafarelin was associated with fewer laboratory changes and a stable lipid profile, relative to danazol. Moreover, intranasally administered nafarelin is noninvasive, and may be a more comfortable and safer alternative to slow release injectable GnRH agonists. Based on this study, we suggest that nafarelin, like other GnRH analogues, may be a treatment of choice for Taiwanese women with endometriosis. However, direct comparative studies of nafarelin with slow-release injectable GnRH agonists are now required. PMID- 16038371 TI - Radiosurgery for vestibular schwannomas. AB - BACKGROUND: Radiosurgery has been established as an important alternative to microsurgery. We report our experience with radiosurgery for tumor control and the complications of unilateral vestibular schwannomas. METHODS: We reviewed our early experience regarding clinical presentation, management and outcomes in 45 patients with acoustic schwannomas who underwent gamma knife stereotactic radiosurgery. The median follow-up period was 25 months (range, 6-48 months). Thirteen patients had undergone 1 or more previous resections before radiosurgery; 32 underwent radiosurgery as the first procedure. Median tumor volume was 4.5 mL (range, 0.5-30.0), and median radiotherapy dose was 11.5 Gy (range, 10.5-14.0 Gy). RESULTS: Tumor control was achieved in 43 patients (95.6%). Loss of central contrast enhancement was a characteristic change and was noted in 29 patients (64.4%). Reduction in tumor size was shown in 15 patients (33.3%). Thirteen patients (28.9%) had good or serviceable hearing preoperatively, and in all of these, the preoperative status was retained immediately after radiosurgery. At follow-up, however, 10 patients (76.9%) had preserved hearing and 3 (23.1%) had reduced hearing on the treated side. Hearing in 1 patient that was not serviceable preoperatively later improved to a serviceable level. No patients had delayed facial palsy or lower cranial nerve dysfunction, but one had delayed trigeminal sensory loss. CONCLUSION: Radiosurgery achieved a high tumor control rate and a relatively low post radiosurgical complication rate for acoustic neuromas. PMID- 16038372 TI - Risk factors for retinal detachment after cataract surgery in southern Taiwan. AB - BACKGROUND: This study was designed to investigate potential risk factors for retinal detachment (RD) after cataract surgery in southern Taiwan. METHODS: This was a prospective cohort study. Submitted charts and claimed records for insurance beneficiaries who underwent cataract extraction between August 1999 and December 2001 were collected from the Bureau of National Health Insurance. Data from these records were examined, including demographic characteristics, past history, refractive status, axial length, and type and complications of cataract extraction. At the end of 2002, any ophthalmologic diagnosis and related treatments, based on procedure and diagnosis codes listed in physician bills, were evaluated. RESULTS: A total of 9,398 patients were evaluated. The cumulative risk of RD in our study group was 0.4% at the end of follow-up. The mean follow up time was 25.5 +/- 7.7 months. Age had a significant influence on the risk of RD after cataract surgery (p = 0.006), whereas gender did not. Axial length also had a significant effect on the risk of RD after cataract extraction (p < 0.001), whereas systemic disease or the type of operation did not correlate significantly with RD. Intraoperative complications had a significant influence on RD risk only in patients aged more than 60 years (p = 0.042). Neodymium:yttrium-aluminum garnet (Nd:YAG) laser posterior capsulotomy significantly reduced the risk of RD only in patients aged 50 years or younger (p < 0.001). CONCLUSION: Our results show that young age and long axial length are both significantly associated with RD after cataract surgery. A more restrictive attitude towards early cataract extraction may therefore be appropriate, especially in highly myopic patients. PMID- 16038373 TI - Intestinal obstruction in patients with previous laparotomy for non-malignancy. AB - BACKGROUND: Intestinal obstruction is one of the most common surgical emergencies. The aim of this study was to identify important management information from the evaluation of patients with intestinal obstruction who had undergone previous laparotomy for non-malignancy. METHODS: Data from 176 patients with previous laparotomy for non-malignancy, and who were operated on for intestinal obstruction, were collected and analyzed retrospectively. RESULTS: Gastroduodenal operations, appendectomy, and obstetric/gynecologic procedures were the 3 most common previous abdominal surgeries. More than half of all bowel obstructions developed within 10 years after previous laparotomy, and particularly within the first 5 years. Most obstructions were related to adhesion, although their etiologies were diverse. The rate of bowel strangulation was much higher in patients with internal herniation, volvulus, intussusception, closed loop, and diaphragmatic hernia than in patients with simple adhesion, bezoar, tumor, and inflammation (48.3% vs 12.2%). The surgical mortality rate correlated significantly with bowel strangulation: the overall rate was 6.8%, that in patients with strangulation was 18.8%, and that in patients without strangulation was 4.2%. CONCLUSION: The etiologies of intestinal obstruction were not only significantly related to bowel strangulation, but were also an important determinant of therapeutic strategy. PMID- 16038374 TI - Temporal arteritis. AB - Temporal arteritis, a chronic inflammatory vasculitis involving medium- and large sized arteries, has rarely been reported in Asia. However, we report 2 cases, in which the patients initially presented with headache. Physical examination disclosed engorged, hard and palpable vessels in the temporal areas. Temporal artery biopsy revealed 2 different types of arteritis: the multinucleated giant cell type and the panarteritis type without multinucleated giant cells. One patient was positive for immunoglobulin G anticardiolipin antibody. The pathologic findings of the different subsets of temporal arteritis, and the relationship between anticardiolipin antibody and the extent of vascular complications of temporal arteritis, are discussed. PMID- 16038375 TI - Pregnancy complicated with pulmonary edema due to hyperthyroidism. AB - Hyperthyroidism is one of the most common causes of cardiac failure. Blood volume expands greatly during pregnancy, especially after the last part of the second trimester. Such expansion exacerbates the symptoms of heart failure and accelerates the development of pulmonary edema when abnormal thyroid function is not well controlled. Two cases of pregnancy complicated with congestive heart failure and pulmonary edema due to hyperthyroidism are reported here. Both patients did not receive treatment for hyperthyroidism during pregnancy, and both sought clinical help during the third trimester. The clinical problems were resolved by medical management before delivery. PMID- 16038376 TI - Sarcoidal granuloma in cervical lymph nodes. AB - Sarcoidosis is a multiorgan granulomatous disease, the most common head and neck manifestation of which is cervical lymphadenopathy. Only the presentation of sarcoidal granuloma in cervical lymph nodes without typical manifestations of systemic sarcoidosis poses a diagnostic difficulty. We describe the case of a 39 year-old male who had a 2-month history of a progressively increasing mass with soreness in his right neck. The biopsy from the neck mass demonstrated non caseating epithelioid cell granuloma of the lymph nodes. The differential diagnoses of mycobacterial or fungal infections were excluded. Thoracic evaluations, including chest X-ray and high-resolution computed tomography, revealed no abnormal findings. Treatment with systemic corticosteroids resulted in improved clinical symptoms. No recurrence of the neck mass or other signs of systemic sarcoidosis were noted during 1.5 years of follow-up. Although our patient's definitive diagnosis could not be determined, the case highlights 2 important issues: sarcoidal granuloma in lymph nodes may be a precursor of sarcoidosis, even in the absence of pulmonary or other systemic involvement; and regular follow-up is recommended in such cases. PMID- 16038377 TI - Shepherd's crook deformity of polyostotic fibrous dysplasia treated with corrective osteotomy and dynamic hip screw. AB - Fibrous dysplasia, a condition in which the skeleton fails to develop normally, is characterized by fibroblastic stroma and immature bone. Bowing of the long bones occurs frequently in the polyostotic form, and stress fractures often result. Shepherd's crook deformity is a characteristic feature of fibrous dysplasia. The goal of its treatment is to obtain normal walking ability and relieve pain due to pathologic fracture secondary to the deformity; however, correction of the deformity is a surgical challenge. We present 2 cases of shepherd's crook deformity treated with corrective osteotomy and a dynamic hip screw. Both cases showed good bone healing and no recurrent deformity. The gross deformities were corrected, and both patients were pain-free after operation. PMID- 16038378 TI - Immunological and ultrastructural characterization of plasma cells of human periapical chronic inflammatory lesions (granulomas). AB - Despite many studies on the topic, plasma cells found in human periapical chronic inflammatory lesions (granulomas) continue to present unresolved issues. In this study, we tried to assess quantitatively and qualitatively the nature of plasma cells of 4 human periapical granulomas. Samples were analyzed for relative amounts of IgG-, IgM-, IgA-, and IgE-positive plasma cells by immunohistochemistry, and for morphological changes by transmission electron microscopy (TEM). By immunohistochemistry, many plasma cells stained positively with anti-IgG and anti-IgM antibodies; fewer cells reacted with anti-IgE and anti IgA. Russell Bodies, controversial aspects of plasma cell maturation, showed positive reactivity of the superficial layer only to antibodies against IgG and IgM. By TEM analysis, phenotypes of normal and dysfunctional plasma cells (Mott cells) were evident. Russell Bodies appeared as intra- or extracellular round vesicles, with an homogeneous internal core, and an external membrane, resembling rough endoplasmatic reticulum (RER). We can conclude that mucosal immune response is not the predominant type in the periapical lesions examined. Positive immunoreaction for IgG and IgM of Russell Bodies may be due to the residual RER membrane, whereas components of yet unidentified nature may occupy the internal core. PMID- 16038379 TI - Interstitial cells of Cajal: once negligible players, now blazing protagonists. AB - Cajal in 1889 described a network of anastomosing interstitial cells in the gut muscle coat and hypothesized that they were accessory primitive neurons exerting a direct regulatory effect on smooth muscle contraction. Reticularists (among them Golgi) sustained that this net was not an assembly of individual cells but a true syncytium and the foremost dissidents, such as Kolliker and Dogiel, declared they were connective tissue cells. Keith, the discoverer of the sino-atrial node, suggested that these cells "constitute a pacemaker system of the intestinal muscle". In the period 1925-1960, there were papers still discussing the role and nature of the interstitial cells. The majority of these papers, however, reflect the fight between neuronists and reticularists. Around 1960, the reality of the neurons was established by ultrastructural evidence and interstitial cells degraded to fibroblasts or Schwann cells. By 1970, electron microscopists began to pay attention to these cells (from now named ICC). Among them, I myself concluded that ICC have smooth muscle features and might well be pacemaker cells. In this period, vital methylene-blue staining followed by electron microscopy firmly identified the ICC as myoid cells and the zinc iodide-osmic acid method, used to stain neurons, was also excellent for ICC and, when applied for electron microscopy, confirmed the identity of these cells. In the meantime different ICC populations were found in the gut muscle coat with region-specific location and region-specific features. By 1980, ICC, revealing themselves as myoid cells, a nature far more exciting than former ones, underwent to a booming interest and also physiologists began to study them. At present, it has been proved that one population, distributed throughout the entire gut, plays a pacemaker role; a second population, located intramuscularly in the stomach, is involved in neurotransmission, and a third population, specific of the small intestine, is part of the intestinal stretch receptor. By 1980 up to day, the differentiating steps of these cells were studied and factors implied in their maturation during foetal life and in the maintenance of their differentiated state in adulthood were identified. There has been also a rapidly evolving knowledge of specific molecules which are expressed on ICC, some of which useful for ICC identification under light- and electron microscope with a relative facility, some functionally implicated in neurotransmission and others in metabolic pathways strictly related to specific ICC behaviours. The more recent studies are considering the possibility of an ICC plasticity, transdifferentiation and apoptosis, especially in view of a direct implication of these cells in certain disorders of gut motility. Perspectives for future research are mainly concerning ICC alterations in gastrointestinal diseases. PMID- 16038380 TI - Signs of normal proliferation in the telencephalon of adult male songbirds (serinus serinus), as shown by PCNA-positivity. AB - The immunocytochemical expression of Proliferating Cell Nuclear Antigen (PCNA) (a cycline that coadjuvates DNA polymerase delta) becomes appreciable in the cell cycle when DNA synthesis occurs; hence cells in the S phase can be revealed by means of monoclonal antibodies. Therefore, PCNA can be considered a marker of proliferation, and numerous literature reports have demonstrated the reliability of the PCNA test. Since normal neurogenic events can still occur in the brain tissue of adult homeothermic vertebrates (especially songbirds), we evaluated if the persistence of spontaneous proliferation could be revealed in adult male songbirds (Serinus serinus) using the PCNA marker, the same test we used previously to study the persistence of natural proliferation in the encephalon of adult heterothermic vertebrates. The patterns of PCNA positivity showed normal proliferation in the telencephalon of the adult male Serinus serinus. This activity was shown by cells interposed among the epithelial cells lining the lateral side of each ventricular cavity, both in correspondence to the apical tracts and declivities of the ependyma and arranged, here and there, either in groups or slightly separated. As in our previous studies on PCNA expression and persistence of spontaneous encephalic proliferation in adult poikilothermal vertebrates (in the telencephalon of Podarcis, Triturus and Rana, and in the telencephalon, mesencephalon and cerebellum of Carassius), the results of the present research largely agree with the findings of previous Authors, usually obtained with different methods. This agreement confirms the reliability of the PCNA test used for this type of investigation. PMID- 16038381 TI - The relation between the acupoint structures and the clinical therapeutic effects. AB - We performed a histological study upon the acupuncture points and their effectiveness of clinical treatment; the background was the clinic evidence that a multitude of points are used in treating a disease, but only some of which may have an efficacy, since the others did not. After a comparative histological and anatomic study, it comes out that those points, which are more effective from a structural point of view, identify a neural fibrillar concentration, a well developed capillary network and an increased mucopolysaccharides (MPS) concentration, in particular, acid mucopolysaccharides. The present paper presents histological data, which demonstrate the difference in the structure of the acupuncture points, postulating their specific influence on clinical treatment. PMID- 16038382 TI - Acupoints [corrected] and meridians: a histochemical study. AB - The aim of this study was to elucidate the relationship between the structural specificities of acupoints and meridians as well as their clinical effects. We processed 356 specimens, 287 of which from 48 adult and 2 newborn cadavers and the remaining 69 from living patients; samples were taken at three different levels: (1) beneath acupoints; (2) between meridians; (3) at a distance from meridians. We performed seven different staining to show the distribution of collagen fibers, reticular fibers, mucopolysaccharides (MPS), connective tissue, nerve threads, and blood vessels in specimens obtained from different areas. We found that some structural and biochemical discrepancies associated with acupoints and meridians including: (1) mucopolysaccharides (MPS), in particular acid MPS; (2) collagen fibers; (3) nerve endings. We discussed these findings from an anatomo-clinical point of view. PMID- 16038383 TI - Performance of polymethyl pentene oxygenators for neonatal extracorporeal membrane oxygenation: a comparison with silicone membrane oxygenators. AB - OBJECTIVE: To review the performance of polymethyl pentene versus silicone oxygenators in terms of efficiency in priming and oxygenation, oxygenator resistance, requirements for coagulation proteins and consumption of blood products, for neonatal extracorporeal membrane oxygenation (ECMO) patients. STUDY DESIGN: Forty consecutive neonates were selected retrospectively pre- and post introduction of the new polymethyl pentene (PMP) oxygenators. They formed two equal groups. After calculation of the sample size, data were collected from ELSO registry forms and patient records. Results were analysed using parametric and non-parametric tests. RESULTS: Neonatal PMP (N-PMP) oxygenators were smaller, faster and easier to prime. They were less efficient than silicone oxygenators, especially in carbon dioxide elimination, and, therefore, required higher sweeps. The preservation of coagulation proteins was significantly better, but there was no reduction in the consumption of blood products, despite having less than half the surface area and significantly lower blood path resistance. CONCLUSION: Small PMP oxygenators (Medos Hilite 800 LT) provide adequate gas exchange and offer technical advantages in terms of more efficient priming, reduced haemodynamic resistance and better control and preservation of coagulation proteins than silicone oxygenators. PMID- 16038384 TI - Comparison of hollow-fiber membrane oxygenators in terms of pressure drop of the membranes during normothermic and hypothermic cardiopulmonary bypass in neonates. AB - The objective of this study was to investigate the effects of two hollow-fiber membrane oxygenators, the Capiox SX10 and the Lilliput 901, on pressure drop of the membranes during normothermic and hypothermic cardiopulmonary bypass (CPB) in neonates. METHODS: Twenty-six congenital heart surgery patients (n = 13 in each group) with a mean weight of 3 kg were included in this study. Pressure drops of the membranes, pre- and post-oxygenator extracorporeal circuit pressures (ECC) were recorded during normothermic CPB, hypothermic CPB (20 degrees C) and after rewarming. There were no differences between the groups in mean arterial pressure, pump flow rate, temperature, duration of CPB, crossclamp time or the severity of the surgical repairs. RESULTS: Pressure drop of the Capiox SX10 oxygenator was significantly lower during normothermic (32 +/- 10 versus 55 +/- 16 mmHg, p < 0.001), hypothermic (38 +/- 15 versus 72 +/- 18 mmHg, p < 0.001) and post-rewarming (42 +/- 13 versus 72 +/- 21 mmHg, p < 0.001) periods compared to the Lilliput oxygenator. In the Capiox group, the pre-oxygenator ECC pressure was also significantly lower during normothermic CPB (142 +/- 27 versus 184 +/- 43 mmHg, p < 0.01), hypothermic CPB (162 +/- 30 versus 199 +/- 38 mmHg, p < 0.01) and after rewarming periods (172 +/- 32 versus 212 +/- 42 mmHg, p < 0.01). Post oxygenator pressures in the Capiox group were also lower than in the Lilliput group, but results were not statistically significant. CONCLUSIONS: These results suggest that the Capiox SX10 hollow-fiber membrane oxygenator produced significantly lower membrane pressure drops and pre- and post-oxygenator ECC during normothermic and hypothermic CPB. Thus, blood trauma with the Capiox during extracorporeal circulation may be significantly lower compared to the Lilliput. Further studies, including the level of complements, platelets, neutrophils and cytokines, with these oxygenators are warranted. PMID- 16038385 TI - A survey for pain and sedation medications in pediatric patients during extracorporeal membrane oxygenation. AB - Routine administration of large amounts of pain and sedative medication is common to critically ill pediatric patients undergoing extracorporeal membrane oxygenation (ECMO) for cardiopulmonary failure. It has been our experience that pediatric patients are the most difficult age group in which to achieve an ideal pain and sedative control due to the narrow margin of safety. The purpose of this study was to determine the general practice guideline used for pain and anxiolytic pharmacotherapy for pediatric patients at ECMO centers. We sent a survey questionnaire to all ECMO centers in the USA that treat pediatric respiratory failure patients. Of the 46 responding centers (including telephone follow-ups), 37 (80%) centers had an active pediatric ECMO programs for patients with severe respiratory failure. Fentanyl was the most commonly used pain medication and continuous infusion, administered directly to the patient, was preferred. Subjective effectiveness of various pharmacological agents was variable without clear consensus; however, midazolam was considered to be the most effective agent used. PMID- 16038386 TI - Microbubble production in an in vitro cardiopulmonary bypass circuit ventilated with xenon. AB - Xenon, as an anaesthetic gas, has the potential to be used in an increasing range of applications. However, its use in cardiopulmonary bypass (CPB) has not yet progressed from the rat model due to concerns that its relative insolubility may cause microbubble formation and/or expansion in the micro-vasculature of the patient. An in vitro CPB circuit was designed to create and measure gaseous microbubbles over a range of temperature gradients, pressure drop and gas tensions. We were able to demonstrate that our test circuit did not produce any significant microbubbles and that, under normal physiological blood pressures, a fixed gas bubble in connection with the circuit did not grow in the presence of Xe. PMID- 16038387 TI - Use of a dynamic bubble trap in the arterial line reduces microbubbles during cardiopulmonary bypass and microembolic signals in the middle cerebral artery. AB - Neurological complications remain an important cause of morbidity and mortality of patients following cardiopulmonary bypass (CPB). Microemboli, as well as cerebral hypoperfusion, are the main postulated mechanisms. This study demonstrates that the insertion of a dynamic bubble trap (DBT) into the curcuit reduces microbubbles in the arterial line and microembolic signals (MES) in the middle cerebral arteries (MCAs). We investigated 12 patients during coronary artery bypass grafting (CABG). The DBT was inserted between the arterial filter and the arterial cannula. For detection of microemboli before and after the DBT, a special ultrasound Doppler device was used. MES were detected by transcranial Doppler monitoring in both MCAs of the patients. Microbubbles and MES were counted during bypass. These data were compared to 12 patients who were operated in a previous period without the use of a DBT. There were no significant differences in both groups with respect to gender, age, crossclamp and bypass time and number of anastomoses. In the group without a DBT in the circuit, a mean of 6311 microbubbles per operation could be observed distal to the arterial filter, corresponding to 282 MES. After inclusion of a DBT, we could register, in the second group, 8496 microemboli proximal and 2915 distal of the DBT, corresponding to 89 MES per operation. The reduction rate of microbubbles in the tubing was 65.7%, corresponding to a reduction in MES of about 86.2%. We conclude that the insertion of a DBT in the arterial line of CPB circuit protects the cerebrovascular system from microembolic events, as demonstrated by lower MES counts. PMID- 16038388 TI - Hypothermia-induced platelet aggregation and cognitive decline in coronary artery bypass surgery: a pilot study. AB - Hypothermia-induced platelet aggregation (HIPA) was previously reported in whole blood exposed to synthetic surfaces at 24 degrees-32 degrees C in one-third of normal subjects tested. Cardiopulmonary bypass, conducted with hypothermia, may lead to such aggregation, resulting in microvascular occlusion contributing to cognitive impairment. This pilot study was conducted to explore the relationship between HIPA and cognitive outcome at hospital discharge in patients undergoing coronary artery bypass graft (CABG) surgery as a first step toward a longer-term study. Patients (n = 45) undergoing mild to moderate hypothermia (32 degrees-28 degrees C) during CABG surgery underwent cognitive testing preoperatively and prior to hospital discharge. Tests included: visual and verbal memory, mental processing speed, executive function, language, and intellectual function. HIPA was identified using an in vitro assay in which blood flowing in polyvinychloride tubing was subjected to hypothermia, and platelet aggregates were detected using microscopy and passing the exiting blood through a 20-microm pore filter. Forty four percent of patients exhibited HIPA. The entire cohort exhibited significant postoperative cognitive decline in verbal memory, mental processing speed and executive function. There was greater cognitive decline in the group with HIPA compared with the group not exhibiting this phenomenon. The patients with HIPA showed significant decline in four of five cognitive measures whereas patients not exhibiting this phenomenon declined in only two of five cognitive measures. HIPA appears to be associated with an added risk of cognitive decline immediately following CABG surgery employing mild to moderate hypothermia. The findings of our study suggest the long-term cognitive effects of hypothermia-induced platelet aggregation need to be explored. PMID- 16038389 TI - Procalcitonin is useful whereas C-reactive protein is not, to predict complications following coronary artery bypass surgery. AB - BACKGROUND: The respective value of procalcitonin (PCT) and C-reactive protein (CRP) as markers of postoperative complications after coronary bypass surgery is unclear. Therefore, complications during one week after surgery were studied to evaluate the predictive role of PCT and CRP changes during the immediate postoperative period. METHODS: Thirty-two patients, in whom an uneventful immediate postoperative course was anticipated, were prospectively enrolled and followed-up to the 7th postoperative day. At the end of the follow-up, patients were divided into two groups. Group A were patients with an uncomplicated postoperative course and Group B were patients with a complicated postoperative course. RESULTS: Serum samples were drawn for PCT and CRP determination after induction of anesthesia (baseline), at the end of surgery and daily until postoperative day 2. Baseline serum PCT concentrations were 0.11 +/- 0.09 and 0.20 +/- 0.21 ng/mL in Groups A and B, respectively (NS). Serum PCT concentration increased compared with baseline in both groups during the first two days after surgery. The increase in serum PCT concentration was significantly greater in Group B than A patients (p < 0.0002). Considering a perioperative abnormal cut off value of >0.5 ng/mL, there were none in Group A versus 57% in Group B (p < 0.0001). Baseline serum CRP concentrations were 1.44 +/- 1.30 and 1.58 +/- 1.35 ng/mL in Groups A and B, respectively (NS). After surgery, CRP increased significantly compared with baseline in both groups. When changes in time-varying variables were included in a logistic model, complications were predicted by changes (between baseline and end of surgery values) of PCT (coefficient = 9.410; t = 2.18) and heart rate (coefficient = 0.075; t = 1.57), whereas changes of CRP, white blood cells, mean blood and central venous pressures did not contribute statistically. The model constant was -4.827 (t = -2.43) and the ROC curve area was 0.8971. Thus, absolute PCT changes of 0.20, 0.40 and 0.60 ng/mL carry an approximate risk of 5, 26 and 69%, respectively, of postoperative complications in the time frame of this study. CONCLUSIONS: A postoperative serum PCT concentration of >0.5 ng/mL is highly suggestive of a postoperative complication. CRP changes do not contribute to predictive information. PMID- 16038390 TI - A potential propensity for failure secondary to clot embolism in neonatal ECMO. AB - OBJECTIVE: To report a single case of oxygenator failure caused by clot embolism originating from the bladder; and to discuss some preventative options. CASE REPORT: A 2.5 kg neonate with a diagnosis of influenza A received veno-arterial (V-A) extracorporeal membrane oxygenation (ECMO) for cardiorespiratory support. Halfway through treatment, she underwent an elective circuit change for numerous clots in her circuit. The patient continued to consume vast quantities of platelets and developed a fatal oxygenator failure after 18 days. DISCUSSION: Amongst the factors influencing the outcome in events of a sudden unexpected oxygenator failure are the severity of patient illness, the size of the clot relative to the size of the oxygenator, the availability of a previously primed circuit and the ease and speed of priming a new oxygenator. CONCLUSION: There is a need for improvement in the design of small oxygenators and ECMO circuits. Adjustment of the coagulation parameters and lowering the tolerance towards clots in the circuit by electively changing them may reduce the incidence of sudden unexpected oxygenator failure. However, using a slightly larger Medos oxygenator may gain valuable time needed to arrange an oxygenator/circuit change. PMID- 16038391 TI - Anoplocephala perfoliata of horses--significant scope for further research, improved diagnosis and control. AB - Anoplocephala perfoliata is the commonest tapeworm parasite of horses and is incriminated as a significant cause of clinical disease (e.g., ileocaecal intussusception, caeco-caecal intussusception and/or caecal perforation), particularly in horses chronically infected with large numbers of worms. The high prevalence (approximately 20-80%) of the parasite in some countries suggests an increased risk of clinical cases. In spite of research, there is still a paucity of information regarding the pathogenesis of the disease, the epidemiology of the parasite in different geographical regions and there are significant limitations with the diagnosis of infection. The present article provides an account of the biology, epidemiology and pathogenic effects of A. perfoliata, the diagnosis of infection and treatment. It highlights some gaps in knowledge of the parasite and the disease it causes, and suggests opportunities for future research and prospects for improved diagnosis, prevention and control. PMID- 16038392 TI - Diagnosing genetically diverse avian malarial infections using mixed-sequence analysis and TA-cloning. AB - Birds harbouring several malarial parasites are common in the wild, and resolving such multiple infections is important for our understanding of host-parasite relationships. We propose a simple and reasonably accurate method for detecting and resolving multiple infections, based on the analysis of parasite cytochrome b DNA sequences: genetically mixed infections are first identified by double nucleotide peaks on sequence electropherograms, and later retrieved by TA cloning. We applied this method to wild birds, and to experimentally created mixes with varying proportion of two parasites (Plasmodium spp. and Haemoproteus spp.). In general, the method was very efficient in detecting and resolving multiple infections, but some problems were encountered. Several multiple infections were erroneously scored as simple, either because one of the parasite lineages was a better target for the primers used, or because it was much more abundant in the mix. On the other hand, single nucleotide substitutions and template switching during PCR produced artificial sequences in some clones. We discuss the utility of the method, and propose a framework for its use when screening for genetically diverse avian malarial parasites. PMID- 16038393 TI - Molecular evolution and intragenic recombination of the merozoite surface protein MSP-3alpha from the malaria parasite Plasmodium vivax in Thailand. AB - The merozoite surface antigens of malaria parasites are prime anti morbidity/mortality vaccine candidates. However, their highly polymorphic nature requires extensive surveys of parasite populations to validate vaccine designs. Previous studies have found 3 molecular types (A, B and C) of the Plasmodium vivax merozoite surface protein 3a (PvMSP-3alpha) among parasite field populations. Here we analysed complete PvMSP-3alpha sequences from 17 clinical P. vivax isolates from Thailand and found that the nucleotide diversity was as high as that from samples widely separated by time and space. The polymorphic sites were not randomly distributed but concentrated in the N-terminal Ala-rich domain (block 2A), which is partially deleted in type B and C sequences. The size variations among type A sequences were due to small indels occurring in block 2A, whereas type B and C sequences were uniform in length with each type having a different large deletion. Analysis of synonymous and non-synonymous substitutions suggested that different selection forces were operating on different regions of the molecule. The numerous recombination sites detected within the Ala-rich domain suggested that intragenic recombination was at least partially responsible for the observed genetic diversity of the PvMSP-3alpha gene. Phylogenetic analysis failed to link any alleles to a specific geographical origin, even when different domains of PvMSP-3alpha were used for analysis. The highly polymorphic nature and lack of geographical clustering of isolates suggest that more systematic investigations of the PvMSP-3alpha gene are needed to explore its evolution and vaccine potential. PMID- 16038394 TI - The proteasome inhibitor MLN-273 blocks exoerythrocytic and erythrocytic development of Plasmodium parasites. AB - Protein degradation is regulated during the cell cycle of all eukaryotic cells and is mediated by the ubiquitin-proteasome pathway. Potent and specific peptide derived inhibitors of the 20S proteasome have been developed recently as anti cancer agents, based on their ability to induce apoptosis in rapidly dividing cells. Here, we tested a novel small molecule dipeptidyl boronic acid proteasome inhibitor, named MLN-273 on blood and liver stages of Plasmodium species, both of which undergo active replication, probably requiring extensive proteasome activity. The inhibitor blocked Plasmodium falciparum erythrocytic development at an early ring stage as well as P. berghei exoerythrocytic progression to schizonts. Importantly, neither uninfected erythrocytes nor hepatocytes were affected by the drug. MLN-273 caused an overall reduction in protein degradation in P. falciparum, as demonstrated by immunoblots using anti-ubiquitin antibodies to label ubiquitin-tagged protein conjugates. This led us to conclude that the target of the drug was the parasite proteasome. The fact that proteasome inhibitors are presently used as anti-cancer drugs in humans forms a solid basis for further development and makes them potentially attractive drugs also for malaria chemotherapy. PMID- 16038395 TI - Comparison of manual and homogenizer methods for preparation of tick-derived stabilates of Theileria parva: equivalence testing using an in vitro titration model. AB - Theileria parva sporozoite stabilates are used in the infection and treatment method of immunization, a widely accepted control option for East Coast fever in cattle. T. parva sporozoites are extracted from infected adult Rhipicephalus appendiculatus ticks either manually, using a pestle and a mortar, or by use of an electric homogenizer. A comparison of the two methods as a function of stabilate infectivity has never been documented. This study was designed to provide a quantitative comparison of stabilates produced by the two methods. The approach was to prepare batches of stabilate by both methods and then subject them to in vitro titration. Equivalence testing was then performed on the average effective doses (ED). The ratio of infective sporozoites yielded by the two methods was found to be 1.14 in favour of the manually ground stabilate with an upper limit of the 95% confidence interval equal to 1.3. We conclude that the choice of method rests more on costs, available infrastructure and standardization than on which method produces a richer sporozoite stabilate. PMID- 16038396 TI - Asymptomatic giardiasis and growth in young children; a longitudinal study in Salvador, Brazil. AB - This study sought to assess the effect of giardiasis on growth of young children. In Salvador, northeast Brazil, 597 children initially aged 6 to 45 months were followed for a year in 1998/9, measured anthropometrically thrice, every 6 months, and monitored for diarrhoea prevalence twice weekly. Stool samples were collected and examined during the second round of anthropometry, and infected children were treated 39 days later, on average (S.D. 20 days). For each 6-month interval, the gains in z-scores of infected and uninfected children were compared, after adjustment for potential confounding factors, including longitudinal prevalence of diarrhoea. No significant difference was found for the first interval but in the second, the gain in adjusted height-for-age z-score was 0.09 less in infected than uninfected children, equivalent to a difference in height gain of 0.5 cm. The shortfall in growth was greater in children who remained free of diarrhoea, and was significantly correlated with the proportion of the second interval during which the child had remained untreated. We conclude that Giardia can impede child growth even when asymptomatic, presumably through malabsorption. This finding challenges the view that young children found to have asymptomatic giardiasis in developing countries should not be treated. PMID- 16038397 TI - Mode of action of a potentially important excretory--secretory product from Giardia lamblia in mice enterocytes. AB - Giardia, a common enteric protozoan parasite is a well-recognized cause of diarrhoeal illness. The detailed mechanism of diarrhoea due to this infection is not well understood. A 58 kDa enterotoxin (ESP) was purified from the excretory secretory product of the parasite. The present study was designed to investigate the mode of action of this enterotoxin of G. lamblia in mice enterocytes. An increase in cyclic adenosine monophosphate level, as well as intracellular Ca2+ concentration, was observed in the ESP-triggered enterocytes. The levels of phospholipase Cgamma1 and inositol triphosphate were found to be upregulated. The activity of protein kinase C (PKC) in the enterocytes was also enhanced following stimulation with the ESP. An increase in the level of reactive oxygen species in ESP-stimulated cells correlated well with the decline in the activity of antioxidant enzymes (superoxide dismutase and catalase). The significantly high levels of nitrite and citrulline indicated the generation of reactive nitrogen intermediates in the ESP-triggered enterocytes. Thus, ESP could induce cross-talk among the different signal transduction pathways in the enterocytes, which could together bring about a common secretory response. PMID- 16038398 TI - Heterogeneity in extracellular nucleotide hydrolysis among clinical isolates of Trichomonas vaginalis. AB - Trichomonas vaginalis is a parasitic protozoan that causes trichomonosis, a sexually-transmitted disease, with serious sequelae to women and men. As the host parasite relationship is complex, it is important to investigate biochemical aspects of the parasite that contribute to our understanding of trichomonal biology and pathogenesis. Nucleoside triphosphate diphosphohydrolase 1 (NTPDase 1), which hydrolyses extracellular ATP and ADP, and ecto-5'-nucleotidase, which hyrolyses AMP, have been characterized in laboratory isolates of T. vaginalis. Here we show that the extracellular ATP: ADP hydrolysis ratio varies among fresh clinical isolates, which presented higher ATPase and ADPase activities than long term-grown isolates. Growth of parasites in iron-replete and iron-depleted medium resulted in different, albeit minor, patterns in extracellular ATP and ADP hydrolysis among isolates. Importantly, some isolates had low or absent ecto-5' nucleotidase activity, regardless of environmental conditions tested. For isolates with ecto-5'-nucleotidase activity, high- and low-iron trichomonads had increased and decreased levels of activity, respectively, compared to organisms grown in normal TYM-serum medium. This suggests a regulation in expression of either the enzyme amounts and/or activity under the control of iron. Finally, we found no correlation between the presence or absence of dsRNA virus infection among trichomonad isolates and NTPDase and ecto-5'-nucleotidase activities. PMID- 16038399 TI - Paleohaemoproteus burmacis gen. n., sp. n. (Haemospororida: Plasmodiidae) from an Early Cretaceous biting midge (Diptera: Ceratopogonidae). AB - Paleohaemoproteus burmacis gen. n., sp. n. (Haemospororida: Plasmodiidae) is described from the abdominal cavity of a female biting midge (Diptera: Ceratopogonidae) preserved in 100 million year old amber from Myanmar (Burma). The description is based on the developmental stages of oocysts and sporozoites. The fossil species differs from extant species of Haemoproteus by its wide range of oocyst sizes, small sporozoites and occurrence in an extinct species of biting midge. Numerous sporozoites in the abdominal cavity suggest that the biting midge was an effective vector of this malarial parasite. Characters of the biting midge suggest that the host was a large, cold-blooded vertebrate. This is the earliest record of a malaria parasite and first indication that Early Cretaceous reptiles were infected with haemosporidial parasites. PMID- 16038400 TI - Characterization of an extracellular serine protease of Leishmania (Leishmania) amazonensis. AB - A serine protease was purified 942-fold from culture supernatant of L. amazonensis promastigotes using (NH4)2SO4 precipitation followed by affinity chromatography on aprotinin-agarose and continuous elution electrophoresis by Prep Cell, yielding a total recovery of 61%. The molecular mass of the active enzyme estimated by SDS-PAGE under conditions of reduction was 56 kDa and 115 kDa under conditions of non-reduction, suggesting that the protease is a dimeric protein. Additionally, it was found to be a non-glycosylated enzyme, with a pI of 5.0. The optimal pH and temperature of the enzyme were 7.5 and 28 degrees C respectively, using alpha-N-rho-tosyl-L-arginine-methyl ester (L-TAME) as substrate. Assays of thermal stability indicated that 61% of the enzyme activity was preserved after 1 h of pre-treatment at 42 degrees C. Haemoglobin, bovine serum albumin (BSA), ovalbumin, fibrinogen, collagen, gelatin and peptide substrates containing arginine in an ester bond and amide substrates containing hydrophobic residues at the P1 site were hydrolysed by this extracellular protease. The insulin beta-chain was also hydrolysed by the enzyme and many peptidic bonds were susceptible to the protease action, and 4 of them (L11-V12, E3-A14, L15-Y16 and Y16-L17) were identified. Inhibition studies suggested that the enzyme belongs to the serine protease class inhibited by calcium and manganese and activated by zinc. These findings show that this enzyme of L. amazonensis is a novel serine protease, which differs from all known flagellate proteases characterized. PMID- 16038401 TI - Spatial risk prediction and mapping of Schistosoma mansoni infections among schoolchildren living in western Cote d'Ivoire. AB - The objectives of this study were (1) to examine risk factors for Schistosoma mansoni infection among schoolchildren living in western Cote d'Ivoire, and (2) to carry forward spatial risk prediction and mapping at non-sampled locations. First, demographic and socio-economic data were obtained from 3818 children, aged 6-16 years, from 55 schools. Second, a single stool sample was examined from each child by the Kato-Katz technique to assess infection status of S. mansoni and its intensity. Third, remotely sensed environmental data were derived from satellite imagery and digitized ground maps. With these databases a comprehensive geographical information system was established. Bayesian variogram models were applied for spatial risk modelling and prediction. The infection prevalence of S. mansoni was 38.9%, ranging from 0% to 89.3% among schools. Results showed that age, sex, the richest wealth quintile, elevation and rainfall explained the geographical variation of the school prevalences of S. mansoni infection. The goodness of fit of different spatial models revealed that age, sex and socio economic status had a stronger influence on infection prevalence than environmental covariates. The generated risk map can be used by decision-makers for the design and implementation of schistosomiasis control in this setting. If successfully validated elsewhere, this approach can guide control programmes quite generally. PMID- 16038402 TI - Differential infectivity and immunopathology in murine experimental infections by two natural clones belonging to the Trypanosoma cruzi I lineage. AB - Immunopathology of Chagas' disease in Balb/c mice infected with 2 Trypanosoma cruzi clones, belonging to the T. cruzi I lineage and presenting different in vitro virulence (P/209 cl1 > SO34 c14) was compared. In the acute phase, evading mechanisms such as parasite-induced lymphocyte polyclonal activation and T cell immunosuppression were higher in mice infected with the clone giving a higher parasitaemia (P/209 cl1). A similar increase of non-specific isotypes was observed in both infections with IgG2a prevalence. Interestingly, CD8+ cell hypercellularity and lymphocyte immunosuppression were observed during the chronic phase (245 days post-infection) in mice infected by the most virulent clone. In the same way, the parasite-specific antibody response was more intense in P/209 cl1-infected mice over the acute phase. During the chronic phase this response remarkably dropped down in SO34 cl4-infected mice exclusively. Finally, P/209 cl1-infected mice presented a more severe inflammation and tissue damage in heart and quadriceps than SO34 cl4-infected mice. This comparative study showed differences between the two clones: a higher virulence in vivo being clearly associated with a greater ability to induce evasion mechanisms and severe tissue damage. PMID- 16038403 TI - Density dependence and overdispersion in the transmission of helminth parasites. AB - The influence of density-dependent processes on the transmission of parasitic helminths is determined by both the severity of the regulatory constraints and the degree of parasite overdispersion among the host population. We investigate how overdispersed parasite distributions among humans influence transmission levels in both directly- and indirectly-transmitted nematodes (Ascaris lumbricoides and Onchocerca volvulus). While past work has assumed, for simplicity, that density dependence acts on the average worm load, here we model density-dependence as acting on individual parasite burdens before averaging across hosts. A composite parameter, which we call the effective transmission contribution, is devised to measure the number of transmission stages contributed by a given worm burden after incorporating over-dispersion in adult worm mating probabilities and other density-dependent mechanisms. Results indicate that the more overdispersed the parasite population, the greater the effect of density dependence upon its transmission dynamics. Strong regulation and parasite overdispersion make the relationship between mean worm burden and its effective contribution to transmission highly non-linear. Consequently, lowering the intensity of infection in a host population using chemotherapy may produce only a small decline in transmission (relative to its initial endemic level). Our analysis indicates that when parasite burden is low, intermediate levels of parasite clustering maximize transmission. Implications are discussed in relation to existing control programmes and the spread of anthelmintic resistance. PMID- 16038404 TI - Transmission dynamics of Echinococcus multilocularis; its reproduction number, persistence in an area of low rodent prevalence, and effectiveness of control. AB - On the basis of high prevalences of Echinococcus multilocularis in the growing fox populations in Central Europe, its total biomass may have increased significantly in the past 20 years. E. multilocularis is now also found in areas outside the known endemic area in Central Europe. Therefore, E. multilocularis, the causative agent of a serious parasitic zoonosis, might be of major concern for public health and a challenge to control. Some experimental field trials to control E. multilocularis using an anti-worm drug reduced parasite burden in a contaminated region during the control campaign, but failed to eradicate the parasite completely. It was our aim to develop a mathematical model describing the biomass of egg, larval, and adult worm stages of the E. multilocularis life cycle, and simulate a hypothetical control campaign. Additionally, we derived the reproduction number of this parasite and explored conditions for the persistence of the parasite's life-cycle. Our model shows that while control campaigns rapidly reduce the worm burden in the definitive host, and consequently eggs in the environment, the pool of larvae in the intermediate host remains large. The parasite's life-cycle persists in a region where prevalence in the intermediate host is low (approximately 1%). Therefore, we conclude that the parasite is likely to re-emerge if control is discontinued on the basis of reduced worm population. Continued treatment of the definitive host is required to eradicate the larval stage of the parasite from the intermediate host population. PMID- 16038405 TI - Molecular cloning and characterization of Cpn60 in the free-living nematode Plectus acuminatus. AB - Heat shock proteins (Hsps) have provoked interest not only because of their involvement in human diseases but also for their potential as biomarkers of environmental pollution. Whereas the former interest is covered by numerous reports, the latter is an exciting new field of research. We report the isolation of the full-length cpn60 messenger ribonucleic acid (mRNA) and partial genomic deoxyribonucleic acid from the free-living, environmental sentinel nematode Plectus acuminatus, a species used in classical ecotoxicity tests. Although the primary sequence displays high identity scores to other nematodes and human Cpn60 (75% and 70%, respectively), the intron-exon structure differs markedly. Furthermore, although mRNA levels remained constant after exposure to ZnCl2 (0 330 microM) under laboratory conditions, protein levels increased significantly in a dose-dependent manner. In conclusion, this first account of molecular genetic similarities and differences of Cpn60 in a neglected nematode taxon provides a valuable insight into its potential uses in gene-based ecotoxicological risk assessment exercises. PMID- 16038407 TI - Temperature differentially affects adenosine triphosphatase activity in Hsc70 orthologs from Antarctic and New Zealand notothenioid fishes. AB - To test the temperature sensitivity of molecular chaperones in poikilothermic animals, we purified the molecular chaperone Hsc70 from 2 closely related notothenioid fishes--the Antarctic species Trematomus bernacchii and the temperate New Zealand species Notothenia angustata--and characterized the effect of temperature on Hsc70 adenosine triphosphatase (ATPase) activity. Hsc70 ATPase activity was measured using [alpha-32P]-adenosine triphosphate (ATP)-based in vitro assays followed by separation of adenylates by thin-layer chromatography. For both species, a significant increase in Hsc70 ATPase activity was observed across a range of temperatures that was ecologically relevant for each respective species. Hsc70 from T bernacchii hydrolyzed 2-fold more ATP than did N angustata Hsc70 at 0 degrees C, suggesting that the Antarctic molecular chaperone may be adapted to function more efficiently at extreme cold temperatures. In addition, Q10 measurements indicate differential temperature sensitivity of the ATPase activity of Hsc70 from these differentially adapted fish that correlates with the temperature niche inhabited by each species. Hsc70 from T bernacchii was relatively temperature insensitive, as indicated by Q10 values calculated near 1.0 across each temperature range measured. In the case of Hsc70 purified from N angustata, Q10 values indicated thermal sensitivity across the temperature range of 0 degrees C to 10 degrees C, with a Q10 of 2.714. However, Hsc70 from both T bernacchii and N angustata exhibited unusually high thermal stabilities with ATPase activity at temperatures that far exceeded temperatures encountered by these fish in nature. Overall, as evidenced by in vitro ATP hydrolysis, Hsc70 from T bernacchii and N angustata displayed biochemical characteristics that were supportive of molecular chaperone function at ecologically relevant temperatures. PMID- 16038406 TI - Heat shock proteins in cancer: diagnostic, prognostic, predictive, and treatment implications. AB - Heat shock proteins (Hsps) are overexpressed in a wide range of human cancers and are implicated in tumor cell proliferation, differentiation, invasion, metastasis, death, and recognition by the immune system. We review the current status of the role of Hsp expression in cancer with special emphasis on the clinical setting. Although Hsp levels are not informative at the diagnostic level, they are useful biomarkers for carcinogenesis in some tissues and signal the degree of differentiation and the aggressiveness of some cancers. In addition, the circulating levels of Hsp and anti-Hsp antibodies in cancer patients may be useful in tumor diagnosis. Furthermore, several Hsp are implicated with the prognosis of specific cancers, most notably Hsp27, whose expression is associated with poor prognosis in gastric, liver, and prostate carcinoma, and osteosarcomas, and Hsp70, which is correlated with poor prognosis in breast, endometrial, uterine cervical, and bladder carcinomas. Increased Hsp expression may also predict the response to some anticancer treatments. For example, Hsp27 and Hsp70 are implicated in resistance to chemotherapy in breast cancer, Hsp27 predicts a poor response to chemotherapy in leukemia patients, whereas Hsp70 expression predicts a better response to chemotherapy in osteosarcomas. Implication of Hsp in tumor progression and response to therapy has led to its successful targeting in therapy by 2 main strategies, including: (1) pharmacological modification of Hsp expression or molecular chaperone activity and (2) use of Hsps in anticancer vaccines, exploiting their ability to act as immunological adjuvants. In conclusion, the present times are of importance for the field of Hsps in cancer, with great contributions to both basic and clinical cancer research. PMID- 16038408 TI - DNA sequence determinants of nuclear protein binding to the c-Ha-ras antioxidant/electrophile response element in vascular smooth muscle cells: identification of Nrf2 and heat shock protein 90 beta as heterocomplex components. AB - The antioxidant/electrophile response element (ARE/EpRE) is a cis-acting element involved in redox regulation of c-Ha-ras gene. Protein binding to the ARE/EpRE may be credited to deoxyribonucleic acid sequence; therefore, studies were conducted to evaluate the influence of internal and flanking regions to the 10-bp human c-Ha-ras ARE/EpRE core (hHaras10) on nuclear protein binding in oxidant treated vascular smooth muscle cells. A protein doublet bound to an extended oligonucleotide comprising the ARE/EpRE core in genomic context (hHaras27), whereas a single complex bound to hHarasl0. Protein binding involved specific interactions of 25- and 23-kDa proteins with hHarasl0, and binding of 80-, 65-, and 55-kDa proteins to hHaras27. Competition assays with hNQO1 and rGSTA2 confirmed the specificity of deoxyribonucleic acid-protein interactions and indicated preferred binding of p25 and p23 to the c-Ha-ras ARE/EpRE. "NNN" sequences within the core afforded unique protein-binding profiles to the c-Ha ras ARE/EpRE. In addition, Nrf2 and heat shock protein 90beta (p80) were identified as components of the c-Ha-ras ARE/EpRE heterocomplex. We conclude that both internal bases and flanking sequences regulate nuclear protein recruitment and complex assembly on the c-Ha-ras ARE/EpRE. PMID- 16038409 TI - Plasma antibodies to heat shock protein 60 and heat shock protein 70 are associated with increased risk of electrocardiograph abnormalities in automobile workers exposed to noise. AB - In the living and working environment, stressful factors, such as noise, can cause health problems including cardiovascular diseases and noise-induced hearing loss. Some heat shock proteins (Hsps) play an important role in protecting cardiac cells against ischemic injury, and antibodies against these Hsps are associated with the development and prognosis of atherogenesis, coronary heart disease, and hypertension. Whether the presence of such antibodies is associated with abnormal electrocardiography (ECG) in stressed autoworkers exposed to chronic noise is presently unknown. Therefore, we investigated the association between the levels of plasma anti-Hsp60 and anti-Hsp70 with electrocardiograph abnormality in 396 autoworkers exposed to different noise levels by using Western blot, ECG, and multivariate logistic regression analysis. The results showed that the increase in levels of anti-Hsp70 was associated with a higher risk of ECG abnormalities characteristic of chronic myocardial ischemia (P < 0.05), conductive abnormality (P < 0.01), or heart displacement (P < 0.05); in contrast, elevated anti-Hsp60 was related to ECG abnormalities characteristic of sinus arrhythmia, chronic myocardial ischemia, and ectopic rhythm (P < 0.01 for all). Overall, high levels of both anti-Hsp70 and anti-Hsp60 were associated with significantly increased risk of ECG abnormalities (odds ratio [OR] = 1.73 and 95% confidence interval [Cl] = 1.04-2.86 for anti-Hsp70 and OR = 1.36 and 95% Cl = 1.07-1.72 for anti-Hsp60) with and without adjustment for cumulative noise exposure (OR = 1.96 and 95% Cl = 1.20-3.21 for anti-Hsp70 and OR = 3.93 and 95% Cl = 1.72-8.92 for anti-Hsp60). These findings suggest that the production of both anti-Hsp70 and anti-Hsp60 may be independent risk factors for the development and progression of abnormal ECG and therefore possibly cardiovascular diseases in autoworkers exposed to occupational noise. PMID- 16038410 TI - Retinoid- and sodium-butyrate-induced decrease in heat shock protein 70 membrane positive tumor cells is associated with reduced sensitivity to natural killer cell lysis, growth delay, and altered growth morphology. AB - Human tumors frequently present heat shock protein 70 (Hsp70) on their cell membranes, whereas corresponding normal tissues fail to do so. Therefore, an Hsp70 membrane-positive phenotype provided a tumor-specific marker. Moreover, membrane-bound Hsp70 provides a target structure for the cytolytic attack mediated by natural killer (NK) cells. Vitamin A derivatives 13-cis retinoic acid (13-RA) and all-trans retinoic acid (ATRA) and sodium-butyrate (SBU) are known for their redifferentiating capacity. Therefore, we asked the question whether loss in tumorigenicity might be associated with a reduced Hsp70 membrane expression. For our studies we used epithelial colon (CX+/CX-) and thyroid (ML-1) cancer cells, with initially different Hsp70 cell surface expression pattern. After treatment up to 7 weeks with freshly prepared 13-RA, ATRA, and SBU at nonlethal concentrations of 10 microM, 1 microM, and 0.5 mM, respectively, growth morphology, Hsp70 levels, and sensitivity toward Hsp70-specific NK cells were compared with that of untreated tumor cells. Significant growth delay was determined in CX+ tumor cells after 6 weeks treatment with 13-RA. Concomitantly, growth morphology changed from spheroid cell clusters to monolayers. Despite a weak increase in cytosolic Hsp70, the percentage of Hsp70 membrane-positive cells dropped significantly after repeated treatments with 13-RA and ATRA in CX+ and ML 1 but not in CX- tumor cells. Similar results were observed with SBU. Functionally, the decrease in Hsp70 membrane-positive CX+ and ML-1 cells correlated with a reduced sensitivity to lysis mediated by NK cells. In summary, redifferentiating agents predominantly affected Hsp70 membrane-positive tumors. The decrease in Hsp70 membrane positivity correlated with a lower sensitivity to NK lysis, growth delay, and altered growth morphology. PMID- 16038411 TI - C-terminus of heat shock protein 70-interacting protein facilitates degradation of apoptosis signal-regulating kinase 1 and inhibits apoptosis signal-regulating kinase 1-dependent apoptosis. AB - Apoptosis signal-regulating kinase 1 (ASK1) is a mitogen-activated protein kinase kinase kinase (MAPKKK) that is regulated under conditions of cellular stress. ASK1 phosphorylates c-Jun N-terminal kinase (JNK) and elicits an apoptotic response. ASK1 activity is regulated at multiple levels, 1 of which is through inhibition by cytosolic chaperones of the heat shock protein (Hsp) 70 family. Among the proteins that determine Hsp70 function, CHIP (C-terminus of Hsp70 interacting protein) is a cochaperone and ubiquitin ligase that interacts with Hsp70 through an amino-terminal tetratricopeptide repeat (TPR) domain. Prominent among the cellular functions mediated by CHIP is protection against physiologic stress. Because ASK1 is known to contain a TPR-acceptor site, we examined the role of CHIP in regulating ASK1 function. CHIP interacted with ASK1 in a TPR dependent fashion and induced ubiquitylation and proteasome-dependent degradation of ASK1. Targeting of ASK1 by CHIP inhibited JNK activation in response to oxidative challenge and reduced ASK1-dependent apoptosis, whereas short interfering RNA (siRNA)-dependent depletion of CHIP enhanced JNK activation. Consistent with its ability to reduce cytoplasmic ASK1 levels, CHIP triggered the translocation of ASK1 partner protein death-associated protein (Daxx) into the nucleus, where it is known to activate an antiapoptotic response. These results indicate that CHIP regulates ASK1 activity by inducing its ubiquitylation and degradation, which, together with its effects on Daxx localization, provides a mechanism for the antiapoptotic effects of CHIP observed in the face of cellular and physiologic stress. PMID- 16038413 TI - [Prevalence and characteristics of different types of homeboundness among community-living older adults]. AB - BACKGROUND: Little is known about the epidemiologic features of different types of homeboundness among the elderly. PURPOSE: This cross-sectional study examined prevalence and characteristics of "type 1" and "type 2" homeboundness (see definitions below) among community-living older adults. METHODS: The subjects comprised all residents aged 65 years and over living in Yoita, Niigata Prefecture, and Hatoyama, Saitama Prefecture. Subject data on sociodemographics, and physical, mental and social functioning were collected through in-person interview. Persons were defined as being homebound if he/she went outdoors only once a week or less often. Homeboundness was further classified into "type 1" or "type 2", based on the hierarchical mobility level classification (levels 1 or 2 vs. levels 3, 4, or 5). "Type 1" homebound persons included those who could not get out into the neighborhood without assistance (i.e, levels 3, 4, or 5). "Type 2" included those who were homebound, though they could get out at least into the neighborhood unassisted (i.e., levels 1 or 2). We focused on characteristics of "type 1" and "type 2" homeboundness as compared with those of respective controls, ie., non-homebound persons within the same mobility categories. RESULTS: Out of the eligible subjects (1588 in Yoita, and 1135 in Hatoyama), 1544 and 1002 persons participated in the survey (response rates of 97.2% and 88.3%, respectively). Among the participants, "type 1" and "type 2" homeboundness was found for 4.1% and 5.4%, respectively, in Yoita, and 3.3% and 6.8% in Hatoyama. After adjustment for potential confounders such as age, gender and mobility level, we found a significant regional difference in the prevalence of "type 2" but not of "type 1" (OR of "type 2" for Hatoyama/Yoita 1.44; 95% CI 1.02-2.03). Both types of homeboundness increased with advancing age; "type 1" and "type 2" featured in over 10% of persons aged at least 85 years and 80 years, respectively. Even after controlling for potential confounders, "type 2" showed a higher prevalence with walking disability and incontinence, and reported lower self-rated health, more depressed mood, lower functional capacity and lower social functioning. "Type 1" showed a higher prevalence with fear of falls, but a lower prevalence with basic ADL disability and a high score for Intellectual Activity, indicating reduced self-efficacy. CONCLUSIONS: Prevalence of "type 1" and "type 2" homeboundness among community-living older adults differs depending on the residential area and age of the subjects. A substantial proportion of "type 2" homebound persons are at high risk of functional decline, indicating that "type 2" as well as "type 1" homebound persons need care-preventive programs. PMID- 16038412 TI - Structural instability caused by a mutation at a conserved arginine in the alpha crystallin domain of Chinese hamster heat shock protein 27. AB - Mutations in the alpha-crystallin domain of 4 of the small heat shock proteins (sHsp) (Hsp27/HspB1, alphaA-crystallin/ HspB4, alphaB-crystallin/HspB5, and HspB8) are responsible for dominant inherited diseases in humans. One such mutation at a highly conserved arginine residue was shown to cause major conformational defects and intracellular aggregation of alphaA- and alphaB crystallins and HspB8. Here, we studied the effect of this Arg mutation on the structure and function of Hsp27. Chinese hamster Hsp27 with Arg148 replaced by Gly (Hsp27R148G) formed dimers in vitro and in vivo, which contrasted with the 12 or 24-subunit oligomers formed by the wild-type protein (Hsp27WT). Despite these alterations, Hsp27R148G had a chaperone activity almost as high as Hsp27WT. The dimers of Hsp27R148G did not further deoligomerize on phosphorylation and like the dimers formed by phosphorylated Hsp27WT were not affected by the deletion of the N-terminal WD/EPF (single letter amino acid code) motif, suggesting that mutation of Arg148, deletion of the N-terminal WD/EPF motif, and phosphorylation of Ser90 may produce similar structural perturbations. Nevertheless, the structure of Hsp27R148G appeared unstable, and the mutated protein accumulated as aggregates in many cells. Both a lower basal level of phosphorylation of Hsp27R148G and the coexpression of Hsp27WT could reduce the frequency of formation of these aggregates, suggesting possible mechanisms regulating the onset of the sHsp-mediated inherited diseases. PMID- 16038414 TI - [Social relationaship factors and risk of care requirement in Japanese elderly]. AB - OBJECTIVE: To identify associations between social relationship factors and risk of requirement for care in elderly people living at home. METHOD: In February 2000, a questionnaire was distributed to all residents aged 65 and over in a city in Aichi prefecture, Japan. The number of responses was 3596, a return rate of 72.0%. The records of 2725 subjects who were not under care as of April 2000 were selected as a sample and treated with Cox's proportional hazards model in order to analyze associations between social relationship variables and the risk of care requirement. RESULTS: (1) For males, the higher the acceptance of support, the higher the risk of care requirement (hazard ratio (HR) 1.12, 95% confidence interval (CI) 1.04-1.21). For females, the lower the contact frequency with separately-living family members, the higher the incidence (HR2.14, 95% CI 1.27 3.62). Also, the analysis of female subjects showed a strong association between not providing social support to others and the risk of care requirement (HR2.75, 95% CI 1.61-4.71). (2) Analysis of the effects of social support indicated that the male subjects who did not provide support to others but received such support themselves were at significantly high risk of care requirement (HR2.90, 95% CI 1.17-7.18). (3) In the case of female subjects who lived alone, unlike those with other living arrangements, there appeared to be no significant associations between involvement in social networks and their support provision to others, and the risk of care requirement. CONCLUSIONS: The study confirmed that gender and living arrangements both influence associations between the risk of care requirement in the elderly and social networks and support. The study also indicated that provision of support by the elderly is one important factor that can lower the risk of needing care. PMID- 16038415 TI - [Utility of cycle ergometer stress testing as a measure to assess health status and physical functioning for elderly people]. AB - PURPOSE: This study was carried out to determine the prevalence of abnormal cardiovascular findings for elderly persons on exercise stress testing, and examine the relationship between exercise capacity estimated by the test and other physical functions or self-rated functional capacity cross-sectionally. METHODS: Participants of this study were 83 elderly persons (24 males and 59 females, mean age, 80 years old) who were planning to undergo a falls prevention program. A questionnaire asking about functional capacity, physical functional testing (maximum stride length, maximum speed walking for 10 meters, one leg standing time, and grip strength), and cycle ergometer exercise stress testing were applied. The initial work load of exercise testing was 15 watts, and this was progressively increased by 15 watts every 3 minutes. Exercise capacity was estimated with reference to working time and subjects were divided into tertiles for each gender. Comparisons of physical functional testing and self-rated functional capacity among the groups were then performed. RESULTS: Of 83 participants, 65 subjects (78.3%) could complete the exercise stress testing. Next to leg fatigue (46%), blood pressure elevation (26.2%) was the most common reason for termination of the test. No evidence of ischemia was found in any subject during exercise stress testing. The values for walking speed and walking pitch were highest among male participants in the third (uppermost) tertile group with statistical significance. In addition, the value for one foot standing time was highest among female participants in the third tertile group, again with statistical significance. Regarding self-rated functional capacity, the rates for subjects who reported being able to rise from a seat in a vehicle, jump over a puddle, and step on an escalator were highest among female participants in the third tertile group, with statistical significance, whereas no significant differences were seen among male participants. No one suffered falls in the previous one year in either gender in the third tertile group. CONCLUSION: As the major finding, higher exercise capacity mainly for lower-extremity work, was associated with better physical functions and self-rated functional capacity. PMID- 16038416 TI - [A study on the factors related to smoking behavior among university students]. PMID- 16038417 TI - [Needs assessment for persons with hepatitis C virus (HCV) infection in a community in Osaka, 1990's]. PMID- 16038418 TI - [The influence of alcohol drinking on medical care expenditures among National Health Insurance beneficiaries]. AB - Heavy alcohol drinking is an important risk factor for some lifestyle-related diseases. However, the relationship between drinking habits and medical care expenditures has not been elucidated in a Japanese general population. We examined this relationship in a 9-year cohort study. Study participants were 2039 National Health Insurance beneficiaries aged 40-69 years living in a rural Japanese community. The baseline survey, with medical check-up, was performed in 1989 or 1990 and the subjects' drinking habits were estimated by questionnaire. Medical care use and costs were monitored by linkage with the National Health Insurance claim history files at baseline survey through the end of March 2000. Men who were daily heavy drinkers (ethanol intake > or =69 g per day) had significantly higher medical costs compared to participants who drank less, within 5 years after baseline. This difference was 2000 yen per month after adjusting for age, body mass index, smoking, systolic blood pressure, glycosuria, serum total cholesterol and alanine aminotransferase. Daily heavy male drinkers also had a higher hazard ratio for total mortality compared to occasional drinkers, by the Cox proportional hazard model (hazard ratio: 1.81), although it did not reach statistical significance (P = 0.153). In women, medical costs were almost the same for occasional and daily drinkers. Total medical costs among Japanese men aged 20 years and over may be decreased by about 8400 billon yen per year if we assume a 20% decrease in daily heavy drinkers. To reduce national medical costs, it is important to promote a program for heavy drinkers, as described in the "Health Japan 21" initiative, the Japanese national health promotion guidelines for the first 10 years of the 21st century. PMID- 16038419 TI - [The treatment cost at different stages of alcohol dependence in Japan]. AB - The treatment costs of alcohol dependence were calculated by disease stage and by method of treatment. Alcohol-related problems, mainly alcohol dependence, can be divided into four stages for convenience: (1) abnormal values found in health examinations, (2) preventive treatment as an outpatient or short-term hospitalization for education and testing, (3) basic outpatient treatment, and (4) inpatient treatment. The following results were obtained. (1) One example of expenses required for a routine examination was 936 points when converted to health insurance points. (2) The number of points for outpatient group psychotherapy in the first education stage and five additional examinations was 5175 points (6255 points with a designated mental health physician). The number of points for the program of medical tests and initial education in one week of hospitalization was 10,787. (3) For alcoholics, continued outpatient visits for two years, the number of points for only prescription of a drug to inhibit drinking such as disulfiram with no testing was 14,790 (19,650 points with a designated mental health physician). When the patient participated in alcohol daycare, the total number of points for three months of daycare with one meal each time was 46,020. (4) In cases of three-month inpatient treatment in our hospital including ordinary medical testing, the number of points for the first month for one patient was 57,280 points, plus 35,000 points each month in the 2nd and 3rd months for a total of 127,280 points. However, the total in our hospital is an average of 160,200 points when drugs and additional tests are included. As expected, the expenses required at the primary prevention stage were low, while those required for treatment after onset were high. Placing stress on prevention, providing proper treatment for patients and reducing relapses will reduce the burden including social expenses. PMID- 16038420 TI - [Economic aspect of alcohol-related disabilities in patients of gastroenterology and hepatology]. AB - Alcoholic patients frequently visit a medical center with the alcohol-related organ damage. The medical utilization and expenses of these patients is not only for the treatment of alcohol dependence, but also mainly for the treatment of alcohol-related organ diseases. This study was conducted to clarify the influence of alcohol dependence on the medical expense of internal medicine, especially of gastroenterology and hepatology. Forty four patients who visited the department of gastroenterology and hepatology of our institute since November 2001 were included in the study. All of the patients were heavy drinkers and were suffered from alcohol damages in various organs. Medical expenses of these patients were calculated from the amount of the claim for health insurance system record of our hospital. Average cumulative number of the patients per month was 15.5 +/- 2.8 in out patient and 4.2 +/- 2.4 in hospitalized patient. Medical expenses in these patients, however, were lower in out patients than in hospitalized patients. Cumulative medical expense per month was 238,000 +/- 66,000 yen and 1,801,000 +/- 1,338,000 yen respectively. Alcoholic liver cirrhosis is one of the most frequently encountered disorder related to heavy drinking and needs hospitalization for the treatment of complications of this end stage liver disease. 13 patients (29.5%) were diagnosed as having alcoholic liver cirrhosis in this study. Some of the patients successfully abstained, but irreversible liver damage necessitated repeated hospitalization for the treatment of decompensated cirrhosis. As a result, cumulative medical expense per patient with alcoholic liver cirrhosis was significantly larger than those with noncirrhotic patient. These results suggest that abstinence from alcohol in early stage of alcoholic liver disease, before development of cirrhosis, is important from economic point of view. PMID- 16038421 TI - [Analysis of medical care costs for gastroenterological inpatients with alcohol related disease]. AB - Current estimates suggest that approximately 2.3-2.5 million people in Japan are alcoholics. Of these patients, less than 1% visit alcohol outpatient clinics; most patients visit general clinics. Alcohol is associated with disorders of the gastrointestinal system, circulatory system, nervous system, and other organs. The costs for medical care impose a heavy burden on healthcare financing. Among all, the costs for alcohol-related gastrointestinal diseases are enormous. Reports show that the percentages of medical care costs for alcohol-related gastrointestinal diseases is as high as about 29% of the costs for all gastrointestinal diseases. Our analysis has found that hospital and treatment costs for alcohol-related liver and pancreatic diseases amounted to 35.2% of the costs for all liver and pancreatic diseases. Furthermore, results indicated that the average daily hospital and treatment costs for patients with alcohol-related liver diseases were significantly higher than the costs for patients with non alcohol-related liver diseases. To reduce medical care costs for alcohol-related diseases, not only treatment of such diseases but also preventive care for pre alcoholics is crucial. In this context, close contact between general clinics and alcohol outpatient clinics are important, and a network system of support for patients with alcohol-related diseases should be established. PMID- 16038422 TI - [Functional significance of L-type high voltage-gated calcium channels after sustained ethanol exposure]. AB - Functional significance of enhancement of L-type high voltage-gated calcium channels (HVCCs) induced by sustained ethanol exposure in mouse cerebral cortical neurons were focused in this review. Sustained ethanol exposure to the neurons induced up-regulation of L-type HVCCs, which was due to increased expression of a 1 and a 2/ delta 1 subunits. In the cerebral cortex prepared from alcohol dependent mice, similar pattern of alterations in the expressed subunits examined were also found. These data indicate that the increased expression of alpha 1 and alpha 2/ delta 1 subunits of L-type HVCCs is an essential biochemical event in cerebral cortex of alcohol-dependent mouse. PMID- 16038423 TI - [Serine protease inhibitors prevent alcoholic liver injury]. AB - The hepatotoxic effects of alcohol have been described in detail, but factors responsible for its hepatotoxicity have only partially characterized. It now appears that Kupffer cell derived TNF-alpha participates in several aspects of alcoholic liver injury. On the other hand, protease inhibitors have been used successfully for treatment of intractable diseases in which TNF-alpha is involved in the pathogenesis, including ulcerative colitis and Crohn's disease. Here, we will review new evidence for the proposal that serine protease inhibitors prevents alcoholic liver injury via mechanisms dependent on Kupffer cell derived TNF-alpha. PMID- 16038424 TI - [Fetal alcohol spectrum disorders (FASD) among Japanese children of alcoholic mothers]. AB - OBJECTIVE: This study was carried out to examine fetal alcohol spectrum disorders (FASD) among Japanese children of alcoholic mothers. This is the first report concerning FASD in Japan. METHODS: The subjects were 30 alcoholic women who were inpatients in the Kurihama Alcoholism Center and had given birth to children. They were subjected to a semi-structured interview by the author. Sixty healthy women who had not drunk during pregnancy were used as a control group, and they also underwent semi-structured interviews. The alcoholic women were divided into two groups, 13 who drunk during pregnancies and 17 who did not drink. Twenty children experienced of prenatal alcohol exposure and 40 children did not. The three groups; i.e., 13 alcoholic mothers who had drunk during pregnancy and their 20 children (ALD group), 17 alcoholic mothers who had not drunk during pregnancy and their 40 children (ALND group) and 60 non-alcoholic control mothers and their 80 children (Control group), were compared concerning the mothers' drinking problems and abnormal deliveries, children's birth weights, congenital abnormalities, abnormalities of the central nervous system and psychological problems. RESULTS: The mean age of onset of problem drinking of the mothers in the ALD group was significantly lower than that in the mothers of the ALND group, and some of the mothers in the ALD group showed alcohol dependence before their pregnancies. The mean birth weights of the children of the ALD group, ALND group and Control group were 2816 g, 3128 g and 3142 g, respectively and the differences were significant. The children of the ALD group had significantly more abnormal birth episodes, developmental retardation and psychiatric symptoms than those in the other two groups. Among 20 children in the ALD group, FASD was suspected in 6 children (10% of the children of alcoholic mothers). Six children had low birth weights, abnormal birth episodes, mental retardation and psychiatric symptoms. CONCLUSION: One third of the Japanese children of alcoholic mothers had experiences of prenatal alcohol exposure and 10% of them had suspected FASD abnormalities. PMID- 16038425 TI - [Actual condition of alcohol intake and its effects on lifestyle-related disease in health checkup]. AB - Actual condition of alcohol intake was investigated in forty four thousand one hundred and twenty six individuals who visited the Tokai University Hospital Health Checkup Center from 1989 to 2003. Effects of alcohol intake were also examined in relation to several risk factors for lifestyle-related diseases. The male drinkers who took more than 1 gou of sake per a day were recognized in 53.0% from 1989 to 1991, and decreased to 46.3% from 2001 to 2003. The female drinkers were found in approximately 10%, and remained unchanged during the 15-year survey period. When examined by age, the frequency of habitual drinking among males was 34.4% in the age of 20 years, and then increased to 45% in the 30 years, leading to the peak (54.1%) in the 40 years. In females, the frequency was 27.5% in the age of 20 years, but decreased to 10.9% in the 30 years. The prevalence of systolic hypertension, diastolic hypertension, hyperuricemia, high levels of HbAlc, and hypertriglyceridemia was significantly (P < 0.0001) increased with an increase in alcohol intake. The prevalence of obesity, fatty liver and hyperglycemia at fasting was markedly (P < 0.0001) increased in the drinkers whose intake was more than 2 gou per a day. These findings indicate that habitual drinking may be associated with risk factors for lifestyle-related diseases, such as obesity, fatty liver, hypertension, hypertriglyceridemia and hyperuricemia. PMID- 16038426 TI - [Assessment of microcirculation disturbance in patients with coronary ectasia by ATP-loading 99mTc-tetrofosmin myocardial SPECT]. AB - Patients with coronary ectasia often develop chest pain and reveal ischemic changes on electrocardiograms and reduced left ventricular wall motion on left ventriculography, in the absence of epicardial coronary artery stenotic regions. We examined the disturbances in the coronary microcirculation in patients with coronary ectasia using left ventriculography and ATP loading 99mTc-tetrofosmin myocardial single photon emission computed tomography (SPECT) before and after administration of a coronary vasodilator and antiplatelet agents. METHODS: Twenty patients in whom coronary angiography revealed diffuse coronary artery ectasia but no stenotic regions were enrolled in this study. Left ventriculography and ATP loading 99mTc-tetrofosmin myocardial SPECT were performed before and after administration of the coronary vasodilator, nicorandil, as well as that of the antiplatelet agents, aspirin and ticlopidine. RESULTS: (1) The ejection fraction in left ventriculography was 48.3 +/- 17.4% before, and 56.6 +/- 18.3% after the drug administration, the ejection fraction was improved after the drug administration (p < 0.05). (2) Before the drug administration, the total defect scores on 99mTc-tetrofosmin myocardial SPECT were 5.9 +/- 3.1 and 8.8 +/- 2.7 in the ATP-loading and rest images, respectively (p < 0.05), and the corresponding scores after the drug administration were 4.1 +/- 3.0 and 5.4 +/- 3.1, respectively (N.S.). Thus, the total defect scores in the ATP-loading and rest images improved after the drug administration (p < 0.05). CONCLUSION: Myocardial damage in patients with coronary ectasia might be induced by microthrombotic embolism and microcirculation disturbance. PMID- 16038427 TI - [Cost-utility analysis of antithyroid drug therapy versus 131I therapy for Graves' disease]. AB - There is no comparative cost-utility study between 131I therapy and antithyroid drugs (ATD) therapy for Graves' disease, though 131I therapy has higher remission rate and less side effects. The objective of the study was to analyze the cost utility of ATD therapy versus 131I therapy by calculating life-long medical costs and utility, based on the responses of Graves' disease patients to questionnaires. To determine the expected cost and expected utility, a decision tree analysis was designed on the basis of the 2 competing strategies of ATD therapy versus 131I therapy. A simulation of 1000 female patients weighing > or =50 kg who assumed to experience the onset of Graves' disease at the age of 30, to first complain of thyrotoxic symptoms and moderate goiter 2-3 mo. previously, and to undergo a 40-years-long cohort study, was created for each strategy using a decision tree and baselines of other relevant variables. The variables and costs were based on the literature and hospital bills. The maximum and minimum values of utility were defined as 1.0 and 0.0, respectively. Future costs and utilities were discounted 5%. The medical costs and utilities were 85,739-88,650 yen/patient/40 years and 16.47-16.56/patient/40 years, respectively, for the ATD therapy strategy, and 81,842 yen/patient/40 years and 17.41/patient/40 years, respectively, for the 131I therapy strategy. These results quantitatively demonstrated that the 131I therapy strategy was superior to the ATD therapy strategy in terms of both cost and utility. 131I therapy should be used more widely in Japan because of its greater utility and lower cost. PMID- 16038428 TI - [Feasibility of long-term outcome prediction in acute myocardial infarction using the discordance between early and delayed image on 123I-BMIPP myocardial scintigraphy]. AB - OBJECTIVES: The feasibility of long-term outcome prediction using BMIPP myocardial scintigraphy was evaluated in cases of acute myocardial infarction. METHODS: BMIPP myocardial scintigraphy was performed on 165 patients with first acute myocardial infarction at the time of discharge from the hospital (average of 27 days after disease on set). Discordance between early and delayed image was checked and its relation to later cardiac events (during the mean follow up period of 64.2 +/- 9.8 months) was analyzed. In 82 of these 165 cases TlCl scintigraphy was simultaneously performed (Tl/BMIPP dual SPECT) to examine mismatch form BMIPP scintigraphy and discordance between early and images. RESULTS: Discordance between early and delayed images was observed in 86 cases (52%). Among patients for whom dual SPECT was performed, mismatch between TlCl and BMIPP scintigraphy was observed in 30 cases (37%). When the relation between mismatch and discordance was analyzed, mismatch was accompanied by washout. The incidence of later cardiac events was significantly higher for cases showing discordance accompanied by washout and cases showing mismatch on dual SPECT scintigraphy than cases without these findings. When multivariate analysis was conducted, involving age, sex, infarction related artery, left ventricular end diastolic volume index, left ventricular ejection fraction, severity of disturbed fatty acid metabolism, washout and fill-in, washout was identified as an independent predictor of cardiac events. CONCLUSION: Mismatch on Tl/BMIPP dual SPECT is important for predicting long-term prognosis of acute myocardial infarction. Furthermore, washout on BMIPP scintigraphy is also useful as a predictor of cardiac events. PMID- 16038429 TI - [Analysis of brain images in patients with spinocerebellar degeneration; using statistical parametric mapping (SPM) and easy Z score imaging system (eZIS)]. AB - In order to investigate the cerebral blood flow objectively, the easy Z score imaging system (eZIS), was developed, and has been applied in clinical practice. SPECT with 99mTc-ethyl cysteinate dimer (99mTc-ECD) was performed, and the images were analyzed using the SPM97 and the eZIS Ver. 2 to investigate cerebral blood flow in patients with two types of spino-cerebellar degeneration. We compared the distribution of cerebral blood flow between 13 patients with cortical cerebellar atrophy (CCA) and 26 patients with olivopontocerebellar atrophy (OPCA). In the both groups, cerebellar blood flow was decreased generally. In our evaluation using the eZIS Z score, the scores for the brain stem and cerebellar nucleus in the OPCA group were lower than those in the CCA group. This method facilitates the objective evaluation of cerebral blood flow in patients with spinocerebellar degeneration, and may be useful for analyzing the condition of these disease. PMID- 16038430 TI - [Radioiodine treatment in patients with Graves' disease at outpatient clinic: special reference to safety and short-term outcome]. AB - PURPOSE: This retrospective study was aimed at revealing the safety and short term outcome of radioiodine treatment in patients with Graves' disease at outpatient clinic. METHODS: From July 1999 to April 2002, 511 patients with Graves' disease were treated with radioiodine at the outpatient clinic of Tajiri Thyroid Clinic, Kumamoto. Of them, 73 patients dropped out or were referred to another medical institution. In the remaining 438 patients [100 men and 338 women; 44.6 +/- 15.4 (mean +/- SD) (14-82) years old], the safety of radioiodine treatment at the outpatient clinic and the treatment outcome until April 2003 was examined. The dosage was determined based on radioactive iodine uptake (3 hours) and thyroid volume measured by ultrasound. The initial dosage was 6.7 +/- 3.3 (1.2-13.5) mCi. Five months later, it was evaluated whether or not radioiodine should be administered a second time. All patients were treated at the outpatient clinic. RESULTS: There was no particular problem associated with treatment. Patients with a large goiter could be successfully treated with divided doses. After 12-45 (30.1 +/- 9.3) months of radioiodine, thyroid function status was as follows; hyperthyroidism: 7 patients (1.6%), subclinical hyperthyroidism: 78 patients (17.8%), euthyroidism: 108 patients (24.7%), subclinical hypothyroidism: 116 patients (26.5%), hypothyroidism: 129 patients (29.4%). CONCLUSION: It was concluded that radioiodine treatment in patients with Graves' disease at outpatient clinic was safe and showed a satisfactory short-term outcome. PMID- 16038431 TI - [New therapies for inflammatory bowel disease]. PMID- 16038432 TI - [Diagnosis and treatment of bile duct cancer--from the viewpoint of physician]. PMID- 16038433 TI - [Surgical treatment and related diagnosis for bile duct cancer]. PMID- 16038434 TI - [Usefulness of branched-chain amino acid (BCAA)-enriched nutrient mixture for nutritional treatment undergoing endoscopic treatment for esophageal varices]. AB - We investigated the alteration of nutritional status in 144 patients who were treated for the first time with endoscopic sclerotherapy or endoscopic variceal ligation during their therapies. The serum levels of albumin, cholinesterase and total cholesterol were compared before and after treatment. The serum level of cholinesterase declined significantly. To investigate the impact of aging on the changes of nutritional status we divided all patients into two groups: (1) under 65 years, and (2) over 65 years. The decline of serum albumin of elderly patients (n=65) was significantly greater than that of younger patients (n=79). A branched chain amino acid (BCAA)-enriched nutrient mixture for nutritional treatment significantly suppressed the decline of serum albumin in elderly patients. Nutritional treatment with a BCAA-enriched nutrient mixture should be considered during endoscopic therapy for esophageal varices, especially in elderly patients. PMID- 16038435 TI - [A case of granulocyte-colony stimulating factor producing small cell carcinoma of esophagus]. PMID- 16038436 TI - [A case of simple ulcer with ulcerative colitis]. PMID- 16038437 TI - [A case of pneumatosis cystoides intestinalis developed in a patient with Sjogren syndrome during administration of anetholetrithion]. PMID- 16038438 TI - [A case of goblet cell carcinoid of the appendix presented as a metastatic ovarian tumor and peritoneal dissemination, and diagnosed by autopsy]. PMID- 16038439 TI - [Three cases of colorectal cancer in patients with Crohn's disease]. PMID- 16038441 TI - [A case of pediatric Crohn's disease with growth retardation]. PMID- 16038440 TI - [A case of malignant endocrine tumor of the pancreas showing atypical findings in diagnostic imaging]. PMID- 16038442 TI - [A case of peritoneal malignant mesothelioma that FDG-PET was useful for a supporting diagnosis]. PMID- 16038443 TI - [Evaluation of a new (13)CO2 infrared analyzer (POCone) for 13C-urea breath test]. PMID- 16038444 TI - [Researches of clinical anatomy performed by experienced surgeons and physicians coming from outside of the university]. AB - These 5 years, twenty experienced surgeons and physicians have come from outside of the university and performed researches of clinical anatomy in my laboratory. This report shows the specific research preparation, actual research life and related problems. I hope it will provide a new, fruiteful collaboration between the present author and other members of JAA. PMID- 16038445 TI - [Anatomy education in medical and dental schools in Japan]. AB - We studied the anatomy education and the view of anatomy professors on it in medical and dental schools in Japan. In most schools anatomy is taught in the second year. In medical schools, the systematic education separating macroscopic and microscopic anatomy is prevalent. Although the tutorial system has been introduced in 80% of medical schools, its introduction into anatomy education has remained at 30%. The tutorial system is regarded to be more effective by engaged professors than non-engaged. Some kinds of clinical anatomy education have been introduced in half of the medical schools surveyed. In dental schools, on the other hand, macroscopic and microscopic anatomy tend to be taught in combination. One third of the dental schools have introduced clinical anatomy but few schools have a tutorial system. The overwhelming majority of professors are evaluated by students and have regarded the evaluation useful for improving their teaching. They also have thought that the questionnaire and the timing of the evaluation must be considered carefully, and that the evaluation should not be directly used for purposes other than the improvement of education. We have made the proposals for further improvement in anatomy education based upon this study. PMID- 16038446 TI - The influence of automatic enrollment, catch-up, and IRA contributions on 401(k) accumulations at retirement. PMID- 16038447 TI - Language-specificity in the perception of paralinguistic intonational meaning. AB - This study examines the perception of paralinguistic intonational meanings deriving from Ohala's Frequency Code (Experiment 1) and Gussenhoven's Effort Code (Experiment 2) in British English and Dutch. Native speakers of British English and Dutch listened to a number of stimuli in their native language and judged each stimulus on four semantic scales deriving from these two codes: SELF CONFIDENT versus NOT SELF-CONFIDENT, FRIENDLY versus NOT FRIENDLY (Frequency Code); SURPRISED versus NOT SURPRISED, and EMPHATIC versus NOT EMPHATIC (Effort Code). The stimuli, which were lexically equivalent across the two languages, differed in pitch contour, pitch register and pitch span in Experiment 1, and in pitch register, peak height, peak alignment and end pitch in Experiment 2. Contrary to the traditional view that the paralinguistic usage of intonation is similar across languages, it was found that British English and Dutch listeners differed considerably in the perception of "confident," "friendly," "emphatic," and "surprised." The present findings support a theory of paralinguistic meaning based on the universality of biological codes, which however acknowledges a language-specific component in the implementation of these codes. PMID- 16038448 TI - Perception of place and secondary articulation contrasts in different syllable positions: language-particular and language-independent asymmetries. AB - This study investigated the perception of place and secondary articulation contrasts in different syllable positions by Russian and Japanese listeners. The consonants involved in the study were the Russian plain (velarized) and palatalized labial and coronal voiceless stops in syllable-initial and syllable final positions at word boundaries. The findings revealed substantial asymmetries in the perception of the contrasts by both groups of listeners: With respect to positions, consonants in syllable-final position were characterized by lower correct identification rates and (less consistently) longer reaction time than the same consonants in syllable-initial position. Positional syllable position differences were accompanied by differences in segment-specific contexts. With respect to individual consonants, the palatalized labial /p(j)/ and the plain coronal /t/ showed a lower correct identification rate and smaller perceptual distance than the plain labial /p/ and the palatalized coronal /t(j)/. The results also showed some differences between Russian and Japanese listeners in the perception of the contrast. These differences can be explained by phonotactic differences between the two languages, as well as by differences in the phonetic realizations of the consonants. The results of the study provide evidence for the role of both universal and language-particular factors in speech perception. PMID- 16038449 TI - Automatic discrimination of emotion from spoken Finnish. AB - In this paper, experiments on the automatic discrimination of basic emotions from spoken Finnish are described. For the purpose of the study, a large emotional speech corpus of Finnish was collected; 14 professional actors acted as speakers, and simulated four primary emotions when reading out a semantically neutral text. More than 40 prosodic features were derived and automatically computed from the speech samples. Two application scenarios were tested: the first scenario was speaker-independent for a small domain of speakers while the second scenario was completely speaker-independent. Human listening experiments were conducted to assess the perceptual adequacy of the emotional speech samples. Statistical classification experiments indicated that, with the optimal combination of prosodic feature vectors, automatic emotion discrimination performance close to human emotion recognition ability was achievable. PMID- 16038450 TI - [Expression of nestin and glial fibrillary acidic protein in injured spinal cord of adult rats at different time]. AB - OBJECTIVE: To explore the expression of nestin and glial fibrillary acidic protein (GFAP) at different time and sites after spinal cord injury in adult rats. METHODS: Seventy-two adult Sprague-Dawley rats, aging 8 weeks and weighing from 180 to 220 g, were randomly divided into 11 experimental groups (66, n=6) and 1 control group (n= 6). In the experimental groups, the rat spinal cord injury models were established by aneurysm clip compression, and the expression and proliferation of nestin and GFAP at different time (1 day, 3 days, 5 days, 1 week, 2 weeks, 3 weeks, weeks, 5 weeks, 6 weeks, 7 weeks and 8 weeks) and at different sites (injured site and adjacent site) were observed with toluidine blue staining, immunofluorescent staining and the analytical system of photographs. In control group, the same site of the rat spinal cord was exposed without aneurysm clip compression. The same preparation and examination were done as the experimental groups. RESULTS: Toluidine blue staining results showed that contour of neurite and pericaryon were distinct and nucleus were deep blue in normal control rats. One day after injury, the number of big and medium-sized neuron decreased obviously; neurite was deep blue with clouding Nissl bodies and ellipse or triangular typed nucleus. In the normal control group, the expression of nestin was hardly seen except ependymal cells of central canal, and the low expression of GFAP was seen. In the experimental groups, the nestin and GFAP expressions increased obviously in the injured sites and adjacent sites 24 hours after injury, reached the peak value after 3-7 days and followed by gradual decrease. There were statistically significant differences in the nestin and GFAP expressions between the experimental groups and the control group. CONCLUSION: The above results suggest that spinal cord injury can induce the expression of nestin and GFAP. There is a positive correlation between nestin expression and the proliferation of the reactive astrocytes. PMID- 16038451 TI - [Effects of neural stem cells transplantation on glial cell line-derived neurotrophic factor and growth associated protein 43 after spinal cord injury in rats]. AB - OBJECTIVE: To observe the effects of neural stem cells(NSCs) transplantation on the glial cell line-derived neurotrophic factor (GDNF) and growth associated protein 43(GAP-43) after the spinal cord injury (SCI), and to investigate the mechanism of repairing the SCI by NSCs transplantation. METHODS: The neural stem cells from the hippocampus of rats' embryo were cultured and identified by immunocytochemistry. The SCI model was made by the modified Allen device. Sixty adult Wistar rats were randomly divided into three groups: spinal cord injury was treated with transplantation of NSCs (group A, n = 24), with DMEM solution (group B, n = 24) and normal control group without being injured(group C, n = 12). Seven days after the operation of SCI, the NSCs were transplanted into the injured site. Then GAP-43 and GDNF expressions were tested by RT-PCR and immunohistochemistry. RESULTS: Compared with group B, the GDNF mRNA expression of group A increased by 23.3% on the 1st day, by 26.8% on the 3rd day and by 32.7% on the 7th day; the GAP-43 mRNA expression increased by 19.5% on the 1st day, 21.6% on the 3rd day and 23.1% on the 7th day. There were statistically significant differences (P<0.05). CONCLUSION: The transplantation of NSCs can change the micro-environment injured site and promote the regeneration of axon by enhancing the expressions of GDNF mRNA and GAP-43 mRNA. It is one of the mechanisms of repairing the SCI by NSCs transplantation. PMID- 16038452 TI - [Application of freeze-dried cancellous allograft in treatment of spinal tuberculosis]. AB - OBJECTIVE: To investigate the efficacy of freeze-dried cancellous allograft in the treatment of spinal tuberculosis. METHODS: From January 1999 to August 2004, there were 31 cases of spinal tuberculosis who underwent surgery. The freeze dried cancellous allograft was used as grafting material in all the cases. The cancellous allograft was packed in a titanium mesh cage or an artificial vertebrae, and then used as a strut graft anteriorly to implant into the bone defect after the radical debridement, and the instrumentation was done. RESULTS: Twenty-three cases were followed up 1.5 years to 5 years (3.7 years on average), and bony fusion was achieved in 21 cases 6 months later. In 2 cases ceasing antituberculous therapy after 2 months of operation, the local recurrence was obvious. The loosened screw was noticed in one of these two cases, who had tuberculosis in lumbar spine. When antituberculous therapy continued, the bony fusion was observed in these two cases 12 months later. No further position change of the instrument was noticed in the patient carrying loosened screw, but the kyphosis of the thoracolumbar spine aggravated. CONCLUSION: Freeze-dried cancellous allograft could be used in the treatment of spinal tuberculosis. To achieve good results of allograft incorporation and remodeling, the rigid instrumentation should be performed, postoperative antituberculous therapy is also important. PMID- 16038453 TI - [Diagnosis and treatment of multiple-level noncontiguous spinal fractures]. AB - OBJECTIVE: To explore the injury mechanism, clinical features and treatment methods of multiple-level noncontiguous spinal fractures (MNSF). METHODS: The clinical data of 23 patients with MNSF were analyzed retrospectively. There were 15 males and 8 females aging from 13 to 75 years. Five cases missed diagnosis. The locations of the primary injury were cervical region in 3 cases, thoracic region in 7 cases, and thoracolumbar region in 13 cases. The spinal fractures associated with spinal cord injury were 14 cases in the primary injury, and 3 cases in the secondary injury. According to Frankel grade, there were 7 cases of grade A, 1 case of grade B, 3 cases of grade C, 6 cases of grade D and 6 cases of grade E. Sixteen cases were treated by surgical stabilization while 7 cases were treated conservatively. RESULTS: Twenty patients were followed up from 3 months to 4 years with a mean of 11.5 months except three patients. No neurologic deterioration was observed in patients who were treated conservatively or operatively. No wound infection, no implant failure or bone graft nonunion occurred in patients who were treated operatively. Among 17 patients with neurologic deficit before operation, 12 got 1-3 grade improvement of the Frankel grading, 4 remained unchanged and 1 died. CONCLUSION: The mechanism of MNSF injury is complex, it is easy to miss diagnosis. The patients with spinal injury must be examined carefully and completely. A whole spine radiographic survey should be accomplished if necessary and treatments should be based on the stability of spine fractures and the severity of spinal cord injury. PMID- 16038454 TI - [Stress changes of upper lumbar facet joint after discectomy and artificial disc replacement]. AB - OBJECTIVE: To explore the influence of discectomy and artificial disc replacement on the upper lumbar facet joint and to provide the evidence for use of artificial disc replacement. METHODS: Under the loads of 200-2000 N axis pressure and 1-10 Nm back-extending/side-bending pressure on the 7 fresh adult corpses, the pressure of L3.4 facet joint was measured in the case of L4.5 disc integrity, discectomy and artificial disc replacement and the statistical analysis was done. RESULTS: Under the same load (axis, back-extending/side bending), there were significant differences (P<0. 01) in the pressure of upper lumbar facet joint both between disc integrity and discectomy and between discectomy and artificial disc replacement, and there was no difference (P>0. 05) between disc integrity and artificial disc replacement. CONCLUSION: The stress of upper lumbar facet joint reduced after discectomy, it implies that simply discectomy may change biomechanics of lumbar and lead to secondary lumbar structure degeneration. The stress of upper lumbar facet joint is close to the normal value after artificial disc replacement, it implies that artificial disc replacement may protect lumbar structure. It provides some biomechanics foundation and evidence for artificial disc replacement. PMID- 16038455 TI - [Repair of bone defect with allograft demineralized bone containing basic fibroblast growth factor in rabbits]. AB - OBJECTIVE: To evaluate the ability of inductive osteogenesis of allograft demineralized bone containing basic fibroblast growth factor (bFGF/ALB) in repairing bone defect. METHODS: Thirty-two New Zealand white rabbits were randomly divided into four groups (groups A,B,C and D, n=8). A segmental bone defect of 15 mm in length was made on the bilateral radius respectively and the defects filled with ALB/bFGF in group A, with ALB in group B, with bFGF in group C and without any materials in group D serving as blank control. At 2, 4, 6 and 8 weeks after operation, all restored bones were evaluated by roentgenography, histological observation and Ca2+ detection of osteotylus. RESULTS: The X-ray films showed that groups A and B had a little shadow of bone formation at 2 weeks, while groups C and D had transparent shadow; that group A had denser shadow and new bone formation at 4 weeks and 6 weeks, groups B and C had a little increase of shadow and group D had little shadow at fractured ends; and that group A had formation of bone bridge at 8 weeks, the new formed bone in fractured ends of group B closed with each other, the gap still existed in group C, and the defects filled with the soft tissue in group D. The Ca2+ content of group A was higher than that of groups B, C and D at 4 weeks (P<0.05) and 8 weeks (P<0.01). The histological observation showed that the degree of bone restoration of group A was superior to that of groups B, C and D. CONCLUSION: bFGF/ALB is a good material to improve bone restoration. PMID- 16038456 TI - [Expression of vascular endothelial growth factor in repairing bone defect with vascularized bone graft- reconstituted bone xenograft]. AB - OBJECTIVE: To study efficiency of vascularized bone graft combining with reconstituted bone xenograft (RBX) in repairing bone defect and the expression of the vascular endothelial growth factor (VEGF) in serum. METHODS: From January 1998 to December 2002, 27 cases of bones defects were treated and randomly divided into 3 groups according to different repair materials: group A (the vascularized bone graft- RBX group, n=9), group B (the vascularized bone graft group, n=10) and group C(the RBX group, n=8). The bone defect repair, the bone healing time and the bone graft resorption were observed by radiograph after 3 months, 6 months and 12 months of operation, and the expression of VEGF in serum was assayed with lumino-enzyme immunoassay before operation and after operative 2 weeks, 4 weeks, 6 weeks and 8 weeks respectively. RESULTS: The X-ray films showed that the bone healing was achieved in 8 cases of group A, in 6 cases of group B and in 3 cases of group C after 3 months; in 1 case of group A, respectively in 3 cases of both group B and group C after 6 months. The bone graft resorption was observed in 1 case of group B and in 2 cases of group C after 12 months. The serum VEGF values after operative 2 weeks and 4 weeks were higher than those before operation in all of 3 groups (P<0. 05), and the VEGF values of groups A and B were higher than that group C(P<0. 05) after 4 weeks. There were no significant differences (P>0. 05) in serum VEGF level between postoperative 6, 8 weeks and pre-operation in 3 groups. CONCLUSION: The expression of serum VEGF obviously increase in the early period of bone transplanting, it is value of clinical evaluation of reparative efficiency of bone defect. PMID- 16038458 TI - [Treatment of ischemic necrosis of the femoral head by transplantation of pedicled bone flap with deep iliac circumflex vessel]. AB - OBJECTIVE: To investigate the treatment method of ischemic necrosis of the femoral head by the transplantation of pedicled bone flap with deep iliac circumflex vessel in adult and to assess its curative effect. METHODS: From February 1996 to September 2003, 46 adult patients with ischemic necrosis of the femoral head (58 coxas) were treated by transplantation of pedicled bone flap with deep iliac circumflex vessel. The locations were the left side in 34 coxas and the right side in 24 coxas(stage II in 16 coxas, stage III in 39 coxas and stage IV in 3 coxas). RESULTS: Thirty-seven patients (40 coxas) were followed up 6 months to 10 years after operation. According to the assess of FanQishen, the result was excellent in 19 cases (21 coxas), good in 11 cases (12 coxas), fair in 5 cases (5 coxas) and poor in 2 cases (2 coxas). The postoperative excellent and good rate was 81%. CONCLUSION: Operative treatment of ischemic necrosis of the femoral head in adult by transplantation of bone flap pedicled with deep iliac circumflex vessel has the characteristics of restoration of blood supply of the femoral head, decompression of marrow cavity, elimination of necrotic bone and support of the femoral head. It is an effective and advanced method for treatment of ischemic necrosis of the femoral head in adult. PMID- 16038457 TI - [An experimental study on repairing bone defect with the biodegradable polycaprolactone material]. AB - OBJECTIVE: To investigate the ability of the biodegradable polycaprolactone (PCL) material to repair bone defect and to evaluate the feasibility of using the PCL as the scaffold in tissue engineering bone. METHODS: The bone defect models of 4. 5 mm x 12 mm were made in the bilateral femoral condyle of 65 New Zealand white rabbits. The PCL cylinder was implanted into the right side of defect (experimental group, n = 60), the high dense hydroxyapatite was implanted into the left side of defect (control group, n = 60), and the incision was sutured without any implants (blank group, n = 5). The samples were harvested and observed by examinations of gross, X-ray, bone density, 99mTc-MDP bone scanning, gamma-display ratio and scanning electron microscope (SEM) after 3, 6, 9 and 12 months of operation. The results were compared between the experimental group and control group. RESULTS: At 3, 6, 9 and 12 months after operation, the gross and X ray examinations indicated that the bone defect filled with the new bone on the PCL-tissue surface, and no delayed inflammatory reaction appeared. The average bone mineral density was greater in the experimental group than that in the control group, and the difference had statistical significance (P< 0. 05). The results of 99mTc-MDP bone scanning and gamma-display ratio showed that the nuclide uptake was more in the PCL group than that in the control group. The SEM result proved that the new compact bone formed on the PCL migrating surface as the PCL degraded gradually,but the collagen fiber sheath formed around the hydroxyapatite in the control group. CONCLUSION: PCL possesses good biocompatibility and high bone inductive potentiality, it can be used to repair bone defect. PMID- 16038459 TI - [A comparative study on operation and non-operation in treating fractures of scapular neck]. AB - OBJECTIVE: To explore the effective method for treatment of fractures of scapular neck. METHODS: A retrospective analysis of 30 patients with fractures of scapular neck was done. The patients were treated by operation or by non-operation from May 1994 to May 2003. According to the degree of displacement, 30 cases included 9 mild displaced fractures(<10 mm) and 21 severe displaced fractures(>10 mm). According to the location of fractures, there were 24 cases of surgical neck fractures and 6 cases of anatomical neck fractures. Of all 30 patients, 12 were treated by non-operation and 18 by operation. All fractured scapular necks were exposed through posterior approach and fixed with interfragmentary compression screws, reconstruction plates and so on after satisfactory reduction. Associated clavicle fractures were exposed through Langer's approach and fixed with Kirschner wire-tension band fixation construct, reconstruction plates and so forth. When followed up, the clinical examination was done and the X-ray films were taken to measure glenopolar angle(GPA). Herscovici's score was adopted. RESULTS: Twenty-eight patients were followed up from 14 days to 9 years. Among 11 patients treated by non-operation, Herscovici score showed that 4 cases achieved the excellent result, 2 cases good, 2 cases fair and 3 cases poor and the X-ray film results showed that there were 6 cases of GPA>20 degrees and 5 cases of GPA<20 degrees. Of 17 patients treated by operation, Herscovici score showed that 11 cases achieved the excellent result, 5 cases good and 1 case fair and the X ray film results showed that there 16 cases of GPA>20 degrees and 1 case of GPA<20 degrees . CONCLUSION: Fractures of scapular surgical neck with mild displacement can be treated by non-operation, while fractures of surgical neck with severe displacement or associated with ipsilateral clavicle fractures, and fractures of anatomical neck should be treated by operation to reconstruct the stability of shoulder and minimize the complications. PMID- 16038460 TI - [Clinical application and efficiency of two stage multiple nerves transfer for treatment of root avulsion of brachial plexus]. AB - OBJECTIVE: To investigate the results of two stage multiple nerves transfer for treatment of complete brachial plexus root avulsion. METHODS: Eight patients with complete brachial plexus avulsion, aging 18-38 years and with a mean 6 months interval of injury and repair, were surgically treated with the following procedures. One stage surgical procedure was that the contralateral C7 never root was transferred to the ulnar nerve, the phrenic nerve to the anterior division of upper trunci plexus brachialis and the accessory nerve to the suprascapular nerve. Two stage surgical procedure was that the ulnar nerve was transferred to the median nerve , the intercostal nerves to the radial nerve and the thoracodorsal nerve. RESULTS: All patients were followed up from 13 months to 25 months (21 months on average), muscle reinnervation was observed in all patients. Return of muscle power of M3 or better are regarded as effective. The effective recovery results were 75% in musculocutaneous nerve, 37.5% in suprascapular nerve, 37.5% in radial nerve, 75% in thoracodorsal nerve and 62.5% in median nerve. In sensory recovery of the median nerve, 4 patients obtained S3, 3 patients S2 and 1 patient S1. CONCLUSION: Two stage multiple nerves transfer for treatment of root avulsion of brachial plexus can achieve better motor function results and is safe and effective. The procedure should be recommended for treatment of root avulsion of brachial plexus in selected patients with complete brachial plexus root avulsion, especially in young patients with a short interval between injury and repair. It is one of the alternative options. PMID- 16038461 TI - [Repair of defect on finger skin with reverse fascial pedicle island flap of dorsal branch of digital artery]. AB - OBJECTIVE: To introduce the surgical procedure and indication of the reverse fascial pedicle island flap of the digital artery dorsal branches in repairing finger skin defect. METHODS: By use of the dorsal branches of the digital artery as the pedicel, the reverse island flap was designed. The skin defects of the proximal interphalangeal joint and beyond in 35 cases (42 fingers) were repaired and the joint or extensor tendon was reconstructed simultaneously. Donor site was primarily closed or a skin graft was used. The flap size ranged from 1.0 cm x 2.5 cm to 1.5 cm x 3.5 cm. RESULTS: Thirty-five patients were followed up 3 months to 1 year, all the flaps survived. The two-point discrimination was between 6 mm and 10 mm. The function of interphalangeal joint was satisfactory. CONCLUSION: These flaps have the advantages of an extended skin paddle and a versatile pivot point on the phalanx, and they allow coverage of wide and distal defects. PMID- 16038462 TI - [Repair of anterior tibial, dorsal pedal and calcaneal soft-tissue defects with lateral crural flaps]. AB - OBJECTIVE: To report the methods and clinical effect of the lateral crural flaps in repairing anterior tibial, dorsal and calcaneal soft-tissue defects. METHODS: From August 1999 to December 2004, 18 cases of defects were repaired with lateral crural flap, including 15 cases of anterior tibial, dorsal and calcaneal soft tissue defects with vascular pedicled island lateral crural flaps and 3 cases of dorsal pedal soft-tissue defects with free vascular lateral crural flaps. RESULTS: All flaps survived after operation. Insufficient arterial supply of the flap occurred in 2 cases after operation, the pedicled incision sewing thread was removed and lidocaine was injected around vascular pedicle, then the flap ischemia was released. Inadequate venous return and venous hyperemia occurred in 1 case because peroneal vein was injured during operation. The flap edge skin was cut and heparin was locally dripped for one week, the flap vascular cycle was resumed. All patients were followed up two months to one year, the flaps were not fat, and the elasticity was good. CONCLUSION: It is safe and reliable to use lateral crural flap to repair anterior tibial, dorsal pedal and calcaneal soft tissue defects. PMID- 16038463 TI - [Repair of distal urethral stricture after urethroplasty of hypospadias]. AB - OBJECTIVE: To search for a new method to repair distal urethral stricture resulting from urethroplasty of hypospadias. METHODS: From February 2000 to March 2004, 16 patients with distal urethral stricture were treated by use of cutting stricture urethra and their distal urethra were reconstructed with phallic flap. RESULTS: All operations were successful without complication of flap necrosis. After 7 days of operation, the patients had free micturition and thick stream of urine. Eleven patients were followed 2 months to 4 years, the satisfactory result was obtained. CONCLUSION: It is a simple and good method to reconstruct the distal urethra by superimposing the phallic flap on the cut stricture urethra after urethroplasty of hypospadias. PMID- 16038464 TI - [Morphological and biomechanical study on in vivo osteogenesis after repair of cranial defects with plastic engineered bone in rabbits]. AB - OBJECTIVE: To investigate the morphology and biomechanics of in vivo osteogenesis after repairing rabbit skull defects with plastic engineered bone which was prefabricated with alginate gel, osteoblasts and bone granules. METHODS: Twenty eight rabbits were divided into group A (n=16), group B (n=8) and group C (n=4). The bilateral skull defects of 1 cm in diameter were made. Left skull defects filled with alginate gel-osteoblasts-bone granules (group A1) and right skull defects filled with alginate gel-bone granules (group A2). The defects of group B was left, as blank control and group C had no defect as normal control. The morphological change and bone formation were observed by methods of gross, histology and biomechanics. RESULTS: In group A1, the skull defects were almost entirely repaired by hard tissue 12 weeks after operation. The alginate gel osteoblasts-bone granule material had changed into bone tissue with few bone granules and some residuary alginate gel. The percentage of bone formation area was 40.92% +/- 19.36%. The maximum compression loading on repairing tissue of defects was 37.33 +/- 2.95 N/mm; the maximum strain was 1.05 +/- 0.20 mm; and loading/strain ratio was 35.82 +/- 6.48 N/mm. In group A2, the alginate and bone granules material partially changed into bone tissue 12 weeks after operation. The percentage of bone formation area was 18.51% +/- 6.01%. The maximum compression loading was 30.59 +/- 4.65 N; the maximum strain was 1.35 +/- 0.44 mm; and the loading/strain ratio was 24.95 +/- 12.40 N/mm. In group B, the skull defects were mainly repaired by membrane-like soft tissue with only few bone in marginal area; the percentage of bone formation area was 12.72% +/- 9.46%. The maximum compression loading was 29.5 +/- 2.05 N; the maximum strain was 1.57 +/- 0.31 mm; and the loading/strain ratio was 19.90 +/- 5.47 N/mm. In group C, the maximum compression loading was 41.55 +/- 2.52 N; the maximum strain was 0.95 +/- 0.17 mm; and the loading/strain ratio was 47.57 +/- 11.22 N/mm. CONCLUSION: The plastic engineered bone prefabricated with alginate gel-osteoblasts-bone granule may shape according to the bone defects and has good ability to form bone tissue, whose maximum compression loading can reach 89% of normal skull and the hardness at 12 weeks after operation is similar to that of normal skull. PMID- 16038465 TI - [Experimental studies on preparation and property of scaffold material of bio derived bone loading wo-1]. AB - OBJECTIVE: To provide the chosen scaffold materials for experiment and application of tissue engineering and to detect the properties of the collagen bio-derived bone scaffold material loading WO-1. METHODS: The pure bio-derived bone scaffold material, bio-derived bone scaffold material loading collagen, collagen bio-derived bone scaffold material loading WO-1 were made by use of allograft bone, and type-I collagen, and WO-1. The morphological features, constitute components and mechanical properties were examined by scanning electron microscopy, X- rays diffraction and mechanical assay. RESULTS: The bio derived bone scaffold material maintained natural network pore system; the bio derived bone scaffold material loading collagen maintained natural network pore system, the surface of network pore system was coated by collagen membrane; the collagen bio-derived bone scaffold material loading WO-1 maintained natural network pore system, the surface of network pore system was coated by collagen membrane. The pore sizes of the 3 materials were 90-700 microm, 75-600 microm and 80-600 microm, respectively, and the porosities were 87.96%, 80.47%, 84.2%. There was no significant difference between them (P>0.05). The collagen bio-derived bone scaffold material loading WO-1 consisted of [HA,Ca10 (OH)2 (PO4)6. There was no significant difference in the mechanical strength of the three scaffold materials. CONCLUSION: The bio-derived bone scaffold material loading WO-1 is as good as bio-derived bone scaffold material and collagen bio-derived bone scaffold material, and it is an effective scaffold material for tissue engineering bone. PMID- 16038466 TI - [An experimental study on repairing bone defect with composite of beta-tricalcium phosphate-hyaluronic acid-type I collagen-marrow stromal cells]. AB - OBJECTIVE: To observe the ability to repair bilateral radius bone defect with the composite of beta-tricalcium phosphate (PTCP), hyaluronic acid (HA), type I collagen (COL-I) and induced marrow stromal cells (MSCs), and to investigate the feasibility of the composite as a bone substitute material. METHODS: The MSCs of the New Zealand white rabbits were induced into osteoblasts, then combined with beta-TCP, HA and COL-I. Thirty New Zealand white rabbits were made the bilateral radius bone defects of 2 cm and divided into groups A, B and C. After 8 weeks, beta-TCP-HA-COL-I-MSCs (group A, n=27 sides), autograft (group B, n=27 sides)and no implant (group C as control, n=6 sides) were implanted into the areas of bilateral radius bone defects, respectively. The structure of the composite was observed by scanning electron microscope. The repairing effect was observed by gross, histomorphology, X-ray examination, and the degradation rate of inorganic substance at 4, 8 and 12 weeks. The osteogenic area and biomechanics of group A were compared with those of group B at 12 weeks. RESULTS: The MSCs could stably grow in vitro, relatively rapidly proliferated, and could be induced into the osteoblasts. The composite was porous. The results of gross, histomorphology and X-ray showed that the bone defects were perfectly repaired in group A and group B, but not in group C. The osteogenic area or biomechanics had no statistically significant difference between groups A and B (P> 0.05). The weight of inorganic substance in group A were 75%, 57% and 42% at 4, 8, 12 weeks, respectively. CONCLUSION: MSCs can be used as seed cells in the bone tissue engineering. The composite has porous structure, no reactions of toxicity to the tissue and rapid degradation, and it is an ideal carrier of seed cells. The beta-TCP-HA-COL-I-MSCs composite has the high ability of repairing bone defect and can serve as an autograft substitute material. PMID- 16038467 TI - [Experimental study on gelatin-chondroitin sulfate-sodium hyaluronate tri copolymer as novel scaffolds for cartilage tissue engineering]. AB - OBJECTIVE: To fabricate a novel gelatin-chondroitin sulfate-sodium hyaluronate tri-copolymer scaffold and to confirm the feasibility of serving as a scaffold for cartilage tissue engineering. METHODS: Different scaffolds was prepared with gelatin-chondroitin sulfate-sodium hyaluronate tri-copolymer by varying the freezing temperatures (-20 degrees C, - 80 degrees C and liquid nitrogen). Pore size, porosity, inter pores and density were observed with light microscopy and scanning electron microscopy (SEM). The load-stiffness curves were compared between different scaffolds and normal cartilage. The number of MSCs attaching to different scaffolds and the function of cells were also detected with MTT colorimetric microassay. RESULTS: The pore size was 300 +/- 45, 230 +/- 30 and 45 +/- 10 microm; the porosity was 81%, 79% and 56%; the density was 9.41+/-0.25, 11.50 +/- 0. 36 and 29.50 +/- 0. 61 microg/mm3 respectively in different scaffolds fabricated at - 20 degrees C, - 80 degrees C and liquid nitrogen; the latter two scaffolds had nearly the same mechanical property with normal cartilage; the cell adhesion rates were 85.0%, 87.5% and 56.3% respectively in different scaffolds and the scaffolds can mildly promote the proliferation of MSCs. CONCLUSION: Gelatin-chondroitin sulfate-sodium hyaluronate tri-copolymer scaffold fabricated at -80 degrees C had proper pore size, porosity and mechanical property. It is a novel potential scaffold for cartilage tissue engineering. PMID- 16038468 TI - [Repair of thyroid cartilage defects with chondrocyte-allogenous acellular cartilaginous matrix composite in rabbits]. AB - OBJECTIVE: To investigate the feasibility of repairing thyroid cartilage defects by implantation of chondrocyte-allogenous acellular cartilaginous matrix (chondrocyte-ACM) composite in rabbits. METHODS: The thyroid chondrocytes were isolated and co-cultured in vitro with allogenous acellular cartilaginous matrix (ACM) to form the chondrocyte-ACM composite. The composite was analyzed histologically and was used to repair defects of thyroid cartilage. Eighteen New Zealand adult rabbits were made the defect models of thyroid cartilage at the two sides and divided into three groups. The defects were repaired with chondrocyte ACM composite in the experimental group (n= 6), with simple ACM in the ACM group (n=6) and without any material in the control group (n=6). The animals were sacrificed at 8 weeks after operation. The specimens were evaluated histologically. RESULTS: In vitro, the growth of chondrocytes was observed on the surface of allogenous acellular cartilaginous matrix and no chondrocytes grew inside the matrix. The defect filled with muscle and connective tissues in control group; the lymphocyte infiltration was observed in the matrix and no new cartilage formation occurred at 8 weeks after operation in simple ACM group and experimental group. So the defect repair of rabbits thyroid cartilage failed. CONCLUSION: The allogenous acellular cartilaginous matrix failed to serve as a scaffold for chondrocytes both in vitro and in vivo. The allogenous acellular cartilaginous matrix should be improved. PMID- 16038469 TI - [Effect of surface property of different polyether-ester copolymers on growth of smooth muscle cells and endothelial cells]. AB - OBJECTIVE: To investigate the effect of surface property of different polyether ester block copolymers [poly(ethylene glycol-terephthalate)/poly(butylene terephthalate), PEGT/PBT] on the growth of smooth muscle cells (SMCs) and endothelial cells(ECs). METHODS: Three kinds of copolymers were synthesized, which were 1000-T20 (group A), 1000PEGT70/PBT30 (group B) and 600PEGT70/PBT30 (group C). The water-uptake and contact angle of three polyether-ester membranes were determined. The canine aorta smooth muscle cells and external jugular vein endothelial cells were primarily harvested, subcultured, and then identified. The proliferation of SMCs and ECs on the different polyether-ester membranes were investigated. RESULTS: The water-uptake of three copolymers arranged as the sequence of group Cgroup A> group B, indicating group B being more hydrophilic. However, smooth muscle cells and endothelial cells grew poorly on the membrane of group B after low density seeding, but proliferated well on the membranes of group A and group C. CONCLUSION: In contrast with more hydrophilic 1000PEGT70/PBT30, moderately hydrophilic 1000-T20 and 600PEGT70/PBT30 has better compatibility with vascular cells. The above results indicate that the vascular cells can grow well on moderately hydrophilic PEGT/PBT and that PEGT/PBT can be used in vascular tissue engineering. PMID- 16038470 TI - [Study on human leukocyte antigen G1 reducing xeno-cell-rejection by transfecting porcine endothelial cells]. AB - OBJECTIVE: To study whether the porcine endothelial cells (PECs) lines transfected by HLA-G1 can alter the lysis mediated by human peripheral blood mononuclear cell (PBMC) and natural killer cell 92 (NK-92). METHODS: By use of liposomes pack, the pcDNA3. 0 eukaryotic expression vector carrying HLA-G1 was transfected into PECs. Using indirect immunofluorescence and RT-PCR assays, the HLA-G1 expression in PECs was detected. The alteration of the lysis mediated by PBMC and NK-92 was detected by 51Cr-release assays. RESULTS: HLA-G1 expression could be detected in PECs after transfection of HLA-G1 at the levels of protein and RNA. It also could be found that the survival rate of transfected PECs was much higher than that of non-transfected PECs, when both of them faced the lysis mediated by human PBMC and NK-92. After transfecting the expression of HLA-G1 could be found in the transfected PECs and the lysis mediated by PBMC and NK-92 to PECs decreased obviously (P<0.05). CONCLUSION: The PECs transfected by HLA-G1 can decrease the NK lysis, so that it may provide us a new thought to inhibit the xeno-cell-rejection. PMID- 16038471 TI - [Roles of glial cell line-derived neurotropic factor in motoneuron development and motoneuron disease]. AB - OBJECTIVE: To review research progress of the relation between glial cell line derived neurotropic factor (GDNF) and motor neuron development and motor neuron disease. METHODS: The recent articles on GDNF and motor neurons were extensively reviewed. The molecular structure, the mode of action and the route of administration of GDNF were investigated. RESULTS: GDNF plays extensive roles in the development and disease of motor neuron. GDNF might regulate the development of the motor neurons of the spinal cord to some extent and also save the injured motor neurons. CONCLUSION: GDNF has a potential clinical value and inestimable future in the treatment of motor neuron diseases. PMID- 16038472 TI - Therapeutic drug monitoring and drug-drug interactions involving antiretroviral drugs. AB - The consensus of current international guidelines for the treatment of HIV infection is that data on therapeutic drug monitoring (TDM) of non-nucleoside reverse transcriptase inhibitors (NNRTIs) and protease inhibitors (Pls) provide a framework for the implementation of TDM in certain defined scenarios in clinical practice. However, the utility of TDM is considered to be on an individual basis until more data are obtained from large clinical trials showing the benefit of TDM. In April 2004, a panel of experts met for the second time in Rome, Italy. This was following the inaugural meeting in Perugia, Italy, in October 2000, which resulted in the manuscript published in AIDS 2002, 16(Suppl 1):S5-S37. The objectives of this second meeting were to review and update the numerous questions surrounding TDM of antiretroviral drugs and discuss the clinical utility, current concerns and future prospects of drug concentration monitoring in the care of HIV-1-infected individuals. A major focus of the meeting was to discuss and critically analyse recent and precedent clinical drug-drug interaction data to provide a clear framework of the pharmacological basis of how one drug may impact the disposition of another. This report, which has been updated to include material published or presented at international conferences up to the end of December 2004, reviews recent pivotal pharmacokinetic interaction data and provides advice to clinical care providers on how some drug drug interactions may be prevented, avoided or managed, and, when data are available, on what dose adjustments and interventions should be performed. PMID- 16038473 TI - Comparison of tests and procedures to build clinically relevant genotypic scores: application to the Jaguar study. AB - OBJECTIVE: To compare non-parametric tests and procedures of selection in building clinically validated genotypic scores. DESIGN AND PATIENTS: In the Jaguar study, 111 patients on a stable antiretroviral regimen experiencing virological failure were randomized in the didanosine (ddI) arm to receive ddI for 4 weeks in addition to their current combination therapy. METHODS: The virological response was HIV-1 RNA reduction from baseline to week 4. The univariate impact of each mutation associated with resistance to ddI on virological response was quantified by comparing reduction in plasma HIV-1 RNA in patients with or without the specific mutation, using a Wilcoxon-Mann-Whitney test. The next step was to select the combination of mutations most strongly associated with the virological response. Two procedures and two tests were compared using either the set of resistance mutations or the set of resistance mutations and mutations providing a better virological response. The Kruskal Wallis and the Jonckheere test for ordered alternatives were compared in order to build a genotypic score using the two distinct procedures. RESULTS: Eight mutations were associated with a reduced virological response to ddI: M41L, D67N, T69D, L74V, V1181, L210W, T215Y/F and K219Q/E and two mutations with a better virological response: K70R and M184V/I. The Jonckheere-Terpstra test for trend provided the combination of mutations (M41L+T69D-K70R+L74V-M184V /I+T215Y/F+ K219A/E) that were the most predictive for the week 4 virological response, that is, leading to the lowest P value. The 'removing' procedure, starting from a set of mutations retained and removing mutations one by one to find the best combination, provides lower P values than the 'adding' procedure starting with a single mutation and adding mutations one by one. Whatever the set of mutations and the procedure used, the Jonckheere-Terpstra test selects combinations of mutations leading to lower P values than the Kruskal-Wallis test. CONCLUSION: The Jonckheere-Terpstra test for trend is recommended for building a genotypic score when compared with the Kruskal-Wallis. The choice of the selection procedure is discussed here and may be dependent on the objective of the score. PMID- 16038474 TI - Are adverse events of nevirapine and efavirenz related to plasma concentrations? AB - OBJECTIVE: The relationships between adverse events (AEs) and plasma concentrations of nevirapine (NVP) and efavirenz (EFV) were investigated as part of the large, international, randomized 2NN study. METHODS: Treatment-naive, HIV 1-infected patients received NVP (once or twice daily), EFV or their combination, each in combination with lamivudine and stavudine. Blood samples were collected on day 3 and weeks 1, 2, 4, 24 and 48. Concentrations of NVP and EFV were quantitatively assessed by a validated HPLC assay. Individual Bayesian estimates of the area under the plasma concentration-time curve over 24 h (AUC24h), and minimum and maximum plasma concentrations (Cmin and Cmax) as measures for drug exposure of NVP and EFV, were generated using a previously developed population pharmacokinetic model. Pharmacokinetic parameters were compared for patients with and without central nervous system (CNS) and psychiatric AEs, hepatic events, liver enzyme elevations (LEEs) and rash. Furthermore, it was investigated whether a clear cut-off for a pharmacokinetic parameter could be identified above which the incidence of AEs was clearly increased. AEs were also related to demographic parameters and baseline characteristics. RESULTS: In total, from 1077 patients, NVP (3024 samples) and EFV (1694 samples) plasma concentrations and AE data (825 observations) were available. For all patients Cmin, Cmax and AUC24h were determined. When corrected for known covariates of gender, CD4 cell count at baseline, region, hepatitis coinfection and possible interactions between these factors, no significant associations between AEs and any tested exposure parameter of NVP was observed. Also, no target Cmin value, above which patients were at increased risk for AEs, could be established. On the other hand, geographical region, hepatitis coinfection, CD4 cell count and gender were found to be significantly related with the incidence of CNS and psychiatric AEs, hepatic events, LEEs and rash during the treatment with NVP. The occurrence of elevated liver enzymes during the first 6 weeks in the EFV-containing arm was significantly (P = 0.036) correlated to the exposure of EFV (Cmin). Only hepatitis coinfection impacted on LEEs during the first 6 weeks of treatment. With an EFV Cmin above 2.18 mg/l during the induction phase, patients were 4.4 (range 1.3-15.5) times more at risk for elevated liver enzymes. No other correlations between AEs and EFV pharmacokinetics or patient characteristics could be identified. CONCLUSIONS: Pharmacokinetic parameters of NVP did not have a relationship to AEs in the 2NN trial when corrected for known covariates. The value of periodical drug monitoring of NVP as a way to prevent toxicity is therefore limited. Treating physicians should instead focus on factors that are more predictive of AEs (gender, CD4 count and hepatitis coinfection). High EFV Cmin levels resulted in elevated liver enzyme values during the first 6 weeks of treatment. Regular measurement of EFV levels and liver enzymes at the start of therapy may therefore be advised. PMID- 16038475 TI - Evolution of hepatitis C virus quasispecies during therapy with IL2 combinated to alpha interferon and ribavirin. AB - This study analyses the impact of interleukin 2 (IL2) combined with alpha interferon (IFN-alpha) and ribavirin on the heterogeneity of hepatitis C virus (HCV). We studied 10 patients who took part in a clinical trial that assessed the effects of retreatment with IL2, IFN-alpha and ribavirin in patients who failed to clear the virus after a previous bitherapy. The heterogeneity of HCV quasispecies was assessed by cloning and sequencing the hypervariable region 1 (HVR1) in samples obtained at baseline (W0), after 12 weeks of treatment with IFN alpha and ribavirin (W12), after a cycle of administration of IL2 in combination with the classical bitherapy (W21 and W24) in the eight patients who failed to clear the virus under treatment. The mean viral load at W21 and at W24 was not different from that at W12. The heterogeneity of HVR1 quasispecies after the administration of IL2 was not different from that at baseline or after 12 weeks of bitherapy. Furthermore, the proportion of nonsynonymous substitutions was unchanged after the IL2 cycles. Thus, the efficacy of the tritherapy with IL2, IFN-alpha and ribavirin is similar to that of the classical bitherapy. Treatment with IL2 in combination with IFN-alpha and ribavirin had no effect on the selective pressure on HCV quasispecies. IL2 is not the best option to treat hepatitis C. PMID- 16038476 TI - Early virological failure after tenofovir + didanosine + efavirenz combination in HIV-positive patients upon starting antiretroviral therapy. AB - A prospective, randomized pilot trial was conducted in naive patients comparing three different combinations: zidovudine+lamivudine+lopinavir/ritonavir (arm A) versus tenofovir+lamivudine+efavirenz (arm B) versus tenofovir+didanosine+efavirenz (arm C). HIV-RNA slope (days 1, 3, 7, 14 and 28) was slower in arm C with respect to arm B (P < 0.0001). Seven out of eight patients (87.5%) reached undetectable HIV-RNA by week 28 in arm A, 10/10 (100%) in arm B and 6/10 (60%) in arm C. Among arm C patients who failed at week 4, one HIV isolate showed 67N and 219Q, and another one showed 210F and 215D substitutions in the HIV reverse transcriptase gene at baseline, respectively. Non-nucleoside reverse transcriptase inhibitor resistance-related mutations appeared first, followed by 65R mutations in all cases. Efavirenz AUC(0-24) values were lower in arm C with respect to arm B, especially in patients who failed early. A high virological failure rate after tenofovir+didanosine+efavirenz correlated with a slower HIV-RNA decrease and a peculiar accumulation of resistance mutations. A constellation of factors could be correlated with early failure events in patients receiving this combination such as resistance mutations or polymorphisms present at baseline, low CD4+ T cell count or advanced disease and unexpectedly low efavirenz plasma levels. PMID- 16038477 TI - Reverse transcriptase inhibitors alter uncoupling protein-1 and mitochondrial biogenesis in brown adipocytes. AB - OBJECTIVE: Human adipose depots contain remnant brown adipocytes interspersed among white adipocytes, and disturbances of brown with respect to white adipocyte biology have been implicated in highly active antiretroviral therapy (HAART) induced lipomatosis. Brown adipocytes express the uncoupling protein-1 (UCP1) and contain a large number of mitochondria, potential targets of HAART toxicity. The aim of this study was to evaluate the effects of reverse transcriptase inhibitors (RTIs) on primary brown adipocytes differentiated in culture. DESIGN AND METHODS: We analysed the effects of RTIs, nucleoside analogues (NRTIs: stavudine, zidovudine, didanosine and lamivudine) and non-nucleoside analogues (NNRTIs: nevirapine and efavirenz), on differentiation, mitochondrial biogenesis and gene expression in brown adipocytes. RESULTS: None of the NRTIs altered brown adipocyte differentiation whereas NNTRIs had differing effects. Efavirenz blocked lipid deposition and expression of adipose marker genes but nevirapine induced lipid accumulation and adipose gene expression, promoted mitochondrial biogenesis and increased UCP1. Stavudine, zidovudine and didanosine reduced mitochondrial DNA (mtDNA) content. However, mitochondrial genome expression was only impaired in didanosine-treated adipocytes. Stavudine, but not zidovudine, induced expression of the mitochondrial transcription factors and this may explain compensatory mechanisms for the depletion of mtDNA by up-regulating mtDNA transcription. Stavudine caused a specific induction of UCP1 gene expression through direct interaction with a retinoic acid-dependent pathway. CONCLUSIONS: Specific disturbances in brown adipocytes in adipose depots may contribute to HAART-induced lipomatosis. Mitochondrial depletion does not appear to be the only mechanism explaining adverse effects in brown adipocytes because there is evidence of compensatory mechanisms that maintain mtDNA expression, and the expression of the UCP1 gene is specifically altered. PMID- 16038478 TI - Inhibition of replication and infection of severe acute respiratory syndrome associated coronavirus with plasmid-mediated interference RNA. AB - Severe acute respiratory syndrome (SARS) is a newly emerged infectious disease caused by a novel coronavirus (SARS-CoV), which spread to over 30 countries in early 2003. Until recently, no specific vaccines and effective drugs have been available to protect patients from infection by this virus. To exploit a new strategy to fight this disease, we investigated the effect of interference RNA (RNAi) on the virus infection and replication with 3-(4,5-dimethyl-2-thiazolyl) 2,5-diphenyltetrazolium bromide (MTT), plaque-forming, Western-blot and real-time PCR assays. Results showed that the plasmid-derived siRNAs targeting the non structural protein 1 (NSP1) sequence of the SARS-CoV genome could specifically inhibit the expression of the NSP1 sequence and effectively suppress the replication and propagation of SARS-CoV in cultured Vero E6 cell lines. The expression of the Spike and Nucleoprotein genes of SARS-CoV at mRNA and protein levels in small interfering (si)RNA-expressing cells was significantly less than that in controls when analysed with PCR and Western-blot assays, 3 days post SARS CoV infection. Our study provides strong evidence that the NSP1 sequence in the SARS-CoV genome is a valid target for RNAi and the effect of the siRNAs probably mainly resulted from effects on global reduction of subgenome synthesis and subsequent protein expression of SARS-CoV. PMID- 16038479 TI - The clinical utility of using Catrimox-14-treated whole blood in detecting hepatitis C virus RNA. AB - BACKGROUND: Measuring hepatitis C virus (HCV) RNA in serum or plasma may underestimate the true HCV burden. Extracting viral RNA from whole blood (WB) with a cationic surfactant (Catrimox-14) has resulted in HCV RNA concentrations up to 1000-fold higher than from serum or plasma in some studies, but not others. We compared the Catrimox-14 WB assay with a standard serum assay. METHODS: Seventy-two chronic HCV patients received 48 weeks of standard or pegylated interferon alpha-2b and ribavirin therapy. Catrimox-14-treated WB and corresponding serum samples were obtained at baseline and weeks 12, 24, 48 and 72. HCV RNA concentrations from WB and serum were quantified by a previously validated RT-PCR assay. RESULTS: Overall mean (+/- SD) baseline serum log10 HCV RNA concentration was 6.5 ((+/- 0.58) copies/ml. Out of 72 patients, 33 had no detectable virus at 72 weeks. Neither assay detected virus in these patients at 12 weeks and neither WB nor serum assays detected virus at end-of-treatment in the 10 patients who subsequently relapsed at 72 weeks. HCV RNA concentrations from WB and serum assays were linearly correlated (R2 = 0.73; P < 0.001), although mean serum HCV RNA concentrations were 0.5 log10, copies/ml higher in serum than in WB [6.0 (+/- 0.82) vs 5.5 ((+/- 0.84), respectively, P = 0.12]. CONCLUSIONS: Catrimox-14-treated WB assays detect changes in HCV RNA, but do not offer clinical advantage over a conventional serum RT-PCR based assay. PMID- 16038480 TI - Muscle and liver lactate metabolism in HAART-treated and naive HIV-infected patients: the MITOVIR study. AB - OBJECTIVE: To assess the impact of nucleoside analogue reverse transcriptase inhibitor (NRTI) combination therapy on muscle and liver lactate metabolism in HIV-infected patients. METHODS: This cross-sectional study involved HIV-infected patients who were either antiretroviral-naive (Group 1) or were receiving either a stable triple-drug combination including at least one d-drug (zidovudine, zalcitabine, stavudine, didanosine; Group 2) or a backbone of abacavir and lamivudine (Group 3). Lactataemia was measured at rest. Muscle lactate metabolism was assessed during a standardized exercise test and liver lactate metabolism during intravenous lactate infusion. Mitochondrial DNA was quantified in peripheral blood mononuclear cells. RESULTS: A total of 65 patients were enrolled (16, 31 and 18 patients in Group 1, Group 2 and Group 3, respectively). None of the patients had symptoms of hyperlactataemia. Patients in Group 3 had received d drugs for a median of seven years before switching to abacavir and lamivudine. Median baseline lactataemia, although within the normal range, was significantly higher in both treatment groups than in the naive patients (Group 2: 1.4, Group 3: 1.5, and Group 1: 1.0 mmol/l, P = 0.005). Muscle lactate clearance was significantly lower in both treatment groups than in naive patients (Group 2: 1.6, Group 3: 1.8, and Group 1: 2.1, P = 0.01). Lactate liver metabolism and mitochondrial DNA levels did not differ among the three groups. CONCLUSIONS: In HIV-infected patients without symptomatic hyperlactataemia, all NRTI-containing HAART regimens appear to cause muscle mitochondrial damage but to spare the liver. Absence of difference between Group 2 and Group 3 raises questions about the potential reversibility of muscle mitochondrial dysfunction, and/or the ability of abacavir and lamivudine to induce such mitochondrial damage. PMID- 16038481 TI - Interactive selective pressures of HLA-restricted immune responses and antiretroviral drugs on HIV-1. AB - HIV-specific cytotoxic T lymphocyte (CTL) responses mediated by human leukocyte antigen (HLA) recognition and antiretroviral drugs exert selection pressure on HIV-1 in vivo. The selection of CTL escape mutations strongly underpins the failure of CTL control in most untreated infections whilst drug-resistance mutations predict failure of drug control. These two evolutionary forces share common target residues in HIV-1 at which their selection effects could be synergistic or antagonistic, such that the propensity to develop drug resistance and virological treatment failure may be influenced by HLA type. We examined HIV 1 reverse transcriptase (RT) and protease sequences in a large clinical observational cohort of 487 HIV-infected individuals and found evidence of site specific interactions between specific antiretroviral drug exposures, HLA alleles and HIV sequence diversity at population level. Such interactions may have general and specific implications for explaining in vivo/in vitro discordance of drug resistance, host-specific susceptibility to drug resistance, individualization of therapy and therapeutic vaccine design. PMID- 16038482 TI - Mitochondrial DNA depletion in HIV-infected patients is more pronounced with chronic hepatitis C and enhanced following treatment with pegylated interferon plus ribavirin. AB - BACKGROUND: Mitochondrial DNA (mtDNA) damage seems to be responsible for many of the toxicities associated with the long-term use of nucleoside analogues in HIV infected patients. These adverse effects, mainly lipoatrophy, seem to be even more pronounced in subjects with hepatitis C virus (HCV) co-infection. However, there is no information about a possible additive effect of HCV on mtDNA depletion nor about the impact of ribavirin use in HIV/HCV-coinfected individuals. PATIENTS AND METHODS: mtDNA was measured in peripheral blood mononuclear cells (PBMC) collected from 192 individuals classified into 4 groups: HIV-neg/HCV-neg (control group, n = 11), HIV-pos/HCV-neg (56), HIV-neg/HCV-pos (18) and HIV-pos/HCV-pos (107). A duplex real-time NASBA assay was used to quantify mtDNA on maximal platelet-depleted specimens and all experiments were run in duplicate. The mtDNA copy number per cell was estimated taking as reference the nuclear DNA copy number. RESULTS: The mean mtDNA values in the control group was 757 copies/cell, while it was 428, 349 and 296 for HIV-pos, HCV pos and HIV/HCV-coinfected individuals, respectively (P < 0.001 for all groups relative to the control group). No significant differences were observed when comparing patients with HIV or HCV infections alone, but coinfected individuals showed a lower mtDNA copy number than patients infected with HIV (P < 0.001) or with HCV (P = 0.089). In a subset of 18 patients with HIV/HCV-coinfection, treatment with pegylated interferon plus ribavirin produced a further reduction in mtDNA (mean value, 189 copies/cell; P = 0.009). CONCLUSIONS: HIV and HCV may independently cause mtDNA depletion in PBMC. Coinfection may result in more pronounced mtDNA depletion. The administration of interferon plus ribavirin may further enhance mtDNA depletion. These findings may explain the greater risk of lipoatrophy of antiretroviral therapy in HIV-infected patients with HCV coinfection and why anti-HCV therapy may aggravate this effect. PMID- 16038483 TI - Impact of newly available drugs on clinical progression in patients with virological failure after exposure to three classes of antiretrovirals. AB - OBJECTIVE: To study the prognosis of HIV-infected patients with virological failure after exposure to three classes of antiretroviral drugs (ARVs). DESIGN: Cohort study. SETTING: French Hospital Database on HIV. PATIENTS: Patients previously exposed to at least two nucleoside reverse transcriptase inhibitors (NRTIs), two protease inhibitors and one non-NRTI, with viral load (/L) values of > 5000 copies/ml after the exposure criteria were met and a new treatment initiated between 1998 and 2001 with VL > 5000 copies/ml. MAIN OUTCOME MEASURES: Risk of new AIDS-defining-events (ADEs) or death from first introduction of a drug never used before occurring between 1998 and 2001 defined as baseline. RESULTS: The main baseline characteristics of the 1092 patients were: previous ADE in 49% of cases, median CD4 cell count 181 microl, median VL 4.9 log10 copies/ml, median duration of ARV therapy 5.0 years and previous exposure to a median of nine ARVs. The crude progression rates were 20.1/100 patient-years among patients included in 1998, 15.1 in 1999, 11.1 in 2000 and 8.6 in 2001. After adjustment for baseline characteristics, the calendar year of inclusion was associated with the risk of clinical progression (P < 0.001). When the types of newly available drugs used at baseline or during follow-up were introduced into the model, year of inclusion was no longer associated with the risk of clinical progression (P = 0.42), while exposure to amprenavir/r, lopinavir/r, abacavir or tenofovir was associated with a lower risk. CONCLUSIONS: The clinical prognosis of heavily pretreated patients experiencing virological failure improved between 1998 and 2001, mainly thanks to the use of newly available drugs with more favourable resistance profiles. PMID- 16038484 TI - Facial fat volume in HIV-infected patients with lipoatrophy. AB - BACKGROUND: The volume and anatomical distribution of facial fat depletion in HIV related lipoatrophy have not been properly quantified. We aimed to determine the extent and distribution of facial fat loss in HIV-infected patients with lipoatrophy and whether this differs from wasting. DESIGN: We studied HIV infected patients with clinically defined moderate to severe lipoatrophy without wasting (n = 15), clinically defined wasting (> 10% weight loss and recent opportunistic infection) with no previous reported lipodystrophy (n = 15), and controls without lipodystrophy or wasting (n = 15). Facial MRI scans were performed, and areas of fat were manually traced bilaterally in all individual image slices and summed to calculate volume in anatomically defined regions of interest. RESULTS: Patients with lipoatrophy had lower fat volumes in the temporal region (8.9 +/- 9.4 vs 20.5 +/- 7.2 ml; P < 0.001), cheek region (25.6 +/- 24.9 vs 55.5 +/- 15.0 ml; P < 0.001), periorbital region (1.9 +/- 1.0 vs 2.7 +/- 1.0 ml; P = 0.09) and buccal fat pad (13.4 +/- 9.4 vs 21.8 +/- 9.8 ml; P = 0.030) compared with controls. Patients with wasting had temporal, cheek, periorbital and buccal fat pad volumes (10.4 +/- 6.7 ml, 34.0 +/- 14.8 ml, 1.4 +/ 1.1 ml and 13.1 +/- 4.6 ml respectively) that were lower than controls (all P < 0.01) but similar to lipoatrophy patients (all P > 0.5). CONCLUSIONS: Facial fat depletion in lipoatrophy is substantial (approximately 50% volume loss) and involves superficial and deep fat (buccal fat pad). The distribution and volume of fat change is similar to that seen in wasting. Given the extent of the changes, complete surgical correction is unlikely to be possible and hence emphasis should be placed on prevention of lipoatrophy [corrected] PMID- 16038485 TI - Poly(amide amine) dendrimer with naphthyl units as a fluorescent chemosensor for metal ions. AB - Poly(amide amine) dendrimer with naphthyl units (N8) as a fluorescent chemosensor for metal ions was synthesized. We investigated the metal ion recognition of N8. Large changes in the fluorescence spectra of N8 were observed upon the addition of cadmium and zinc ions. PMID- 16038486 TI - Radiative auger effect and extended X-ray emission fine structure (EXEFS). AB - Radiative Auger spectra are weak X-ray emission spectra near the characteristic X ray lines. Radiative Auger process is an intrinsic energy-loss process in an atom when a characteristic X-ray photon is emitted, due to an atomic many-body effect. The energy loss spectra correspond to the unoccupied conduction band structure of materials. Therefore the radiative Auger effect is an alternative tool to the X ray absorption spectroscopy such as EXAFS (Extended X-ray Absorption Fine Structure) and XANES (X-ray Absorption Near Edge Structure), and thus it is named EXEFS (Extended X-ray Emission Fine Structure). By the use of a commercially available X-ray fluorescence spectrometer or an electron probe microanalyzer (EPMA), which are frequently used in materials industries, we can obtain an EXEFS spectrum within 20 min. The radiative Auger effect, as an example, demonstrates that the study on atomic many-body effects has become a powerful tool for crystal and electronic structure characterizations. The EXEFS method has already been used in many industries in Japan. Reviews about the applications and basic study results on the radiative Auger effect are reported in this paper. PMID- 16038487 TI - Quality assurance challenges in X-ray emission based analyses, the advantage of digital signal processing. AB - There is a large scatter in the results of X-ray analysis with solid-state detectors suggesting methodological origin. In order to improve the methodology, detector response functions have been investigated by many researchers and analysts. This was necessary as the departure of the response function of some detector-signal processing electronics from the normally assumed Gaussian line shape can exceed 100% in area. Several detector models have been proposed to improve understanding and establish a firm basis for quantitative work. After reviewing some contradictory results, we describe a signal processor that offers quality assurance, by producing two spectra for each measurement. One is the normal spectrum of accepted events, while the second spectrum contains all of the rejected events. For each measurement, therefore, all X-ray events are recorded, enabling quality control. In addition to this improvement, the digital signal processor of Cambridge Scientific, Canada, delivers a high throughput rate, excellent resolution, decreased low energy tailing and a line shape justified by the physics of the detector. Comparative measurements are presented to demonstrate the improved rejection of background from gamma rays as well as a significant improvement in pile-up recognition. The rejected events spectrum gives insight into the origin of the response function, which suggests that the flat plateau of the frequently used Hypermet function, normally attributed to detector dead layers, originates from pile-up with the low energy noise events. A detailed analysis demonstrates how the relative intensities of the X-ray lines can change in a varying noise environment, thus potentially explaining the unacceptable large scatter in the experimental data currently found in the literature. The comparison of the accepted and rejected events adds the possibility of monitoring the electronic efficiency of signal recognition that has generally been ignored in this field. PMID- 16038488 TI - Study of heavy metals in wild edible mushrooms under different pollution conditions by X-ray fluorescence spectrometry. AB - In this work we studied and compared the metal uptake in edible mushrooms (Lepiota procera, Boletus badius, Boletus edulis, Tricholoma equestry, Lactarius deliciosus, Cantarelus tubalformis and Cantarelus edulis), relative to sampling sites submitted to different pollution conditions: car traffic, soil pollution due to pesticides and fertilizers used in old vineyards, and incineration of hospital waste. Soil was also collected in some places, and its content was correlated to the corresponding one in some mushrooms species. All samples, without any chemical treatment, were analyzed by an X-ray fluorescence set-up. This technique is based on a monochromatic X-ray beam ionizing the atoms of the sample. Following this ionization, the emitted radiation is characteristic of the element, allowing its identification and quantification. Vineyards are normally submitted to very high amounts of sulfating, containing high copper concentrations. This metal is accumulated on the soil, and can be up-taken by vegetation. Very high levels of Fe and Cu were found in Lepiota procera species in old vineyards. Zinc was found to be always higher than Cu by factors ranging from 1.5 to 8 in clean wood taken as a reference for the whole analyzed species, while in old vineyards the ratio Zn/Cu reach 0.25 for Lepiota procera. This is correlated to the soil content for both elements. In addition, pollution induced by car traffic was checked in some samples, collected in the proximity of highways. Pb was the main contaminant in these areas, and presenting values 10 times higher than the corresponding ones in sites not submitted to pollution, for some species. Mushrooms contamination due to incineration of hospital waste was also studied, but we did not observe any contamination involving heavy metals in the several analyzed species around these areas. This is in agreement with what was expected, taking into account that hospital waste is mostly organic and, in principle, no heavy metals would be observed. PMID- 16038489 TI - Soil characterization by energy dispersive X-ray fluorescence: sampling strategy for in situ analysis. AB - This work describes a sampling strategy that will allow the use of portable EDXRF (energy dispersive X-ray fluorescence) instruments for "in situ" soil analysis. The methodology covers a general approach to planning field investigations for any type of environmental studies and it was applied for a soil characterization study in the zone of Campana, Argentina, by evaluating data coming from an EDXRF spectrometer with a radioisotope excitation source. Simulating non-treated sampled as "in situ" samples and a soil characterization for Campana area was intended. "In situ" EDXRF methodology is a powerful analytical modality with the advantage of providing data immediately, allowing a fast general screening of the soil composition. PMID- 16038490 TI - Effect of energy dependence of primary beam divergence on the X-ray standing wave characterization of layered materials. AB - Incident primary beam divergence is a source of systematic error in X-ray standing wave (XSW) characterization of single and multilayer thin films. Primary beam divergence significantly alters the XSW profile of a layered material and can lead to large errors when used with higher excitation energies. The present study suggests that when one uses Mo-Kalpha excitation, the primary beam divergence should be in range of 0.005(0). On the other hand, in the case of Cu Kalpha excitation, primary beam divergence can be relaxed up to 0.01(0). PMID- 16038491 TI - Elucidating a particulate matter deposition episode by combining scanning electron microscopy and X-ray fluorescence spectrometry. AB - Multielemental composition and morphology of particulate matter samples were examined to detect the presence of two potential responsible pollutant emitters at four sites impacted during a pollution episode in the City of Campana, Argentina. Coke and smoke black are the main constituents associated to the industrial plants that were considered, a priori, as responsible pollutant emitters. Wavelength dispersive X-ray fluorescence (WDXRF) was employed for the analysis of metals in samples coming from both, suspected sources and four sites. On the basis of multielemental composition profiles, a screening analysis was undertaken for exploring similarities among sources and sample sites adopting the average concentration profile of the crustal rock as soil surrogate. Particle morphology was studied by scanning electron microscopy (SEM) in source samples and in those environmental samples selected through the screening analysis. Two types of collected samples were analyzed: 1) those with composition profiles closer to the potential sources and 2) those closer to the cluster rock. This strategy was adequate to identify the responsible source of the contamination episode. PMID- 16038492 TI - Determination of chemical form of antimony in contaminated soil around a smelter using X-ray absorption fine structure. AB - Only limited information is available about the behavior of antimony (Sb) in contaminated soils. However, understanding the behavior of Sb in contaminated soils is important, because the toxicity or solubility of this element depends on its chemical state. In this study, we investigated the levels of Sb and the chemical forms of Sb in the soil around a smelter using X-ray absorption fine structure (XAFS) spectra. The highest Sb concentration in the contaminated soil was 2900 mg/kg dry soil. According to Sb-K edge X-ray absorption near edge (XANE) spectra, the Sb in the soil was in the form of Sb(V) compounds. The similarity of extended XAFS (EXAFS) spectra suggests that Sb speciation was independent of the sampling site, which indicates that Sb or Sb2O3 emitted from the smelter was converted into Sb(V) compounds in the soil. PMID- 16038493 TI - Evaluation of a quantitative method for trace impurities in arsenous acids by synchrotron radiation X-ray fluorescence spectrometry and inductively coupled plasma-atomic emission spectrometry. AB - Thirteen arsenous acid samples of known origins and refining methods were collected. Each sample was subjected to quantitative analysis of any impurity elements present using synchrotron radiation X-ray fluorescence spectrometry (SR XRF) and inductively coupled plasma-atomic emission spectrometry (ICP-AES). The trace elements selected were Sn, Sb and Bi for the reasons that they were considered not to be changed by their circumstances and that they showed high sensitivity to SR-XRF. These results obtained by both methods were compared and the correlation between these two methods was determined. The quantification of trace impurities obtained by SR-XRF using As as an internal standard showed good agreement with the results obtained by ICP-AES. The discrimination of refining method became possible by the comparison of these impurities' contents measured with non-destructive SR-XRF using several arsenous acid particles. PMID- 16038494 TI - Excitation energy dependence for the Li 1s X-ray photoelectron spectra of LiMn2O4. AB - The Li 1s XPS (X-ray Photoelectron Spectroscopy) spectra of LiMn2O4, which is one of the major positive-electrode materials in lithium-ion rechargeable batteries, and MnO2 as a reference material, were measured by a laboratory-type XPS spectrometer. The Li 1s peak was not observed in the spectra excited by the Mg Kalpha line (1253.6 eV), because the Li 1s peak overlapped the background of the Mn 3p peak of LiMn2O4. The photoionization cross section of Mn 3p was larger than that of Li 1s for Mg Kalpha excitation. Therefore, the XPS measurement of LiMn2O4 by soft X-ray synchrotron excitation was carried out at beamline BL-7B on NewSUBARU synchrotron facility. Excitation energies of 110, 120, 130, 140, 150 and 151.4 eV were selected. The Li 1s peak was clearly observed in these XPS spectra. In order to investigate the excitation energy dependence, the area ratio of the Li 1s and Mn 3p peaks in the XPS spectra was plotted against the excitation energy. As a result, when the excitation energy was 110 eV, the area ratio had the maximum value. PMID- 16038495 TI - X-ray absorption spectral analysis with a 9 V battery X-ray generator. AB - X-ray absorption spectral (XAS) analysis is performed with a combination of a 9 V dry electric battery X-ray generator and a portable Si PIN X-ray detector. The calcium K edge (4.0 keV) in paper is measured with a grazing incidence geometry, which suppressed the artifact due to the Kalpha X-ray fluorescence peak at 3.5 keV. The 9 V dry battery X-ray emitter is useful for portable XAS measurements. PMID- 16038496 TI - Nondestructive analysis of silver in gold foil using synchrotron radiation X-ray fluorescence spectrometry. AB - Small particles of gold foil detached from an indoor decoration might be important evidence to associate a suspect with a crime scene. We have investigated the application of elemental analysis using synchrotron radiation X ray fluorescence spectrometry to discriminate small particles of gold foil. Eight kinds of gold foil samples collected in Japan were used in the experiments. As a result of synchrotron radiation X-ray fluorescence spectrometry, only two elements, gold and silver, were detected from all gold foil samples. The intensity ratios of AgKalpha/AuLalpha showed good correlation with the content ratios of Ag/Au. The variation of intensity ratio within a same sample was sufficiently small compared with those of different samples. Therefore the comparison of this intensity ratio can be an effective method to discriminate small particles originating from different types of gold foil. PMID- 16038497 TI - The effect of UV irradiation on the reduction of Au(III) ions adsorbed on manganese dioxide. AB - The effect of UV (ultraviolet) irradiation on the adsorption of Au(III) ions on manganese dioxide and their reduction to Au(0) (gold with 0 valence state) was investigated using XPS (X-ray photoelectron spectroscopy) and 197Au Mossbauer spectroscopy. The UV irradiation accelerated the adsorption and the reduction. From the fact that the proportion of Au(0) estimated from Au 4f XPS spectra for surface analysis was significantly smaller than that from 197Au Mossbauer spectra for bulk analysis, we deduced that Au(0) was interpenetrated to the inside of manganese dioxide (into deeper places than about 30 A) where XPS is impossible to detect. The content of surface hydroxyl groups on manganese dioxide also increased due to the UV irradiation. The relationship between the charge in the content of hydroxyl groups and the interpenetration of Au(0) is discussed. PMID- 16038498 TI - Rapid identification of inorganic salts using energy dispersive X-ray fluorescence. AB - A simple method for the rapid identification of inorganic salts used as raw materials during food production has been established using energy dispersive X ray fluorescence (EDXRF). Intensities of Na, Mg, P, S, Cl, K, Ca, Mn, Fe, Cu, Zn Kalpha peaks have been measured. The treatment and comparison of the respective intensities allowed a rapid identification of inorganic salts. Two different methods of data treatment have been tested: K nearest neighbours (KNN) and centroids. Using cross validation on a database containing 140 samples representing 31 classes, a total of 98% of samples were well identified. PMID- 16038499 TI - Grazing-exit and micro X-ray fluorescence analyses for chemical microchips. AB - Grazing-exit x-ray fluorescence (GE-XRF) and micro x-ray fluorescence (micro-XRF) methods were applied to chemical microchips as a detection method. Since an energy-dispersive x-ray detector was used, the simultaneous detection of multiple elements was possible. An analyzing region was especially designed on the microchip so that a sample solution could be dried and concentrated in a suitable area corresponding to the size of the primary x-ray beam. Finally, it was confirmed that both analytical methods could be combined well for use with a microchip. In GE-XRF, the background intensity in the XRF spectrum was reduced at grazing-exit angles. In addition, a good relationship between the x-ray fluorescence intensities and the concentrations of standard solutions that were introduced into the microchip was obtained. This indicates that the GE-XRF method is feasible for trace elemental analysis in chemical microchip systems. In micro XRF, an attempt was made to concentrate and dry the analyte within a small analyzing region. The preliminary results indicated that the micro-XRF method could be applied for the analysis of microchips. PMID- 16038500 TI - Pre-edge features of Ti K-edge X-ray absorption near-edge structure for the local structure of sol-gel titanium oxides. AB - Titanium K-edge X-ray absorption near-edge structure (XANES) spectroscopy is used to examine the local Ti environments in the sol, gel, and xerogels of titanium oxide prepared by a sol-gel method. The xerogels were prepared by heat treatment at 200, 300, 400, 500, and 600 degrees C (denoted xero-200, xero-300, xero-400, xero-500, and xero-600), and the xerogels were doped with Fe(II), Ni(II), and Cu(II) ions (xero-Fe, xero-Ni, and xero-Cu). The local structures of the samples are compared using the pre- and post-edge features of the XANES spectra. Further analysis of the pre-edge features using Gaussian functions provides the energy and height of each peak, and the intensity ratios between peaks. Based on the resulting data, the samples can be classified into three groups: a weak Ti-Ti interaction group, an anatase-like structure group, and an anatase group. The sol, gel, xerogel, and xero-Ni are in the weak Ti-Ti interaction group, and the xero-200, xero-Fe, and xero-Cu are in the anatase-like structure group. The remaining samples are in the anatase group. PMID- 16038501 TI - Numerical simulation of C/C/C planar X-ray waveguides. AB - A numerical simulation was done to check the possibility of using planar C/C/C multilayers with density contrast 0.2 and 0.7 g/cm3 as an X-ray waveguide. After an optimization procedure, suitable waveguide layer thicknesses were found which provide a high degree of resonant standing wave field intensity enhancement in the core layer at incident beam energy of 13 keV. The obtained results were compared with those of the Mo/Be/Mo waveguide, whose high waveguiding capability at the same energy value was reported in the 1990s. The comparison shows that standing wave field intensity resonant enhancement provided by C/C/C planar multilayers is very high and, consequently, a guided beam can be well detected. PMID- 16038502 TI - X-ray fluorescence analysis of rare earth elements in rocks using low dilution glass beads. AB - Major and trace elements (Na, Mg, Al, Si, P, K, Ca, Ti, Mn, Fe, Rb, Sr, Y, Zr, La, Ce, Pr, Nd, Sm, Gd, Dy, Th and U) in igneous rocks were assayed with fused lithium borate glass beads using X-ray fluorescence spectrometry. Low dilution glass beads, which had a 1:1 sample-to-flux ratio, were prepared for determination of rare earth elements. Complete vitrification of 1:1 mixture required heating twice at 1200 degrees C with agitation. Extra pure reagents containing determinants were used for calibrating standards instead of the rock standard. The calibration curves of the 23 elements showed good linearity. Furthermore, the lower limits of detection corresponding to three times the standard deviation for blank measurements were 26 mass ppm for Na2O, 6.7 for MgO, 4.5 for Al2O3, 4.5 for SiO2, 18 for P2O5, 1.1 for K2O, 4.0 for CaO, 3.9 for TiO2, 1.6 for MnO, 0.8 for Fe2O3, 0.5 for Rb, 0.2 for Sr, 0.4 for Y, 0.5 for Zr, 3.3 for La, 6.5 for Ce, 2.7 for Pr, 2.1 for Nd, 1.7 for Sm, 0.7 for Gd, 2.7 for Dy, 0.5 for Th, and 0.6 for U. Using the present method, we determined the contents of these 23 elements in four rhyolitic and granitic rocks from Japan. PMID- 16038503 TI - Complexing membrane for uranium detection by total reflection X-ray fluorescence. AB - Selective membranes containing a few micrograms of various complexing reagents in a polyvinyl chloride (PVC) matrix were produced in order to determine low uranium concentrations in water. The membranes were produced on the surfaces of quartz glasses, and immersed in water solutions containing uranium salts (5 - 50 ng/mL). By the end of the equilibration time, the membranes were left to dry and analyzed based on the total reflection X-ray fluorescence. The effects of the complexing agents and their mass proportions added in the membranes were studied. The combination of the complexing reagents dithizone and thiourea gave the best result. The minimum detection limit was measured to be equal to 0.8 ng/mL. PMID- 16038504 TI - Determination of trace elements in sediment reference materials by monochromatic X-ray excitation X-ray fluorescence spectrometry. AB - A new reliable analytical method, "Monochromatic X-ray Excitation X-ray Fluorescence Spectrometry", has been proposed. For validating the method, trace elements in sediment certified reference materials were determined. In the method X-ray fluorescence spectra are measured for specimens and pure metals; in addition the mass-attenuation coefficients of the specimens for various X-ray wavelengths are also measured. The data are analyzed by the fundamental parameter method and the uncertainty of the analysis is evaluated. The obtained results were in satisfactory agreement with the certified values within their uncertainties. This method will be applicable to the certification of reference materials, in the field of which reliable results with uncertainty statements are required. PMID- 16038505 TI - X-ray powder diffraction patterns for certain beta-lactam, tetracycline and macrolide antibiotic drugs. AB - X-ray powder diffraction (XRD) data for eight beta-lactam viz., ampicillin sodium, ampicillin trihydrate, penicillin G procaine, benzathine penicillin, benzyl penicillin sodium, cefalexin, cefotaxime sodium and ceftriaxone sodium; three tetracyclines viz., doxycycline hydrochloride, oxytetracycline dihydrate and tetracycline hydrochloride; and two macrolide viz., azithromycin and erythromycin estolate antibiotic drugs were obtained using a powder diffractometer. The drugs were scanned from Bragg angles (2theta) of 10 degrees to 70 degrees. The obtained data were tabulated in terms of the lattice spacing (A) and relative line intensities (I/I(I)). This new information may be useful for identifying these drugs from confiscated materials, which has been frequently encountered in forensic laboratories. PMID- 16038506 TI - Investigation of individual micrometer-size Kosa particle with on-site combination of electron microscope and synchrotron X-ray microscope. AB - The Kosa (yellow sand) aerosol affects the global environment as well as human health because it migrates from the interior of China to other areas, absorbing various atmospheric elements. Investigation into individual Kosa aerosol particles, which are submicroscopic to several tens of micrometers in diameter, is required to resolving the issue. We installed a scanning electron microscope (SEM) equipped with an energy dispersive X-ray spectrometer (EDX) on a synchrotron radiation (SR) beam line and introduced the SR beam into the SEM chamber for combinatorial application of SEM-EDX and SR X-ray fluorescence (SR XRF) spectrometry to individual particles. It should be noted that detailed topographic observation by SEM and sensitive elemental analysis by SR-XRF, both crucial for individual particle measurement but which previously had to be carried out separately, were jointly performed inside the SEM chamber in this setup. Here, we show that SR-XRF results, in conjunction with SEM images, contributed toward resolving individual particle dispositions. Atmospheric sulfur primarily adheres to calcium in the aerosol particles and the particle surface roughens as a consequence of the chemical reaction between the two elements. PMID- 16038507 TI - Grazing incidence-X-ray fluorescence spectrometry for the compositional analysis of nanometer-thin high-kappa dielectric HfO2 layers. AB - In future microelectronic devices, SiO2 as a gate dielectric material will be replaced by materials with a higher dielectric constant. One such candidate material is HfO2. Thin layers are typically deposited from ligand-containing precursors in chemical vapor deposition (CVD) processes. In the atomic layer deposition (ALD) of HfO2, these precursors are often HfCl4 and H2O. Obviously, the material properties of the deposited films will be affected by residual ligands from the precursors. In this paper, we evaluate the use of grazing incidence--and total reflection-X-ray fluorescence spectrometry (GI-XRF and TXRF) for Cl trace analysis in nanometer-thin HfO2 films deposited using ALD. First, the results from different X-ray analysis approaches for the determination of Hf coverage are compared with the results from Rutherford backscattering spectrometry (RBS). Next, we discuss the selection of an appropriate X-ray excitation source for the analysis of traces within the high-kappa: layers. Finally, we combine both in a study on the accuracy of Cl determinations in HfO2 layers. PMID- 16038508 TI - Simultaneous determination of cobalt, copper and zinc by energy dispersive X-ray fluorescence spectrometry after preconcentration on PAR-loaded ion-exchange resin. AB - A sensitive method for the preconcentration and determination of trace amounts of Co, Cu and Zn by energy-dispersive X-ray fluorescence spectrometry (EDXRF) has been developed. The method is based on the fact that 4-(2-pyridylazo)-resorcinol (PAR) loaded Dowex anion-exchange resin (PAR-resin) can effectively adsorb Co, Cu and Zn at pH 9.0 to form PAR-metal complexes. The detection limits for Co, Cu and Zn were 1.53, 0.31 and 0.21 ppb, respectively. The precisions for five replicate measurements of the three metals were 3.4, 2.7 and 2.1% RSD, and the calibration curves were linear up to 75 microg with correlation coefficients of 0.9975, 0.9980 and 0.9985, respectively. The method was successfully applied for the simultaneous determination of Co, Cu and Zn in seawater samples at ppb levels. PMID- 16038509 TI - Forensic discrimination of sheet glass by a refractive-index measurement and elemental analysis with synchrotron radiation X-ray fluorescence spectrometry. AB - Measurements of the refractive index (RI) and elemental analysis using synchrotron radiation X-ray fluorescence spectrometry (SR-XRF) were applied to the forensic discrimination of sheet-glass samples from different origins. The refractive index was calculated from the matching temperature at which the glass fragments became invisible in silicone oil. Fragments smaller than 1 mm in maximum diameter were taken from each of 11 sheet glasses and subjected to analysis by SR-XRF. The XRF spectrum of these samples indicated that a comparison of 6 elements (Ca, Fe, Sr, Zr, Ba and Ce) was useful for the discrimination of sheet glasses. Cluster analysis was performed using 33 sets of SR-XRF data obtained by triplicate measurements for the 11 glasses. Comparing 528 pairs among 33 samples, 515 pairs could be correctly discriminated. The number of indistinguishable pairs could be reduced from 36 to 4 by comparing the SR-XRF data. Elemental analysis by SR-XRF could provide small glass fragments with a more evidential value than the solely measurement of only RI, through a significant improvement of the discrimination capability. PMID- 16038510 TI - Valence determination of manganese in battery cathode materials by high resolution Mn Kalpha1 spectra. AB - High-resolution the Mn Kalpha1 X-ray fluorescence spectra (HRXRF) were measured for a variety of manganese compounds, the oxidation number (valence) of which was from II to VII. Plots of the valence against the full width at half maximum (FWHM) and the chemical shift of the Kalpha1 X-ray fluorescence spectra give a curve and a liniar relation, respectively. The coefficient of correlation (R2) for the latter plot was 0.989 between valency II and VII. More excellent linearlity from III to IV was obtained with R2 = 0.995, which enabled us to determine the oxidation number of manganese atom in a nickel ion doped spinel structure, LiMn2O4 (LiNixMn(2-x)O4, with 0.2 < x < 0.42). From the valence of manganese ion obtained from HRXRF, the oxidation number of nickel ion is concluded to be divalent. PMID- 16038511 TI - Chemical effects in high-resolution nickel Kalpha X-ray fluorescence spectra. AB - The nickel Kalpha spectra of oxides, halides (NiF2, K2NiF6, NiCl2, NiBr2), complex compounds, and metal are measured with two different double-crystal X-ray fluorescence spectrometers. The peak shifts and line width changes due to the changes in the chemical states are reported. High reproducibility has been shown for the chemical shift and line width measurements. The DV-Xalpha molecular orbital calculation at the ground and the 1s(-1) hole states was performed to prove that the chemical effect originats from the charge-transfer mechanism. PMID- 16038512 TI - Test method for the presence or absence of Pb in electrical components using energy-dispersive micro X-ray fluorescence. AB - The micro-EDXRF (energy dispersive X-ray fluorescence) method was applied to the screening of Pb in micrometer-area samples, such as a Cu contact in electrical components that had been coated by Pb-free Sn-Ag-Cu solder. The reliability of the screening method was evaluated by a comparison with a scanning electron microscope (SEM) observation and a precious chemical analysis method of inductively coupled plasma mass spectrometry (ICP-MS). Some factors that affect the testing reliability, such as the thickness of the solder, the segregation of Pb and Ag, etc. were found by SEM observations. By adjusting some calculation parameters, screening of the micrometer area (0.1 mm) was performed using the fundamental parameter (FP) method for a thin film in conjunction with micro EDXRF. The measurement error ranged by 25% for the thin film-FP method. The resulted detection limit was 0.04 wt% for Pb, depending on the solder thickness. This method can be successively applied for quality control to check the purity of a Pb-free Sn-Ag-Cu solder coating in electrical components. PMID- 16038513 TI - Environmental trace-element analysis using a benchtop total reflection X-ray fluorescence spectrometer. AB - Total reflection X-ray fluorescence (TXRF) analysis is an established technique for trace-element analysis in various types of samples. Though expensive large scale systems restricted the applications in the past, in this study the capability of a benchtop system for trace elemental analysis is reported. The suitability of this system for the mobile on-site analysis of heavy metal contaminated soils and sediments is reported as well as the possibilities and restrictions of TXRF for additional applications, including trace-element analysis of water, glass and biological samples. PMID- 16038514 TI - Strong X-ray emission from electrified insulators. AB - Terasawa reported strong X-ray emission from charged-up insulators, and proposed an X-ray production device only using the electrification. We constructed a similar device and studied the conditions of X-ray emission. It was shown that X rays could be produced without supplying electrons from a filament. PMID- 16038515 TI - A simple X-ray emitter. AB - A compact X-ray emission instrument is made, and the X-ray spectra are measured by changing the applied electric potential. Strong soft X-rays are observed when evacuating roughly and applying a high voltage to an insulator settled in this device. The X-ray intensity is higher as the applied voltage is increased. A light-emitting phenomenon is observed when this device emits X-rays. The present X-ray emitter is made of a small cylinder with a radius of 20 mm and a height of 50 mm. This X-ray generator has a potential to be used as an X-ray source in an X ray fluorescence spectrometer. PMID- 16038516 TI - The role of trace metallic elements in neurodegenerative disorders: quantitative analysis using XRF and XANES spectroscopy. AB - The present paper focuses on the analysis of trace metallic elements and their role in neurodegenerative disorders. The use of synchrotron radiation microbeams allows investigation of pathological tissues from Alzheimer's disease, Parkinson's disease and Amyotrophic lateral sclerosis cases in a nondestructive manner and at cellular level. By employing X-ray absorption near edge structure (XANES) technique, the chemical state of the investigated elements can be determined, while energy-selective X-ray fluorescence spectroscopy provides the spatial distribution of each element in each oxidative state selectively. The investigated tissues (derived from human, monkey and mouse specimens) show distinct imbalances of metallic elements such as Zn and Cu as well as Fe(2+)/Fe(3+) redox pair, which point to oxidative stress as a crucial factor in the development or progress of these neurodegenerative diseases. PMID- 16038517 TI - Antimicrobial susceptibility patterns of contemporary pathogens from uncomplicated urinary tract infections isolated in a multicenter Italian survey: possible impact on guidelines. AB - During 2004 four Italian Laboratories assessed the prevalence of antimicrobial resistance among uropathogens causing acute uncomplicated cystitis in female out patients. A total of 600 urine samples from individuals aged 18-65 were studied. The overall prevalence of Escherichia coli was 85.3%. Klebsiella pneumoniae, Staphylococcus saprophyticus, Proteus mirabilis, Enterococcus faecalis and other rarer species were far less represented. Determination of the antibiotic susceptibility pattern of the entire collection of E. coli (512 organisms) revealed that among the drugs analyzed ampicillin was the least active molecule with only 62.5% of the strains being inhibited. Amoxicillin-clavulanate and cefuroxime displayed a higher potency (87.7% and 89.2% respectively). Cotrimoxazole inhibited only 70.1% of the uropathogens. The three fluoquinolones tested had comparable activity ranging from 83.0% for ciprofloxacin, to 83.6% for levofloxacin and 84.9% for prulifloxacin, indicating an identical spectrum of cross resistance. Nitrofurantoin (96.7%) and fosfomycin (98.6%) were the most potent drugs. Against the whole collection of uropathogens, only cefuroxime, nitrofurantoin and fosfomycin overcame the threshold of 90% activity, with the fluoroquinolones and amoxicillin-clavulanate suffering from about 15% resistance. The results of this survey strongly support the conclusions of recent Italian guidelines concerning the best empiric treatment of UTI in this country today. PMID- 16038518 TI - Pulsed-field gel electrophoresis of human group B streptococci isolated in Brazil. AB - The present study addresses epidemiological aspects of Brazilian human group B streptococci (GBS). GBS (103 isolates) were serotyped with specific rabbit anticapsular antibodies by double diffusion in agarose gels. They represented 3 serotypes: 26 II, 41 III, and 36 V. Thereafter, the strains were characterized by pulsed-field gel electrophoresis (PFGE) of DNA treated with SmaI. DNA restriction band sizes were compared and displayed 54 PFGE profiles that were arranged into 18 patterns. Of the predominant patterns detected for the 41 type III isolates 4 were observed in 15 strains from individuals with infections whereas only 3 were identified in 22 streptococci from healthy carriers. Such differences did not separate types II and V streptococci from carriers and patients. The PFGE method is a sensitive, precise, and powerful tool for discriminating streptococcal strains for epidemiological purposes. PMID- 16038519 TI - Antimicrobial susceptibility, biochemical and genetic profiles of Staphylococcus haemolyticus strains isolated from the bloodstream of patients hospitalized in critical care units. AB - Staphylococcus haemolyticus strains (n=20), responsible of blood stream infections, were consecutively isolated from patients hospitalized in two different wards at high risk of infection. Strains displayed high rate of resistance to oxacillin (90%). All strains but two with decreased susceptibility (MIC = 4 microg/mL), were sensitive to vancomycin. Ten strains were resistant to teicoplanin. Among the strains susceptible to glycopeptides, three displayed heteroresistance to vancomycin and seven to teicoplanin, when tested by Etest technique with 2 x McFarland inoculum. Biochemical reactions allowed to assign strains to eight biotypes, with 11 strains clustering under two main biotype A and biotype B. Pulsed-field-gel-electrophoresis (PFGE) identified 11 different PFGE-types. Seven strains grouping under the major PFGE-type 1 and three strains clustering in PFGE-type 2, closely correlated to biotype A and biotype B respectively. Seven teicoplanin-resistant isolates clustered in the PFGE-type 1, two in the PFGE-type 2 and one in PFGE-type 5. Therefore, teicoplanin-resistant strains were biochemically and genetically related and clonally distributed, despite different clones of S. haemolyticus circulated in the units during the study period. PMID- 16038520 TI - Antimicrobial resistance and characterization of Helicobacter pylori strains isolated from Mexican adults with clinical outcome. AB - Eradication of Helicobacter pylori infection in Mexico is of great importance due to the elevated seroprevalence, however, there is yet very little information about antibiotic resistance rates in H. pylori isolates in our country. We analyzed susceptibility to three antimicrobials used in therapy of 49 H. pylori strains isolated from patients with active chronic gastritis, active chronic gastritis with lymphoid follicles, intestinal metaplasia and gastric cancer. All isolated strains were susceptible to amoxicillin, 28 (58%) were resistant to metronidazole and 2 (4%) were resistant to both clarithromycin and metronidazole. Sequence analysis of the 23S rRNA of the two clarithromycin-resistant strains showed the A2142G mutation in one and A2143G and T2182C mutations in the other. Metronidazole resistance was associated with cagA negative strains with a frequency of 82% (9/11). No significant correlation was found between vacA s/m alleles and metronidazole resistance. PMID- 16038521 TI - Alternative indicators for monitoring the quality of a continuous intervention program on antibiotic prescribing during changing healthcare conditions. AB - We recently published on the impact of a four-phase hospital-wide intervention program designed to optimize the quality of antibiotic use, where a multidisciplinary team (MDT) could modify prescription at the last phase. Because health care quality was changing during the last 5 years (late 1999 to early 2004), we developed certain indicators to monitor the quality of our intervention over time. Different periods were defined as baseline (pre-intervention), initial intervention-active control, pre-crisis control, crisis control, post-crisis control and end of crisis control. Major indicators were rates of prescription modification by the MDT; prescription for an uncertain infection and a novel index formula (RIcarb) to estimate the rationale for carbapenem use. We assessed 2115 antimicrobial prescriptions. Modification of prescription rate was 30% at the beginning and decreased thereafter up to stable levels. Rate of prescriptions ordered for cases of both uncertain infection and unknown source of infection decreased significantly after intervention (i.e. from baseline to active control). In contrast, a doubling of culture-directed prescriptions was observed between these periods. RIcarb values lower and higher than 60% (modal, cut-off) were assumed as carbapenem overuse and underuse, respectively. Overuse was observed at the pre-intervention, while pronounced underuse was shown during the crisis (RIcarb, 45% and 87%, respectively). The present study demonstrates that certain indicators, other than the widely adopted impact outcomes, are a suitable tool for monitoring the quality of a continuous, long-term, active intervention on antimicrobial prescribing practice, especially when applied in a changing healthcare setting. PMID- 16038523 TI - Antagonism of HSV-tk transfection and ganciclovir treatment on chemotherapeutic drug sensitivity. AB - Our study focused on the influence of herpes simplex virus thymidine kinase (HSV tk) expression and ganciclovir (GCV) treatment on the sensitivity of C6 glioma cells to frequently used chemotherapeutic drugs, i.e. adriamycin (ADR), cisplatin (CDDP), 5-fluorouracil (5-FU), and methotrexate (MTX). Transfection with HSV-tk revealed an increased sensitivity to GCV and CDDP and a decreased sensitivity to ADR and MTX. No significant differences were found in sensitivity to 5-FU. Combined treatment in a HSV-tk negative cell line revealed an additive effect when GCV was combined with ADR, whereas an antagonistic effect was found when GCV was combined with CDDP, 5-FU, or MTX. Comparable results were obtained in an HSV tk positive cell line, apart from CDDP, which showed an additive effect. In conclusion, both HSV-tk transfection and subsequent GCV treatment can influence the sensitivity of tumor cells to various chemotherapeutic drugs in an antagonistic manner. Therefore, combining HSV-tk/GCV gene therapy with chemotherapy might not always be beneficial. PMID- 16038522 TI - Treatment of childhood complicated community-acquired pneumonia with amoxicillin/sulbactam. AB - The clinical and microbiological efficacy, as well as the tolerability of the amoxicillin/sulbactam combination as empiric treatment for complicated community acquired pneumonia (CCAP) were evaluated in children from 3 months to 15 years with CCAP who were randomized 1:1 to receive either amoxicillin/sulbactam or cefuroxime. Of 234 patients hospitalized with CCAP in the study period (June, 1999-April, 2002), 62 patients qualified for the study: 32 received amoxicillin/sulbactam and 30 cefuroxime. Two were excluded. Demographic and clinical data showed that both groups were comparable at entry. One etiologic agent was identified in 55% of the patients, with Streptococcus pneumoniae being the most frequent. After treatment, the days of fever, duration of intravenous treatment, and hospitalization stay were similar in both groups. Overall favorable clinical responses were comparable: 97% for amoxicillin/sulbactam vs 100% for the comparative therapy. There was good tolerance to both drugs. Amoxicillin/sulbactam produced a satisfactory therapeutic outcome similar to that of cefuroxime for treatment of CCAP, and may be an appropriate choice for the treatment of this serious pediatric infection. PMID- 16038524 TI - High dose chemotherapy and transplantation of hematopoietic progenitors from murine D3 embryonic stem cells. AB - Differentiating embryonic stem (ES) cells are an increasingly important source of hematopoietic progenitors, useful for both basic research and clinical applications. To date, characteristics of specific factors capable of influencing hematopoietic cell fate from ES cells remains elusive. We report that mMSC Feeder Layer and the combination of VEGF, SCF and TPO strongly promote hematopoietic differentiation. The results showed that the cells induced from ES-D3 expressed hematopoietic progenitor antigens (CD34 and CD117), myelocyte cell antigen (CD11b), erythrocyte cell antigen (Ter119), and transcription factors (Flk-1, GATA-2, SCL, beta-H1 and beta-major). Furthermore, those induced differentiated cells were injected into female C57BL/6 mice which were treated with high dose topotecan chemotherapy to restore part of their blood system function. We observed rapid white blood cell recovery, which suggested that the infusion of differentiated cells has a positive impact on hematopoiesis. The Sry gene in peripheral blood, bone marrow and spleen of transplanted female mice was confirmed by PCR analysis, which affirmed the existence of the chimera. PMID- 16038525 TI - In vitro and in vivo reversal of cancer cell multidrug resistance by 2',4' dihydroxy-6'-methoxy-3',5'-dimethylchalcone. AB - 2',4'-Dihydroxy-6'-methoxy-3',5'-dimethylchalcone (DMC) isolated from the buds of Cleistocalyx operculatus, was investigated for its reversal effects on cancer cell multidrug resistance. DMC potentiated the cytotoxicity of the chemotherapeutic agent doxorubicin to drug-resistant KB-A1 cells. When 5 microM DMC was present simultaneously with doxorubicin, the IC50 of DOX on KB-A1 cells decreased from 13.9 +/- 0.7 microg/ml to 3.6 +/- 0.7 microg/ml. A human carcinoma xenograft model was established with the KB-A1 cell line. DMC could sensitize the tumors to doxorubicin as indicated by a considerable reduction in tumor weight. DMC increased the intracellular accumulation of doxorubicin in KB-A1 cells. When KB-A1 cells were exposed to 10 microg/ml doxorubicin combined with 5, 10, 20 microM DMC for 4 hours, the intracellular concentrations of doxorubicin were increased 1.4-, 1.8-, 3.1-fold, respectively, in comparison with doxorubicin alone treatment. All results indicated that DMC had reversal effects on the multidrug resistance phenotype. PMID- 16038526 TI - Pharmacokinetics of bolus 5-fluorouracil: relationship between dose, plasma concentrations, area-under-the-curve and toxicity. AB - The pharmacokinetics of 5-fluorouracil (5FU) have been related to toxicity and antitumor activity, in particular for continuous infusion schedules, but to a lesser extent for frequently used bolus injections. The use of intensive sampling schedules limits the application of pharmacokinetics to optimize individual dosing or to define the ideal combination with other drugs. We therefore reanalyzed a pharmacokinetic study in order to develop a limited sampling schedule. Patients received escalating doses of 5FU at 500, 600 and 720 mg/m2 as a bolus until toxicity developed. Blood samples were analyzed until 24 h after administration. The area under the concentration time curve from 0-90 min (AUC(0 90)) was strongly correlated with dose and also with toxicity (p = 0.0009). The 5FU concentrations at 30 and 60 min were correlated to the AUC(30-240) and to that of the AUC(0-90) (r2 = 0.970). The use of limited sampling (30, 60, 90 min) in a patient given 353 mg/m2 5FU with severe toxicity at initial dosing at 500 mg/m2 revealed that the AUC(0-90) at 353 mg/m2 was higher than the normal AUC(0 90) for 500 mg/m2. This patient appeared to have an 8-fold lower activity of the 5FU degradation enzyme dihydropyrimidine dehydrogenase. Limited sampling will allow us to define potential aberrant kinetics of pharmacokinetic interaction of 5FU with other drugs being developed for treatment of colorectal cancer. PMID- 16038527 TI - Oligodendrogliomas lacking O6-methylguanine-DNA-methyltransferase expression. AB - O6-Methylguanine-DNA-Methyltransferase (MGMT) is a DNA repair protein considered to be a chemosensitivity predictor. We evaluated the immunohistochemical MGMT expression in 28 consecutive oligodendroglial tumors (21 oligodendrogliomas, 5 mixed oligoastrocytomas, and 2 glioblastomas with prominent oligodendroglial features; 13 treated with CCNU) and compared it with that of 13 glioblastomas. Twenty-six (93%) oligodendroglial tumors were MGMT-negative, 2 (7%) were MGMT positive. Twelve (92%) patients treated with CCNU had MGMT-negative lesions and their median survival was 73 months; 1 patient had an MGMT-positive oligodendroglioma and is alive at 28 months. Three (23%) glioblastomas were MGMT negative and 10 (77%) MGMT-positive. The lower MGMT expression in oligodendroglial tumors compared to glioblastomas (P < 0.05), which have different chemosensitivity, suggests a possible role of MGMT in the determination of chemoresistance. Nevertheless, the heterogeneous outcome of our MGMT-negative oligodendroglial tumors treated with CCNU, indicates that MGMT expression alone is insufficient to predict the response to alkylating drugs, presumably because of the numerous mechanisms involved. PMID- 16038528 TI - Effect of sedation with midazolam or propofol on patient's comfort during cancer chemotherapy infusion: a prospective, randomized, double-blind study in breast cancer patients. AB - Nausea, vomiting and anxiety related to administration of cancer chemotherapy remain significant problems. This randomized, clinical trial was undertaken to evaluate the effect of sedation on the patient's comfort during chemotherapy infusion in patients with breast cancer. Forty-five breast cancer patients were randomized into three groups: Group I--chemotherapy, control, Group II- midazolam+chemotherapy, and Group III--propofol+chemotherapy. Nausea occured in 87% and vomiting in 13% of the patients in the control group, while none of the sedated subjects had these side-effects, although 76% of them had experienced then during previous cycles of chemotherapy. Compared with the control group, post-chemotherapy anxiety scores also improved with the addition of midazolam or propofol. Eighty percent of the subjects declared that they would prefer the sedative-containing regimen for their further cycles. Sedation with midazolam or propofol may improve the patient's comfort, and provide better control of chemotherapy-related side effects during chemotherapy infusion in breast cancer patients. PMID- 16038529 TI - Oncologic emergencies secondary to advanced colorectal cancer successfully treated with oxaliplatin/5-fluorouracil/leucovorin: report of three cases. AB - Metastatic/advanced colorectal cancer is considered a resistant disease and oncologic emergencies secondary to advanced disease may be regarded with a nihilistic attitude. The objective of this report is to emphasize the efficacy of the oxaliplatin/5-fluorouracil/leucovorin regimen (FOLFOX-4) in three patients presenting oncologic emergencies secondary to advanced colon cancer. The first case was a 40-year-old man with severe respiratory insufficiency due to massive carcinomatous lymphangitis; subsequently a cecal adenocarcinoma was diagnosed. The patient's conditions became life-threatening and he was admitted to the intensive care unit. The second case was a 41-year-old woman presenting with fever, abdominal mass and pain. Ultrasound and CT-scan revealed two hepatic masses (13 x 15 and 15 x 20 cm), diagnosed as liver metastases from colon cancer. The patient's condition deteriorated with intestinal obstruction secondary to the large left liver mass. The third case was a 58-year-old woman presenting with hepatic mass, fever and weight loss. Ultrasound and CT-scan showed a liver lesion occupying the right lobe (12 x 14 cm). Ultrasonically-guided biopsy and colonoscopy showed liver metastases from cecal cancer. A 5 fluorouracil/leucovorin regimen failed to improve her clinical condition and she had disease progression, inferior vena cava neoplastic thrombosis and right hydronephrosis. All three patients rapidly improved after a few cycles of oxaliplatin-containing chemotherapy. These cases demonstrate that even patients with advanced colorectal cancer presenting with oncologic emergencies and life threatening conditions can be successfully treated with the FOLFOX-4 regimen. PMID- 16038530 TI - Adverse interaction between capecitabine and warfarin resulting in altered coagulation parameters and bleeding: case report and review of the literature. AB - Capecitabine can interact with warfarin, resulting in altered coagulation parameters and bleeding. Four cases have been reported. We describe a fifth case with life-threatening interaction between these two drugs. A 67-year-old female with metastatic breast cancer developed hemorrhagic blisters, purpura and ecchymoses. She had been well controlled on long-term warfarin (5 mg/day). Capecitabine was initiated 4.5 weeks prior to the bleeding episode. Laboratory work-up revealed an international normalized ratio of 8.56, partial prothrombin time of 61 seconds and prothrombin time of 5.2%. The coagulation parameters gradually normalized within 4 days following vitamin K administration and discontinuation of capecitabine and warfarin. Careful monitoring of coagulation parameters and proper adjustment of the warfarin dose are required in patients taking warfarin and capecitabine concomitantly. PMID- 16038531 TI - Severe acute enteritis in a multiple myeloma patient receiving bortezomib and spinal radiotherapy: case report. AB - Proteasome inhibitors have been reported to enhance radiosensitivity in vitro. A case of potential clinical interaction between bortezomib, a proteasome inhibitor, and spine radiation is reported. A woman undergoing palliative radiotherapy to the T12 -S2 spine with concurrent bortezomib developed unexpectedly severe, acute radiation enteritis requiring hospital admission. Clinicians are advised to consider the potential for interactions of bortezomib with radiotherapy when the two agents are used simultaneously in the clinic. PMID- 16038532 TI - Azole resistance in neonatal intensive care units in Argentina. AB - The aim of this study was to determine the antifungal susceptibility profile and to detect resistant strains of yeast species isolated from neonates in Intensive Care Units. 92 strains isolated from 25 bloodstream cultures, 20 venous catheters, 23 suprapubic aspirations and 24 rectal swabs were studied. A Candida glabrata strain resistant to fluconazole was detected. Candida krusei appeared with its inherent resistance to fluconazole and showed cross-resistance to itraconazole. Two Candida albicans strains were resistant to azoles, one to itraconazole and the other to fluconazole with a high minimum inhibitory concentration (MIC) for itraconazole. All Candida tropicalis strains were susceptible to fluconazole but two of them showed resistance to itraconazole. The detection of resistant strains in neonates whom had not received previous antifungal therapy is noteworthy. The variations in the epidemiology of fungal infections observed and the antifungal resistance detected emphasize the importance of performing a regular surveillance to observe and to assess them. PMID- 16038533 TI - The transgenic Arabidopsis plant system, pER8-GFP, as a powerful tool in searching for natural product estrogen-agonists/antagonists. AB - The transgenic Arabidopsis plant system, pER8-GFP, may be used as a powerful tool in searching for natural estrogen-agonists/antagonists. Among selected plant extracts and natural products, the method was able to distinguish active extracts (e.g., Glycine max and Pueraria lobata) and pure compounds (e.g., 17beta estradiol (1), genistein (10), and daidzein (11)) and also to distinguish effects of structural changes on activity. Thus, this rapid sensitive system was proven to be suitable for the discovery of natural products with estrogenic activity. PMID- 16038534 TI - Antineoplastic agents. 545. Isolation and structure of turbostatins 1-4 from the Asian marine mollusk Turbo stenogyrus. AB - The cancer cell line bioassay-guided separation of an extract from the marine mollusk Turbo stenogyrus led to the isolation of four new cerebrosides designated turbostatins 1-4 (1-4). The structure of each glycolipid was determined by interpreting results of a series of HR-APCI-MS and NMR (1D and 2D) spectral analyses. All four turbostatins exhibited significant (GI50 0.15-2.6 microg/mL) cancer cell growth inhibition against the murine P388 lymphocytic leukemia and a panel of human cancer cell lines. PMID- 16038535 TI - Acylphloroglucinol derivatives from Mahurea palustris. AB - Five new acylphloroglucinol derivatives, mahureones A-E (1, 3-6), have been isolated from the leaves of Mahurea palustris, and their structures determined by spectroscopic means. During the isolation process, several byproducts (7-9) were formed by reaction of one of the isoprenyl side chains with TFA, water, and acetonitrile. All the compounds were assayed for their ability to inhibit human DNA polymerase beta. The most active compounds, mahureones A (1) and D (5), exhibited IC50 values in the 10 microM range. PMID- 16038536 TI - Expanding the ChemGPS chemical space with natural products. AB - Recently various attempts have been made to increase the efficacy and precision of chemical libraries used in high-throughput screening (HTS) drug discovery approaches. One such approach is ChemGPS, which provides a defined chemical space for prescreening evaluation of chemical compound properties or virtual dereplication. In the present study, ChemGPS has been applied to a set of natural products shown to exhibit cyclooxygenase-1 and/or -2 (COX-1/2) inhibition. With the purpose of defining chemical properties and linking these to the observed mode of enzyme inhibition, this resulted in two lines of reasoning. On one hand several specific features of these compounds have been identified and discussed. Overall COX inhibition was frequently correlated with the presence of at least one ring in the structure, fragments exhibiting structural rigidity, and a relatively large molecular size. The concept "size" includes several parameters, e.g., molecular volume, weight, and number of bonds. On the other hand, and possibly even more important, was the unexpected finding that the natural products studied to a large extent fell outside the defined ChemGPS chemical space. Therefore, we also propose an expanded space for natural products: ChemGPS NP. PMID- 16038537 TI - Structural analyses of metabolites of phenolic 1-benzyltetrahydroisoquinolines in plant cell cultures by LC/NMR, LC/MS, and LC/CD. AB - The metabolism of the phenolic 1-benzyltetrahydroisoquinoline alkaloids was studied in cell cultures of Macleaya and Corydalis species. The crude alkaloid fraction obtained from feeding experiments was investigated by application of the combined LC/NMR and LC/APCI-MS (/MS) techniques. Several metabolites were detected, and their structures (6 and 8-14) were identified. Bioconversion of the phenolic 1-benzyltetrahydroisoquinoline (2) into the pseudoprotoberberine (8) was demonstrated for the first time. LC/APCI-MS and LC/CD experiments carried out on a chiral column permitted the deduction of the major enantiomeric form of the chiral metabolites. Thus, the combination of NMR, MS, and CD data permitted the structural elucidation and stereochemical analysis of the metabolites in the extract matrix solution, without isolation and sample purification prior to the coupling experiments. PMID- 16038539 TI - Bioactive constituents of the leaves of Phyllanthus polyphyllus var. siamensis. AB - A dichloromethane extract of the leaves of Phyllanthus polyphyllus var. siamensis afforded two new monoacetylated triterpene arabinosides (1 and 2) and 15 known compounds. The structures of the new compounds were elucidated on the basis of spectroscopic data interpretation. The isolates and some chemical transformation products were evaluated biologically, and the presence of an arabinosyl moiety among the hederagenin glycosides evaluated was found to be important for the modulation of cytotoxic activity against cancer cells. PMID- 16038538 TI - Acetylenic strongylodiols from a Petrosia (Strongylophora) Okinawan marine sponge. AB - Seven new long-chain acetylenic alcohols, strongylodiols D-J, were isolated from an Okinawan marine sponge of the genus Petrosia (Strongylophora). The structures of these compounds were elucidated on the basis of the results of spectroscopic analysis and chemical reaction. Analysis of the MNA esters of the acetylenic alcohols disclosed that these compounds were each an enantiomeric mixture in a different ratio. PMID- 16038540 TI - Xanthone and sesquiterpene derivatives from the fruits of Garcinia scortechinii. AB - The fruits of Garcinia scortechinii afforded 10 new compounds: four caged tetraprenylated xanthones (scortechinones Q-T, 1-4), four rearranged xanthones (scortechinones U-X, 5-8), and two sesquiterpene derivatives (scortechterpenes A, B, 9, 10), together with 14 known compounds: one sesquiterpene, two biflavonoids, and 11 caged-polyprenylated xanthones. Their structures were elucidated by analysis of spectroscopic data and comparison of the NMR data with those reported previously. All xanthone derivatives were evaluated for antibacterial activity against methicillin-resistant Staphylococcus aureus. PMID- 16038541 TI - Lupane triterpenoids from Maytenus species. AB - Five new lupane triterpenes (1-5), in addition to 24 known ones, were isolated from Maytenus cuzcoina and M. chiapensis. Their structures were elucidated on the basis of spectroscopic analysis, including homonuclear and heteronuclear correlation NMR (COSY, ROESY, HSQC, and HMBC) experiments. The compounds were assayed for antimicrobial and cytotoxic activities, with 3-epi-betulinic acid and 28,30-dihydroxy-3-oxolup-20(29)-ene showing moderate cytotoxicity. PMID- 16038542 TI - Neurotoxic meroditerpenoids from the tropical marine brown alga Stypopodium flabelliforme. AB - Brine shrimp toxicity and TLC analysis guided the isolation of five new and biologically active meroditerpenoids [2beta,3alpha-epitaondiol (1), flabellinol (2), flabellinone (3), stypotriolaldehyde (4), and stypohydroperoxide (5)] along with five known compounds from the marine brown alga Stypopodium flabelliforme collected in Papua New Guinea. The planar structures of compounds 1-5 were determined by extensive spectroscopic analysis (1D and 2D NMR, LRMS, HRMS, IR, and UV), while relative configuration was determined by 1D and 2D NOE experiments. X-ray crystallography confirmed the relative configuration of 2beta,3alpha-epitaondiol (1), and the modified Mosher's ester method was used to establish its absolute configuration. All of the new metabolites were moderately toxic to murine neuro-2a cells (LC50 2-25 microM), and three [2beta,3alpha epitaondiol (1), flabellinol (2), and flabellinone (3)] possessed potent sodium channel blocking activity. Stypotriolaldehyde (4) had a biphasic effect on the concentration of intracellular Ca2+ in rat cerebellar granule neurons (CGN). The previously known compound, stypoldione (6), also modulated intracellular calcium concentration and was cytotoxic in CGN. Metabolites 2beta,3alpha-epitaondiol (1), flabellinol (2), and flabellinone (3) displayed moderate cytotoxicity to the NCI H460 human lung cancer cell line. PMID- 16038543 TI - Myo-inositol-derived glycolipids with anti-inflammatory activity from Solanum lanceolatum. AB - Lanceolitols A1-A7 (1-7) and B1-B7 (9-15), two series of new myo-inositol-derived glycolipid analogues, in which a sugar moiety is replaced by a fatty acid esterified myo-inositol moiety, were isolated from the leaves of Solanum lanceolatum. Their structures were elucidated on the basis of spectroscopic analysis (1H NMR, 13C NMR, 1H-1H COSY, HMQC, HMBC, and HRFABMS), as well as chemical analysis. All the compounds showed in vivo anti-inflammatory activity against ear edema in mice produced by 12-O-tetradecanoylphorbol-13-acetate (TPA). In vitro enzyme inhibition studies showed that the mixture of lanceolitols A1-A7 inhibited by 58.56% phospholipase A2 from bee venom, while the mixture of lanceolitols B1-B7 was cyclooxygenase-2 (COX-2) inhibitors (IC50 = 237 microM). PMID- 16038544 TI - Isolation of an antiviral polysaccharide, nostoflan, from a terrestrial cyanobacterium, Nostoc flagelliforme. AB - A novel acidic polysaccharide, nostoflan, was isolated from a terrestrial cyanobacterium, Nostoc flagelliforme. Nostoflan exhibited a potent anti-herpes simplex virus type 1 (HSV-1) activity with a selectivity index (50% cytotoxic concentration/50% inhibitory concentration against viral replication) of 13,000. Sugar composition and methylation analyses revealed that it was mainly composed of -->4)-D-Glcp-(1-->, -->6,4)-D-Glcp-(1-->, -->4)-D-Galp-(1-->, -->4)-D-Xylp-(1- >, D-GlcAp-(1-->, D-Manp-(1--> with a ratio of ca. 1:1:1:1:0.8:0.2. Two pyridylaminated oligosaccharides were prepared by partial acid hydrolysis and pyridylamination. On the basis of MALDI-TOF-MS and NMR analyses, they were found to be beta-D-Glcp-(1-->4)-D-Xyl-PA and beta-D-GlcAp-(1-->6)-beta-D-Glcp-(1-->4)-D Gal-PA. From these results, nostoflan might be mainly composed of the following two types of sugar sequence: -->4)-beta-D-Glcp-(1-->4)-D-Xylp-(1--> and -->4) [beta-D-GlcAp-(1-->6)-]-beta-D-Glcp-(1-->4)-D-Galp-(1-->. Besides anti-HSV-1 activity, nostoflan showed potent antiviral activities against HSV-2, human cytomegalovirus, and influenza A virus, but no activity against adenovirus and coxsackie virus was observed. Therefore, nostoflan has a broad antiviral spectrum against enveloped viruses whose cellular receptors are carbohydrates. Furthermore, nostoflan showed no antithrombin activity, unlike sulfated polysaccharides. PMID- 16038545 TI - Cytotoxic activity of some natural and synthetic guaianolides. AB - Several natural guaianolides and synthetic derivatives of repin (1) were tested and found to be active against tumor cell replication. Repin (1) and both mono- and di-halohydrin analogues (2, 7-9, 11, 12) showed significant antitumor potency. A more effective compound (17) was obtained by esterificating repin with the paclitaxel side chain. PMID- 16038546 TI - Azadirachtin derivatives from seed kernels of Azadirachta excelsa. AB - Three new azadirachtin derivatives, named azadirachtins O-Q (1-3), along with the known azadirachtin B (4), azadirachtin L (5), azadirachtin M (6) 11alpha azadirachtin H (7), 11beta-azadirachtin H (8), and azadirachtol (9) were isolated from seed kernels of Azadirachta excelsa. Their structures were established by spectroscopic techniques, and the structure of 3 was confirmed by X-ray analysis. Compounds 1-7 and 9 exhibited toxicity to the diamondback moth (Plutella xylostella) with an LD50 of 0.75-1.92 microg/g body weight, in 92 h. PMID- 16038547 TI - Eunicellin-based diterpenoids, australins A-D, isolated from the soft coral Cladiella australis. AB - The EtOAc extract of a Taiwanese soft coral, Caldiella australis, yielded four new eunicellin-based diterpenoids, australins A-D (1-4). The chemical structures of these metabolites were determined on the basis of extensive spectroscopic (including 1D and 2D NMR) analyses and by comparison of their NMR spectral data with those of related metabolites. Metabolite 2 was found to possess a moderate cytotoxic activity against selected breast and liver cancer cell lines. PMID- 16038548 TI - New bisbenzylisoquinolines, fatty acid amidic aporphines, and a protoberberine from Formosan Cocculus orbiculatus. AB - Two new bisbenzylisoquinoline alkaloids, (+)-coccuorbiculatine A (2) and (+)-10 hydroxyisotrilobine (3), two new amidic aporphines, a mixture of (+) laurelliptinhexadecan-1-one (6) and (+)-laurelliptinoctadecan-1-one (7), and one new protoberberine (-)-4-methoxy-13,14-dihydrooxypalmatine (8) have been isolated from the stems of Taiwanese Cocculus orbiculatus. The structures were established on the basis of extensive analysis of spectroscopic data and by comparison with known related metabolites. Cytotoxicity of the isolated compounds was examined toward HepG2, Hep3B, MCF-7, and MDA-MB-231 cancer cell lines. Alkaloids 1 and (-) sinococuline (9) showed significant inhibitory activity against the target cell lines. PMID- 16038549 TI - Nocardimicins A, B, C, D, E, and F, siderophores with muscarinic M3 receptor inhibiting activity from Nocardia sp. TP-A0674. AB - In the screening for muscarinic M3 receptor binding inhibitors from microbial secondary metabolites, the extract of Nocardia sp. TP-A0674 was found to be highly active. Bioassay-guided fractionation of it led to the isolation of six new siderophores, nocardimicins A (1), B (2), C (3), D (4), E (5), and F (6), as active principles. Their chemical structures were determined by spectroscopic and degradation analysis. Of these congeners, nocardimicin B (2) inhibited the binding of tritium-labeled N-methylscopolamine to the muscarinic M3 receptor most potently with a Ki value of 0.13 microM. Compound 2 showed more selective activity to M3 and M4 receptors than other subtypes. PMID- 16038550 TI - Nitrogen-containing compounds from Salvia miltiorrhiza. AB - Five new N-containing compounds, neosalvianen (1), salvianen (2), salvianan (3), salviadione (4), and 5-(methoxymethyl)-1H-pyrrole-2-carbaldehyde (5), were isolated from Salvia miltiorrhiza. Their structures were mainly established by spectroscopic methods. Neosalvianen (1) and its analogues (6a, 6b) were synthesized for spectroscopic data comparison. Compounds 1, 2, 4, and 6a were evaluated for their cytotoxic activities against selected cancer cell lines. Among these components, salvianen (2) exhibited the most potent cytotoxicity with a CD50 range of 30.4-39.5 microM against HeLa (cervical epitheloid carcinoma), HepG2 (hepatocellular carcinoma), and OVCAR-3 (ovarian adenocarcinoma) cell lines in a dose-dependent manner. The cytotoxicities of the tested compounds were not specific and showed similar activities to the selected cancer cell lines. PMID- 16038551 TI - Phenanthroindolizidine alkaloids from the stems of Ficus septica. AB - In addition to six known phenanthroindolizidine alkaloids, eight new alkaloids, namely, ficuseptines B-D (1-3), 10R,13aR-tylophorine N-oxide (4), 10R,13aR tylocrebrine N-oxide (5), 10S,13aR-tylocrebrine N-oxide (6), 10S,13aR isotylocrebrine N-oxide (7), and 10S,13aS-isotylocrebrine N-oxide (8), were isolated from a methanol extract of the stems of Ficus septica. The structures of the new compounds were elucidated by means of spectroscopic data interpretation. Cytotoxicity of some of these alkaloids was assessed in vitro using the HONE-1 and NUGC cell lines. PMID- 16038552 TI - C20-diterpenoid alkaloids from Delphinium trifoliolatum. AB - Three new C20-diterpenoid alkaloids, trifoliolasines D-F (1-3), were isolated from the aerial parts of Delphinium trifoliolatum, and their structures were determined by the interpretation of spectroscopic data and by the single-crystal X-ray crystallographic analysis of 1. PMID- 16038553 TI - Latifolians A and B, novel JNK3 kinase inhibitors from the Papua New Guinean plant Gnetum latifolium. AB - As part of our search for natural products active against the JNK3 kinase, two novel, charged benzylisoquinolines, latifolian A (1) and latifolian B (2), were isolated from the stem bark of the Papua New Guinean vine Gnetum latifolium. The planar structures were determined through detailed 2D NMR analysis. The relative configurations were assigned after examination of the ROESY data and through detailed molecular modeling studies. PMID- 16038554 TI - Koniamborine, the first pyrano[3,2-b]indole alkaloid and other secondary metabolites from Boronella koniambiensis. AB - Two new alkaloids, (-)-cis-1,2-dihydroxy-1,2-dihydromedicosmine (3) and koniamborine (4), have been isolated from Boronella koniambiensis aerial parts. Their structures have been established from NMR and mass data. Koniamborine is a novel type of alkaloid, which derives from the pyrano[3,2-b]indole basic skeleton, described for the first time from nature. 6-Methoxy-1-methylisatin, also present in the plant material, can be considered biogenetically as a degradation product of the fused pyrone ring of 4. PMID- 16038555 TI - Cytotoxic diterpenoids from the soft coral Sinularia microclavata. AB - The soft coral Sinularia microclavata collected from the bay of Sanya, Hainan Island, China, yielded a new diterpenoid, microclavatin (1), and a known cembranolide, capillolide (2). The structure of compound 1 was determined on the basis of spectroscopic methods and X-ray single-crystal diffraction analysis. Microclavatin (1) showed cytotoxic activities against tumor cell lines KB and MCF with IC50 values of 5.0 and 20.0 microg/mL, respectively, and capillolide (2) showed potent cytotoxic activity against tumor cell lines (A-549) with an IC50 value of 0.5 microg/mL. PMID- 16038556 TI - A labdane diterpene glucoside from the rhizomes of Curcuma mangga. AB - A new labdane diterpene glucoside, curcumanggoside (1), together with nine known compounds, including labda-8(17),12-diene-15,16-dial (2), calcaratarin A (3), zerumin B (4), scopoletin, demethoxycurcumin, bisdemethoxycurcumin, 1,7-bis(4 hydroxyphenyl)-1,4,6-heptatrien-3-one, curcumin, and p-hydroxycinnamic acid, have been isolated from the rhizomes of Curcuma mangga. Their structures were determined using a combination of 1D (1H NMR, 13C NMR, DEPT) and 2D (COSY, HSQC, HMBC) NMR techniques. All diarylheptanoids and scopoletin showed significant antioxidant activity. Zerumin B, demethoxycurcumin, bisdemethoxycurcumin, and curcumin also exhibited cytotoxic activity against a panel of five human tumor cell lines. PMID- 16038557 TI - Flavonoids from the stem of Eriophorum scheuchzeri. AB - Activity-guided isolation of antifungal, antibacterial, and antioxidant compounds from the dichloromethane extract of the aerial parts of Eriophorum scheuchzeri led to the isolation of eight flavonoids. Three isoflavones and one flavanone are new natural compounds. The isolation of the bioactive compounds was performed by centrifugal partition chromatography. Fractions and pure compounds were monitored by a direct TLC bioautographic assay. Their structures were elucidated by spectroscopic and chemical methods. PMID- 16038558 TI - Dihydrostilbene derivatives from Glycyrrhiza glabra leaves. AB - Four new dihydrostilbenes, alpha,alpha'-dihydro-3,5-dihydroxy-4'-acetoxy-5' isopentenylstilbene (1), alpha,alpha'-dihydro-3,3',4'-trihydroxy-5-O-isopentenyl 6-isopentenylstilbene (2), alpha,alpha'-dihydro-3,5,3'-trihydroxy-4' methoxystilbene (3), and alpha,alpha'-dihydro-3,3'-dihydroxy-5beta-d-O glucopyranosyloxy-4'-methoxystilbene (4), together with seven known flavonoids, glabranin isomer, naringenin, lupiwighteone, pinocembrin 7-O-glucoside, astragalin, isoquercitrin, vicenin II, and the inositol, pinitol, were isolated from the leaves of Glycyrrhiza glabra grown in Sicily. The structures of 1-4 were elucidated by spectroscopic methods. PMID- 16038559 TI - 3-Nitropropionic acid (3-NPA), a potent antimycobacterial agent from endophytic fungi: is 3-NPA in some plants produced by endophytes? AB - 3-Nitropropionic acid (3-NPA, 1) was found in extracts of several strains of endophytic fungi. 3-NPA (1) exhibited potent antimycobacterial activity with the minimum inhibition concentration of 3.3 microM, but was inactive against NCI H187, BC, KB, and Vero cell lines. Endophytes were found to produce high levels of 3-NPA (1), and therefore 3-NPA (1) accumulated in certain plants may be produced by the associated endophytes. 3-NPA (1) may be used as a chemotaxonomic marker for endophytic fungi. The structure of 3-hydroxypropionic acid, a nematicidal agent, should be revised to 3-NPA (1). PMID- 16038560 TI - Structural revision of aspernigrin A, reisolated from Cladosporium herbarum IFB E002. AB - Aspernigrin A was reisolated as a secondary metabolite of the Cynodon dactylon associated endophytic fungus Cladosporium herbarum IFB-E002 coproducing rubrofusarin B, fonsecinone A, 7-hydroxy-4-methoxy-5-methylcoumarin, orlandin, kotanin, and 3beta,5alpha,6beta-trihydroxyergosta-7,22-diene. The structure of aspernigrin A, previously elucidated to be 4-benzyl-6-oxo-1,6-dihydropyridine-3 carboxamide (1), was revised as 6-benzyl-4-oxo-1,4-dihydropyridine-3-carboxamide (2) on the basis of its additional NMR spectroscopic data and the X-ray crystallographic analysis. PMID- 16038561 TI - Gesashidine A, a beta-carboline alkaloid with an imidazole ring from a Thorectidae sponge. AB - A new beta-carboline alkaloid with an imidazole ring, gesashidine A (1), has been isolated from an Okinawan marine sponge of the family Thorectidae (SS-1035), and the structure was elucidated from the spectroscopic data. PMID- 16038562 TI - Steroids from the Antarctic octocoral Anthomastus bathyproctus. AB - Seven new steroids, compounds 1-7, were isolated from the Antarctic octocoral Anthomastus bathyproctus. The structures of the new metabolites have been established by analysis of their spectroscopic data. The in vitro cytotoxicity has been tested against three human tumor cell lines. PMID- 16038563 TI - Polyhydroxylated spirostanol saponins from the tubers of Dioscorea polygonoides. AB - Three new polyhydroxylated spirostanol saponins (1-3) were isolated from the tubers of Dioscorea polygonoides. The structures of these new compounds were determined on the basis of extensive spectroscopic analysis and the results of acid or enzymatic hydrolysis as (23S,24R,25S)-23,24-dihydroxyspirost-5-en-3beta yl O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranoside (1), (23S,25R) 12alpha,17alpha,23-trihydroxyspirost-5-en-3beta-yl O-alpha-L-rhamnopyranosyl-(1- >2)-beta-D-glucopyranoside (2), and (23S,25R)-14alpha,17alpha,23 trihydroxyspirost-5-en-3beta-yl O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D glucopyranoside (3), respectively. PMID- 16038564 TI - Gaudichanolides A and B, clerodane diterpenes from Baccharis gaudichaudiana. AB - Two new clerodane diterpenoids, gaudichanolides A (1) and B (2), were isolated from the dried twigs of Baccharis gaudichaudiana, together with 7-oxo-16,19 dihydroxy-3,4-dehydroclerodan-15,20-diacid dilactone, spathulenol, 4,10 aromadendranediol, kobusone, trans-cosanyl ferulate, and defuscin. The structures and relative stereochemistry of 1 and 2 were elucidated by detailed 2D NMR spectroscopic experiments, and the absolute stereochemistry of 1 was determined by X-ray crystallographic analysis. PMID- 16038565 TI - Constituents of the pericarp of Garcinia subelliptica. AB - A new benzophenone, garcinielliptone FA (1), and a new benzoylphloroglucinol, garcinielliptone FB (2), along with six known compounds, were isolated from the pericarp of Garcinia subelliptica. The structures and relative configurations of 1 and 2 were elucidated by spectroscopic methods and supported by computer generated molecular modeling. Compound 2 exhibited cytotoxic activity against several human cancer cell lines. PMID- 16038566 TI - Morphinane alkaloids with cell protective effects from Sinomenium acutum. AB - One new morphinane alkaloid, sinomenine N-oxide (1), and one new natural occurring morphinane alkaloid, N-demethylsinomenine (2), together with six known alkaloids, 7,8-didehydro-4-hydroxy-3,7-dimethoxymorphinan-6-ol (3), sinomenine (4), sinoacutine (5), N-norsinoacutine, acutumine, and acutumidine, were isolated from the stems of Sinomenium acutum. Their structures were elucidated on the basis of spectroscopic analysis and chemical methods. Compounds 2, 3, and 5 have protective effects against hydrogen peroxide-induced cell injury. PMID- 16038567 TI - Terpenoids from the roots and rhizomes of Nardostachys chinensis. AB - A new diterpene, 10-isopropyl-2,2,6-trimethyl-2,3,4,5-tetrahydronaphtha[1,8 bc]oxocine-5,11-diol (1), and a new monoterpene, 6-hydroxy-7-(hydroxymethyl)-4 methylenehexahydrocyclopenta[c]pyran-1(3H)-one, together with four known sesquiterpenes, delta1(10)-aristolene-9beta-ol, debilon, nardosinone, and kanshone A, were isolated from the roots of Nardostachys chinensis. The structures of the new compounds were established on the basis of their spectroscopic data, and the structure of 1 was confirmed by X-ray crystallographic analysis. PMID- 16038568 TI - Limnophilaspiroketone, a highly oxygenated phenolic derivative from Limnophila geoffrayi. AB - A highly oxygenated phenolic spiroketone, limnophilaspiroketone (1), was isolated from the aerial parts of Limnophila geoffrayi collected in Thailand. The structure of 1 was determined based on spectroscopic data interpretation. This novel isolate, obtained as a major secondary metabolite constituent, was verified as a racemate using the Mosher ester method. PMID- 16038571 TI - Medication-related problems in the elderly : defining the issues and identifying solutions. AB - The elderly population is at great risk for medication-related problems as a result of age-related physiological changes, the presence of multiple chronic diseases and conditions, and the types and numbers of prescription and nonprescription medications they consume.Medication-related problems have received international attention. In the US, as many as 200,000 people may die of medication-related problems each year. Studies from other developed nations where patients have access to medications have determined that various types of medication-related problems occur in the elderly population in health care environments ranging from nursing homes to community dwelling settings.A variety of initiatives have been developed in an attempt to reduce the likelihood of medication-related problems in the elderly. The role that a pharmacist can play in identifying and preventing the negative health outcomes and costs of medication-related problems is being increasingly recognised. Indeed, numerous studies have now documented the positive impact that pharmacists can have in this area, and ongoing research is being conducted. With the growing numbers of elderly, and the increase in the numbers and types of medications available, it is imperative that awareness of medication-related problems be increased and that further initiatives be developed to optimise the positive outcomes of drug therapy in the elderly. PMID- 16038570 TI - Pharmacokinetic considerations relating to tacrolimus dosing in the elderly. AB - Relaxation of the upper age limits for solid organ transplantation coupled with improvements in post-transplant survival have resulted in greater numbers of elderly patients receiving immunosuppressant drugs such as tacrolimus. Tacrolimus is a potent agent with a narrow therapeutic window and large inter- and intraindividual pharmacokinetic variability. Numerous physiological changes occur with aging that could potentially affect the pharmacokinetics of tacrolimus and, hence, patient dosage requirements. Tacrolimus is primarily metabolised by cytochrome P450 (CYP) 3A enzymes in the gut wall and liver. It is also a substrate for P-glycoprotein, which counter-transports diffused tacrolimus out of intestinal cells and back into the gut lumen. Age-associated alterations in CYP 3A and P-glycoprotein expression and/or activity, along with liver mass and body composition changes, would be expected to affect the pharmacokinetics of tacrolimus in the elderly. However, interindividual variation in these processes may mask any changes caused by aging. More investigation is needed into the impact aging has on CYP and P-glycoprotein activity and expression. No single dose, intense blood-sampling study has specifically compared the pharmacokinetics of tacrolimus across different patient age groups. However, five population pharmacokinetic studies, one in kidney, one in bone marrow and three in liver transplant recipients, have investigated age as a co-variate. None found a significant influence for age on tacrolimus bioavailability, volume of distribution or clearance. The number of elderly patients included in each study, however, was not documented and may have been only small. It is likely that inter and intraindividual pharmacokinetic variability associated with tacrolimus increase in elderly populations. In addition to pharmacokinetic differences, donor organ viability, multiple co-morbidity, polypharmacy and immunological changes need to be considered when using tacrolimus in the elderly. Aging is associated with decreased immunoresponsiveness, a slower body repair process and increased drug adverse effects. Elderly liver and kidney transplant recipients are more likely to develop new-onset diabetes mellitus than younger patients. Elderly transplant recipients exhibit higher mortality from infectious and cardiovascular causes than younger patients but may be less likely to develop acute rejection. Elderly kidney recipients have a higher potential for chronic allograft nephropathy, and a single rejection episode can be more devastating. There is a paucity of information on optimal tacrolimus dosage and target trough concentration in the elderly. The therapeutic window for tacrolimus concentrations may be narrower. Further integrated pharmacokinetic pharmacodynamic studies of tacrolimus are required. It would appear reasonable, based on current knowledge, to commence tacrolimus at similar doses as those used in younger patients. Maintenance dose requirements over the longer term may be lower in the elderly, but the increased variability in kinetics and the variety of factors that impact on dosage suggest that patient care needs to be based around more frequent monitoring in this age group. PMID- 16038572 TI - The role of topical corticosteroids in bullous pemphigoid in the elderly. AB - Bullous pemphigoid (BP) is the most common autoimmune blistering skin disease. Until recently the condition was treated with oral corticosteroids. However, high dose corticosteroids are poorly tolerated in the elderly and their use has probably contributed to the high mortality rates observed in several cohorts. Accordingly, considerable effort has been directed at identifying corticosteroid sparing agents, such as immunosuppressant drugs, plasma exchange techniques, intravenous immunoglobulins and tetracycline, that can be used in this clinical setting. Many of these options have appeared to be useful in open series, but they have been found to be ineffective or only marginally effective when tested in randomised, controlled trials. An important breakthrough occurred with the demonstration in a large, randomised trial that a 'super-potent' topical corticosteroid (clobetasol propionate) was not only associated with a significant decrease in severe complications and mortality of BP patients, but was also more effective than oral prednisone. New strategies for BP should include topical clobetasol propionate as first-line therapy in the elderly and consider adjuvant therapy only in the very rare cases that are either resistant or intolerant to this treatment. PMID- 16038573 TI - Respiratory syncytial virus infection in elderly adults. AB - Respiratory syncytial virus (RSV) infection is now recognised as a significant problem in elderly adults. Epidemiological evidence indicates the impact of RSV in older adults may be similar to non-pandemic influenza, both in the community and in long-term care facilities. Attack rates in nursing homes are approximately 5-10% per year with significant rates of pneumonia (10-20%) and death (2-5%). Estimates using US health care databases and viral surveillance results over a 9 year period indicate that RSV infection causes approximately 10,000 all-cause deaths annually among persons >64 years of age. In contrast, influenza A accounted for approximately 37,000 yearly deaths in the same age group. The clinical features of RSV infection may be difficult to distinguish from those of influenza but include nasal congestion, cough, wheezing and low-grade fever. Older persons with underlying heart and lung disease and immunocompromised patients are at highest risk for RSV infection-related pneumonia and death. Diagnosis of RSV infection in adults is difficult because viral culture and antigen detection are insensitive, presumably because of low viral titres. The combination of serology and reverse transcriptase polymerase chain reaction assay offers the best sensitivity and specificity for the diagnosis of RSV but unfortunately these techniques are not widely available; consequently, most adult RSV disease goes unrecognised. Although treatment of RSV infection in the elderly is largely supportive, early therapy with ribavirin and intravenous gamma globulin improves survival in immunocompromised persons. An effective RSV vaccine has not yet been developed. Therefore, prevention of RSV is limited to standard infection control practices, such as hand washing and the use of gowns and gloves. PMID- 16038574 TI - Dysregulation of T-cell function in the elderly : scientific basis and clinical implications. AB - The function of the immune system is to maintain body integrity by defending against infections, cancers, autoimmune diseases and inflammation-related chronic diseases. The immune response is known to become defective with aging, leading to decreased longevity and appearance of age-related disease. The most important changes occur in T-cell immunity, and are manifested particularly as altered clonal expansion of cells of limited antigen specificity. The causes of these alterations are multifactorial, and include thymic involution, T-cell subset changes and signal transduction alterations. The clinical consequences of these changes are not well defined, except for their extremely important negative impact on defence against infections, especially by new pathogens, and decreased responses to vaccination. Considering the public health consequences of decreased immune competence in old age, strategies for immune response modulation are desirable to decrease the health burden for the elderly and improve their quality of life. PMID- 16038575 TI - Effect of divalproex sodium on behavioural and cognitive problems in elderly dementia. AB - BACKGROUND: Behavioural and psychological symptoms of dementia (BPSD) occur in up to 90% of individuals with dementia at some point in their illness. BPSD reduce patient quality of life, cause great distress to caregivers and are the most common reason for institutionalisation. In nursing homes, pharmacological measures (usually antipsychotics or benzodiazepines) are often required to control agitation and aggression in patients with dementia. However, no medications have been approved by the US Food and Drug Administration for this indication as yet. The antiepileptic agent divalproex sodium may have advantages in this setting because of lower rates of drug interactions and adverse effects in this patient population. OBJECTIVE: The aim of the study was to assess the impact of treatment with divalproex sodium on behavioural, mood and cognitive measures in a population of elderly nursing home residents with a history of behaviour problems associated with dementia. MATERIALS AND METHODS: The study was a retrospective analysis of a long-term care database which allowed assessment of the impact of divalproex sodium therapy on behavioural, mood and cognitive measures in elderly nursing home residents with a history of dementia-related behaviour problems. Minimum Data Set items relating to problems of behaviour, cognition and mood were collected prior to and after divalproex sodium treatment over a 1-year period. Two-phase generalised linear regression, with fixed intersections at the time of divalproex sodium initiation, was used to estimate trends in each measure prior to and after divalproex sodium initiation. Monotherapy, combination therapy with benzodiazepines and antipsychotics, and dose comparisons of divalproex sodium were studied. RESULTS: In all three situations (i.e. as monotherapy, in combination with benzodiazepines and antipsychotics, and at both higher and lower doses), divalproex sodium therapy was shown to have multiple beneficial effects on various behavioural, mood and cognition indicators in elderly nursing home residents. In general, the data seemed to support more favourable results for the higher divalproex sodium dose group. CONCLUSIONS: These data support the use of divalproex sodium in elderly nursing home residents with a history of dementia and behaviour problems and warrant conduct of prospective, randomised trials of the drug in this setting. PMID- 16038577 TI - Quinone-sensitized steady-state photolysis of acetophenone oximes under aerobic conditions: kinetics and product studies. AB - Oxidation of oximes via photosensitized electron transfer (PET) results in the formation of the corresponding ketones as the major product via oxime radical cations and iminoxyl radicals. The influence of electron-releasing and electron accepting substituents on these reactions was studied. The observed substituent effect strongly supports formation of iminoxyl radicals from the oximes via an electron transfer-proton transfer sequence rather than direct hydrogen atom abstraction. Correlation of the relative conversion of the oximes with Hammett parameters shows that radical effects dominate for the meta-substituted acetophenone oximes (rho(rad)/rho(pol) = 5.4; r2 = 0.93), whereas the para substituted oximes are influenced almost equally by radical and ionic effects (rho(rad)/rho(pol) = -1.1; r2 = 0.98). From these data sets we conclude that the follow-up reactions proceed through a number of intermediates with both radical and ionic character. This was confirmed by product studies with the use of an isotopically labeled nucleophile. In addition to the major oxidation product (ketone), a chlorine-containing product was often identified as well. Studies on the formation of this product show that the most likely pathway is either via a direct nucleophilic addition in a complex formed between the oxime radical cation and the chloranil radical anion or via a radical substitution (SH2) mechanism. These studies show that with the increasing use of oximes as drugs and pesticides, intake of these chemicals followed by enzymatic oxidation may result in the formation of a variety of reactive intermediates, which may lead to cell and tissue damage. PMID- 16038578 TI - Alkali ion exchanged Nafion as a confining medium for photochemical reactions. AB - Methanol-swollen Nafion beads were used as microreactors to control the photochemical reaction pathways. Product selectivity in three unimolecular reactions, namely, the photo-Fries rearrangement of naphthyl esters, Norrish Type I reaction of 1-phenyl-3-p-tolyl-propan-2-one and Norrish Type I and Type II reactions of benzoin alkyl ethers were examined. The influence of cations over the photodimerization of acenaphthylene and cross-photodimerization between acenaphthylene and N-benzyl maleimide included within Nafion were also examined. The photochemical behaviors of the above substrates were significantly altered within Nafion compared with their solution photochemistry. Of particular interest, the product distributions were found to depend on the counter cations of Nafion. PMID- 16038579 TI - Effects of crude and partially purified extracts from UV-B-irradiated rice leaves on Helicoverpa armigera (Hubner). AB - The effect of crude and partially purified extracts from ultraviolet-B (UV-B) irradiated rice (Oryza sativa L.) leaves on the growth and development of corn earworm, Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae), was investigated. Fifty muL droplets of a liquid diet containing different concentrations of the crude and partially purified extracts were fed to H. armigera neonates to determine possible short-term toxicity effects. A choice test using a solid artificial diet was also performed to determine larval feeding preferences and antifeedant effects. To study effects on the life history of the insect, different concentrations of the crude and partially purified extracts were also incorporated in the artificial diet and fed to individually confined neonates of H. armigera. The neonates were reared up to the adult stage. Results showed that crude and partially purified extracts of UV-B-irradiated rice leaves demonstrated antifeedant, growth-inhibitory and antibiotic properties against H. armigera. At high concentrations, the extract initially stimulated larval feeding; however, there were subsequent negative effects on pupal and adult traits, thereby reducing the reproductive potential of adults. These partially purified extracts appeared to have an antifertility effect because adults laid fewer eggs and, of those eggs laid, viability was lower. These results suggest that the accumulated flavonoids or other phenolics in UV-B-irradiated leaves, extracted from UV-B-resistant rice cultivar 'M202,' affected the growth, development and reproduction of H. armigera, a polyphagous insect pest. PMID- 16038580 TI - Comparisons of photo-fries rearrangements of 4-dodecylphenyl phenylacetate and two structurally related esters in hexane and polyethylene cages. How important are anchoring chains? AB - Photo-Fries rearrangements of 4-dodecylphenyl phenylacetate have been investigated in polyethylene films with 0-71% crystallinity and in hexane over a range of temperatures. The results are compared to those reported previously from phenyl phenylacetate and 1-naphthyl tetradecanoate to assess the influence of a long alkyl chain on the in-cage motions of the intermediate singlet radical pairs. It is demonstrated that the reactivity and selectivity of intimate singlet radical pairs can be tuned by judicious placement of long-chain substituents and selection of a specific polyethylene type as the reaction matrix. PMID- 16038581 TI - Photochemistry of 4-(2-nitrophenyl)-1,4-dihydropyridines. Evidence for electron transfer and formation of an intermediate. AB - New evidence about the path followed in the photochemical reaction of 4-(2 nitrophenyl)-1,4-dihydropyridines such as the drugs nifedipine (Compound 1) and nisoldipine (Compound 2) to give the corresponding nitrosophenylpyridines has been found through determination of the steady-state photochemical parameters and a comparison of the photoreactions in solution and in matrix at 90 K. Additional support is given by comparison with the isomeric 4-(3-nitrophenyl)dihydropyridine as well as with simpler derivatives, such as the corresponding 4 methyldihydropyridine. In Compounds 1 and 2, the lowest lying singlet, localized on the dihydropyridine chromophore, is deactivated by (largely exothermic) electron transfer to the nitrobenzene moiety, as evidenced by the complete quenching of the blue fluorescence observed in analogues not containing the electron-accepting group. Intramolecular proton transfer ensues in the 2 nitrophenyl derivatives with a relatively medium-independent quantum yield of approximately 0.3 and leads to an aromatic zwitterion, which is detected in matrix at 90 K (photoionization of this intermediate takes place in 2 methyltetrahydrofuran secondary). The intermediate is smoothly converted into the end product upon melting the glass. The 3-nitrophenyl analog, for which such a path is not available, is less reactive by about three orders of magnitude at 366 nm, although the quantum yield arrives at approximately 0.01 by irradiation at 254 nm in MeOH, reasonably via the nitrophenyl localized triplet. PMID- 16038582 TI - Mayak worker study: an improved biokinetic model for reconstructing doses from internally deposited plutonium. AB - The plutonium production facility known as the Mayak Production Association was put into operation in June 1948. A high incidence of cancer in the Mayak workers has been related to the level of exposure to plutonium, but uncertainties in tissue doses have hampered development of dose-risk relationships. As part of an effort to improve dose estimates for these workers, the systemic biokinetic model for plutonium currently recommended by the International Commission on Radiological Protection (ICRP) has been modified to reflect recently developed data and facilitate interpretation of case-specific information. This paper describes the proposed model and discusses its implications for dose reconstruction for the Mayak workers. PMID- 16038583 TI - Lung fibrosis in plutonium workers. AB - There have been few systematic studies of the non-malignant health effects of alpha-particle radiation in humans. Animal studies and a report on plutonium exposed workers from Russia suggest an association between high doses to the lung from plutonium exposure and the development of fibrotic lung disease. Prompted by a case of lung fibrosis in a retired plutonium worker, we tested the hypothesis that plutonium inhalation increases the risk for developing chest radiograph abnormalities consistent with pulmonary fibrosis. We conducted a retrospective study of nuclear weapons workers that included estimating absorbed doses to the lung with an internal dosimetry model. Our study population consisted of 326 plutonium-exposed workers with absorbed lung doses from 0 to 28 Sv and 194 unexposed workers. We compared the severity of chest radiograph interstitial abnormalities between the two groups using the International Labour Organization profusion scoring system. There was a significantly higher proportion of abnormal profusion scores among plutonium-exposed workers (17.5%) than among unexposed workers (7.2%), P < 0.01. Lung doses of 10 Sv or greater conferred a 5.3-fold risk (95% CI 1.2-23.4) of having an abnormal chest X ray consistent with pulmonary fibrosis when compared with unexposed individuals after controlling for the effects of age, smoking and asbestos exposure. This study shows that plutonium may cause lung fibrosis in humans at absorbed lung doses above 10 Sv. PMID- 16038584 TI - Influence of polymorphisms in DNA repair genes XPD, XRCC1 and MGMT on DNA damage induced by gamma radiation and its repair in lymphocytes in vitro. AB - DNA single-strand breaks (SSBs) were quantified by single-cell gel electrophoresis and micronucleated and apoptotic cells were quantified by microscopic assays in peripheral blood lymphocytes after irradiation on ice with 2 Gy of 60Co gamma radiation, and their association with polymorphisms of genes that encode proteins of different DNA repair pathways and influence cancer risk (XPD codon 312Asp --> Asn and 751Lys --> Gln, XRCC1 399Arg --> Gln, and MGMT 84Leu --> Phe) was studied. In unirradiated lymphocytes, SSBs were significantly more frequent in individuals older than the median age (52 years) (P = 0.015; n = 81), and the frequency of apoptotic or micronucleated cells was higher in individuals with alleles coding for Asn at XPD 312 or Gln at 751 (P = 0.030 or 0.023 ANOVA, respectively; n = 54). The only polymorphism associated with the background SSB level was MGMT 84Phe (P = 0.04, ANOVA; n = 66). After irradiation, SSB levels and repair parameters did not differ significantly with age or smoking habit. The SSB level varied more than twofold and the repair rate and level of unrepaired SSBs more than 10-fold between individuals. The presence of variant alleles coding for Asn at XPD 312 was associated with more radiation-induced SSBs (P = 0.014) and fewer unrepaired SSBs (P = 0.008), and the phenotype (> median induced SSBs/< median unrepaired SSBs) was seen in the majority of XPD 312Asn/Asn homozygotes; the odds ratio for variant homozygotes to show this phenotype was 5.2 (95% confidence interval 1.4-19.9). The hypothesis is discussed that XPD could participate in repair of ionizing radiation-induced DNA damage. While it cannot be excluded that the effects observed are due to cosegregating polymorphisms or that the responses of lymphocytes are not typical of other cell types, the results suggest that polymorphism of DNA repair genes, particularly XPD, is one factor implicated in the variability of responses to ionizing radiation between different individuals. PMID- 16038585 TI - Role of homologous recombination in the alpha-particle-induced bystander effect for sister chromatid exchanges and chromosomal aberrations. AB - The bystander effect for sister chromatid exchanges (SCEs) and chromosomal aberrations was examined in hamster cell lines deficient in either DNA-PKcs (V3 cells, deficient in nonhomologous end joining, NHEJ) or RAD51C (irs3 cells, deficient in homologous recombination, HR). Cells synchronized in G0/G1 phase were irradiated with very low fluences of alpha particles such that < 1% of the nuclei were traversed by an alpha particle. Wild-type cells showed a prominent bystander response for SCE induction; an even greater effect was observed in V3 cells. On the other hand, no significant induction of SCE was observed in the irs3 RAD51C-deficient bystander cells irradiated at various stages in the cell cycle. Whereas a marked bystander effect for chromosomal aberrations occurred in V3 cells, the induction of chromosomal aberrations in irs3 bystander cells was minimal and similar to that of wild-type cells. Based on these findings, we hypothesize that HR is essential for the induction of SCE in bystander cells; however, HR is unable to repair the DNA damage induced in NHEJ-deficient bystander cells that leads to either SCE or chromosomal aberrations. PMID- 16038586 TI - Genotoxicity of neutrons in Drosophila melanogaster. Somatic mutation and recombination induced by reactor neutrons. AB - This paper describes the observation of a direct relationship between the absorbed doses of neutrons and the frequencies of somatic mutation and recombination using the wing somatic mutation and recombination test (SMART) of Drosophila melanogaster. This test was used for evaluating the biological effects induced by neutrons from the Triga Mark III reactor of Mexico. Two different reactor power levels were used, 300 and 1000 kW, and two absorbed doses were tested for each power level: 1.6 and 3.2 Gy for 300 kW and 0.84 and 1.7 Gy for 1000 kW. A linear relationship was observed between the absorbed dose and the somatic mutation and recombination frequencies. Furthermore, these frequencies were dependent on larval age: In 96-h-old larvae, the frequencies were increased considerably but the sizes of the spots were smaller than in 72-h-old larvae. The analysis of the balancer-heterozygous progeny showed a linear absorbed dose- response relationship, although the responses were clearly lower than found in the marker-trans-heterozygous flies. Approximately 65% of the genotoxicity observed is due to recombinational events. The results of the study indicate that thermal and fast neutrons are both mutagenic and recombinagenic in the D. melanogaster wing SMART, and that the frequencies are dependent on neutron dose, reactor power, and the age of the treated larvae. PMID- 16038587 TI - The heat-shock factor is not activated in mammalian cells exposed to cellular phone frequency microwaves. AB - There has been considerable interest in the biological effects of exposure to radiofrequency electromagnetic radiation, given the explosive growth of cellular telephone use, with the possible induction of malignancy being a significant concern. Thus the determination of whether nonthermal effects of radiofrequency electromagnetic radiation contribute to the process leading to malignancy is an important task. One proposed pathway to malignancy involves the induction of the stress response by exposures to cell phone frequency microwaves. The first step in the induction of the stress response is the activation of the DNA-binding activity of the specific transcription factor involved in this response, the heat shock factor (HSF). The DNA-binding activity of HSF in hamster, mouse and human cells was determined after acute and continuous exposures to frequency domain multiple access (FDMA)- or code domain multiple access (CDMA)-modulated microwaves at low (0.6 W/kg) or high (approximately 5 W/kg) SARs at frequencies used for mobile communication. The DNA-binding activity of HSF was monitored using a gel shift assay; the calibration of this assay indicated that an increase of approximately 10% in the activation of the DNA-binding activity of HSF after a 1 degrees C increase in temperature could be detected. We failed to detect any increase in the DNA-binding ability of HSF in cultured mammalian cells as a consequence of any exposure tested, within the sensitivity of our assay. Our results do not support the notion that the stress response is activated as a consequence of exposure to microwaves of frequencies associated with mobile communication devices. PMID- 16038588 TI - DNA interduplex crosslinks induced by Al(Kalpha) X rays under vacuum. AB - Dry pGEM-3Zf(-) plasmid DNA was exposed to Al(kalpha) X rays (1.5 keV) for various times in an ultra-high vacuum chamber with mean absorbed dose rates ranging from 1.8 to 41.7 Gy s(-1). The different forms of plasmid DNA were separated by neutral agarose gel electrophoresis and quantified by staining and laser scanning. In addition to the bands for supercoiled, nicked circular, linear and concatameric forms of plasmid DNA, two additional bands were observed in X irradiated samples; these migrated at rates similar to those for 8-kb and >10-kb linear double-stranded DNA. Digestion of irradiated DNA with the restriction enzymes EcoR1 and PvuI suggested that the two slowly migrating bands were interduplex crosslinked DNA. Alkaline agarose gel electrophoresis of irradiated DNA digested with EcoR1 confirmed that the interduplex crosslink was covalent. Exposure-response curves were determined for the formation of nicked circular, linear and interduplex crosslinked DNA as well as for the loss of supercoiled and concatameric DNA. Formation and loss of these species were independent of absorbed dose rate over a 20-fold range. The G values for DNA single-strand breaks, double-strand breaks and crosslinks were determined to be 62 +/- 6, 5.6 +/- 0.6 and 16 +/- 4 nmol J(-1), respectively. The formation of DNA interduplex crosslinks appears to be due to single event. The mechanism responsible for the formation of DNA interduplex crosslinks is discussed with emphasis on its implications in vivo. PMID- 16038589 TI - Monte Carlo simulation of base and nucleotide excision repair of clustered DNA damage sites. I. Model properties and predicted trends. AB - DNA is constantly damaged through endogenous processes and by exogenous agents, such as ionizing radiation. Base excision repair (BER) and nucleotide excision repair (NER) help maintain the stability of the genome by removing many different types of DNA damage. We present a Monte Carlo excision repair (MCER) model that simulates key steps in the short-patch and long-patch BER pathways and the NER pathway. The repair of both single and clustered damages, except double-strand breaks (DSBs), is simulated in the MCER model. Output from the model includes estimates of the probability that a cluster is repaired correctly, the fraction of the clusters converted into DSBs through the action of excision repair enzymes, the fraction of the clusters repaired with mutations, and the expected number of repair cycles needed to completely remove a clustered damage site. The quantitative implications of alternative hypotheses regarding the postulated repair mechanisms are investigated through a series of parameter sensitivity studies. These sensitivity studies are also used to help define the putative repair characteristics of clustered damage sites other than DSBs. PMID- 16038590 TI - Monte carlo simulation of base and nucleotide excision repair of clustered DNA damage sites. II. Comparisons of model predictions to measured data. AB - Clustered damage sites other than double-strand breaks (DSBs) have the potential to contribute to deleterious effects of ionizing radiation, such as cell killing and mutagenesis. In the companion article (Semenenko et al., Radiat. Res. 164, 180-193, 2005), a general Monte Carlo framework to simulate key steps in the base and nucleotide excision repair of DNA damage other than DSBs is proposed. In this article, model predictions are compared to measured data for selected low-and high-LET radiations. The Monte Carlo model reproduces experimental observations for the formation of enzymatic DSBs in Escherichia coli and cells of two Chinese hamster cell lines (V79 and xrs5). Comparisons of model predictions with experimental values for low-LET radiation suggest that an inhibition of DNA backbone incision at the sites of base damage by opposing strand breaks is active over longer distances between the damaged base and the strand break in hamster cells (8 bp) compared to E. coli (3 bp). Model estimates for the induction of point mutations in the human hypoxanthine guanine phosphoribosyl transferase (HPRT) gene by ionizing radiation are of the same order of magnitude as the measured mutation frequencies. Trends in the mutation frequency for low- and high LET radiation are predicted correctly by the model. The agreement between selected experimental data sets and simulation results provides some confidence in postulated mechanisms for excision repair of DNA damage other than DSBs and suggests that the proposed Monte Carlo scheme is useful for predicting repair outcomes. PMID- 16038591 TI - A complete dielectric response model for liquid water: a solution of the Bethe ridge problem. AB - We present a complete yet computationally simple model for the dielectric response function of liquid water over the energy-momentum plane, which, in contrast to earlier models, is consistent with the recent inelastic X-ray scattering spectroscopy data at both zero and finite momentum transfer values. The model follows Ritchie's extended-Drude algorithm and is particularly effective at the region of the Bethe ridge, substantially improving previous models. The present development allows for a more accurate simulation of the inelastic scattering and energy deposition process of low-energy electrons in liquid water and other biomaterials. As an example, we calculate the stopping power of liquid water for electrons over the 0.1-10 keV range where direct experimental measurements are still impractical and the Bethe stopping formula is inaccurate. The new stopping power values are up to 30-40% lower than previous calculations. Within the range of validity of the first Born approximation, the new values are accurate to within the experimental uncertainties (a few percent). At the low end, the introduction of Born corrections raises the uncertainty to perhaps approximately 10%. Thus the present model helps extend the ICRU electron stopping power database for liquid water down to about two orders of magnitude with a comparable level of uncertainty. PMID- 16038592 TI - Epithermal neutron beams for clinical studies of boron neutron capture therapy: a dosimetric comparison of seven beams. AB - A comparison of seven epithermal neutron beams used in clinical studies of boron neutron capture therapy (BNCT) in Sweden (Studsvik), Finland (Espoo), Czech Republic (ReZ), The Netherlands (Petten) and the U.S. (Brookhaven and Cambridge) was performed to facilitate sharing of preclinical and clinical results. The physical performance of each beam was measured using a common dosimetry method under conditions pertinent to brain irradiations. Neutron fluence and absorbed dose measurements were performed with activation foils and paired ionization chambers on the central axis both in air and in an ellipsoidal water phantom. The overall quality of each beam was assessed by figures of merit determined from the total weighted dose profiles that assumed the presence of boron in tissue. The in air specific beam contamination from both fast neutrons and gamma rays ranged between 8 and 65 x 10(-11) cGy(w) cm2 for the different beams and the epithermal neutron flux intensities available at the patient position differed by more than a factor of 20 from 0.2-4.3 x 10(9) n cm(-2) s(-1). Percentage depth dose profiles measured in-phantom for the individual photon, thermal and fast-neutron dose components differed only subtly in shape between facilities. Assuming uptake characteristics consistent with the currently used boronated phenylalanine, all the epithermal beams exhibit a useful penetration of 8 cm or greater that is sufficient to irradiate a lesion seated at the brain midline. The performance of the existing facilities will benefit from the introduction of advanced compounds through improved beam penetrability. This could increase by as much as 2 cm for the purest of beams, although the beam intensities generally need to be increased to between 2-5 x 10(9) n cm(-2) s(-1) to maintain manageable irradiation times. These data provide the first consistent measurement of beam performance at the different centers and will enable a preliminary normalization of the calculated patient dosimetry. PMID- 16038593 TI - The MIT user center for neutron capture therapy research. AB - Neutron capture therapy (NCT) research encompasses a wide range of preclinical and clinical studies needed to develop this promising but complex cancer treatment. Many specialized facilities and capabilities including thermal and epithermal neutron irradiation facilities, boron analysis, specialized mixed field dosimetry, animal care facilities and protocols, cell culture laboratories, and, for human clinical studies, licenses and review board approvals are required for NCT research. Such infrastructure is essential, but much of it is not readily available within the community. This is especially true for neutron irradiation facilities, which often require significant development and capital investment too expensive to duplicate at each site performing NCT research. To meet this need, the NCT group at the Massachusetts Institute of Technology (MIT) has established a User Center for NCT researchers that is already being accessed successfully by various groups. This paper describes the facilities, capabilities and other resources available at MIT and how the NCT research community can access them. PMID- 16038594 TI - Intracellular sequestration of 223Ra by the iron-storage protein ferritin. AB - Incorporation of bone-seeking, alpha-particle-emitting, heavy-metal radionuclides dramatically increases the incidence of osteosarcoma in humans and experimental animals. The accumulation of these radionuclides within the mineral phase of the bone matrix is believed to result in local irradiation of only those proliferating cells close to the bone surface. We now present evidence for a more general pathway for the irradiation of target cells, mediated through the sequestration of heavy-metal radionuclides by the intracellular iron-storage protein ferritin. In vitro studies reveal the transfer of radionuclide from a 223Ra-transferrin complex into immunoprecipitable cytosolic ferritin. In vivo studies confirm the co-localization of incorporated 224Ra and cellular iron stores. This pathway would result in the highly localized irradiation of ferritin containing cells. Since osteoblastic cells express large quantities of a ferritin isoform specialized in long-term metal storage, we suggest that this may represent an unrecognized source of intracellular irradiation by alpha-particle emitting radionuclides. Such a local concentration within target cells has implications both for cellular dosimetry and for inferences of track length and target cell populations within the skeleton. PMID- 16038595 TI - Introduction: the burden of bipolar depression. PMID- 16038596 TI - The impact of bipolar depression. AB - Bipolar disorder is a chronic, intermittent illness that is associated with high morbidity and mortality. In addition, patients with bipolar disorder often have comorbid psychiatric conditions (such as anxiety disorders, alcohol or substance abuse, and eating disorders) or medical disorders (such as obesity), which result in increased burden of illness for the patients, family members, and treating clinicians. Although bipolar disorder consists of recurring episodes of mania and depression, patients spend more time depressed than manic. Bipolar depression is associated with a greater risk of suicide and of impairment in work, social, or family life than mania. This health burden also results in direct and indirect economic costs to the individual and society at large. Bipolar depression is often undiagnosed or misdiagnosed as unipolar depression, resulting in incorrect or inadequate treatment. Available treatments for bipolar depression include medications such as lithium, selected anticonvulsants, and the atypical antipsychotics. Traditional antidepressants are not recommended as monotherapy for bipolar depression as they can induce switching to mania. Early and accurate diagnosis, aggressive management, and earlier prophylactic treatment regimens are needed to overcome the impact of depressive episodes in patients with bipolar disorder. PMID- 16038597 TI - Challenges in the management of bipolar depression. AB - Bipolar depression has started to receive more attention in clinical trials only relatively recently, despite the fact that patients spend more time in the depressed phase than in the manic phase of bipolar disorder. The diagnosis and management of bipolar depression are challenging, and many patients are undiagnosed or misdiagnosed due to symptom similarities with unipolar depression or other illnesses and/or comorbidities. Untreated or inappropriately treated bipolar depression adds to the burden of illness and is associated with a greater risk of suicide. Treatment options include lithium, lamotrigine, atypical antipsychotics, and traditional antidepressants, such as the selective serotonin reuptake inhibitors. However, traditional antidepressants are recommended with caution due to their potential risk of switching patients into mania. Some atypical antipsychotics have shown efficacy in bipolar depression, although longer-term studies are warranted. The choice of treatment for different subgroups of patients with bipolar depression, including those with comorbid anxiety, may vary and also needs further study. Other important issues that require further investigation include the recognition of the core features of bipolar depression and the threshold symptoms for treatment, as well as the optimal treatment choices for monotherapy or combination therapy, and acute versus long-term management of bipolar depression. PMID- 16038598 TI - Reevaluating therapies for bipolar depression. AB - The most commonly employed pharmacotherapies for bipolar depression include antidepressants, lithium, and anticonvulsants, such as lamotrigine, valproate, and carbamazepine. A combination of these agents, usually an antidepressant and a mood stabilizer, is often required to achieve an optimal response. However, some treatment guidelines still caution that antidepressant exposure should be minimized in patients with bipolar depression, due to concern that they may trigger treatment-emergent mania or cycle acceleration. This advice prevails despite data showing that antidepressants are effective in treating bipolar depression and evidence that coadministration of a mood-stabilizing medication, at least with modern antidepressants, such as the selective serotonin reuptake inhibitors, can reduce the risk of treatment-emergent mania to levels comparable with those observed with mood stabilizer monotherapy. Although the antidepressant efficacy of most mood stabilizers has not been satisfactorily proven, first-line therapy with 1 mood stabilizer alone or a combination of 2 mood stabilizers is still recommended by many guidelines. Inappropriate treatment of bipolar depression may leave patients at high risk of suicide and increased chronicity of symptoms; effective therapy should, therefore, be provided as early as possible. The efficacy and safety of antidepressants for bipolar depression both as monotherapy and when combined with a mood stabilizer should be studied in adequately powered trials in order to revise treatment guidelines. Electroconvulsive therapy remains an option for treatment-refractory patients and those intolerant to pharmacologic treatment, as well as patients who are pregnant or at high risk of suicide. PMID- 16038599 TI - Clinical highlights in bipolar depression: focus on atypical antipsychotics. AB - Despite the considerable burden of bipolar depression, the treatment of this debilitating phase of bipolar disorder is suboptimally addressed by currently available pharmacologic options. Consequently, there is a need for the development of new treatment options with enhanced efficacy and tolerability. Evidence of antidepressant efficacy for some of the atypical antipsychotics in the treatment of bipolar depression has recently emerged. The findings of a large scale, placebo-controlled, double-blind, randomized clinical study of olanzapine alone and in combination with fluoxetine, and a similar study of quetiapine monotherapy, suggest that some of the atypical antipsychotics may be efficacious in treating depressive symptoms in patients with bipolar I disorder. Subpopulation analyses suggest that quetiapine monotherapy and the olanzapine plus fluoxetine combination appear to be effective in treating depression in patients with a rapid-cycling course. The magnitude of improvement in depressive symptoms in the bipolar I population appears to be larger for quetiapine monotherapy compared with either olanzapine or olanzapine plus fluoxetine; however, the limitations of such a cross-study comparison are acknowledged. Both olanzapine monotherapy and combination therapy, as well as quetiapine monotherapy, were well tolerated. The overall incidence of treatment-emergent mania was low and comparable with placebo in both studies. Somnolence, weight gain, increased appetite and nonfasting glucose and cholesterol levels were more commonly reported in patients treated with olanzapine monotherapy or combination therapy compared with placebo. Dry mouth, sedation/somnolence, dizziness, and constipation were more commonly associated with quetiapine treatment. Large, controlled studies are needed to determine whether other psychotropic agents have antidepressant properties that would make them suitable for use in patients with bipolar depression. In addition, direct comparison of the regimens used in the current study should determine whether the differences evident between them can be confirmed. PMID- 16038600 TI - The package of care for patients with bipolar depression. AB - Although pharmacotherapy is the mainstay of the comprehensive program of medical care for the management of patients with bipolar disorder, the additional benefits of psychosocial interventions for the patient, family, and caregivers are now being recognized and increasingly adopted. Several facets of bipolar disorder can be addressed more effectively by instituting adjunctive psychosocial interventions. Recent clinical evidence indicates that combining pharmacotherapy with psychosocial interventions, which are tailored to patients' individual needs, may decrease the risk of relapse, improve patient adherence, and decrease the number and length of hospitalizations. A multidisciplinary approach may also enhance long-term patient outcomes such as mood stability, enhanced occupational and/or social functioning, and overall quality of life. Psychoeducation helps individuals become active and informed participants in the management of their illness, promoting a collaborative relationship between patients and their caregivers. However, psychosocial interventions are not useful for all patients with bipolar illness and may be more useful in addressing some problems than others. Evidence would suggest that psychosocial interventions, with continuing pharmacotherapy, are best used as prophylaxis and during periods of remission to prevent further episodes. Further randomized, clinical trials will help to define which components of psychosocial intervention are most effective in patients with bipolar disorder. PMID- 16038601 TI - Atypical antipsychotics in bipolar depression: potential mechanisms of action. AB - "Conventional" antidepressants, such as the selective serotonin reuptake inhibitors (SSRIs), bupropion, or serotonin-norepinephrine reuptake inhibitors, are not recommended as monotherapy for bipolar depression. Although they are likely to provide effective symptom relief in combination with mood stabilizers, the risk of precipitating a switch to mania often complicates their use even as combination therapy. Recently, 2 psychotropic medications approved for treating acute mania, olanzapine and quetiapine, have also been shown to possess antidepressant activity without destabilizing mood and, as such, are potential mood stabilizers. This article aims to review the mechanism of action of conventional antidepressants and newer agents that are effective in the treatment of bipolar depression. A number of mechanisms have been postulated to play a role in the effective treatment of bipolar depression, including targets as diverse as serotonin (5-HT), norepinephrine, dopamine, gamma-aminobutyric acid (GABA), glutamate, and various second messenger signaling pathways. A review of the data reveals an important point of commonality among the antidepressant treatments, olanzapine, and quetiapine. Antidepressant treatments, such as norepinephrine reuptake inhibitors, SSRIs, and electroconvulsive therapy, induce a reduction of 5-HT(2A) receptors. Both olanzapine and quetiapine not only are antagonists at this receptor but also induce downregulation of 5-HT(2A) receptors. It is possible that the antidepressant efficacy of these agents is mediated by this receptor, while the additional benefit of olanzapine and quetiapine over unimodal antidepressant treatments, in terms of stabilizing mood, may be provided by their concomitant dopamine D(2) antagonism. Further studies should be conducted to examine these hypotheses. PMID- 16038602 TI - Role of mitochondria in the mechanisms of glutamate toxicity. AB - Current data on glutamate-induced functional and morphological changes in mitochondria correlating with or being a result of their membrane potential changes are reviewed. The important role of Ca2+, Na+, and H+ in the potentiation of such changes is considered. It is assumed that glutamate-induced loss of mitochondrial potential is mediated by Ca2+ overload resulting in the induction of nonspecific permeability of the inner mitochondrial membrane. PMID- 16038603 TI - Reactive oxygen species and human inflammatory periodontal diseases. AB - Reactive oxygen species (ROS) have emerged as important signaling molecules in the regulation of various cellular processes. They can be generated by the mitochondrial electron transport chain in mitochondria and activation of polymorphonuclear leukocytes (PMN) during inflammatory conditions. Excessive generation of ROS may result in attack of and damage to most intracellular and extracellular components in a living organism. Moreover, ROS can directly induce and/or regulate apoptotic and necrotic cell death. Periodontal pathologies are inflammatory and degenerative diseases. Several forms of periodontal diseases are associated with activated PMN. Damage of tissues in inflammatory periodontal pathologies can be mediated by ROS resulting from the physiological activity of PMN during the phagocytosis of periodontopathic bacteria. PMID- 16038604 TI - Structure, properties, and probable physiological role of small heat shock protein with molecular mass 20 kD (Hsp20, HspB6). AB - This review is devoted to critical analysis of data concerning the structure and functions of small heat shock proteins with apparent molecular mass 20 kD (Hsp20). We describe the structure of Hsp20, its phosphorylation by different protein kinases, interaction of Hsp20 with other small heat shock proteins, and chaperone activity of Hsp20. The distribution of Hsp20 in different animal tissues and the factors affecting expression of Hsp20 are also described. Data on the possible involvement of Hsp20 in regulation of platelet aggregation and glucose transport are presented and analyzed. Special attention is paid to literature data describing probable regulatory effect of Hsp20 on contraction of smooth muscle. Two hypotheses postulating direct effect of Hsp20 on actomyosin interaction or its effect on cytoskeleton are compared and analyzed. The most recent data on the effect of Hsp20 on apoptosis and contractile activity of cardiomyocytes are also presented. PMID- 16038605 TI - Cloning and characterization of zebra fish SPATA4 gene and analysis of its gonad specific expression. AB - The spermatogenesis associated 4 gene (SPATA4, previously named TSARG2) was first cloned in human tissues and was reported to be a candidate spermatocyte apoptosis related gene that is expressed specifically in testis. Analysis of SPATA4 expression and regulation in zebra fish may provide insight into the understanding of the complicated process of gonadogenesis. In this study, we cloned and characterized the SPATA4 gene from zebra fish (Danio rerio), which is homologous to human and mouse SPATA4. Zebra fish SPATA4 consists of six exons separated by five introns, as all SPATA4 genes in vertebrates. A promoter region was predicted using homologous blast and cloned for further study, and possible transcription factors were analyzed in this region. The putative protein encoded by this gene was analyzed using bioinformatics methods. Multi-tissue RT-PCR results demonstrated that the zebra fish SPATA4 gene is expressed specifically in testis and slightly in ovary. Analysis of the SPATA4 sequence and its spatial expression pattern indicate that this gene is highly conserved and may play an important role in the process of zebra fish gonadogenesis. PMID- 16038606 TI - Use of a reverse micelle system for study of oligomeric structure of NAD+ reducing hydrogenase from Ralstonia eutropha H16. AB - Inclusion of an oligomeric enzyme, NAD+-dependent hydrogenase from the hydrogen oxidizing bacterium Ralstonia eutropha, into a system of reverse micelles of different sizes resulted in its dissociation into catalytically active heterodimers and subunits, which were characterized in reactions with various substrates. It was found that: 1) the native tetrameric form of this enzyme catalyzes all types of studied reactions; 2) hydrogenase dimer, HoxHY, is a minimal structural unit catalyzing hydrogenase reaction with an artificial electron donor, reduced methyl viologen; 3) all structural fragments containing FMN and NAD+/NADH-binding sites exhibit catalytic activity in diaphorase reactions with one- and two-electron acceptors; 4) small subunits, HoxY and HoxU also exhibit activity in diaphorase reactions with artificial acceptors. These results can be considered as indirect evidence that the second FMN molecule may be associated with one of the small subunits (HoxY or HoxU) of the hydrogenase from R. eutropha. PMID- 16038607 TI - Intracellular signal cascade in CD4+ T-lymphocyte migration stimulated by interferon-gamma-inducible protein-10. AB - The intracellular signal cascades involved in chemokine-stimulated migration of in vitro activated human peripheral blood CD4+ T-lymphocytes were investigated. IP-10-mediated chemotactic response of lymphocytes was decreased in the presence of selective inhibitors of Src-kinases (by 40-45%), PI3-kinases (35-40%), and MAP kinases ERK1/2 (35-40%) and p38 (20%). Combined addition of specific inhibitors of Src-kinases and PI3-kinases and inhibitors of Src-kinases and ERK1/2 MAP kinases did not result in the further increase of the inhibitory effect, while the combined addition of specific inhibitors of PI3-kinases and ERK1/2 MAP kinases decreased migration of CD4+ T-lymphocytes more effectively (by 55-60%) than any individual inhibitor. Immunoblotting analysis of activation of MAP kinases ERK1/2 and p38 revealed increased level of phosphorylation of ERK1/2 and p38 MAP-kinases in the presence IP-10. Selective inhibitors of Src-kinases and PI3-kinases significantly inhibited phosphorylation of p38 but did not influence phosphorylation of ERK1/2 MAP-kinases. Our results suggest that Src-kinases, PI3 kinases, and ERK1/2 MAP-kinases are involved in intracellular signal cascade activated during IP-10-stimulated migration of T-lymphocytes, whereas p38 MAP kinases do not participate in the migration process, although its activation induced by IP-10 depends on Src-kinases and PI3-kinases. PMID- 16038608 TI - New effective method for analysis of the component composition of enzyme complexes from Trichoderma reesei. AB - A method for analysis of the component composition of multienzyme complexes secreted by the filamentous fungus Trichoderma reesei was developed. The method is based on chromatofocusing followed by further identification of protein fractions according to their substrate specificity and molecular characteristics of the proteins. The method allows identifying practically all known cellulases and hemicellulases of T. reesei: endoglucanase I (EG I), EG II, EG III, cellobiohydrolase I (CBH I), CBH II, xylanase I (XYL I), XYL II, beta-xylosidase, alpha-L-arabinofuranosidase, acetyl xylan esterase, mannanase, alpha galactosidase, xyloglucanase, polygalacturonase, and exo-beta-1,3-glucosidase. The component composition of several laboratory and commercial T. reesei preparations was studied and the content of the individual enzymes in these preparations was quantified. The influence of fermentation conditions on the component composition of secreted enzyme complexes was revealed. The characteristic features of enzyme preparations obtained in "cellulase" and "xylanase" fermentation conditions are shown. PMID- 16038609 TI - Role of positively charged residues lys267, lys270, and arg411 of cytochrome p450scc (CYP11A1) in interaction with adrenodoxin. AB - Cytochrome P450scc and adrenodoxin are redox proteins of the electron transfer chain of the inner mitochondrial membrane steroid hydroxylases. In the present work site-directed mutagenesis of the charged residues of cytochrome P450scc and adrenodoxin, which might be involved in interaction, was used to study the nature of electrostatic contacts between the hemeprotein and the ferredoxin. The target residues for mutagenesis were selected based on the theoretical model of cytochrome P450scc-adrenodoxin complex and previously reported chemical modification studies of cytochrome P450scc. In the present work, to clarify the molecular mechanism of hemeprotein interaction with ferredoxin, we constructed cytochrome P450scc Lys267, Lys270, and Arg411 mutants and Glu47 mutant of adrenodoxin and analyzed their possible role in electrostatic interaction and the role of these residues in the functional activity of the proteins. Charge neutralization at positions Lys267 or Lys270 of cytochrome P450scc causes no significant effect on the physicochemical and functional properties of cytochrome P450scc. However, cytochrome P450scc mutant Arg411Gln was found to exhibit decreased binding affinity to adrenodoxin and lower activity in the cholesterol side chain cleavage reaction. Studies of the functional properties of Glu47Gln and Glu47Arg adrenodoxin mutants indicate that a negatively charged residue in the loop covering the Fe2S2 cluster, being important for maintenance of the correct architecture of these structural elements of ferredoxin, is not directly involved in electrostatic interaction with cytochrome P450scc. Moreover, our results indicate the presence of at least two different binding (contact) sites on the proximal surface of cytochrome P450scc with different electrostatic input to interaction with adrenodoxin. In the binary complex, the positively charged sites of the proximal surface of cytochrome P450scc well correspond to the two negatively charged sites of adrenodoxin: the "interaction" domain site and the "core" domain site. PMID- 16038610 TI - Cloning and molecular modeling of duodenase with respect to evolution of substrate specificity within mammalian serine proteases that have lost a conserved active-site disulfide bond. AB - Mammalian serine proteases such as the chromosome 14 (Homo sapiens, Mus musculus) located granzymes, chymases, cathepsin G, and related enzymes including duodenase collectively represent a special group within the chymotrypsin family which we refer to here as "granases". Enzymes of this group have lost the ancient active site disulfide bond Cys191-Cys220 (bovine chymotrypsinogen A numbering) which is strongly conserved in classic serine proteases such as pancreatic, blood coagulation, and fibrinolysis proteases and others (granzymes A, M, K and leukocyte elastases). We sequenced the cDNA encoding bovine (Bos taurus) duodenase, a granase with unusual dual trypsin-like and chymotrypsin-like specificity. The sequence revealed a 17-residue signal peptide and two-residue (GlyLys) activation peptide typical for granases. Production of the mature enzyme is apparently accompanied by further proteolytic processing of the C-terminal pentapeptide extension of duodenase. Similar C-terminal processing is known for another dual-specific granase, human cathepsin G. Using phylogenetic analysis based on 39 granases we retraced the evolution of residues 189 and 226 crucial for serine protease primary specificity. The analysis revealed that while there is no obvious link between mutability of residue 189 and the appearance of novel catalytic properties in granases, the mutability of residue 226 evidently gives rise to different specificity subgroups within this enzyme group. The architecture of the extended substrate-binding site of granases and structural basis of duodenase dual specificity based on molecular dynamic method are discussed. We conclude that the marked selectivity of granases that is crucial to their role as regulatory proteases has evolved through the fine-tuning of specificity at three levels--primary, secondary, and conformational. PMID- 16038611 TI - Isolation and characterization of biochemical properties of DNA methyltransferase FauIA modifying the second cytosine in the nonpalindromic sequence 5'-CCCGC-3'. AB - A gene encoding DNA methyltransferase (methylase) FauIA of the restriction modification system FauI from Flavobacterium aquatile (recognizing sequence 5' CCCGC-3') was cloned in pJW vector. The latter was used for transformation of E. coli RRI cells followed by subsequent thermoinduction and biomass elaboration. Highly purified DNA methyltransferase FauIA preparation was obtained using chromatography on different sorbents. The molecular mass of the isolated enzyme of about 39 kD corresponds to its theoretical value. The enzyme was characterized by temperature and pH optima of 33 degrees C and pH 7.5, respectively. Methylation of a synthetic oligonucleotide by FauIA methylase followed by its cleavage with various restrictases and analysis of the resultant restriction fragments revealed that FauIA methylase modified the second cytosine residue in the sequence 5'-CCCGC-3'. Kinetic analysis revealed Km and catalytic constant values of 0.16 microM and 0.05 min(-1), respectively. PMID- 16038612 TI - Calcium homeostasis change in CD4+ T lymphocytes from human peripheral blood during differentiation in vivo. AB - Resting naive CD4+(CD45R0-)CD45RA+ T cells are sensitive to ionomycin. In contrast, resting CD4+(CD45RA-)CD45R0+ memory T cells show resistance to this Ca2+ ionophore. In the present study, the ability of activated T lymphocytes to respond to ionomycin during the transition from naive precursors into memory T cells has been analyzed. Activated CD4+(CD45RA+)CD45R0+ T cells are always present both in human peripheral blood (HPB) and in the ionomycin-resistant (IR) fraction. Therefore, some activated T cells are resistant toward the Ca2+ ionophore. CD69 molecules are markers of the very early stage of T cell activation. However, CD4+CD69+ T cells have never been found in the IR fraction. Thus, the majority of CD4+ T lymphocytes at the early stage of activation are ionomycin-sensitive cells. The proportion of CD4+CD25+ T cells did not differ significantly in HPB and in the IR fraction. The presence of CD4+CD25+ T lymphocytes in the IR fraction reflects changes in the Ca2+-signaling pathway at this differentiation step of activated cells. Depending on the expression level of CD25 molecules, the population of CD4+CD25+ cells is divided in T-regulatory (CD25high) and proliferating (CD25low) subpopulations. The action of ionomycin results in a decrease in the portion of the CD4+CD25low T-cells, but it leads to an increase in the proportion of the CD4+CD25high T lymphocytes. Consequently, greater portion of CD4+CD25high T lymphocytes and smaller portion of CD4+CD25low T cells are IR cells. Expression of HLA-DR molecules can be used as the marker for the late activation step. The IR fraction is significantly rich in CD4+HLA DR+ T lymphocytes in comparison to the blood of the same donor. The link between different differentiation steps of CD4+ T-lymphocytes and alterations in calcium ion homeostasis is discussed. PMID- 16038613 TI - Extracellular oxidases of the lignin-degrading fungus Panus tigrinus. AB - Two extracellular oxidases (laccases) were isolated from the extracellular fluid of the fungus Panus (Lentinus) tigrinus cultivated in low-nitrogen medium supplemented with birch sawdust. The enzymes were purified by successive chromatography on columns with TEAE-cellulose and DEAE-Toyopearl 650M. Both oxidases catalyze oxidation of pyrocatechol and ABTS. Moreover, oxidase 1 also catalyzes oxidation of guaiacol, o-phenylenediamine, and syringaldazine. The enzymes have identical pH (7.0) and temperature (60-65 degrees C) optimums. Absorption spectra of the oxidases differ from the spectra of typical "blue" laccases and are similar to the spectrum of yellow oxidase. PMID- 16038614 TI - A low molecular weight isoform of hyaluronidase: purification from Indian cobra (Naja naja) venom and partial characterization. AB - A low molecular weight isoform of hyaluronidase (NNH2) has been isolated from Indian cobra (Naja naja) venom by successive chromatography on Sephadex G-75 and CM-Sephadex C-25 columns. The apparent molecular weight determined by SDS-PAGE is 52 kD, and the pI value is 9.7. NNH2 is an endoglycosidase and exhibits in vitro absolute specificity for hyaluronan; it also hydrolyzed hyaluronan in human skin sections. NNH2 is nontoxic, but it indirectly potentiates the hemorrhagic activity of hemorrhagic complex-I. Curcumin, indomethacin, and tannic acid inhibited dose dependently the degradation of hyaluronan by NNH2. PMID- 16038615 TI - PfPDE1, a novel cGMP-specific phosphodiesterase from the human malaria parasite Plasmodium falciparum. AB - This is the first report of molecular characterization of a novel cyclic nucleotide PDE (phosphodiesterase), isolated from the human malaria parasite Plasmodium falciparum and designated PfPDE1. PfPDE1 cDNA encodes an 884-amino acid protein, including six putative transmembrane domains in the N-terminus followed by a catalytic domain. The PfPDE1 gene is a single-copy gene consisting of two exons and a 170 bp intron. PfPDE1 transcripts were abundant in the ring form of the asexual blood stages of the parasite. The C-terminal catalytic domain of PfPDE1, produced in Escherichia coli, specifically hydrolysed cGMP with a K(m) value of 0.65 microM. Among the PDE inhibitors tested, a PDE5 inhibitor, zaprinast, was the most effective, having an IC50 value of 3.8 microM. The non specific PDE inhibitors IBMX (3-isobutyl-1-methylxanthine), theophylline and the antimalarial chloroquine had IC50 values of over 100 microM. Membrane fractions prepared from P. falciparum at mixed asexual blood stages showed potent cGMP hydrolytic activity compared with cytosolic fractions. This hydrolytic activity was sensitive to zaprinast with an IC50 value of 4.1 microM, but insensitive to IBMX and theophylline. Furthermore, an in vitro antimalarial activity assay demonstrated that zaprinast inhibited the growth of the asexual blood parasites, with an ED50 value of 35 microM. The impact of cyclic nucleotide signalling on the cellular development of this parasite has previously been discussed. Thus this enzyme is suggested to be a novel potential target for the treatment of the disease malaria. PMID- 16038616 TI - Varying the heart rate response to dynamic exercise in pacemaker-dependent subjects: effects on cardiac output and cerebral blood velocity. AB - Cerebral blood flow increases upon the transition from rest to moderate exercise, but becomes affected when the ability to raise CO (cardiac output) is limited. HR (heart rate) is considered to contribute significantly to the increase in CO in the early stages of dynamic exercise. The aim of the present study was to test whether manipulation of the HR response in patients dependent on permanent rate responsive ventricular pacing contributes to the increase in CO, MCA V(mean) [mean MCA (middle cerebral artery) velocity] and work capacity during exercise. The effect of setting the pacemaker to DSS ('default' sensor setting) compared with OSS ('optimized' sensor setting) on blood pressure, CO, SV (stroke volume) and MCA V(mean) was evaluated during ergometry cycling. From rest to exercise at 75 W, the rise in HR in OSS [from 73 (65-87) to 116 (73-152) beats/min; P<0.05] compared with DSS [70 (60-76) to 97 (67-117) beats/min; P<0.05] was larger. There was an increase in SV during exercise with DSS, but not with OSS, such that, at all workloads, SVs were greater during DSS than OSS. The slope of the HR-CO relationship was larger with DSS than OSS (P<0.05). From rest to exercise, MCA V(sys) (systolic MCA velocity) increased in OSS and DSS, and MCA V(dias) (diastolic MCA velocity) was reduced with DSS. No changes were observed in MCA V(mean). Manipulation of the pacemaker setting had no effect on the maximal workload [133 (100-225) W in OSS compared with 129 (75-200) W in DSS]. The results indicate that, in pacemaker-dependent subjects with complete heart block and preserved myocardial function, enhancing the HR response to exercise neither augments CO by a proportional offset of the exercise-induced increase in SV nor improves cerebral perfusion. PMID- 16038617 TI - A large displacement of the SXN motif of Cys115-modified penicillin-binding protein 5 from Escherichia coli. AB - Penicillin-binding proteins (PBPs), which are the lethal targets of beta-lactam antibiotics, catalyse the final stages of peptidoglycan biosynthesis of the bacterial cell wall. PBP 5 of Escherichia coli is a D-alanine CPase (carboxypeptidase) that has served as a useful model to elucidate the catalytic mechanism of low-molecular-mass PBPs. Previous studies have shown that modification of Cys115 with a variety of reagents results in a loss of CPase activity and a large decrease in the rate of deacylation of the penicilloyl-PBP 5 complex [Tamura, Imae and Strominger (1976) J. Biol. Chem. 251, 414-423; Curtis and Strominger (1978) J. Biol. Chem. 253, 2584-2588]. The crystal structure of wild-type PBP 5 in which Cys115 fortuitously had formed a covalent adduct with 2 mercaptoethanol was solved at 2.0 A (0.2 nm) resolution, and these results provide a structural rationale for how thiol-directed reagents lower the rate of deacylation. When compared with the structure of the unmodified wild-type enzyme, a major change in the architecture of the active site is observed. The two largest differences are the disordering of a loop comprising residues 74-90 and a shift in residues 106-111, which results in the displacement of Ser110 of the SXN active-site motif. These results support the developing hypothesis that the SXN motif of PBP 5, and especially Ser110, is intimately involved in the catalytic mechanism of deacylation. PMID- 16038618 TI - Conformation-dependent inactivation of human betaine-homocysteine S methyltransferase by hydrogen peroxide in vitro. AB - Betaine-homocysteine S-methyltransferase (BHMT) transfers a methyl group from betaine to Hcy to form DMG (dimethylglycine) and Met. The reaction is ordered Bi Bi; Hcy is the first substrate to bind and Met is the last product off. Using intrinsic tryptophan fluorescence [Castro, Gratson, Evans, Jiracek, Collinsova, Ludwig and Garrow (2004) Biochemistry 43, 5341-5351], it was shown that BHMT exists in three steady-state conformations: enzyme alone, enzyme plus occupancy at the first substrate-binding site (Hcy or Met), or enzyme plus occupancy at both substrate-binding sites (Hcy plus betaine, or Hcy plus DMG). Betaine or DMG alone do not bind to the enzyme, indicating that the conformational change associated with Hcy binding creates the betaine-binding site. CBHcy [S-(d carboxybutyl)-D,L-homocysteine] is a bisubstrate analogue that causes BHMT to adopt the same conformation as the ternary complexes. We report that BHMT is susceptible to conformation-dependent oxidative inactivation. Two oxidants, MMTS (methyl methanethiosulphonate) and hydrogen peroxide (H2O2), cause a loss of the enzyme's catalytic Zn (Zn2+ ion) and a correlative loss of activity. Addition of 2-mercaptoethanol and exogenous Zn after MMTS treatment restores activity, but oxidation due to H2O2 is irreversible. CD and glutaraldehyde cross-linking indicate that H2O2 treatment causes small perturbations in secondary structure but no change in quaternary structure. Oxidation is attenuated when both binding sites are occupied by CBHcy, but Met alone has no effect. Partial digestion of ligand-free BHMT with trypsin produces two large peptides, excising a seven residue peptide within loop L2. CBHcy but not Met binding slows down proteolysis by trypsin. These findings suggest that L2 is involved in the conformational change associated with occupancy at the betaine-binding site and that this conformational change and/or occupancy at both ligand-binding sites protect the enzyme from oxidative inactivation. PMID- 16038619 TI - A polycystin multiprotein complex constitutes a cholesterol-containing signalling microdomain in human kidney epithelia. AB - Polycystins are plasma membrane proteins that are expressed in kidney epithelial cells and associated with the progression of ADPKD (autosomal dominant polycystic kidney disease). A polycystin multiprotein complex, including adherens junction proteins, is thought to play an important role in cell polarity and differentiation. Sucrose gradient analyses and immunoprecipitation studies of primary human kidney epithelial cells showed the polycystins and their associated proteins E-cadherin and beta-catenin distributed in a complex with the raft marker flotillin-2, but not caveolin-1, in high-density gradient fractions. The integrity of the polycystin multiprotein complex was sensitive to cholesterol depletion, as shown by cyclodextrin treatment of immunoprecipitated complexes. The overexpressed C-terminus of polycystin-1 retained the ability to associate with flotillin-2. Flotillin-2 was found to contain CRAC (cholesterol recognition/interaction amino acid) cholesterol-binding domains and to promote plasma membrane cholesterol recruitment. Based on co-association of signalling molecules, such as Src kinases and phosphatases, we propose that the polycystin multiprotein complex is embedded in a cholesterol-containing signalling microdomain specified by flotillin-2, which is distinct from classical light buoyant-density, detergent-resistant domains. PMID- 16038620 TI - Prader-Willi syndrome: clinical genetics, cytogenetics and molecular biology. AB - Prader-Willi syndrome (PWS) is a neurodevelopmental disorder that arises from lack of expression of paternally inherited genes known to be imprinted and located in the chromosome 15q11-q13 region. PWS is considered the most common syndromal cause of life-threatening obesity and is estimated at 1 in 10,000 to 20,000 individuals. A de novo paternally derived chromosome 15q11-q13 deletion is the cause of PWS in about 70% of cases, and maternal disomy 15 accounts for about 25% of cases. The remaining cases of PWS result either from genomic imprinting defects (microdeletions or epimutations) of the imprinting centre in the 15q11 q13 region or from chromosome 15 translocations. Here, we describe the clinical presentation of PWS, review the current understanding of causative cytogenetic and molecular genetic mechanisms, and discuss future directions for research. PMID- 16038621 TI - Molecular basis of ataxia telangiectasia and related diseases. AB - Ataxia telangiectasia (AT) is a rare human disease characterized by extreme cellular sensitivity to radiation and a predisposition to cancer, with a hallmark of onset in early childhood. Several human diseases also share similar symptoms with AT albeit with different degrees of severity and different associated disorders. While all AT patients contain mutations in the AT-mutated gene (ATM), most other AT-like disorders are defective in genes encoding an MRN protein complex consisting of Mre11, Rad50 and Nbs1. Both ATM and MRN function as cellular sensors to DNA double-strand breaks, which lead to the recruitment and phosphorylation of an array of substrate proteins involved in DNA repair, apoptosis and cell-cycle checkpoints, as well as gene regulation, translation initiation and telomere maintenance. ATM is a member of the family of phosphatidylinositol 3-kinase-like protein kinases (PIKK), and the discovery of many ATM substrates provides the underlying mechanisms of heterologous symptoms among AT patients. This review article focuses on recent findings related to the initial recognition of double-strand breaks by ATM and MRN, as well as a DNA dependent protein kinase complex consisting of the heterodimer Ku70/Ku80 and its catalytic subunit DNA-PKcs, another member of PIKK. This possible interaction suggests that a much greater complex is involved in sensing, transducing and co ordinating cellular events in response to genome instability. PMID- 16038622 TI - Oxidative stress and stress signaling: menace of diabetic cardiomyopathy. AB - Cardiovascular disease is the most common cause of death in the diabetic population and is currently one of the leading causes of death in the United States and other industrialized countries. The health care expenses associated with cardiovascular disease are staggering, reaching more than 350 billion dollars in 2003. The risk factors for cardiovascular disease include high fat/cholesterol levels, alcoholism, smoking, genetics, environmental factors and hypertension, which are commonly used to gauge an individual's risk of cardiovascular disease and to track their progress during therapy. Most recently, these factors have become important in the early prevention of cardiovascular diseases. Oxidative stress, the imbalance between reactive oxygen species production and breakdown by endogenous antioxidants, has been implicated in the onset and progression of cardiovascular diseases such as congestive heart failure and diabetes-associated heart dysfunction (diabetic cardiomyopathy). Antioxidant therapy has shown promise in preventing the development of diabetic heart complications. This review focuses on recent advances in oxidative stress theory and antioxidant therapy in diabetic cardiomyopathy, with an emphasis on the stress signaling pathways hypothesized to be involved. Many of these stress signaling pathways lead to activation of reactive oxygen species, major players in the development and progression of diabetic cardiomyopathy. PMID- 16038623 TI - Ion channelopathy and hyperphosphorylation contributing to cardiac arrhythmias. AB - The occurrence of cardiac arrhythmias is related to the abnormality of ion channels not only in sarcolemma but also in the sarcoplasmic reticulum, which regulates the process of calcium release and up-take intracellularly. Patterns of ion channelopathy in the sarcolemma can be divided into single channel disorder from gene mutations and multiple channels disorder in a diseased hypertrophied heart. Abnormal RyR2, FKBP12.6, SERCA2a, and PLB are also involved in the initiation of cardiac arrhythmias. Maladjustment by hyperphosphorylation on the ion channels in the sarcolemma and RyR2-FKBP12.6 and SERCA2a-PLB is discussed. Hyperphosphorylation, which is the main abnormality upstream to ion channels, can be targeted for suppressing the deterioration of ion channelopathy in terms of new drug discovery in the treatment and prevention of malignant cardiac arrhythmias. PMID- 16038624 TI - Is COX-2 a perpetrator or a protector? Selective COX-2 inhibitors remain controversial. AB - COX-2(cyclooxygenase-2) has sparked a surge in pharmaceutical interest since its discovery at the beginning of the 1990s. Several COX-2 selective inhibitors that avoid gastrointestinal side effects have been successfully launched into the market in recent years. The first selective COX-2 inhibitor, celecoxib, entered the market in December 1998 [corrected] However, there are a few organs that physiologically and functionally express COX-2, particularly the glomeruli of the kidney and the cortex of the brain. Inhibition of COX-2 expression in these organs possibly causes heart attack and stroke in long-term COX-2 inhibitor users. Recently, a USA Food and Drug Agency (FDA) advisory panel re-evaluated COX 2 inhibitors and unanimously concluded that the entire class of COX-2 inhibitors increase the risk of cardiovascular problems. Thus the use of COX-2 inhibitors is still controversial, and there is a challenge for not only pharmacologists, but also the pharmaceutical industry, to develop improved painkilling and anti inflammatory drugs. This may involve exploring a new generation of COX-2 inhibitors with different inhibitory mechanisms through computer-aided design, screening different sources of inhibitors with lower selectivity, or seeking completely new targets. Synthetic COX-2 inhibitors have high selectivity and the advantage of irreversible inhibition, whereas naturally derived COX-2 inhibitors have lower selectivity and fewer side effects, with the medical effects in general not being as striking as those achieved using synthetic inhibitors. This review discusses the mechanism of COX-2 inhibitor therapy and a possible new way of exploration in the development of anti-inflammatory, analgetic, and antipyretic drugs. PMID- 16038625 TI - Interleukin-1 beta induction of neuron apoptosis depends on p38 mitogen-activated protein kinase activity after spinal cord injury. AB - AIM: Interleukin-1 beta (IL-1beta) has been implicated as an extracellular signal in the initiation of apoptosis in neurons and oligodendrocytes after spinal cord injury (SCI). To further characterize the apoptotic cascade initiated by IL-1beta after SCI, we examined the expression of IL-1beta, p38 mitogen-activated protein kinase (p38 MAPK) and caspase-3 after SCI, and further investigated whether p38 MAPK was involved in neuron apoptosis induced by IL-1beta. METHODS: Adult rats were given contusion SCI at the T-10 vertebrae level with a weight-drop impactor (10 g weight dropped 25.0 mm). The expression levels of IL-1beta, p38 MAPK and caspase-3 after SCI were assessed with Western blots, immunohistochemistry staining, and real time reverse transcription polymerase chain reactions (RT PCR). Neuron apoptosis was assessed with the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) method. RESULTS: Increased levels of IL-1beta and p38 MAPK were observed soon after injury, with a peak in expression levels within 6 h of injury. By 24 h after injury, caspase-3 expression was markedly increased in the injured spinal cord. TUNEL-positive cells were first observed in the lesioned area 6 h after SCI. The largest number of TUNEL-positive cells was observed at 24 h post-SCI. Intrathecal injection of the IL-1 receptor antagonist IL-1Ra significantly reduced expression of p38 MAPK and caspase-3, and reduced the number of TUNEL positive cells. Moreover, intrathecal injection of an inhibitor of p38 MAPK, SB203580, also significantly reduced the expression of caspase-3, and reduced the number of TUNEL-positive cells in the injured spinal cord. CONCLUSION: The p38MAPK signaling pathway plays an important role in IL-1beta mediated induction of neuron apoptosis following SCI in rats. PMID- 16038626 TI - Sodium ferulate prevents amyloid-beta-induced neurotoxicity through suppression of p38 MAPK and upregulation of ERK-1/2 and Akt/protein kinase B in rat hippocampus. AB - AIM: To observe whether an amyloid beta (Abeta)-induced increase in interleukin (IL)-1beta was accompanied by an increase in the p38 mitogen-activated protein kinase (MAPK) pathway and a decrease in the cell survival pathway, and whether sodium ferulate (SF) treatment was effective in preventing these Abeta-induced changes. METHODS: Rats were injected intracerebroventricularly with Abeta25-35. Seven days after injection, immunohistochemical techniques for glial fibrillary acidic protein (GFAP) were used to determine the astrocyte infiltration and activation in hippocampal CA1 areas. The expression of IL-1beta, extracellular signal-regulated kinase (ERK), p38 MAPK, Akt/protein kinase B (PKB), Fas ligand and caspase-3 were determined by Western blotting. The caspase-3 activity was measured by cleavage of the caspase-3 substrate (Ac-DEVD-pNA). Reverse transcription-polymerase chain reaction was used to analyze the changes in IL 1beta mRNA levels. RESULTS: Intracerebroventricular injection of Abeta25-35 elicited astrocyte activation and infiltration and caused a strong inflammatory reaction characterized by increased IL-1beta production and elevated levels of IL 1beta mRNA. Increased IL-1beta synthesis was accompanied by increased activation of p38 MAPK and downregulation of phospho-ERK and phospho-Akt/PKB in hippocampal CA regions prepared from Abeta-treated rats, leading to cell death as assessed by activation of caspase-3. SF significantly prevented Abeta-induced increases in IL 1beta and p38 MAPK activation and also Abeta-induced changes in phospho-ERK and phospho-Akt/PKB expression levels. CONCLUSION: SF prevents Abeta-induced neurotoxicity through suppression of p38 MAPK activation and upregulation of phospho-ERK and phospho-Akt/PKB expression. PMID- 16038627 TI - Neuroprotective effects of cyclooxygenase-2 inhibitor celecoxib against toxicity of LPS-stimulated macrophages toward motor neurons. AB - AIM: To establish an in vitro injured motor neuronal model and investigate the neuroprotective effects and possible mechanism of celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, on this model. METHODS: After macrophages were stimulated with lipopolysaccharide (LPS)+interferon-gamma (IFN-gamma) in the presence or absence of celecoxib for 24 h, the cell-free supernatant of LPS stimulated macrophages was transferred to the culture of NSC34 cells. Viability of NSC34 cells was assessed by MTT assay after a further 24 h and 72 h incubation. After macrophages were stimulated by LPS+IFN-gamma for 12 h or 24 h, the release of prostaglandin E2 (PGE2), nitric oxide (NO), reactive oxygen species (ROS), tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL 1beta) from macrophages was measured by radioimmunoassay, Griess assay, fluorescence assay and enzyme-linked immunosorbent assay, respectively. The mRNA levels of COX-2, inducible nitric oxide synthase (iNOS), TNF-alpha and IL-1beta in macrophages were determined by reverse transcription-polymerase chain reaction after macrophages were stimulated for 6 h and 12 h. RESULTS: The supernatant of LPS-stimulated mouse macrophages induced the death of NSC34 cells and celecoxib protected the NSC34 cells against this toxicity. The LPS-induced increases in the release of PGE2, NO, TNF-alpha and IL-1beta from macrophages were attenuated by pre-treatment with celecoxib. However, celecoxib showed no effect on the ROS levels upregulated by LPS+IFN-gamma in the macrophage supernatant. The mRNA levels of COX-2, iNOS, TNF-alpha and IL-1beta were increased in LPS-activated macrophages and, except COX-2, reduced by pre-treatment with celecoxib. CONCLUSION: An in vitro injured motor neuronal model was established by using the toxicity of LPS-stimulated mouse macrophages toward motor neuronal NSC34 cells. In this model, celecoxib exerted neuroprotective effects on motor neurons via an inhibition of the neurotoxic secretions from activated macrophages. PMID- 16038628 TI - Blood pressure variability and baroreflex sensitivity are not different in spontaneously hypertensive rats and stroke-prone spontaneously hypertensive rats. AB - AIM: To demonstrate and compare hemodynamic phenotypes of blood pressure (BP), blood pressure variability (BPV) and baroreflex sensitivity (BRS) in genetic hypertensive rats. METHODS: BP was recorded continuously in conscious, freely moving rats using a computerized technique. BPV was expressed as the standard deviation of beat-to-beat BP values during a 1-h period. BRS was determined by measuring the heart period prolongation in response to the elevation in BP produced by an intravenous injection of phenylephrine. RESULTS: Body weight and heart period were not different between spontaneously hypertensive rats (SHR) and stroke-prone spontaneously hypertensive rats (SHR-SP) at the age of 15 weeks. The BP level was markedly higher in SHR-SP than SHR, whereas there were no significant differences in BPV and BRS. Quantitatively, systolic, diastolic and mean BP were significantly elevated by 36.9%, 42.9% and 39.5%, respectively, in SHR-SP compared with SHR (P < 0.01). However, their variabilities were elevated only by 14.0%, 0.4% and 10.1%, respectively, without statistical significance (P > 0.05). CONCLUSION: BPV and BRS were not changed in parallel with the BP alterations in SHR and SHR-SP. PMID- 16038629 TI - Adeno-associated virus-mediated bone morphogenetic protein-7 gene transfer induces C2C12 cell differentiation into osteoblast lineage cells. AB - AIM: To investigate the effects of bone morphogenetic protein-7 (BMP7)-expressing recombinant adeno-associated virus (AAV) vector on the differentiation of C2C12 cells. METHODS: AAV-BMP7 was packaged by infecting the stable cell clone BHK-21 (integrated with recombinant AAV vector plasmid pSNAV-BMP7) with recombinant herpes simplex virus type 1, which expresses AAV-2 Rep and Cap and possesses AAV packaging functions. Following infection with AAV-BMP7 at multiplicities of infection of 1 x 10(5) vector genomes per cell and subsequent culture, C2C12 cells were assessed qualitatively for BMP7 production, alkaline phosphatase activity, osteocalcin production and Cbfal and MyoD expression. RESULTS: C2C12 cells transduced with AAV-BMP7 could produce BMP7 protein until d 28. Alkaline phosphatase in the cultured C2C12 cell lysate was elevated. Secreted osteocalcin in the culture medium was detectable at d 12 and Cbfal mRNA expression level was upregulated, coinciding with downregulation of MyoD in a temporal manner. CONCLUSION: The present in vitro study demonstrated that AAV-BMP7 could infect and efficiently convert C2C12 cells from myoblasts into osteoblast lineage cells. PMID- 16038630 TI - Apoptosis initiated by carbon tetrachloride in mitochondria of rat primary cultured hepatocytes. AB - AIM: To investigate the mitochondria-initiated apoptosis pathway involved in Carbon tetrachloride (CCl4) hepatotoxicity in vitro. METHODS: Several cytotoxicity endpoints, including WST-8 metabolism, lactate dehydrogenase leakage and morphological changes, were examined. The 5,5'-dithio-bis(2-nitrobenzoic acid) reaction was used to measure reduced glutathione level, and the malondialdehyde level was determined using the thiobarbituric acid assay. The release of cytochrome c and Bcl-X(L) was detected by Western blot. Caspase-3 activity was measured using the fluorogenic substrate Ac-DEVD-AMC. DNA fragmentation was used to evaluate cell apoptosis. RESULTS: A time- and dose dependent decrease in cellular glutathione content was observed, along with a concomitant increase in malondialdehyde levels following the application of CCl4. Caspase 3 activity was stimulated at all doses of CCl4, with the most significant activation at 3 mmol/L. Cytochrome c was released obviously after CCl4 treatment. A time-dependent decrease in Bcl-X(L) expression was observed. DNA fragmentation results revealed apoptosis and necrosis following CCl4 treatment. CONCLUSION: Oxidative damage is one of the essential mechanisms of CCl4 hepatotoxicity, which triggers apoptosis via the mitochondria-initiated pathway. PMID- 16038631 TI - Expression of feeding-related peptide receptors mRNA in GT1-7 cell line and roles of leptin and orexins in control of GnRH secretion. AB - AIM: To investigate the expression of feeding-related peptide receptors mRNA in GT1-7 cell line and roles of leptin and orexins in the control of GnRH secretion. METHODS: Receptors of bombesin3, cholecystokinin (CCK)-A, CCK-B, glucagon-like peptide (GLP)1, melanin-concentrating hormone (MCH)1, orexin1, orexin2, neuromedin-B, neuropeptide Y (NPY)1 and NPY5, neurotensin (NT)1, NT2, NT3, and leptin receptor long form mRNA in GT1-7 cells were detected by reversed transcriptase-polymerase chain reaction. GT1-7 cells were treated with leptin, orexin A and orexin B at a cohort of concentrations for different lengths of time, and GnRH in medium was determined by radioimmunoassay (RIA). RESULTS: Receptors of bombesin 3, CCK-B, GLP1, MCH1, orexin1, neuromedin-B, NPY1, NPY5, NT1, NT3, and leptin receptor long form mRNA were expressed in GT1-7 cells, of which, receptors of GLP1, neuromedin-B, NPY1, and NT3 were highly expressed. No amplified fragments of orexin2, NT2, and CCK-A receptor cDNA were generated with GT1-7 RNA, indicating that the GT1-7 cells did not express mRNA of them. Leptin induced a significant stimulation of GnRH release, the results being most significant at 0.1 nmol/L for 15 min. In contrast to other studies in hypothalamic explants, neither orexin A nor orexin B affected basal GnRH secretion over a wide range of concentrations ranging from 1 nmol/L to 500 nmol/Lat 15, 30, and 60 min. CONCLUSION: Feeding and reproductive function are closely linked. Many orexigenic and anorexigenic signals may control feeding behavior as well as alter GnRH secretion through their receptors on GnRH neurons. PMID- 16038632 TI - Puerarin reduces increased c-fos, c-jun, and type IV collagen expression caused by high glucose in glomerular mesangial cells. AB - AIM: Increased expression of c-fos, c-jun and type IV collagen (CoIV) in glomerular mesangial cells (GMC) are important characteristics of diabetic nephropathy. Both c-fos and c-jun regulate the gene expression of extracellular matrix components, and CoIV is the main component of the extracellular matrix. It has been reported that puerarin inhibits aggregation of the extracellular matrix in diabetic rats by an as yet unknown mechanism. The aim of this study is to investigate the effect of puerarin on c-fos, c-jun and CoIV expression in GMC cultured in medium containing 5.6 or 27.8 mmol/L glucose. METHODS: The expressions of c-fos and c-jun were measured at the protein level using flow cytometry. CoIV content was detected using radioimmunoassay. Protein kinase C (PKC) activity was measured using liquid scintillation counting. RESULTS: Puerarin (10(-5) mmol/L) significantly ameliorated the high-glucose effect on c fos, c-jun and CoIV expression. This effect is accompanied by a reduced PKC activity in these cells. CONCLUSION: Our results suggest that reduced PKC activity and expression of c-fos and c-jun in GMC might participate in the mechanisms underlying the therapeutic effect of puerarin on diabetic nephropathy. PMID- 16038633 TI - Protective effect of membrane cofactor protein against complement-dependent injury. AB - AIM: To evaluate the protective role of membrane cofactor protein (MCP, CD46) on complement-dependent injury. METHODS: MCP was separated by ion exchange chromatography on a DEAE sephadex A-50 column from pig erythrocyte ghosts. Its protective effect was tested in models such as cobra venom factor (CVF)-induced platelet metamorphosis and aggregation, human serum-induced injury in isolated working guinea pig heart and reverse passive Arthus reaction. RESULTS: MCP inhibited CVF-induced platelet metamorphosis with an IC50 of 56.7 mg/L+/-2.6 mg/L, and prevented injury induced by activated complement in isolated working guinea pig hearts. In the rat model of reverse Arthus reaction, MCP relieved the skin lesions induced by immune complexes. CONCLUSION: MCP has a protective effect against complement-dependent injury. PMID- 16038634 TI - Protective effects of L-arginine on pulmonary oxidative stress and antioxidant defenses during exhaustive exercise in rats. AB - AIM: To assess the effects of L-arginine (L-Arg) supplementation on pulmonary oxidative stress and antioxidant defenses in rats after exhaustive exercise. METHODS: Rats were randomly divided into four groups: sedentary control (SC), sedentary control with L-Arg treatment (SC+Arg), exhaustive exercise with control diet (E) and exhaustive exercise with L-Arg treatment (E+Arg). Rats in groups SC+Arg and E+Arg received a 2% L-Arg diet. Rats in groups E and E+Arg underwent an exhaustive running test on a motorized treadmill. Pulmonary oxidative stress indices [xanthine oxidase (XO), myeloperoxidase (MPO), and malondialdehyde (MDA)] and antioxidant defense systems [superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GR), and glutathione (GSH)] were investigated in this study. RESULTS: L-Arg supplementation significantly reduced exercise-induced elevations of XO and MPO activities in lung. L-Arg reversed the exercise-induced increase in SOD and GR activities, but increased CAT and GPX activities. L-Arg administration also significantly increased the GSH levels in plasma. CONCLUSION: L-Arg supplementation can prevent elevations of XO and MPO activities in the lung and favorably influence pulmonary antioxidant defense systems after exhaustive exercise. PMID- 16038635 TI - Cytotoxicity, apoptosis induction, and mitotic arrest by a novel podophyllotoxin glucoside, 4DPG, in tumor cells. AB - AIM: To define the in vitro cytotoxic activities of 4-demethyl picropodophyllotoxin 7'-O-beta-D-glucopyranoside (4DPG), a new podophyllotoxin glucoside. METHODS: Antiproliferation activity was measured in several tumor cell lines by using the microculture tetrazolium MTT assays. Cell cycle distribution was analyzed using flow cytometry and mitosis index assays. Furthermore, transmission electron microscopy, TUNEL, DNA agarose electrophoresis, and activated caspase-3 were used to analyze the induction of apoptotic cell death. Moreover, intracellular changes in the cytoskeleton were detected using immunocytochemistry. RESULTS: 4DPG effectively inhibited the proliferation of cancer cells (HeLa, CNE, SH-SY5Y, and K562 cell lines). For the K562 cell line, the antiproliferation effect of 4DPG was much more potent than that of etoposide (IC50 value: 7.79 x 10(-9) mol/L for 4DPG vs 2.23 x 10(-5) mol/L for etoposide). Further, 4DPG blocked the cell cycle in the mitotic phase. The induction of apoptosis and elevated levels of activated caspase-3 were confirmed in cells treated with 4DPG. The microtubule skeleton of HeLa cells was disrupted immediately after treatment with 4DPG. CONCLUSION: The cytotoxicity of 4DPG is due to its inhibition of the microtubule assembly of cancer cells at a low concentration, thus inducing apoptosis. These properties qualify 4DPG to be a potential antitumor drug. PMID- 16038636 TI - Effect of curcumin on multidrug resistance in resistant human gastric carcinoma cell line SGC7901/VCR. AB - AIM: To investigate the reversal effects of curcumin on multidrug resistance (MDR) in a resistant human gastric carcinoma cell line. METHODS: The cytotoxic effect of vincristine (VCR) was evaluated by MTT assay. The cell apoptosis induced by VCR was determined by propidium iodide (PI)-stained flow cytometry (FCM) and a morphological assay using acridine orange (AO)/ethidium bromide (EB) dual staining. P-glycoprotein (P-gp) function was demonstrated by the accumulation and efflux of rhodamine123 (Rh123) using FCM. The expression of P-gp and the activation of caspase-3 were measured by FCM using fluorescein isothiocyanate (FITC)-conjugated anti-P-gp and anti-cleaved caspase-3 antibodies, respectively. RESULTS: Curcumin, at concentrations of 5 micromol/L, 10 micromol/L, or 20 micromol/L, had no cytotoxic effect on a parent human gastric carcinoma cell line (SGC7901) or its VCR-resistant variant cell line (SGC7901/VCR). The VCR-IC50 value of the SGC7901/VCR cells was 45 times more than that of the SGC7901cells and the SGC7901/VCR cells showed apoptotic resistance to VCR. SGC7901/VCR cells treated with 5 micromol/L, 10 micromol/L, or 20 micromol/L curcumin decreased the IC50 value of VCR and promoted VCR-mediated apoptosis in a dose-dependent manner. Curcumin (10 micromol/L) increased Rh123 accumulation and inhibited the efflux of Rh123 in SGC7901/VCR cells, but did not change the accumulation and efflux of Rh123 in SGC7901 cells. P-gp was overexpressed in SGC7901/VCR cells, whereas it was downregulated after a 24-h treatment with curcumin (10 micromol/L). Resistant cells treated with 1 mumol/L VCR alone showed 77% lower levels of caspase-3 activation relative to SGC7901 cells, but the activation of caspase-3 in the resistant cell line increased by 44% when cells were treated with VCR in combination with curcumin. CONCLUSION: Curcumin can reverse the MDR of the human gastric carcinoma SGC7901/VCR cell line. This might be associated with decreased P-gp function and expression, and the promotion of caspase-3 activation in MDR cells. PMID- 16038637 TI - Effects of navelbine and docetaxel on gene expression in lung cancer cell strains. AB - AIM: To search genes sensitivity to the anti-cancer drugs navelbine (NVB) and docetaxel (DOC) in small-cell lung cancer (SCLC) and non-small-cell lung cancer (NSCLC) cell strains. METHODS: The sensitivity of 4 strains of SCLC and 6 strains of NSCLC to NVB and DOC was evaluated using the MTT assay. The expression of 1291 sensitive-related genes to the anti-cancer drugs in 10 lung cancer cell strains was measured using cDNA macroarrays and the relationship was analyzed. RESULTS: In total, there were 56 (r > or = 0.4) genes sensitive to NVB and DOC. For NVB: 36 genes were sensitive to NVB, 20 co-expressed genes between the SCLC+NSCLC set and the NSCLC set; 27 expressed genes and 7 specially expressed genes in the SCLC+NSCLC set; and 29 expressed genes and 9 specially expressed genes in the NSCLC set. For DOC, 50 genes were sensitive to DOC, 12 co-expressed genes between the SCLC+NSCLC set and the NSCLC set; 24 expressed genes and 12 specially expressed genes in the SCLC+NSCLC set; and 38 expressed genes and 26 specially expressed genes in the NSCLC set. The genes sensitive to NVB and DOC in lung cancer cell stains were mainly divided into the following 4 categories: signal transduction molecules, cell factors, transcription factors and metabolism related enzymes and inhibitors. CONCLUSIONS: There were obvious differences in genes related to NVB and DOC between SCLC and NSCLC cell strains, but the same as categories of function. PMID- 16038638 TI - Competing outcomes. PMID- 16038639 TI - Colon cancer clinical practice guidelines in oncology. PMID- 16038640 TI - Rectal cancer clinical practice guidelines in oncology. PMID- 16038641 TI - Anal canal cancer clinical practice guidelines in oncology. PMID- 16038642 TI - Surgical management of colorectal cancer in the laparoscopic era: a review of prospective randomized trials. AB - The benefits of laparoscopy in benign diseases are quite clear. Patients generally can expect smaller incisions, less narcotic usage, quicker return of bowel function, and shorter hospitalizations. The benefits of laparoscopy in oncologic surgery are less clear, and laparoscopic oncology surgery has many critics. Early reports of long surgical times, high operating room costs, and alarming rates of port-site recurrences after laparoscopic colectomy for colorectal cancer all but stopped this less-invasive approach outside the confines of clinical protocols. As the results of larger retrospective studies began to refute these earlier detrimental claims, prospective randomized trials began to take a foothold. In this article, we review these randomized trials with particular attention to the perioperative effects of laparoscopic colectomy and the short-term oncologic outcomes. PMID- 16038643 TI - Chemotherapy for metastatic colorectal cancer. AB - The past decade has seen a significant survival improvement for patients with metastatic colorectal cancer, fueled in large part by the arrival of active novel chemotherapeutic drugs and their incorporation into combination regimens. Several randomized trials have successfully integrated oxaliplatin and irinotecan into previously existing 5-fluorouracil (5-FU)-based regimens for advanced colorectal cancer, resulting in median survivals that have risen from 9 months to almost 2 years. Even as the ideal combinations and sequences of these regimens are elucidated, targeted therapies such as recently approved bevacizumab and cetuximab have been added to treatment protocols, with favorable consequences. We review the evolution of primary chemotherapy for advanced colorectal cancer, focusing on the trials that have led to the new standard first-line treatments. We also review the data on newer targeted therapies, especially in combination with cytotoxic therapy. PMID- 16038644 TI - Local excision for rectal cancer. AB - Local excision can be a definitive surgical procedure for some early cancers of the rectum, and it is an appealing technique in many ways. It reduces overall surgical trauma and can assure sphincter preservation. It is also associated with low morbidity. However, this technique has a number of limitations also. This article discusses the results of local excision for rectal carcinoma, including histologic features that impact survival and local recurrence-free rates, the importance of careful patient selection, and the potential role of adjuvant and salvage therapies. Technical considerations and alternative therapies are also discussed. PMID- 16038645 TI - Myeloid growth factors clinical practice guidelines in oncology. PMID- 16038646 TI - Guidelines of the National Comprehensive Cancer Network on the use of myeloid growth factors with cancer chemotherapy: a review of the evidence. AB - The prophylactic use of myeloid growth factors reduces the risk of chemotherapy induced neutropenia and its complications, including febrile neutropenia and infection-related mortality. Perhaps most importantly, the prophylactic use of colony-stimulating factors (CSFs) has been shown to reduce the need for chemotherapy dose reductions and delays that may limit chemotherapy dose intensity, thereby increasing the potential for prolonged disease-free and overall survival in the curative setting. National surveys have shown that the majority of patients with potentially curable breast cancer or non-Hodgkin's lymphoma (NHL) do not receive prophylactic CSF support. In this issue, the National Comprehensive Cancer Network presents guidelines for the use of myeloid growth factors in patients with cancer. These guidelines recommend a balanced clinical evaluation of the potential benefits and harms associated with chemotherapy to define the treatment intention, followed by a careful assessment of the individual patient's risk for febrile neutropenia and its complications. The decision to use prophylactic CSFs is then based on the patient's risk and potential benefit from such treatment. The routine prophylactic use of CSFs in patients receiving systemic chemotherapy is recommended in patients at high risk (>20%) of developing febrile neutropenia or related complications that may compromise treatment. Where compelling clinical indications are absent, the potential for CSF prophylaxis to reduce or offset costs by preventing hospitalization for FN should be considered. The clinical, economic, and quality of life data in support of these recommendations are reviewed, and important areas of ongoing research are highlighted. PMID- 16038647 TI - Senior adult oncology clinical practice guidelines in oncology. PMID- 16038648 TI - Older patients, cognitive impairment, and cancer: an increasingly frequent triad. AB - The incidence of both cancer and cognitive impairments from various origins increases with age. Oncologists are increasingly being confronted with cancers occurring in patients with cognitive impairment, yet very few studies have addressed the problem. Cognitive impairment affects a patients' survival to an extent similar to an average cancer, and this can be an important thing to consider, especially in the adjuvant setting. Cognitive impairment also predisposes patients to delirium in the surgery setting or during hospitalization. Because effective preventive measures exist, careful attention should be paid to identifying patients at risk. Cognitive impairment does not automatically mean inability to consent, but particular precautions should be taken. For outpatient treatments such as chemotherapy, a comprehensive multidisciplinary approach is key for a good outcome. Proper caregiver support should be ensured up-front, and aggressive supportive care should be used. In the setting of an experienced geriatric oncology team, patients with cognitive impairment appear more likely to receive standard oncologic therapies. Cancer patients with cognitive impairment are at high risk of concomitant depression. PMID- 16038649 TI - Spontaneous long-term changes of corneal power and astigmatism after suture removal after penetrating keratoplasty using a regression model. AB - PURPOSE: To assess the diagnosis-based spontaneous long-term changes in corneal power and refraction with a regression model in the all-sutures-out time period following non-mechanical penetrating keratoplasty (PK). DESIGN: Retrospective non randomized clinical trial. METHODS: setting: Clinical practice. study population: 147 eyes [47 Fuchs dystrophy (FD); 100 keratoconus (KC)] were studied after suture removal in this retrospective longitudinal study. main outcome measures: Zeiss keratometry [equivalent power (KEQ) and astigmatism (KAST)], corneal topography analysis [equivalent power (TEQ) and astigmatism (TAST)], and subjective refractometry [spherical equivalent (SEQ) and refractive cylinder (RAST)] were assessed in at least three up to 16 ophthalmologic examinations in the all-sutures-out time period. observation procedure: The time course of each target variable was analyzed in a longitudinal manner (time interval > or = 12 months) separately for each patient with a linear regression model. RESULTS: Post keratoplasty follow-up ranged from 31 months to 10.3 years. In the linear regression model, the annual change in FD/KC showed an increase/a decrease in KEQ (0.29 +/- 0.50/-0.63 +/- 0.46 diopters, P = .02) and an increase/a decrease in TEQ (0.37 +/- 0.54/-0.69 +/- 0.49 diopters, P = .04) corresponding to a decrease/an increase in SEQ (-0.31 +/- 0.47/0.63 +/- 0.43 diopters, P = .02). KAST/TAST/RAST showed a minimal annual decrease (-0.06 +/- 0.41/-0.05 +/- 0.45/ 0.06 +/- 0.41 diopters) in FD but an increase in KC (0.46 +/- 0.41/0.51 +/- 0.43/0.46 +/- 0.38 diopters) (P = .05/0.06/0.12). CONCLUSIONS: In the follow-up after post-keratoplasty suture removal, patients with FD/KC tend to develop a spontaneous myopic shift (steepening of the cornea)/hyperopic shift (flattening of the cornea). In contrast with those with FD, patients with KC should be counseled on the fact that astigmatism may increase again over time after suture removal. PMID- 16038650 TI - Birdshot retinochoroidopathy: ocular complications and visual impairment. AB - PURPOSE: To describe the incidence of vision loss and of ocular complications attributable to birdshot retinochoroidopathy and to describe the association between therapy and the incidence thereof. DESIGN: Retrospective cohort study. METHODS: SETTING: Single-center, academic practice. STUDY POPULATION: Forty patients with birdshot retinochoroidopathy were evaluated from January 1984 through March 2004. OBSERVATION PROCEDURE: Demographic and clinical information on patients diagnosed with birdshot retinochoroidopathy was collected. MAIN OUTCOME MEASURES: Visual acuity and visual field loss; ocular complications including cystoid macular edema (CME). RESULTS: In affected eyes, the frequencies of vision loss to 20/50 or worse and to 20/200 or worse and of CME at presentation were 33%, 13%, and 20%, respectively. Patients who presented with a duration of disease of > or = 30 months had higher frequencies of visual impairment to 20/50 or worse (68% vs 32%; P = .004) and to 20/200 or worse (32% vs 9%; P = .01), and had a higher frequency of CME (38% vs 14%; P = .02) than patients who presented with a duration of disease <30 months. The incidence rates on follow-up for vision loss to 20/50 or worse and to 20/200 or worse were 13% and 4% per eye-year (EY), respectively. The incidence of CME was 10%/EY. Use of immunosuppressive drug therapy was associated with a reduced risk of developing CME (relative risk = 0.17; 95% confidence interval: 0.05, 0.64; P = .009). CONCLUSIONS: Birdshot retinochoroidopathy is a progressive disease with the potential for visual impairment. Patients who present at a later date after the onset of disease were more likely to have vision impairment and CME. Use of long term immunosuppressive therapy may reduce the risk of CME. PMID- 16038651 TI - Electroretinographic monitoring in birdshot chorioretinopathy. AB - PURPOSE: To evaluate electroretinography (ERG) during long-term follow-up in birdshot chorioretinopathy (BCR). DESIGN: Retrospective, comparative interventional case series. METHODS: SETTING: University subspecialty clinic. PATIENT POPULATION: Twenty-three HLA-A29-positive patients with BCR and 40 normal control subjects. INTERVENTION PROCEDURE: Patients were monitored with ERG approximately annually. Treatment was according to best medical judgment. main outcome measures: Baseline ERG values, vision, and ERG values during observed and treated intervals. RESULTS: Median age of patients was 52 years, and 19 patients were untreated at baseline. Eighty-two ERGs were performed. Eighteen patients had more than one ERG; mean follow-up of these patients was 40.2 months +/- 31.2, median 23 months. At baseline, several ERG parameters were statistically reduced compared with control subjects when adjusted for age. The combined rod-cone and cone b/a wave ratios did not differ from control subjects (P = .45 and 0.14). Scotopic rod and combined rod-cone b-wave amplitudes were statistically correlated with baseline vision, as were implicit times for the combined rod-cone a-wave, cone a-wave, and cone flicker b-wave. Median visual acuity was 20/25 and did not change during follow-up. Most ERG parameters showed marked worsening during observed intervals. During treated intervals, the ERG declined at a rate consistent with aging. CONCLUSIONS: Many ERG parameters in patients with BCR greatly differ from control subjects, correlate with vision, and worsen during observation. Selected patients may show improvement in ERG with treatment. The cone b-wave flicker implicit time was most often associated with clinically important measures such as vision, duration of symptoms, and deviation from normalcy. PMID- 16038652 TI - A multi-disciplinary study of the ocular, orthopedic, and neurologic causes of abnormal head postures in children. AB - PURPOSE: To determine the relative frequency that abnormal head postures in children are caused by orthopedic, ophthalmologic, or neurologic disorders, respectively. DESIGN: A prospective, consecutive case series. METHODS: Children found to have an abnormal head posture on routine pediatric examination underwent an evaluation by a pediatric ophthalmologist, pediatric orthopedist, and pediatric neurologist. The study was conducted in northwestern Italy. RESULTS: In the 63 children evaluated, the cause of the abnormal head posture was orthopedic in 35, ocular in 25, and neurologic in 5. In 8 patients, no specific cause could be found. The most common orthopedic cause was congenital muscular torticollis, which accounted for 31 patients. The most common ocular cause was superior oblique muscle palsy, which accounted for 12 patients. In 2 patients neck muscle contracture suggested an orthopedic cause, however, the tight neck muscles were secondary to a head tilt caused by superior oblique muscle palsy. CONCLUSIONS: When the cause of an abnormal head posture is not obvious, a multi-disciplinary approach including ophthalmologic, neurologic, and orthopedic specialists may be helpful. PMID- 16038653 TI - Modern alchemy: fixed combinations of glaucoma drugs. PMID- 16038654 TI - Visual dysfunction after panretinal photocoagulation in patients with severe diabetic retinopathy and good vision. PMID- 16038655 TI - Frailty and age-related macular degeneration: the Beaver Dam Eye Study. AB - PURPOSE: To investigate the associations of measures of frailty to prevalent age related maculopathy (ARM). DESIGN: Cross-sectional population-based study. METHODS: Time to walk a measured course (gait-time), handgrip strength, peak expiratory flow rate, ability to stand from a sitting position without using arms, self-reported co-morbidities, and ARM were assessed at the third examination of the Beaver Dam Eye Study (n = 2,962). ARM was determined by grading stereoscopic color fundus photographs. RESULTS: While controlling for age, smoking, and the number of co-morbid conditions, weaker handgrip strength was associated with early ARM (odds ratio [OR]/10 kg decrease 1.28, confidence interval [CI] 1.08, 1.52, P = .004) and late ARM (OR 1.55, 95% CI 1.02, 2.36) in men but not women. Other measures of frailty were not related to ARM. CONCLUSIONS: A weak cross-sectional association of handgrip strength with ARM (in men) was found after controlling for co-morbid conditions. These data suggest that ARM is due to a specific disease process, albeit age-related, rather than to biologic aging, as reflected by measures of frailty. PMID- 16038656 TI - Intraocular pressure over the clinical range of blood pressure: blue mountains eye study findings. AB - PURPOSE: To quantify the relation of blood pressure (BP) and intraocular pressure (IOP) across the clinical range of BP in an older phakic population not using glaucoma medications. DESIGN: Cross-sectional population-based study METHODS: Seated BP and applanation IOP were measured, and their relation was assessed in regression models. RESULTS: Mean IOP of the two eyes increased linearly from 14.3 mm Hg for systolic BP <110 mm Hg to 17.7 mm Hg for systolic BP > or =200 mm Hg, a 3.4-mm Hg excursion over this range. Mean IOP also increased from 15.2 mm Hg for diastolic BP <70 to 18.4 mm Hg for diastolic BP > or =120, a similar variance. There were no changes after age and multivariate adjustments for IOP or after excluding undiagnosed glaucoma cases. CONCLUSION: A 3-mm linear IOP increase over the clinical spectrum of BP levels was evident; this variance is greater than with most other systemic and ocular parameters. PMID- 16038657 TI - Photodynamic actions of indocyanine green and trypan blue on human lens epithelial cells in vitro. AB - PURPOSE: The purpose of this study was to evaluate the toxicity and photodynamic activity of indocyanine green (ICG) and trypan blue (TryB) on cultured human lensepithelial cells (LECs). DESIGN: Experimental study. METHODS: Lens epithelial cell viability was assessed after treatment with ICG and TryB concentrations ranging from 0.025 to 5.0 mg/ml, and exposure to 806 nm diode laser. RESULTS: At ICG concentrations below 0.5 mg/ml, there was > or =75% cell viability; at higher ICG concentrations there was dose-dependent cytotoxicity in addition to loss of cellular viability due to ICG photosensitization. TryB had little cytotoxicity to the LECs: >80% cells were viable irrespective of the dye concentration or laser treatment. CONCLUSIONS: These data indicate that ICG may have application as a photosensitizer in the selective eradication of residual LECs after cataract surgery to reduce the incidence of posterior capsule opacification. PMID- 16038658 TI - Optical coherence tomography findings in valsalva retinopathy. AB - PURPOSE: To demonstrate the plane of premacular hemorrhage in Valsalva retinopathy with optical coherence tomography DESIGN: Observational case report METHODS: Two young men with Valsalva stress-related premacular hemorrhage were evaluated by OCT scans, taken just above the level of sedimented blood. RESULTS: In both cases, OCT demonstrated two distinct membranes: a highly reflective band immediately above the premacular hemorrhage, corresponding to the internal limiting membrane, and an overlying patchy membrane with low optical reflectivity consistent with posterior hyaloid. CONCLUSION: OCT supported the clinical impression that the plane of premacular hemorrhage in Valsalva retinopathy is probably under ILM. Larger case studies are required to confirm this finding. PMID- 16038659 TI - Autologous cultivated conjunctival transplantation for recurrent viral papillomata. AB - PURPOSE: To describe the use of an autologous serum-free derived cultivated conjunctival epithelial equivalent in the treatment of extensive recurrent viral papillomata. DESIGN: Interventional case report. METHODS: A 10-year-old child with extensive recurrent viral papillomata involving the superior and inferior tarsal, forniceal, and bulbar conjunctiva underwent surgical excision of all diseased areas and double freeze-thaw cryotherapy. Autologous serum-free cultivated conjunctival equivalents were used to reconstruct the ocular surface and conjunctival fornices. RESULTS: Almost complete epithelialization was achieved within 5 days postoperatively. A good cosmetic and functional result was obtained, and no recurrences or cicatricial complications developed during 12 month follow-up. CONCLUSION: Transplantation of autologous cultivated conjunctiva was effectively used in the reconstruction of the ocular surface after extensive excision of recurrent viral papillomata. This modality of treatment may be useful in the treatment of ocular surface disorders in which extensive conjunctival and fornix reconstruction is required. PMID- 16038660 TI - Microkeratome-assisted mushroom keratoplasty with minimal endothelial replacement. AB - PURPOSE: To report the outcome of a new keratoplasty technique aimed at treating full-thickness opacities with minimal removal of recipient endothelium. DESIGN: Interventional case report. METHODS: A deep central scar was removed using microkeratome-assisted mushroom-shaped keratoplasty, consisting of a large anterior stromal lamella (9.0-mm in diameter) and a small posterior button (5.0 mm in diameter) including deep stroma and endothelium. Complete suture removal was performed 3 months after surgery. Visual acuity, refraction, and computerized corneal topography were evaluated pre- and postoperatively. RESULTS: Best spectacle-corrected visual acuity improved from 20/60 to 20/20 at 6 months postoperatively. Postoperative refraction was -2.50-1.00 x 20 degrees. CONCLUSIONS: Microkeratome-assisted mushroom keratoplasty may offer visual and refractive advantages over conventional keratoplasty surgery. Because most of the recipient endothelium is preserved and may spread onto the posterior surface of the small donor button, graft decompensation secondary to immunologic rejection may be avoided with this technique. PMID- 16038661 TI - Ultrasound biomicroscopy in the diagnosis and management of intraocular gnathostomiasis. AB - PURPOSE: To describe the use of ultrasound biomicroscopy in the identification of an intraocular nematode in a case of suspected nematode-induced uveitis DESIGN: Observational case report. METHOD: UBM was performed under topical anesthesia in a patient with acute painful uveitis suspected to result from an intraocular nematode. Clinical examination did not reveal the nematode. RESULTS: Over a 6 minute time span, serial UBM examinations revealed the nematode to move from the iris root into the posterior chamber through the zonules. Subsequently, it was seen adhering to the cornea and could be removed surgically, resulting in symptom relief. CONCLUSION: UBM is a useful tool in diagnosis and management of parasitic uveitis. PMID- 16038662 TI - Downbeating nystagmus and muscle spasms in a patient with glutamic-acid decarboxylase antibodies. AB - PURPOSE: To report the ophthalmic findings and response to treatment in a patient with glutamic-acid decarboxylase antibodies. DESIGN: Case report. METHODS: A 55 year-old woman developed progressive, painful, low back muscle spasms, vertical diplopia, downbeating nystagmus, and asymmetric appendicular ataxia. RESULTS: Downbeating nystagmus was present in primary gaze with an alternating skew deviation in lateral gaze. Serum and cerebrospinal fluid GAD antibodies were detected. Treatment with diazepam led to resolution of spasticity, whereas repeated courses of intravenous immunoglobulin improved cerebellar function, including appendicular ataxia and downbeating nystagmus. CONCLUSIONS: Patients with GAD antibodies may have elements of both Stiff-person syndrome (muscle rigidity and spasms) and prominent cerebellar dysfunction. Treatment with diazepam rapidly improved Stiff-person symptoms, whereas IVIg was partially effective at the early stage of cerebellar dysfunction. PMID- 16038663 TI - Displacement of rectus muscle pulleys by torsional muscle surgery for treatment of full macular translocation-induced incyclotropia. AB - PURPOSE: To elucidate the coronal plane locations of extraocular muscle (EOM) pulleys following torsional muscle surgery. DESIGN: Case report. METHODS: A 76 year-old man underwent advancement of the anterior part of the inferior oblique muscle to treat full macular translocation-induced incyclotropia. Postoperatively, magnetic resonance imaging was used to obtain contiguous, 2-mm thick coronal orbital images. On each MRI image, the cross-sectional area and center of the EOM was computed, and all rectus EOM positions were translated to the coordinate origin at the area centroid of the globe at the level of pulleys. RESULTS: The superior rectus pulley was displaced temporally, and the lateral rectus pulley was displaced inferiorly more than 2 SD from normal subjects. Coronal plane locations of EOM pulleys of the ipsilateral eye showed extorsion compared with that of the contralesional eye. CONCLUSIONS: Torsional muscle surgery causes an extorsional shift of the superior and lateral rectus pulleys. PMID- 16038664 TI - Uveitis in Gaucher disease. AB - PURPOSE: Chronic uveitis has been previously reported in a patient with Gaucher's disease and improved with enzyme replacement therapy. This report describes the course of uveitis in two other patients with type I Gaucher's disease. DESIGN: Observational case reports. METHODS: Review of all patients in a large referral clinic for Gaucher's disease for incidence of uveitis. RESULTS: Two patients with uveitis among 527 patients (0.4%) were found. Each had a different clinical outcome from the other and from the patient cited in the literature. CONCLUSIONS: Both patients have mild Gaucher's disease, but one patient has uveitis well controlled for 10 years solely by local corticosteroids, whereas the other has suffered progression of uveitis despite enzyme replacement therapy. PMID- 16038665 TI - Cystoid macular edema in gyrate atrophy of the choroid and retina: a fluorescein angiography and optical coherence tomography evaluation. AB - PURPOSE: To analyze the importance f optical coherence tomography (OCT) to diagnose the cystoid macular edema in a case of gyrate atrophy. DESIGN: Observational case report. METHODS: A 12-year-old boy presenting with gyrate atrophy of the choroid and retina underwent ophthalmologic, clinical, and laboratory tests. RESULTS: Plasma ornithine level was 735 mumol/l. Fluorescein angiography showed bilateral hyperfluorescence involving the central region of the macula. Optical coherence tomography (OCT) disclosed bilateral intraretinal cysts areas of low reflectivity with occasional high-signal elements bridging the retinal layers and intraretinal thickening. CONCLUSIONS: Both fluorescein angiography and OCT were helpful to confirm the diagnosis of macular involvement as a complication of gyrate atrophy of the choroid and retina in a patient who presented without any clinical evidence of cystoid macular edema, except a decrease in visual acuity. PMID- 16038666 TI - Macular hole in cat scratch disease. AB - PURPOSE: To report the development of a macular hole as a complication of cat scratch disease. DESIGN: Case report. METHODS: A 10-year-old girl was seen with unilateral neuroretinitis from serologically confirmed cat scratch disease. Twelve days later, she developed a macular hole in the involved eye. Fundus photography and optical coherence tomography (OCT) were obtained at presentation and after the development of a macular hole. RESULTS: Fundus photography and OCT on presentation revealed a nasal neurosensory detachment and multiple inflammatory foci throughout the macula, including a subfoveal lesion. Fundus photography and OCT 12 days later revealed the development of a partial posterior vitreous detachment and a full thickness macular hole. CONCLUSIONS: Macular hole should be included among posterior segment complications of cat scratch disease. In this case, the macular hole was associated with partial vitreous detachment and a preexisting subfoveal lesion, likely representing an inflammatory focus. PMID- 16038667 TI - A bent infusion cannula for vitreous surgery. AB - PURPOSE: To modify an infusion cannula for vitreous surgery and decrease cannula related complications. DESIGN: New surgical instrument. METHODS: A conventional 20-gauge infusion cannula was modified by forming a 135-degree angle at the joint of the flange between the cannula adapter and the needle. The sloped plane of the needle tip was opposite the bent adapter. The cannula was routinely fastened with a 7-0 absorbable suture after insertion into the eye. RESULTS: We used the modified infusion cannula in 50 consecutive phakic eyes (48 patients). Intraoperatively, the cannula remained attached to the globe surface; it did not tilt anteriorly and damage the lens when touched inadvertently. No suprachoroidal or subretinal infusion was observed. Neither the lens nor the retina was damaged. There were no cannula-related retinal detachments or breaks during follow-up. CONCLUSIONS: The modified infusion cannula is more effective and safer during pars plana vitrectomy in phakic eyes compared with a conventional cannula. PMID- 16038668 TI - Trabeculotome-guided deep sclerectomy. A pilot Study. AB - PURPOSE: To describe and evaluate a new technique that helps identification and unroofing of Schlemm's canal during deep sclerectomy. DESIGN: A prospective, interventional case series. METHODS: This pilot study was conducted on 15 eyes with various types of glaucoma. After dissecting the superficial scleral flap, the trabeculotome was inserted inside the Schlemm's canal. During deep flap dissection, a direct incision was made over the trabeculotomy to open and unroof Schlemm's canal. Five of the excised deep flaps were submitted for histologic examination. RESULTS: In 13 of the 15 eyes, the Schlemm's canal was properly identified and unroofed. Schlemm's canal endothelium was identified in all the examined specimens. The mean intraocular pressure was reduced from 26.66 +/- 4.54 mm Hg to 12.2 +/- 3.5 mm Hg at the end of a mean follow-up of 9.4 +/- 2.9 months. CONCLUSION: The insertion of the trabeculotome inside Schlemm's canal before dissection of the deep flap helped Schlemm's canal unroofing. PMID- 16038669 TI - Numerical confusion errors in ishihara testing: findings from a population-based study. AB - PURPOSE: To describe the prevalence of numerical confusion errors in Ishihara testing in a representative sample of Australian children. DESIGN: Cross sectional, population-based study. METHODS: The Sydney Myopia Study examined a representative stratified random cluster sample of 1,741 children (aged 6 years) attending 34 schools in Sydney, Australia (response rate, 78.9%), including color vision tests in 1,735 children. Those with any color vision defects (n = 33; 1.9%; 97% boys) were excluded. Responses for each Ishihara plate were recorded. RESULTS: Numerical confusion errors were made by 75.8% of children with normal color vision; there was no gender difference. Plates 3 and 7 were particularly prone to numerical confusion errors; 48.4% and 40.8%, respectively, of children misread these plates. CONCLUSION: Numerical confusion errors in the Ishihara test were relatively common in children with normal color vision. The frequency of such errors could indicate an inherent deficiency in this commonly used test. PMID- 16038670 TI - Migration of intraocular silicone into the cerebral ventricles. AB - PURPOSE: To report a case in which silicone oil in the eye migrated into the cerebral ventricles and the subarachnoid space. DESIGN: Observational case report. METHODS: A 62-year-old woman presented with proliferative diabetic retinopathy and tractive retinal detachment in her left eye. Par plana vitrectomy and injection of 5,000-centistoke-viscosity silicone oil were performed. Intraocular pressure was elevated after the operation and poorly controlled for 4 months. Eight months after the first operation, the patient felt dizzy. Computed tomography (CT), magnetic resonance imaging (MRI), and optical coherence tomography (OCT) scanning were performed. RESULTS: The CT imaging, MRI, and OCT showed silicone oils migrated into the sella cistena superioris and the cerebral ventricles. Follow-up examination was continued. CONCLUSION: Although silicone oil migrating into the ventricles is unusual, we suggest that every patient undergoing silicone oil tamponade with poorly controlled high intraocular pressure and optic disk atrophy should be carefully evaluated. PMID- 16038671 TI - A yellow ring-shaped macular reflection. AB - PURPOSE: To describe a yellow ring-shaped reflection in the macula of healthy subjects observed with a common indirect ophthalmoscope. DESIGN: Observational case series. METHODS: Fundus photographs of the macula were acquired with a slit lamp and 90-diopter lens in 5 healthy subjects (age range, 23-50 years) at a perpendicular angle and at an oblique angle to the retina. RESULTS: The perpendicular fundus photographs showed a yellow ring-shaped reflection with a diameter of approximately 5 degrees. The oblique photographs had a dark red spot in the center. CONCLUSIONS: To our knowledge, this phenomenon has hitherto escaped attention in the literature. We conclude that the shape, size, and location of the yellow reflection are consistent with a cone origin. This observation may be used for diagnostic purposes. PMID- 16038672 TI - Chlamydia conjunctivitis and central retinal vein occlusion. AB - PURPOSE: To report a case of coexistent Chlamydia conjunctivitis and central retinal vein occlusion (CRVO) in which both problems improved significantly after oral doxycyline. DESIGN: Interventional case report. METHODS: A healthy 39-year old man presented with follicular and papillary conjunctivitis, unresponsive to topical antibiotic therapy, and a CRVO. The patient had serum IgG titers of 1:256 to both Chlamydia pneumonia and Chlamydia trachomatous. RESULTS: Treatment with oral doxycyline (100 mg orally twice daily for 2 weeks) resulted in complete resolution of the conjunctivitis and rapid improvement of the CRVO. CONCLUSIONS: Because of its ability to cause chronic vascular endothelial infection and localized inflammation, Chlamydia has been implicated in the development of arterial vascular disease. The positive Chlamydia serology, conjunctivitis, CRVO, and excellent response to doxycycline in this patient suggest that Chlamydia may have contributed to his vascular occlusion. Further studies investigating a possible association between Chlamydia and CRVO may be indicated. PMID- 16038673 TI - Screening for mutations in the IMPDH1 gene in Japanese patients with autosomal dominant retinitis pigmentosa. AB - PURPOSE: To determine the presence and frequency of mutations in the IMPDH1 gene in Japanese patients with autosomal dominant retinitis pigmentosa (ADRP), and to characterize the clinical characteristics of patients with the Lys238Arg mutation in the IMPDH1 gene. DESIGN: Case reports and results of DNA analysis. METHODS: All 14 coding exons of the IMPDH1 gene were directly sequenced in 96 unrelated patients with ADRP. The clinical features were determined by visual acuity, slit lamp biomicroscopy, and kinetic visual field tests. RESULTS: Two novel mutations, a Leu227Pro and Lys238Arg, in the IMPDH1 gene were identified in two unrelated families with ADRP. The clinical features associated with the Lys238Arg mutation were an early-onset and severe retinal degeneration. CONCLUSIONS: The most commonly reported Asp226Asn mutation was not found in the Japanese population, instead two novel mutations were found. These findings suggest that mutations of the IMPDH1 gene cause ADRP in the Japanese population. PMID- 16038674 TI - Retinal nerve fiber layer thickness in the fellow eyes of normal-tension glaucoma patients with unilateral visual field defect. AB - PURPOSE: To quantitatively evaluate retinal nerve fiber layer (RNFL) thickness in the fellow eyes of normal-tension glaucoma (NTG) patients with unilateral visual field defect. DESIGN: Observational case-control study. METHODS: Twenty-nine NTG patients with unilateral visual field defect were enrolled in this study. All 29 fellow eyes showed normal visual field. Thirty-one normal eyes of 31 subjects served as controls. The RNFL thickness around the optic disk was determined using Fast RNFL thickness (3.4) of optical coherence tomography. Average and segmental (4 quadrants and 12 clock- hours) RNFL thickness measurements were compared among the three groups. RESULTS: RNFL thicknesses were significantly different among the three groups in the average, superior quadrant (11 and 12 clock-hour segments), and inferior quadrant (6 clock-hour segment) (P = .00, one-way ANOVA and Tukey's tests). CONCLUSIONS: RNFL thickness reductions are already present in the fellow eyes of NTG patients with unilateral visual field defect. PMID- 16038675 TI - Optical coherence tomography in children. PMID- 16038677 TI - Comparative effect of antiplatelet therapy in retinal vein occlusion evaluated by the particle-counting method using light scattering. PMID- 16038679 TI - Deficient hippocampal neuron expression of proteasome, ubiquitin, and mitochondrial genes in multiple schizophrenia cohorts. AB - BACKGROUND: Hippocampal dentate granule neurons are altered in schizophrenia, but it is unknown if their gene expressions change in schizophrenia or other psychiatric diseases. METHODS: Laser-captured dentate granule neurons from two groups of schizophrenia and control cases and from major depression and bipolar disease cases were examined for alterations in gene expression using complementary DNA (cDNA) microarrays and reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Compared with 24 control cases, the 22 schizophrenia patients in both groups revealed decreases in clusters of genes that encode for protein turnover (proteasome subunits and ubiquitin), mitochondrial oxidative energy metabolism (isocitrate, lactate, malate, nicotinamide adenine dinucleotide [NADH], and succinate dehydrogenases; cytochrome C oxidase; adenosine triphosphate [ATP] synthase), and genes associated with neurite outgrowth, cytoskeletal proteins, and synapse plasticity. These changes were not obtained in 9 bipolar cases or 10 major depression cases and were not associated with age, sex, brain weight, body weight, postmortem interval, or drug history. Brain pH contributed to the variance of some genes but was mostly independent of the disease effect. CONCLUSIONS: Decreases in hippocampal neuron gene expression are consistent with brain imaging and microarray studies of the frontal cortex in schizophrenia. A mitochondrial and ubiquitin-proteasome hypofunctioning of dentate granule neurons may contribute to the deficits of schizophrenia. PMID- 16038680 TI - Subtle fluctuations in psychotic phenomena as functional states of abnormal dopamine reactivity in individuals at risk. AB - BACKGROUND: Subjects at increased risk for psychosis experience continuous variation in the intensity of subtle psychotic experiences in response to minor stressors. It was investigated whether this psychotic reactivity in individuals at risk for psychosis is the exophenotypic expression of an underlying endophenotype characterized by a hyperreactive dopamine (DA) system. METHODS: First-degree relatives (n = 47) and control subjects (n = 49) were studied with the Experience Sampling Method (ESM), a structured diary technique assessing current context and psychopathology in daily life, to assess psychotic experiences in response to stress. A metabolic perturbation paradigm (administration of 2-deoxy-D-glucose inducing a mild state of glucoprivation) causing plasma elevation of homovanillic acid (HVA) was used as a proxy of DA reactivity. RESULTS: Multilevel regression analyses revealed that the interaction between HVA reactivity and daily stress in their effect on psychotic experiences differed according to underlying vulnerability. In the first-degree relatives, underlying HVA reactivity modified the psychotic experiences to daily stress, whereas no such effect was found in control subjects. CONCLUSIONS: These results suggest that psychotic experiences in response to minor stresses in the flow of daily life may be functional states of an underlying abnormal DA reactivity in subjects at risk to develop psychosis. The results add credence to the suggestion that abnormal DA reactivity may be part of the substrate that increases risk for psychotic symptoms in individuals at risk. PMID- 16038681 TI - Neural networks of information processing in posttraumatic stress disorder: a functional magnetic resonance imaging study. AB - BACKGROUND: Neuroimaging studies report reduced medial prefrontal cortical (particularly anterior cingulate) but enhanced amygdala response to fear signals in posttraumatic Stress Disorder (PTSD). We investigated whether anterior cingulate-amygdala dysregulation in PTSD would generalize to salient, but nonthreat related signals. METHODS: Individuals with PTSD (n = 14) and age and sex-matched nontraumatized controls (n = 14) completed an auditory oddball paradigm adapted to functional magnetic resonance imaging at a 1.5-T field strength. RESULTS: Controls displayed bilateral activation in ventral anterior cingulate and amygdala networks, and PTSD subjects showed bilateral dorsal anterior cingulate and amygdala activation to targets relative to nontargets. Compared to controls, PTSD subjects showed enhanced responses to targets in the dorsal and rostral anterior cingulate, and left amygdala. Whole-brain analyses confirmed the expected pattern of distributed prefrontal-parietal responses to targets in the oddball task. Greater activity in posterior parietal somatosensory regions was observed in PTSD. CONCLUSIONS: Our findings of enhanced anterior cingulate responses in PTSD contrast with reports of reduced activity for threat stimuli, suggesting that the latter may be specific to processing of threat related content. Activation in rostral and dorsal anterior cingulate, left amygdala and posterior parietal networks in response to salient, nonthreatening stimuli may reflect generalized hypervigilance. PMID- 16038682 TI - Size versus shape differences: contrasting voxel-based and volumetric analyses of the anterior cingulate cortex in individuals with acute posttraumatic stress disorder. AB - BACKGROUND: Two studies found morphological differences in the anterior cingulate cortex (ACC) of individuals with chronic posttraumatic stress disorder (PTSD). We sought to replicate and extend these findings in a sample of individuals with acute PTSD. METHODS: The ACCs of individuals with acute PTSD (n = 14) and matched healthy control subjects (n = 14) were compared using voxel-based morphometry (VBM), semi-automated volumetric analyses, and probabilistic maps. Posttraumatic stress disorder diagnosis was ascertained by a psychologist using a structured interview. RESULTS: Voxel-based morphometry analyses revealed significantly less gray-matter density in the right pregenual ACC and in the left insula of the PTSD group. However, volumetric analyses of the ACC revealed no significant differences between groups. Probabilistic maps of the labels of the pregenual ACC indicated that the difference between groups in gray matter density was due to shape differences. CONCLUSIONS: Although there are no volumetric differences in the ACC of acute PTSD individuals compared with normal control subjects, significant shape differences exist, which might indicate volumetric differences in the surrounding structures. PMID- 16038683 TI - Emotional dysregulation in adult ADHD and response to atomoxetine. AB - BACKGROUND: Before 1980, attention-deficit/hyperactivity disorder (ADHD) was called minimal brain dysfunction and included emotional symptoms now listed as "associated features" in DSM-IV. Data from two multicenter, placebo-controlled studies with 536 patients were reexamined to assess: 1) the pervasiveness of these symptoms in samples of adults with ADHD; 2) the response of these symptoms to atomoxetine; and 3) their association with depressive/anxiety symptoms. METHODS: The Wender-Reimherr Adult Attention Deficit Disorder Scale (WRAADDS) was used to assess temper, affective lability, and emotional overreactivity, thus identifying patients exhibiting "emotional dysregulation." Other DSM-IV Axis I diagnoses were exclusionary. Outcome measures were the Conners' Adult ADHD Rating Scale (CAARS) and the WRAADDS. RESULTS: Thirty-two percent of the sample met post hoc criteria for emotional dysregulation and had higher baseline scores on ADHD measures, a lower response to placebo, and greater response to atomoxetine (p = .048). Symptoms of emotional dysregulation had a treatment effect (p < .001) at least as large as the CAARS (p = .002) and the total WRAADDS (p = .001). Emotional dysregulation was present in the absence of anxiety or depressive diagnosis. CONCLUSIONS: Symptoms of emotional dysregulation were present in many patients with ADHD and showed a treatment response similar to other ADHD symptoms. PMID- 16038684 TI - Cerebral blood flow changes after treatment of social phobia with the neurokinin 1 antagonist GR205171, citalopram, or placebo. AB - BACKGROUND: Evidence is accumulating that pharmacological blockade of the substance P preferring neurokinin-1 (NK1) receptor reduces anxiety. This study compared the effects of an NK1 receptor antagonist, citalopram, and placebo on brain activity and anxiety symptoms in social phobia. METHODS: Thirty-six patients diagnosed with social phobia were treated for 6 weeks with the NK1 antagonist GR205171 (5 mg), citalopram (40 mg), or matching placebo under randomized double-blind conditions. GR205171 was administered for 4 weeks preceded by 2 weeks of placebo. Before and after treatment, regional cerebral blood flow (rCBF) during a stressful public speaking task was assessed using oxygen-15 positron emission tomography. Response rate was determined by the Clinical Global Impression Improvement Scale. RESULTS: Patients improved to a larger extent with the NK1 antagonist (41.7% responders) and citalopram (50% responders), compared with placebo (8.3% responders). Within- and between-group comparisons showed that symptom improvement was paralleled by a significantly reduced rCBF response to public speaking in the rhinal cortex, amygdala, and parahippocampal-hippocampal regions. The rCBF pattern was corroborated in follow up analyses of responders and subjects showing large state anxiety reduction. CONCLUSIONS: Short-term administration of GR205171 and citalopram alleviated social anxiety. Neurokinin-1 antagonists may act like serotonin reuptake inhibitors by attenuating neural activity in a medial temporal lobe network. PMID- 16038686 TI - Vaccine pharmacotherapy for the treatment of cocaine dependence. AB - BACKGROUND: Cocaine abuse has no established pharmacotherapy, but active immunotherapy with a cocaine vaccine shows promise as a therapeutic intervention. METHODS: An open label, fourteen week, dose-escalation study evaluated the safety, immunogenicity, and clinical efficacy of a novel human cocaine vaccine (TA-CD) in eighteen cocaine dependent subjects. Ten subjects (400 microg total dose group) received four-100 microg injections over the course of eight weeks. Subsequently, eight subjects (2000 microg total dose group) received five-400 microg vaccinations over twelve weeks. Intent to treat analysis of thrice weekly urine toxicologies and cocaine antibody titers were compared. RESULTS: Sixteen of 18 subjects completed the study. There were no serious adverse reactions and the vaccine was well tolerated. The 2000 microg total dose group had a significantly higher mean antibody titer response (2000 units) as compared to the 400 microg total dose group (1000 units) (p = .05). The 2000 microg group was more likely to maintain cocaine free urines than those in the 400 microg group (Z = -3.12, p = .002). Despite relapse in both groups, most reported an attenuation of cocaine's usual euphoric effects at the six month follow-up time points (63% in the 400 microg and 100% in the 2000 microg groups). CONCLUSIONS: The conjugated cocaine vaccine was well tolerated and cocaine specific antibodies persisted at least six months. The likelihood of using cocaine decreased in subjects who received the more intense vaccination schedule. PMID- 16038685 TI - Brain activity in cigarette smokers performing a working memory task: effect of smoking abstinence. AB - BACKGROUND: When nicotine-dependent human subjects abstain from cigarette smoking, they exhibit deficits in working memory. An understanding of the neural substrates of such impairments may help to understand how nicotine affects cognition. Our aim, therefore, was to identify abnormalities in the circuitry that mediates working memory in nicotine-dependent subjects after they initiate abstinence from smoking. METHODS: We used blood oxygen level-dependent (BOLD) functional magnetic resonance imaging (fMRI) to study eight smokers while they performed a letter version of the N-Back working memory task under satiety (< or = 1.5 hours abstinence) and abstinence (> or = 14 hours abstinence) conditions. RESULTS: Task-related activity in the left dorsal lateral prefrontal cortex (DLPFC) showed a significant interaction between test session (satiety, abstinence) and task load (1-back, 2-back, and 3-back). This interaction reflected the fact that task-related activity in the satiety condition was relatively low during performance of the 1-back task but greater at the more difficult task levels, whereas task-related activity in the abstinence condition was relatively high at the 1-back level and did not increase at the more difficult task levels. CONCLUSIONS: We conclude that neural processing related to working memory in the left DLPFC is less efficient during acute abstinence from smoking than at smoking satiety. PMID- 16038687 TI - Leptin treatment in activity-based anorexia. AB - BACKGROUND: Activity-based anorexia (ABA) is considered an animal model of anorexia nervosa (AN). In ABA, scheduled feeding together with voluntary access to a running wheel results in increased running wheel activity (RWA), hypophagia, and body weight loss. Previously it was shown that leptin treatment reduced semi starvation-induced hyperactivity in rats. The present study was performed to confirm and extend this finding, to evaluate leptin's effect on energy balance in ABA. METHODS: The effects of chronic leptin treatment (intracerebroventricular, 4 microg/day) in ABA rats, ad libitum-fed running rats, and sedentary rats exposed to ad libitum feeding or scheduled feeding were investigated. RESULTS: Leptin treatment decreased RWA in ABA rats. Additionally, leptin treatment reduced food intake and increased energy expenditure by thermogenesis in ABA rats. Ad libitum fed running/sedentary rats or food-restricted sedentary rats did not reduce activity after leptin treatment, whereas all leptin-treated rats showed hypophagia. Body temperature was slightly increased in leptin-treated food restricted sedentary rats. CONCLUSIONS: Although leptin treatment reduced RWA in ABA rats, it also prevented hypothermia and decreased food intake. Altogether, this resulted in a stronger negative energy balance and body weight loss in leptin-treated ABA rats. PMID- 16038688 TI - Is radiologists' volume of mammography reading related to accuracy? A critical review of the literature. AB - The current UK quality assurance guidelines for radiologists in the NHS breast screening programme require those reporting screening mammograms to read a minimum of 5000 cases per year. We aimed to review the evidence for this and to assess whether there was justification for lowering the required level. A literature search was conducted to identify relevant studies where accuracy of reporting mammograms was related to reading volume. Three of the five studies reviewed suggested a positive association between reading volume and sensitivity, but there were few data on volumes above 5000 cases per year. The available evidence did not provide any basis for reducing the threshold volume. Further work is needed, in a UK or European setting, to study the relationship between reading volume and accuracy at higher volume levels and also the separate effects of reading volume and reading experience. PMID- 16038689 TI - Bone bruising of the knee. AB - Bone bruising demonstrated by MRI is discussed with histological findings and proposed classifications. The effects of the mechanism of injury on bone bruising at the knee and the natural history of the process are reviewed. The relationship of bone bruising to osteochondral sequelae and to osteoarthritis are considered. PMID- 16038690 TI - Metastatic spinal cord syndromes: imaging appearances and treatment planning. AB - Metastatic spinal cord syndromes usually result from neural compression by adjacent vertebral disease but are occasionally caused by intradural or intramedullary disease. MRI is the most accurate method for evaluation of such syndromes. Knowledge of the relevant imaging appearances and therapeutic options enables the radiologist to make an accurate assessment of the extent of disease and contribute information relevant to treatment planning. PMID- 16038691 TI - MRI of vaginal conditions. AB - Magnetic resonance imaging (MRI) has become an important part of the assessment of suspected vaginal pathology. This pictorial review demonstrates the MRI features and some of the histopathological findings of a variety of vaginal conditions. These may be congenital (total vaginal agenesis, partial vaginal agenesis, longitudinal vaginal septum, transverse vaginal septum), benign (Bartholin's cyst, diffuse vaginal inflammation, invasive endometriosis, ureterovaginal fistula, post-surgical appearances with the formation of a neovagina and adhesions) or malignant, usually due to extension or recurrence from another pelvic malignancy. In this paper, examples of the above are described and illustrated together with examples of the much rarer primary vaginal malignancies. PMID- 16038693 TI - Tuberose sclerosis complex: analysis of growth rates aids differentiation of renal cell carcinoma from atypical or minimal-fat-containing angiomyolipoma. AB - AIM: To study the radiological characteristics of renal masses in individuals with tuberous sclerosis complex (TSC) using serial CT, and to examine how renal cell carcinoma (RCC) may be differentiated from indeterminate cysts or masses. METHODS: This was a retrospective study of 12 cases of TSC in which dedicated renal CT followed after US had demonstrated cystic or sonographically unusual renal masses. The CT density of all masses was measured and the masses categorized as simple cysts, complex cysts, angiomyolipomas or indeterminate solid masses. Subjects were maintained on regular follow-up with repeat CT or MRI and interval renal US. Indeterminate masses that showed rapid growth were considered suspicious for renal cell carcinoma and biopsy or nephrectomy followed. RESULTS: Comparative data were available for a median of 4 years. In each case the renal masses were multiple and bilateral; mean mass diameter was 3.6 cm. Among a total of 206 masses, 18 were simple cysts and 3 were complex cysts. Of the complex cysts, 1 proved to be an angiomyolipoma on histology and the other 2 showed no growth. Of the solid masses, 133 were typical angiomyolipomas (AMLs) and 52 were indeterminate. On follow-up, only 3 indeterminate masses showed rapid growth (>0.5 cm/year), of which only 1 proved to be an RCC on biopsy. The other 2 were minimal-fat AMLs, and the remainder of the masses showed no or slow growth. CONCLUSION: Many renal masses associated with TSC are radiologically indeterminate. A growth threshold of >0.5 cm/year identified the only RCC in this study (0.5% of all masses). Yearly radiological follow-up of indeterminate renal masses is recommended for individuals with TSC. PMID- 16038694 TI - The effect of changing from one to two views at incident (subsequent) screens in the NHS breast screening programme in England: impact on cancer detection and recall rates. AB - AIM: To assess the effect on cancer detection and recall rates of changing from one to two views for incident (subsequent) screens. METHODS: Controlled, comparative, observational study of programmes in NHS breast screening programme in England. SUBJECTS: women aged 50-64 years were screened by the NHSBSP between 1 April 2001 and 31 March 2003. RESULTS: The effect of changing to two-view mammography was a 20% increase in overall incident screen cancer detection rate, with the biggest effect seen for small (<15 mm) invasive cancers. This increased detection rate was achieved with an 11% drop-in recall rate. CONCLUSION: The introduction of two-view mammography for incident screens has resulted in considerable improvements in overall NHS breast screening performance. PMID- 16038695 TI - Radio-guided localization of clinically occult breast lesions (ROLL): a DGH experience. AB - AIM: Wire-guided localization (WGL) of clinically occult breast lesions is a well established technique. The aim of this study was to evaluate radio-guided localization (ROLL) within the breast screening service of a district general hospital. METHOD: The study group comprised 70 women who underwent ROLL under US and stereotaxis. This required an injection of Technetium-labelled colloidal albumen into the impalpable breast lesion. The women then proceeded to theatre, where localization was achieved with the use of a gamma probe. The lesion was identified by the presence of a high signal, caused by the injected isotope. The results of 70 consecutive cases in which a breast lesion was localized using ROLL were compared with the results of the latest 70 WGLs. RESULTS: All 140 lesions were successfully localized. However, the change in technique from WGL to ROLL offered significant benefits to patients. CONCLUSION: Our study demonstrated that ROLL is a practical and reliable localization technique. It can be implemented in hospital units without using valuable gamma camera time. The cost compares well with WGL. There is an improved cosmetic outcome for patients, and the very small quantity of radioactivity used is safe for both patients and staff. PMID- 16038696 TI - Left ventricular aneurysm: comprehensive assessment of morphology, structure and thrombus using cardiovascular magnetic resonance. AB - AIM: To demonstrate that cardiac magnetic resonance (CMR) techniques provide unique and definitive information on the presence, location, size and nature of left ventricular aneurysm and pseudoaneurysm, the presence of thrombus and the viability of myocardium. METHOD: A retrospective study of 27 patients with a final diagnosis of left ventricular aneurysm or pseudoaneurysm, who underwent CMR at a tertiary referral centre in the period between 2000 and 2003. RESULTS: In 7 cases the correct diagnosis of true aneurysm was confirmed; in 7 cases previously unsuspected thrombus was identified; in 7 cases an unsuspected pseudoaneurysm was identified; and in a further 6 cases a previously unsuspected aneurysm was identified. CONCLUSION: CMR refined the diagnosis in the majority of patients with left ventricular aneurysm, and should be considered in all cases of confirmed or suspected left ventricular aneurysm. PMID- 16038697 TI - Postoperative chest radiographic changes after on- and off-pump coronary surgery. AB - AIM: A variety of chest radiograph abnormalities are recognized after coronary artery bypass grafting (CABG). This study analyzes the appearance of preoperative and postoperative chest radiographs in two groups of patients undergoing myocardial revascularization with or without the use of cardiopulmonary bypass (CPB). METHODS: Chest radiographs (preoperative and at 1 day, 6 days and 6 to 8 weeks post operatively) were analyzed according to a detailed protocol in cases of conventional CABG on CPB (n=60) or off-pump coronary artery bypass (OPCAB) (n=60) surgery. On each film 17 different major potential postoperative abnormalities were analyzed. RESULTS: Clinical findings were similar in the two groups. Patients undergoing OPCAB surgery had decreased blood loss, decreased red cell and platelet transfusion, significantly lower intubation time and required less postoperative inotropic support compared with patients undergoing CPB. The CPB group had a significantly higher incidence of left band atelectasis than the OPCAB group (37.6% vs 17.5%, p=0.01) at 6 days postoperatively. There was no other statistically significant difference in any of the 16 remaining major chest radiograph findings between the two groups. CONCLUSION: Although OPCAB surgery is associated with improved clinical outcome compared with conventional CABG surgery, the analysis of postoperative chest radiographs demonstrated only a minor benefit as shown by a reduced degree of left band atelectasis in the OPCAB group. PMID- 16038698 TI - Bile duct complications of hepatic arterial infusion chemotherapy evaluated by helical CT. AB - AIM: To describe the imaging findings of bile duct complications of hepatic arterial infusion chemotherapy (HAIC) using helical CT, to set diagnostic criteria, to develop a CT grading system, and to correlate these with clinical findings and laboratory data. METHODS: Follow-up helical CT of the abdomen was performed every 3 months for 60 patients receiving HAIC. Three radiologists reviewed all CT studies before and after treatment, using either the picture archiving and communication system or hard copies. The findings of bile duct abnormalities were correlated with findings from other imaging techniques, clinical symptoms and laboratory data. RESULTS: Bile duct abnormalities developed in 34 (57%) of cases either during HAIC or 1 to 12 months after treatment. In 14 (41%) of these 34 patients, enhancement of the hepatic parenchyma along the dilated bile duct or in the segmental or lobar distribution was observed. In 43 cases (72%), normal or abnormal alkaline phosphatase levels were consistent with normal or abnormal CT findings, respectively. Increasing alkaline phosphatase and bilirubin levels were related to CT grade. CONCLUSION: Imaging findings of bile duct complications of HAIC are similar to those of primary sclerosing cholangitis, and correlate well with abnormal clinical and laboratory data. In the presence of such clinical abnormalities, thin-section helical CT with careful review of the imaging studies helps to determine the correct diagnosis, monitor the changes and guide appropriate treatment. PMID- 16038699 TI - Radiographers and radiologists reporting plain radiograph requests from accident and emergency and general practice. AB - AIM: To assess selectively trained radiographers and consultant radiologists reporting plain radiographs for the Accident and Emergency Department (A&E) and general practitioners (GPs) within a typical hospital setting. METHODS: Two radiographers, a group of eight consultant radiologists, and a reference standard radiologist independently reported under controlled conditions a retrospectively selected, random, stratified sample of 400 A&E and 400 GP plain radiographs. An independent consultant radiologist judged whether the radiographer and radiologist reports agreed with the reference standard report. Clinicians then assessed whether radiographer and radiologist incorrect reports affected confidence in their diagnosis and treatment plans, and patient outcome. RESULTS: For A&E and GP plain radiographs, respectively, there was a 1% (95% confidence interval (CI) -2 to 5) and 4% (95% CI -1 to 8) difference in reporting accuracy between the two professional groups. For both A&E and GP cases there was an 8% difference in the clinicians' confidence in their diagnosis based on radiographer or radiologist incorrect reports. For A&E and GP cases, respectively, there was a 2% and 8% difference in the clinicians' confidence in their management plans based on radiographer or radiologist incorrect reports. For A&E and GP cases, respectively, there was a 1% and 11% difference in effect on patient outcome of radiographer or radiologist incorrect reports. CONCLUSION: There is the potential to extend the reporting role of selectively trained radiographers to include plain radiographs for all A&E and GP patients. Further research conducted during clinical practice at a number of sites is recommended. PMID- 16038700 TI - CT and angiographic features of hepatic inflammatory myofibroblastic tumour. PMID- 16038701 TI - Hepatocellular carcinoma supplied by the ileocolic branch of the superior mesenteric artery. PMID- 16038702 TI - Repeat survey of current practice regarding corticosteroid prophylaxis for patients at increased risk of adverse reaction to intravascular contrast agents. PMID- 16038703 TI - Repeat survey of current practice regarding corticosteroid prophylaxis for patients at increased risk for adverse reaction to intravascular contrast agents. PMID- 16038704 TI - Imaging patients with "post-cholecystectomy syndrome": an algorithmic approach. PMID- 16038705 TI - Imaging for primary hyperparathyroidism. PMID- 16038706 TI - X-ray dose training: are we exposed to enough? PMID- 16038707 TI - Thrombin increases cardiomyocyte acute cell death after ischemia and reperfusion. AB - Thrombin exerts multiple actions on cardiomyocytes leading to increased intracellular Na+ and Ca2+ concentrations, and to activation of a Ca2+ independent PLA2, and has been proposed to favor the genesis of arrhythmias and ischemic injury in acute coronary syndromes. However, the influence of thrombin on cardiomyocyte cell death during ischemia-reperfusion has not been studied. A beneficial influence of low thrombin concentrations has been described in other cell types. HL-1 cardiomyocytes were subjected to simulated ischemia (SI) and reperfusion (SR) and cell death was assessed by means of LDH release to the incubation media. Thrombin dose-dependently increased cell death in normoxic cells, in cells subjected to SI, and in cells subjected to SR (by 20+/-8%, 95+/ 32% and 35+/-9%, respectively, at 100 U/ml). The effects of thrombin were associated to increased cytosolic Ca2+ overload, mimicked by 100 microM PAR-1 agonist peptide SFLLRNPNDKYEPF, and reversed by the direct thrombin inhibitor lepirudin (IC50=1.3+/-0.2 microg/ml). The presence of thrombin during simulated ischemia-reperfusion increases cardiomyocyte cell death by a mechanism that involves activation of PAR-1 receptors and can be prevented by the direct thrombin inhibitor lepirudin. PMID- 16038708 TI - Myocardial regeneration by endogenous adult progenitor cells. AB - New strategies are urgently needed for the treatment of terminal heart-failure. Recently, new therapeutic strategies were developed to induce cardiac regeneration and thereby enhancing myocardial performance by using the patients own cell pool. In the quest for adequate donor cells skeletal myoblasts, bone marrow (BM) derived cells, endothelial progenitor cells (EPCs) and resident cardiac stem cells but also cytokine application were examined. This review critically discusses the recent findings in the field of endogenous adult progenitor cells and proposes new areas for future research. PMID- 16038709 TI - Probing p53 biological functions through the use of genetically engineered mouse models. AB - The p53 tumor suppressor gene is rendered dysfunctional in the majority of human cancers. To model the effects of p53 dysfunction in an experimentally manipulable organismal context, genetically engineered inbred mice have been the models of choice. Transgenic and knock-out technologies have been utilized to generate an array of different p53 germ line alterations. As expected, many (though not all) of the mutant p53 mouse models are susceptible to enhanced spontaneous and carcinogen-induced tumors of a variety of types. A number of different variables affect the incidence and spectrum of tumors in p53 mutant mice. These include strain background, the nature of the p53 mutation, the presence of wild-type p53 (in addition to mutant p53), exposure to physical and chemical mutagens, or introduction of other cancer-associated genes into the mutant p53 background. In addition to their role in furthering our understanding of the mechanisms of cancer initiation and progression, these models have led to unexpected insights into p53 function in embryogenesis and aging. With the development of ever more sophisticated methods for manipulating the mouse genome, new p53 models are on the horizon, which should deliver advances that will provide not only important mechanistic insights but also discoveries of great clinical relevance. PMID- 16038710 TI - Antiplatelet and anticoagulant therapy in patients with advanced heart failure. PMID- 16038711 TI - Both antiplatelet and anticoagulant therapy may favorably affect outcome in patients with advanced heart failure. A retrospective analysis of the PRIME-II trial. AB - INTRODUCTION: Current guidelines of chronic heart failure (CHF) do not recommend the use of oral anticoagulants (OAC) or antiplatelet therapy (APT). We performed a post-hoc analysis to evaluate the effect of the use of anti-thrombotic therapy with APT and OAC. PATIENTS AND METHODS: We examined 427 patients with advanced CHF, and assessed the effects of the use of APT or OAC at baseline on mortality. We employed a Cox-proportional hazard model to value the effects of APT or OAC use. RESULTS: After a mean follow-up of 3.4 years (range 2.0-5.4), 214 patients died (51%). Forty-one (41) percent (95%CI: 29-53%) of the patients on APT died, and 52% (47-57%) of the patients not on APT (P=0.07). Forty-eight (48) percent (42-54%) of the patients on OAC died, and 55% (46-63%) of the patients not on OAC (P=0.20). This effect of OAC was seen both in patients in sinus rhythm and in atrial fibrillation. After adjusting for important prognostic variables, such as age, LVEF, renal function, and NYHA class, both the use of APT (hazard ratio (HR) 0.62, 95% confidence interval (CI) 0.40-0.97; P=0.04) and the use of OAC (HR 0.60, 95%-CI 0.43-0.83; P<0.01) were related to an improved prognosis. CONCLUSION: This post-hoc analysis suggests that in CHF patients the use of APT or OAC is associated with a higher survival. PMID- 16038712 TI - The G894T polymorphism on endothelial nitric oxide synthase gene is associated with premature coronary artery disease in a Turkish population. AB - INTRODUCTION: The aim of the present study was to investigate the association between premature coronary artery disease and Glu298Asp polymorphism of the endothelial nitric oxide synthase gene. MATERIALS AND METHODS: The eNOS gene polymorphism was analysed in 115 (mean age, 48.1+/-7.9 years) Turkish patients with a diagnosis of premature coronary artery disease and 83 (mean age, 44.6+/ 1.4 years) control subjects. The Glu298Asp polymorphism of the endothelial nitric oxide synthase gene was determined by polymerase chain reaction and restriction fragment length polymorphism. RESULTS: The patients group showed an increase in the frequency of the T allele compared to controls (0.456 versus 0.169, p=0.0001). There was a significant association between the TT genotype and premature coronary artery disease [eNOS TT vs. TG and GG; OR=17.000 (CI 95% 3.952 73.125, p=0.0001)]. The eNOS T/G genotypes were not associated with the number of affected vessels (p>0.05). In addition, the family history of premature coronary artery disease, smoking, diabetes, obesity, dyslipidemia and eNOS TT genotype were independent risk factors of coronary artery disease. The patients with eNOS TT genotype had 15 fold risk of coronary artery disease compared with the control group [OR=15.356(CI 95% 3.262-77.289, p=0.001)]. CONCLUSIONS: These results suggest that premature coronary artery disease is associated with the Glu298Asp polymorphism of the endothelial nitric oxide synthase gene in our population. PMID- 16038713 TI - Superior efficacy of clopidogrel plus acetylsalicylic acid compared with extended release dipyridamole plus acetylsalicylic acid in preventing arterial thrombogenesis in healthy volunteers. AB - INTRODUCTION: Recent ex vivo platelet aggregometry data indicate that clopidogrel 75 mg/day plus acetylsalicylic acid (ASA) 75 mg/day is a more potent antiplatelet regimen than the marketed combination of dipyridamole+ASA. The present study was designed to assess the antithrombotic effect of both dual antiplatelet regimens using a human ex vivo model of arterial thrombosis. MATERIALS AND METHODS: This was a randomized, double-blind, placebo-controlled, crossover study. During two 10-day treatment periods separated by a 14-day washout period, 23 healthy male volunteers received once-daily clopidogrel 75 mg plus acetylsalicylic acid 75 mg, or twice-daily extended-release dipyridamole 200 mg plus acetylsalicylic acid 25 mg. Assessments were made at baseline and on Day 10 of each period. Arterial thrombus formation was induced ex vivo by exposing a collagen-coated surface in a parallel-plate perfusion chamber to native blood for 3 min (arterial wall shear rate 2600 s(-1)). Total platelet and fibrin deposition was determined by immunoenzymatic methods. RESULTS: Compared with baseline values, the mean inhibition of total platelet deposition was 63.9+/-5.9% with clopidogrel plus acetylsalicylic acid, compared with 18.4+/-5.6% for extended-release dipyridamole plus acetylsalicylic acid (67% reduction; 95% CI, 49-79%; p<0.0001). Corresponding figures for fibrin deposition were 64.9+/-4.8% and 18.3+/-9.7%, respectively (58% reduction; 95% CI, 45-67%; p<0.0001). Both treatments were well tolerated. CONCLUSIONS: Clopidogrel plus acetylsalicylic acid showed significantly superior antithrombotic efficacy compared with extended-release dipyridamole plus acetylsalicylic acid in preventing arterial thrombogenesis in humans. PMID- 16038714 TI - Thrombophilic abnormalities and recurrence of venous thromboembolism in patients treated with standardized anticoagulant treatment. AB - INTRODUCTION: Whether patients with hereditary or acquired thrombophilia have an increased risk for recurrence of venous thromboembolism (deep vein thrombosis and/or pulmonary embolism) is still controversial. The aim of this study was to evaluate the incidence of recurrence of venous thromboembolism in patients with and without thrombophilic abnormalities treated with standardized anticoagulant treatment. MATERIAL AND METHODS: Database was from a prospective multicenter randomized study aimed at evaluating the long-term clinical benefit of extending to 1 year the 3-month oral anticoagulant treatment after a first episode of idiopathic proximal deep vein thrombosis. The screening for thrombophilia included antithrombin, protein C, protein S deficiencies, resistance to activated protein C and/or factor V R506Q mutation, the mutation 20210GA of the prothrombin gene, hyperhomocysteinemia and antiphospholipid antibodies. The diagnosis of venous thromboembolism recurrence was done by objective tests and adjudicated by a panel unaware of the results of the thrombophilia screening. RESULTS: A screening for thrombophilic abnormalities was performed in 195 patients. Twenty of 57 (35.1%) thrombophilic patients experienced a recurrence of venous thromboembolism as compared with 29 of 138 (21.0%) patients without thrombophilia (HR=1.78, 95% CI 1.002-3.140, p=0.046). The difference in VTE recurrence between patients with and without thrombophilia was accounted for by those who received 3 months of oral anticoagulation (HR=3.21, 95% CI 1.349-7.616, p=0.008). No difference between thrombophilic and non-thrombophilic patients was observed in the time interval from the index episode to recurrent venous thromboembolism (29.1+/-23.9 and 30.6+/-19.8 months, respectively). CONCLUSIONS: Thrombophilic abnormalities are associated with an increased risk of venous thromboembolism recurrence. The role of thrombophilia in the long-term management of venous thromboembolism should be addressed in prospective management studies. PMID- 16038715 TI - Elevated levels of leukocyte tissue factor mRNA in patients with venous thromboembolism. AB - Tissue factor (TF) mRNA levels in leukocyte and TF antigen in plasma were examined in patients with deep vein thrombosis (DVT). Although TF mRNA levels in leukocytes were higher in patients with DVT than in healthy volunteers, they were lower in patients with DVT than in those with solid cancer and those with disseminated intravascular coagulation (DIC). On the other hand, the plasma levels of TF antigens were markedly high in patients with DVT/pulmonary embolism (PE). Analysis of the role of underlying disease of DVT showed no significant difference in TF mRNA levels and TF antigens among patients with solid cancer, post-surgical, other diseases and those free of underlying diseases. In patients with VTE, plasma levels of D-dimer, soluble fibrin, GE-XDP and plasminogen activator inhibitor-1 did not correlate with TF mRNA or TF antigen. In analysis of 18 patients with PE with and without DVT, TF mRNA levels in leukocytes correlated with the plasma levels of D-dimer. These findings suggest that TF in leukocytes is more likely to be involved in the development of thrombosis in PE than DVT. PMID- 16038716 TI - The influence of the -401G/T and -402G/A polymorphisms of the coagulation FVII promoter on plasma levels of FVII. PMID- 16038717 TI - Correlation of plasma von Willebrand factor levels, an index of endothelial damage/dysfunction, with two point-based stroke risk stratification scores in atrial fibrillation. AB - To test the hypothesis that the new CHADS2 and Framingham point-based risk stratification scores could be related to plasma vWf (a plasma index of endothelial damage/dysfunction) and soluble E-selectin (an index of endothelial activation) levels in a large cohort of AF patients, we studied 200 consecutive patients (101 male; 72+/-9 years) attending our anti-coagulation clinic for the initiation of anticoagulation treatment with acenocoumarol. AF patients had a median CHADS2 score of 2 (1-2) and a median Framingham point-based risk score of 14 (9-21). Results of research indices in our AF patients were as follows: vWf 142.8+/-41.8 IU/dL and sE-sel 44 (31-62) ng/mL. There were significant correlations between plasma vWf levels and both risk CHADS2 and Framingham risk scores (vWf-CHADS2 risk score: Spearman, r=0.249, p<0.001; vWf-Framingham risk score: r=0.294, p<0.001). sE-sel did not show any significant correlation with both risk scores (sE-sel-CHADS2 risk score: r=-0.054, p=0.460; sE-sel-Framingham risk score: r=0.062, p=0.460). There were no statistically significant correlations between vWf and sE-sel (r=-0.127, p=0.081). Both CHADS2 and Framingham risk scores were significantly correlated with each other, r=0.627, p<0.001. In conclusion, in a wide cohort of non-selected and consecutive AF patients, endothelial damage/dysfunction (assessed by plasma vWf levels) but not endothelial activation (sE-sel) correlated with two new risk stratification scores for stroke in AF. Further prospective studies are needed to assess the prognostic role of prothrombotic indices in AF in relation to stroke and thromboembolic events, and their role in complementing clinical risk stratification schemas. PMID- 16038718 TI - Anthocyanins and colonic metabolites of dietary polyphenols inhibit platelet function. AB - Maintenance and achievement of an optimal platelet function by dietary solutions might be considered an interesting target to influence cardiovascular health. Polyphenol-rich foods such as vegetables and fruits have been shown to significantly improve platelet function in ex vivo studies in humans. However, until yet, it still remains unclear if polyphenols itself, their metabolites or a mixture of both are responsible for the beneficial effects observed so far. Our study aims to evaluate the effect of anthocyanins, in vivo metabolites of different polyphenols from colonic origin and a representative mixture of both at physiological concentrations on platelet aggregation and activation function. Some anthocyanins [1 microM], colonic metabolites [10 microM] and a mixture (4 phenolic acid [1 microM], 4 anthocyanin [0.1 microM] showed significant platelet sedating and desensitizing effects. Activation of the platelets (P-selectin expression) was significantly reduced by 10-40% in resting platelets, TRAP activated and hydrogen peroxide-stressed platelets and epinephrine pre-activated platelets relative to controls. The dose-response curve of the weak agonist TRAP was also significantly altered resulting in a >0.8 microM increase of threshold concentration to induce aggregation. The representative mixture was active despite ten times lower concentrations of the individual components, which showed no activity when tested individually at that concentration, indicating synergism of the different components. Platelet reactivity to strong agonists such as collagen and ADP was not influenced. These results show that anthocyanins and in vivo metabolites of polyphenols have anti-thrombotic properties, suggesting themselves as well as their dietary sources and precursors as potential cardiovascular health promoters. PMID- 16038719 TI - Identification of a novel immunodominant cytotoxic T lymphocyte epitope derived from human factor VIII in a murine model of hemophilia A. AB - Gene therapy of hemophilia A could be complicated by the development of immune responses against the vector as well as the Factor VIII (FVIII) transgene. Previous efforts have been focused on identifying FVIII inhibitor antibody epitopes, whereas the cytotoxic T lymphocyte (CTL) epitopes have not been characterized. CTL would kill cells expressing such epitopes and thus limit the efficacy of gene therapy. To investigate CTL responses against human FVIII in a mouse model of hemophilia A, a computer algorithm program (BIMAS) was employed to predict CTL epitopes of human FVIII. The potential binding of these predicted peptides to MHC class I K(b) was evaluated in a TAP-deficient cell line. When recombinant vaccinia virus expressing B domain-deleted human FVIII (vv-FVIII) was used to immunize E16 hemophilia A mice, a specific CTL response against FVIII152 159 was generated. In contrast, a CTL response to four other FVIII peptides was not detected. Therefore, FVIII152-159 represents a dominant CTL epitope. Identification of this epitope raises the possibility that CTL response to FVIII gene-transduced cells can be diminished by deliberatively mutating the dominant CTL epitope while retaining the biologic function of FVIII for hemophilia A gene therapy. PMID- 16038720 TI - A new global assay of coagulation and fibrinolysis. AB - INTRODUCTION: Global clotting assays may reflect an individual's net hemostatic balance and could contribute to prothrombotic and hemorrhagic risk assessment. In this research, a global assay that measures both coagulation and fibrinolytic capacities was developed and investigated. MATERIALS AND METHODS: In the Clot Formation and Lysis (CloFAL) assay, a buffered reactant solution containing trace amounts of calcium, tissue factor, and tissue-type plasminogen activator is added to plasma samples on a 96-well microplate in an automated, thermoregulated (37 degrees C) spectrophotometer. Clot formation and lysis are monitored as continuous changes in absorbance over the course of 3 h. Measurements include maximum amplitude (MA), times to maximum absorbance (T1) and completion of the first phase of decline in absorbance (T2), and area under the curve (AUC), from which a coagulation index (CI) and various fibrinolytic indices (FI) may be calculated. RESULTS AND CONCLUSIONS: MA, T1, and CI were principally influenced by fibrinogen and procoagulant factors. FI was found to be altered by inhibiting activation of plasminogen or thrombin activatable fibrinolytic inhibitor. Median CI was significantly decreased, while FI was markedly increased, in term neonates as compared to healthy adults (CI: 58% vs. 115%, FI: 210% vs. 90%; P<0.001 for each). By contrast, median CI was notably increased, and FI decreased, in healthy pregnant women when compared to adults (CI: 239% vs. 115%, FI: 59% vs. 90%; P<0.001 for each). The CloFAL global assay is analytically sensitive to several key components in the coagulation and fibrinolytic systems, as well as to physiologic alterations in hemostasis. PMID- 16038721 TI - Adenovirus-mediated expression of heparin cofactor II inhibits thrombin-induced cellular responses in fibroblasts and vascular smooth muscle cells. AB - Heparin cofactor II functions as a physiological inhibitor of thrombin activity. The rate of inactivation of thrombin by heparin cofactor II is increased in the presence of dermatan sulfate, which is produced by fibroblasts or smooth muscle cells. To elucidate the role of heparin cofactor II in the extravascular cells, we induced expression of heparin cofactor II in cultured human fibroblasts or vascular smooth muscle cells using adenovirus-mediated gene transfer. After infection of adenovirus vector, these cells secreted heparin cofactor II protein into culture medium. The expressed heparin cofactor II formed the complex with exogenous thrombin and inhibited the proteolytic activity of thrombin. Expression of heparin cofactor II by infection of adenovirus vector inhibited thrombin induced tissue-type plasminogen activator and interleukin-6 releases from fibroblasts and thrombin-induced interleukin-6 release from vascular smooth muscle cells. These findings show that fibroblasts and vascular smooth muscle cells expressing heparin cofactor II are resistant to thrombin-induced cellular responses. PMID- 16038722 TI - The multidrug resistance protein P-glycoprotein and the regulation of chloride channels. PMID- 16038723 TI - CLL immunotoxins. AB - Immunotoxins composed of cell-selective ligands covalently linked to peptide toxins have been developed for the treatment of chemotherapy relapsed or refractory malignancies including chronic lymphocytic leukemia (CLL). A number of CLL immunotoxins have been clinically tested including T101-ricin A, H65-ricin A, DAB(486)IL2, DAB(389)IL2, RFB4 (dsFv)-PE38 and anti-Tac(Fv)-PE38. Remissions have occurred in some patients without significant myelosuppression, but novel agents continue to be needed. Kay and co-workers in this issue of Leukemia Research have targeted interleukin-4 receptors (IL4R) on CLL B cells with a recombinant IL4 pseudomonas exotoxin fusion protein (IL-4(38--37)-PE38KDEL). A fraction of patients (19%) had CLL cells that were extremely sensitive to the immunotoxin. This novel agent may provide an important new therapeutic for use in the treatment of CLL. PMID- 16038724 TI - Adequate cytogenetic examination in myelodysplastic syndromes: analysis of 529 patients. AB - In myelodysplastic syndromes (MDS), the karyotype is one of the most significant prognostic markers with profound impact on differential diagnosis and therapeutic decisions. In a retrospective study, we examined karyotypes of bone marrow specimens of an oligocentric cohort comprising 529 patients with MDS to address the question how many metaphases need to be analyzed to detect even small cell clones with an appropriate expenditure. We found a statistically significant difference of the frequency of normal karyotypes in the patient group with 19 or less analyzed metaphases compared to the group with 20 or more metaphases analyzed (56% versus 47%, p=0.041). Furthermore, we demonstrate that the analysis of 25 or more metaphases can further improve the sensitivity of karyotype analysis and leads to the identification of additional clinically relevant abnormal clones or subclones in a substantial proportion of patients. In summary, our data suggest the examination of at least 20 metaphases in MDS. PMID- 16038725 TI - Increased CD133 expression in bone marrow of myelodysplastic syndromes. AB - Fresh frozen bone marrow biopsies were evaluated immunohistochemically, applying monoclonal antibodies against CD31, CD34, VEGFR-2 and CD133, a novel marker identifying human endothelial progenitor cells (EPCs). Specimens of 51 patients diagnosed with MDS were compared with 16 AML and 18 controls. The percentage of CD34 expressing cells was increased and CD31 expression was decreased in advanced stages of MDS compared with normal BM. VEGFR-2 expression was also raised in MDS. Here we show for the first time that increased numbers of CD133 positive cells are present in the majority of MDS patients. Additionally, those cells occasionally seem to contribute to capillary forming units in bone marrow. PMID- 16038726 TI - Administration of mylotarg 4 days after beginning of a chemotherapy including intermediate-dose aracytin and mitoxantrone (MIDAM regimen) produces a high rate of complete hematologic remission in patients with CD33+ primary resistant or relapsed acute myeloid leukemia. AB - We have used the dose of 9 mg/m(2) of mylotarg 4 days after the beginning of a chemotherapy including intermediate-dose aracytin and mitoxantrone (MIDAM) in 17 patients with refractory (n=4) or relapsed (n=13) AML. Thirteen patients (76%) achieved CR (n=12) or partial CR (n=1). All four refractory patients and all four patients with poor risk cytogenetic achieved CR or CRp. Although the dose of mylotarg given in combination with chemotherapy was not reduced, the toxicity profile was acceptable (1VOD/17 patients). The MIDAM protocol appears to be highly effective especially in patients with poor risk cytogenetic and/or refractory disease. PMID- 16038727 TI - A recombinant IL-4-Pseudomonas exotoxin inhibits protein synthesis and overcomes apoptosis resistance in human CLL B cells. AB - We have determined that CLL B cells consistently express type 3 membrane receptors for the Th2-derived cytokine IL-4 (IL-4R). Furthermore, when added to CLL B cells, IL-4 induces increased apoptosis resistance, increased protein synthesis in CLL B cells and rapid onset activation of STAT1, STAT5 and STAT6. Since the IL-4-IL-4R pathway is intact in CLL B cells and is related to apoptosis resistance, we considered whether we could target this pathway. A recombinant IL 4 Pseudomonas exotoxin fusion protein (IL-4 PE), known to bind to IL-4R, was incubated with CLL B cells. IL-4 PE (10 ng/ml) cultured with CLL B cells resulted in an increase of apoptosis/death from mean levels of 46.6+/-7.0 of non-exposed cells to 69+/-8.6 (n=6). By measuring in vitro protein synthesis, two predominant patterns of sensitivity were observed. In one, CLL B cell clones (n=4) were found to be extremely sensitive to IL-4 PE (IC50's range=6-25 ng/ml). In the second, low concentrations of IL-4 PE induced agonist activity while increasing concentrations induced cytotoxicity in 6 of 21 patient-derived cells. These studies suggest that the IL-4R, on B-CLL cells, can serve as a unique molecular target for directing cytotoxic agents in the therapy of B-CLL. PMID- 16038728 TI - Peripheral T cells of patients with B cell non-Hodgkin's lymphoma show a shift in their memory status. AB - BACKGROUND: Tumor-infiltrating T cells have a positive influence on the clinical course of B cell non-Hodgkin's lymphoma (NHL). T cells in the peripheral blood of patients with B cell NHL, however, have so far rarely been examined. METHODS: Using flow cytometry we examined lymphocyte subpopulations and numbers of naive/memory T cell subtypes among peripheral T cells of patients with B cell NHL (N=22), patients with metastasized solid tumors (N=27), and healthy controls (N=20). In addition, we analyzed the intracellular content of effector molecules granzyme B and perforin and expression of the T cell receptor zeta chain. RESULTS: We observed increased percentages of potentially highly cytotoxic CD8+CD56+ T cells in the peripheral blood of patients with NHL. Both, patients with NHL and patients with solid tumors showed a much higher expression of the chemokine receptors CCR4 and CCR5 on their T cells than healthy controls, suggesting a polarization of their T cells following stimulation with antigen and/or cytokines in vivo. Furthermore, patients with B cell NHL and patients with solid tumors had far lower percentages of naive CD45RA+CCR7+ T cells than healthy controls and, in the case of CD4+ T cells, patients with solid tumors. In contrast, patients with B cell NHL showed markedly increased levels of memory effector CD45RA-CCR7- CD4(+) T cells when compared to healthy controls and patients with metastasized solid tumors. Patients with NHL also showed elevated levels granzyme B within CD8(+) T cells, indicating that the increase in memory effector cells was of functional relevance. CONCLUSIONS: These findings indicate a marked shift in the composition of peripheral T cells of patients with B cell NHL from naive to memory effector-type cells. PMID- 16038729 TI - Major improvement of the reference method of the French drug resistance network for P-glycoprotein detection in human haematological malignancies. AB - The aim of this study was to improve significantly the sensitivity and specificity of the flow cytometric assay of P-glycoprotein (Pgp) implemented and validated by the laboratories of the French Drug Resistance Network [Huet S, Marie JP, Gualde N, Robert J. Reference method for detection of Pgp mediated multidrug resistance in human hematological malignancies: a method validated by the laboratories of the French Drug Resistance Network. Cytometry 1998;34:248-56] in cells displaying low level of resistance. Fluoresceine-conjugated monoclonal antibodies (Mabs) and propidium iodide were respectively replaced by phycoerythrin-conjugated Mabs and Sytox green. The removal of erythrocytes and granulocytes by density gradient was replaced by the lysis of erythrocytes after Mab incubation. Using these conditions, Pgp could be detected in the K-H30 line, which was negative in former studies, with Mab/Control ratios increasing by 3.7- to 5.9-fold, and Mab/Control ratios in the parental sensitive K562 line still ranging between 0.8 and 1.2. When tested on 16 blood samples from patients presenting haematological malignancies, six samples presented low positivity, which was not detected with the former method, while 10 samples remained negative with the two methods. Pgp was specifically detected in pathological blood cells in the six positive samples. PMID- 16038730 TI - Ionic currents in multidrug resistant K562 human leukemic cells. AB - In this study, the expression and functional characterization of currents through the CFTR (cystic fibrosis transmembrane regulator) and ORCC (outwardly rectifying chloride channels) were determined in wild-type K562 chronic human leukemia cells (K562-WT) and in its resistant counterpart, the vincristine resistant cell line (K562-Vinc). Expression of the CFTR and MDR1 (multidrug resistant) gene products was determined by a semi-quantitative RT-PCR protocol. The amplified products in K562-WT and K562-Vinc showed two bands corresponding to CFTR and MDR1. MDR1 mRNA increased by 20-fold in K562-Vinc whereas no change in CFTR mRNA levels was observed. CFTR and ORCC channel activity were measured with a whole cell configuration of the patch clamp technique. Forskolin (40 microM n activator of adenylate cyclase, added to the extracellular side increased the current in both cell lines. A fraction of the activated whole cell currents was inhibited by 500 microM 4,4-diisothiocyanatostilbene-2,2-disulfonic acid (DIDS) and subsequent addition of 500 microM diphenylamine-2-carboxylate (DPC plus DIDS) further inhibited the remaining currents. The levels of forskolin-activated currents and subsequent blockade were similar in both cell lines. The effect of forskolin was prevented in cells previously exposed to 500 microM DPC. The effects of DIDS and DPC on the forskolin-activated whole cell currents support the idea that both CFTR and ORCC are generating a significant fraction of these currents with DIDS inhibiting ORCC currents and DPC inhibiting CFTR currents when the blockers are added one after another to the extracellular side. Finally, we show that exposure of K562 cells to vincristine which results in the over expression of MDR1 is not accompanied by a significant down regulation of CFTR as in other cells. PMID- 16038731 TI - Expression of heat-shock proteins is associated with major adverse prognostic factors in acute myeloid leukemia. AB - To identify prognostic factors alternative or additional to drug-resistance and apoptosis proteins, we studied the impact of the expression of heat-shock proteins (HSPs) in 98 newly diagnosed acute myeloid leukemia (AML). HSP27 was expressed by 39%, HSP60 by 26%, HSP70 by 58%, HSP90 by 41%, and HSP110 by 30% of cases. HSP expressions were correlated with that of differentiation antigens (CD34, CD14, CD15, CD33) and that of drug-resistance (MRP, MRK) and apoptosis (Bcl-2) proteins. HSP90 and HSP110 were correlated with FAB subtype and karyotypic grouping. Complete remission (CR) was obtained in 68 cases (69%). Median disease-free survival (DFS) of the 68 remitters was 18.1 months with a 3 year DFS rate of 41%. CR rates were higher in patients with lower expression of HSPs. Overall survival (OS) was significantly longer in patients with lower expression of HSPs. Cytogenetics, CD34 positive expression, MRK positive expression, and HSP110 positive expression remained as pejorative prognostic factors for OS in the multivariate analysis. When considering patients with intermediate risk cytogenetics, HSP110 and MRP positive expressions and CD33 negative expression were of poor outcome, while HSP27 and HSP60 positive expressions appeared of pejorative prognostic value in patients with unfavorable karyotypes. PMID- 16038732 TI - 2-Methoxyestradiol at low dose induces differentiation of myeloma cells. AB - Previous studies showed that 2-methoxyestradiol (2ME2) could suppress the proliferation of myeloma cells and induce their apoptosis. In the present study, we found that treatments with low-concentration of 2ME2 resulted in some maturing morphological changes of myeloma cells. Flow cytometric analysis showed that the expression of CD49e on the myeloma cells surface was significantly increased by 2ME2. Moreover, 2ME2 increased the secretion of light chain protein remarkably. Furthermore, the expressions of transcription factor XBP-1 mRNA and protein were also up-regulated. These results demonstrated that 2ME2 at low-concentration could induce differentiation of the myeloma cells, which would provide a new, safe strategy for myeloma therapy. PMID- 16038733 TI - Development of two cytogenetically abnormal clones from multipotential hematopoietic stem cells in a patient with juvenile myelomonocytic leukemia. AB - We report a patient with juvenile myelomonocytic leukemia who had two cytogenetically independent clones at the time of diagnosis. Fluorescence in situ hybridization analyses showed that 42.5% of myeloperoxidase(+) cells and 27.3% of CD20(+) cells had three signals for chromosome 8, while 13.1% of myeloperoxidase(+) cells and 6.0% of CD20(+) cells had del (Y). However, a great majority of CD3(+) cells had no numerical aberration of the two chromosomes. The two karyotypically abnormal clones might have developed from multipotential hematopoietic stem cells with the potential to differentiate into myeloid and B lymphoid lineages, but not T-lymphoid lineage. PMID- 16038734 TI - Assessment and follow-up of the proportion of T315I mutant BCR-ABL transcripts can guide appropriate therapeutic decision making in CML patients. AB - Quantitative monitoring of imatinib mesylate (IM)-resistant, mutated BCR-ABL(+) cells during the follow-up of CML could be useful for optimizing therapeutic management. We retrospectively analyzed T315I mutated BCR-ABL clones throughout the CML history of two patients by nested-PCR-RFLP. At the time of progression, the T315I mutation represented 100% of the BCR-ABL transcripts. During follow-up, we showed that (i) despite a molecular response to IM, a high proportion of T315I transcripts were present (>85%) and predictive of relapse, (ii) interruption of IM and switching to other therapies resulted in a significant reduction in mutant transcript level while total BCR-ABL(+) transcripts remained stable. PMID- 16038735 TI - In vitro activity of the flt3-inhibitor su5614 and standard cytotoxic agents in tumour cells from patients with wild type and mutated flt3 acute myeloid leukaemia. AB - The correlation between drug sensitivity in vitro and the mutation status of the FLT3 receptor gene was evaluated in tumour cells from 17 previously untreated AML patients. Tumour cells with internal tandem duplication (ITD) in the FLT3 receptor gene were significantly more sensitive to the FLT3 inhibitor SU5614 than tumour cells with wild type FLT3. Combinations of SU5614 with etoposide and amsacrine showed better effect (p<0.05) compared with the respective single drugs. Our results suggest that the FLT3 inhibitor SU5614 may have a therapeutic potential, especially in combination with other cytotoxic agents, in patients with FLT3-ITD positive AML. PMID- 16038736 TI - Persistence of various chromosomal aberrations in recipient cells during complete remission after bone marrow transplantation followed by graft rejection. AB - A 16-year-old boy in a second remission of acute lymphoblastic leukemia (ALL) had undergone transplantation of bone marrow from an unrelated donor. The conditioning regimen consisted of high-dose cytarabine, etoposide and 12 Gy of total-body irradiation. Although the donor marrow was rejected, hematopoiesis by the recipient himself recovered and he has remained in complete remission for more than 8 years after stem cell transplantation (SCT). Bone marrow karyotype analysis 1 month after SCT showed random chromosomal aberrations. Although complete remission was maintained, various chromosomal aberrations were detected in marrow cells, and in peripheral blood cells under phytohemagglutinin stimulation over 8 years. Moreover, a clone including del(20)(q11) appeared in marrow cells 7 months after SCT and thereafter was also detected 5 years later in the peripheral blood. This persistence of various chromosomal aberrations and a stable clone without evolution to myelodysplastic syndrome or leukemia support the multi step theory of leukemogenesis. PMID- 16038737 TI - Late ovarian relapse of TEL/AML1 positive ALL confirming that TEL deletion is a secondary event in leukemogenesis. AB - We describe here a late extramedullary ovarian relapse in an 18-year-old female who was diagnosed with hypotetraploid cell acute lymphoblastic leukaemia (cALL) at the age of 6. At both occurrences of the disease cells were analyzed by morphology, immunophenotyping, cytogenetics and molecular methods. TEL/AML1 was detected by RT-PCR and FISH analysis in both events. We demonstrated, using detection of IGH/TCR rearrangements and TEL/AML1 breakpoints sequencing that the cells were clonally related. Moreover, interphasic FISH using TEL and AML1 probes showed the loss of a second TEL at the time of relapse. This observation confirms that TEL/AML1 alone is not sufficient to trigger ALL and that TEL deletion is a secondary event in leukemogenesis. To our knowledge, it is the first complete description of extramedullary ALL relapse combining all methodologies. PMID- 16038738 TI - Bone marrow cells of myelodysplastic syndromes exhibit significant expression of apollon, livin and ILP-2 with reduction after transformation to overt leukemia. PMID- 16038739 TI - Imatinib therapy in Hypereosinophilic Syndrome: a case of molecular remission. PMID- 16038740 TI - Group A streptococcal meningitis: case report and review of the literature. AB - Group A streptococcus is an uncommon cause of meningitis in children. We report a single case of Group A streptococcus meningitis, in an apparently healthy 6-week old infant. Twenty-five cases in the English-language literature in the last 25 years and our case are reviewed. The commonest associated illness was otitis media. Of the 25 patients, 24 survived. Antibiotic therapy, which consisted of penicillin in the majority of patients, was effective. Ceftriaxone was an alternative agent. Neurological sequelae were not uncommon. This report emphasizes the fact that Group A streptococcus can cause meningitis in healthy children without apparent recognizable foci of infection. PMID- 16038741 TI - Fatal Kytococcus schroeteri bacteremic pneumonia. PMID- 16038742 TI - Idiopathic Cd4+T-lymphocytopenia--two unusual patients with cryptococcal meningitis. AB - Idiopathic CD4+T-lymphocytopenia (ICL) is a syndrome characterised by the depletion in the CD4+T-cells but without evidence of HIV infection. Aside from low CD4+lymphocyte counts, the immunologic findings in these patients are distinct from the abnormalities found in HIV infection. There are numerous reports of ICL associated with different diseases and clinical conditions. Opportunistic infections, mostly seen in HIV patients are the most common among them. We describe two patients without risk factors for human immunodeficiency virus (HIV) infection, each of whom presented with cryptococcal meningitis and was found to have idiopathic CD4+T-lymphocytopenia. One of them also acquired EBV and CMV coinfection of the central nervous system. PMID- 16038743 TI - A case of Haemophilus aphrophilus and Mobiluncus mulieris hepatic abscess. PMID- 16038744 TI - A case report of pityriasis lichenoides in a patient with chronic hepatitis C. AB - The authors describe a case of chronic hepatitis C associated with pityriasis lichenoides. The association and evolution during antiviral treatment of these two diseases in this patient point to a possible pathogenetic link between chronic C virus infection and pityriasis lichenoides. PMID- 16038745 TI - Multiple abscesses caused by Gardnerella vaginalis in an immunocompetent man. AB - Gardnerella vaginalis causing significant infection in men is rare. We report a case of sepsis in a previously well man, who developed a perinephric abscess and empyema. G. vaginalis was isolated after prolonged culture of samples from both sites. The microbiological features of the case are discussed. PMID- 16038746 TI - Regression of subcorneal pustular dermatosis type of IgA pemphigus lesions with azithromycin. AB - We present our experience with a 56-year-old man who complained for generalized dermatopathy, manifested by skin lesions with diameter from 0.5 to 5 cm. The lesions did not respond to a 20-day systemic steroid regimen. The results of biopsies of three excised lesions, in combination with the clinical manifestations, led to the diagnosis of subcorneal pustular dermatosis type of IgA pemphigus. An unexpected improvement was noted after treatment with azithromycin (which was provided for management of concurrent non-specific urethritis) and local non-potent steroid plus keratolytic agent ointment. PMID- 16038747 TI - Whipple's disease: multiple hospital admissions of a man with diarrhoea, fever and arthralgia. AB - Whipple's disease is a rare chronic multi-systemic infectious disorder caused by the Gram-positive bacillus, Tropheryma whippelii. Infection may involve any organ in the body, and most commonly affects white men in the fourth to sixth decades of life. The most common presenting symptoms are gastrointestinal and include abdominal pain, diarrhoea, anorexia and associated weight loss. However, the variability in presentation is considerable and some patients may present with intermittent low-grade fever, neurological abnormalities (nystagmus, ophthalmoplegia, cranial nerve defects), migratory arthralgia, lymphadenopathy, or involvement of the cardiovascular system. In typical Whipple's disease, the most severe changes are seen in the proximal small intestine and biopsy reveals mucosal and lymph node infiltration with large, foamy histocytes, containing granules that stain positive with periodic acid-Schiff (PAS) reagent and represent intact or partially degraded bacteria. Extended antibiotic treatment (up to 1-year) is indicated. Life-long surveillance for recurrence is essential, once primary treatment has been completed. We report the case of a 58-year-old man who developed a rare infection with the actinobacterium, T. whippelii. The patient had suffered intermittent episodes of varying clinical symptoms associated with multiple hospital admissions and clinical diagnoses, spanning a period of 22 years. Historically, arthralgia was the primary manifestation in this patient and also was the chief complaint for which he was first hospitalized 22 years ago. At his most recent admission to our hospital department, his presenting symptoms were persistent fatigue, weight loss, arthralgia and diarrhoea. Thus, it is essential that clinicians retain a high index of suspicion for T. whippelii infection in patients who have a long-term history of arthritis, fever and diarrhoea. PMID- 16038748 TI - Actinomycotic infection of the oesophagus. AB - Actinomycotic infections involving the oesophagus are uncommon but have been reported in both immunocompromised and immunocompetent individuals. We report a case of actinomycosis oesophagitis in a patient with lung cancer who received chemo- and radiotherapy. This patient was admitted with severe dysphagia and odynophagia and biopsy from an oesophageal ulcer found on oesophagogastroduodenoscopy (EGD) revealed actinomycosis. The patient was treated with intravenous penicillin G followed by ceftriaxone with clinical improvement and repeat EGD showed reduction in the size of the oesophageal ulcer, but he relapsed due to non-compliance. We review the English literature regarding the clinical features, diagnosis, and management of actinomycotic infections of the oesophagus. PMID- 16038749 TI - Serratia marcescens prosthesis infection successfully treated with meropenem after imipenem failure. AB - A 78-year-old woman developed an early knee-prosthesis infection due to multiresistant Serratia marcescens that was successfully treated with high-dose meropenem, after failure of a long-term therapy combining imipenem and multiple surgical interventions. Because of its lower neurotoxicity, meropenem might be preferred to imipenem/cilastatin for the treatment of osteo-articular infections due to multiresistant Gram-negative bacilli in the elderly. PMID- 16038750 TI - Pneumococcal meningitis and etanercept--chance or association? AB - Rheumatoid arthritis is a severe deforming chronic disease which has major implications for mortality and quality of life. Agents with anti-tumour necrosis factor alpha (TNFalpha) activity are a new modality of therapy, which can significantly reduce the acute inflammation in this condition. However, TNFalpha is a cytokine involved in initiating the protective immune response; consequently, patients receiving this therapy are at increased risk of infection. Etanercept is a recombinant form of the p75 TNF receptor (TNF-RII) dimerised by fusion with a portion of the human IgG1 Fc tail with anti-TNFalpha activity. We report the first case of a patient with rheumatoid arthritis who developed pneumococcal meningitis whilst on etanercept, suggesting a possible association between etanercept and this severe life threatening infection. PMID- 16038751 TI - Vertebral osteomyelitis due to Fusobacterium species: report of three cases and review of the literature. AB - We describe three cases of Fusobacterium spp. diskitis and review with attention to risk factors, clinical features, diagnosis, treatment and outcome. In most of the reported cases, a ear-nose-throat infection was found. Clinical manifestations were similar to those of classic bacterial vertebral osteomyelitis. Clindamycin is the most appropriate antibiotic. The outcome seems to be very good without relapse with appropriate treatment compared to pyogenic vertebral osteomyelitis. PMID- 16038752 TI - Efficacy of plasma exchange and methylprednisolone pulse therapy on influenza associated encephalopathy. AB - Patients with a new type of influenza-associated encephalopathy with high mortality are increasing in Japan and the United States. We present three patients treated with methyprednisolone pulse treatment and plasma exchange to remove cytokines, and all three patients recovered without severe sequela. IL-6 decreased dramatically after the start of the plasma exchange and methyprednisolone. Therefore when influenza-associated encephalopathy is actually diagnosed, steroid pulse therapy should be started at an early stage, and when signs of DIC and/or MOF appear, plasma exchange is recommended to remove the cytokines and NOx. PMID- 16038753 TI - Typhoid fever complicated by multiple organ involvement: report of two cases. AB - Typhoid fever complicated by multiple organ involvement has been rarely mentioned in the literature. We reported two cases of typhoid fever with several unusual manifestations, including acute renal failure, acute hepatitis, acute pancreatitis, disseminated intravascular coagulation, and lower gastrointestinal bleeding. A renal biopsy in the first case showed no pathological change. Bone marrow biopsy showed focal necrosis of matrix, which might have been due to severe illness. A liver biopsy in the second case showed a predominantly histiocytic proliferation with occasional neutrophilic infiltration in the portal areas and hepatic sinusoids. Focal necrosis, bile duct injury, and multiple eosinophilic bodies were also noted. After appropriate antimicrobial therapy, both patients recovered without any sequelae. The potential of multiple organ involvement is highlighted in typhoid fever, which, on rare occasions, may occur simultaneously in the same patient. PMID- 16038754 TI - Acute renal failure and Fanconi syndrome in an AIDS patient on tenofovir treatment--case report and review of literature. AB - Tenofovir disoproxil fumarate, a prodrug of tenofovir, is a potent nucleotide analogue reverse-transcriptase inhibitor with activity against human immunodeficiency virus (HIV). Although initially thought to be relatively safe with regards to nephrotoxic effects compared to its class drugs-adefovir and cidofovir, several cases of acute renal failure and proximal tubule dysfunction have been described in the last few months. We report another patient who developed Fanconi syndrome while on tenofovir. Her condition improved on discontinuation of the drug. We also review the literature of all patients who have developed Fanconi syndrome on tenofovir. PMID- 16038755 TI - Mild leptospirosis with three-year persistence of IgG- and IgM-antibodies, initially manifesting as carpal tunnel syndrome. AB - Long-term persistence of IgG- and IgM-antibodies against leptospira after mild leptospirosis has not been reported. In a 45-year-old female pet-shop worker with carpal tunnel syndrome, accompanied by arthralgias, coughing, repeatedly elevated temperature, followed by easy fatigability, personality change, memory and speech disturbance, blurred vision, myalgia and swollen lymph nodes, leptospirosis was diagnosed, based upon history, clinical findings, and serological investigations. After the described symptoms had disappeared following doxycyclin for 2 weeks, IgG- and IgM-antibodies against leptospira remained positive during the next three years. This case illustrates that leptospirosis may start as carpal tunnel syndrome and that the severity of leptospirosis does not seem to be related to the intensity of the humoral immune response against the causative agent. PMID- 16038756 TI - Cutaneous abscess due to Eubacterium lentum in injection drug user: a case report and review of the literature. AB - We described the first case, to the best of our knowledge, of cutaneous abscess due to Eubacterium lentum in a parenteral drug user, after complete fracture of the right femor. The case underlines the importance of carefully performed microbiological tests, due to the peculiar cultural needs of the micro-organism. PMID- 16038757 TI - Alkhumra virus infection, a new viral hemorrhagic fever in Saudi Arabia. AB - OBJECTIVES: Four patients with typical acute viral hemorrhagic fever were identified in the holy city of Makkah, Saudi Arabia, between 8 and 23 February 2001, the Hajj (pilgrimage) period of that year. Tests for Rift Valley fever (RVF), Crimean-Congo hemorrhagic fever (CCHF), and dengue were negative. Blood specimens were sent to the Centres for Disease Control and Prevention (CDC), Atlanta for viral culture and testing for other hemorrhagic fever viruses. A new flavivirus closely related to the tick-borne Kyasanur forest disease virus was isolated. This new flavivirus was originally isolated in 1995 from 6 patients with dengue-like hemorrhagic fever from Alkhumra district, south of Jeddah, Saudi Arabia. METHODS: A case definition was formulated for surveillance of this new disease in Saudi Arabia. Blood specimens were collected from all patients with suspect 'Alkhumra' virus (ALKV) infection and tested for ALKV, RVF, CCHF, dengue, and West Nile encephalitis. Patients data were prospectively collected on standardized data collection forms. RESULTS: From 8 February 2001 through 9 February 2003, a total of 37 cases were identified in Makkah, 20 of them were laboratory confirmed. Acute febrile flu-like illness with hepatitis (100%), hemorrhagic manifestations (55%), and encephalitis (20%) were the main clinical features. The case fatality was 25%. The disease seemed to be transmitted from sheep or goat to humans by the mosquito bites or direct contact with these animals. CONCLUSIONS: ALKV infection is a novel serious zoonotic hemorrhagic fever virus discovered in Saudi Arabia. The role of arthropods such as ticks and mosquitoes, and animals such as sheep, goat, and rodents in the transmission and maintenance of the virus remains to be elucidated. PMID- 16038758 TI - The use of corticosteroid as treatment in SARS was associated with adverse outcomes: a retrospective cohort study. AB - OBJECTIVE: To study the effect of corticosteroids in the treatment of severe acute respiratory syndrome (SARS). METHODS: A retrospective cohort of 78 consecutive adult SARS patients admitted to a regional hospital in Hong Kong between March and May 2003 was analysed to study the effectiveness of corticosteroid. They were categorized according to whether or not corticosteroid therapy was given, and compared in terms of demographic characteristics, comorbidities, peak lactate dehydrogenase (LDH) levels and clinical outcomes. Established adverse prognostic factors including old age, comorbidities and high LDH levels were used as covariates in multiple logistic regressions to adjust for their confounding effect on adverse outcomes. RESULTS: Among 78 patients, 66 patients (84.6%) received corticosteroid. The LDH level was similar in both groups. The corticosteroid group had more adverse outcomes (37.9% vs. 16.7%) despite younger age and less comorbidity. In multivariate analysis, corticosteroid treatment was associated with a 20.7-fold increase in risk of either ICU admission or mortality, independent of age and disease severity. CONCLUSION: Despite more favourable baseline characteristics and similar peak LDH levels, SARS patients given corticosteroid had more adverse outcomes. PMID- 16038759 TI - The occurrence of influenza A virus on household and day care center fomites. AB - OBJECTIVE: The goal of this study was to evaluate the prevalence of influenza A virus on surfaces in day care and home settings to better assess the potential role of fomites in the transmission of influenza. METHODS: During two and a half years, 218 fomites were tested from 14 different day care centers. Ten different fomites from bathrooms, kitchens and play areas were sampled. In addition, 92 fomites from eight different homes with children were tested over 6 months. Fourteen different household fomites from bathrooms, kitchens and living areas were sampled. Influenza A viral RNA was detected using reverse transcriptase polymerase chain reaction. RESULTS: Influenza was detected on 23% of day care fomites sampled during the fall and 53% of fomites sampled during the spring. Spring and fall sample data was determined to be statistically different at the 0.05 alpha-level by Chi-square analysis P<0 and Fisher's Exact test P=0.00002. There was no statistical difference found between moist and dry fomites (Chi square P=0.13998). No influenza was detected on home fomites sampled during the summer. In contrast, influenza was detected on 59% of home fomites sampled during March. CONCLUSIONS: Influenza A virus was detected on over 50% of the fomites tested in homes and day care centers during influenza season. PMID- 16038761 TI - Pulmonary involvement in brucellosis. AB - OBJECTIVES: Pulmonary involvement is a rare manifestation of brucellosis. The aim of this study was to determine the incidence and forms of pulmonary involvement in the course of brucellosis. METHODS: A prospective study was carried out in 110 patients with brucellosis. All the patients were evaluated with their pulmonary symptoms, physical examination and chest radiography. If pulmonary pathologic findings were present, patients underwent additional diagnostic evaluations including computerized tomography of the thorax and pulmonary function tests. RESULTS: From 110 patients, 11 (six females and five males) were diagnosed as pulmonary brucellosis. Eight of 11 patients had pulmonary symptoms including cough, sputum and dyspnoea. Radiologic findings were parenchymal nodules, lobar pneumonia, paratracheal lymphadenopathy and pleural effusion. At the end of the treatment of brucellosis, clinical findings of pulmonary involvement were recovered in all patients except four dyspnoeic patients who had coexisting COPD. Radiological findings were normal in three and improved in four patients after 6 months of the treatment. CONCLUSIONS: Pulmonary involvement is a rare event in the course of brucellosis. But especially in endemic regions, brucellosis should never be forgotten as a causative agent in patients with pulmonary symptoms. PMID- 16038760 TI - SEN virus infection among patients on maintenance hemodialysis in southern Taiwan. AB - OBJECTIVES: The prevalence and clinical significance of two SEN virus (SENV) strains, SENV-D and SENV-H, was investigated among patients on maintenance hemodialysis in southern Taiwan. METHODS: Sera from 99 uremic patients were tested for alanine aminotransferase (ALT), hepatitis B surface antigen (HBsAg), hepatitis C virus antibody (anti-HCV), and SENV-D and -H DNA. Sera from 120 healthy blood donors were tested as a control group. RESULTS: Of the 99 patients, 24.2 and 7.1% were positive for anti-HCV and HBsAg, respectively, and the prevalence of SENV-D and/or -H (SENV-D/H), SENV-D and SENV-H DNA was 61.6, 46.5 and 27.3%, which was significantly higher than for donors (23.3, 18.3, and 5.8%, respectively; all P<0.0001). Patients with SENV-D DNA had significantly higher mean age among both patients (P=0.019) and donors (P=0.015). Mean serum ALT levels were significantly higher among patients with anti-HCV (P=0.003) and patients with SENV-D and -H concurrent infection (P=0.034). By multivariate analyses, the presence of serum anti-HCV and concurrent SENV-D and -H infection were independent factors associated with increased ALT level. CONCLUSIONS: The prevalence of SENV-D/H infection is high among Taiwanese patients on maintenance hemodialysis with SENV-D more prevalent than SENV-H. The ALT level was associated with not only positive anti-HCV but also concomitant viremia of both SENV-D and SENV-H. PMID- 16038762 TI - Acute flaccid paralysis: the spectrum of a newly recognized complication of West Nile virus infection. AB - OBJECTIVES: Acute flaccid paralysis (AFP) has recently emerged as a major central nervous system complication associated with West Nile virus (WNV) infection. The spectrum of clinical presentations of AFP in WNV infection and its sequelae have not been well-studied. METHODS: We describe three patients with AFP due to WNV infection and review the clinical presentations of 56 patients with this complication derived from published studies. RESULTS: Patients with AFP and WNV presented with a spectrum of illness ranging from single extremity paralysis to quadriparalysis with cranial nerve involvement. Patients commonly developed respiratory failure (54%) and bladder dysfunction (22%). While fever was nearly universal (92%), signs of meningismus were less common (17%). Cerebrospinal fluid (CSF) analysis generally revealed a modest pleocytosis, and imaging studies were not diagnositic. Persistent neurologic impairment occurred in all survivors; overall mortality rate was high (22%) and was associated with both the extent of paralysis and advanced age. CONCLUSION: AFP in the setting of WNV is associated with significant mortality and long-term morbidity. PMID- 16038763 TI - Antibiotic use in acute cholecystitis: practice patterns in the absence of evidence-based guidelines. AB - OBJECTIVES: Antibiotics are frequently administered in acute cholecystitis for preoperative prophylaxis or postoperative treatment. The optimal timing, choice, and duration of antibiotics are unclear. METHODS: We conducted a retrospective review of all cases of acute cholecystitis between 1996 and 2001 at the American University of Beirut Medical Centre. A survey among general surgeons was also performed to describe the pattern of antibiotic prescribing in uncomplicated acute cholecystitis. A MEDLINE search for guidelines for antibiotic use in acute cholecystitis was conducted. RESULTS: The number of cases of acute cholecystitis was 79. The mean duration of postoperative antibiotic therapy was 5 days. There was no correlation between the severity of symptoms, gallbladder description, or positive gallbladder culture and the use of antibiotics postoperatively. Sixty five percent of interviewed surgeons would continue antibiotic therapy postoperatively for 3 or more days. Search of the medical literature failed to provide clear guidelines for antibiotic use in acute cholecystitis. CONCLUSIONS: The use of antibiotics in patients with acute cholecystitis is erratic and costly. Prospective studies are needed to better study the effectiveness of a short course of antibiotics in uncomplicated cases. The role of gallbladder culture in guiding antibiotic therapy should be defined as routine cultures add to the cost without evident benefit. PMID- 16038764 TI - Irrational use of antibiotics among university students. AB - STUDY OBJECTIVE: The aim of the presented study was to evaluate the knowledge, attitude and behaviour of antibiotic usage in the student groups of a University in a country where the antibiotics are taken without prescriptions. DESIGN: Out of 5345 students (excluding those from the Faculty of Medicine) of Ege University, 678 were chosen by systematic sampling methods. The sampling group was divided into two groups. Group A included students from the Faculties of Pharmacy and Dentistry and Group B was composed of students of all other faculties. A questionnaire was used. MAIN RESULTS: The mean age was 21.0+/-3.0 and 58% were females. The aim of antibiotic use for common cold was 83.1% (P>0.05) and, to decrease fever was 32.1% (P<0.05) in both Groups. In Group A 36.1% and in Group B 44.9% of the respondents started antibiotics by themselves when they were ill (P>0.05) although 89.1% of both groups agree that antibiotics should be started with doctors' prescription. During their last infection in Group A 11.7% and in Group B 27.3% of the respondents used the same antibiotic as previously prescribed by their doctors and the use of antibiotics as advised by the doctors during their last infection was 50.8% in Group A and 35.3% in Group B. CONCLUSIONS: The use of antibiotics are found to be irrational among university students. National education programmes about the dangers of irrational antibiotic use and restriction of antibiotics without prescriptions should be the priority. This study indicated that knowledge regarding antibiotics cannot be evaluated alone since it did not always correlate with behaviour. PMID- 16038765 TI - Major histocompatibility complex and central nervous system involvement by paracoccidioidomycosis. AB - Paracoccidioidomycosis (PCM) is a chronic granulomatous infectious disease, whose etiologic agent is the fungus Paracoccidioides brasiliensis. The central nervous system (CNS) involvement with paracoccidioidomycosis (NPCM) occurs more frequently than has been admitted in the past. There are some major histocompatibility complex antigen association studies with systemic paracoccidioidomycosis. Some indicate a positive association with HLA antigens, but there is no study with the involvement of the CNS. To investigate why not all cases of systemic PCM show the involvement of the CNS and whether genetic factors are involved, we studied 6 patients with NPCM, from the neuroinfection outpatient clinic. The patients were typed for class I and class II antigens by a microlymphocytoxity standard test. The HLA antigen frequencies found in this study in patients with NPCM were not different from the alleles frequencies observed in the Parana population. PMID- 16038766 TI - Effect of antibiotic sub inhibitory concentration on cytolethal distending toxin production by Campylobacter jejuni. AB - OBJECTIVES: To investigate the effect of pre-exposure antibiotics on cytolethal distending toxin (CDT) production and toxigenic effect of C. jejuni. METHODS: Sonicates and filtrates were prepared from known cdt+ and cdt- isolates of C. jejuni which had been pre-exposed to varying concentrations (MIC, 1/2 MIC, 1/4 MIC, 1/8 MIC) of erythromycin and ciprofloxacin. The CDT toxigenic effect was examined using INT 407 and HeLa cells. RESULTS: A trend of increased toxigenic effect was observed with pre-exposure to antibiotics. This was more pronounced with erythromycin pre-exposure compared to ciprofloxacin. Although a trend of increasing toxigenic effect with decreasing antibiotic concentration was demonstrable, some differences were observed between isolates. In one isolate the increased toxigenic effect was statistically significant (P<0.05) at 1/4 MIC in INT 407 cells and at 1/8 MIC in HeLa cells. CONCLUSIONS: This study provides evidence of an association between CDT production by C. jejuni and pre-exposure to antibiotics. Pre-exposure to ciprofloxacin and erythromycin at concentrations below MICs could potentiate CDT activity. Further work is needed to elucidate the mechanism involved. We recommend that these antibiotics be used in the treatment of C. jejuni enteritis only when strongly indicated and with careful monitoring of patients. PMID- 16038767 TI - Extracellular proteases of Acanthamoeba castellanii (encephalitis isolate belonging to T1 genotype) contribute to increased permeability in an in vitro model of the human blood-brain barrier. AB - OBJECTIVES: Granulomatous amoebic encephalitis (GAE) is a serious human infection with fatal consequences, however, the pathogenic mechanisms associated with this disease remain unclear. Several lines of evidence suggest that haematogenous spread is a prerequisite for Acanthamoeba encephalitis but it is not clear how circulating amoebae cross the blood-brain barrier to gain entry into the central nervous system. Objectives of this study were to determine the effects of Acanthamoeba on the permeability of an in vitro blood-brain barrier model and factors contributing to these changes. METHODS: Using human brain microvascular endothelial cells, an in vitro blood-brain barrier model was constructed in 24 well Transwell plates. Acanthamoeba (GAE isolate belonging to T1 genotype) or its conditioned media were used to determine permeability changes. Zymography assays were performed to characterise Acanthamoeba proteases. In addition, the ability of Acanthamoeba to bind brain microvascular endothelial cells was determined using adhesion assays. RESULTS: We observed that Acanthamoeba produced an increase of more than 45% in the blood-brain barrier permeability. Acanthamoeba conditioned media exhibited similar effects indicating Acanthamoeba-mediated blood-brain barrier permeability is contact-independent. Prior treatment of conditioned media with phenylmethyl sulfonyl fluoride (PMSF, serine protease inhibitor), abolished permeability changes indicating the role of serine proteases. Of interest, methyl alpha-d-mannopyranoside inhibited Acanthamoeba binding to human brain microvascular endothelial cells but had no effect on Acanthamoeba-mediated blood-brain barrier permeability. Zymography assays revealed that Acanthamoeba produced two major proteases, one of which was inhibited by PMSF (serine protease inhibitor) and the second with 1,10 phenanthroline (metalloprotease inhibitor). CONCLUSIONS: We have for the first time shown that Acanthamoeba produces human brain microvascular endothelial cells permeability, which can be blocked by PMSF. A metalloprotease of approx. molecular weight of 150 kDa is produced by A. castellanii (GAE isolate belonging to T1 genotype) and its role in the disease is suggested. PMID- 16038768 TI - Immunomodulatory effect of Canova medication on experimental Leishmania amazonensis infection. AB - This study investigates the action of Canova medication (CM) on experimental infection by Leishmania (Leishmania) amazonensis, utilizing in vitro and in vivo assays. For the in vitro tests, Balb/c mouse peritoneal macrophages (5x10(5) cells in 500 microl of culture medium, supplemented with 10% fetal calf serum, penicillin (100 U/ml) and streptomycin (0.1 mg/ml) (were distributed in 24-well plates and CM was added at concentrations of 20 or 40%. Twenty-four hours later, the macrophages were infected with Leishmania amastigotes in culture medium. The effect of CM on macrophages leishmanicidal activity in 24 and 48 h cultures was evaluated by determining infection index and measuring nitric oxide (NO) production. The in vivo tests were performed in mice infected with 10(7)L. (L.) amazonensis promastigotes injected in to the right hind footpad (25 microl in phosphate buffered saline). The progression of the lesions was examined over a 9 week period by measuring footpad swelling, and the parasite load in regional lymph nodes and spleen. The in vitro results showed that at 40% CM reduced the infection index, and induced NO production in the elicited macrophages, which suggests that the inhibitory effect on infection index may be mediated by NO. In the in vivo infection, when administered, orally or subcutaneously in mice, CM reduced infection by L. (L.) amazonensis in the paws, resulting in smaller lesions. CM treatment also decreased parasite load in the regional popliteal lymph nodes and in the spleen. These results suggest that CM modulates experimental infection by L. (L.) amazonensis, controlling infection progression and limiting dissemination. PMID- 16038770 TI - Subjective and physiological reactivity to chocolate images in high and low chocolate cravers. AB - Cue-reactivity to chocolate images was assessed using self-report and physiological measures. From a pre-screening sample of 454, young women were selected and assigned to high and low chocolate craving groups (N = 36/group). The experimental procedure consisted in the elicitation and measurement of the cardiac defense and startle reflexes while viewing chocolate and standard affective images selected from the International Affective Picture System. In response to chocolate images, high cravers reported more pleasure and arousal but less control than low cravers. In high cravers, viewing chocolate images inhibited the cardiac defense but potentiated the startle reflex, as compared to low cravers. The results confirmed at the physiological level that the motivational state that underlies the experience of chocolate craving include both appetitive (inhibition of the defense reflex) and aversive (potentiation of the startle response) components. The findings supported a motivational conflict theory of chocolate craving. PMID- 16038769 TI - Perceptual speed does not cause intelligence, and intelligence does not cause perceptual speed. AB - There is ongoing debate whether the efficiency of local cognitive processes leads to global cognitive ability or whether global ability feeds the efficiency of basic processes. A prominent example is the well-replicated association between inspection time (IT), a measure of perceptual discrimination speed, and intelligence (IQ), where it is not known whether increased speed is a cause or consequence of high IQ. We investigated the direction of causation between IT and IQ in 2012 genetically related subjects from Australia and The Netherlands. Models in which the reliable variance of each observed variable was specified as a latent trait showed IT correlations of -0.44 and -0.33 with respective Performance and Verbal IQ; heritabilities were 57% (IT), 83% (PIQ) and 77% (VIQ). Directional causation models provided poor fits to the data, with covariation best explained by pleiotropic genes (influencing variation in both IT and IQ). This finding of a common genetic factor provides a better target for identifying genes involved in cognition than genes which are unique to specific traits. PMID- 16038771 TI - Erotic and disgust-inducing pictures--differences in the hemodynamic responses of the brain. AB - The aim of this fMRI study was to explore brain structures that are involved in the processing of erotic and disgust-inducing pictures. The stimuli were chosen to trigger approach and withdrawal tendencies, respectively. By adding sadomasochistic (SM) scenes to the design and examining 12 subjects with and 12 subjects without sadomasochistic preferences, we introduced a picture category that induced erotic pleasure in one sample and disgust in the other sample. Since we also presented neutral pictures, all subjects viewed pictures of four different categories: neutral, disgust-inducing, erotic, and SM erotic pictures. The analysis indicated that several brain structures are commonly involved in the processing of disgust-inducing and erotic pictures (occipital cortex, hippocampus, thalamus, and the amygdala). The ventral striatum was specifically activated when subjects saw highly sexually arousing pictures. This indicates the involvement of the human reward system during the processing of visual erotica. PMID- 16038772 TI - Women's preferences for sexual dimorphism in height depend on menstrual cycle phase and expected duration of relationship. AB - Human mate preferences are related to many morphological traits, such as female waist-to-hip ratio (WHR), body mass index (BMI), male height or facial symmetry. People also vary in preferences for sexual dimorphism in stature (SDS = male height/female height) between themselves and a potential partner. Here, we demonstrate that women adjust their preference for SDS not only in relation to their own height but also in relation to (1) the phase of menstrual cycle during which their preferences were studied and (2) the sexual strategy (short- versus long-term) they were asked to choose. Taller males (larger SDS) were preferred more often when women were in the follicular (i.e. fertile) phase of their menstrual cycle and when the partners were chosen for short-term relationships. These effects were independent of woman's height. The results show that women in a potentially fertile phase of their menstrual cycle and when choosing a partner who might be less likely to invest in children select genes of taller males. PMID- 16038773 TI - A generalized physiological hyperreactivity to acute stressors in hypertensives. AB - Hypertensives have consistently been found to have a more reactive cardiovascular system than normotensives. In the present study, it was examined whether this enhanced cardiovascular stress reactivity generalizes to the hypothalamus pituitary-adrenal (HPA) axis and the immune system. Forty-two unmedicated hypertensives and 21 normotensive controls performed five passive coping and active coping stressful tasks in the laboratory. In addition to the expected greater mean diastolic blood pressure reactivity to the tasks, hypertensives exhibited enhanced (baseline corrected) task salivary cortisol and secretory immunoglobulin A (S-IgA) levels. Moreover, correlations were found between blood pressure responses and task related cortisol activity and between baseline blood pressure levels and task-induced S-IgA levels. These results indicate that hypertensives not only have a hyperreactive cardiovascular system, but also an enhanced HPA axis and immune system reactivity to stress. A central stress mechanism may be responsible for the heightened generalized stress response in hypertensives. PMID- 16038774 TI - Selective processing of pain-related word stimuli in subclinical depression as indicated by event-related brain potentials. AB - An intense discussion still exists as to whether pain and depression are causally related or independent of each other. To investigate processing of pain-related word stimuli in subclinically depressed individuals, we designed an event-related potentials study in a group control design. Pain words and neutral words were presented to 16 subclinically depressed and 16 control participants. Behavioral and electrophysiological measures were taken during lexical decision and recognition tasks. Depressive compared to control participants showed enhanced P300 amplitudes at parietal electrodes triggered by pain-related words during the lexical decision task, which presumably is a sign of enhanced recollection processes for these word stimuli. In line with these electrophysiological findings, depressed participants also tended to better recall the pain-related words in the later recognition task than control participants. We conclude that subclinically depressed individuals selectively process pain-related stimuli, and this processing bias could enhance their vulnerability to develop pain symptoms. PMID- 16038775 TI - Heart rate variability with repetitive exposure to music. AB - Previous studies of physiological responses to music showed inconsistent results, which might be attributable to methodological differences. Heart rate variability has been used to assess activation of the sympathetic and the parasympathetic nervous systems. The present study aimed to examine heart rate variability with repetitive exposure to sedative or excitative music. The participants were 13 undergraduate or graduate students who were each exposed to three conditions sedative music (SM), excitative music (EM), and no music (NM) on different days. Each participant underwent four sessions of one condition in a day. Sedative music and no music each induced both high relaxation and low tension subjectively. However, excitative music decreased perceived tension and increased perceived relaxation as the number of sessions increased. The low-frequency (LF) component of heart rate variability (HRV) and the LF/HF (high-frequency) ratio increased during SM and EM sessions but decreased during NM sessions. The HF component of HRV during SM was higher than that during EM but the same as that during NM. These findings suggest that excitative music decreased the activation of the parasympathetic nervous system. PMID- 16038776 TI - The gene trap resource: a treasure trove for hemopoiesis research. AB - The laboratory mouse is an invaluable tool for functional gene discovery because of its genetic malleability and a biological similarity to human systems that facilitates identification of human models of disease. A number of mutagenic technologies are being used to elucidate gene function in the mouse. Gene trapping is an insertional mutagenesis strategy that is being undertaken by multiple research groups, both academic and private, in an effort to introduce mutations across the mouse genome. Large-scale, publicly funded gene trap programs have been initiated in several countries with the International Gene Trap Consortium coordinating certain efforts and resources. We outline the methodology of mammalian gene trapping and how it can be used to identify genes expressed in both primitive and definitive blood cells and to discover hemopoietic regulator genes. Mouse mutants with hematopoietic phenotypes derived using gene trapping are described. The efforts of the large-scale gene trapping consortia have now led to the availability of libraries of mutagenized ES cell clones. The identity of the trapped locus in each of these clones can be identified by sequence-based searching via the world wide web. This resource provides an extraordinary tool for all researchers wishing to use mouse genetics to understand gene function. PMID- 16038777 TI - Pivotal role of peroxisome proliferator-activated receptor gamma (PPARgamma) in regulation of erythroid progenitor cell proliferation and differentiation. AB - OBJECTIVE: The aim of this study was to reveal the role of peroxisome proliferator-activated receptor gamma (PPARgamma) in erythropoiesis. METHODS: The effects of PPARgamma ligands on cellular proliferation and differentiation were investigated in erythroid colony-forming cells (ECFCs) purified from human peripheral blood. RESULTS: RT-PCR analysis revealed that PPARgamma mRNA is expressed in ECFCs. Synthetic PPARgamma ligands, troglitazone or pioglitazone, suppressed cellular proliferation without inducing apoptosis and delayed maturation of ECFCs, as determined by flow cytometry. The delay in erythroid maturation by troglitazone was confirmed by the down-regulation of gamma-globin, beta-globin and GATA-1 mRNA, and the maintenance of GATA-2 mRNA. CONCLUSIONS: Our results suggest that PPARgamma modulates the differentiation process of erythroid progenitor cells, and plays a crucial role in regulating the balance of hematopoiesis. PMID- 16038778 TI - Adenoviral-mediated transfer of TGF-beta1 but not IGF-1 induces chondrogenic differentiation of human mesenchymal stem cells in pellet cultures. AB - OBJECTIVE: The objective of the present study was to investigate the potential of application of growth factor genes to induce chondrogenic differentiation of human-derived mesenchymal stem cells (MSCs). The growth factor genes evaluated in the present study were transforming growth factor 1 (TGF-beta1) and insulin-like growth factor 1 (IGF-1). METHODS: Human MSCs were transduced with the adenoviral vectors carrying either TGF-beta1 or IGF-1 (AdTGF-beta1 and AdIGF-1 respectively) or a combination of both growth factor genes at different multiplicities of infection (MOI) and were then made into pellets. Pellets were also made from nontransduced cells and maintained in culture medium supplemented with 10 ng/mL of TGF-beta1. At specified time points, histological analysis, cartilage matrix gene expression, and immunofluorescence were performed to determine the extent of chondrogenic differentiation. RESULTS: MSCs transduced with the AdTGF-beta1 demonstrated robust chondrogenic differentiation, while those made from AdIGF-1 did not. AdTGF-beta1 pellets demonstrated aggrecan gene expression as early as day 3 of pellet culture, while type II collagen gene expression was detected by day 10 of culture. The AdIGF-1, alone or in combination with TGF-beta1 pellets, did not show any type II collagen gene expression at any time point. By immunofluoresecence, type X collagen was distributed throughout the matrix in TGF beta1 protein pellets while the growth factor gene pellets displayed scant staining. CONCLUSION: The results suggest that sustained administration of TGF beta1 may be more effective in suppressing terminal differentiation than intermittent dosing and thus effective for cartilage repair. PMID- 16038779 TI - Long-term persistence of a nonintegrated lentiviral vector in mouse hematopoietic stem cells. AB - OBJECTIVE: Lentiviral transduction is an established method for efficiently modifying the gene expression program of primary cells, but the ability of the introduced construct to persist as an episome has not been well studied. MATERIAL AND METHODS: Here we investigated this issue in lethally irradiated female mice injected with 300 or 3000 doubly sorted male lin(neg), Sca-1(high), c-kit(high), Thy-1.1(low) mouse bone marrow cells that had been exposed in vitro to self inactivating lentivirus vector encoding a green fluorescence protein (GFP) cDNA. Seven to sixteen months later, bone marrow cells from primary mice were injected into secondary female recipients and another 8 months later into tertiary female recipients. Integration study was performed on individual spleen colonies by Southern blot analysis. Inverse polymerase chain reaction (PCR) and sequence of amplified vector-derived DNA was used to verify Southern blot results. RESULTS: Spleen colony-forming cell study revealed that a small fraction of the spleen colonies contained integrated provirus as shown by Southern blot analysis. Unexpectedly, many spleen colonies were found to contain a nonintegrated episomal form of the provirus, which was confirmed by an inverse PCR analysis. In some of the spleen colonies containing only the episomal form, GFP-expressing cells were also detected. Lentiviral sequences were present in hematopoietic tissues of primary mice but not in other tissues. CONCLUSIONS: These results demonstrate that lentiviral vectors produce episomal circles in hematopoietic stem cells that can be transferred through many cell generations and expressed in their progeny. PMID- 16038780 TI - Identification of progression markers in B-CLL by gene expression profiling. AB - OBJECTIVE: B-cell chronic lymphocytic leukemia is a heterogeneous disease with a pronounced variation in the clinical course. With the purpose of identifying genes that could be related to disease progression, we have performed gene expression profiling on B-CLL patients with an indolent disease and patients with a progressive disease with need for therapy. MATERIALS AND METHODS: we applied the Affymetrix GeneChip technique to 11 B-CLL patients with stable and 10 patients with clinically progressive disease. Supervised and unsupervised clustering methods with different algorithms were used to identify genes that tend to give a distinction between stable and progressive disease. RESULTS: The supervised learning procedures identified groups of genes with a combined power to discriminate samples from progressive and stable disease with 70-90% accuracy. The gene for protein phosphatase 2 regulatory subunit B' (B56) gamma isoform (PPP2R5C) and the gene for retinoblastoma-like 2 (p130) (RBL2) were included among the best discriminators; both genes were downregulated in progressive as compared to stable B-CLL. In a hierarchical clustering analysis based on gene expression pattern three clinical subcategories could be identified: one with a more severe clinical outcome, a second one with good prognosis, and a third one that was intermediate between the other two groups. CONCLUSIONS: Our application of microarray analysis on a clinically well defined material has identified a number of genes with combined expression patterns related to stable or progressive disease in general. Unsupervised clustering suggested the existence of subclasses of samples in the progressive group that may be identifiable through gene expression patterns. PMID- 16038781 TI - Numbers of Foxp3-expressing CD4+CD25high T cells do not correlate with the establishment of long-term tolerance after allogeneic stem cell transplantation. AB - OBJECTIVE: Regulatory CD4 T cells that express high levels of CD25 play a vital role in the maintenance of tolerance to self antigens and are required for the induction of nonresponsiveness to alloantigens. The long-term CD4+CD25high T-cell reconstitution after allogeneic stem cell transplantation is unknown. Here, we evaluated whether recovery of this T-cell subset might be linked to the establishment of full donor/recipient tolerance. METHODS: The frequency of CD4+CD25high T cells was determined by Fluorescence Activated Cell Sorter (FACS) analysis in 31 patients, with a mean follow-up of more than 31 months posttransplant. The expression levels of Foxp3 mRNA were assessed by quantitative real-time polymerase chain reaction (RT-PCR). RESULTS: Patients with or without graft-versus-host disease (GvHD) had significant and persistent CD4 T-cell lymphopenia. The relative frequency of CD25high cells and the expression levels of FoxP3 mRNA within this subset were similar between all patients and healthy controls. No significant difference was found in the number of Foxp3-expressing CD4+CD25high T cells in patients with or without GvHD. Finally, younger age and absence of previous GvHD were significantly linked to CD4+CD25high T-cell recovery. CONCLUSION: The low number of Foxp3-expressing CD4+CD25high T cells in grafted patients is not a specific default of this compartment but a consequence of global CD4 T-cell lymphopenia after allogeneic stem cell transplantation. Moreover, levels of Foxp3 mRNA in the CD25+ T-cell compartment do not allow predicting the development of GvHD in the long term. PMID- 16038782 TI - Defective stromal cell function in a mouse model of infusion-induced bone marrow failure. AB - OBJECTIVE: To study bone marrow (BM) stromal damage in a mouse model of infusion induced BM failure. MATERIALS AND METHODS: Sublethally irradiated CByB6F1 mice were infused with 5 x 10(6) C57BL/6 (B6) lymph node (LN) cells. Recipient BM cells were taken at 3, 7, 10, and 14 days following LN infusion and were cultured in vitro in alpha-modified Eagle media for 2-3 weeks. Peripheral blood and was analyzed by complete blood counts while BM lymphocyte infiltration/expansion was analyzed by flow cytometry. Marrow cells from affected and control mice were mixed and cultured in vitro to test nonspecific stromal damage. RESULTS: Donor lymphocytes infiltrated host BM within 3-7 days and expanded significantly between 7 and 10 days, concurrent with the development of leukopenia, thrombocytopenia, and marrow hypoplasia. BM cells from mice at 7, 10, and 14 days after B6-LN cell infusion were progressively defective in forming stromal feeder layers. A 1:1 mixture of BM cells from affected CByB6F1 mice and normal B6 mice failed to form an effective stromal feeder layer that could support cobblestone colony formation, indicating that lymphocytes in the BM of affected CByB6F1 mice were able to damage stromal cells in the normal B6 BM. CONCLUSION: Activated lymphocytes destroy both hematopoietic and stromal cells as innocent bystanders in the infusion-induced BM failure model. PMID- 16038783 TI - No evidence of plasticity in hair follicles of recipients after allogeneic hematopoietic stem cell transplantation. AB - Here we show in a prospective quantitative study of 115 patients after allogeneic hematopoietic stem cell transplantation that hair follicles remain exclusively of recipient type despite full whole blood donor-type chimerism. Our data indicate that unmanipulated hematopoietic donor stem cells do not contribute directly to reconstitution even in an organ at highest need for repair. PMID- 16038784 TI - A comparison of postengraftment infectious morbidity and mortality after allogeneic partially T cell-depleted peripheral blood progenitor cell transplantation versus T cell-depleted bone marrow transplantation. AB - OBJECTIVE: Postengraftment infections are a major cause of transplant-related morbidity and mortality following allogeneic hematopoietic stem cell transplantation (allo-SCT). Allogeneic peripheral blood progenitor cell transplantation (PBPCT) is associated with faster hematopoietic recovery compared to bone marrow transplantation (BMT) and unmanipulated PBPCT may be associated with fewer postengraftment infections. We set out to evaluate and compare the incidence, cause, and outcome of postengraftment infections following HLA identical sibling T cell-depleted PBPCT vs T cell-depleted BMT between days 30 and 365 posttransplant. PATIENTS: Forty recipients of peripheral blood progenitor cells (PBPC) and 47 recipients of bone marrow (BM) were included. The two groups of patients were comparable with respect to their baseline characteristics. RESULTS: PBPC grafts contained significantly more CD34+ cells and PBPCT was associated with significantly faster neutrophil and lymphocyte recovery as compared to BMT. PBPC recipients experienced more chronic graft-vs-host disease (GVHD; 55% vs 34%; p=0.02). The number of definite and clinical infections per 100 patient days was comparable between recipients of PBPC and BM with similar contribution of causative microorganisms. At one year post SCT, 68% of PBPC recipients had experienced at least one CTC grade 3-4 infection vs 65% of BM recipients. Treatment-related mortality at one year from transplantation was 34% after PBPCT vs 30% after BMT, and no difference in infection-related mortality was observed. CONCLUSION: Postengraftment infectious morbidity and mortality were comparable between recipients of PBPC and BM despite a higher CD34+ cell content of PBPC grafts and faster lymphocyte recovery after PBPCT, which may in part be explained by the higher incidence of chronic GVHD. PMID- 16038785 TI - Reduced-intensity conditioning containing low-dose alemtuzumab before allogeneic peripheral blood stem cell transplantation: graft-versus-host disease is decreased but T-cell reconstitution is delayed. AB - OBJECTIVE: In vivo administration of alemtuzumab (an anti-CD52 antibody) is effective to decrease the incidence of graft-versus-host disease (GVHD) after allogeneic stem cell transplantation (allo-SCT). However, posttransplant immune reconstitution is impaired, increasing the infection risk. We investigated the effect of in vivo administration of a low-dose alemtuzumab on GVHD prevention and kinetics of immune reconstitution. PATIENTS AND METHODS: Twenty-seven patients entered a pilot study employing reduced-intensity conditioning and low-dose alemtuzumab (15 or 7.5 mg/m2) before peripheral blood allo-SCT from human leukocyte antigen-identical or one antigen-mismatched sibling donors. All lymphoid subsets were longitudinally studied at 1-3, 6, 9, 12 months after transplantation. T-cell receptor (TCR) spectratyping and T-cell receptor excision circles (TRECs) were also analyzed at various time points after allo-SCT. RESULTS: All patients but one were engrafted. The probability of nonrelapse mortality at 100 days and 1 year were 7 and 11%, respectively; the overall survival at 2 years was 77%. The cumulative incidence of grade II-IV acute GVHD at day 100 was 11%. The overall incidence of chronic GVHD was 28%. The median time to achieve more than 200 CD4+/microL and 500 CD8+/microL were 6 and 9 months, respectively. Natural killer cells remained between the value of 300/microL and 500/microL throughout the period of follow-up whereas the median time to reach CD19+ blood concentrations of >200 cells/microL was 9 months. The normalization of TCR repertoire and increase of TREC counts began at 6 months after allo-SCT. CONCLUSION: We have shown that low-dose alemtuzumab is effective for GVHD prevention, but its use still impairs the immune reconstitution. PMID- 16038786 TI - Human mesenchymal stem cells require monocyte-mediated activation to suppress alloreactive T cells. AB - Human bone marrow-derived mesenchymal cells (MSCs) are precursors of nonhematopoietic mesenchymal cells of the bone marrow microenvironment. MSCs were shown to inhibit alloreactive T lymphocytes, but the mechanism and mediators of this effect are not fully understood. Here we describe a novel interaction between blood monocytes and bone marrow-derived, culture-expanded MSCs, which results in inhibition of T-lymphocyte activation. We found that CD14+ monocytes from blood activate MSCs to secrete inhibitory molecules that lead to inhibition of alloreactive T cells. This cellular communication is not contact-dependent, but rather is mediated by soluble factors that include interleukin (IL)-1beta. MSC-mediated inhibition of alloreactive T lymphocytes is associated with downregulation of activation markers CD25, CD38, and CD69 detected both in CD4+ and CD8+ T lymphocytes. The cytokines secreted by MSCs that mediate T-cell inhibition include transforming growth factor-beta1, but not IL-10. The interaction between blood monocytes and the MSCs represents a unique immune regulatory paradigm that can potentially be exploited in clinic. PMID- 16038787 TI - Posttransplant hematopoiesis in patients undergoing sibling allogeneic stem cell transplantation reflects that of their respective donors although with a lower functional capability. AB - We tested the principle of whether patient long-term hematopoiesis following allogeneic stem cell transplantation (allo-SCT) reflects the characteristics of the hematopoiesis of their respective donor. For this purpose, we analyzed bone marrow (BM) hematopoiesis using long-term cultures (LTC), delta assays, and clonogeneic assays as well as CD34+ cells and their subsets by flow cytometry in a series of 37 patients undergoing allo-SCT, and we compared it to that of their respective human leukocyte antigen-matched sibling donors in a paired study performed more than 1 year after the transplant procedure. Interestingly, the main factor that influenced post-allo-SCT BM hematopoiesis in the long term was donor hematopoiesis. Nevertheless, compared to their respective donors, patients exhibited a significantly lower number of colony-forming units granulomonocytic, burst-forming units erythroid, and immature progenitors (CD34++/CD38dim/CD90+/CD133+ cells, LTC-initiating cells, and colonies generated in the delta assay). Moreover, BM stromal function was diminished in patients undergoing allo-SCT compared to their donors. In addition, the presence of chronic graft-versus-host disease under immunosuppressive treatment also conditioned an impaired hematopoietic function. In summary, our study shows that BM hematopoiesis evaluated more than 1 year after an allo-SCT mainly reproduces that of their respective donors, although with a significantly decreased in vitro activity. PMID- 16038788 TI - Mast cell activation is differentially affected by heat shock. AB - OBJECTIVE: Mast cells play pivotal roles in immediate-type and inflammatory allergic and nonallergic reactions. Cross-linking of the high-affinity receptor for IgE (FcepsilonRI) on mast cells activates a signaling pathway leading to Ca2+ mobilization and is followed by degranulation and the release of histamine and other preformed mediators, as well as de novo synthesis of arachidonic acid metabolites. In a previous study, we have demonstrated that heat shock activates heat shock transcription factor-1 (HSF-1), induces heat shock protein 70 (HSP70), and suppresses cytokine production in bone marrow-derived mast cells (BMMC). In this study, we further investigated the effects of heat shock on the activation of mast cells and the release of mast cell mediators. METHODS: In mouse mast cells, derived from a culture of bone marrow cells of male BALB/cBy and null HSF 1(-/-)mice, responsiveness to heat shock was monitored by measuring beta hexosaminidase and leukotriene C4 (LTC4) release. RESULTS: Using BMMC, we found that heat shock inhibits degranulation of BMMC without affecting leukotriene production. To further elucidate the mechanism of suppression of degranulation, we studied the effects of heat shock on the regulation of signal transduction in more detail. We found that heat shock inhibits calcium mobilization and tyrosine phosphorylation of Syk and SHIP upon IgE receptor activation, but increases the phosphorylation of SHP-1 and -2. Moreover, our results revealed that suppression of tyrosine phosphorylation of Syk and SHIP coincided with an increased tyrosine phosphatase activity. CONCLUSION: The inhibitory action of heat shock toward mast cell degranulation is likely due to shifting the balance between kinase and phosphatase activity. PMID- 16038789 TI - In silico analysis of angiogenesis associated gene expression identifies angiogenic stage related profiles. AB - In vitro models have been extensively used to map gene expression in ECs but few studies have used cells from in vivo sources directly. Here, we compare different gene expression surveys on both cultured and fresh tissue derived ECs, and it emerges that gene expression profiles can be paralleled with the angiogenic stage of the cells. ECs stimulated with different growth factors in monolayer cultures exhibit gene expression profiles indicative of an active proliferative state, whereas gene expression in tube forming cells in vitro involves genes implicated in cell adhesion processes. Genes overexpressed in tumor ECs are biased towards extracellular matrix remodeling, a late event in angiogenesis. The elucidation of gene expression profiles under these different conditions will contribute to a better understanding of the molecular mechanisms during angiogenesis in both pathological and physiological circumstances and will have implications for the development of angiogenesis interfering treatment strategies. PMID- 16038790 TI - Transcriptional profiles of regulatory and virulence factors of Staphylococcus aureus of bovine origin: oxygen impact and strain-to-strain variations. AB - Staphylococcus aureus is responsible for a large panel of infections in humans and animals. In cows, S. aureus provokes chronic intramammary infections. Little information is available about the regulation of virulence factors in bovine isolates. Moreover, oxygenation, which is low in an inflamed mammary gland, could play an important role during the infectious process. We investigated the impact of oxygen on regulatory and virulence factors transcription for three S. aureus bovine isolates cultivated in CYPG medium into a fermentor under moderate oxygenation or low oxygenation. A selective panel of regulatory factors and virulence factors was studied through their mRNA profiles by real-time PCR according to growth phases and oxygenation. RNAIII, rot and sarR genes, for the regulatory factors, and asp23 and cflA genes, for the virulence factors, were strongly expressed, whatever the oxygenation and the strains. Under low oxygenation, whatever the strain, an enhanced expression of srr, clfA and spa genes was detected. Some regulators such as sae, sarA and sigB were differentially transcribed according to the strain and the oxygenation condition. This study sustains the complexity of S. aureus genes global regulation and suggests the coexistence of different pathways that can be activated depending on the strain and the oxygen availability. PMID- 16038791 TI - Comparison between spoligotyping and IS6110 restriction fragment length polymorphisms in molecular genotyping analysis of Mycobacterium tuberculosis strains. AB - Spoligotyping was compared with RFLP fingerprinting analysis in the identification of Mycobacterium tuberculosis strains. Spoligotyping sensitivity was 97.6% with a specificity of 47%. The global probability for two strains clustered with spoligotyping to be clustered also with RFLP analysis was 33%; the probability for two strains clustered with RFLP analysis to be clustered also with spoligotyping analysis was 95%. However, comparing the two methods in five outbreak episodes, full concordance was evidenced between spoligotyping and RFLP. Moreover, we evaluated the presence of our 17 largest spoligotyping clusters in spoligotyping databases from Caribbean countries, London and Cuba. Only five out of 17 patterns were present in all the cohorts. The conditional probability comparing spoligotyping and RFLP methods related to these patterns resulted in very low concordance (range from 2 to 38%). In conclusion, we confirm that spoligotyping when used alone overestimates the number of recent transmission and does not represent a suitable method for wide clinical practice application. However, it allows to get a first good picture of strain identity in a new setting and in more localized or confined settings, the probability of reaching the same result compared to RFLP was 100% confirming the usefulness of spoligotyping in the management of epidemic events, especially in hospitals, prisons and close communities. PMID- 16038792 TI - Molecular characterization of Thelazia lacrymalis (Nematoda, Spirurida) affecting equids: a tool for vector identification. AB - Equine thelaziosis caused by the eyeworm Thelazia lacrymalis is a parasitic disease transmitted by muscid flies. Although equine thelaziosis is known to have worldwide distribution, information on the epidemiology and presence of the intermediate hosts of T. lacrymalis is lacking. In the present work, a PCR-RFLP based assay on the first and/or second internal transcribed spacer (ITS1 and ITS2) of ribosomal DNA was developed for the detection of T. lacrymalis DNA in its putative vector(s). The sensitivity of the technique was also assessed. The restriction patterns obtained readily differentiated T. lacrymalis from four species of Musca (Diptera, Muscidae) (i.e. Musca autumnalis, Musca domestica, Musca larvipara and Musca osiris), which are potential vectors of equine eyeworms. The molecular assay presented herein is a useful tool to identify the intermediate host(s) of T. lacrymalis in natural conditions and to study its/their ecology and epidemiology. PMID- 16038793 TI - Copro-PCR based detection of Schistosoma eggs using mitochondrial DNA markers. AB - We report on a sensitive, specific and easy to interpret PCR based diagnostic tool for the detection of Schistosoma japonicum eggs in the faeces of infected mammalian hosts. Primer pairs were designed to amplify regions of the mitochondrial DNA of the parasite. The specificity of the PCR primers was tested using either faecal samples from non-infected hosts or hosts infected with the related schistosome species S. mansoni. Sensitivity was investigated in a study, which differentiated the presence or absence of eggs in faecal samples. PCR results were correlated with analysis of the samples by microscopy. PCR analysis provided a level of sensitivity of 87.7%, while specificity was 100%. The PCR based assay could detect mitochondrial DNA from as little as 0.3 of a single egg. The overall detection threshold of the PCR test was >or=60 eggs per gram of faeces. Advantages of this technique include the ability to scale-up screening and the reproducibility and simplicity of interpretation of results compared with standard microscopic methods. PMID- 16038794 TI - Heterologous regulation of chemokine receptor signaling by the lipid phosphatase SHIP in lymphocytes. AB - The SH2 domain-containing inositol polyphosphate 5-phosphatase (SHIP) is known to play an important role in the negative regulation by FcgammaRIIB of PI3K dependent signaling cascades activated by the B cell antigen receptor (BCR) as well as several tyrosine-kinase coupled cytokine receptors. However, to date the role of SHIP in the regulation of PI3K-dependent signals elicited by G-protein coupled receptors (GPCR) such as chemokine receptors has not been investigated. In this study, we report that ligation of the G-protein-coupled chemokine receptor CXCR4 by SDF-1/CXCL12 has no effect on the tyrosine phosphorylation of SHIP in the murine B cell lymphoma A20. However, co-ligation of the B cell antigen receptor and FcgammaRIIB inhibits the PI3K-dependent phosphorylation of PKB and ERK1/2 in response to CXCL12. We have also utilised a constitutively active membrane-localised SHIP mutant expressed in the Jurkat leukaemic T cell line (which do not normally express SHIP), in order to investigate the effect of this mutant on CXCL12 stimulated PI3K-dependent signaling events. Experiments have revealed that CXCL12-mediated PKB phosphorylation, chemotaxis and lipid accumulation are inhibited in the presence of this SHIP mutant. Thus, it appears that heterologous activation of SHIP by non-G-protein-coupled receptor-mediated routes can impinge on PI3K-dependent signaling pathways activated by independently ligated G-protein-coupled chemokine receptors. PMID- 16038795 TI - Basal and induced sphingosine kinase 1 activity in A549 carcinoma cells: function in cell survival and IL-1beta and TNF-alpha induced production of inflammatory mediators. AB - Sphingosine-1-phosphate, a lipid mediator produced by sphingosine kinases, regulates diverse cellular processes, ranging from cell growth and survival to effector functions, such as proinflammatory mediator synthesis. Using human A549 epithelial lung carcinoma cells as a model system, we observed transient upregulation of sphingosine kinase type 1 (SPHK1) enzyme activity upon stimulation with both TNF-alpha or IL-1beta. This transient activation of SPHK1 was found to be required for cytokine-induced COX-2 transcription and PGE2 production, since not only specific siRNA (abolishing both basal and induced SPHK1 enzyme activity), but also a dominant-negative SPHK1 mutant (suppressing induced SPHK1 activity only) both reduced COX-2 and PGE2. Furthermore, TNF-alpha- or IL-1beta-induced transcription of selected cytokines, chemokines, and adhesion molecules (IL-6, RANTES, MCP-1, and VCAM-1) was found to require SPHK1 activation. Suppression of SPHK1 activation led to reduction of cytokine-induced IkappaBalpha phosphorylation and consequently diminished NFkappaB activity due to reduced nuclear translocation of RelA (p65), explaining the dependence of inflammatory mediator production on SPHK1 activation. Inhibition of basal SPHK1 activity by N,N-dimethylsphingosine or by downregulation of its expression using siRNA induced spontaneous apoptosis in A549 cells, an effect that can be explained through interference with constitutive NFkappaB activity in this cell type. In contrast, expression of the dominant-negative mutant did not induce apoptosis. Taken together, these findings demonstrate a role of SPHK1 activation in proinflammatory signalling and of SPHK1 basal activity in survival of A549 lung carcinoma cells. PMID- 16038796 TI - Loss of association between activated Galpha q and Gbetagamma disrupts receptor dependent and receptor-independent signaling. AB - The G protein subunit, betagamma, plays an important role in targeting alpha subunits to the plasma membrane and is essential for binding and activation of the heterotrimer by heptahelical receptors. Mutation of residues in the N terminal alpha-helix of alpha s and alpha q that contact betagamma in the crystal structure of alpha i reduces binding between alpha and betagamma, inhibits plasma membrane targeting and palmitoylation of the alpha subunit, and results in G proteins that fail to couple receptor activation to stimulation of effector. Overexpression of betagamma can recover this loss of signaling through Gs but not Gq. In fact, a single mutation (I25A) in alpha q can block alpha q-mediated generation of inositol phosphates. Function is not recovered by betagamma overexpression nor myristoylation directed plasma membrane localization. Introduction of a Q209L activating mutation with I25A results in a constitutively active alpha q as expected, but surprisingly a R183C activating mutation does not result in constitutive activity when present with I25A. Examination of binding between alpha and betagamma via a pull down assay shows that the N-terminal betagamma-binding mutations inhibit alpha-betagamma binding significantly more than the R183C or Q209L activating mutations do. Moreover, introduction of the I25A mutation into alpha q RC disrupts co-immunoprecipitation with PLCbeta1. Taken together, results presented here suggest that alpha-betagamma binding is necessary at a point downstream from receptor activation of the heterotrimeric G protein for signal transduction by alpha q. PMID- 16038797 TI - Phosphorylation of the retinoid x receptor at the omega loop, modulates the expression of retinoic-acid-target genes with a promoter context specificity. AB - The retinoid response is mediated by two classes of nuclear receptors, the retinoic acid receptors (RARalpha, beta, and gamma) and the retinoid X receptors (RXRalpha, beta, and gamma) which act as ligand-dependent heterodimeric RAR/RXR transcription activators. Like most transcription factors, RARs and RXRs are regulated by phosphorylation processes. Here, we report that stress agents induce RXRalpha phosphorylation, subsequently to the activation of the stress-activated protein kinases cascade (JNKs). This phosphorylation process concerns three residues located in the N-terminal AF-1 domain of RXRalpha and one located in the omega loop of the Ligand Binding Domain. To decipher how stress-induced RXRalpha phosphorylation influences the transcription of RA-target genes, we used a ribotoxic stress agent, anisomycin, which activates signaling kinases without promoting DNA or protein damages, at subinhibitory concentrations. Taking advantage of vectors expressing recombinant RXRalpha mutated at its phosphorylation sites and of F9 cell lines re-expressing the same RXRalpha mutants in an RXRalpha null background, we provide evidence that stress signaling modulates RAR/RXRalpha-mediated transcription, through the phosphorylation of RXRalpha at the residue located in the Omega loop, in a promoter context dependent manner. PMID- 16038799 TI - Beta-arrestin2 enhances beta2-adrenergic receptor-mediated nuclear translocation of ERK. AB - Beta-arrestin mediates desensitization and internalization of beta-adrenergic receptors (betaARs), but also acts as a scaffold protein in extracellular signal regulated kinase (ERK) cascade. Thus, we have examined the role of beta-arrestin2 in the betaAR-mediated ERK signaling pathways. Isoproterenol stimulation equally activated cytoplasmic and nuclear ERK in COS-7 cells expressing beta1AR or beta2AR. However, the activity of nuclear ERK was enhanced by co-expression of beta-arrestin2 in beta2AR-but not beta1AR-expressing cells. Pertussis toxin treatment and blockade of Gbetagamma action inhibited beta-arrestin2-enhanced nuclear activation of ERK, suggesting that beta-arrestin2 promotes nuclear ERK localization in a Gbetagamma dependent mechanism upon receptor stimulation. beta2AR containing the carboxyl terminal region of beta1AR lost the beta arrestin2-promoted nuclear translocation. As the carboxyl terminal region is important for beta-arrestin binding, these results demonstrate that recruitment of beta-arrestin2 to carboxyl terminal region of beta2AR is important for ERK localization to the nucleus. PMID- 16038798 TI - Galectin-1 is an inductor of pancreatic stellate cell activation. AB - Pancreatic stellate cells (PSCs) play a key role in the development of pancreatic fibrosis, a pathological feature of chronic pancreatitis and pancreatic cancer. Here, we show that activation of rat PSCs in vitro is associated with increased expression of galectin-1 (gal-1) and that gal-1 modulates PSC function. Expression of the lectin was stimulated by fetal calf serum and platelet-derived growth factor. PSCs exposed to exogenous gal-1 proliferated at a higher rate and synthesised more collagen than controls. Gal-1-dependent collagen synthesis was blocked by lactose but not by cellobiose, suggesting that gal-1 acts on PSCs through targeting beta-galactoside-containing glycoconjugates. Analysis of gal-1 signalling in PSCs revealed an activation of the extracellular signal-regulated kinases 1 and 2 and enhanced DNA binding of AP-1 transcription factors. Together, our data implicate gal-1 in PSC activation and suggest further studies to analyse the role of endogenous lectins in the development of pancreatic fibrosis in vivo. PMID- 16038800 TI - Conditional expression of MAP kinase phosphatase-2 protects against genotoxic stress-induced apoptosis by binding and selective dephosphorylation of nuclear activated c-jun N-terminal kinase. AB - MAP Kinase Phosphatase-2 (MKP-2) is a dual specific nuclear phosphatase which is selective for both ERK and JNK, MAP kinases implicated in the regulation of apoptosis in response to genotoxic stress. Here we report the conditional expression of MKP-2 in human embryonic kidney cells 293. We demonstrate that Flag WT-MKP-2 is able to rescue cells from apoptotic commitment when subjected to UV-C or cisplatin treatment. We establish that upon stimulation all three major MAP kinase families (ERK, JNK and p38 MAP kinases) are activated. However, MKP-2 is surprisingly only able to deactivate JNK in vivo. Furthermore, whilst pre treatment of cells with either the JNK inhibitor SP600125, or the MEK-1 inhibitor PD98059, also reverses UV-C and cisplatin-induced apoptosis, the anti-apoptotic effect of MKP-2 overexpression is not additive with SP600125 but is with PD098059, suggesting that MKP-2 is involved in specifically terminating JNK activity and not ERK. The inability of MKP-2 to dephosphorylate ERK in vivo is also not due to the inability of Flag-MKP-2 to bind both ERK and JNK; phosphorylated forms of each kinase are co-precipitated with both WT and CI-MKP 2. Immunofluorescence studies however demonstrate that ERK is exclusively cytosolic in origin and not translocated to the nucleus following UV-C and cisplatin treatment whilst JNK is principally nuclear. These studies demonstrate the in vivo specificity of MKP-2 for JNK and not ERK and show that nuclear targeted JNK is involved in genotoxic stress-induced apoptosis. PMID- 16038801 TI - Okadaic acid induces phosphorylation and translocation of myosin phosphatase target subunit 1 influencing myosin phosphorylation, stress fiber assembly and cell migration in HepG2 cells. AB - It was determined that the myosin phosphatase (MP) activity and content of myosin phosphatase target subunit 1 (MYPT1) were correlated in subcellular fractions of human hepatocarcinoma (HepG2) cells. In control cells MYPT1 was localized in the cytoplasm and in the nucleus, as determined by confocal microscopy. Treatment of HepG2 cells with 50 nM okadaic acid (OA), a cell-permeable phosphatase inhibitor, induced several changes: 1) a marked redistribution of MYPT1 to the plasma membrane associated with an increased level of phosphorylation of MYPT1 at Thr695. Both effects showed only a slight influence with the Rho-kinase inhibitor, Y-27632; 2) an increase in phosphorylation of MYPT1 at Thr850 associated with its accumulation in the perinuclear region and nucleus. These effects were markedly reduced by Y-27632; 3) an increased phosphorylation of the 20 kDa myosin II light chain at Ser19 associated with an increased location of myosin II at the cell center. These effects were partially counteracted by Y 27632; 4) an increase in stress fiber formation and a decrease in cell migration, both OA-induced effects were blocked by Y-27632. In HepG2 lysates, OA (5-100 nM) did not affect MP activity but inhibited PP2A activity. These results indicate that OA induces differential phosphorylation and translocation of MYPT1, dependent on PP2A and, to varying extents, on ROK. These changes are associated with an increased level of myosin II phosphorylation and attenuation of hepatic cell migration. PMID- 16038802 TI - Regulation of ASAP1 by phospholipids is dependent on the interface between the PH and Arf GAP domains. AB - ASAP1 is an Arf GAP with a PH domain immediately N-terminal to the catalytic Arf GAP domain. PH domains are thought to regulate enzymes by binding to specific phosphoinositide lipids in membranes, thereby recruiting the enzyme to a site of action. Here, we have examined the functional relationship between the PH and Arf GAP domains. We found that GAP activity requires the cognate PH domain of ASAP1, leading us to hypothesize that the Arf GAP and PH domains directly interact to form the substrate binding site. This hypothesis was supported by the combined results of protection and hydrodynamic studies. We then examined the role of the PH domain in the regulation of Arf GAP activity. The results of saturation kinetics, limited proteolysis, FRET and fluorescence spectrometry support a model in which regulation of the GAP activity of ASAP1 involves a conformational change coincident with recruitment to a membrane surface, and a second conformational change following the specific binding of phosphatidylinositol 4,5-bisphosphate. PMID- 16038804 TI - Activation and nuclear translocation of ERK1/2 by the formyl peptide receptor is regulated by G protein and is not dependent on beta-arrestin translocation or receptor endocytosis. AB - G protein-coupled receptors (GPCRs) transmit diverse cellular signals in response to a large number of stimuli such as chemoattractants, lipids, neurotransmitters, odorants and light. The classical signaling pathway is through heterotrimeric G proteins, but GPCRs can also transmit signals through mechanisms that are not dependent on G proteins. In mammalian cells, the key component for this type of signaling is the family of scaffolding molecules called beta-arrestins. They can function as scaffolds for activation of mitogen-activated protein kinases, including extracellular signal-regulated kinases 1 and 2 (ERK1/2). In this study we examined the role of G protein and beta-arrestin in formyl peptide receptor (FPR)-mediated activation of chemotaxis, receptor endocytosis and ERK1/2 activation using wild type and mutant receptors. Our findings suggest that, unlike certain other GPCRs that can activate ERK1/2 without the involvement of G protein, FPR requires signaling through a G protein-mediated pathway. Previous observations have shown that ERK1/2, activated through G protein, translocates to the nucleus where it stimulates transcription factors. In contrast, the scaffolding protein beta-arrestin retains the activated ERK1/2 in the cytoplasm to allow phosphorylation of cytoplasmic targets. Our experimental data show that both wild-type FPR and a mutant FPR, defective in beta-arrestin binding, induce nuclear translocation of activated ERK1/2 with similar ligand concentration dependence as seen for activation of cytosolic ERK1/2. We propose that FPR mediated activation of ERK1/2 takes place primarily through G protein and is physiologically important to ensure transcriptional activation of myeloid immunomodulators, such as cytokines. PMID- 16038803 TI - Grb2 negatively regulates epidermal growth factor-induced phospholipase C-gamma1 activity through the direct interaction with tyrosine-phosphorylated phospholipase C-gamma1. AB - Phospholipase C-gamma1 (PLC-gamma1) plays pivotal roles in cellular growth and proliferation. Upon the stimulation of growth factors and hormones, PLC-gamma1 is rapidly phosphorylated at three known sites; Tyr771, Tyr783 and Tyr1254 and its enzymatic activity is up-regulated. In this study, we demonstrate for the first time that Grb2, an adaptor protein, specifically interacts with tyrosine phosphorylated PLC-gamma1 at Tyr783. The association of Grb2 with PLC-gamma1 was induced by the treatment with epidermal growth factor (EGF). Replacement of Tyr783 with Phe completely blocked EGF-induced interaction of PLC-gamma1 with Grb2, indicating that tyrosine phosphorylation of PLC-gamma1 at Tyr783 is essential for the interaction with Grb2. Interestingly, the depletion of Grb2 from HEK-293 cells by RNA interference significantly enhanced increased EGF induced PLC-gamma1 enzymatic activity and mobilization of the intracellular Ca2+, while it did not affect EGF-induced tyrosine phosphorylation of PLC-gamma1. Furthermore, overexpression of Grb2 inhibited PLC-gamma1 enzymatic activity. Taken together, these results suggest Grb2, in addition to its key function in signaling through Ras, may have a negatively regulatory role on EGF-induced PLC gamma1 activation. PMID- 16038805 TI - Effect of light quality on the accumulation of photosynthetic pigments, proteins and mycosporine-like amino acids in the red alga Porphyra leucosticta (Bangiales, Rhodophyta). AB - The effect of different light qualities (white, blue, green, yellow and red light) on photosynthesis, measured as chlorophyll fluorescence, and the accumulation of photosynthetic pigments, proteins and the UV-absorbing mycosporine-like amino acids (MAAs) was studied in the red alga Porphyra leucosticta. Blue light promoted the highest accumulation of nitrogen metabolism derived compounds i.e., MAAs, phycoerythrin and proteins in previously N-starved algae after seven days culture in ammonium enriched medium. Similar results were observed in the culture under white light. In contrast, the lowest photosynthetic capacity i.e., lowest electron transport rate and lowest photosynthetic efficiency as well as the growth rate were found under blue light, while higher values were found in red and white lights. Blue light favored the accumulation of the MAAs porphyra-334, palythine and asterina-330 in P. leucosticta. However, white, green, yellow and red lights favored the accumulation of shinorine. The increase of porphyra-334, palythine and asterina-330 occurred in blue light simultaneous to a decrease in shinorine. The accumulation of MAAs and other nitrogenous compounds in P. leucosticta under blue light could not be attributed to photosynthesis and the action of a non-photosynthetic blue light photoreceptor is suggested. A non-photosynthetic photoreceptor could be also involved in the MAAs interconversion pathways in P. leucosticta. PMID- 16038806 TI - Interactions of the plant flavonoid fisetin with macromolecular targets: insights from fluorescence spectroscopic studies. AB - Fisetin (3,7,3',4'-tetrahydroxyflavone) is a bioactive plant flavonoid of immense importance as a potentially useful therapeutic drug, for various free radical mediated as well as other diseases. In a recent paper, we demonstrated the novel uses of the exquisitely sensitive intrinsic fluorescence of this compound to explore its binding characteristics in liposomal membranes [B. Sengupta, A. Banerjee, P.K. Sengupta, Investigations on the binding and antioxidant properties of the plant flavonoid fisetin in model biomembranes, FEBS Lett. 570 (2004) 77 81]. Here, we have exploited this technique to examine its interactions with relevant macromolecular targets, namely double stranded DNA (from calf thymus), and the physiologically important circulatory protein, Human Serum Albumin (HSA). In the presence of DNA dramatic changes are observed in the intrinsic fluorescence behaviour of fisetin. These, along with other relevant supporting spectroscopic data, suggest that fisetin binds intercalatively between the base pairs of DNA. From the studies on fisetin-HSA interaction, the existence of two distinct binding sites are inferred. Furthermore we present evidence for the occurrence of efficient Forster type fluorescence resonance energy transfer from tryptophan to fisetin, indicating that both binding sites of fisetin in HSA are proximal to the unique tryptophan - 214 residue present in the interdomain (between IIA and IIIA domains) loop region of the protein. PMID- 16038807 TI - Thermal stability of UV-irradiated collagen in bovine lens capsules and in bovine cornea. AB - The thermal stability of UVB irradiated collagen in bovine lens capsules and in bovine cornea has been investigated by differential scanning calorimetry (DSC). During UVB irradiation the lens capsules and cornea were immersed in water to keep the collagen in a fully hydrated condition at all times. UV irradiation induced changes in collagen which caused both stabilization and destabilization of the collagen structure. The helix-coil transition for non-irradiated collagen in cornea occurred near 66 degrees C, instead for the irradiated one for 3h it occurred at 69 degrees C. After irradiating for longer times (20-96h) the helix coil transition peak occurred at much lower temperatures. The peak was very broad and suggested that collagen was reduced by UV to different polypeptides of different molecular weight and different lower thermal stabilities. The irradiation of lens capsules with UVB light in vitro resulted in changes in the thermal properties of type-IV collagen consistent with increased cross-linking. DSC of lens capsules showed two major peaks at melting temperatures at 54 degrees C Tm1 and 78 degrees C Tm2, which can be attributed to the denaturation of the triple helix and 7S domains, respectively. UVB irradiation of lens capsules in vitro for 6 h caused an increase in Tm1 from 54 to 57 degrees C. The higher temperature required to denature the type-IV collagen after irradiation in vitro suggested an increase of intermolecular cross-linking. PMID- 16038808 TI - Fluorescence quenching and time-resolved fluorescence studies on Trichosanthes dioica seed lectin. AB - Fluorescence quenching and time-resolved fluorescence studies have been carried out on the Trichosanthes dioica seed lectin (TDSL). The emission lambdamax of native TDSL, seen at 328nm, shifts to 343nm upon denaturation with 6M guanidinium chloride. Quenching titrations were performed with neutral (acrylamide and succinimide) and ionic (I(-) and Cs(+)) quenchers in order to probe the exposure and accessibility of tryptophan residues of the protein. Maximum quenching was observed with acrylamide, followed by succinimide, iodide and Cs(+). Dramatic increase in the extent of quenching and other quenching parameters by all the quenchers were observed upon denaturation of TDSL, suggesting that all the tryptophan residues in native TDSL are buried in the hydrophobic core of the protein. Increase in the extent of quenching upon denaturation of TDSL was maximum with I(-) and minimum with Cs(+), suggesting the presence of positively charged residue(s), near at least one tryptophan residue. Addition of saccharide ligands such as methyl-beta-d-galactopyranoside and lactose led to a small, but reproducible decrease in the fluorescence intensity of the lectin. The presence of lactose provided a partial protection against quenching by I(-), Cs(+) and succinimide, but not acrylamide. In time-resolved fluorescence measurements the fluorescence decay curves could be best fitted to biexponential patterns with lifetimes of 4.09 and 1.53ns for native lectin, 3.40 and 1.65ns for the lectin in presence of 0.1M lactose and 3.50 and 1.40ns for denatured lectin. PMID- 16038809 TI - Molecular organization of the antifungal and anticancer drug 2-(2,4 dihydroxyphenylo)-5,6-dichlorobenzothiazole in solution and in monolayers: an effect of pH. AB - 2-(2,4-Dihydroxyphenylo)-5,6-dichlorobenzothiazol (dHBBT) is a new anticancer, antifungal and antibacterial drug characterised also by cytostatic and anticancer activity. The effect of pH on the molecular organization of dHBBT in monomolecular layers and in solution has been studied by electronic absorption and FT-IR spectroscopies. The analysis of the spectroscopic data suggests that at neutral and at high pH values (pH 6-8) dHBBT appears in the anionic form that prevents the formation of dimers due to the electrostatic repulsion between the molecules. PMID- 16038810 TI - Treatment of empyema. PMID- 16038811 TI - The mystique of organ transplantation. PMID- 16038812 TI - Assessment of patient functional status after surgery. AB - BACKGROUND: Improvement in day-to-day functioning is a valued outcome of surgical intervention. A new functional status assessment instrument, the Activities Assessment Scale (AAS), was designed for a randomized clinical trial evaluating laparoscopic versus open hernia repair procedures. STUDY DESIGN: The study data set included 2,164 patients at baseline and 1,562 patients at 3-month followup. Only male patients were enrolled in the trial. The psychometric characteristics of the AAS were examined in statistical analyses of cross-sectional and longitudinal data from the trial. Correlational analyses, factor analyses, and t tests were used to evaluate scale performance. RESULTS: We found that the AAS was a reliable measure (Cronbach's Coefficient Alpha =0.85) in the patient population studied. Factor analyses identified three subscales (sedentary activities; ambulatory activities; work and exercise activities). Construct validity was demonstrated by a correlation of 0.65 between the AAS and the physical functioning (PF) dimension of the SF-36 (p < 0.001); comparisons between clinical subgroups further confirmed its validity (p < 0.001). Patients reporting improvement on the physical functioning dimension after surgery showed an effect size of 1.20 for preoperative-postoperative change in their AAS scores. CONCLUSIONS: The AAS has been demonstrated to be a reliable, valid, and clinically responsive instrument that can be used to evaluate patient functioning after hernia surgery. It is easy to administer and requires less than 5 minutes of patient time to complete. This measurement system may prove useful in assessing surgical outcomes in both research and office practice settings. PMID- 16038813 TI - Failure of observation of blunt splenic injury in adults: variability in practice and adverse consequences. AB - BACKGROUND: The Eastern Association for the Surgery of Trauma Multiinstitutional Workgroup reported a failure rate for nonoperative management of blunt splenic injury in adults of 10.8%. Sixty percent of the failures occurred within 24 hours of admission. The purpose of this multiinstitutional study by the Eastern Association for the Surgery of Trauma was to determine common variables in failure of nonoperative management of blunt splenic injury in adults. STUDY DESIGN: Medical records were reviewed in a blinded fashion on 78 patients in whom nonoperative management failed. Statistical analysis was performed with ANOVA, extended chi-square, and Fisher's exact test; statistical significance was p<0.05. RESULTS: The 78 patients were categorized based on hemodynamic status. Forty-four percent were stable; 31% had transient hypotension or tachycardia that resolved with fluid infusion (responders); and 25% were unstable. Two-thirds of the unstable patients required laparotomy within 12 hours of admission; all had laparotomy within 72 hours. Mortality was significantly different when comparing the unstable to the stable and responder groups: stable (3%), responders (8%), and unstable (37%), despite similar age and only modest differences in Injury Severity Score. Eight CT scans were misinterpreted initially. Of 26 Focused Abdominal Sonography for Trauma (FAST) studies, 11 (42.3%) were false negative. Abnormal abdominal findings were noted in 67.7% of patients on admission. Ten patients died (12.8%). Sixty percent of the deaths were caused largely by delayed treatment of splenic or other abdominal injuries; one patient died in the responder group and five unstable patients died. CONCLUSIONS: Thirty percent to 40% of the patients who had unsuccessful nonoperative management in this study were selected inappropriately, with hemodynamic instability or initial misinterpretation of diagnostic studies. As a consequence, the majority of the deaths were from delayed treatment of intraabdominal injuries. This article suggests that written protocols, better adherence to sound clinical judgment, and experienced and timely interpretation of radiologic studies would reduce the incidence of failure of nonoperative management of blunt splenic injury in adults. PMID- 16038814 TI - A second malignancy is the major cause of death among thoracic squamous cell esophageal cancer patients negative for lymph node involvement. AB - BACKGROUND: The aim of the present study was to determine the major causes of death among esophageal cancer patients whose lymph nodes did not show metastasis at the time they received esophagectomy, and to consider strategies for improving survival rates among these patients. STUDY DESIGN: Between 1989 and 1999, 93 of our patients who underwent curative esophagectomy with extended lymph node dissection for thoracic squamous cell esophageal cancer showed no lymph node metastasis. We followed up these node-negative patients for as long as 10 years and determined the major causes of death. RESULTS: Sixty-three patients were still alive after esophagectomy, although 30 had died. Six patients died within 3 years after esophagectomy as a direct result of recurrence of their esophageal cancer; 13 died as a result of a second (extraesophageal) malignancy. Within the first 3 years, the major causes of death were recurrence (35%) and the second malignancy (35%); thereafter, the major cause was only the second malignancy (54%). There was no difference in the survival rates among patients with earlier, synchronous, or subsequent malignancies. Univariate and multivariate analyses of the 10-year survival showed the independent prognostic factors to be recurrence of esophageal cancer and development of a second malignancy, which respectively increased the risk of death 6.4 and 2.7 times. CONCLUSIONS: The major cause of reduced survival among thoracic squamous esophageal cancer patients, whose lymph nodes did not show metastasis, was a second malignancy. New strategies aimed at preventing or treating synchronous and subsequent malignancies could prolong the survival of these patients. PMID- 16038815 TI - Accuracy of intraoperative frozen-section analysis of breast cancer lumpectomy bed margins. AB - BACKGROUND: My colleagues and I have been using intraoperative frozen-section analysis (FSA) to evaluate lumpectomy margins in an attempt to reduce the number of additional operations that patients with ductal carcinoma in situ or stage I and II breast cancer would have to endure. We review our experience in breast conservation therapy (BCT) at the University of Florida (Gainesville) to determine the effectiveness of this approach. STUDY DESIGN: Operative reports, operative logs, and pathology reports were retrospectively reviewed for patients who had BCT from January 2001 to January 2004. Ninety-seven patients (116 operations) were reviewed. RESULTS: Nineteen patients required an additional operation (19.6%). Forty-three patients had positive margins on paraffin-embedded histologic analysis (44.3%). Accuracy of FSA was 84% when evaluated on a per-case basis, and 96% on a per-slide basis. False negatives were identified in 22 patients, affecting the operative pathway of 19 patients (19.6%) and were identified more frequently in cases of ductal carcinoma in situ (p < 0.001). There were no false positives. Additional operative time required for FSA was approximately 13 minutes per case. Eighty-four (86.6%) patients had successful BCT and 13 patients (13.4%) required mastectomy. CONCLUSIONS: Intraoperative analysis of margins using FSA is effective at minimizing the number of additional operations, with 19 patients benefiting from immediate intervention in this study. The authors believe that the number of second operations prevented and the high BCT rates justify performing FSA. Ductal carcinoma in situ is more difficult to identify in FSA. Preoperative discussions with the patient should reflect these findings. PMID- 16038817 TI - Surgical treatment of large contaminated abdominal wall defects. AB - BACKGROUND: Repair of a large, severely contaminated abdominal wall defect is a challenging problem. Most patients are currently treated with a multistaged procedure, which is time consuming, carries a high complication rate, and is often not finalized. STUDY DESIGN: In this study, our experience with a one-stage repair of contaminated abdominal wall defects using the Components Separation Method was evaluated with respect to morbidity and recurrence. Medical records of patients with contaminated abdominal wall defects, treated with the Components Separation Method from 1996 to 2000, were studied. Patients were invited to visit the outpatient clinic for a physical examination. RESULTS: Twenty-six patients with a median age of 49 years and a mean defect size of 267 cm2 were treated. Intraoperative contamination, graded according to the National Research Council (NRC), showed 22 National Research Council III patients and 4 National Research Council IV patients. Postoperatively, five superficial wound infections, three cases of pneumonia, three instances of recurrent enterocutaneous fistulation, and two cases of sepsis were observed. One of the patients with sepsis died after anastomotic disruption led to peritonitis and multiple organ failure. Two asymptomatic recurrences were diagnosed (8%) after a median followup of 27 months. CONCLUSIONS: Large contaminated abdominal wall hernias can be closed by the Components Separation Method, with a low recurrence rate but considerable morbidity. PMID- 16038816 TI - The training needs and priorities of male and female surgeons and their trainees. AB - BACKGROUND: Over the past decade, interest in general surgery careers has declined and the number of female medical school graduates has increased. This study was performed to identify the needs of both male and female surgical trainees and to guide design of training programs because attracting medical students to, and maintaining residents in, general surgery training programs can be difficult without a clear understanding of the training needs and priorities of both men and women. We hypothesized that men and women would express similar training priorities, yet have subjectively different experiences. STUDY DESIGN: Medical students, surgical residents, fellows, and fully trained surgeons affiliated with at least one of four major surgical societies were asked to complete a level-specific survey located on the American College of Surgeons Web site. RESULTS: There were 4,308 respondents (76% men). Men and women selected similar reasons for choosing a surgical career and residency program and criteria critical to a successful residency program, with women placing greater emphasis on clerkship experience and faculty diversity. There were no statistically significant differences between the men and women's perceptions of their own training. Although, when asked to evaluate whether certain aspects of training were comparable for male and female residents, women were statistically less likely to agree that their experiences were comparable with those of their male colleagues. CONCLUSIONS: Male and female surgical residents, fellows, and trained surgeons identified almost identical training needs and priorities yet women perceived disparate treatment. PMID- 16038818 TI - The contribution of laparoscopy in evaluation of penetrating abdominal wounds. AB - BACKGROUND: Penetrating abdominal wounds are traditionally explored by laparotomy. We investigated prospectively the role of laparoscopy within a defined protocol for management of penetrating abdominal wounds to determine its safety and advantages over traditional operative management. STUDY DESIGN: The study inclusion criteria were: stab and gun shot abdominal wounds, including junction zone injuries; stable vital signs; and absence of contraindications for laparoscopy. Diagnostic end points included detection of peritoneum or diaphragm violation, visceral injuries, and other indications for laparotomy. Systematic examination was undertaken using a multiport technique whenever the peritoneum or diaphragm had been violated. All repairs were done by open operation. RESULTS: A total of 40.6% of patients with penetrating trauma fulfilled study criteria (52 patients). Of these, 33% had no peritoneal penetration; 29% had no visceral injuries despite violation of peritoneum or diaphragm; 38% had visceral injuries, of which 40% (mainly liver and omentum) required no intervention. Twelve patients (23% of total) had open repairs. No missed injuries or death occurred in the study. Overall, 77% of penetrating injuries with stable vital signs avoided exploratory laparotomy. Compared with National Trauma Data Bank information for patients with the same Injury Severity Scores, hospitalization was reduced by more than 55% for the entire series. CONCLUSIONS: Laparoscopy for penetrating abdominal injuries in a defined set of conditions was safe and accurate, effectively eliminating nontherapeutic laparotomy and shortening hospitalization. PMID- 16038819 TI - Number versus distribution in classifying regional lymph node metastases from colon cancer. AB - BACKGROUND: Metastasis to regional lymph nodes from colon cancer is an important prognostic factor. In the TNM classification, node metastases are classified into three grades based on the number of metastatic nodes. In the Japanese General Rules for Clinical and Pathologic Studies on Cancer of the Colon, Rectum, and Anus (JGR), node metastases are classified into four grades based on the distribution of metastatic nodes. STUDY DESIGN: Based on the findings of node metastases in 164 patients with colon cancer obtained by the clearing method, node classifications by the JGR and TNM classifications were compared. RESULTS: The case distribution by the JGR grading was 41.5% in n (-), 29.3% in n1 (+), 18.3% in n2 (+), and 11.0% in n3 (+) disease. In the TNM classification, the distribution was 23.8% in pN1 and 34.8% in pN2 disease. The 5-year survival rate by the JGR was 98.4% in n (-), 74.3% in n1 (+), 51.2% in n2 (+), and 30.0% in n3 (+) disease; in TNM classification, this rate was 76.0% in pN1 and 45.0% in pN2 disease. CONCLUSIONS: In the classification of regional node metastases from colon cancer, the JGR classification showed a wider range in distribution and 5 year survival rate compared with the TNM system. PMID- 16038820 TI - Multicenter evaluation of the bovine mesenteric vein bioprostheses for hemodialysis access in patients with an earlier failed prosthetic graft. AB - BACKGROUND: The number of patients requiring hemodialysis increases each year, with a large cohort of patients still requiring prosthetic grafts for hemodialysis. All available prosthetic vascular access grafts have predictable failure rates, leading to a large group of patients with multiple failed access grafts. This report evaluates use of mesenteric vein bioprosthesis (MVB) as a conduit for patients who have failed at least one earlier synthetic vascular access graft. STUDY DESIGN: Two-hundred seventy-six access grafts were implanted in patients who had at least one earlier failed synthetic graft. Of these grafts, 183 were MVB and 93 were synthetic. Graft histories were obtained from 128 of the 183 patients who received the MVB, representing a nonrandomized historic data set of previously failed grafts as an internal control group (INT). Patency was determined by Kaplan-Meier analysis, and the Cox proportional hazards model was used for multivariate analysis of factors predictive of effect. RESULTS: Primary patency at 12 months was 35.6% MVB versus 28.4% synthetic grafts. At 24 months, secondary patency was 60.3% MVB, 42.9% synthetic, and 18.0% INT (p < 0.0001, log- rank). Complication rates, including dilation, seroma, infection, and thrombosis, were all notably lower for the MVB compared with synthetic grafts by Cox regression (p < 0.001). Intervention rate per patient year was lower in the MVB group (0.97 versus 1.37) compared with synthetic grafts (p = 0.003). CONCLUSIONS: MVB provided superior secondary graft patency compared with both historic graft data or newly implanted nonrandomized synthetic implants. A considerable reduction in thrombosis, infection, and interventions was observed with the MVB graft. These results suggest that MVB offers a safe alternative to patients who have a history of failing synthetic access grafts and may represent an option for extending vascular access to this patient population. PMID- 16038821 TI - Patterns of venous insufficiency after an acute deep vein thrombosis. AB - BACKGROUND: The purpose of this study was to investigate patterns of venous insufficiency during a 12-month period after an acute deep vein thrombosis. STUDY DESIGN: Seventy limbs in 67 patients with an acute deep vein thrombosis (DVT) involving 147 anatomic segments were evaluated with duplex scanning at 1 month, 3 months, 6 months, and 1 year. Venous segments were examined whether they were occluded, partially recanalized, or totally recanalized, and the development of venous reflux was evaluated. RESULTS: The segments investigated were the common femoral vein (38 segments), femoral vein (33 segments), popliteal vein (36 segments), and calf veins (40 segments). There were 35 limbs with isolated DVT and the remaining 35 had multisegment DVT. At 1 year, thrombi had fully resolved in 76% of the segments, 20% remained partially recanalized, and 5% were occluded. The venous occlusion was most predominant in the femoral vein (21%) at 1 year. On the contrary, rapid recanalization was obtained in calf veins than in proximal veins at each examination (p < 0.01). Deep vein insufficiency was detected as early as 1 month after development of DVT, and the reflux was most predominant in popliteal veins (56%), followed by femoral veins (18%). No reflux was found in calf veins. Multisegment DVTs had a significantly higher incidence of deep vein insufficiency than single segment DVTs at 1 year. Development of superficial venous insufficiency was found in 5 limbs (7%) and perforating vein insufficiency in 5 (7%). CONCLUSIONS: Lower extremity venous segments showed different proportions of occlusion, partial recanalization, and total recanalization. Calf veins showed more rapid recanalization than proximal veins. Venous reflux was noted as early as 1 month. The limbs involving multisegment DVTs on initial examination had a higher incidence of deep vein insufficiency and could require much longer followup studies. PMID- 16038822 TI - Pancreaticojejunal anastomosis is preferable to pancreaticogastrostomy after pancreaticoduodenectomy for longterm outcomes of pancreatic exocrine function. AB - BACKGROUND: The aim of this study was to evaluate pancreatic exocrine and endocrine function after pancreaticoduodenectomy. STUDY DESIGN: Pancreatic exocrine function was evaluated by a questionnaire and medical examination of stools after discontinuing pancreatic enzyme supplements for at least 10 days. Severe steatorrhea was defined as frequent, nauseating, yellow, and pasty stools, fecal output >200 g/d for more than 3 days. Endocrine function was evaluated by blood glucose level. Association between severe steatorrhea and age, indication, histologic obstructive pancreatitis, pancreaticojejunal anastomosis (PJA), pancreaticogastric anastomosis (PGA), and morbidity was studied. RESULTS: Fifty two patients underwent pancreaticoduodenectomy, complication rate was 33%. PJA was performed in 41 patients (79%) and PGA in 11 patients (21%). At a median followup of 75 months (24 to 156 months), 65% of the patients received pancreatic enzyme supplements. Severe steatorrhea was observed in 22 patients (42%). Incidence of postoperative diabetes was 14.6%. Patient age (more than 60 years), postoperative complication, and obstructive pancreatitis were not associated with postoperative severe steatorrhea. In cases of nonhistologic obstructive pancreatitis, PGA was more frequently associated with severe steatorrhea than PJA (70% versus 21.7%, p < 0.025). No factor significantly influenced incidence of postoperative diabetes. CONCLUSIONS: After pancreaticoduodenectomy, 42% of patients presented with severe steatorrhea. PJA allows better pancreatic exocrine function preservation than PGA and should be recommended. PMID- 16038823 TI - Visual analog scales for assessing surgical pain. AB - BACKGROUND: Four visual analog scales were constructed to assess sensory and affective components of operative pain. The Surgical Pain Scales (SPS) measure pain while at rest, pain during normal activities, pain during work or exercise, and pain unpleasantness. STUDY DESIGN: Longitudinal data from 2,164 patients in a randomized trial of laparoscopic versus open hernia repair established the reliability, validity, and sensitivity to change of the SPS. Correlations and t tests were used to determine their psychometric properties compared with the SF 36 health status instrument. RESULTS: Intraclass correlation coefficients of 0.95 for the sensory scales and 0.94 for the unpleasantness rating confirmed that the SPS produced reliable measurements. Correlations ranging from 0.44 to 0.60 between the visual analog scales and the bodily pain dimension on the SF-36 and significant differences between SPS levels for patients requiring more and less time to resume normal activities (p< or =0.015 to p< or =0.002) supported the validity of the scales. Clinical responsiveness was demonstrated by a 33.5-mm reduction (standard error = 1.4 mm) in the mean rating on a 150-mm scale measuring pain during normal activities for patients reporting postoperative improvement on the bodily pain dimension (p< or =0.0001). CONCLUSIONS: The Surgical Pain Scales demonstrated excellent psychometric properties in this study population. The SPS can be used to compare pain levels between groups at a single point in time or to track change for individual patients over time or after operations. Individualized pain management interventions can be tailored based on the sensory and effective ratings. PMID- 16038824 TI - Mortality and morbidity after resection for adenocarcinoma of the gastroesophageal junction: predictive factors. AB - BACKGROUND: Resection for adenocarcinoma of the gastroesophageal junction (AGEJ) is associated with severe mortality and morbidity. This retrospective study aimed to evaluate mortality and morbidity after resection for AGEJ and to determine their predictive factors. STUDY DESIGN: Data from 1,192 patients (mean age 65 +/- 11 years) who underwent resection for AGEJ by members of French Association of Surgery from 1985 to 2000 were collected. A stepwise logistic regression model was built to identify by multivariate analysis the variables independently associated with mortality, morbidity, anastomotic leakage, and major pulmonary complications. RESULTS: Distribution of Siewert's type was: I = 480 (40%), II = 500 (42%), and III = 212 (18%). Most type I and II tumors were treated by esophagectomy and proximal gastrectomy (93% and 58%, respectively), using an approach including a thoracotomy (82% and 64%, respectively); type III tumors were treated mainly by total gastrectomy and distal esophagectomy (83%), through an exclusive transabdominal approach (69%). Seventy-six (6%) patients died postoperatively. Only American Society of Anesthesiologists (ASA) scores III and IV (p < 0.001) and period of study (p = 0.025) were predictive of mortality. Predictive factors of overall morbidity (overall rate = 35%) were high ASA score (p < 0.001), age more than 60 years (p = 0.020), male gender (p = 0.039), and cervical anastomosis (p = 0.001). Factors predictive of anastomotic leakage (overall rate = 9%) were high ASA score (p = 0.006) and manual anastomosis (p = 0.010). Factors predictive of major pulmonary complications (overall rate = 23%) were high ASA score (p = 0.015), age more than 60 years (p < 0.001), anastomotic leakage (p < 0.001), and abdominal complications (p = 0.003). CONCLUSIONS: ASA score is a reliable predictive factor of operative mortality and morbidity after resection of AGEJ. PMID- 16038825 TI - A proposal for evaluation of platelet aggregation and adhesiveness for clinical trials with cyclooxygenase inhibitors and nonsteroidal antiinflammatory drugs. PMID- 16038826 TI - What's new in gynecology and obstetrics. PMID- 16038827 TI - Comprehensive endovascular therapy for femoropopliteal arterial atherosclerotic occlusive disease. PMID- 16038828 TI - Colon sentinel lymph node mapping: practical surgical applications. PMID- 16038829 TI - Obstructive uropathy from giant inguinal bladder and ureteral herniation. PMID- 16038830 TI - Destructive maxillomandibular brown tumor in severe hyperparathyroidism. PMID- 16038831 TI - Novel parastomal hernia repair using a modified polypropylene and PTFE mesh. PMID- 16038832 TI - Cervical spine fracture in the ankylosing spondylitis patient. PMID- 16038833 TI - Index of suspicion for pneumatocele formation and awareness of management is not just for the trauma surgeon. PMID- 16038834 TI - Posttraumatic pneumatocele. PMID- 16038836 TI - Sentinel lymph node (SLN) biopsy in clinically node positive disease. PMID- 16038838 TI - Contribution of autoantibody assays to the diagnosis of adulthood celiac disease. PMID- 16038839 TI - Spa therapy for ankylosing spondylitis: still useful? PMID- 16038840 TI - Toxic effects of nonsteroidal antiinflammatory drugs on the small bowel, colon, and rectum. AB - The gastrointestinal toxicity of conventional nonsteroidal antiinflammatory drugs (NSAIDs) is not confined to the stomach and proximal duodenum but extends also to the rest of the small bowel, colon, and rectum. Long-term NSAID therapy usually induces clinically silent enteropathy characterized by increased intestinal permeability and inflammation. Chronic occult bleeding and protein loss may result in iron-deficiency anemia and hypoalbuminemia. NSAIDs can also induce small bowel ulcers that infrequently lead to acute bleeding, perforation, or chronic scarring responsible for diaphragm-like strictures. At the colon and rectum, NSAID use can result in de novo lesions such as nonspecific colitis and rectitis, ulcers, and diaphragm-like strictures. NSAIDs have been implicated in the development of segmental ischemic colitis. In patients with diverticular disease, NSAID use increases the risk of severe diverticular infection and perforation. NSAIDs can trigger exacerbations of ulcerative colitis or Crohn's disease. With selective COX-2 inhibitors, the risk of gastrointestinal toxicity is reduced as compared to conventional NSAIDs but is not completely eliminated. Experimental studies suggest that long-term COX-2 inhibitor therapy may cause damage to the previously healthy small bowel. Similar to conventional NSAIDs, COX 2 inhibitors may be capable of triggering exacerbations of inflammatory bowel disease. PMID- 16038841 TI - Improvement of the clinical outcome in Ankylosing spondylitis by balneotherapy. AB - AIMS: This study is designed to show the efficacy of balneotherapy and balneotherapy (BT) + nonsteroid antiinflammatory drug (NSAID) use in Ankylosing spondylitis (AS) patients. METHODS: In this prospective study, BT, BT+ NSAID and NSAID therapy in 61 patients with AS were evaluated by ASAS core set. BT group (21 patients) was treated only with BT for 20 min, once a day, 5 days a week, over a period of 3 weeks. BT+NSAID group (20 patients) was treated with 1000 mg naproxen as well as BT. NSAID group (20 patients) was treated with 1000 mg naproxen. All of the participants did respiratory and postural exercises for 20 min a day and for the whole study period. Each patient was evaluated on admission (before treatment), at the end of the therapy and 6 months after the treatment. RESULTS: At the end of the study, statistically significant improvement was observed in all the clinical parameters of the patients in BT (G1), BT+NSAID (G2) and NSAID (G3) groups. This significant symptomatic and clinical improvement was maintained even 6 months after the treatment. The changes from baseline to follow up were similar in G1 and G2 except duration of morning stiffness (DMS) and chest expansion (CE). Improvements in CE and DMS were better in G1 and G2, respectively. Improvements observed in G1 and G2 were superior to the improvements observed in G3 for the variables of morning pain, nocturnal pain, DMS, global well being of the patient, occiput-wall distance, CE, finger to floor distance and functional index. In Schober test, improvement observed in G1 was statistically superior to G3. CONCLUSION: We concluded that BT can be suggested as an effective symptomatic treatment modality in patients with AS. Furthermore, sufficient improvement in clinical parameters can be obtained by BT alone. PMID- 16038842 TI - Infliximab continuation rates in patients with rheumatoid arthritis in everyday practice. AB - Infliximab is a major breakthrough in the management of rheumatoid arthritis (RA). We evaluated infliximab continuation rates and reasons for discontinuation in patients with RA. PATIENTS AND METHODS: We studied patients with RA started on infliximab at any time between March 2000 and December 2002, under the conditions of everyday practice (as opposed to clinical trial settings), as recommended by the French marketing license (3 mg/kg as an intravenous infusion at weeks 0, 2, and 6 then at 8-week intervals). The number of infliximab infusions, side effects, and nonresponse rates was recorded. The Kaplan-Meier method was used to evaluate treatment continuation. Reasons for discontinuation were studied. RESULTS: We included 41 patients, with a mean age of 54 years, a mean RA duration of 9 years, and a mean of three previous disease-modifying antirheumatic drug treatments. The total number of infliximab infusions was 461 with a mean of 10.8 per patient and a mean follow-up under treatment of 15.3 months. The proportions of patients still on infliximab were 82%, 74%, and 67% after 6, 12, and 24 months, respectively. The main reasons for discontinuation were escape phenomenon (n = 6, 42.8% of discontinuations) and allergy (n = 3); in one case each, the reason was primary ineffectiveness, severe infection, plans to start a pregnancy, poor compliance, and unavailability for follow-up. There were 59 recorded episodes of side effects, with a profile similar to that in the literature and in postmarketing databases. CONCLUSION: The infliximab continuation rate in everyday practice in patients with RA (67% after 2 years) was consistent with published data and with the results of controlled trials. PMID- 16038843 TI - Benefits of pamidronate in children with osteogenesis imperfecta: an open prospective study. AB - OBJECTIVES: To study the efficacy of pamidronate in children with osteogenesis imperfecta (OI). PATIENTS AND METHODS: Twenty-nine patients (median age 8.7 years), were given pamidronate in cyclic infusions of 3 days. Patients received 3 13 cycles (median 6), at a dose of 0.5 mg/kg/day in infants (below 2 years of age) and 1 mg/kg/day in children (2 years and older). The interval time between cycles was 2 months in infants and 4 months in children. The median follow-up was 16 months. All patients received daily supplementation of calcium, vitamin D and physical rehabilitation. Assessments were performed at baseline and before each cycle. Fracture rate under treatment was compared to the one in the pre-treatment period. RESULTS: Pain decreased after the first infusion cycle (P < 0.0001). The median of fracture incidence decreased from 15 to 0.5 per year in infants and from 2.0 to 1 per year in children (P = 0.04). Alkaline phosphatase decreased by 31.2% and N-telopeptide collagen cross-links decreased by 61.8% (P < 0.001). Bone mineral density (BMD) of the spine increased by a median of 55.4% (P < 0.001). Z scores increased from a median of -4.7 to -2.6 (P < 0.001). The femoral neck, BMD increased by a median of 16%. The area of the first four lumbar vertebrae increased by a median of 21.5% (P < 0.001). No adverse effect on growth or on fracture healing was observed. Side effects were symptomatic hypocalcemia in one infant, and the transient acute phase reaction. CONCLUSION: Pamidronate increases BMD, decreases bone remodeling markers, pain and fracture rate in infants and children with OI. PMID- 16038844 TI - Chronic osteomyelitis of the metacarpals. Report of a case. AB - Chronic osteomyelitis of the hand is uncommon and affects the metacarpals in only 3% of cases. We report a case of chronic osteomyelitis involving two metacarpals, and we present a review of the relevant literature. CASE-REPORT: A 41-year-old man with a 5-year history of psoriatic arthritis was admitted for a swelling over the dorsum of the left hand. At admission, he was in good general health and had no fever. In addition to the swelling, he had synovitis of the right ankle and psoriasis over the hands and elbows. The spine and sacroiliac joints were normal to physical examination. The erythrocyte sedimentation rate was 110 mm/h, the C reactive protein level was 48 mg/l, and the leukocyte count was 9600/mm3. A radiograph of the hands disclosed a bone-within-bone image in the second and third metacarpals of the left hand and arthritis of the left carpal joints. A fluid collection over the dorsum of the left hand was visualized by ultrasonography. Aspiration recovered serous fluid that contained no organisms by microscopic examination or culture. Investigations for tuberculosis and a serological test for HIV infection were negative. Computed tomography showed a florid periosteal reaction encasing the diaphyses of the second and third metacarpals and enclosing bony sequestra; abnormal carpal bone architecture and thickening of the soft tissues related to joint effusions were seen also. The diagnosis was chronic osteomyelitis of the metacarpals. Two antimicrobials active against staphylococci were given and the bony sequestra were removed surgically. Histological examination of the operative specimens showed nonspecific osteitis. After 3 months of treatment, the outcome was favorable. CONCLUSION: Chronic osteomyelitis of the metacarpals is exceedingly rare but results in severe functional incapacitation and major social and economic burdens. Our case illustrates an unusual pattern with involvement of two metacarpals in the same hand. An early diagnosis followed by prompt treatment increases the likelihood of a favorable outcome. PMID- 16038845 TI - Acitretin-induced enthesitis in a patient with psoriatic arthritis. AB - A patient with psoriatic arthritis and cutaneous psoriasis took acitretin for 10 years to treat his skin lesions. Radiographs disclosed exuberant ossifications in several entheses. Their features were not typical for psoriatic arthritis but were consistent with acitretin-induced hyperostosis. Retinoids are known to induce hyperostosis, most notably when they are used in high dosages and over long periods. The concomitant presence of two conditions affecting the entheses may explain the exuberant nature of the ossifications in our patient. PMID- 16038846 TI - Severe pyogenic infections in patients taking infliximab: a regional cohort study. AB - OBJECTIVE: To evaluate the prevalence and risk factors of severe pyogenic infections in rheumatology patients taking infliximab in everyday practice. METHODS: Regional prospective cohort study of patients taking infliximab for rheumatoid arthritis or ankylosing spondylitis with data collection on standardized forms. The medical records of patients with severe pyogenic infections were subjected to a detailed retrospective review. Patients with and without severe pyogenic infections were compared. RESULTS: The cohort included 83 patients (55 women and 28 men). Severe pyogenic infections occurred in five (6%) patients (three women and two men), all of whom had acute or underlying risk factors. Higher values were found in these five patients for mean age (65.8 +/- 12 vs. 53.9 +/- 13 years, P = 0.04) and mean daily glucocorticoid dosage (15.5 +/ 9 vs. 6.9 +/- 7 mg/day prednisone-equivalent, P = 0.036), as compared to the other patients. CONCLUSION: Older age and high-dose glucocorticoid therapy are associated with an increased risk of severe pyogenic infection during infliximab therapy. Caution is in order when starting and monitoring infliximab therapy in patients with risk factors. Our data also emphasize the need for a careful search for risk factors before each infliximab infusion. PMID- 16038847 TI - A rare cause of painful cervical swelling: myositis ossificans progressiva in childhood. Report of a case. AB - Myositis ossificans progressiva (MOP) is a rare condition of which we report a case in a 13-year-old girl with involvement of the cervical paraspinal and periscapular muscles. No ectopic ossifications were visible on plain radiographs. Computed tomography disclosed diffuse inflammation. Plain radiographs of the toes confirmed the diagnosis by visualizing characteristic bone abnormalities. Knowledge of the digital abnormalities seen in MOP is important to avoid unnecessary diagnostic investigations, most notably a surgical biopsy, which may trigger a flare of the disease. The prognosis is guarded. PMID- 16038848 TI - Evaluation of the fracture threshold by quantitative ultrasound densitometry in everyday practice. PMID- 16038849 TI - Scleroderma complicated with tamponade during pregnancy. PMID- 16038850 TI - Fracture healing in a patient with osteopoikilosis. PMID- 16038853 TI - Improving application of neurodynamic (neural tension) testing and treatments: a message to researchers and clinicians. PMID- 16038851 TI - Comments about the editorial by Benedicte Mugnier and Jean Roudier entitled "Factors predicting responsiveness to anti-TNFalpha therapy in patients with rheumatoid arthritis: using biotherapies rationally". PMID- 16038854 TI - Reliability of palpation of humeral head position in asymptomatic shoulders. AB - The purpose of this study was to determine if, within a normal population: (1) palpation of the humeral head, relative to the acromion, in three static positions, was a reliable technique (2) there was a difference in humeral head position between the dominant and non-dominant shoulders in the three positions (3) there was a difference in humeral head position relative to the acromion between the arm at side (AS), the 90 degrees abduction/external rotation (AER) and 90 degrees abduction/internal rotation (AIR) positions. This test-retest study recorded palpation landmarks using a standardized protocol. Intra-tester reliability was above 0.8 for both AS and AER and all other ICCs were below 0.6. There was no systematic difference between dominant and non-dominant sides in any of the three positions (AS P=0.408, AER P=0.448, AIR P=0.233). There was a significant difference in measurements between each position (P<0.001). It can be concluded that, palpation of humeral head position in relation to the acromion is a reliable technique in the AS position. These normative data provide a baseline that can be used for future comparison if differences are found to exist in subgroups with pathological shoulder conditions where larger glenohumeral translations are thought to exist. PMID- 16038855 TI - A normative database of lumbar spine ranges of motion. AB - The overall aim of the work was to develop a comprehensive normative database of indices for ranges of motion in the lumbar spine, in an asymptomatic sample of the general population. This was a repeated measures prospective study utilizing a reliable and valid instrument, the modified CA6000 Spine Motion Analyzer (Orthopedic Systems Inc. Union City CA & Troke/University of Brighton). The portable equipment was used to collect data in a variety of community settings (e.g. schools, GP surgeries, offices, leisure centres, emergency services stations). A total of 405 asymptomatic subjects (196 female, 209 male) aged 16-90 yr from sedentary, mixed and physically demanding occupations participated in the study and data were collected in standing, at different times of the day, following a standardized methodology for lumbar spine motion in the sagittal, coronal and horizontal planes. Age-related centile graphs were derived separately for male and female subjects in flexion, extension, left and right lateral flexion and left and right axial rotation. All 12 graphs are presented as an appendix located on the Manual Therapy website. Overall, flexion (73-40 degrees) and lateral flexion (28-14 degrees, L&R) declined 45% and 48%, respectively, across the age range. Extension (29-6 degrees) declined the greatest at 79%. By contrast, no overall decline in axial rotational RoMs was recorded, and the median RoM remained at 7 degrees each way across the age spectrum examined. A comprehensive database of indices of lumbar spine ranges of motion has thus been developed which is gender specific, age related, drawn from a wide age range and presents data for all three planes of motion. It is considered that the new database has a number of potential clinical and research applications. PMID- 16038856 TI - Diagnosis of sacroiliac joint pain: validity of individual provocation tests and composites of tests. AB - Previous research indicates that physical examination cannot diagnose sacroiliac joint (SIJ) pathology. Earlier studies have not reported sensitivities and specificities of composites of provocation tests known to have acceptable inter examiner reliability. This study examined the diagnostic power of pain provocation SIJ tests singly and in various combinations, in relation to an accepted criterion standard. In a blinded criterion-related validity design, 48 patients were examined by physiotherapists using pain provocation SIJ tests and received an injection of local anaesthetic into the SIJ. The tests were evaluated singly and in various combinations (composites) for diagnostic power. All patients with a positive response to diagnostic injection reported pain with at least one SIJ test. Sensitivity and specificity for three or more of six positive SIJ tests were 94% and 78%, respectively. Receiver operator characteristic curves and areas under the curve were constructed for various composites. The greatest area under the curve for any two of the best four tests was 0.842. In conclusion, composites of provocation SIJ tests are of value in clinical diagnosis of symptomatic SIJ. Three or more out of six tests or any two of four selected tests have the best predictive power in relation to results of intra-articular anaesthetic block injections. When all six provocation tests do not provoke familiar pain, the SIJ can be ruled out as a source of current LBP. PMID- 16038857 TI - Intra- and inter-rater reliability of the anterior atlantodental interval measurement from conventional lateral view flexion/extension radiographs. AB - An investigation of intra- and inter-rater reliability anterior atlantodental interval (AADI) measurements was conducted using flexion/extension plain radiographs. Flexion and extension lateral radiographs of individuals investigated for atlantoaxial instability were measured for AADI on three occasions. Intra-rater intraclass correlation coefficients (ICC) were calculated for both flexion (0.99) and extension (0.96). Inter-rater ICCs were 0.93 and 0.84 for flexion and extension, respectively. The AADI measurement proved to be reproducible with a minimal standard of error, between and within raters. PMID- 16038858 TI - The use of manipulation in a patient with an ankle sprain injury not responding to conventional management: a case report. PMID- 16038859 TI - Clinical anatomy serving manual therapy, by Mercer SR, Rivett DA Manual Therapy 9 (2) (2004) 59. PMID- 16038860 TI - The dog and its tail. PMID- 16038861 TI - Commencement, challenge, and uncertainty--responding to the future of health care. PMID- 16038862 TI - Pfeiffer syndrome: systemic and ocular implications. AB - BACKGROUND: In 1964, Pfeiffer described a three-generation family in which eight individuals had a syndrome consisting of craniosynostosis, broad thumbs and great toes, and partial syndactyly of the hands and feet. Pfeiffer syndrome affects males and females equally, and is most commonly a result of de novo mutations, but can be inherited in an autosomal dominant fashion. Pfeiffer syndrome is considered Type V of the five acrocephalosyndactly syndromes (ACS), a group of rare genetic diseases that involve premature closure of the cranial sutures. Cohen, in 1993, further described Pfeiffer syndrome and it's various expression patterns by creating three subgroups of the syndrome. CONCLUSIONS: While Pfeiffer syndrome is clearly a rare disorder, affecting 15 of every 1 million births, there has been a series of publications reviewing the difficult differential diagnosis among Pfeiffer types and between the other acrocephalosyndactly syndromes. While these publications individually focus on a variety of specific systemic and ocular implications of the syndrome, together they encompass the scope of the syndrome. Since Pfeiffer syndrome mainly affects the craniofacial regions, the eye care professional plays an essential role in diagnosis and management. What follows are guidelines to aid in the diagnosis, ophthalmic and functional testing, and management of this disorder. PMID- 16038863 TI - Non-ptotic ocular myasthenia gravis: a common presentation of an uncommon disease. AB - BACKGROUND: Myasthenia gravis (MG) is an acquired autoimmune disease of the neuromuscular junction which causes rapid muscle fatigue and weakness. Two thirds of all cases of myasthenia gravis (MG) initially manifest ptosis. In the absence of the characteristic variable ptosis, MG can present a challenge to the clinician. This article will review the current diagnostic and management strategies for MG. CASE REPORTS: Five cases will be presented that did not initially present with ptosis. Each of these cases was previously misdiagnosed as a result of presentation of atypical myasthenia gravis signs and symptoms. The first two cases had signs and symptoms of a typical accommodative/vergence anomaly. The others manifested diplopia not normally associated with MG: one had a noncomitant vertical deviation; another had a stable 6(th) nerve palsy; and the third had a basic esotropia. CONCLUSION: Although the hallmark findings of MG are ptosis and eye muscle palsy with variability, MG may present without ptosis, affect nonstriated muscles, and/or manifest either as a nonstrabismic vergence anomaly or as comitant nonvariable strabismic deviation. PMID- 16038864 TI - Evaluation and management of plateau iris syndrome: case report and review. AB - BACKGROUND: This article documents a case of plateau iris syndrome and provides a review of the current concepts regarding the pathogenesis, diagnosis, and management of this condition. CASE REPORT: A patient who had patent peripheral iridotomies O.D. and O.S. to treat previous episodes of acute angle closure came to clinic with unilaterally elevated intraocular pressures, narrow anterior chamber angles, and a flat iris configuration. The patient was diagnosed with plateau iris syndrome. CONCLUSIONS: The clinician must be aware of, and be able to distinguish, the clinical signs that are characteristic of plateau iris syndrome. An understanding of the mechanisms involved in the development of this type of primary angle closure may facilitate its appropriate diagnosis and management. PMID- 16038865 TI - Automobile air bags: friend or foe? A case of air bag-associated ocular trauma and a related literature review. AB - BACKGROUND: Although air bags are placed in automobiles to act as safety devices, they have been shown to carry a risk of injury themselves. Ocular injury, in particular, can often be a direct consequence of air bag deployment. A case of ocular air bag injury is presented. A discussion and review of the current literature on this issue follows. CASE REPORT: A 63-year-old man was transferred to our clinic after sustaining injuries related to a motor vehicle accident, during which the automobile's air bag was deployed. Initial examination revealed many signs of blunt ocular trauma of the O.D., including iridodialysis, dislocated lens with traumatic cataract, and traumatic/inflammatory glaucoma. Initial B-scan showed an attached retina O.D. One month later, the patient underwent an attempted pars plana vitrectomy with lensectomy, iris repair, and insertion of an anterior chamber intraocular lens. Complications arose during the procedure, and a total retinal detachment developed. Resultant acuity is no light perception O.D. CONCLUSIONS: Although ocular morbidity can be a direct consequence of air bag deployment, most eye injuries are minimal, and seem to be outweighed by the benefits of air bags. Drivers, as well as passengers, can minimize associated injuries by adhering to specific safety guidelines. This, as well as continual modification and improvement in air bag design, will maximize the safety of air bags and decrease the incidence of vision-threatening ocular injury caused by air bag deployment. PMID- 16038866 TI - Developmental Eye Movement Test: reliability and symptomatology. AB - BACKGROUND: The Developmental Eye Movement Test (DEM) is a widely used visual skill test, especially in the context of a vision therapy evaluation. It is intended to diagnose oculomotor dysfunction (OMD) and can also identify deficient rapid automatized naming. As such, its reliability and associated symptomatology are important. METHODS: The DEM test-retest reliability was investigated within two populations: a group of 53 office patients who were participating in vision therapy evaluation in a private optometry practice, and a smaller group of 13 subjects at their school. One to four weeks separated the test and retest for both groups. We also studied the relationship between results on a questionnaire of symptoms associated with OMD and DEM test performance in these two populations. RESULTS: The first administration of the DEM significantly correlated with the second for all four of its scores for both groups. The office group had higher intra-class correlation coefficients than the school group. There was good agreement between test and re-test in terms of pass-fail classification for the office group. Symptomatic subjects performed poorer than asymptomatic subjects on the DEM. Failing the DEM Ratio, the score used to diagnose OMD, identified 90% of the subjects who were symptomatic. CONCLUSIONS: The DEM has good intra-subject test-retest reliability for all four of its scores when it is administered in an office setting to patients participating in a vision therapy evaluation. It also has good consistency in classifying patients as pass or fail. Performance on the DEM relates to certain symptoms that are associated with OMD. PMID- 16038867 TI - Influence of cell-model boundary conditions on the conductivity and electrophoretic mobility of concentrated suspensions. AB - In the last few years, different theoretical models and analytical approximations have been developed addressing the problem of the electrical conductivity of a concentrated colloidal suspension. Most of them are based on the cell model concept, and coincide in using Kuwabara's hydrodynamic boundary conditions, but there are different possible approaches to the electrostatic boundary conditions. We will call them Levine-Neale's (LN, they are Neumann type, that is they specify the gradient of the electrical potential at the boundary), and Shilov-Zharkikh's (SZ, Dirichlet type). The important point in our paper is that we show by direct numerical calculation that both approaches lead to identical evaluations of the conductivity of the suspensions if each of them is associated to its corresponding evaluation of the macroscopic electric field. The same agreement between the two calculations is reached for the case of electrophoretic mobility. Interestingly, there is no way to reach such identity if two possible choices are considered for the boundary conditions imposed to the field-induced perturbations in ionic concentrations on the cell boundary (r = b), deltan(i) (r = b). It is demonstrated that the conditions deltan(i)(b) = 0 lead to consistently larger conductivities and mobilities. A qualitative explanation is offered to this fact, based on the plausibility of counter-ion diffusion fluxes favoring both the electrical conduction and the motion of the particles. PMID- 16038868 TI - Differential responses of PPARalpha, PPARdelta, and PPARgamma reporter cell lines to selective PPAR synthetic ligands. AB - To characterize the specificity of synthetic compounds for peroxisome proliferator-activated receptors (PPARs), three stable cell lines expressing the ligand binding domain (LBD) of human PPARalpha, PPARdelta, or PPARgamma fused to the yeast GAL4 DNA binding domain (DBD) were developed. These reporter cell lines were generated by a two-step transfection procedure. First, a stable cell line, HG5LN, expressing the reporter gene was developed. These cells were then transfected with the different receptor genes. With the help of the three PPAR reporter cell lines, we assessed the selectivity and activity of PPAR agonists GW7647, WY-14-643, L-165041, GW501516, BRL49653, ciglitazone, and pioglitazone. GW7647, L-165041, and BRL49653 were the most potent and selective agonists for hPPARalpha, hPPARdelta, and hPPARgamma, respectively. Two PPAR antagonists, GW9662 and BADGE, were also tested. GW9662 was a selective PPARgamma antagonist, whereas BADGE was a low-affinity PPAR ligand. Furthermore, GW9662 was a full antagonist on PPARgamma and PPARdelta, whereas it showed partial agonism on PPARalpha. We conclude that our stable models allow specific and sensitive measurement of PPAR ligand activities and are a high-throughput, cell-based screening tool for identifying and characterizing PPAR ligands. PMID- 16038869 TI - cRNA target preparation for microarrays: comparison of gene expression profiles generated with different amplification procedures. AB - Microarray technology has become a standard tool for generation of gene expression profiles to explore human disease processes. Being able to start from minute amounts of RNA extends the fields of application to core needle biopsies, laser capture microdissected cells, and flow-sorted cells. Several RNA amplification methods have been developed, but no extensive comparability and concordance studies of gene expression profiles are available. Different amplification methods may produce differences in gene expression patterns. Therefore, we compared profiles processed by a standard microarray protocol with three different types of RNA amplification: (i) two rounds of linear target amplification, (ii) random amplification, and (iii) amplification based on a template switching mechanism. The latter two methods accomplish target amplification in a nonlinear way using PCR technology. Starting from as little as 50 ng of total RNA, the yield of labeled cRNA was sufficient for hybridization to Affymetrix HG-U133A GeneChip array using the respective methods. Replicate experiments were highly reproducible for each method. In comparison with the standard protocol, all three approaches are less sensitive and introduced a minor but clearly detectable bias of the detection call. In conclusion, the three amplification protocols used are applicable for GeneChip analysis of small tissue samples. PMID- 16038870 TI - An immunosorbent, nonradioactive p38 MAP kinase assay comparable to standard radioactive liquid-phase assays. PMID- 16038871 TI - Modulation of photosynthetic electron transport in the absence of terminal electron acceptors: characterization of the rbcL deletion mutant of tobacco. AB - Tobacco rbcL deletion mutant, which lacks the key enzyme Rubisco for photosynthetic carbon assimilation, was characterized with respect to thylakoid functional properties and protein composition. The Delta rbcL plants showed an enhanced capacity for dissipation of light energy by non-photochemical quenching which was accompanied by low photochemical quenching and low overall photosynthetic electron transport rate. Flash-induced fluorescence relaxation and thermoluminescence measurements revealed a slow electron transfer and decreased redox gap between Q(A) and Q(B), whereas the donor side function of the Photosystem II (PSII) complex was not affected. The 77 K fluorescence emission spectrum of Delta rbcL plant thylakoids implied a presence of free light harvesting complexes. Mutant plants also had a low amount of photooxidisible P700 and an increased ratio of PSII to Photosystem I (PSI). On the other hand, an elevated level of plastid terminal oxidase and the lack of F0 'dark rise' in fluorescence measurements suggest an enhanced plastid terminal oxidase-mediated electron flow to O2 in Delta rbcL thylakoids. Modified electron transfer routes together with flexible dissipation of excitation energy through PSII probably have a crucial role in protection of PSI from irreversible protein damage in the Delta rbcL mutant under growth conditions. This protective capacity was rapidly exceeded in Delta rbcL mutant when the light level was elevated resulting in severe degradation of PSI complexes. PMID- 16038872 TI - Structure-affinity relationship in the interactions of human organic anion transporter 1 with caffeine, theophylline, theobromine and their metabolites. AB - It is well known that human organic anion transporter 1 (hOAT1) transports many kinds of drugs, endogenous compounds, and toxins. However, little is known about the structure-affinity relationship. The aim of this study was to elucidate the structure-affinity relationship using a series of structurally related compounds that interact with hOAT1. Inhibitory effects of xanthine- and uric acid-related compounds on the transport of p-aminohippuric acid were examined using CHO-K1 cells stably expressing hOAT1. The order of potency for the inhibitory effects of xanthine-related compounds on PAH uptake was 1-methyl derivative>7-methyl derivative>3-methyl derivative falling dotsxanthine>1,3,7-trimethyl derivative (caffeine). The order of potency of the inhibition was 1,3,7-trimethyluric acid>1,3-dimethyluric acid>1,7-dimethyluric acid>1-methyluric acid>uric acid. A significant correlation between inhibitory potency and lipophilicity of the tested uric acid-related compounds was observed. The main determinant of the affinity of xanthine-related compounds is the position of the methyl group. On the other hand, lipophilicity is the main determinant of the affinity of uric acid-related compounds. PMID- 16038873 TI - Externalizing and personalizing biases in persecutory delusions: the relationship with poor insight and theory-of-mind. AB - The presence of externalizing bias (EB) for negative events together with personalizing bias (PB) (a bias to blame others rather than circumstances) may jointly constitute a vulnerability to develop persecutory delusions (PDs). Whereas EB purportedly serves to defend a vulnerable self-concept by avoiding negative self-attributions and might therefore exacerbate poor insight, PB may reflect cognitive deficits, including theory-of-mind impairment. We investigated these proposals in 34 schizophrenic patients with a history of PDs and 21 healthy controls. Patients with moderate- to severe-PDs and patients without a current PD showed excessive EB which was, surprisingly, absent in patients with mild persecutory delusions (mild-PDs). That EB might wax and wane with fluctuating delusional intensity was interpreted in accord with a new dynamic model of attribution self-representation cycles. As predicted, EB exacerbated poor insight. However, counter to predictions, theory-of-mind impairment did not increase PB, which was marked in all participants, whether clinical or non clinical; instead, theory-of-mind impairment was also correlated with poor insight. Our findings indicate multiple pathways to poor insight, one of which is a theory-of-mind difficulty, impairing the capacity to simulate other perspectives for the purpose of critically evaluating one's own beliefs and circumstances. PMID- 16038874 TI - One-year follow-up of pharmacotherapy-resistant patients with panic disorder treated with cognitive-behavior therapy: Outcome and predictors of remission. AB - Non-response to pharmacotherapy for panic disorder (PD) is a well-documented problem. However, little information exists to guide next-step strategies for these non-responders. In addition to pharmacologic augmentation strategies, several studies support the efficacy of cognitive-behavior therapy (CBT) for these patients, although data on long-term outcomes has been lacking. In this study, we provide one-year outcomes on a sample of 63 patients who completed group CBT for PD after failing to respond adequately to previous pharmacotherapy. Sustained significant benefit was found for all dimensional outcome scores, and nearly two-thirds of the sample met remission criteria. This occurred with reductions in medication use over the follow-up period. Negative predictors of remission status included comorbid dysthymia, social phobia, and generalized anxiety disorder. These results provide additional evidence for the efficacy of CBT for medication non-responders with PD. PMID- 16038875 TI - Expression and regulation of WISP2 in rheumatoid arthritic synovium. AB - Numbers of growth factors expressed in the synovium deeply impact on the pathology of rheumatoid arthritis (RA). The WISP family was identified as growth factors, which are upregulated by WNT signaling. In the present study, we investigated expression pattern and regulatory mechanisms of WISPs in the synovium in patients with RA and osteoarthritis (OA). Among three members of WISP family, WISP2 mRNA was only preferentially detected in RA synovium by RT-PCR. WISP2 expression was immunohistochemically identified in RA fibroblasts in an extensive fibrotic area. WNT signaling-activated (s/abeta-catenin-expressing) synovial fibroblasts upregulated WISP2 at 2.9-fold, but -inactivated (Deltabeta catenin-expressing) cells downregulated the expression. Quantitative RT-PCR demonstrated that WISP2 expression was increased upon 17-beta-estradiol stimulation and synergistically enhanced by WNT signaling. These data demonstrate that the expression of WISP2 is synergistically upregulated in RA synovial fibroblasts by estrogen and WNT pathways, and suggest an involvement in the pathology of the disease. PMID- 16038876 TI - Serotonin transporter polymorphisms affect human blood glucose control. AB - We measured the effect of nutritional intervention on clinical data, including fasting blood glucose (FBG), and their association with polymorphisms of the serotonin transporter-linked polymorphic region (5-HTTLPR) which might affect adherence. Enrolled in the intervention program were 264 Japanese women not on medication for diabetes, hypercholesterolemia or hypertension. The 5-HTTLPR allele (S and L) frequencies among the subjects differed markedly from those of Caucasians: SS (n = 183), LS (n = 69), and LL (n = 12). The decrease in FBG (DeltaFBG) from the beginning to the end of the program (11 weeks; short-term study), and DeltaFBG from the beginning to a follow-up check performed between 2002 and 2004 (average of 23 years later; long-term study) was calculated. The SS homozygotes of 5-HTTLPR showed larger DeltaFBG (P = 0.01 and P < 0.0001 in the short- and long-term studies, respectively) than DeltaFBG with other genotypes. PMID- 16038877 TI - Early molecular-level changes in rat bladder wall tissue following spinal cord injury. AB - Previously, we demonstrated using a rat model of spinal cord injury (SCI) that bladder wall tissue compliance significantly increased within the first 2 weeks following injury. In order to explore the potential molecular-level mechanisms of this event, the present study quantified molecules pertinent to bladder tissue remodeling and changes in mechanical properties. An initial gene array analysis followed by real-time qPCR revealed that the message levels for tropoelastin and lysyl oxidase were as high as 8-fold in SCI rats compared to normal. Furthermore, both the message and protein levels of TGF-beta1 and IGF-1, known stimulators of elastin synthesis, in SCI rat bladders were significantly higher compared to those of normal rats. Taken together, it can be speculated that functional changes of the bladder associated with SCI induce release of select growth factors, which, in turn, stimulate elastogenesis that lead to alteration of biomechanical properties of the wall tissue. PMID- 16038878 TI - Neuronal sodium channels in ventricular heart cells are localized near T-tubules openings. AB - Cardiac voltage-dependent sodium channels (VDSC) are known to be tetrodotoxin (TTX)-resistant. However, recent immunochemical studies suggest the presence of TTX-sensitive neuronal-type VDSC in the heart. Scanning ion conductance microscopy (SICM) coupled to electrophysiology was used to obtain more direct functional evidence. TTX sensitivities of whole-cell sodium currents (I(Na)) in control and detubulated cells were compared. Addition of 200 nM TTX decreased I(Na) of control cells by 20%, whereas detubulated cells were hardly effected. The remaining current peaked slightly earlier and inactivation decay was faster (as in neuronal VDSC) than in detubulated cells. Single-channel activity was first assayed at random on the plasmalemma, and after topography had been revealed by SICM, at patched T-tubules openings. In the latter case, a single channel conductance of 11-12pS was observed with a higher rate of success. This study provides independent evidence for neuronal VDSC in cardiomyocytes where they could rapidly and synchronously couple T-tubule and cell surface depolarizations. PMID- 16038879 TI - Superoxide targets calcineurin signaling in vascular endothelium. AB - Superoxide emerges as key regulatory molecule in many aspects of vascular physiology and disease, but identification of superoxide targets in the vasculature remains elusive. In this work, we investigated the possibility of inhibition of protein phosphatase calcineurin by superoxide in endothelial cells. We employed a redox cycler 2,3-dimethoxy-1,4-naphthoquinone (DMNQ) to generate superoxide inside the cells. DMNQ caused inhibition of cellular calcineurin phosphatase activity, which was reversible upon DMNQ removal. Inhibition was suppressed by pre-incubating the cells with copper/zinc superoxide dismutase (Cu,ZnSOD). In addition, reducing cellular Cu,ZnSOD activity by diethylthiocarbamic acid treatment resulted in calcineurin inhibition and enhanced sensitivity to DMNQ. Further, we could show that DMNQ inhibits calcineurin-dependent nuclear translocation and transcriptional activation of NFAT transcription factor, and Cu,ZnSOD or superoxide scavenger Tiron reduced the inhibition. Thus, superoxide generation in endothelial cells results in inhibition of calcineurin signaling, which could have important pathophysiological implications in the vasculature. PMID- 16038880 TI - Characterization of new D-beta-aspartate-containing proteins in a lens-derived cell line. AB - Although proteins are generally composed of l-alpha-amino acids, biologically uncommon D-beta-aspartic acid (Asp)-containing proteins have been reported in various tissues from elderly individuals. Our previous study indicated that the N/N1003A cell line, derived from rabbit lens, includes D-beta-Asp-containing proteins of approximately 50 kDa by Western blot analysis of a 2D-gel using a polyclonal antibody that is highly specific for D-beta-Asp-containing proteins. In this study, we identified the D-beta-Asp-containing proteins by matrix assisted laser desorption/ionization time-of-flight mass spectrometry and the Mascot online database searching algorithm. The results indicate that one of these 50 kDa proteins is an enolase showing homology with tau-crystallin. Other D beta-Asp-containing proteins, which we have recently discovered include lamin A/C, cytoplasmic NADP+-dependent isocitrate dehydrogenase, fructose-bisphosphate aldolase A, aldose reductase, L-lactate dehydrogenase A or calponin H2, phosphoglycerate mutase 1, phosphatidylethanolamine-binding protein, alpha-B crystallin, and peptidyl-prolyl cis-trans isomerase A (PPlase). PMID- 16038881 TI - Demonstration of an in vivo generated sub-picomolar affinity fully human monoclonal antibody to interleukin-8. AB - The high specificity and affinity of monoclonal antibodies make them attractive as therapeutic agents. In general, the affinities of antibodies reported to be high affinity are in the high picomolar to low nanomolar range and have been affinity matured in vitro. It has been proposed that there is an in vivo affinity ceiling at 100 pM and that B cells producing antibodies with affinities for antigen above the estimated ceiling would have no selective advantage in antigen induced affinity maturation during normal immune responses. Using a transgenic mouse producing fully human antibodies, we have routinely generated antibodies with sub-nanomolar affinities, have frequently rescued antibodies with less than 10 pM affinity, and now describe the existence of an in vivo generated anti-hIL-8 antibody with a sub-picomolar equilibrium dissociation constant. This confirms the prediction that antibodies with affinities beyond the proposed affinity ceiling can be generated in vivo. We also describe the technical challenges of determining such high affinities. To further understand the importance of affinity for therapy, we have constructed a mathematical model to predict the relationship between the affinity of an antibody and its in vivo potency using IL 8 as a model antigen. PMID- 16038882 TI - Salicylic acid modulates oxidative stress and glutathione peroxidase activity in the rat colon. AB - Oxidative stress is a characteristic of cancerous colon tissue and inflammatory bowel diseases that increase colon cancer risk. Epidemiological evidence supports a protective effect of plant-derived compounds. Aspirin is also protective against colon cancer. The mechanism of action is unclear although salicylic acid, the main metabolite of aspirin, has been shown to decrease the synthesis of pro inflammatory and potentially neo-plastic prostaglandins. Salicylic acid is found in significant quantities in a plant-based diet. However, in plants salicylic acid is also reported to modulate the expression of numerous enzymes with antioxidant activity. The aim of this study was to assess whether salicylic acid can modulate pro-cancerous biological pathways in the colon. Oxidative stress, prostaglandins and cytosolic glutathione peroxidase (cyGPX) were analysed in proximal, transverse and distal colon from a rat model of diet-induced oxidative stress. Elevated plasma pyruvate kinase activity (1293+/-206 U/ml) and increased indices of lipid peroxidation in colon (proximal 6.4+/-0.84 nM MDA/mg protein; transverse 6.9+/-0.97 nM MDA/mg protein; distal 5.2+/-0.62 nM MDA/mg protein) from rats fed a Vitamin E deficient diet were significantly decreased on supplementation with salicylic acid (plasma pyruvate 546+/-43 U/ml; salicylic acid proximal 3.6+/-0.39 nM MDA/mg protein; transverse 4.5+/-0.61 nM MDA/mg protein; distal 4.4+/-0.27 nM MDA/mg protein). Reductions in oxidative stress and prostaglandin production on supplementation with salicylic acid were associated with an elevation in glutathione peroxidase activity (Vitamin E deficient proximal 0.056+/-0.013 U/mg protein; transverse 0.073+/-0.008 U/mg protein; distal 0.088+/-0.010 U/mg protein; Vitamin E deficient with salicylic acid proximal 0.17+/-0.01 U/mg protein; transverse 0.23+/-0.016 U/mg protein; distal 0.16+/-0.020 U/mg protein). Gpx1 and Gpx2 gene transcripts were not elevated in association with increased activity of the soluble glutathione peroxidase activity. Glutathione peroxidases are key antioxidant enzymes, catalysing the decomposition of potentially toxic lipid peroxides. Gpx activity and regulation of Gpx gene transcription has been shown previously to be complex with activity not necessarily mirrored by a corresponding elevation in gene transcription. By supplementing the diet of Vitamin E deficient rats with salicylic acid (1 g/kg diet), this study assessed effects of salicylic acid on cytosolic glutathione peroxidase activity in the colon. The ability of salicylic acid to modulate antioxidant enzymes in colon tissue may be an important mechanism in inhibiting colon cancer development. PMID- 16038883 TI - Involvement of NMDA receptors in the hypotensive response to the injection of L glutamate into the lateral hypothalamus of unanesthetized rats. AB - We report that microinjections of L-glutamate (L-glu) or N-methyl-D-aspartic acid (NMDA) in the lateral hypothalamus (LH) of unanesthetized rats caused a hypotensive response. Guide cannulas were stereotaxically placed in the LH 3 days before the experiments, under tribromoethanol anesthesia. One day before the experiments, the femoral artery was cannulated for pulsatile arterial pressure (PAP), mean arterial pressure (MAP) and heart rate (HR) measurements. In the first experiment, unanesthetized rats received microinjections of 2.5, 5.0 or 10.0 nmol/100 nL of L-glu in the LH. Dose-dependent hypotensive responses were observed, without significant concomitant changes in heart rate. In a second group of experiments, 5.0 nmol of L-glu was microinjected into the LH before and 10 min after pretreatment with glutamatergic antagonists. Pretreatments with the non-selective ionotropic glutamatergic-receptor antagonist kynurenic acid or the selective NMDA receptor antagonists AP-7 and LY235959 significantly reduced the hypotensive response to microinjection of L-glu in the LH. Pretreatment with the selective AMPA-receptor antagonist NBQX or with vehicle did not affect the hypotensive response. The present results suggest that the hypotensive response to the injection of L-glu into the LH is mediated by NMDA receptors. PMID- 16038884 TI - Cerebral ischemia induced apoptosis and necrosis in normal and diabetic rats. AB - Stroke is the third leading cause of death and disability, and the risk for ischemic stroke is greater in diabetics. Previous studies have demonstrated both structural and functional nervous system changes in diabetes, and these changes may be enhanced by apoptosis. In the present study, we evaluated several indexes of both necrosis and apoptosis in the CNS of normals and two different models of diabetes (insulinopenic and insulin-resistant). Studies were conducted following middle cerebral artery occlusion (MCAO) with or without reperfusion. The sensory motor cortex (layer-5 and -6) and the CA1 and CA3 sectors of the hippocampus were analyzed following MCAO. We observed that both insulinopenic and insulin resistant diabetic rats have increased basal level of apoptosis that is uniformly and bilaterally distributed as indicated by both caspase-3 activity and TUNEL staining. Twenty-four hours after MCAO, apoptosis was further increased in both diabetic models. Reperfusion after a 2 h MCAO compared to 24 h MCAO was associated with a decrease in TUNEL staining and caspase-3 activity in the control animal but exacerbated apoptosis, especially in the hippocampus of insulin-resistant diabetic rats. MCAO-induced lesion volumes were greater in insulinopenic rats compared to insulin-resistant and control rats. We conclude that both insulinopenic and insulin-resistant diabetic animals have increased apoptosis in the CNS in response to MCAO, and restoration of blood flow especially in the insulinopenic diabetic animals paradoxically exacerbates this process. Furthermore, restoration of blood flow did not decrease lesion volume in insulinopenic diabetic animals. PMID- 16038885 TI - Differential co-localization of neurokinin-3 receptor and NMDA/AMPA receptor subunits in neurons of the substantia nigra of C57/BL mice. AB - By using a double immunofluorescence method we examined co-localization of neurokinin-3 receptor (NK-3R) and N-methyl-D-aspartate (NMDA)/alpha-amino-3 hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptor subunits in neurons of the substantia nigra of adult mice. Overlapping distribution of NMDA receptor subunit 1 (NR1)/AMPA receptor subunits 1-4 (GluR1-4) and NK-3R-immunoreactive neurons were found in the substantia nigra pars compacta. It revealed that all (100%) of NK-3R-positive neurons displayed NR1, GluR2 or GluR3 immunoreactivity, 80% of them showed GluR1 immunoreactivity. In contrast, these neurons exhibiting both NK-3R and GluR4 immunoreactivity were hardly detected although GluR4 positive neurons were still distributed in the substantia nigra. The co expression of NK-3R and NMDA/AMPA receptor subunits in the nigral neurons has provided a structural basis for functional modulation of neuronal glutamate receptors by neurokinin-3, suggesting that neurokinin peptides may be involved in modulation of neuronal properties and excitotoxicity in the substantia nigra of basal ganglia. PMID- 16038886 TI - Glutamate and GABA immunocytochemical electron microscopy in the hippocampal dentate gyrus of normal and genetic absence epilepsy rats. AB - It is generally accepted that absence epilepsy results from the impairment of GABAergic and glutamatergic neurotransmission. In particular, besides excessive GABA mediation within the thalamo-cortico-thalamic circuit in absence epilepsy, neuronal networks of the hippocampus have recently received attention. In the present study, we examined the density of glutamate and GABA neurotransmitter immunolabeling in the dentate gyrus of the hippocampus of genetic absence epilepsy rats from Strasbourg (GAERS) compared to the control group. GABA and glutamate were found to exist in synaptic vesicles of the mossy fiber terminals of the control and GAERS groups. The density of glutamate immunolabeling within the mossy fiber terminals in the hilar region of GAERS hippocampus was found to be significantly decreased compared to the control group. There was no difference in the density of immunolabeling within GABA nerve terminals between GAERS and control group. The findings of this study suggest that mechanisms underlying absence seizures in GAERS may also manifest themselves in other brain regions such as the hippocampus. The presence of GABA within synaptic vesicles of mossy fiber terminals, as revealed by high resolution ultrastructural immunocytochemistry, has provided additional evidence to the possible modulatory role of GABA on synaptic transmission between the mossy fiber and the target cell. PMID- 16038887 TI - The effect of short duration streptozotocin-induced diabetes mellitus on the late phase and threshold of long-term potentiation induction in the rat. AB - Long-term potentiation (LTP) was examined in streptozotocin (STZ)-induced diabetic rats of 8 (DM8) and 20 (DM20) weeks duration of diabetes mellitus (DM). DM8 animals showed significant LTP induction, although the potentiation of the synapses was significantly lower than in the control animals. No significant potentiation of the synapses could be demonstrated in DM20 animals. The different aspects of LTP induction in the DM8 animals were studied. The threshold of LTP induction was measured by stimulating the slices with 100 Hz frequency trains of stimuli containing different number of impulses. The results showed increased threshold for LTP induction in the DM8 animals compared to the controls. The late LTP (L-LTP) phase induction was studied by applying 3 repeated HFSs to the afferent fibers. Diabetic animals (DM8) slices failed to maintain the synaptic potentiation induced by the high frequency stimulations (HFSs) for more than 1 h. PMID- 16038889 TI - A 'hook' attached to the fish-mouth technique for tendon repair. PMID- 16038890 TI - QM/MM study of D-fructose in aqueous solution. AB - The QM/MM molecular dynamics methodology was applied to the study of the two main D-fructose tautomers present in aqueous solution, beta-D-fructofuranose and beta D-fructopyranose. The solute was treated at the AM1 semi-empirical level, and for the solvent water molecules we used the TIP3P potential. We analyzed the structure of the water molecules around the hydroxyl groups to explain the differences in sweet taste between the two tautomers. PMID- 16038891 TI - Substantial proportions of identical beta-chain T-cell receptor transcripts are present in epithelial ovarian carcinoma tumors. AB - To determine whether clonally expanded T cells are present in tumor specimens from patients with epithelial ovarian carcinoma (EOC) we amplified by the non palindromic adaptor PCR (NPA-PCR) or by Vbeta-specific PCR beta-chain T-cell receptor (TCR) transcripts from these tumor specimens. The amplified transcripts were cloned and sequenced. Sequence analysis revealed the presence of substantial proportions of multiple identical copies of beta-chain TCR transcripts, suggesting the presence of clonal expansions of T cells in these patients, which were statistically significant by the binomial distribution in seven of nine patients. Independent amplification in separate experiments of beta-chain TCR transcripts from one patient by either NPA-PCR or by Vbeta-specific PCR, followed by cloning and sequencing, revealed identical clonal expansions irrespectively of the amplification method used. Multiple identical copies of beta-chain TCR transcripts can be derived only by specific antigen-driven proliferation and clonal expansion of the T-cell clones which recognize these antigens. Because of the very large size of the TCR repertoire, the probability of finding by chance multiple identical copies of these transcripts within an independent sample of T cells is negligible. These results demonstrate that T cells infiltrating solid tumor specimens or malignant ascites of patients with EOC contain monoclonal/oligoclonal populations of T cells. PMID- 16038892 TI - Pharmacogenetic study of apolipoprotein E, cholesteryl ester transfer protein and hepatic lipase genes and simvastatin therapy in Brazilian subjects. AB - BACKGROUND: In recent years, one of the focuses of genetic investigation in cardiology has been to identify the genetic factors associated with variable response to statin treatment. Polymorphisms in apolipoprotein E (APOE), cholesteryl ester transfer protein (CETP) and hepatic lipase (LIPC), proteins with major roles in lipid metabolism and homeostasis have been shown associated with lipid-lowering drugs response. METHODS: One hundred forty-six hypercholesterolemic patients of European descent were prospectively enrolled and treated with simvastatin 20 mg per day for over 6 months. Ninety-nine subjects completed the 6-month follow-up. Plasma lipids and lipoproteins were measured before and throughout the study. APOE (E*2, E*3 and E*4), LIPC-250A > G and CETP TaqIB genotypes were determined by PCR and restriction mapping. RESULTS: After a 6-month follow-up, no differences among genotypes in the percentage variation in lipid and lipoprotein concentrations for APOE and LIPC SNPs were observed. After adjustment for covariates, CETP B2B2 homozygotes showed a greater HDL-cholesterol increase compared to B1B2 and B1B1 subjects (14.1% vs. 1.7% and 1.3%, P < 0.05, respectively). CONCLUSION: Our study demonstrates that individual plasma HDL cholesterol response to simvastatin is mediated, in part, by the CETP gene locus, with the B2 homozygotes having more benefit in HDL-C improvement than carriers of B1 allele. PMID- 16038894 TI - Pedicel development in Arabidopsis thaliana: contribution of vascular positioning and the role of the BREVIPEDICELLUS and ERECTA genes. AB - Although the regulation of Arabidopsis floral meristem patterning and determinacy has been studied in detail, very little is known about the genetic mechanisms directing development of the pedicel, the short stem linking the flower to the inflorescence axis. Here, we provide evidence that the pedicel consists of a proximal portion derived from the young flower primordium, and a bulged distal region that emerges from tissue at the bases of sepals in the floral bud. Distal pedicel growth is controlled by the KNOTTED1-like homeobox gene BREVIPEDICELLUS (BP), as 35S::BP plants show excessive proliferation of pedicel tissue, while loss of BP conditions a radial constriction around the distal pedicel circumference. Mutant radial constrictions project proximally along abaxial and lateral sides of pedicels, leading to occasional downward bending at the distal pedicel. This effect is severely enhanced in a loss-of-function erecta (er) background, resulting in radially constricted tissue along the entire abaxial side of pedicels and downward-oriented flowers and fruit. Analysis of pedicel vascular patterns revealed biasing of vasculature towards the abaxial side, consistent with a role for BP and ER in regulating a vascular-borne growth inhibitory signal. BP expression in a reporter line marked boundaries between the inflorescence stem and lateral organs and the receptacle and floral organs. This boundary expression appears to be important to prevent homeotic displacement of node and lateral organ fates into underlying stem tissue. To investigate interactions between pedicel and flower development, we crossed bp er into various floral mutant backgrounds. Formation of laterally-oriented bends in bp lfy er pedicels paralleled phyllotaxy changes, consistent with a model where the architecture of mutant stems is controlled by both organ positioning and vasculature patterns. Collectively, our results indicate that the BP gene acts in Arabidopsis stems to confer a growth-competent state that counteracts lateral organ associated asymmetries and effectively radializes internode and pedicel growth and differentiation patterns. PMID- 16038893 TI - Multiple requirements for Hes 1 during early eye formation. AB - During embryogenesis, multiple developmental processes are integrated through their precise temporal regulation. Hes1 is a transcriptional repressor that regulates the timing of mammalian retinal neurogenesis. However, roles for Hes1 in early eye development have not been well defined. Here, we show that Hes1 is expressed in the forming lens, optic vesicle, cup, and pigmented epithelium and is necessary for proper growth, morphogenesis, and differentiation of these tissues. Because Hes1 is required throughout the eye, we investigated its interaction with Pax6. Hes1-Pax6 double mutant embryos are eyeless suggesting these genes are coordinately required for initial morphogenesis and outgrowth of the optic vesicle. In Hes1 mutants, Math5 expression is precocious along with retinal ganglion cell, amacrine, and horizontal neuron formation. In contrast to apparent cooperativity between Pax6 and Hes1 during morphogenesis, each gene regulates Math5 and RGC genesis independently. Together, these studies demonstrate that Hes1, like Pax6, simultaneously regulates multiple developmental processes during optic development. PMID- 16038895 TI - Short-type PB-cadherin promotes survival of gonocytes and activates JAK-STAT signalling. AB - Neonatal development of the rat testis involves a number of critical events including re-entry of gonocytes into the cell cycle and eventual loss of many of these cells and their progeny via apoptosis. Since surviving gonocytes give rise to subsequent generations of germ cells, regulation of their fate is critical for adult testicular function. Here, we have identified a role for short-type PB cadherin (STPB-C) in promoting survival of gonocytes in neonatal rats and we have linked its expression to the JAK-STAT signaling pathway. These findings were obtained with varied approaches including use of transgenic rats overexpressing STPB-C which were studied with protein microarrays and other techniques, direct examination of germ cell apoptosis and survival in gonocyte-Sertoli cell co cultures, and direct study of the JAK-STAT pathway in these models and in L cells transfected with STPB-C. These data provide new information on the regulation of gonocyte fate and exciting new evidence supporting a link between the JAK-STAT pathway and cadherin-based cell-cell interactions. PMID- 16038896 TI - Tandem mass spectrometry identifies proteins phosphorylated by cyclic AMP dependent protein kinase when sea urchin sperm undergo the acrosome reaction. AB - The exocytotic acrosome reaction (AR), which is required for fertilization, occurs when sea urchin sperm contact the egg jelly (EJ) layer. Among other physiological changes, increases in adenylyl cyclase activity, cAMP and cAMP dependent protein kinase (PKA) activity occur coincident with the AR. By using inhibitors of PKA, a permeable analog of cAMP and the phosphodiesterase inhibitor IBMX, we show that PKA activity is required for AR induction by EJ. A minimum of six sperm proteins are phosphorylated by PKA upon exposure to EJ, as detected by a PKA substrate-specific antibody. The phosphorylation of these proteins and the percentage of acrosome reacted sperm can be regulated by PKA modulators. The fucose sulfate polymer (FSP), a major component of EJ, is the molecule that triggers sperm PKA activation. Extracellular Ca(2+) is required for PKA activation. Six sperm proteins phosphorylated by PKA were identified by tandem mass spectrometry (MS/MS) utilizing the emerging sea urchin genome. Based on their identities and localizations in sperm head and flagellum, the putative functions of these proteins in sperm physiology and AR induction are discussed. PMID- 16038897 TI - Encepalomyocarditis virus-induced apoptosis and ultrastructural changes in the lacrimal and parotid glands of mice. AB - Development of acinar cell apoptosis and ultrastructural changes in the exorbital lacrimal and parotid glands was examined in DBA/2 mice infected with 10(2) PFU/mouse of EMC-D virus. Pyknotic acinar cells, most of which were positive for TUNEL and cleaved caspase-3 and had ultrastructural characteristics of apoptotic cells, developed earlier and were more frequently observed in the parotid gland than in the exorbital lacrimal gland, while the total damage of acinar cells and interstitial infiltration of macrophages were more prominent in the latter than in the former. These findings indicate that EMC-D virus induces acinar cell apoptosis in these glands. In addition, corresponding to the results of the detection of viral RNA signals by in situ hybridization, small aggregates of virus-like particles having typical size and structure of EMC virus were frequently observed in both the cytoplasm and the nucleus of acinar cells in the exorbital lacrimal gland, while they were found only in the cytoplasm of a few acinar cells in the parotid gland. In conclusion, between the exorbital lacrimal and parotid glands, there was a reverse relationship observed between the development of acinar cell apoptosis and that of total damage of acinar cells. PMID- 16038898 TI - Clusterin interacts with SCLIP (SCG10-like protein) and promotes neurite outgrowth of PC12 cells. AB - Clusterin has been known as a chaperone-like molecule capable of interacting with various proteins. In this study, we show that clusterin interacts with the microtubule-destabilizing stathmin family protein SCLIP by GST pull-down and co immunoprecipitation assays. Interestingly, SCLIP interacts with 80 kDa mature form of clusterin in the cytosolic fraction of PC12 cells permeabilized by low concentration of a weak nonionic detergent digitonin, but not with intracellular variants of clusterin known as binding isoforms of Ku70 or TGF-beta receptors. Both clusterin and SCLIP are co-localized at the perinuclear region and growth cone of PC12 cells. In addition, we show that the minimal domains for the interaction are mapped to the C-terminal valine-rich region (367-447) of clusterin and the N-terminal palmitoylation and membrane attachment site (1-34) of SCLIP. Finally, we demonstrate that ectopic expression of clusterin in PC12 cells elongates neurite-formation triggered by NGF and induces spontaneous neurite outgrowth even in the absence of NGF. Taken together, these results suggest that the clusterin interacts with SCLIP and the interaction may act as an important modulator during neuronal differentiation. PMID- 16038899 TI - Monomeric and oligomeric flavanols are agonists of membrane androgen receptors. AB - The present work reports a new mode of action of the naturally occurring flavanols catechin and epicatechin and their dimers B2 and B5, in the breast cancer T47D cell line, namely, their interaction with membrane androgen receptors. We show that monomeric and dimeric flavanols are complete (B2) or partial displacers of radiolabeled testosterone bound on T47D membranes, with affinities ranging from 1.7 (B5) to 82.2 nM (B2). In addition, they trigger the phosphorylation of the same signaling molecules (FAK, PI3K) as testosterone-BSA, minutes after binding to membrane receptors, leading to actin cytoskeleton polymerization and redistribution, with formation of filopodia and lamellipodia. The PI3K inhibitor wortmannin reverts the effect of polyphenols and testosterone BSA, providing additional evidence about activation of a similar signaling cascade. Incubation of T47D cells for more than 2 h with polyphenols or testosterone-BSA induces apoptosis, which follows the same time-dependent pattern. We conclude that flavanols (monomers or dimers) are agonists of membrane androgen receptors and could be used as testosterone-protein conjugates for the management of tumors, in which, application of testosterone-BSA induces regression, providing additional data about the mechanism of their antiproliferative action. PMID- 16038900 TI - Focus on molecules: the Sigma-1 receptor. PMID- 16038901 TI - Trypanosoma cruzi: influence of predominant bacteria from indigenous digestive microbiota on experimental infection in mice. AB - To verify the influence of some predominant components from indigenous microbiota on systemic immunological responses during experimental Chagas disease, germ-free NIH Swiss mice were mono-associated with Escherichia coli, Enterococcus faecalis, Bacteroides vulgatus or Peptostreptococcus sp. and then infected with the Y strain of Trypanosoma cruzi. All the mono-associations predominantly induced a Th1 type of specific immune response to the infection by T. cruzi. A direct correlation was observed between a higher survival rate and increased IFN-gamma and TNF-alpha production (P<0.05) in E. faecalis-, B. vulgatus-, and Peptostreptococcus-associated mice. Moreover, higher levels of anti-T. cruzi IgG1 and anti-T. cruzi IgG2a were also found in mono-associated animals after infection. On the other hand, with the exception of E. faecalis-associated mice, mono-association induced a lower IL-10 production after infection (P<0.05) when compared with germ-free animals. Interestingly, spleen cell cultures from non infected germ-free and mono-associated mice spontaneously produced higher levels (P<0.05) of IL-10 than cultures from infected mono-associated mice, except again for E. faecalis-associated animals. In conclusion, the presence of the components of the indigenous microbiota skews the immune response towards production of inflammatory cytokines during experimental infection with T. cruzi in gnotobiotic mice. However, the degree of increase in production of cytokines depends on each bacterial component. PMID- 16038902 TI - Coordinated synthesis of the two ClpB isoforms improves the ability of Escherichia coli to survive thermal stress. AB - Eubacteria synthesize a full-length (ClpB95) and a N-terminally truncated (ClpB80) version of the ClpB disaggregase owing to the presence of a translation initiation site within the clpB transcript. Why these two isoforms have been evolutionary conserved is poorly understood. Here, we constructed a series of E. coli strains and plasmids allowing production of the ClpB95/ClpB80 pair, ClpB95 alone, or ClpB80 alone from near physiological concentrations to a 6-10-fold excess over normal cellular levels. We found that although overexpressed ClpB95 or ClpB80 can independently restore basal thermotolerance to DeltaclpB cells, strains expressing ClpB80 from the clpB chromosomal locus do not exhibit increased resistance to thermal killing at 50 degrees C relative to clpB null cells. Furthermore, synthesis of physiological levels of ClpB95 is less effective than coordinated expression of ClpB95/ClpB80 in protecting E. coli from thermal killing. These results provide an explanation for the conservation of the two ClpB isoforms in eubacteria and are consistent with the fact that wild type E. coli maintains the ClpB80 to ClpB95 ratio at a nearly constant value of 0.4-0.5 under a variety of stress conditions. PMID- 16038903 TI - Nucleotide binding to the homodimeric MJ0796 protein: a computational study of a prokaryotic ABC transporter NBD dimer. AB - Transport by ABC proteins requires a cycle of ATP-driven conformational changes of the nucleotide binding domains (NBDs). We compare three molecular dynamics simulations of dimeric MJ0796: with ATP was present at both NBDs; with ATP at one NBD but ADP at the other; and without any bound ATP. In the simulation with ATP present at both NBDs, the dimeric protein interacts with the nucleotides in a symmetrical manner. However, if ADP is present at one binding site then both NBD NBD and protein-ATP interactions are enhanced at the opposite site. PMID- 16038904 TI - The co-chaperone p23 is degraded by caspases and the proteasome during apoptosis. AB - The heat shock protein 90 co-chaperone p23 has recently been shown to be up regulated in cancer cells and down-regulated in atheroschlerotic plaques. We found that p23 is degraded during apoptosis induced by several stimuli, including Fas and TNFalpha-receptor activation as well as staurosporine treatment. Caspase inhibition protected p23 from degradation in several cell lines. In addition, recombinant caspase-3 and 8 cleaved p23 at Asp 142 generating a degradation product of 18 kDa as seen in apoptotic cells. Truncated p23 is further degraded in a proteasome dependent process during apoptosis. Furthermore, we found that the anti-aggregating activity of truncated p23 was reduced compared to full length p23 indicating that caspase mediated p23 degradation contributes to protein destabilisation in apoptosis. PMID- 16038905 TI - OD1, the first toxin isolated from the venom of the scorpion Odonthobuthus doriae active on voltage-gated Na+ channels. AB - In this study, we isolated and pharmacologically characterized the first alpha like toxin from the venom of the scarcely studied Iranian scorpion Odonthobuthus doriae. The toxin was termed OD1 and its primary sequence was determined: GVRDAYIADDKNCVYTCASNGYCNTECTKNGAESGYCQWIGRYGNACWCIKLPDEVPIRIPGKCR. Using the two electrode voltage clamp technique, the pharmacological effects of OD1 were studied on three cloned voltage-gated Na+ channels expressed in Xenopus laevis oocytes (Na(v)1.2/beta1, Na(v)1.5/beta1, para/tipE). The inactivation process of the insect channel, para/tipE, was severely hampered by 200 nM of OD1 (EC50 = 80+/-14 nM) while Na(v)1.2/beta1 still was not affected at concentrations up to 5 microM. Na(v)1.5/beta1 was influenced at micromolar concentrations. PMID- 16038906 TI - Probing cytoskeleton modulation by optical biosensors. AB - This paper reported the use of resonant waveguide grating biosensors for studying the cytoskeleton structure in cells. This was achieved by measuring the changes in mass within the bottom portion of cells upon exposure to saponin in the absence and presence of cytoskeleton modulators. Treatment of Chinese hamster ovary cells with saponin led to a dose-dependent and dynamic mass changes. When a higher concentration of saponin (> 60 microg/ml) was used, a net loss in mass was observed. This is probably resulted from the diffusion of soluble intracellular materials away from the bottom portion of cells after pore formation in the cell plasma membranes by saponin. The pretreatment of cells with actin disruption agents, cytochalasin B and latrunculin A, led to significantly increased loss in cell mass induced by either 75 or 125 microg/ml saponin. These results suggested that optical biosensors provide an attractive means to study the cytoskeleton structure and screen modulators that affect the cytoskeleton structure. PMID- 16038907 TI - Arabidopsis thaliana MTP1 is a Zn transporter in the vacuolar membrane which mediates Zn detoxification and drives leaf Zn accumulation. AB - The Arabidopsis thaliana metal tolerance protein 1 (MTP1) of the cation diffusion facilitator family of membrane transport proteins can mediate the detoxification of Zn in Arabidopsis and yeast. Xenopus laevis oocytes expressing AtMTP1 accumulate more Zn than oocytes expressing the AtMTP1(D94A) mutant or water injected oocytes. An AtMTP1-GFP fusion protein localizes to the vacuolar membrane in root and leaf cells. The analysis of Arabidopsis transformed with a promoter GUS construct suggests that AtMTP1 is not produced throughout the plant, but primarily in the subpopulation of dividing, differentiating and expanding cells. RNA interference-mediated silencing of AtMTP1 causes Zn hypersensitivity and a reduction in Zn concentrations in vegetative plant tissues. PMID- 16038908 TI - Validated HPLC method for determination of amlodipine in human plasma and its application to pharmacokinetic studies. AB - A rapid, simple and sensitive high-performance liquid chromatography (HPLC) method has been developed for quantification of amlodipine in plasma. The assay enables the measurement of amlodipine for therapeutic drug monitoring with a minimum detectable limit of 0.2 ng ml(-1). The method involves simple, one-step extraction procedure and analytical recovery was about 97%. The separation was performed on an analytical 125 x 4.6 mm i.d. Nucleosil C8 column. The wavelength was set at 239 nm. The mobile phase was a mixture of 0.01 M sodium dihydrogen phosphate buffer and acetonitrile (63:37, v/v) adjusted to pH 3.5 at a flow rate of 1.5 ml min(-1). The calibration curve was linear over the concentration range 0.5-16 ng ml(-1). The coefficients of variation for inter-day and intra-day assay were found to be less than 10%. PMID- 16038909 TI - Synthesis and anti-HIV activity of novel phenyl branched cyclopropyl nucleosides. AB - Novel phenyl branched cyclopropyl nucleoside analogues were designed and synthesized as potential antiviral agents. Cyclopropanation was performed via classical Simmons-Smith reaction using Zn(Et)2 and CH2I2. Coupling of the mesylates 11 and 12 with natural bases (A,C,T,U) and desilylation afforded a series of novel cyclopropyl nucleosides 21-28. The synthesized compounds were evaluated for their antiviral and antitumor activity against various viruses such as HIV, HSV-1, HSV-2 and HCMV. PMID- 16038911 TI - Students' perceptions on the use of portfolios in pre-registration nursing education: a questionnaire survey. AB - BACKGROUND: Portfolios encourage both personal and professional development through the process of reflective practice and critical analysis. In clinical placements portfolios are used both as a learning tool and as an assessment tool. OBJECTIVES: To determine nursing students' perceptions on the use and effectiveness of portfolios in their education as well as their perceptions on the use of the portfolio as both an assessment and learning tool in clinical placement. DESIGN: Postal questionnaire survey. SETTINGS: Higher Education Institute in the UK. PARTICIPANTS: 90 first and 84 third year pre-registration diploma of nursing students. METHODS: Postal questionnaires were sent to 131 first year and 122 third year students, with a reminder 3 weeks later. The total response rate was 69% (174/253), with a response rate of 69% (90/131) for first year students and a response rate of 69% (84/122) for third year students. RESULTS: Students felt that portfolios were very time-consuming, causing them a great deal of anxiety, and were not very effective in developing and assessing their learning and competence. Using portfolios for both assessment and learning creates a conflict. Any assessment reduces the honesty and learning value of reflective writing and of the portfolio. Additionally, students become increasingly demoralized with portfolio use over time with experience. The study suggested three main linked reasons for this, namely the conflict between using portfolios for both assessment and learning, the design of the portfolio and the amount of support and guidance students feel they receive with their portfolio use. CONCLUSIONS: Portfolios can be very effective as an assessment and learning tool, but it is essential that both students and mentors receive clear guidelines on and comprehensive support with their use. They should be designed in such as way that they are relevant, clear and user-friendly for both students and mentors. PMID- 16038912 TI - Long-term achievement of the therapeutic objectives of lipid-lowering agents in primary prevention patients and cardiovascular outcomes: an observational study. AB - AIMS: Lowering elevated cholesterol levels reduces cardiovascular (CV) morbidity and mortality. Nonetheless, most patients treated with lipid-lowering agents (LLA) do not reach recommended therapeutic objectives. In a setting of primary care in France, we investigated the association between LDL-cholesterol goal attainment and the occurrence of CV events in primary prevention patients with multiple CV risk factors (> or = 3). According to national guidelines, the therapeutic objective (TO) for such patients is an LDL-cholesterol value below 130 mg/dL. METHODS: 579 patients treated with LLA and with LDL-cholesterol values documented at least once a year over a period of at least 3 years (2000-2002) were allocated to three groups based on the number of years the TO was attained during the follow-up period: in all 3 years (TO+++: n=145), only part of the time (TO intermediate: n=256), and never (TO---: n=178). CV events (angina pectoris, myocardial infarction, heart failure, stroke, peripheral artery disease) occurring during the last year of observation (2002) were retrospectively collected. The occurrence risk (OR) of CV events was assessed based on TO status, with a logistic regression model to adjust for baseline differences in CV risk factors. RESULTS: Only a quarter of patients attained TO during all 3 study years. CV events during the third year of observation occurred in 5.5%, 10.5% and 12.9% of patients in the TO+++, TO intermediate and TO--- groups, respectively. Compared with TO+++ patients, the risk of CV events increased significantly in TO intermediate (OR=2.34, 95% CI=[1.01-5.39]) and TO--- patients (OR=2.99, 95% CI=[1.26-7.08]). CONCLUSION: In real practice, a prolonged attainment of TO is rarely observed in high CV risk patients treated with LLA as primary prevention. Therapeutic failure is related to an increased incidence of cardiovascular morbidity. Our data strongly support the need to improve adherence to treatment guidelines to achieve effective cardiovascular prevention. PMID- 16038913 TI - A large-strain finite element formulation for biological tissues with application to mitral valve leaflet tissue mechanics. AB - This paper presents a finite element formulation suitable for large-strain modeling of biological tissues and uses this formulation to implement an accurate finite element model for mitral valve leaflet tissue. First, an experimentally derived strain energy function is obtained from literature. This function is implemented in finite elements using the mixed pressure-displacement formulation. A modification is made to aid in maintaining positive definiteness of the stiffness matrix at low strains. The numerical implementation is shown to be accurate in representing the analytical model of material behavior. The mixed formulation is useful for modeling of soft biological tissues in general, and the model presented here is applicable to finite element simulation of mitral valve mechanics. PMID- 16038914 TI - A hypothesis for the function of braking forces during running turns. AB - We examined the functional role of braking forces observed when humans execute turning maneuvers. Deceleration caused by braking forces contributes to changing the movement direction of the center of mass (COM) and maintaining constant velocity. We argue that braking forces also prevent over-rotation of the body about the vertical axis during maneuvers. We analyzed data from sidestep and crossover cuts at average initial running velocities of 3 m s(-1). Absent braking, lateral forces would result in body rotations 1.4-3 times the change in COM movement direction, causing the orientation of the body to be substantially mis-aligned with the direction of movement at the end of the step. A simple model based on the hypothesis that body rotation should match COM deflection can explain 70% of the variance in braking forces employed during running turns. PMID- 16038915 TI - Development of residual strains in human vertebral trabecular bone after prolonged static and cyclic loading at low load levels. AB - Development of irreversible residual strains in trabecular bone may be a mechanism by which age-related non-traumatic vertebral fractures occur. To investigate this concept, static and cyclic loading tests were conducted at low loading levels for cylindrical cores of cadaveric vertebral trabecular bone. Stresses were applied equivalent to elastic strains of either 750 or 1,500 microstrain. Creep strains were measured during the tests, which lasted for 125,000 seconds (about 35 h), and for an additional 125,000 seconds after complete unloading. Emphasis was placed on the residual strains that developed, defined as the strain remaining at the end of the unloading phase. The results indicated that appreciable residual strains did develop, and were similar for static and cyclic loading. Irrespective of the applied load levels and loading modes, the residual strains that remained after the unloading phase were similar in magnitude to the originally applied elastic strain. Extrapolation of the observed residual strains to full recovery indicated that the time that would be required for full recovery was over 20 times longer than the duration of the applied loads. These results indicate that human vertebral trabecular bone does not creep in a linear viscoelastic fashion at low stress levels, and that creep mechanisms dominate the residual strains regardless of the loading mode. Taken together, these findings support the concept that non-traumatic vertebral fractures may be related to long-term creep effects because the trabecular bone does not have sufficient time to recover mechanically from creep deformations accumulated by prolonged static or cyclic loading. PMID- 16038917 TI - Biomechanical analysis of the shear behaviour adjacent to an axially loaded implant. AB - Good mechanical fixation of an implant to the surrounding bone is important for its longevity, and is influenced by both biological and mechanical factors. This study parametrically evaluates the mechanics of the interface with a computationally efficient analytic structural model of the shear stress field and global shear stiffness of an axially loaded implant. The utility of the analytic model was first established by validating its assumptions with a case-specific finite element model. We then used the analytic model for a sensitivity analysis of the relationship between the pattern of tissue growth and shear properties of the interface for our previously reported loaded in vivo experimental micromotion device. The bone located directly at the implant surface was found to be the most effective site for increasing interface stiffness. This suggests that the implant surface is the most desirable site for bone growth, yet is also the most mechanically challenging environment due to its maximal shear stresses. Thus, these findings support the further investigation of osteo-conductive coatings and other biological stimuli to overcome the challenging mechanics, and to promote bone growth directly at the implant surface. The model also demonstrated that the mechanical contribution to the global implant shear stiffness of a commonly observed isolated sclerotic bone rim is very limited. The results of this sensitivity analysis agree with experimental studies with the micromotion device, and with clinical studies reporting good results with osteo-conductive coatings. PMID- 16038916 TI - Regulation of RANKL by biomechanical loading in fibrochondrocytes of meniscus. AB - OBJECTIVE: We sought to determine whether fibrochondrocytes from menisci express receptor activator of NF-kappaB (RANK), its ligand (RANKL), or osteoprotegerin (OPG) and, if so, whether their expression is modulated by dynamic mechanical loading under inflammatory and normal conditions. METHODS: Fibrochondrocytes from rat menisci were subjected to cyclic tensile strain (CTS) at various magnitudes and frequencies in the presence or absence of interleukin (IL)-1beta for up to 24 h. In order to determine whether a possible regulatory effect of mechanical loading on RANKL and its receptors under inflamed conditions is sustained, cells were stimulated with IL-1beta for 24 h while being subjected to CTS only for the initial 4 and 8h, respectively. Regulation of RANKL, RANK, and OPG expression and synthesis were determined by semiquantitative and real-time PCR, Western blotting, and immunofluorescence. RESULT: Fibrochondrocytes constitutively expressed low levels of RANKL and RANK but marked levels of OPG. IL-1beta upregulated expression and synthesis of RANKL and RANK significantly (p<0.05), whereas expression of OPG was unaffected following 4 and 24 h. When fibrochondrocytes were simultaneously subjected to CTS and IL-1beta, expression of RANKL and RANK was significantly (p<0.05) downregulated as compared to that of IL-1beta-stimulated unstretched cells. The inhibitory effect of CTS on the IL 1beta-induced upregulation of RANKL and RANK was sustained as well as magnitude and frequency dependent. CONCLUSIONS: Our study provides evidence that RANKL and its receptors are expressed in fibrochondrocytes from meniscus. These data also demonstrate that dynamic mechanical loading can modify the expression of RANKL and RANK in inflammatory conditions. PMID- 16038918 TI - The effect of chelating agent on the separation of Fe(III) and Ti(IV) from binary mixture solution by cation-exchange membrane. AB - The competitive transport of Fe(III) and Ti(IV) ions and the effect of chelating agents on separation from binary mixture solutions through charged polysulfone cation-exchange membrane (SA3S) has been studied under Donnan dialysis conditions. The amount of chelating agent was taken as an equimolar of Fe(III) ion in the feed phase. In this process, the membrane separated two electrolyte solutions: the feed solution, initially containing metal salts (Fe, Ti), or metal salts solution, containing a chelating agent, and the other side (receiver solution) being HCl solution. An external potential field is not applied. It was observed that the chelating agents affect the metal transport; the transport of Fe(III) is decreased and the transport of Ti(IV) is increased. PMID- 16038919 TI - The effect of active carbon on the reduction of concentrated nitric acid by HCOOH. AB - In nuclear industry, removal of nitric acid from solutions is required in the course of chemical separation and waste treatment procedure as well as in chemical conversion steps. The reduction of HNO3 by HCOOH to gaseous products such as nitrogen, nitrogen oxides, and carbon dioxide is an attractive way to accomplish the denitration. A typical problem for the denitration is the existence of the induction period. The induction period has been explained as the time necessary to increase the concentration of HNO2, which is an important reaction intermediate, above a threshold value. In this study, adsorption sites on the surface of active carbon were found to promote HNO2 formation and efficiently suppress the induction period. Induction time was shortened by increasing the amount of active carbon in the solution. When the solution contains 3 M HNO3 and 1 M HCOOH, 10 g/L of active carbon was enough to eliminate the induction period at 50 degrees C. The catalytic effect exhibited by active carbon was similar to that reported for Pt/SiO2. Therefore, on the surface of active carbon, there is a redox cycle, where HNO3 is reduced to HNO2 and then the oxidized surface site will be reduced by HCOOH. PMID- 16038920 TI - Two-dimensional Monte Carlo simulations of a colloidal dispersion composed of rod like ferromagnetic particles in the absence of an applied magnetic field. AB - Influences of the magnetic interaction between particles and the aspect ratio of particles on aggregate structures in a colloidal dispersion composed of rod-like ferromagnetic particles were investigated by means of the cluster-moving Monte Carlo method. The internal structures of the aggregates obtained in simulations were analyzed in terms of the number density distribution of the clusters and radial distribution functions. The results show that as the magnetic interaction between particles increases, many small clusters such as anti-parallel particle pairs, raft-like clusters, and small loop-like clusters are formed; these gather to form larger aggregates. In the case of a relatively strong magnetic interaction between particles, solid-like rectangular clusters are formed when the aspect ratio is approximately 2, since the suitable distance between magnetic charges enables particles to form a fundamental structure of two normal anti parallel particle pairs. As the aspect ratio increases beyond 2, many more stable raft-like clusters are formed, since the increase in distance between magnetic charges makes the two normal anti-parallel particle pair structures unstable. For a significantly larger aspect ratio, large network microstructures are produced by the formation of many chain-like and loop-like structures. PMID- 16038921 TI - Improved dewatering behavior of clay minerals dispersions via interfacial chemistry and particle interactions optimization. AB - Orthokinetic flocculation of clay dispersions at pH 7.5 and 22 degrees C has been investigated to determine the influence of interfacial chemistry and shear on dewatering and particle interactions behavior. Modification of pulp chemistry and behavior was achieved by using kaolinite and Na-exchanged (swelling) smectite clay minerals, divalent metal ions (Ca(II), Mn(II)) as coagulants and anionic polyacrylamide copolymer (PAM A) and non-ionic polyacrylamide homopolymer (PAM N) as flocculants. The pivotal role of shear, provided by a two-blade paddle impeller, was probed as a function of agitation rate (100-500 rpm) and time (15/60 s). Particle zeta potential and adsorption isotherms were measured to quantify the interfacial chemistry, whilst rheology and cryogenic SEM were used to investigate particle interactions and floc structure and aggregate network, respectively. Osmotic swelling, accompanied by the formation of "honeycomb" particle network structure and high yield stress, was produced by the Na exchanged smectite, but not kaolinite, dispersions. Dispersion of the clay particles in 0.05 M Ca(II) or Mn(II) solution led to a marked reduction in particle zeta potential, complete suppression of swelling, honeycomb network structure collapse and a concomitant reduction in shear yield stress of smectite pulps. Optimum conditions for improved, orthokinetic flocculation performance of negatively charged clay particles, reflecting faster settling flocs comprised (i) coagulation, (ii) moderate agitation rate, (iii) shorter agitation time, and (iv) anionic rather than non-ionic PAM. The optimum dewatering rates were significantly higher than those produced by standard, manual-mixing flocculation techniques (plunging and cylinder inversion) commonly used in industry for flocculant trials. The optimum flocculation conditions did not, however, have a significant impact on the final sediment solid content of 20-22 wt%. Further application of shear to pre-sedimented pulps improved consolidation by 5-7 wt% solid. Higher shear yield stresses and greater settling rates were displayed by PAM A based than PAM N based pulps and this is attributed to the former's more expanded interfacial conformation and greater clay particles bridging ability. It appears that the intrinsic clay particles' physico-chemical properties and interactions limit compact pulp consolidation. PMID- 16038922 TI - Interfacial tension in phase-separated aqueous cationic/anionic surfactant mixtures. AB - Interfacial tensions in two aqueous phase-separated cationic/anionic surfactant mixtures, CTAB/AS and 12-3-12/AS, without and with NaBr added were determined by the spinning drop method at 318.15 K. CTAB, 12-3-12 and AS are the abbreviations for cetyltrimethylammonium bromide, 1,3-propanediyl-bis(dodecyldimethylammonium bromide) and sodium dodecyl sulfonate, respectively. The interfacial tension sigma was found to be in the range of 0.06-21 microNm(-1). Toward a better understanding of the influence of the concentration difference between the separated phases in aqueous two-phase systems (ATPS) to interfacial tension, compositions of equilibrium phases were determined by elemental analysis coupled with material balance and electroneutrality. The investigation indicates that the concentration differences of surfactant ions between the separated phases and the adsorption of surfactant ions at the interface are the decisive factors determining the magnitude of interfacial tension. PMID- 16038923 TI - Oxidation behaviors of metallic copper particles in NO reduction mechanism of copper/activated carbons. AB - The reduction of nitric oxide (NO) over Cu/ACFs, prepared by copper electroplating, has been studied. It is found that copper content on the ACFs increases with increasing plating time (up to 45 wt%), while the textural properties including specific surface areas and total pore volumes decrease. As an experimental result, the NO reduction efficiency is increased in all of the Cu/ACFs, and it is confirmed that NO is converted into nitrogen and oxygen on the Cu/ACF surfaces (at 500 degrees C). Especially, the Cu metals on the ACF surfaces scavenge oxygen by oxidizing themselves into Cu2O and finally CuO as a reductant. It is indicated that copper metals on the Cu/ACFs play a major role in the NO removal in this system. PMID- 16038924 TI - Contribution of micelles to ionic strength of surfactant solution. AB - A new definition of the concentration-dependent "shielding factor" has been proposed, along with its particular value at a concentration assumed to be equal to critical micelle concentration (cmc). It has been shown that herein defined shielding factor enables one to calculate the values of monomer (a(x)) and counterion activities (a(y)) which are in better agreement with the measured ones than those calculated using Burchfield and Wooley's constant value of the shielding factor. In addition, it was shown that by using particular value of shielding factor at cmc, one is able to calculate the micellar charge radius which is dependent on aggregation number, counterion parameter and cmc, all of them having important implications for thermodynamic treatment of the micellization process. PMID- 16038925 TI - The wettability of polytetrafluoroethylene and polymethyl methacrylate by aqueous solution of two cationic surfactants mixture. AB - Advancing contact angle (theta) measurements were carried out for aqueous solutions of cetyltrimethylammonium bromide (CTAB) and cetylpyridinium bromide (CPyB) mixtures on polytetrafluoroethylene (PTFE) and polymethyl methacrylate (PMMA). The obtained results indicate that the wettability of PTFE and PMMA by aqueous solutions of CTAB and CPyB mixtures depends on the composition and concentration of the mixture; however, synergism in the wettability does not exist. In the range of low concentrations of aqueous solution mixtures there is a linear dependence between the contact angle and composition of the mixtures, but at a concentration close to CMC a deviation from linear dependence is observed. In contrast to Zisman, there is no linear dependence between costheta and the surface tension of aqueous solution of CTAB and CPyB mixtures, but a linear dependence exists between the adhesional and surface tension, and these lines have a slope -1 and -0.34 for PTFE and PMMA, respectively, which suggests that adsorption of CTAB and CPyB mixtures at water-air and PTFE-water is the same, and the orientation of the CTAB and CPyB molecules at both interfaces in the saturated monolayer should also be the same. Adsorption of these mixtures at water-air interface is considerably higher than at PMMA-water interface, and CTAB and CPyB molecules should be parallelly oriented to PMMA surface in the saturated monolayer. Extrapolation of the straight lines to the points corresponding to the surface tension of aqueous solution, which completely spreads over the PTFE and PMMA surface, gives a critical surface tension of wetting equal to 23.44 and 33.13 mN/m, respectively. The value of 23.44 mN/m is higher than that of the surface tension of PTFE, but the value of 33.13 is lower than that of Lifshitz van der Waals components of PMMA surface tension. On the basis of the critical surface tension, the surface tension of PTFE and PMMA, the Young equation, and thermodynamic analysis of the adhesion work of aqueous solution of surfactant to polymer surface, it was found that for PTFE and PMMA the changes of the contact angle of aqueous solution of two cationic surfactants mixtures on their surfaces as a function of the solution concentration resulted only from the decrease of the polar component of the solution surface tension. PMID- 16038926 TI - Culture and comparison of feline myeloid dendritic cells vs macrophages. AB - To gain insight into the role of dendritic cells (DCs) in feline immunodeficiency virus (FIV) infection and immunity, methods were developed to culture feline myeloid DCs from CD14(+) monocytes with a combination of human recombinant granulocyte-macrophage colony-stimulating factor (hrGM-CSF) and interleukin-4 (hrIL-4). These cells were compared with feline macrophages cultured in the presence of hrGM-CSF. As with DCs in other species, feline DCs showed uniformly high MHC class II expression, moderate B7.1 expression, potent induction of the allogeneic mixed leucocyte reaction (MLR), and moderate uptake of fluorescein isothiocyanate-dextran (FITC-DX) in the endocytic assay. In comparison with feline macrophages, DCs showed higher expression of MHC class II, similar expression of B7.1, CD14, CXCR4 and CD1a, and lower expression of CD11b. When placed on alcian blue-coated glass slides, DCs differed from macrophages in showing a greater tendency to spread out; they also had characteristic fine cytoplasmic processes instead of the broader pseudopodia of macrophages. Basal IL 12 mRNA expression and FITC-DX uptake were greater in DCs than in macrophages. Unlike feline DCs, feline macrophages exhibited a dose-dependent suppressive effect in the MLR. Feline DCs propagated in vitro should prove useful in the development of DC-mediated vaccination and therapy for infectious and neoplastic feline diseases. Additionally, macrophages cultured with GM-CSF provide a potential means of studying the mechanism of immunosuppression in cats. PMID- 16038927 TI - Phoneme awareness is a key component of alphabetic literacy skills in consistent and inconsistent orthographies: evidence from Czech and English children. AB - Two studies investigated the importance of phoneme awareness relative to other predictors in the development of reading and spelling among children learning a consistent orthography (Czech) and an inconsistent orthography (English). In Study 1, structural equation models revealed that Czech (n=107) and English (n=71) data were fitted well by the same predictors of reading and spelling. Phoneme awareness was a unique predictor in all models. In Study 2, Czech (n=40) and English (n=27) children with dyslexia showed similar deficits on phoneme awareness relative to their age- and spelling-matched control peers. Phoneme awareness appears to be a core component skill of alphabetic literacy, which is equally important for learners of consistent and inconsistent orthographies. PMID- 16038928 TI - Monoclonal antibody 5C3 raised against formalin fixed paraffin-embedded invasive breast tumour tissue: characterisation of its reactive antigen via immunoprecipitation and internal sequencing. AB - Monoclonal antibodies (MAbs) provide a powerful tool for the identification of novel tumour associated antigens. In an attempt to identify such an antigen, MAbs were generated by immunization with paraffin wax-embedded formalin-fixed invasive ductal breast tumour tissue from a patient who relapsed following an initial response to adjuvant chemotherapy. Extensive immunocytochemical and Western blot analysis of a range of cell lines and tissues including a series of pre- and post chemotherapy treated invasive ductal breast carcinomas, with one of these MAbs, antibody 5C3, indicated that the 5C3 reactive antigen displayed a wide spectrum of reactivity amongst various human tumours. A reduced level of 5C3 expression was observed in non-cancerous archival breast tissues and breast cell lines and normal murine tissues compared to the expression observed in infiltrating breast tumour cells. Immunoprecipitation studies using the human ductal breast carcinoma cell line, ZR-75-1 resulted in the isolation of a 175 kDa reactive band which was excised from an SDS-PAGE gel and subjected to internal sequencing. Sequencing analysis and database searching revealed that this 175 kDa band represented a cytokeratin heteropolymer, composed of type I cytokeratin 9 and type II cytokeratin 6. Further studies confirmed that antibody 5C3 recognised this heteropolymer of cytokeratin 9 and 6 but not the individual cytokeratins. This novel method of MAb generation may facilitate the isolation of further potentially interesting cellular antigens. Characterisation of these novel antigens may identify specific disease targets with possible prognostic or predictive significance. PMID- 16038929 TI - An improved methodology to detect human T cell receptor beta variable family gene expression patterns. AB - Comprehensive gene expression analysis of the T cell receptor repertoire of an individual can be very useful in evaluating the immune response in a variety of conditions. Antibody-based analysis methods can detect approximately 60% of the human T cell receptor beta variable (TCRBV) proteins, while gene expression analysis, primarily through employment of the polymerase chain reaction (PCR), has had somewhat greater success in the detection of additional TCRBV families. Many of these previous PCR methods, however, have been unable to detect all 91 alleles of the human TCRBV genes. This is primarily due to either deficiencies in the amplification of all of the variable beta families, subfamilies, and alleles, or the prior lack of a systematic classification of the TCR variable family gene segment sequences. We describe here a real-time reverse transcription polymerase chain reaction-based method, which allows efficient automation and integration of amplification, detection, and analysis with sequence-specific detection of all T cell receptor beta variable gene families, subfamilies, and alleles. This method, which in itself contributes significant improvements over existing technologies through its comprehensiveness and efficiency, also functions independently of variables such as sample source and sample processing and has the ability to run on multiple real-time PCR platforms, affording one the implementation of personal preferences. PMID- 16038930 TI - Crystal structure of a putative type I restriction-modification S subunit from Mycoplasma genitalium. AB - The crystal structure of the eubacteria Mycoplasma genitalium ORF MG438 polypeptide, determined by multiple anomalous dispersion and refined at 2.3 A resolution, reveals the organization of S subunits from the Type I restriction and modification system. The structure consists of two globular domains, with about 150 residues each, separated by a pair of 40 residue long antiparallel alpha-helices. The globular domains correspond to the variable target recognition domains (TRDs), as previously defined for S subunits on sequence analysis, while the two helices correspond to the central (CR1) and C-terminal (CR2) conserved regions, respectively. The structure of the MG438 subunit presents an overall cyclic topology with an intramolecular 2-fold axis that superimposes the N and the C-half parts, each half containing a globular domain and a conserved helix. TRDs are found to be structurally related with the small domain of the Type II N6 adenine DNA MTase TaqI. These relationships together with the structural peculiarities of MG438, in particular the presence of the intramolecular quasi symmetry, allow the proposal of a model for S subunits recognition of their DNA targets in agreement with previous experimental results. In the crystal, two subunits of MG438 related by a crystallographic 2-fold axis present a large contact area mainly involving the symmetric interactions of a cluster of exposed hydrophobic residues. Comparison with the recently reported structure of an S subunit from the archaea Methanococcus jannaschii highlights the structural features preserved despite a sequence identity below 20%, but also reveals important differences in the globular domains and in their disposition with respect to the conserved regions. PMID- 16038931 TI - Assembly of motor proteins, PomA and PomB, in the Na+-driven stator of the flagellar motor. AB - PomA and PomB are transmembrane proteins that form the stator complex in the sodium-driven flagellar motor of Vibrio alginolyticus and are believed to surround the rotor part of the flagellar motor. We constructed and observed green fluorescent protein (GFP) fusions of the stator proteins PomA and PomB in living cells to clarify how stator proteins are assembled and installed into the flagellar motor. We were able to demonstrate that GFP-PomA and GFP-PomB localized to a cell pole dependent on the presence of the polar flagellum. Localization of the GFP-fused stator proteins required their partner subunit, PomA or PomB, and the C-terminal domain of PomB, which has a peptidoglycan-binding motif. Each of the GFP-fused stator proteins was co-isolated with its partner subunit from detergent-solubilized membrane. From these lines of evidence, we have demonstrated that the stator proteins are incorporated into the flagellar motor as a PomA/PomB complex and are fixed to the cell wall via the C-terminal domain of PomB. PMID- 16038932 TI - Crystal structure of an Anti-meningococcal subtype P1.4 PorA antibody provides basis for peptide-vaccine design. AB - In various western countries, subtype P1.4 of Neisseria meningitidis serogroup B causes the greatest incidence of meningococcal disease. To investigate the molecular recognition of this subtype, we crystallised a peptide (P1HVVVNNKVATH(P11)), corresponding to the subtype P1.4 epitope sequence of outer membrane protein PorA, in complex with a Fab fragment of the bactericidal antibody MN20B9.34 directed against this epitope. Structure determination at 1.95 A resolution revealed a unique complex of one P1.4 antigen peptide bound to two identical Fab fragments. One Fab recognises the putative epitope residues in a 2:2 type I beta-turn at residues P5NNKV(P8), whereas the other Fab binds the C terminal residues of the peptide that we consider a crystallisation artefact. Interestingly, recognition of the P1.4 epitope peptide is mediated almost exclusively through the complementarity-determining regions of the heavy chain. We exploited the observed turn conformation for designing conformationally restricted cyclic peptides for use as a peptide vaccine. The conformational stability of the two peptide designs was assessed by molecular dynamics simulations. Unlike the linear peptide, both cyclic peptides, conjugated to tetanus toxoid as a carrier protein, elicited antibody responses in mice that recognised meningococci of subtype P1.7-2,4. Serum bactericidal assays showed that some, but not all, of the sera induced with the cyclic peptide conjugates could activate the complement system with titres that were very high compared to the titres induced by complete PorA protein in its native conformation administered in outer membrane vesicles. PMID- 16038933 TI - Phenotypic variation between parent-offspring trios and non-trios in genetic studies of schizophrenia. AB - BACKGROUND: Phenotypic differences between parent-offspring trios and non-trios have been reported for various psychiatric disorders, and it has been suggested that this may make comparisons of case-control and family-based results for gene disease association studies inappropriate. AIMS: To compare phenotypes between trios and non-trios with schizophrenia, and explore possible reasons for differences observed. METHOD: Phenotypes were compared between trios (n=75) and non-trios (n=424) collected as part of a case-control study. RESULTS: Differences were observed for most phenotypes investigated, although all were eliminated after adjusting for confounding. CONCLUSIONS: Confounding, genetic heterogeneity or selection bias could result in differences in case-control and family-based results. However as we discuss, where adequately designed case-control studies are used, gene-disease association results would be incomparable between family based and case-control studies only if genetic heterogeneity was present. These results do not support the presence of such genetic heterogeneity in schizophrenia. PMID- 16038934 TI - The development of an instrument measuring unintentional injuries in young children in low-income settings to serve as an evaluation tool for a childhood home injury prevention program. AB - PROBLEM: Responding to the lack of standardized instrumentation, especially as applied in low-income contexts, the researchers decided that an instrument should be developed to serve as an evaluation tool for a childhood (unintentional) injury prevention program as well as a tool from which injury risks (poisonings, burns and falls) could be identified within households. METHODS: Cross-sectional samples of 521 households in four low-income sites in Cape Town and Johannesburg, South Africa, participated in the three phases of the study. The generation of an item pool based on a table of specifications, subsequent scientific item reduction procedures, reviews from experts and pilot tests were used to develop the scales for measuring the injury risks. RESULTS: The developed instrument complies with all the requirements for a valid and reliable measurement instrument. CONCLUSION: The instrument allows valid comparison of risks between communities, as well as before and after comparisons for an intervention program. IMPACT: The provision of this instrument may bring enormous benefits to research studies. It can also provide proactive, rather than reactive, information about injury risks before they develop into injuries and thus allows focusing of safety efforts for improvement of problematic areas in the households. PMID- 16038935 TI - Synthesis and recommendations of the economic evaluation of OHS interventions at the company level conference. AB - PROBLEM: In today's economic environment, enterprises may not be able to fund every new project aimed at promoting health and safety in the workplace. Company level economic evaluation of interventions can provide guidance in sound business decision-making. The Economic Evaluation of Occupational Health and Safety Interventions at the Company Level Meeting brought together members of the global occupational safety and health community interested in encouraging the use of economic knowledge and tools to evaluate economic gains from occupational health and safety interventions. DISCUSSION: Discussions of the six models presented explored similarities, reliability, and potential use by corporate enterprises, small and medium enterprises, developing and transitioning nations, and economic theorists. Each group provided specific projects that could be pursued to advance knowledge in the area of economic evaluation at the company level. CONCLUSION: This conference established pathway to incorporate economic evaluation of health and safety interventions or programs at the workplace. PMID- 16038936 TI - Understanding the return on health, safety and environmental investments. AB - INTRODUCTION: Traditionally, health safety and environmental (HS & E) investments have been viewed as expensive but necessary. HS & E professionals had a difficult task of providing data showing that these investments can also contribute to business success. An ORC task force developed a way for traditional financial analysis methods to be applied to HS & E investments and decisions. METHOD: The result of the task force effort is the software called the ORC Return on Health, Safety and Environmental Investments (ROHSEI), a supporting tool to provide a comprehensive look at health, safety or environmental investment projects and their potential financial impacts. RESULTS: Since its development, more than 200 companies, government agencies, and educational institutions have been trained in the ROHSEI process and software. CONCLUSIONS: HS & E professionals who are able to evaluate and communicate the business value of health, safety and environmental efforts will improve understanding of the impacts HS & E projects and contribute to better decision making by their organizations. PMID- 16038937 TI - Net-cost model for workplace interventions. AB - PROBLEM: Few methods exist for comprehensively examining the costs and benefits of ergonomic interventions applicable to a variety of economic sectors and settings. METHODS: An instrument for data collection and data analysis at the facility level is presented. In this net-cost model intervention costs are defined by equipment and labor costs for the interventions as well as the avoided costs of lost work time, medical care, and productivity improvements. RESULTS: Net-cost estimates for three case studies show that ergonomic interventions applied appropriately can result in substantial cost savings for the companies. DISCUSSION: It would be prudent to incorporate a protocol for collecting cost and effectiveness data in the standard operating procedures of companies introducing ergonomic interventions. Validation of the net-cost model through prospective studies is necessary. IMPACT ON INDUSTRY: This model may be used to determine the net-cost of implemented or proposed ergonomic interventions in industrial facilities. PMID- 16038938 TI - An overview to CERSSO's self evaluation of the cost-benefit on the investment in occupational safety and health in the textile factories: "a step by step methodology". AB - INTRODUCTION: The Pan American Health Organization (PAHO) and CERSSO collaborated to develop a new Tool Kit (TK), which became available in May 2002. PAHO already had a TK in place, and CERSSO requested that one be developed for their needs. CERSSO wanted to enable managers and line workers in garment factories to self diagnose plant and workstation hazards and to estimate the costs and benefits of investing in occupational safety and health (OSH) as a way to improve productivity and competitiveness. METHODS: For consistency, the collaborating organizations agreed to construct the TK according to PAHO's methodology. The instrument was developed to be comprehensive enough that any user can collect the data easily. It integrates epidemiologic, risk assessment, clinic, engineering, and accountability issues, organized to include step-by-step training in: (a) performing risk assessments in the workplaces (risk factors); (b) making cause effect relationships; (c) improving decision making on OSH interventions; (d) doing calculations of direct and indirect costs and savings; and (e) doing calculation of the overall cost-benefit of OSH interventions. RESULTS AND CONCLUSIONS: Since July 2002, about 2,400 employees and officials from 736 garment factories, Ministries of Labor, Health, Social Security Institutes, and Technical Training Institutions of Central America and the Dominican Republic have used this instrument. Systematically, they have calculated a positive relationship of the investment (3 to 33 times). Employers are now aware of the financial rewards of investing in OSH. The TK is available in Spanish, Korean, and English. In July 2003, a software program in Spanish and English was developed (180 persons have been trained in the region), which requires less time to execute with better reliability. PMID- 16038939 TI - The productivity assessment tool: computer-based cost benefit analysis model for the economic assessment of occupational health and safety interventions in the workplace. AB - INTRODUCTION AND METHOD: This paper describes the concepts behind cost benefit analysis in occupational health and safety and introduces the Productivity Assessment Tool, a method by which an analysis may be performed relatively easily in a service or manufacturing workplace. The advantage of using such analyses is to show the important financial role that safe and efficient workplaces play in the workplace. RESULTS: By using analytical tools, the effectiveness of an intervention (workplace change) may be estimated prior to its introduction. IMPACT ON INDUSTRY: This places occupational health and safety on the same financial footing as other proposed workplace changes and thus places occupational health and safety in a strong position to attract scarce resources. PMID- 16038940 TI - The behavior of the human erythrocyte as an imperfect osmometer: a hypothesis. AB - The human erythrocyte does not behave as a perfect osmometer that is its volume does not change as predicted with the change of the tonicity of the medium, as if there was a fraction of the cell water not participating in the osmotic exchange. A mechanism of control of the erythrocyte shape has been previously proposed in which Band 3 (AE1), the protein anion exchanger of Cl(-) and HCO(3)(-), plays a central role. Specifically, decrease and increase of the ratio of its outward facing conformation and inward-facing conformation (Band 3(o)/Band 3(i)) contract and relax the membrane skeleton, thus favoring echinocytosis and stomatocytosis, respectively. The equilibrium Band 3(o)/Band 3(i) ratio is determined by the Donnan equilibrium ratio of anions and protons, increasing with it (r=Cl(i)( )/Cl(o)(-)=HCO 3(i)(-)/HCO 3(o)(-)=H(o)(+)/H(i)(+)). The Donnan ratio is influenced by the erythrocyte transport and metabolic activities. The volume change of the human erythrocyte alters the skeleton conformation as it is accompanied by a change of the membrane curvature. Thus, the mechanism could be a hypothesis for explaining the behavior of the human erythrocyte as an imperfect osmometer since the Donnan ratio controls the Band 3(o)/Band 3(i) ratio which controls the volume by a control of the degree of contraction or relaxation of the skeleton. Predictions made by the hypothesis on the Ponder's coefficient R' values in the presence of sucrose or Band 3 substrates slowly transported as well as on the participation of Band 3 in the osmotic hemolysis appear to be corroborated by previous observations. If the hypothesis was valid, it would follow that there is a pressure gradient across the erythrocyte membrane. The equilibrium volume is antagonistically determined by the Donnan ratio per se and Band 3. Band 3, rather than the ratio of surface-to-volume, primarily controls the osmotic hemolysis. PMID- 16038941 TI - Simplified models for gas exchange in the human lungs. AB - This paper presents a hierarchy of models with increasing complexity for gas exchange in the human lungs. The models span from a single compartment, inflexible lung to a single compartment, flexible lung with pulmonary gas exchange. It is shown how the models are related to well-known models in the literature. A long-term purpose of this work is to study nonlinear phenomena seen in the cardio-respiratory system (for example, synchronization between ventilation rate and heart rate, and Cheyne-Stokes respiration). The models developed in this paper can be regarded as the controlled system (plant) and provide a mathematical framework to link between "molecular-level", and "systems level" models. It is shown how changes in molecular level affect the alveolar partial pressure. Two assumptions that have previously been made are re-examined: (1) the hidden assumption that the air flow through the mouth is equal to the rate of volume change in the lungs, and, (2) the assumption that the process of oxygen binding to hemoglobin is near equilibrium. Conditions under which these assumptions are valid are studied. All the parameters in the models, except two, are physiologically realistic. Numerical results are consistent with published experimental observations. PMID- 16038942 TI - Extract from Clerodendron colebrookianum Walp protects rat heart against oxidative stress induced by ischemic-reperfusion injury (IRI). AB - Reactive oxygen species (ROS) have pathogenic effects on ischemic-reperfusion injury of heart. Hence, it is important to identify natural antioxidative agents to mitigate such effects. Recently, it has been reported that Clerodendron colebrookianum (CC) leaf extract has antioxidant and hypolipidemic effects in experimental animals. The aim of this study was to examine whether acute treatment with CC extract offers protection against ischemic-reperfusion injury (IRI) and IRI-induced changes in endogenous antioxidant enzyme activities of rat heart. Isolated rat hearts were perfused using the Langendorff's technique, and 20 min of global ischemia was followed by 40 min of reperfusion. Lipid peroxidation after the ischemic-reperfusion episode was significantly reduced in the CC extract-treated heart compared to the control group and suppressed the leakage of lactate dehydrogenase (LDH) during reperfusion. Moreover, CC extract diminished the depletion of myocardial antioxidant enzymes (SOD, Catalase, GSH and GPx) after ischemia-reperfusion. Furthermore, IRI-induced cellular damage was significantly less in CC extract treated myocytes. These results indicate that CC leaf extract protects against oxidative stress and cellular injury associated with ischemic-reperfusion injury of rat heart and suggests that the protective effects of CC extract depend on its antioxidant properties. PMID- 16038943 TI - Emerging chemicals of concern: pharmaceuticals and personal care products (PPCPs) in Asia, with particular reference to Southern China. AB - In many western nations, pharmaceuticals and personal care products (PPCPs) are present in aquatic environments, raising concerns amongst chemists and toxicologists regarding their potential environmental fates and effects. However, there are few published reports of PPCPs in environmental samples from Southeast Asia. Whilst the environmental toxicology of PPCPs is not well understood, several effects cause concern, such as feminisation or masculinisation by hormones and xenoestrogens, synergistic toxicity from complex mixtures at low concentrations, potential creation of resistant strains in natural bacterial populations, and other potential concerns for human health. Whilst both the presence and distributions of PPCPs in Southeast Asia and China are not well known, observations elsewhere suggest that they may be important contaminants in the aquatic environment. This is particularly emphasised by the enormous production and widespread use of many PPCPs in China, particularly antibiotics utilised in human and veterinary medicine applications. This Viewpoint presents a general description of the issue, characterises the current status of PPCP analyses and reporting in the Southeast Asian region, and proposes a recommended approach for monitoring and chemical assessment of one group of PPCPs, antibiotics, in the aquatic environments of Hong Kong and the Pearl River Delta. PMID- 16038944 TI - Seasonal variability of benthic indices: an approach to test the applicability of different indices for ecosystem quality assessment. AB - In recent studies, several benthic biological indices were developed or used to assess the ecological quality status of marine environments. In the present study the seasonal variability of several univariate and multimetric indices was studied on a monthly scale (September 2000 until May 2002) in different areas of the North Sea such as the German Bight, the Oyster Ground and the Dogger Bank. The stations were chosen to reflect a gradient in the hydrographic regime, temperature and organic matter supply. The seasonal variability was highest for the univariate indices such as the Shannon-Wiener Index and the Hurlbert Index. Thus, due to sensitivity to recruitment the corresponding ecological status ranged from 'good' to 'poor' depending on the season. For the multimetric indices such as the AMBI or the BQI the seasonal variability and the corresponding ecological status were low. The results are discussed concerning possible consequences for ecological quality assessment especially related to the Water Framework Directive (WFD). PMID- 16038946 TI - Requirement of water-flow for sustainable growth of Pocilloporid corals during high temperature periods. AB - Reef-building corals are threatened worldwide by mass-scale coral bleaching episodes that are pronounced in high sea surface temperature (SST) conditions. Although water-flow has been suggested to be a mitigating factor for bleaching, long-term effects of flow-mediated bleaching suppression are as yet not fully understood. In order to investigate flow effects, we monitored the corals Pocillopora damicornis and Stylophora pistillata grown for 20 months in experimental outdoor flumes with the flow rates of 20 cms(-1) (flow) and <3 cms( 1) (still). Although bleaching was observed under high SST conditions, both species showed a shorter period or entirely no visible bleaching under the flow conditions. Better colony growth was found in the flow conditions whereas significant growth suppression and higher mortality were observed in still conditions. We conclude that water-flow is an essential environmental factor for the corals P. damicornis and S. pistillata, especially under high SST conditions. PMID- 16038945 TI - Effect of an acid mine drainage effluent on phytoplankton biomass and primary production at Britannia Beach, Howe Sound, British Columbia. AB - We investigated the effect of acid mine drainage (AMD) from an abandoned copper mine at Britannia Beach (Howe Sound, BC, Canada) on primary productivity and chlorophyll a levels in the receiving waters of Howe Sound before, during, and after freshet from the Squamish River. Elevated concentrations of copper (integrated average through the water column >0.050 mgl(-1)) in nearshore waters indicated that under some conditions a small gyre near the mouth of Britannia Creek may have retained the AMD from Britannia Creek and from a 30-m deep water outfall close to shore. Regression and correlation analyses indicated that copper negatively affected primary productivity during April (pre-freshet) and November (post-freshet). Negative effects of copper on primary productivity were not supported statistically for July (freshet), possibly because of additional effects such as turbidity from the Squamish River. Depth-integrated average and surface chlorophyll a were correlated to copper concentrations in April. During this short study we demonstrated that copper concentrations from the AMD discharge can negatively affect both primary productivity and the standing stock of primary producers in Howe Sound. PMID- 16038947 TI - Assessing reference conditions and physico-chemical status according to the European Water Framework Directive: a case-study from the Basque Country (Northern Spain). AB - The European Water Framework Directive requires the development of new and accurate methodologies, addressing the assessment of the physico-chemical status of transitional and coastal waters; these are considered by the Directive as the supporting elements for the final evaluation of the Ecological Quality Status. This contribution develops new approaches in the determination of the physico chemical status, solving some problems detected in previous contributions, i.e.: (a) fitting the classification of water bodies and typologies, by means of the stretching of the typologies, according to the natural salinity gradient of types; (b) defining reference conditions, based upon the new approach to typologies, (c) proposing accurate multivariate methodologies, in determining the physico-chemical status of the transitional and coastal waters, based upon the defined typologies and references; and (d) discussion of the results obtained by reference to methodological aspects and water quality evolution in the Basque Country, Spain (as a case-study), during the last decade. PMID- 16038948 TI - Aliphatic and aromatic hydrocarbons in marine biota and coastal sediments from the Gulf and the Gulf of Oman. AB - The composition and spatial distribution of aliphatic and polycyclic aromatic hydrocarbons (PAHs) were investigated in biota and coastal sediments from four countries surrounding the Gulf (Bahrain, Qatar, United Arab Emirates and Oman). The levels of total petroleum hydrocarbons (TPH), aliphatic unresolved mixture and PAHs in sediments and biota were relatively low compared to world-wide locations reported to be chronically contaminated by oil. Only in the case of the sediments collected near the BAPCO oil refinery in Bahrain, having concentrations of 779 microg g(-1) total petroleum hydrocarbon equivalents and 6.6 microg g(-1) Sigma PAHs, can they be categorized as chronically contaminated. Some evidence of oil contamination was also apparent in sediments and bivalves around Akkah Head and Abu Dhabi in the UAE, and near Mirbat in Oman. Contaminant patterns in sediments and biota indicated that the PAHs were mainly from fossil sources, with the exception of the high PAH concentrations in sediments near the BAPCO refinery that contained substantial concentrations of carcinogenic PAH combustion products. PMID- 16038949 TI - Multi-scale modeling of the follicle selection process in the ovary. AB - The biological meaning of follicular development is to free fertilizable oocytes at the time of ovulation. The ovulation rate results from an FSH-dependent follicle selection process. In this paper, we designed a multi-scale model of follicular development, where selection arises from the endocrine feedback between the ovaries and pituitary gland and appeals to control theory concepts. Each ovarian follicle is described through a 2D density function giving an age and maturity-structured description of its cell population. The control intervenes in the velocity, gain and loss terms of the conservation law ruling the changes in the density. The model accounts for the changes in the total cell number, growth fraction and global maturity of both ovulatory and degenerating follicles for various intensities of the selection rate. The different selection process outputs (mono- or poly-ovulation, anovulation) predicted by the model are consistent with physiological knowledge regarding vascularization, pituitary sensitivity to ovarian feedback and treatment with exogenous FSH. PMID- 16038950 TI - Processing of auditory spatial cues in human cortex: an fMRI study. AB - The issue of where in the human cortex coding of sound location is represented still is a matter of debate. It is unclear whether there are cortical areas that are specifically activated depending on the location of sound. Are identical or distinct cortical areas in one hemisphere involved in processing of sounds from the left and right? Also, the possibility has not been investigated so far that distinct areas have a preference for processing of central and eccentric sound locations. The present study focussed on these issues by using functional magnetic resonance imaging (fMRI). Activations evoked by left, right and central sounds were analysed separately, and contrasts were computed between these conditions. We did not find areas, which were involved in the processing of exclusively left, right or central sound positions. Large overlapping areas rather were observed for the three sound stimuli, located in the temporal, parietal and frontal cortices of both hemispheres. This result argues for the idea of a widely distributed bilateral network accessing an internal representation of the body to encode stimulus position in relation to the body median plane. However, two areas (right BA 40 and left BA 37) also were found to have preferences for sound position. In particular, BA 40 turned out to be significantly more activated by processing central positions, compared to eccentric stimuli. In line with previous findings on visual perception, the latter observation supports the assumption that the right inferior parietal cortex may be preferentially involved in the perception of central stimulus positions in relation to the body. PMID- 16038951 TI - In rats, sighs correlate with relief. AB - A deep breath, i.e., a sigh, in mammals is a ubiquitous respiratory phenomenon, whose function is to prevent airlessness (atelectasis) in hypoventilated parts of lungs. Sighs are also correlated with emotions, such as anxiety, anger and resentment in humans and obviously, judging from the expression--sigh of relief- in many languages, with relaxation or relief. If sighs are indiscriminately associated with opposite emotions, their role in social communication is doubtful. If, however, there is a selective facilitation of sighs by either fear, anxiety or relief, then they might, in addition to their respiratory role, function also as a social signal of a particular mood. To induce fear a stimulus was paired with a tail shock (5 times in a daily session). To provide a relief, another stimulus, presented before the expected shock (also 5 times per session), was followed by the omission of shock. In 16 rats experiencing fear during a Danger Stimulus (predictor of tail shock) and a relief during the Safety Stimulus (predictor of the non-occurrence of expected shock) the rate of sighing was 7.5 times higher during relief (180/h) than during fear (24/h), and 20 times higher than between trials (9/h), with all differences highly significant (p<0.001). This clear correlation of sighs with relief (from fear of the tail shock) supports our hypothesis that sighs in social mammals may function as signals of safety. PMID- 16038952 TI - Polar acyclic diterpenoids from Bifurcaria bifurcata (Fucales, Phaeophyta). AB - From the lipophilic extract of the brown alga Bifurcaria bifurcata collected off the Atlantic coast of Southern Brittany (Quiberon, France), five polar linear diterpenoids have been isolated. These metabolites have been identified as hydroxylated derivatives of 13-oxo- and 13-hydroxygeranylgeraniol. Their structures were characterized on the basis of chemical and spectral evidence including two-dimensional NMR experiments and mass spectrometric techniques. The absolute configuration of the 13-position has been determined, for the 13 hydroxygeranylgeraniol derivatives, to be R by means of a modified Mosher's method and therefore that of 13-hydroxygeranylgeraniol (eleganediol) has been revised. Along with these compounds, three related known geranylgeraniol derivatives were also identified, and these data were used for chemotaxonomical purposes. PMID- 16038953 TI - Suppression of Wolffia arrhiza growth by brassinazole, an inhibitor of brassinosteroid biosynthesis and its restoration by endogenous 24 epibrassinolide. AB - The effect of the brassinosteroid (BR) 24-epibrassinolide (epiBL; 10(-13)-10( 6)M) on growth and levels of chlorophylls, carotenoids, sugars and protein in Wolffia arrhiza after 7 days of cultivation is reported. Application of epiBL to W. arrhiza cultures stimulates the growth and increases the content of photosynthetic pigments, sugar and protein. The greatest effect of epiBL is observed at a concentration of 10(-9)M. We tested the action of Brz2001, a specific BR biosynthesis inhibitor, in the range of 10(-6)-10(-4)M. Addition of Brz2001 to W. arrhiza cultures inhibits their growth after 7 days of cultivation. The inhibition of growth could be reversed by the addition of epiBL. Moreover, there was not complete recovery to the level of control, especially at 5 x 10(-5) 10(-4)M Brz2001. The effects of treatment with 10(-9)M epiBL mixed with a mevalonate pathway inhibitor (mevinolin), or a 2-methylerythritol 4-phosphate pathway inhibitor (clomazone), were also investigated. Mevinolin did not inhibit growth of W. arrhiza after 7 days of cultivation. However, clomazone did. Addition of epiBL overcame this inhibition. These results suggest that the mevalonate pathway may not function well in W. arrhiza and that biosynthesis of BRs through the non-mevalonate pathway in W. arrhiza could be possible. PMID- 16038954 TI - Benzopyrans from Curvularia sp., an endophytic fungus associated with Ocotea corymbosa (Lauraceae). AB - An isolate of Curvularia sp. was obtained from the leaves of Ocotea corymbosa, a native plant of the Brazilian Cerrado. The ethyl acetate extract from culture of this fungus afforded two benzopyran derivatives: (2'S)-2-(propan-2'-ol)-5-hydroxy benzopyran-4-one (2) and 2,3-dihydro-2-methyl-benzopyran-4,5-diol (4); and two known benzopyrans: 2-methyl-5-methoxy-benzopyran-4-one (1) and (2R)-2,3-dihydro-2 methyl-5-methoxy-benzopyran-4-one (3). The structures of 2 and 4 were established on the basis of comprehensive spectroscopic analysis, mainly using 1D and 2D NMR experiments. The benzopyrans 1 and 2 showed weak in vitro antifungal activity against Cladosporium sphaerospermum and C. cladosporioides. Analyses of the biological activities were also carried out on HeLa (human cervix tumor) and CHO (Chinese hamster ovary) cells, aiming to evaluate their potential effects on mammalian cell line proliferation. Results from both cell lines indicated that compound 2 was able to induce cell proliferation: 70% on HeLa cells and 25% on CHO cells. PMID- 16038955 TI - Identification and quantitative analysis of beta-sitosterol oxides in vegetable oils by capillary gas chromatography-mass spectrometry. AB - As vegetable oils and phytosterol-enriched spreads are marketed for frying food or cooking purposes, temperature is one of the most important factors leading to the formation of phytosterol oxides in food matrix. A methodology based on saponification, organic solvent extraction, solid-phase extraction (SPE), followed by mass spectrometric identification and quantitation of beta-sitosterol oxides using capillary gas chromatography-mass spectrometry (GC-MS) in selected ion monitoring (SIM) mode was developed and characterized. Relative response factors of six beta-sitosterol oxides, including 7alpha-hydroxy, 7beta-hydroxy, 5,6alpha-epoxy, 5,6beta-epoxy, 7-keto, and 5alpha,6beta-dihydroxysitosterol, were calculated against authentic standards of 19-hydroxycholesterol or cholestanol. Linear calibration data, limit of detection, and sample recoveries during analytical process. Recoveries of these oxidation compounds in spiked samples ranged from 88 to 115%, while relative standard derivation (R.S.D.) values were below 10% in most cases. The analytical method was applied to quantify beta sitosterol oxides formed in thermal-oxidized vegetable oils which were heated at different temperatures and for varying time periods. Sitosterol oxidation is strikingly higher in sunflower oil relative to olive oil under all conditions of temperature and heating time. PMID- 16038956 TI - Tributyltin disturbs bovine adrenal steroidogenesis by two modes of action. AB - Tributyltin, an environmental pollutant, affected adrenal steroid hormone biosynthesis by two modes of action. Treatment of bovine adrenal cultured cells with 10-100 nM tributyltin for 48 h suppressed cortisol and androstenedione secretion, but induced the accumulation of 17alpha-hydroxyprogesterone and deoxycortisol, indicating that the P450(C21) and P450(11beta) activities were specifically suppressed. Direct inhibition of the enzymatic activities due to tributyltin was not observed in isolated organelles of untreated cells at concentrations less than 10 microM. Western blotting experiments using specific antibodies against steroidogenic enzymes showed that treatment with 1-100 nM tributyltin caused a decrease in cellular P450(C21) and P450(11beta) protein levels, and real-time PCR experiments showed that the decrease in protein content was attributable to decreases in mRNA of the enzymes. Tributyltin at concentrations higher than 100 nM suppressed all steroid biosynthesis in the adrenal cells. This suppression was closely correlated to the decrease in steroidogenic acute regulatory protein. Since nanomolar concentrations of tributyltin disturbed steroidogenesis in mammalian cells, there is the possibility that steroid hormone synthesis in polluted wild animals is affected by this compound. PMID- 16038957 TI - Gram-scale chromatographic purification of beta-sitosterol. Synthesis and characterization of beta-sitosterol oxides. AB - An effective purification method for beta-sitosterol was developed starting from a commercial source of a phytosterol mixture using preparative adsorption column chromatography. beta-Sitosterol (> or = 95% purity) was obtained on a gram-scale. Thus, the synthesis of six beta-sitosterol oxides, including 7alpha-hydroxy, 7beta-hydroxy, 5,6alpha-epoxy, 5,6beta-epoxy, 7-keto, and 5alpha,6beta dihydroxysitosterol, were successfully carried out. The spectral characteristics of all the synthetic intermediates and target compounds (approximately 95% purity) were well-documented. PMID- 16038958 TI - Therapeutic cloning: the ethical limits. AB - A brief outline of stem cells, stem cell therapy and therapeutic cloning is given. The position of therapeutic cloning with regard to other embryonic manipulations - IVF-based reproduction, embryonic stem formation from IVF embryos and reproductive cloning - is indicated. The main ethically challenging stages in therapeutic cloning are considered to be the nuclear transfer process including the source of eggs for this and the destruction of an embryo to provide stem cells for therapeutic use. The extremely polarised nature of the debate regarding the status of an early human embryo is noted, and some potential alternative strategies for preparing immunocompatible pluripotent stem cells are indicated. PMID- 16038959 TI - Epoxidation of the methamphetamine pyrolysis product, trans-phenylpropene, to trans-phenylpropylene oxide by CYP enzymes and stereoselective glutathione adduct formation. AB - Pyrolytic products of smoked methamphetamine hydrochloride are well established. Among the various degradation products formed, trans-phenylpropene (trans-beta methylstyrene) is structurally similar to styrene analogues known to be bioactivated by CYP enzymes. In human liver microsomes, trans-phenylpropene was converted to the epoxide trans-phenylpropylene oxide (trans-2-methyl-3 phenyloxirane) and cinnamyl alcohol. Incubation of trans-phenylpropene with microsomes in the presence of enzyme-specific P450 enzyme inhibitors indicated the involvement of CYP2E1, CYP1A2, and CYP3A4 enzymes. Both (R,R)-phenylpropylene oxide and (S,S)-phenylpropylene oxide were formed in human liver microsomal preparations. Enantiomers of trans-phenylpropylene oxide were stereoselectively and regioselectively conjugated in a Phase II drug metabolism reaction catalyzed by human liver cytosolic enzymes consisting of conjugation with glutathione. The structure of the phenylpropylene oxide-glutathione adduct is consistent with nucleophilic ring-opening by attack at the benzylic carbon. Exposure of cultured C6 glial cells to (S,S)-phenylpropylene oxide produced a cytotoxic response in a concentration-dependent manner based on cell degeneration and death. PMID- 16038960 TI - Dynamics of collinear contrast facilitation are consistent with long-range horizontal striate transmission. AB - It is well established that activity of striate neurons may be either facilitated or suppressed by visual stimuli presented outside of their classical receptive field (CRF) limits. Whilst two general mechanisms have been identified as candidates for these contextual effects; those based on extra-striate feedback and long-range horizontal striate connections; the physiological data supporting these models is both ambiguous and inconsistent. Here we investigate psychophysically the phenomenon of collinear facilitation, in which contrast detection thresholds for foveally presented Gabor stimuli are reduced via concurrent presentation of remote collinear flankers. Using backward noise masking, we demonstrate that the minimum exposure duration required to induce facilitation increases monotonically with greater target-flanker separation. The inferred cortical propagation velocities of this process (0.10-0.23 ms(-1)) closely correspond with depolarising activity observed to travel across striate cortex of several species. These dynamics strongly suggest that contrast facilitation is mediated via long-range horizontal striate connections. This conclusion complements a recent suggestion that collinear induced long-range suppressive dynamics depend on extra-striate feedback. PMID- 16038961 TI - Lignin modifying enzymes of Coriolopsis polyzona and their role in olive oil mill wastewaters decolourisation. AB - In order to decolourise olive oil mill wastewaters (OOMW) efficiently, production and differential induction of ligninolytic enzymes by the white rot Coriolopsis polyzona, were studied by varying growth media composition and/or inducer addition. Among various possible inducers, veratryl alcohol appeared to be the most efficient to enhance specific productions of lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase by a factor of 18.5, 20.8 and 55, respectively. Ligninolytic enzymes were better produced in glucose based medium with a low nitrogen level (2.2 mM) under O2 atmosphere. The addition of 5 mM veratryl alcohol resulted in a maximal production of LiP, whereas maximal MnP and laccase were obtained at 10 mM. LiP production was totally repressed in presence of 100 microM Mn2+. The extrapolation of these conditions on OOMW based media was carried out at different effluent dilutions and the possible role of the different ligninolytic enzymes in OOMW decolourisation was studied. A better effluent decolourisation was obtained under LiP induction condition (5 mM veratryl alcohol) than when LiP was repressed (100 microM Mn2+). Furthermore, high levels of laccase had a detrimental effect on OOMW decolourisation concomitant to the formation of soluble polymeric aromatic compounds. PMID- 16038962 TI - Notes on the models of cadmium kinetics. AB - This paper identifies several drawbacks in the models of absorption and desorption of cadmium presented by [Garcia-Gomez, C., Carbonell, G., 2004. Modelling the absorption and desorption of cadmium on paper pulp using kinetic approaches. Chemosphere 55, 869-878]. It then provides suggestions on improving the effectiveness of the model. PMID- 16038963 TI - Mass balance modelling of contaminants in river basins: a flexible matrix approach. AB - A novel and flexible approach is described for simulating the behaviour of chemicals in river basins. A number (n) of river reaches are defined and their connectivity is described by entries in an n x n matrix. Changes in segmentation can be readily accommodated by altering the matrix entries, without the need for model revision. Two models are described. The simpler QMX-R model only considers advection and an overall loss due to the combined processes of volatilization, net transfer to sediment and degradation. The rate constant for the overall loss is derived from fugacity calculations for a single segment system. The more rigorous QMX-F model performs fugacity calculations for each segment and explicitly includes the processes of advection, evaporation, water-sediment exchange and degradation in both water and sediment. In this way chemical exposure in all compartments (including equilibrium concentrations in biota) can be estimated. Both models are designed to serve as intermediate-complexity exposure assessment tools for river basins with relatively low data requirements. By considering the spatially explicit nature of emission sources and the changes in concentration which occur with transport in the channel system, the approach offers significant advantages over simple one-segment simulations while being more readily applicable than more sophisticated, highly segmented, GIS-based models. PMID- 16038964 TI - Voltage- and calcium-dependent inactivation in high voltage-gated Ca(2+) channels. AB - Calcium influx into cardiac myocytes via voltage-gated Ca channels is a key step in initiating the contractile response. During prolonged depolarizations, toxic Ca(2+) overload is prevented by channel inactivation occurring through two different processes identified by their primary trigger: voltage or intracellular Ca(2+). In physiological situations, cardiac L-type (Ca(V)1.2) Ca(2+) channels inactivate primarily via Ca(2+)-dependent inactivation (CDI), while neuronal P/Q (Ca(V)2.1) Ca(2+) channels use preferentially voltage-dependent inactivation (VDI). In certain situations however, these two types of channels have been shown to be able to inactivate by both processes. From a structural view point, the rearrangement occurring during CDI and VDI is not precisely known, but functional studies have underlined the role played by at least 2 channel sequences: a C terminal binding site for the Ca(2+) sensor calmodulin, essential for CDI, and the loop connecting domains I and II, essential for VDI. The conserved regulation of VDI and CDI by the auxiliary channel beta subunit strongly suggests that these two mechanisms may use a set of common protein-protein interactions that are influenced by the auxiliary subunit. We will review our current knowledge of these interactions. New data are presented on L-P/Q (Ca(V)1.2/Ca(V)2.1) channel chimera that confirm the role of the I-II loop in VDI and CDI, and reveal some of the essential steps in Ca(2+) channel inactivation. PMID- 16038965 TI - Protein profiling and identification of modulators regulated by human papillomavirus 16 E7 oncogene in HaCaT keratinocytes by proteomics. AB - OBJECTIVES: Viral oncogenes E6 and E7 are selectively retained and expressed in carcinoma cells infected with human papillomavirus type 16 and cooperated with each other in immortalization and transformation of primary keratinocytes. This study was performed to identify proteins to be bound or modulated by high risk HPV E7 oncogene by using a proteomics. METHODS: HaCaT normal keratinocyte was prepared to establish a stable cell line expressing E7. The E7-affinity column was also prepared to obtain E7-interacting proteins. In order to search the target molecules modulated by E7 expression, we used 2-dimensional electrophoresis and matrix-assisted laser desorption/ionization time of flight (MALDI/TOF) mass spectrometry. Pull down assay was also performed in order to confirm the E7-interacting proteins. RESULTS: We identified 28 spots that are modulated by E7 in HaCaT/E7 using 2-dimensional electrophoresis (2-DE) and MALDI/TOF mass spectrometry. Proteomics analyses showed that actin and leukocyte elastase inhibitor were down-regulated, whereas stress-induced phosphoprotein 1, CD2 binding protein 1, catalase, T-complex protein 1, Ku70-binding protein, heat shock 60 kDa protein 1, G1/S-specific cyclin E1 and peroxiredoxin 2 were up regulated. Western blot revealed that heat shock 60 kDa protein, catalase and peroxiredoxin 2 were also up-regulated. Pull down assay also showed that leukocyte elastase inhibitor (LEI) and Ku70-binding protein were bound to the E7 oncoprotein. By using E7-affinity column and 2-DE/MALDI-TOF, 22 spots were found to interact with E7 recombinant protein. MG11-like proteins, livin inhibitor-of apoptosis, protein serine kinase c17, CD2 binding protein 1, cyclin E1, TATA box binding protein-associated factor and uridine-cytidine kinase 2 were up-regulated by E7 oncogene and also bound to E7 oncoprotein. CONCLUSIONS: It is presumed that E7 can influence cell status by modulating the factors related to cell signaling, apoptosis and cell cycle regulation. PMID- 16038967 TI - Speech disruption under delayed auditory feedback in multilingual speakers. AB - The present study investigated the language familiarity hypothesis formulated by Mackay [(1970). How does language familiarity influence stuttering under delayed auditory feedback? Perceptual and Motor Skills, 30, 655-669] that bilinguals speak faster and stutter less under delayed auditory feedback (DAF) when speaking their more familiar language than a less familiar language. Thirty normally fluent native speakers of Dutch (17 males and 13 females, aged between 18;1 and 26;4 years) who were also proficient in French and English read meaningful and nonsense text under DAF in their mother tongue and in the two later acquired languages. The existence of a language familiarity effect was confirmed. The participants required significantly more time and showed significantly more speech disruptions under DAF in the later acquired languages than in the mother tongue, and reading time and number of speech disruptions was significantly higher for the nonsense texts than for the meaningful text for each of the three languages. An additional question addressed was whether or not there were any gender differences in the susceptibility to DAF. Results did not reveal a clear gender difference. EDUCATIONAL OBJECTIVES: The reader will be able to: (1) summarize the importance of language familiarity for the degree of speech disruption experienced by normally fluent multilingual speakers under delayed auditory feedback; and (2) describe gender differences in the susceptibility to delayed auditory feedback. PMID- 16038966 TI - Physical activity and decreased risk of clinical gallstone disease among post menopausal women. AB - BACKGROUND: Physical activity may play a role in preventing gallstone disease. METHODS: The activity/gallstone relationship was examined in post-menopausal women from the Study of Osteoporotic Fractures (SOF; 1986-1988), a prospective study of fracture risk factors in 8010 women (mean age = 71.1 years, SD = 4.9). RESULTS: Multivariate logistic regression indicated women in the lowest two quartiles of physical activity, according to questionnaire, had a 59% {OR = 1.59 (1.11-2.29), P = 0.02} and a 57% higher risk {OR = 1.57 (1.11-2.23), P = 0.01} of developing gallstone disease compared to women in the highest quartile of activity (PTrend = <0.0001). Additionally, this relationship was examined in a cohort of 182 post-menopausal women (mean age 74.2 years, SD = 4.1) who participated in a randomized controlled trial of a walking intervention. Women in the randomized clinical trial in the lowest tertile of physical activity determined by a physical activity monitor had a higher risk of developing gallstone disease than women in the highest tertile of physical activity, 13% {OR 1.13 (1.01-1.28), P = 0.05, PTrend = <0.04}. CONCLUSION: Physical activity appears to be inversely related to the development of gallstone disease in post menopausal women independent of body mass index. PMID- 16038968 TI - Influence of crosshead speed on micro-tensile bond strength of two-step adhesive systems. AB - OBJECTIVES: The purpose of this study was to determine the influence of crosshead speed on the micro-tensile bond strength of two separate adhesive systems to dentin. METHODS: The systems used were the Clearfil SE Bond (Kuraray Medical) and the Single Bond (3M ESPE) combined with a resin composite Clearfil AP-X (Kuraray Medical). Dentin surfaces of bovine madibular incisors were primed with self etching primer followed by air blowing for Clearfil SE Bond, or etched with phosphoric acid followed by rinsing with distilled water for Single Bond, and adhesive was applied. The resin composite was then built up in three layers and light activated. After 24 h storage in water, specimens were sectioned and trimmed to a cross-sectional area of 1 mm(2) and subjected to a micro-tensile bond-strength test. Ten samples per test group were tested at crosshead speeds of 0.5, 1.0, 5.0 and 10.0 mm/min. Micro-tensile bond-strength values (in MPa) were calculated from the peak load at failure divided by the specimen surface area. Two-way ANOVA was performed at the 0.05 probability level. RESULTS: The mean dentin bond strength at different crosshead speeds ranged from 34.6 to 37.1MPa for Clearfil SE Bond and from 44.3 to 50.4 MPa for Single Bond. There was no significant difference among the same adhesive systems with the different crosshead speeds tested. SIGNIFICANCE: The influence of the crosshead speed might be negligible when measuring micro-tensile bond strengths. PMID- 16038969 TI - Chemical aspects of self-etching enamel-dentin adhesives: a systematic review. AB - OBJECTIVES: The paper gives an overview on the components and the polymer chemical aspects of currently used self-etching enamel-dentin primers/adhesives. In addition, the contribution of new adhesives monomers and cross-linkers exhibiting enhanced hydrolytic stability than methacrylates to improve the performance of single-bottle adhesives is discussed. SOURCES: Information from original scientific papers or reviews about enamel-dentin adhesives, the patent literature concerning dental adhesives and manufacturer information of commercial self-etching adhesives were included in this review. DATA: The most efficient self-etching enamel-dentin adhesives are based on strongly acidic adhesive monomers, containing dihydrogenphosphate, phosphonic acids or carboxylic acid groups. Serious problems of single-bottle water-based, strongly acidic self etching enamel-dentin adhesives arise both from the hydrolytic instability of the methacrylate monomers used and the side reaction of the applied initiator components. CONCLUSIONS: The stability of the self-etching enamel-dentin adhesives can be improved by using new acrylic ether phosphonic acids or mono- or difunctional acrylamides, while more stable and compatible components have to be developed in the future. PMID- 16038970 TI - The effects of BIS-GMA and TEG-DMA on female mouse fertility. AB - OBJECTIVES: The current study evaluated the effect of bisphenol A glycerolate dimethacrylate (BIS-GMA) and triethyleneglycol dimethacrylate (TEG-DMA) on female mouse fertility. METHODS: Adult female mice were exposed to BIS-GMA or TEG-DMA (0, 25 and 100 microg/kg) intragastrically daily for 28 d and then mated with sexually mature untreated male mice and after mating their fertility was assessed. RESULTS: In females exposed to BIS-GMA at both doses significant increases in the total number of resorptions out of the total number of implantations were observed, with a significant increase in the number of animals with resorptions at the higher dose. Significant reductions in body weights and significant increases in ovary weights were also observed. Exposure to TEG-DMA at a dose of 100 microg/kg resulted in significant reductions in pregnancy rates and a significant increase in the total number of embryonal resorptions. Significant reductions in body and uterine weights were also observed in females exposed to TEG-DMA. SIGNIFICANCE: The results suggest that both BIS-GMA and TEG-DMA have reproductive toxic effects in female mice. PMID- 16038971 TI - Fit of metal ceramic crowns cast in Au-1.6 wt% Ti alloy for different abutment finish line curvature. AB - OBJECTIVES: The purpose of this study was to evaluate the fit of metal ceramic crowns cast in Au-1.6 wt% Ti alloy and investigate the effect of abutment finish line curvature on the fit of crowns. METHODS: Three types of finish line curvature abutments were prepared (1, 3 and 5mm-curvature). For each type of abutment, five metal ceramic crowns of the facial veneered type were fabricated, which were cast in Au-1.6 wt% Ti alloy. Used as controls, another fifteen specimens were made from a commercially available gold alloy. The fit was measured in the as-cast and after porcelain application. RESULTS: In the as-cast specimens, the greater the finish line curvature was, the larger the gaps exhibited at the mesial and distal margins of copings, compared with labial and lingual margins. The distal margin of copings for 5mm-curvature abutments showed the largest gap (35 (7) microm). After porcelain application, the greater was the finish line curvature, the larger the labial marginal gap became (mean 44, 34, 25 microm, respectively, for 5, 3, 1mm-curvature). However, there was no significant difference on marginal gaps between specimens of Au-1.6 wt% Ti alloy and control gold alloy. SIGNIFICANCE: This study indicated that the metal ceramic crowns cast in Au-1.6 wt% Ti alloy had equivalent accuracy to those that cast in control gold alloy, and the abutment finish line curvature had a significant effect on the marginal fit of metal ceramic crowns. PMID- 16038972 TI - The influence of underwater data transmission sounds on the displacement behaviour of captive harbour seals (Phoca vitulina). AB - To prevent grounding of ships and collisions between ships in shallow coastal waters, an underwater data collection and communication network (ACME) using underwater sounds to encode and transmit data is currently under development. Marine mammals might be affected by ACME sounds since they may use sound of a similar frequency (around 12 kHz) for communication, orientation, and prey location. If marine mammals tend to avoid the vicinity of the acoustic transmitters, they may be kept away from ecologically important areas by ACME sounds. One marine mammal species that may be affected in the North Sea is the harbour seal (Phoca vitulina). No information is available on the effects of ACME like sounds on harbour seals, so this study was carried out as part of an environmental impact assessment program. Nine captive harbour seals were subjected to four sound types, three of which may be used in the underwater acoustic data communication network. The effect of each sound was judged by comparing the animals' location in a pool during test periods to that during baseline periods, during which no sound was produced. Each of the four sounds could be made into a deterrent by increasing its amplitude. The seals reacted by swimming away from the sound source. The sound pressure level (SPL) at the acoustic discomfort threshold was established for each of the four sounds. The acoustic discomfort threshold is defined as the boundary between the areas that the animals generally occupied during the transmission of the sounds and the areas that they generally did not enter during transmission. The SPLs at the acoustic discomfort thresholds were similar for each of the sounds (107 dB re 1 microPa). Based on this discomfort threshold SPL, discomfort zones at sea for several source levels (130-180 dB re 1 microPa) of the sounds were calculated, using a guideline sound propagation model for shallow water. The discomfort zone is defined as the area around a sound source that harbour seals are expected to avoid. The definition of the discomfort zone is based on behavioural discomfort, and does not necessarily coincide with the physical discomfort zone. Based on these results, source levels can be selected that have an acceptable effect on harbour seals in particular areas. The discomfort zone of a communication sound depends on the sound, the source level, and the propagation characteristics of the area in which the sound system is operational. The source level of the communication system should be adapted to each area (taking into account the width of a sea arm, the local sound propagation, and the importance of an area to the affected species). The discomfort zone should not coincide with ecologically important areas (for instance resting, breeding, suckling, and feeding areas), or routes between these areas. PMID- 16038973 TI - The kinetic study of arginine kinase from the sea cucumber Stichopus japonicus with 5,5'-dithiobis-(2-nitrobenzoic acid). AB - The Stichopus japonicus arginine kinase (AK) is a significant dimeric enzyme. Its modification and inactivation course with 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) and the reactivation course of DTNB-modified AK by dithiothreitol were investigated on the basis of the kinetic theory of the substrate reaction during the modification of enzyme activity. The results show that the modification is a biphasic course while the inactivation is monophasic, with one essential reactive cysteine per subunit. The Cys274 (numbering from the Stichopus sequence) is exposed to DTNB and is near the ATP binding site. The modified AK can be reactivated by an excess concentration of dithiothreitol in a monophasic kinetic course. The presence of ATP or the transition-state analog markedly slows the apparent reactivation rate constant. The analog components, arginine-ADP-Mg2+ can induce conformational changes of the modified enzyme, but adding NO3- cannot induce further changes that occur with the native enzyme. The reactive cysteines' location and its role in the catalysis of AK are discussed. The results suggest that the cysteine may be located in the hinge area of the two domains of AK. The reactive cysteine of AK, which was proposed to be Cys274, may play an important role not in the binding of the transition-state analog but in the conformational changes caused by the transition-state analog. PMID- 16038974 TI - Increased CRF-like and NPY-like immunoreactivity in adult rats exposed to nicotine during adolescence: relation to anxiety-like and depressive-like behavior. AB - OBJECTIVE: Recently, animal models have been developed that demonstrate that adolescent nicotine exposure produces neurobehavioral changes which persist into adulthood. This study further examined the impact of adolescent nicotine exposure on anxiety-like and depressive-like behavior, as well as on levels of corticotropin-releasing factor (CRF) and neuropeptide Y (NPY) in this model. METHODS: Male adolescent rats (35-40 days old) were administered nicotine using Nicoderm CQ patches (Smith-Kline Beecham). Behavior in the elevated plus maze (EPM) and forced swim test (FST) was assessed 2-3 weeks after exposure ended. Brain levels of CRF and NPY were then assessed 5-6 weeks after behavioral tests were completed. In addition, blood and brain levels of nicotine resulting from nicotine treatment were examined. RESULTS: After 5 days of exposure to 5 mg/kg/day nicotine, blood levels of nicotine averaged 66+/-5 ng/ml and brain nicotine levels averaged 52+/-4 ng/g. Rats exposed to nicotine displayed an anxiety-like profile in the EPM (i.e., decreased time spent in the open arms) and an antidepressant-like profile in the FST (i.e., less time spent immobile). Rats exposed to nicotine also had increased hypothalamic and frontal cortical CRF, increased hypothalamic and hippocampal NPY, and a decreased ratio of NPY to CRF in the amygdala. CONCLUSIONS: This study demonstrates that adolescent nicotine exposure produces lasting increases in anxiety-like behavior and may reduce depressive-like behavior. These behavioral changes also occurred in concert with alterations in CRF and NPY systems. Thus, lasting neurobehavioral changes associated with adolescent nicotine exposure may be related to allostatic changes in stress peptide systems. PMID- 16038975 TI - Chemical interactions with snow: understanding the behavior and fate of semi volatile organic compounds in snow. AB - Snow plays an important role in providing atmospherically derived semi-volatile organic compounds (SVOCs) to regions of high latitude and altitude. The accumulated winter snowpack serves as a reservoir for SVOCs, which may then be released to arctic/alpine catchments during seasonal snowmelt or entrained into deeper layers of snow and ice. This paper provides a review of the occurrence of SVOCs in snow, exploring sampling methodologies and field measurements. Furthermore, chemical fate following snowfall and the propensity of SVOCs to undergo revolatilization with snow metamorphosis are examined along with air-snow partitioning and the role of physical parameters such as snow density and snow surface area in controlling vapor-sorbed levels. Snowmelt and firnification processes are described, and the latter are related to SVOC measurements made in deeper snow layers and glacial ice cores. Evidence is provided that suggests that those SVOCs that possess relatively higher snow interfacial/air partitioning coefficients (K(iasnow)) or lower Henry's Law constants may be more efficiently retained in snow, with implications for the occurrence of currently used pesticides in the temperate mountain snowpack. PMID- 16038976 TI - The accidental conservationist: William T. Hornaday, the Smithsonian bison expeditions and the US National Zoo. AB - In 1886 William T. Hornaday, the Chief Taxidermist of the US National Museum, led the "Smithsonian Institution Buffalo Outfit" to Montana. The American bison was fast heading toward extinction and the mission of the expedition was to locate those of the elusive animals that remained and obtain specimens for the scientific collection at the museum. The successful expedition produced the most complete scientific series, and the finest artistic grouping of taxidermied specimens of the American bison anywhere in the world. Haunted by the scattered skeletal remains of the millions of slaughtered bison that peppered the Eastern Montana Plains, Hornaday fought to establish the National Zoological Park, which would provide sanctuary for bison and other endangered species, and a captive breeding program, which would result in the eventual reintroduction of the American bison to the wild. PMID- 16038977 TI - Letter in response to A. Agrawal: 'CRP after 2004'. PMID- 16038978 TI - Syntheses, crystal structure and cytotoxicity of diamine platinum(II) complexes containing maltol. AB - The cationic complexes (1,2-diaminoethane)(maltolato)platinum(II) ([Pt(en)(ma)]+) and (1R,2R-1,2-diaminocyclohexane)(maltolato)platinum(II) ([Pt(R,R-DACH)(ma)]+) have been prepared and the structure of [Pt(R,R-DACH)(ma)]NO3 has been determined by single crystal X-ray diffraction. The geometry of the metal in [Pt(R,R DACH)(ma)]NO3 is essentially square planar and the maltolate ligand has a geometry similar to other chelate complexes involving this ligand. The cytotoxicities of the compounds have been assessed in the human cell lines HeLa and K562 and the IC50 values are approximately 32 microM in HeLa cells and 26 microM in K562 cells. In these cell lines the cytotoxicity of cisplatin is higher than the maltolate complexes by a factor of 2 to 3 whereas the cytotoxicity of carboplatin is lower than the maltolate complexes. PMID- 16038979 TI - DFT study of the V(IV)/V(V) oxidation mechanism in the presence of N hydroxyacetamide. AB - The oxidation mechanism of V(IV)/V(V) in the presence of N-hydroxyacetamide (acetohydroxamic acid, HL) in aqueous solution has been investigated using density functional theory (DFT) calculations aiming to contribute to the understanding of this process at a molecular level. The mechanism proposed involves formation of the *OH, *OOH, H2O2 radicals and complexes formed from the interaction of these species with VOL2 complex. The Gibbs free energy of each step of the mechanism has been evaluated. The solvation energy has been estimated by means of united atoms Hartree-Fock/polarizable continuum method (UAHF/PCM). The Gibbs free energy of the global reaction of the V(IV)/V(V) oxidation has been estimated and compared with the available experimental equilibrium constant. The difference between the calculated and experimental estimates for the reaction energy of the global equation is about 1.5 kcal mol(-1). The thermodynamic profile of the reaction mechanism has been provided and discussed in terms of the possible intermediates. The influence of the ligand and the reaction rate in terms of the steady-state approximation has been briefly discussed. PMID- 16038980 TI - Genital HIV-1 viral load is correlated with blood plasma HIV-1 viral load in Brazilian women and is reduced by antiretroviral therapy. AB - CONTEXT: Cervicovaginal secretions represent the primary vehicle in mother to infant and sexual HIV-1 transmission. Understanding the viral dynamics in this compartment is important to improve interventions to decrease HIV transmission. OBJECTIVES: To evaluate the correlation of HIV-1 blood plasma viral load and cervicovaginal HIV-1 viral load and the effect of antiretroviral therapy (ART) on cervicovaginal HIV-1 viral load. METHODS: A cross-sectional study among HIV-1 infected women recruited between February 2002 and January 2003 and a longitudinal study that included 11 women who initiated ART were performed. HIV-1 viral load was measured in the female genital tract and in blood plasma using the Nuclisens assay before and 1 month after ART introduction. RESULTS: HIV-1 viral load in cervicovaginal lavage was significantly correlated with HIV-1 blood plasma viral load (n = 27, Spearman rho = 0.73, p<0.001). In the longitudinal study, antiretroviral therapy was associated with a reduction of 1.44 log10 on genital HIV-1 viral load (95% CI = 0.57-2.30, p = 0.004, Student's t-test). CONCLUSIONS: HIV-1 viral load in female genital secretions is correlated with blood plasma HIV-1 viral load. Antiretroviral therapy substantially reduces HIV-1 viral load in the female genital tract. PMID- 16038981 TI - Involving the pharmaceutical and biotech communities in medication development for substance abuse. AB - Pharmacotherapy as adjunctive treatment is an integral part of the strategy for treating substance abuse. Although there are several approved drugs for the treatment of opioid, alcohol, and nicotine dependence, the pharmaceutical industry, for a variety of reasons, has been reluctant to enter this area to develop medications for substance abuse indications. Therefore, in 1990, a Medication Development Program was established by NIDA to carry out and assist in stimulating development of new pharmacotherapies. It is vital for NIDA to provide clear leadership and establish a collaborative working relationship with the pharmaceutical industry, providing scientific, development, and financial assistance, depending on the size, resources, and expertise of the company. An important NIDA role in this effort is setting standards, such as establishing Target Product Profiles (TPPs), predictive decision trees for selection of clinical candidates, and animal models to evaluate safety and potential effectiveness prior to human studies. NIDA can further establish standards for clinical studies, including Proof of Concept (PoC), Phase 2 (or Learning) trials to establish initial proof of safety and effectiveness, and Phase 3 (or Confirming) trials to validate Phase 2 findings. NIDA and other government agencies need to work to improve industry incentives to participate in medication development for substance abuse. Specific incentives, such as market exclusivity and patent extension, as provided in BioShield and pediatric drug legislation, should be strongly considered. NIDA can further assist industry to navigate the regulatory and, if needed, controlled substance scheduling processes, by establishing a true Federal partnership between NIDA, FDA, and DEA. PMID- 16038982 TI - Would modulation of intracellular Ca2+ be antiarrhythmic? AB - Under several types of conditions, reversal of steps of excitation-contraction coupling (RECC) can give rise to nondriven electrical activity. In this review we explore those conditions for several cardiac cell types (SA, atrial, Purkinje, ventricular cells). We find that abnormal spontaneous Ca2+ release from intracellular Ca2+ stores, aberrant Ca2+ influx from sarcolemmal channels or abnormal Ca2+ surges in nonuniform muscle can be the initiators of the RECC. Often, with such increases in Ca2+, spontaneous Ca2+ waves occur and lead to membrane depolarizations. Because the change in membrane voltage is produced by Ca2+-dependent changes in ion channel function, we also review here what is known about the molecular interaction of Ca2+ and several Ca2+-dependent processes, including the intracellular Ca2+ release channels implicated in the genetic basis of some forms of human arrhythmias. Finally, we review what is known about the effectiveness of several agents in modifying such Ca2+-dependent arrhythmias. PMID- 16038983 TI - FlexStation examination of 5-HT3 receptor function using Ca2+ - and membrane potential-sensitive dyes: advantages and potential problems. AB - The FlexStation is a 96 or 384 fluorescent plate reader with the capability of adding solutions during readings; it therefore has the potential to provide high throughput analyses of the functional characteristics of neurotransmitter-gated ion channels that can be examined using changes in fluorescence. The 5-HT3 receptor is one such protein, as its activation results in a change in membrane potential due to the opening of a Ca2+ -permeant, cation-selective channel; it can therefore be studied using both Ca2+ - and membrane potential-sensitive fluorescent dyes. Here we have used the FlexStation to examine the function of recombinant 5-HT3 receptors expressed in HEK293 cells using both these classes of dye. The results show that the pharmacological characteristics of the receptor obtained using the FlexStation is similar to those reported using other functional methods, although caution must be applied when using the membrane potential dye as large changes in membrane potential can yield inaccurate EC50s. Modifying the constituents of the buffer, however, so that the change in membrane potential was reduced, yielded EC50 values that were similar to previously reported data. We conclude that the FlexStation is a useful tool for high throughput studies when examining the function of neurotransmitter receptors that result in either a change in Ca2+ concentration or membrane potential. PMID- 16038984 TI - The relationship between job-related burnout and depressive disorders--results from the Finnish Health 2000 Study. AB - BACKGROUND: Depression and burnout are common health problems in working populations today. They appear to be interrelated, and the need for their differential diagnosis has been highlighted in many reviews. We analysed the overlap of job-related burnout and depressive disorders, i.e., major depressive disorder, dysthymia, and minor depressive disorder. METHODS: We used the population-based 'Health 2000 Study' in Finland. Our nationally representative sample comprised 3276 employees aged 30-64 years. Burnout was assessed with the Maslach Burnout Inventory-General Survey. Diagnoses of depressive disorders were based on the Composite International Diagnostic Interview. RESULTS: Burnout and depressive disorders were clearly related. The risk of depressive disorders, especially major depressive disorder (12-month prevalence), was greater when burnout was severe. Half of the participants with severe burnout had some depressive disorder. Those with a current major depressive episode suffered from serious burnout more often than those who had suffered a major depressive episode earlier. LIMITATIONS: This study was cross-sectional. CONCLUSIONS: The concepts of burnout and depression complement each other and cover partly overlapping phenomena. Depressive disorders are related to job-related burnout, particularly when it is severe. A current major depressive episode is likely to be associated with the experience of burnout. When encountering working patients, it is recommended to assess both the occurrence of burnout and of depressive disorders. PMID- 16038985 TI - Presence of the P2X(7) purinergic receptor on immune cells that invade the rat endometrium during oestrus. AB - Eosinophils, macrophages and other leucocytes invade the uterine endometrium during oestrus and play a role in the tissue remodeling and immune responses that occur prior to implantation of the fertilized ovum. Adenosine 5'-triphosphate (ATP) and its metabolites influence uterine function via ATP receptors. In this study, we investigated the presence and localisation of the P2X(7) nucleotide receptor in the cells that infiltrate the uterine endometrium of adult female rats during oestrus at the electron microscope level, using gold-silver pre embedding immunocytochemical techniques. P2X(7) receptor expression was found in the cytoplasm and the cell membrane of eosinophils, macrophages and fibroblasts in the endometrium during oestrus. These results suggest that ATP-mediated responses may be important in uterine preparation and remodeling before implantation and that this may involve several types of cells. In particular, the presence of P2X(7) receptors on endometrial stromal cells may indicate their involvement in apoptosis and immune and inflammatory responses. PMID- 16038986 TI - Genetic variation in the interleukin-8 gene promoter and vaginal concentrations of interleukin-8 are not associated with bacterial vaginosis during pregnancy. AB - Interleukin-8 (IL-8) may play a role in the activation of the vaginal immune system during bacterial vaginosis. However, contradictory results were obtained regarding the involvement of IL-8 in the immunological response during bacteria vaginosis. These apparently contradictory results could be due to different genetic variations of the study groups. Since some gene polymorphisms may affect the level of IL-8 production, the aim of this study was to determine whether the frequency IL-8 promoter alleles and levels of IL-8 in vaginal fluid are associated with bacterial vaginosis during pregnancy. Genotyping for IL-8 polymorphisms in the promoter region of the gene was performed in 34 pregnant women with asymptomatic bacterial vaginosis matched for gestational age with 38 pregnant women without vaginosis. Additionally, vaginal IL-8 levels were assayed by the dual monoclonal antibody sandwich enzyme-linked immunosorbent assay technique. The frequencies of the three polymorphisms were not significantly different between control women and women with bacterial vaginosis. In addition, there was no linkage disequilibrium between the polymorphisms. Furthermore, there was no statistical difference in median vaginal levels of IL-8 between both groups. Neither the frequencies of IL-8 polymorphic alleles nor levels of IL-8 in vaginal fluid were associated with bacterial vaginosis. PMID- 16038987 TI - Age has a similar influence on the susceptibility to NMDA antagonist-induced neurodegeneration in most brain regions. AB - NMDA antagonists are of potential therapeutic benefit for several conditions. However, their ability to produce neurotoxicity and psychosis has hampered their clinical use. A better understanding of these side effects and the mechanism underlying them could result in their safer use and in improving our understanding of psychotic illnesses. By disinhibiting certain multisynaptic circuits, moderate doses of NMDA antagonists produce reversible neurotoxicity in the retrosplenial cortex in rats older than 1 month. Higher doses of these same agents result in the death of neurons in the retrosplenial cortex and several other brain regions. It is unknown whether susceptibility to this irreversible neurodegeneration has a similar age dependency profile. We, therefore, examined the sensitivity of rats of various ages (PND20-60) to the irreversible neurodegenerative effect of the selective NMDA antagonist, MK-801. Quantification of the severity of neurodegeneration with stereology revealed that the retrosplenial cortex, induseum griseum, and dentate gyrus had decreasing amounts of damage with decreasing age and onset of sensitivity around PND30. The piriform cortex also displayed a decreased amount of degeneration in younger age groups. However, a low level of degeneration continued to occur in the posterior piriform cortex in the PND20-25 animals. The stage of degeneration appeared to be more advanced, suggesting that these neurons were dying by a different mechanism. We conclude that for most neuronal populations, susceptibility to the irreversible and reversible neurodegenerative effects of NMDA antagonists has a similar age dependency profile, consistent with the proposal that the same disinhibitory mechanism underlies both neurotoxicities. PMID- 16038988 TI - Nitric oxide synthases II and III and vascular endothelial growth factor are up regulated in sciatic nerve after spiral cuff electrode implantation. AB - Nerve cuff electrodes, commonly used in functional electrical stimulation systems, induce local morphological changes that can affect nerve functionality. Nitric oxide (NO) and vascular endothelial growth factor (VEGF) have both neural and vascular effects. We investigated the time-dependent regulation of nitric oxide synthases (NOS) and of VEGF after implantation of spiral cuff electrode around rat sciatic nerve. NOSIII as well as VEGF were up-regulated in both epineurial and endoneurial compartments in cuff-implanted animals along with microvascular changes. Our results suggest that VEGF and NO are implied in morphological and functional alterations occurring in the early time after cuff implantation. PMID- 16038989 TI - Perinatal asphyxia, hyperthermia and hyperferremia as factors inducing behavioural disturbances in adulthood: a rat model. AB - Alertness was studied in adult male Wistar rats after neonatal critical anoxia applied under three different thermal conditions: (i) at physiological neonatal body temperature of 33 degrees C, (ii) at body temperature elevated to 37 degrees C, and (iii) at body temperature elevated to 39 degrees C (both during anoxia and for 2 h postanoxia). To elucidate the effect of iron-dependent postanoxic oxidative damage to the brain, half of the group (iii) was injected with deferoxamine, a chelator of iron. Postanoxic behavioural disturbances were recorded in open-field, elevated plus-maze, and sudden silence tests when the rats reached the age of 4 month. Moreover, spontaneous motor activity of the rats was recorded radiotelemetrically in their home-cages. Both open-field stress induced and spontaneous motor activity were reduced in rats subjected to neonatal anoxia under hyperthermic conditions. In contrast, these rats were hyperactive in the plus-maze test. Both the plus-maze and sudden silence tests revealed that these rats show reduced alertness to external stimuli signalling potential dangers. The behavioural disturbances were prevented by the body temperature of 33 degrees C and by postanoxic administration of deferoxamine. These data support the conclusion that permanent postanoxic behavioural disturbances are due to iron dependent oxidative damage to the brain, which can be prevented by the reduced neonatal body temperature. PMID- 16038990 TI - Cingulate cortex activation and competing responses: the role of preparedness for competition. AB - Regional cerebral blood flow (rCBF) was studied in a task, where a preparatory stimulus (S1) cued for an imperative second stimulus (S2) which was associated with a response. Two preparatory stimuli cued unequivocally each for one response. In contrast, a third preparatory stimulus cued for two response alternatives which appeared for the same ratio (each in 50% of all trials) introducing response competition. In a first experimental condition, non arbitrary, unambiguous stimuli were used as S1 to enable the subjects to prepare their responses. In a second and third scan, arbitrary preparatory stimuli were used during different stages of awareness for the S1-S2 association. Subjects performed this task "naive" without knowledge about the S1-S2 association and also in an experimental condition being aware of the S1-S2 association. Button presses after unambiguous, non-arbitrary preparatory stimuli activated the right middle frontal gyrus and inferior parietal lobe if S1 was associated with a definite response. When the subjects did not know the S1-S2 relation, left prefrontal cortex activation was associated with trials including definite responses. Performing the same S1-S2 response condition after subjects knew their relation right prefrontal and left parietal areas became additionally engaged. However, in the first experimental condition using unambiguous, non-arbitrary stimuli and in the third, "aware" experimental condition when S1 was coupled with two response alternatives, the anterior cingulate cortex was activated. As these experimental conditions have in common, that the preparatory stimulus shares information about the upcoming competing response alternatives they highlight the evaluative role of the anterior cingulate cortex for competing actions. PMID- 16038991 TI - Diabetes mellitus in the elderly: diagnostic features. AB - The new diagnostic criteria of the diabetes do not consider the age of the subjects. It has been demonstrated that the fasting glucose values as well as the glycemia levels after oral glucose load increase with age. Therefore, it is necessary to evaluate whether the new diagnostic criteria are valid for the elderly subjects also. In spite of the findings that among the subjects with fasting glycemia of 126-140 mg/ml subsequent controls after long times often revealed the presence of impaired fasting glucose (IFG) and numerous cases with IFG displayed values <110 mg/dl after longer times, the present authors accept the new diagnostic criteria even for the elderly subjects. However, they emphasize the importance of distinguishing diabetes in the elderly in subclasses, such as clinically manifest diabetes, mild diabetes and diabetic glucose tolerance (DGT) type, in order to establish the early diagnosis of diabetes, and therefore, to prevent the chronic vascular complications. PMID- 16038992 TI - Incremental prognostic value of stress echo positivity in the left anterior descending coronary artery territory. AB - BACKGROUND: Dipyridamole stress echo (DSE) positivity is usually titrated according to presence and severity of the induced wall motion abnormalities. The purpose of our study is to assess whether the location of DSE positivity might add to prognostic stratification. METHODS: The study enrolled 112 patients with known or suspected coronary artery disease (CAD) and without a history of prior myocardial infarction. They were consecutively submitted to DSE (0.84 mg/Kg in 10 min followed by atropine administration, when needed) and coronary angiography (within a 15+/-7 day period). End points at follow-up were cardiac death and acute coronary syndromes. RESULTS: Twenty-seven patients had a negative, and 85 a positive DSE (47 in the LAD territory). Angiographically assessed CAD was present in 82 patients (LAD in 59). At a follow-up of 9+/-7 months, there were 28 events of cardiac deaths or acute coronary syndromes. Event-free survival was lower in patients with positive DSE (any location) compared to those with negative DSE (47% vs 89%, p=0.003). In the subset with positive DSE, event-free survival was lower in patients with wall motion abnormalities in the LAD territory compared to those with dysfunction in the left circumflex and right coronary artery territory (31% vs 72%, p=0.00012 ). At multivariate analysis, stress echo positivity in the LAD territory was independently associated with increased risk (HR: 9.51, CI: 1.61 to 56.11, p<0.013) and outperformed angiographically assessed LAD stenosis (HR: 0.36, CI: 0.06 to 2.24, p: NS). CONCLUSIONS: In CAD patients, wall motion abnormalities in the LAD territory during DSE identified a higher risk subgroup. Functional stress echo positivity on the LAD territory overcame the prognostic impact of an anatomic, angiographically assessed LAD disease. PMID- 16038993 TI - Assessment of right and left ventricular function by tissue Doppler echocardiography in patients after biventricular repair of pulmonary atresia with intact ventricular septum. AB - BACKGROUND: Survival of patients with PAIVS has improved significantly with adoption of a selective management approach based on RV and coronary arterial anatomy. We sought to determine the right (RV) and left ventricular (LV) function of patients with pulmonary atresia and intact ventricular septum (PAIVS) long term after biventricular repair. METHODS: The regional ventricular function of 22 patients (11 males) aged 12.8+/-5.6 years with PAIVS after biventricular repair was assessed by tissue Doppler echocardiography (TDE). The results were compared to those of 22 age-matched controls. RESULTS: When compared with controls, patients had significantly reduced myocardial tissue velocities, systolic and diastolic strain rate, and peak systolic strain in the basal, mid-, and apical segments of the RV free wall, interventricular septum and LV lateral wall (all with p<0.01). Heterogeneity of systolic and diastolic strain rate was observed in the RV free wall and interventricular septum in patients but not in controls. CONCLUSION: Impairment of RV and LV regional longitudinal myocardial function occurs in patients with PAIVS late after biventricular repair. PMID- 16038994 TI - Prosthetic heart valve thrombosis: pathogenesis, diagnosis and management. AB - Thrombosis of a prosthetic valve is one of the most severe complications of cardiac valve replacement. The incidence is 0.5% in the aortic and mitral positions and up to 20% in the tricuspid position. The presenting clinical picture ranges from the absence of symptoms to cardiogenic shock. The traditional treatment of this complication has been emergency surgery, but thrombolysis, which has been available for many years, is being considered as the first line of treatment more and more every day. The pathogenesis, diagnosis and treatment of thrombosis of prosthetic heart valves are reviewed here. Thrombolysis, which has an efficacy of over 80%, is emphasized in this review. Embolic complications associated with this therapeutic approach remain a great concern with rates of 3% to 10%, and some authors reporting rates up to 20%. PMID- 16038995 TI - Opposite effects of insulin-like molecules and leptin in coronary heart disease of type 2 diabetes Preliminary data. AB - INTRODUCTION: Leptin and insulin have been reported to be risk factors for coronary heart disease (CHD) in the general population, but their role in type 2 diabetes still remains unclear. MATERIALS AND METHODS: The role of leptin and insulin upon CHD in type 2 diabetes was assessed in 154 patients, aged 31-77 years, who were treated with oral anti-diabetic agents. Multivariate logistic regression analyses were used with CHD (an established history of CHD or an abnormal treadmill test) as dependent, and leptin, insulin and potential confounders as independent variables. RESULTS: Endogenous insulin was significantly associated with CHD in a model controlling for gender, age, duration of diabetes, body mass index, smoking and leptin (Odds ratio 1.45 per decile, 95% confidence interval 1.11-1.90). Improving control for confounding by replacing body mass index by subcutaneous fat (CT-measured at the L4-L5 level) and height in this model, resulted in a significant negative association between leptin and CHD (OR 0.60, 95% CI 0.37-0.96). DISCUSSION: Leptin might have a beneficial effect on CHD in type 2 diabetes, probably by counteracting the effect of insulin-like molecules or insulin resistance. The effect was elucidated only after careful control for confounding by subcutaneous fat, the main source of leptin production. PMID- 16038996 TI - From chemical structure to environmental biosynthetic pathways: navigating marine invertebrate-bacteria associations. AB - Many natural products from marine invertebrates are promising drugs or drug leads but the problem of supply has delayed the development of these agents. Bacteria living in complex associations with animals are often proposed to be the real producers of "invertebrate" metabolites. Recent advances in biosynthetic pathway discovery from these symbiotic bacteria herald a new era in which biosynthetic genes will be cloned rapidly to provide promising molecules from marine invertebrates. PMID- 16038997 TI - Simple but predictive protein models. AB - The traditional approach to computational biophysics studies of molecular systems is brute force molecular dynamics simulations under the conditions of interest. The disadvantages of this approach are that the time and length scales that are accessible to computer simulations often do not reach biologically relevant scales. An alternative approach, which we call intuitive modeling, is hypothesis driven and based on tailoring simplified protein models to the systems of interest. Using intuitive modeling, the length and time scales that can be achieved using simplified protein models exceed those of traditional molecular dynamic simulations. Here, we describe several recent studies that signify the predictive power of simplified protein models within the intuitive-modeling approach. PMID- 16038998 TI - High level expression of surface glycoprotein of rabies virus in tobacco leaves and its immunoprotective activity in mice. AB - A synthetic gene coding for the surface glycoprotein (G protein) of rabies virus was strategically designed to achieve high-level expression in transgenic plants. The native signal peptide was replaced by that of the pathogenesis related protein, PR-S of Nicotiana tabacum. An endoplasmic reticulum retention signal was included at C-terminus of the G protein. Tobacco plants were genetically engineered by nuclear transformation. Selected transgenic lines expressed the chimeric G protein at 0.38% of the total soluble leaf protein. Mice immunized intraperitoneally with the G protein purified from tobacco leaf microsomal fraction elicited high level of immune response as compared to the inactivated commercial viral vaccine. The plant-derived G protein induced complete protective immunity in mice against intracerebral lethal challenge with live rabies virus. The results establish that plants can provide a safe and effective production system for the expression of immunoprotective rabies virus surface protein. PMID- 16038999 TI - Genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatography. AB - An engineered horseradish peroxidase isozyme C (HRP C) gene was constructed by the addition of a 6xArg fusion tail to 6xHis-HRP C by the PCR strategy. The 6xHis 6xArg-HRP C cDNA was expressed in the Sf9 insect cell line from Spodoptera frugiperda infected with Autographa californica nuclear polyhedrosis virus. The recombinant peroxidase isoelectric point was 9.5 as judged by isoelectric focusing and was purified directly from the culture medium at day-6 post infection by cation-exchange chromatography or immobilised metal ion-affinity chromatography. While the former technique gave a yield of 98.5% with a purification factor of 130, the latter gave only a 68% yield with a purification factor of 140. Results obtained provide evidence that the poly-Arg tag is more effective than the poly-His tag for peroxidase purification from a baculovirus expression system. PMID- 16039000 TI - Vaccinia virus K1L protein mediates host-range function in RK-13 cells via ankyrin repeat and may interact with a cellular GTPase-activating protein. AB - The K1L protein of vaccinia virus is required for its growth in certain cell lines (RK-13 and human). The cowpox host-range protein CP77 has been shown to complement K1L function in RK-13 cells, despite a lack of homology between the two proteins except for ankyrin repeats. We investigated the role of ankyrin repeats of K1L protein in RK-13 cells. The growth of a recombinant vaccinia virus, with K1L gene mutated in the most conserved ankyrin repeat, was severely impaired. Infection with the mutant virus caused shutdown of cellular and viral protein synthesis early in infection. We also investigated the interaction of K1L protein with cellular proteins and found that K1L interacts with the rabbit homologue of human ACAP2, a GTPase-activating protein with ankyrin repeats. Our result suggests the importance of ankyrin repeat for host-range function of K1L in RK-13 cells and identifies ACAP2 as a cellular protein, which may be interacting with K1L. PMID- 16039001 TI - Polyplex Nanogel formulations for drug delivery of cytotoxic nucleoside analogs. AB - Hydrophilic nanosized particles consisting of the cross-linked cationic polymer network (Nanogels) are suggested as a drug delivery system for nucleoside analog 5'-triphosphates, an active form of cytotoxic anticancer drugs. Preparation, properties, and cellular effects of several polyplex Nanogel formulations with the 5'-triphosphate of cytotoxic 5-fluoroadenine arabinoside (fludarabine) (FATP) were examined and discussed here. The polyplexes have formed spontaneously by mixing solutions of FATP and Nanogels because of ionic interactions between protonated polyethylenimine (PEI) chains in Nanogel network with polyphosphate groups of the drug. Subsequent compaction of the flexible Nanogel network has resulted in an encapsulation of the FATP/PEI complex in a dense core surrounded by hydrophilic poly(ethylene glycol) (PEG) envelope. This structure has provided a sustained release of the drug, as well as an efficient protection of FATP against enzymatic degradation. The drug loading could reach up to 33% by weight of the drug-Nanogel formulation. In vitro 35% of loaded drug has released from Nanogel formulations during the first 24 h, and a slower additional release was observed during the next 2 days. Nanogels have protected 90% of the encapsulated FATP from enzymatic dephosphorylation during the first 60 min of incubation in vitro. The drug-Nanogel formulation compared to the drug has demonstrated a significantly enhanced cytotoxicity in cultured cancer cells. Cancer cell targeting molecules, such as folate, could be easily attached to Nanogels and this modification has resulted in a strong 10-fold increase of the carrier's internalization in human breast carcinoma MCF-7 cells. Moreover, transcellular transport of the folate-Nanogel polyplexes was found to be 4 times more effective compared to the drug alone using Caco-2 cell monolayers as an in vitro intestinal model. The data demonstrate that this carrier-based approach to delivery of cytotoxic drugs may enhance tumor specificity and significantly reduce side effects related to systemic toxicity usually observed during cancer chemotherapy. PMID- 16039002 TI - Translations of health sector SWAps--a comparative study of health sector development cooperation in Uganda, Zambia and Bangladesh. AB - INTRODUCTION: Sector-wide approach, or SWAp, is an increasingly popular approach to development cooperation for many bilateral and multilateral development agencies. While much has been published about the SWAp model itself, there is still limited research conducted on how it is applied in practice. This study aims at showing how the SWAp is translated as it is adopted in different local settings and it shows how these translations trigger changed practice in different ways. METHODOLOGY: A three-layered comparative study of exemplary texts, country policies and interviews, was conducted in Uganda, Zambia and Bangladesh. An international literature review, comprising research articles, consultancy reports and government policy documents was undertaken. Also, a total of 64 interviews were conducted with representatives from the government, development partners and NGOs. Finally, 18 policy and donor meetings at national and district levels were attended and observed. RESULTS: The study showed that the definitions of the SWAp model are general and vague in character. Furthermore, it was observed that in all three countries, no serious effort has been made to define SWAp in country policies. Finally, the study revealed that partners in development translate SWAp differently in daily practice. The variations in understandings were found both between and within countries. CONCLUSIONS: The SWAp is a clear label with unclear content. The meaning of SWAp is formed by the recipient government and its development partners through an active process of translation as the SWAp is adopted in a specific context. The introduction of the SWAp has limited potential to transform standards for how development aid should be pursued and practiced as much of the administrative structures in each country are still formed in line with a project focused approach. It is concluded that in order to create change, partners in development must work, not only on defining SWAp, but also on influencing the context surrounding the SWAp. PMID- 16039003 TI - The effect of the Americans with Disabilities Act on economic well-being of men with disabilities. AB - This paper estimates the effect of the Americans with Disabilities Act (ADA) on the economic well-being of working-age (20-62) men with disabilities by fitting a fixed-effects model to the U.S. Survey of Income and Program Participation (SIPP) panel data. This paper constructs a new and alternative measure of disability utilizing rich information on limitations to functional activities (FA), activities of daily living (ADL), and instrumental activities of daily living (IADL). The new objective measure of disability identifies a larger group of people as being disabled than conventional self-reported measure. Probability of employment for disabled men fell 5.8% compared to nondisabled men significantly at the 5% level. Log-real-wages of men with disabilities fell 5.3% relative to men without disabilities significantly at the 10% level. PMID- 16039004 TI - The splicing machinery is a genetic modifier of disease severity. AB - Disease severity correlates with the level of correctly spliced RNA transcribed from genes carrying splicing mutations and with the ratio of alternatively spliced isoforms. Hence, a role for splicing regulation as a genetic modifier has been suggested. Here we discuss recent experiments that provide direct evidence that changes in the level of splicing factors modulate the splicing pattern of disease-associated genes. Importantly, modulation of the splicing pattern led to regulation of the protein function and modification of disease severity. PMID- 16039005 TI - Intergenomic conflict revealed by patterns of sex-biased gene expression. AB - Intergenomic conflict can affect the distribution of genes across eukaryotic genomes. Because the phenotypic optima of males and females often differ, the fitness consequences of newly arisen alleles might not be concordant between the sexes and can be sexually antagonistic--genetic variants favored in one sex are deleterious in the other. In this article, we demonstrate that previously unexplained patterns of sex-biased gene expression in Drosophila melanogaster might have evolved by sexual antagonism, and that the majority of sex-biased expression is due to adaptive changes in males, implying that males experience stronger selection than females. PMID- 16039006 TI - Transfection of tyrosine kinase deleted FGF receptor-1 into rat brain substantia nigra reduces the number of tyrosine hydroxylase expressing neurons and decreases concentration levels of striatal dopamine. AB - The effects of HSV-1 amplicon and polyethyleneimine (PEI)-mediated transfection of dominant negative FGF receptor-1 mutant FGFR1(TK-) into the rat brain substantia nigra (SN) were examined in vivo to model the reduced FGF signaling documented to occur in Parkinson's disease. The number of SN neurons that expressed tyrosine hydroxylase (TH) was significantly reduced following HSV-1 FGFR1(TK-) intranigral delivery and similar changes were observed after PEI mediated FGFR1(TK-) transfections. Further, we also observed a significantly lower striatal dopamine content following the PEI transfection of FGFR1(TK-). Thus, we conclude that reduced FGF signaling in the SN of Parkinsonian patients could play a role in the impaired dopaminergic transmission associated with the degenerative disease. PMID- 16039007 TI - Alpha1-adrenoreceptor in human hippocampus: binding and receptor subtype mRNA expression. AB - Alpha1-adrenoreceptors (AR), of which three subtypes exist (alpha1A-, alpha1B- and alpha1D-AR) are G-protein-coupled receptors that mediate the actions of norepinephrine and epinephrine both peripherally and centrally. In the CNS, alpha1-ARs are found in the hippocampus where animal studies have shown the ability of alpha1-AR agents to modulate long-term potentiation and memory; however, the precise distribution of alpha1-AR expression and its subtypes in the human brain is unknown making functional comparisons difficult. In the human hippocampus, 3H-prazosin (alpha1-AR antagonist) labels only the dentate gyrus (molecular, granule and polymorphic layers) and the stratum lucidum of the CA3 homogeneously. Human alpha1A-AR mRNA in the hippocampus is observed only in the dentate gyrus granule cell layer, while alpha1D-AR mRNA expression is observed only in the pyramidal cell layers of CA1, CA2 and CA3, regions where 3H-prazosin did not bind. alpha1B-AR mRNA is not expressed at detectable levels in the human hippocampus. These results confirm a difference in hippocampal alpha1-AR localization between rat and humans and further describe a difference in the localization of the alpha1A- and alpha1D-AR mRNA subtype between rats and humans. PMID- 16039008 TI - Snail peptide expression pattern in the nervous system of the medicinal leech. AB - Distribution of neurons immunopositive to antibodies against the "command neuron peptides" (CNPs) encoded by the snail Helix Command-Specific 2 (HCS2) gene was investigated in the nervous system of medicinal leech Hirudo. Immunopositive neurons were found in the leech segmental ganglia, brain and tail ganglionic masses, and peripheral ganglia. The CNPs immunopositive fibers were observed in neuropils of all ganglia and in some nerves. The role of CNPs immunopositive cells in animal behavior and the putative functions of the CNPs neuropeptide family are discussed. PMID- 16039010 TI - Oral temozolomide in heavily pre-treated brain metastases from non-small cell lung cancer: phase II study. AB - INTRODUCTION: The primary tumour type most likely to metastasize to the brain is lung cancer. In heavily pre-treated patients, limited therapeutic option is available and the results of availability therapies reported in literature are disappointing. The present phase II study was designed to assess the efficacy and safety of temozolomide (TMZ) as palliative treatment for brain metastases (BrM) in NSCLC patients pre-treated with WBRT and at least one line of chemotherapy for metastatic brain disease. MATERIAL AND METHODS: Temozolomide was administered orally at 150 mg/mq/day for five consecutive days for the first cycle, doses were increased to 200 mg/mq/day for 5 days every 28 days for subsequent cycles if no grade 3/4 haematological toxicity was observed. Eligibility criteria included cytological or histological confirmed NSCLC; BrM, recurrent or progressing after WBRT and at least one line of chemotherapy. A total of 30 consecutive patients entered the study and received the allocated treatment. RESULTS: Three patients (10%) achieved an objective response (OR) of BrM with two complete remission. Stable disease and progressive disease were achieved in 3 (10%) and 24 patients (80%), respectively. A correlation between response to TMZ and sensitivity to the previous first line chemotherapy was reported. Time to progression and overall survival were examined both for responder patients and for all included patients. For long-term survivors, we considered the patients who survived >12 months after the start of TMZ. According to this definition, three patients resulted long-term survivors: 2 with OR and 1 with stable brain disease. No grades 3 or 4 toxicity occurred. The total of treatment-related adverse events were mild or moderate (G1 2) in intensity. No patients discontinued TMZ as a result of treatment-related toxicity. DISCUSSION: The results of the present trial clearly demonstrates that TMZ is active and safe in BrM NSCLC patients previously treated with WBRT and at least one line of chemotherapy. PMID- 16039011 TI - Comment on "the beneficial effects of specialist thoracic surgery on the resection rate for non-small-cell lung cancer" by A.E. Martin-Ucar, D.A. Waller, J.L. Atkins, D. Swinson, K.J. O'Byrne, M.D. Peake [Lung Cancer 46 (2) (2004) 227 232.]. PMID- 16039012 TI - Unidirectional gas flow in soil porosity resulting from barometric pressure cycles. AB - During numerical simulation of air flow in the vadose zone, it was noticed that a small sinusoidal pressure would cause a gradual one-way migration of the pore gas. This was found to be a physical phenomenon, not a numerical artifact of the finite element simulation. The one-way migration occurs because the atmospheric pressure, and hence the air density, is slightly greater when air is flowing into the ground than when air is flowing out of the ground. A simple analytic theory of the phenomenon is presented, together with analytic calculations using actual barometric pressure data. In soil of one Darcy permeability, the one-way migration is of the order of a few tenths of a meter per year for either plane flow from ground surface or for radial flow from an open borehole. The migration is sufficiently small that it will have no practical consequences in most circumstances; however, investigators who conduct detailed numerical modeling should recognize that this phenomenon is not a numerical artifact in an apparently linear system. PMID- 16039013 TI - Validation of surgical site infection surveillance data in Scotland. AB - Validation of surveillance data is necessary to ensure its scientific credibility, to identify methodological problems within the surveillance programme, to help increase compliance and participation in the surveillance programme, and to identify data quality issues at local level. Surgical site infection surveillance (SSIS) in Scotland has been implemented in collaboration between Health Protection Scotland (HPS) and staff in acute divisions in Scotland. A team at HPS carried out a study to validate the SSIS data reported to them. The aims of the validation study were: (i) to measure the completeness of the denominator data; (ii) to measure the accuracy of all SSIS data items reported to HPS; and (iii) to determine the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the SSIs reported to HPS against the SSIs validated as part of this study. The methodology utilized for validation of SSIS data was based on an evaluation research approach. The evaluation research approach involves a range of investigative activities, aimed at judging the worth of a programme or practice, and measures SSIS in terms of structure, process and outcome. The completeness of the denominator and the means of identifying eligible patients was identified. Descriptive information about how SSIS data were collected and managed at hospital level was collated, and the accuracy and completeness of the reported SSIS data were measured by case note review of selected cases. SSIS data from 27 hospitals in 15 acute divisions and one special health board were validated. The results indicated that a total of 91% of the procedures carried out (denominator) during a specified three-month period were reported to HPS. The case notes validated over 90% of records reported to HPS; however, there was variation in data quality between hospitals. The sensitivity, specificity, PPV and NPV of the SSIs reported to HPS were 96.7, 99.0, 94.6 and 99.4%, respectively. Where problems with data were identified at local level, hospitals have been offered guidance to improve their data. As a result of this study, HPS are confident that the Scottish SSIS data are reliable and robust. PMID- 16039014 TI - Peri-operative glucose control and development of surgical wound infections in patients undergoing coronary artery bypass graft. AB - Elevated blood glucose following coronary artery bypass graft (CABG) is associated with an increased risk of surgical wound infection (SWI). It is unclear whether hyperglycaemia, the diabetic state, the longstanding vascular effects of diabetes, or the systematic inflammatory response confers the increased vulnerability to SWI. This study was designed to examine the significance of postoperative blood glucose control as a risk factor for SWI after vein graft harvesting on the leg and sternotomy. Patients with and without diabetes had a CABG within 60 days to be eligible. The present study was part of a larger protocol investigating SWI following CABG in a total of 374 patients. Potential risk factors, duration of diabetes, pre-operative glycated haemoglobin (HbA(1c)) and presence of long-term complications were recorded. All patient records were reviewed retrospectively to record 10% glucose infusions during the operation, and blood glucose concentrations and insulin therapy on postoperative days 0, 1 and 2. Patients were contacted by telephone 30 and 60 days after surgery and interviewed in accordance with a questionnaire about symptoms and signs of wound infection. In the present study, it was not possible to separate the effect of diabetes as a risk factor for SWI from that of hyperglycaemia. However, in the subgroup of patients without a pre-operative diagnosis of diabetes, increased blood glucose concentrations during postoperative days 0, 1 and 2 was associated with an increased risk of mediastinitis. PMID- 16039015 TI - Prognostic factors for nosocomial bacteraemia outcome: a prospective study in a Greek teaching hospital. AB - Epidemiological and clinical features were studied as potential prognostic factors for outcomes of bacteraemic patients in a tertiary care teaching hospital in Greece. The prospective study was conducted over 12 months and enrolled 153 consecutive hospital-acquired bacteraemic episodes in 137 patients. The crude in hospital mortality rate was 27% (37/137). The mean lengths of stay and of antimicrobial treatment were significantly longer for patients with a fatal outcome than for survivors (P<0.0001 and P=0.001, respectively). Needs for mechanical ventilation, urinary catheters and parenteral nutrition before or during the onset of episodes were significantly associated with fatalities [odds ratio (OR)=5.54, 95% confidence intervals (CI) 2.22-14.0, P<0.0001; OR=0.70, 95% CI 0.63-0.79, P=0.021; OR=5.03, 95% CI 1.88-13.95, P<0.0001, respectively]. Also, in logistic regression analysis, mechanical ventilation (OR=5.5, 95% CI 1.2-7.9, P=0.02) and parenteral nutrition (OR=8.8, 95% CI 3.8-11.4, P=0.003) were independent predictors of mortality. No differences between fatalities and survivors were found in sex, age, smoking habit, intensive care unit hospitalization, need for a nasogastric catheter and previous surgery. Neutropenia was associated with a fatal outcome (OR=3.65, 95% CI 1.24-10.91, P=0.006). None of the bacterial pathogens were significantly associated with an adverse outcome, whereas Staphylococcus aureus was recovered more frequently from survivors (P=0.02). Fatalities were more often associated with an intravascular catheter origin (P=0.002), whereas bacteraemias in survivors were associated with a skin/soft tissue origin (P=0.02). Various prognostic factors were associated with outcome in our bacteraemic population, and can be employed to identify bacteraemic patients at risk of death and to develop local strategies for its prevention. PMID- 16039016 TI - Enhanced multisensory integration in older adults. AB - Information from the different senses is seamlessly integrated by the brain in order to modify our behaviors and enrich our perceptions. It is only through the appropriate binding and integration of information from the different senses that a meaningful and accurate perceptual gestalt can be generated. Although a great deal is known about how such cross-modal interactions influence behavior and perception in the adult, there is little knowledge as to the impact of aging on these multisensory processes. In the current study, we examined the speed of discrimination responses of aged and young individuals to the presentation of visual, auditory or combined visual-auditory stimuli. Although the presentation of multisensory stimuli speeded response times in both groups, the performance gain was significantly greater in the aged. Most strikingly, multisensory stimuli restored response times in the aged to those seen in young subjects to the faster of the two unisensory stimuli (i.e., visual). The current results suggest that despite the decline in sensory processing that accompanies aging, the use of multiple sensory channels may represent an effective compensatory strategy to overcome these unisensory deficits. PMID- 16039017 TI - [Are anti-TNFAb indicated in systemic necrotizing vasculitides?]. PMID- 16039018 TI - Energy requirement in COPD. PMID- 16039020 TI - Uptake of varicella vaccine--a cross sectional survey of parental attitudes to nationally recommended but unfunded varicella immunisation. AB - The aim of this study was to assess the uptake of varicella vaccine in South Australian children under circumstances where varicella immunisation is recommended, but is not funded by Government. The study examined the main reasons that determined a parent's decision whether or not to have their child immunised with varicella vaccine. A cross-sectional survey was conducted by Computer Aided Telephone Interviews (CATI) in June 2004. Data were obtained from 613 households containing 1148 children aged from birth to 17 years of age. Statistical analyses were performed using data weighted to the South Australian population. Six hundred and eighty children (55.7%) had a history of varicella infection and 446 children (42.0%) had received varicella vaccine (weighted data). The most common reasons cited for not having children immunised included lack of knowledge about the vaccine and cost. One year after inclusion of varicella vaccine in the Australian Standard Vaccination Schedule there is evidence of incomplete coverage in children in South Australia due to absence of government funding for vaccine provision. PMID- 16039021 TI - Investigation of non-typeable Haemophilus influenzae outer membrane protein P6 as a new carrier for lipooligosaccharide conjugate vaccines. AB - Non-typeable Haemophilus influenzae (NTHi) outer membrane protein P6 was used as a new protein carrier for NTHi detoxified lipooligosaccharide (dLOS) conjugates due to its conservation and potential to elicit bactericidal antibodies. P6 was covalently conjugated to dLOS of strain 9274 through adipic acid dihydrazide with different ratios of dLOS to P6, which resulted in two conjugate formulations with weight ratios of dLOS to P6 of 3.7 for dLOS-P6 (I) and 1.6 for dLOS-P6 (II). Binding activity of the conjugates was examined by an enzyme-linked immunosorbent assay with mouse monoclonal antibodies specific to LOS and P6 and a rabbit anti P6 serum. The results showed that the conjugates bound not only to the LOS antibody but also to both P6 antibodies, suggesting that the conjugates retained epitopes of both LOS and P6 antigens. Animal studies revealed that dLOS-P6 (II) induced high levels of anti-LOS and anti-P6 IgGs in mice and rabbits. However, dLOS-P6 (I) induced lower levels of anti-LOS IgGs in mice and rabbits and anti-P6 IgGs in rabbits with no anti-P6 IgGs in mice. In addition, all rabbit, but not mouse, antisera elicited by the conjugates showed bactericidal activity against the homologous strain, and two of them elicited by each conjugate plus Ribi adjuvant showed cross-bactericidal activity against three of five major serotype stains. These data indicate that P6 could serve as an effective carrier for dLOS or other carbohydrate conjugates and that the ratio of carbohydrate to P6 might contribute to immune responses in vivo. PMID- 16039022 TI - Contribution of muscular weakness to osteoporosis: computational and animal models. AB - BACKGROUND: Chronic weakness of the femoral musculature with old age may result in prolonged exposure of bone to critical understressing and, thus, cause osteoporotic changes. This study aims at quantifying long-term changes in thickness and mechanical properties of trabecular bone at the proximal femur due to muscular weakness. METHODS: We utilized computational models of typical planar trabecular lattices at the proximal femur for simulating long-term changes in morphological and mechanical properties of trabecular bone. Incorporating cellular communication network with osteocytes as mechanosensors, the models were able to mimic mechanotransduction and consequent thickening and/or thinning of individual trabeculae in response to altered gluteus muscle and hip joint loads. We also studied a rat model (n=14) in which we surgically detached the gluteus muscle, to approximately 50% or completely. FINDINGS: The computational simulations showed that when the force of the gluteus decreased (with or without simultaneous decrease in hip joint load), the most dramatic degradation in bone density, strength and stiffness occurred at the greater trochanter. Animal studies also demonstrated significant thinning of femoral trabeculae after 19 weeks of adaptation. Specifically, Tukey-Kramer analysis showed that rats subjected to partial surgical detachment of the gluteus had femoral trabeculae that were 22% thinner than controls (P<0.05). INTERPRETATION: The present study showed that in both the computer and animal models, manipulation of muscle loading produced a significant stimulus for bone to adapt, i.e., a stimulus that is beyond its irresponsive 'lazy zone'. Accordingly, the results obtained herein indicate that muscular weakness may be an important factor contributing to osteoporosis. PMID- 16039023 TI - Interactions of mycorrhizal fungi with Pteris vittata (As hyperaccumulator) in As contaminated soils. AB - A greenhouse trial was conducted to investigate the role of arbuscular mycorrhizas (AM) in aiding arsenic (As) uptake and tolerance by Pteris vittata (As hyperaccumulator) and Cynodon dactylon (a multi-metal root accumulator). Plants inoculated with lived and killed native mycorrhizas isolated from an As mine site were grown in a sterile and slightly acidic soil. The infectious percentage of mycorrhizas (0 mg/kg As: 26.4%, 50 mg/kg As: 30.3%, 100 mg/kg As: 40.6%) and the average biomass of shoots in infected P. vittata increased (0 mg/kg As: 2.45 g/pot, 50 mg/kg As: 2.48 g/pot, 100 mg/kg As: 10.9 g/pot) according to the increase of As levels when compared to control. The indigenous mycorrhizas enhanced As accumulation (0 mg/kg As: 3.70 mg/kg, 50 mg/kg As: 58.3 mg/kg; 100 mg/kg As: 88.1 mg/kg) in the As mine populations of P. vittata and also sustained its growth by aiding P absorption. For C. dactylon, As was mainly accumulated in mycorrhizal roots and translocation to shoots was inhibited. PMID- 16039024 TI - Young teenagers' perceptions of their own and others' bodies: a qualitative study of obese, overweight and 'normal' weight young people in Scotland. AB - The rise in numbers of overweight/obese children in the UK is causing widespread concern. Biomedical constructions of body acceptability and 'good health' mean that overweight/obese young people are frequently seen as deviant. The socio cultural contexts within which young teenagers become fat, and lay conceptualisations of fatness, have largely been ignored. This qualitative study involved in-depth interviews with teenagers aged 13-14 years (n = 36), drawn from families living in areas classified as socio-economically disadvantaged. Half of the sample had a Body Mass Index (BMI) classifying them as overweight or obese, whilst the remainder were classified as being 'normal' weight. Participants' embodied perceptions of fatness were complex and sometimes contradictory. We discuss what young teenagers perceive the influences on fatness and body size to be; the professed consequences of being fat; participants' experiences of attempting to lose weight; and, their reported interactions with friends and family relating to fatness and dieting. Participants rarely mentioned any health related consequences of their own and others' fatness, although wearing 'nice' clothes and being slowed down were raised as considerations by girls and boys, respectively. 'Normal' weight teenagers who disliked their bodies or who wanted to lose weight often claimed to be anxious about this. Being very obese also led to anxiety and reported attempts at 'crash dieting'. Acceptance of body size/shape was, however, common amongst the overweight and obese teenagers, although some had attempted weight loss. The teenagers in this study were rarely supportive of friends or family who attempted to lose weight and frequently disagreed with others' perceptions of fatness. These findings are important as they contradict the common perception that being overweight/obese is related to body dissatisfaction and that young people have a fear of fatness. PMID- 16039025 TI - The healthy migrant theory: variations in pregnancy outcomes among US-born migrants. AB - Various studies have observed that infants born to foreign-born women have better birth outcomes (lower rates of preterm, low birth weight, and infant mortality) than those delivered to US-born women. While much attention has been given to the "healthy migrant effect" as an explanation for these positive outcomes, this theory has not been examined in an internally migrant population. The purpose of this study is to examine the relationship between maternal mobility history and birth outcomes among infants born to US resident mothers of Mexican origin. The study used 1995-1999 National Center for Health Statistics (NCHS) live birth/infant death cohort files of singleton infants delivered in the US to white women of Mexican origin (n = 2,446,253). Maternal mobility history (MMH), which refers to the relationship between the maternal place of birth and the state of residence at delivery, was categorized into the four following groups: (a) foreign-born-place of birth outside the US and delivery in the US; (b) outside region-place of birth in one US region and delivery in another US region; (c) within-region-place of birth in one US region and delivery in a different state in the same US region; and (d) within-state-place of birth and delivery in the same US state. Consistently, there is evidence to support the healthy migrant effect in an internally migrant population. Unique to this study are the findings that infants born to mothers with outside-region MMH had a lower risk of low birth weight (LBW) and small-for-gestational age (SGA) compared to those who did not move. Overall, this study provides evidence that the healthy migrant effect and its relationship to birth outcomes can be applied to an internally migrant population. PMID- 16039026 TI - Explaining educational-related inequalities in health: Mediation and moderator models. AB - This paper studies how education and certain lifestyle factors affect people's self-reported health. In addition to the assessment of the effects of education and lifestyle, the study contrasts two models of explaining educational-related health inequalities: the mediation model and the moderator model. The mediation model posits that well-educated people's better health, as compared to the poorly educated, is caused by their more 'healthy' lifestyles. The moderator model suggests, by contrast, that the effects of the lifestyle variables on health are dependent upon educational level. Several analyses are carried out on two large data sets comprising of middle-aged men and women in two Norwegian counties, Rogaland and Nordland. Two main findings are presented: (1) Both education and lifestyle factors have the expected effects on health. (2) The results do not permit a clear-cut conclusion as to which of the two models of educational related health inequalities should be preferred: whereas the results support the mediation model in the data from Rogaland, the moderator model is partially supported in the Nordland data. PMID- 16039027 TI - The voices of older women in a disadvantaged community: issues of health and social capital. AB - The voices of older women are rarely heard in debates about the health of disadvantaged groups. Despite growing interest in health in old age, the health experiences of older women as gendered social beings have yet to be fully explored. Their potential to contribute positively to family and community health is seldom acknowledged. The aim of this article is to present findings from a qualitative British Health Development Agency funded project on the relationship between social capital, health and gender, focusing on the health and social networks of older women in a socially disadvantaged community in the north of England. Seventy-seven community members were interviewed, of these 19 were older women aged 55-78 years. Their accounts of ill health in the context of ageing were analysed to explore the intricate ways in which social capital was created, maintained and linked to health. Findings suggest that social constructions of motherhood and caring underpinned responsibility for their own and others' health. Their experiences of dealing with health matters, together with frequent health talk, gave the women confidence as lay health experts, enabling them to contest medical advice. Drawing on personal experiences of trust and reciprocity, they recognised the importance of social networking in alleviating the problems of loneliness and isolation. At stressful times in their lives they were able to draw on existing support networks and, in spite of occasional personal conflicts, some benefited from the empowering and health-enhancing role of formal and informal participation in community life. These findings indicate that older women can operate autonomously in health matters and can substantially influence the development of healthy communities, although this can sometimes be at a personal cost. PMID- 16039028 TI - Cross-cultural perspectives on research participation and informed consent. AB - This study examined Portuguese Canadian and Caribbean Canadian immigrants' perceptions of health research and informed consent procedures. Six focus groups (three in each cultural group) involving 42 participants and two individual interviews were conducted. The focus groups began with a general question about health research. This was followed by three short role-plays between the moderator and the assistant. The role-plays involved a fictional health research study in which a patient is approached for recruitment, is read a consent form, and is asked to sign. The role-plays stopped at key moments at which time focus group participants were asked questions about their understanding and their perceptions. Focus group transcripts were coded in QSR NUDIST software using open coding and then compared across cultural groups. Six overriding themes emerged: two were common in both the Portuguese and Caribbean transcripts, one emphasized the importance of trust and mistrust, and the other highlighted the need and desire for more information about health research. However, these themes were expressed somewhat differently in the two groups. In addition, there were four overriding themes that were specific to only one cultural group. In the Portuguese groups, there was an overwhelming positive regard for the research process and an emphasis on verbal as opposed to written information. The Caribbean participants qualified their participation in research studies and repeatedly raised images of invasive research. PMID- 16039029 TI - The incubation of a social movement? Preterm babies, parent activists, and neonatal productions in the US context. AB - This article explores health-based activism on the part of the US 'parents of preemies' movement, a mutual-help network mobilized around babies born precariously early and acutely dependent upon life-support incubators. The movement articulates two meta-agendas for parental empowerment: (1) the quest to access/exercise greater participatory inclusivity vis-a-vis the preterm baby within the biomedical domain; and, (2) the quest to secure/command greater representational authority over the preterm baby within the public domain. Seen in terms of the erosion of the status quo, it can be argued that the movement's tangible and intangible aims to chip away at these traditions have been softly revolutionary: heralding new working partnerships between medical practitioners and patients' families; radical shifts in the technological consciousness and competences of preemie parents; and cyborg changes in conventional categories of the person. Yet, seen in terms of a normative order of things, it can be argued that the movement has largely and willingly been "co/operated": meaning that it has been "cooperative," but equally "co-opted" and "operated into" the disciplinary trajectory of neonatal medicine as well as the historical march of biopolitics with its governance of the collective body populous. From this critical perspective, the movement qua social movement thus itself might be considered incubated--cocooned, gestated, disciplined--and brought into existence by the very powers and hegemonic (patriarchal) machinery that viable resistance might struggle to govern instead of serve. PMID- 16039030 TI - The field of psychiatric contention in the UK, 1960-2000. AB - In this paper I discuss the question of how we should understand the concept of "social movements", particularly as applied to health related movements. My argument is that movements should be understood as "fields of contention". This concept, as I develop it, emphasizes two key aspects of social movement mobilization. Firstly, departing from traditional models of movements, which tend to view them as unified "things", it draws our attention to the numerous groups and agents who interact within the internal space of a "movement" and to the relations, alliances and conflicts between those various groups/agents as they unfold through time. Secondly, it draws our attention to the embedding of social movement struggles within multiple differentiated contexts of struggle, each of which affords different opportunities for struggle but each of which makes different demands upon activists if struggle is to prove effective. The model of fields of contention is explored within the paper using empirical data on a variety of "social movement organizations" (SMOs) which have formed around the mental health system in the UK over the last forty years. PMID- 16039031 TI - "To me, it's my life": medical communication, trust, and activism in cyberspace. AB - This paper studies the conversations and activities of an online support group for breast cancer sufferers and survivors and their supporters. Using communications medium theory and social capital theory, it examines the mundane and profound exchanges, the poignant self-disclosures, the creative expressions of solidarity, and the minor but not-insignificant political actions of people- initially strangers--who come together as a 'virtuous circle,' not only to assist with medical issues but also to meet emotional and even material needs. Sponsored by the Canadian nonprofit organization Breast Cancer Action Nova Scotia (BCANS), this virtual community has logged over a half million messages since 1996. Not every BCANS participant is an activist--many are just trying to grapple with their disease--but some find ways to shatter the professional "information monopoly," and to press for healthcare improvements. The study illustrates the scope, passion, and complexity of peer-to-peer medical communication in a virtual environment that promotes "thick trust". BCANS participants discuss with candor, warmth and even humor such painful topics as death and dying and the crises in intimate relationships brought about by a terminal illness. The sharing of confidences and fears enables participants to pool their 'collective intelligence' about many things, from how to cope with swelling, to how to think about end-of-life issues, to how to improve social policy. PMID- 16039032 TI - Gut dysfunction and endoscopic lesions after out-of-hospital cardiac arrest. AB - BACKGROUND: Cardiac arrest induces severe mesenteric ischaemia. The objective of this study was to assess the frequency of gut dysfunction and endoscopic lesions following resuscitation after cardiac arrest, and to evaluate the potential value of gut endoscopy performance in these circumstances. METHODS: This is a retrospective data files survey of 3617 patients from the database in a medical intensive care unit. A systematic review of medical and endoscopic files was performed within this database, using a standardised chart. PATIENTS: One-hundred and thirty consecutive patients who survived up to 48 h were admitted to our unit after out-of-hospital cardiac arrest. Seventy-eight of these patients (60%) presented with early clinical signs of gut dysfunction and/or lesions. Thirty-six patients underwent gut endoscopies (26%) and were included in the survey. RESULTS: Endoscopic lesions were observed in all cases; in 15 cases, gastrointestinal haemorrhage requiring intervention was identified. The occurrence of haemorrhagic and/or necrotic lesions was found to be associated with an initial rhythm of asystole, higher SAPS II values and epinephrine requirements, compared with cardiac arrest patients without such lesions. CONCLUSION: The frequent occurrence of endoscopic lesions in the presence of gut dysfunction following a cardiac arrest could suggest systematic gut endoscopy in such patients. However, an alternative recommendation would be to watch these patients very closely, treat all with prophylactic proton pump inhibitors, and endoscope only those with evidence of bleeding. PMID- 16039033 TI - The first year after successful cardiac resuscitation: function, activity, participation and quality of life. AB - AIM: To assess cognitive function, activities of daily living (ADL) and living situation longitudinally up to 1 year after cardiac arrest. SETTING: University hospital. MATERIALS: The study continued for 3 years and consecutive patients (18 75 years of age) who survived resuscitation were included. METHOD: A longitudinal study with examinations using the National Institute of Health Stroke Scale for assessing brain damage, along with the Mini Mental State Examination. To assess ADL, we used the Functional Independence Measure and the Instrumental Activity Measure. Life satisfaction was assessed, together with health-related quality of life. Social status and vocational status were recorded. RESULTS: Mortality was high during the first 90 days (31%). After discharge, not much improvement was seen in cognitive function. This was reflected in reduced dependency in ADL. Work capacity at 1 year was only 13%. The health related quality of life showed great improvement during the first year, being almost the same as for Swedish reference values. CONCLUSIONS: Most of the improvement resulting in the independence of ADL occurred during the first 45 days. It is important to give the next of kin information about the patient's progress and need for assistance in order to enable them to plan for the future before discharge. PMID- 16039034 TI - Relative effectiveness of interposed abdominal compression CPR: sensitivity analysis and recommended compression rates. AB - Interposed abdominal compression, IAC-CPR incorporates alternating chest and abdominal compressions to generate enhanced artificial circulation during cardiac arrest. The technique has been generally successful in improving blood flow and survival compared to standard CPR; however, some questions remain. OBJECTIVE: To determine "why does IAC-CPR produce more apparent benefit in some subjects than in others?" and "what is the proper compression rate, given that there are actually two compressions (chest and abdomen) in each cycle?" METHOD: Computer models provide a means to search for subtle effects in complex systems. The present study employs a validated 12-compartment mathematical model of the human circulation to explore the effects upon systemic perfusion pressure of changes in 35 different variables, including vascular resistances, vascular compliances, and rescuer technique. CPR with and without IAC was modeled. RESULTS AND CONCLUSIONS: Computed results show that the effect of 100 mmHg abdominal compressions on systemic perfusion pressure is relatively constant (about 16 mmHg augmentation). However, the effect of chest compression depends strongly upon chest compression frequency and technique. When chest compression is less effective, as is often true in adults, the addition of IAC produces relatively dramatic augmentation (e.g. from 24 to 40 mmHg). When chest compression is more effective, the apparent augmentation with IAC is relatively less (e.g. from 60 to 76 mmHg). The optimal frequency for uninterrupted IAC-CPR is near 50 complete cycles/min with very little change in efficacy over 20-100 cycles/min. In theory, the modest increase in systemic perfusion pressure produced by IAC can make up in part for poor or ineffective chest compressions in CPR. IAC appears relatively less effective in circumstances when chest pump output is high. PMID- 16039035 TI - A history of diabetes is associated with an adverse outcome among patients admitted to hospital alive after an out-of-hospital cardiac arrest. AB - BACKGROUND: Factors of importance for the outcome among patients who are admitted to hospital alive after an out-of-hospital cardiac arrest are not well described in the literature. The importance of a history of diabetes for the outcome among these patients has not been reported in detail previously. This survey aims to describe the outcome among patients who were admitted to hospital after an out-of hospital cardiac arrest in relation to whether they had a history of diabetes. METHODS: All patients who were admitted to hospital alive after an out-of hospital cardiac arrest in the two city hospitals in the Municipality of Goteborg between 1980 and 2002 were included in the survey. RESULTS: In all, 1377 patients fulfilled the inclusion criteria and 187 (14%) of them had a history of diabetes. Patients with diabetes differed from those without diabetes by having a previous history of myocardial infarction, angina pectoris, hypertension and heart failure more frequently. Furthermore, they were older, with a mean age of 70 years versus 66 years for patients without diabetes. Among patients with diabetes, 25% were discharged alive, as compared with 37% of patients without diabetes (p=0.002). When adjusting for differences at baseline, the adjusted odds ratio for diabetic patients being discharged alive (versus no diabetes) was 0.57 (95% confidence interval, 0.39-0.80). CONCLUSION: Among patients admitted to hospital after an out-of-hospital cardiac arrest, 14% had a history of diabetes. These patients had a lower survival rate compared with those without diabetes, even after correcting for dissimilarities at baseline. It remains to be determined whether an early metabolic intervention in these patients will improve survival. PMID- 16039036 TI - Epinephrine and vasopressin during cardiopulmonary resuscitation. AB - Epinephrine (adrenaline) and vasopressin have been by far the most commonly studied vasopressors in experimental cardiac arrest. Despite animal experimental studies suggesting improved outcomes in experimental cardiac arrest, clinical trials of pressor agents have failed to show clear cut benefit from either vasopressin or epinephrine, although few, if any, trials compared pressor agents to a placebo. The action of vasopressors in the heart, particularly beta1 adrenergic stimulation, is associated with adverse cardiac effects including post resuscitation myocardial dysfunction, worsening ventricular arrhythmias, and increasing myocardial oxygen consumption. Alpha2-adrenergic agonists, in experimental studies, show great promise in improving outcomes in experimental cardiac arrest, but have not been studied in humans. The combination of epinephrine and vasopressin may be effective, but has been incompletely studied. Clinical trials of vasopressor agents, which minimize direct myocardial effects are needed. PMID- 16039037 TI - Effect of nitric oxide synthase modulation on resuscitation success in a swine ventricular fibrillation cardiac arrest model. AB - BACKGROUND: We have demonstrated previously that the nitric oxide synthase (NOS) inhibitor N(G)-nitro-L-arginine (L-NNA) decreases free radical generation and nitrosative injury via peroxynitrite formation after epicardial dc shocks. OBJECTIVE: Our purpose was to explore the effects of NOS inhibition and NOS donation on cardiopulmonary resuscitation (CPR) success after cardiac arrest of variable duration. We used the non-selective NOS inhibitor L-NNA and the selective neuronal NOS inhibitor ARR-17477, the NOS donor S-nitroso-N acetylpenicillamine (SNAP) and the vasodilator Enalaprilat, which lowers arterial pressure via a non-NO mechanism. METHODS: Part I: 17 pigs undergoing 4 min supported (i.e. with closed-chest compression and ventilation) ventricular fibrillation (VF) were divided into two groups: a no-L-NNA group (n=8) receiving IV saline and an L-NNA group (n=9) receiving IV L-NNA (5 mg/kg) for 8 min before VF was induced. Part II: 35 pigs undergoing 6-8 min VF were randomized to three groups: a no-L-NNA group (n=13) receiving IV saline, an L-NNA group (n=11) receiving IV L-NNA (5 mg/kg) and an ARR17477 group (n=11) receiving IV ARR17477 (5 mg/kg) before VF. All animals in Part II underwent unsupported VF (no chest compression or ventilation) for 6 min (n=13) or 8 min (n=22); closed-chest compression, ventilation and epinephrine (adrenaline) were employed after defibrillation. Part III: 12 swine were divided into two groups: control (n=6) receiving saline and an LNNA group (n=6) receiving IV LNNA (5 mg/kg). Swine underwent 6 min unsupported VF and 2 min supported VF before defibrillation. Part IV: 25 animals were studied to determine the effect of the NO donor SNAP and the angiotensin-converting enzyme inhibitor Enalaprilat on coronary perfusion pressure (CPP). RESULTS: In Part I, after defibrillation, with continued ventilation, chest compression and epinephrine, 8/9 L-NNA pigs achieved ROSC versus 4/8 control pigs (p=0.11). After 60 s of CPR, 7/9 pigs in the L-NNA group achieved ROSC versus 2/8 pigs in the no-L-NNA group (p<0.05). Only 2/9 pigs receiving L-NNA required epinephrine (1 mg) after defibrillation, compared to 6/8 pigs requiring at least one dose of epinephrine in the no-L-NNA group (p<0.05). In Part II, there was no significant difference between L-NNA, ARR17477 and control pigs in ROSC. However, control pigs required 6.8+/-1.4S.E. mg epinephrine; L-NNA pigs and ARR17477 pigs required less epinephrine (3.7+/-0.7 and 3.0+/-0.3 mg, both p=0.01). Shorter chest compression was required in the L NNA group (252+/-38 s, p<0.05) and in ARR17477 group (222+/-15 s, p<0.05) compared to the control group (405+/-77 s). In Part III, L-NNA infusion caused a significant increase in mean blood pressure at baseline, but did not change CPP throughout the experiment. In Part IV, there were no significant differences in the changes of mean blood pressure and CPP between SNAP and Enalaprilat group in all animals throughout the experiment. CONCLUSION: NOS inhibition pre-arrest did not improve survival, but did reduce requirements for epinephrine and closed chest compression in a swine resuscitation model. PMID- 16039038 TI - Structural and dynamic effects of single 7-hydro-8-oxoguanine bases located in a frameshift target DNA sequence. AB - DNA 7-hydro-8-oxoguanine (8-oxoG) is implicated in frameshift formation in an G(6) sequence of the HPRT gene in mismatch repair (MMR) defective cells. Using oligonucleotides based on this frameshift hotspot, we investigated how a single 8 oxoG modified the structural and dynamic properties of the G(6) tract. A 30 ns molecular dynamics (MD) simulation indicated compression of the minor groove in the immediate vicinity of the lesion. Fluorescence polarization anisotropy (FPA) and MD demonstrated that 8-oxoG increases DNA torsional rigidity and also constrains the movement of the single-stranded region at the single/double stranded DNA junction of model DNA replication template/primer. These constraints influenced the efficiency of primer extension by Klenow (exo(-)) DNA polymerase. PMID- 16039039 TI - Activity coefficients and thermodynamic parameters for RbCl/CsCl + amide (acetamide, propanamide, and n-butanamide) + water system at 298.15 K. AB - Electromotive force (emf) of the chemical cell without liquid-junction K-ISE | RbCl/CsCl ( m(E)) | ISE-Cl and K-ISE | RbCl/CsCl (m(E)), amide (m(N)) | ISE-Cl, have been measured at 298.15 K, where m(E)=(0.005 to 0.5) mol kg(-1) and m(N)= (0.05 to 3.0) mol kg(-1). The activity coefficients of RbCl/CsCl in amide (acetamide, propanamide, and n-butanamide) + water mixture can be obtained from these electromotive force data and in the mean time the Gibbs free energy interaction parameters of RbCl/CsCl + amide pair in water, g(EN), as well as the salt constant, k(S), can be evaluated. The results show that both g(EN)>0, k(S)>0 at 298.15 K, and all the activity coefficients of electrolyte in amide + water mixture increase with increasing the m(N), but it is a little complicated for the dependence of activity coefficients on m(E). These thermodynamic parameters were discussed in terms of a model of the structural interaction and electrostatic interaction and the dependence of them on the number of carbon atoms in amide as well as the radius of metal ions were interpreted by the group additivity principle. PMID- 16039040 TI - MK-886 enhances tumour necrosis factor-alpha-induced differentiation and apoptosis. AB - We investigated the role of the 5-lipoxygenase (5-LOX) pathway of arachidonic acid metabolism in tumour necrosis factor-alpha (TNF-alpha)-induced differentiation of human leukemic HL-60 cells using MK-886, an inhibitor of 5-LOX activating protein. MK-886 augmented cell cycle arrest and differentiation induced by TNF-alpha; however, both effects were probably 5-LOX-independent, because a general LOX inhibitor, NDGA, had no effect. Apoptosis was significantly elevated after combined TNF-alpha and MK-886 treatment, which could be partially associated with changes of Mcl-1 protein expression. NF-kappaB signalling or activation of JNKs were not modulated by MK-886. Thus, in addition to apoptosis, MK-886 can enhance TNF-alpha-induced differentiation. PMID- 16039041 TI - Radiotherapy for primary thyroid cancer as a risk factor for second primary cancers. AB - Although radiation is considered a risk factor for thyroid cancer, the potential relationship between radiation therapy and the risk of second primary cancer among patients with first primary thyroid cancer has not been evaluated. We identified 26,639 patients with first primary thyroid cancer in the Surveillance, Epidemiology, and End Results (SEER) database from 1973 to 2000. Information on radiation therapy as well as second primary cancers was recorded in SEER. The proportional hazards model was utilized to estimate adjusted risk ratios (RRs) and their 95% confidence intervals (CIs) to assess the potential association between radiation therapy for thyroid cancer and the risk of second primary cancers. With 270,674.33 person-years of follow-up, 1,896 (7.1%) of the 26,639 patients with first primary thyroid cancer developed second primary cancers. Among the second primaries, 35 occurred in the thyroid. No obvious association was observed between radiation therapy and the overall risk of second primary cancer after ten years of follow-up (RR=1.07, 95% CI=0.88-1.30). However, an increased risk was seen for several cancers, including upper digestive system cancers (RR=1.66, 95% CI=1.07-2.57) and myeloid malignancies (RR=3.26, 95% CI=1.39-7.67). Radiation therapy was associated with reduced second cancer risks for thyroid cancer (RR=0.18, 95% CI=0.04-0.76). Beam radiation might be important to the digestive system, radioactive implants might be associated with the male genital system, radioisotopes might have an effect on myeloid malignancies, and combined beam radiation with radioactive implants or radioisotopes might be related to the increased risk of respiratory system cancers. This study suggests that radiation therapy for patients with first primary thyroid cancer might be associated with an increased risk of developing a second primary cancer in the upper digestive system and second primary myeloid malignancies. Radiation therapy for adult patients with thyroid cancer might be associated with a reduced risk of second primary thyroid cancer. PMID- 16039042 TI - In vitro antileukaemic activity of extracts from berry plant leaves against sensitive and multidrug resistant HL60 cells. AB - The aim of the present study was to determine in vitro antileukaemic activity of extracts obtained from selected berry plant leaves (Fragaria x ananassa Duch. cv Elsanta, raspberry Rubus ideus L. cv Polana and blueberry Vaccinium corymbosum L. cv Bluecrop) against promyelocytic HL60 cell line and its multidrug resistant sublines exhibiting two different MDR phenotypes: HL60/VINC (overexpressing P glycoprotein) and HL60/DOX (overexpressing MRP1 protein). It was found that the blueberry extract was the most efficient against sensitive HL60 cell line (about 2-fold more active than strawberry and raspberry extracts) but presented much lower activity towards resistant cells. In contrast, strawberry and raspberry extracts exhibited the high cytotoxic activity against sensitive leukaemia HL60 cell line as well as its MDR sublines. The values of resistance factor (RF) found for these extracts were very low lying in the range 0.32/2.0. PMID- 16039043 TI - Resection of lung metastasis from gallbladder carcinoma: immunohistochemistry of RCAS1 and CD8+T cells in primary and metastatic tumors. AB - Advanced Gallbladder cancer has an extremely poor prognosis. We examined a patient with resectable gallbladder cancer with associated lung metastasis. A 64 year-old female patient, diagnosed with gallbladder cancer and a solitary benign lung tumor by imaging, was subjected to extensive cholecystectomy and extrahepatic bile duct resection. After one year, a follow-up CT indicated enlargement of the lung tumor; video-assisted right middle lobectomy was then performed. The lung tumor was diagnosed as a metastasis derived from the gallbladder cancer by pathology and immunohistochemistry. Expression of RCAS1, an independent unfavorable prognostic indicator in gallbladder cancer, was observed in both the gallbladder and lung tumors. However, infiltration of CD8+T cells was only seen in the lung metastatic tumor. She has remained free of any evidence of recurrence in the 10 months and 4 years after the first surgery. The results that metastasis is solitary and infiltrated by CD8+T cells correspond with the present clinical history. PMID- 16039044 TI - Enhancing effect of iron on chromate reduction by Cellulomonas flavigena. AB - Cr(VI) is considerably toxic and the detoxification of Cr(VI) is of great importance. This study investigated the effect of iron on Cr(VI) reduction by Cellulomonas flavigena. The results demonstrated that addition of FeCl3 or lepidocrocite promoted Cr(VI) reduction, with the reduction ratio of above 90 and 80% achieved, respectively, but addition of hematite did not lead to the increase of reduction ratio, which suggests that the effect of iron on chromate reduction appears different with the diversity of iron-oxides. In this study, the effect of initial Cr(VI) and Fe(III) concentration on Cr(VI) reduction and the change of pH value were also investigated. The reduction ratio increased with the increase of the initial concentration ratio of Fe(III)/Cr(VI). The value of pH gradually increased from 7.0 to around 9.0. PMID- 16039045 TI - The development of the globally harmonized system (GHS) of classification and labelling of hazardous chemicals. AB - The hazards of chemicals can be classified using classification criteria that are based on physical, chemical and ecotoxicological endpoints. These criteria may be developed be iteratively, based on scientific or regulatory processes. A number of national and international schemes have been developed over the past 50 years, and some, such as the UN Dangerous Goods system or the EC system for hazardous substances, are in widespread use. However, the unnecessarily complicated multiplicity of existing hazard classifications created much unnecessary confusion at the user level, and a recommendation was made at the 1992 Rio Earth summit to develop a globally harmonized chemical hazard classification and compatible labelling system, including material safety data sheets and easily understandable symbols, that could be used for manufacture, transport, use and disposal of chemical substances. This became the globally harmonized system for the Classification and Labelling of Chemicals (GHS). The developmental phase of the GHS is largely complete. Consistent criteria for categorising chemicals according to their toxic, physical, chemical and ecological hazards are now available. Consistent hazard communication tools such as labelling and material safety data sheets are also close to finalisation. The next phase is implementation of the GHS. The Intergovernmental Forum for Chemical Safety recommends that all countries implement the GHS as soon as possible with a view to have the system fully operational by 2008. When the GHS is in place, the world will finally have one system for classification of chemical hazards. PMID- 16039046 TI - P-selectin glycoprotein ligand-1 variable number of tandem repeats (VNTR) polymorphism in patients with multiple sclerosis. AB - P-selectin glycoprotein ligand-1 (PSGL-1) is an important adhesion molecule involved in lymphocyte recruitment into the brain, which represents a crucial step in the pathogenesis of multiple sclerosis (MS). Three hundred twenty-one MS patients and 342 controls were genotyped for the presence of a polymorphism in the PSGL-1 gene, consisting of a variable number of tandem repeats (VNTR) originating three possible alleles: A, B and C, in order to test whether they influence the susceptibility and the course of the disease. No significant differences among allelic frequencies of A, B and C alleles in MS as compared with controls were observed. Stratifying patients according to the course of the disease, a significantly increased frequency of the shortest C allele in PP-MS was found (7.1%), either in comparison with controls (P=0.011) or with all other MS patients, who had acute inflammatory attacks at onset and an initial RR form (P=0.036). Besides, none of SP-MS patients was a carrier of the C allele and B carriers converted later from RR to SP course as compared with A/A subjects (after 15.8 rather than 8.8 years, P=0.01). In conclusion, the C allele of the VNTR polymorphism in PSGL-1 is likely to be associated with PP-MS. As this allele has been demonstrated to have a very low efficiency in mediating lymphocyte binding to brain endothelium during attacks, its high frequency in PP-MS could be related to the absence of exacerbations in such patients. PMID- 16039047 TI - Expression of aquaporin-1 immunoreactivity by photoreceptor cells in the mouse retina. AB - Aquaporin water channels play a crucial role in the maintenance of ionic and osmotic homeostasis in the neural tissue. In the sensory retina, aquaporin-4 is expressed by Muller glial cells, predominantly in the inner retina, while aquaporin-1 is expressed mainly in the outer retina. However, it is unknown whether aquaporin-1 expression occurs in Muller cells or photoreceptor cells. By using immunohistochemical staining of retinal slices from rds mice, we show that the immunoreactivity for aquaporin-1 disappears along with the photoreceptor cell degeneration. In suspensions of dissociated retinal cells from control mice, photoreceptor cells expressed aquaporin-1 immunoreactivity while Muller cells were largely devoid of staining. The data suggest that photoreceptor cells, but not Muller cells, express aquaporin-1 in the murine retina. PMID- 16039048 TI - Medial-lateral balance during stance phase of straight and circular walking of human subjects. AB - Right and left balance during human locomotion has been estimated by the distance between locations of the sequential right and left heel-strikes, or step width. During the stance phase of human locomotion one leg maintains medial-lateral balance for the progression. We focused our attention on this point, and medial lateral balance during straight-ahead and circular walking was investigated by recording sole-floor reaction force from five anatomically discrete points of human sole; calcaneus, 1st, 3rd, 5th metatarsals and great toe. Forces from these points were recorded during straight walking and circular walking. Medial-lateral balance was obtained by subtracting force at 5th metatarsal from force at 1st metatarsal (x-axis vector). The foot takes off the floor from medial balance in most steps, although in some steps the foot takes off from lateral balance at slower walking speed at 2 km/h or 4 km/h, showing variable patterns of x-axis vector. At faster walking speed at 6 km/h or 8 km/h body weight shifted to 1st metatarsal before taking off the floor. During circular walking body weight shift to 1st metatarsal in the outer foot, and to 5th metatarsal in the inner foot in most cases. PMID- 16039049 TI - Preference for caffeine appears earlier in non-anxious than in anxious mice. AB - Caffeine has reinforcing and anxiogenic properties. We have investigated whether the anxiety level of mice may influence their caffeine consumption. In a population of CD1 mice, animals were selected as anxious and non-anxious, using the elevated plus maze and the light/dark box tests. After a period during which we made mice able to associate the central effect of caffeine with the pipette delivering this substance, animals had free access to solvent (solution of sodium benzoate) and one of caffeine (0.3g/l) dissolved in this solvent. A progressive preference for the caffeine solution developed in both groups, but this occurred later in anxious than in non-anxious mice. Thus, it appears that the preference for caffeine is delayed in the anxious animals. PMID- 16039050 TI - Impaired exercise capacity, but unaltered mitochondrial respiration in skeletal or cardiac muscle of mice lacking cellular prion protein. AB - The studies of physiological roles for cellular prion protein (PrP(c)) have focused on possible functions of this protein in the CNS, where it is largely expressed. However, the observation that PrP(c) is expressed also in muscle tissue suggests that the physiological role of PrP(c) might not be limited to the central nervous system. In the present study, we investigated possible functions of PrP(c) in muscle using PrP(c) gene (Prnp) null mice (Prnp(0/0)). For this purpose, we submitted Prnp(0/0) animals to different protocols of exercise, and compared their performance to that of their respective wild-type controls. Prnp(0/0) mice showed an exercise-dependent impairment of locomotor activity. In searching for possible mechanisms associated with the impairment observed, we evaluated mitochondrial respiration (MR) in skeletal or cardiac muscle from these mice during resting or after different intensities of exercise. Baseline MR (states 3 and 4), respiratory control ratio (RCR) and mitochondrial membrane potential (DeltaPsi) were evaluated and were not different in skeletal or cardiac muscle tissue of Prnp(0/0) mice when compared with wild-type animals. We concluded that Prnp(0/0) mice show impairment of swimming capacity, perhaps reflecting impairment of muscular activity under more extreme exercise conditions. In spite of the mitochondrial abnormalities reported in Prnp(0/0) mice, our observation seems not to be related to MR. Our results indicate that further investigations should be conducted in order to improve our knowledge about the function of PrP(c) in muscle physiology and its possible role in several different neuromuscular pathologies. PMID- 16039051 TI - Human p53 tumor suppressor gene (TP53) and schizophrenia: case-control and family studies. AB - The human p53 tumor suppressor gene (TP53) is considered as a candidate susceptibility gene for schizophrenia because of its functions in neurodevelopment. To test for an association between TP53 and schizophrenia, both the case-control study and the transmission disequilibrium test (TDT) were performed on genotype data from eight polymorphisms in TP53. Our samples included 286 Toronto schizophrenia cases and 264 controls, and 163 Portuguese nuclear families. In the Toronto case-control study significant differences of allele frequencies of the CAA Ins/Del (p=0.027) and the 16bp Ins/Del (p=0.022) were detected. In TDT analysis we found significant differences for transmission of the CAA Ins/Del (p=0.017) in Portuguese schizophrenia families. Haplotype analysis also showed a significant association between TP53 and schizophrenia. These results provide further evidence that TP53 may play a role in the pathogenesis of schizophrenia. PMID- 16039052 TI - Seizures induced by microperfusion of glutamate and glycine in the hippocampus of rats pretreated with latrunculin A. AB - Changes in the membrane distribution of N-methyl-D-aspartate (NMDA) glutamate receptors seem to produce dramatic modifications in neuronal excitability and other properties of the neuron. In order to determine in vivo if these effects are due to the binding of extracellular glutamate and glycine to NMDA extrasynaptic receptors, we perfused the hippocampus of freely moving rats with the actin depolymerizant agent latrunculin A (4 microM) through microdialysis probes. One month later, continuous microperfusion of glutamate (1 mM) or glycine (1 mM) was used to induce epileptic seizures in the animals pretreated with latrunculin A. Glutamate microperfusion induced seizures in 50% of the animals studied, and glycine induced seizures in 75% of the rats. However, no effect was observed on control rats, or on those animals previously treated with picrotoxin. Simultaneous microperfusion of 100 microM MK-801 significantly reduced the number and duration of seizures induced by both glutamate and glycine. This study demonstrates that the application of latrunculin A results in long-term changes in susceptibility to the epileptogenic action of glutamate and glycine. PMID- 16039053 TI - Functional role of prostacyclin receptor in rat dorsal root ganglion neurons. AB - Recent studies on prostanoids showed that some of prostanoid receptors are expressed in rat dorsal root ganglion (DRG) neurons. These facts suggest that prostanoid receptors might be involved in the excitation mechanism of DRG neurons. In the present study, PCR experiments revealed that one of prostanoid receptor, prostacyclin receptor (IP receptor) was expressed in L6 and S1 rat DRG neurons and that the expression of IP receptor was not changed in DRG neurons obtained from the cyclophosphamide (CYP)-induced cystitis rat. We examined the functional role of IP receptor agonist and other prostanoids by measuring cyclic AMP (cAMP) accumulation and substance P (SP) release in primary cultured DRG neurons. The pretreatment of DRG neurons with prostanoid agonists such as iloprost (IP), butaprost (EP(2)), misoprostol (EP(2-4)), PGE(2) (EP(1-4)) or PGD(2) (DP and CRTH2) sensitized the DRG neurons and hence potentiated the lys bradykinin-induced SP release. The increase of SP release by lys-BK plus prostanoid agonists was proportion to cAMP accumulation. Iloprost was the most potent agonist to induce cAMP accumulation and SP release among prostanoid agonists evaluated in this study and its effect is mediated by IP receptor. Moreover, capsaicin-, ATP- and KCl-induced SP release was also enhanced by iloprost although iloprost did not change intracellular Ca(2+) and membrane depolarization induced by these chemical stimuli. These results strongly indicate that IP receptor play an important role in the sensitization of rat sensory neuron. PMID- 16039054 TI - Regulation of calcitonin gene-related peptide and TRPV1 in a rat model of osteoarthritis. AB - Pain in osteoarthritis (OA) remains an intractable problem in a majority of patients, with many of the commonly prescribed analgesics providing insufficient relief and considerable side effects. However, the structural or mechanistic cause of OA pain is still unknown. Animal models to address this issue have only recently been established, with much of the research to date focused on tissue pathology rather than pain. We have previously compared the surgically induced partial medial meniscectomy and chemically induced intra-articular iodoacetate injection rat models of OA in the rat, with reference to pain behaviour. This demonstrated relevant tissue pathology in both models, but greater evidence of pain related behaviour in the iodoacetate induced model. Here we further investigate the iodoacetate model using Fast Blue backlabelling from the articular joint space to identify the cell bodies of primary sensory afferents from the knee at the L4 dorsal root ganglion. Expression of calcitonin gene related peptide (CGRP) and the vanilloid receptor TRPV1 was quantified in these backlabelled cells and was enriched in the knee afferents in all animals studied, compared to the expression in neurons across the whole dorsal root ganglia (DRG). Analysis of the backlabelled population in the osteoarthritis model and controls showed an increase in both CGRP and TRPV1 expression in the iodoacetate model compared with control animals. Therefore, there is a potential role for CGRP and TRPV1 in the manifestation of pain behaviour accompanied by OA changes in the knee in the iodoacetate induced model. PMID- 16039055 TI - Association study between a functional glutathione S-transferase (GSTP1) gene polymorphism (Ile105Val) and tardive dyskinesia. AB - A possible role for oxidative stress in the pathophysiology of tardive dyskinesia (TD) has previously been proposed (reviewed in Andreassen and Jorgensen [O.A. Andreassen, H.A. Jorgensen, Neurotoxicity associated with neuroleptic-induced oral dyskinesias in rats Implications for tardive dyskinesia? Prog. Neurobiol. 61 (2000) 525-541]). Long-term administration of antipsychotics alters dopaminergic turnover, which results in increased formation of reactive oxygen species (ROS). This is hypothesized to lead to TD through neuronal toxicity as a consequence of oxidative stress. In the present study, the relationship between TD and a functional polymorphism of the gene coding for human glutathione S-transferase P1 (GSTP1), an important antioxidant enzyme involved in the detoxification of ROS, was studied in 225 chronic treatment-refractory patients with schizophrenia. An isoleucine (Ile) to valine (Val) substitution at codon 105 (Ile105Val) in the GSTP1 gene was genotyped. No significant difference in total AIMS scores was found among patients in the three genotype groups (chi(2)=1.47, d.f.=2, p=0.48). Moreover, no significant differences in genotype (chi(2)=0.05, d.f.=2, p=0.98) or allele frequencies (chi(2)=0.00, d.f.=1, p=1.00) were observed between subjects with and without TD. Our results suggest that the GSTP1 gene polymorphism does not confer increased susceptibility to TD, although further studies are warranted before a conclusion can be drawn. PMID- 16039056 TI - Early exposure to non-native language alters pre-attentive vowel discrimination. AB - The present study examined whether early exposure in language immersion would result in better pre-attentive discrimination of non-native speech sound contrasts. Mismatch negativity (MMN) responses were measured from two groups of Finnish children. The Monolingual group had no prior exposure to other languages than the native one, while the Immersion group consisted of children attending a French immersion program. The subjects were presented with two vowel contrasts in the oddball paradigm: the first pair was phonemic in the native language and the second was a within category pair in Finnish, but phonological in French. The results revealed that the Monolingual group showed a larger response to the native contrast in comparison with the non-native one, whereas both contrasts elicited a similar response in the Immersion group. These results suggest that early exposure to a new language enhances the pre-attentive discrimination ability reflected in increased MMN amplitude. PMID- 16039057 TI - Polymorphisms of myelin-associated glycoprotein gene are associated with schizophrenia in the Chinese Han population. AB - Results of gene expression microarray and quantitative PCR studies have suggested abnormalities in the expression of myelin-related genes including myelin associated glycoprotein (MAG) in schizophrenic patients. Research provides strong evidence for oligodendrocyte dysfunction in schizophrenics. In order to further assess the role of MAG in schizophrenia, we examined four single nucleotide polymorphisms (SNPs), namely rs2301600, rs3746248, rs720309 and rs720308, of this gene in Chinese schizophrenic patients (n=470) and healthy controls (n=470). The distribution of rs720309 T/A genotypes showed a strong association with schizophrenia (chi(2)=14.58, d.f.=2, P=0.0008). A haplotype constructed of rs720309-rs720308 also revealed a significant association with schizophrenia (chi(2)=11.914, d.f.=3, P=0.0084). Our findings of a significant associations between schizophrenia and the MAG gene suggest that this gene may be involved in susceptibility to schizophrenia in the Chinese Han population. PMID- 16039058 TI - High concentrations of plasma brain-derived neurotrophic factor in methamphetamine users. AB - Methamphetamine is a highly addictive drug that has a neurotoxic effect on the brain. A growing body of evidence suggests that brain-derived neurotrophic factor (BDNF) is associated with addictive behavior. The present study investigated the changes in plasma BDNF concentration that were induced by chronic methamphetamine use. Using an enzyme-linked immunosorbent assay (ELISA), we measured peripheral BDNF levels in methamphetamine users and in a control group. The plasma BDNF concentrations of methamphetamine users were significantly higher compared with those of controls (2536.3 pg/ml versus 1352.6 pg/ml). This finding suggests that BDNF plays some role in the neurotoxicity of methamphetamine. PMID- 16039059 TI - Serotonin metabolism in directly developing frog embryos during paternal care. AB - Central serotonin (5-HT) metabolism during embryogenesis and a 3-day post hatching period was analyzed using high performance liquid chromatography in the directly developing frog, Eleutherodactylus coqui. This anuran bypasses the free swimming larval stage and embryos hatch as miniature frogs in the adult phenotype. During embryogenesis and for a short time immediately after hatching, male E. coqui provide paternal care by brooding and guarding eggs/embryos to prevent desiccation and predation. Serotonin and its catabolite, 5-HIAA, were measured from whole brain during embryogenesis and at 3 days post-hatch to identify critical periods in 5-HT development and to determine the relationship between 5-HT and life history events such as hatching and frog dispersal from the nest site. Serotonergic activity was highest during the early-mid embryonic stages as indicated by the ratio of 5-HIAA/5-HT, a general indicator of turnover and metabolism. There were significant increases in tissue concentrations of 5-HT during the latest or terminal embryonic stage, just prior to hatching, and also at 3 days post-hatch, shortly before neonates disperse into the rainforest. These two increases probably represent different functional requirements during development. The first may occur as a result of the surge of development in the 5 HT system during late embryogenesis that occurs in E. coqui and the second may be from the increase demand in sensory and motor neural development required before dispersal from the nest site. PMID- 16039060 TI - Amburoside A, a glucoside from Amburanacearensis, protects mesencephalic cells against 6-hydroxydopamine-induced neurotoxicity. AB - This study evaluates the potential neuroprotective properties of amburoside A, a glucoside isolated from Amburana cearensis, on rat mesencephalic cell cultures exposure to the neurotoxin 6-hydroxydopamine (6-OHDA). The parameters determined were cell viability by the 3[4,5-dimethylthiazole-2-il]-2,5-diphenyltetrazolium bromide (MTT) method, nitric oxide (NO) and free radical formation by the measurement of nitrite concentration and thiobarbituric acid reacting substance (TBARS) formation as an indication of cellular lipid peroxidation. The results showed that AMB was less effective as a curative agent in the MTT assay, since its addition after 6-OHDA did not reverse the neurotoxin's effect, except at the highest concentration (AMB, 100 microg/ml). Similarly, the higher nitrite levels observed after exposure of the cells to 6-OHDA were only partially reversed by AMB, at this highest concentration. However, when AMB (0.5, 1, 10 and 100 microg/ml) was added before the toxin, it appeared to protect neuronal cells against 6-OHDA toxicity in a concentration-dependent manner, as shown by MTT assay. AMB also prevented free radical formation indicated by the increased nitrite concentration induced by 6-OHDA. Cells exposed to 6-OHDA showed a 3.4 times increase in TBARS concentration as compared to controls, and this effect was inhibited from 24% up to 64% by AMB (0.1-100 microg/ml), indicative of a neuroprotective effect. In conclusion, we show that AMB, acting as an antioxidant compound, presents a significant neuroprotective effect, suggesting that this compound could provide benefits as a therapeutic agent in neurodegenerative disease such as Parkinson's. PMID- 16039061 TI - Effect of nitration on protein tyrosine phosphatase and protein phosphatase activity in neuronal cell membranes of newborn piglets. AB - Protein tyrosine phosphatase predominantly determines the status of protein tyrosine kinase-dependent phosphorylation of specific proteins and controls the survival and death of neurons. Previous studies have shown that protein tyrosine phosphatase activity is decreased during hypoxia in cortical membranes of the newborn piglet. We have also shown that nitric oxide (NO) free radicals are generated during hypoxia, and may result in modification of protein tyrosine phosphatase via peroxynitrite-mediated modification. The present study tests the hypothesis that the hypoxia-induced decrease in protein tyrosine phosphatase activity is NO-mediated. To test this hypothesis, in vitro experiments were conducted by measuring protein tyrosine phosphatase activity in the presence of an NO donor, sodium nitroprusside (SNP), or peroxynitrite. Since 3-nitrotyrosine is produced as a consequence of peroxynitrite reactions, we have also examined the effect of 3-nitrotyrosine on protein phophatase activity. Cerebral cortical P(2) membranes were prepared from seven normoxic newborn piglets and each sample was divided into three aliquots: a control group, a SNP group (exposed to 200 microM SNP), and a peroxynitrite group (exposed to 100 microM peroxynitrite). Protein tyrosine phosphatase activity was determined spectrophotometrically in the presence or absence of 2 microM bpV(phen), a highly selective inhibitor of protein tyrosine phosphatase. The protein tyrosine phosphatase activity was 198+/ 25 nmol/mg protein/h in the normoxic group, 177+/-30 nmol/mg protein/h in the SNP group (p=NS versus normoxic) and 77+/-20 nmol/mg protein/h in the peroxynitrite group (p<0.001 versus normoxic). The results show that peroxynitrite but not SNP exposure results in decreased protein tyrosine phosphatase activity in vitro. Furthermore 3-nitrotyrosine (100 microm), a product of peroxynitrite, decreased the enzyme activity from 926+/-102 to 200+/-77 (p<0.001). We conclude that protein tyrosine phosphatase regulation is mediated by peroxynitrite. We propose that hypoxia-induced NO production leading to peroxynitrite formation is a potential mechanism of protein tyrosine phosphatase inactivation in vivo. The NO induced decrease in protein tyrosine phosphatase and protein phosphatase activity, leading to Bcl-2 protein phosphorylation and loss of its antiapoptotic activity may be a NO-mediated mechanism of programmed cell death in the hypoxic brain. PMID- 16039062 TI - Sensory stimulation reduces seizure severity but not afterdischarge duration of partial seizures kindled in the hippocampus at threshold intensities. AB - Epilepsy is a family of neurological disorders that result in seizure activity that is characterized by transient hypersynchronous activation of a large population of neurons. In animal models, focal tetanic electrical stimulation of sufficient duration and intensity, can elicit epileptiform activity, that if repeated results in progressive intensification of seizure activity known as kindling. Kindling serves as a model of partial as well as secondarily generalized temporal lobe epilepsy. We utilized hippocampal kindling to provide a means of evaluating the effect of sensory stimulation on the duration and severity of the induced seizure activity. Sensory stimuli targeted either the olfactory, auditory or somatosensory systems in an attempt to retard or suppress seizure activity. To that end, rats were chronically implanted with electrodes in the CA1 region of dorsal hippocampus and kindled once daily until the seizure behaviour was fully generalized. Kindling stimulation consisted of daily application of 1-s trains of biphasic square wave pulses applied at a frequency of 60Hz, at the afterdischarge (AD) threshold. Sensory stimulation was applied 6 8s after the kindling stimulation every third day. One group of rats received a different sensory stimulus (novel) every third day, while another group was presented with the same sensory stimulus (repeated) every third day. Kindling stimulation applied to the dorsal hippocampus resulted in progression of the AD characteristics and seizure behavior, which typically developed very slowly in the early stages. The application of both the novel and repeated sensory stimulation during partial seizures (stages 1 and 2) resulted in a reduction in the seizure severity but not in the afterdischarge duration. Sensory stimulation delivered during secondarily generalized seizures (stages 4 and 5) failed to affect either parameter. PMID- 16039063 TI - Comparison of protease-resistant prion protein inhibitors in cell cultures infected with two strains of mouse and sheep scrapie. AB - The transmissible spongiform encephalopathies (TSEs) are fatal neurodegenerative diseases. A primary therapeutic target for TSE intervention has been a protease resistant form of prion protein known as PrP(Sc) or PrP-res. In vitro testing of mouse scrapie-infected cell cultures has identified many PrP-res inhibitors that also have activity in vivo. Here we identify 32 new inhibitors of two strains of mouse scrapie PrP-res. Furthermore, to investigate the species-specificity of these and other PrP-res inhibitors, we have developed a high-throughput cell culture assay based on Rov9 cells chronically-infected with sheep scrapie. Of 32 inhibitors of murine PrP-res that were also tested in the Rov9 cells, only six showed inhibitory activity against sheep PrP-res. The three most potent inhibitors of both murine and ovine PrP-res formation (with 50% inhibition at < or =5 microM) were tannic acid, pentosan polysulfate and Fe(III) deuteroporphyrin 2,4-bisethyleneglycol. The latter two have anti-mouse scrapie activity in vivo. These results identify new inhibitors of murine and ovine PrP-res formation and reinforce the idea that compounds effective against PrP-res from one species or strain cannot be assumed to be active against others. PMID- 16039064 TI - Amino acids and transaminases activity in ventricular CSF and in brain of normal and Alzheimer patients. AB - The present study was conducted to determine the concentration of amino acids in the cerebrospinal spinal fluid (CSF) and the activities of two tramsaminases: glutamic oxaloacetate transaminase (GOT) and glutamic pyruvate transaminase (GPT) in human Alzheimer disease (AD) and normal brain. L-glutamic acid, L-glutamine and L-alanine are the most abundant amino acids in the CSF (50-55% of total amino acids). L-glutamine occurs at much higher levels in Alzheimer CSF compared to the normal CSF (229+/-91.8 nmol/ml in AD versus 107+/-47.2 nmol/ml in normal; P=0.0041). In contrast, L-aspartate occurs at significantly lower concentrations in Alzheimer CSF than normal CSF (46.1+/-25.7 nmol/ml in Alzheimer versus 95.2+/ 52.6 nmol/ml in normal; P=0.020). In Alzheimer brain (frontal, parietal and occipital cortices) GOT is present at significantly higher activities than in normal brain cortices (about 1.5 times higher; P<0.01). No significant differences for GPT activity occurred between normal and AD brain. Since CSF receives amino acids from brain tissues, and since GOT catalyzes the conversion of L-aspartate to L-glutamate, the higher concentrations of L-glutamine (which is derived from L-glutamate), and the lower concentrations of L-aspartate found in Alzheimer CSF could be considered as a consequence of the higher activity of GOT that occurs in Alzheimer brain. PMID- 16039065 TI - Genetic and biologic characteristics of Toxoplasma gondii infections in free range chickens from Austria. AB - The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in free-range chickens (Gallus domesticus) from 11 Bio-farms in Austria was determined. Antibodies to T. gondii assayed by the modified agglutination test (MAT) were found in 302 of 830 (36.3%) chickens with titers of 1:10 in 50, 1:20 in 69, 1:40 in 53, 1:80 in 40, 1:160 or higher in 90. Hearts of 218 chickens with MAT titers of 10 or higher were bioassayed individually in mice. Tissues from 1183 chickens were pooled and fed to 15, T. gondii-free cats. Feces of the cats were examined for oocysts; 11 cats shed T. gondii oocysts. T. gondii was isolated from 56 chickens by bioassay in mice. Thus, there were 67 isolates of T. gondii from these chickens. Genotyping of these 67 isolates using the SAG2 locus indicated that all 33 were Type II. Phenotypically and genetically these isolates were different from T. gondii isolates from Brazil. None of the isolates was virulent for mice. This is the first report of isolation of T. gondii from chickens from Austria. PMID- 16039066 TI - Inhibition kinetics of mushroom tyrosinase by copper-chelating ammonium tetrathiomolybdate. AB - With a strategy of chelating coppers at tyrosinase active site to detect an effective inhibitor, several copper-specific chelators were applied in this study. Ammonium tetrathiomolybdate (ATTM) among them, known as a drug for treating Wilson's disease, turned out to be a significant tyrosinase inhibitor. Treatment with ATTM on mushroom tyrosinase completely inactivated enzyme activity in a dose-dependent manner. Progress-of-substrate reaction kinetics using the two step kinetic pathway and dilution of the ATTM revealed that ATTM is a tight binding inhibitor and high dose of ATTM irreversibly inactivated tyrosinase. Progress-of-substrate reaction kinetics and activity restoration with a dilution of the ATTM indicated that the copper-chelating ATTM may bind slowly but reversibly to the active site without competition with substrate, and the enzyme ATTM complex subsequently undergoes reversible conformational change, leading to complete inactivation of the tyrosinase activity. Thus, inhibition by ATTM on tyrosinase could be categorized as complexing type of inhibition with a slow and reversible binding. Detailed analysis of inhibition kinetics provided IC50 at the steady-state and inhibitor binding constant (K(I)) for ATTM as 1.0+/-0.2 microM and 10.65 microM, respectively. Our results may provide useful information regarding effective inhibitor of tyrosinase as whitening agents in the cosmetic industry. PMID- 16039067 TI - Neoadjuvant chemotherapy with weekly paclitaxel in breast cancer patients may increase release of cancer cells into circulation by decreasing interstitial fluid pressure. PMID- 16039068 TI - Some experimental observations consistent with cell fusion hypothesis of cancer evolution. PMID- 16039069 TI - Salvage radiation therapy for residual superficial esophageal cancer after endoscopic mucosal resection. AB - PURPOSE: To analyze the outcomes of radiation therapy for patients with residual superficial esophageal cancer (rSEC) after endoscopic mucosal resection (EMR). METHODS AND MATERIALS: From May 1996 to October 2002, a total of 30 rSEC patients without lymph node metastasis received radiation therapy at Tohoku University Hospital and associated hospitals. The time interval from EMR to start of radiation therapy ranged from 9 to 73 days (median interval, 40 days). Radiation doses ranged from 60 Gy to 70 Gy (mean dose, 66 Gy). Chemotherapy was used in 9 of 30 patients (30%). RESULTS: The 2-year, 3-year, and 5-year overall survival rates and cause-specific survival rates were 91%, 82%, and 51%, respectively, and 95%, 85%, and 73%, respectively. The 2-year, 3-year, and 5-year local control rates for mucosal cancer were 91%, 91%, and 91%, respectively, and those for submucosal cancer were 89%, 89%, and 47%, respectively. These differences in survival rates for patients with two types of cancer were not statistically significant. Local recurrence and lymph node recurrence were more frequent in patients with submucosal cancer than in patients with mucosal cancer (p = 0.38 and p = 0.08, respectively). Esophageal stenosis that required balloon dilatation developed in 3 of the 30 patients, and radiation pneumonitis that required steroid therapy developed in 1 patient. CONCLUSIONS: Radiation therapy is useful for preventing local recurrence after incomplete EMR. PMID- 16039070 TI - Tumor vascularity evaluated by transrectal color Doppler US in predicting therapy outcome for low-lying rectal cancer. AB - PURPOSE: To evaluate the impact on T downstaging of the vasculature supplying blood flow to rectal cancer evaluated by color Doppler ultrasound. METHODS AND MATERIALS: Color Doppler images were graded in 29 T3-staged rectal carcinoma patients sonographically just before chemoradiation. Any arterial vessels detected in rectal masses were assigned one of two grades: vascularity was considered as grade 1 for vessels feeding the periphery and as grade 2 for vessels coursing in all rectal masses within its peripheral and central portions. The pulsatility indices (PI = peak systolic velocity - end-diastolic velocity/time-averaged maximum velocity) were calculated in the central and peripheral portions. RESULTS: The pathologic observations showed a change in stage in 15 of the 23 patients graded 2, positive predictive value 65.2% (p = 0.047), and in one of the six rectal cancers graded 1 (negative predictive value, 83.3%). The minimal peripheral PI values in rectal cancer graded 2 were higher in nonresponding (2.2 +/- 1.3) than in responding lesions (1.6 +/- 0.7) p = 0.01. CONCLUSION: Vascularity graded 2 associated with low peripheral PI values are indicators of therapy outcome. Vascularity graded 1 and high peripheral PI values in graded 2 have negative predictive value. PMID- 16039071 TI - Actin cytoskeleton disruption by ExoY and its effects on Pseudomonas aeruginosa invasion. AB - Three of the Type III-secreted effectors of Pseudomonas aeruginosa (ExoS, ExoT, and ExoY) each alter mammalian cell morphology in culture without causing a loss of cell viability. For ExoS and ExoT this property involves RhoGAP activity, and leads to actin cytoskeleton disruption and a reduced capacity for internalizing bacteria. ExoY does not possess RhoGAP activity. Instead, cell rounding depends upon its adenylate cyclase catalytic region. Since anti-phagocytic activities of ExoS and ExoT are associated with cell rounding and cytoskeleton disruption, we hypothesized that ExoY would also inhibit P. aeruginosa invasion of epithelial cells coinciding with adenylate cyclase-mediated cytoskeleton disruption. The results showed actin disruption of epithelial cells at 2 h post-infection associated with both adenylate cyclase-active ExoY and its catalytic mutant form ExoYK81M, and which coincided with inhibition of bacterial invasion (76% inhibition by ExoY, and 37% by ExoYK81M). Surprisingly, at 4h post-infection, neither form of ExoY inhibited invasion despite extensive actin disruption. These data suggest that ExoY, like ExoS and ExoT, contains more than one active domain affecting mammalian cell function. The data also suggest that cytoskeleton disruption does not necessarily predict invasion inhibitory activity, supporting the recently proposed model that P. aeruginosa internalization can proceed through more than one pathway. PMID- 16039072 TI - C. elegans homologue of the Caf1 gene, which encodes a subunit of the CCR4-NOT complex, is essential for embryonic and larval development and for meiotic progression. AB - The evolutionary conserved CCR4-NOT multi subunit complex is involved in different aspects of mRNA metabolism, including mRNA synthesis initiation and mRNA deadenylation (shortening of the poly(A) tail) in yeast and higher eukaryotes. Here we report the characterization of the gene encoding the Caf1 subunit of this complex in Caenorhabditis elegans, ccf-1, and the phenotypes associated with its inactivation. Use of staged populations and of mutants strains with altered germline showed that ccf-1 is predominantly expressed in embryos and adults. Loss of ccf-1 function, by both RNAi and a deletion allele, caused early embryonic and larval lethality. It also resulted in sterility in both males and hermaphrodites by blocking germ cell development at the pachytene stage of meiosis I. These results reveal that ccf-1 is an essential factor for both somatic and germline development in C. elegans. Functional analysis of ccf-1 may contribute to the understanding of the molecular role of the CCR4-NOT complex. PMID- 16039073 TI - Comparative analyses of sheep and human TRG joining regions: evolution of J genes in Bovidae is driven by sequence conservation in their promoters for germline transcription. AB - The availability of genomic clones representative of the T cell receptor gamma (TRG1@ and TRG2@) ovine loci enabled us to compare the germline genomic organization and nucleotide diversity of joining (J) segments and reconstruct their evolutionary history by phylogenetic analysis of cattle, sheep and human expressed sequences. Expression profiling (RT-PCR data) in fetus and adult indicated that only the ovine J genes in which two or more of the key sequence features, such as recombination signal sequences (RSS), 3' splice sites, and core sequences, are missing or severely altered fail to be transcribed. Comparative genomic examination of the two human with the six sheep germline transcription promoters located at 5' of the relevant constant (C)-distal J segments showed a strong conservation of the redundant STAT consensus motifs, indicating that TRG1@ and TRG2@ loci are under the influence of IL-7 and STAT signalling. These findings support the phylogenetic analysis of human and Bovidae (cattle and sheep) that revealed a different grouping pattern of C-distal compared to C proximal J segments. Likewise, the phylogenetic behaviour of either C-distal and C-proximal J segments is in accordance with the Bovidae TRG clusters evolution. Comparison of sheep and human structures of recombination signal sequences (RSS) has highlighted a greater conservation in sheep 12 RSS rather than 23 RSS thus suggesting that the initial recruitment of recombination activating genes (RAG) products requires at least one relatively high-affinity RSS per recombination event. PMID- 16039074 TI - An investigation of the variation in the transition bias among various animal mitochondrial DNA. AB - The transition:transversion ratio (ts/tv) is known to be very high in human mitochondrial DNA, but we have little information about this ratio in other species. Here we investigate the transition bias in animal mitochondrial DNA using single nucleotide polymorphism data at four-fold degenerate sites. We investigate this pattern of polymorphism in the cytochrome b gene (cyt-b) in 70 species using a total of 1823 mutations. We show that most species show a bias towards transitions but that the ratio varies significantly between species. There is little evidence for variation within orders or genera and between closely related species such as the great apes. The majority of the variation appears to be at a higher phylogenetic levels: between orders and classes. We test whether the variation in ts/tv ratio could be due to variation in the metabolic rate by considering whether the ratio is correlated to base composition. We find no evidence that the metabolic rate affects the ts/tv ratio. We also investigate the relative frequencies of C to T or T to C (C<-->T) mutations and A to G or G to A (A<-->G) mutations. We show that overall they occur at significantly different frequencies, and that there is significant variation in their relative frequency between species and between classes. We find no evidence in support of the hypothesis that this variation could be due to different metabolic rates. PMID- 16039075 TI - Heart rate responses to non-arousing trigeminal stimulation in infants: effects of sleep position, sleep state and postnatal age. AB - AIMS: The aim of this study was to examine the effects of maternal smoking, sleeping position, sleep state and postnatal age on heart rate changes following non-arousing trigeminal stimulation in infants. SUBJECTS: We studied healthy term infants, 13 of whom were born to mothers who did not smoke and 11 to mothers who smoked during pregnancy. Each infant was studied using daytime polysomnography on 3 occasions: (a) 2-3 weeks, (b) 2-3 months and (c) 5-6 months after birth. Nasal air-jet stimulation was presented in both active sleep (AS) and quiet sleep (QS) when infants slept both prone and supine. RESULTS: We found no difference between infants of smoking and non-smoking mothers in any of the parameters measured. Minimum HR (MinHR) following non-arousing trigeminal stimulation was significantly lower in the supine compared to the prone sleeping position at 2-3 weeks and 2-3 months of age (p<0.05) in AS, and at all 3 ages in QS (p<0.01). MinHR was significantly lower in QS compared to AS at 2-3 months when infants slept prone and at 5-6 months when sleeping supine (p<0.01). In QS, MinHR became lower with increasing postnatal age in both sleep positions (p<0.01). In AS, there was no maturational effect. The normalized bradycardia (DeltaHR%) was significantly greater in AS than in QS at 2-3 weeks of age (p<0.05) when infants slept supine. CONCLUSION: Our study has shown that there was a decrease in heart rate (MinHR) following trigeminal stimulation in infants up to 6 months of age and this was affected by sleep position and sleep state, being larger in the supine sleeping position and the QS state. PMID- 16039076 TI - Patterns of expression of steroidogenic enzymes during the first wave of the ovine estrous cycle as compared to the preovulatory follicle. AB - The expression patterns of steroidogenic enzymes in ovarian antral follicles at various stages of growth in a follicular wave have not been reported for sheep. Ovaries were collected from ewes (n=4-5 per group) when the largest follicle(s) of the first wave of the cycle, as determined by ultrasonography, reached (i) 3 mm, (ii) 4 mm, (iii) > or =5 mm in diameter or when there was a single (iv) preovulatory follicle in the last wave of the cycle, 12h after estrus detection. The expression pattern of steroidogenic enzymes was quantified using immunohistochemistry and grey-scale densitometry. The expression of CYP19 in the granulosa and 3beta-HSD and CYP17 in the theca increased (P<0.01) progressively from 3 to > or =5 mm follicles in the first wave of the cycle and was lower (P<0.01) in the preovulatory follicle compared to > or =5 mm follicles. However, the expression of 3beta-HSD in the granulosa increased (P<0.05) from 3 to > or =5 mm follicles and was maintained (P<0.05) at a high level in the preovulatory follicles. The amount of CYP19 in the granulosa of the growing follicles correlated positively (r=0.5; P<0.03) with the concurrent serum estradiol concentrations. We concluded that the expression pattern of steroidogenic enzymes in theca and granulosa of follicles growing in each wave in the ewe, paralleled with serum estradiol concentrations, with the exception that concentrations of 3beta-HSD in granulosa increased continuously from follicles 3mm in diameter to the preovulatory follicle. PMID- 16039077 TI - The effect of a steep high-frequency hearing loss on growth-of-masking functions in simultaneous masking for f(m)98%. In vivo assessment of [11C]1 was performed in a healthy Papio hamadryas baboon using positron emission tomography (PET). Following iv administration of [11C]1, significant accumulation was observed in the baboon brain and peripheral organs. In the brain, the radioactivity peaked at 20 min and remained constant for the duration of the imaging experiment. Pre treatment with the PBR-specific ligand, PK 11195 (5 mg/kg), effectively reduced the binding of [11C]1 at 60 min by 70% in the whole brain, whereas pre-treatment with the central benzodiazepine receptor ligand, flumazenil (1mg/kg), had no inhibitory effect on [11C]1 uptake. These results indicate that accumulation of [11C]1 in the baboon represents selective binding to the PBR. These exceptional in vivo binding properties suggest that [11C]1 may be useful for imaging the PBR in disease states. Furthermore, [11C]1 represents the first ligand of its pharmacological class to be labelled for PET studies and therefore has the potential to generate new information on the pathological role of the PBR in vivo. PMID- 16039132 TI - Highly antiproliferative, low-calcemic, side-chain ketone analogs of the hormone 1alpha,25-dihydroxyvitamin D3. AB - A series 2a-4b of seven new side-chain ketone analogs of calcitriol (1) have been prepared. Unexpectedly, several of these 24- and 25-tert-butyl ketones, even though lacking the classical side-chain tertiary hydroxyl group, are considerably more antiproliferative in vitro than the hormone calcitriol (1) even at physiologically relevant low nanomolar concentrations and are less calcemic than calcitriol (1) in vivo. In addition, ketone analog 19-nor-2a is not significantly less calcemic in vivo than 19-methylene analog 2a. PMID- 16039133 TI - Stereochemical determination and bioactivity assessment of (S)-(+)-curcuphenol dimers isolated from the marine sponge Didiscus aceratus and synthesized through laccase biocatalysis. AB - Electrospray ionization mass spectrometry-guided isolation of extracts from Didiscus aceratus led to the discovery of several new derivatives of the bioactive bisabolene-type sponge metabolite (S)-(+)-curcuphenol (1). The compounds obtained by this method included a mixture of known (2) and new (3) dihydroxylated analogs as well as a novel family of dimeric derivatives, dicurcuphenols A-E (4-8), and dicurcuphenol ether F (9). Dimers 4-9 were also subsequently obtained through a hemisynthetic method in which 1 was incubated with the enzyme laccase. Atropisomeric dimers 5 and 6 were subjected to vibrational circular dichroism analysis thereby establishing their absolute biaryl axial chirality as P and M, respectively. In contrast to 1, metabolites 2 9 exhibited weak or no cytotoxic or lipoxygenase inhibitory effects. PMID- 16039134 TI - A comparison of the high count-rate performance of three commercially available digital signal processors. AB - Three commercial gamma-ray digital signal processors, a Canberra InSpector 2000, an ORTEC DigiDART, and an X-ray Instrumentation Associates Polaris system, coupled to a Canberra 2002C resistive-feedback preamplifier-equipped high-purity germanium detector, were performance tested to input rates of 440 kHz. The spectrometers were evaluated on their throughput, stability and peak shape performance. The accuracy of their quantitative corrections for dead time and pile-up were also tested. All three of the tested units performed well at input rates that strain most analog spectroscopy systems. PMID- 16039135 TI - Application of gamma-ray computed tomography to analysis of soil structure before density evaluations. AB - Several studies have shown that physical properties of a soil may vary significantly over relatively short distances. This variability is due to the nature of the soil, agricultural management practices and sampling procedures; this study is focused on the latter factor. gamma-Ray tomography was used as a tool to evaluate the quality of soil samples collected for estimating physical characteristics of the soil and to detect possible damage to the soil in the process of sampling. A first-generation tomograph with an 241Am source and a 3''x3'' NaI(Tl) scintillation crystal detector coupled to a photomultiplier tube was employed. The results show that computed tomography can provide an insight into the sample structure, which helps to select samples that are best suited for evaluation of physical characteristics of a soil. PMID- 16039136 TI - Colorectal cancer adjuvant treatment in elderly patients. AB - Colorectal adenocarcinoma ranks second as a cause of death due to cancer in the Western world. In Europe, 40% of patients with this disease is over 70 years old and only 52% of them with positive nodes usually receive an adjuvant chemotherapy. Despite early reports to the contrary, these patients tolerate cancer treatment and surgical resections as well as their younger counterparts but as a result of exclusion criteria, those receiving an adjuvant therapy are very few. This paper examines the factors pertinent to the small number of clinical trials designed for adjuvant colorectal cancer in the elderly. PMID- 16039137 TI - Expression patterns of PDGF-A, -B, -C and -D and the PDGF-receptors alpha and beta in activated rat hepatic stellate cells (HSC). AB - The platelet-derived growth factor (PDGF) family, which regulates many physiological and pathophysiological processes has recently been enlarged by two new members, the isoforms PDGF-C and -D. Little is known about the expression levels of these new members in hepatic fibrosis. We therefore investigated by quantitative real time PCR (Taqman) the mRNA expression profiles of all four PDGF isoforms in transdifferentiating primary cultured hepatic stellate cells (HSC), an in vitro model system of hepatic fibrogenesis, either with or without stimulation of the cells with PDGF-BB or TGF-beta1. All four isoforms were expressed in HSC transdifferentiating to myofibroblast-like cells (MFB) albeit with different profiles: while PDGF-A mRNA exhibited minor fluctuations only, PDGF-B was rapidly down-regulated. In contrast, both PDGF-C and -D mRNA were strongly induced: PDGF-C up to 5 fold from day 2 to day 8 and PDGF-D up to 8 fold from day 2 to day 5 of culture. Presence of PDGF-DD in activated HSC was confirmed at the protein level by immunocytochemistry. Stimulation of HSC and MFB with PDGF-BB led to down-regulation of the new isoforms, whereas TGF-beta1 upregulated PDGF-A only. We further show that PDGF receptor-beta (PDGFR-beta) mRNA was rapidly upregulated within the first day of culture and was constantly expressed from day 2 on while the expression profile of PDGFR-alpha mRNA was very similar to that of PDGF-A during transdifferentiation. Given the dramatic changes in PDGF-C and -D expression, which may compensate for down-regulation of PDGF-B, we hypothesize that the new PDGF isoforms may fulfil specific functions in hepatic fibrogenesis. PMID- 16039138 TI - Increase in plasma homocysteine levels induced by drug treatments in neurologic patients. AB - Homocysteine (Hcy) is a thyol amino acid resulting from de-methylation of methionine, an essential amino acid derived from dietary proteins. It is metabolized through two pathways: re-methylation and transsulfuration, which use as cofactors folate, vitamin B6 and vitamin B12. Hyperhomocysteinemia has been identified as a risk factor for cerebrovascular disease, dementia, impaired cognitive function and depression. Several drugs may interfere with metabolic pathways of Hcy, leading to an alteration of plasma Hcy levels. Lipid-lowering agents, used to reduce the risk of cerebral venous thrombosis or occlusive vascular disease in patients with high levels of plasmatic lipids, can increase plasma Hcy levels. Hyperhomocysteinemia has been also documented in Parkinson disease patients treated with levodopa and in epileptic patients after chronic treatment with antiepileptic drugs. In contrast, vitamins supplementations may be warranted in patients treated with lipid-lowering agents, levodopa and antiepileptic drugs in order to maintain normal plasma Hcy values. In contrast, higher doses of vitamins can induce dysfunctions in central and peripheral nervous system; therefore excessive supplements should be avoided. PMID- 16039139 TI - Over-expression and purification of isotopically labeled recombinant ligand binding domain of orphan nuclear receptor human B1-binding factor/human liver receptor homologue 1 for NMR studies. AB - The human hepatitis B virus enhancer II B1 binding factor (hB1F), which regulates the expression of hepatitis B virus genes, is identified as a nuclear receptor. It regulates several liver-specific genes and plays an important role in the bile acid biosynthesis pathway. A significantly optimized protocol has been worked out to prepare 15N and/or 13C-labeled hB1F ligand-binding domain in minimal medium with high yields for NMR studies. Under the various conditions optimized for the purification of His6-hB1F ligand-binding domain, the yield of the purified protein is estimated to be 25-30 mg from 0.5 L of M9 minimal media. Electrospray ionization mass spectrometry data confirm the correctness of the primary sequence. Dynamic light scattering experiment proves that the protein exists as a monomeric form. In addition, the circular dichroism results show that the protein has a well-regulated secondary structure and a high alpha-helical content in ammonium bicarbonate buffer at 20 degrees C and pH 7.4. Finally, uniformly 15N labeled protein is characterized by a TROSY-HSQC spectrum, and the dispersion of 15N-1H cross-peaks in the spectrum indicates the presence of well-ordered and properly folded protein as a monomer. PMID- 16039140 TI - Baculovirus expression, purification, and characterization of human protein phosphatase 2A catalytic subunits alpha and beta. AB - Protein phosphatase 2A (PP2A) contains a 36-kDa catalytic subunit (PP2Ac), a 65 kDa structural subunit (PR65/A), and a regulatory B subunit. The core enzyme consists of the structural and catalytic subunits. The catalytic subunit exists as two closely related isoforms, alpha and beta. Several natural toxins, including okadaic acid (OA) and microcystins, specifically inhibit PP2A. To obtain biologically active recombinant PP2A and to compare the properties of the PP2A catalytic subunit alpha and beta isoforms, we expressed human PP2Acalpha and cbeta in High Five insect cells. The recombinant PP2Acalpha and cbeta possess similar phosphatase activities using p-NPP and phosphopeptide as substrates and are strongly inhibited by OA and microcystin-LR to similar degrees. In addition, PP2Acalpha or cbeta was co-expressed with PR65/A and co-purified as a core dimer, PP2AD (Aalpha/calpha and Aalpha/cbeta) with PR65alpha/Aalpha. The recombinant PP2AD bound to the B subunit in vitro. These results show that the recombinant PP2Acalpha and cbeta are identical in their ability to associate with the A and B subunits, in their phosphatase activities, and in carboxyl-methylation. Furthermore, our results show that High Five insect cells can produce biologically active recombinant PP2A, which should be a valuable tool for detecting natural toxins and investigating the mechanism of PP2A catalysis and other protein interactions. PMID- 16039141 TI - Overexpression and characterization of a lipase from Bacillus subtilis. AB - A novel plasmid, pBSR2, was constructed by incorporating a strong lipase promoter and a terminator into the original pBD64. A mature lipase gene from Bacillus subtilis strain IFFI10210, an existing strain for lipase expression, was cloned into the plasmid pBSR2 and transformed into B. subtilis A.S.1.1655. Thus, an overexpression strain, BSL2, was obtained. The yield of lipase is about 8.6 mg protein/g of wet weight of cell mass and 100-fold higher than that in B. subtilis strain IFFI10210. The recombinant lipase was purified in a three-step procedure involving ammonium sulfate fractionation, ion exchange, and gel filtration chromatography. Characterizations of the purified enzyme revealed a molecular mass of 24 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, maximum activity at 43 degrees C and pH 8.5 for hydrolysis of p-nitrophenyl caprylate. The values of Km and Vm were found to be 0.37 mM and 303 micromol mg-1 min-1, respectively. The substrate specificity study showed that p-nitrophenyl caprylate is a preference of the enzyme. The metal ions Ca2+, K+, and Mg2+ can activate the lipase, whereas Fe2+, Cu2+, and Co2+ inhibited it. The activity of the lipase can be increased about 48% by sodium taurocholate at the concentration of 7 mM and inhibited at concentrations over 10 mM. PMID- 16039142 TI - Effect of multiple injections of beta-glucan on non-specific immune response and disease resistance in Labeo rohita fingerlings. AB - The purpose of this study was to determine if multiple injections of different dosages of beta-glucan derived from barley would enhance the immune response and disease resistance against infections due to opportunistic pathogens Aeromonas hydrophila and Edwardsiella tarda in Labeo rohita fingerlings. Hence, four different dosages of beta-glucan suspension in phosphate-buffered saline at the rate of 0, 5, 10, 15 mg kg(-1) body weight of fish were injected intraperitoneally to the fingerlings of Labeo rohita at two-week intervals for four times. After every two-week interval different serum biochemical, haematological and immunological parameters of fish were evaluated. At the end of immunostimulation trial of 56 days, fish were divided into four subgroups under each major treatment group for challenge through i.p injection and bath immersion with two pathogens, A. hydrophila and E. tarda. The mortality (%) and agglutinating antibody titre was recoded on 28th day post challenge. Most of the immune parameters such as leucocyte count, phagocytic ratio, phagocytic index, lysozyme activity, complement activity, serum bactericidal activity were significantly (P < 0.05) enhanced on 42 days after three i.p injections of 10 mg of beta-glucan kg(-1) body wt. Challenge study indicated least mortality in the group of fishes injected with medium dose of 10 mg of beta-glucan kg(-1) body wt. four times. Multiple injections of beta-glucan might have maintained the activation of phagocytic cells for a long period which in turn would lead to long term protection in fishes. Thus, injections of 10 mg of beta-glucan kg(-1) body wt. for three times can be advocated to enhance the immune response of fish species under aquaculture. PMID- 16039143 TI - Attitudes to the right- and left: frontal ERP asymmetries associated with stimulus valence and processing goals. AB - We used dense-array event-related potentials (ERP) to examine the time course and neural bases of evaluative processing. Participants made good vs. bad (evaluative) and abstract vs. concrete (nonevaluative) judgments of socially relevant concepts (e.g., "murder," "welfare"), and then rated all concepts for goodness and badness. Results revealed a late positive potential (LPP) beginning at about 475 ms post-stimulus and maximal over anterior sites. The LPP was lateralized (higher amplitude and shorter latency) on the right for concepts later rated bad, and on the left for concepts later rated good. Moreover, the degree of lateralization for the amplitude but not the latency was larger when participants were making evaluative judgments than when they were making nonevaluative judgments. These data are consistent with a model in which discrete regions of prefrontal cortex (PFC) are specialized for the evaluative processing of positive and negative stimuli. PMID- 16039144 TI - Localization of human supratemporal auditory areas from intracerebral auditory evoked potentials using distributed source models. AB - While source localization methods are increasingly developed to identify brain areas underlying scalp electro/magnetoencephalographic data (EEG/MEG), these methods have not yet been used to identify the sources of intracerebral signals which offer highly detailed information. Here, we adapted the minimum current estimates method to intracranial data in order to localize supratemporal sources of intracerebral auditory 1-kHz-tone-evoked potentials occurring within 100 ms after stimulus onset. After an evaluation of localization method and despite inter-subject variability, we found a common spatiotemporal pattern of activities, which involved the first Heschl's gyrus (H1) and sulcus (HS), the Planum Temporale (PT), H2/H3 when present, and the superior temporal gyrus (STG). Four time periods of activity were distinguished, corresponding to the time range of the scalp components P0, Na, Pa/Pb, and N100. The sources of the earliest components P0 (16-19 ms) and Na (20-25 ms) could be identified in the postero medial portion of HS or H1. Then, several areas became simultaneously active after 25 ms. The Pa/Pb time range (30-50 ms) was characterized by a medio-lateral and postero-anterior propagation of activity over the supratemporal plane involving successively H1/HS, the Planum Temporale, H2/H3 when present, and the STG. Finally, we found to a large extent that the N100 (55-100 ms) involved almost the same areas as those active during the Pa/Pb complex, with a similar propagation of activities. Reconstructing scalp data from these sources on fictive EEG/MEG channels reproduced classical auditory evoked waveforms and topographies. In conclusion, the spatiotemporal pattern of activation of supratemporal auditory areas could be identified on the individual anatomy using current estimates from intracerebral data. Such detailed localization approach could also be used prior to epilepsy surgery to help identify epileptogenic foci and preserve functional cortical areas. PMID- 16039145 TI - Molecular and morphological phylogenetic analysis of Brachiaria and Urochloa (Poaceae). AB - The taxonomic relationships of Brachiaria and Urochloa have been questioned based on previous morphological studies. In this paper, we reconsider the phylogenetic relationships of these genera using 22 species of Brachiaria and Urochloa and six species of Paniceae as out-groups. The ITS1, 5.8S, and ITS2 region (internal transcribed spacer) of nuclear ribosomal DNA and eight morphological characters of the inflorescence were compiled into a data matrix. The cladistic analyses suggest that Urochloa-Brachiaria as a complex is paraphyletic with Eriochloa and Melinis. Species of all these genera share molecular synapomorphies and belong to the same monophyletic groups. The results confirm the continuous gradation between those genera previously found in several morphological studies. Therefore, the following eight new combinations are made: Urochloa bovonei (Chiov.) A.M. Torres & C.M. Morton, Urochloa dura (Stapf) A.M. Torres & C.M. Morton, Urochloa dura var. dura (Stapf) A.M. Torres & C.M. Morton, Urochloa dura var. pilosa (J.G. Anderson) A.M. Torres & C.M. Morton, Urochloa lachnantha (Hochst.) A.M. Torres & C.M. Morton, Urochloa leersioides (Hochst.) A.M. Torres & C.M. Morton, Urochloa nigropedata (Munro ex Ficalho & Hiern) A.M. Torres & C.M. Morton, and Urochloa subulifolia (Mez) A.M. Torres & C.M. Morton. PMID- 16039146 TI - Phylogenetic relationships and biogeography of Podarcis species from the Balkan Peninsula, by bayesian and maximum likelihood analyses of mitochondrial DNA sequences. AB - Wall lizards of the genus Podarcis (Sauria, Lacertidae) comprise 17 currently recognized species in southern Europe, where they are the predominant nonavian reptile group. The taxonomy of Podarcis is complex and unstable. Based on DNA sequence data, the species of Podarcis falls into four main groups that have substantial geographic coherence (Western island group, southwestern group, Italian group, and Balkan Peninsula group). The Balkan Peninsula species are divided into two subgroups: the subgroup of P. taurica (P. taurica, P. milensis, P. gaigeae, and perhaps P. melisellensis), and the subgroup of P. erhardii (P. erhardii and P. peloponnesiaca). In the present study, the question of phylogenetic relationships among the species of Podarcis encountered in the Balkan Peninsula was addressed using partial mtDNA sequences for cytochrome b (cyt b) and 16S rRNA (16S). The data support the monophyly of Podarcis and suggest that there are three phylogenetic clades: the clade A (P. taurica, P. gaigeae, P. milensis, and P. melisellensis); the clade B (P. erhardii and P. peloponnesiaca), and the clade C (P. muralis and P. sicula). By examining intraspecific relationships it was found that extant populations of P. erhardii are paraphyletic. Furthermore, subspecies previously defined on the basis of morphological characteristics do not correspond to different molecular phylogenetic clades, suggesting that their status should be reconsidered. The distinct geographic distribution of the major clades of the phylogenetic tree and its topology suggest a spatial and temporal sequence of phylogenetic separations that coincide with some major paleogeographic separations during the geological history of the Aegean Sea. The results stress the need for a reconsideration of the evolutionary history of Balkan Podarcis species and help overcome difficulties that classical taxonomy has encountered at both the species and subspecies level. PMID- 16039147 TI - Mitochondrial phylogeny of pine cone beetles (Scolytinae, Conophthorus) and their affiliation with geographic area and host. AB - Pine cone beetles (Conophthorus spp.) feed and kill immature cones of Pinus species, thereby reducing seed production and seriously impairing reforestation of forest ecosystems. Population variation of Conophthorus reproductive behavior has hampered the development of semiochemical control of these pests. This difficulty is compounded by a lack of taxonomic knowledge and species diagnostic characters. Researchers and managers rely, in part, on host associations and geographic locality for species identifications and these have arguable taxonomic utility. However, host use and/or geographic separation may influence Conophthorus lineage diversification. To improve Conophthorus taxonomy and understand the association of host and geography with lineage diversification, a phylogeny of 43 individuals, including all valid species and a robust sample of C. ponderosae from different hosts, is reconstructed using 785 nucleotides of the 3'-end of the mitochondrial cytochrome oxidase I gene. Thirty trees were recovered in a parsimony analysis and the strict consensus was well resolved and supported by branch support measures. Conophthorus was monophyletic but mitochondrial polyphyly was uncovered for several species. The data also suggested an underestimation of species diversity. Phylogenetically related Conophthorus lineages were significantly associated with geographic proximity but not with host, as indicated by comparisons of character optimized geographic distributions and host associations against randomized distributions of these attributes on the parsimony tree. These results suggest that geographic separation better explains the mode of Conophthorus lineage diversification than does host specialization. Based on these results, researchers and managers of Conophthorus should consider populations as potentially different evolutionary entities until species boundaries are delineated via a robust phylogenetic revision of Conophthorus. PMID- 16039148 TI - Multiple host shifts between distantly related plants, Juglandaceae and Ericaceae, in the leaf-mining moth Acrocercops leucophaea complex (Lepidoptera: Gracillariidae). AB - Insect herbivores such as gall formers and leaf miners are often highly specialized and adapted to their respective natal host plants. Due to the specialization and adaptation, it is presumed that host shifts readily occur among closely related plant species. Leaf-mining moths, the Acrocercops leucophaea complex, consist of three species, A. leucophaea, A. defigurata, and A. transecta. Larvae of all the species of the complex feed on Juglandaceae plants, but A. leucophaea and A. transecta are also associated with an Ericaceae plant, which is quite distantly related to Juglandaceae. Such a host utilization as in this species complex is very rare among phytophagous insects. In the present study, we estimate the history of host shifts by reconstructing the phylogeny of the A. leucophaea complex using molecular data (partial sequence of mitochondrial COI, 12S rDNA, and ND5). Parsimony and maximum likelihood analyses indicated that the common ancestor of the A. leucophaea complex used Juglandaceae only, and that the association with Ericaceae has evolved in A. leucophaea and A. transecta independently. Parametric bootstrap analysis also supported multiple origins of the association with Ericaceae in this complex. These results imply that there are ecological and biochemical factors that promote host shifting between Juglandaceae and Ericaceae despite the two families being not closely related. PMID- 16039149 TI - A phylogeny of the schoenoid sedges (Cyperaceae: Schoeneae) based on plastid DNA sequences, with special reference to the genera found in Africa. AB - Despite its large size (about 700 species), the australy-centred sedge tribe Schoeneae has received little explicit phylogenetic study, especially using molecular data. As a result, generic relationships are poorly understood, and even the monophyly of the tribe is open to question. In this study, plastid DNA sequences (rbcL, trnL-trnF, and rps16) drawn from a broad array of Schoeneae are analysed using Bayesian and parsimony-based approaches to infer a framework phylogeny for the tribe. Both analytical methods broadly support the monophyly of Schoeneae, Bayesian methods doing so with good support. Within the schoenoid clade, there is strong support for a series of monophyletic generic groupings whose interrelationships are unclear. These lineages form a large polytomy at the base of Schoeneae that may be indicative of past radiation, probably following the fragmentation of Gondwana. Most of these lineages contain both African and non-African members, suggesting a history of intercontinental dispersal. The results of this study clearly identify the relationships of the African-endemic schoenoid genera and demonstrate that the African-Australasian genus Tetraria, like Costularia, is polyphyletic. This pattern is morphologically consistent and suggests that these genera require realignment. PMID- 16039150 TI - Evolutionary history of freshwater sculpins, genus Cottus (Teleostei; Cottidae) and related taxa, as inferred from mitochondrial DNA phylogeny. AB - The freshwater sculpins, genus Cottus (Teleostei; Cottidae), comprise bottom dwelling fishes that exhibit various life-history styles, having radiated throughout Northern Hemisphere freshwater habitats. The phylogenetic relationships among Cottus and related taxa were estimated from mitochondrial DNA 12S rRNA and control region (CR) sequences, the freshwater sculpins examined falling into five lineages (A-E). Lineage A consisted of Trachidermus fasciatus and C. kazika, both having a catadromous life-history. The remaining species (lineages B-E) spawn in freshwater habitats regardless of life-history (amphidromous, lacustrine or fluvial), suggesting that the various life-history types post-dated a common ancestor of lineages B-E. Molecular clock estimates suggested a Pliocene-Pleistocene radiation (or Miocene-Pliocene from the alternative clock) of lineages B-E. In eastern Eurasia, speciation with life history changes to amphidromous or fluvial styles has apparently occurred independently in some lineages, as a general pattern. Mitochondrial DNA CR phylogeny showed the monophyletic Baikalian cottoids (Cottoidei) to be nested within Cottus and Trachidermus, suggesting that the former ecologically and morphologically divergent cottoids may have originated from a single lineage which invaded the ancient lake. PMID- 16039151 TI - Inducible nitric oxide synthase involvement in the mechanism of action of Saccharomyces boulardii in castor oil-induced diarrhoea in rats. AB - The biotherapeutic agent Saccharomyces boulardii has been shown to inhibit castor oil-induced diarrhoea in rats in a dose-response fashion, and one of the suggested mechanisms of action included involvement of the nitric oxide pathway. The present study was designed to address this mechanism of action by firstly measuring the effects of S. boulardii on the inducible nitric oxide synthase (iNOS) isoform activity in vitro. Second, the effects of S. boulardii on the increase in colonic citrulline level associated with castor oil treatment were examined. In vitro, S. boulardii showed a dose-dependent inhibition of iNOS activity with an IC50 of 0.89 mg/ml. In the rat diarrhoea model, the antidiarrhoeal effect of S. boulardii was confirmed using a single oral dose of 12 x 10(10) CFU/kg (viable cells). In this model, castor oil significantly elevated citrulline level from 2526+/-164 to 3501+/-193 nmol/g in the colon. When the rats were treated with the same antidiarrhoeal single dose of S. boulardii, no increase in citrulline level was observed. Moreover, the iNOS inhibitor 1400 W at 10 mg/kg and the inhibitor of iNOS expression dexamethasone at 1 mg/kg, administered subcutaneously, blocked the citrulline production induced by the laxative. Taken together, these findings confirm the involvement of inhibition of the inducible isoform of nitric oxide synthase in the mechanism of action of S. boulardii in diarrhoea. PMID- 16039152 TI - Contribution of biofilm regulatory protein A of Streptococcus mutans, to systemic virulence. AB - Streptococcus mutans is occasionally isolated from the blood of patients with bacteremia and infective endocarditis (IE), and the possibility that it could be pathogenic for those diseases has been discussed. The initial important step for the involvement of bacterial pathogens in the virulence of IE is thought to be survival in blood for an extended period. Recently, the brpA gene encoding biofilm regulatory protein A (BrpA) of S. mutans was cloned and sequenced, after which it was shown that inactivation of brpA in an isogenic mutant strain resulted in longer chain formation than in the parental strain. In the present study, a BrpA-defective isogenic mutant strain (MT8148BRD) was constructed from strain MT8148. In an analysis of its susceptibility to phagocytosis by human polymorphonuclear leukocytes (PMNs), the phagocytosis rate of MT8148BRD was shown to be significantly lower than that of MT8148 (P < 0.01). Next, strains with various chain lengths were produced by culturing MT8148 in media with various initial pH levels, which revealed that there was a statistically negative correlation between phagocytosis susceptibility and chain length (P < 0.01). Further, MT8148BRD was found to possess higher platelet aggregation properties than MT8148 (P < 0.05). In addition, injection of MT8148BRD into the jugular vein of specific pathogen-free Sprague-Dawley rats resulted in a longer duration of bacteremia, which prolonged systemic inflammation for a longer period than in those infected with MT8148. These results indicate that S. mutans BrpA is associated with virulence in blood, due to its correlation to phagocytosis susceptibility and platelet aggregation properties. PMID- 16039153 TI - Phloem and xylem specification: pieces of the puzzle emerge. AB - The plant vascular system is composed of two tissue types, xylem and phloem, which originate from the vascular meristem, the procambium. Recently, several regulatory mechanisms that control the specification of these two tissue types have been uncovered. These include the asymmetric patterning of xylem and phloem in the vascular bundle by the class III HD-ZIP and KANADI genes, the tissue-type specific control of vascular cell proliferation by brassinosteroids and class III HD-ZIP genes, the regulation of vascular tissue identity by the MYB-like transcription factor APL, and inductive signalling during xylem differentiation by xylogen. These findings define an emerging developmental framework for the control of vascular tissue specification. PMID- 16039154 TI - Regulated proteolysis in light signaling. AB - Photoreceptors regulate many aspects of development throughout the life cycle of a plant. Recent advances have demonstrated the importance of ubiquitin-mediated proteolysis in the control of development by light. Both some of the photoreceptors themselves and, in particular, transcription factors that are involved in transducing the light signal are subject to regulated ubiquitination and subsequent degradation by the 26S proteasome. PMID- 16039155 TI - Signalling and gene regulation in response to ultraviolet light. AB - In contrast to phytochrome-, cryptochrome- and phototropin-sensing systems, about which considerable knowledge has accumulated, the ultraviolet-B (UVB) photoreceptor is not yet known at the molecular level. Information about the downstream signalling events that underlie UVB-provoked physiological responses is limited. Recent whole-genome transcript profiling, isolation of mutants that are impaired in specific UVB-induced responses and detailed photobiological studies suggest that responses that are triggered by shorter wavelength UVB and longer wavelength UVB are mediated by two different sensory systems. The bZIP transcription factor HY5 was recently identified as an important player in the long-wavelength UVB-induced signal transduction cascade. Advances in the development of luciferase-reporter lines will make it feasible to perform high throughput genetic screens to isolate novel mutants that are impaired in sensing or transducing signals downstream of the putative UVB photoreceptor(s). PMID- 16039156 TI - Impact of low doses of tritium on the marine mussel, Mytilus edulis: genotoxic effects and tissue-specific bioconcentration. AB - Despite growing scientific, public and regulatory concern over the discharge of radioactive substances, no serious attempts have been made to develop a rationale to evaluate the impact of environmentally relevant radionuclides in the aquatic environment. In this study, we have evaluated the genotoxic effects and tissue specific concentration of tritium (added as tritiated water, HTO) in the adult life stage of the edible mussel, Mytilus edulis. The genotoxic effects were quantified in terms of the induction of: (a) micronuclei (MN), and (b) DNA single strand breaks/alkali-labile sites using alkaline single-cell gel electrophoresis (Comet assay) in the haemocytes of exposed animals. The assays were optimised and validated using a range of concentrations (18-56 mgl(-1)) of ethylmethane sulfonate (EMS), a direct-acting reference genotoxic agent, over different exposure periods. Mussels were exposed to a series of concentrations of HTO equivalent to a dose range from 12 to 485 muGyh(-1) for 96 h, and different tissues and organs were then extracted and analysed. The study revealed a dose dependent increase in the response for both the MN test and the Comet assay and for both EMS and HTO. In addition, HTO delivering dose rates below 500 muGyh(-1) was shown to be capable of inducing genetic damage in the haemocytes of these bivalves. The study also showed that inorganic tritium accumulated differentially in mussel tissues in a dose-dependent manner, with the gut accumulating the highest amount of radioactivity, followed by the gill, mantle, muscle, foot and byssus thread. The faeces and pseudo-faeces accumulated least radioactivity over the exposure period. Differential accumulation of radionuclides has significant implications for biomonitoring programmes, for this and other aquatic organisms. The study also suggests that the generic dose limits recommended by the International Atomic Energy Agency for the protection of aquatic biota might not be applicable to all aquatic organisms. PMID- 16039157 TI - Linking signalling pathways, thymic stroma integrity and autoimmunity. AB - Medullary thymic epithelial cells (mTECs) are indispensable for self-tolerance to peripheral organs by virtue of their expression of a host of tissue-restricted self-antigens. The full extent of this promiscuous gene expression is confined to functionally mature mTECs. Consequently, any interference with signalling pathways directing the differentiation and/or proliferation of this mature subset will affect the scope of central tolerance and potentially predispose to autoimmunity. In a recent study, tumour necrosis factor receptor-associated factor 6 (TRAF6) has been identified as an essential component of a new signalling pathway directing mTEC development. PMID- 16039158 TI - Measuring Ag-specific immune responses: understanding immunopathogenesis and improving diagnostics in infectious disease, autoimmunity and cancer. AB - Characterization of antigen-specific immune responses at the single-cell level has been made possible by recent advancements in reagent and technology development, combined with increasing knowledge of molecular mechanisms. Fluorescently labelled MHC-peptide multimers and antigens identify directly specific T and B cells, respectively, whereas dynamic assays exploit mediator production or secretion, or the changes in surface expression of other proteins, to identify specific lymphocytes--some techniques enabling the recovery of viable cells. Meanwhile, multiparameter flow cytometry has emerged as the most versatile platform for integrating most of these methods. As the complexity of experimental data increases, so does the level of technical sophistication required for analysis and interpretation, both in terms of basic research and modern medicine, with new applications for infectious diseases, autoimmunity and cancer. PMID- 16039159 TI - Nitric oxide bioavailability in malaria. AB - Rational development of adjunct or anti-disease therapy for severe Plasmodium falciparum malaria requires cellular and molecular definition of malarial pathogenesis. Nitric oxide (NO) is a potential target for such therapy but its role during malaria is controversial. It has been proposed that NO is produced at high levels to kill Plasmodium parasites, although the unfortunate consequence of elevated NO levels might be impaired neuronal signaling, oxidant damage and red blood cell damage that leads to anemia. In this case, inhibitors of NO production or NO scavengers might be an effective adjunct therapy. However, increasing amounts of evidence support the alternate hypothesis that NO production is limited during malaria. Furthermore, the well-documented NO scavenging by cell free plasma hemoglobin and superoxide, the levels of which are elevated during malaria, has not been considered. Low NO bioavailability in the vasculature during malaria might contribute to pathologic activation of the immune system, the endothelium and the coagulation system: factors required for malarial pathogenesis. Therefore, restoring NO bioavailability might represent an effective anti-disease therapy. PMID- 16039160 TI - A glycine-cleavage complex as part of the folate one-carbon metabolism of Plasmodium falciparum. AB - The glycine-cleavage complex (GCV) and serine hydroxymethyltransferase represent the two systems of one-carbon transfer that are employed in the biosynthesis of active folate cofactors in eukaryotes. Although the understanding of this area of metabolism in Plasmodium falciparum is still at an early stage, we discuss evidence that genes and transcription products of the GCV are present and expressed in this parasite. The potential role of the GCV and its relevance to the life cycle and pathogenesis of the malaria erythrocytic stages are also considered. According to its expression profile, the GCV seems to be particularly active in gametocytes. The GCV enzyme dihydrolipoamide dehydrogenase has two isoforms encoded by two different genes. It has been demonstrated recently that both genes are functional, with one of them identified as being part of a pyruvate dehydrogenase complex that is present exclusively in the apicoplast of Plasmodium species. The other isoform probably forms part of the Plasmodium GCV. The GCV is the first enzyme complex involved in folate metabolism in this parasite that can be assumed, with a good degree of certainty, to be located in the mitochondria. PMID- 16039161 TI - A comprehensive and comparative analysis for MALDI FTMS lipid and phospholipid profiles from biological samples. AB - Described here is a computationally automated method for translating complex accurate mass spectra into biologically relevant and meaningful data. Rapid profiling of detailed high resolution mass spectra resulting from direct analysis of whole cells and tissues by matrix-assisted laser desorption/ionization (MALDI) Fourier transform mass spectrometry (FTMS) is discussed. Lipid and phospholipid ions create complex spectra containing multiple m/z values corresponding to the same fundamental chemical species. A computational approach is employed to sort ions, with mass to charge ratios lower than m/z 1000, into groups of similar lipid and phospholipid compositions for comprehensive and rapid analysis. By sorting or binning ions in this manner, variations in the degree of cation exchange can be avoided, thus increasing the comparability of the data. The result is displayed as a histogram that is easily interpretable and comparable with similar analyses and is particularly useful for direct comparison of similar tissues. Spectra of leaves from a healthy Prunus persica (peach) tree are compared with those from leaves infected by the fungus Taphrina deformans. Although the infection can be seen as a difference in leaf structure and by visual inspection of the mass spectra, the method described here details the chemical difference in phospholipid compositions and their relative abundances. PMID- 16039162 TI - Cyclophilins in rheumatoid arthritis--stepping into an undiscovered country? PMID- 16039163 TI - Alteration of splicing signals in a genomic/cDNA hybrid VEGF gene to modify the ratio of expressed VEGF isoforms enhances safety of angiogenic gene therapy. AB - Vascular endothelial growth factor (VEGF)-mediated physiological angiogenesis results from the concerted action of three major VEGF isoforms (VEGF121, 165, 189), which arise from alternate splicing. We have previously shown that expression of a mixture of VEGF isoforms via gene transfer is considerably more potent than expression of a single VEGF isoform. To test the hypothesis that different mixtures of VEGF isoforms may offer the same therapeutic benefit with a better safety profile, we compared the efficacy and safety of an adenovirus gene transfer vector expressing the three major VEGF isoforms (AdVEGF-All) in the normal ratio to those of AdVEGF-All6A+, in which the splicing sequences for exon 6A were altered to promote expression of VEGF189 at the expense of VEGF121. Both vectors were equally potent in mediating recovery of hind-limb blood flow following experimental ischemia. By contrast, intravenous administration of AdVEGF-All6A+ yielded enhanced survival and a lower capacity to support tumor growth compared to AdVEGF-All, and intratracheal administration of AdVEGF-All6A+ resulted in less pulmonary edema than that of AdVEGF-All. We conclude that AdVEGF All and AdVEGF-All6A+ are similar in potency but that AdVEGF-All6A+ is safer. This suggests that AdVEGF-All6A+ may be the preferred candidate for clinical development. PMID- 16039164 TI - Targeting ECM-integrin interaction with liposome-encapsulated small interfering RNAs inhibits the growth of human prostate cancer in a bone xenograft imaging model. AB - The intricate intracellular communication between stromal and epithelial cells, which involves cell-cell-, cell-insoluble extracellular matrix- (ECM), and cell soluble factor-mediated signaling processes, is an attractive target for therapeutic intervention in hormone-refractory and bone-metastatic prostate cancer. In the present study we demonstrated that androgen-independent PC3 prostate cancer cells adhered to and migrated on vitronectin (VN), a major noncollagenous ECM in mature bone, through the expression of alphav-containing integrin receptors alphavbeta1 and alphavbeta5 on the cell surface, as determined by antibody function blocking assay and flow cytometry analysis. Small interfering RNAs (siRNAs) targeting human integrin alphav markedly reduced their respective mRNA and protein expression in cells, resulting in nearly complete reduction in VN-mediated cancer progression in vitro. In vivo quantitative bioluminescence analysis of human prostate cancer bone xenografts demonstrated for the first time that intratumoral administration of liposome-encapsulated human alphav-siRNAs significantly inhibits the growth of luciferase-tagged PC3 tumors in skeleton, which was associated with decreased integrin alphav expression and increased apoptosis in tumor cells. This integrin-based gene therapy is particularly suitable for the treatment of prostate cancer bone metastasis. PMID- 16039165 TI - Vitamin E prevents cell death induced by mild oxidative stress in chicken skeletal muscle cells. AB - Apoptosis and necrosis are two forms of cell death that can occur in response to various agents and oxidative damage. In addition to necrosis, apoptosis contributes to muscle fiber loss in various muscular dystrophies as well participates in the exudative diathesis in chicken, pathology caused by dietary deficiency of vitamin E and selenium, which affects muscle tissue. We have used chicken skeletal muscle cells and bovine fibroblasts to study molecular events involved in the cell death induced by oxidative stress and apoptotic agents. The effect of vitamin E on cell death induced by oxidants was also investigated. Treatment of cells with anti-Fas antibody (50 to 400 ng/mL), staurosporine (0.1 to 100 microM) and TNF-alpha (10 and 50 ng/mL) resulted in a little loss of Trypan blue exclusion ability. Those stimuli conducted cells to apoptosis detected by an enhancement in caspase activity upon fluorogenic substrates but this activity was not fully blocked by the caspase inhibitor Z-VAD-fmk. Oxidative stress induced by menadione (10, 100 and 250 muM) promoted a significant reduction in cell viability (10%, 20% and 35% for fibroblasts; 20%, 30% and 75% for muscle cells, respectively) and caused an increase in caspase activity and DNA fragmentation. H2O2 also promoted apoptosis verified by caspase activation and DNA fragmentation, but in higher doses induced necrosis. Vitamin E protected cells from death induced by low doses of oxidants. Although it was ineffective in reducing caspase activity in fibroblasts, this vitamin diminished the enzyme activity in muscle cells. These data suggested that oxidative stress could activate apoptotic mechanisms; however the mode of cell death will depend on the intensity and duration of the stimulus, and on the antioxidant status of the cells. PMID- 16039166 TI - Apoptosis and biochemical biomarkers of stress in spiders from industrially polluted areas exposed to high temperature and dimethoate. AB - The aim of this study was to evaluate the relations between apoptosis and the activity of antioxidant enzymes (superoxide dismutase; catalase) and quantitative changes in stress protein positive cells (Hsp70; metallothionein) in midgut glands of funnel web spiders Agelena labyrinthica (Agelenidae) and wolf spiders Pardosa lugubris (Lycosidae) exposed to high temperature and pesticide under laboratory conditions. The spiders were collected from two meadow ecosystems differently polluted with metals (Olkusz and Pilica, southern Poland). Under stress conditions, P. lugubris had fewer apoptotic cells in the midgut glands than A. labyrinthica. In P. lugubris from both sites, the observed increase in the percentage of metallothionein and Hsp70-positive cells, simultaneous with intensification of superoxide dismutase and catalase activity, suggests an anti apoptotic function of those proteins in representatives of wandering spiders. In the midgut glands of A. labyrinthica, heat shock and dimethoate increased the number of Annexin V-positive cells as well as the amounts of mitochondria with low transmembrane potential (DeltaPsi(m)) versus the control. The changes in the percentage of MT- and Hsp70-positive cells in funnel web spiders were less than in wolf spiders. The absence of change in SOD and CAT activity in A. labyrinthica shows that the participation of those enzymes in antioxidant reactions is minimal in this species. PMID- 16039167 TI - Application of polymaleimidostyrene as a convenient immobilization reagent of enzyme in biosensor. AB - Sulfhydryl groups of glucose oxidase (GOD) were reacted with maleimide groups of polymaleimidostyrene (PMS) which was coated onto the porous carbon sheet, and the carbon sheet immobilized by GOD was combined with an oxygen electrode to fabricate a glucose sensor. The activity of thiolated GOD immobilized to PMS is much larger than that of native GOD immobilized to PMS. The good linear relationship of glucose and oxygen current response was obtained in a concentration range from 0.1 to 2 mM and upper limit of linear range was found to be 3.0 mM. The immobilized GOD activity is highly dependent on pH at immobilization and the maximum activity was obtained at pH 5.5, probably because the SH groups of GOD that are indispensable for generation of enzyme activity is not exposed at this pH. It was found that PMS is very effective reagent to immobilize enzyme strongly via covalent bond, because high density of maleimide groups of PMS can catch not only exposed SH groups but also buried SH groups. PMID- 16039168 TI - On the sub-maximal yield and photo-electric stimulation of chlorophyll a fluorescence in single turnover excitations in plant cells. AB - A set of expressions is derived which quantifies the chlorophyll fluorescence yield in terms of rate constants of primary light reactions of PSII, the fraction of open and semi-open RCs and of the electric field sensed by the RC in the thylakoid membrane. The decay kinetics of the chlorophyll fluorescence yield after a single turnover excitation in the presence of DCMU show at least two components, one reversible within approx. 1 s and one with a dark reversion lasting more than 30 s. The latter is attributed to photochemical quenching; the fast component is interpreted to be associated at least partially with photo electrochemical control. It will be illustrated that (i) the sub-maximal fluorescence yield in single turnover excitation is associated with semi-closure of RCs, (ii) the trapping efficiency of semi-closed centers is less than 50% of that of open centers and (iii) the fluorescence yield of antennas with semi closed RCs has the highest sensitivity to changes in strength of photo-electric fields. PMID- 16039169 TI - Measurement of resiniferatoxin in serum samples by high-performance liquid chromatography. AB - A novel and simple method of extraction, separation, identification and quantification of resiniferatoxin (RTX) in serum samples is reported. Human serum and whole blood were treated with acetonitrile to denature proteins, such as orosomucoid, and the soluble fraction was passed through a reversed-phase C18 cartridge. RTX eluted from the cartridge was quantified by high-performance liquid chromatography (HPLC) using a reversed-phase C18 column. Reproducible recovery of RTX and tinyatoxin, an internal standard, from serum was achieved. Isocratic elution with 62% acetonitrile provided a suitable retention time without interfering peaks eluting near the analyte. Therefore, the procedure described provides a useful assay for determination of serum RTX pharmacokinetic parameters. PMID- 16039170 TI - Leptin regulates chondrocyte differentiation and matrix maturation during endochondral ossification. AB - Leptin has been suggested to mediate a variety of actions, including bone development, via its ubiquitously expressed receptor (Ob-Rb). In this study, we investigated the role of leptin in endochondral ossification at the growth plate. The growth plates of wild-type and ob/ob mice were analyzed. Effects of leptin on chondrocyte gene expression, cell cycle, apoptosis and matrix mineralization were assessed using primary chondrocyte culture and the ATDC5 cell differentiation culture system. Immunohistochemistry and in situ hybridization showed that leptin was localized in prehypertrophic chondrocytes in normal mice and that Ob-Rb was localized in hypertrophic chondrocytes in normal and ob/ob mice. Growth plates of ob/ob mice were more fragile than those of wild-type mice in a mechanical test and were broken easily at the chondro-osseous junction. The growth plates of ob/ob mice showed disturbed columnar structure, decreased type X collagen expression, less organized collagen fibril arrangement, increased apoptosis and premature mineralization. Leptin administration in ob/ob mice led to an increase in femoral and humeral lengths and decrease in the proportional length of the calcified hypertrophic zone to the whole hypertrophic zone. In primary chondrocyte culture, the matrix mineralization in ob/ob chondrocytes was stronger than that of wild-type mice; this mineralization in both types of mice was abolished by the addition of exogenous leptin (10 ng/ml). During ATDC5 cell differentiation culture, exogenous leptin at a concentration of 1-10 ng/ml (equivalent to the normal serum concentration of leptin) altered type X collagen mRNA expression and suppressed apoptosis, cell growth and matrix calcification. In conclusion, we demonstrated that leptin modulates several events associated with terminal differentiation of chondrocytes. Our finding that the growth plates of ob/ob mice were fragile implies a disturbance in the differentiation/maturation process of growth plates due to depletion of leptin signaling in ob/ob mice. These findings suggest that peripheral leptin signaling plays an essential role in endochondral ossification at the growth plate. PMID- 16039171 TI - Successful treatment of tachycardia-induced cardiomyopathy with LVAD in a 12-year old boy. AB - Tachycardia-induced cardiomyopathy is an unusual cardiac disease that is life threatening if tachycardia is not controlled. We report a 12-year-old boy who suffered from ectopic left atrial tachyarrhythmia that was refractory to medications and caused tachycardia-induced cardiomyopathy with severe heart failure. The patient required a left ventricular assist device (ABIOMED BVS5000 [ABIOMED Inc, Danvers, MA]) as a bridge to recovery. Tachycardia was finally controlled with flecainide while the patient was on left ventricular assist device support. The device was successfully explanted after 28 days of support. The temporary use of a left ventricular assist device was necessary to maintain a good hemodynamic status during the treatment of pharmacological refractory tachycardia, and it allowed a successful bridge to recovery. PMID- 16039172 TI - Quality, safety, and transparency: a rising tide floats all boats. PMID- 16039173 TI - What is the future of mortality prediction models in heart valve surgery? PMID- 16039174 TI - Should surgical errors always be disclosed to the patient? PMID- 16039175 TI - Radiation therapy potentiates effective oncolytic viral therapy in the treatment of lung cancer. AB - BACKGROUND: Replication-competent oncolytic herpes simplex viruses with deletion of the gamma(1)34.5 gene preferentially replicate in and kill malignant cells. The gamma(1)34.5 gene codes for ICP 34.5, a protein that enhances viral replication, and is homologous to growth arrest and DNA damage protein 34 (GADD34), a radiation-inducible DNA repair gene. We hypothesized that radiation therapy may potentiate efficacy of oncolytic viral therapy by upregulating GADD34 and promoting viral replication. METHODS: The A549 and H1299 lung cancer cell lines were infected with NV1066, an oncolytic herpes simplex virus, at multiplicities of infection (number of viral particles per tumor cell) of 0.1 to 0.5 in vitro with radiation (2 to 10 Gy) or without radiation. Viral replication was determined by plaque assay, cell-to-cell spread was determined by flow cytometry, cell kill was determined by lactate dehydrogenase assay, and GADD34 induction was determined by real-time reverse transcription-polymerase chain reaction and Western blot method. Evidence of synergistic cytotoxicity dependence with GADD34 induction is further confirmed by small inhibitory RNA inhibition of GADD34 expression. RESULTS: Using both the isobologram method and combination index method of Chou and Talalay, significant synergism was demonstrated between radiation therapy and NV1066 both in vitro and in vivo. As a result of such synergism, a dose reduction for each agent (2- to 6,000-fold) can be accomplished for a wide range of therapeutic effect levels without sacrificing tumor cell kill. This effect is correlated with increased GADD34 expression and inhibited by transfection of small inhibitory RNA directed against GADD34. CONCLUSIONS: These data provide the cellular basis for the clinical investigation of combined use of radiation therapy with oncolytic herpes simplex virus therapy in the treatment of lung cancer to achieve synergistic efficacy while minimizing dosage and toxicity. PMID- 16039176 TI - Low morbidity and mortality for bronchoplastic procedures with and without induction therapy. AB - BACKGROUND: The safety of bronchoplastic procedures after induction chemoradiotherapy is uncertain. This study examines short- and long-term outcomes after bronchoplastic procedures with and without induction therapy. METHODS: Between January 1997 and September 2004, more than 1,300 anatomic pulmonary resections for cancer were performed at a single institution. Of these, 73 patients required either sleeve lobectomy (57) or bronchoplasty (16), and were retrospectively analyzed. Nineteen patients (26%) received induction therapy; 15 received chemotherapy and radiation therapy and 4 received chemotherapy alone. Fifty-four patients underwent the bronchoplastic procedure without induction therapy. Mortality and early and late morbidity were analyzed. RESULTS: Mean follow-up was 25 months. Histology was nonsmall cell cancer in 62 (85%), carcinoid in 8 (11%), and renal cell cancer, schwannoma, and mucoepidermoid cancer in 1 patient each. There were 2 (2.7%) 30-day deaths, both in the group not receiving induction therapy. Of the surviving 71 patients, 70 had functional reconstructions at last follow-up. The overall 30-day complication rate was 30% (19 of 54) in patients not receiving induction therapy (no bronchopleural fistulas) and 42% (8 of 19) occurring in those receiving induction therapy (1 bronchopleural fistula). The long-term complication rate was 20% (11 of 54) among patients not receiving induction therapy and 5% (1 of 19) among those receiving induction therapy (completion pneumonectomy). There were no bronchovascular complications. Interventional bronchoscopy was required in 7 patients not receiving induction therapy, and was required in none of the patients receiving induction therapy. CONCLUSIONS: Anatomic pulmonary resections utilizing bronchoplastic techniques can be performed with low morbidity and mortality rates even after induction therapy. PMID- 16039177 TI - Preoperative chemotherapy does not increase complications after nonsmall cell lung cancer resection. AB - BACKGROUND: Neoadjuvant chemotherapy before resection of nonsmall cell lung cancer seems to increase survival, mainly in the early stage. Risks of postoperative complications after chemotherapy and surgery remain controversial. Here we review our experience with patients treated in one thoracic surgery center. METHODS: Patients undergoing resection for nonsmall cell lung cancer after induction chemotherapy between January 1993 and March 2002 were reviewed. Data collected included age, sex, preoperative forced expiratory volume in 1 second (FEV1), hemoglobin, and arterial oxygen pressure tension (PaO2), postoperative complications, and global survival. The main objectives were postoperative mortality and morbidity. Postoperative mortality and morbidity were defined as complications or deaths occurring within 30 days after surgery. Predictive morbidity factors were identified by univariate and multivariate analysis and overall survival by the Kaplan-Meier method. RESULTS: In all, 114 patients were reviewed. Different induction chemotherapies were used, mainly cisplatin with vinorelbine or gemicitabine. Postoperative mortality was 2 of 114, 1 of 27 after pneumonectomy, and there were no deaths after lobectomy. Complications occurred in 29% of patients (33 of 114), usually infectious pneumonia and anemia requiring transfusion. Preoperative FEV1, hemoglobin, and PaO2 are not associated with morbidity in univariate or multivariate analysis. CONCLUSIONS: Preoperative chemotherapy does not increase postoperative mortality and morbidity after nonsmall cell lung cancer surgery, performed exclusively by thoracic surgeons. PMID- 16039178 TI - Relevance of lymph node micrometastases in radically resected endobronchial carcinoid tumors. AB - BACKGROUND: Endobronchial carcinoids may recur even if radically resected. This retrospective study investigates the clinical and prognostic relevance of lymph node micrometastasis in these neoplasms. METHODS: Fifty-five patients underwent standard anatomic resection (lobectomy or pneumonectomy) with systematic routine (hilar and mediastinal) lymphadenectomy. After an evaluation of conventional prognostic factors, we reanalyzed lymph nodes of pN0 patients for micrometastasis using immunohistochemistry (anti-cytokeratin AE1/AE3 and anti-chromogranin A antibodies). RESULTS: We performed 9 pneumonectomies, 41 lobectomies, and 5 bilobectomies. Histologic examination showed 47 (85%) typical carcinoid and 8 (14.5%) atypical. Twelve patients were pN1 (8 typical, 4 atypical); after reanalysis another 8 revealed micrometastasis, 6 N1 micrometastasis (5 typical, 1 atypical), and 2 N2 micrometastasis (2 atypical), increasing subjects with nodal involvement (pN1 and N micrometastasis) from 12 (21.8%) to 20 (36.4%; p = 0.01). Micrometastases were more frequent in atypical carcinoids (p = 0.002). Local recurrence developed in 3 (5.4%) patients: 2 pN1 (1 typical, 1 atypical) and 1 N1 micrometastasis (1 typical). Distant relapse occurred in 2 (3.6%) patients, both N2 micrometastasis (2 atypical). After reanalysis, recurrence rate in patients with nodal disease increased from 16.7% to 25% (p = 0.01). All patients with recurrence died: all had pN1 or N micrometastasis. No patient confirmed as N0 had recurrence. Only histologic pattern and node status significantly influenced disease-free (p = 0.002 and p = 0.05) and overall survivals (p = 0.02 and p < 0.001), respectively. Micrometastasis worsen both disease-free (p < 0.0001) and overall (p < 0.001) survival rates at 5 and 10 years. CONCLUSIONS: Routine systematic lymphadenectomy with immunohistochemical detection of lymph node micrometastasis contributes to identification of a larger population at risk with a higher recurrence rate, allowing a more accurate staging of endobronchial carcinoids. PMID- 16039179 TI - Invited commentary. PMID- 16039180 TI - Outcomes of tracheobronchial stents in patients with malignant airway disease. AB - BACKGROUND: Malignant central airway obstruction is difficult to manage and is associated with poor outcome. We sought to identify the short (< 30 days) and intermediate (> 30 days) benefits and risks of tracheobronchial stents in patients with malignant airway disease. METHODS: Two hundred and twenty-five tracheobronchial stents were placed in 172 patients for benign (n = 32) and malignant (n = 140) disease from January 1, 1997, to May 31, 2003. The records of the patients with malignant disease were retrospectively analyzed to determine complication rate, reintervention rate, and survival. The malignant diagnoses included nonsmall cell cancer, small cell cancer, esophageal cancer, and metastatic disease. RESULTS: There were 172 stents placed in 140 patients with malignant disease, with no intraoperative mortality. The mean follow-up period was 142 +/- 12 days. There were 23 complications, including tumor ingrowth (n = 9), excessive granulation tissue (n = 7), stent migration (n = 5), and restenosis (n = 2). Five of the complications occurred during the short-term period (< 30 days) with the remaining complications (n = 18) occurring after 30 days. The complications required interventions including laser debridement (n = 14), dilation (n = 4), and stent removal (n = 5). CONCLUSIONS: Tracheobronchial stents offer minimally invasive palliative therapy for patients with unresectable malignant central airway obstruction. The benefit of airway stents is particularly seen in the short-term period where they provide symptomatic improvement and have low complication risk. The major impediment is excessive granulation tissue and tumor ingrowth, which occur primarily after 30 days. PMID- 16039181 TI - Preoperative detection of pleural adhesions by chest ultrasonography. AB - BACKGROUND: The presence of pleural adhesions may render video-assisted thoracoscopic surgery difficult or impossible. The aim of this study was to assess the value of chest ultrasonography in the detection of pleural adhesions prior to thoracotomy. METHODS: Between October 2001 and September 2002, 42 consecutive patients undergoing thoracotomies (including video-assisted thoracic surgery) were evaluated with chest ultrasonography. These patients underwent a preoperative ultrasonic examination of the chest wall using a 7-MHz linear ultrasound probe at 7 points along the chest wall. We measured the movement of the visceral pleural slide. RESULTS: When restricted viscera sliding was defined as less than 1 cm of excursion at the upper thoracic wall during exaggerated respirations, ultrasonography demonstrated a sensitivity of 63.6%, a specificity of 79.4%, a negative predictive value of 87.7%, a positive predictive value of 50.0%, and an overall accuracy of 75.6%. When restricted viscera sliding was defined as less than 2 cm of excursion at the lower thoracic wall during exaggerated respirations, ultrasonography demonstrated a sensitivity of 81.5%, a specificity of 81.0%, a negative predictive value of 96.0%, a positive predictive value of 44.0%, and an overall accuracy of 81.0%. CONCLUSIONS: Chest ultrasonography is moderately accurate in detecting the presence and location of pleural adhesions. Use of preoperative chest sonographic findings to plan trocar placement and to determine the need for an open approach is valuable in helping prevent visceral injury and facilitating video-assisted thoracoscopic surgery. PMID- 16039182 TI - Intrathoracic gastric emptying of solid food after esophagectomy for esophageal cancer. AB - BACKGROUND: Information on the function of the intrathoracic stomach after esophageal resection for esophageal cancer is limited. This study evaluated the factors affecting intrathoracic gastric emptying of solid food in patients who had undergone this surgery. METHODS: Between February 2003 and August 2003, intrathoracic gastric emptying of solid food was evaluated by radioisotope in 56 of the patients who underwent esophageal replacement surgery with the stomach for esophageal cancer. The 50% gastric emptying time was categorized into three ranges: over 180 minutes was defined as delayed, within 180 minutes as intermediate, and when all the radioisotopes were dumped into the jejunum as rapid. The factors affecting the gastric emptying were analyzed. RESULTS: In 6 out of 56 (10.7%) patients the radioisotope removal was too rapid to check the gastric-emptying time, and hence the analysis was performed in 50 patients. Twenty-one (37.5%) patients showed significantly delayed gastric emptying. The 50% gastric emptying time calculated by an exponential fitting method was 422 and 109 minutes in the delayed and intermediate groups, respectively. Age, the method of gastric drainage, and the stomach shape used did not affect the intrathoracic gastric emptying. Only the follow-up interval after surgery was a significant factor on gastric emptying (p = 0.024). The intrathoracic gastric emptying of solid food was more and more increased over time after surgery by regression analysis (p = 0.03). CONCLUSIONS: The use of the intrathoracic stomach as an esophageal substitute had a significant effect on gastric emptying, with intrathoracic gastric emptying of solid foods immediately after esophagectomy being markedly prolonged in about 50% of patients. However, the intrathoracic gastric emptying significantly increased over time. Research is required into improving intrathoracic gastric emptying in the early period after esophagectomy. PMID- 16039183 TI - Invited commentary. PMID- 16039184 TI - Incidence and management of chyle leakage after esophagectomy. AB - BACKGROUND: Postoperative chyle leakage is a rare but well-recognized complication after esophageal surgery. The aim of this study was to identify its incidence and potentially predisposing factors and to assess the consequences and management. METHODS: A consecutive series of 536 patients who underwent esophagectomy for malignant disease of the esophagus or gastroesophageal junction was reviewed. RESULTS: There were 20 patients (3.7%) with chyle leakage. After transthoracic esophagectomy the risk for the development of chyle leakage was higher than after transhiatal resection (p = 0.006). Chyle leakage was associated with more positive nodes (p = 0.041). Patients with chyle leakage had significantly more pulmonary complications (p < 0.001) and longer intensive care unit (p = 0.015) and hospital stays (p = 0.001). No patient with chyle leakage died. Conservative management, consisting of no enteral feeding and total parenteral nutrition, was instituted in all patients, but was abandoned in 4 patients (20%) because of persistence of high chyle output through the chest tube. In contrast to patients who were successfully treated with conservative measures, patients who eventually needed a reoperation had a drain output of more than 2 L on the day conservative therapy was started and 1 and 2 days later. CONCLUSIONS: Chyle leakage is seen more often in patients who undergo transthoracic esophagectomy and in patients who have more positive nodes. Patients with chyle leakage have more pulmonary complications. Conservative therapy is often successful, but operative therapy should be seriously considered in patients with a persistently high daily output of more than 2 L after 2 days of optimal conservative therapy. PMID- 16039185 TI - Thoracoscopic sympathectomy for axillary hyperhidrosis: the influence of T4. AB - BACKGROUND: Recent data suggest that severe compensatory sweating after sympathectomy for hyperhidrosis is more common than previously reported. In particular, T2-T4 sympathectomy for axillary hyperhidrosis leads to significantly more disabling sweating compared with T2-T3 sympathectomy for palmar hyperhidrosis. However, it is not known whether this is a result of the additional transection of the T4 segment or if patients with primary axillary hyperhidrosis are more prone to experience disabling compensatory sweating. METHODS: A follow-up study by questionnaire was made of 100 consecutive patients who underwent thoracoscopic sympathectomy for axillary hyperhidrosis at two university hospitals. Patients underwent T2-T3 sympathectomy (n = 35) or T2-T4 sympathectomy (n = 65) depending on the surgeon's preference. RESULTS: The questionnaire was returned by 91% of patients after a median of 31 months. Compensatory sweating occurred in 90% of patients and was so severe in 61% that they often had to change clothes during the day. There were no significant differences in occurrence or severity of compensatory sweating between the two extents of sympathectomy. Surgical outcome, however, was significantly better after T2-T4 sympathectomy. CONCLUSIONS: In contrast with previous reports, the incidence of compensatory sweating was not significantly related to the extent of sympathectomy for axillary hyperhidrosis. This result suggests that patients with primary axillary hyperhidrosis are more prone to experience compensatory sweating. Although the majority of patients with axillary hyperhidrosis were satisfied after thoracoscopic sympathectomy, many regret the operation. Patients should undergo surgery only if medical treatments fail; and provided there is an indication, we recommend T2-T4 sympathectomy. PMID- 16039186 TI - Health-related quality of life after thoracoscopic sympathectomy for palmar hyperhidrosis. AB - BACKGROUND: Palmar hyperhidrosis is a benign functional disorder regarded as a psychological and social handicap. Improvement of the quality of life is a major goal of treatment. However, little attention has been given to quality of life after thoracoscopic sympathectomy, which is the first line of treatment for palmar hyperhidrosis. This study investigated the impact of thoracoscopic sympathectomy on subjective health-related quality of life (HRQoL) and psychological properties. METHODS: Forty patients who underwent thoracoscopic sympathectomy were followed up for 6 months. The HRQoL measures were the Medical Outcomes Study Short Form 36 (SF-36), the Spielberger State Trait Anxiety Inventory (STAI), and the Zung Self-Rating Depression Scale (SDS). Patients were administered these questionnaires before procedure and then again at 1, 3, and 6 months after sympathectomy. RESULTS: A comparison between the current sample and Japanese normative data for the SF-36 showed mild impairment of HRQoL before sympathectomy. However, it also showed significant improvement of the social functioning domain after sympathectomy. While there was worsening of the bodily pain and role physical domains 1 month after sympathectomy, both domains recovered in 3 months. The results of STAI showed significant improvement of both trait and state anxiety after sympathectomy. However, the results of SDS showed patients remained neurotic. CONCLUSIONS: This study is the first to show the pattern of impairment in health status and therapeutic impact in palmar hyperhidrosis patients. Hyperhidrosis is associated with impaired HRQoL. It was also demonstrated that thoracoscopic sympathectomy is safe, minimally invasive, and improves HRQoL, even if compensatory hyperhidrosis occurs. PMID- 16039187 TI - Oximetry-derived perfusion index for intraoperative identification of successful thoracic sympathectomy. AB - BACKGROUND: Endoscopic thoracic sympathectomy treats patients with hyperhidrosis. Laser Doppler flow and changes in palmar temperature have been advocated for intraoperative monitoring. The pulse oximetry-derived perfusion index (PI) is used to quantify pulsatile blood flow at the pulse oximeter. Upper limb sympathectomy is associated with increased flow to the ipsilateral extremity; thus we postulate that it will increase PI. We evaluated changes in intraoperative PI as a monitor of successful thoracic sympathectomy. METHODS: After institutional review board approval and informed consent, 10 adult patients undergoing bilateral endoscopic thoracic sympathectomy under general anesthesia were studied. Finger pulse-oximetry probes were placed on each hand, and reference probes on each earlobe. Hemodynamic variables and PI were continuously monitored. Data were collected immediately before and every minute after sympathectomy for 5 minutes. Anesthetic management remained constant throughout. A successful sympathectomy was defined by a twofold increase in PI on the ipsilateral arm. Data were analyzed with analysis of variance and Student's t tests; a p < 0.05 was considered significant. RESULTS: Baseline oximetric waveforms were adequate in all subjects. Right sympathectomy was associated with a 372% increase in PI (p < 0.0001), and left sympathectomy with a 316% increase in PI (p < 0.029). This occurred as early as 1 minute after transection of the sympathetic chain. The PI in the reference probes as well as the hemodynamics remained constant. All patients had postoperative resolution of their hyperhidrosis symptoms. CONCLUSIONS: In patients with hyperhidrosis of the upper extremity, the intraoperative PI derived from pulse oximetry is an additional indicator of successful thoracic sympathectomy. PMID- 16039188 TI - Validation and refinement of mortality risk models for heart valve surgery. AB - BACKGROUND: The Northern New England Cardiovascular Disease Study Group (NNE) recently published risk models for hospital mortality after heart valve surgery. The Providence Health System Cardiovascular Study Group (PHS) has been collecting similar heart valve data for 8 years, providing an ideal opportunity to both validate the NNE risk models and attempt to produce an improved model, by using some different modeling techniques. METHODS: From 1997 to 2004, 3,324 patients aged 30 to 95 years underwent aortic valve replacement (AVR), and 1,596 underwent mitral valve replacement or repair (MVRR) at one of nine PHS medical centers. We used area under the receiver operating characteristic curve (c-index) to measure model discrimination, and Hosmer-Lemeshow statistic (H-L) to measure calibration. We modified the NNE models by ungrouping continuous variables, seeking optimal transformations of continuous variables, and imputing missing values by multiple regression. RESULTS: The prevalence and the lethality of risk factors were similar in PHS and NNE patients. The NNE models fit PHS patients well: c-index (95% confidence interval) = 0.75 (0.70 to 0.80) for AVR and 0.81 (0.76 to 0.86) for MVRR; H-L = 3.95 (p = 0.861) for AVR and 7.10 (p = 0.526) for MVRR. A single PHS model performed slightly better for both positions: c-index = 0.79 (0.75 to 0.83) for AVR and 0.84 (0.80 to 0.88) for MVRR; H-L = 2.75 (p = 0.949) for AVR and 12.21 (p = 0.142) for MVRR. CONCLUSIONS: The NNE models for aortic and mitral valve surgery were successfully validated using PHS patients. Using some different statistical approaches to modeling, we produced a new, unified model for both positions. PMID- 16039189 TI - Ten-year outcome after aortic valve replacement with the freestyle stentless bioprosthesis. AB - BACKGROUND: Stentless aortic bioprostheses offer excellent hemodynamics and potentially improved durability compared with other bioprostheses. The present report describes the clinical and hemodynamic outcomes for the Freestyle aortic root bioprosthesis in a large, multicenter cohort prospectively followed up for 10 years. METHODS: A total of 725 patients at 8 centers in North America (668 [92%] aged more than 60 years) were followed up prospectively after aortic valve replacement with the Freestyle stentless bioprosthesis. Implant technique was subcoronary in 509, total root in 178, and root inclusion in 38. Follow-up was 4,488 patient-years (mean 6.2 years/patient). RESULTS: For subcoronary, full root, and root inclusion groups, 10-year actuarial freedom from structural valve deterioration was 97.0% +/- 2.2%, 96.0% +/- 4.5%, and 90.9% +/- 11.2%, respectively; and actuarial freedom from reoperation was 91.7% +/- 3.5%, 92.3% +/ 6.0%, and 92.0% +/- 10.7%, respectively. Mean pressure gradient at 10 years was 8.9 +/- 7.9 mm Hg for subcoronary, 7.0 +/- 4.1 mm Hg for full root, and 10.0 +/- 11.1 mm Hg for root inclusion groups; effective orifice area was 1.6 +/- 0.5 cm2, 1.6 +/- 0.6 cm2, and 1.7 +/- 0.5 cm2, respectively. Fredom from moderate or more aortic regurgitation at 10 years was good for all three implant groups, but slightly higher for full root (97.7% +/- 1.6%) compared with subcoronary (87.2% +/- 2.8%) patients (p < 0.005). CONCLUSIONS: The Freestyle stentless aortic root bioprosthesis is a versatile option for aortic valve replacement. Measures of clinical outcomes and prosthesis durability remain excellent through 10 years. PMID- 16039190 TI - Aortic valve replacement with the pulmonary autograft: mid-term results. AB - BACKGROUND: The purpose of this study is to assess the mid-term results of aortic valve replacement with the pulmonary autograft. METHODS: From October 1993 through September 2003, 153 patients with aortic valve disease (81 rheumatic and 72 non-rheumatic), with a mean age of 28 +/- 14.2 years underwent the Ross procedure with root replacement technique and right ventricular outflow tract reconstruction using a homograft. Associated procedures included mitral valve repair (n = 19), open mitral commissurotomy (n = 15), tricuspid valve repair (n = 2), homograft mitral valve replacement (n = 2), and subaortic membrane resection (n = 1). RESULTS: Early mortality was 6.5% (10 patients). Mean follow-up was 77 +/- 42 months (range, 7 to 132 months; median, 90 months). One hundred, twenty one survivors (84.6%) had no significant aortic regurgitation. Reoperation was required in 10 patients for autograft dysfunction alone (n = 3), infective endocarditis (n = 2), autograft dysfunction with failed mitral valve repair (n = 3), and failed mitral valve repair alone (n = 2). No reoperations were required for the pulmonary homograft. There were 8 late deaths. Actuarial and reoperation free survival at 90 months were 91.% +/- 3.5%, 95.3% +/- 2.7%, in non-rheumatics and 86.1 +/- 3.9%, 90.5 +/- 3.7% in rheumatics, respectively. Freedom from significant aortic stenosis or regurgitation was 91.5 +/- 2.8% in non-rheumatics and 80.6 +/- 4.8% in rheumatics. Event-free survival was 86.2 +/- 4.9% in non rheumatics and only 68.9 +/- 5.3% in rheumatics. CONCLUSIONS: The Ross procedure is not recommended for young patients (< 30 years) with rheumatic heart disease. It provides satisfactory hemodynamic and clinical results in properly selected patients. Important autograft dilatation was not observed in our patients. PMID- 16039191 TI - Invited commentary. PMID- 16039192 TI - Long-term results after aortic valve replacement with the Bravo 400 stentless xenograft. AB - BACKGROUND: This study was undertaken to evaluate the long-term clinical and echocardiographic outcome after aortic valve replacement with the Bravo Cardiovascular Model 400 stentless xenograft. METHODS: Between February 1992 and January 1994, 67 patients underwent aortic valve replacement with the Bravo 400 bioprosthesis. The valvular pathology was aortic stenosis in 36 patients (53.7%), aortic insufficiency in 17 patients (25.4%), and mixed lesion in 14 patients (20.9%). Mean follow-up time was 9.8 +/- 2.73 years and median follow-up time was 11 years. Cumulative follow-up time was 659 patients-years and was 94% complete. RESULTS: No early deaths were observed. Overall survival estimates at 11 years were 74.71% +/- 5.47%. The actuarial freedom from valve-related death at 11 years was 91.04% +/- 3.84%; from cardiac-related death at 11 years it was 87.95% +/- 4.29%; and from noncardiac death at 11 years it was 85.14% +/- 4.58%. Eleven-year Kaplan-Meier survival of patients younger than 65 years was 90.91% +/- 6.13% versus 66.08% +/- 7.38% for older patients (p = 0.0307, log-rank test). The actuarial freedom from all valve-related morbidity and mortality at 11 years was 80.3% +/- 5.4%. The mean transvalvular gradient decreased significantly after aortic valve replacement with a corresponding increase in effective orifice area. Left ventricular mass index at 10-year follow-up was 68.5% of the preoperative value. CONCLUSIONS: The Bravo Cardiovascular Model 400 stentless xenograft has provided good clinical and hemodynamic results up until 11 years of follow-up. PMID- 16039193 TI - High-risk mitral valve surgery: perioperative hemodynamic optimization with nesiritide (BNP). AB - BACKGROUND: Nesiritide is a recombinant brain-type natriuretic peptide (BNP), which decreases pulmonary arterial (PA) pressures and myocardial oxygen consumption while increasing coronary flow and urine output. Mitral valve (MV) surgery in patients with severe mitral regurgitation (MR), impaired left ventricular function, and pulmonary hypertension is associated with a high operative mortality. We hypothesized that the perioperative use of Nesiritide is safe, and may improve surgical outcomes. METHODS: From May 2003 to August 2004, 14 patients (11 male, 3 female; mean age, 64 years [23-87 years]; mean systolic PA, 63 mm Hg [48-94 mm Hg]; mean ejection fraction, 36% [10-50%]), undergoing MV surgery (10 repairs, 2 replacements, and 2 rereplacements) for severe MR, were treated for a median of 24 hours (13-55 hours) preoperatively with intravenous Nesiritide. Expected mortality by EuroSCORE was 26% (7.8-59%) (5 reoperations). Concomitant procedures included tricuspid valve repair (n = 7), coronary artery bypass grafting (n = 5), and left atrial maze procedure (n = 3). Eleven patients received Nesiritide postoperatively during a mean duration of 22 hours (2-80 hours). RESULTS: Operative mortality was 0%. Prior to surgery after BNP treatment, mean systolic PA pressure dropped to 39 mm Hg (p = 0.0003), pulmonary capillary wedge pressure to 15 mm Hg (p = 0.001), central venous pressure to 6 mm Hg (p = 0.002), and weight by 3.7 kg (p = 0.006). Postoperative median ventilation time was 14 hours (4-48 hours). All other major hemodynamic parameters (systemic blood pressure, heart rate, and cardiac output) remained constant. The treatment was well-tolerated in all patients. CONCLUSIONS: Perioperative use of Nesiritide is safe, and may contribute to improved early outcomes in high-risk patients undergoing MV surgery. This may be due to improved ventricular loading conditions (decreased PA pressures, more effective diuresis) and/or a direct myocardial effect of BNP. Further prospective evaluation of the role of BNP in cardiac surgery is warranted. PMID- 16039194 TI - A novel approach to tricuspid valve replacement: the upside down stentless aortic bioprosthesis. AB - BACKGROUND: Tricuspid valve replacement (TVR) is a rarely needed operation. Choices between mechanical and biological prosthesis still generate controversy. We present our initial clinical experience with a stentless aortic root placed inverted in the tricuspid annulus. METHODS: Between August 2000 and September 2003, TVR for severe tricuspid insufficiency was performed in 8 patients. Indications were infective endocarditis (7) and iatrogenic damage (1). Mean age was 42.2 years old (20 to 58 years old). Five patients were male and 3 patients had a concomitant procedure (mitral valvuloplasty, coronary bypass grafting, and aortic valve replacement). A stentless aortic root, size 27 mm (n = 5) or 29 mm (n = 3) was placed inverted in the tricuspid position after the valsalva sinuses were scalloped. Interrupted 4-0 polypropylene sutures were used between the tricuspid valve annulus and the sewing ring. The struts equivalent on the stentless valve were anchored to the septal, anterior and posteroinferior wall of the right ventricle using 5-0 PTFE pledgeted sutures. RESULTS: Hospital survival was 100% and mean hospital stay was 12.5 days (3 to 18 days). Intraoperative and follow-up echocardiograms revealed no stenosis or insufficiency. Mean follow-up was 17.2 months (1-38 months). There were 3 late deaths due to continued IV drug use (n = 2) and end-stage renal failure (n = 1). CONCLUSIONS: This is a novel surgical alternative for a very high risk population. Potential advantages over current options include minimization of blood contact with nonbiological surfaces, preservation of annular motion, freedom from anticoagulation, and a theoretical lower rate of calcification. PMID- 16039195 TI - Metabolic and hemodynamic effects of high-dose insulin treatment in aortic valve and coronary surgery. AB - BACKGROUND: Glucose and insulin have been used as an adjuvant therapy in cardiac surgery because of their potentially beneficial effects on myocardial metabolism and contractile function. This study evaluated the effects of high-dose insulin on systemic metabolism and hemodynamics after combined heart surgery. METHODS: Forty elective patients scheduled for combined aortic valve replacement and coronary artery bypass surgery were randomly assigned to receive either high-dose insulin treatment (short-acting insulin 1 IU.kg(-1).h(-1) with 30% glucose 1.5 mL.kg(-1).h(-1) administered separately) or control treatment (saline). The blood glucose levels were maintained within a targeted range by adjusting the rate of glucose infusion in the treatment group and by short-acting insulin bolus doses in the control group. RESULTS: The lactate clearance was faster (p = 0.046), and the lactate levels (p = 0.016), blood glucose levels (p < 0.001), and free fatty acid levels (p < 0.001) were lower in the insulin group postoperatively. Besides, there was lesser need for dobutamine support (p = 0.013) and a trend toward better cardiac indices. Insulin treatment increased the respiratory quotient (p < 0.001), but there were no differences between the groups with regard to systemic oxygen consumption or energy expenditure measured by indirect calorimetry. The average glucose uptake in the insulin group was 7.1 g/kg in 24 hours (28 kcal.kg( 1).day(-1)). CONCLUSIONS: The high-dose insulin treatment was associated with lower blood glucose levels, better preserved myocardial contractile function, and less need for inotropic support, and hence led to lower lactate levels postoperatively. The protocol is safe, but requires strict control of blood glucose level. PMID- 16039196 TI - Early postoperative use of unfractionated heparin or enoxaparin is associated with increased surgical re-exploration for bleeding. AB - BACKGROUND: A variety of indications (eg, prosthetic heart valves, atrial fibrillation, etc.) exist for the use of unfractionated heparin (UFH) and enoxaparin (ENOX) in the early postoperative period following open-heart surgery. However, the overall postoperative risk for hemorrhage from the use of UFH and ENOX are not known. METHODS: From 1998 to 2001, 2,977 consecutive open-heart or valve surgery patients were retrospectively evaluated. Postoperatively, 2,037 received no UFH or ENOX, 579 received intravenous UFH, and 361 received ENOX. Baseline characteristics were collected, patients who required surgical re exploration for postoperative bleeding and time between surgery and re exploration were followed-up. RESULTS: Average patient ages were 64 +/- 13, 65 +/ 12, and 68 +/- 10 years receiving none, UFH (p < 0.01 vs none), and ENOX (p < 0.01 vs none; p < 0.01 vs UFH), respectively. Rates of surgical re-exploration were 2.7% for none, 7.8% for UFH, and 8.9% for ENOX (vs none, adjusted hazard ratio = 2.8; p < 0.001 for UFH; hazard ratio = 3.3; p < 0.001 for ENOX). Males were also at higher risk for re-exploration (hazard ratio = 1.4; p = 0.07). For those requiring re-exploration, the interval between surgery and first re exploration was prolonged (> 4 days) among those receiving ENOX (37.5%, odds ratio = 36.7; p = 0.001) and UFH (20.0%, odds ratio = 14.7; p = 0.01) compared with none (1.8%). Prolonged times with ENOX had a greater proportion of prolonged times than UFH (odds ratio = 2.5; p = 0.09). CONCLUSIONS: Early postoperative use of ENOX and UFH is associated with a significant increase in re-exploration for postoperative bleeding, often at a significantly delayed time period after the initial surgery. This delay was especially common with ENOX suggesting the need for prospective studies. PMID- 16039197 TI - Long-term results after 27 years of surgical treatment of acute type a aortic dissection. AB - BACKGROUND: This study investigates the determinants of long-term outcome and modalities of late death after surgical treatment of acute type A dissection. METHODS: Between 1974 and 2001, 315 consecutive patients were operated on for acute type A aortic dissection. Operative mortality was 22.9%. A series of 243 survivors of surgical treatment were followed up for as long as 27 years. Endpoints were death, cardiovascular reoperation, and neurologic events. Median follow-up was 4.5 years. Follow-up was 99.6% complete. RESULTS: Cumulative survival of discharged patients was 96.4% +/- 1.3%, 67.7% +/- 4.7%, and 39.4% +/- 12.0% at 1, 10, and 20 years, respectively. During follow-up, 47 patients died. Cause of death was cardiac failure in 7, hemorrhage due to rupture of the distal aorta in 7, stroke in 4, respiratory insufficiency in 4, sepsis in 3, malignancy in 2, and unknown in 20 patients. Multivariate analysis revealed advanced patient age and postoperative hemodialysis as perioperative indicators of late death (p < 0.05). Freedom from cardiovascular reoperation was 90.7% +/- 2.0% at 1 year, 60.9% +/- 5.1% at 10 years and 41.9% +/- 15.0% at 20 years. A total of 58 patients required 86 cardiovascular reoperations; aortic root or ascending aorta replacement was performed in 20, distal ascending aorta and arch replacement in 13, descending aorta replacement in 6, thoracoabdominal aorta replacement in 7, abdominal aorta replacement in 7, and miscellaneous reoperations in 6 patients. Multivariate analysis revealed male sex and left coronary artery dissection as significant determinants for late cardiovascular reintervention (p < 0.05). Cumulative incidence of stroke after 20 years was 3.8%. CONCLUSIONS: Acute type A dissection represents an emergency situation with acceptable long-term results for discharged survivors of surgical treatment. PMID- 16039198 TI - Invited commentary. PMID- 16039199 TI - Antimicrobial drug resistance in salmonella-infected aortic aneurysms. AB - BACKGROUND: Salmonella infection of the aorta and adjacent arteries is rare, but life-threatening. There is an increasing number of infections caused by antimicrobial drug resistant Salmonella. This study sought to assess the association between antimicrobial drug resistance and clinical outcomes of patients with Salmonella-infected aortic aneurysm. METHODS: Data were collected by retrospective chart review. Between October 1995 and October 2004, 34 patients with Salmonella-infected aortic aneurysm were included. Aneurysm-related deaths were defined as hospital deaths and late deaths due to prosthetic graft infection. Analysis was performed using the chi2 test, Fisher's exact test, and Mann-Whitney test. RESULTS: Nineteen patients had a suprarenal and 15 patients had an infrarenal aortic infection. The most common responsible pathogen was group C Salmonella (47%). Ciprofloxacin-resistant Salmonella infection occurred since March 2001 and the rate increased from 0 per 15 in the years before March 2001 to 5 per 19 in the years after March 2001 (p = 0.005 by Fisher's exact test). Among the 26 patients who had combined medical and surgical therapy, 4 died in the hospital and 4 died of late prosthetic graft infection 3 to 6 months after operation, whereas 4 of the 8 who had medical therapy alone died of aneurysm rupture during hospitalization. The actuarial survival rates by the Kaplan-Meier method were 64% at 6 months, 61% at 1 year, and 56% at 5 years. The risk factors for aneurysm-related death were old age (78.5 +/- 9.7 years vs 63.5 +/- 11.4 years; p < 0.001) and ciprofloxacin-resistant Salmonella infection (4 of 5 vs 8 of 29; p = 0.042). CONCLUSIONS: There was an increased mortality associated with ciprofloxacin resistance in infected aortic aneurysms with Salmonella. With an increasing incidence of ciprofloxacin resistant Salmonella, third generation cephalosporin is the antibiotic of choice for Salmonella infected aneurysm. PMID- 16039200 TI - Absent long-term benefit of patch versus linear reconstruction in left ventricular aneurysm surgery. AB - BACKGROUND: Endoventricular patch reconstruction of the left ventricle is considered the gold standard in surgery for left ventricular aneurysms, because of improved preservation of ventricular geometry. However, the superiority over conventional linear closure has not been demonstrated, as assessed by the long term outcome. METHODS: Two hundred patients (66%) underwent linear closure (group L) and 105 patients (34%) had endoventricular patch reconstruction (group D) using the Dor technique. Linear closure has been performed since 1974 and from 1985 on the Dor technique has been applied as an alternative procedure. Both patient groups differed regarding age, sex distribution, site of infarction, and indication for surgery. Prior to the operation, 71% of the patients were in New York Heart Association (NYHA) class III or IV and mean ejection fraction was 34% +/- 12%. Follow-up extends up to 25 years, with a cumulative total of 2,605 patient years. RESULTS: Early mortality was 6.5% in group L vs 5.7% in group D (not significant [NS]). Actuarial survival after 10 years was 56 +/- 3.2%, with no difference between groups. Freedom from reoperation after 10 years was 95.6% in group L vs 95.2% in group D (NS). Preoperative risk factors for late mortality were age, left ventricular enddiastolic volume index and concomitant mitral valve surgery. The type of procedure and the date of operation had no influence on mortality. To date, 63% of the survivors are in NYHA class I and II. CONCLUSIONS: In regard to long-term survival, rate of reoperation, and postoperative NYHA functional class, no benefit could be demonstrated when linear closure was compared with ventricular patch reconstruction for LV aneurysm repair. Hence, the technique of ventricular reconstruction may not be as important as previously thought, and at least for small aneurysms the simple and time sparing technique of linear closure may still be considered. PMID- 16039201 TI - A multidisciplinary network to save the lives of severe, persistent cardiogenic shock patients. AB - BACKGROUND: Severe, persistent cardiogenic shock is associated with very high morbidity and lethality. We therefore tried to improve the outcome of these patients using a network between local hospitals and a specialized heart center that has facilities for transplantation and ventricular assist device (VAD) implantation. METHODS: Between 1998 and 2000, 50 patients, who were in cardiogenic shock (mean age 49 years, SD 14) despite high doses of at least three inotropic agents, were admitted to our specialized heart center. During the first 12 hours, we tried to stabilize the patient's condition, to reduce inotropic support agents, and to keep cardiac index above 2.4 L.min(-1).m(-2). According to this strategy, patients were allocated to medical treatment only, emergency VAD therapy, urgent VAD implantation, or heart transplantation. To assess variables potentially influencing patients' outcome, we performed univariate and multivariate analyses. RESULTS: Early in-hospital mortality was 36% and late mortality was 10%. Overall survival at 1, 6, and 12 months of follow-up was 70%, 56%, and 52%, respectively. Even when the heart transplant patients were excluded, overall 12-months survival after cardiogenic shock was still 50%. Higher age, and the absence of sinus rhythm were the only independent risk factors for 12-month mortality. CONCLUSIONS: Our data show that a network between cardiology departments of local hospitals and specialized heart centers can be life-saving for severe, persistent cardiogenic shock patients. PMID- 16039202 TI - CardioWest total artificial heart: Bad Oeynhausen experience. AB - BACKGROUND: The use of ventricular assist devices (VAD) has become a widely accepted therapeutic option. However, there are still limitations to the patient collective eligible for VAD placement, who might therefore benefit from the implantation of a total artificial heart. We present the first German single center experience with the CardioWest total artificial heart (TAH) (SynCardia Systems, Tucson, AZ) in 42 patients. METHODS: Between February 2001 and December 2003, 42 patients (37 men, 5 women, mean age 51 +/- 13 years) received a TAH at our Center. Their body surface area ranged between 1.5 and 2.4 (mean, 1.9 +/- 0.19) m2. All patients were in persistent cardiogenic shock in spite of maximum inotropic support and had numerous preoperative risk factors (intraaortic balloon pumping, mechanical ventilation, acute renal failure, previous cardiac surgery, recent cardiopulmonary resuscitation). RESULTS: Duration of support was 1 to 291 days. Eleven patients (26%) underwent successful transplantation; 9 of them could be discharged home. Twenty-two patients died under support, 21 of them from multiple organ failure and 1 patient from a technical problem. Nine patients are still on the device, 4 of them at home after the original CardioWest console was replaced by the Berlin Heart EXCOR driver (Berlin Heart, Berlin, Germany). Exceptional results were achieved in patients with cardiogenic shock after cardiac surgery or after acute myocardial infarction. CONCLUSIONS: Against the background of the extremely poor preoperative situation of our patients, the overall survival rate of 48% can be considered as favorable. A prospective, randomized study is planned to find out whether patients with idiopathic dilated or ischemic cardiomyopathy are more likely to benefit from a biventricular assist device or a total artificial heart. PMID- 16039203 TI - Mid-term results after thoracoscopic transmyocardial laser revascularization. AB - BACKGROUND: Transmyocardial revascularization is a surgical therapy for the relief of severe angina in patients who are not suitable candidates for coronary artery bypass graft surgery or percutaneous coronary interventions. Historically, surgical techniques employed a left thoracotomy with or without thoracoscopic assist for visualization. This study evaluated the feasibility and midterm outcomes after transmyocardial laser revascularization performed using a completely thoracoscopic, closed chest approach. METHODS: Patients (9 men [90%] and 1 woman [10%]) at a mean age of 66 +/- 10 years who were ineligible for coronary artery bypass graft surgery or percutaneous coronary intervention underwent sole therapy transmyocardial laser revascularization using a completely thoracoscopic surgical approach using a holmium:yttrium-aluminum-garnet laser system. Preoperatively, patients had a mean ejection fraction of 0.51 +/- 0.09 and a mean angina class of 3.7 +/- 0.5. RESULTS: A mean of 30 +/- 2.4 channels were created during mean laser and operative procedure times of 14 +/- 2.9 and 133 +/- 32 minutes, respectively. Patients were extubated at a mean of 7.6 +/- 12 hours and were discharged from the hospital at a mean of 5.4 +/- 3.4 days. There were no hospital deaths or major complications. At a mean of 8.4 +/- 5.5 months postoperatively, all patients survived and significant clinical improvement with a mean angina class of 1.3 +/- 0.5 (p < 0.001). CONCLUSIONS: A completely thoracoscopic surgical approach is feasible for sole therapy transmyocardial revascularization that affords improved visualization over a limited thoracotomy approach. Limited complications and significant clinical improvement after the procedure were observed. With minimal port manipulation, there is an opportunity for decreased postoperative pain; however, larger studies are warranted to verify this hypothesis. PMID- 16039204 TI - Radial artery grafts in women: utilization and results. AB - BACKGROUND: Despite a known survival benefit with the use of the left internal mammary artery, it is used less frequently in women when compared with men. This study evaluated the hypotheses that the radial artery graft is used less frequently in women compared with men, that the radial artery is smaller in women compared with men, and that the use of the radial artery influences operative mortality and long-term survival in women. METHODS: The use of a radial artery graft was evaluated in 2,633 patients who underwent isolated coronary artery bypass. Radial artery size and flow were compared in 207 patients who had intraoperative radial artery diameter and flow measurements. Propensity scoring was utilized to compare short- and long-term outcomes in a matched cohort of 588 women. RESULTS: Of 862 women (33%) who had isolated coronary artery bypass grafting, only 301 (35%) received a radial artery graft versus 44% of men (786 of 1,771, p < 0.001). Radial artery size and flow were significantly less in women. Operative mortality was not different between women with a radial artery graft and women without; however, 5-year survival was significantly better in women with a radial artery graft than in those without. CONCLUSIONS: Women received fewer radial artery grafts than men. Radial artery size and flow were significantly less in women than in men. Use of a radial artery graft did not influence operative mortality among women. However, 5-year survival among women who received a radial artery graft was significantly better than among women who did not. PMID- 16039205 TI - Invited commentary. PMID- 16039206 TI - Concomitant brachiocephalic and coronary artery disease: outcome and decision analysis. AB - BACKGROUND: In patients with coronary artery disease, concomitant brachiocephalic disease may affect outcome and influence decision making regarding operative staging, technique, and choice of conduit. METHODS: Eighty consecutive patients (mean age, 59.3 years; 60.0% male) with concomitant brachiocephalic and coronary artery disease were identified either before (group A, n = 48) or after (group B, n = 32) coronary artery bypass grafting. Patients who had symptomatic brachiocephalic and coronary artery disease before surgery underwent concomitant brachiocephalic reconstruction and coronary artery bypass grafting using either all-vein coronary conduits (n = 41) or vein-and-internal mammary artery conduits (n = 7). Patients who had coronary-subclavian steal syndrome after coronary artery bypass (group B, n = 32) underwent either surgical (n = 5) or endovascular (n = 27) brachiocephalic reconstruction only. RESULTS: All patients were asymptomatic after intervention. Operative mortality was 4.2% for group A and 3.1% for group B. The perioperative stroke rate was 2.1% for group A and 0% for group B. Actuarial 10-year freedom from specific events for group A was as follows: death 59.9 +/- 12.8%, brachiocephalic restenosis 100%, coronary subclavian steal syndrome 100%, myocardial infarction 83.5 +/- 10.5%, stroke 82.1 +/- 9.9%, redo coronary artery bypass grafting 95.8 +/- 4.1%, other vascular operation 82.2 +/- 8.9%, and adverse cardiac outcome (death, redo coronary artery bypass grafting, or myocardial infarction) 52.9% +/- 13.2% (for patients with all vein conduits) or 100% (for patients with vein-and-internal mammary artery conduits). At midterm follow-up (mean, 2.92 years), both the surgical and the endovascular treatment subgroups of group B had 100% brachiocephalic patency. CONCLUSIONS: Long-term results in a limited population support continued evaluation of concomitant brachiocephalic reconstruction and coronary artery bypass grafting with use of the internal mammary artery conduit in an attempt to improve late survival in patients with concomitant disease. The excellent midterm brachiocephalic patency after either surgical or endovascular treatment of patients with coronary-subclavian steal syndrome supports continued evaluation of both methods. PMID- 16039207 TI - Long-term survival after surgical revascularization for moderate ischemic mitral regurgitation. AB - BACKGROUND: We sought to characterize patient survival and degree of late mitral regurgitation (MR) in patients undergoing surgical revascularization with moderate ischemic MR. METHODS: We retrospectively reviewed 251 patients undergoing coronary artery bypass graft (CABG) surgery between 1991 and 2001 with 3+ ischemic MR, including 31 patients who had concomitant mitral annuloplasty. Univariate and multivariable testing was employed. RESULTS: Actuarial 1-, 5-, and 10-year survival was 84.0%, 67.5%, and 37.1% in the overall group of 251 patients. Independent predictors of long-term mortality were age 70 years or more (hazard ratio 2.50 [95% confidence interval 1.82 to 3.44]), prior myocardial infarction (3.99 [2.15 to 7.39]), unstable angina (2.27 [1.69 to 3.04]), chronic renal failure (4.87 [3.13 to 7.58]), atrial fibrillation (2.21 [1.65 to 2.96]), left internal mammary artery to left anterior descending artery graft (0.28 [0.18 to 0.43]), preoperative beta-blocker (0.43 [0.28 to 0.67]), ejection fraction (0.71/10% [0.64 to 0.80]), left atrium size (0.88/mm [0.84 to 0.92]), diffuse wall motion abnormalities (2.83 [1.77 to 4.55]), and mitral leaflet restriction (3.85 [2.46 to 5.99]). The model controlled for the performance of annuloplasty, which did not emerge as an independent predictor. Patients undergoing annuloplasty did have less mean late MR than those having CABG alone (p = 0.005). Overall, 57.8% of patients (63 of 109) with follow-up echocardiograms had improvement in grade of MR compared with baseline. In 54 of 95 patients (56.8%), intraoperative transesophageal echocardiography downgraded the degree of MR compared with the preoperative study. CONCLUSIONS: Patients with moderate ischemic MR undergoing CABG had relatively poor long-term survival, with significant differences when stratified according to preoperative characteristics. Performance of mitral annuloplasty reduced the degree of regurgitation but was not a predictor of long-term survival. Intraoperative transesophageal echocardiography frequently downgraded the degree of MR. PMID- 16039208 TI - Invited commentary. PMID- 16039209 TI - Total arterial myocardial revascularization using composite and sequential grafting with the off-pump technique. AB - BACKGROUND: Multiple arterial myocardial revascularizations are increasingly undertaken using off-pump techniques; however, various arterial grafting techniques are utilized. This study aimed to review the outcome of combining arterial composite and sequential grafting with off-pump techniques. METHODS: We retrospectively reviewed the records of 107 consecutive patients who underwent coronary bypass surgery with off-pump and arterial composite grafting techniques between April 2001 and March 2004. The left internal thoracic artery (LITA) was harvested in all patients, and the right internal thoracic artery (RITA), the radial artery (RA), and the gastroepiploic artery (GEA) were harvested in 69 patients, in 83 patients, and in 53 patients, respectively. Early postoperative angiograms were evaluated in 97 patients. RESULTS: There were 488 distal anastomoses, an average of 4.5 per patient. Forty-four in situ LITAs were used as LITA Y-composite grafts with a free RITA (n = 19), RA (n = 24), or free GEA (n = 1). Forty-three in situ RITAs were used as RITA-RA grafts (n = 42) or a RITA-GEA graft (n = 1). Twenty-one in situ GEAs were used as composite grafts with the RA (n = 17) or a free RITA (n = 4). There were no hospital deaths. The patency of the LITA was 100%, and that of the RA was 97.3%, while patencies of both in situ and free RITA and GEA were 100%. During the follow-up period (mean: 22.1 months), there were 3 late deaths, but none were cardiac related. CONCLUSIONS: Total arterial revascularization with composite and sequential grafting is a safe and effective technique in patients undergoing off-pump coronary artery bypass surgery. PMID- 16039210 TI - Hemostasis in off-pump compared to on-pump coronary artery bypass grafting: a prospective, randomized study. AB - BACKGROUND: We hypothesized that off-pump coronary artery bypass grafting has less impact on the hemostatic systems than on-pump surgery. METHODS: Thirty-one patients were randomized to on-pump or off-pump coronary artery bypass grafting. Factors of hemostasis as well as markers of endothelial activation were measured up to 24 hours after the operation: Fibrin D dimer, prothrombin fragment 1+2, alpha2-macroglobulin, protein C1 esterase inhibitor, fibronectin, and von Willebrand factor. Overall hemostasis potential, overall coagulation potential, and overall fibrinolysis potential were determined with a previously developed assay. We also measured platelet count before and after surgery. RESULTS: Fibrin D dimer and prothrombin fragment 1+2 concentrations were lower during surgery in the off-pump group (p < 0.001). Four hours after admission to the intensive care unit, these differences were eliminated. alpha2-macroglobulin, protein C1 esterase inhibitor, fibronectin, and von Willebrand factor concentrations did not differ between groups (p = 0.59, p = 0.28, p = 0.22, and p = 0.69). Protein C1 esterase inhibitor and von Willebrand factor concentrations increased over time (p < 0.001) in both groups. Overall hemostasis potential and overall coagulation potential increased over time (p < 0.001), while overall fibrinolysis potential decreased (p < 0.001) with no difference between groups (p = 0.69, p = 0.91). Platelet count decreased on the first postoperative day (p < 0.001), but increased from the first to the third postoperative day (p = 0.004) in both groups without any inter group difference (p = 0.82). CONCLUSIONS: There was a tendency toward less activation of coagulation and fibrinolysis in low-risk patients during elective off-pump coronary artery bypass surgery when compared with on-pump surgery. PMID- 16039211 TI - Prediction of graft flow impairment by intraoperative transit time flow measurement in off-pump coronary artery bypass using arterial grafts. AB - BACKGROUND: We assessed the validity of intraoperative transit time flow measurement (TTFM) in predicting graft flow abnormalities. METHODS: Intraoperative graft flow measurement using TTFM and early postoperative coronary angiography was performed in 58 patients who underwent total arterial off-pump coronary artery bypass. Five variables (flow pattern, mean flow, pulsatility index, insufficiency ratio, and fast Fourier transformation ratio) were measured and compared between 103 normal and 14 abnormal (occluded or competitive) grafts. RESULTS: The grafts anastomosed to the right coronary territories showed significantly less diastolic dominant pattern, lower mean flow and fast Fourier transformation ratio, and higher pulsatility index than grafts to the left coronary artery territories (p < 0.05). None of the abnormal grafts showed a diastolic dominant flow pattern. The abnormal grafts demonstrated significantly lower mean flow and fast Fourier transformation ratio and higher pulsatility index and insufficiency ratio than normal grafts (p < 0.05). When our criteria for detection of abnormal graft flow, [(1) systolic dominant or balanced pattern of the flow curve in the left coronary territories, systolic dominant pattern of the flow curve in the right coronary territories; (2) mean flow < 15 mL/min; (3) pulsatility index > 3 in the left coronary territories and > 5 in the right coronary territories; and (4) insufficiency ratio > 2%] were applied, the sensitivity and specificity of TTFM to detect the graft flow abnormality were 96.2% and 76.9%, respectively. CONCLUSIONS: Our data suggest that TTFM is a reliable intraoperative tool to predict graft flow impairment. PMID- 16039212 TI - Invited commentary. PMID- 16039213 TI - Intramyocardial left ventricle-to-coronary artery stent: a novel approach for the treatment of coronary artery disease. AB - BACKGROUND: The direct intramyocardial left ventricle-to-coronary artery stent may provide an optional minimally invasive technique for coronary artery bypass graft surgery. We seek to test whether blood flow and regional myocardial function improve with this stent in totally ischemic myocardium. METHODS: The stent device was implanted in 8 anesthetized dogs using an open chest approach, arteriotomy of the proximal left anterior descending coronary artery, and connection of the vessel to the left ventricular chamber. Regional coronary blood flow and myocardial function were monitored under three conditions: normal coronary flow (baseline), coronary ligation, and stent flow. RESULTS: Left anterior descending coronary ligation markedly reduced coronary artery blood flow and regional myocardial function. With flow solely from the stent, the blood flow pattern changed to one with high peak forward flow during systole compared with baseline (94.8 +/- 48.9 versus 56.8 +/- 21.1 mL/min; p < 0.05) and one with significant negative backflow during diastole compared with baseline (-37.4 +/- 23.1 versus 11.3 +/- 17.2 mL/min; p < 0.05). However, the resultant mean forward flow increased to approximately 50% of baseline compared with less than 5% of baseline after coronary ligation. Regional myocardial function diminished entirely after coronary ligation, but recovered to approximately 60% of baseline with the stent. Normal systemic hemodynamics and global ventricular contractile function were maintained with the stent. CONCLUSIONS: The left ventricle-to coronary artery stent is a simple and readily deployable device that allows the perfusion of epicardial vessels directly from the left ventricle and can provide significant blood flow to improve the performance of ischemic myocardium. It may provide an effective, alternative means of treating coronary artery disease when standard coronary artery bypass graft surgery is not suitable. PMID- 16039214 TI - Video-assisted thoracoscopic pericardial window for diagnosis and management of pericardial effusions. AB - BACKGROUND: Video-assisted thoracoscopy with the creation of a pericardial window is a noninvasive method of pericardial drainage. It also allows an excellent view of both the pleural cavity and pericardium and the precise selection of biopsy sites. We review our 3-year experience with this technique. METHODS: Between January 2001 and February 2004, 18 patients (10 men, 8 women; mean age 57 years) with echocardiographically diagnosed pericardial effusion underwent video assisted thoracoscopy at our center. Pericardial windows were created under general anesthesia and single-lung ventilation using 2 to 3 trocars. Mean operating time was 46 minutes. A right thoracic approach was used in 16 patients and a left thoracic approach in 2. RESULTS: Microbiology and virology cultures of the pericardial fluid were negative. Histologic findings were compatible with tuberculosis in 2 cases and granulocytic sarcoma, infiltrating breast carcinoma, and infiltrating nonsmall cell carcinoma in 1 case each. In the remaining patients, the histologic diagnosis was chronic or subacute nonspecific pericarditis. Talc pleurodesis was performed in 3 patients for concomitant malignant pleural effusion. In 4 patients, the pericardial effusion occurred secondary to cardiac surgery; 3 were receiving anticoagulants after valve replacement, and 1 had a heart transplant. There were no complications of the thoracoscopy technique. CONCLUSIONS: Video-assisted thoracoscopic fenestration is an effective technique for pericardial drainage and biopsy. Apart from its diagnostic value, it allows the physician to fashion a pleuropericardial window for effective drainage while avoiding the complications of classic surgical procedures. Concomitant pleural and pulmonary disorders may be managed simultaneously. PMID- 16039215 TI - Sivelestat reduces inflammatory mediators and preserves neutrophil deformability during simulated extracorporeal circulation. AB - BACKGROUND: Neutrophil is a major focus in efforts to ameliorate the systemic inflammatory response associated with cardiopulmonary bypass. Neutrophil elastase is a powerful proteolytic enzyme, and plays a pivotal role in the development of the inflammatory response. This study assesses the inhibitory effects of sivelestat, a highly specific neutrophil elastase inhibitor, on elastase levels, cytokine production, and the functional changes of neutrophils in a simulated extracorporeal circulation model. METHODS: Simulated recirculation was established by recirculating heparinized (3.75 U/mL) human blood for 120 minutes in an oxygenator and a roller pump circuit with and without 100 micromol/L of sivelestat (n = 7 for each group). Neutrophil elastase and interleukin-8 were measured with an enzyme immunoassay. Neutrophil deformability was evaluated by simulated microcapillaries. The neutrophil F-actin and the expression of CD11b and L-selectin were measured by flow cytometry. RESULTS: Sivelestat reduced both neutrophil elastase levels (p = 0.0006) and interleukin-8 production (p < 0.0001) at 120 minutes of recirculation. Sivelestat also significantly preserved neutrophil deformability (p = 0.017) and reduced F-actin expression (p = 0.0037). The drug did not modulate the changes of CD11b or L-selectin. CONCLUSIONS: This study suggests that specific elastase inhibition with sivelestat could be a feasible therapeutic strategy for patients undergoing cardiopulmonary bypass to attenuate neutrophil-derived inflammatory response and organ injuries. PMID- 16039216 TI - Incidence of deep sternal wound infections after tracheostomy in cardiac surgery patients. AB - BACKGROUND: Tracheostomy after median sternotomy is associated with an increased risk of deep sternal wound infection (DSWI). However, associated comorbidities in these patients make the exact risk of tracheostomy difficult to discern. Therefore, we performed a retrospective review to determine the risk of DSWI in our cardiac surgery patients who underwent postoperative tracheostomy. METHODS: Our cardiac surgery database was queried from January 1996 to July 2003. Perioperative morbidities were identified for all patients who underwent tracheostomy after cardiac surgery. Statistical analysis was performed using chi2 analysis and Fisher's exact test. RESULTS: During the study period, 16,277 cardiac procedures were performed through a median sternotomy. Tracheostomy was performed in 291 patients (1.8%). Deep sternal wound infections occurred in 0.8% of all patients and in 3.4% of patients (9 of 268) who underwent tracheostomy. Mean number of days to tracheostomy was 14.2 in the DSWI group and 15.8 in the non-DSWI group (p = 0.45). In patients with a tracheostomy, preoperative renal failure was the only perioperative comorbidity found to be significantly associated with DSWI (p = 0.03). Overall operative mortality was 3.6%. Mortality was 16.5% for patients with DSWI, 22.7% for patients requiring tracheostomy, and 55% for patients with DSWI and tracheostomy. CONCLUSIONS: The incidence of DSWI after tracheostomy, in our experience, is not as high as previously reported. Tracheostomy can be performed safely after median sternotomy with a low morbidity and mortality rate. However, if DSWI occurs after tracheostomy, mortality is significant. PMID- 16039217 TI - Numerical simulation techniques to study the structural response of the human chest following median sternotomy. AB - BACKGROUND: The optimal closure technique of median sternotomy remains controversial. The objective of this study was to analyze the structural response of the separated sternum using computer-based numerical discretization techniques, such as finite element methods. METHODS: Thoracic computer tomographic scans (2.5-mm slices) were segmented, analyzed by image processing techniques, and transferred into a three-dimensional finite element model. In a first approach a linear elastic material model was used; neglecting nonlinear and damage effects of the bones. The influence of muscles and tendons was disregarded. Nonlinear contact conditions were applied between the two sternal parts and between fixation wires and sternum. The structural response of this model was investigated under normal breathing and asymmetric leaning on one side of the chest. Displacement and stress response of the segmented sternum were compared regarding two different closure techniques (single loop, figure-of eight). RESULTS: The obtained results revealed that for the normal breathing load case the single loop technique is capable of clamping the sternum sufficiently, assuming that the wires are prestressed. For asymmetric loading conditions, such as leaning on one side of the chest, the figure-of-eight loop can substantially reduce the relative longitudinal displacement between the two parts compared with the single loop. CONCLUSIONS: The application of numerical simulation techniques using complex computer models enabled the determination of structural behavior of the chest regarding the influence of different closure techniques. They allowed easy and fast modifications and therefore, in contrast to a real physical model, in-depth parameter studies. PMID- 16039218 TI - Short operation time: an important element to reduce operative invasiveness in pediatric cardiac surgery. AB - BACKGROUND: The mini skin incision procedure is considered an important element of minimally invasive cardiac surgery because of its definitive cosmetic advantage. However, the operative hazard of limited exposure may be associated with prolonged operation time and increased surgical insult. METHODS: A total of 357 consecutive patients undergoing repair of an isolated atrial or ventricular septal defect, in whom the mini skin procedure was applied, were investigated. Patients were grouped by diagnosis and body weight. Univariate and multivariate risk analyses were conducted in the specific patient group undergoing ventricular septal defect repair weighing less than 5 kg. RESULTS: The operation time was reduced by 21.0% (93.4 to 73.8 minutes) during this time period. Univariate risk analysis revealed that the operation time had a significant correlation with time to extubation (p < 0.0001), catecholamine duration (p = 0.0003), intensive care unit stay (p < 0.0001), hospital stay (p = 0.016), arterio-alveolar oxygen tension difference at the time of extubation (p = 0.0253), and furosemide dose required in the first 24 hours (p = 0.0332). Multiple linear regression analysis revealed that the operation time had an impact on time to extubation, arterio alveolar oxygen tension difference at the time of extubation, and intensive care unit stay. The length of skin incision was not correlated with any outcome measure. CONCLUSIONS: The mini skin incision, if associated with prolonged operation time, may increase the overall insult in pediatric cardiac surgery. In order to reduce operative invasiveness, simultaneous effort to reduce, or at least not to increase, the operation time are mandatory. PMID- 16039219 TI - Arterial switch operation for transposition of the great arteries with coronary arteries from a single aortic sinus. AB - BACKGROUND: The reimplantation of the coronary arteries from a single aortic sinus (single sinus coronary artery) in an arterial switch operation remains a technically challenging procedure. The technique of coronary transfer in this situation should be individualized depending on coronary ostial anatomy. We reviewed our techniques of coronary reimplantation with early and midterm results. METHODS: Among 103 patients who underwent arterial switch operations from March 1994 to June 2004, 16 (15.5%) had single sinus coronary artery (median age, 9 days; mean body weight, 3.5 kg). Fourteen patients (14/16, 87.5%) had coronary arteries from right facing sinus (sinus 2). Of these 14 patients, 11 had separate ostia including intramural course of the left coronary artery (n = 9) and 3 had single ostium. Two patients (2/16, 12.5%) had coronary arteries from left facing sinus (sinus 1) with single ostium. Aortic arch obstruction was associated in 5 patients. RESULTS: All 5 single sinus coronary arteries with single ostium were reimplanted with the trap-door technique. Of the 11 patients with separate ostia, 8 underwent coronary transfer with the aortocoronary flap technique and 3 with the double-button technique. Two of the 3 patients who underwent the double-button technique required left coronary artery bypass using left subclavian artery free graft as the salvaging procedure. There was one early death (1/16, 6.3%), which occurred during our earlier experience, in a patient who had arch anomaly and intramural left coronary artery. There was no late death. All but one patient had good ventricular function. CONCLUSIONS: All single sinus coronary artery with single ostium can be transferred with the trap-door technique with excellent results. The aortocoronary flap technique in those with separate ostia with or without intramural left coronary artery may be a good option. However, reimplantation of the intramural left coronary artery using separate coronary buttons should be performed with great care. PMID- 16039220 TI - Redo mitral valve replacement in children. AB - BACKGROUND: Despite excellent results with current techniques of mitral valve repair, some children still require mitral valve replacement (MVR). Of necessity, a certain percentage of these children need valve rereplacement (redo MVR). METHODS: Of 82 MVRs at our institution since 1988, 22 were redo MVRs in 21 children aged 1.4-21 years (mean 9.8 +/- 5.6 years). Interval from initial MVR was 1 month to 18 years (mean 5.8 +/- 4.9 years). Reoperation indication was primarily stenosis in 13 (usually pannus formation), regurgitation in 4, valve thrombosis or embolism in 4, and endocarditis in 1. The initial valve was mechanical in 18 and xenograft in 4. RESULTS: During redo MVR, 8 of 22 (36%) patients had additional procedures including left ventricular outflow tract obstruction relief or aortic valve replacement in 4 and tricuspid valve repair in 2. The new mitral valve was mechanical in 20 and xenograft in 2. Median valve size increased from 21 mm to 25 mm although 7 valves were not upsized on rereplacement (average valve size increase was 3.3 +/- 1 mm). Only 2 patients (9%) needed a new pacemaker. There were no hospital deaths. Six children developed end-stage heart failure and underwent successful cardiac transplantation 3 to 32 (mean 12.1 +/- 10.0) months, postoperatively. On follow up of 1 month to 7.7 years (mean 2.5 +/- 2.3 years) there has been only one late death of an unrecognized coronary artery anomaly. CONCLUSIONS: Redo MVR in children can be performed safely with low morbidity and mortality. A larger prosthesis can often be placed in these children. A surprisingly high percentage of patients eventually require transplantation, perhaps reflecting unnecessary delay in referral for redo MVR. PMID- 16039221 TI - Results of aortic valve-sparing and restoration with autologous pericardial leaflet extensions in congenital heart disease. AB - BACKGROUND: The purpose of this study is to evaluate the efficacy of aortic valve sparing repair with glutaraldehyde-treated autologous pericardium in congenital valvular pathology. METHODS: Sixty-two patients underwent reparative aortic valve surgery from January 1997 through December 2003. The mean age was 25 +/- 20 years (+/- standard deviation) (range, 10 days to 81 years). Fifty percent (31 of 62) were less than 19 years old at operation. The diagnoses included bicuspid aortic valve (39 patients), ventricular septal defect (14 patients), severe aortic stenosis (6 patients), subaortic stenosis (7 patients), bacterial endocarditis (7 patients), neonatal truncus arteriosus (2 patients), Shone's complex (2 patients), transposition complex (1 patient), double-chambered right ventricle (1 patient), and Marfan's syndrome (1 patient). Twelve patients (19 %) had prior sternotomy and cardiac operations. Valve-sparing techniques included pericardial leaflet extensions in 62 patients, creation of one or more pericardial neoaortic sinuses in 8, subcommissuroplasty in 8, pericardial patch of perforated leaflets in 9, Dacron mesh wrap (Boston Scientific, Wayne, NJ) of dilated ascending aorta in 12, and concomitant tricuspid and mitral valve repairs in 3 and 4 patients, respectively. RESULTS: There was one early death (1.6%). There were no late deaths at a mean follow-up of 25 +/- 16 (range, 0.1 to 72.5 months). Six patients required reoperation and prosthetic or homograft replacement for aortic valve incompetence. One out of 6 reoperations required re-repair. The remaining patients are well with a mean aortic regurgitation grade by echocardiography of 1.3 +/- 0.9 (scale, 0 to 4). CONCLUSIONS: Aortic valve repair with pericardial leaflet extension is a promising technique for the growing child. PMID- 16039222 TI - Right ventricular outflow tract reconstruction with an allograft conduit in non ross patients: risk factors for allograft dysfunction and failure. AB - BACKGROUND: Allograft conduits (AC) are used for reconstruction of the right ventricular outflow tract (RVOT) in children with RVOT hypoplasia or atresia and for pulmonary valve replacement in children undergoing Ross aortic valve replacement (AVR). Children undergoing the Ross AVR are generally older and have their AC inserted in the orthotopic position as opposed to the heterotopic position used for most cases of complex RVOT obstruction. The orthotopic position of the AC combined with the fact that the AC in the Ross patients are larger and can be oversized are the three factors that increase the right ventricular pulmonary artery (RV-PA) conduit durability in the Ross AVR group. A retrospective evaluation of our experience with use of AC in non-Ross patients for reconstruction of the RVOT was conducted to determine the risk factors for failure. METHODS: Between January 1985 and December 2003, 117 non-Ross patients received AC (94 pulmonary and 23 aortic) for RVOT reconstruction. Median age at implantation was 8 months (mean 4.3 +/- 7.1 years; range, 6 days to 43 years). There were 57 children (49%) less than 12 months of age. Endpoints were AC failure (valve explant, balloon dilatation), and AC dysfunction (AC stenosis >40 mm Hg and AC insufficiency more than 2+). There were no device-related deaths. RESULTS: Overall patient survival was 80% at 15 years. Freedom from AC failure was 60% at 5 years and 43% at 15 years. Freedom from failure was worse in infants (42% and 34% at 5 and 15 years, respectively). Freedom from AC dysfunction was 40% at 5 years and 23% at 15 years. Freedom from dysfunction was worse in infants (21% and 16% at 5 and 15 years, respectively). Univariate analysis identified younger patient age, smaller AC size, diagnosis of truncus arteriosus, and the presence of aortic AC as risk factors for AC dysfunction and failure. Multivariate analysis identified smaller AC size and the presence truncus arteriosus as risk factors for AC dysfunction and failure. CONCLUSIONS: Right ventricular outflow tract reconstruction with an AC in non-Ross patients has poor performance at midterm follow-up with AC dysfunction and failure of, respectively, 60% and 40% for the entire group and 79% and 58% in the infant group at 5 years. An alternate conduit for this application must be considered. PMID- 16039223 TI - Specific issues after extracardiac fontan operation: ventricular function, growth potential, arrhythmia, and thromboembolism. AB - BACKGROUND: The purpose of this study was to define the prevalence of specific sequelae after extracardiac Fontan operation. METHODS: Sixty-five consecutive patients undergoing extracardiac Fontan operation were studied for mortality, Fontan failure, systemic ventricular function, supraventricular arrhythmias, thromboembolism, and growth potential. Age was 3 to 31 years (mean +/- standard deviation, 9.4 +/- 1.8; median, 7 years). The conduits were constructed of polytetrafluoroethylene (n = 50), and "viable" in situ pericardium (n = 15). The patients underwent serial echocardiogram, dynamic radionuclide studies, and cardiac catheterization. RESULTS: Operative mortality was 3%, and the incidence of conduit thrombosis was 4.6%. There was paradoxic filling of the right lung after femoral injection of the radiotracer in all cases of conduit obstruction. Perioperative and late postoperative supraventricular arrhythmias were observed in 9.2% and 4.7% of patients, respectively. Risk factors for supraventricular arrhythmias included systemic ventricular dysfunction (p = 0.000), heterotaxy syndrome (p = 0.008), systemic venous anomalies (p = 0.015), and previous bidirectional Glenn operation (p = 0.017). At a mean follow-up of 77 +/- 2 months (range, 8 to 79 months), there were no late deaths (actuarial survival at 79 months, 96.9% +/- 0.02%). Serial echocardiograms demonstrated evidence of growth of the viable tunnels. Postoperatively, there was transient depression of ejection fraction in all patients (p = 0.000). CONCLUSIONS: Supraventricular arrhythmias after extracardiac Fontan are more common in patients with heterotaxy syndrome, bilateral superior venae cavae, systemic ventricular dysfunction, and those undergoing completion Fontan operation. The viable tunnel may emerge as an optimal alternative by virtue of reduction of supraventricular arrhythmias, elimination of the need for anticoagulation, and addressing the issue of growth potential in selected patients. PMID- 16039224 TI - Invited commentary. PMID- 16039225 TI - Minimally invasive midaxillary muscle sparing thoracotomy for atrial septal defect closure in prepubescent patients. AB - BACKGROUND: Partial sternotomy, as well as posterolateral or anterolateral right sided thoracotomy, are used for correction of selected cardiac lesions in children. However, in female patients impaired breast development after an anterolateral thoracotomy is reported, and for both the posterolateral and the anterolateral approach, partial transection of large muscle groups is required. The midaxillary approach may help to avoid these side effects and improve the cosmetic result. METHODS: Beginning in April 2003, our institutional policy changed toward a midaxillary approach in prepubescent patients with an atrial septal defect, in whom criteria for catheter closure were not fulfilled. Thoracotomy was performed after a horizontal midaxillary incision and mobilization of the latissimus dorsi and splitting of the serratus anterior. Aorta and caval veins were cannulated directly. The atrial septal defect was closed during electrically induced fibrillation of the heart. RESULTS: Until August 2004, this technique was applied in 36 patients (30 girls, 6 boys), with no need for conversions to another approach. Mean patient age was 6.9 +/- 2.6 years (range, 4 to 14 years), with a mean weight of 23.8 +/- 11.2 kg (range, 15 to 69 kg). Skin incision ranged from 4.5 to 6.0 cm. Mean cardiopulmonary bypass time was 31 +/- 13 minutes (range, 13 to 73 minutes), with a mean ventricular fibrillation time of 21.2 +/- 7.4 minutes (range, 10 to 42 minutes). In 28 of 36 patients a patch was used. No phrenic nerve damage occurred. CONCLUSIONS: The midaxillary approach is a safe alternative to lateral thoracotomies frequently used in cardiac surgery for atrial septal defect closure. It helps to improve the cosmetic result in the prepubescent patient group. We believe that its application should not be expanded to include repair of more complex lesions or to patients below the age of 3 to 4 years. For these, variations of cosmetically favorable partial sternotomy techniques should be applied. PMID- 16039226 TI - Invited commentary. PMID- 16039227 TI - "Early" delayed sternal closure following pediatric cardiac surgery. AB - BACKGROUND: Delayed sternal closure is commonly used following pediatric cardiopulmonary bypass surgery for many reasons including support of the failing myocardium. We hypothesized that, as a result of improvements in perioperative care, sternal closure could be achieved at an earlier postoperative time than the 3 to 5 days typically reported in the literature. METHODS: Retrospective chart review of all bypass surgery (n = 585) performed in a single center over a 3-year period (2000-2002). RESULTS: We identified 66 children (11.3%), median age 5 days old, who underwent delayed sternal closure. In 60 of these patients, sternal closure was achieved at a median (interquartile) postoperative time of 21 hours (18 to 40 hours). The most common indication was inadequate hemostasis, although early sternal closure was also achieved in the subgroup with poor myocardial function as the primary indication at a median of 36 hours (21 to 44 hours). There was no noticeable hemodynamic, respiratory or metabolic compromise following sternal closure, although patients with poor myocardial function tended to have a lower mean blood pressure than those with inadequate hemostasis (ANOVA, p = 0.02). The overall mortality was 19.7% (13 of 66), with a median duration of ventilation and intensive care stay among survivors of 3.8 days (2.4 to 6.3 days) and 4.8 days (3.7 to 7.9 days), respectively. CONCLUSIONS: Delayed sternal closure is possible at an earlier stage than previously reported. PMID- 16039228 TI - Invited commentary. PMID- 16039229 TI - Single-ventricle physiology reduces cerebral oxygen delivery in a piglet model. AB - BACKGROUND: In single-ventricle physiology, cerebral blood flow and oxygen (O2) delivery may be inadequate. This study tests the hypotheses that in acute univentricular physiology (1) cerebral blood flow increases inadequately to maintain O2 delivery, (2) the brain is incapable of increasing O2 extraction due to hypoxemia, and (3) cerebral O2 delivery diminishes selectively in different brain regions. MATERIAL AND METHODS: Univentricular physiology was created in 8 piglets, while 8 animals were sham controls. Aortopulmonary shunt, echocardiography-guided atrial septostomy, tricuspid valve avulsion, and pulmonary artery occlusion were performed to allow the left ventricle to support systemic and pulmonary circulations. Cerebral blood flow (microspheres), cerebral O2 and lactate metabolism, and cerebral O2 saturation were measured at baseline, 30 minutes, and 120 minutes after conversion to univentricular physiology. RESULTS: Cerebral blood flow increased in the cerebrum and subtentorium in controls (p < 0.05), whereas it remained unchanged in univentricular piglets. Cerebral O2 delivery at 30 and 120 minutes was lower in univentricular physiology than in controls (p = 0.05). Fractional oxygen extraction was unchanged in both groups. Cerebral O2 consumption trended lower in univentricular physiology (p = not significant), while it was unchanged in controls. Lactate cerebral metabolic rate (CMRLactate) increased at 30 and 120 minutes in both groups. The decline in O2 delivery was variable, but present in nearly all brain regions. CONCLUSIONS: This study confirms the hypothesis that, in univentricular physiology, hypoxemia and limited cerebral blood flow reduce cerebral O2 availability in nearly all regions. These findings contribute to the understanding of brain abnormalities in infants with univentricular physiology. PMID- 16039230 TI - The second-generation aortic connector: six months' angiographic follow-up. AB - PURPOSE: Recently, new mechanical anastomotic devices have been developed. Initial results appear to be equivalent to those obtained with suture. The aim of the study was to evaluate the 6-month angiographic patency and clinical results with the St. Jude Medical second-generation aortic connector for proximal aortosaphenous graft anastomosis. DESCRIPTION: From September 2002 to June 2003, 45 connectors were implanted in 39 patients. Thirty-three patients with 36 connectors underwent 6-month angiographic and clinical follow-up. EVALUATION: One connector had an early occlusion and 2 connectors and 1 vein graft were occluded at 6-month angiography, for a patency rate of 88.9% (32 of 36). No device-related complications were detected at 6-month follow-up. CONCLUSIONS: The second generation aortic connector is safe and easy to use. Preliminary results show no device-related complications and a satisfactory 6-month angiographic patency. PMID- 16039231 TI - Invited commentary. PMID- 16039232 TI - A safe and cost-effective approach to minimally invasive radial artery harvesting. AB - PURPOSE: We describe a minimally invasive, cost effective, and safe method of radial artery harvesting. DESCRIPTION: After obtaining informed consent and completing a preoperative evaluation, 169 radial arteries were harvested. Harvesting was accomplished through a 3-cm proximal mid forearm incision with exposure provided by a modified self-retaining lighted retractor. EVALUATION: A total of 169 radial arteries were successfully harvested. The average incision length was 2.9 cm, radial artery length was 15.8 cm, and harvest time was 32.7 minutes. No trauma to the artery or graft spasm was evident. No procedure required conversion to an open technique. Superficial cellulitis occurred in 2 patients (1.2%) and wound infection in 1 (0.6%). Three patients (1.8%) experienced intermittent residual dysesthesia. All of the patients were highly satisfied with the excellent aesthetic results. This approach allowed for a substantial cost savings compared with other minimally invasive techniques. CONCLUSIONS: Direct minimally invasive radial artery harvesting is an acceptable alternative approach to radial artery harvesting. This method is safe, cost effective, easily reproducible, and aesthetically pleasing. PMID- 16039233 TI - A stentless trileaflet valve from a sheet of decellularized porcine small intestinal submucosa. AB - PURPOSE: The purpose of this study was to investigate the function of a trileaflet pulmonary valve constructed from a sheet of porcine small intestinal submucosa. DESCRIPTION: In four sheep, the native pulmonary valve and a segment of the pulmonary trunk was excised and replaced with a trileaflet valve constructed from decellularized porcine small intestinal submucosa. The valve construct was created from a sheet of the xenograft material by a method of involuting flaps of tissue inside a cylinder of itself. The function of the valve was assessed by echocardiography, catheter pullback across the valve, and observation of an excised valve in a flow simulator. EVALUATION: The valve constructs exhibited low gradients and symmetrical leaflet movement with good mobility when tested under physiologic conditions in an acute sheep model. CONCLUSIONS: This method offers a means to create a functional trileaflet valve replacement from a sheet of tissue. PMID- 16039234 TI - Invited commentary. PMID- 16039235 TI - Virtual vascular endoscopy for acute aortic dissection. AB - Virtual endoscopy of the aorta is a new three-dimensional reconstruction method from multislice computed tomography or magnetic resonance that offers a virtual navigation through the aorta, and the possibility of having a new preoperative endoluminal vision. We present a case of subacute aortic dissection with a preoperative virtual endoscopy of the aorta. PMID- 16039236 TI - Traumatic rupture of a descending thoracic aortic homograft. AB - My colleagues and I report an unusual case of traumatic aortic injury in an 18 year-old woman who had undergone multiple prior surgical procedures for repair of a type B interrupted aortic arch. Her most recent procedure included replacement of the proximal descending thoracic aorta with a 19-mm homograft at age 11 years. Seven years later, she was involved in a motor vehicle collision after a syncopal episode. Imaging studies revealed rupture of the body of the aortic homograft with formation of a pseudoaneurysm. The injury was successfully repaired with a Dacron graft by using hypothermic circulatory arrest. PMID- 16039237 TI - Cell therapy plus transmyocardial laser revascularization for refractory angina. AB - We describe the use of autologous bone marrow cells combined with transmyocardial laser revascularization in a 74-year-old man with refractory angina. Baseline cardiac magnetic resonance imaging revealed a markedly depressed left ventricle systolic function and an extensive area of myocardial ischemia. During surgery, 11 laser shots using a CO2 Heart Laser System (PLC Medical Systems, Milford, MA) were fired and a 5-mL cell suspension containing 21.5 x 10(6) bone marrow cells/mL was delivered by multiple injections into the myocardium. At 6 months after the procedure, another cardiac magnetic resonance imaging showed an almost complete resolution of the perfusion defect and an improvement in left ventricular contractility. PMID- 16039238 TI - Severe hypothyroidism after coronary artery bypass grafting. AB - A 56-year-old man with unstable angina underwent urgent coronary artery bypass grafting. The patient required reintubation and prolonged ventilation because of persistent drowsiness and hypotension. The patient was weaned off the ventilator and extubated; however, he remained drowsy and lethargic. Neurologic examination, electroencephalogram, and computed tomography scan of the brain did not show any organic cause of his depressed neurologic status. His metabolic profile revealed severe hypothyroidism. The patient responded well to oral thyroxine. We report the unusual manner in which hypothyroidism presented in the patient. A high index of suspicion is required to diagnose and treat this complication. PMID- 16039239 TI - Complete atrioventricular block decreases left ventricular assist device flow rate. AB - A 52-year-old man, living at home, had been on left ventricular assistance for 15 months with a Novacor pump (Worldheart, Ottawa, ON, Canada). Following the onset of effort dyspnea that had recently become worse, he was admitted to the hospital. A review of the pump showed that its flow rate had significantly decreased. An electrocardiogram showed a third-degree atrioventricular block. A pacemaker was implanted and the patient became asymptomatic immediately afterwards. This clinical case report shows the importance of satisfactory right ventricle filling in these patients. Right ventricle filling can be impaired by conduction disorders. PMID- 16039240 TI - Left ventricular assist device as destination therapy in doxorubicin-induced cardiomyopathy. AB - Doxorubicin-induced cardiomyopathy is not uncommon and may progress to end-stage heart failure. Treatment of this condition with heart transplantation, however, requires that the primary malignancy be deemed "cured." We present the case of a 55- year-old woman who had doxorubicin-induced cardiomyopathy and non-Hodgkin's lymphoma. The active status of her lymphoma precluded heart transplantation. She had end-stage heart failure and underwent the insertion of a left ventricular assist device as a destination therapy. PMID- 16039241 TI - Successful management of two heart transplant recipients with mycobacterial pulmonary infections. AB - We report two cases of pulmonary tuberculosis in heart transplant recipients: a 46-year-old man with pulmonary tuberculosis due to Mycobacterium tuberculosis and a 64-year-old man with nontuberculous mycobacterial infection with pulmonary infiltrates due to Mycobacterium xenopi. The time intervals from transplantation to diagnosis were 3 and 4 years, respectively. The patient with tuberculosis underwent standard treatment with isoniazid, rifampin, ethambutol, and pyrazinamide. The patient with the nontuberculous mycobacterial infection received treatment with clarithromycin and ciprofloxacin for 18 months in addition to rifampin for the first 3 months. Both patients responded well to treatment. No recurrences were observed during follow-up. The interactions between antibiotic treatment and cyclosporine therapy should be observed closely in organ transplant recipients, requiring frequent level determinations and dosing changes. PMID- 16039242 TI - Sliding plasty using the triple-orifice technique for tricuspid endocarditis. AB - A 29-year-old male with tricuspid endocarditis related to a residual ventricular septal defect was studied. A large vegetation was resected along with partial excision of the valve. The tricuspid valve was reconstructed using a single stitch triple-orifice technique. No regurgitation or stenosis was detected at the valve 6 months after surgery. PMID- 16039243 TI - A fatal fungus. AB - We report the case of a 66-year-old man who presented with a prodromal type illness and pre-sternal swelling after having coronary artery bypass grafts 4 years earlier. Computed tomography showed that the mass had a retrosternal extension, to join a collection anterior to the right ventricle. Candida famata was isolated from operative specimens and he was treated with aggressive antifungal therapy. We believe that this is the first reported case of mediastinal Candida famata. PMID- 16039244 TI - Rupture of a saphenous vein coronary artery bypass graft due to Aspergillus necrotizing vasculitis. AB - We present the first, unusual case of a lethal mediastinal hemorrhage caused by rupture of a saphenous vein aortic coronary bypass graft due to Aspergillus species necrotizing vasculitis in an immunocompetent patient 18 days after redo coronary artery bypass surgery. The patient had neither signs for mediastinitis nor for another source of Aspergillus infection. PMID- 16039245 TI - Mycotic aneurysm of the descending thoracic aorta in a 2-kg neonate. AB - Umbilical artery catheterization is often performed in critically ill neonates. Infection and thromboembolic events are relatively frequent complications, but aneurysm formation is rare. We describe the case of a premature baby who developed a rapidly expanding aneurysm of the descending thoracic aorta, secondary to a highly placed infected umbilical catheter. The rapid progression under medical therapy prompted us to replace the thoracic aorta with an 8-mm polytetrafluoroethylene graft, using extracorporeal circulation and core cooling. The baby had an excellent recovery and was discharged home after a prolonged antibiotic course. He is being followed with repeated imaging studies for a small abdominal aneurysm. We describe a novel approach to this rare and complicated problem and provide a review of the literature on the subject. PMID- 16039246 TI - Coarctation of the thoraco-abdominal aorta associated with mucopolysaccharidosis VII in a child. AB - Herein we describe a case of atypical coarctation of the thoraco-abdominal aorta associated with mucopolysaccharidosis VII in a 4-year-old girl. Aortography showed diffuse narrowing of the descending aorta. An extra-anatomic bypass was constructed using an 8-mm Dacron graft (Meadox Medicals Inc, Oakland, NJ) between the proximal and distal portion of the descending aorta. Balloon angioplasty was necessary to treat stenoses in the infrarenal abdominal aorta. Two years postoperatively at age 6, aortography revealed no stenosis at the anastomotic sites or in the prosthesis, but the hypoplastic segment of the descending aorta between the anstomoses was completely occluded. PMID- 16039247 TI - Aortobronchial fistula in a pediatric patient with massive hemoptysis: treatment by means of an aortic endograft. AB - We present an 11-year-old girl with acute myelogenous leukemia and hemoptysis from abscess erosion into the descending thoracic aorta. We report a pediatric case of an aortobronchial fistula treated with an aortic endograft and discuss the technical limitations and potential complications of this procedure. PMID- 16039248 TI - Complete correction of interrupted right aortic arch with isolation of left subclavian artery. AB - Isolation of the left subclavian artery is a rare aortic arch anomaly. Association with a right interrupted aortic arch and a double patent ductus arteriosus is exceptional. We report a case of such an association with successful surgical correction. PMID- 16039249 TI - Simultaneous second-stage norwood operation and tracheoplasty. AB - Tracheal stenosis is a rare complication that follows operations for congenital heart disease in the newborn period. We report the case of an infant with hypoplastic left heart syndrome who developed symptomatic tracheal stenosis 5 months after a first-stage Norwood operation. The combined surgical procedure of a bidirectional Glenn anastomosis and a tracheoplasty under cardiopulmonary bypass was undertaken. PMID- 16039250 TI - Surgical treatment of intramyocardial foregut cysts. AB - Intramyocardial foregut cysts have been identified as incidental findings at autopsies, but there have been no reports of surgical management of clinically significant cysts. We report 2 patients and provide recommendations for appropriate management. PMID- 16039251 TI - Stiff man syndrome with thymoma. AB - Paraneoplastic stiff man syndrome with a thymoma is rare disease. We treated a 57 year-old woman with a type B1 thymoma, based on the World Health Organization classification, who had stiff man syndrome. Her symptoms were alleviated after a thymectomy. Herein we report a case of stiff man syndrome with a thymoma and also review three cases reported previously. PMID- 16039252 TI - Papillary carcinoma of the thymus gland. AB - In 1999, the World Health Organization introduced a new classification of a rare thymic cancer called papillary adenocarcinoma of the thymus gland. In this study we report on a case of papillary thymic carcinoma in a 29-year-old woman. Histologically, the tumor consisted of a papillary component admixed with a solid component, a component that is commonly found in type A thymoma. PMID- 16039253 TI - Magnetic resonance imaging evaluation of surgical ventricular reconstruction. PMID- 16039254 TI - Saccular aneurysm at the aortic isthmus. PMID- 16039255 TI - Anterior approach to implant the Jarvik 2000 with retroauricular power supply. AB - The retroauricular power supply of the Jarvik 2000 (Jarvik Heart Inc, New York, NY) left ventricular assist device is suitable for permanent support, as it is associated with fewer infections than conventional drivelines. Implantation through a left-lateral thoracotomy limits the performance of additional cardiac procedures. We describe a technique that used a sternotomy for the implantation of the Jarvik 2000 with retroauricular power supply in two patients. The retroauricular power supply of the Jarvik 2000 can be provided with an anterior approach, allowing full surgical access to the heart. If the outflow graft to the ascending aorta indeed reduces aortic stasis and thromboembolic events, the anterior approach with retroauricular power delivery might evolve into a standard procedure. PMID- 16039256 TI - The Florida sleeve: a new technique for aortic root remodeling with preservation of the aortic valve and sinuses. AB - Aortic valve salvage remains the procedure of choice for patients with incompetent valves and normal leaflet morphology. Here my colleagues and I describe a new technique for aortic root remodeling with preservation of the aortic valve. PMID- 16039257 TI - Minimally invasive surgical alternatives for left ventricle epicardial lead implantation in heart failure patients. AB - Biventricular pacing is a proven advantageous adjuvant therapy for patients with ventricular dyssynchrony associated with congestive heart failure. Endocardial left ventricular lead placement does have several limitations: anatomic variations of the coronary venous system and late lead dislodgement. Epicardial lead placement is often a rescue procedure but offers some advantages related to safety and a shorter implant time. Moreover, it allows visual selection of the best pacing site and multiplicity of pacing sites. Three minimally invasive surgical methods of left ventricular lead placement are outlined in this article, including specific indications and limitations. Biventricular pacing has been proposed as an adjuvant treatment for patients with heart failure and intraventricular conduction delay. PMID- 16039258 TI - Thoracoscopic rib resection using a Gigli saw. AB - When a conventional approach is used to perform a rib resection, a skin incision longer than the rib to be resected must be made. As a result, a conventional rib resection leaves a rather large and esthetically unfavorable scar. After considering pain management, esthetics, and quality of life, we devised a new technique for thoracoscopic rib resection that uses a Gigli saw. This new technique was performed on an overweight woman with a solitary metastatic bone tumor of the right eighth rib, whose case is described herein. The patient's postoperative course was satisfactory. Since this technique does not require a long skin incision, pain management and aesthetic results are improved compared with conventional techniques. PMID- 16039259 TI - Effect of cardioplegic and organ preservation solutions and their components on coronary endothelium-derived relaxing factors. AB - Cardioplegic (and organ preservation) solutions were initially designed to protect the myocardium (cardiac myocytes) during cardiac operation (and heart transplantation). Because of differences between cardiac myocytes and vascular (endothelial and smooth muscle) cells in structure and function, the solutions may have an adverse effect on coronary vascular cells. However, such effect is often complicated by many other factors such as ischemia-reperfusion injury, temperature, and perfusion pressure or duration. To evaluate the effect of a solution on the coronary endothelial function, a number of points should be taken into consideration. First, the overall effect on endothelium should be identified. Second, the effect of the solution on the individual endothelium derived relaxing factors (nitric oxide, prostacyclin, and endothelium-derived hyperpolarizing factor) must be distinguished. Third, the effect of each major component of the solution should be investigated. Lastly, the effect of a variety of new additives in the solution may be studied. Based on available literature these issues are reviewed to provide information for further development of cardioplegic or organ preservation solutions. PMID- 16039260 TI - Balancing the benefit and risk of oral antiplatelet agents in coronary artery bypass surgery. AB - Concern about possible hemorrhagic complications arising from use of oral antiplatelet agents in immediate proximity to coronary artery bypass graft (CABG) surgery leads many clinicians to avoid or discontinue these agents preoperatively. Recent evidence suggests that aspirin and clopidogrel can be used with relative safety in the preoperative period; dual antiplatelet therapy in the 5 days immediately preceding CABG surgery results in a moderate and variable increase in the risk of procedural bleeding. This modest hemorrhagic risk may be acceptable, given the clinical benefits of sustained antiplatelet therapy in preventing graft occlusion and ischemic complications pre- and post-CABG. Because the bleeding risk with aspirin is dose dependent, use of a low dose is preferred post-CABG. PMID- 16039261 TI - Screening of peripheral vascular disease in patients facing coronary surgery: is invasive angiography really out? PMID- 16039262 TI - Mechanical bioprosthesis. PMID- 16039264 TI - Menopausal status and non-small cell lung cancer. PMID- 16039265 TI - Orthotopic cardiac transplantation: modification of bicaval anastomosis. PMID- 16039266 TI - Is identification of prognostic factors for lung metastasectomy sufficient to establish selection criteria? PMID- 16039269 TI - Neuroapraxia. PMID- 16039271 TI - Substitution of a conserved alanine in the domain IIIS4-S5 linker of the cardiac sodium channel causes long QT syndrome. AB - OBJECTIVE: Congenital long QT syndrome type 3 (LQT3) is an inherited cardiac arrhythmia disorder due to mutations in the cardiac sodium channel gene, SCN5A. Although most LQT3 mutations cause a persistent sodium current, increasing diversity in the disease mechanism is shown. Here we present the electrophysiological properties of the A1330T sodium channel mutation (DIIIS4-S5 linker). Like the A1330P, LQT3 mutation, A1330T, causes LQT3 in the absence of a persistent current. METHODS: A1330T, A1330P and wild-type sodium channels were expressed in HEK-293 cells and characterized using the whole-cell configuration of the patch-clamp technique. RESULTS: The A1330T mutation shifts positively the voltage-dependence of inactivation and speeds recovery from inactivation. Measurements of sodium window (I(Na, window)) currents revealed a positive shift of the I(Na, window) voltage range for both 1330 mutants, with in addition an increase in I(Na, window) magnitude for the A1330P mutant. Action potential (AP) clamp experiments revealed that these changes in I(Na, window) properties cause an increased inward current during the initial part of phase 4 repolarization of the AP. CONCLUSIONS: Our findings indicate that the alanine at position 1330 in the DIIIS4-S5 linker of the cardiac sodium channel has a role in channel fast inactivation. Substitution by a threonine shifts the voltage range of I(Na, window) activity to more positive potentials. Here the counter-acting effect of outward K+ current is reduced and may delay AP repolarization, explaining the LQT3 phenotype. PMID- 16039272 TI - Modulation of I(Kr) inactivation by mutation N588K in KCNH2: a link to arrhythmogenesis in short QT syndrome. AB - OBJECTIVE: Short QT syndrome (SQTS) is characterized by ventricular arrhythmias and sudden death. One form of SQTS is caused by mutation N588K in human ether-a go-go-related gene (HERG). In this study we sought to determine the potential role of N588K in arrhythmias. METHODS: We measured the characteristics of HERG current generated by wild-type (WT) KCNH2 and the N588K mutant channel expressed in mammalian TSA201 cells. RESULTS: Whole-cell patch-clamp recordings of WT HERG currents showed the usual rapid onset of inactivation (rectification) at potentials more positive than +10 mV. In contrast, N588K currents rectified at potentials over +80 mV. Over the physiological range of potentials, N588K currents do not inactivate. During an action potential clamp, WT currents displayed a "hump" like waveform with slow activation kinetics and a rapid increase during phase 3 repolarization. In contrast, N588K currents were proportional to the amplitude of the action potential and displayed a dome-like configuration and a much larger current during the initial phases in the ventricle. Purkinje cell action potentials display a more negative phase 2 repolarization than the ventricle and elicited much smaller WT and N588K currents of similar amplitudes. CONCLUSIONS: Physiologically the N588K mutation abolishes rectification of HERG currents and specifically increases I(Kr) in the ventricle with minimal effects on the Purkinje fiber action potential duration. Such preferential prolongation may explain the separation of the T and U waves observed in the ECG of SQTS patients and lead to re-excitation of the ventricle endocardium. PMID- 16039273 TI - Atrial fibrillation-associated minK38G/S polymorphism modulates delayed rectifier current and membrane localization. AB - BACKGROUND: Atrial fibrillation (AF) is a common acquired arrhythmia with multi factorial pathogenesis. Recently, a single nucleotide polymorphism (SNP, A/G) at position 112 in the KCNE1 gene, resulting in a glycine/serine amino acid substitution at position 38 of the minK peptide, was associated with AF occurrence (AF more frequent with minK38G); however, the functional effect of this SNP is unknown. METHODS AND RESULTS: We used patch clamp recording, confocal microscopy and protein biochemistry to study the effect of this SNP on delayed rectifier current expression and mathematical simulation to identify potential functional consequences. The density of slow delayed rectifier current (I(Ks)) resulting from co-expression with KvLQT1 was smaller with minK38G (e.g. at +10 mV: 50+/-7 pA/pF in Chinese hamster ovary (CHO) cells, 45+/-14 pA/pF for COS-7 cells) compared to minK38S (93+/-17 pA/pF, 104+/-23 pA/pF, respectively, P<0.05 for each). I(Ks) kinetics and voltage-dependence were unaffected. Currents resulting from co-expression of human ether-a-go-go-related gene (HERG) were similar for minK38G and minK38S, e.g. upon repolarization from +10 to -50 mV: tail currents 23+/-4 pA/pF versus 22+/-5 pA/pF (P=ns). KvLQT1 membrane immunofluorescence was less in CHO cells co-expressing minK38G versus minK38S, and surface expression of KvLQT1, as determined by labelling with streptavidin/biotin, was increased with minK38S co-expression. Computer simulations with a human atrial action potential model predicted that the minK38G SNP would slightly prolong the atrial action potential and reduce the frequency for alternans behaviour. In the presence of reduced repolarization reserve, these effects were enhanced and under specific conditions early afterdepolarizations occurred. CONCLUSIONS: The minK38G isoform is associated with reduced I(Ks), likely due to decreased KvLQT1 membrane expression. This study reveals a novel amino acid determinant of the minK-KvLQT1 interaction, and if the role of minK38G in AF is confirmed, would suggest mechanistic heterogeneity in genetic determinants of AF. PMID- 16039274 TI - In vitro molecular interactions and distribution of KCNE family with KCNQ1 in the human heart. AB - OBJECTIVE: The voltage-gated K+ channel KCNQ1 associates with the small KCNE1 beta subunit to underlie the IKs repolarizing current in the heart. Based on sequence homology, the KCNE family is recognized to comprise five members. Controversial data have indicated their participation in several K+ channel protein complexes, including KCNQ1. The expression level and the putative functions of the different KCNE subunits in the human heart still require further investigation. METHODS: We have carried out a comparative study of all KCNE subunits with KCNQ1 using the patch-clamp technique in mammalian cells. Real-time RT-PCR absolute quantification was performed on human atrial and ventricular tissue. RESULTS: While KCNQ1/KCNE1 heteromultimer reached high current density with slow gating kinetics and pronounced voltage dependence, KCNQ1/KCNE2 and KCNQ1/KCNE3 complexes produced instantaneous voltage-independent currents with low and high current density, respectively. Co-expression of KCNE4 or KCNE5 with KCNQ1 induced small currents in the physiological range of voltages, with kinetics similar to those of the KCNQ1/KCNE1 complex. However, co-expression of these inhibitory subunits with a disease-associated mutation (S140G-KCNQ1) led to currents that were almost undistinguishable from the KCNQ1/KCNE1 canonical complex. Absolute cDNA quantification revealed a relatively homogeneous distribution of each transcript, except for KCNE4, inside left atria and endo- and epicardia of left ventricular wall with the following abundance: KCNQ1 >> KCNE4 > or = KCNE1 > KCNE3 > KCNE2 > KCNE5. KCNE4 expression was twice as high in atrium compared to ventricle. CONCLUSIONS: Our data show that KCNQ1 forms a channel complex with 5 KCNE subunits in a specific manner but only interactions with KCNE1, KCNE2, and KCNE3 may have physiological relevance in the human heart. PMID- 16039275 TI - Sialic acid as a protective barrier against neointima development. AB - Arterial sialic acid (SA) has been shown to attenuate the binding of fibrinogen and low-density lipoproteins (LDL) to the vessel wall, presumably protecting against atherosclerosis. This study was aimed to assess the effect of changes in SA content in intimal thickening, an early step in the development of atherosclerosis. New Zealand white rabbits were subjected to bilateral carotid periarterial collaring, followed by in situ-perfusion with neuroaminidase (random artery) and with vehicle (contralateral control artery). The efficiency of SA removal was evaluated in perfusates and arterial homogenates, and arterial tissue samples were obtained 7 and 14 days after the intervention to assess morphological changes. Neuraminidase significantly reduced SA by 16.7%. Arterial desialylation was associated with a significantly increased neointimal formation. Proliferation of smooth muscle cells (SMCs), assessed by incorporation of bromo 2'-deoxyuridine into replicating DNA was also significantly increased in desialylated arteries. In addition, immunohistochemical studies showed a slightly stronger oxidized-LDL (ox-LDL) immunostaining in neointima of desialylated arteries than in control vessels. A mild reduction of SA increases intimal thickening, at least partly due to an enhanced proliferation of SMCs, and may facilitate the accretion of atherogenic lipoproteins, providing evidence for the potential role of SA in the protection against neointimal development. PMID- 16039276 TI - Pioglitazone increases the fractional catabolic and production rates of high density lipoproteins apo AI in the New Zealand White Rabbit. AB - Pioglitazone is an agonist of the peroxisome proliferator-activated receptor gamma (PPARgamma) that raises HDL-cholesterol plasma in humans. Whether pioglitazone-mediated modifications in HDL-apolipoprotein AI (apo AI) turnover in vivo contribute to this effect has not been completely elucidated. Therefore, we performed kinetic studies of HDL-apo AI radiolabeled with 125I in male New Zealand White rabbits after 6 weeks of 0.6 (n = 8), 1.75 (n = 8), and 2.6 mg/kg/day (n = 7) pioglitazone and vehicle (n = 12) treatment. Fractional catabolic rate (FCR) of HDL-apo AI was significantly higher in 1.75 and 2.6 mg/kg pioglitazone-treated animals, as compared with control rabbits (0.057+/-0.014 and 0.049+/-0.01 versus 0.025+/-0.005 pools/h, respectively); these changes were associated to a similar increase in apo AI production rates (PR) (1.24+/-0.62 and 1.14+/-0.40 versus 0.53+/-0.17 mg/kg/h, p < 0.01). Consequently, apo AI plasma levels in pioglitazone-treated animals were similar to those of controls. The apo AI-FRC and -PR correlated with the relative proportion of the HDL3c subclass, as determined by polyacrylamide gradient electrophoresis. Our data demonstrate that pioglitazone markedly modifies apo AI kinetics and enhances the proportion of small HDL3c particles, despite the unchanged apo AI concentration. Whether or not the pioglitazone-induced structural changes of HDL contribute to the anti atherosclerotic effects of the drug remains to be determined. PMID- 16039277 TI - C-Type natriuretic peptide and natriuretic peptide receptors are expressed by smooth muscle cells in the neointima after percutaneous coronary intervention. AB - Understanding restenosis after percutaneous coronary intervention (PCI) remains a challenge. Neointimal proliferation is the main cause of restenosis. C-Type natriuretic peptide (CNP) plays a role in relaxation and growth inhibition of vascular smooth muscle cells (SMCs); the effects depend on the presence of specific natriuretic peptide receptors (NPRs) consisting of NPR-A, NPR-B, and NPR C. To test the hypothesis that CNP and NPRs may be involved in restenosis, we immunohistochemically studied the expression of CNP and NPRs during the post-PCI healing process; 10 sites after PCI obtained at autopsy and 14 atherectomy specimens obtained from restenotic sites were investigated. Frozen sections were stained with antibodies against CNP, NPRs, SMCs, macrophages, and endothelial cells. Within 2 months after PCI, most neointimal SMCs expressed CNP and NPR-A. The expression of CNP and NPR-A in these neointimal SMCs decreased from 6 months onward. In contrast, NPR-C was strongly expressed in neointimal SMCs from 1 to 9 months after PCI. In atherectomy specimens, most neointimal SMCs showed weak positivity for CNP and NPR-A, but NPR-C was strongly expressed in the neointimal SMCs. These findings strongly suggest that a paracrine and autocrine system of CNP and NPRs may be important in controlling neointimal growth after PCI in humans. PMID- 16039278 TI - Increased hypercholesterolemia and atherosclerosis in mice lacking both ApoE and leptin receptor. AB - Diabetes mellitus is one of the major risk factors associated with atherosclerosis and coronary heart disease but the mechanistic links between the disease and atherosclerosis are not well understood. In this study, we investigated the effect of the deletion of the long-form leptin receptor on the progression of atherosclerosis in ApoE-/- mouse. ApoE-/-;db/db double knockout mice as well as ApoE-/-;db/+ and ApoE-/- littermates were generated by crossing ApoE-/- and db/+ mice. On a regular chow diet, ApoE-/-;db/db mice at 20 weeks of age exhibited features typical of type 2 diabetes: obesity, hyperglycemia, hyperinsulinemia and dyslipidemia and had significantly accelerated atherosclerosis compared with their age-matched ApoE-/- littermates as assessed by either the percentage of the aorta bearing lesion (5.3+/-0.9% for ApoE-/ ;db/db versus 1.5+/-0.5% for ApoE-/-) or by aortic lipid content ( approximately 1.5-2-fold increase in free cholesterol and approximately 3-4-fold increase in cholesteryl ester). The atherosclerosis in these ApoE-/-;db/db mice was further accelerated by feeding mice with a Western diet and markedly inhibited by fenofibrate with a 2.5- and 5.3-fold reduction of the lesion in male and female mice, respectively. The results from this study demonstrate that type 2 diabetes can accelerate atherogenesis in mice. This mouse model may provide insight into the mechanistic link between type 2 diabetes and atherosclerosis as well as serve as a valuable tool for evaluating therapeutics. PMID- 16039279 TI - AAV serotype-dependent apolipoprotein A-I Milano gene expression. AB - Recent evidence from a double-blind, randomized study showed that treatment with apolipoprotein A-I Milano (ApoA-I Milano) in a complex with phospholipids produced significant regression of the coronary atheroma burden in patients with acute coronary syndromes. We previously showed similar regression of atherosclerosis in an animal model. Here, we examined a viral vector-based gene delivery system as a basis for ApoA-I Milano gene therapy. Comparing levels of expression using combinations of the cytomegalovirus (CMV) promoter in a recombinant serotype 2 adeno-associated virus (rAAV2) linked to ApoA-I Milano or the enhanced green fluorescent protein (EGFP) genes, we found that a promoter construct of two CMV core promoters sharing a CMV enhancer was more active than other combinations or a single CMV promoter. In vivo assessment of this optimal CMV construct using rAAV2 virus particles for intravenous (IV) or intramuscular (IM) routes of delivery produced high circulating levels of ApoA-I Milano protein for extended periods (up to 220 ng/ml at 22 weeks p.i.) by IV delivery while the IM route resulted in a relatively short period of very low-level ApoA-I Milano expression. Since there was no difference in the immune response between the two routes of delivery, we reasoned that tissue tropism might be responsible for this differential gene expression. To explore this possibility, we investigated the effect of different AAV serotypes on ApoA-I Milano gene expression in vivo. It found that rAAV1-mediated expression of ApoA-I Milano was approximately 15- and 9 fold higher than rAAV2 and rAAV5, respectively when IM injection routes were compared while all three AAV serotypes produced substantial levels of ApoA-I Milano expression from IV injection. These studies demonstrate that by modifying the promoter and serotype, increases in the efficiency of AAV-directed transgene expression could be achieved and support the potential of AAV-mediated gene therapy. PMID- 16039280 TI - Tissue-specific expression pattern of human endothelial lipase in transgenic mice. AB - Endothelial lipase (EL), a new member of the triacylglycerol lipase gene family, is a key enzyme in HDL metabolism. The EL expression pattern in humans was reported to be unique and complementary to that documented for lipoprotein lipase. The regulatory elements responsible for the tissue-specific EL expression are not identified yet. In order to confine these sequences to a defined region of the EL promoter, we analyzed EL mRNA expression in human EL transgenic mice expressing EL under the control of the endogenous human promoter. We identified small intestine, mammary gland, adipose tissue and the adrenal gland as previously unknown tissues to express EL. Our data demonstrate that regulatory elements within 11.4 kb of 5' and 9.9 kb of 3' human EL flanking region promote the expression of EL in small intestine, ovary, testis, mammary gland, brain, lung, aorta, adipose tissue and the adrenals, whereas regulatory sequences located between 27.4 and 11.4 kb of 5' or 9.9 and 48.7 kb of 3' human EL flanking region seem to be responsible for kidney-specific EL expression. PMID- 16039281 TI - The plasminogen activator inhibitor-1 (PAI-1) promoter haplotype is related to PAI-1 plasma concentrations in lean individuals. AB - BACKGROUND: Elevated plasminogen activator inhibitor-1 (PAI-1) concentrations are associated with cardiovascular diseases. PAI-1 antigen levels are influenced by environmental factors such as body mass index (BMI), and by genetic factors. The PAI-1 promoter of the PAI-1 gene contains two common polymorphisms (-844A/G and 675(4G/5G)) and the 4G allele of the -675(4G/5G) variation has been associated with elevated PAI-1 concentrations and on some occasions with an increased risk of cardiovascular disease. OBJECTIVES: The aim of our study was to investigate the effect of the PAI-1 promoter haplotype on PAI-1 concentrations and to determine the role of BMI. METHODS: The association between the PAI-1 promoter haplotype and PAI-1 antigen levels was investigated in two independent populations, each including 600 healthy Caucasians. Furthermore, to assess the effect of the PAI-1 promoter haplotype on PAI-1 promoter activity, in vitro reporter gene assays were performed in HepG2 and BAEC cells. RESULTS: We observed significantly higher PAI-1 concentrations in A-4G homozygotes than in G-5G carriers in lean subjects (BMI in the lowest quartile). In these lean subjects, the PAI-1 concentrations in A-4G/G-5G heterozygotes were reduced to 60-75%, and the concentrations in G-5G homozygotes to 45-55%, compared to the PAI-1 concentrations of A-4G homozygotes (p < 0.01). PAI-1 concentrations increased approximately four-fold from the lowest to the highest BMI quartile (p < 0.01). The reporter gene assays did not support a direct effect of the PAI-1 promoter haplotype on promoter activity in HepG2 or BEAC cells. CONCLUSIONS: Our study suggests that the PAI-1 promoter haplotype and BMI affect PAI-1 concentrations and that BMI is a stronger determinant than PAI-1 promoter variation. PMID- 16039282 TI - Leukocyte subpopulations and arteriogenesis: specific role of monocytes, lymphocytes and granulocytes. AB - OBJECTIVE: Circulating leukocytes play a crucial role during arteriogenesis. However, known pro-arteriogenic compounds (MCP-1, GM-CSF) acting via monocytic pathways also exert positive effects on granulocytes and lymphocytes. The role of these two cell types in arteriogenesis remains yet to be clarified, which was the aim of the current study. METHODS: Ninety New Zealand White Rabbits received either phosphate buffered saline (PBS), monocyte chemoattractant protein-1 (MCP 1), interleukin-8 (IL-8), neutrophil activating protein-2 (NAP-2) or lymphotactin (Ltn) via osmotic minipumps after unilateral femoral artery ligation. In vitro stimulation and in vivo assessment of chemoattraction confirmed cell-specific action of the compounds in rabbits. Arteriogenesis was evaluated by angiography and collateral conductance measurements using fluorescent microspheres. Quantitative immunohistology was used to quantify transmigrated leukocyte subtypes after infusion of the factors. RESULTS: MCP-1 infusion attracts monocytes and granulocytes, whereas IL-8 attracts all three cell types albeit monocytes to a significantly lower degree than MCP-1. NAP-2 and lymphotactin selectively attract granulocytes, respectively, lymphocytes. Of the tested cytokines, only MCP-1 stimulates arteriogenesis, as assessed by collateral conductance measurements ((ml/(min 100 mmHg)): PBS, 50.70+/-5.15; MCP-1, 216.30+/ 12.30; IL-8, 58.91+/-5.56; NAP-2, 66.83+/-8.72; Ltn, 52.80+/-5.37) and angiographic findings. CONCLUSION: This study for the first time provides evidence that not granulocytes or T-lymphocytes but monocytes are the key mediators of arteriogenesis. PMID- 16039283 TI - In vivo detection of macrophages in a rabbit atherosclerotic model by time resolved laser-induced fluorescence spectroscopy. AB - Accumulation of numerous macrophages in the fibrous cap is a key identifying feature of plaque inflammation and vulnerability. This study investigates the use of time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) as a potential tool for detection of macrophage foam cells in the intima of atherosclerotic plaques. Experiments were conducted in vivo on 14 New Zealand rabbits (6 control, 8 hypercholesterolemic) following aortotomy to expose the intimal luminal surface of the aorta. Tissue autofluorescence was induced with a nitrogen pulse laser (337 nm, 1 ns). Lesions were histologically classified by the percent of collagen or macrophage foam cells as well as thickness of the intima. Using parameters derived from the time-resolved fluorescence emission of plaques, we determined that intima rich in macrophage foam cells can be distinguished from intima rich in collagen with high sensitivity (>85%) and specificity (>95%). This study demonstrates, for the first time, that a time-resolved fluorescence-based technique can differentiate and demark macrophage content versus collagen content in vivo. Our results suggest that TR-LIFS technique can be used in clinical applications for identification of inflammatory cells important in plaque formation and rupture. PMID- 16039284 TI - Endurance training increases the number of endothelial progenitor cells in patients with cardiovascular risk and coronary artery disease. AB - OBJECTIVE: As regular physical exercise improves endothelial dysfunction and promotes cardiovascular health, we investigated the effect of training on angiogenesis by measuring the number of circulating endothelial progenitor cells (EPC), the level of EPC-mobilizing growth factors and tested vascular function by flow-mediated dilation (FMD) in patients with coronary artery disease (CAD) and cardiovascular risk factors (CVRF). In addition, degradation products of the NO pathway (NOx) were determined. METHODS AND RESULTS: Twenty patients with documented CAD and/or CVRF joined a 12-week supervised running training. Circulating EPCs--defined by the surface markers CD34, KDR and CD133--were measured at baseline and after exercise training by flow cytometry. We found a significant increase in circulating EPCs (2.9+/-0.4-fold increase; P < .0001), which was positively correlated with both, the change of FMD (r = .81, P < .001) and the increase of NOx synthesis (r = .83, P < .001). Plasma VEGF and erythropoietin did not change in response to exercise. However, we observed a positive correlation between the number of EPCs and erythropoietin at baseline (r = .70, P < .01) and after training (r = .73, P < .01). CONCLUSIONS: Regular exercise training augments the number of circulating EPCs in patients with CVRF and CAD and is associated with improved vascular function and NO synthesis. PMID- 16039285 TI - High frequency of endothelial vasomotor dysfunction after acute coronary syndromes in non-culprit and angiographically normal coronary arteries: a reversible phenomenon. AB - This study aimed to assess endothelium-dependent vasomotor function in non culprit coronary vessels in patients with recent acute coronary syndrome (ACS). Endothelial dysfunction can also concern peripheral vessels during ACS, but the frequency of this phenomenon at coronary circulation level is unknown. Endothelial function was assessed by infusion of graded doses of acetylcholine (ACh) in a non-culprit coronary artery of normal appearance in 43 patients having recently suffered from non-ST ACS, and reassessed 6 months later. Endothelium dependent vasoreactivity was impaired at baseline in 81% (35/43) of the patients, and only 19% (8/43) of patients showed a normal response to ACh. Among the 35 subjects with initial dysfunction, 77% showed a significant improvement 6 months later. All patients without initial endothelial dysfunction remained normal. C reactive protein (CRP) level was elevated at month 0, and had declined at follow up, tending towards normal levels. At that time, apolipoprotein A-I (apoA-I) levels were correlated with vasomotor improvement in univariate (p < 0.02) and multivariate analysis (p < 0.04). In conclusion, endothelium dysfunction is very frequently observed after recent ACS in non-culprit and angiographically normal coronary arteries, and an improvement occurs within 6 months in most cases. After resolution of the initial inflammation, apoA-I seems to play an important role in endothelial function. PMID- 16039286 TI - Remnant lipoproteinemia is a risk factor for endothelial vasomotor dysfunction and coronary artery disease in metabolic syndrome. AB - This study aimed to determine whether elevated levels of remnant lipoprotein, an atherogenic triglyceride-rich lipoprotein, might be associated with coronary artery disease (CAD) and endothelial vasomotor dysfunction in metabolic syndrome. The fasting serum levels of remnant lipoproteins (remnant-like lipoprotein particles cholesterol; RLP-C) were measured by an immunoseparation method in 210 patients with metabolic syndrome meeting ATP III criteria. Flow-mediated endothelium-dependent dilatation (FMD) in the brachial artery during reactive hyperemia was examined by high-resolution ultrasound technique. This study found that elevated RLP-C levels were a significant and independent risk factor for impaired FMD and angiographically proven coronary artery disease (CAD). Treatment with bezafibrate (n = 20) or atorvastatin (n = 20) for 4 weeks significantly reduced RLP-C levels, with a concomitant improvement in FMD. The % reduction in RLP-C levels from baseline after the treatment was independently correlated with the magnitude of improvement in FMD after adjustment for the % changes in levels of triglyceride, hsCRP, and IL-6, and HOMA index. Thus, elevated levels of RLP-C are a risk factor for CAD and endothelial vasomotor dysfunction, a predictor of coronary events, in metabolic syndrome. Measurement of RLP-C is useful for assessment of CAD risk and therapeutic effects in metabolic syndrome. PMID- 16039287 TI - Leukocyte count is associated with reduced endothelial reactivity. AB - BACKGROUND: Leukocyte count has been associated with cardiovascular and cerebrovascular disease in several studies. We hypothesized that white blood cell count is associated with endothelial reactivity. METHODS AND RESULTS: Leukocyte count was measured in a sample of stroke-free community participants undergoing brachial artery testing for endothelial reactivity. Flow-mediated dilation (FMD) during reactive hyperemia was assessed in each subject using high-resolution B mode ultrasound. Multivariate linear regression was used to calculate the effect of leukocyte count on endothelial reactivity after adjusting for potential confounding factors. Mean age of the 868 participants was 66.7+/-8.8 years; 57% were women. Mean leukocyte count was (6.1+/-1.8)x10(9)/L. Each unit increase in leukocyte count was associated with a mean 0.18% decrease in FMD (p = 0.01). After adjusting for other atherosclerosis risk factors, including age, sex, hypertension, diabetes, hyperlipidemia, and smoking, the relationship persisted (mean decrease in FMD per unit leukocyte count = 0.17%, p = 0.02). There was a linear decrease in FMD by quartile of leukocyte count (p = 0.0014). The effect of leukocyte count on FMD was greater for women, those under age 70, and non diabetics. CONCLUSIONS: Relative elevations in leukocyte count are associated with a reduction in brachial artery endothelial reactivity. These findings are consistent with current hypotheses regarding the inflammatory or infectious etiology of risk of atherosclerosis and stroke, but also suggest interactions with demographic and other risk factors. PMID- 16039288 TI - Serum dehydroepiandrosterone sulfate concentration and carotid atherosclerosis in men with type 2 diabetes. AB - Decreased serum dehydroepiandrosterone (DHEA) concentrations may be associated with an increased risk of cardiovascular disease (CVD) in men. We evaluated relationships between serum DHEA sulfate (DHEA-S) concentration and carotid atherosclerosis, as well as major cardiovascular risk factors, in men with type 2 diabetes. Serum DHEA-S concentrations were measured in 206 consecutive men with type 2 diabetes. Relationships were analyzed between serum DHEA-S concentration and carotid atherosclerosis, determined by ultrasonographically evaluated intima media thickness (IMT) and plaque score (PS), as well as major cardiovascular risk factors, including age, blood pressure, and lipid concentrations. Negative correlations were found between DHEA-S concentration and IMT (r = -0.298, P < 0.0001) and between DHEA-S concentration and PS (r = -0.308, P < 0.0001). IMT and PS were significantly greater in patients with lower concentrations of DHEA-S (<1000 ng/ml) than in patients with higher concentrations of DHEA-S (1.07+/-0.30 mm versus 0.91+/-0.19 mm, P < 0.0001, and 5.5+/-4.2 versus 3.1+/-3.4, P < 0.0001, respectively). A negative correlation was found between serum DHEA-S concentration and age (r = -0.488, P < 0.0001). Multiple regression analysis demonstrated that serum DHEA-S concentration was an independent determinant of IMT (beta = -0.289, P < 0.0001) and of PS (beta = -0.301, P < 0.0001). In conclusion, serum DHEA-S concentration is negatively associated with carotid atherosclerosis determined by ultrasonographically evaluated IMT and PS in men with type 2 diabetes. PMID- 16039289 TI - Diurnal and acute patterns of postprandial apolipoprotein B-48 in VLDL, IDL, and LDL from normolipidemic humans. AB - Smaller size particles of intestinal origin may be more relevant to atherosclerosis than larger particles because of easier transendothelial transport. We examined the diurnal pattern in response to low-fat high carbohydrate meals and the effect of a single high-fat milkshake on apoB-48 in very light Sf degrees >400, light Sf degrees 60-400, and dense Sf degrees 20-60 VLDL, IDL, and LDL in 24 subjects (12 in each study). After the high-fat meal, apoB-48 concentrations peaked at 3 h for very light VLDL, 4 h for light VLDL, and 5 h for dense VLDL and IDL, suggesting a delipidation cascade. The diurnal pattern was characterized by a sharp increase in light and dense VLDL apoB-48 after breakfast, reaching the highest concentration 4-8h after breakfast. Less response was observed after lunch, dinner or snack, despite 75% of energy supplied by these meals. The proportion of apoB-48 in LDL in the fasting state ranged between 48 and 54%. LDL apoB-48 concentration decreased by 35% after the high-fat meal, and slowly recovered to near fasting concentrations after 10-12 h. In contrast, in the diurnal study, LDL apoB-48 concentration steady increased peaking 13 h after breakfast, followed by a sharp decrease. In sum, most intestinally synthesized apoB lipoproteins circulate in LDL, and are likely formed in the late postprandial period from larger VLDL-size particles. Although small apoB-48 lipoproteins may be atherogenic remnant particles, their very low concentration may diminish their clinical significance. PMID- 16039290 TI - Adipose tissue triglyceride fatty acids and atherosclerosis in Alaska Natives and non-Natives. AB - Essential polyunsaturated fatty acids (PUFA) of the omega-3 family are believed to protect against cardiovascular disease. A rich source of omega-3 PUFA is found in fish and marine mammals (seal, walrus, whale), which are a large part of the traditional diet of Alaska Natives (Eskimo, American Indians, Aleuts), a group that has been reported to have a lower mortality rate from cardiovascular disease than non-Natives. An autopsy study using standardized methods to evaluate the extent of atherosclerosis and its risk factors, and analyses of stored triglyceride fatty acids was conducted in a sample of Alaska Native subjects and non-Native subjects living in Alaska. Findings indicate that Alaska Natives had less advanced atherosclerosis in coronary arteries, along with higher proportions of omega-3 and lower proportions of omega-6 PUFA in adipose tissue, than did non Natives. We conclude that high dietary intake of omega-3 PUFA may account for the lower extent of coronary artery atherosclerosis, contributing to the reported lower heart disease mortality among Alaska Natives. PMID- 16039291 TI - Cholesterol ester transfer protein (CETP) Taq1B polymorphism influences the effect of a standardized cardiac rehabilitation program on lipid risk markers. AB - Cardiac rehabilitation programs (CR) are standard treatment for patients with coronary artery disease (CAD), yet a large variation in risk factor and lipoprotein changes exists. We investigated the role of three common genetic polymorphisms (CETP Taq1B, LIPC -514 and apo E) associated with alterations of lipoprotein metabolism, in patients before and after standardized CR. Three hundred and seven patients were recruited for this study. DNA samples were collected and all three genotypes were determined for every patient. While the hepatic lipase LIPC promoter polymorphism and apo E genotype showed little or no correlation with response to CR, CETP Taq1B showed significant association with changes in plasma lipid and lipoproteins. The B1 homozygotes for CETP Taq1B genotype showed significant reduction in TC (-0.25+/-0.07, p < 0.01), LDL-C ( 0.15+/-0.06, p < 0.050) and TG (-0.20+/-0.08, p < 0.05). B2 carriers showed no significant change in these parameters. HDL-C, exercise capacity and BMI improved independent of genotype. Individuals with the B1B1 genotype appear to respond well to CR, whereas B2 carriers exhibit marginal gains in lipoprotein risk factors. Although the B2 carriers had similar benefits in exercise capacity and weight reduction, long-term consequences of little or no change in lipoprotein risk factors require further investigation to establish appropriate management strategies. PMID- 16039292 TI - Elevated skin tissue cholesterol levels and myocardial infarction. AB - BACKGROUND: Skin cholesterol has been associated with coronary artery disease, extent of angiographic disease and inflammatory markers such as hs-CRP. Based on these findings we sought to determine whether skin cholesterol was associated with myocardial infarction (MI). METHODS: Patients (N = 649) underwent diagnostic catheterization and concurrent skin cholesterol measurement. History of MI was determined at the time of hospitalization. RESULTS: Patients with a history of MI (n = 225, 35%) had significantly higher skin cholesterol than those without MI (127+/-29 versus 120+/-20, p = 0.002). The odds ratio for high skin cholesterol (for MI) was 1.6 (95% CI = 1.1, 2.6; p = 0.01) after adjustment for traditional risk and extent of angiographic disease. CONCLUSION: Skin cholesterol may indicate increased risk of coronary-related events rather than simply the presence of angiographic narrowing. PMID- 16039293 TI - Long-term effect of estrogen replacement on plasma nitric oxide levels: results from the estrogen in the prevention of atherosclerosis trial (EPAT). AB - The estrogen in the prevention of atherosclerosis trial (EPAT) was a 2-year randomized controlled trial in which unopposed 17beta-estradiol reduced subclinical atherosclerosis progression, measured as change in carotid intima media thickness (CIMT). This study was conducted to determine whether long-term 17beta-estradiol 1mg daily increased plasma nitric oxide (NO) levels and whether this accounted for atheroprotection in EPAT. Although the on-trial serum estradiol level was significantly higher in the estradiol-treated group (n = 91 subjects) than the placebo group (n = 89 subjects) (mean (S.D.) = 59.0 (31.7) pg/ml versus 14.3 (10.4) pg/ml, p < 0.0001), there was no significant difference in the on-trial plasma NO levels, 18.5 (8.2) microM versus 20.1 (9.3) microM. Correlation between on-trial estradiol level and NO change was -0.22 (p = 0.003) in the total sample (placebo- and estradiol-treated subjects) and -0.21 (p = 0.049) in the estradiol-treated group. Change in NO levels was inversely correlated to change in LDL-cholesterol in the estradiol group (r = -0.23, p = 0.03). An NO response to 17beta-estradiol according to age, time since menopause and baseline CIMT was not found arguing against a possible NO effect in healthy versus diseased endothelium. NO levels were not related to CIMT progression. In this study, we found no evidence for an estrogen-induced effect on plasma total NO levels which unlikely accounted for the mechanism underlying the 17beta estradiol atheroprotective effect on subclinical atherosclerosis progression. PMID- 16039294 TI - Association between cigarette smoking, metabolic syndrome, and carotid arteriosclerosis in Japanese individuals. AB - Cigarette smoking is associated with increased insulin resistance and other metabolic abnormalities. Here, we investigate the prevalence of metabolic syndrome (MetS) in cigarette smokers and people who never smoked by analyzing cross-sectional data of 5033 subjects aged between 35 and 65 years who underwent general health screening. Both former and current smoking was associated with an increased incidence of metabolic syndrome defined by modified-National Cholesterol Education Program (NCEP) criteria with odds ratios of 1.77 (95% CI 1.42-2.22, P < 0.0001) and 2.38 (95% CI 1.95-2.91, P < 0.0001), respectively. In both former and current smokers, prevalence of metabolic syndrome increased when the duration of cigarette smoking was > or = 10 years. The positive association between metabolic syndrome and smoking was only partially reversed even 5 years after quitting. Multivariate logistic regression analysis showed that metabolic syndrome was an independent risk factor for carotid plaque with an odds ratio of 1.72 (95% CI 1.43-2.08, P < 0.0001). On the other hand, when limited to individuals without metabolic syndrome, former and current smoking was still found to be associated with carotid plaque with odds ratios of 1.49 (95% CI 1.15 1.92, P = 0.0023) and 1.57 (95% CI 1.22-2.03, P = 0.0005), respectively, in men. Collectively, these data suggest that the atherogenic consequences of smoking may, at least in part, be explained by its association with metabolic syndrome. PMID- 16039295 TI - A genome wide quantitative trait linkage analysis for serum lipids in type 2 diabetes in an African population. AB - Lipid abnormalities are strongly linked with coronary heart disease and are common in type 2 diabetes. However, little is known about the genetic determinants of serum lipids in African populations. An autosomal genome scan was performed for linkage to five plasma lipid phenotypes (total cholesterol, triglycerides (TG), HDL-cholesterol (HDL-C), LDL-cholesterol (LDL-C) and VLDL cholesterol (VLDL-C)) in the Africa-America Diabetes Mellitus (AADM) study. Two hundred and ninety-five affected sibling pairs with type 2 diabetes mellitus enrolled from Ghana and Nigeria were genotyped for 390 microsatellite markers with an average inter-marker distance of 9cM. Multipoint variance components linkage analysis showed that HDL-C had a LOD score of 4.34 near marker D7S3061 and 3.00 near marker D7S513. Some clustering of linkage evidence to several lipid phenotypes was observed on chromosomes 5 (LDL-C, total cholesterol, VLDL-C), chromosome 7 (HDL-C, TG) and chromosome 19 (total cholesterol, LDL-C, TG). Principal component analysis of the five phenotypes yielded two factors, one (TG, HDL-C and VLDL) of which was linked to QTLs on chromosomes 2, 5 and 7, while the other (total cholesterol and LDL-C) was linked to a different set of QTLs on chromosomes 2, 5 and 18. Several of these regions have been reported to be linked to lipids in other studies. Follow up investigations are warranted in view of the central role serum lipids play in the aetiopathogenesis of cardiovascular disease. PMID- 16039296 TI - Beta blocker effects on plasma homocysteine levels in patients with hypertension. AB - Recent studies have shown that hyperhomocysteinemia might play a role in the pathogenesis of hypertension. The effects of antihypertensive agents on plasma homocysteine levels have not been tested extensively. We investigated the effects of beta-blocker therapy on homocysteine levels in patients with hypertension. In the study, 120 patients with newly diagnosed hypertension were enrolled. All patients received metoprolol succinate 100 mg/day initially. If blood pressure was above normal on the 15th day of follow-up, the metoprolol dosage was doubled. Before initiation of the antihypertensive medication and after the fourth month of treatment, homocysteine levels were measured. Of the 120 patients enrolled, 39 could not complete the study. Homocysteine levels decreased significantly by the end of the fourth month when compared with basal values (13.5+/-4.5 micromol/l versus 12.4+/-4.9 micromol/l; P = 0.001). There was no relation between homocysteine level and blood pressure control. There was a significant decrease in homocysteine levels in the women treated in this study (P = 0.001); however, this effect was absent in men (P = 0.185). We demonstrate that metoprolol succinate treatment significantly decreases plasma homocysteine levels in patients with hypertension, especially in women. PMID- 16039297 TI - Postprandial increase of plasma apoAV concentrations in Type 2 diabetic patients. AB - Postprandial hypertriglyceridemia is considered as a risk factor for cardiovascular disease in Type 2 diabetes. However, little is known about the underlying mechanisms. Since the recently discovered apolipoprotein (apo) AV was identified as a modulator of triglyceride (TG) metabolism, the aim of the study was to determine the postprandial apoAV profile of Type 2 diabetic patients. We compared data from 11 patients with Type 2 diabetes mellitus to that of 12 non diabetic normolipidemic subjects following the ingestion of a lipid-rich cream. Postprandial apoAV was elevated in diabetic patients but no correlation was observed either with plasma TG concentration or with the intensity of lipoprotein lipase-dependent lipolysis. These data obtained in human subjects suggest that plasma apoAV concentration does not play an acute or a direct role in the regulation of plasma TG in the postprandial state. PMID- 16039298 TI - Progesterone abolishes estrogen and/or atorvastatin endothelial dependent vasodilatory effects. PMID- 16039300 TI - History of the Society for Invertebrate Pathology. AB - Scientists studying diseases of invertebrates in the USA, Europe, and Asia began to meet at international congresses in the 1950s and early 1960s, and soon recognized that they needed both a society and a journal where their common interests could be discussed and their findings presented. Edward A. Steinhaus played a major role in bringing together scientists from across the globe with common interests in these diseases. As a consequence, the Journal of Invertebrate Pathology (then Journal of Insect Pathology) was initiated in 1959 and Steinhaus became its first editor. Along with Albert Sparks he organized a meeting at Seattle, Washington in 1967 that led to the founding of the Society for Invertebrate Pathology with Steinhaus as its first President. The Society held its first meeting at Ohio State University in 1968, and has continued to meet annually. The Society has instituted a Founder's Lecture series, graduate student awards, and Divisions of Microbial Control, Microsporidia, Bacteriology, Fungi, Viruses, and Nematodes. Members enjoy several social functions at meetings as well as symposia, submitted papers, and poster sessions. The Society for Invertebrate Pathology is a truly international organization which to date has held meetings in 13 countries and 14 US states, usually attended by members from at least 20 countries. PMID- 16039301 TI - Microsporidia and the Society for Invertebrate Pathology [corrected]: a personal point of view. AB - In this overview, I trace the history of the study of microsporidia, with special emphasis on the collegial relationships that developed at the international level and were fostered by the establishment of the Society for Insect Pathology, which later became the Society for Invertebrate Pathology. Study of these organisms of invertebrates in the early days seemed to be mere curiosities, but it soon became clear that they were major disease-causing agents in insects, and later even in vertebrates, especially humans with compromised immune systems. Though microsporidia have not proven effective as pesticides, they do play a role in the regulation of insect populations, especially insects such as the gypsy moth, grasshoppers, and occasionally mosquitoes. PMID- 16039302 TI - From Metchnikoff to Monsanto and beyond: the path of microbial control. AB - In 125 years since Metchnikoff proposed the use of Metarhizium anisopliae to control the wheat cockchafer and brought about the first field trials, microbial control has progressed from the application of naturalists' observations to biotechnology and precision delivery. This review highlights major milestones in its evolution and presents a perspective on its current direction. Fungal pathogens, the most eye-catching agents, dominated the early period, but major mycological control efforts for chinch bugs and citrus pests in the US had questionable success, and interest waned. The discoveries of Bacillus popilliae and Bacillus thuringiensis began the era of practical and commercially viable microbial control. A program to control the Japanese beetle in the US led to the discovery of both B. popilliae and Steinernema glaseri, the first nematode used as a microbial control agent. Viral insect control became practical in the latter half of the 20th century, and the first registration was obtained with the Heliothis nuclear polyhedrosis virus in 1975. Now strategies are shifting for microbial control. While Bt transgenic crops are now planted on millions of hectares, the successes of more narrowly defined microbial control are mainly in small niches. Commercial enthusiasm for traditional microbial control agents has been unsteady in recent years. The prospects of microbial insecticide use on vast areas of major crops are now viewed more realistically. Regulatory constraints, activist resistance, benign and efficacious chemicals, and limited research funding all drive changes in focus. Emphasis is shifting to monitoring, conservation, integration with chemical pesticides, and selection of favorable venues such as organic agriculture and countries that have low costs, mild regulatory climates, modest chemical inputs, and small scale farming. PMID- 16039303 TI - Insecticidal bacteria: an overwhelming success for invertebrate pathology. AB - The discovery and study of insecticidal bacteria, which began a little over a century ago, led to the development of commercial bacterial insecticides in the middle of the century that became the first successful and widely used microbial control agents. Most of these products were based on Bacillus thuringiensis, a bacterium that kills insects through the use of insecticidal proteins that subsequently became known as Cry proteins. While most of these products were only effective against lepidopteran pests, their success eventually led in the 1970s and 1980s to the discovery of strains effective against larvae of coleopteran pests and nematocerous dipterans, such as vector and nuisance mosquitoes and blackflies. The cloning in 1981 of the first gene encoding a Cry protein led to an explosion of basic and applied research that culminated in new strains of recombinant insecticidal bacteria and, even more importantly, the development, commercialization, and wide-scale deployment of insecticidal transgenic crops based on Cry proteins. This new and environmentally safe technology has revolutionized agricultural pest control, yielding a multibillion dollar industry that is paving the way to new types of plants that will dominate food and fiber production as the 21st century progresses. In this brief symposium paper, I provide an overview of some of the key work that led to this remarkable success. PMID- 16039304 TI - A brief journey with insect viruses with emphasis on baculoviruses. PMID- 16039305 TI - The fungal past, present, and future: germination, ramification, and reproduction. PMID- 16039306 TI - Insect-parasitic nematodes: from lab curiosities to model organisms. AB - Interest in studying insect-parasitic nematodes was originally focused on their potential as biological control agents of insects and other arthropod pests. Now, after 30 years of intense basic and applied research, realization of the practical use of insect-parasitic nematodes, particularly of entomopathogenic nematodes and their symbiotic bacteria, has spurred developments across a far broader scientific front. We are now entering a new era of discovery in which tools of molecular genetics are being increasingly used to address a range of biological questions. The knowledge gained from these efforts will directly benefit the practical application of insect-parasitic nematodes as more effective biopesticides. Moreover, these studies will advance these nematodes as unique and intrinsically interesting biological model systems not only for basic research but also in applied fields such as plant health, human medicine, pharmaceutical bioprospecting, and genetic engineering. In this review, the past and current state of insect-parasitic nematode research is summarized. Future research priorities and goals are identified and discussed. PMID- 16039307 TI - Observations on the history of non-insect invertebrate pathology from the perspective of a participant. AB - This paper discusses diseases of commercially important non-insect invertebrates from both a historical and a personal perspective. Early studies of curious scientists such as Metchnikoff using invertebrate systems led to important discoveries with much wider impact. As commercial culture of oysters, shrimp, and other invertebrates grew, so did studies of their diseases. Here, the impact of pioneering scientists and important new techniques on our understanding of these diseases is described. PMID- 16039308 TI - The Oryctes virus: its detection, identification, and implementation in biological control of the coconut palm rhinoceros beetle, Oryctes rhinoceros (Coleoptera: Scarabaeidae). AB - In view of the increasing and devastating damage by rhinoceros beetle (Oryctes rhinoceros) to coconut palms in the middle of last century, many efforts were made to find an efficient natural control factor against this pest, which could not be controlled by pesticides. The basic procedures of these monitoring programmes are outlined together with the final detection of a virus disease in oil palm estates in Malaysia in 1963. In extensive laboratory studies, the virus was isolated and identified as the first non-occluded, rod-shaped insect virus, morphologically resembling the baculoviruses. Infection experiments clarified the pathology, histopathology, and virulence of the virus and demonstrated that the virus was extremely virulent to larvae after peroral application. These findings encouraged the first pilot release of virus in 1967 in coconut plantations of Western Samoa where breeding sites were contaminated with virus. Surprisingly, the virus became established in the Samoan rhinoceros beetle populations and spread autonomously throughout the Western Samoan islands. As a consequence, there was a drastic decline of the beetle populations followed by a conspicuous recovery of the badly damaged coconut stands. This unexpected phenomenon could only be explained after it was shown that the adult beetle itself is a very active virus vector and thus was responsible for the efficient autodissemination of the virus. The functioning of the beetle as a 'flying virus factory' is due to the unique cytopathic process developing in the midgut after peroral virus infection. Pathological details of this process are presented. Because of the long-term persistence of the virus in the populations, rhinoceros beetle control is maintained. Incorporation of virus into integrated control measures and successful virus releases in many other countries are recorded. PMID- 16039309 TI - The incidence and use of Oryctes virus for control of rhinoceros beetle in oil palm plantations in Malaysia. AB - The rhinoceros beetle, Oryctes rhinoceros, has emerged as a serious pest of oil palm since the prohibition of burning as a method for maintaining estate hygiene in the 1990s. The abundance of beetles is surprising given that the Malay peninsula was the site of first discovery of the Oryctes virus, which has been used to effect good as a biological control agent in other regions. A survey of adult beetles was carried out throughout Malaysia using pheromone traps. Captured beetles were examined for presence of virus using both visual/microscopic examination and PCR detection methods. The survey indicated that Oryctes virus was common in Malaysia among the adult beetles. Viral DNA analysis was carried out after restriction with HindIII enzyme and indicated at least three distinct viral genotypes. Bioassays were used to compare the viral strains and demonstrate that one strain (type B) is the most virulent against both larvae and adults of the beetle. Virus type B has been cultured and released into healthy populations where another strain (type A) forms the natural background. Capture and examination of beetles from the release site and surrounding area has shown that the spread and persistence of the applied virus strain is accompanied by a reduction in palm frond damage. PMID- 16039310 TI - Oryctes virus--time for a new look at a useful biocontrol agent. AB - The introduction of Oryctes virus into outbreak areas of the rhinoceros beetle, Oryctes rhinoceros (Coleoptera: Scarabaeidae), has been a major success for "classical" biocontrol with a virus and has led to a dramatic reduction in palm damage in many areas of the Asia/Pacific region. In recent years, however, there have been new reports of high levels of rhinoceros beetle damage to palms. Damage has been especially intense in SE Asia following the introduction of no-burn polices for land clearance and replanting, but outbreaks have also been reported from some Pacific Islands where control seems to have diminished over time. SE Asian studies show that there is considerable genetic variation among endemic Oryctes virus isolates and studies in new island release areas have shown rapid evolution of the virus. The consequences of such genetic variation are in need of further study. Furthermore, the taxonomic position of the virus is unclear, with its removal from the Baculoviridae to an "unassigned' virus, reflecting its novel characteristics. Genomic sequencing could help resolve the taxonomy of the virus and provide a basis for studying strain variation. Oryctes virus has achieved wide success in the past without the benefit of molecular analysis and identification techniques. In order to fully take advantage of this unique pathogen for protection of palms, a renewed, coordinated effort centered on genetic selection and distribution of effective strains is required. PMID- 16039311 TI - Uterine leiomyosarcomas express KIT protein but lack mutation(s) in exon 9 of c KIT. PMID- 16039312 TI - Re: "P63 and EGFR as prognostic predictors in stage IIB radiation-treated cervical squamous cell carcinoma". PMID- 16039313 TI - Adjuvant carboplatin in stage I seminoma. PMID- 16039314 TI - Physical activity and obesity in childhood and adolescence. PMID- 16039315 TI - Screening for haemochromatosis--producing or preventing illness? PMID- 16039316 TI - Being realistic about benefits of supervised injecting facilities. PMID- 16039317 TI - High-dose chemotherapy and allogeneic immunotherapy. PMID- 16039318 TI - Undue inducement in clinical research. PMID- 16039319 TI - A dismal and dangerous verdict against Roy Meadow. PMID- 16039321 TI - Sterilisation. PMID- 16039322 TI - In the days of patients' choice, why is the patient being ignored? PMID- 16039323 TI - A tax to prevent the epidemic of lung cancer. PMID- 16039324 TI - A tax to prevent the epidemic of lung cancer. PMID- 16039325 TI - Aseptic peritonitis due to peptidoglycan. PMID- 16039327 TI - Emergency contraception. PMID- 16039329 TI - MARIBS study. PMID- 16039328 TI - HIV/AIDS in South Africa. PMID- 16039330 TI - The vote against the European Constitution. PMID- 16039331 TI - Radiotherapy versus single-dose carboplatin in adjuvant treatment of stage I seminoma: a randomised trial. AB - BACKGROUND: Adjuvant radiotherapy is effective treatment for stage I seminoma, but is associated with a risk of late non-germ-cell cancer and cardiovascular events. After good results in initial studies with one injection of carboplatin, we undertook a large randomised trial to compare the approaches of radiotherapy with chemotherapy in seminoma treatment. METHODS: Between 1996 and 2001, 1477 patients from 70 hospitals in 14 countries were randomly assigned to receive radiotherapy (para-aortic strip or dog-leg field; n=904) or one injection of carboplatin (n=573; dose based on the formula 7x[glomerular filtration rate+25] mg), at two trial centres in the UK and Belgium. The primary outcome measure was the relapse-free rate, with the trial powered to exclude absolute differences in 2-year rates of more than 3%. Analysis was by intention to treat and per protocol. This trial has been assigned the International Standard Randomised Controlled Trial Number ISRCTN27163214. FINDINGS: 885 and 560 patients received radiotherapy and carboplatin, respectively. With a median follow-up of 4 years (IQR 3.0-4.9), relapse-free survival rates for radiotherapy and carboplatin were similar (96.7% [95% CI 95.3-97.7] vs 97.7% [96.0-98.6] at 2 years; 95.9% [94.4 97.1] vs 94.8% [92.5-96.4] at 3 years, respectively; hazard ratio 1.28 [90% CI 0.85-1.93], p=0.32). At 2 years' follow-up, the absolute differences in relapse free rates (radiotherapy-chemotherapy) were -1.0% (90% CI -2.5 to 0.5) by direct comparison of proportions, and 0.9% (-0.5 to 3.0) by a hazard-ratio-based approach. Patients given carboplatin were less lethargic and less likely to take time off work than those given radiotherapy. New, second primary testicular germ cell tumours were reported in ten patients allocated irradiation (all after para aortic strip field) and two allocated carboplatin (5-year event rate 1.96% [95% CI 1.0-3.8] vs 0.54% [0.1-2.1], p=0.04). One seminoma-related death occurred after radiotherapy and none after carboplatin. INTERPRETATION: This trial has shown the non-inferiority of carboplatin to radiotherapy in the treatment of stage I seminoma. Although the absence of disease-related deaths and preliminary data indicating fewer second primary testicular germ-cell tumours favour carboplatin use, these findings need to be confirmed beyond 4 years' follow-up. PMID- 16039332 TI - Relation between the changes in physical activity and body-mass index during adolescence: a multicentre longitudinal study. AB - BACKGROUND: The role of physical activity in preventing obesity during adolescence remains unknown. We examined changes in activity in relation to changes in body-mass index (BMI) and adiposity in a cohort of 1152 black and 1135 white girls from the USA, who were followed up prospectively from ages 9 or 10 to 18 or 19 years. METHODS: BMI and sum of skinfold thickness were assessed annually, whereas habitual activity was assessed at years 1 (baseline), 3, 5, and 7-10. Each girls' overall activity status was categorised as active, moderately active, or inactive. Longitudinal regression models examined associations between changes in activity and in overall activity status with changes in BMI and in sum of skinfold thickness. FINDINGS: Each decline in activity of 10 metabolic equivalent [MET]-times per week was associated with an increase in BMI of 0.14 kg/m2 (SE 0.03) and in sum of skinfold thickness of 0.62 mm (0.17) for black girls, and of 0.09 kg/m2 (0.02) and 0.63 mm (0.13) for white girls. At ages 18 or 19 years, BMI differences between active and inactive girls were 2.98 kg/m2 (p<0.0001) for black girls and 2.10 kg/m2 (p<0.0001) for white girls. Similar results were apparent for sum of skinfold thickness. For moderately active girls, changes in BMI and sum of skinfold thickness were about midway between those for active and inactive girls. INTERPRETATION: Changes in activity levels of US girls during adolescence significantly affected changes in BMI and adiposity. Thus, preventing the steep decline in activity during adolescence is an important method to reduce obesity. PMID- 16039333 TI - Ceftriaxone as effective as long-acting chloramphenicol in short-course treatment of meningococcal meningitis during epidemics: a randomised non-inferiority study. AB - BACKGROUND: In sub-Saharan Africa in the 1990s, more than 600,000 people had epidemic meningococcal meningitis, of whom 10% died. The current recommended treatment by WHO is short-course long-acting oily chloramphenicol. Continuation of the production of this drug is uncertain, so simple alternatives need to be found. We assessed whether the efficacy of single-dose treatment of ceftriaxone was non-inferior to that of oily chloramphenicol for epidemic meningococcal meningitis. METHODS: In 2003, we undertook a randomised, open-label, non inferiority trial in nine health-care facilities in Niger. Participants with suspected disease who were older than 2 months were randomly assigned to receive either chloramphenicol or ceftriaxone. Primary outcome was treatment failure (defined as death or clinical failure) at 72 h, measured with intention-to-treat and per-protocol analyses. FINDINGS: Of 510 individuals with suspected disease, 247 received ceftriaxone, 256 received chloramphenicol, and seven were lost to follow-up. The treatment failure rate at 72 h for the intention-to-treat analysis was 9% (22 patients) for both drug groups (risk difference 0.3%, 90% CI -3.8 to 4.5). Case fatality rates and clinical failure rates were equivalent in both treatment groups (14 [6%] ceftriaxone vs 12 [5%] chloramphenicol). Results were also similar for both treatment groups in individuals with confirmed meningitis caused by Neisseria meningitidis. No adverse side-effects were reported. INTERPRETATION: Single-dose ceftriaxone provides an alternative treatment for epidemic meningococcal meningitis--its efficacy, ease of use, and low cost favour its use. National and international health partners should consider ceftriaxone as an alternative first-line treatment to chloramphenicol for epidemic meningococcal meningitis. PMID- 16039334 TI - Use of community genetic screening to prevent HFE-associated hereditary haemochromatosis. AB - HFE-associated hereditary haemochromatosis is a recessive, iron-overload disorder that affects about one in 200 north Europeans and that can be easily prevented. However, genetic screening for this disease is controversial, and so we assessed whether such screening was suitable for communities. Cheek-brush screening for the Cys282Tyr HFE mutation was offered to individuals in the workplace. Outcomes were assessed by questionnaires before and after testing. 11,307 individuals were screened. We recorded no increase in anxiety in individuals who were homozygous for the Cys282Tyr mutation or non-homozygous. Self-reported tiredness before testing was significantly higher in homozygous participants than in non homozygous participants (chi2 test, p=0.029). Of the 47 homozygous individuals identified, 46 have taken steps to treat or prevent iron accumulation. Population genetic screening for HFE-associated hereditary haemochromatosis can be practicable and acceptable. PMID- 16039335 TI - Safer injection facility use and syringe sharing in injection drug users. AB - Safer injection facilities provide medical supervision for illicit drug injections. We aimed to examine factors associated with syringe sharing in a community-recruited cohort of illicit injection drug users in a setting where such a facility had recently opened. Between Dec 1, 2003, and June 1, 2004, of 431 active injection drug users 49 (11.4%, 95% CI 8.5-14.3) reported syringe sharing in the past 6 months. In logistic regression analyses, use of the facility was independently associated with reduced syringe sharing (adjusted odds ratio 0.30, 0.11-0.82, p=0.02) after adjustment for relevant sociodemographic and drug-use characteristics. These findings could help inform discussions about the merits of such facilities. PMID- 16039336 TI - Reduced intensity conditioning for allograft after cytoreductive autograft in metastatic breast cancer. AB - The benefits of allografting noted in some malignant diseases might be safely extended to metastatic breast cancer by a combination of cytoreduction with high dose chemotherapy (HDT) and autologous stem-cell transplant (ASCT) with graft versus-tumour effect mediated by transplanted donor immune cells with nonmyeloablative allografting (reduced intensity conditioning transplantation, RICT). 17 patients with heavily pretreated disease were given tandem transplants. 13 patients sustained donor engraftment. Three had partial remission after HDT and ASCT and complete remission after RICT; they achieved full chimerism and all developed graft-versus-host disease (GVHD) before regression of cancer. Another patient did not respond to HDT and ASCT but had partial remission after RICT, giving an overall response rate of 24%. Five patients had grade II or higher acute GVHD and five had extensive chronic GVHD. No non-relapse-related deaths occurred during the first 100 days. Five patients (29%) were alive 90-2160 days (median 1320) after RICT. This two-step approach is feasible in patients with metastatic breast cancer. PMID- 16039337 TI - Sjogren's syndrome. AB - Sjogren's syndrome is a chronic autoimmune disorder of the exocrine glands with associated lymphocytic infiltrates of the affected glands. Dryness of the mouth and eyes results from involvement of the salivary and lacrimal glands. The accessibility of these glands to biopsy enables study of the molecular biology of a tissue-specific autoimmune process. The exocrinopathy can be encountered alone (primary Sjogren's syndrome) or in the presence of another autoimmune disorder such as rheumatoid arthritis, systemic lupus erythematosus, or progressive systemic sclerosis. A new international consensus for diagnosis requires objective signs and symptoms of dryness including a characteristic appearance of a biopsy sample from a minor salivary gland or autoantibody such as anti-SS-A. Exclusions to the diagnosis include infections with HIV, human T-lymphotropic virus type I, or hepatitis C virus. Therapy includes topical agents to improve moisture and decrease inflammation. Systemic therapy includes steroidal and non steroidal anti-inflammatory agents, disease-modifying agents, and cytotoxic agents to address the extraglandular manifestations involving skin, lung, heart, kidneys, and nervous system (peripheral and central) and haematological and lymphoproliferative disorders. The most difficult challenge in diagnosis and therapy is patients with symptoms of fibromyalgia (arthralgia, myalgia, fatigue) and oral and ocular dryness in the presence of circulating antinuclear antibodies. PMID- 16039338 TI - King George III and porphyria: an elemental hypothesis and investigation. AB - In 1969 it was proposed that the episodic madness suffered by King George III (1738-1820) resulted from an acute hereditary porphyria, variegate porphyria, caused by deficiency of protoporphyrinogen oxidase. The diagnosis was based on the historical archive and a contentious claim that living members of the House of Hanover were affected with the condition. A re-examination of the medical evidence and the appearance of new historical material have suggested that porphyria did indeed exist in the Royal Houses of Europe. We report the analysis of hair obtained from George III. Although no genomic DNA could be obtained, metal analysis revealed high concentrations of arsenic. Since arsenic interferes with haem metabolism, it might have contributed to the King's unusually severe and prolonged bouts of illness. We have identified sources of arsenic in the context of the medication George III received from physicians. PMID- 16039339 TI - Undue inducement in clinical research in developing countries: is it a worry? PMID- 16039340 TI - Mesh mimics metastasis. PMID- 16039341 TI - Radioisotopes and airport security. PMID- 16039342 TI - Applications for release by Australians in Victoria found not guilty of offences of violence by reason of mental impairment. AB - Much research in relation to mental illness and the law has concentrated upon when accused persons are entitled to avail themselves of the defence of not guilty by reason of insanity or mental impairment. However, the decision as to when persons found not guilty by such pleas should be released step by step back into the community involves difficult analyses of the risk of recidivism by persons who have committed serious acts of violence whilst mentally ill. This article analyses some 70 cases heard by the Supreme Court of Victoria in Australia since the jurisdiction to make such decisions has been transferred from the executive arm of government to the judiciary. The jurisprudence generated by the Victorian Supreme Court constitutes Australia's most developed law in relation to prediction of dangerousness. This article evaluates the different and subtle dynamics that have influenced the judges in an increasingly sophisticated way to grapple with the phenomenon of mental illness in deciding when persons who have already killed can safely be released from involuntary detention status within the confines of a forensic psychiatric institution back into the general community. PMID- 16039343 TI - Public participation in regional health policy: a theoretical framework. AB - How best to involve the public in local health policy development and decision making is an ongoing challenge for health systems. In the current literature on this topic, there is discussion of the lack of rigorous evaluations upon which to draw generalizable conclusions about what public participation methods work best and for what kinds of outcomes. We believe that for evaluation research on public participation to build generalizable claims, some consistency in theoretical framework is needed. A major objective of the research reported on here was to develop such a theoretical framework for understanding public participation in the context of regionalized health governance. The overall research design followed the grounded theory tradition, and included five case studies of public participation initiatives in an urban regional health authority in Canada, as well as a postal survey of community organizations. This particular article describes the theoretical framework developed, with an emphasis on explaining the following major components of the framework: public participation initiatives as a process; policy making processes with a health region; social context as symbolic and political institutions; policy communities; and health of the population as the ultimate outcome of public participation. We believe that this framework is a good beginning to making more explicit the factors that may be considered when evaluating both the processes and outcomes of public participation in health policy development. PMID- 16039344 TI - Using a population-based database to explore the inter-specialty differences in physician practice incomes in Taiwan. AB - Gaining an understanding of the distribution of physician incomes between different medical specialties could assist policymakers to predict the future medical manpower supply. The purpose of this study is to examine the differences in medical specialty-specific gross practice incomes between office-based physicians in Taiwan. The primary data source for the study, which includes 7444 office-based physicians, was provided by the Taiwan Department of Health, with the dependent variable of interest to this study being the annual gross income of physician practices, whilst the independent variable is physician specialty. The study controlled for physicians' age, gender, specialty-board status, type of practice, location of clinic and urbanization level of the community in which the practice was located. Multivariate regression analyses were carried out to explore the relationship between physician specialty and gross practice income. This study finds a significant relationship between the annual gross income of physician practices and the physician's medical specialty (P < 0.001). Of all physicians, those specializing in rehabilitation and orthopedics had the highest gross practice incomes; conversely, obstetricians and gynecologists had the lowest gross practice incomes. The regression analyses demonstrated that after adjusting for socio-demographic and professional characteristics, gross practice incomes of physicians were significantly related to their medical specialty. This study concludes that differences in the gross practice incomes of physicians were significantly related to medical specialties. Those physicians specializing in procedure-based specialties, such as rehabilitation and orthopedics, had higher practice incomes than their counterparts in other more diagnosis-oriented specialties such as family practice and pediatrics. PMID- 16039345 TI - Field based evidence of enhanced healthcare utilization among persons insured by micro health insurance units in Philippines. AB - Underutilization of healthcare is common among rural and low-income population segments in countries with lower income or inequitable income distribution. Micro health insurance units (MIUs) are created by informal sector groups because people cannot access health insurance or are dissatisfied with the programmes they can access. The policy choice to support MIUs relies on evidence that affiliation with these schemes increases healthcare utilization. This article examines new evidence of the association between affiliation with MIUs and healthcare utilization. We analyzed field data collected in 6 MIUs in the Philippines in 2002 (through a household survey encompassing 890 insured- and 1063 uninsured households). The two cohorts did not differ in demographic parameters, and differed only marginally in income and education levels, both higher amongst the insured. Insured persons reported higher hospitalization rates, higher rates of professionally-attended deliveries, lower rates of delivery at home, a higher frequency of primary-care physician encounters, a higher rate of diagnosed chronic diseases, and better drug compliance among chronically ill. Increased utilization by the insured is not due to adverse selection, judging by two facts: morbidity of the two cohorts, as assessed by a proxy indicator (the reported number of episodes of illness) did not differ; and rates of deliveries were even slightly higher among the uninsured. We conclude that MIUs in the Philippines can alleviate underutilization of heath care. PMID- 16039346 TI - Is universal coverage a solution for disparities in health care? Findings from three low-income provinces of Thailand. AB - The policy on universal coverage (UC) of health care has been adopted and implemented incrementally by the government of Thailand since April 2001 with the aim of providing the access to care for the uninsured population. The success of UC, however, depends on how effective its design and implementation arrangements are in reaching population and affecting households' health seeking behavior and abilities to take up benefits of UC. The results from the household survey of 1834 respondents conducted in three low-income provinces (Tak, Sakol Nakorn, Narathiwat) show that the Gold card with exemption scheme was pro-poor while other insurance schemes tended to favor the rich with 2.6% of respondents reported having more than one type of health insurance coverage and 8.9% without health insurance. The insurance status had statistically significant association with health care use, and knowledge on family planning method and sexually transmitted diseases. Additionally, consumer preferences and socioeconomics factors are a key to disparities in health care utilization. PMID- 16039347 TI - Intersectoral problems in the Russian organisation of public health. AB - In spite of the ongoing transition of the Russian society, there is still a traditional view of public health, which is based to a great extent on the ideals and priorities of the Soviet period. Public health activities are regarded mainly as a responsibility of the health sector. There are, however, important public health activities going on also in other sectors of the society, for example, in the educational sector and the local communities, but also in the social insurance system. There is an important Russian tradition of prophylactic treatment in sanatoriums and health resorts, which is financed to a large extent by the social insurance. Based on three qualitative empirical studies, this article describes the organisation of public health in the Russian Federation and analyses the problems of intersectoral co-ordination and collaboration within this organisation. The analysis is focusing on the relations between the health sector and the social insurance system, which are not so well known outside the country. The results of this analysis show a fragmented organisation with a serious lack of co-ordination, but also a limited collaboration between the different sectors involved in public health. On the basis of these results, there is a discussion of how intersectoral collaboration could be improved in the Russian organisation of public health. PMID- 16039348 TI - Message in a bottle: sinking in a sea of safe motherhood concepts. AB - The experiences of implementing maternal health programmes over the last two decades have resulted in the development of many approaches and concepts to address the problems of maternal death and disability in developing countries. These safe motherhood "messages" are generally conveyed from international organisations to implementers of programmes working in developing countries. The messages are sometimes unclear, ambiguous and open to misinterpretation. Case studies are used to describe varying interpretations of messages on essential and emergency obstetric care, skilled attendance at delivery and measurement of progress. Limited technological access to information, rapidly changing ideology, overly complicated terminology, inadequate evidence, poor international and inter agency consensus are key reasons contributing to confusion in implementation. Policy-implementation gaps can be bridged with better needs-based evidence, improved consistency and means of delivery of global messages, building capacity, strengthening partnerships and more inclusive participation in the global arena. PMID- 16039349 TI - The costs of interpersonal violence--an international review. AB - This article reviews evidence of the economic impact of interpersonal violence internationally. In the United States, estimates of the costs of interpersonal violence reach 3.3% of GDP. The public sector-and thus society in general-bears the majority of these costs. Interpersonal violence is defined to include violence between family members and intimate partners, and violence between acquaintances and strangers that is not intended to further the aims of any formally defined group or cause. Although these types of violence disproportionately affect poorer countries, there is a scarcity of studies of their economic impact in these countries. International comparisons are complicated by the calculation of economic losses based on foregone wages and income, thus undervaluing economic losses in poorer countries. PMID- 16039350 TI - The decline in 'free' general practitioner care in Australia: reasons and repercussions. AB - The Australian public insurer, Medicare, allows general practitioners (GPs) to bulk bill patients, or accept the government rebate as full payment for their services. The percentage of GP consultations bulk billed, however has declined from 78.6% in June 2000 to 65.7% in December 2003. The immediate impact of a declining level of bulk billing is a decrease in the availability of free GP health care for patients. This has implications for copayments and access to GPs for low income groups in particular. In this paper, we explore the reasons for and repercussions of the decline in bulk billing. We analyse two main reasons for the decline. The first is a failure of the rebate to maintain a level consistent with increases in medical practice costs. The second is a decline in GPs in some regional and rural areas resulting in a decrease in price competition. The government has recently made changes to deal with the decline in bulk billing and based on three quarters of data, there has been a modest improvement in bulk billing. PMID- 16039351 TI - Reimbursement decisions in health policy--extending our understanding of the elements of decision-making. AB - Previous theoretical and empirical work on health policy decisions about reimbursement focuses on specific rationales such as effectiveness, economic considerations and equal access for equal needs. As reimbursement decisions take place in a social and political context we propose that the analysis of decision making should incorporate factors, which go beyond those commonly discussed. As an example we chose three health technologies (sildenafil, rivastigmine and statins) to investigate how decisions about reimbursement of medicines are made in the United Kingdom National Health Service and what factors influence these decisions. From face-to-face, in-depth interviews with a purposive sample of 20 regional and national policy makers and stakeholders we identified two dimensions of decision-making, which extend beyond the rationales conventionally cited. The first dimension relates to the role of 'subjectivity' or 'the personal' in the decisions, including personal experiences of the condition and excitement about the novelty or potential benefit of the technology-these factors affect what counts as evidence, or how evidence is interpreted, in practice. The second dimension relates to the social and political function of decision-making and broadens what counts as the relevant ends of decision-making to include such things as maintaining relationships, avoiding organisational burden, generating politically and legally defensible decisions and demonstrating the willingness to care. More importantly, we will argue that these factors should not be treated as contaminants of an otherwise rational decision-making. On the contrary we suggest that they seem relevant, reasonable and also of substantial importance in considering in decision-making. Complementing the analysis of decision-making about reimbursement by incorporating these factors could increase our understanding and potentially improve decision-making. PMID- 16039352 TI - Are the health Millennium Development Goals appropriate for Eastern Europe and Central Asia? AB - This article argues that the health-related Millennium Development Goals do not appropriately address the challenges faced by the countries of Eastern Europe and Central Asia. By ignoring adult mortality, their achievement would result in relatively small gains in life expectancy. To achieve greater impact, policies in this region must supplement the classical Millennium Development Goals with indicators of adult health, in particular cardiovascular diseases and external causes of death. In addition, countries, with support from the international community, must improve the quality of vital registration data to enable more accurate estimation of the disease burden. PMID- 16039353 TI - A qualitative assessment of rheumatoid hand surgery in various regions of the world. AB - Differences in opinion between rheumatologists and hand surgeons result in disparate practice patterns among different states in the United States regarding the use of surgery for hand deformities in the rheumatoid population. The purpose of this project was to study the practices of rheumatoid hand surgery from countries in various regions of the world. PMID- 16039354 TI - Long-term follow-up study of radiocarpal arthrodesis for the rheumatoid wrist. AB - PURPOSE: Pain-free stability of the wrist is a prerequisite for the rheumatoid hand to maintain power and perform various tasks. The purpose of this study was to investigate whether a radiocarpal (radiolunate, radioscapholunate, or radiolunotriquetral) arthrodesis produces a stable wrist and whether the results remain satisfactory for more than 10 years. METHODS: A retrospective review was performed on 25 wrists of 25 patients with rheumatoid arthritis who had radiocarpal arthrodesis. All patients had a synovectomy of the extensor tendons and the wrist joint combined with a Darrach procedure. The indications for radiocarpal arthrodesis included radiographic changes in Larsen-Dale-Eek grades II to IV, midcarpal joint space of greater than 1 mm, and ulnar shift or palmar subluxation of the carpus. The presence of scapholunate dissociation was an optional indication. The mean follow-up period was 13 years (range, 10-18 y) and radiographs taken just before the surgery and 0 to 2 years, 2 to 5 years, 5 to 10 years, and more than 10 years after the surgery were evaluated as were pain relief, swelling, grip power, range of motion, and complications. RESULTS: Pain was resolved for 22 of the patients and 3 experienced occasional mild pain. Swelling generally decreased, grip power increased significantly, flexion decreased, and forearm rotation increased significantly. The complication rate was low. Radiographically ulnar shift and palmar subluxation improved initially and were maintained at the time of the 10-year follow-up evaluation; carpal collapse improved initially but returned to the preoperative level by the time of the 5-year follow-up evaluation. The midcarpal joint space was preserved in 16 wrists, and all but 1 wrist (in a patient with mutilating type of the disease) remained stable. CONCLUSIONS: Radiocarpal arthrodesis for treatment of the rheumatoid wrist results in good stability with preservation of motion despite radiographic progression of the disease. We therefore recommend this treatment for the unstable wrist with moderate deterioration. PMID- 16039355 TI - Silicone implant arthroplasty in patients with idiopathic osteoarthritis of the metacarpophalangeal joint. AB - PURPOSE: The outcome of silicone metacarpophalangeal (MCP) joint implant arthroplasty in the osteoarthritic patient population has not been well established. Typically patients with idiopathic osteoarthritis have no history of underlying systemic disease and may respond well to treatment with MCP joint implant arthroplasty. This study examined the efficacy of silicone MCP joint implant arthroplasty for patients with idiopathic osteoarthritis for whom nonsurgical treatment had failed. METHODS: Of 14 patients (15 arthroplasties) who had silicone MCP joint implant arthroplasty for idiopathic osteoarthritis 12 (13 arthroplasties) returned for follow-up evaluation at an average of 40 months after surgery. There were 9 index finger and 4 middle finger arthroplasties. The average age at the time of surgery was 62 years. Patients completed a subjective questionnaire and were examined by a certified therapist. Range of motion and strength were recorded and the Jebsen-Taylor examination was administered to assess function. Range of motion values at final follow-up evaluation were compared with preoperative and early postoperative values. Radiographs were taken at final follow-up evaluation and compared with preoperative and early postoperative films to assess joint position, wear, and radioulnar alignment. RESULTS: At final follow-up evaluation excellent (9 patients) and good (3 patients) overall improvement were reported. Nine patients (10 implants) reported greater than 75% functional improvement. A notable increase was seen in MCP joint flexion. Grip and lateral pinch strengths were below age-matched normative data. Of the 11 patients (12 implants) who came in for follow-up evaluation 7 performed all tasks of the Jebsen-Taylor examination within the allotted time. At final follow-up evaluation all silicone implants were located and showed no signs of subluxation. Radiographic radioulnar alignment was maintained. One implant was revised at 35 months secondary to fracture. CONCLUSIONS: Silicone implant arthroplasty is a motion-sparing procedure that provides good pain relief and maintenance of function at intermediate follow-up evaluation in patients with idiopathic osteoarthritis of the MCP joint. PMID- 16039356 TI - Bilateral arthroscopic tendon interposition arthroplasty of the thumb carpometacarpal joint in a patient with Ehlers-Danlos syndrome: a case report. AB - A common finding in progressive osteoarthritis of the carpometacarpal joint of the thumb is ligamentous laxity. In patients with Ehlers-Danlos syndrome, which is a disorder characterized by generalized ligamentous laxity, splinting and arthrodesis have been recommended because of the random results achieved by other reconstructive procedures. We report a patient with thumb carpometacarpal joint osteoarthritis secondary to Ehlers-Danlos syndrome who was treated with bilateral arthroscopic tendon interposition arthroplasty. PMID- 16039357 TI - Lunate resection and vascularized Os pisiform transfer in Kienbock's Disease: an average of 10 years of follow-up study after Saffar's procedure. AB - PURPOSE: To investigate the long-term results of lunate replacement by vascularized bone transfer in advanced Kienbock's disease. METHODS: Twenty-one patients were reviewed (mean follow-up period +/- SD, 9.9 +/- 3.5 y) to analyze results after lunate replacement by vascularized pisiform transposition (Saffar's procedure) for Lichtman stages III and IV. RESULTS: Pain was improved in 16 of 21 patients but range of motion did not improve after surgery. Range of motion was reduced to 68% and grip power to 80% of that of the opposite hand. At follow-up evaluation the mean score on the Disabilities of the Arm, Shoulder, and Hand Questionnaire was 22.3 +/- 17.9 and the mean Cooney score was 75.4 +/- 13.2. Radiologically, Lichtman stage persisted in 8, improved in 1, progressed in 8, and could not be evaluated in 3 patients. Two patients had radiologic signs of arthritis before surgery. At follow-up examination osteoarthritis was found in 50% of patients. The majority of degenerative changes were associated with carpal collapse. CONCLUSIONS: The replacement of the lunate by vascularized pisiform transposition maintained preoperative ranges of motion. At follow-up examination both patient satisfaction and wrist function were high. In the long term, however, Saffar's procedure can restore alignment of the carpus only partly and also results in osteoarthritis in half of all patients. PMID- 16039358 TI - Three-dimensional description of ligamentous attachments around the lunate. AB - PURPOSE: To detail the location and area of specific ligament attachments on the 3-dimensional (3-D) surface morphology of the individual carpal bones. METHODS: We describe the attachment regions of both extrinsic and intrinsic intercarpal ligaments by using a combination of detailed dissection, computed tomography imaging, and a 3-D digitization technique and provide detailed information about ligament attachments around the lunate. RESULTS: The length, width, and thickness of each ligament are described in millimeters and its area of attachment is given in square millimeters and as a percentage of the bone surface and/or total ligament attachment area. The attachment sites of the ligaments also are shown visually on 3-D images of the bones. CONCLUSIONS: This study improves the knowledge and understanding of carpal ligament anatomy, which can result in better intraoperative assessment of ligament integrity/disruption and allow more accurate and anatomic repair, reattachment, and/or reconstruction of carpal ligaments for the treatment of carpal instability. PMID- 16039359 TI - Lunate osteochondroma: a case report. AB - We report a case of osteochondroma of the lunate. A 16-year-old boy had a 6-year history of irritable snapping of the right wrist associated with numbness of the ipsilateral middle finger. At surgery we found that the median nerve became caught on the lunate osteochondroma while the wrist was extended and slipped over it when the wrist was flexed rapidly, thereby causing a snapping phenomenon. The symptoms disappeared completely after the protruding lesion was excised. Several cases of solitary carpal osteochondroma have been reported previously. PMID- 16039360 TI - Both scanning plane and observer affect measurements of scaphoid deformity. AB - PURPOSE: The influence of angular deformity of the scaphoid on wrist function and arthrosis is debated and the reliability of the described quantitative measurements of deformity has been questioned. We hypothesized that the inherent imprecision with which computed tomography scanning planes are selected introduces another source of variability in measurements of scaphoid deformity, further diminishing their reliability. METHODS: Sagittal plane images of 15 computed tomograms of normal scaphoids were evaluated in 3 different reconstruction planes. Four observers measured the lateral intrascaphoid angle, the dorsal cortical angle, and the height-to-length ratio of the 45 images in random order and then measured them again in a distinct random order 2 weeks later. The variability of each observer's measurements (intraobserver reliability) was evaluated with Pearson correlation coefficients. The agreement of the measurements made by the 4 observers (interobserver reliability) and the agreement of the measurements of the same bone in different reconstruction planes (interplane reliability) were evaluated using interclass correlation coefficients. RESULTS: The intraobserver reliability was poor for 27 of 36 comparisons. The interobserver reliability of the dorsal cortical angle and the intrascaphoid angle was poor for all reconstruction planes. The interobserver reliability of the height-to-length ratio was good for 2 planes and poor for the third plane. The interplane reliability was poor for 7 of 12 comparisons, with no single measurement technique remaining consistent for all observers across reconstruction planes. CONCLUSIONS: Quantitative measurements of scaphoid deformity have very limited reliability for individual observers, between different observers, and depending on the plane in which the image of the scaphoid is produced. Even the most reliable measure of deformity (height-to length ratio) was not consistent between reconstruction planes. Unless more reliable scanning and measurement techniques are developed ideas about the effect of scaphoid deformity on wrist function will remain to a large degree speculative. PMID- 16039361 TI - Avulsion fracture of the hamulus from clay gunshot sport: a case report. AB - We report a case of hamulus (hook of hamate) fracture nonunion and secondary flexor digitorum profundus tendon rupture caused by repetitive wrist hyperextension sustained during clay shotgun shooting. The hyperextension caused avulsion of the hamulus by the pisohamate ligament. The hamulus was excised and a flexor digitorum profundus tendon transfer of the ring to small finger side to side was performed with satisfactory outcome. PMID- 16039362 TI - Symptomatic carpal collapse after trapeziectomy and partial trapeziodectomy: report of two cases. AB - We present 2 cases of symptomatic nondissociative dorsal intercalated segment instability (CIND-DISI) collapse after trapeziectomy and partial trapezoidectomy for pantrapezial arthritis. Preoperative radiographs showed normal carpal alignment in 1 case and a mild DISI alignment in the other. After surgery both developed increased CIND-DISI and carpal symptoms that differed from their original thumb-base problems. One required carpal realignment and fusion; to date the other has responded to nonsurgical treatment. Such problems may be recognized insufficiently by surgeons and thus may be more common than the current literature suggests. PMID- 16039363 TI - Dynamic effects of joint-leveling procedure on pressure at the distal radioulnar joint. AB - PURPOSE: Wrist joint-leveling procedures for decompression of the radiocarpal and ulnocarpal joints are accompanied by the risk for subsequent disorders of the adjacent distal radioulnar joint (DRUJ). This study evaluated the dynamic change of the pressure pattern at the DRUJ after joint-leveling procedures. METHODS: Thirteen fresh-frozen adult cadaveric upper extremities were used. A segment of the radius was excised at its midshaft to allow lengthening and shortening via a mini external fixator attachment. Dynamic pressure sensors were inserted into the DRUJ and the ulnocarpal joint. Axial loads were applied to the extensor carpi radialis brevis, extensor carpi radialis longus, extensor carpi ulnaris, flexor carpi radialis, and flexor carpi ulnaris for a total of 89 N with or without 30 N of radioulnar loading. The dynamic pressure distribution for full range of forearm rotation was recorded from 6 mm of radial shortening to 6 mm of radial lengthening in increments of 1 mm. RESULTS: The peak pressures at the DRUJ before the joint-leveling procedures averaged 3.3 MPa without radioulnar loading and 5.0 MPa with radioulnar loading. The peak pressures with axial and radioulnar loading increased 85% at 6 mm of lengthening and only 8% at 6 mm of shortening. The peak pressures at the DRUJ for radial lengthening of 4 mm or more were significantly greater than that of the original length. Pressure at the ulnocarpal joint increased in proportion to the amount of radial shortening and decreased with radial lengthening. CONCLUSIONS: Radial lengthening but not radial shortening significantly increases the peak pressure at the DRUJ. PMID- 16039364 TI - Ulnar shortening effect on distal radioulnar joint stability: a biomechanical study. AB - PURPOSE: The ulnar-shortening procedure has been adopted widely to reduce pressure between the ulna and ulnar carpus in ulnocarpal abutment syndrome. The hammock-like structure of the triangular fibrocartilage complex (TFCC), which supports and connects the ulnocarpal and distal radioulnar joint (DRUJ), variably is torn in this condition. The degree to which the torn TFCC may be tensioned to restabilize the DRUJ with ulnar recession is uncertain. This study examined changes in the stabilizing effect of the ulnar-shortening procedure in several TFCC tear conditions. METHODS: Six fresh-frozen cadaver arms amputated at the midportion of the humerus were used. The skin, muscles, and capsuloligamentous structures below the elbow all were preserved. The ulna and humerus were affixed firmly to a custom mount that allowed 60 degrees of forearm rotation. An external fixator was attached to the distal ulna leaving space for a 10-mm resection of the ulna to allow progressive shortening. The radius was attached to a materials testing machine. The load-displacement curves were obtained while translating the distal radius dorsally or palmarly with respect to the ulna at 1.25 mm/s. Stiffness in dorsopalmar displacement was recorded at 1-mm intervals through 6 mm of length. These measurements then were compared with controls (0 mm shortening of the intact specimens) at 60 degrees pronation, neutral position, and 60 degrees supination. The tests then were repeated after sectioning either the dorsal or palmar portion of the radioulnar ligament (RUL) and then after complete sectioning of the RUL. Each portion was sectioned at its attachment to the ulnar fovea. RESULTS: The stiffness of the DRUJ increased significantly in all 3 rotatory positions after shortening the ulna. A shortening of 6 mm resulted in a 26% to 44% increase in DRUJ stiffness. The stiffness decreased after partial sectioning of the RUL but increased with further ulnar shortening in all 3 positions. The DRUJ stiffness with the partially sectioned RUL after a shortening of 3 to 6 mm was as large as that of the intact specimens. The stiffness of the DRUJ after the complete section of the RUL was significantly smaller than that of the intact specimens even after shortening of 6 mm. CONCLUSIONS: The ulnar shortening procedure can stabilize the DRUJ by increasing intrastructural tension of the TFCC, only when the RUL is attached totally or partially to the ulnar fovea. If the RUL is avulsed completely then stability of the DRUJ no longer is obtained by the ulnar-shortening procedure. PMID- 16039365 TI - Reconstructive procedure for unstable radial-sided triangular fibrocartilage complex avulsions. AB - PURPOSE: Radial-sided avulsions of the triangular fibrocartilage complex (TFCC) (Palmer 1D) with distal radioulnar joint (DRUJ) instability remain a challenging pathology to treat. We tested an intra-articular reconstruction that addresses unstable radial-sided TFCC avulsions. METHODS: Ten preserved, dissected, cadaveric forearm specimens with intact TFCC and without ulnar-positive variance had biomechanical testing using a hydraulic testing device. The measurement of total displacement of the ulna relative to the radius was performed with an applied load ranging from 20 N in a volar direction to 20 N in a dorsal direction. Specimens were tested sequentially with intact TFCC, with surgically induced Palmer 1D lesions, and after reconstruction of the TFCC. All tests were performed at neutral, maximal pronation, and maximal supination. RESULTS: The mean total displacements of the DRUJ of the specimens at neutral rotation were as follows: 4.1 +/- 0.4 mm for the intact specimens compared with 11.8 +/- 0.8 mm after creation of the tear and 3.9 +/- 0.7 mm for the reconstructed specimens. In maximal pronation the mean total displacements were as follows: 2.4 +/- 0.3 mm intact versus 4.9 +/- 0.7 mm for torn and 2.1 +/- 0.3 mm after reconstruction. In maximal supination the mean total displacements were as follows: 1.4 +/- 0.2 mm intact versus 5.7 +/- 1.3 mm for torn and 1.0 +/- 0.1 mm after reconstruction. All specimens obtained the preoperative pronation and supination motion after the reconstruction. CONCLUSIONS: Current procedures are unable to restore DRUJ stability without a significant limitation of pronation and supination. This intra-articular reconstruction of radial-sided TFCC avulsions succeeded in restoring baseline stability to the DRUJ without interfering with pronation/supination. PMID- 16039366 TI - Comparison of distal radioulnar joint reconstructions using an active joint motion simulator. AB - PURPOSE: Distal radioulnar joint (DRUJ) instability can result in pain and functional disability. Numerous DRUJ reconstructive options have been described with minimal biomechanical analysis. The purpose of this study was to evaluate the ability of 4 well-described DRUJ reconstructions to restore joint kinematics using a dynamic, motion-controlled simulator. METHODS: Eleven cadaveric upper extremities had computer-controlled simulated active forearm rotation. Joint kinematics were quantified by using an electromagnetic tracking system. We compared the passive and simulated active kinematics of the intact, unstable, and reconstructed DRUJ (capsular repair, 2 described radioulnar ligament reconstructions, and a radioulnar tethering procedure). RESULTS: All reconstructions improved significantly the kinematics of the unstable DRUJ. The capsule repair restored simulated active joint kinematics closest to the intact DRUJ. CONCLUSIONS: All 4 reconstructions improved DRUJ stability significantly. The capsule repair most closely matched intact DRUJ kinematics and the radioulnar ligament reconstructions were found to be superior to a radioulnar tethering procedure. PMID- 16039367 TI - Volar fixation of dorsally displaced distal radius fractures using the 2.4-mm locking compression plates. AB - PURPOSE: To determine whether volar fixed-angle plate fixation with a new plate system could be used to treat dorsally unstable distal radius fractures. We hypothesized that volar fixed-angle plate fixation with or without radial styloid fixed-angle plate fixation would provide sufficient rigidity to allow early active range of motion without compromising fracture reduction. The initiation of early active motion may improve functional outcomes. METHODS: A retrospective review was conducted of one institution's initial experience using a new volar fixed-angle plate system to treat dorsally displaced intra-articular and extra articular distal radius fractures. Thirty-two fractures in 32 patients with dorsally displaced distal radius fractures were treated with a volar fixed-angle plate with or without a radial styloid fixed-angle plate. Fractures were classified using the AO classification. Radiographic parameters on preoperative, postoperative, and final follow-up radiographs were compared. The time to initiation of active range of motion was determined. Final follow-up ranges of motion and complications were reported. Finally, comparisons were made between the 23 fractures treated with a volar plate alone and the 9 fractures treated with a volar plate and a radial styloid plate. RESULTS: The average follow-up period was 13 months. Two thirds of the fractures were intra-articular. Average loss of reduction from initial postoperative to final follow-up radiographs was 0 degrees of volar tilt, 1 degrees of radial inclination, and 0 mm of radial length. Active wrist and forearm ranges of motion were initiated at an average of 11 days after surgery. The final follow-up flexion-extension and pronation supination arcs averaged 112 degrees and 151 degrees , respectively. The 9 fractures treated with the combination of a fixed-angle volar plate with a fixed angle radial styloid plate had greater initial displacement than did the 23 fractures treated with a volar plate alone. Otherwise, differences between the 2 groups were not significant. Only 1 radial styloid plate became symptomatic. CONCLUSIONS: Volar plate fixation using a new fixed-angle plate system successfully can stabilize dorsally unstable distal radius fractures. Early active range of motion was facilitated without compromising fracture reduction. PMID- 16039368 TI - Palmar plating system for Colles' fractures--a preliminary report. AB - PURPOSE: To present our results of the palmar plating system that we developed for the treatment of Colles' fractures. METHODS: By using the palmar plating system that we developed a consecutive series of 40 acute Colles' fractures were treated surgically. There were 12 men and 28 women with a mean age of 57 years at the time of the injury (range, 25-90 y). All patients had internal fixation using the trans-flexor carpi radialis tendon approach. The system has 3 main features. First, the plate is small in size, being 1.1 mm in thickness and 47 mm in length. Only a 3- to 4-cm skin incision is required for application of the plate. Second, the screw is cannulated and cancellous in type, with a low-profile head. Subchondral screw fixation is achieved both easily and safely by using a guidewire. Third, the plate has a window through which injectable bone cement can be placed. RESULTS: Union was achieved in all patients. The palmar tilt, radial inclination, radial length, and ulnar variance were maintained after surgery. According to the Gartland and Werley rating scale that was modified by Sarmiento there were 12 excellent and 28 good results. There were no extensor tendon injuries that could occur when the dorsal approach was used. CONCLUSIONS: This palmar plating system can make fixation of the distal radius easy, safe, and effective in the treatment of unstable Colles' fractures. PMID- 16039369 TI - Locking versus nonlocking T-plates for dorsal and volar fixation of dorsally comminuted distal radius fractures: a biomechanical study. AB - PURPOSE: To see if locking volar plates approach the strength of dorsal plates on a dorsally comminuted distal radius fracture model. Volar plates have been associated with fewer tendon complications than dorsal plates but are thought to have mechanical disadvantages in dorsally comminuted distal radius fractures. Locking plates may increase construct strength and stiffness. This study compares dorsal and volar locking and nonlocking plates in a dorsally comminuted distal radius fracture model. METHODS: Axial loading was used to test 14 pairs of embalmed radii after an osteotomy simulating dorsal comminution and plating in 1 of 4 configurations: a standard nonlocking 3.5-mm compression T-plate or a 3.5-mm locking compression T-plate applied either dorsally or volarly. Failure was defined as the point of initial load reduction caused by bone breakage or substantial plate bending. RESULTS: No significant differences in stiffness or failure strength were found between volar locked and nonlocked constructs. Although not significant, the stiffness of dorsal locked constructs was 51% greater than that of the nonlocked constructs. Locked or nonlocked dorsal constructs were more than 2 times stiffer than volar constructs. The failure strength of dorsal constructs was 53% higher than that of volar constructs. Failure for both volar locked and nonlocked constructs occurred by plate bending through the unfilled hole at the osteotomy site. Failure for both dorsal locked and nonlocked constructs occurred by bone breakage. CONCLUSIONS: Locking plates failed to increase the stiffness or strength of dorsally comminuted distal radius fractures compared with nonlocking plates. Failure strength and stiffness are greater for locked or nonlocked dorsal constructs than for either locked or nonlocked volar constructs. Whether the lower stiffness and failure strength are of clinical significance is unknown. The unfilled hole at the site of comminution or osteotomy is potentially a site of weakness in both volar locked and nonlocked plates. PMID- 16039370 TI - A randomized prospective study on the treatment of intra-articular distal radius fractures: open reduction and internal fixation with dorsal plating versus mini open reduction, percutaneous fixation, and external fixation. AB - PURPOSE: To compare 2 methods of surgical treatment for displaced intra-articular fractures of the distal radius: open reduction and internal fixation with dorsal plating (Pi Plate; Synthes, Paoli, PA) versus mini open reduction with percutaneous K-wire and external fixation. METHODS: Patients with AO type C intra articular distal radius fractures were randomized into 2 groups: open reduction and internal fixation and dorsal plating or external fixation and K-wires and mini-open reduction. Patients over the age of 70 years with any associated soft tissue or skeletal injury to the same limb and pre-existing wrist arthrosis or disability were excluded from the study. Objective, subjective, and radiographic outcomes were assessed at 2 weeks, 4 to 6 weeks, 10 to 12 weeks, 6 months, and 1- and 2-year intervals. The minimum follow-up period was 6 months; the average follow-up period was 18 months. The principal outcome analyzed was the Disabilities of the Arm, Shoulder, and Hand score. Secondary outcomes included grip strength, range of motion, surgical procedure time, complications, and radiographic parameters. The groups were equal with respect to age, gender, fracture subtype, and number of workers' compensation cases. RESULTS: No significant difference was found in the Disabilities of the Arm, Shoulder, and Hand scores, our primary outcome. The dorsal plate group, however, showed a higher complication rate when compared with the external fixator group. The plate group had significantly longer tourniquet times when compared with the external fixator group. The plate group also had higher levels of pain at 1 year when compared with the external fixator group; however, this equalized after hardware removal. The external fixator group showed an average grip strength of 97% compared with the normal side and 86% in the dorsal plate group. CONCLUSIONS: At midterm analysis the dorsal plate group showed a significantly higher complication rate compared with the external fixator group; therefore enrollment in the study was terminated. The dorsal plate group also showed statistically significant higher levels of pain, weaker grip strength, and longer surgical and tourniquet times. Based on these results we cannot recommend the use of dorsal plates in treating complex intra-articular fractures of the distal radius. PMID- 16039371 TI - The effect of increasing distraction on digital motion after external fixation of the wrist. AB - PURPOSE: Although several factors have been related to the development of digital stiffness after external fixation of distal radius fractures, distraction can be considered one of the most important. This study determined a threshold for distraction during external fixation of distal radius fractures and documented the effect of distraction of the wrist on metacarpophalangeal (MCP) joint motion. METHODS: Eight fresh-frozen cadaveric forearms were mounted on a testing frame and the flexor digitorum superficialis (FDS) tendon of each finger was pulled individually until a 90 degrees flexion at the MCP joint was generated. A load cell was used to record the force applied on the tendons. Tendon displacement was calculated by using a linear potentiometer. Sequential distraction across the wrist was applied through a distraction outrigger on the external fixator and was measured both on the external fixator in millimeters and on fluoroscopic images. Finger motion was captured by using a 2-dimensional motion tracking system. The experiment was repeated at different levels of wrist distraction (increments of 2 mm to a maximum of 14 mm). RESULTS: There was a significant difference in the effect of distraction on the individual fingers. For the index finger the load required for the FDS to produce a 90 degrees flexion at the MCP joint was increased linearly between 0 and 8 mm of wrist distraction. For the other fingers a threshold of distraction was found. A significant increase in the load was noticed after 5 mm of wrist distraction. CONCLUSIONS: More than 5 mm of wrist distraction increases the load required for the FDS to generate MCP joint flexion for the middle, ring, and small fingers. For the index finger, however, as much as 2 mm of wrist distraction significantly increases the load required for flexion at the MCP joint. PMID- 16039372 TI - Refined myoelectric control in below-elbow amputees using artificial neural networks and a data glove. AB - PURPOSE: To develop a system for refined motor control of artificial hands based on multiple electromyographic (EMG) recordings, allowing multiple patterns of hand movements. METHODS: Five subjects with traumatic below-elbow amputations and 1 subject with a congenital below- elbow failure of formation performed 10 imaginary movements with their phantom hand while surface electrodes recorded the EMG data. In a training phase a data glove with 18 degrees of freedom was used for positional recording of movements in the contralateral healthy hand. These movements were performed at the same time as the imaginary movements in the phantom hand. An artificial neural network (ANN) then could be trained to associate the specific EMG patterns recorded from the amputation stump with the analogous specific hand movements synchronously performed in the healthy hand. The ability of the ANN to predict the 10 imaginary movements offline, when they were reflected in a virtual computer hand, was assessed and calculated. RESULTS: After the ANN was trained the subjects were able to perform and control 10 hand movements in the virtual computer hand. The subjects showed a median performance of 5 types of movement with a high correlation with the movement pattern of the data glove. The subjects seemed to relearn to execute motor commands rapidly that had been learned before the accident, independent of how old the injury was. The subject with congenital below-elbow failure of formation was able to perform and control several hand movements in the computer hand that cannot be performed in a myoelectric prosthesis (eg, opposition of the thumb). CONCLUSIONS: With the combined use of an ANN and a data glove, acting in concert in a training phase, amputees rapidly can learn to execute several imaginary movements in a virtual computerized hand, this opens promising possibilities for motor control of future hand prostheses. PMID- 16039373 TI - Functional improvement with digital prosthesis use after multiple digit amputations. AB - PURPOSE: Patients who sustain traumatic amputation of multiple fingers suffer both a functional and psychologic loss. Previous studies of prosthesis use for finger amputees have focused primarily on the psychologic benefits. Clinically our group noticed a functional improvement on hand function tests when patients with multiple digit amputations used a prosthesis. Given the expense of multiple finger prostheses we sought to determine if they led to a consistent functional improvement in these patients. METHODS: Ten consecutive patients performed a battery of hand function tests and rated their ability to perform a variety of activities of daily living both with and without their prosthesis using the Disabilities of the Arm, Shoulder, and Hand questionnaire. RESULTS: Our results show a significant improvement in 3-finger-pinch strength and grip strength and a trend of improvement of tip-pinch, lateral-pinch, and grip strength in dynamometer positions 1, 2, 3, and 4 in these patients when tested with and without their prostheses. Function in activities of daily living, as assessed by the Disabilities of the Arm, Shoulder, and Hand questionnaire, was improved globally with prosthesis use. In addition, significant improvement was noted in several specific activities including opening a jar, writing, and turning a key, among others. CONCLUSIONS: These results show that prosthesis use provides a functional benefit to these patients in multiple activities. PMID- 16039374 TI - Radiation exposure to the hands from mini C-arm fluoroscopy. AB - PURPOSE: To quantify the level of radiation exposure to the hands of hand surgeons using intraoperative mini C-arm fluoroscopy and to compare the actual level of exposure with predicted levels and acceptable limits. METHODS: Five hand surgeons were given ring dosimeters to measure radiation exposure to their hands during surgery of the finger, hand, and wrist. A total of 81 rings were analyzed. After the clinical study a phantom was used to measure scatter at close range from the mini C-arm. RESULTS: Surgeons' hands were exposed to an average +/- SD of 20 +/- 12.3 mrem/case. For comparison a chest x-ray results in approximately 20 mrem exposure to the patient. Radiation exposure for the group of hand surgeons ranged from 5 to 80 mrem. Surgeons used an average of 51 +/- 36.9 seconds of fluoroscopy time per case. Exposure time for the group ranged from 6 to 170 seconds. The radiation scatter rate decreases precipitously outside the beam or beyond the radius of the intensifier. An average exposure to the hands of 20 mrem/case suggests that surgeons' hands must be entering the beam and getting direct exposure. CONCLUSIONS: Hand surgeons work close to the beam and as a result their hands potentially are exposed to a nontrivial amount of radiation. We recommend that surgeons who use the mini C-arm use precautions to minimize radiation exposure, particularly to their hands. PMID- 16039375 TI - Radiofrequency ablation in osteoid osteoma of the finger. AB - The occurrence of osteoid osteomas of the hand is rare and their treatment usually is surgical. A 26-year-old man with an osteoid osteoma in the proximal phalanx of the right middle finger was treated with percutaneous radiofrequency ablation. Two years later he remains free of pain and finger function is normal. PMID- 16039376 TI - Topographic localization of the motor branch of the median nerve. AB - PURPOSE: To redefine the localization of the thenar branch (TB) of the median nerve in relation to the surface landmarks that are in routine use. METHODS: The study was performed in 37 hands of 34 patients who had carpal tunnel release. All of the patients were women and the mean age was 50 years (range, 35-67 y). A radiologic marking technique was used to determine the localization of the TB, the middle finger radial side line, and Kaplan's cardinal line. The TB was marked by circumscribing with a soft radiopaque yarn and the surface landmark lines were shown by taping a K-wire to the hand for each line. An image-intensifier-printed image was obtained for each case and the distances between the markers of the TB and the wires were measured. RESULTS: The TB had a mean ulnar offset of 12.6 mm (range, 4.0-19.7 mm) from the middle finger radial side line and was located 4.4 mm (range, 0-9.5 mm) proximal to the cardinal line. CONCLUSIONS: During carpal tunnel release surgery the surgeon must pay more attention to the localization of the TB of the median nerve because it was found to be 12.6 mm more ulnar than the location described in the literature. PMID- 16039377 TI - Identification of the spinal accessory nerve within the surgical field during primary exploration of the brachial plexus in infants with obstetric palsy. AB - This report describes our technique for identifying the spinal accessory nerve during primary exploration of the brachial plexus in infants with obstetric palsy. PMID- 16039378 TI - Restoration of prehension using double free muscle technique after complete avulsion of brachial plexus in children: a report of three cases. AB - PURPOSE: Brachial plexus injury in children, excluding birth palsy, is relatively rare and seldom reported. We report our technique, the results of this procedure, and problems we encountered in treating children with brachial plexus injury. METHODS: From 1999 through 2002, we treated 3 children with complete avulsion of the brachial plexus due to trauma by using double free muscle technique (DFMT) with a nerve transfer procedure using the contralateral seventh cervical nerve root transfer to reconstruct prehensile function. There were 2 boys aged 5 and 11 years and a girl aged 4 years. All patients were followed up for at least 3 years after the surgery. RESULTS: All the transferred muscles survived without any vascular complications and were reinnervated successfully. The average active range of elbow flexion was 125 degrees (range, 90 degrees - 145 degrees ). The average total active range of motion of the fingers was 69 degrees (range, 40 degrees -102 degrees ). All patients obtained voluntary prehensile function and could use the reconstructed hand for activities of daily living. They were able to lift and carry light objects with the reconstructed hand and heavy objects with both hands. CONCLUSIONS: The results of DFMT for reconstruction of BPI in children were encouraging. Appropriate postoperative rehabilitation under close supervision is important to obtain useful prehensile function. PMID- 16039379 TI - Intraneural perineurioma of the radial nerve in a child. AB - An intraneural perineurioma is an uncommon solitary neoplasm of major nerve trunks composed of perineurial cells from the peripheral nerve sheath. The typical course of intraneural perineurioma is indolent, with gradual-onset motor loss and presentation for evaluation months to years after onset of symptoms. We report a 9-year-old girl with an intraneural perineurioma of the radial nerve. Diagnosis was confirmed by histology and immunohistochemistry. Resection and nerve graft reconstruction were performed to prevent progression of motor compromise and to allow recovery of motor function. An algorithm for the management of focal intraneural tumors is shown. PMID- 16039380 TI - Inhibition of inducible nitric oxide synthase promotes recovery of motor function in rats after sciatic nerve ischemia and reperfusion. AB - PURPOSE: To investigate the effects of inhibition of inducible nitric oxide synthase (iNOS) on the recovery of motor function in the rat sciatic nerve after ischemia and reperfusion injury. METHODS: A 10-mm segment of the sciatic nerve from 169 rats had 2 hours of ischemia followed by up to 42 days of reperfusion. The animals were divided into 2 groups that received either iNOS inhibitor 1400W or the same volume of sterile water subcutaneously. A walking track test was used to evaluate the motor functional recovery during reperfusion. Statistical analysis was performed for the measurements of the sciatic functional index (SFI) by using 2-way analysis of variance; 1-way analysis of variance was used for the post hoc analysis of specific values at each time point of the SFI measurement. RESULTS: 1400W-treated rats had earlier motor functional recovery than controls, with a significantly improved SFI between days 11 and 28. Histology showed less axonal degeneration and earlier regeneration of nerve fibers in the 1400W group than in the controls. Inducible NOS messenger RNA and protein were up-regulated during the first 3 days of reperfusion but there was a down-regulation of neuronal NOS and up-regulation of endothelial NOS in control animals. 1400W treatment attenuated the increase of iNOS but had no effect on neuronal NOS and endothelial NOS. CONCLUSIONS: Our results indicate that early inhibition of iNOS appears to be critical for reducing or preventing ischemia and reperfusion injury. PMID- 16039381 TI - A biomechanical analysis of suture materials and their influence on a four-strand flexor tendon repair. AB - PURPOSE: Flexor tendon repair strength depends on the suture technique and the suture material used. Configurations that incorporate locking loops prevent sutures from pulling through the tendon but typically fail because of suture breakage. The choice of suture material therefore influences repair strength. This study investigated the mechanical properties of 5 nonabsorbable 4-0 suture materials (monofilament nylon, monofilament polypropylene, braided polyester, braided stainless steel wire, and braided polyethylene) and evaluated their performance when used in a locking 4-strand flexor tendon repair configuration. METHODS: Five samples of 2 strands of each suture type were tested mechanically to determine the material stiffness and ultimate load. In addition, 50 fresh porcine flexor tendons were divided and repaired with each of the 5 suture materials using a 4-strand single-cross technique. Gap force, ultimate strength, and stiffness were measured to compare biomechanical performance. RESULTS: All repairs failed by suture rupture at the locking loop. Fibrewire and stainless steel sutures and repairs were significantly stronger and stiffer than the other suture types. The results for Prolene and Ethibond were similar in the tendon repair groups with respect to gap and ultimate forces although Ethibond provided significantly increased repair stiffness. Nylon sutures and repairs consistently produced the poorest mechanical performance in all outcome measures. CONCLUSIONS: Suture material strongly influences the biomechanical performance of multistrand tendon repairs and is an important consideration for the surgeon. Fibrewire and stainless steel are the most biomechanically suitable suture materials for flexor tendon repair whereas nylon is the least suitable. Further developments in suture materials are important for advancements in flexor tendon repair strength. PMID- 16039383 TI - Open traumatic avulsion of the flexor pollicis longus tendon from the musculotendinous area: a case report. AB - A traumatic avulsion of the flexor tendon at the musculotendinous junction in nonamputated digits is a very rare injury. We present a 14-year-old girl who sustained a longitudinal, tensile, injurious force directly to the flexor pollicis longus tendon after an open thenar injury resulting in its avulsion at the musculotendinous junction. In an effort to minimize soft-tissue damage and preserve the transverse ligament of the carpus the tendon was retrieved through a separate forearm incision. Direct repair was made by encapsulation of the tendon into the muscle belly. The functional result 30 months after surgery was satisfactory. PMID- 16039382 TI - Clinical and radiographic evaluation of surgical reconstruction of finger flexion in tetraplegia. AB - PURPOSE: To define the order and frequency of elongation in tendon junctions in extensor carpi radialis longus (ECRL) to flexor digitorum profundus tendon transfer and its correlation to grip strength and lack of finger flexion. METHODS: Forty-seven tetraplegic patients had surgery involving the reconstruction of finger flexion with the transfer of the ECRL in a total of 62 arms. During surgery metal markers were placed on both sides of the tendon-to tendon attachment site. After surgery the distance between the markers was measured on radiographs. Any increase in the distance between the markers was judged as elongation. The grip strength and mean pulp-to-palm distance were evaluated a minimum of 6 months after surgery. Three arms had a second surgery because of insufficient functional results. RESULTS: The average final elongation was 9 +/- 10 mm (mean +/- SD). The mean grip strength was 16 +/- 12 kPa (range, 0 50). The lack of flexion (mean value of pulp-to-distal palmar crease of 3 ulnar digits) was 0.8 +/- 1.2 cm on average. Elongation up to 15 to 20 mm still was compatible with good grasp. CONCLUSIONS: The tendon junction after a transfer of the ECRL to the finger flexors can be overloaded. Elongation therefore must be considered as one among several possible causes of an unsatisfactory result after this type of tendon transfer but elongation less than 15 mm usually is compatible with excellent function. PMID- 16039384 TI - Osseous sarcoidosis of the hand: pathologic analysis and review of the literature. AB - A destructive granulomatous process involving the right fifth metacarpal and the soft tissues of the right thumb and nose of an African-American woman without pulmonary disease is described. The initial biopsy examination of the metacarpal showed caseating and noncaseating granulomata. After a fifth-ray amputation the disease progressed, leading to the referral of the patient to our institution. A biopsy examination of this recurrence showed a predominance of solid noncaseating granulomata. The diagnosis of sarcoidosis was made on the basis of the morphology of the granulomata and by exclusion of infectious and neoplastic causes. Steroid therapy has resulted in cessation of clinical and radiographic disease progression at a 3-year clinical follow-up evaluation. PMID- 16039385 TI - Use of the anterolateral thigh free flap for upper-extremity reconstruction. AB - PURPOSE: The anterolateral thigh free flap (ALTF) first was reported in 1984 and has been used in large series with success for a multitude of clinical purposes. We describe our results with the ALTF in upper-extremity and hand reconstruction. METHODS: From 1996 to 2003 there were 15 patients who had reconstruction of the hand and upper extremity using the ALTF. The parameters used to assess the outcome of our series included the success rate of the flap as measured by flap survival rate and adequacy of skin coverage, ability to close the donor site primarily or necessity of a skin graft, complications associated with the flap, donor site, and non-flap-related complications such as pulmonary embolism. RESULTS: Of the 15 patients with an ALTF, 3 (20%) had a musculocutaneous perforator and 12 (80%) had a septocutaneous perforator. Two patients had a neurotized ALTF reconstruction. There were 4 complications related to the flap with 1 complete flap failure. The overall flap survival rate was 93%. The donor site was closed primarily in 8 patients (53%) and with a skin graft in 7 patients (47%). One donor site breakdown occurred. CONCLUSIONS: Our results show many advantages of the ALTF for upper-extremity reconstruction. Because of its versatility the ALTF is suited ideally for upper-extremity reconstruction and should be considered as part of the reconstructive ladder. PMID- 16039386 TI - FCU anatomy. PMID- 16039387 TI - Southern California Journal Club Review award. PMID- 16039388 TI - Carpal tunnel syndrome in patients more than 65 years old. PMID- 16039391 TI - Condone or condemn? PMID- 16039392 TI - Carpal Tunnel Syndrome and TMJ Arthritis. PMID- 16039394 TI - Stability of the distal radioulnar joint contributed by the joint capsule. PMID- 16039397 TI - The role of trivalent dimethylated arsenic in dimethylarsinic acid-promoted skin and lung tumorigenesis in mice: tumor-promoting action through the induction of oxidative stress. AB - We investigated the relationship between lung- and skin-tumor promotion and oxidative stress caused by administration of dimethylarsinic acid (DMA(V)) in mice. The incidence of lung tumors induced by lung tumor initiator (4NQO) and DMA(V) were, as well as 8-oxo-2'-deoxyguanosine (8-oxodG), suppressed by cotreatment with (-)epigallocatechin gallate (EGCG). When mice were topically treated with trivalent dimethylated arsenic (DMA(III)), a further reductive metabolite of DMA(V), not only an increase in skin tumors but also an elevation of 8-oxodG in epidermis were observed. These results suggest that tumor promotion due to DMA(V) administration is mediated by DMA(III) through the induction of oxidative stress. PMID- 16039398 TI - Gene expression changes in human small airway epithelial cells exposed to Delta9 tetrahydrocannabinol. AB - Marijuana smoking is associated with inflammation, cellular atypia, and molecular dysregulation of the tracheobronchial epithelium. While marijuana smoke shares many components in common with tobacco, it also contains a high concentration of Delta9-tetrahydrocannabinol (THC). The potential contribution of THC to airway injury was assessed by exposing primary cultures of human small airway epithelial (SAE) cells to THC (0.1-10.0 microg/ml) for either 1 day or 7 days. THC induced a time- and concentration-dependent decrease in cell viability, ATP level, and mitochondrial membrane potential. Using a targeted gene expression array, we observed acute changes (24 h) in the expression of mRNA for caspase-8, catalase, Bax, early growth response-1, cytochrome P4501A1 (CYP1A1), metallothionein 1A, PLAB, and heat shock factor 1 (HSF1). After 7 days of exposure, decrease in expression of mRNA for heat shock proteins (HSPs) and the pro-apoptotic protein Bax was observed, while expression of GADD45A, IL-1A, CYP1A1, and PTGS-2 increased significantly. These findings suggest a contribution of THC to DNA damage, inflammation, and alterations in apoptosis. Treatment with selected prototypical toxicants, 2,3,7,8-tetrachlorodibenznzo-p-dioxin (TCDD) and carbonyl cyanide-p-(trifluoramethoxy)-phenyl hydrazone (FCCP), produced partially overlapping gene expression profiles suggesting some similarity in mechanism of action with THC. THC, delivered as a component of marijuana smoke, may induce a profile of gene expression that contributes to the pulmonary pathology associated with marijuana use. PMID- 16039399 TI - Metallothionein-enriched hepatocytes are resistant to ferric nitriloacetate toxicity during conditions of glutathione depletion. AB - Metallothionein (MT) is involved not only in heavy metal homeostasis/detoxification but also in radical scavenging, yet the relevance to other antioxidant systems and physiological significance under oxidative stress has not been clarified. We studied that ability of MT, induced by zinc and cadmium, to protect against oxidative damage induced by ferric nitrilotriacetate (Fe-NTA) in glutathione depleted primary cell cultures. Treatment with Fe-NTA resulted in significant decreases in cell survival and increases in medium LDH activity in control cells following depletion of glutathione. The toxic effects of Fe-NTA were modulated in Zn-MT-enriched cells. In glutathione-depleted cells, but not in non-treated cells, Cd-binding properties of cellular Zn-MT decreased with increasing concentration of Fe-NTA. Both Zn-MT and Cd-MT-enriched cells were resistant to higher doses of Fe-NTA. These results indicate that MT may act a cellular radical scavenger in the absence of GSH. Thus, MT may function as a secondary antioxidant in a cellular protection system. PMID- 16039400 TI - Vinyl acetate induces intracellular acidification in mouse oral buccal epithelial cells. AB - Vinyl acetate exposure in drinking water has been associated with tumor formation in the upper gastrointestinal tract of rats and mice. One potential mechanism for inducing carcinogenesis involves acidification of the intracellular environment due to the metabolism of vinyl acetate to acetic acid. Prolonged intracellular acidification is thought to produce cytotoxic and/or mitogenic responses that are the sentinel pharmacodynamic steps toward cancer. To determine whether exposure to vinyl acetate affects the intracellular pH of intact oral cavity tissue, isolated mouse oral buccal epithelium was loaded with the pH-sensitive dye BCECF, and then exposed to vinyl acetate concentrations ranging from 10 to 1000 microM for up to 4 min. Extracellular vinyl acetate exposure induced a progressive intracellular acidification that was reversible upon removal of the vinyl acetate. The rate of the acidification was concentration-dependent and increased exponentially within the concentration range tested. The magnitude of the vinyl acetate-induced acidification was inhibited by pretreatment with the carboxylesterase inhibitor bis(p-nitrophenyl)phosphate. These results are consistent with the hypothesis that vinyl acetate contributes to the generation and progression of oral cavity tumors via a process of intracellular acidification. Such a process has been proposed to have practical dose-response thresholds below which the intracellular environment can be maintained within homeostatic bounds and the contribution of exposure to carcinogenic risk is negligible. PMID- 16039401 TI - Particle surface characteristics may play an important role in phytotoxicity of alumina nanoparticles. AB - The phytotoxicity of alumina nanoparticles loaded with and without phenanthrene (Phen) was investigated by means of root elongation (RE) experiments in this study. Five plant species, Zea mays (corn), Cucumis sativus (cucumber), Glycine max (soybean), Brassica oleracea (cabbage), and Daucus carota (carrot) were used in our study of phytotoxicity by root elongation experiments. The surface characteristics of Phen-loaded and Phen-nonloaded nanoparticles were investigated using the Fourier transformed infrared (FTIR) spectroscopy technique. It was found that when loaded with 10.0%, 100.0%, or 432.4% monomolecular layer (MML) of Phen, the degree of the root elongation inhibition caused by the particles was reduced. The loading of Phen leads to the appearance of a vibrational mode in the region of 850-1050 cm(-1), which was assigned to the surface characteristics of the particles and arises from the disappearance of free hydroxyl groups according to an earlier study. When mixed with a known free hydroxyl radical scavenger, DMSO (0.5% and 1.0%), the non-loaded particles also showed decreased inhibition of root elongations. We supposed that the surface characteristics of the particles play an important role in the phytotoxicity of alumina nanoparticles. PMID- 16039402 TI - The neurotoxic lipopeptide kalkitoxin interacts with voltage-sensitive sodium channels in cerebellar granule neurons. AB - The marine neurotoxin kalkitoxin, a thiazoline-containing lipid derived from the pantropical marine cyanobacterium Lyngbya majuscula, was assayed for interaction with the tetrodotoxin-sensitive, voltage-sensitive sodium channel (TTX-VSSC) in cerebellar granule neuron cultures (CGN). The naturally occurring isomer of kalkitoxin (KTx-7) blocked veratridine-induced (30 microM) neurotoxicity in a concentration-dependent manner (EC50 22.7 nM [9.5-53.9 nM, 95% confidence interval {CI}]) in CGN. Kalkitoxin was a potent inhibitor (EC50 26.1 nM [12.3 55.0 nM, 95% CI]) of the elevation of intracellular Ca2+ concentration [Ca2+](i) that accompanies exposure of CGN to veratridine. To further explore the potential interaction of KTx-7 with TTX-VSSC, we assessed the influence of KTX-7 on the binding of [3H]batrachotoxin ([3H]BTX) to neurotoxin site 2 on the TTX-VSSC. Although kalkitoxin was without effect on the basal binding of [3H]BTX to intact cerebellar granule neurons, in the presence of the positive allosteric modulator, deltamethrin, [3H]BTX binding was inhibited by KTx-7 in a concentration-dependent manner (11.9 nM [IC50=3.8-37.2 nM, 95% CI]). These results provide both direct and functional evidence for an interaction of kalkitoxin with the neuronal TTX VSSC. PMID- 16039403 TI - Soluble transition metals in welding fumes cause inflammation via activation of NF-kappaB and AP-1. AB - We previously reported that the molecular pro-inflammatory effects of welding fumes in vitro were caused by soluble transition metals via an oxidative stress mediated mechanism. Herein, we tested the hypothesis that transition metals in welding fume drive the in vivo inflammatory response caused by welding fume. Rats were instilled with either whole, soluble extract or washed welding fume particulates or soluble extracts pre-treated with a transition metal chelator. Markers of pulmonary inflammation were measured in the bronchoalveolar lavage fluid (BALF) and nuclear translocation of transcription factor was assessed in BAL cells by electrophoretic mobility shift assay. Instillation of either whole or soluble fractions of welding fume caused a significant influx of inflammatory cells and other markers of inflammation in the BALF 24 h later. MIP-2 protein in BALF and nuclear translocation of NF-kappaB and AP-1 were significantly greater following instillation of whole and soluble fractions than in saline-instilled lungs. Chelation of the soluble fraction, to remove transition metals, abolished the ability to cause inflammation, MIP-2 increase or transcription factor translocation to the nucleus. Instillation of washed particulates alone caused no significant change in any end-point compared to saline. This study demonstrates that soluble transition metals present in welding fumes cause inflammation via activation of the redox-sensitive transcription factors NF-kappaB and AP-1 and confirms the validity of utilising in vitro models to assess inflammatory responses to such particles. PMID- 16039404 TI - Endometrial cancer--surprising reports. PMID- 16039405 TI - Influence of hormone replacement therapy on proteomic pattern in serum of postmenopausal women. AB - OBJECTIVES: Proteomics approaches to cardiovascular biology and disease hold the promise of identifying specific proteins and peptides or modification thereof to assist in the identification of novel biomarkers. METHOD: By using surface enhanced laser desorption and ionization time of flight mass spectroscopy (SELDI TOF-MS) serum peptide and protein patterns were detected enabling to discriminate between postmenopausal women with and without hormone replacement therapy (HRT). RESULTS: Serum of 13 HRT and 27 control subjects was analyzed and 42 peptides and proteins could be tentatively identified based on their molecular weight and binding characteristics on the chip surface. By using decision tree-based Biomarker Patternstrade mark Software classification and regression analysis a discriminatory function was developed allowing to distinguish between HRT women and controls correctly and, thus, yielding a sensitivity of 100% and a specificity of 100%. The results show that peptide and protein patterns have the potential to deliver novel biomarkers as well as pinpointing targets for improved treatment. The biomarkers obtained represent a promising tool to discriminate between HRT users and non-users. CONCLUSION: According to a tentative identification of the markers by their molecular weight and binding characteristics, most of them appear to be part of the inflammation induced acute phase response. PMID- 16039406 TI - Differences between the pre-menopausal and post-menopausal uterine fibroid vasculature. AB - OBJECTIVES: To quantitatively examine differences in microvascular density between fibroid and myometrial tissue from fibroid uteri removed at hysterectomy, both before and after the menopause, and with hormone replacement therapy. METHODS: Factor VIII immunostaining of formalin fixed tissues was used to identify blood vessels, and the vessels counted by an investigator blinded to tissue type or menopausal status. RESULTS: The mean myometrial: fibroid MVD ratio was 2.38 higher in the post-menopausal group (95% CI: 0.12, 4.65, p=0.0474) than in the pre-menopausal group, with the hormone therapy (HT)-using post-menopausal group lying in between. An increase in microvascular density in the myometrium after the menopause was responsible for most of the change in ratios seen between the pre and post-menopausal pairs. There was a trend to increasing myometrial MVD with increasing number of years post-menopause. CONCLUSIONS: Myometrial microvascular density increases markedly after the menopause, while fibroid microvascular density does not alter. Thus, the difference between myometrial and fibroid vasculature becomes greater after the menopause. The implications of this for the treatment of fibroids in post-menopausal women is discussed. PMID- 16039407 TI - Prophylactic oophorectomy at elective hysterectomy. Effects on psychological well being at 1-year follow-up and its correlations to sexuality. AB - OBJECTIVE: In a prospective study, the 1-year psychological well-being outcome after oophorectomy-hysterectomy, compared to hysterectomy-only and the correlations between the changes in psychological well-being and the changes in sexuality were evaluated. STUDY POPULATION: Perimenopausal sexually active women (n=362), scheduled for hysterectomy on benign indication, were recruited. Three hundred and twenty-three women (89%) completed the 1-year follow-up; 217 women spared their ovaries and 106 underwent concomitant oophorectomy. METHODS: Psychological well-being was studied by the psychological general well-being index (PGWB) and sexuality by the McCoy's sex questionnaire. The prevalence of climacteric symptoms was reported by the modified Kupperman's index. Hormone replacement therapy was recorded. Postoperative, all oophorectomized and the hysterectomy-only women with climacteric symptoms were recommended estrogen replacement therapy. RESULTS: The two groups did not differ in PGWB, neither before surgery nor at 1-year follow-up. Postoperative, both groups showed increased well-being regarding depressed mood, general health and total score. Besides, the hysterectomy-only group had increased vitality and the hysterectomy oophorectomy group showed increased positive well-being and decreased anxiety. Most of the sexual parameters showed positive correlation to the PGWB parameters. The correlations were strong regarding parameters of overall sexual satisfaction, weak, regarding sexual motivation and relationship to partner, while absent regarding coital frequency. CONCLUSION: Concomitant prophylactic oophorectomy, at elective hysterectomy, does not negatively affect psychological well-being in adequately estrogenized perimenopausal women. Indeed, both hysterectomy-only and hysterectomy-oophorectomy have a positive effect on psychological well-being. Most aspects of sexuality are correlated to aspects of psychological well-being. PMID- 16039408 TI - Development of nocturia in relation to health, age and the menopause. AB - OBJECTIVES: To analyse the relationship between nocturia, age, the menopausal transition, parturition and hormone replacement treatment (HRT) in women. MATERIAL AND METHODS: A questionnaire study was carried out in 3669 randomly selected women (out of 6000 invited) in the County of Jamtland, Sweden. Questions were asked about health, the occurrence of somatic diseases and symptoms, habits, health care and medication. RESULTS: In univariate analyses age, menopausal state and HRT, but not parturition, were associated with increased nocturia. In a multiple logistic regression analysis, independent correlates for two or more nocturnal micturition episodes versus no more than one episode were: health, poor versus good 2.9 (2.1-3.8); <5 years after the menopause versus before 1.8 (1.3 2.5); 5-9 years after the menopause versus before 2.1 (1.5-3.0); > or =10 years after the menopause versus before 3.1 (2.3-4.2). Age and HRT were deleted by the logistic regression model. CONCLUSION: The present data indicate that menopausal state, but not age, parturition and HRT are independent correlates of nocturia in 40-64-year-old women. PMID- 16039409 TI - Effects of tibolone on body composition in postmenopausal women: a 1-year follow up study. AB - OBJECTIVE: We investigated the effects of 1-year tibolone treatment on body weight, body composition and indices of android obesity in postmenopausal women. METHODS: Forty-four postmenopausal women participated in this open-label controlled study; mean age was 51.8+/-2.21 years and all women were menopausal for 3.8+/-1.40 years. Twenty-two of them started taking 2.5 mg tibolone (TIB) daily for 1 year, whereas the remaining 22 served as age-matched controls. All subjects underwent a structured interview, physical examination, body composition measurements performed by dual-energy X-ray absorptiometry (DXA) - Hologic QDR 4500 A, as well as bioelectrical body impedance analysis (BI) - Tanita TBF-215, Japan. RESULTS: The TIB group did not significantly increase their weight (+0.4 kg), while the non-treated controls increased their mean weight by 1.4 kg (p=0.046). In the TIB group, DXA showed a non-significant body fat decrease by a mean of 0.5 kg and a non-significant lean mass increase by 0.8 kg, while in the control group, fat mass increased by 1.7 kg (p=0.032) and lean mass did not change. BI revealed that the TIB group had lost some fat ( approximately 0.6 kg, n.s.) and put some free-fat mass ( approximately 1.0 kg, p=0.048) without changes in total body water. The control group put on some fat ( approximately 1.1 kg, p=0.042) and lost some body water ( approximately 0.4 kg, n.s.). CONCLUSION: Results from both methods of measuring body composition show a similar trend: a decrease in fat mass and an increase in lean mass in TIB treated subjects. From the body composition perspective, tibolone may be regarded as a preferential alternative to conventional hormonal therapy (HT) in postmenopausal women. PMID- 16039410 TI - Morphometric evaluation of effects of two sex steroids on mammary gland of female rats. AB - OBJECTIVE: The purpose of this study was to evaluate the effects of two sex hormones on normal mammary gland of female rats. METHODS: Forty 250-day-old female rats, 20 of them with offspring and 20 not, were ovariectomized and, divided into 4 subgroups in order to receive one of the following subcutaneous treatment: estradiol benzoate (EB), medroxyprogesterone (MPA), EB+MPA or placebo, for 10 weeks. After treatment, mammary glands were studied with optical microscope. Whole gland, lobule, ductule and lumen compartments were evaluated by morphometric methods. Also a qualitative evaluation were performed seeking for secretion, microcalcification and trophic status. RESULTS: It was found that (a) MPA-only and placebo were similar for all parameters; (b) the same between EB and EB+MPA; (c) EB and EB+MPA increased lobule, ductule and lumen compartments significantly compared to MPA-only or placebo; (d) EB increased epithelium but without significance and EB+MPA increased it significantly compared to placebo or MPA; (e) EB and EB+MPA incremented secretion. CONCLUSIONS: In normal mammary gland of female rats: progestin action depends on estrogen presence. MPA does not revert estrogen-dependent proliferation, but it magnifies estradiol effect. Both EB and EB+MPA stimulate differentiation. Rats without offspring presented a greater epithelial proliferation under treatment with these sex hormones. PMID- 16039411 TI - Menopausal transition in Movima women, a Bolivian Native-American. AB - BACKGROUND: Although it is well accepted that socio-cultural influences may affect the onset of menopause and the symptoms experienced, there are scant data related to native indigenous populations. OBJECTIVE: Our purpose was to study in a native population of South America (The Movima, Bolivia) the menopausal transition. SUBJECTS AND METHODS: In this cross-sectional survey a total of 125 Movima women between 35 and 54 years old was included. All of them answered a questionnaire and a face-to-face interview focused on the menopausal experiences of middle-aged Movima women and additionally, history and physical examination, demographic and socio-economic information were obtained. RESULTS: The onset of natural menopause in the Movima was at 42.3+/-6.17 years. Loss of libido was the main complaint of the menopause, occurring in a 51% of interviewed women, hot flushes in a 45%, genital itching in a 40.8% and dyspareunia in a 40% were also frequently reported. CONCLUSION: This is the first data recorded on menopausal transition of the Movima natives. The age of menopause and the symptoms experienced are different to those from women of developed countries. Data related to minority groups are important to adapt the health system to their particular requirements. PMID- 16039412 TI - Skeletal status assessed by quantitative ultrasound at the calcaneus in females with bronchial asthma on prolonged corticosteroid therapy. AB - OBJECTIVE: The aim of the study was to assess skeletal status in bronchial asthma female patients after long-term oral corticosteroid (CS) therapy. METHODS: Eighty two female patients (25 with and 57 without fractures; mean age 58.0+/-7.8 years) were compared with 999 females (821 controls without fractures, mean age 58.6+/ 7.2 years and 178 females with previous osteoporotic fractures, mean age 57.8+/ 7.1 years). Mean age and years since menopause did not differ between patients and controls. The duration of CS therapy was 8.4+/-7.3 years, and daily mean dose equivalent to prednison was 8.8+/-3.5 mg. Skeletal status was evaluated by quantitative ultrasound (QUS) measurements at the heel using the Achilles system (Lunar, USA) which measures speed of sound (SOS (m/s)) and Broadband Ultrasound Attenuation (BUA (dB/MHz)). The Achilles software also calculates a stiffness index (SI (%)). The precision expressed using the root mean square coefficient of variation (RMS_CV%) was: 0.26% for SOS, 4.37% for BUA, and 2.13% for SI. RESULTS: Patients (all, with and without fractures) had significantly lower QUS values than controls without fractures and their values did not differ significantly from controls with fractures. Controls with fractures had significantly lower QUS values than controls without fractures. There was no difference between patients with and without fractures. The duration of the therapy did not influence skeletal variables in any group. Receiver operating characteristic curve (ROC) analysis was performed to assess the discriminatory capability of calcaneal QUS for CS-treated patients by calculating the area under the ROC curve (AUC). AUCs were: 0.70 for SOS, 0.68 for BUA and 0.70 for SI. CONCLUSIONS: In females with bronchial asthma on prolonged CS therapy, skeletal status is affected but does not differ from controls with fractures therefore CS therapy seems to be a risk factor for osteoporotic fracture. Calcaneal QUS measurements can be a useful tool in the assessment of CS bone-side effects. PMID- 16039413 TI - Seasonal onset of the menopause. AB - OBJECTIVES: A seasonal rhythm of reproduction is evident in humans. Herein it was investigated whether also the cessation of woman's fertile life follows a seasonal rhythm. METHODS: A retrospective study was performed on 2436 women in postmenopause for more than 12 months, in our menopause centres. Time of menopause was stratified for month and season. The variation was compared to the seasonal rhythm of 14,310 conceptions. RESULTS: The onset of menopause was more frequent (p<0.0001) in winter (32.5%) than in spring (20.8%), autumn (20.3%) and summer (26.2%), in which a minor peak was also observed (p<0.0001 vs. spring and autumn). The two peaks were temporally coincident with the transitions between the high to low and low to high rate of conceptions. CONCLUSIONS: The present data show that in women, like reproduction also the onset of menopause shows a seasonal modulation. PMID- 16039414 TI - Cimicifuga racemosa dried ethanolic extract in menopausal disorders: a double blind placebo-controlled clinical trial. AB - OBJECTIVES: To compare the efficacy and safety of the black cohosh root extract Cr 99 with placebo in women with climacteric complaints. METHODS: A multicenter, randomized, placebo-controlled, double-blind, parallel group study was conducted in 122 menopausal women (intention-to-treat population) with > or =3 hot flashes a day, treated over 12 weeks. Two main efficacy measures - weekly weighted score of hot flashes and Kupperman Index - and secondary efficacy variables, e.g. Menopause Rating Scale, were defined. Routine safety laboratory parameters and adverse events were documented. RESULTS: The primary efficacy analysis showed no superiority of the tested black cohosh extract compared to placebo. However, in the subgroup of patients with a Kupperman Index> or =20 a significant superiority regarding this index could be demonstrated (P<0.018). A decrease of 47% and 21% was observed in the black cohosh and placebo group, respectively. The weekly weighted scores of hot flashes (P<0.052) and the Menopause Rating Scale (P<0.009) showed similar results. Prevalence and intensity of the adverse events did not differ in the two treatment groups. CONCLUSIONS: The results indicate a superiority of the tested Cimicifuga racemosa extract compared to placebo in patients with menopausal disorders of at least moderate intensity according to a Kupperman Index > or =20, but not in the intention-to-treat population as a whole. PMID- 16039415 TI - The pups' endometrium morphology is affected by maternal malnutrition during suckling. AB - OBJECTIVES: This study aims to determine the effects of maternal protein and energy malnutrition during lactation on the endometrial structures of the offspring at puberty. METHODS: At parturition, dams were randomly assigned to the following groups: control group (C), with free access to a standard laboratory diet containing 23% protein; protein-restricted (PR) group, with free access to an isoenergy and protein-restricted diet containing 8% protein; and energy restricted (ER) group, receiving standard laboratory diet in restricted quantities. After weaning, all female pups had free access to standard laboratory diet. At puberty, the animals were sacrificed with pentobarbital and only females on the diestrum stage were used for the analyses. The stereological method used for quantifying the uterine endometrium was the M42 test system. RESULTS: When compared to C group, both PR and ER groups presented a significant reduction in the length density of the glands (PR=53%, ER=35.7%, p<0.001), in the volumetric density of the epithelium (PR=49%, ER=38%, p<0.001) and lumen (PR=42.7%, p<0.001; ER=23.8%, p<0.001) and in the surface density of the inner (PR=22%, ER=13.8%, p<0.001) and outer (PR=55.4%, p<0.01; ER=40.6%, p<0.001) glands. The volumetric density of the stroma was significantly higher in both PR (114%, p<0.001) and ER (117%, p<0.001) groups. In all parameters studied, there was no significant difference between PR and ER groups. CONCLUSIONS: Our results show that the protein and energy restriction during lactation leads to an atrophy of the uterine endometrial glands of the offspring at puberty. PMID- 16039416 TI - Women's views of the climacteric at the time of low menopausal hormone use, Estonia 1998. AB - OBJECTIVES: This study examined women's opinions about the climacteric and hormone therapy (HT) after menopause and compared women's and physicians' opinions in a country of low-HT use. METHODS: In 1998, a postal questionnaire was sent to a random sample of 2000 Estonian 45-64-year-old women; 69% (n=1312) responded. In 1999, a postal questionnaire was sent to a random sample of 500 Estonian gynaecologists and general practitioners; 68% (n=342) responded. RESULTS: Mean age at menopause was 49.8 years (S.D. 4.0), and there was no difference by socioeconomic classes or by age in self-rated health. Ten percent of women reported having used HT, with 3% currently using it. Most women reported some symptoms, with vasomotor symptoms more frequently reported by 50-54 years old; women most often reported tiredness (48%). Half of the women but under a fifth of physicians considered the climacteric a normal phase of life. Women's awareness about HT was low and about half had no opinion on its health effects. Half of the women had visited a gynaecologist, older women less so. Women with contacts with health care were more aware of HT. CONCLUSIONS: Women reported symptoms by age-group as similarly found in high-HT use countries and it verifies that many symptoms experienced were not due to menopause. As in other low-HT use countries, women were unfamiliar with HT and their attitudes were traditional, although physicians' attitudes were more positive. Estonian women seemed to have escaped the period of the preventive use of HT. PMID- 16039417 TI - Oxytocin decreases corticosterone and nociception and increases motor activity in OVX rats. AB - OBJECTIVES: In the present study the effects of oxytocin administered subcutaneously (s.c.) or intravaginally (i.vag.) on spontaneous motor activity, nociceptive thresholds and plasma corticosterone levels were examined in female ovariectomized (OVX) rats. METHODS: Oxytocin (1 mg/kg s.c. or 100 microg i.vag.) was administered once a day for 10 days to OVX rats. Controls received saline s.c. or cellulose gel i.vag. Spontaneous motor activity was observed in an open field arena, nociceptive thresholds were investigated by the tail-flick test, and corticosterone and oxytocin plasma levels were measured by radioimmunassay, 3, 4 and 5 days respectively, after the end of the treatment period. RESULTS: Both oxytocin administered s.c. and i.vag. increased forward locomotion (p<0.05) and nociceptive thresholds (p<0.05) significantly. In addition, oxytocin s.c. increased the amount of locomotor activity (p<0.05). Plasma corticosterone levels were decreased (p<0.05) and oxytocin levels were unchanged when measured 5 days after the last administration of oxytocin s.c. or i.vag. CONCLUSION: The present data indicate that oxytocin induces a spectrum of long-lasting effects in OVX rats, including an increase in spontaneous motor activity, elevation of nociceptive thresholds and decrease of corticosterone levels. Similar effects may be induced by estrogens. In addition, these data indicate that i.vag. administration of oxytocin may be used to induce oxytocin-mediated effects. PMID- 16039418 TI - Effect of OS-0689, a novel SERM, on periarterial nerve function in tail arteries of ovariectomized rats. AB - OBJECTIVE: We have previously shown that OS-0689 attenuates the rise in tail skin temperature of ovariectomized rats, which is believed to be relevant to human symptoms of hot flush. In this study, we elucidate the mechanism underlying the ameliorating effects of OS-0689 on elevated tail skin temperature. METHODS: Female Sprague-Dawley rats were ovariectomized and orally treated with OS-0544 (1 mg/kg), OS-0689 (3 mg/kg; (+)-enantiomer of OS-0544) or 17beta-estradiol (3 mg/kg; E2) for 1 week. At 1, 3 or 6 weeks after ovariectomy, the vasoconstrictions and vasorelaxations induced by periarterial nerve stimulation (PNS), l-noradrenaline (NA), and rat calcitonin gene-related peptide (CGRP) in isolated tail arteries were compared between OVX and sham-operated rats. RESULTS: Three weeks after ovariectomy, vasoconstrictions in response to PNS and NA in the arteries of OVX rats were markedly less than those in the arteries of sham operated rats. However, at 1 and 6 weeks after ovariectomy the stimuli-induced vasoconstrictions in the arteries of OVX rats were greater than those of sham operated rats. Moreover, NA reactivity was not attenuated in the mesenteric arteries at 3 weeks after ovariectomy. OS-0544, OS-0689 and E2 prevented the decrease in vasoconstrictions in the tail arteries. Vasorelaxations in response to PNS and rat CGRP were significantly greater in the arteries of OVX rats than in those of the sham-operated rats. OS-0689 inhibited the increase in vasorelaxation induced by both stimuli, whereas E2 had no effects. CONCLUSIONS: Ovariectomy not only decreases adrenergic function but also enhances CGRPergic function in rats' tail arteries. OS-0689 improves both impairments and thereby improves on rat hot flush. PMID- 16039419 TI - The role of cocaine in heroin-related deaths. Hypothesis on the interaction between heroin and cocaine. AB - In recent years, there has been an increase in cocaine-related deaths at the Department of Legal Medicine and Public Health of Pavia, probably reflecting the rising trend in cocaine use in Western Europe. Deaths from cocaine alone have increased from 6 cases in 1979-1991 (1.5% of drug-of-abuse deaths) to 13 in 1992 2002 (3.2%) and comparing the same periods, heroin-related deaths (HRDs) involving cocaine more than doubled from 8 (1.9%) to 22 (5.4%). In an attempt to investigate the role of cocaine in HRDs, acute narcotic death cases testing positive for cocaine use (blood cocaine or metabolite concentration >0.01 mg/l, COC+) were examined. Only cases from 1997 to 2001 were considered as in this period all data were obtained using the same analytical procedures (free morphine and total morphine by DPC Coat-A-Count radioimmunoassay before and after enzymatic hydrolysis, cocaine and metabolites in blood by SPE, TMS derivatization and gas chromatography-mass spectrometry (GC-MS)). The median, minimum and maximum concentrations of free morphine in blood (FM) and total morphine in blood (TM), urine (UM) and bile (BM) in the COC+ group (n = 9) were compared with those calculated in the group of "pure" HRDs (no other drugs detected in blood, COC-, n = 30). Differences among the medians in the two groups were statistically evaluated using the two-tailed Mann-Whitney U-Test. Statistical analysis was also carried out including in both groups cases with a blood alcohol concentration (BAC) > 20 mg/L (COC+, n = 19; COC-, n = 76). For the COC+ group, median TM was lower (0.32 mg/l versus 0.90 mg/l, P = 0.0214), median FM was lower, but not statistically significant (0.08 mg/l versus 0.28 mg/l, P = 0.1064), FM/TM ratio was similar (0.33 and 0.35), UM was also similar (21.0 mg/l and 18.0 mg/l), and BM was higher (90.0 mg/l versus 49.0 mg/l, P = 0.0268). Similar comparison results were obtained by repeating statistical analyses after including in the two groups cases with positive BAC. The picture observed for HRD cases involving cocaine is very different from what was previously observed for HRD cases involving ethanol [A. Polettini, A. Groppi, M. Montagna, The role of alcohol abuse in the etiology of heroin related deaths: evidence for pharmacokinetic interactions between heroin and alcohol, J. Anal. Toxicol. 23 (1999) 570-576], and updated with more recent data; in the high-ethanol (HE, BAC > 1000 mg/l) group, TM was lower than in the low-ethanol (LE, BAC 50% of the metaphyseal diameter) require bridging fixation besides percutaneous pin fixation. The use of this technique achieves the goal of surgical treatment of distal radius fractures: restoration of hand and wrist function through the restoration of alignment and articular surface congruity. PMID- 16039444 TI - Biomechanics and biology of plate fixation of distal radius fractures. AB - The fracture management principles of anatomic or near anatomic reduction, fracture stabilization, minimal operative trauma, and early joint motion are paramount in man-aging unstable distal radial fractures. The operative approach and plate selection should correlate with the fracture configuration. Plates have the advantages of providing secure fixation throughout the entire healing process without protruding wires or pins and allowing early and intensive forearm, wrist, and digital exercises. Disadvantages include additional operative trauma, including fragment devascularization; some additional risk of wrist stiffness; occasional tendon rupture; and at times, the need for plate removal. New developments in plate and screw design and operative strategies, fragment specific fixation, and plate strength have improved results with plate fixation. Fixed angle blades and locking screws and pegs enhance overall plate stability, support the articular surface of the distal radius, and are effective in fractures occurring in osteopenic bone. PMID- 16039445 TI - Dorsal plating for distal radius fractures. AB - A dorsal approach to the distal radius for comminuted intra-articular fractures affords excellent exposure of the joint surface. Dorsal plating can reliably buttress the joint, leading to low rates of arthrosis, but at the expense of wrist stiffness and high rates of extensor tendon problems. New, smaller implants may prove capable of supporting the joint with lower rates of extensor irritation. PMID- 16039446 TI - Volar plate fixation of distal radius fractures. AB - Volar fixed angle fixation may be considered as the beginning of a new era in restoring wrist function to patients with dorsally displaced distal radius fractures even in the face of comminuted or osteopenic bone. A thorough understanding of the anatomy of the wrist is a prerequisite when volarly approaching dorsally displaced distal radius fractures. The demonstration of the device theoretical and practical advantages requires an appreciation of the basics of working length, principles of plate stability, and the effect of cantilever bending. Volar fixed angle fixation successfully improves wrist function and significantly prevents the complications of the dorsal approach previously intractable to treatment. The current advantages, indications, clinical results, and complications of this new technology are being reviewed. PMID- 16039447 TI - Fragment-specific internal fixation of distal radius fractures. AB - The treatment of complex, intra-articular distal radius fractures can be challenging. Goals of treatment include the restoration of articular congruity and maintenance of a stable reduction to allow for early range-of-motion, prevention of arthrosis, and ultimately return of upper extremity function. Proper understanding of individual fracture patterns is paramount in achieving these goals and avoiding complications. Through the use of limited surgical incisions and low-profile,anatomically contoured implants, fragment-specific internal fixation provides a rational approach to the treatment of these complex injuries. PMID- 16039448 TI - Biomechanics and biology of external fixation of distal radius fractures. AB - External fixation is a versatile and useful tool for management of complex fractures. There is little to choose between the various types of commercially available fixators, and it is important to use one that allows the surgeon adequate versatility and follows sound biomechanical principles. Ligamentotaxis can be used effectively to reduce the most difficult fractures; however, over distraction and prolonged traction are harmful and should be avoided. Certain types of fractures do not respond to treatment with ligamentotaxis alone and require adjunctive treatment, such as limited internal fixation. A single K-wire significantly adds to the stability of fixation and should be considered in all cases. Understanding the basic mechanical principles and respect for pin-bone biology allow for successful use of external fixation with minimal complications. PMID- 16039450 TI - Nonbridging external fixation of intra-articular distal radius fractures. AB - New solutions to difficult problems are always welcome, but nonbridging external fixation of intra-articular fractures is still in its infancy. Multicenter clinical trials are necessary to deter-mine whether the superior results obtained with nonbridging fixation of extra-articular fractures can be duplicated. With further study and new fixator designs, it is anticipated that nonbridging external fixation of intra-articular distal radius fractures will become another viable treatment option. PMID- 16039449 TI - Non-spanning external fixation of the distal radius. AB - Non-spanning external fixation of the distal radius is a simple, reliable, and noninvasive technique to restore the anatomy in unstable extra-articular or minimal articular fractures of the distal radius. Rehabilitation is faster and more complete than with other comparable techniques. It is also a successful technique for stabilization of corrective osteotomy of the distal radius formal union. PMID- 16039451 TI - Combined internal and external fixation of distal radius fractures. AB - Combined internal and external fixation of distal radius fractures is used most commonly to treat injuries with joint surface or metaphyseal comminution. External fixation aids reduction intraoperatively and facilitates arthroscopic, per-cutaneous, or open manipulation of the fracture. Internal fixation maintains precise reduction of critical anatomy, principally the contour and orientation of the articular surface. Postoperatively the fixator functions as a neutralization device, preventing fracture collapse and decreasing the biomechanical demands on the internal fixation hardware. The combined technique exploits the benefits of both forms of fixation, allowing each to be used to full advantage in the treatment of complex distal radius fractures. PMID- 16039452 TI - Intra-articular distal radius fractures: the role of arthroscopy? AB - Arthroscopic assisted fixation of distal radius fractures offers several advantages. It allows for the evaluation of the articular reduction under a bright light and magnification. Particularly, rotation of the fracture fragments, which is difficult to judge under fluoroscopy, may be detected arthroscopically and corrected. Washing out fracture hematoma and debris potentially allows for improved range of motion, as shown by the studies of Doi and Stewart. Wrist arthroscopy also allows for detection and management of associated soft tissue injuries, which has been shown to occur with distal radius fractures. Some of the more severe interosseous ligament injuries can be diagnosed on plain or traction radiographs. Arthroscopy performed at the same time as fracture reduction, however, substantially increases the recognition of these injuries. In addition, it is wellknown that the management of acute interosseous ligament tears has a better prognosis when compared with chronic lesions. PMID- 16039453 TI - Treatment of injuries to the ulnar side of the wrist occurring with distal radial fractures. AB - We still do not know how to best treat the DRUJ condition that was recognized 200 years ago by Abraham Colles and later addressed in Frykman's classic thesis. To improve the outcome, we must recognize the differences be-tween osteoporotic and other fractures and understand the importance of ligament injuries,especially in patients under the osteoporotic age. However, our current problem is that neither the initial ligament injury nor the posttraumatic laxity is detectable with radiographic methods,which creates future challenges regarding diagnosis and treatment. We therefore have to critically analyze each fracture in each patient and be aware of the complexity of the entire injury to the wrist. PMID- 16039454 TI - Combined fractures of the scaphoid and distal radius: a revised treatment rationale using percutaneous and arthroscopic techniques. AB - Revision of the treatment rationale for combined fractures of the scaphoid and distal radius is based on evolution of treatment goals. The trend toward early recovery of hand function requires rigid fixation of both fractures before the start ofa hand therapy program. It is clear that prolonged immobilization of the scaphoid fracture jeopardizes early motion protocols for the distal radius. The fixation of unstable distal radius fractures with volar locking plates appears to offer the most stable construct to permit early motion. Evaluation, reduction, and fixation should be accomplished without disruption of the uninjured ligaments required for stable motion or the soft tissue envelope required for healing. Minimally invasive or percutaneous techniques are the meth-ods required. The tools needed are a clear understanding of anatomy, minifluoroscopic imaging units, and small-joint arthroscopy instruments. Many investigators advocate these techniques for scaphoid and distal radius fractures. It is only natural that these techniques should be used for these combined injuries. The key to success is a three-step process: (1)percutaneous reduction of the scaphoid fracture and provisional stabilization with a guide wire placed along its central axis, (2) percutaneous/arthroscopic reduction and rigid fixation of the distal radius fracture to permit early motion, and(3) fixation of the scaphoid fracture. This final step is accomplished by dorsal percutaneous implantation of a cannulated headless compression screw along the central scaphoid axis. Dorsal percutaneous fixation of scaphoid fractures with headless compression screws and rigid fixation of unstable distal radius fractures with a volar lock-ing plate system offer the most secure fixation. This small series suggests that the goals of early recovery of hand function can be accomplished using percutaneous/miniopen techniques for fracture reduction with rigid fixation and minimal risks. PMID- 16039455 TI - Nonunion of the distal radius. AB - Distal radius nonunion is either more common or more commonly recognized. The success of operative treatment to gain union seems to have improved along with improvements in operative fixation of fractures of the distal radius. Operative treatment to gain union or arthrodese the wrist can improve function and comfort with relatively few complications. PMID- 16039456 TI - Use of bone graft substitutes and bioactive materials in treatment of distal radius fractures. AB - Although autogenous bone graft has been shown to be useful in the treatment of distal radius fractures, the role of bone graft substitutes and the optimal replacement material remains unclear. Several products are commercially available, each with differing osteoconductive, osteoinductive, and structural properties. Indications and choice of graft substitute should be based on the needs of the individual case with regard to need for structural support, gap filling, or bone healing stimulation. Further comparative research will help clarify the indications and most appropriate material for a given fracture and clinical situation. PMID- 16039457 TI - Rehabilitation of distal radius fractures: a biomechanical guide. AB - Fracture healing and surgical decision making are not always predictable. The suggested protocols are intended to be flexible rather than rigid to be responsive to patient progress and the fracture site stability. A methodologic approach to the rehabilitation following a distal radius fracture, based on a knowledge of the biology of fracture healing and biomechanics of fixation, may preempt some of the pitfalls associated with distal radius fracture healing. PMID- 16039458 TI - Extra-articular distal radial fracture malunion. AB - With the increase in surgical options for the treatment of distal radius fractures, the authors anticipate that distal radial fracture malunions will be a less frequently seen problem. Nevertheless, they will still occur. Although patient selection has been weighted toward the younger patient, we believe that surgery should be based on patient activity level, functional needs, and disability related to the malunion. With advances in biotechnology and improved anesthetics, surgical intervention even in the older and osteopenic population is now more promising. Surgical intervention still requires appropriate patient selection, careful preoperative planning, and meticulous surgical technique. The appropriate surgical procedure should be tailored to the patient's symptoms, age, needs, and radiographic findings. PMID- 16039459 TI - Salvage of post-traumatic arthritis following distal radius fracture. AB - There are practical recommendations that can be drawn from the aforementioned results. Due to the minimal morbidity of the wrist denervation, patients with good but painful wrist motion following fracture of the distal radius should first be evaluated for wrist denervation unless formal resection of the dorsal interosseous nerve has clearly been included in the previous treatment. The evaluation is performed in a standardized manner before and after test infiltration of both interosseous nerves. This evaluation includes assessment of pain, strength, and working capacity. Whereas the grip strength often does not (cannot)increase more than 10% to 20%, the subjective pain relief can be remarkable, leading to higher repetition counts and increased dexterity. Inpatients with insufficient response to the anesthetic nerve blocks, other pain sources must be sought, especially on the ulnar side of the wrist. Patients with less than functional range are candidates for complete arthrodesis. A way for further evaluation with regard to the potential of partial and complete wrist arthrodesis is trial immobilization of the wrist in a light cast ora firm reinforced brace. Trial immobilization also allows anticipating the functional deficit from loss of range of wrist motion. Due to the still-unrestricted pronation and supination, ulnar-sid-ed wrist pain may persist and will need adequate follow-up adjunct treatment. Patients who have good pain relief but are not willing to completely lose their wrist motion should be evaluated fluoroscopically or receive lateral radiographs in full flexion and extension to measure their mid-carpal joint mobility and anticipate the potential residual motion after radiocarpal fusion. Patients without pain relief from test anesthesia, trial immobilization, and no apparent distal radioulnar joint pathology are poor candidates for further operative treatment.In evaluating different salvage procedures,among all diagnoses, painful arthritis following fracture of the distal radius is the most difficult to treat and yields the poorest results. Emphasis must therefore be on better initial fracture treatment and earlier secondary reconstructive interventions. The current salvage procedures must allow further improvement or alternatives must be developed. Prosthetic replacement merits serious consideration, especially when it can be adapted to the specific post-traumatic setting. This situation is not worse than rheumatoid arthritis because the clinician is dealing with healthy and strong intact bone stock, tendons, and ligaments,and most important, complete absence of a progressive disease. PMID- 16039460 TI - Long-term effect of corneal refractive excimer laser surgery. PMID- 16039461 TI - Intraocular pressure rise after phacoemulsification surgery in glaucoma patients. PMID- 16039464 TI - Preoperative assessment of cataract surgery patients. PMID- 16039465 TI - Preoperative LASIK screening: an evolving standard of care. PMID- 16039466 TI - Functional outcomes of acrylic IOLs in pediatric cataract surgery. PMID- 16039467 TI - Cortical removal simplified by J-cannula irrigation. PMID- 16039468 TI - Cataract ossea. PMID- 16039469 TI - Aggravation of proliferative diabetic retinopathy after LASIK. PMID- 16039470 TI - LASIK and the contrast sensitivity function. PMID- 16039471 TI - Cataract surgical problem. Cataract surgical problem. PMID- 16039481 TI - Modification of the Siepser slip-knot technique. AB - A new modification of the Siepser sliding knot technique allows the surgeon to create a locking knot when performing iris reconstruction or intraocular lens fixation to the iris. PMID- 16039482 TI - Secondary anterior capsulotomy in pseudophakic eyes. AB - This report describes a 1-handed technique and a 2-handed technique anterior capsulotomy in pseudophakic eyes. Each technique was used in a case of proliferative vitreoretinopathy accompanying anterior capsule contraction that occurred following cataract and intraocular lens (IOL) implantation. A capsulorhexis forceps and a stab knife were inserted into the anterior chamber, and the capsule contraction alleviated without extracting the IOL in both cases. Secondary anterior capsulotomy contributed to both the surgeon's visibility during intraocular surgery, including the subsequent vitreous surgery, and the patient's postoperative visual acuity. The transparency of the pupil region was restored. While the 1-handed technique involves the risk of adversely affecting Zinn's zonule, the 2-handed technique does not place a burden on Zinn's zonule and is thought to considerably improve the surgeon's visibility in subsequent vitreoretinal surgery. PMID- 16039483 TI - Excimer laser photorefractive keratectomy for hyperopia: 7.5-year follow-up. AB - PURPOSE: To report the results of a long-term prospective study to evaluate refractive stability and safety of hyperopic photorefractive keratectomy (H-PRK). SETTING: Department of Ophthalmology, St. Thomas' Hospital, London, United Kingdom. METHODS: Twenty-one patients (49%) (40 eyes) of cohort of 43 patients who participated in 1 of the first clinical trials of H-PRK were assessed at a mean follow-up of 90.7 months (range 62 to 106 months). The H-PRK was performed using a Summit Technology Apex Plus Excimer laser (Summit Technology, Inc.). The mean preoperative spherical equivalent refraction (SEQ) was +4.70 diopters (D) (range +2.00 to +7.50 D). Patients were allocated to 1 of 4 treatment groups based on their preoperative refraction and received 1 of the following spherical corrections: +1.50 D, +3.00 D, +4.50 D, or +6.00 D. RESULTS: At 7.5 years, the refractive correction remained stable with a mean difference in SEQ between 1 year and 7.5 years of +0.28 D. The mean manifest SEQ was +0.83 D (range +5.00 to 3.00 D). Sixty-seven percent of eyes having corrections of +1.50 D and +3.00 D were within +/-1.00 D of the predicted correction. Predictability was poorer with +4.50 D and +6.00 D corrections, with 40% of eyes within +/-1.00 D of that expected. An improvement in uncorrected near acuity was achieved in 35 eyes (87.5%), and 35 eyes (87.5%) showed an improvement in uncorrected distance acuity from preoperative levels. Best spectacle-corrected visual acuity (BSCVA) was unchanged or improved from preoperative values in 25 eyes (62.5%). Three eyes (8%) lost 2 lines of Snellen BSCVA, which in 2 cases was attributable to cataract formation. A peripheral ring of haze, 6.5 mm in diameter, appeared in most eyes. Its intensity was greatest at 6 months and then diminished with time. In 10 eyes (25%), remnants of the haze ring were evident at 7.5 years and subepithelial iron rings, 6.5 mm in diameter were evident in 26 eyes (70%). No patient complained of night-vision problems and no eye developed ectasia. CONCLUSIONS: In H-PRK, refractive stability achieved at 1 year was maintained up to 7.5 years with no evidence of hyperopic shift, diurnal fluctuation, or late regression. Peripheral corneal haze decreased with time but was still evident in a number of eyes at the last follow-up visit. PMID- 16039484 TI - Clinical comparison of 6 aberrometers. Part 1: Technical specifications. AB - PURPOSE: To provide a detailed assessment of the techniques, technical features, and practical use of 6 aberrometers made available to our institution from September 2002 to January 2004. SETTING: Department of Ophthalmology, University Hospital Antwerp, Antwerp, Belgium. METHODS: A number of technical and practical parameters are listed for the Visual Function Analyzer (Tracey), the OPD-scan (ARK 10000; Nidek), the Zywave (Bausch & Lomb), the WASCA (Carl Zeiss Meditec), the MultiSpot Hartmann-Shack device, and the Allegretto Wave Analyzer including working principles, data acquisition, aberrometer alignment, wavefront calculation, and data analysis. Operator and patient comfort as well as practical advantages and disadvantages are discussed. CONCLUSION: All devices met at least half the following parameters: alignment, correction for source wavelength, data averaging, measurement quality check, and inhibition of accommodation. PMID- 16039485 TI - Ocular higher-order aberrations in myopia and skiascopic wavefront repeatability. AB - PURPOSE: To study the distribution of ocular higher-order aberrations (HOAs) in a myopic population and to assess the repeatability of HOA measurements determined by a commercially available skiascopic wavefront sensor. SETTING: Department of Ophthalmology, Assaf Harofeh Medical Center, Zerifin, Israel. METHODS: Ocular HOAs were examined 3 times across a 6.0 mm naturally dilated pupil in 61 eyes using the Optical Path Difference (OPD)-scan wavefront aberrometer. Root-mean square (RMS) values of HOAs, total spherical aberration (TSA), total coma (TC), and total trefoil (TT) were analyzed. Correlation analysis was performed to assess the aberration symmetry between right and left eyes. The repeatability of the OPD-scan measurements was assessed by calculating Pearson r correlation coefficients between each pair of measurements and the interclass correlation coefficients between the 3 measurements of each score. RESULTS: The mean RMS values of HOAs, TSA, TC, and TT were 0.347 microm +/- 0.252 (SD), 0.120 +/- 0.174 microm, 0.165 +/- 0.168 microm, and 0.252 +/- 0.157 microm, respectively. The HOAs, TSA, TC, and TT changed slightly and not significantly with increasing refractive error (all P>.05). The RMS level of HOAs and TTR of the 3rd measurement was significantly different from the 1st and 2nd measurements (P<.05), with overall low correlation between the 3 measurements for the HOAs, TSA, TC, and TT. CONCLUSIONS: The ocular wavefront aberrations varied greatly from subject to subject. Ocular HOAs were not correlated with refractive error. The repeatability of HOAs measurements with the OPD-aberrometry was low. PMID- 16039486 TI - Clinical application of 11-deoxycortisol in visualizing prolapsed vitreous body after posterior capsule rupture in cataract surgery. AB - PURPOSE: To evaluate the clinical usefulness of 11-deoxycortisol, a precursor of cortisol in steroid metabolism, in visualizing the vitreous body in the anterior chamber after posterior capsule rupture during cataract surgery. SETTING: Department of Ophthalmology, Tsukuba University Hospital, Ibaraki, Japan. METHODS: Eight eyes had anterior vitrectomy during cataract surgery using this new technique. A suspension of 11-deoxycortisol was prepared by dilution in balanced salt solution without preservatives or emulsifying agents. After a posterior capsule rupture, the suspension was injected into the anterior chamber. The visual and anatomical outcomes of surgery were evaluated during a follow-up of 7 to 9 months. RESULTS: After the 11-deoxycortisol injection, the vitreous body that had prolapsed through the torn posterior capsule, with white particles entrapped on its surface, became clearly visible. Complete removal of the prolapsed vitreous body was achieved easily and safely using an anterior vitrectomy system. No postoperative complications related to the use of 11 deoxycortisol were observed. CONCLUSION: Injection of 11-deoxycortisol helped visualize the vitreous body in the anterior chamber after posterior capsule rupture. The technique minimized intraoperative and postoperative complications related to posterior capsule rupture and vitreous loss during cataract surgery [corrected] PMID- 16039487 TI - Temporal hinge laser in situ keratomileusis: maximizing treatable stromal bed area. AB - PURPOSE: To compare the treatable stromal bed area created by a temporal versus superior hinge laser in situ keratomileusis (LASIK) flap. SETTING: Assil Sinskey Eye Institute, Santa Monica, California, USA. METHODS: A randomized retrospective case review was performed of 2 subgroups (each with 12 eyes) of a cohort having LASIK for the correction of hyperopic astigmatism between August 2001 and March 2002. In 1 group, superior hinge LASIK was performed using the standard surgical technique for the 9.5 mm Hansatome suction ring. In the other group, the keratome was rotated 90 degrees such that a temporal hinge LASIK was performed with the 9.5 mm Hansatome suction ring. Corneal curvature (central mean keratometric value), flap diameter (arc length), and treatable stromal bed area were measured in each eye, and mean values were compared between the 2 groups. A treatable stromal bed area was defined as the circle centered over the entrance pupil, tangent to the hinge (devoid of hinge interference). RESULTS: The mean keratometry and mean flap diameter in the superior and temporal hinge groups measured 42.3 diopters (D) and 9.68 mm and 42.2 D and 9.69 mm, respectively. These differences were not significant (P=.882 for average keratometry and P=.943 for flap diameter). The mean treatable stromal bed area in the superior versus temporal hinge group measured 44.14 mm(2) and 62.25 mm(2), respectively. The difference between the 2 groups was highly significant (P<.0001). CONCLUSIONS: A large diameter, temporal hinge LASIK flap yielded a significantly greater area of treatable corneal stromal bed than a superior hinge flap. There are several clinical and theoretical advantages of optimizing stromal bed exposure for any given flap diameter. PMID- 16039488 TI - Clinical outcomes of phakic refractive lens in myopic and hyperopic eyes: 1-year results. AB - PURPOSE: To confirm the safety, efficacy, and predictability of the surgical correction of myopia and hyperopia with the phakic refractive lens (PRL) (Medennium Inc.). SETTING: St. Eriks Eye Hospital, Stockholm, Sweden. METHODS: This was a prospective clinical study of 20 eyes, 14 myopic and 6 hyperopic, that had PRL implantation at St. Eriks Eye Hospital from April to November 2002. Examinations were performed preoperatively and 1 day, 1 week, 3 months, and 1 year postoperatively. Follow-up included evaluation of the PRL rotation with retroillumination photography, evaluation of the distance between the PRL and the crystalline lens with Scheimpflug image, laser flare, endothelial cell count, uncorrected (UCVA) and best corrected (BCVA) visual acuity, residual refractive error, refractive stability, intraocular pressure, and induced cataract. RESULTS: Postoperatively, 11 eyes (55%) gained 1 or more lines, 5 eyes (25%) had no change, and 4 eyes (20%) lost 1 line of BCVA. No eye lost 2 or more lines. Mean UCVA was 0.87+/- 0.29 postoperatively. Laser flare values were highest 1 day after operation with normalization at 3 months and without changes at 1 year (P<.05). A rotation of 10 degrees or more was found in 15 eyes (75%) during the first year. The distance between the PRL and crystalline lens was considerably less at 1 year than at baseline (P<.05). There was no statistically significant endothelial cell loss induced by the PRL (P<.05). No induced cataract, glaucoma, or inflammation was observed. In 1 hyperopic eye, horizontal iris transillumination defects were noticed at 1 year. CONCLUSION: Safety and efficacy indexes were high at 1-year follow-up. The PRL rotated slightly in the posterior chamber. The distance between the PRL and the crystalline lens was considerably less at 1 year than at baseline. PMID- 16039489 TI - Conservation of corneal tissue with wavefront-guided laser in situ keratomileusis. AB - PURPOSE: To determine whether corneal tissue can be conserved with wavefront guided ablation compared to conventional surgery with a larger ablation zone for attempted prevention of glare and halo. SETTING: Department of Ophthalmology, Ilsan Paik Hospital, Inje University, Korea. METHODS: This prospective study was composed of 2 parts: First, 40 eyes of 20 patients were studied to determine whether a larger optical ablation could be beneficial in prevention of glare after conventional laser in situ keratomileusis (LASIK) surgery. One eye in each patient was treated with a 6.00 mm optical zone, and the other with a 6.25 mm optical zone. Second, 20 eyes of 10 patients with a higher-order root mean square (RMS) value of 0.3 or greater were evaluated to learn whether a wavefront-guided ablation could be as effective as a larger conventional optical ablation. One eye in each patient was treated by conventional LASIK surgery with a 6.25 mm optical zone, and the other eye was treated by wavefront-guided LASIK surgery with a 6.00 mm optical zone. All patients were analyzed with a WASCA analyzer (Carl Zeiss Meditec) preoperatively, and 1 month and 3 months after surgery. RESULTS: Higher order aberrations including coma, spherical aberration, and higher-order RMS were statistically significantly increased after conventional surgery with both a 6.00 mm and 6.25 mm optical zone. High-order aberrations including comaer- and spherical aberration after conventional surgery with 6.25 mm zone were statistically significantly increased. However, coma and higher-order RMS did not show a statistically significant increase between pre-LASIK and post-LASIK in wavefront-ablated eyes with a 6.00 mm optical zone. CONCLUSION: Wavefront ablation showed less increase of coma and higher-order RMS regardless of a smaller optical zone. This finding might provide a clinical clue for an advantage of wavefront-guided ablation from the standpoint of corneal tissue conservation. PMID- 16039490 TI - Thin flap laser in situ keratomileusis: flap dimensions with the Moria LSK-One manual microkeratome using the 100-microm head. AB - PURPOSE: To determine the predictability and consistency of corneal flap thickness, flap diameter, and hinge length with the modern 100 microm head of the Moria LSK-One manual microkeratome. SETTING: Private clinic, office-based practice. METHODS: Forty-two consecutive eyes with no previous surgery having thin flap laser in situ keratomileusis with the Moria LSK-One manual microkeratome had a new 100 microm (predicted flap thickness) head used for flap creation. Flap thickness was measured intraoperatively by subtraction ultrasound pachymetry (difference between central corneal thickness before flap cutting and residual stromal bed thickness after flap lifting). Vertical flap diameter and nasal hinge length were measured with calipers. RESULTS: Mean flap thickness was 107 microm +/- 14 (SD) (range 82 to 137 microm). Standard deviation for mean vertical flap diameter was +/-0.24 mm. The cord length of the nasal hinge was variable with a mean of 4.26 +/- 0.63 mm (range 3.12 to 5.75 mm) in length. Postoperatively, there were no slipped flaps, flap striae, diffuse lamellar keratitis, or epithelial defects; there was 1 epithelial slide. At 1 day, the visual acuity was 20/20 or better in 76% of eyes. CONCLUSIONS: The 100 microm head of the Moria LSK-One manual microkeratome cut a very predictable flap thickness and diameter but with variable length hinges. This flap thickness predictability was superior to that in other series with thicker intended flaps cut with mechanical microkeratomes and is comparable to that reported with the IntraLase FS femtosecond laser. Visual recovery was rapid, epithelial risks minimal, efficiency superior, and cost nominal relative to femtosecond laser technology. PMID- 16039491 TI - Macular thickness after cataract surgery with intracameral cefuroxime. AB - PURPOSE: To determine whether the use of prophylactic intracameral cefuroxime during phacoemulsification cataract surgery is associated with increased macular thickening in the postoperative period. SETTING: Hull and East Yorkshire Eye Hospital, Hull, United Kingdom. METHODS: In this prospective randomized double masked clinical study, the study group received 1 mg of intracameral cefuroxime and the control group received intracameral balanced salt solution only. Ocular coherence tomography was performed 5 (+/-1) weeks after surgery. RESULTS: Intracameral cefuroxime did not have a statistically significant effect on postoperative macular thickness compared with nonadministration of intracameral antibacterials. All patients in both groups achieved a postoperative best corrected visual acuity of 6/9 or better. CONCLUSION: Use of 1 mg of intracameral cefuroxime was not associated with increased macular thickness 4 weeks to 6 weeks postoperatively. PMID- 16039492 TI - Paired opposite clear corneal incisions to correct preexisting astigmatism in cataract patients. AB - PURPOSE: To evaluate the astigmatic correcting effect of paired opposite clear corneal incisions on steep axis in cataract patients. SETTING: Sligo General Hospital, Sligo, Ireland. METHODS: Fifteen eyes of 14 cataract patients with a mean age of 78.4 years +/- 6.38 (SD) (range 69 to 90 years) were recruited for the study. Inclusion criterion was topographic astigmatism of more than 2 diopters (D) in the cataractous eye. Preoperative refraction, autokeratometry, and topography were performed. The steep axis was marked before sub-Tenon's anesthesia was given. Paired 3-step self-sealing opposite clear corneal incisions were made 1 mm anterior to limbus on the steep axis with a 3.2 mm keratome. One incision was used for standard phacoemulsification, and the other was left unused for astigmatic correction. All the patients had day-case surgery. The first follow-up was at 1 month. Postoperative topography, keratometry, and refraction were performed on all patients. RESULTS: Mean preoperative and postoperative topographic corneal astigmatism were 3.26 +/- 1.03 D (range 2.30 to 5.80 D) and 2.02 +/- 1.04 D (range 0.20 to 4.00 D), respectively. Mean astigmatic correction was 1.23 +/- 0.49 D (range 0.30 to 2.20 D). Mean surgically induced astigmatism by vector analysis was 2.10 +/- 0.79 D (range 0.80 to 3.36 D). There were no incision-related complications. CONCLUSION: Paired opposite clear corneal incisions on the steep axis is a useful way to correct astigmatism in cataract patients, requiring no extra skill or instrumentation. PMID- 16039493 TI - Comparison of dynamic digital pupillometry and static measurements of pupil size in determining scotopic pupil size before refractive surgery. AB - PURPOSE: To compare 2 digital infrared pupillometers. SETTING: Anterior Segment Service, Moorfields Eye Hospital, London, England. METHODS: Eighty-eight eyes of 44 healthy subjects were recruited into the study. The scotopic pupil size was measured with 2 instruments, a digital pupillometer (Procyon) and a Hartman-Shack wavefront aberrometer (Visx WaveScan). Agreement between the instruments was assessed. RESULTS: There were 21 men and 23 women with a mean age 38.1 years +/- 9.2 (SD) (range 23 to 62 years). The mean scotopic pupil diameter was significantly larger with the WaveScan (6.61 +/- 0.92 mm) than with the Procyon pupillometer (6.40 +/- 0.90 mm), P<.001. CONCLUSIONS: There was good agreement in measurement of scotopic pupil size between the two instruments. The larger pupil diameter found with the Visx WaveScan than with the Procyon digital pupillometer probably represents the different illumination level used by each instrument, the effects of accommodation, and target fogging. PMID- 16039494 TI - Steepening of corneal curvature with contraction of the ciliary muscle. AB - PURPOSE: To measure the changes of corneal curvature during contraction of the ciliary muscle. SETTING: Department of Ophthalmology, St. Luke's International Hospital, Tokyo, Japan. METHODS: Twenty-eight eyes of 14 healthy volunteers under 40 years old were enrolled in this prospective randomized controlled study and divided into pilocarpine and control groups. Intraocular pressure (IOP), pupil diameter, and corneal topography were measured before and 40 minutes after instillation of topical pilocarpine 4% or balanced salt solution. Corneal topography was analyzed for the mean ring-power of Placido rings 1 through 25, average corneal power (ACP), and for spherical equivalent, regular astigmatism, asymmetry, and high-order irregularity by Fourier analysis. RESULTS: Pilocarpine had no effect on IOP, but it did cause a significant decrease in mean pupil diameter. Simultaneously, pilocarpine increased the mean ring powers for Placido rings 1 through 4 and the ACP (+0.13 diopters (D) +/- 0.17 [SD]; P=.017). By Fourier analysis, the mean spherical component for the central 3.0 mm of the cornea increased in the pilocarpine group (+0.08 +/- 0.15 D; P=.020). There were no changes in components of regular astigmatism, asymmetry, and high-order irregularity. CONCLUSIONS: The central cornea steepened in curvature and increased in power owing to contraction of the ciliary muscle. The results suggest that changes in corneal curvature increase refractive power during accommodation. PMID- 16039495 TI - Effect of tilt of 2 acrylic intraocular lenses on high-order aberrations. AB - PURPOSE: To investigate the high-order aberrations (HoAs) in pseudophakia between 2 foldable acrylic intraocular lenses (IOL) with differences in the tilt of IOL. SETTING: Department of Ophthalmology, Nara Medical University, Nara, Japan. METHOD: Sixty-four eyes that had uncomplicated phacoemulsification and IOL implantation were assigned to 1 of 2 groups: Acryfold 60BB (HOYA [n=30 eyes]) and AcrySof MA60AC (Alcon [n=34 eyes]). All patients had best corrected visual acuity better than 20/25. High-order aberrations were measured using the Hartmann-Shack aberrometer at 4.0 mm and 6.0 mm wavefront aperture diameters, and the IOL tilt and decentration were measured using Scheimpflug videophotography. RESULTS: There was a significant difference in the IOL tilt (60BB=2.22 degrees +/- 1.44 (SD), 60AC=3.18 +/- 1.84 degrees; P=.041, Mann-Whitney U test), but no difference in IOL decentration (P>.05). In the cornea, there were no differences between the 2 groups in HoAs at 4.0 mm and 6.0 mm aperture diameters (P>.05, Mann-Whitney U test). In the whole eye, there was no difference between the 2 groups in spherical-like aberrations, coma-like aberrations, and total aberrations at the 4.0 mm and 6.0 mm aperture diameters (P>.05). The compensation of the internal optics with the 60BB group was better than with the 60AC group for coma-like (P=.037) and total aberrations (P=.010) at 6.0 mm aperture diameter. CONCLUSION: The smaller tilt of the IOL induced more compensation for the coma-like and total aberrations at the 6.0 mm aperture diameter. The IOL tilt should be small, especially in a large pupil, with regard to HoAs. PMID- 16039496 TI - Randomized comparative clinical study of cryoanalgesia versus topical anesthesia in clear corneal phacoemulsification. AB - PURPOSE: To compare intraoperative pain scores and objective stress signs during clear corneal phacoemulsification under cryoanalgesia and topical anesthesia. SETTING: Hospital Ramon y Cajal, Madrid, and Hospital Universitario Nuestra Sra. de la Candelaria, Tenerife, Canary Islands, Spain. METHODS: Eighty-two patients were randomized to have phacoemulsification under cryoanalgesia or topical anesthesia. Uncooperative patients and those with shallow anterior chamber and small pupils were excluded. In case of breakthrough pain during the surgery, a supplemental anesthesia protocol was established. Each patient was asked to grade the severity of pain on a 4-point scale (verbal description score; 0=none, 1=little, 2=some, or 3=much). Immediately after surgery, the general discomfort and pain were evaluated. Surgeon stress was evaluated during surgery. A comparison of the 2 groups was performed using a statistical analysis of variance. RESULTS: Supplemental anesthesia was required in 1 patient in each group. A total of 95.23% of patients would repeat the same technique under cryoanalgesia versus 97.5% under topical anesthesia. Similar pain levels and surgical stress scores were noted in both groups. CONCLUSIONS: Cryoanalgesia clear corneal phacoemulsification was safe with an acceptable level of pain. It induced a physiological stress response to that of topical anesthesia (blood pressure and heart rate). Cryoanalgesia was preferred over topical anesthesia by some patients. It is a suitable technique for anesthetic allergy cases. PMID- 16039497 TI - Decrease in complications during cataract surgery with the use of a silicone tipped irrigation/aspiration instrument. AB - PURPOSE: To compare rates of vitreous loss during cataract surgery when a silicone-tipped irrigation/aspiration (I/A) instrument is used for cortex removal as opposed to a metal tip. SETTING: Parkland Memorial Hospital, Dallas, Texas, USA. METHODS: A retrospective chart review of all patients who had cataract extraction by phacoemulsification by third-year ophthalmology residents between September 2000 and February 2004 was conducted. A silicone-tipped I/A instrument was used to remove cortex for all surgeries beginning in September 2002, whereas a metal I/A tip was used previously. RESULTS: Of the 1072 cases performed with a metal I/A tip, there were 13 cases of vitreous loss during cortex removal (rate 1.2%) and 26% of all vitreous loss during that time occurred during cortex removal. Of the 805 cases performed with a silicone I/A tip, there was a single case (0.1%) of vitreous loss during cortex removal (P=.004); only 4% of all vitreous loss during that time occurred during cortex removal (P=.011). CONCLUSION: The overall incidence of vitreous loss during cortex removal and the ratio of vitreous loss during cortex removal to all cases of vitreous loss were significantly decreased using the silicone-tipped I/A instrument. PMID- 16039498 TI - Comparison of clear corneal phacoemulsification combined with 25-gauge transconjunctival sutureless vitrectomy and standard 20-gauge vitrectomy for patients with cataract and vitreoretinal diseases. AB - PURPOSE: To compare effectiveness and outcomes of clear corneal phacoemulsification combined with 25-gauge transconjunctival sutureless vitrectomy and standard 20-gauge vitrectomy system for patients with clinically significant cataract and vitreoretinal diseases. SETTING: Department of Ophthalmology, Tri-Service General Hospital, Taipei, Taiwan, Republic of China. METHODS: Clear corneal phacoemulsification combined with 25-gauge transconjunctival sutureless vitrectomy was performed in the study group (15 eyes). Patients in the control group (15 eyes) had combined surgery with clear corneal phacoemulsification and the existing 20-gauge vitrectomy system, providing a comparison between 2 groups with regard to preoperative and postoperative best corrected visual acuity (BCVA), intraocular pressure, and intraoperative and postoperative complications. The additional outcome measurements in both groups were duration of the surgical procedures, time to return to preoperative corneal clarity, time to achieve stable vision, and subjective ocular irritation postoperatively. RESULTS: Postoperatively, BCVA improved in 12 eyes (80%) in the study group and in 11 eyes (73.3%) in the control group. Mean operative time for opening and closing the sclerotomy was significantly greater in the control group than in the study group. Time to return to preoperative corneal clarity and time to achieve stable vision showed no significant difference between the groups. Compared with the control group, postoperative ocular irritation in patients in the study group was considerably reduced. CONCLUSIONS: Combined sutureless surgery needed shorter setup time for sclerotomy and caused less postoperative ocular irritation than combined surgery with 20-gauge vitrectomy. Therefore, this type of procedure would be a good option for selected cases with cataract and vitreoretinal diseases. PMID- 16039499 TI - Novel tissue adhesives to secure laser in situ keratomileusis flaps. AB - PURPOSE: To evaluate 2 novel biodendrimer tissue adhesives in sealing and securing laser in situ keratomileusis (LASIK) flaps. SETTING: Duke University Eye Center, Durham, North Carolina, USA. METHODS: Laser in situ keratomileusis flaps were created in 10 human eye-bank eyes using the Hansatome microkeratome system (Bausch & Lomb). These eyes were divided into 2 groups. Flaps in the first group (n=4) were secured with a laser-activated biodendrimer adhesive along the flap edge. In the second group (n=6), the flaps were secured with a self-gelling dendritic adhesive. Dry Merocel sponges (Medtronic Solan) were used to test the strength of flap adherence in both groups. Further testing was performed in the second group. The hinges of these flaps were cut with a scalpel blade and fluorescein dye was injected under the flap to observe potential dye leakage along the flap edge. RESULTS: Laser in situ keratomileusis flaps sealed with both adhesives were secure with no flap dislocation. There was no leakage of fluorescein dye observed in the second group. Both adhesives were easy to apply, clear when dry, and had a soft rubbery consistency. CONCLUSIONS: Two novel biodendrimer adhesives successfully sealed and secured LASIK flaps. These adhesives may prove to be an effective alternative for treating LASIK flap complications such as epithelial ingrowth or flap dislocation. PMID- 16039500 TI - Hyaluronic acid combined with mannitol to improve protection against free-radical endothelial damage: experimental model. AB - PURPOSE: To evaluate the protective properties of combined sodium hyaluronate 2% and mannitol 0.5% (Visiol) on the corneal endothelium in the presence of oxidative stress induced by hydrogen peroxide (H(2)O(2)). SETTING: Instituto Oftalmologico de Alicante, Universidad Miguel Hernandez, Alicante, Spain. METHODS: This was an exploratory randomized controlled parallel-group, masked assessor study of 3 sodium hyaluronate-based ophthalmic viscosurgical devices (OVDs): Visiol, Healon (sodium hyaluronate 1%), and Viscoat (sodium hyaluronate 3%-chondroitin sodium 4%). The OVDs were tested for protective effects on the endothelium following oxidative stress induced by H(2)O(2) at increased concentrations: control (lactated Ringer's solution), 1 mM, 10 mM, and 100 mM. Groups without OVD were used as controls at the same concentrations of peroxide. Each animal received the same treatment in both eyes (10 eyes per group). Endothelial cell lesion was assessed using the Janus green photometry absorbance technique. RESULTS: At 10 mM peroxide concentration, the value of endothelial cell lesion was significantly lower in the Visiol (16.8%, P=.0056), Healon (22.2%, P=.0302), and Viscoat (21.6%, P=.0336) groups than in the control group (29.4%, no OVD). There was a trend in favor of Visiol to more efficiently reduce cell lesions of the endothelium, than Healon (P=.055) and Viscoat (P=.1013). Values of endothelial cell lesion at peroxide concentrations of 1 mM and 100 mM showed the same trends than those observed at 10 mM. CONCLUSIONS: All of the OVDs tested efficiently reduced endothelial lesions against free radicals compared with the control group in which no OVD was used. The following sequence for the efficacy of endothelial cell protection was established: Visiol>Viscoat>Healon>no OVD. PMID- 16039501 TI - Growth factor deposition in anterior subcapsular cataract. AB - PURPOSE: To characterize immunohistochemically the distribution of growth factors and extracellular matrix (ECM) components in an anterior subcapsular cataract (ASC) and to determine the role of growth factors in the development of ASC. SETTING: Department of Ophthalmology, Wakayama Medical University, Wakayama, Japan. METHODS: During cataract surgery in 22 patients, anterior capsules with an ASC were obtained. Sections of each specimen were immunostained with a panel of antibodies against ECM components, growth factors, cytoskeletal components, and signal transduction-related molecules. RESULTS: Collagen types I, V, and VI; fibronectin; fibrillin-1; and latent transforming growth factor beta binding protein-1 (LTBP-1) were localized to the ECM in ASC tissues. Collagen IV was localized to the ECM and the capsule. Lens epithelial cells (LECs) were positive for alpha-smooth muscle actin (alphaSMA). Lens epithelial cells and ECM stained for transforming growth factor beta2 (TGFbeta2) and TGFbeta3 in all samples, but TGFbeta1 latency-associated peptide (TGFbeta1-LAP) were detected in some samples. Fibroblast growth factor-2 (FGF-2) and hepatocyte growth factor-alpha (HGF-alpha) were localized to the ECM. Lens epithelial cells with nuclear staining for Erk-1, the mitogen-activated protein kinase (MAP kinase) cascade-related molecule, and Smad3, 1 of the Smad family members involving TGFbeta signaling, were detected. CONCLUSIONS: Matrix components (ie, collagen types, fibronectin, fibrillin-1), as well as growth factors such as TGFbeta1-LAP, TGFbeta2, TGFbeta3, FGF-2, and HGF alpha, were detected in ASC. Fibrillin-1 might serve as a repository for TGFbetas. These growth factors may modulate the phenotypic alteration and behavior of LECs. The MAP kinase cascade and TGFbeta signaling are both activated in LECs in ASC. PMID- 16039502 TI - Transcriptional activation in lens epithelial cells following an ocular blunt trauma. AB - PURPOSE: To determine whether an ocular blunt trauma activates anterior ocular segment (cornea and lens) by examining the expression patterns of c-fos and c-jun mRNAs in these tissues of an eye of adult rat following a blunt trauma. SETTING: Department of Ophthalmology, Wakayama Medical University School of Medicine, Kimiidera, Wakayama, Japan. METHODS: Adult Wistar rats (n=36) were generally anesthetized by ether inhalation. One eye was hit with an iron sphere (30 gram) that fell to the eye from 1 m. After the procedure, the animals were killed and the affected eye was enucleated at 15, 30, 60, 120, and 180 minutes. In situ hybridization using radiolabeled oligoprobes was used to detect mRNAs of c-fos and c-jun in tissue. RESULTS: The c-fos and c-jun mRNAs were not detected in the epithelium of uninjured cornea and lens by in situ hybridization. The mRNAs for c fos and c-jun were then detected in corneal epithelium from 15 to 60 minutes posttreatment, and were no longer observed thereafter. In lens epithelium, mRNA for c-fos or c-jun were transiently detected from 15 to 60 minutes or 30 minutes posttreatment, respectively. CONCLUSION: The c-fos and c-jun mRNAs were transiently expressed in corneal and lens epithelial cells after blunt trauma. Ocular blunt trauma activates corneal and lens epithelial cells without apparent corneal ablation or direct injury in the lens epithelium. Such activation in lens epithelium might be involved in cataractogenesis. PMID- 16039503 TI - Experimental evaluation of the Corneal Concept 360 intraocular lens with the Miyake-Apple view. AB - PURPOSE: To evaluate a new single-piece, hydrophilic acrylic intraocular lens (IOL) designed to keep the anterior capsule away from the IOL optic, create the effect of a capsular tension ring, and prevent posterior capsule opacification (PCO). SETTING: John A. Moran Eye Center, University of Utah, Salt Lake City, Utah, USA. MATERIALS AND METHODS: Four Concept 360 IOLs (Corneal Laboratoire) were implanted in 4 human cadaver eyes of various sizes (Miyake-Apple technique). Lens centration, ovalization of the capsular bag and capsulorhexis opening, distance between the anterior optic surface and the anterior capsule and between the posterior optic surface and the posterior capsule, as well as presence of capsular striae were evaluated. The capsular bag was stained with trypan blue, and the ophthalmic viscosurgical device (OVD) was stained with fluorescein to enhance visualization. RESULTS: In each eye, the lens conformed to the capsular bag with precise centration. No ovalization or distortion of the capsular bag or the capsulorhexis and no posterior capsule striae were observed. Measurements of the capsulorhexis diameter before and after IOL implantation were the same. The anterior capsule remained at distance from the anterior optic surface for 360 degrees, and tight contact between the posterior optic edge and the posterior capsule was observed. CONCLUSIONS: Use of the Miyake-Apple technique is useful in the evaluation of new IOL designs in experimental studies. Design features of the Concept 360 IOL are likely to decrease the outcome of postoperative capsular bag opacification. Long-term clinical studies are necessary to confirm these findings. PMID- 16039504 TI - Unexpected outcomes associated with laser in situ keratomileusis: ptosis, anisocoria, and "curing" of exotropia. AB - Laser in situ keratomileusis (LASIK) is a common and popular procedure that is generally associated with excellent results and few complications. We present 3 cases of unusual LASIK-related complications. In the first case, the patient developed anisocoria after otherwise unremarkable surgery. In the second case, the patient developed mild eyelid ptosis. In the third case, the patient's exotropia resolved after mild refractive overcorrection. In each case, the visual outcome was excellent and no further intervention was necessary. PMID- 16039505 TI - Suprachoroidal hemorrhage during cataract surgery in a vitrectomized eye. AB - We report a case of surprachoroidal hemorrhage (SCH) that occurred during cataract surgery in a previously vitrectomized eye. The only sign of SCH was a progressive shadow obscuring the red reflex. There was no increase in intraocular pressure (IOP), shallowing of the anterior chamber, or iris prolapse. Postoperatively, the SCH gradually resolved without complications, leaving a series of subretinal pigmentary lines. Surgeons should be aware that the signs normally expected in SCH may not develop in vitrectomized eyes. Obscuration of the red reflex may be the only sign of the hemorrhage, and when this happens, it may be prudent to keep the IOP elevated and suture the incision at the end of the procedure. PMID- 16039506 TI - Clinical manifestation and pathologic finding of unilateral acute hydrops after bilateral laser in situ keratomileusis. AB - We present a case of unilateral acute hydrops that developed in 25-year-old man 6 years after bilateral laser in situ keratomileusis (LASIK) for the correction of 2.75 diopters (D) of myopia. Preoperative corneal thickness using ultrasound pachymetry were 556 microm in the right eye and 554 microm in the left eye. Preoperative corneal topography of both eyes showed an asymmetric bow-tie pattern with inferior steepening. Forty-seven months after LASIK, the left eye developed astigmatism and enhancement excimer ablation was performed. Sixteen months after enhancement, a definite sign of keratectasia was detected in the left eye. Twenty six months after enhancement, acute hydrops occurred in the left eye. The interface of the LASIK wound was separated and filled with aqueous humor. Penetrating keratoplasty was performed to avoid perforation. The keratocytes at the edge between the peripheral flap and remaining stroma showed mixoid degeneration and edematous change suggesting dying cells. PMID- 16039507 TI - Lamellar interface fluid accumulation following traumatic corneal perforation and laser in situ keratomileusis. AB - A 41-year-old man with myopic astigmatism had laser in situ keratomileusis (LASIK) in each eye in April 2002. Ten months later, he sustained a central perforating corneal injury to the right eye. One day following repair of the corneal wound, he presented with diffuse corneal epithelial microcystic edema, lamellar interface fluid accumulation, and 20/400 visual acuity. Additional sutures were placed to close a presumed posterior wound gape with complete resolution of the corneal edema and lamellar interface fluid collection. One year later, his best corrected visual acuity measured 20/20+ in the right eye. This case is the first to document lamellar interface fluid accumulation following LASIK owing to traumatic disruption of the corneal endothelium. PMID- 16039508 TI - Postcataract endophthalmitis caused by Mycobacterium goodii. AB - This is the first reported case of delayed-onset postcataract endophthalmitis caused by Mycobacterium goodii, confirmed by multiple cultures. The patient had uneventful cataract removal by phacoemulsification with implantation of a posterior chamber intraocular lens (IOL). One month later, he developed redness, pain, a hypopyon, and a decrease in visual acuity to finger counting in the affected eye. A vitreous biopsy was performed for suspected endophthalmitis; culture results showed rapidly growing bacteria identified by DNA sequencing as Mycobacterium goodii. The eye ultimately required IOL explantation but had a good final outcome with 20/40 uncorrected vision. Mycobacterium goodii is a rapidly growing Mycobacterium with isolates more commonly occurring in cellulitis, osteomyelitis, and respiratory disease. Single-drug antibiotic therapy for nonocular disease is often sufficient if the correct antimicrobial agent is used. The newer fluoroquinolones are promising against rapidly growing Mycobacterium. PMID- 16039509 TI - Late subluxation of an ICL. PMID- 16039510 TI - Sumatriptan: a potent drug for management of corneal lesion pain. PMID- 16039511 TI - Dynamics of indocyanine green dye in the anterior chamber. PMID- 16039512 TI - Intraoperative retinoscopy: a way to achieve emmetropia in cataract surgery. PMID- 16039513 TI - Intraoperative fracture of AMO Clariflex silicone posterior chamber IOL. PMID- 16039514 TI - Bilateral endophthalmitis after simultaneous bilateral cataract surgery. PMID- 16039515 TI - Intracorneal cotton fiber in radial keratotomy incision for 20 years. PMID- 16039517 TI - Broad institute: bringing genomics to real-world medicine. PMID- 16039518 TI - Designing effective hybrid toxins. AB - A study of a well-designed androgen-mustard conjugate provides evidence supporting a novel mechanism for its selective toxicity in androgen-receptor positive cancer cells. This represents a solid step forward on the path toward effective hybrid toxins for targeted cancer therapy. PMID- 16039519 TI - Bacterial crowd control with iron. AB - Bacterial biofilms account for more than 80% of human infections. Hergenrother and coworkers report in this issue that high concentrations of iron salts can block the formation of these bacterial communities; this represents an attractive new method for biofilm control. PMID- 16039520 TI - On the road to bioremediation of "dioxin". AB - Mohammadi and Sylvestre report engineering of a dioxygenase to create an enzyme that attacks dibenzofuran in the lateral position. Subsequent oxidation and a second dioxygenation produced ring-open products. All metabolites were unambiguously identified by 1H-NMR. This new pathway targets degradation of chlorinated dibenzofurans. PMID- 16039521 TI - Iteratively acting glycosyltransferases involved in the hexasaccharide biosynthesis of landomycin A. AB - Detailed studies on the biosynthesis of the hexasaccharide side chain of landomycin A, produced by S. cyanogenus S136, revealed the function of each glycosyltransferase gene of the biosynthetic gene cluster. Analyses of generated mutants as well as feeding experiments allowed us to determine that LanGT2 and LanGT3 catalyze the attachment of one sugar, whereas LanGT1 and LanGT4 attach two sugars during landomycin A biosynthesis. The generation of a lanZ2 deletion mutant provided evidence that LanZ2 is controlling the elongation of the saccharide side chain. PMID- 16039522 TI - Chemical approaches to define the structure-activity relationship of heparin-like glycosaminoglycans. AB - Heparin, the drug of choice for the prevention and treatment of thromboembolic disorders, has been shown to interact with many proteins. Despite its widespread medical use, little is known about the precise sequences that interact with specific proteins. The minimum heparin binding sequence for FGF1 and FGF2 necessary to promote signaling was investigated. A characteristic pentasaccharide sequence, DEFGH, is required to accelerate the inhibition of thrombin and factor Xa in the blood-coagulation cascade. The first synthetic heparin pentasaccharide drug has been approved in Europe and the US and is sold under the trade name Arixtra. Other oligosaccharides with different composition are under clinical investigation. The enormous interest in the assembly of heparin oligosaccharides will stimulate the development of new synthetic approaches. Heparin oligosaccharide-synthesis automation similar to that of DNA or peptide synthesis will play an important role. PMID- 16039523 TI - Functional dissection of sRNA translational regulators by nonhomologous random recombination and in vivo selection. AB - Small nontranslated RNAs (sRNAs) regulate a variety of biological processes. DsrA and OxyS are two E. coli sRNAs that regulate the translation of rpoS, which encodes a protein sigma factor. Due to their structural complexity, the functional dissection of sRNAs solely by designing and assaying mutants can be challenging. Here, we present a complementary approach to the study of functional RNAs, in which highly diversified RNA libraries are generated by nonhomologous random recombination (NRR) and processed efficiently by in vivo selections that link RNA activities to cell survival. When applied to DsrA and OxyS, this approach rapidly identified essential and nonessential regions of both sRNAs. Resulting hypotheses about DsrA and OxyS structure-function relationships were tested and further refined experimentally. Our findings demonstrate an efficient, unbiased approach to the functional dissection of nucleic acids. PMID- 16039524 TI - Phosphoproteomic analysis with a solid-phase capture-release-tag approach. AB - A comprehensive study of global phosphorylation events in biological systems is critical. We report a chemistry-based capture-release-tag method for isolation of complex phospho-Ser/Thr-containing peptides by liquid beta-elimination combined with solid-phase Michael addition. The free thiol groups of 6-(mercapto acetylamino)-hexanoic acid functionalized resin are used as immobilized Michael donors to capture dehydro-serine/threonine peptides. After an acid-mediated release step, phospho-peptides are labeled with a 6-(2-mercapto-acetylamine) hexanoic amide tag at phosphorylated sites. We applied this method to analyze the phosphorylation status of microtubule-associated proteins. We find that a CDK5 substrate microtubule-associated protein 2 (MAP2) is phosphorylated on residues that are within a homologous region of Tau. The chemical method corroborates previous results and suggests that Tau and MAP2 may contain a CDK5 phosphorylation motif. PMID- 16039525 TI - Disruption of gene expression and induction of apoptosis in prostate cancer cells by a DNA-damaging agent tethered to an androgen receptor ligand. AB - The goal of our work was the design of DNA-damaging agents that disrupt both DNA repair and signaling pathways in prostate tumor cells. A DNA alkylator (N,N-bis-2 chloroethyl aniline) was linked to a steroid ligand (17beta-hyroxy-estra Delta(4(5),9(10))-3-one) to produce a complex molecule (11beta-dichloro) that forms DNA adducts that bind the androgen receptor (AR). We speculated that DNA adducts in an AR-DNA adduct complex would be camouflaged from DNA repair proteins that would otherwise remove the adducts in prostate cancer cells. Furthermore, transcription dependent on the AR would be antagonized by AR redistribution to sites distant from AR-driven promoters. The anticancer potential of 11beta dichloro was demonstrated against prostate cancer cells in vitro and in vivo. 11beta-dichloro induces a unique pattern of gene disruption, induces apoptosis in apoptosis-resistant cells, and shows promising anticancer activity in animals. PMID- 16039526 TI - Iron salts perturb biofilm formation and disrupt existing biofilms of Pseudomonas aeruginosa. AB - Bacterial biofilms are thought to aid in the survivability of a variety of intractable infections in humans. Specifically, biofilm production in Pseudomonas aeruginosa has been shown to play a significant role in chronic infection of cystic fibrosis (CF) patients. Unfortunately, no clinically effective inhibitors of biofilm formation are available. A rapid screen of 4509 compounds for nonantibiotic biofilm inhibitors in Pseudomonas aeruginosa PA14 was executed in 384-well plates. Among those compounds, ferric ammonium citrate inhibited biofilm formation in a dose-dependent manner; other iron salts functioned similarly. In addition to biofilm inhibition in static culture, pregrown biofilms could be disrupted and cleared by switching to iron-rich media in flow-chamber experiments. Furthermore, P. aeruginosa strains taken from the sputum of 20 CF patients showed a similar response to elevated iron levels. Previous expression profiling analyses demonstrated that high levels of iron repress the expression of genes whose products are essential for scavenging iron and that expression of these genes is critical for virulence. Our results, combined with existing transcriptional-profiling data, now indicate that elevated iron concentrations repress the expression of certain genes essential for biofilm production in P. aeruginosa. PMID- 16039527 TI - Inhibition of hIAPP amyloid-fibril formation and apoptotic cell death by a designed hIAPP amyloid- core-containing hexapeptide. AB - The pathogenesis of type II diabetes is associated with the aggregation of the 37 residue human islet amyloid polypeptide (hIAPP) into cytotoxic beta sheet aggregates and fibrils. We have recently shown that introduction of two N-methyl rests in the beta sheet- and amyloid-core-containing sequence hIAPP(22-27), or NFGAIL converted this amyloidogenic and cytotoxic sequence into nonamyloidogenic and noncytotoxic NF(N-Me)GA(N-Me)IL. Here, we show that NF(N-Me)GA(N-Me)IL is able to bind with high-affinity full-length hIAPP and to inhibit its fibrillogenesis. NF(N-Me)GA(N-Me)IL also inhibits hIAPP-mediated apoptotic beta cell death. By contrast, unmodified NFGAIL does not inhibit hIAPP amyloidogenesis and cytotoxicity, suggesting that N-methylation conferred on NFGAIL the properties of NF(N-Me)GA(N-Me)IL. These results support the concept that rational N-methylation of hIAPP amyloid-core sequences may be a valuable strategy to design pancreatic-amyloid diagnostics and therapeutics for type II diabetes. PMID- 16039528 TI - Defining Cdk5 ligand chemical space with small molecule inhibitors of tau phosphorylation. AB - Cyclin-dependent kinase 5 (Cdk5) is widely viewed as a possible target for a wide variety of neurological disorders. One pathological role attributed to Cdk5 is the abnormal phosphorylation of tau that may lead to the neuronal inclusions known as neurofibrillary tangles. A high through-put screen for inhibitors of Cdk5-mediated phosphorylation of tau resulted in three compounds with distinct mechanisms of action. One compound is competitive with ATP and has a high affinity for the Cdk5 ATP binding pocket. The second compound also competes with ATP, is noncompetitive with tau, and (uniquely among this class of inhibitors) displaces adjacent amino acid residues to make room for the nitrophenyl group. A third compound did not compete with ATP, but did compete with tau at low concentrations of tau. The SAR and charge optimization derived from cocrystals of the two ATP competitors along with cocrystals of three other ATP competitors map out the importance of filling and properly charging different regions of the ATP binding pocket. Taken together, this analysis shows how the structure of Cdk5 constrains the space of potential inhibitors and reveals a pocket unfilled in all of the structures. These leads could be a starting point for structure-based drug design of more potent and selective inhibitors. PMID- 16039529 TI - Shift to Pseudomonic acid B production in P. fluorescens NCIMB10586 by mutation of mupirocin tailoring genes mupO, mupU, mupV, and macpE. AB - Mupirocin, a polyketide-derived antibiotic from Pseudomonas fluorescens NCIMB10586, is a mixture of pseudomonic acids (PA) that target isoleucyl-tRNA synthase. The mup gene cluster encodes both type I polyketide synthases and monofunctional enzymes that should play a role during the conversion of the product of the polyketide synthase into the active antibiotic (tailoring). By in frame deletion analysis of selected tailoring open-reading frames we show that mupQ, mupS, mupT, and mupW are essential for mupirocin production, whereas mupO, mupU, mupV, and macpE are essential for production of PA-A but not PA-B. Therefore, PA-B is not simply produced by hydroxylation of PA-A but is either a precursor of PA-A or a shunt product. In the mupW mutant, a new metabolite lacking the tetrahydropyran ring is produced, implicating mupW in oxidation of the 16-methyl group. PMID- 16039530 TI - Resolving the profile of metabolites generated during oxidation of dibenzofuran and chlorodibenzofurans by the biphenyl catabolic pathway enzymes. AB - Although the metabolism of dibenzofuran by the biphenyl catabolic enzymes had been inferred in previous reports, the metabolic pattern has never been determined unambiguously. In this work, we describe the evolved biphenyl dioxygenase (BPDO) RR41 that exhibits a higher turnover rate of metabolism toward dibenzofuran and chlorodibenzofurans than the parental Burkholderia xenovorans LB400 BPDO. We used RR41 BPDO to identify unambiguously the metabolites produced from the oxygenation of dibenzofuran by LB400 BPDO, and we evaluated their further metabolism by the biphenyl catabolic pathway enzymes of strain LB400. RR41 BPDO was obtained by saturation mutagenesis of targeted amino acid residues. I335F336N338I341L409 of LB400 BphA were replaced by A335M336Q338V341F409 in RR41 BphA. Data confirm the critical role played by these amino acid residues for substrate specificity and regiospecificity. PMID- 16039531 TI - Ligand-regulated peptides: a general approach for modulating protein-peptide interactions with small molecules. AB - We engineered a novel ligand-regulated peptide (LiRP) system where the binding activity of intracellular peptides is controlled by a cell-permeable small molecule. In the absence of ligand, peptides expressed as fusions in an FKBP peptide-FRB-GST LiRP scaffold protein are free to interact with target proteins. In the presence of the ligand rapamycin, or the nonimmunosuppressive rapamycin derivative AP23102, the scaffold protein undergoes a conformational change that prevents the interaction of the peptide with the target protein. The modular design of the scaffold enables the creation of LiRPs through rational design or selection from combinatorial peptide libraries. Using these methods, we identified LiRPs that interact with three independent targets: retinoblastoma protein, c-Src, and the AMP-activated protein kinase. The LiRP system should provide a general method to temporally and spatially regulate protein function in cells and organisms. PMID- 16039532 TI - Molecular imaging. PMID- 16039533 TI - SPECT imaging of herpes simplex virus type1 thymidine kinase gene expression by [(123)I]FIAU(1). AB - RATIONALE AND OBJECTIVES: Introduction of suicide genes, such as herpes simplex virus type1 thymidine kinase (HSV1-tk), in tumor cells has provided a useful method for tumor gene therapy. Several L-nucleosides, such as Lamivudine (3TC) and Clevudine (L-FMAU), have been successfully tested as high-potency antiviral agents. To investigate the potential differences between D- and L-isomers of nucleosides, [(125/123)I]-2'-fluoro-2'-deoxy-1beta-D/L-arabino-furanosy-5-iodo uracil (D/L-FIAU) have been synthesized and evaluated as potential SPECT agents for imaging HSV1-tk gene expression. MATERIALS AND METHODS: [(125/123)I]D- and L FIAU were prepared by iododestannylation of the respective tin precursors with (125/123)I-sodium iodide. In vitro cell uptake studies were performed by incubation of [(125)I]D- and L-FIAU in RG2 cells expressing HSV1-tk (RG2TK+). In vivo studies including biodistribution and SPECT were performed in RG2TK+ and RG2TK- tumor-bearing nude mice using [(123)I]D- and L-FIAU. RESULTS: Cell uptake and biodistribution studies indicated that [(125/123)I]L-FIAU did not show any high accumulation (sensitivity) or uptake ratios (selectivity) in HSV1-TK positive (RG2TK+) tumors as compared to control tumors. In contrast, [(125/123)I]D-FIAU displayed both sensitivity and selectivity to RG2TK+ tumors. The selective in vivo accumulation of [(123)I]D-FIAU increased with time and the tumor uptake ratios (RG2TK+/RG2TK-) for 2, 4, and 24 hours averaged 6.2, 22.7, and 58.8, respectively. High-resolution SPECT of four nude tumor-bearing mice demonstrated a very high uptake of [(123)I]D-FIAU in the RG2TK+ tumor, while no significant tracer accumulation was observed in the RG2TK- tumor and other organs. CONCLUSION: The data suggest that only the D-isomer of [(123)I]FIAU is useful for imaging HSV1-tk gene expression in mice by high-resolution SPECT imaging. PMID- 16039534 TI - Computer-aided detection applied to breast MRI: assessment of CAD-generated enhancement and tumor sizes in breast cancers before and after neoadjuvant chemotherapy. AB - RATIONALE AND OBJECTIVES: MRI has shown promise in assessing breast cancer patients undergoing neoadjuvant chemotherapy. Computer-aided detection (CAD) for MRI can automatically display tumor enhancement parameters. This study was performed to determine the utility of CAD applied to breast MRI in this patient population. MATERIALS AND METHODS: Fifteen patients with 16 newly diagnosed locally advanced breast cancers were evaluated with MRI before and after neoadjuvant chemotherapy. CAD assessments, including presence or absence of significant enhancement, enhancement profiles, and maximum sizes, were recorded. Pre-chemotherapy and post-chemotherapy enhancement profiles were compared. Sizes were compared to those measured by the radiologist and at final pathology. RESULTS: Prior to chemotherapy, all tumors demonstrated CAD-assessed significant enhancement. Following chemotherapy, 7/16 tumors showed no residual significant enhancement, but all had residual disease at pathology. In those patients with residual enhancement, comparison of the post-chemotherapy to pre-chemotherapy CAD enhancement profiles showed a significant decrease in percentage of washout enhancement (P = 0.0147) in patients with less than 5 mm of residual microscopic disease. Radiologist-measured tumor sizes demonstrated better correlation with sizes at pathology (r = 0.60) than did CAD-generated tumor sizes (r = 0.32). CONCLUSION: CAD may be helpful in assessing changes in MRI enhancement profiles of tumors following chemotherapy. However, CAD-assessed significant enhancement following chemotherapy can be falsely negative for residual malignancy, and CAD tumor sizes are less accurate than those measured by the radiologist in predicting size of residual malignancy. CAD may complement but should not replace the radiologist's assessment of tumors in this patient population. PMID- 16039535 TI - Semiautomatic nonrigid registration for the prostate and pelvic MR volumes. AB - RATIONALE AND OBJECTIVES: Three-dimensional (3D) nonrigid image registration for potential applications in prostate cancer treatment and interventional magnetic resonance (iMRI) imaging-guided therapies were investigated. MATERIALS AND METHODS: An almost fully automated 3D nonrigid registration algorithm using mutual information and a thin plate spline (TPS) transformation for MR images of the prostate and pelvis were created and evaluated. In the first step, an automatic rigid body registration with special features was used to capture the global transformation. In the second step, local feature points (FPs) were registered using mutual information. An operator entered only five FPs located at the prostate center, left and right hip joints, and left and right distal femurs. The program automatically determined and optimized other FPs at the external pelvic skin surface and along the femurs. More than 600 control points were used to establish a TPS transformation for deformation of the pelvic region and prostate. Ten volume pairs were acquired from three volunteers in the diagnostic (supine) and treatment positions (supine with legs raised). RESULTS: Various visualization techniques showed that warping rectified the significant pelvic misalignment by the rigid-body method. Gray-value measures of registration quality, including mutual information, correlation coefficient, and intensity difference, all improved with warping. The distance between prostate 3D centroids was 0.7 +/- 0.2 mm after warping compared with 4.9 +/- 3.4 mm with rigid-body registration. CONCLUSION: Semiautomatic nonrigid registration works better than rigid-body registration when patient position is changed greatly between acquisitions. It could be a useful tool for many applications in the management of prostate. PMID- 16039536 TI - Research articles published in clinical radiology journals: trend of contribution from different countries. AB - RATIONALE AND OBJECTIVES: To determine different countries' trend of contribution to clinical radiology journals and its relationship with impact factor. MATERIALS AND METHODS: All the journals, which publish articles on clinical radiology, were selected from the category of Radiology and Nuclear Medicine group of journals, and articles published in these journals between 1991 and 2000 were searched for the authors' affiliation using the Medline database. Then, share of research output of the top-ranking 20 countries was determined along with the trend over time. Also, the relationship of different countries' contribution with the impact factor of journals was examined by cross-sectional time-series linear model. RESULTS: Of total articles (38,359), the United States' share for the selected journals in clinical radiology was 43.2% (16,582 articles) and ranked top in the world, followed by the United Kingdom (9.9%) and Japan (8.0%). The recent increase in the share was statistically significant for Japan, France, Germany, Italy, South Korea, Turkey, Spain, Switzerland, Austria, and China. On the other hand, the United States, United Kingdom, and Canada showed a significant negative trend. Among the top-ranking 10 countries, the US contribution was significantly higher to journals with high-impact factors, whereas the opposite was true for France. CONCLUSION: The United States, United Kingdom, and Canada showed a negative trend over the last decade in terms of proportion of contribution of articles to the clinical radiology journals. However, only the United States published more articles in high-impact factor journals. PMID- 16039538 TI - Region-of-interest-based analysis of clustered BOLD MRI data in experimental arthritis. AB - RATIONALE AND OBJECTIVES: BOLD MRI provides functional information based on minimal changes. Problems inherent in data processing of the very low signal-to noise-ratio of BOLD experiments have created obstacles for validation of certain techniques using standard strength-field MR scanners. Measures of diagnostic accuracy of clustered data are directly related to the reading parameters used to define regions-of-interest (ROIs). Our primary aim was to determine the combination of ROI-related reading parameters that provides highest accuracy for discrimination of presence or absence of arthritis in acute and subacute stages of the disease using paired comparisons of BOLD MRI data. MATERIALS AND METHODS: Six male New Zealand white rabbits were injected with albumin into one knee and saline into the contralateral knee, 3 animals had albumin injected into only one of the knees, 2 had saline injected into one of the knees, and 3 animals were not injected. The rabbits' knees underwent BOLD MRI on days 1 and 28 after induction of arthritis, except for the knees of 3 animals (albumin- vs saline-injected knees, n = 2 animals; saline- vs noninjected knees, n = 1 animal) that died before expected and had only the first MRI examination done. Percentage of activated voxels and differences in on-and-off signal intensities were the BOLD MRI methods applied. Data were analyzed using anatomic-driven small ROI, voxel chaser-driven small ROI and anatomic-driven large ROI techniques. RESULTS: Diagnostic areas-under-the curve (AUCs) were obtained only for acute arthritis and only when percentage of activated voxels was used. Low threshold, positive voxel activations and small ROIs generated the largest AUCs (AUC +/- SE, .911 +/- .092, P = .014) using either anatomic-driven or voxel-chaser-driven techniques. A sensitivity analysis confirmed the importance of threshold as a parameter for analysis. CONCLUSION: Low threshold, positive voxel activations and small ROIs constituted the set of reading parameters that provided the most accurate BOLD MRI results. PMID- 16039537 TI - Effects of lesion conspicuity on visual search in mammogram reading. AB - RATIONALE AND OBJECTIVES: The goal of mammography screening is to detect breast cancer at early stages, but because of the complexity of the breast parenchyma and the variability of signs of the disease, many cancers go unreported when initially visible on the mammogram. We compared the visual search strategy used by experienced mammographers in a case set where they examined both the mammogram in which a malignant mass was discovered at screening mammography and the most recent prior mammogram. MATERIALS AND METHODS: Four experienced mammographers participated in this experiment. They read a case set of 20 two-view mammograms, of which 15 contained a malignant mass and 5 were lesion-free, in two trials. For each of the cancer cases, two versions were shown to the observers: the one in which the cancer was reported in the clinical practice, called the "current" mammograms, and the most recent prior. Each trial had a balanced mix of current and prior mammograms. In addition, the same set of lesion-free cases was shown to the observers in both trials. The eye movements of the observers were tracked, and visual search parameters such as time to hit the location of the malignant mass, dwell, and mean pupil size in the location of the cancer were collected. Statistical analyses were used to determine whether there were differences between the current and prior mammograms. RESULTS: A total of 66% of the malignant masses in the current mammograms and 57% in the priors attracted some amount of visual attention. From these, 71% yielded a report on the current mammograms, but only 40% on the priors. In the cases where the observer saw the malignant mass, they did so within 2 seconds of image display, regardless of whether the mammogram was current or prior. CONCLUSION: Most unreported malignant masses attracted some amount of visual attention, but it was in the processing of the information extracted in the location of the lesion that most errors occurred. In our experiment, approximately 70% of the total time used by the observers for visual scan of the cases was spent gathering information to corroborate the hypothesis already formed by the radiologist. PMID- 16039539 TI - Evaluating hormone therapy-associated increases in breast density comparison between reported and simultaneous assignment of BI-RADS categories, visual assessment, and quantitative analysis. AB - RATIONALE AND OBJECTIVES: Changes in breast density, which are commonly associated with hormone replacement therapy (HRT) use, may imply changes in breast cancer risk. This study explores the ability of different methods to detect hormone replacement therapy (HRT)-associated increases in breast density. MATERIALS AND METHODS: Between 1997 and 2001, 51 postmenopausal women were reported to have HRT-associated increases in breast density at our institution. Twenty postmenopausal women not reported to have an increase in density during the same period were selected as controls. Mammograms from date of report and earlier comparison were used. Breast Imaging Reporting and Data System (BI-RADS) density categories from both dates were obtained from the mammography report. Mammograms were reviewed at separate time points and density changes evaluated by assigning BI-RADS density categories, visual assessment, and computer-assisted quantitative analysis. RESULTS: Mammogram reports were not available for two patients. The remaining 49 women with reported HRT increases in density were included. Reported BI-RADS categories resulted in detection of 57%, simultaneous BI-RADS assignment in 61%, visual assessment in 100%, and quantitative assessment in 94% of women with HRT-associated increases in density. Reported BI-RADS category change was the only method that resulted in false-positive increases in density for control patients. Minimal HRT associated increases in density were the most difficult to detect, with 90% of these 21 cases not detected by simultaneous BI-RADS category assignment and 3 cases not detected by quantitative methods when defined as an increase of at least 5%. CONCLUSION: Visual and quantitative assessment best identified women with HRT-associated increases in density, including those with minimal increases. Simultaneous assignment of BI RADS categories was considerably better than use of reported BI-RADS categories. This information may be helpful in guiding research design of studies evaluating changes in density from the HRT use. PMID- 16039540 TI - Computerized texture analysis of mammographic parenchymal patterns of digitized mammograms. AB - RATIONALE AND OBJECTIVES: Mammographic density and parenchymal patterns have been shown to be related to the risk of developing breast cancer. Thus, computerized texture analysis of breast parenchymal patterns on mammograms may be useful in assessing breast cancer risk. MATERIALS AND METHODS: A comparative evaluation was conducted of various computer-extracted texture features of mammographic parenchymal patterns of women with BRCA1/BRCA2 gene mutations and those of women at low risk of developing breast cancer. Mammograms from 172 subjects (30 women with either the BRCA1 or BRCA2 gene mutation and 142 low-risk women) were analyzed. Computerized texture features were extracted from regions-of-interest to assess the mammographic parenchymal patterns in the images. Receiver operating characteristic analysis was used to assess the performance of these features in the task of distinguishing between the two groups of women. RESULTS: Quantitative texture analysis on digitized mammograms demonstrated that gene-mutation carriers and low-risk women have different mammographic parenchymal patterns. Gene mutation carriers presented with parenchymal patterns that were denser, coarser, and lower in contrast than those of the low-risk group. For the gene-mutation carriers, their mammographic patterns appear to contain less high-frequency component as indicated by higher coarseness values, lower fractal dimensions, and smaller edge gradients, which yielded corresponding A(z) values of 0.79, 0.84, and 0.78, respectively, in the task of distinguishing between gene-mutation carriers and the low-risk group with the entire dataset. The contrast measure calculated from co-occurrence matrix method, which describes local image variation, yielded an A(z) value of 0.86 in distinguishing between the two groups of women. CONCLUSION: Computerized texture analysis of mammograms provides radiographic descriptors of mammographic parenchymal patterns. The computer extracted features may be useful for identifying women at high risk for breast cancer and for monitoring the treatment of breast cancer patients. PMID- 16039541 TI - Toward objective quantification of perfusion-weighted computed tomography in subarachnoid hemorrhage: quantification of symmetry and automated delineation of vascular territories. AB - RATIONALE AND OBJECTIVES: Perfusion-weighted computed tomography (CTP) is a relatively recent innovation that estimates a value for cerebral blood flow (CBF) using a series of axial head CT images tracking the time course of a signal from an intravenous contrast bolus. MATERIALS AND METHODS: CTP images were obtained using a standard imaging protocol and were analyzed using commercially available software. A novel computer-based method was used for objective quantification of side-to-side asymmetries of CBF values calculated from CTP images. RESULTS: Our method corrects for the inherent variability of the CTP methodology seen in the subarachnoid hemorrhage (SAH) patient population to potentially aid in the diagnosis of cerebral vasospasm (CVS). This method analyzes and quantifies side to-side asymmetry of CBF and presents relative differences in a construct termed a Relative Difference Map (RDM). To further automate this process, we have developed a unique methodology that enables a computer to delineate vascular territories within a brain image, regardless of the size and shape of the brain. CONCLUSIONS: While both the quantification of image symmetry using RDMs and the automated assignment of vascular territories were initially designed for the analysis of CTP images, it is likely that they will be useful in a variety of applications. PMID- 16039542 TI - Predicting radiology resident errors in diagnosis of cervical spine fractures. AB - RATIONALE AND OBJECTIVES: Our objective was to identify factors associated with resident errors of cervical spine fractures to enable targeted education. MATERIALS AND METHODS: We performed a retrospective cohort study of consecutive cases of after-hours resident interpreted cervical spine fractures over 27 months at a single level 1 academic trauma center. The outcome measure was appropriate identification of all fractures by the resident. Potential predictors of resident error or discrepancy were identified from chart review and included: age, gender; fracture location/pattern (upper/lower cervical spine, occipital condyle, C1 ring, dens, C2 pars, vertebral body, posterior column, lateral mass, transverse process); consecutive and nonconsecutive additional fractures; radiologist distracting factors (number of noncervical spine injuries); number of noncervical spine studies performed. Risk ratios with confidence intervals were calculated for categorical variables using epidemiological 2 x 2 tables, and for continuous variables using difference of means. RESULTS: There were 59 errors among 492 cervical spine fractures in a total of 327 patients. Fifty-seven of the errors were on computed tomography and 2 errors were on radiographs. Upper cervical fractures were significantly more likely to have been errors than lower cervical fractures: risk ratio (RR) of 2.2 (confidence intervals (CI) 1.3, 3.5; P = .001). Occipital condyle fractures were more likely to have been discrepant: RR = 2.2 (CI 1.3, 3.9; P = .006). Dens fractures were also significantly more likely to have been discrepant: RR = 2.0 (CI 1.0, 3.8; P = .05). Other potential predictors were not associated with significantly increased risk. CONCLUSION: Upper cervical spine fractures, in particular occipital condyle and dens fractures were significantly associated with an increased relative risk of resident missing or misinterpreting the fracture. These findings suggest that resident education should focus in particular on upper cervical spine injuries, occipital condyle, and dens fractures. The methods used in this study could also be applied to other imaging modalities and anatomic regions in the future to target resident education to more challenging areas. PMID- 16039543 TI - A remedy for resident evaluation and remediation. PMID- 16039544 TI - Receiver operating characteristic analysis: a primer. PMID- 16039545 TI - Teleradiology accreditation. PMID- 16039548 TI - Development of an enzyme-linked immunosorbent assay for podophyllotoxin. AB - A competitive indirect enzyme-linked immunosorbent assay (ciELISA) for podophyllotoxin was developed by using polyclonal antibody, and its suitability for the determination of this analyte in spiked water samples was studied. To avoid antibody production to the linker, the succinoyl-podophyllotoxin (hapten) mimicking the analyte was synthesized and conjugated with the carrier proteins bovine serum albumin (BSA) and ovalbumin (OVA) by mixed anhydride reaction (MAR) and active ester method (AEM). Polyclonal antibodies raised against Hapten-BSA(1) synthesized by MAR and Hapten-BSA(2) by AEM were screened and selected for the ciELISA. One set of antibodies from the rabbit M4440 immunized with Hapten- BSA(1) showed an I(50) value of 2.21 microg/mL with a detection limit of 0.12 microg/mL, and the other set from the rabbit M4469 immunized with Hapten-BSA(2) had an I(50) value of 0.7897 microg/mL with a detection limit of 0.0056 microg/mL. This assay showed the cross-reactivities with the structurally closely related compounds. Recoveries from the podophyllotoxin-fortified tap water in the assay were in the range of 72--115%. A good correlation between podophyllotoxin concentration measured by the ELISA and HPLC (R(2)=0.9924, Y=1.195X-0.257) was obtained from linear regression analysis. These results indicate that the ELISA could be a convenient and supplemental analytical tool for monitoring podophyllotoxin and its analogues in waters without previous extraction or cleanup. PMID- 16039549 TI - Assessment of TNFalpha contribution to the functional up-regulation of kinin B(1) receptors in the mouse paw after treatment with LPS. AB - It has been widely demonstrated that LPS is able to induce kinin B(1) receptor up regulation throughout several models of inflammation. Using an in-vivo system in which LPS was administered systemically, we assessed the participation of the pro inflammatory cytokine TNFalpha in the functional up-regulation of B(1) receptors in the mouse paw. Systemic treatment with LPS (10 microg/animal, i.v. 24 h before) resulted in a marked increase (about 5-fold) in the mouse paw edema induced by the selective B(1) receptor agonist des-Arg(9)-BK (50 nmol/paw) in both Swiss and C57/BL6 mice. The up-regulation of des-Arg(9)-BK-caused edema following LPS treatment was found to be greatly diminished in TNFalpha p55(-/-) receptor knockout mice. In addition, the paw edema evoked by des-Arg(9)-BK was significantly reduced when mice received the anti-TNFalpha antibody (100 [corrected] microg/kg, i.v.) 5 min before the LPS treatment. A similar inhibition of B(1) receptor-mediated paw edema was observed when mice were treated with thalidomide (30 mg/kg, s.c.) [corrected] a drug known for reducing TNFalpha synthesis, 5 min prior to LPS administration. ELISA experiment [corrected] revealed that TNFalpha serum levels were maximal at 1 h following LPS systemic treatment. Taken together, the present results suggest that the early production of the pro-inflammatory cytokine TNFalpha is probably responsible for driving the sequence of events involved in the functional up-regulation of B(1) receptors in the mouse paw. PMID- 16039550 TI - A soy diet accelerates renal damage in autoimmune MRL/Mp-lpr/lpr mice. AB - Isoflavones, which are phytoestrogens present in large quantities in soy and soy derived products, have estrogenic activity, inhibit protein tyrosine kinase, and exert other effects in the human body. Thus, the recent spread of soy consumption in Western populations emphasizes the need to more fully understand the potential effects in the body, especially in abnormal immune conditions. In the present study, the influence of a soy diet on lupus disease in MRL/Mp-lpr/lpr (MRL/lpr) mice was investigated. Weanling female MRL/lpr mice (4 weeks) were fed a soy diet (20% soybean protein and 5% soybean oil). The soy diet exacerbated renal damage; findings in this mouse strain included accelerated proteinuria, elevated serum creatinine concentrations, and reduced creatinine clearance. No effects were detected, however, in C3H/HeN mice, which have the same H-2(k) genetic background as MRL/lpr mice do. A tendency toward an increase in thymus weight and proliferation of T cells in spleen and B cells in lymph nodes were found at the age of 16 weeks. These findings indicate that a soy diet, in comparison with a casein diet, significantly exacerbates the clinical course of this autoimmune disease. Further research on the mechanism of this effect of soy-rich diets is needed, and isoflavone supplementation for systemic lupus erythematosus patients should be carefully reevaluated. PMID- 16039551 TI - Chemically modified tetracyclines induce apoptosis in cultured mast cells. AB - Chemically modified tetracyclines are a group of non-antimicrobial tetracycline derivatives, which possess antiinflammatory, anticollagenolytic and antiproliferative properties. Here we studied the effects of four different chemically modified tetracyclines (CMT-1, CMT-3, CMT-8 and CMT-308) on proliferation and viability of cultured mouse and human mast cells. All studied CMTs (25 microM) effectively inhibited the viability and proliferation of human mast cell line (HMC-1) cells and mouse bone marrow derived mast cells (mBMMCs), as judged by trypan blue exclusion and by incorporation of [(3)H]thymidine. The antiproliferative effect of CMTs was not dependent on the stimulating growth factor, i.e. CMTs inhibited both IL-3 and c-kit ligand-induced proliferation of mBMMCs. The reduced viability of mast cells was due to induction of apoptosis, as indicated by the increased amount of apoptotic nucleosomes and the appearance of TUNEL positive cells in the presence of CMTs. The induction of apoptosis was further confirmed by showing that CMT-3 induces activation of caspase-3 and caspase-9 in HMC-1 cells. Additionally, CMT-3 induced downregulation of the expression of antiapoptotic Bcl-2 protein in HMC-1 cells. Compared to doxycycline, the antiproliferative and proapoptotic effects of different CMTs were clearly more pronounced. Of the studied CMTs, CMT-3 and CMT-8 appeared to be the most potent inhibitors of mast cell proliferation and survival. The present results show that CMTs have an antiproliferative and proapoptotic effect on both malignant and non-malignant mast cells. In conclusion, CMTs could offer a novel means to treat disorders with inappropriate expansion of mast cells, such as rheumatoid arthritis and systemic mast cell diseases. PMID- 16039552 TI - Modulation of acute inflammation by targeting glycosaminoglycan-cytokine interactions. AB - Glycosaminoglycans (GAGs) located on cellular membranes and the extracellular matrix (ECM) are able to interact with chemokines and pro-inflammatory cytokines, leading to local cytokine/chemokine accumulation. The tissue-bound cytokines/chemokines function in promoting leukocyte migration and activation, contributing to local inflammation. Hence, targeting of GAG-cytokine interactions may provide an avenue for the attenuation of inflammatory responses. A cationic peptide (MC2) derived from the heparin-binding sequence of mouse IFN-gamma was previously shown by our laboratory to delay allograft rejection in an animal model. In order to further investigate potential anti-inflammatory properties of the MC2 peptide, we have studied its activity in an acute peritoneal inflammation model. Groups of C57Bl/6 mice were injected intraperitoneally with either ConA or thioglycollate and treated with saline (control), the MC2 peptide or two control cationic peptides, poly-l-lysine (PLL) and poly-l-arginine (PLA). Treatment with the MC2 peptide, but not PLA or PLL, resulted in statistically significant reductions in total cell numbers, concentration of total proteins and concentrations of pro-inflammatory cytokines (TNFalpha, IL-6 or IL-1 beta) in peritoneal lavage fluids, without alterations to the qualitative cellular composition of the exudate. These results suggest that targeting GAG-cytokine interaction is a viable approach to reduce inflammation. PMID- 16039553 TI - Prior exposure to the carrier regulates rat immune responses to a conjugate vaccine. AB - Small vaccine antigens including peptides generally need to be linked to larger molecules or carriers in order to induce high levels of immune responses. The potential of unwanted immune responses to the carriers represents a major drawback for the conjugated vaccines. The carriers could also regulate the immune responses to the haptens and these effects need to be prevented in order to achieve adequate responses to the vaccines. We examined means to reduce the unwanted reactions to the carrier. For this purpose, we investigated whether prior exposure of rats to a human IgG(1) (hIgG(1)) carrier would affect their subsequent responses to an OVA peptide (i.e., OVA(173-196)). Prior exposure to the hIgG(1) carrier did not affect the T cell responses to the peptide antigen. However, IgG(1) Ab responses to the peptide antigen were enhanced while IgE Abs were reduced. These results show that responses to the hapten are not systematically relevant to the carrier pre-immunization and that the conjugate could achieve desired responses by selective immune responses suppression. Such models of vaccines with enhanced anti-hapten responses and reduced levels of potentially harmful responses could be of great interest for the development of new immune therapies. PMID- 16039554 TI - Aqueous extract of Salvia miltiorrhiza attenuates increased endothelial permeability induced by tumor necrosis factor-alpha. AB - Salvia miltiorrhiza Bunge, a traditional Chinese herbal medicine, is often used for prevention and treatment of cardiovascular disorders such as atherosclerosis. To understand its mechanism of pharmacological action, its effects on endothelial monolayer permeability are studied. The present study demonstrated that extract of S. miltiorrhiza (ESM) and its major ingredients, Danshensu (DSS) and salvianolic acid B (Sal B), inhibited tumor necrosis factor (TNF-alpha) induced endothelial permeability, whereas the other major ingredient, protocatechualdehyde, was ineffective. ESM, DSS and Sal B also repressed expression of vascular endothelial growth factor (VEGF) and extracellular signal regulated kinase (ERK) activation in TNF-alpha induced HUVEC cells. Furthermore, it was found that ESM attenuated the disorganization of vascular endothelial (VE) cadherin induced by TNF-alpha. The effect of ESM on TNF-alpha induced endothelial permeability and redistribution of VE-cadherin is likely due to a reduction of VEGF protein expression as a result of modulation of the ERK signaling pathway. Endothelial cell hyperpermeability is implicated in inflammation and subsequent ischemic reperfusion injury and atherosclerosis. Data from this study suggest that one of the mechanisms S. miltiorrhiza exerts its pharmacological effect is through its modulation of endothelial cell permeability. PMID- 16039555 TI - Effects of Turkish pollen and propolis extracts on respiratory burst for K-562 cell lines. AB - Bee-collected pollen and propolis are apicultural products which are composed of nutritionally valuable substances and contain considerable amounts of polyphenol substances which may act as potent antioxidants. We wanted to show if respiratory burst within a cancer cell lines could be influenced when incubated with pollen and propolis extracts or not. Pollen and propolis extracts at concentrations of 50, 25, 12.5 and 0 mg/ml were prepared by dimethyl sulfoxide (DMSO). K-562 cell cultures and mononuclear cell (MNC) cultures prepared from a peripheral blood sample to serve as control cells were incubated with extracts for 24 h. Determination of respiratory burst was carried out by intracellular dichlorofluorescein (DCFH) test by using flow-cytometric fluorescence analysis. While about 90% and 66% fluorescence was detected at zero concentrations for both K-562 and MNC cultures, fluorescence positivity decreased (between 3.8% and 11.8%) as concentrations of both propolis and pollen extracts increased for K-562 cell culture, but unchanged (between 20% and 83%) for MNC culture. It was concluded that pollen and propolis extracts inhibit respiratory burst within cancer cell lines probably by their antioxidant potentials. PMID- 16039556 TI - The bad seed in Alzheimer's disease. AB - In this issue of Neuron, McGowan et al. report on a new mouse model of amyloid deposition as occurs in Alzheimer's disease. Unlike previous models in which overexpression of the amyloid precursor protein results in amyloid plaque formation, McGowan et al. have produced mice that overexpress only Abeta40 or Abeta42 and prove that Abeta42 is critical for the formation of amyloid deposits in vivo. PMID- 16039558 TI - Noncommutativity of eye rotations and the half-angle rule. AB - In order to produce kinematically efficient behavior when dealing with the noncommutativity of rotations, the oculomotor system has developed strategies such as the half-angle rule. In this issue of Neuron, Ghasia and Angelaki demonstrate that during smooth pursuit eye movements the half-angle rule is implemented by the mechanical properties of the eye plant. PMID- 16039557 TI - Cortical maps: where theory meets experiments. AB - Primary visual cortex (V1) has remarkably systematic functional maps. One commonly used class of computational models proposes that such maps are generated by a mechanism that projects the multiple dimensions of neuronal responses smoothly onto the two dimensions of cortex. In this issue of Neuron, Mriganka Sur and colleagues find a close match between such model predictions and measurements from ferret V1. PMID- 16039559 TI - Cadherins: actin with the cytoskeleton to form synapses. AB - Classic cadherins are calcium-dependent homophilic cell adhesion molecules that are enriched at synapses and thought to function in target recognition and adhesion at synaptic junctions. This brief review highlights evidence that cadherins and their associated catenins play a role in directing the development of pre- and postsynaptic specializations. In particular, the question of whether cadherin regulation of the actin cytoskeleton at discrete contact sites translates into the assembly of synaptic compartments will be explored. PMID- 16039560 TI - Phosphoinositide lipid second messengers: new paradigms for calcium channel modulation. AB - Neuronal Ca2+ channels are key transducers coupling excitability to cellular function. As such, they are tightly regulated by multiple G protein-signaling pathways that finely tune their activity. In addition to fast, direct G(beta)gamma modulation of Ca2+ channels, a slower Galpha(q/11)-mediated mechanism has remained enigmatic despite intensive study. Recent work suggests that membrane phosphoinositides are crucial determinants of Ca2+ channel activity. Here, we discuss their role in Ca2+ channel modulation and the leading theories that seek to elucidate the underlying molecular details of the so-called "mysterious" G(q/11)-mediated signal. PMID- 16039561 TI - Ion permeation through a voltage- sensitive gating pore in brain sodium channels having voltage sensor mutations. AB - Voltage-gated sodium channels activate in response to depolarization, but it is unknown whether the voltage-sensing arginines in their S4 segments pivot across the lipid bilayer as voltage sensor paddles or move through the protein in a gating pore. Here we report that mutation of pairs of arginine gating charges to glutamine induces cation permeation through a gating pore in domain II of the Na(V)1.2a channel. Mutation of R850 and R853 induces a K(+)-selective inward cationic current in the resting state that is blocked by activation. Remarkably, mutation of R853 and R856 causes an outward cationic current with the opposite gating polarity. These results support a model in which the IIS4 gating charges move through a narrow constriction in a gating pore in the sodium channel protein during gating. Paired substitutions of glutamine allow cation movement through the constriction when appropriately positioned by the gating movements of the S4 segment. PMID- 16039562 TI - Abeta42 is essential for parenchymal and vascular amyloid deposition in mice. AB - Considerable circumstantial evidence suggests that Abeta42 is the initiating molecule in Alzheimer's disease (AD) pathogenesis. However, the absolute requirement for Abeta42 for amyloid deposition has never been demonstrated in vivo. We have addressed this by developing transgenic models that express Abeta1 40 or Abeta1-42 in the absence of human amyloid beta protein precursor (APP) overexpression. Mice expressing high levels of Abeta1-40 do not develop overt amyloid pathology. In contrast, mice expressing lower levels of Abeta1-42 accumulate insoluble Abeta1-42 and develop compact amyloid plaques, congophilic amyloid angiopathy (CAA), and diffuse Abeta deposits. When mice expressing Abeta1 42 are crossed with mutant APP (Tg2576) mice, there is also a massive increase in amyloid deposition. These data establish that Abeta1-42 is essential for amyloid deposition in the parenchyma and also in vessels. PMID- 16039563 TI - Ptf1a, a bHLH transcriptional gene, defines GABAergic neuronal fates in cerebellum. AB - The molecular machinery governing glutamatergic-GABAergic neuronal subtype specification is unclear. Here we describe a cerebellar mutant, cerebelless, which lacks the entire cerebellar cortex in adults. The primary defect of the mutant brains was a specific inhibition of GABAergic neuron production from the cerebellar ventricular zone (VZ), resulting in secondary and complete loss of external germinal layer, pontine, and olivary nuclei during development. We identified the responsible gene, Ptf1a, whose expression was lost in the cerebellar VZ but was maintained in the pancreas in cerebelless. Lineage tracing revealed that two types of neural precursors exist in the cerebellar VZ: Ptf1a expressing and -nonexpressing precursors, which generate GABAergic and glutamatergic neurons, respectively. Introduction of Ptf1a into glutamatergic neuron precursors in the dorsal telencephalon generated GABAergic neurons with representative morphological and migratory features. Our results suggest that Ptf1a is involved in driving neural precursors to differentiate into GABAergic neurons in the cerebellum. PMID- 16039564 TI - Gliomedin mediates Schwann cell-axon interaction and the molecular assembly of the nodes of Ranvier. AB - Accumulation of Na(+) channels at the nodes of Ranvier is a prerequisite for saltatory conduction. In peripheral nerves, clustering of these channels along the axolemma is regulated by myelinating Schwann cells through a yet unknown mechanism. We report the identification of gliomedin, a glial ligand for neurofascin and NrCAM, two axonal immunoglobulin cell adhesion molecules that are associated with Na+ channels at the nodes of Ranvier. Gliomedin is expressed by myelinating Schwann cells and accumulates at the edges of each myelin segment during development, where it aligns with the forming nodes. Eliminating the expression of gliomedin by RNAi, or the addition of a soluble extracellular domain of neurofascin to myelinating cultures, which caused the redistribution of gliomedin along the internodes, abolished node formation. Furthermore, a soluble gliomedin induced nodal-like clusters of Na+ channels in the absence of Schwann cells. We propose that gliomedin provides a glial cue for the formation of peripheral nodes of Ranvier. PMID- 16039565 TI - Knockdown of cone-specific kinase GRK7 in larval zebrafish leads to impaired cone response recovery and delayed dark adaptation. AB - Phosphorylation of rhodopsin by rhodopsin kinase GRK1 is an important desensitization mechanism in scotopic vision. For cone vision GRK1 is not essential. However, cone opsin is phosphorylated following light stimulation. In cone-dominant animals as well as in humans, but not in rodents, GRK7, a cone specific homolog of GRK1, has been identified in cone outer segments. To investigate the function of GRK7 in vivo, we cloned two orthologs of grk7 in zebrafish and knocked down gene expression of grk7a in zebrafish larvae by morpholino antisense nucleotides. Photoresponse recovery in Grk7a-deficient larvae was delayed in electroretinographic measurements, and temporal contrast sensitivity was reduced, particularly under bright-light conditions. These results show that function of a cone-specific kinase is essential for cone vision in the zebrafish retina and argue that pigment bleaching and spontaneous decay alone are not sufficient for light adaptation and rapid cone response inactivation. PMID- 16039566 TI - Auditory hair cell-afferent fiber synapses are specialized to operate at their best frequencies. AB - Auditory afferent fiber activity is driven by high-fidelity information transfer from the sensory hair cell. Presynaptic specializations, posited to maintain fidelity, are investigated at synapses with characteristic frequencies of 120 Hz and 320 Hz. Morphological data indicate that high-frequency cells have more synapses and higher vesicle density near dense bodies (DBs). Tracking vesicular release via capacitance changes identified three overlapping kinetic components of release corresponding to morphologically identified vesicle pools. High frequency cells released faster; however, when normalized to release site number, low-frequency cells released faster, likely due to a greater Ca2+ load per synapse. The Ca(2+)-dependence of release was nonsaturating and independent of frequency, suggesting that release, not refilling, was rate limiting. A model of release derived from vesicle equilibration between morphologically defined pools reproduced the capacitance data, supporting a critical role in vesicle trafficking for DBs. The model suggests that presynaptic specializations enable synapses to operate most efficiently at their characteristic frequencies. PMID- 16039567 TI - Dopamine-dependent interactions between limbic and prefrontal cortical plasticity in the nucleus accumbens: disruption by cocaine sensitization. AB - The prefrontal cortex and the hippocampus exhibit converging projections to the nucleus accumbens and have functional reciprocal connections via indirect pathways. As a result, information processing between these structures is likely to be bidirectional. Using evoked potential measures, we examined the interactions of these inputs on synaptic plasticity within the accumbens. Our results show that the direction of information flow between the prefrontal cortex and limbic structures determines the synaptic plasticity that these inputs exhibit within the accumbens. Moreover, this synaptic plasticity at hippocampal and prefrontal inputs selectively involves dopamine D1 and D2 activation or inactivation, respectively. Repeated cocaine administration disrupted this synaptic plasticity at hippocampal and prefrontal cortical inputs and goal directed behavior in the spatial maze task. Thus, interactions of limbic prefrontal cortical synaptic plasticity and its dysfunction within the accumbens could underlie complex information processing deficits observed in individuals following psychostimulant administration. PMID- 16039568 TI - The coordinated mapping of visual space and response features in visual cortex. AB - Whether general principles can explain the layouts of cortical maps remains unresolved. In primary visual cortex of ferret, the relationships between the maps of visual space and response features are predicted by a "dimension reduction" model. The representation of visual space is anisotropic, with the elevation and azimuth axes having different magnification. This anisotropy is reflected in the orientation, ocular dominance, and spatial frequency domains, which are elongated such that their directions of rapid change, or high-gradient axes, are orthogonal to the high-gradient axis of the visual map. The feature maps are also strongly interdependent-their high-gradient regions avoid one another and intersect orthogonally where essential, so that overlap is minimized. Our results demonstrate a clear influence of the visual map on each feature map. In turn, the local representation of visual space is smooth, as predicted when many features are mapped within a cortical area. PMID- 16039569 TI - Do motoneurons encode the noncommutativity of ocular rotations? AB - As we look around, the orientation of our eyes depends on the order of the rotations that are carried out, a mathematical feature of rotatory motions known as noncommutativity. Theorists and experimentalists continue to debate how biological systems deal with this property when generating kinematically appropriate movements. Some believe that this is always done by neural commands to a simplified eye plant. Others have postulated that noncommutativity is implemented solely by the mechanical properties of the eyeball. Here we directly examined what the brain tells the muscles, by recording motoneuron activities as monkeys made eye movements. We found that vertical recti and superior/inferior oblique motoneurons, which drive sensory-generated torsional eye movements, do not modulate their firing rates according to the noncommutative-driven torsion during pursuit. We conclude that part of the solution for kinematically appropriate eye movements is found in the mechanical properties of the eyeball, although neural computations remain necessary and become increasingly important during head movements. PMID- 16039570 TI - Formal learning theory dissociates brain regions with different temporal integration. AB - Learning can be characterized as the extraction of reliable predictions about stimulus occurrences from past experience. In two experiments, we investigated the interval of temporal integration of previous learning trials in different brain regions using implicit and explicit Pavlovian fear conditioning with a dynamically changing reinforcement regime in an experimental setting. With formal learning theory (the Rescorla-Wagner model), temporal integration is characterized by the learning rate. Using fMRI and this theoretical framework, we are able to distinguish between learning-related brain regions that show long temporal integration (e.g., amygdala) and higher perceptual regions that integrate only over a short period of time (e.g., fusiform face area, parahippocampal place area). This approach allows for the investigation of learning-related changes in brain activation, as it can dissociate brain areas that differ with respect to their integration of past learning experiences by either computing long-term outcome predictions or instantaneous reinforcement expectancies. PMID- 16039572 TI - Divide and conquer: division of labor by B-1 B cells. AB - Capsular polysaccharides enhance bacterial virulence. In this issue of Immunity, demonstrate a division of labor between the B-1 B cell subsets: natural antibodies from B-1a cells limit infection by Streptococcus pneumoniae, whereas B 1b cells generate the anticapsule-induced response that prevents fatal infection. PMID- 16039571 TI - Prefrontal cortex activity related to abstract response strategies. AB - Many monkeys adopt abstract response strategies as they learn to map visual symbols to responses by trial and error. According to the repeat-stay strategy, if a symbol repeats from a previous, successful trial, the monkeys should stay with their most recent response choice. According to the change-shift strategy, if the symbol changes, the monkeys should shift to a different choice. We recorded the activity of prefrontal cortex neurons while monkeys chose responses according to these two strategies. Many neurons had activity selective for the strategy used. In a subsequent block of trials, the monkeys learned fixed stimulus-response mappings with the same stimuli. Some neurons had activity selective for choosing responses based on fixed mappings, others for choosing based on abstract strategies. These findings indicate that the prefrontal cortex contributes to the implementation of the abstract response strategies that monkeys use during trial-and-error learning. PMID- 16039573 TI - A newly discovered Fc receptor that explains IgG-isotype disparities in effector responses. AB - The known IgG FcRs primarily mediate inflammation by interacting with IgG1, even though IgG2 isotypes tend to be more pathogenic. A novel FcgammaR that binds IgG2 but not IgG1 has just been identified, potentially explaining differences in biological activity that are seen with various IgG isotypes. PMID- 16039574 TI - Selection and lineage specification in the thymus: commitment 4-stalled. AB - How CD4(+)CD8(+) thymocytes commit to CD4 helper versus CD8 cytotoxic lineages is a central unresolved question in developmental immunology. In this issue, show that engineering CD4 for shutoff immediately after positive selection misdirects cells to the cytotoxic lineage. The result highlights the distinction between positive selection and lineage commitment and provides new impetus for reexamining lineage models. PMID- 16039575 TI - B-1a and B-1b cells exhibit distinct developmental requirements and have unique functional roles in innate and adaptive immunity to S. pneumoniae. AB - B-1a and B-1b lymphocytes were found to exhibit specialized roles in providing immunity to Streptococcus pneumoniae and differ dramatically in their developmental requirements. Transgenic mice overexpressing CD19 (hCD19Tg) generated B-1a cells and natural antibodies that provided protection during infection, while CD19-deficient (CD19(-/-)) mice lacked B-1a cells, lacked natural antibodies, and were more susceptible to infection. By contrast, pneumococcal polysaccharide (PPS) immunization protected CD19(-/-) mice during lethal challenge, whereas hCD19Tg mice remained unprotected. This resulted from differences in the B-1b subset: the key population found to produce protective PPS-specific antibody in both wild-type and CD19(-/-) mice. Thus, CD19(-/-) mice generated B-1b cells and protective adaptive PPS-specific antibody responses, whereas hCD19Tg mice lacked B-1b cells and adaptive PPS-specific antibody responses. This reciprocal contribution of B-1a and B-1b subsets to innate and acquired immunity reveals an unexpected division of labor within the B-1 compartment that is normally balanced by their coordinated development. PMID- 16039576 TI - Cell type-specific involvement of RIG-I in antiviral response. AB - Toll-like receptors (TLRs) play an important role in antiviral response by recognizing viral components. Recently, a RNA helicase, RIG-I, was also suggested to recognize viral double-stranded RNA. However, how these molecules contribute to viral recognition in vivo is poorly understood. We show by gene targeting that RIG-I is essential for induction of type I interferons (IFNs) after infection with RNA viruses in fibroblasts and conventional dendritic cells (DCs). RIG-I induces type I IFNs by activating IRF3 via IkappaB kinase-related kinases. In contrast, plasmacytoid DCs, which produce large amounts of IFN-alpha, use the TLR system rather than RIG-I for viral detection. Taken together, RIG-I and the TLR system exert antiviral responses in a cell type-specific manner. PMID- 16039577 TI - The kinetic stability of MHC class II:peptide complexes is a key parameter that dictates immunodominance. AB - T cell priming to exogenous antigens reflects regulated antigen processing in dendritic cells, subsequent homing to lymph nodes, sustained interactions between T cells and antigen-bearing dendritic cells, and, ultimately, selective T cell activation and differentiation. In this study, we test the hypothesis that an intrinsic property of the class II:peptide complex is a key determinant that dictates the specificity of an emerging CD4 T cell response. We found that immunodominant peptides possess extremely long half-lives with class II molecules (t(1/2) > 150 hr), whereas cryptic peptides displayed half-lives of less than 10 hr. Furthermore, and most importantly, by using a peptide shuttle vector and four independent antigens, we demonstrate a direct, causative relationship between the half-life of peptide epitopes and their immunogenicity in vivo. Taken collectively, our results suggest the half-life of class II:peptide complexes is the primary parameter that dictates the ultimate hierarchy of the elicited T cell response. PMID- 16039578 TI - FcgammaRIV: a novel FcR with distinct IgG subclass specificity. AB - Mouse IgG subclasses display a hierarchy of in vivo activities, with IgG2a and IgG2b showing the greatest protective and pathogenic properties. These enhanced activities result, in part, from their ability to bind to a novel, gamma chain dependent, activating IgG Fc receptor, FcgammaRIV. FcgammaRIV maps in the 75 kb genomic interval between FcgammaRII and FcgammaRIII; its expression is restricted to myeloid lineage cells, and it binds to IgG2a and IgG2b with intermediate affinity. No binding to IgG1 or IgG3 was observed. Blocking FcgammaRIV binding to pathogenic anti-platelet antibodies is sufficient to protect mice from antibody induced thrombocytopenia. Thus, the FcgammaR system has evolved distinct activation receptors displaying selectivity for IgG subclasses, with IgG1 antibodies exclusively dependent on FcgammaRIII, whereas IgG2a and IgG2b show preferential dependence on FcgammaRIV activation. These distinct binding affinities for the IgG subclasses to FcgammaRs account for their differential protective and pathogenic activities in vivo. PMID- 16039579 TI - Oscillating CD8(+) T cell effector functions after antigen recognition in the liver. AB - When hepatitis B virus (HBV)-specific CD8(+) cytotoxic T lymphocytes (CTLs) are adoptively transferred into HBV transgenic mice, they enter the liver, recognize antigen, secrete interferon gamma (IFNgamma), inhibit viral replication, and kill their target cells, causing hepatitis. In the current study, we examined the impact of antigen recognition on the evolution of the activation phenotype, antiviral effector functions, expansion and contraction kinetics, and compartmentalization of the transferred CTLs. The results reveal that noncytolytic and cytolytic effector functions and expansion-contraction kinetics of the CTLs are regulated asynchronously and in an oscillatory manner as a consequence of antigen recognition in the liver and in association with PD-1 upregulation. We suggest that such oscillations maximize viral clearance and minimize tissue injury during HBV infection and that poor coordination of these events could lead to viral persistence and chronic liver disease. PMID- 16039580 TI - In vivo identification of novel regulators and conserved pathways of phagocytosis in A. gambiae. AB - Anopheles gambiae uses effective immune responses, including phagocytosis, to fight microbial infection. We have developed a semiquantitative phagocytosis test and used it in conjunction with dsRNA gene silencing to test the in vivo roles of 71 candidate genes in phagocytosis of Escherichia coli and Staphylococcus aureus. Here, we show that inactivation of 26 genes changes the phagocytic activity by more than 45% and that two pathways similar to those that mediate apoptotic cell removal in Caenorhabditis elegans are used in A. gambiae for phagocytosis of microorganisms. Simultaneous inactivation of the identified regulators of phagocytosis and conserved components defining each signaling pathway permitted provisional assignment of the novel regulators to one or the other pathway. Pathway inactivation enhances at least three times the ability of E. coli and S. aureus to proliferate in the mosquito. Interestingly, mosquito survival is not compromised even if both pathways are perturbed simultaneously. PMID- 16039581 TI - Modulation of coreceptor transcription during positive selection dictates lineage fate independently of TCR/coreceptor specificity. AB - For developing T cells, coreceptor choice is matched to T cell antigen receptor (TCR) MHC specificity during positive selection in the thymus, but the mechanism remains uncertain. Here, we document that TCR-mediated positive selection signals inactivate the immature CD8(III) enhancer in double positive (DP) thymocytes, explaining in part the cessation of CD8 coreceptor transcription that occurs during positive selection. More importantly, by placing CD4 protein expression under the control of CD8 transcriptional regulatory elements, we demonstrate that cessation of CD4 coreceptor transcription during positive selection results in precisely the same lineage fate as cessation of CD8 coreceptor transcription. That is, MHC-II-signaled DP thymocytes differentiated into CD8-lineage cytotoxic T cells, despite the MHC-II specificity and CD4 dependence of their TCRs. This study demonstrates that termination of coreceptor transcription during positive selection promotes CD8-lineage fate, regardless of TCR specificity or coreceptor protein identity. PMID- 16039582 TI - Probabilistic regulation in TH2 cells accounts for monoallelic expression of IL-4 and IL-13. AB - Il4 and Il13, closely linked genes, are expressed monoallelically in TH2 cells. Four different approaches (RNA FISH, cultures from Il13T-Il4/Il13-G4 mice, cultures from heterozygous Il13-Il4 double knockout mice, and a highly selected set of BABL/c*CAST/Ei clones displaying strong Il4 allelic bias) were utilized to study monoallelic expression of Il4 and coexpression of Il4 and Il13 on the same chromosome. There was a random probability for expression of one or two Il4 and one or two Il13 alleles; coexpression of cis and trans Il4 and Il13 alleles was equally probable. Histone H3 acetylation of CNS1, located in the Il13-Il4 intergenic region, was permissive for expression of IL-4 and IL-13 but did not determine the degree of their expression. Thus, monoallelism at the Il4 locus is a complex process; expression is linked to opening CNS1 but probability of expression is controlled at other sites. Based on these probabilities, individual cells randomly express Il4 and Il13 alleles. PMID- 16039583 TI - Histone modifications associated with somatic hypermutation. AB - A number of modified histones, including acetylated H3 and H4 and phosphorylated H2AX (gammaH2AX), are associated with V(D)J recombination and class switch recombination (CSR). In contrast, little is known concerning the chromatin modifications associated with somatic hypermutation (SHM) in vivo. Here, we report that several modifications--including histone acetylation and H3-lysine 4 methylation--fail to demarcate an actively hypermutating immunoglobulin (Ig) locus or to correlate spatially with SHM within Ig loci. Furthermore, no obvious association between SHM and gammaH2AX could be detected. Instead, we find that the phosphorylated form of histone H2B (H2B(Ser14P)) correlates tightly with SHM and CSR. Phosphorylation of H2B within Ig variable and switch regions requires AID and may be mediated by the histone kinase Mst1. These findings indicate that SHM and CSR trigger distinct DNA damage responses and identify a novel histone modification pattern for SHM consisting of H2B(Ser14P) in the absence of gammaH2AX. PMID- 16039584 TI - Cyclophilin B escorts the hepatitis C virus RNA polymerase: a viral achilles heel? AB - A recent report by Watashi et al. (2005) in Molecular Cell reveals a role for the host cell prolyl isomerase cyclophilin B (CyPB) in the replication of the hepatitis C viral genome, opening potential avenues for antiviral therapeutic intervention. PMID- 16039585 TI - The dynamics of chromatin remodeling at promoters. AB - The nucleosome, the structural unit of chromatin, is known to play a central role in regulating gene transcription from promoters. The last seven years have spawned a vast amount of data on the enzymes that remodel and modify nucleosomes and the rules governing how transcription factors interact with the epigenetic code on histones. Yet despite this effort, there has yet to emerge a unifying mechanism by which nucleosomes are remodeled during gene regulation. Recent advances have allowed nucleosome dynamics on promoters to be studied in real time, dramatically changing how we think about gene regulation on chromatin templates. PMID- 16039586 TI - Erk associates with and primes GSK-3beta for its inactivation resulting in upregulation of beta-catenin. AB - Beta-catenin is upregulated in many human cancers and considered to be an oncogene. Hepatocellular carcinoma (HCC) is one of the most prevalent human malignancies, and individuals who are chronic hepatitis B virus (HBV) carriers have a greater than 100-fold increased relative risk of developing HCC. Here we report a mechanism by which HBV-X protein (HBX) upregulates beta-catenin. Erk, which is activated by HBX, associates with GSK-3beta through a docking motif ((291)FKFP) of GSK-3beta and phosphorylates GSK-3beta at the (43)Thr residue, which primes GSK-3beta for its subsequent phosphorylation at Ser9 by p90RSK, resulting in inactivation of GSK-3beta and upregulation of beta-catenin. This pathway is a general signal, as it was also observed in cell lines in which Erk primed inactivation of GSK-3beta was regulated by IGF-1, TGF-beta, and receptor tyrosine kinase HER2, and is further supported by immunohistochemical staining in different human tumors, including cancers of the liver, breast, kidney, and stomach. PMID- 16039587 TI - Involvement of heme regulatory motif in heme-mediated ubiquitination and degradation of IRP2. AB - Iron regulatory protein 2 (IRP2), a regulator of iron metabolism, is modulated by ubiquitination and degradation. We have shown that IRP2 degradation is triggered by heme-mediated oxidation. We report here that not only Cys201, an invariant residue in the heme regulatory motif (HRM), but also His204 is critical for IRP2 degradation. Spectroscopic studies revealed that Cys201 binds ferric heme, whereas His204 is a ferrous heme binding site, indicating the involvement of these residues in sensing the redox state of the heme iron and in generating the oxidative modification. Moreover, the HRM in IRP2 has been suggested to play a critical role in its recognition by the HOIL-1 ubiquitin ligase. Although HRMs are known to sense heme concentration by simply binding to heme, the HRM in IRP2 specifically contributes to its oxidative modification, its recognition by the ligase, and its sensing of iron concentration after iron is integrated into heme. PMID- 16039588 TI - More than one glycan is needed for ER glucosidase II to allow entry of glycoproteins into the calnexin/calreticulin cycle. AB - Nascent and newly synthesized glycoproteins enter the calnexin (Cnx)/calreticulin (Crt) cycle when two out of three glucoses in the core N-linked glycans have been trimmed sequentially by endoplasmic reticulum (ER) glucosidases I (GI) and II (GII). By analyzing arrested glycopeptides in microsomes, we found that GI removed the outermost glucose immediately after glycan addition. However, although GII associated with singly glycosylated nascent chains, trimming of the second glucose only occurred efficiently when a second glycan was present in the chain. Consistent with a requirement for multiple glycans to activate GII, pancreatic RNase in live cells needed more than one glycan to enter the Cnx/Crt cycle. Thus, whereas GI trimming occurs as an automatic extension of glycosylation, trimming by GII is a regulated process. By adjusting the number and location of glycans, glycoproteins can instruct the cell to engage them in an individually determined folding and quality control pathway. PMID- 16039589 TI - Involvement of a mitochondrial phosphatase in the regulation of ATP production and insulin secretion in pancreatic beta cells. AB - Reversible phosphorylation is the cell's most prevalent form of posttranslational modification, yet its role in the regulation of mitochondrial functions is poorly understood. We have discovered that a member of the dual-specific protein tyrosine phosphatase (DS-PTP) family, PTPMT1 (PTP localized to the Mitochondrion 1) resides nearly exclusively in mitochondria. PTPMT1 is targeted to the mitochondrion by an N-terminal signal sequence and is found anchored to the matrix face of the inner membrane. Knockdown of PTPMT1 expression in the pancreatic insulinoma cell line INS-1 832/13 alters the mitochondrial phosphoprotein profile and markedly enhances both ATP production and insulin secretion. These data define PTPMT1 as a potential drug target for the treatment of type II diabetes and strengthen the notion that mitochondria are an underappreciated site of signaling by reversible phosphorylation. PMID- 16039590 TI - Kin4 kinase delays mitotic exit in response to spindle alignment defects. AB - For many polarized cells, it is critical that the mitotic spindle becomes positioned relative to the polarity axis. This is especially important in yeast, where the site of cytokinesis is predetermined. The spindle position checkpoint (SPOC) therefore delays mitotic exit of cells with a mispositioned spindle. One component of the SPOC is the Bub2-Bfa1 complex, an inhibitor of the mitotic exit network (MEN). Here, we show that the Kin4 kinase is a component of the SPOC and as such is essential to delay cell cycle progression of cells with a misaligned spindle. When spindles are correctly oriented, Kin4 and Bub2-Bfa1 are asymmetrically localized to opposite spindle pole bodies (SPBs). Bub2-Bfa1 then becomes inhibited by Cdc5 polo kinase with anaphase onset, a prerequisite for mitotic exit. In response to spindle misalignment, Kin4 and Bub2-Bfa1 are brought together at both SPBs. Kin4 now maintains Bub2-Bfa1 activity by counteracting Cdc5, thereby inhibiting mitotic exit. PMID- 16039591 TI - The protein kinase Kin4 inhibits exit from mitosis in response to spindle position defects. AB - Accurate nuclear position is essential for each daughter cell to receive one DNA complement. In budding yeast, a surveillance mechanism known as the spindle position checkpoint ensures that exit from mitosis only occurs when the anaphase nucleus is positioned along the mother-bud axis. We identified the protein kinase Kin4 as a component of the spindle position checkpoint. KIN4 prevents exit from mitosis in cells with mispositioned nuclei by inhibiting the mitotic exit network (MEN), a GTPase signaling cascade that promotes exit from mitosis. Kin4 is active in cells with mispositioned nuclei and predominantly localizes to mother cells, where it is ideally situated to inhibit MEN signaling at spindle pole bodies (SPBs) when anaphase spindle elongation occurs within the mother cell. PMID- 16039592 TI - molecular mechanism of lysidine synthesis that determines tRNA identity and codon recognition. AB - Lysidine (2-lysyl cytidine) is a lysine-containing cytidine derivative commonly found at the wobble position of bacterial AUA codon-specific tRNA(Ile). This modification determines both codon and amino acid specificities of tRNA(Ile). We previously identified tRNA(Ile)-lysidine synthetase (tilS) that synthesizes lysidine, for which it utilizes ATP and lysine as substrates. Here, we show that lysidine synthesis consists of two consecutive reactions that involve an adenylated tRNA intermediate. A mutation study revealed that Escherichia coli TilS discriminates tRNA(Ile) from the structurally similar tRNA(Met) having the same anticodon loop by recognizing the anticodon loop, the anticodon stem, and the acceptor stem. TilS was shown to bind to the anticodon region and 3' side of the acceptor stem, which cover the recognition sites. These findings reveal a dedicated mechanism embedded in tRNA(Ile) that controls its recognition and discrimination by TilS, and indicate the significance of this enzyme in the proper deciphering of genetic information. PMID- 16039593 TI - RNA polymerase modulators and DNA repair activities resolve conflicts between DNA replication and transcription. AB - Organisms rely on close interplay between DNA replication, recombination, and repair to secure transmission of the genome. In rapidly dividing cells, there is also great pressure for transcription, which may induce conflict with replication. We investigated the potential for conflict in bacterial cells, where there is no temporal separation of these processes. Eliminating the stringent response regulators ppGpp and DksA or the GreA and Mfd proteins, which revive or dislodge stalled transcription complexes, and especially combinations of these factors, is shown to severely reduce viability when DNA repair is also compromised. Both ppGpp and certain RNA polymerase (RNAP) mutations reduce accumulation of backed-up arrays of stalled transcription complexes. We propose these arrays are formidable obstacles to replication that are normally kept in check in wild-type cells by ppGpp, DksA, GreA, and Mfd. When arrays do obstruct replication, the consequences are resolved by one of the many pathways available to rescue stalled forks. PMID- 16039594 TI - Crystal structure of bacteriophage lambda cII and its DNA complex. AB - The tetrameric cII protein from bacteriophage lambda activates transcription from the phage promoters P(RE), P(I), and P(AQ) by binding to two direct repeats that flank the promoter -35 element. Here, we present the X-ray crystal structure of cII alone (2.8 A resolution) and in complex with its DNA operator from P(RE) (1.7 A resolution). The structures provide a basis for modeling of the activation complex with the RNA polymerase holoenzyme, and point to the key role for the RNA polymerase alpha subunit C-terminal domain (alphaCTD) in cII-dependent activation, which forms a bridge of protein/protein interactions between cII and the RNA polymerase sigma subunit. The model makes specific predictions for protein/protein interactions between cII and alphaCTD, and between alphaCTD and sigma, which are supported by previous genetic studies. PMID- 16039595 TI - Histone H2B ubiquitylation controls processive methylation but not monomethylation by Dot1 and Set1. AB - Methylation is a relatively stable histone modification, yet regulation of the transition between mono-, di-, and trimethylation of lysine (K) residues may control dynamic processes such as transcription and DNA repair. Identifying factors that regulate the ability of methyltransferases to perform successive rounds of methylation on the same lysine residue is important for understanding the functions of histone methylation. Previous reports have indicated that ubiquitylation of histone H2B K123 is required for methylation of lysines 4 and 79 of histone H3 by the methyltransferases Set1 and Dot1, respectively. In contrast, by using chromatin immunoprecipitation and mass spectrometry, we find that ubiquitylation of H2B-K123 is dispensable for monomethylation of H3-K4 and H3-K79 but is required for the transition from monomethylation to subsequent methylation states. Dot1 binding to chromatin occurs normally in the absence of H2B-K123 ubiquitylation, suggesting that ubiquitylation does not regulate enzyme recruitment but does regulate the processive activity of the histone methyltransferase. PMID- 16039596 TI - Histones are incorporated in trans during reassembly of the yeast PHO5 promoter. AB - In yeast, remodeling of PHO5 promoter chromatin upon activation is accompanied by transient hyperacetylation and subsequent eviction of histones from the promoter in trans. In the course of rerepression, nucleosomes have to be reassembled on the promoter. We have analyzed where the histones for reassembly of the inactive promoter chromatin come from. The use of a strain with two differently tagged and differently regulated versions of histone H3 allowed us to discriminate between histones originating from the chromatin fraction and histones arising from the soluble histone pool. In this way, we show that the incorporated histones originate from a source in trans. Promoter closure occurs very rapidly, and the histone chaperones Asf1 and Hir1 as well as the SWI/SNF nucleosome remodeling complex appear to be important for rapid reassembly of nucleosomes at the PHO5 promoter. PMID- 16039597 TI - Fatalities in Swedish skydiving. AB - Exact risk patterns in skydiving fatalities are not well known, but incomplete world injury data indicate that many are preventable. A comprehensive national material for Sweden of 37 skydiving fatalities 1964-2003 were reviewed to identify risk factors. In relation to jump volume, the period 1994-2003 had a fatality rate 11 times lower than 1964-1973. Student skydivers had the highest risk of fatal outcome, often caused by instability in freefall leading to unstable parachute activation with subsequent line entanglement, or parachute activation failure. Unintentional water landings also contributed to student fatality, with life jacket malfunctions, neglect to use life jackets, and automatic reserve parachute activation devices activated by water as aggravating factors. One-third of all fatalities had an inflated and operational parachute at some point prior to injury. A drastic worldwide increase in fatal landing incidents with fast wing parachutes during the 1990s did not occur in Sweden. Every fourth fatality caused by rapid deceleration against ground or water survived impact and died during transports or in hospitals. Rescue units and health care providers can improve management of skydiving incidents from knowledge about the incident and injury mechanisms we have described, and the skydiving community can target risk factors in preventive safety work. PMID- 16039598 TI - Four-color multiplex reverse transcription polymerase chain reaction--overcoming its limitations. AB - Quantitative reverse transcription polymerase chain reaction (qRT-PCR) conducted in real time is a powerful tool for measuring messenger RNA (mRNA) levels in biological samples. Multiplex PCR is defined as the simultaneous amplification of two or more DNA (cDNA) targets in a single reaction vessel and may be carried out only using uniquely labeled probes for each target. Up to four genes can be detected in a multiplex 5' nuclease assay when using the appropriate instrument and the right combination of fluorophores. One of the more important advantages of multiplexing is a reduced sample requirement, which is especially important when sample material is scarce. Additional benefits are saving time on reaction setup and lower cost compared to singleplex reactions. Although multiplexing has several advantages over singleplex qRT-PCR, limited work has been done to show its feasibility. Few publications on four-color multiplex qRT-PCR have been reported, and to our knowledge no work has been done to explore the assay's limitations. In this paper, we report the first in-depth analysis of a four-gene multiplex qRT-PCR. To achieve a better understanding of the potential limitations of the qRT-PCR assay, we used in vitro transcribed RNA derived from four human genes. To emulate gene expression experiments, we developed a model system in which the in vitro transcripts were spiked with plant total RNA. This model allowed us to develop an artificial system closely resembling differential gene expression levels varying up to a million fold. We identified a single "universal" reaction condition that enabled optimal amplification in real time of up to four genes over a wide range of template concentrations. This study shows that multiplexing is a feasible approach applicable to most qRT-PCR assays performed with total RNA, independent of the expression levels of the genes under scrutiny. PMID- 16039599 TI - Amperometric biosensor for choline based on layer-by-layer assembled functionalized carbon nanotube and polyaniline multilayer film. AB - Conducting polymer film was prepared by electrochemical polymerization of aniline. Multiwalled carbon nanotubes (MWNTs) were treated with a mixture of concentrated sulfuric and nitric acid to introduce carboxylic acid groups to the nanotubes. By using the layer-by-layer method, homogeneous and stable MWNTs and polyaniline (PANI) multilayer films were alternately assembled on glassy carbon (GC) electrodes. Conducting polymer of PANI had three main functions: (i) excellent antiinterference ability, (ii) protection ability in favor of increasing the amount of the MWNTs immobilized on GC electrodes, and (iii) superior transducing ability. The protection effect of PANI film and the electrostatic interaction between positively charged PANI and negatively charged MWNTs both attributed to immobilizing abundant MWNTs stably, thereby enhancing the catalytic activity. The layer-by-layer assembled MWNTs and PANI-modified GC electrodes offered a significant decrease in the overvoltage for the H2O2 and were shown to be excellent amperometric sensors for H2O2 from +0.2V over a wide range of concentrations. As an application example, by linking choline oxidase (CHOD), an amplified biosensor toward choline was prepared. The choline biosensor exhibited a linear response range of 1x10(-6) to 2x10(-3) M with a correlation coefficient of 0.997, and the response time and detection limit (S/N=3) were determined to be 3 s and 0.3 microM, respectively. The antiinterference biosensor displays a rapid response and an expanded linear response range as well as excellent reproducibility and stability. PMID- 16039600 TI - A continuous fluorimetric assay for protoporphyrinogen oxidase by monitoring porphyrin accumulation. AB - A continuous spectrofluorimetric assay for protoporphyrinogen oxidase (PPO, EC 1.3.3.4) activity has been developed using a 96-well plate reader. Protoporphyrinogen IX, the tetrapyrrole substrate, is a colorless nonfluorescent compound. The evolution of the fluorescent tetrapyrrole product, protoporphyrin IX, was detected using a fluorescence plate reader. The apparent Km (Kapp) values for protoporphyrinogen IX were measured as 3.8+/-0.3, 3.6+/-0.5, and 1.0+/-0.1 microM for the enzymes from human, Myxococcus xanthus, and Aquifex aeolicus, respectively. The Ki for acifluorfen, a diphenylether herbicide, was measured as 0.53 microM for the human enzyme. Also, the specific activity of mouse liver mitochondrial PPO was measured as 0.043 nmol h-1/mg mitochondria, demonstrating that this technique is useful for monitoring low-enzyme activities. This method can be used to accurately measure activities as low as 0.5 nM min-1, representing a 50-fold increase in sensitivity over the currently used discontinuous assay. Furthermore, this continuous assay may be used to monitor up to 96 samples simultaneously. These obvious advantages over the discontinuous assay will be of importance for both the kinetic characterization of recombinant PPOs and the detection of low concentrations of this enzyme in biological samples. PMID- 16039601 TI - Comparison of work rates, energy expenditure, and perceived exertion during a 1-h vacuuming task with a backpack vacuum cleaner and an upright vacuum cleaner. AB - The purpose of this study was to evaluate two types of industrial vacuum cleaners, in terms of cleaning rates, energy expenditure, and perceived exertion. Twelve industrial cleaners (six males and six females, age 28-39 yr) performed two 1-h vacuuming tasks with an upright vacuum cleaner (UVC) and a backpack vacuum cleaner (BPVC). Measures for oxygen uptake (VO2) and ratings of perceived exertion (RPE) were collected continuously during the 1-h vacuuming tasks. Cleaning rates for the UVC and BPVC were 7.23 and 14.98 m2min(-1), respectively. On a separate day subjects performed a maximal treadmill exercise test to determine their maximal aerobic capacity (peak VO2). Average absolute energy costs (in Metabolic equivalents), relative energy costs of the vacuum task compared to the subjects' maximal aerobic capacity (% peak VO2), and RPE responses for the 1-h vacuuming tasks were similar between vacuum cleaners, but % peak VO2 and RPE values differed between genders. These results indicate that the BPVC was more efficient than the UVC. With the BPVC, experienced workers vacuumed at a cleaning rate 2.07 times greater than the UVC and had similar levels of energy expenditure and perceived effort, compared to the slower cleaning rate with the UVC. PMID- 16039602 TI - The environment influences whether high-fat foods are associated with palatable or with unhealthy. AB - This study investigated whether relatively automatic evaluations of food differ between situations and between obese people and lean controls. These evaluations were assessed in the affective priming paradigm (APP) -- a response latency based measure for associations. In Experiment 1, we either focused participants (33 obese and 26 lean controls) on the palatability (restaurant condition) or on the healthiness (health condition) of food, prior to the APP. Independent of weight status, relatively automatic evaluations of food were based on palatability in the restaurant condition, and on health in the health condition. So, the current focus of attention can shape the way foods are evaluated relatively automatically. In Experiment 2, craving was induced in participants (27 obese and 29 lean controls). Unexpectedly, the craving induction did not achieve its goal of focusing on the palatability of food in general, but just for low-fat foods, possibly because of the health-emphasizing environment -- a hospital. Interestingly, obese people showed a stronger palatability priming effect with increasing levels of initial craving. For normal weight controls the effect was in the same direction, but missed significance. In our environment, palatability of food may be too salient, and health may not be salient enough, influencing automatic food-evaluations. PMID- 16039603 TI - A controlled evaluation of acceptance and commitment therapy plus habit reversal for trichotillomania. AB - This randomized trial compared a combined Acceptance and Commitment Therapy/Habit Reversal Training (ACT/HRT) to a waitlist control in the treatment of adults with trichotillomania (TTM). Twenty-five participants (12 treatment and 13 waitlist) completed the trial. Results demonstrated a significant reduction in hair pulling severity, impairment ratings, and hairs pulled, along with significant reductions in experiential avoidance and both anxiety and depressive symptoms in the ACT/HRT group compared to the waitlist control. Reductions generally were maintained at a 3-month follow-up. Decreases in experiential avoidance and greater treatment compliance were significantly correlated with reductions in TTM severity, implying that targeting experiential avoidance may be useful in the treatment of TTM. Other implications and suggestions for future research are noted. PMID- 16039604 TI - Cognitive therapy for irritable bowel syndrome is associated with reduced limbic activity, GI symptoms, and anxiety. AB - This study sought to identify brain regions that underlie symptom changes in severely affected IBS patients undergoing cognitive therapy (CT). Five healthy controls and 6 Rome II diagnosed IBS patients underwent psychological testing followed by rectal balloon distention while brain neural activity was measured with O-15 water positron emission tomography (PET) before and after a brief regimen of CT. Pre-treatment resting state scans, without distention, were compared to post-treatment scans using statistical parametric mapping (SPM). Neural activity in the parahippocampal gyrus and inferior portion of the right cortex cingulate were reduced in the post-treatment scan, compared to pre treatment (x, y, z coordinates in MNI standard space were -30, -12, -30, P=0.017; 6, 34, -8, P=0.023, respectively). Blood flow values at these two sites in the controls were intermediate between those in the pre- and post-treatment IBS patients. Limbic activity changes were accompanied by significant improvements in GI symptoms (e.g., pain, bowel dysfunction) and psychological functioning (e.g., anxiety, worry). The left pons (-2, -26, -28, P=0.04) showed decreased neural activity which was correlated with post-treatment anxiety scores. Changes in neural activity of cortical-limbic regions that subserve hypervigilance and emotion regulation may represent biologically oriented change mechanisms that mediate symptom improvement of CT for IBS. PMID- 16039605 TI - The effect of insulin deficiency on tau and neurofilament in the insulin knockout mouse. AB - Complications of diabetes mellitus within the nervous system are peripheral and central neuropathy. In peripheral neuropathy, defects in neurofilament and microtubules have been demonstrated. In this study, we examined the effects of insulin deficiency within the brain in insulin knockout mice (I-/-). The I-/- exhibited hyperphosphorylation of tau, at threonine 231, and neurofilament. In addition, we showed hyperphosphorylation of c-Jun N-terminal kinase (JNK) and glycogen synthase kinase 3 beta (GSK-3 beta) at serine 9. Extracellular signal regulated kinase 1 (ERK 1) showed decrease in phosphorylation, whereas ERK 2 showed no changes. Ultrastructural examination demonstrated swollen mitochondria, endoplasmic reticulum, and Golgi apparatus, and dispersion of the nuclear chromatin. Microtubules showed decrease in the number of intermicrotubule bridges and neurofilament presented as bunches. Thus, lack of insulin brain stimulation induces JNK hyperphosphorylation followed by hyperphosphorylation of tau and neurofilament, and ultrastructural cellular damage, that over time may induce decrease in cognition and learning disabilities. PMID- 16039606 TI - Protease-activated quantum dot probes. AB - We have developed a novel nanoparticulate luminescent probe with inherent signal amplification upon interaction with a targeted proteolytic enzyme. This construct may be useful for imaging in cancer detection and diagnosis. In this system, quantum dots (QDs) are bound to gold nanoparticles (AuNPs) via a proteolytically degradable peptide sequence to non-radiatively suppress luminescence. A 71% reduction in luminescence was achieved with conjugation of AuNPs to QDs. Release of AuNPs by peptide cleavage restores radiative QD photoluminescence. Initial studies observed a 52% rise in luminescence over 47 h of exposure to 0.2 mg/mL collagenase. These probes can be customized for targeted degradation simply by changing the sequence of the peptide linker. PMID- 16039607 TI - The cyclin-dependent kinase 11 interacts with NOT2. AB - The caspase-processed cyclin-dependent kinase 11 (formerly known as PITSLRE) is implicated in apoptotic signaling. However, the mechanism of apoptotic signal transduction through CDK11(p46) is still unclear. We used a yeast two-hybrid screening strategy and identified NOT2 as an interacting partner of caspase processed C-terminal kinase domain of CDK11 (CDK11(p46)). We demonstrate that CDK11(p46) directly interacts with NOT2 in vitro and in human cells. The NOT domain in the C-terminal part of NOT2 is responsible for the association between CDK11(p46) and NOT2. Both NOT2 and CDK11(p46) predominantly co-localized in the nucleus. Furthermore, we show that overexpression of NOT2 reduces luciferase mRNA and induces apoptosis. However, NOT2 is not phosphorylated by CDK11(p46). These findings suggest that CDK11 may contribute to apoptosis by regulating the activity of NOT2 independent of its kinase activity. PMID- 16039608 TI - A truncated FAK lacking the FERM domain displays high catalytic activity but retains responsiveness to adhesion-mediated signals. AB - In order to determine the role of the FERM domain in the regulation of FAK phosphorylation at Tyr-397, the major autophosphorylation site, we generated a truncated FAK lacking a region of the N-terminus corresponding to amino acids 1 384 (FAKDelta384). FAKDelta384 showed a striking increase in phosphorylation, as compared with wild type FAK, in lysates of either HEK 293 or FAK-/- cells. Interestingly, the truncated form of FAK lacking the N-terminal domain retains responsiveness to integrin-mediated signals, as judged by its dephosphorylation by holding cells in suspension and by the recovery of the phosphorylation when replating the cells on fibronectin. We propose a model in which removal of FERM mediated auto-inhibition is important to increase FAK catalytic activity but the translocation and clustering of this enzyme at the focal adhesions is required for maximal phosphorylation at Tyr-397. PMID- 16039609 TI - Functional reconstitution of a crenarchaeal splicing endonuclease in vitro. AB - Sulfolobus tokodaii strain 7 is one of Crenarchaea whose entire genome has been sequenced. The genome sequence revealed that it possesses two open reading frames (ORFs) that are homologous to EndA, a protein responsible for splicing endonuclease activity in Archaea. Interestingly, one of the two ORFs lacks a putative catalytic amino acid residue for the nuclease activity. To investigate their functions, the two ORF products were individually expressed in Escherichia coli, partially purified, and tested for their nuclease activities in vitro. Using in vitro transcribed tRNA precursor as a substrate, we found that the two ORF products are concurrently required to cleave exon-intron junctions. Our finding implies that the splicing endonuclease for the organism is a multi subunit complex composed of the two endA gene products. PMID- 16039610 TI - Inhibition by adiponectin of IL-8 production by human macrophages upon coculturing with late apoptotic cells. AB - Adiponectin, an adipocyte-derived hormone, reportedly suppresses the production of TNF-alpha and IL-6 by LPS-stimulated human or porcine macrophages, and the phagocytosis of microbeads by human macrophages. In this study, we used a high molecular weight form of adiponectin purified from human plasma to examine its effects on the phagocytosis of late apoptotic cells by human macrophages and the subsequent IL-8 production. Adiponectin suppressed both the phagocytosis of apoptotic cells and the IL-8 production. In contrast, adiponectin augmented both the phagocytosis of apoptotic cells and the IL-8 production in the presence of LPS. These results suggest that adiponectin is not an anti-inflammatory hormone but rather a dual modulator of innate responses. PMID- 16039611 TI - Characterisation of acyltransferases from Synechocystis sp. PCC6803. AB - As phylogenetic ancestors of plant chloroplasts cyanobacteria resemble plastids with respect to lipid and fatty acid composition. These membrane lipids show the typical prokaryotic fatty acid pattern in which the sn-2 position is exclusively esterified by C(16) acyl groups. In the course of de novo glycerolipid biosynthesis this prokaryotic fatty acid pattern is established by the sequential acylation of glycerol-3-phosphate with acyl-ACPs by the activity of different acyltransferases. In silico approaches allowed the identification of putative Synechocystis acyltransferases involved in glycerolipid metabolism. Functional expression studies in Escherichia coli showed that sll1848 codes for a lysophosphatidic acid acyltransferase with a high specificity for 16:0-ACP, whereas slr2060 encodes a lysophospholipid acyltransferase, with a broad acyl-ACP specificity but a strong preference for lysophosphatidyglycerol especially its sn 2 acyl isomer as acyl-acceptor. The generation and analysis of the corresponding Synechocystis knockout mutants revealed that lysophosphatidic acid acyltransferase unlike the lysophospholipid acyltransferase is essential for the vital functions of the cells. PMID- 16039612 TI - Plasmid-borne catabolism of methyl parathion and p-nitrophenol in Pseudomonas sp. strain WBC-3. AB - Pseudomonas sp. strain WBC-3 utilises methyl parathion (MP) or p-nitrophenol (PNP) as the sole source of carbon, nitrogen, and energy. A plasmid designated pZWL0 of approximately 70 kb in this strain was found to be responsible for MP and PNP degradation. This was based on the fact that the plasmid-cured strains showed PNP- MP- phenotype and the PNP+ MP+ phenotype could be conjugally transferred. We have also cloned a 3.4-kb HindIII fragment which exhibited methyl parathion hydrolase activity, which revealed a methyl parathion hydrolase (mph) gene whose DNA sequence is 99.5% identical to the recently identified mpd gene from Plesiomonas sp. M6 [C. Zhongli, L. Shunpeng, F. Guoping, Isolation of methyl parathion-degrading strain M6 and cloning of the methyl parathion hydrolase gene, Appl. Environ. Microbiol. 67 (2001) 4922-4925]. The mph gene was functionally expressed in Escherichia coli and the relative activities of the enzyme against different substrates were determined. The sequence alignment and phylogenetic analysis suggested that MPH and MPD evolved independently from other well-studied organophosphate hydrolases and may be originated from class B beta-lactamase family. Subsequently obtained a 6.5-kb KpnI and BamHI fragment containing the above HindIII fragment revealed that the mph gene was physically located in a typical transposon. PMID- 16039613 TI - Dynamic regulation of molecular chaperone gene expression in polyglutamine disease. AB - Expanded polyglutamine disease proteins cause adult-onset progressive neurodegeneration. Constitutive overexpression of the Hsp70 molecular chaperone is capable of suppressing polyglutamine neurodegeneration. We showed that endogenous Hsp70 expression was induced, at both transcriptional and translational levels, in Drosophila models of polyglutamine disease. Soon after the endogenous Hsp70 induction reached a maximum level at larval stage, its expression declined progressively with age. We further showed that cellular heat shock response remained intact in aged flies, indicating the decline of Hsp70 levels observed in polyglutamine-expressing flies is not due to normal ageing. In contrast to the well-documented polyglutamine suppression caused by constitutive Hsp70 overexpression, no suppression of degeneration was observed when inducible copies of hsp70 transgenes were instead coexpressed. This supports a transcriptional dysregulation of endogenous hsp70 gene induction in polyglutamine flies. Altogether, we propose that transcriptional malfunctioning of molecular chaperone gene expression contributes to the late-onset and progressive nature of polyglutamine toxicity. PMID- 16039614 TI - Activation of ERK during DNA damage-induced apoptosis involves protein kinase Cdelta. AB - We have previously shown that protein kinase C (PKC) acts upstream of caspases to regulate cisplatin-induced apoptosis. Since extracellular signal-regulated kinases (ERKs) have also been implicated in DNA damage-induced apoptosis, we have examined if ERK signaling pathway acts downstream of PKC in the regulation of cisplatin-induced apoptosis. PKC activator PDBu induced ERK1/2 phosphorylation which was inhibited by general PKC inhibitor bisindolylmaleimide and Go 6983 as well as the MEK inhibitor U0126 but not by the PKCdelta inhibitor rottlerin. Cisplatin caused a concentration-dependent activation of ERK1/2 in HeLa cells. The level of ERK2 was decreased in HeLa cells that acquired resistance to cisplatin (HeLa/CP). The MEK inhibitor U0126 inhibited cisplatin-induced ERK activation and attenuated cisplatin-induced cell death. Inhibition of PKCdelta by rottlerin or depletion of PKCdelta by siRNA inhibited cisplatin-induced ERK activation. These results suggest that cisplatin-induced DNA damage results in activation of ERK1/2 via PKCdelta. PMID- 16039615 TI - VEGF induces proliferation, migration, and TGF-beta1 expression in mouse glomerular endothelial cells via mitogen-activated protein kinase and phosphatidylinositol 3-kinase. AB - The role of glomerular endothelial cells in kidney fibrosis remains incompletely understood. While endothelia are indispensable for repair of acute damage, they can produce extracellular matrix proteins and profibrogenic cytokines that promote fibrogenesis. We used a murine cell line with all features of glomerular endothelial cells (glEND.2), which dissected the effects of vascular endothelial growth factor (VEGF) on cell migration, proliferation, and profibrogenic cytokine production. VEGF dose-dependently induced glEND.2 cell migration and proliferation, accompanied by up-regulation of VEGFR-2 phosphorylation and mRNA expression. VEGF induced a profibrogenic gene expression profile, including up regulation of TGF-beta1 mRNA, enhanced TGF-beta1 secretion, and bioactivity. VEGF induced endothelial cell migration and TGF-beta1 induction were mediated by the phosphatidyl-inositol-3 kinase pathway, while proliferation was dependent on the Erk1/2 MAP kinase pathway. This suggests that differential modulation of glomerular angiogenesis by selective inhibition of the two identified VEGF induced signaling pathways could be a therapeutic approach to treat kidney fibrosis. PMID- 16039616 TI - Targeted correction of a chromosomal point mutation by modified single-stranded oligonucleotides in a GFP recovery system. AB - Synthetic oligonucleotides had been employed in DNA repair and promised great potentials in gene therapy. To test the ability of single-stranded oligonucleotide (SSO)-mediated gene repair within a chromosomal site in human cells, a HeLa cell line stably integrated with mutant enhanced green fluorescence protein gene (mEGFP) in the genome was established. Transfection with specific SSOs successfully repaired the mEGFP gene and resulted in the expression of functional fluorescence proteins, which could be detected by fluorescence microscopy and FACS assay. Western blot showed that EGFP was only present in the cells transfected with correction SSOs rather than the control SSOs. Furthermore, DNA sequencing confirmed that phenotype change resulted from the designated nucleotide correction at the target site. Using this reporter system, we determined the optimal structure of SSO by investigating the effect of length, modifications, and polarities of SSOs as well as the positions of the mismatch forming nucleotide on the efficiency of SSO-mediated gene repair. Interestingly, we found that SSOs with mismatch-forming nucleotide positioned at different positions have varying potencies that homology at the 5'-end of SSOs was more crucial for the SSO's activity. These results provided guidance for designing effective SSOs as tools for treating monogenic inherited diseases. PMID- 16039617 TI - Limb chondrogenesis is compromised in the versican deficient hdf mouse. AB - It has been suggested that the matrix proteoglycan, versican, may perform a functional role during early events of limb skeletogenesis largely by virtue of its spatiotemporal expression pattern in precartilage mesenchymal aggregations. The versican-deficient hdf transgenic mouse has provided the first model to explore the implications of a null mature versican on limb chondrogenesis. Due to lethality of hdf homozygous embryos prior to limb cartilage differentiation, high density micromass cultures were employed to compare the chondrogenic capacity of hdf mutant limb mesenchyme to that of wild-type. In homozygous hdf mesenchyme, aggregation was severely compromised and neither cartilage-characteristic Type II collagen nor alcian blue positive foci were detected during a 6-day period of culture. Three-dimensional culture of hdf mutant mesenchyme, however, showed that in a permissive environment mutant cells also expressed Type II collagen. Results strongly suggest that mature versican proteoglycan is essential for precartilage aggregation and subsequent cartilage differentiation. PMID- 16039618 TI - Cognitive therapy does not prevent a response to tryptophan depletion in patients also treated with antidepressants. AB - BACKGROUND: Acute tryptophan depletion (ATD) induces depressive symptoms in remitted depressed patients treated with serotonergic medications, but not in patients treated with noradrenergic medications or electroconvulsive therapy. A recent study suggests that cognitive therapy (CT) protects against the effects of ATD, but the evidence is questionable. The present study compared the effect of ATD in patients who were treated with antidepressant medication and CT (n = 17) versus antidepressant medication alone (n = 23) during their latest episode. METHODS: Forty remitted depressed patients underwent high-dose and low-dose ATD in a randomized double-blind crossover design. RESULTS: There were no differences in response to ATD between treatment groups. This applied to groups defined by lifetime and by recent CT experience. CONCLUSIONS: Cognitive therapy does not protect against the effects of rapidly lowered plasma tryptophan levels in remitted depressed patients who are also treated with antidepressant medication. PMID- 16039619 TI - The relationship of structural alterations to cognitive deficits in schizophrenia: a voxel-based morphometry study. AB - BACKGROUND: Region of interest studies have identified a number of structure cognition associations in schizophrenia and revealed alterations in structure cognition relationship in this population. METHODS: We examined the relationship of structural brain alterations, identified using voxel-based morphometry, to cognitive deficits in 45 schizophrenia patients relative to 43 healthy control subjects and tested the hypothesis that structure-cognition relationship is altered in schizophrenia. RESULTS: Patients had smaller total brain, gray matter, and white matter volumes. Regional alterations were left-hemisphere specific, including: gray matter reduction of inferior frontal, lingual, and anterior superior temporal gyri; white matter reduction of posterior and occipital lobes; and gray matter increase of the putamen and the precuneus. Smaller whole brain and gray matter volumes were associated with lower premorbid intelligence quotient (IQ) and poorer performance on IQ-dependent cognitive measures in patients and to a similar extent in control subjects. Larger precuneus was associated with better immediate verbal memory in patients, whereas verbal and nonverbal memory were positively associated with inferior frontal gyrus volume in control subjects. Smaller occipital white matter volume was associated with slower information processing speed in patients but not in control subjects. CONCLUSIONS: Regional volume alterations are associated with specific cognitive deficits in schizophrenia. Some structure-cognition relationships differentiate this population from healthy control subjects. PMID- 16039620 TI - The Maudsley Bipolar Disorder Project: executive dysfunction in bipolar disorder I and its clinical correlates. AB - BACKGROUND: Cognitive abnormalities are increasingly recognized as a feature of bipolar I disorder (BDI,) but there is limited information regarding the pattern and severity of cognitive impairment during remission and its relationship with clinical variables. METHODS: Forty-four remitted BDI patients recruited from a representative treatment sample and an equal number of matched healthy volunteers underwent comprehensive clinical and cognitive assessments. Cognitive evaluation covered the domains of IQ, memory, and executive function. The profile of cognitive deficits in patients was examined, and the correlation of executive function with clinical features and treatment variables was explored. RESULTS: Remitted BDI patients were impaired in tests of executive function compared with healthy participants. Within the patient group, current antipsychotic treatment predicted worse performance across all executive function tests, whereas duration of illness predicted loss of inhibitory control. Residual mood symptoms, regardless of polarity, had a negative impact primarily on measures of attentional interference. CONCLUSIONS: These results suggest that impaired executive function might be an important feature of BDI. Antipsychotic treatment, duration of illness, and level of symptoms are the most significant contributors to the observed impairment. PMID- 16039621 TI - Brain serotonin1A receptor binding in major depression is related to psychic and somatic anxiety. AB - BACKGROUND: The anxious phenotype of the 5-HT1A receptor knockout mouse and the anxiolytic properties of 5-HT1A agonists suggest that the 5-HT1A receptor modulates anxiety. We investigated the relationship of anxiety expressed in major depressive disorder (MDD) to regional 5-HT1A binding. METHODS: Positron emission tomography with [carbonyl-11C]WAY-100635 was used to estimate regional 5-HT1A binding potential (BP) in 28 medication-free MDD subjects. Stepwise linear regression assessed the predictive capacity of three anxiety components, derived from a larger MDD sample and termed psychic, somatic, and motoric anxiety, on regional 5-HT1A BP. RESULTS: Higher psychic (beta >or= .63) and lower somatic (beta 80 mM), whereas cereulide in contrast to valinomycin is active already at <1 mM. With cereulide, there is a substantial lag, while valinomycin acts without lag. Both ionophores induce mitochondrial swelling in the presence of K+, in the case of cereulide with a lag. These toxins strongly inhibited respiration at the level of complex IV when used at higher concentrations than that used for detection of ionophoretic transport of K+. At high [KCl] (120 mM), valinomycin was more potent than cereulide both as ionophore and inhibitor, but at low [KCl] (2.5 mM), cereulide was much more potent. Thus, valinomycin needed 20-30 mM KCl for substantial effects, cereulide only 1-3 mM K+, which is close to its level in blood serum. This explains the higher toxicity of cereulide at low concentrations with the positively charged potassium complex being accumulated in the cell by transport through the plasma membrane driven by the membrane potential. Furthermore, with similar concentrations, the final concentration of cereulide in the cells may become higher than that of valinomycin. PMID- 16039681 TI - Sex difference in the principal cytochrome P-450 for tributyltin metabolism in rats. AB - Tributyltin is metabolized by cytochrome P-450 (CYP) system enzymes, and its metabolic fate may contribute to the toxicity of the chemical. In the present study, it is examined whether sex differences in the metabolism of tributyltin exist in rats. In addition, the in vivo and in vitro metabolism of tributyltin was investigated using rat hepatic CYP systems to confirm the principal CYP involved. A significant sex difference in metabolism occurred both in vivo and in vitro, suggesting that one of the CYPs responsible for tributyltin metabolism in rats is male specific or predominant at least. Eight cDNA-expressed rat CYPs, including typical phenobarbital (PB)-inducible forms and members of the CYP2C subfamily, were tested to determine their capability for tributyltin metabolism. Among the enzymes studied, a statistically significant dealkylation of tributyltin was mediated by CYP2C6 and 2C11. Furthermore, the sex difference in metabolism disappeared in vitro after anti-rat CYP2C11 antibody pretreatment because CYP2C11 is a major male-specific form in rats. These results indicate that CYP2C6 is the principal CYP for tributyltin metabolism in female rats, whereas CYP2C11 as well as 2C6 is involved in tributyltin metabolism in male rats, and it is suggested that CYP2C11 is responsible for the significant sex difference in the metabolism of tributyltin observed in rats. PMID- 16039682 TI - O(6)-methylguanine DNA-methyltransferase (MGMT) overexpression in melanoma cells induces resistance to nitrosoureas and temozolomide but sensitizes to mitomycin C. AB - Alkylating agents play an important role in the chemotherapy of malignant melanomas. The activity of alkylating agents depends on their capacity to form alkyl adducts with DNA, in some cases causing cross-linking of DNA strands. However, the use of these agents is limited by cellular resistance induced by the DNA repair enzyme O(6)-methylguanine DNA-methyltransferase (MGMT) which removes alkyl groups from alkylated DNA strands. To determine to what extent the expression of MGMT in melanoma cells induces resistance to alkylating agents, the human cell line CAL77 Mer- (i.e., MGMT deficient) were transfected with pcMGMT vector containing human MGMT cDNA. Several clones expressing MGMT at a high level were selected to determine their sensitivity to chemotherapeutic drugs. Melanoma transfected cells were found to be significantly less sensitive to nitrosoureas (carmustine, fotemustine, streptozotocin) and temozolomide with an increase of IC(50) values between 3 and 14 when compared to parent cells. No difference in cell survival rates between MGMT-proficient and -deficient cells was observed for melphalan, chlorambucil, busulphan, thiotepa and cisplatin which preferentially induce N(7) guanine lesions. Surprisingly, MGMT overexpression increased the sensitivity of CAL77 cells to mitomycin C by approximately 10-fold. Treatment of clonal cell lines with buthionine-[S,R]-sulfoximine (BSO), an inhibitor of gamma glutamylcysteine synthetase which depletes cellular glutathione, completely reversed this unexpected increase in sensitivity to mitomycin C. This observation suggests that glutathione is involved in the sensitivity of MGMT-transfected cells to mitomycin C and may act synergistically with MGMT via an unknown mechanism. PMID- 16039683 TI - Animal carcinogenicity studies on radiofrequency fields related to mobile phones and base stations. AB - Since a report in 1997 on an increased lymphoma incidence in mice chronically exposed to a mobile phone radiofrequency signal, none of the subsequent long-term studies in rodents have confirmed these results. On the other hand, several of the follow-up co- and carcinogenicity studies are still underway or are presently being initiated. Most of the published long-term studies used 1 exposure level only and suffer from a poor dosimetry which does not consider the animal's growth. Additional points of criticism are a limited, in some cases, questionable histopathology and inadequate group sizes. Overall, if dealing with new chemicals or drugs, these studies would not be acceptable for registration with the responsible authorities. The major critical points are taken into consideration within the European co- and carcinogenicity projects (CEMFEC and PERFORM-A), which are in their final stages and in the US long-term studies in mice and rats which are about to be initiated. Nevertheless, the WHO evaluation for health risk assessment of long-term telephone use and base station exposure will start in late 2005. PMID- 16039684 TI - The cytokine-dependent MUTZ-3 cell line as an in vitro model for the screening of contact sensitizers. AB - Langerhans cells (LC) are key mediators of contact allergenicity in the skin. However, no in vitro methods exist which are based on the activation process of LC to predict the sensitization potential of chemicals. In this study, we have evaluated the performances of MUTZ-3, a cytokine-dependent human monocytic cell line, in its response to sensitizers. First, we compared undifferentiated MUTZ-3 cells with several standard human cells such as THP-1, KG-1, HL-60, K-562, and U 937 in their response to the strong sensitizer DNCB and the irritant SDS by monitoring the expression levels of HLA-DR, CD54, and CD86 by flow cytometry. Only MUTZ-3 and THP-1 cells show a strong and specific response to sensitizer, while other cell lines showed very variable responses. Then, we tested MUTZ-3 cells against a wider panel of sensitizers and irritants on a broader spectrum of cell surface markers (HLA-DR, CD40, CD54, CD80, CD86, B7-H1, B7-H2, B7-DC). Of these markers, CD86 proved to be the most reliable since it detected all sensitizers, including benzocaine, a classical false negative in local lymph node assay (LLNA) but not irritants. We confirmed the MUTZ-3 response to DNCB by real time PCR analysis. Taken together, our data suggest that undifferentiated MUTZ-3 cells may represent a valuable in vitro model for the screening of potential sensitizers. PMID- 16039685 TI - Male-mediated developmental toxicity. AB - In recent years, the public has become more aware that exposure of males to certain agents can adversely affect their offspring and cause infertility and cancer. The hazards associated with exposure to ionising radiation have been recognised for nearly a century, but interest was aroused when a cluster of leukaemia cases was identified in young children living in Seascale, close to the nuclear processing plant at Sellafield in West Cumbria. There was a civil court case on behalf of two of the alleged victims of paternal irradiation at Seascale against British Nuclear Fuels. The case foundered on "the balance of probabilities". Nevertheless, there was support for paternal exposure from Japanese experimental X-ray studies in mice. The tumours were clearly heritable as shown by F2 transmission. Also, effects of a relatively non-toxic dose of radiation (1Gy) on cell proliferation transmitted to the embryo were manifested in the germ line of adult male mice even after two generations. In addition in humans, smoking fathers appear to give rise to tumours in the F(1) generation. Using rodent models, developmental abnormalities/congenital malformations and tumours can be studied after exposure of males in an extended dominant lethal assay and congenital malformations can be determined which have similar manifestations in humans. The foetuses can also be investigated for skeletal malformations and litters can be allowed to develop to adulthood when tumours, if present, can be observed. Karyotype analysis can be performed on foetuses and adult offspring to determine if induced genetic damage can be transmitted. Using this study design, cyclophosphamide, 1,3-butadiene and urethane have been examined and each compound produced positive responses: cyclophosphamide in all endpoints examined, 1,3-butadiene in some and urethane only produced liver tumours in F(1) male offspring. This suggests the endpoints are determined by independent genetic events. The results from heritable studies with 1,3-butadiene have been used in the parallelogram approach to determine a risk assessment for the germ cells in man. PMID- 16039686 TI - Worldwide genomic diversity of the human papillomaviruses-53, 56, and 66, a group of high-risk HPVs unrelated to HPV-16 and HPV-18. AB - Among more than 200 human papillomavirus (HPV) types presumed to exist, 18 "high risk" HPV types are frequently found in anogenital cancer. The best studied types are HPV-16 and 18, which are only distantly related to one another and form two separate phylogenetic branches, each including six closely related types. HPV-30, 53, 56, and 66 form a third phylogenetic branch unrelated to HPV-16 and 18. Worldwide comparison of HPV-16 and 18 isolates revealed a distribution of variant genomes that correlated with the geographic origin and the ethnicity of the infected cohort and led to the concept of unique African, European, Asian, and Native American HPV-16 and 18 variants. Here, we address the question whether similar phylogenies are found for HPV-53, 56, and 66 by determining the sequence of the long control regions (LCR) of these HPVs in samples from Europe, Asia, and Africa, and from immigrant societies in North and South America. Phylogenetic trees calculated from point mutations and a few insertions/deletions affecting 2 4.2% of the nucleotide sequences were distinct for each of the three HPVs and divergent from HPV-16 and 18. In contrast to the "star-phylogenies" formed by HPV 16 and 18 variants, 44 HPV-53 isolates represented nine variants, which formed two deep dichotomic branches reminiscent of the beginning split into two new taxa, as recently observed for subtypes of HPV-44 and 68. A total of 66 HPV-56 isolates represented 17 variants, which formed three branches preferentially containing European, Asian, and African variants. Variants of a fourth branch, deeply separated from the other three, were characterized by a 25 bp insertion and created a dichotomy rather than star-like phylogeny. As it contained isolates from cohorts in all continents, it may have evolved before the spread of humans into all continents. 18 of 31 HPV-66 isolates represented the prototype clone, which was found in all parts of the world, while the remaining 13 clones formed 11 branches without any geographic association. Our findings confirm the notion of a quantitatively limited genomic diversity of each HPV type with some correlation to the geographic origin of the sample. In addition, we observed in some variants of these three HPV types mutations that affect the amino acid sequence of the E6 oncoproteins and the L1 capsid protein, supporting the possibility of immunogenic and oncogenic diversity between variants of any HPV type. PMID- 16039687 TI - Antibody response is required for protection from Theiler's virus-induced encephalitis in C57BL/6 mice in the absence of CD8+ T cells. AB - Intracerebral infection of susceptible mice with Theiler's murine encephalomyelitis virus (TMEV) induces immune-mediated demyelinating disease and this system serves as a relevant infectious model for human multiple sclerosis. It was previously shown that beta2M-deficient C57BL/6 mice lacking functional CD8+ T cells display increased viral persistence and enhanced susceptibility to TMEV-induced demyelination, and yet the majority of mice are free of clinical signs. To understand the mechanisms involved in this general resistance of C57BL/6 mice in the absence of CTL responses, mice (muMT) deficient in the B-cell compartment lacking membrane IgM molecules were treated with anti-CD8 antibody and then infected with TMEV. Although little difference in the proliferative responses of peripheral T cells to UV-inactivated TMEV and the resistance to demyelinating disease was observed between virus-infected muMT and control B6 mice, the levels of CD4(+) T cells were higher in the CNS of muMT mice. However, after treatment with anti-CD8 antibody, 100% of the mice displayed clinical gray matter disease and prolonged viral persistence in muMT mice, while only 10% of B6 mice showed clinical symptoms and very low viral persistence. Transfusion of sera from TMEV-infected B6 mice into anti-CD8 antibody-treated muMT mice partially restored resistance to virus-induced encephalitis. These results indicate that the early anti-viral antibody response is also important in the protection from TMEV-induced encephalitis particularly in the absence of CD8+ T cells. PMID- 16039688 TI - Host range mutants of Minute Virus of Mice with a single VP2 amino acid change require additional silent mutations that regulate NS2 accumulation. AB - Two host range switch mutants of the immunosuppressive strain of parvovirus Minute Virus of Mice (MVMi) were isolated from plaques on A9 fibroblasts. Both carried a single coding mutation at residue D399 in VP2, to alanine and glycine in hr105 and hr107, respectively, and a second, non-coding, guanine-to-adenine change at nucleotide 1970 in hr105 and 1967 in hr107. These mutations were recreated in a wild type MVMi infectious plasmid clone, both alone and as pairs, in either the original or switched combinations. All single mutants failed to replicate productively in fibroblasts, but the two pairs of changes were functionally equivalent. Single D399 mutations allowed the viruses to initiate infection in fibroblasts, but NS2 expression was severely restricted and correlated with poor accumulation and release of progeny virus. Mutations at 1967 or 1970 enhanced NS2 accumulation, and allowed efficient progeny production and release. Conversely, the D399 mutations destroyed the viruses' ability to infect EL4 lymphocytes. In all productive EL4 infections, NS2 was expressed at high ratios even in the absence of upstream mutations, and progeny accumulation was efficient. However, EL4 cells lack a mechanism for early progeny release, potentially explaining why virus amplification in these cells is slow. PMID- 16039689 TI - Why is second-order vision less efficient than first-order vision? AB - Research has shown that the sensitivity to second-order modulations of carrier contrast is lower than that to first-order luminance modulations stimuli. We sought to compare the efficiency of processing first- and second-order information. Employing a phase-discrimination paradigm we found that when humans were given sufficient a priori information of signal parameters they detected both luminance and contrast modulations of 0.6 and 2c/deg by a phase-sensitive algorithm. The overall detection efficiency for second-order patterns, however, was lower that that for first-order stimuli. To study the factors which limit the efficiency of first- and second-order vision, we measured detection performance for luminance and contrast modulations of 0.6 and 2c/deg embedded in Gaussian noise. The results showed that the detection of second-order patterns had lower sampling efficiency and higher additive internal noise as compared to the detection of first-order stimuli. Classification images for detecting contrast modulations of 2c/deg resembled the side-band component of the contrast modulations which suggests that human observers may detect contrast modulations of a sinusoidal carrier using first-order luminance channels. The lower sensitivity of the mechanism detecting second-order patterns might be due to higher levels of additive internal noise and lower sampling efficiency than those of the mechanism analysing first-order patterns. PMID- 16039690 TI - Attentive tracking shifts the perceived location of a nearby flash. AB - Several studies have shown that the perceived position of a briefly presented stimulus can be displaced by nearby motion or by eye movements. We examined whether attentive tracking can also modulate the perceived position of flashed static objects when eye movements and low-level motion are controlled. Observers attentively tracked two target bars 180 degrees apart on a rotating, 12-spoke radial grating and judged the alignment of two flashes that were briefly presented, one on each side of the grating. Because of the symmetry of the 12 spoke grating, test flashes could be timed so that the rotating grating was always aligned to a standard orientation at the time of the test, while the tracked bars themselves, being only two of the 12 spokes, could probe locations that differed by multiples of 30 degrees ahead of, aligned with, or behind, the test bars. Despite the physical identity of the stimulus in each test--same orientation, same motion--the perceived position of the two flashes strongly depended on the locus of attention: when the test flashes were presented ahead of the tracked bars, a large position shift in the direction of the grating's motion was seen. If they were presented behind the tracked bars, the illusory displacement was reduced or slightly reversed. These effects of attention led us to suggest an attentional model of position distortions that links the effects seen for motion and for eye movements. PMID- 16039691 TI - Contribution of retinal neurons to d-wave of primate photopic electroretinograms. AB - The purpose of this study was to determine the contribution of different types of retinal neurons to the d-wave of the primate electroretinogram using pharmacological agents. NMDA + TTX was used to suppress inner retinal activity, and APB and PDA to block the activity of the ON- and OFF-pathways, respectively. Results indicated that the inner retinal neurons had a small but certain contribution to the d-wave. The initial rapid phase of the d-wave originates from the activity of the cone OFF-pathway nearly exclusively, and the later slow phase is shaped by the cone photoreceptors. The cone ON-pathway acts in a direction opposite to that of the other components. PMID- 16039692 TI - Repetitive TMS over posterior STS disrupts perception of biological motion. AB - Biological motion perception, the recognition of human action depicted in sparse dot displays, is supported by a network of brain areas including the human posterior superior temporal sulcus (pSTS). We have used repetitive transcranial magnetic stimulation (rTMS) to temporarily disrupt cortical activity within the pSTS and subsequently measured sensitivity to biological motion. Sensitivity was measured for canonical (upright) point-light animations and for animations inverted 180 deg, a manipulation that renders biological motion more difficult to recognize. Observers were markedly less sensitive to upright biological motion following pSTS stimulation. In contrast, performance remained normal for inverted biological motion following pSTS stimulation, and normal for upright and inverted biological motion following stimulation over visual motion sensitive area MT+/V5. In connection with previous brain imaging results, our findings demonstrate that normal functioning of the posterior STS is required for intact perception of biological motion. PMID- 16039693 TI - Eutrophication downstream from small reservoirs in mountain rivers of Central Spain. AB - In this research we examined the hypothesis that upper reaches of rivers and streams can experience eutrophication as a consequence of deep releases from dams. Field studies were conducted in four mountain rivers (Tormes, Riaza, Eresma and Miraflores Rivers) of Central Spain. The watersheds of these rivers are underlain by siliceous rocks. A small deep-release storage reservoir is found in the upper reaches of each river. Two sampling sites, upstream and downstream from the reservoir, were established in stony riffles of each impounded river. Significant (P < 0.01) increases in conductivity and nutrient (NO3-N, NH4-N, PO4 P) concentrations downstream from the reservoirs were measured. Significant (P < 0.01) increases in periphyton chlorophyll a and ash-free dry biomass were also quantified at downstream sites. Significant (P < 0.01) correlation coefficients indicated that phosphate would play a more important role as the limiting nutrient for periphyton. Relative abundances of macroinvertebrate scrapers and collector-gatherers increased downstream from the reservoirs. Furthermore, taxon dominance, total density and total biomass of macroinvertebrates tended to be higher at downstream sites than at upstream sites. In contrast, taxon diversity and relative abundance of macroinvertebrate shredders decreased downstream from the reservoirs. It is concluded that small deep-release storage reservoirs, located in upper reaches of siliceous rivers, can act as nutrient sources, causing eutrophication downstream. Nutrients would ultimately come from land/forest runoff. The fact that terrestrial vegetation was not completely removed before filling reservoirs could also contribute to the eutrophication process. PMID- 16039694 TI - Biodegradability of the X-ray contrast compound diatrizoic acid, identification of aerobic degradation products and effects against sewage sludge micro organisms. AB - Pharmaceuticals and contrast media have been detected in hospital effluents, sewage treatment plants, surface water, and ground water. Only little is known about their elimination during sewage treatment and effects of possible biotransformation products against bacteria. The modified Zahn-Wellens test (ZWT, OECD 302 B) and a test simulating biological sewage treatment (modified OECD 303 A test) were used to assess the biodegradability of the widely used ionic iodinated contrast agent diatrizoic acid (diatrizoate). Effects against sewage sludge bacteria were studied in the two test systems by monitoring the biomarkers quinones, polyamines, phospholipids and adenosine triphosphate. Diatrizoate was biotransformed into 2,4,6-triiodo-3,5-diamino-benzoic acid in the ZWT. 2,4,6 Triiodo-3,5-diamino-benzoic acid was stable under the test conditions of the ZWT. Diatrizoate was not eliminated in the OECD 303 A simulation test. It was not adsorbed by the sewage sludge. No effects of the test compound or its aerobic transformation products against the bacteria present in the sewage sludge were detected using phospholipids, quinones, polyamines, and adenosine triphosphate as biomarkers. PMID- 16039695 TI - Immobilization of heavy metals in polluted soils by the addition of zeolitic material synthesized from coal fly ash. AB - The use of zeolitic material synthesized from coal fly ash for the immobilization of pollutants in contaminated soils was investigated in experimental plots in the Guadiamar Valley (SW Spain). This area was affected by a pyrite slurry spill in April 1998. Although reclamation activities were completed in a few months, residual pyrite slurry mixed with soil accounted for relatively high leachable levels of trace elements such as Zn, Pb, As, Cu, Sb, Co, Tl and Cd. Phytoremediation strategies were adopted for the final recovery of the polluted soils. The immobilization of metals had previously been undertaken to avoid leaching processes and the consequent groundwater pollution. To this end, 1100 kg of high NaP1 (Na6[(AlO2)6(SiO2)10] .15H2O) zeolitic material was synthesized using fly ash from the Teruel power plant (NE Spain), in a 10 m3 reactor. This zeolitic material was manually applied using different doses (10000-25000 kg per hectare), into the 25 cm topsoil. Another plot (control) was maintained without zeolite. Sampling was carried out 1 and 2 years after the zeolite addition. The results show that the zeolitic material considerably decreases the leaching of Cd, Co, Cu, Ni, and Zn. The sorption of metals in soil clay minerals (illite) proved to be the main cause contributing to the immobilization of these pollutants. This sorption could be a consequence of the rise in pH from 3.3 to 7.6 owing to the alkalinity of the zeolitic material added (caused by traces of free lime in the fly ash, or residual NaOH from synthesis). PMID- 16039696 TI - Evaluation of clopidogrel resistance. PMID- 16039697 TI - Prothrombotic abnormalities in childhood ischaemic stroke. AB - Childhood ischaemic stroke, incorporating arterial ischaemic stroke and cerebral sinus venous thrombosis, is associated with significant morbidity and mortality in children. The majority of cases in children present with well-recognised risk factors. The appreciation of the role prothrombotic abnormalities have in disease states is developing rapidly. Prothrombotic abnormalities are abnormalities of the coagulation system, fibrinolytic system, endothelial cells or platelets that lead to a reduced threshold for pathological thrombus formation. Our understanding of the role of prothrombotic abnormalities in childhood ischaemic stroke is increasing and has a direct bearing on the development of effective management and prevention strategies. We provide a brief background of prothrombotic abnormalities and review the available literature on prothrombotic markers in childhood ischaemic stroke. Overall, prothrombotic abnormalities have been identified in 20-50% of children presenting with AIS and 33-99% of children with cerebral sinus venous thrombosis. There appear to be a number of associations emerging including an increased frequency of factor V Leiden mutation, elevated lipoprotein (a), protein C deficiency and antiphospholipid antibodies in children presenting with arterial ischaemic stroke. The pathogenic role of prothrombotic abnormalities as predisposing to initial and recurrent childhood ischaemic stroke is becoming increasingly evident. The impact on treatment, however, will only be clarified with carefully designed, multi institutional prospective studies. PMID- 16039698 TI - Thromboembolic complications in children with cancer. AB - Thrombosis is a significant problem in patients with cancer. The impact of thromboembolism in association with childhood cancer is not clearly defined. Similarly the information of prevention and management of thromboembolic events in children receiving cancer-therapy is limited. This review aims to examine current knowledge regarding the epidemiology, pathophysiology and management of thrombosis in association with cancer in children. PMID- 16039699 TI - Implications of second-look laparotomy in the context of optimally resected stage III ovarian cancer: a non-randomized comparison using an explanatory analysis: a Gynecologic Oncology Group study. AB - OBJECTIVE: A non-randomized comparison of outcome in women undergoing second-look laparotomy (SLL) or clinical follow-up, after receiving six cycles of combination chemotherapy with paclitaxel plus either cisplatin or carboplatin, for optimally resected stage III ovarian cancer. METHODS: Prior to chemotherapy randomization, patients chose whether or not to undergo SLL; this was a stratification factor to insure balance of treatment assignment. Any subsequent therapy was physician directed. Explanatory analysis replaced intent-to-treat because of a higher likelihood of detecting SLL effect in the presence of noncompliance. RESULTS: There were 393 patients (median age: 54) who Elected SLL and 399 (median age: 59) who Elected No SLL. The former group was more likely to have gross residual disease at initial surgery than the latter group (69% versus 60%, respectively). In the Elected SLL group, 59 (15%) patients subsequently refused surgery, in nine (2%) surgery was contraindicated, and 31 (8%) relapsed or died prior to the procedure. Cancer was found in 46% of 294 (75%) patients undergoing SLL. Since early failures (prior to SLL) do not address benefit, such patients (SLL: 32; No SLL: 33), defined as progression-free survival (PFS) < 6 months, were excluded from analysis. The adjusted relative risk of progression is 0.89 (95% confidence interval: 0.75, 1.07); the difference in median PFS is 1.0 month (SLL: 23.9 months; No SLL: 22.9 months). The survival rate curves are superimposable. CONCLUSION: In the context of a non-randomized comparison, the performance of a SLL was not associated with longer survival. PMID- 16039700 TI - Differential effects of mild repeated restraint stress on behaviors and GABA(A) receptors in male and female rats. AB - We previously reported that the very mild stress of individual housing influenced seizure risk and gamma-amino butyric acid (GABA(A)) receptor activity differentially between male and female rats. The aim of the present set of studies was to assess sex differences in behavioral responses to a more pronounced type of stressor, repeated restraint stress. We also wanted to determine the role of GABA(A) receptors in effects of this stressor. Our data suggest that repeated restraint stress afforded short-term protection against seizure induction in both male and female rats. Moreover, this protection was more persistent in female than male rats. This stress paradigm also elicited a reduction in general activity in male rats, whereas female rats displayed prolonged increased activity following the repeated restraint stress exposure. However, there were limited effects on anxiety-like behaviors, as determined by time spent in the open arms on the elevated plus maze. Sex differences in stress induced increases in plasma corticosterone levels were observed, which generally correlated with sex differences in behavioral measures. There were no significant effects of the repeated restraint stress exposure on benzodiazepine/GABA(A) receptor density or affinity nor on receptor function. Taken together, these findings provide additional evidence to support the important influences of sex in responding to stress and highlight the need to consider this context when addressing the role of stress in health issues for women and men. PMID- 16039701 TI - Seroconversion of calves following intravenous injection with embryos exposed to bovine viral diarrhea virus (BVDV) in vitro. AB - Two recent studies demonstrated that a high-affinity isolate of BVDV (SD-1), remained associated with a small percentage of in vivo-derived bovine embryos following artificial exposure to the virus and either washing or trypsin treatment. Further, the embryo-associated virus was infective in an in vitro environment. Therefore, the objective of this study was to determine if the quantity of a high-affinity isolate of BVDV associated with single-washed or trypsin-treated embryos could cause infection in vivo. Twenty zona-pellucida intact morulae and blastocysts (MB) were collected on day 7 from superovulated cows. After collection, all MB were washed according to International Embryo Transfer Society (IETS) standards, and all but 4 MB (negative controls) were exposed for 2 h to 10(5)-10(6) cell culture infective doses (50% endpoint) per milliliter (CCID(50)/mL) of viral strain SD-1. Following exposure, according to IETS standards, one half of the MB were washed and one half were trypsin treated. All MB were then individually sonicated, and sonicate fluids were injected intravenously into calves on day 0. Blood was drawn to monitor for viremia and(or) seroconversion. Seroconversion of calves injected with sonicate fluids from washed and trypsin-treated embryos occurred 38% and 13% of the time, respectively. Therefore, the quantity of a high-affinity isolate of BVDV associated with single-washed or trypsin-treated embryos was infective in vivo. PMID- 16039702 TI - Resynchronization of previously timed-inseminated beef heifers with progestins. AB - The objective was to determine the efficacy of a previously used CIDR or melengestrol acetate (MGA; 0.5mg/head/day) for resynchronization of estrus in beef heifers not pregnant to timed-AI (TAI). In three experiments and a field trial, heifers were reinseminated 6-12 h after first detection of estrus. Pregnancy diagnosis was done from approximately 25-43 days after either TAI or reinsemination. In Experiment 1, 79 heifers received a once-used CIDR from 13 to 20 days after TAI and 80 heifers were untreated controls. For these two groups, there were 34 and 35 heifers, respectively, not pregnant to TAI; median +/- S.E. intervals from TAI to onset of estrus were 22 +/- 0.2 days versus 20 +/- 0.6 days (P < 0.001); estrus rates were 70.6% versus 85.7% (P = 0.1); conception rates were 62.5% versus 76.7% (P < 0.3); and pregnancy rates were 44.1% versus 65.7% (P = 0.07), for CIDR and untreated (control) groups, respectively. In Experiment 2, heifers (n = 651) were TAI (Day 0) and 13 days later randomly assigned to one of seven groups (n = 93 per group) to receive a once-used CIDR (three groups; Days 13-20), MGA (three groups; Days 13-19), or no treatment (control group). Groups given a CIDR or MGA also received: no further treatment (CIDR or MGA alone); 1.5mg estradiol-17beta (E-17beta) and 50 mg progesterone (P4) in 2 mL canola oil on Day 13; or E-17beta and P4 on Day 13 and 0.5 mg E-17beta on Day 21 (24 h after CIDR removal or 48 h after the last feeding of MGA). Pregnancy rate to TAI was lowest (P < 0.05) for the group given a CIDR plus E-17beta and P4 on Day 13 and E 17beta on Day 21. Variability in return to estrus was greater (P < 0.001) in the control and MGA groups than in CIDR groups. Conception and pregnancy rates in heifers given a CIDR (65.1 and 61.4%) were higher (P<0.01) than those fed MGA (49.6 and 40.4%), but not different from controls (62.2 and 54.9%, respectively). In Experiment 3, 616 heifers received a once- or twice-used CIDR for 7 days, beginning 13+/-1 days after TAI, with or without a concurrent injection of 150 mg of P4 (2 x 2 factorial design). Pregnancy rate to TAI was 47.2%. In heifers that returned to estrus, there was no significant difference between a once- or twice used CIDR for rates of estrus (68.8%, P < 0.3), conception (65.9%, P < 0.6) and pregnancy (45.3%, P < 0.8). Injecting progesterone at CIDR insertion increased the median interval from CIDR removal to onset of estrus (P < 0.05) and reduced rates of estrus (63.8% versus 73.8%, P<0.05), conception (60.5% versus 70.6%, P = 0.1) and pregnancy (38.6% versus 52.2%, P < 0.02). In a field trial, 983 heifers received a once-used CIDR for 7 days, beginning 13 +/- 1 days after TAI. Pregnancy rate to TAI was 55.2%. The median (and mode) of the interval from CIDR removal to estrus was 2.5 days. Estrus, conception and pregnancy rates were 78.2, 70.3 and 55.0% (overall pregnancy rate to TAI and rebreeding, 78.7%). In summary, a once- or twice-used CIDR for 7 days, starting 13 +/- 1 days after TAI resulted in the majority of nonpregnant heifers detected in estrus over a 4-day interval, with acceptable conception rates; however, injecting progesterone at CIDR insertion significantly reduced both estrus and pregnancy rates, and estradiol treatment after CIDR removal was associated with a decreased pregnancy rate to TAI. Fertility was higher in heifers resynchronized with a once-used CIDR than with MGA. PMID- 16039703 TI - Film thickness and fluidity of various luting cements determined using a trial indentation meter. AB - OBJECTIVES: The purpose of this study was to evaluate a new trial apparatus which could measure film thickness (T(h)) under a low load, using an indenter. METHODS: Five commercial luting cements were measured using this apparatus. T(h) was determined using four kinds of indenter, which were 5.0, 10.0, 15.0 and 20.0mm in diameter. For each indenter, the load per contact area of the indenter with the platen equaled 150N per 200 mm2 described in ISO No. 9917. The effects of working time (30, 60, and 90 s) and load (2.86, 6.78, 10.7, and 14.7N) on the fluidity and T(h) were examined using the indenter with a diameter of 5.0mm. The duration of loading was 10 min. The fluidity was defined as the reciprocal of the apparent viscosity and was calculated using Stefan's equation. RESULTS: The T(h) obtained using this apparatus agreed with the film thickness obtained using the test described in ISO No. 9917. For Durelon (DU), T(h) increased 1.58 times and fluidity decreased 0.26 times as the working time increased from 30 to 90 s; T(h) decreased 0.56 times and fluidity increased 1.28 times as loading increased from 2.86 to 14.7N. For Ketac cement radiopaque (KE), Panavia fluoro cement (PA), Scotchbond resin cement (SC), and Vitremer luting cement (VI), although the fluidity and T(h) were not influenced much by duration (30-90 s), T(h) decreased 0.79, 0.74, 0.95, and 0.78 times for KE, PA, SC, and VI, respectively, and fluidity decreased 0.68, 0.51, 0.30, and 0.36 times, respectively, as loading increased from 2.86 to 14.7N. SIGNIFICANCE: This apparatus was proven effective in elucidating the flow properties of luting cement. PMID- 16039704 TI - Dynamic mechanical thermal analysis of two light-cured dental composites. AB - OBJECTIVES: Clinical observations suggest that some composite resins are more often linked to post-operative sensitivity than others. These differences may relate to differences in modulus of elasticity and polymerization rates among materials. The aim of this study was to identify viscoelastic behavior of two light curable composites and determine whether significant differences in viscoelastic behavior exist between the two materials when light cured at each of three different irradiance values. METHODS: Two composites (Z100 and Z250 by 3M ESPE) were evaluated. Six specimens per composite and irradiance value (250, 500 and 850 mW/cm2) were made. The curing times were chosen to produce a fixed energy value of 30 J/cm2 independent of irradiation value. Dynamic mechanical thermal analysis (DMTA) was performed in single cantilever clamped mode. RESULTS: There were significant differences in transition temperatures between the two materials and the three frequencies at their glass transition temperatures, while significant differences did not exist at the lower transitions. The glass transition of Z250 was lower and narrower than that of Z100. Z250 exhibited lower storage modulus values. The irradiance values did not affect any of the transition temperatures significantly. SIGNIFICANCE: The lower and more distinct T(g) of Z250 suggests that Z250 cures more efficiently than Z100. The lower storage modulus of Z250 suggests that Z250 develops less stress in the tooth than Z100 during curing if shrinkage is the same for the two materials. The findings suggest that the material chosen, rather than irradiance, determines the stress level developed during light curing. PMID- 16039705 TI - Microtensile bond strength of a resin cement to glass infiltrated zirconia reinforced ceramic: the effect of surface conditioning. AB - OBJECTIVES: This study evaluated the effect of three surface conditioning methods on the microtensile bond strength of resin cement to a glass-infiltrated zirconia reinforced alumina-based core ceramic. METHODS: Thirty blocks (5 x 5 x 4 mm) of In-Ceram Zirconia ceramics (In-Ceram Zirconia-INC-ZR, VITA) were fabricated according to the manufacturer's instructions and duplicated in resin composite. The specimens were polished and assigned to one of the following three treatment conditions (n=10): (1) Airborne particle abrasion with 110 microm Al(2)O(3) particles + silanization, (2) Silica coating with 110 microm SiO(x) particles (Rocatec Pre and Plus, 3M ESPE) + silanization, (3) Silica coating with 30 microm SiO(x) particles (CoJet, 3M ESPE) + silanization. The ceramic-composite blocks were cemented with the resin cement (Panavia F) and stored at 37 degrees C in distilled water for 7 days prior to bond tests. The blocks were cut under coolant water to produce bar specimens with a bonding area of approximately 0.6mm(2). The bond strength tests were performed in a universal testing machine (cross-head speed: 1mm/min). The mean bond strengths of the specimens of each block were statistically analyzed using ANOVA and Tukey's test (alpha0.05) were found between the tensile strength of the posts luted with zinc phosphate (414+/-102N) and the resin composite cements. Silica coating and silanization revealed the highest tensile bond strength in posts luted with Compolute Aplicap but it was not effective for the other experimental groups. Zinc phosphate cement exhibited tensile bond strength as good as resin composite cements. PMID- 16039708 TI - The automatic detection of known beta-propeller structural motifs from protein tertiary structure. AB - Following our previous work on the analysis of 'structural plasticity' associated with the beta-propeller structural motifs, we have now developed a simple method that can automatically detect all the known beta-propellers in protein tertiary structure, given a list of Protein Data Bank (PDB) codes as input to the computer program. Our beta-propeller detection (BPD) method identifies the location of beta-propellers in the protein structure, specifies the beta-propeller type, the beta-sheet associated beta-strand pattern and the structurally similar beta propellers observed in other proteins. When tested on 21,566 proteins in the PDB, the BPD method was capable of correctly identifying all the known 245 beta propellers described in the structural classification of proteins (SCOP) with the number of false positives detected being less than 0.2%. Forty-one false positives were detected that correspond to eight known protein families. When compared with some of the popular web-based programs that can automatically detect 'structural similarities' between the query and target proteins, our method has the advantage of also being capable of detecting beta-propellers associated with 'structural plasticity' and in situations where the target and query proteins differ in amino acid sequence length. PMID- 16039709 TI - A PLGA membrane controlling cell behaviour for promoting tissue regeneration. AB - Barrier membranes are used in periodontal applications with the aim of supporting bone regeneration by physically blocking migrating epithelial cells. We report a membrane design that has a surface topography that can inhibit epithelial cell migration and proliferation on one side and a topography that guides osteoblast migration to a desired area. A PLGA copolymer (85:15) blended with MePEG, was cast to have surfaces with smooth, grooved or sandblasted-acid-etched topographies. Epithelial cells spread on smooth surfaces after 24 h, and cell numbers increased after 5 days. Cells on the smooth surface exhibited no preferred direction of migration. On the sandblasted-acid-etched topography epithelial cells spread but the cell number did not significantly increase after 5 days. Cell migration was inhibited on this surface. Osteoblasts spread on both grooved and smooth surfaces and cell number increased after 5 days on all surfaces. The cells that adhered in the grooves migrated preferentially in the direction of the grooves. Positive alkaline phosphatase staining was seen on all surfaces within 4 weeks and positive Von Kossa nodule staining within 6 weeks. These results suggest that surface topographies replicated on opposite sides of a biodegradable polymers membrane can inhibit proliferation and migration of the epithelial cells, and promote proliferation and directional migration of osteoblasts. Addition of appropriate surface topographies to membranes used in guided tissue regeneration has the possibility of improving clinical performance in periodontal tissue regeneration procedures. PMID- 16039710 TI - Tissue engineering of human cartilage and osteochondral composites using recirculation bioreactors. AB - Chondrocytes isolated from human foetal epiphyseal cartilage were seeded dynamically into polyglycolic acid (PGA) scaffolds and cultured in recirculation column bioreactors to produce tissue-engineered cartilage. Several culture techniques with the potential to provide endogenous growth factors and other conditions beneficial for de novo cartilage synthesis were investigated. Osteochondral composite constructs were generated by seeding separate PGA scaffolds with either foetal chondrocytes or foetal osteoblasts then suturing the scaffolds together before bioreactor cultivation. This type of co-culture system provided direct contact between the tissue-engineered cartilage and developing tissue-engineered bone and yielded significant improvements in cartilage quality. In the cartilage section of the composites, the concentrations of glycosaminoglycan (GAG) and total collagen were increased by 55% and 2.5-fold, respectively, compared with control cartilage cultures, while levels of collagen type II were similar to those in the controls. The osteochondral composites were harvested from the bioreactors as single units with good integration between the cartilage and bone tissues. Only the cartilage layer contained GAG while only the bone layer was mineralised. In other experiments, co-culture of tissue-engineered cartilage with pieces of ex-vivo cartilage or ex-vivo bone did not improve the quality of the cartilage relative to control cultures. Addition of 10(-6) M diacerein to the culture medium also had no effect on the properties of engineered cartilage. This work demonstrates the beneficial effects of generating cartilage tissues in contact with developing bone. It also demonstrates the feasibility of producing composite osteochondral constructs for clinical application using recirculation column bioreactors. PMID- 16039711 TI - Topical delivery of low-molecular-weight heparin with surface-charged flexible liposomes. AB - To increase topical delivery of low-molecular-weight heparin (LMWH), cationic, neutral, and anionic flexible liposomes (cFlexosome, nFlexosome, and aFlexosome) were prepared. The effects of surface charge of Flexosome on physicochemical properties and skin penetration of LMWH were also investigated. Among the different formulations of Flexosome, cFlexosome demonstrated three-times higher entrapment efficiency of LMWH, and better physicochemical stability than nFlexosome and aFlexosome. In vitro skin penetration and in vivo localization into the deeper skin layer of LMWH were significantly greater from cFlexosome compared to other formulations. Changes of skin surface charge after LMWH cFlexosome application were investigated as a function of time. In the process of skin penetration, the Flexosomes act as drug carrier with the associated LMWH. Overall, macromolecular LMWH could be delivered deeply into the skin by topical application of cFlexosome for the treatment of superficial thrombosis, subcutaneous wounds, bruise, and burns. PMID- 16039712 TI - In vivo behavior of acrylic bone cement in total hip arthroplasty. AB - Polymethylmethacrylate (PMMA) bone cement serves as the primary fixation material between bone and the prosthetic component in cemented total hip arthroplasty. In vivo degradation of bone cement may lead to a decrease in mechanical properties of PMMA and result in aseptic loosening. However, other factors such as porosity and location of the cement relative to the bone implant interface may also contribute to mechanical behavior in vivo. This study investigated the mechanical properties of Simplex cement retrieved from 43 patients undergoing revision total hip arthroplasty. The time in vivo was between 1 month and 27 years. The variables studied included fracture toughness (KIC), porosity, molecular weight, time in vivo of the cement, and relative in vivo location of the cement with respect to the implant and bone. KIC did not correlate with time in vivo of the samples or with molecular weight. This suggests that time in vivo may not be the limiting factor in the mechanical integrity of the bone cement, A significant and inverse relationship was found between porosity and KIC. This implies that porosity is the most important factor in the mechanical behavior of bone cement during in vivo use. PMID- 16039713 TI - Decreased numbers of CD4+ T lymphocytes in peripheral blood after treatment of childhood acute lymphoblastic leukemia. AB - Treatment-related immunosuppression in patients with acute lymphoblastic leukemia (ALL) is associated with increased susceptibility to infectious diseases, also after the treatment. The aim of the present study was the detailed evaluation of T lymphocyte subsets in peripheral blood in children after treatment of ALL. All children were treated according to the BFM 90 protocol. The patients were divided into 5 groups of 30 children in each, depending on the time from cessation of the ALL treatment. A control group consisted of 30 healthy children subjected to elective "1-day" surgery. The children's age ranged from 6 to 18 years. The examinations were performed in FACScan flow cytometer with the use of wide set of monoclonal antibodies: CD3, CD4, CD8, TCRalphabeta, TCRgammadelta, CD19, CD25, CD45RA, CD45RO, CD69, HLA-DR, CD16 and CD56, which particularly allowed detailed analysis of T lymphocytes. The results showed that most parameters in children 1 year after ALL treatment completion were similar to healthy children. However, we observed persistently low CD4+ T cell numbers, both CD45RA+ as well as CD45RO+ subsets as compared to the control group. This might reflect decreased regenerative potential of immunological system in children 1 year after ALL treatment. PMID- 16039714 TI - Chronic lymphocytic leukemia cell CD38 expression and inducible nitric oxide synthase expression are associated with serum IL-4 levels. AB - B cell chronic lymphocytic leukemia (CLL) CD38 expression is variable and may predict outcome. Inducible nitric oxide synthase (NOS2) expression regulates CLL cell apoptosis. IL-4 and IFN-gamma regulate B cell CD38 expression and NOS2 expression. We compared IL-4 and IFN-gamma serum levels between CLL patients and normal individuals, and determined whether serum IL-4 and IFN-gamma levels correlated with CLL cell CD38 expression and NOS enzyme activity. IL-4 levels, but not IFN-gamma levels, differed between normal individuals and CLL patients. Furthermore, there was an association of IL-4 levels, but not IFN-gamma levels, with CD38 and NOS2 expression in CLL patients. PMID- 16039715 TI - Aberrant methylation in promoter-associated CpG islands of multiple genes in acute lymphoblastic leukemia. AB - Methylation profile was analyzed in 10 childhood acute lymphoblastic leukemia (ALL) and nine adult ALL cases. Four genes (p15, p16, RARbeta, FHIT) had methylation in both diseases, four genes (p14, Rb, MLH1, DAPK) showed no methylation in both diseases, and the two genes (APC, RIZ) demonstrated methylation only in adult ALL. Methylation of the RARbeta was more frequent in adult ALL than that in childhood ALL (p=0.01). The number of patients with methylation of multiple genes was higher in adult ALL than that in childhood ALL (p=0.006). Moreover, overall frequency of methylation was higher in adult ALL than that in childhood ALL (p=0.01). PMID- 16039716 TI - Incidence of open-angle glaucoma in a general elderly population: the Rotterdam Study. AB - PURPOSE: To determine the incidence of open-angle glaucoma (OAG) in a general elderly white population. DESIGN: Population-based cohort study. PARTICIPANTS: Participants 55 years and older from the Rotterdam Study, The Netherlands. METHODS: Participants at risk for incident OAG underwent at baseline (1990-1993) and follow-up (1997-1999) the same ophthalmic examination, including measurement of intraocular pressure, visual field testing, and stereo optic disc photography. The diagnosis of probable or definite OAG was made with an algorithm based on optic disc parameters and on visual field testing, independent of the intraocular pressure. MAIN OUTCOME MEASURES: Five-year risk and incidence rates of OAG. RESULTS: In total, 3842 participants were examined (participation rate at follow up, 78%). After a mean follow-up time of 6.5 years, probable (n = 58) or definite OAG (n = 29) developed in 87 persons. The 5-year risk of probable OAG was 1.2% and that of definite OAG 0.6%, and the rate rose for both together from 1% at age 60 years to approximately 3% at age 80 years. Bilateral OAG occurred 5 times more often after than before age 75 years. In participants with prevalent OAG in 1 eye, the 5-year risk of OAG in the fellow eye was 5 times higher than in fellow eyes of non-OAG eyes. Only 37% of the incident cases received treatment for OAG at the time of the reexamination. CONCLUSIONS: The incidence of OAG rose significantly with age. Most of the patients with incident OAG were unaware of having OAG. PMID- 16039717 TI - Glaucoma management among individuals enrolled in a single comprehensive insurance plan. AB - OBJECTIVE: To determine the management patterns for glaucoma and suspect glaucoma in a nationally representative sample of newly treated persons. DESIGN: Retrospective cohort study of persons enrolled in a large managed care organization. PARTICIPANTS: One thousand seven hundred twelve diagnosed suspects and 3623 diagnosed glaucoma patients. METHODS: Linked pharmacy and patient care data were used to examine the glaucoma management and treatment patterns in this cohort of persons insured by a single managed care organization. Rates of monitoring and treatment were calculated for the 3 study groups. MAIN OUTCOME MEASURES: Probability of monitoring (return visits, visual fields [VFs], and optic nerve head imaging or photography) and treatment (argon laser trabeculoplasty [ALT] and surgery) for newly treated persons with suspect and glaucoma diagnoses. RESULTS: After a median follow-up of 440 days, 83% of treated diagnosed suspects had had a billed follow-up office visit to either an optometrist or an ophthalmologist at any time during follow-up, 46% had had at least one billed VF, and 13% had had some form of optic nerve head imaging. Rates were slightly higher for those with diagnosed glaucoma (P>0.05). Surgery and ALT were performed rarely in this treated population (1%-6% at 2 years). CONCLUSIONS: This study suggests that a large proportion of individuals felt to require treatment for glaucoma or suspect glaucoma are falling out of care and are being monitored at rates lower than expected from recommendations of published guidelines. More research is needed to confirm these findings and to determine the reasons for loss to follow-up and low monitoring rates. PMID- 16039718 TI - Optical coherence tomography characteristics of choroidal metastasis. AB - PURPOSE: To report 4 patients (7 eyes) with choroidal metastasis (primary: breast [2], lung [1], and unknown [1]) visualized with optical coherence tomography (OCT). DESIGN: Retrospective case series. PARTICIPANTS: Four patients (7 eyes) with choroidal metastasis participated in the study. METHODS: Clinical examination, ultrasonography, fluorescein angiography, and OCT were performed in all cases. MAIN OUTCOME MEASURES: Optical coherence tomography characteristics. RESULTS: Three patients suffered a gradual onset of blurred vision in both eyes. The fourth presented a unilateral decrease in visual acuity. Bilateral choroidal lesions with various degrees of exudative retinal detachment (RD) were present in 3 of our 4 patients, and there was 1 case of a unilateral unifocal choroidal lesion. Optical coherence tomography revealed anterior displacement of the photoreceptor layer by subretinal fluid (hyporeflective space) overlying a hyperreflective thickened retinal pigment epithelium-choriocapillaris complex. Subretinal deposits with several degrees of hyperreflectivity were seen, as well as loss of normal retinal architecture with intraretinal splitting, identified as regions of low reflectance within the neurosensory retina. After chemotherapy, OCT demonstrated improvement of the serous RD and reduction of the lesion's size with recovery of the normal retinal architecture. CONCLUSIONS: Optical coherence tomography in patients with choroidal metastasis allows evaluation of secondary retinal-retinal pigment epithelial changes. In addition, the technique is useful in the follow-up of lesions after treatment. However, some limitations result from the choroidal location of the metastasis. Further studies with clinicopathological correlation are needed to confirm our observations. PMID- 16039719 TI - Linear scan voltammetric indirect determination of Al(III) by the catalytic cathodic response of norepinephrine at the hanging mercury drop electrode. AB - The biological effects of aluminum (Al) have received much attention in recent years. Al is of basic relevance as concern with its reactivity and bioavailability. In this paper, the electrochemical behaviors of norepinephrine (NE) in the absence and presence of Al(III) at the hanging mercury drop electrode have been studied and applied to the practical analysis. Highly selective catalytic cathodic peak of NE is yielded by linear scan voltammetry (LSV) at 1.32 V (vs. SCE). A linear relationship holds between the cathodic peak current and the Al(III) concentration. It has been successfully applied to the determination of Al(III) in real waters and synthetic biological samples with satisfying results, which are in accordance with those obtained by ICP-AES method. The electrochemical properties and the mechanisms of the peaks in the presence and absence of Al(III) have been explored. The results show that they are irreversible adsorptive hydrogen catalytic waves. These studies not only enrich the methods of determining Al, but also lay foundations of further understanding of the mechanisms of neurodementia. PMID- 16039720 TI - Copper(II) complexes of tridentate pyridylmethylethylenediamines: role of ligand steric hindrance on DNA binding and cleavage. AB - Copper(II) complexes of three linear unsymmetrical tridentate ligands viz. N methyl-N'-(pyrid-2-ylmethyl)ethylenediamine (L1), N,N-dimethyl-N'-(pyrid-2 ylmethyl)ethylenediamine (L2) and N,N-dimethyl-N'-((6-methyl)pyrid-2 ylmethyl)ethylenediamine (L3) have been isolated and characterized by elemental analysis, electronic absorption and EPR spectroscopy and cyclic and differential pulse voltammetry. Of these complexes [Cu(L2)Cl2] and [Cu(L3)Cl2] have been structurally characterized by X-ray crystallography. The [Cu(L2)Cl2] complex crystallizes in the monoclinic space group P2(1)/n with a=11.566(2) A, b=7.369(1) A, c=15.703(3) A, alpha=90 degrees , beta=109.68(8) degrees , gamma=90 degrees and Z=4 while [Cu(L3)Cl2] crystallizes in the triclinic space group P1 with a=9.191(2) A, b=12.359(3) A, c=14.880(3) A, alpha=79.61(13) degrees , beta=86.64(13) degrees , gamma=87.28(8) degrees and Z=2. The coordination geometries around copper (II) in these two complexes are best described as trigonal bipyramidal distorted square based pyramidal (TBDSBP). The distorted CuN3Cl basal plane in them is comprised of three nitrogen atoms of the meridionally coordinated ligand and a chloride ion and the axial position is occupied by the other chloride ion. The interaction of these complexes with Calf Thymus DNA (CT DNA) has been studied by using absorption, emission and circular dichroic spectral methods, thermal denaturation studies, viscometry and cyclic and differential pulse voltammetry. A strong blueshift in the ligand field band and a redshift in the ligand based bands of the copper(II) complexes on binding to DNA imply a covalent mode of DNA binding of the complexes, which involves coordination of most possibly guanine N7 nitrogen of DNA to form a CuN4 chromophore. This is supported by studying the interaction of the complexes with N-methylimidazole (N-meim), guanosine monophosphate (GMP), adenosine monophosphate (AMP) and cytidine (cytd) by ligand field and EPR spectral methods, which indicate the formation of a CuN4 chromophore only in the case of the more basic N-meim and GMP. The DNA melting curves obtained in the presence of copper(II) complexes reveal a monophasic and irreversible melting of the DNA strands and the high positive DeltaTm values (12-21 degrees C) also support the formation of strong Cu-N bonds by the complexes with DNA, leading to intra- and/or interstrand crosslinking of DNA. Competitive ethidium bromide (EthBr) binding studies show that the L2 and L3 complexes are less efficient than the L1 complex in quenching EthBr emission, which is consistent with their forming DNA crosslinking preventing the displacement of the DNA-bound EthBr. A very slight decrease in relative viscosity of DNA is observed on treating the L1 and L2 complexes with CT DNA; however, a relatively significant decrease is observed for the L3 complex suggesting that the length of the DNA fiber is shortened. DNA cleavage experiments show that all the complexes induce the cleavage of pBR322 plasmid DNA, the complex of L1 being more efficient than those of sterically hindered L2 and L3 ligands. PMID- 16039721 TI - Fos expression in the vestibular brainstem: what one marker can tell us about the network. AB - Fos inducible transcription factor expression in rodent brains (rats and gerbils) during manipulations of vestibular input is reviewed. Stimuli included centripetal hypergravity, unilateral labyrinth lesion or semicircular canal plugging, rotational axis cross-coupling (Coriolis forces), high and low rotational vestibulo-ocular reflex gain adaptation, translabyrinth galvanic stimulation, pharmacological manipulation, and combinations thereof. Each type of stimulation elicited unique but partially redundant response patterns in the vestibulo-olivo-cerebellar (VOC) network that reflect the origin and interaction of the labyrinth inputs. On the basis of these patterns, a trained observer can predict what the animal experienced during testing; the patterns of VOC Fos expression reveal a trace of recent genomic activity. Based on principal component analysis, VOC network modules associated with lesion recovery, spatial representation and the calibration of gravity, and optokinetic influences are proposed. Probable and possible gene targets of the Fos protein are also reviewed. PMID- 16039722 TI - Reflexes evoked by electrical stimulation of afferent axons in the pudendal nerve under empty and distended bladder conditions in urethane-anesthetized rats. AB - This study examined reflex mechanisms that mediate urinary bladder and external urethral sphincter (EUS) coordination in female Sprague-Dawley urethane anesthetized rats under empty and distended bladder conditions. The bladder was distended either by a small balloon or a saline filled catheter inserted through the body of the bladder. Stimulation of the entire pudendal nerve elicited short latency (8-12 ms) responses in the EUS and short (3-8 ms) and long latency responses (16-20 ms) in contralateral pudendal nerve. The long latency pudendal pudendal reflex was reduced by 36.7% in area during bladder distension with the balloon catheter. However, there was no significant change in the area of pudendal-EUS reflex during bladder distension. Peak amplitudes of both reflexes were reduced 32% by bladder distension. The effects of glutamatergic receptor antagonists on the reflexes were also examined. MK 801 (0.3-5mg/kg, i.v.), an N methyl-d-aspartate glutamatergic receptor antagonist, markedly depressed the pudendal-pudendal reflex, but LY 215490 (3mg/kg, i.v.), an alpha-amino-5-methyl isoxazole-4-propionate antagonist, had a minimal inhibitory effect. Both glutamatergic receptor antagonists significantly suppressed the pudendal-EUS reflex. These results indicate that the EUS is innervated by multiple pathways and that glutamatergic excitatory transmission is important in the neural mechanisms underlying bladder-sphincter coordination in the rat. PMID- 16039723 TI - Characterization of BF2 and beta2m in three Chinese chicken lines. AB - Twenty-four BF2 genes and 10 beta(2)m genes from Chinese Sanhuang (SH), Wuji (WJ), and Zhenzhu (ZZ) chicken lines were cloned, and the amino acid replacement rates of the BF2 polypeptide binding domain were investigated. For this purpose, 13 BF2 genes from the SH-chicken line (BF2*01sh-BF2*13sh), six BF2 genes from the WJ-chicken line (BF2*01wj-BF2*06wj), and five BF2 genes from the ZZ-chicken line (BF2*01zz-BF2*05zz) were analyzed. The overall conservation of BF2 alleles could be observed within the sequences, and relative conservation was also displayed in the peptide-binding domain, CD8(+) interaction sites, and beta(2)m contact sites. Based on the amino acid similarity, BF2 from the three chicken lines could be divided into eleven gene groups, and five novel gene groups were observed. Although the amino acid similarity among the different alleles was 75.7-99.2%, within an allelic group the members shared >91% amino acid identity with each other. In addition, beta(2)m genes from the three Chinese chicken lines were also clustered into two gene groups: I and II. Between groups I and II, the amino acid identical ratio was much lower (81.9-84.0%). Group I is close to that of the reported chicken beta(2)m, whereas group II represents a new allelic group. The results suggest that five new BF2 groups and a new beta(2)m group exist in the three Chinese chicken lines. PMID- 16039724 TI - Skin immune responses in cattle after primary and secondary infections with Hypoderma lineatum (Diptera: Oestridae) larvae. AB - Cellular immune responses were examined in the skin of cattle after primary and secondary experimental infections with Hypoderma lineatum larvae. Skin biopsies were taken at 0, 6, 12, 48 and 96 h post-infection (h.p.i.). In primary infected animals the penetration of Hypoderma larvae was characterized by moderate inflammatory responses. The pattern of cellular changes in previously infected animals suggested an allergic or rheumatic process, probably as a consequence of the development of a type III hypersensitivity reaction against the second or successive reinfections. Perivascular infiltration with CD3(+) T lymphocytes was marked in infected groups, but especially in previously infected animals, with a significant increase with respect to uninfected controls at 48 h.p.i. B cells remained close to control values during primary infection, while they increased significantly 12 h.p.i. in reinfected animals. IgG(+) plasma cells were also very abundant during secondary infections, with significant differences from primary infected and uninfected animals from 6 h.p.i. onwards. These results suggest that secondary infections allow cellular responses which may be effective in killing some of the entering larvae, resulting in a degree of resistance. PMID- 16039725 TI - Retinal degeneration in experimental coronavirus retinopathy (ECOR) is associated with increased TNF-alpha, soluble TNFR2 and altered TNF-alpha signaling. AB - Experimental coronavirus retinopathy (ECOR) is a virally triggered model of retinal degeneration composed of both genetic and autoimmune components. Since TNF-alpha plays a role in immune-mediated processes we evaluated the levels of TNF-alpha/TNF-alpha receptors and the downstream signaling molecule nitric oxide (NO) during disease in both retinal degeneration susceptible BALB/c and degeneration resistant CD-1 mice. Following coronavirus injection, TNF-alpha mRNA was detected at higher levels within the retinas, and concentrations of TNF-alpha (p<0.005) and sTNFR1 (p<0.0005) proteins were increased within the sera of BALB/c but not CD-1 mice. While concentrations of sTNFR2 proteins were elevated in both BALB/c (p<0.00005) and CD-1 (p<0.005) mice compared to controls, concentrations were higher in BALB/c mice (p<0.0005). Gene expression of iNOS while initially high in BALB/c mice decreased during the acute phase of infection, while it increased in CD-1 mice. These trends are attributable to differences in monocyte TNFR2 release (p<0.0005) between the strains since sTNFR2 decreased (p<0.01) levels of NO production. These studies demonstrate that retinal degeneration following viral infection is associated with increased release of TNF-alpha/TNF receptors combined with a down-regulation of NO. Furthermore they suggest that these molecules are involved in alterations in immune response leading to autoimmune reactivity. PMID- 16039726 TI - Myofunctional evaluation after surgery for tonsils hypertrophy and its correlation to breathing pattern: a 2-year-follow up. AB - OBJECTIVES: To evaluate the myofunctional status in children submitted to adenoidectomy or adenotonsillectomy, correlating the pre and post-surgical patterns throughout a 24-month-period. To correlate the myofunctional alterations to the sort of surgery performed (adenoidectomy versus adenotonsillectomy) and to the predominant post-surgical breathing pattern (predominantly nasal versus allergic rhinitis). METHODS: Forty children were assessed by the otorhinolaryngologist and speech therapist before and 1-24 months after surgery. In order to evaluate myofunctional status, a pre-structured protocol was designed, which included observations regarding facial posture, tonicity, mastication, deglutition and respiration. A score from 0 (normal pattern) to 12 (highly altered) was set. RESULTS: There was a partial, but progressive decrease of the score after surgery (p < 0.001). This decrease was markedly observed during the first 6 months following surgical procedure (p < 0.001), after which it was no longer significant. There was no correlation between the myofunctional progress and the sort of surgery performed. The myofunctional improvement was more accentuated in nasal breathers when compared to those with allergic rhinitis. CONCLUSIONS: Improvement of myofunctional status seems to be observed in children after surgery. In this study, the improvement was predominantly accomplished during the first 6 months following surgical procedure. Persistent pattern of mouth breathing due to allergic rhinitis may difficult recovery of the myofunctional status. The most adequate post-surgical moment for the otorhinolaryngologist to refer the patient to speech therapist for myofunctional therapy seems to be crucial, as well as the recognition by the speech therapist of the persistence of the obstructive symptoms, re-referring this patient to the physician. PMID- 16039727 TI - Inactivation of the rat dorsal striatum impairs performance in spatial tasks and alters hippocampal theta in the freely moving rat. AB - We analysed the interaction between the dorsal striatum (motor coordination and planning) and the hippocampus (sensory information processing and integration) during performance of goal-directed tasks. The performance of rats that had been injected with different doses of the D(2)-antagonist Sulpiride into the dorsal striatum was tested in an egocentric 4-arm maze task that tests striatal functions. Furthermore, hippocampal EEGs were recorded before, during and after inactivation of the dorsal striatum via injections of Sulpiride of rats that were performing a continuous alternation task. Injection of 5 microl of 100 mM Sulpiride increased the number of errors committed in the egocentric 4-arm maze (p < 0.01), indicating that the dorsal striatum is involved in motor control and motor memory recall in such a task. In the recording study, the same dose of Sulpiride injected into the dorsal striatum had powerful effects on the hippocampal EEG. The main activity in the theta range (5-10 Hz) was shifted from higher frequencies in the 8-10 Hz range to lower frequencies in the 5-7 Hz range (p < 0.005). The impairment in the behavioural egocentric task after Sulpiride injection, and the effects of Sulpiride on hippocampal theta shows that there is a functional interaction between the dorsal striatum and the hippocampus. While the dorsal striatum coordinates the execution of complex motor programs, the hippocampus integrates spatial and other sensory information required for the planning and execution of goal-directed movements. PMID- 16039728 TI - The expression of prepulse inhibition of the acoustic startle reflex as a function of three pulse stimulus intensities, three prepulse stimulus intensities, and three levels of startle responsiveness in C57BL6/J mice. AB - Using the acoustic startle reflex, prepulse inhibition (PPI) is typically demonstrated by the interaction between two auditory stimuli presented in close temporal proximity. When a startle-eliciting pulse stimulus is shortly preceded by a weak prepulse stimulus, the reaction to the former is attenuated in comparison to when the pulse stimulus is presented alone. The present experiment evaluated the influence of different prepulse and pulse intensities upon the expression of PPI by additionally taking into account individual differences in startle reactivity. To this end, we subdivided a cohort of 102 mice into three subsets of equal size differing in startle responsiveness, and evaluated PPI using three levels of prepulse stimulus in combination with three intensities of pulse stimulus. Our results revealed additive as well as complex interactive effects amongst individual's reactivity, pulse intensity and prepulse intensity. At the same time, additional issues concerning the quantification of PPI are highlighted, especially when startle reactivity differs considerably between comparison groups. We concluded that the variation in pulse intensity represents a valuable addition to PPI assessment in general, and especially in genetically modified mice. PMID- 16039729 TI - Neuro-apoptogenic and blood platelet targeting toxins in benthic marine cyanobacteria from the Portuguese coast. AB - Six strains of marine cyanobacteria, of which five benthic, were isolated from an area of the Portuguese coast with no known apparent toxic microbial bloom. Five strains were lethal for mice. Four of them produced lethargy and four lead to bleeding. One of the toxic strains was from a genus (Aphanothece) not previously associated with toxin production. Extracts from four isolates induced SH-SY5Y neuroblastoma cell apoptosis without affecting the viability of hepatocytes, NRK kidney cells, or fibroblasts. Aqueous extract from four isolates inhibited thrombin-induced blood platelet activation, with decreased P-selectin expression, platelet aggregation and shedding of platelet-derived micro-vesicles. Curiously, platelets treated with organic extracts from two of the cyanobacterial strains formed platelet micro-vesicles, expressed P-selectin on the surface and showed a distinct phosphotyrosine protein pattern, but failed to aggregate. We conclude that low-abundance marine cyanobacteria growing at low rates may be an important source for novel toxins that may be useful to dissect mammalian signalling pathways of apoptosis and platelet function. PMID- 16039730 TI - Ras signaling from plasma membrane and endomembrane microdomains. AB - Ras proteins are compartmentalized by dynamic interactions with both plasma membrane microdomains and intracellular membranes. The mechanisms underlying Ras compartmentalization involve a series of protein/lipid, lipid/lipid and cytoskeleton interactions, resulting in the generation of discrete microdomains from which Ras operates. Segregation of Ras proteins to these different platforms regulates the formation of Ras signaling complexes and the generation of discrete signal outputs. This temporal and spatial modulation of Ras signal transduction provides a mechanism for the generation of different biological outcomes from different Ras isoforms, as well as flexibility in the signal output from a single activated isoform. PMID- 16039731 TI - Prevalence of inducible paroxysmal supraventricular tachycardia during esophageal electrophysiologic study in patients with unexplained stroke. AB - BACKGROUND: The relationships between stroke and atrial tachycardia or atrial fibrillation were previously reported. Electrophysiological study is one of the means, used to detect and evaluate these atrial tachyarrhythmias. But, some other arrhythmias as paroxysmal supraventricular tachycardia, can be induced during electrophysiologic study and their significance in stroke is unknown. The aim of the study was to assess the significance of inducible paroxysmal supraventricular tachycardia (PSVT) in stroke. METHODS: One hundred thirty seven patients, aged 61+/-12 years had unexplained stroke (group I) and were compared to 60 subjects aged 45+/-18.5 years without stroke and history of tachycardia (group II); Holter monitoring (HM), echocardiogram and esophageal electrophysiologic study (EPS) in basal state and after isoproterenol were performed. RESULTS: Heart disease was noted in 19 group I patients (14%) and 10 group II patients (17%). In group I, atrial fibrillation or tachycardia (AF-AT) was induced in 20 patients (15%) and PSVT was induced in 19 patients (14%) aged 66+/-12 years. In group II, AF/AT was induced in 3 patients (5%); no group II patient had induced PSVT. After 3+/-1 years, in group I, one of 98 patients without induced arrhythmias had new strokes and 2 had AF; 5 patients with induced AT/AF developed AF; 5 patients with induced PSVT had PSVT's, requiring ablation in 4 of them; 1 died from a new stroke; one had a second non-fatal stroke and 3 patients developed AF (16%). In group II, there were no events. CONCLUSION: In 14% of patients with unexplained stroke, PSVT was inducible during esophageal electrophysiologic study. Further studies are warranted to assess the significance of this finding in patients with unexplained stroke. PMID- 16039732 TI - Coarctation of the aorta evaluated with 64-row multislice computed tomography. PMID- 16039733 TI - Comparison of N-acetylcysteine and fenoldopam for preventing contrast-induced nephropathy (CAFCIN). AB - BACKGROUND: N-acetylcysteine and fenoldopam are commonly prescribed for prevention of contrast mediated nephropathy, however, comparative superiority of either agent is unknown. METHODS: In a prospective, randomized, parallel-group trial, adult cardiac catheterization patients at the university and veterans' hospitals with pre-existing stable renal insufficiency were randomized to N acetylcysteine 600 mg orally twice daily for 4 doses or fenoldopam 0.1 mcg/kg/min intravenously for a minimum of 8 h. All patients received intravenous hydration with normal saline (5% dextrose in normal saline for diabetics on insulin). Randomization was stratified for diabetes. The primary endpoint was mean change in Scr at 72 h. Secondary endpoint was the incidence of contrast-induced nephropathy (25% increase above baseline Scr or absolute increase of 0.5 mg/dL). RESULTS: Study termination occurred after ninety-five patients (mean age 68+/-10 years, female 25%, diabetic 42%, mean baseline Scr 1.5+/-0.4 mg/dL) were randomized, with 84 completing follow-up (44 N-acetylcysteine, 40 fenoldopam). Overall, there were no significant differences in mean change in Scr at 72 h (N acetylcysteine 0.20+/-0.72 vs. fenoldopam 0.08+/-0.48 mg/dL, p=0.4) or incidence of contrast-induced nephropathy (N-acetylcysteine 5 vs fenoldopam 8, p=0.4). No differences were detected in subgroup analyses for diabetes, baseline Scr >1.7 or 2.0 mg/dL, gender, age >70 years, or contrast volume >150 mL. Results were similar after multivariate adjustment for diabetes, contrast volume, heart failure and gender. CONCLUSIONS: Our randomized comparison failed to demonstrate a significant difference in the abilities of N-acetylcysteine and fenoldopam to prevent the decline in renal function or the incidence of contrast-induced nephropathy during cardiac catheterization. PMID- 16039734 TI - Procainamide-induced pure red cell aplasia. PMID- 16039735 TI - Are predictors for myocardial infarction the same for women and men when evaluated prior to hospital admission? AB - AIM: To describe predictors of myocardial infarction prior to hospital admission in women and men among patients with a suspected acute coronary syndrome without ST-elevation. DESIGN: Prospective observational study in Stockholm and Goteborg, Sweden. RESULTS: Of 433 patients who did fulfill the inclusion criteria 45% were women. Fewer women (17%) than men (26%) developed acute myocardial infarction (AMI) (p=0.054), particularly among patients with initial ST-depression, in whom AMI was developed in 22% of women and 54% of men (p = 0.001). Predictors for infarct development in women were: a history of AMI and advanced age. Among men they were: initial ST-depression or a Q-wave on ECG and elevation of biochemical markers (both recorded on admission of the ambulance crew). There was a significant interaction between gender and the influence of ST-depression on the risk for development of myocardial infarction (p < 0.05). CONCLUSION: Among patients transported with ambulance due to a suspected acute coronary syndrome and no ST-elevation fewer women than men seem to develop AMI particularly among patients with ST-depression. These results suggest that early prediction of myocardial infarction might differ between women and men with acute chest pain. PMID- 16039736 TI - The treadmill effect in a fixed budget system. AB - We examine the interaction in the market for physician services when the total budget for reimbursement is fixed. Physicians obtain points for the services they render. At the end of the period the budget is divided by the sum of all points submitted, which determines the price per point. We show that this retrospective payment system involves -- compared to a fee-for-service remuneration system -- a severe coordination problem, which potentially leads to the "treadmill effect". We argue that when market entry is possible, a budget can be efficiency enhancing, if in addition a price floor is used. PMID- 16039737 TI - The effect of preconditioning on the Cu, Zn superoxide dismutase expression and enzyme activity in rat brain at the early period after severe hypobaric hypoxia. AB - Severe hypoxia results in functional and structural injury of the brain. A preconditioning with repetitive episodes of mild hypoxia considerably ameliorates neuronal resistance to subsequent severe hypoxia. Activation of endogenous antioxidants including Cu, Zn-depending superoxide dismutase (Cu, Zn-SOD) (EC.1.15.1.1) is one of the main cell defense mechanisms against oxidative stress induced by hypoxia. Alterations of expression and enzyme activity of Cu, Zn-SOD 3 and 24h after severe hypobaric hypoxia in forebrain structures of preconditioned and non-preconditioned rats were investigated. We found that hypoxia without preconditioning suppressed the Cu, Zn-SOD enzyme activity at 3h time-point but preconditioning essentially modified the reaction to severe hypoxia by increasing the expression and activity of Cu, Zn-SOD during early stages of reoxygenation crucial for apoptosis initiation. PMID- 16039738 TI - Enhanced cardiovascular alteration and Fos expression induced by central salt loading in a conscious rat transgenic for the metallothionein-vasopressin fusion gene. AB - The present study is an investigation of the responses of the cardiovascular system and Fos expression to intracerebroventricular (i.c.v.) administration of hypertonic saline (HS) in conscious arginine vasopressin (AVP)-overexpressing transgenic (Tg) and control rats. Central HS (0.3, 0.67, or 1.0M NaCl, 1 microl/min for 20 min) significantly increased the mean arterial blood pressure (MABP) and Fos-like immunoreactivity (FLI) in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) of the hypothalamus, the area postrema (AP), the median preoptic nucleus (MnPO), and the organum vasculosum laminae terminalis (OVLT) in both Tg and control rats. The changes in MABP and FLI were significantly larger in Tg rats than in control rats. i.c.v. pretreatment with the AVP V1 receptor antagonist, OPC-21268, blocked the increase in MABP and significantly decreased the Fos expression in the PVN (posterior magnocellular (pm) component) induced by 0.3 M HS in the Tg rats. The present study demonstrates an increased responsiveness to i.c.v. administration of HS in AVP Tg rats, suggesting the relationship between the vasopressinergic drive and central cardiovascular response via, at least in part, the V1 receptor in the PVN magnocellular neurons. PMID- 16039739 TI - Diurnal fluctuation of time perception under 30-h sustained wakefulness. AB - Previous studies have reported that time perception in humans fluctuates over a 24-h period. Behavioral changes seem to affect human time perception, so that the fluctuation in human time perception may be the result of such changes due to self-determined activities. Recently, we carried out a study in which a healthy human cohort was asked to perform simultaneously loaded cognitive tasks under controlled conditions, and found that time perception decreased linearly from morning to evening. In addition, the variations in time perception were not a consequence of behavioral changes. It remains to be elucidated whether diurnal variations in time perception are a consequence of circadian rhythm or of some homeostatic changes that are attributable to accumulated wake time. The effects of circadian rhythm on time perception were investigated in eight healthy young male volunteers by conducting 10-s time production tasks under 30-h constant routine conditions. Core body temperature and serum melatonin and cortisol levels were measured during the course of the study. Produced time exhibited a diurnal variation and was strongly correlated with circadian variations in core body temperature and serum melatonin levels. These results suggest that human short term time perception is under the influence of the circadian pacemaker. PMID- 16039740 TI - Interaction between paired-pulse facilitation and long-term potentiation in the projection from hippocampal area CA1 to the entorhinal cortex. AB - The hippocampus communicates with the neocortex via the entorhinal cortex and is thought to be critically involved in the consolidation of memories. This paper contains in vivo evidence of a projection from the hippocampal area CA1 to the entorhinal cortex. Current theories of memory formation suggest that the backprojections from the hippocampus to the neocortex should undergo some form of plastic change in order that memories become consolidated. Paired-pulse facilitation (PPF) and long-term potentiation (LTP) are forms of short- and long term plasticity, respectively. We show that the CA1 to entorhinal cortex projection is capable of sustaining PPF over a wide range of stimulus intervals. In addition we demonstrate that following high frequency stimulation of this pathway the evoked response in the entorhinal cortex remains potentiated for at least 30 min. Finally, we demonstrate that PPF changes following LTP depending on the initial ratio of PPF, suggesting that LTP expression on this pathway may contain a presynaptic component. These findings should provide insight into the hippocampal function in memory formation. PMID- 16039741 TI - First ICFMH workshop: Food Safety in Africa, University of Stellenbosch, Stellenbosch, South Africa, 8-13 December 2003. PMID- 16039742 TI - Enterobacter sakazakii: a coliform of increased concern to infant health. AB - The first cases of neonatal meningitis believed to have been caused by Enterobacter sakazakii were reported in 1961. Prompted by several subsequent outbreaks of E. sakazakii infections in neonates and an increasing number of neonates in intensive care units being fed rehydrated powdered infant formula, considered to be a source of the pathogen, public health authorities and researchers are exploring ways to eliminate the bacterium or control its growth in dry infant formula, processing environments and formula preparation areas in hospitals. Reviewed here are advances in taxonomy and classification of E. sakazakii, methods of detecting, isolating and typing the bacterium, antibiotic resistance, clinical etiology and pathogenicity. Outbreaks of E. sakazakii infections in neonates and adults are summarized. Reports on the presence of E. sakazakii in clinical settings, the environment and foods and food processing facilities are reviewed. Tolerance of the pathogen to environmental stresses, its behavior in powdered and rehydrated infant formulae and hazard analysis and risk management are discussed. Research needs are presented. PMID- 16039743 TI - Heterologous production of five Hepatitis C virus-derived antigens in three Saccharomyces cerevisiae host strains. AB - In this study, the production of recombinant Hepatitis C virus (HCV) derived proteins from transformed Saccharomyces cerevisiae yeast cells is reported. Three different yeast strains (GRF18U, BY4743-4A and CENPK 113-5D) have been transformed for the intracellular expression of five antigens of different dimensions (from 32.8 to 85.2 kDa), all derived from the non-structural (NS) region of different HCV viruses' genotypes and posed under the control of a glycolytic promoter. The putative trans-membrane domains contained in four antigens seem responsible of their accumulation as protein aggregates. Good productions of the smaller and of the bigger antigens (50 and 30 mgl(-1), respectively) have been observed in simple flask batch cultures. Productions are strongly dependent from the genetic background of the yeast host and from the cellular localization of the antigen, while they appear independent from the growth rate of the transformed hosts. For every recombinant antigen tested, the highest production levels were achieved with the CENPK 113-5D-host strain, while the GRF18U strain shows symptoms of a heavily stressed phenotype. PMID- 16039744 TI - Enzyme stabilization by glutaraldehyde crosslinking of adsorbed proteins on aminated supports. AB - The stabilization achieved by different immobilization protocols have been compared using three different enzymes (glutaryl acylase (GAC), D-aminoacid oxidase (DAAO), and glucose oxidase (GOX)): adsorption on aminated supports, treatment of this adsorbed enzymes with glutaraldehyde, and immobilization on glutaraldehyde pre-activated supports. In all cases, the treatment of adsorbed enzymes on amino-supports with glutaraldehyde yielded the higher stabilizations: in the case of GOX, a stabilization over 400-fold was achieved. After this treatment, the enzymes could no longer be desorbed from the supports using high ionic strength (suggesting the support-protein reaction). Modification of the enzymes immobilized on supports that did not offer the possibility of react with glutaraldehyde showed the same stability that the non modified preparations demonstrating that the mere chemical modification did not have effect on the enzyme stability. This simple strategy seems to permit very good results in terms of immobilization rate and stability, offering some advantages when compared to the immobilization on glutaraldehyde pre-activated supports. PMID- 16039746 TI - An enteric-coated dry emulsion formulation for oral insulin delivery. AB - A novel oral dosage formulation of insulin consisting of a surfactant, a vegetable oil, and a pH-responsive polymer has been developed. First, a solid-in oil (S/O) suspension containing a surfactant-insulin complex was prepared. Solid in-oil-in-water (S/O/W) emulsions were obtained by homogenizing the S/O suspension and the aqueous solution of hydroxypropylmethylcellulose phthalate (HPMCP). A microparticulate solid emulsion formulation was successfully prepared from the S/O/W emulsions by extruding them to an acidic aqueous solution, followed by lyophilization. The insulin release from the resultant dry emulsion responded to the change in external environment simulated by gastrointestinal conditions, suggesting that the new enteric-coated dry emulsion formulation is potentially applicable for the oral delivery of peptide and protein drugs. PMID- 16039745 TI - Phylogenetic analyses of type A influenza genes in natural reservoir species in North America reveals genetic variation. AB - The matrix (M) (98 isolates), nucleoprotein (NP) (67 isolates), non-structural (NS) (107 isolates), H4 subtype HA1 (21 isolates) and H6 (38 isolates) subtype HA1 region of the hemagglutinin (HA) gene were sequenced from avian influenza viruses isolated from North American wild aquatic birds between 1969 and 2003. Sequences were compared phylogenetically with all available wild aquatic bird isolate sequences and sequences from isolates from gallinaceous poultry and mammalian host species. Among the wild aquatic bird isolates the NS subtype B NS1 and subtype B NS2 proteins were the most conserved with minimum amino acid identities of 98.0 and 99.0%, respectively. The H6 HA1, M and NS subtype A genes were more divergent; both nucleotide and amino acid divergence levels were similar to those of the same genes from gallinaceous poultry and mammalian origin type A influenza isolates. Phylogenetically none of the genes assorted geographically (within North America), chronologically or by species of origin, unlike isolates from non-natural host species, suggesting that selection pressure is low. A lack of clear temporal or spatial grouping shows that multiple lineages of virus co-circulate and precludes the development of an epidemiological map for influenza virus from these species. PMID- 16039747 TI - In vivo tumor transfection mediated by polyplexes based on biodegradable poly(DMAEA)-phosphazene. AB - In recent years, increasing interest is being paid to the design of transfectants based on non-toxic and biodegradable polymers for gene therapy purposes. We recently reported on a novel, biodegradable polymer, poly(2-dimethylamino ethylamino)phosphazene (p(DMAEA)-ppz) for use in non-viral gene delivery. In this study, the biodistribution and in vivo transfection efficiency of polyplexes composed of plasmid DNA and p(DMAEA)-ppz were investigated after intravenous administration in tumor bearing mice. Data were compared with those of polyplexes based on the non-biodegradable polyethylenimine (PEI 22kDa). Both polyplex systems were rapidly cleared from the circulation (<7% ID, at 60 min after administration) and showed considerable disposition in the liver and the lung, all in line with earlier work on cationic polyplex systems. The lung disposition is attributed to aggregates formed by interaction of the polyplexes with blood constituents. Redistribution of the polyplexes from the lung was observed for both polyplex formulations. Importantly, both polyplex systems showed a substantial tumor accumulation of 5% and 8% ID/g for p(DMAEA)-ppz and PEI22 polyplexes, respectively, at 240 min after administration. The tumor disposition of the p(DMAEA)-ppz and PEI22 polyplexes was associated with considerable expression levels of the reporter gene. In contrast to PEI22 polyplexes, p(DMAEA) ppz polyplexes did not display substantial gene expression in the lung or other organs (organ gene expression<1/100 of tumor gene expression). The observed preferential tumor gene expression mediated by the p(DMAEA)-ppz polyplexes enables the application of this polymer to deliver therapeutic genes to tumors. PMID- 16039748 TI - Impaired expression and function of the bile salt export pump due to three novel ABCB11 mutations in intrahepatic cholestasis. AB - BACKGROUND/AIMS: Inherited dysfunction of the bile salt export pump BSEP (ABCB11) causes a progressive and a benign form of familial intrahepatic cholestasis, denominated as PFIC2 and BRIC2, respectively. We functionally characterized novel ABCB11 mutations encountered in two patients with a PFIC2 and a BRIC2 phenotype, respectively. METHODS: BSEP expression was determined in liver biopsies by immunohistochemistry. ABCB11 mutations were functionally characterized by taurocholate transport in SF9 cells transfected with human ABCB11. RESULTS: The PFIC2 patient was compound heterozygous for a splicing mutation in intron 4 ((+3)A > C) combined with an early stop codon at position 930 (R930X), while the BRIC2 patient was compound heterozygous for two nonsynonymous mutations in exon 9 (E297G) and exon 12 (R432T), respectively. Hepatic BSEP expression was absent in PFIC2 and preserved in BRIC2. In BRIC2, taurocholate transport was decreased to 13% and 20% of reference levels for R432T and E297G, respectively. CONCLUSIONS: The intron 4 (+3)A > C, R930X and R432T represent previously undescribed mutations of the ABCB11 gene that confer a PFIC2 and a BRIC2 phenotype, respectively. By combining functional in-vitro characterization with immunohistochemical detection of variant BSEP we provide direct evidence for the role of ABCB11 mutations in the pathogenesis of different forms of intrahepatic cholestasis. PMID- 16039749 TI - Chicken orthologues of mammalian imprinted genes are clustered on macrochromosomes and replicate asynchronously. AB - In the chicken genome, most orthologues of mouse imprinted genes are clustered on macrochromosomes. Only a few orthologues are located in the microchromosome complement. Macrochromosomal and, to a lesser extent, microchromosomal regions containing imprinted gene orthologues exhibit asynchronous DNA replication. We conclude that highly conserved arrays of imprinted gene orthologues were selected during vertebrate evolution, long before these genes were recruited for parent specific gene expression by genomic imprinting mechanisms. Evidently, the macrochromosome complement provides a better chromatin environment for the establishment of asynchronous DNA replication and imprinted gene expression later in evolution than microchromosomes. PMID- 16039750 TI - Efficient implementation of non-linear valve law and ventricular interaction dynamics in the minimal cardiac model. AB - A minimal model of the cardio-vascular system (CVS) with ventricular interaction and inertial effects that accurately captures the physiological trends of a variety of disease states has been developed. However, the physiologically accurate open on pressure, close on flow valve law is computationally heavy to implement, reducing the model's potential clinical benefit. A significantly simpler representation of the valve law using Heaviside functions is derived and the ventricular interaction equations are reformulated to obtain a unique closed form analytical solution. The new formulation is tested and compared with the previous formulation for a healthy human and four clinically significant disease states: mitral and aortic stenosis, pulmonary embolism and septic shock. The new model formulation matches the previous model definition, differing by a mean model response error of no more than 0.2%. Computationally, it is 24 x faster than the previous method. More specifically, a short 20-beat simulation that took 102 s now requires 4.3 s, significantly improving the model's potential for practical use in a diagnostic and/or decision support role in the intensive care unit. PMID- 16039751 TI - Numerical simulations of pyrite oxidation and acid mine drainage in unsaturated waste rock piles. AB - Numerical simulations of layered, sulphide-bearing unsaturated waste rock piles are presented to illustrate the effect of coupled processes on the generation of acid mine drainage (AMD). The conceptual 2D systems were simulated using the HYDRUS model for flow and the POLYMIN model for reactive transport. The simulations generated low-pH AMD which was buffered by sequential mineral dissolution and precipitation. Sulphide oxidation rates throughout the pile varied by about two orders of magnitude (0.004-0.4 kg m-3 year-1) due to small changes in moisture content and grain size. In the fine-grained layers, the high reactive surface area induced high oxidation rates, even though capillary forces kept the local moisture content relatively high. In waste rock piles with horizontal layers, most of the acidity discharged through vertical preferential flow channels while with inclined fine grained layers, capillary diversion channeled the AMD to the outer slope boundary, keeping the pile interior relatively dry. The simulation approach will be useful for helping evaluate design strategies for controlling AMD from waste rock. PMID- 16039752 TI - Endogenous opiates and behavior: 2004. AB - This paper is the 27th consecutive installment of the annual review of research concerning the endogenous opioid system, now spanning over 30 years of research. It summarizes papers published during 2004 that studied the behavioral effects of molecular, pharmacological and genetic manipulation of opioid peptides, opioid receptors, opioid agonists and opioid antagonists. The particular topics that continue to be covered include the molecular-biochemical effects and neurochemical localization studies of endogenous opioids and their receptors related to behavior, and the roles of these opioid peptides and receptors in pain and analgesia; stress and social status; tolerance and dependence; learning and memory; eating and drinking; alcohol and drugs of abuse; sexual activity and hormones, pregnancy, development and endocrinology; mental illness and mood; seizures and neurologic disorders; electrical-related activity and neurophysiology; general activity and locomotion; gastrointestinal, renal and hepatic functions; cardiovascular responses; respiration and thermoregulation; and immunological responses. PMID- 16039754 TI - Mass spectrometric assignment of Leu/Ile in neuropeptides from single neurohemal organ preparations of insects. AB - Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF-TOF) tandem mass spectrometry has been applied for the first time on an insect/arthropod target, focusing on PVK/CAP2b neuropeptides in the housefly Musca domestica and flesh fly Neobellieria bullata. The peptidomic analysis of single neurohemal organ preparations allows the unambiguous assignment of internal Leu/Ile positions not distinguishable by previous mass spectrometric techniques. The confirmation of side-chain fragments which allows assignment of Leu/Ile even from samples as small as neurohemal organs will greatly accelerate the identification of novel neuropeptides that are implicated in the regulation of critical physiological processes in insects. The unnatural Ile analog is 4.5 times more active than the native Leu sequence in a housefly Malpighian tubule fluid secretion assay, which reinforces the caveat that potency values in a biological assay cannot be relied upon to predict the native sequence. PMID- 16039755 TI - Ganodermin, an antifungal protein from fruiting bodies of the medicinal mushroom Ganoderma lucidum. AB - A 15-kDa antifungal protein, designated ganodermin, was isolated from the medical mushroom Ganoderma lucidum. The isolation procedure utilized chromatography on DEAE-cellulose, Affi-gel blue gel, CM-Sepharose and Superdex 75. Ganodermin was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel and CM-Sepharose. Ganodermin inhibited the mycelial growth of Botrytis cinerea, Fusarium oxysporum and Physalospora piricola with an IC50 value of 15.2 microM, 12.4 microM and 18.1 microM, respectively. It was devoid of hemagglutinating, deoxyribonuclease, ribonuclease and protease inhibitory activities. PMID- 16039756 TI - The role of lactate in brain metabolism. AB - According to the astrocyte-neurone-lactate shuttle (ANLS) hypothesis, activated neurones use lactate released by astrocytes as their energy substrate. The hypothesis, based largely on in vitro experiments, postulates that lactate is derived from the uptake by astrocytes of synaptically released glutamate. The time course of changes in lactate, derived from in vivo experiments, is incompatible with the ANLS model. Neuronal activation leads to a delayed rise in lactate followed by a slow decay, which greatly outlasts the period of neuronal activation. The present review proposes that the uptake of stimulated glutamate release from astrocytes, rather than synaptically released glutamate, is the source of lactate released following neuronal activation. This rise in lactate occurs too late to provide energy for neuronal activity. Furthermore, there is no evidence that lactate undergoes local oxidative phosphorylation. In conclusion, under physiological conditions, there is no evidence that lactate is a significant source of energy for activated neurones. PMID- 16039757 TI - [Isolated muscular calf vein thrombosis: treat or not to treat?]. PMID- 16039758 TI - [Concomitant discovery of non Hodgkin's lymphoma and bone tuberculosis]. PMID- 16039759 TI - Effects of physical activity on food intake. AB - The understanding of the effect of physical activity (PA) on food intake is imperative for considering PA as an additional tool for prevention and treatment of many diseases. Prolonged strenuous PA performed on a regular basis causes an increase in overall energy turnover, and leads either to loss of body weight, or to an increased food intake. When leading to loss of body weight, PA may be used as a therapeutic adjunct in the treatment of obesity. When increasing food intake to compensate for increased energy expenditure and maintaining body weight, PA, by increasing intake of other constituents of food like minerals and vitamins, may contribute to micronutrient deficiency coverage. However, the type of activity, as well as body composition of the individuals engaging in sport, play an important role in food intake regulation and its changes with time. Some studies indicate that short-term exercise does not have the same effect on food intake as long-term exercise and that eventual increase in food intake due to increased PA does not follow the same pattern in obese as in lean individuals. To better understand the relationship between PA on food intake, current findings on this topic will be summarized in this paper. PMID- 16039760 TI - Efficacious, nontoxigenic Bacillus anthracis spore vaccines based on strains expressing mutant variants of lethal toxin components. AB - We reported previously on the development of a Bacillus anthracis vaccine strain expressing high levels of recombinant protective antigen (rPA) [Cohen et al., Infec Immun 2000;68(8):4549-58]. To further explore the potential of the B. anthracis platform, we generated several attenuated strains expressing lethal toxin components PA and LF, which are biologically inactive, yet retain their antigenic properties. A single injection of 5 x 10(7) spores of one of these strains, carrying PA mutation at a site involved in effector translocation (residues 313-314) was shown to resemble wild type PA in inducing production of high levels of anti-PA neutralizing antibodies and producing effective protective immunity for 12 months. Long-term protection and persistence of functional antibody titers was observed after the gradual elimination of spores from guinea pig tissues 3 months after injection and in the measurable absence of bacteria in tissues. The mutant toxin components could, thus be an effective alternatives to their native counterparts when presented to the immune system in context of a live B. anthracis strain. These live vaccine prototypes may serve as a platform for future multi-component vaccines. PMID- 16039761 TI - The application of new techniques to the improved detection of persistently infected cattle after vaccination and contact exposure to foot-and-mouth disease. AB - Detection of antibodies to the non-structural proteins (NSP) of foot-and-mouth disease virus (FMDV) was compared with conventional serological and virological methods and with RT-PCR for the identification of FMDV carrier animals obtained after experimental contact challenge of vaccinated cattle. Transmission from carriers to sentinels was also monitored. Twenty FMDV vaccinated and five unvaccinated cattle were challenged by direct contact with five donor cattle excreting FMDV and monitored until 28 days post challenge-exposure . Twelve vaccinated and three unvaccinated animals were retained up to 24 weeks post exposure to FMDV in order to monitor viral persistence, transmission and antibody responses. In nine vaccinated animals, infection persisted beyond 28 days post exposure, virus being detected more frequently and for longer in oesophagopharyngeal samples from these animals when examined by RT-PCR rather than by virus isolation. Although recovery of FMDV RNA became increasingly sporadic over time, the number of RNA copies detected in positive samples declined only slowly. Two naive sentinel cattle housed with the persistently infected animals between 93 and 168 days after the latter had been challenge exposed to FMDV did not become infected. There were differences in the ability of commercially available serological tests to detect antibodies to FMDV non structural proteins (NSP) in vaccinated and subsequently challenged cattle. Although no single test could identify all of the vaccinated cattle that became persistently infected, the most poorly recognised animals were those with the least evidence of virus replication based on other tests. The potential of the detection of antibodies to the 2B NSP of FMDV for diagnosing persistent FMDV infection was demonstrated. PMID- 16039762 TI - The macro-epidemiology of influenza vaccination in 56 countries, 1997--2003. AB - The WHO Global Agenda on Influenza calls for measuring the progress of national influenza vaccination programs. In response, the Macro-epidemiology of Influenza Vaccination (MIV) Study Group has gathered information on influenza vaccination in 56 countries. During the period 1997--2003, influenza vaccine distribution increased considerably in almost all countries. In 2003, the countries with the highest levels of vaccination (doses distributed/1000 population) were Canada (344), the Republic of Korea (311), the United States (286) and Japan (230). Most countries recommended influenza vaccination for elderly persons and those with high-risk medical conditions, including immuno-compromise. Fewer countries provided public reimbursement for vaccination through national or social health insurance. Higher levels of vaccination were not closely related to higher levels of economic development, but in many instances public reimbursement for vaccination seemed to be associated with greater vaccine use. From 1994 to 2003, the global use of influenza vaccines increased more than two-fold. In 2003, the 56 MIV Study Group countries accounted for approximately 95% of the 292 million doses of influenza vaccine distributed worldwide, and 62% of these doses were distributed within nine vaccine-producing countries in North America, Western Europe, Japan and Australia. However, influenza vaccination was increasing rapidly in many non vaccine-producing countries, and this change has important implications for pandemic vaccination. PMID- 16039763 TI - Perceived HIV stigma in AIDS caregiving dyads. AB - This study examines perceived HIV stigma in AIDS caregiving dyads in the United States, assessing the measurement of and correlates of personal stigma (among care-recipients living with HIV), courtesy stigma (among caregivers), and dyadic stigma. Survey data from 135 dyads in which the caregiver is a midlife or older mother or wife, and the care-recipient is her HIV-infected adult son or husband, are analyzed with individual-level and multilevel regression models. Results indicate that: (1) perceived stigma can be reliably measured among both persons living with HIV (PLH) and caregivers; (2) personal stigma can be distinguished from courtesy stigma; (3) perceived stigma is relatively low in this sample, and is higher among PLH than caregivers, higher among caregiving wives than mothers, and similar between PLH who are husbands and sons; (4) dyadic stigma is influenced by the caregiver's HIV status, the ethnic composition of the dyad, caregiving duration, and household income; (5) stigma discrepancy within dyads is a function of health discrepancy within dyads; and (6) differences in multivariate correlates of perceived stigma at the individual-level, in comparison to the dyad-level, suggest that dyadic stigma is a unique construct. A recognition that perceived stigma bears its own unique influence on the caregiving dyad is important for understanding how best to allocate resources aimed at alleviating stigma among individuals and families impacted by HIV. PMID- 16039764 TI - US-based indices of area-level deprivation: results from HealthCare for Communities. AB - In this analysis we create census-tract level indices of area deprivation for the US that parallel similar indices developed in Britain, and we determine whether these indices are related to physical and mental health outcomes. Data for the indices come from the 2000 Census Summary File Tapes and the 2001 Zip Code Business Patterns Files. These indices are then linked by census tract to cross sectional data from the HealthCare for Communities (HCC) study, and equations are estimated using multi-level models with census-tract random effects. We find that area-level deprivation predicts poor mental and physical health outcomes, but different components of deprivation have different effects. When we measure deprivation using three factor scores that emerged from our analysis (rather than combining all measures of deprivation into a single index), we find that access to services has a more pronounced association with physical health, whereas racial composition and local language barriers are more strongly correlated with mental health. These findings suggest that grouping all variables into a single index may mask important heterogeneity in the ways in which area characteristics affect health outcomes. PMID- 16039765 TI - What accounts for the relationship between social class and smoking cessation? Results of a path analysis. AB - Despite the overall decline in cigarette smoking prevalence in the US, social class inequalities in smoking are likely to persist, or even to widen. One possible reason for the increasing gap in smoking prevalence across social class could be our lack of understanding of causal mechanisms: in other words, what accounts for the social gradient in smoking behavior? In this paper, we examine the mechanisms behind social gradients related to smoking cessation by use of path analysis techniques. The data come from a 3-year follow-up telephone survey of a cohort of US adults. The sample for the present analysis was drawn from the 481 respondents who reported being smokers and employed at baseline and who completed the follow-up interview. We examined two social class indicators, educational attainment and household income, in relation to smoking cessation. We tested the potential mediating effects of the following variables: differential use of resources for smoking cessation (e.g., booklet, pamphlet, quit line, nicotine replacement therapy and smoking cessation program), differential environments in terms of smoking at worksite and home, and differences in peer smoking. Our path analyses suggest that smokers from high social class are likely to use effective resources for smoking cessation and have restrictive home environment in terms of smoking, which leads to a relatively higher smoking cessation rate compared to those from low social class. The results of this study suggest that interventions should target resources for smoking cessation and home environments in terms of smoking to reduce socio-economic disparities in smoking cessation. PMID- 16039766 TI - Contesting contraceptive innovation--reinventing the script. AB - The article describes how the merging of Southern and Northern women's health groups resulted in a powerful transnational movement, with a collective oppositional identity based on shared solidarity in campaigns for reproductive rights and against state coercion in reproductive matters. It focuses on the ways in which the movement framed issues of rights and safety and pointed to the possible abuse potential of two new longer-acting contraceptive technologies, Norplant and the anti-fertility vaccines. The contestations by women's health advocates resulted in the emergence of a strong commitment among scientists to involve women's health advocates in the development and introduction of new contraceptive technologies. By engaging in the construction of safety and efficacy claims, and by outlining conditions for the introduction of the new technologies (so-called introduction scripts) women's health advocates were able to reinscribe the technologies with representations of bodily integrity and reproductive rights, rather than population control. I argue that a split within the women's health movement on the need to ban the new technologies did not weaken its impact, but, in fact, enhanced this success. I describe, in detailed case studies on the Norplant and Anti-fertility vaccine controversies, how both strands of women's health advocacy claim to be able to represent the interest of users, but that their representations of users differ. The 'no-to-Norplant' and 'no-to-anti-fertility' vaccines strands see users as victims of a state-led medical establishment enabled power, which is inscribed in the technology. The more moderate strand of activism argue that women's interests and needs differ from one setting to another, and that they are best met by making available to women a range of contraceptive options which allow for a free and informed choice. PMID- 16039767 TI - Collective efficacy and obesity: the potential influence of social factors on health. AB - Social determinants have been identified as a fundamental cause of health and disease in most industrialized countries. However, much less is known about which characteristics of communities may lead to disparities in health outcomes. Collective efficacy--the willingness of community members to look out for each other and intervene when trouble arises--is a social factor shown to be associated with outcomes related to obesity, including premature mortality and cardiovascular disease. The objective of this study is to determine whether neighborhood collective efficacy is associated with individual measures of body mass index (BMI) in adolescents. We use a multi-level, cross-sectional survey in Los Angeles County, involving 807 adolescents in 684 households in 65 neighborhoods in addition to a sample of 3000 adult respondents. The main outcomes measures are BMI, at risk of overweight, and overweight status. Using a two-level model, we find significant relationships between collective efficacy and all three outcomes, net of levels of neighborhood disadvantage. The associations between BMI and collective efficacy could potentially be explained by several factors, including a metabolic pathway, neighborhood differences in the physical and social environments, or a combination of these two. If group level collective efficacy is indeed important in the regulation of individual level net energy balance, it suggests that future interventions to control weight by addressing the social environment at the community level may be promising. PMID- 16039768 TI - Nutrition-related health promotion through an after-school project: the responses of children and their families. AB - UK health policy is concerned with emphasising nutritional status as a factor in the relationship of social inequalities to health outcomes. This paper examines pupil and parent responses to an after-school 'Food Club' designed to promote food preparation skills and healthier food choices amongst 12-13 year olds in low income areas in North East England. The rationale for the intervention was a series of distinct but connected premises: food preparation skills are essential to inexpensive healthier eating; practical cooking skills are given limited emphasis in the secondary school curriculum; children have some choice over what they eat and may serve as a conduit of influence within the family. The qualitative investigation used group discussions and individual interviews with participating pupils and their parents. Most participating pupils enjoyed the practical emphasis upon food preparation, believed their skills developed, and were aware of the underlying message about healthier eating, but only made limited changes to their diet. Interviews with parents showed most to be positive about their child's involvement in such a club, though they varied in their attitude to its underlying message. There was some evidence of children being more involved in cooking at home and making some specific requests about food, but little to suggest they were influencing family food consumption. The findings suggest that an extra-curricular Food Club is an appropriate and feasible approach to developing food preparation skills with pupils in this age group. These findings raise questions about children's opportunities to exercise food preparation skills and make food choices within families, and the extent to which barriers to dietary change may be lowered through educational initiatives directed at pupils. PMID- 16039769 TI - Anti-vaccination movements and their interpretations. AB - Over the last two or three decades, growing numbers of parents in the industrialized world are choosing not to have their children vaccinated. In trying to explain why this is occurring, public health commentators refer to the activities of an anti-vaccination 'movement'. In the light of three decades of research on (new) social movements, what sense does it make to attribute decline in vaccination rates to the actions of an influential anti-vaccination movement? Two sorts of empirical data, drawn largely from UK and The Netherlands, are reviewed. These relate to the claims, actions and discourse of anti-vaccination groups on the one hand, and to the way parents of young children think about vaccines and vaccination on the other. How much theoretical sense it makes to view anti-vaccination groups as (new) social movement organizations (as distinct from pressure groups or self-help organizations) is as yet unclear. In any event there is no simple and unambiguous demarcation criterion. From a public health perspective, however, to focus attention on organized opponents of vaccination is appealing because it unites health professionals behind a banner of reason. At the same time it diverts attention from a potentially disruptive critique of vaccination practices; the critique in fact articulated by many parents. In the light of current theoretical discussion of 'scientific citizenship' this paper argues that identifying anti-vaccination groups with other social movements may ultimately have the opposite effect to that intended. PMID- 16039770 TI - [3 D evaluation and breast plastic surgery: preliminary study]. AB - Due to the development of the invasive technique of digital exam, the three dimensional results are more and more precise, spectacular and helpful. The 3D surfacic model is for a medical approach and non invasive. Early in 2003, we have started a patient evaluation before and after surgery. The most common surgical techniques from the breast reconstruction to the breast augmentation and the lipomodeling have been analyzed. This preliminary study has shown objectively some surgical rules and volumetric results. Those results are the elements of the future development of the 3D surfacic model in plastic surgery. PMID- 16039771 TI - In vivo effects of chronic contamination with depleted uranium on CYP3A and associated nuclear receptors PXR and CAR in the rat. AB - In addition to its natural presence at high concentrations in some areas, uranium has several civilian and military applications that could cause contamination of human populations, mainly through chronic ingestion. Reports describe the accumulation of this radionuclide in some organs (including the bone, kidney, and liver) after acute or chronic contamination and show that it produces chemical or radiological toxicity or both. The literature is essentially devoid of information about uranium-related cellular and molecular effects on metabolic functions such as xenobiotic detoxification. The present study thus evaluated rats chronically exposed to depleted uranium in their drinking water (1mg/(ratday)) for 9 months. Our specific aim was to evaluate the hepatic and extrahepatic mRNA expression of CYP3A1/A2, CYP2B1, and CYP1A1 as well as of the nuclear receptors PXR, CAR, and RXR in these rats. CYP3A1 mRNA expression was significantly higher in the brain (200%), liver (300%), and kidneys (900%) of exposed rats compared with control rats, while CYP3A2 mRNA levels were higher in the lungs (300%) and liver (200%), and CYP2B1 mRNA expression in the kidneys (300%). Expression of CYP1A1 mRNA did not change significantly during this study. PXR mRNA levels increased in the brain (200%), liver (150%), and kidneys (200%). Uranium caused CAR mRNA expression in the lungs to double. Expression of RXR mRNA did not change significantly in the course of this study, nor did the hepatic activity of CYP2C, CYP3A, CYP2A, or CYP2B. Uranium probably affects the expression of drug-metabolizing CYP enzymes through the PXR and CAR nuclear receptors. These results suggest that the stimulating effect of uranium on these enzymes might lead to hepatic or extrahepatic toxicity (or both) during drug treatment and then affect the entire organism. PMID- 16039772 TI - The C-terminal and beta-wing regions of ammodytoxin A, a neurotoxic phospholipase A2 from Vipera ammodytes ammodytes, are critical for binding to factor Xa and for anticoagulant effect. AB - Ammodytoxin A (AtxA) from the venom of Vipera ammodytes ammodytes belongs to group IIA secreted phospholipase A2 (sPLA2), for which the major pathologic activity is presynaptic neurotoxicity. We show here that this toxin also affects hemostasis because it exhibits strong anticoagulant activity. AtxA binds directly to human coagulation factor Xa (FXa) with Kdapp of 32 nM, thus inhibiting the activity of the prothrombinase complex with an IC50 of 20 nM. To map the FXa interaction site on AtxA, various mutants of AtxA produced by site-directed mutagenesis and expressed in Escherichia coli were tested in the study. In surface plasmon resonance (SPR) measurements, with FXa covalently attached to the sensor chip, we show that the FXa-binding site on AtxA includes several basic amino acid residues at the C-terminal and beta-wing regions of the molecule. Applying an in vitro biological test for inhibition of prothrombinase activity, we further demonstrate that the same residues are also very important for the anticoagulant activity of AtxA. We conclude that the anticoagulant site of AtxA is located in the C-terminal and beta-wing regions of this phospholipase A2. Synthetic peptides comprising residues of the deduced anticoagulant site of AtxA provide a basis to synthesize novel anticoagulant drugs. PMID- 16039773 TI - Comments on the paper by M. Barteri, A. Pala, S. Rotella, Structural and kinetic effects of mobile phone microwaves on acetylcholinesterase activity, Biophys. Chem. 113 (2005) 245-253. PMID- 16039774 TI - The 5alpha-reductase type II A49T and V89L high-activity allelic variants are more common in men with prostate cancer compared with the general population. AB - OBJECTIVES: To compare men with prostate disease with those from the general population regarding polymorphisms in the androgen receptor gene and in the 5alpha-reductase II (SRD5A2) gene. MATERIALS AND METHODS: The SRD5A2 polymorphisms A49T, V89L and R227Q, the androgen receptor CAG and GGN repeats and sex hormone status was investigated in men with prostate cancer (CaP) (n=89), benign prostate hyperplasia (n=45) and healthy military conscripts (n=223). RESULTS: The SRD5A2 high-activity allele variants A49T AT and V89L LL were more frequent in CaP-patients compared to general population, p=0.026 and p=0.05, respectively. CaP progression was, however, independent of SRD5A2 variants. In contrary, men with GGN<23 had a higher risk of dying from the disease than their counterparts with longer repeats. CONCLUSIONS: Men with CaP were more often genetically predisposed to a higher enzymatic activity in the turn over from T to DHT compared to the general population. In our population, androgen receptor genotype affected CaP outcome. PMID- 16039775 TI - Does the size of ureteral stent impact urinary symptoms and quality of life? A prospective randomized study. AB - OBJECTIVE: To evaluate the effect of stent diameter on patients' symptoms and quality of life (QoL) by using dedicated questionnaires. METHODS: We prospectively enrolled 34 patients with unilateral ureteral obstruction due to urinary stone undergoing to ureteral stenting (17 pts with 4.8 F and 17 pts with 6 F) before treatment of stone disease. Twenty-one patients with lower urinary symptoms from other causes were used as a control group. Two questionnaires, one on QoL and another on stent specific symptoms, were administered to patients one week after stent positioning and 4 week after removal. RESULTS: There was a significant association between stent state and answers on pain and discomfort on QoL questionnaire. A high percentage of patients reported anxiety and depression associated with the stent. Similar significant association was found between stent state and urinary symptoms and pain. No differences in QoL and urinary symptoms and pain were detected using stents with different size. CONCLUSIONS: Ureteral stents are invariably associated with urinary symptoms and impaired QoL. We did not find any difference between stent with different size, whereas there was a tendency for stent with smaller diameter to dislodge more often. PMID- 16039776 TI - REM sleep deprivation inhibits LTP in vivo in area CA1 of rat hippocampus. AB - Rapid eye movement (REM) sleep deprivation has previously been shown to interfere with normal learning and memory and to inhibit long-term potentiation (LTP) in vitro. Previous studies on REM sleep deprivation and LTP have relied on in vitro analysis in isolated brain slices taken from animals following several days of sleep deprivation. LTP in the hippocampus in situ may differ from LTP in vitro due to modulatory inputs from other brain regions, which are altered after REM sleep deprivation. Here, we examined LTP in unanesthetized, behaving animals on the first and second recovery days following REM sleep deprivation to determine if similar effects are seen in vivo as previously reported in vitro. We found that LTP was significantly impaired in REM sleep-deprived animals on the second recovery day but not the first recovery day. Our results extend previous findings by showing that REM sleep deprivation continues to affect hippocampal function for more than 24h following the end of deprivation. Our results also suggest the presence of a modulatory process not present in vitro. Our findings are not explained by stress during REM sleep deprivation because equivalent circulating corticosterone levels (an index of stress) were found during both REM sleep deprivation and control treatment. PMID- 16039777 TI - Ndrg2 promotes neurite outgrowth of NGF-differentiated PC12 cells. AB - Ndrg2 is a member of the N-myc downstream-regulated genes. Thus far, two different isoforms of rat Ndrg2 protein, Ndrg2S and Ndrg2L, have been identified. Recently, we have identified rat Ndrg2 as a novel target molecule of antidepressants and ECT. The functional role of Ndrg2 in the central nervous system, however, remains unclear. In the present study, we examined the expression of endogenous Ndrg2, cellular localization of transfected Ndrg2 protein, and morphological changes resulting from overexpression of Ndrg2 in NGF differentiated PC12 cells. Neurites began to sprout 1-2 days after exposure to NGF; subsequent neurite growth continued for 5 days. During this time, we evaluated Ndrg2 mRNA expression by real-time quantitative PCR and found that expression significantly increased in a time-dependent manner. Interestingly, V5 conjugated Ndrg2S and Ndrg2L proteins expressed in NGF-differentiated PC12 specifically localized to cell surface membranes and growth cones. Moreover, Ndrg2S and Ndrg2L overexpression promoted neurite elongation in NGF differentiated PC12 cells. In conclusion, our findings offer novel insights into the physiological roles of Ndrg2 in the central nervous system. PMID- 16039778 TI - Goal synchronization of bimanual skills depends on proprioception. AB - The present experiments in Human subjects were designed to test whether proprioceptive feedback plays a role in optimising bimanual synchronization in a goal-oriented familiar task. Goal-synchronization is a typical feature of bimanual everyday skills. The purpose of the study was to disturb proprioceptive signalling by means of vibrating the leading left limb while subjects performed a bimanual task on a drawer manipulandum. Blindfolded subjects reached for and opened the drawer with the left hand while the right hand was reaching for grasping an object as the drawer was fully opened. Discrete events of the task were used to measure movement onset times of pulling and grasping hands and of goal arrival times. A spatial--temporal goal invariance was still present despite asymmetrical limb assignments and subjects were blindfolded. In contrast, when vibration (80 Hz) was applied to the forearm flexors of the leading pulling limb, we found that the interval between the hands at goal reaching was significantly prolonged. This suggests that synchronization is not predetermined entirely by feedforward commands and that proprioceptive feedback is necessary for updating an internal forward model and perhaps also for lower-level corrections in order to ensure covariant limb movements for optimal goal-synchronization. PMID- 16039779 TI - Assessing the agreement between Ostertagia ostertagi ELISA tests performed using the crude adult antigen and the adult and larval stage 4 excretory/secretory antigens. AB - The objective of this study was to determine the agreement between ELISA tests conducted using three O. ostertagia antigens: crude adult worm, larval stage 4 (L4) excretory/secretory (ES) and adult ES. This study was carried out on 289 Holstein cows from five herds in Prince Edward Island and one herd in Nova Scotia. Composite milk samples of these cows were collected (between May and September 2002) from the respective provincial laboratories and sent to the Atlantic Veterinary College where each sample was tested for antibodies to O. Ostertagi using an indirect microtitre ELISA test. Results were expressed as optical density ratio (ODR) values. Each milk sample was tested with three ELISA tests, with each test using a different O. ostertagi antigen. There was a slight rise in ODR values of both adult antigens, between May and August, with higher values obtained using the adult ES antigen. L4 ES ODR values were generally higher than those for both adult antigens during the study period, except for May. There was a more dramatic rise in L4 ES ODR values between May and August. Rises in ODR in May and end of July coincided with periods of mass maturation of L4 to adult worms. The results of the study showed that the concordance correlation coefficient (CCC) between tests performed using both ES and the crude antigens were low (crude adult versus adult ES=0.31, crude adult versus L4 ES=0.30). The highest CCC was observed between tests done using both ES antigens (CCC=0.56). Generally, the study results suggest that the antibody response (detectable by the ELISA) is mainly directed against ES antigens (especially L4) than the crude adult worm antigen. PMID- 16039780 TI - Public health aspects of dirofilariasis in the United States. AB - Coin lesions in the human lung present significant differential diagnostic problems to the physician. There are at least 20 known causes of such lesions, including neoplastic lesions, infectious diseases, and granulomas. The human medical literature contains many misconceptions about the life cycle of Dirofilaria immitis, including the method of entry of the infective-stage larvae and the development of the young adult worm. These misconceptions have obscured the recognition of the clinical presentation of pulmonary dirofilariasis and the potential for D. immitis to lodge in many other areas of the human body besides the lung. Exposure to infective larvae of D. immitis is more common in humans than is currently recognized. Reported cases in humans reflect the prevalence in the canine population in areas of the United States. The veterinary literature provides compelling evidence that D. immitis is a vascular parasite, not an intracardiac one. Its presence in the right ventricle is a post-mortem artifact, because it has never been shown to be there by echocardiography or angiography in a living dog, even though these techniques have demonstrated adult D. immitis in the pulmonary, femoral, and hepatic arteries; posterior vena cava; and right atrium of live dogs. Physicians have taken the name "heartworm" literally, believing that the worm lives in the heart and only after it dies does it embolize to the pulmonary artery. However, the coin lesion is spherical in shape, not pyramidal, as embolic infarcts to the lung in humans are known to be. The coin lesion is an end-stage result of the parasite's death in the vascular bed of the lungs and the stimulation of a pneumonitis followed by granuloma formation. This pneumonitis phase of human pulmonary dirofilariasis is often not recognized by the radiologist because of the way pneumonitis is diagnosed and treated and because the developing nodule is obscured by the lung inflammation. Serologic methods for use in humans are needed for clinical evaluations of patients with pneumonitis living in highly enzootic D. immitis regions. As well, epidemiological surveys are needed to determine the real extent of this zoonotic infection. PMID- 16039781 TI - Changes in the EU legislation on Trichinella inspection--new challenges in the epidemiology. AB - The European Union (EU) countries are searching for new ways to certify meat free of Trichinella; however, with the expansion of the EU, the acceptance of a unilateral method is complicated by the variability of pig and human trichinellosis among EU countries, where significantly higher prevalence rates have been observed in the newly added eastern countries. Several attempts have been made to define Trichinella-free areas, but certification of Trichinella-free pig production farms appears to be the only feasible approach. The increasing prevalence of the non-encapsulating species, Trichinella pseudospiralis, in game, domestic pigs and humans has eliminated the compression technique from the new EU legislation to be enacted in 2006. Also, the observation that several species of Trichinella tolerate freezing in horse meat for up to 4 weeks has forced a change in legislation as well where freezing is no longer an option for certifying horse meat. Because current serological detection methods are not suited for meat inspection, classical direct detection methods and inactivation by freezing remain the methods of choice for pork. It has been proposed, therefore, to automate direct inspection methods as a cost effective alternative to certify pig farms free of Trichinella. PMID- 16039782 TI - Time-course of hormonal induction of mitochondrial glycerophosphate dehydrogenase biogenesis in rat liver. AB - Thyroid hormones are important regulators of mitochondrial metabolism. Due to their complex mechanism of action, the timescale of different responses varies from minutes to days. In this work, we studied selective T3 induction of the inner mitochondrial membrane enzyme-glycerophosphate dehydrogenase (mGPDH) in liver of euthyroid rats. We correlated the kinetics of the T3 level in blood, the mRNA level in liver, the activity and amount of mGPDH in liver mitochondria after a single dose of T3. The T3 level reached maximum after 1 h (80 nmol/l) and subsequently rapidly decreased. mGPDH mRNA increased also relatively fast, reaching a maximum after 12 h and fell to the control level after 72 h. An increase of mGPDH activity could be already found after 6 h and reached a maximum after 24 h in accordance with the increase in mGPDH content (2.4-fold vs. 2.7 fold induction). After 72 h, the mGPDH activity showed a significant 30% decrease. When the rats received three subsequent doses of T3, the increase of mGPDH activity was 2-fold higher than after a single T3 dose. The results demonstrate that mGPDH displays rapid induction as well as decay upon disappearance of a hormonal stimulus, indicating a rather short half-life of this inner mitochondrial membrane enzyme. PMID- 16039783 TI - Marjolin's ulcer on frostbite. PMID- 16039784 TI - Effect of systematic empiric treatment with imipenem on the bacterial ecology in a burns unit. AB - This study took place over 18 months and was divided into three 6 month periods. During the first and third periods, the bacterial ecology of the unit was reviewed, including the observation of bacteria which were isolated and led us to prescribe general antimicrobial therapy, and record the subsequent antibiograms that became available. During the second 6 month period, any patient developing an infection due (or possibly due) to a "Gram negative" strain received imipenem (as beta lactam antimicrobial agent), usually combined with tobramycin. The comparison between bacteria and antibiograms isolated during the first and the third periods did not show any increase in multiple resistant bacteria or imipenem resistant strains, including methicillin resistant Staphylococcus aureus (MRSA). During the third period, in comparison with the first, the number of Pseudomonas aeruginosa and Acinetobacter strains was lower; however, the number of Enterobacter and Klebsiella had increased. There was an increase of overall resistance to ticarcillin, but there was no increase in resistance to the other antimicrobials concerned in the study. Therefore, we concluded that wide use of imipenem did not impair the bacterial ecology of the unit, if used with precautions such as high dose regimen, de-escalation, and both pharmacokinetics and ecology monitoring. PMID- 16039785 TI - Elderly patients discharged home from the emergency department with minor burns. AB - OBJECTIVES: To describe the risk factors, etiology and referral patterns of elderly patients treated for minor burns in an urban emergency department (ED). METHODS: A retrospective chart review was conducted of persons aged 65 years and older who were treated for a minor burn and discharged home from the ED. Medical records were reviewed for 77 burn patients that presented over a 6-year period. RESULTS: Burn patients had significant co-morbid medical illness. The etiology of the burns was scalds (58%), contact (27%) and flame (12%). Sixty-eight percent of the burns were cooking related. Heating pads, curling irons or hot pipes accounted for the majority of contact burns. Three percent of burn patients were referred to a home care agency for a home safety evaluation at the time of discharge from the ED. CONCLUSION: Cooking-related activities accounted for the majority of minor burns in this series. Common consumer items or environmental hazards were responsible for most contact burns. Elderly patients seen in the ED with minor burns were rarely referred to a home care agency. PMID- 16039786 TI - Amygdalar vasopressin mRNA increases in acute cocaine withdrawal: evidence for opioid receptor modulation. AB - In humans, stress is recognized as a major factor contributing to relapse to drug abuse in abstinent individuals; drugs of abuse themselves or withdrawal from such drugs act as stressors. In the animals, evidence suggests that centrally released arginine vasopressin in both amygdala and hypothalamus plays an important role in stress-related anxiogenic behaviors. The stress responsive hypothalamic-pituitary adrenal axis is under tonic inhibition via endogenous opioids, and cocaine withdrawal stimulates hypothalamic-pituitary-adrenal activity. The present studies were undertaken to determine whether: (1) 14-day (chronic) "binge" pattern cocaine administration (45 mg/kg/day) or its withdrawal for 3 h (acute), 1 day (subacute) or 10 days (chronic) alters arginine vasopressin mRNA levels in amygdala or hypothalamus; (2) the opioid receptor antagonist naloxone (1mg/kg) alters arginine vasopressin mRNA or hypothalamic-pituitary-adrenal hormonal responses in acute cocaine withdrawal; and (3) there are associated changes of mu opioid receptor or proopiomelanocortin mRNA levels. In amygdala, arginine vasopressin mRNA levels were unchanged after chronic "binge" cocaine, but were increased during acute cocaine withdrawal. Naloxone completely blocked this increase. Neither chronic cocaine nor its acute withdrawal altered amygdalar mu opioid receptor mRNA levels. The increase in amygdalar arginine vasopressin mRNA levels was still observed after subacute withdrawal, but not after chronic withdrawal. Although hypothalamic-pituitary-adrenal tolerance developed with chronic "binge" cocaine, there were modestly elevated plasma adrenocorticotropin hormone levels during acute withdrawal. While naloxone produced modest adrenocorticotropin hormone elevations in cocaine-naive rats, naloxone failed to elicit an adrenocorticotropin hormone response in cocaine-withdrawn rats. In hypothalamus, neither chronic cocaine nor acute withdrawal altered arginine vasopressin, proopiomelanocortin or mu opioid receptor mRNA levels. These results show that: (1) opioid receptors mediate increased amygdalar arginine vasopressin gene expression during acute cocaine withdrawal, and (2) cocaine withdrawal renders the hypothalamic-pituitary-adrenal axis insensitive to naloxone. Our findings suggest a potential role for amygdalar arginine vasopressin in the aversive consequences of early cocaine withdrawal. PMID- 16039787 TI - The chick embryo appears as a natural model for research in beta-amyloid precursor protein processing. AB - This study reveals that the chick embryo has active the machinery for the production and degradation of the amyloid beta peptide characteristic of Alzheimer's disease. We cloned the principal beta-amyloid precursor protein isoforms in the chick embryo and observed that they are highly homologous to the human sequences and identical at the C-terminal sequence, including the amyloid beta domain. Mammals such as rat or mouse, more commonly used as animal models of human diseases, have a distinct amyloid beta sequence. The distribution of beta amyloid precursor protein isoforms in the chick embryo revealed that, as in humans, their expression is ubiquitous and the prototype beta-amyloid precursor protein-695 predominated in the nervous system. We also found that the chick embryo expresses the genes for the main proteolytic proteases implicated in the production of amyloid beta, including BACE-1, BACE-2, presenilin-1, presenilin-2 and nicastrin, as well as the amyloid beta-degrading enzyme neprilysin, or ADAM 17, a protease implicated in the non-amyloidogenic processing of beta-amyloid precursor protein. We have also found that between amyloid beta40 and amyloid beta42, this latter seems to be the major amyloid beta peptide produced during chick embryogenesis. The chick embryo appears as a suitable natural model to study cell biology and developmental function of beta-amyloid precursor protein and a potential assay system for drugs that regulate beta-amyloid precursor protein processing. PMID- 16039788 TI - Differential effects of infralimbic cortical lesions on temperature and locomotor activity responses to feeding in rats. AB - The time of food availability induces important behavioral and metabolic adaptations. Animals subjected to feeding restricted to a few daytime hours show increased locomotor activity and body temperature in anticipation of mealtime. In addition, animals under ad libitum feeding show a marked postprandial raise in body temperature and in thermogenesis. The areas of the brain commanding these responses to food are partially known. We investigated in the rat the role of the infralimbic area, located in the medial prefrontal cortex, and considered a visceral-autonomic motor area, in the responses to ad libitum or restricted feeding schedule. We performed infralimbic cortex excitotoxic lesions using injections of ibotenic acid, and measured body temperature and locomotor activity by telemetry in rats under ad libitum and restricted feeding conditions. We found that bilateral infralimbic area lesions prevented both the anticipatory and the postprandial increases in core temperature, decreased mean temperature by nearly 0.3 degrees C during both light/dark phases, and increased daily temperature variability. In contrast, the lesion caused a rapid induction of the anticipatory locomotor activity. These results show that behavioral and metabolic responses to the time of food availability are commanded separately and that the infralimbic area is a key structure to adjust the body temperature to an upcoming meal. PMID- 16039789 TI - Ciliary neurotrophic factor-immunoreactivity in olfactory sensory neurons. AB - Ciliary neurotrophic factor (CNTF) has been implicated in processes of neuroprotection, axonal regeneration and synaptogenesis in the lesioned CNS. In the olfactory system, which is characterized by particularly robust neuroplasticity throughout life, the concentration of CNTF is high even under physiological conditions. In the present study, the cellular localization of CNTF immunoreactivity was studied in the rat and mouse olfactory epithelium. In both species, individual olfactory sensory neurons (ONs) displayed intense CNTF immunoreactivity. The number of CNTF-ir ONs varied interindividually in rats and was lower in mice than in rats. In olfactory epithelia of mice expressing beta galactosidase under control of the CNTF promoter, cells of the ON layer were immunoreactive for the reporter protein. CNTF-ir ONs were olfactory marker protein-positive and growth associated protein 43-negative. CNTF-ir ONs lacked apoptotic markers, and the number of specifically labeled ONs was apparently unchanged after light chemical lesioning of the epithelium, indicating that CNTF immunoreactivity was not associated with ON death. Electron microscopy of CNTF-ir ON axons in innervated olfactory bulb glomeruli documented that they formed typical ON axonal synapses with target neurons. Three dimensional reconstructions of bulb pairs showed a striking similarity of the positions of glomeruli innervated by CNTF-ir ON axons in left and right bulbs of individual animals and interindividually. The number of innervated glomeruli differed interindividually in rats and was lower in mice than in rats. The results show that in rodents CNTF immunoreactivity occurs in a subset of mature, functionally competent ONs. The localization of target glomeruli suggests that CNTF-immunoreactivity may be associated with the expression and/or activation of specific olfactory receptor proteins. PMID- 16039790 TI - Deficits in cortico-striatal synaptic plasticity and behavioral habituation in rats with portacaval anastomosis. AB - Hepatic encephalopathy is characterized by disturbances of motor and cognitive functions involving the basal ganglia. So far no standards for assessment of neuropsychiatric abnormalities (disorders of sleep, mood, anxiety and personality) in subclinical hepatic encephalopathy have been defined. Using an animal model of mild (subclinical) hepatic encephalopathy we investigated now striatum-related behaviors and cortico-striatal synaptic plasticity in rats 2 months after introduction of a portacaval shunt and sham-operated matched controls. In a novel open field portacaval shunt rats displayed less locomotor activity; unlike controls they also showed no habituation to the field and no recall of the field environment after 24 h, indicative of cognitive deficit. The elevated-plus maze test indicated no differences in fear/anxiety in the portacaval shunt animals. Tetanic stimulation of cortical afferents in magnesium free solution evoked an N-methyl-D-aspartate-dependent long-term potentiation in sham-operated animals. In portacaval shunt animals long-term potentiation was significantly impaired. Histamine, a potent modulator of cortico-striatal transmission, induced a larger long-term depression of field potentials in control compared with portacaval shunt rats. In conclusion, a combination of electrophysiological and behavioral approaches has revealed functional changes in cortico-striatal transmission. These data are relevant for understanding the mechanisms of motor and cognitive dysfunctions in hepatic encephalopathy patients and for the development of precise psychometric tests, evaluating cognitive deficits in subclinical hepatic encephalopathy. PMID- 16039791 TI - Signaling pathways of interleukin-1 actions in the brain: anatomical distribution of phospho-ERK1/2 in the brain of rat treated systemically with interleukin 1beta. AB - Interleukin-1beta is released at the periphery during infection and acts on the nervous system to induce fever, neuroendocrine activation, and behavioral changes. These effects are mediated by brain type I IL-1 receptors. In vitro studies have shown the ability of interleukin-1beta to activate mitogen-activated protein kinase signaling pathways including p38, c-Jun N-terminal kinase and extracellular signal-regulated protein kinase 1 and 2 (ERK1/2). In contrast to other mitogen-activated protein kinases, little is known about ERK1/2 activation in the rat brain in response to interleukin-1beta. The aim of the present study was therefore to investigate spatial and temporal activation of ERK1/2 in the rat brain after peripheral administration of interleukin-1beta using immunohistochemistry to detect the phosphorylated form of the kinase. In non stimulated conditions, phosphorylated ERK1/2 immunoreactivity was observed in neurons throughout the brain. Administration of interleukin-1beta (60 microg/kg, i.p.) induced the phosphorylation of ERK1/2 in areas at the interface between brain and blood or cerebrospinal fluid: meninges, circumventricular organs, endothelial like cells of the blood vessels, and in brain nuclei involved in behavioral depression, fever and neuroendocrine activation: paraventricular nucleus of the hypothalamus, supraoptic nucleus, central amygdala and arcuate nucleus. Double labeling of phosphorylated ERK1/2 and cell markers revealed the expression of phosphorylated ERK1/2 in neurons, astrocytes and microglia. Since phosphorylated ERK1/2 was found in structures in which type I IL-1 receptor has already been identified as well as in structures lacking this receptor, activation of ERK1/2 is likely to occur in response to both direct and indirect action of interleukin-1beta on its target cells. PMID- 16039792 TI - Influence of feeding status on neuronal activity in the hypothalamus during lipopolysaccharide-induced anorexia in rats. AB - Fasting attenuates disease-associated anorexia, but the mechanisms underlying this effect are not well understood. In the present study, we investigated the extent to which a 48 h fast alters hypothalamic neuronal activity in response to the anorectic effects of lipopolysaccharide in rats. Male rats were fed ad libitum or fasted, and were injected with i.p. saline or lipopolysaccharide (250 microg/kg). Immunohistochemistry for Fos protein was used to visualize neuronal activity in response to lipopolysaccharide within selected hypothalamic feeding regulatory nuclei. Additionally, food intake, body weight, plasma interleukin-1 and leptin levels, and the expression of mRNA for appetite-related neuropeptides (neuropeptide Y, proopiomelanocortin and cocaine-amphetamine-regulated transcript) were measured in a time-related manner. Our data show that the pattern of lipopolysaccharide-induced Fos expression was similar in most hypothalamic nuclei whatever the feeding status. However, we observed that fasting significantly reduced lipopolysaccharide-induced Fos expression in the paraventricular nucleus, in association with an attenuated lipopolysaccharide induced anorexia and body weight loss. Moreover, lipopolysaccharide reduced fasting-induced Fos expression in the perifornical area of the lateral hypothalamus. Lipopolysaccharide-induced circulating levels of interleukin-1 were similar across feeding status. Finally, fasting, but not lipopolysaccharide, affected circulating level of leptin and appetite-related neuropeptides expression in the arcuate nucleus. Together, our data show that fasting modulates lipopolysaccharide-induced anorexia and body weight loss in association with neural changes in specific hypothalamic nuclei. PMID- 16039794 TI - Urocortin 1-containing neurons in the human Edinger-Westphal nucleus. AB - The topographical location of neurons containing urocortin 1, a peptide related to corticotropin-releasing factor was investigated in human postmortem brain by immunohistochemistry, and compared with the location of neurons containing choline acetyltransferase, a marker for cholinergic cells. A three-dimensional computer reconstruction of the urocortin 1 and choline acetyltransferase-positive population of neurons within the oculomotor area was made. It was shown that the urocortin 1-positive neurons are located within the area identified as the Edinger-Westphal nucleus according to the human brain stem atlas, and that the neurons identified as Edinger-Westphal nucleus in the atlas are not choline acetyltransferase-positive. This finding agrees with recent animal studies showing that urocortin 1-positive neurons are not identical with the parasympathetic cholinergic neurons projecting to the ciliary ganglion. They indicate that the neurons identified as Edinger-Westphal nucleus in the human brain stem atlas belong to the non-preganglionic Edinger-Westphal nucleus, whereas the location of preganglionic Edinger-Westphal nucleus remains unidentified. PMID- 16039793 TI - Developmental emergence of hippocampal fast-field "ripple" oscillations in the behaving rat pups. AB - Sharp wave and associated fast oscillatory ripples (140-200 Hz) in the cornu ammonis 1 region are the most synchronous hippocampal patterns in the adult rat. Spike sequences associated with sharp waves are believed to play a critical role in transferring transient memories from the hippocampus to the neocortex and the emergence of superfast ripples is pathognostic in temporal lobe epilepsy. Sharp waves in cornu ammonis 1 stratum radiatum are induced by a strong depolarization by the cornu ammonis 3 Schaffer collaterals, due to the synchronous discharge of cornu ammonis 3 pyramidal cells. Although during the first postnatal week, sharp wave events are associated with hippocampal unit bursts in the pyramidal layer, ripple oscillations are absent. We investigated the emergence of fast-field oscillations in rat pups ranging from postnatal day 12-20 by recording with wire tetrodes in freely behaving pups and with 16-site linear silicon probes in head fixed animals. Cornu ammonis 1 pyramidal cell layer was determined by the presence of multiple unit activity and a reversal of the field potential in the deeper electrode sites. On-line verification of the recording sites was determined via an evoked response to commissural stimulation, showing a clear reversal in the field potential. Sharp wave-associated fast-field oscillations did not begin to emerge until the end of the second postnatal week and showed a gradual increase until day 18. Once ripples emerged, the intra-ripple frequency assumed adult values. The developmental time course of the ripple parallels the switch in the GABA(A) receptor-mediated signaling from excitation to inhibition. The time course may also reflect hitherto unidentified emergence of neuronal gap junctions. PMID- 16039795 TI - Chemical mediators enhance the excitability of unmyelinated sensory axons in normal and injured peripheral nerve of the rat. AB - Ectopic excitation of nociceptive axons by chemical mediators may contribute to symptoms in neuropathic pain. In this study, we have measured the excitability of unmyelinated rat C-fiber axons in isolated segments of sural nerves under different experimental conditions. (1) We demonstrate in normal rats that several mediators including ATP, serotonin (5-HT), 1-(3-chlorophenyl)biguanide (5-HT3 receptor agonist), norepinephrine, acetylcholine and capsaicin alter electrophysiological parameters of C-fibers which indicate an increase of axonal excitability. Other mediators such as histamine, glutamate, prostaglandin E(2) and the cytokines tumor necrosis factor alpha, interleukin-1beta and interleukin 6 did not produce such effects. (2) The effects of several mediators were tested after peripheral nerve injury (partial ligation or spared nerve injury). Sural nerves from such animals did not show significant changes when compared with controls. (3) We tested whether the effects of chemical mediators on axonal excitability are due to actions on the sensory C-fiber afferents or the postganglionic sympathetic efferents. In order to distinguish these effects, we performed surgical sympathectomy of the lumbar sympathetic chain, including the L3, L4 and L5 ganglia. Sympathectomy did not markedly influence the effects of mediators on axonal excitability (except that the norepinephrine effect was significantly diminished). In conclusion, our data suggest a constitutive rather than inducible expression of axonal receptors for some chemical mediators on the axonal membrane of unmyelinated fibers. Most of the changes in axonal excitability take place in sensory C-fiber afferents rather than in postganglionic sympathetic efferents. Thus, it is possible that certain immune and glial cell mediators released in or around the nerve following injury or inflammation influence the excitability of intact nociceptive fibers. This mechanism could contribute to ectopic excitation of axons in neuropathic pain. PMID- 16039796 TI - Vascular endothelial growth factor and its receptor Flk-1 are expressed in the hippocampus following entorhinal deafferentation. AB - Vascular endothelial growth factor (VEGF) has been thought of as a mitogen that promotes proliferation of endothelial cells and as a neurotrophic factor that stimulates neurogenesis and axonal growth in both peripheral and central nervous systems. To investigate the potential involvement of VEGF in the lesion-induced reorganization in the brain, the expression changes of VEGF and its receptor Flk 1 were analyzed in the mouse hippocampus after transections of the entorhinal afferents. In situ hybridization and immunohistochemistry showed the time dependent expression upregulation of VEGF mRNA and protein in the entorhinally denervated hippocampal stratum lacunosum-moleculare and dentate outer molecular layer, which initiated by 3 days postlesion, reached its maximum at 7-15 days postlesion, still persisted by 30 days postlesion for protein, and recovered to the normal levels at 30 days postlesion for mRNA and at 60 days postlesion for protein. Double labeling of VEGF and glial fibrillary acidic protein revealed that VEGF-expressing cells in the denervated areas were reactive astrocytes. Semi quantitative RT-PCR analysis showed that VEGF receptor Flk-1 mRNA was also time dependently upregulated in the deafferented hippocampus with its maximal elevation at 7-15 days postlesion while the Flt-1 mRNA levels remained unchanged at any time point we examined. Immunohistochemistry analysis also displayed the upregulation of Flk-1 protein in the denervated stratum lacunosum-moleculare and outer molecular layer with a time course similar to that of VEGF mRNA upregulation. Flk-1 receptors were found to be expressed not only by reactive astrocytes but also by neurites, which most likely belong to sprouting axons by 7 days postlesion and regrowing dendrites by 15-30 days postlesion. From these data we suggest that the spatiotemporal upregulation of VEGF and Flk-1 in the hippocampus is induced by entorhinal deafferentation and that VEGF may be involved in the structural reorganization in the deafferented hippocampus via directly or indirectly promoting neurite growth. PMID- 16039797 TI - RFamide-related peptides signal through the neuropeptide FF receptor and regulate pain-related responses in the rat. AB - The mammalian RFamide-related peptide RFRP1 was found to signal through the neuropeptide FF 2 receptor expressed in Xenopus oocytes. The peptide induced a dose-dependent outward current, which was dependent on the simultaneous expression of GIRK1 and GIRK4 potassium channels. In neuropathic rats, RFRP1 administered intrathecally induced tactile antiallodynia and thermal antinociception, whereas in the solitary tract nucleus it produced only mechanical antihyperalgesia. Expression of the RFamide-related peptide mRNA in the rat CNS was distinctly different from that of neuropeptide FF. Most notably, the gene was not expressed in the hindbrain or spinal cord at detectable levels. However, there was a prominent group of RFamide-related peptide mRNA-expressing neurons in the central hypothalamus, in the area in and between the dorsomedial and ventromedial nuclei. The results suggest that RFamide-related peptides are potentially involved in pain regulation through a hypothalamo-medullary projection system, and possibly via action on neuropeptide FF 2 receptors. In neuropathic animals, the pain suppressive effect of RFamide-related peptide varies depending on the submodality of noxious test stimulation and the site of RFamide-related peptide administration. PMID- 16039798 TI - Postnatal development of ectopic sensory fibers containing endomorphin-2 in the white matter of the spinal cord of a transgenic mouse expressing nerve growth factor in oligodendrocytes. AB - Transgenic mice ectopically expressing nerve growth factor in oligodendrocytes have high levels of nerve growth factor immunoreactivity in the white matter of the spinal cord from birth until 2 months of age. The nerve growth factor over expression leads to the appearance of ectopic substance P containing sensory fibers in the white matter of the spinal cord that persist throughout the life of the animal. These transgenic mice have been found to display hypersensitivity to a thermal stimulus following a sensitizing pinch stimulus known to release endogenous substance P. Surprisingly, this hypersensitivity is completely reversed following the administration of morphine, to the extent that transgenic mice become less sensitive to pain than the wild type mice given morphine. Endomorphin-2, an endogenous opioid peptide, has been found co-localized with substance P in primary sensory fibers in the spinal cord. In this study, we show that the ectopic fibers also express endomorphin-2, and describe the postnatal development of such expression, as detected by immunocytochemistry. We confirmed that endomorphin-2 expression starts later in the postnatal period than substance P. Surprisingly, transgenic animals had delayed appearance of endomorphin-2 in the superficial dorsal horn, compared with wild type, and expressed particularly high levels of endomorphin-2 immunoreactivity in the ectopic fibers from postnatal days 10-30, coinciding with the peak of nerve growth factor expression in oligodendrocytes. Endomorphin-2 immunoreactivity was still readily detected in ectopic fibers of 120-day-old animals. Furthermore, we detected immunoreactivity for the mu-opioid receptor in the ectopic fibers, where it was co-localized with endomorphin-2 immunoreactivity. In the superficial dorsal horn, there were no apparent differences in the distribution and intensity of mu-opioid receptor immunoreactivity between wild type and transgenic animals. Taken together, these data could provide an explanation for the enhanced effect of opioid analgesics in transgenic mice, when compared with control mice, as well as provide the basis for studies of the postnatal development of the hyperalgesia and allodynia demonstrated by these animals. PMID- 16039799 TI - Heart rate dynamics and behavioral responses during acute emotional challenge in corticotropin-releasing factor receptor 1-deficient and corticotropin-releasing factor-overexpressing mice. AB - The role of corticotropin-releasing factor in autonomic regulation of heart rate, heart rate variability and behavior responses was investigated in two genetic mouse models: corticotropin-releasing factor receptor 1-deficient mice, and corticotropin-releasing factor-transgenic mice overexpressing corticotropin releasing factor. Heart rate was recorded by radio-telemetry during novelty exposure and auditory fear conditioning. Locomotor activity and freezing served as behavioral indices. Locomotor activity and heart rate were invariably increased in response to novelty exposure in both corticotropin-releasing factor receptor 1-deficient mice and littermate wild-type controls. The heart rate responses during retention of conditioned auditory fear and the exponential relationship between heart rate and heart rate variability were unaffected by genotype. Moreover, conditioned fear responses inferred from multiple behavioral measures including freezing did not differ between corticotropin-releasing factor receptor 1-deficient and corticotropin-releasing factor receptor 1 wild-type control mice. Corticotropin-releasing factor-transgenic mice exhibited markedly reduced locomotor activity during novelty exposure when compared with littermate wild-type controls. Baseline and novelty-driven heart rate was slightly elevated in corticotropin-releasing factor-transgenic mice, whereas the novelty-induced increase of heart rate was not different between genotypes. In contrast, corticotropin-releasing factor-transgenic mice did not display a heart rate response indicative of conditioned auditory fear. It is concluded that corticotropin-releasing factor receptor 1-deficiency does not affect heart rate adjustment and behavioral responses to acute fearful stimuli. The resiliency of behavioral and cardiovascular patterns elevation argues against the involvement of corticotropin-releasing factor receptor 1 in acute emotional regulation on these two functional levels despite an absent corticosterone elevation in corticotropin-releasing factor receptor 1-deficient mice. It is hypothesized that the lack of a conditioned heart rate response in corticotropin-releasing factor transgenic mice is attributable to an impairment of cognitive function. The results are compared with those of corticotropin-releasing factor receptor 2 deficient mice, and the role of the corticotropin-releasing factor system in cardiovascular regulation is discussed. PMID- 16039800 TI - Altered glutamate and GABA release within thalamocortical circuitry in metabotropic glutamate receptor 4 knockout mice. AB - The metabotropic glutamate receptor 4 is highly expressed presynaptically on thalamocortical neurons that are involved in the pathogenesis of generalized absence seizures. Mutant mice devoid of metabotropic glutamate receptor 4 are completely resistant to absence seizures induced by low doses of GABA type A receptor antagonists. The purpose of this study was to test the hypothesis that there is altered glutamate and GABA release within thalamocortical circuitry in mice devoid of metabotropic glutamate receptor 4. Extracellular GABA and glutamate release were determined in ventrobasal thalamus, the nucleus reticularis thalami and laminae I-III, and IV-VI of cerebral cortex (laminae I III of cerebral cortex, and laminae IV-VI of cerebral cortex) using in vivo microdialysis techniques on awake, free moving mice. A significant increase of both basal and K(+)-evoked glutamate release was detected in the ventrobasal thalamus, the nucleus reticularis thalami and laminae IV-VI of cerebral cortex of mice devoid of metabotropic glutamate receptor 4 mice. There also was a significant increase in both basal and K(+)-evoked GABA release in the mice devoid of metabotropic glutamate receptor 4, but a significant decrease of GABA release in laminae IV-VI of cerebral cortex. However, there was no alteration of either GABA or glutamate release in laminae I-III of cerebral cortex, cortical laminae that are not involved in absence seizures. These data indicate that deletion of the metabotropic glutamate receptor 4 gene results in a selective perturbation of glutamate and GABA release within the thalamocortical circuitry involved in the pathogenesis of absence seizures. PMID- 16039801 TI - Co-translational protein aggregation after transient cerebral ischemia. AB - Transient cerebral ischemia leads to irreversible translational inhibition which has been considered as a hallmark of delayed neuronal death after ischemia. This study utilized a rat transient cerebral ischemia model to investigate whether irreversible translational inhibition is due to abnormal aggregation of translational complex, i.e. the ribosomes and their associated nascent polypeptides, initiation factors, translational chaperones and degradation enzymes after ischemia. Translational complex aggregation was studied by electron microscopy, as well as by biochemical analyses. A duration of 15 or 20 min of cerebral ischemia induced severe translational complex aggregation starting from 30 min of reperfusion and lasting until the onset of delayed neuronal death at 48 h of reperfusion. Under electron microscopy, most rosette-shaped polyribosomes were relatively evenly distributed in the cytoplasm of sham-operated control neurons. After ischemia, most ribosomes were clumped into large abnormal aggregates in neurons destined to die. Translational complex components consisting of small ribosomal subunit protein 6, large subunit protein 28, eukaryotic initiation factor-3eta, co-translational chaperone heat shock cognate protein 70 and co-chaperone HSP40-Hdj1, as well as co-translational ubiquitin ligase c-terminus of hsp70-interacting protein were all irreversibly clumped into large abnormal protein aggregates after ischemia. Translational components were also highly ubiquitinated. To our knowledge, irreversible aggregation of translational components has not been reported after brain ischemia. This study clearly indicates that ischemia damages co-translational chaperone and degradation machinery, resulting in irreversible destruction of protein synthesis machinery by protein aggregation after ischemia. PMID- 16039802 TI - The endogenous somnogen adenosine excites a subset of sleep-promoting neurons via A2A receptors in the ventrolateral preoptic nucleus. AB - Recent research has shown that neurons in the ventrolateral preoptic nucleus are crucial for sleep by inhibiting wake-promoting systems, but the process that triggers their activation at sleep onset remains to be established. Since evidence indicates that sleep induced by adenosine, an endogenous sleep-promoting substance, requires activation of brain A(2A) receptors, we examined the hypothesis that adenosine could activate ventrolateral preoptic nucleus sleep neurons via A(2A) adenosine receptors in rat brain slices. Following on from our initial in vitro identification of these neurons as uniformly inhibited by noradrenaline and acetylcholine arousal transmitters, we established that the ventrolateral preoptic nucleus comprises two intermingled subtypes of sleep neurons, differing in their firing responses to serotonin, inducing either an inhibition (Type-1 cells) or an excitation (Type-2 cells). Since both cell types contained galanin and expressed glutamic acid decarboxylase-65/67 mRNAs, they potentially correspond to the sleep promoting neurons inhibiting arousal systems. Our pharmacological investigations using A(1) and A(2A) adenosine receptors agonists and antagonists further revealed that only Type-2 neurons were excited by adenosine via a postsynaptic activation of A(2A) adenosine receptors. Hence, the present study is the first demonstration of a direct activation of the sleep neurons by adenosine. Our results further support the cellular and functional heterogeneity of the sleep neurons, which could enable their differential contribution to the regulation of sleep. Adenosine and serotonin progressively accumulate during arousal. We propose that Type-2 neurons, which respond to these homeostatic signals by increasing their firing are involved in sleep induction. In contrast, Type-1 neurons would likely play a role in the consolidation of sleep. PMID- 16039803 TI - Prevalence and trends of alcohol use and misuse among adolescents and young adults in the Netherlands from 1993 to 2000. AB - This study presents an overview of Dutch studies on prevalence of alcohol use and adds findings from our own study on prevalence of drinking among adolescents and young adults aged 12-30 years in the Netherlands. Data were collected as part of a longitudinal study by the Netherlands Twin Register in 1993 (n=3885), 1995 (n=4814), 1997 (n=3772) and 2000 (n=4090). Measures included lifetime alcohol use, frequency of drinking, quantity of drinking, lifetime drunkenness, frequency of drunkenness and problem drinking. The main findings are: (a) alcohol use increased with age until the age of 25, after which it decreased; (b) males exceeded females on all aspects of alcohol use, with exception of the youngest age group and lifetime alcohol use; (c) time trends indicated an increase in frequency and quantity of drinking among 12-15-year-old adolescents during the 1990s; and moreover, (d) 21-25-year-old females drank more frequently, consumed more drinks a week, had more experience with lifetime drunkenness and were drunk more often in 2000 than in 1993. Among 21-25-year-old males, an increase of drunkenness and problem drinking was displayed during the 1990s. PMID- 16039804 TI - Rapid identification of potentially probiotic Bifidobacterium species by multiplex PCR using species-specific primers based on the region extending from 16S rRNA through 23S rRNA. AB - This study aimed at developing a novel multiplex polymerase chain reaction (PCR) primer set for identification of the potentially probiotic Bifidobacterium species B. adolescentis, B. animalis subsp. animalis (B. animalis), B. bifidum, B. breve, B. longum biovar infantis (B. infantis), B. animalis subsp. lactis B. lactis, B. longum biovar longum (B. longum) and B. pseudolongum. The primer set comprised specific and conserved primers and was derived from the integrated sequences of 16S and 23S rRNA genes and the rRNA intergenic spacer region (ISR) of each species. It could detect and identify type strains and isolates from pharmaceuticals or dairy products corresponding to the eight Bifidobacterium species with high specificity. It was also useful for screening of the related strains from natural sources such as the gastro-intestinal tract and feces. We suggest that the assay system from this study is an efficient tool for simple, rapid and reliable identification of Bifidobacterium species for which probiotic strains are known. PMID- 16039805 TI - The mitochondrial genome of the Japanese freshwater crab, Geothelphusa dehaani (Crustacea: Brachyura): evidence for its evolution via gene duplication. AB - The translocation of tRNA genes has occurred frequently among multiple independent arthropod lineages. However, the mechanisms of such gene rearrangement are still unclear. In this study we sequenced the nearly complete mitochondrial genome of the Japanese freshwater crab, Geothelphusa dehaani (Decapoda, Brachyura, Potamidae). Its size is relatively large (app. 24 kb) for higher animal mtDNA and is due to the presence of many intergenic noncoding sequences (IGNs), one of which is a large repeat region (>7.0 kb). Excluding the repeat region and the putative control region (514 bp), the total size of IGNs (1953 bp) is still the largest reported in arthropods. In comparison to the proposed ancestral arrangement for the insect-crustacean clade, G. dehaani displays changes in the positional order for three tRNA genes (trnQ, trnL(uur) and trnH). At the putative former and current locations for trnQ and trnH, there are IGNs with high similarities to corresponding sequences in the ancestral arrangement. This suggests the occurrence of partial duplication, followed by a loss of function for one copy of these two duplicated genes in G. dehaani. Furthermore, three trnLs (one trnL(uur) and two trnL(cun)s) are found in G. dehaani; their sequences are highly similar to each other (70.8-82.5%) and similar to the trnL(cun) of other arthropods. This suggests that duplication of trnL(cun) occurred at least twice, and the current trnL(uur) of G. dehaani arose by a subsequent anticodon alternation. These observations provide direct evidence for the "duplication-random loss" model for mtDNA gene rearrangement. Comparisons with other available Brachyuran data(http://www.ncbi.nlm.nih.gov/genomes/ORGANELLES/6657.html) suggest that two of the three tRNA translocations are unique in G. dehaani. For G. dehaani, where duplications have occurred but deletion of extra sequences is incomplete, the IGNs found at the ancestral or trans-locations are likely to be footprints of relatively recent mitochondrial genome evolution. PMID- 16039806 TI - Solenopsis invicta transferrin: cDNA cloning, gene architecture, and up regulation in response to Beauveria bassiana infection. AB - Transferrin genes from several insects have been shown to be induced in response to bacterial or fungal infection. We were interested to know whether transferrin genes in the red imported fire ant, Solenopsis invicta, are similarly induced by microbial challenge. Hence, the cDNA and structure of a gene exhibiting significant homology to insect transferrins were elucidated for S. invicta. The cDNA was comprised of 2417 nucleotides, excluding the poly(A) tail, with a large open reading frame of 2106 nucleotides. The predicted translation product of the S. invicta tranferrin (SiTf) gene was a 702 amino acid polypeptide with an estimated molecular mass of 77.3 kDa and a pI value of 5.66, characteristics consistent with transferrin proteins. Comparative analysis of genomic and cDNA sequences revealed that the SiTf gene was comprised of 8 exons. Quantitative real time PCR was used to examine the expression of SiTf. Expression of SiTf was induced in worker ants exposed to Beauveria bassiana conidia. Autoclave-killed conidia did not elicit a SiTf induction response from worker ants. Genes, like SiTf, responding to microbe attack or infection may provide a unique approach to assist in the discovery of microbial control organisms for the target insect pest. PMID- 16039807 TI - The roles of positive and negative selection in the molecular evolution of insect endosymbionts. AB - The evolutionary rate acceleration observed in most endosymbiotic bacteria may be explained by higher mutation rates, changes in selective pressure, and increased fixation of deleterious mutations by genetic drift. Here, we explore the forces influencing molecular evolution in Blochmannia, an obligate endosymbiont of Camponotus and related ant genera. Our goals were to compare rates of sequence evolution in Blochmannia with related bacteria, to explore variation in the strength and efficacy of negative (purifying) selection, and to evaluate the effect of positive selection. For six Blochmannia pairs, plus Buchnera and related enterobacteria, estimates of sequence divergence at four genes confirm faster rates of synonymous evolution in the ant mutualist. This conclusion is based on higher dS between Blochmannia lineages despite their more recent divergence. Likewise, generally higher dN in Blochmannia indicates faster rates of nonsynonymous substitution in this group. One exception is the groEL gene, for which lower dN and dN/dS compared to Buchnera indicate exceptionally strong negative selection in Blochmannia. In addition, we explored evidence for positive selection in Blochmannia using both site-and lineage-based maximum likelihood models. These approaches confirmed heterogeneity of dN/dS among codon sites and revealed significant variation in dN/dS across Blochmannia lineages for three genes. Lineage variation affected genes independently, with no evidence of parallel changes in dN/dS across genes along a given branch. Our data also reveal instances of dN/dS greater than one; however, we do not interpret these large dN/dS ratios as evidence for positive selection. In sum, while drift may contribute to an overall rate acceleration at nonsynonymous sites in Blochmannia, variable selective pressures best explain the apparent gene-specific changes in dN/dS across lineages of this ant mutualist. In the course of this study, we reanalyzed variation at Buchnera groEL and found no evidence of positive selection that was previously reported. PMID- 16039808 TI - Study of correlation between lead-induced cytotoxicity and nitric oxide production in PC12 cells. AB - Despite reduction in its exposure, lead remains a major health problem. The primary target of lead toxicity is the central nervous system. The cellular, intracellular and molecular mechanisms of lead neurotoxicity are numerous, such as induction of apoptosis and interfering with Ca2+ dependent enzyme like nitric oxide synthase (NOS). To investigate the cytotoxic effect of lead on rat pheochromocytoma (PC12) cells, as a suitable model for neuroscience study, and possible correlation between lead toxicity and nitric oxide (NO) production, this study was performed. The current results showed that lead could induce cytotoxicity as well as NO production in a dose dependent manner in PC12 cells after 24h. The cytotoxicity was positively correlated with increased NOx (nitrite and nitrate) production in these cells. L-NAME, a NOS inhibitor, treatment (2.5 mM) could reverse this cytotoxicity. It can be concluded that lead-induced cytotoxicity in PC12 cells could partly be mediated by higher NO production. PMID- 16039809 TI - Postmenopausal women with minimal trauma fractures are unapprised of the existence of low bone mass or osteoporosis. AB - OBJECTIVES: To explore barriers to medical care for osteoporosis following a minimal trauma fracture (MTF). MTFs occur from a fall while in a seated, recumbent or standing position; during normal walking, or from a height less than 4 feet. METHODS: Prospective study and focus groups of women with MTF performed in Chicago from June to December of 2003. Study protocol approved by Institutional Review Board (IRB) and all participants signed consent forms. Focus groups were audiotaped and transcribed, transcripts analyzed using Atlas.ti. Survey results analyzed with SPSS 11.5. RESULTS: Twenty nine participants recruited with average age of 74+/-16 years. MTF occurred at the mean age of 61+/ 8 years, with fifteen (15/29) reporting more that one prior MTF. The age distribution was 40-49 years 2 (2/29) participants, 50-59 years 2 (2/29), 60-69 years 6 (6/29), 70-79 years 10 (10/29), 80 and above 9 (9/29). Most participants (21/29, 70%) reported knowledge about osteoporosis obtained from written media. Osteoporotic fractures were rated on a single item Likert Scale (1=not important to 5=most important) as 2.5 (compared with breast cancer 2.3, and myocardial infarction 2.8). Half (16/29) had osteoporosis counseling with their PCP and 9 were receiving medications for bone loss. We observed a positive correlation between osteoporosis counseling and BMD testing (r=0.6, p<0.001), and a trend toward osteoporosis treatment (r=0.372, p=0.09). Half of the participants had reported the occurrence of MTF to their PCP (14/29), however this did not lead to counseling, BMD testing (r=0.07, p=0.78), or treatment (r=-0.14, p=0.53). None of the women believed that low BMD or osteoporosis had contributed to their fracture. Women studied believed that they were "too young" (12/29) to have osteoporosis. CONCLUSION: Women are not receiving adequate information about osteoporosis; they remain unaware of the connection between MTFs and osteoporosis. Post-menopausal women with MTF do not identify osteoporosis as a cause for the fracture, may exhibit ageism and thereby fail to seek adequate medical care. PMID- 16039810 TI - Phase behavior of the microemulsions and the stability of the chloramphenicol in the microemulsion-based ocular drug delivery system. AB - Microemulsion systems composed of Span20/80+Tween20/80+n-butanol+H2O+isopropyl palmitate (IPP)/isopropyl myristate (IPM) were investigated as model systems of drug carriers for eye drops. Effects of chloramphenicol, normal saline, sodium hyaluronate and various oils on the phase behavior were studied. The phase transition was investigated by the electrical conductivity measurements. The electrical conductivity of the microemulsion was affected by the encapsulation of the drug into the system, and the addition of normal saline and sodium hyaluronate. The chloramphenicol is used to treat the diseases such as trachoma and keratitis. However, this drug in the common eye drops hydrolyzes easily. The main product of the hydrolysis is glycols. Here, the chloramphenicol was trapped into the oil-in-water (o/w) microemulsions and its stability was investigated by the high performance liquid chromatography (HPLC) assays in the accelerated experiments of 3 months. Its location in the microemulsion formulations was determined by means of 1H NMR spectroscopy. The results of HPLC revealed that the contents of the glycols in the microemulsion formulations were much lower than that in the commercial eye drops at the end of the accelerated experiments. It implied that the stability of the chloramphenicol in the microemulsion formulations was increased remarkably. The NMR experiments confirmed that the chloramphenicol molecules should be trapped into the hydrophilic shells of the microemulsion drops, which was composed of many oxyethylene groups. The nitro groups of the chloramphenicol molecules were near the alpha2-CH2 of the surfactant molecules and the benzene rings of the chloramphenicol molecules were near the oxyethylene groups of the surfactant molecules. It was this reason that enabled the chloramphenicol molecules in the microemulsions to be screened from the bulk water and its stability to be increased remarkably. PMID- 16039811 TI - The chemical composition and membrane stability activity of some herbs used in local therapy for anemia. AB - The chemical composition and the membrane stabilizing activity of the decoctions of the dry Hibiscus sabdariffa Linn. (Malvaceae) calyx (Sobo in a Nigerian language), Theobroma cacao Linn. (N.O. Sterculiaceae) root (cocoa) and the dry stalk of white and red cultivars of Sorghum bicolor (L.) Moench (Poaceae) were determined. These herbs are often used locally for the management of anemia. Ash content was from 3.5 to 7.2%, tannin 7-46 mg/kg catechin equivalent, ascorbic acid content 250-625 mg/kg and total anthocyanin 57-150 mg/kg cyanidin-3 glucoside equivalent. Phytate was not detected in any sample but alkaloids and saponins were present except in Sobo. The decoctions contained 0.2-2.0% of the herbs' total tannin content while the pH was 3.2-6.7. Iron and zinc content was highest in Sobo while Theobroma cacao root contained the highest calcium and magnesium. Solubility and mineral availability were highest in the decoction of Sobo and lowest but available in Theobroma cacao root decoction. The decoction of Theobroma cacao exhibited a high membrane stabilizing activity while those of Sorghum bicolor and Hibiscus sabdariffa were only average. The study indicated that the herbal decoctions could provide some minerals and also help to stabilize the red blood cells from stress injury. PMID- 16039812 TI - Do not put too much value on conventional medicines. AB - About 80% of the population of the developing countries still use traditional medicines for their health care. The rational use of traditional medicines in primary health care should be based on the Guidelines for the Assessment of Herbal Medicines as developed by the World Health Organization (WHO). The protection of intellectual property on traditional medicines should also be based on the guidelines for the protection of intellectual property on traditional medicines to be developed by the World Intellectual Property Organization (WIPO). The knowledge of traditional medicines hidden and lost should be researched and the loss of natural resources used as traditional medicines should be prevented. As a new emerging field in ethnopharmacology, treatment guidelines and evidence based evaluation guidelines for ethnoergogenics, and protection guidelines of intellectual property for ethnoergogenics should be developed and put into practice according to guidelines of the WHO and WIPO, respectively. PMID- 16039813 TI - Time-course changes in the expression of heme oxygenase-1 in human subcutaneous hemorrhage. AB - To determine the time-course of human subcutaneous hemorrhage, heme oxygenase (HO)-1 expression and macrophage infiltration were observed using an immunohistochemical technique and semiquantitative analysis. The number of immunoreactive cells and the number of all infiltrating cells of each microscopic field were counted, and the ratio of the former to the latter was calculated as the positive cells ratio. An increase in the HO-1-positive cells ratio was observed starting at 3 h after injury, and the maximum ratio was observed 3 days after injury. The pattern of the increase in the macrophage ratio was similar to that of the HO-1-positive cells ratio in the early period after injury. Observation of serial sections revealed that the expression of HO-1 in the cells corresponded to the localization of macrophage. The present results suggest that the determination of HO-1 expression, as derived from macrophages, might be useful for the estimation of the time-course of subcutaneous hemorrhage. PMID- 16039814 TI - Primary osteogenic sarcoma of the ethmoid sinus: a case report. AB - Primary osteogenic sarcoma arising from the ethmoid sinus is an extremely rare condition. We report herein a 50-year-old female presenting with lacrimation. Computed tomography (CT) scan showed tumor calcification, which is a common feature of osteogenic sarcoma. Exploratory ethmoidectomy was performed, and histological examination confirmed the diagnosis of osteogenic sarcoma. Radiation therapy and chemotherapy failed to reduce the tumor size. The patient was finally treated by anterior craniofacial resection and orbital exenteration. Although primary osteogenic sarcoma of the ethmoid sinus is uncommon, the authors emphasize that osteogenic sarcoma should be suspected when tumor calcification was observed on CT scan and that adequate surgical resection with negative surgical margins remains the mainstay of treatment of this lesion. PMID- 16039815 TI - Paradoxical vocal cord motion--a case report. AB - Paradoxical vocal cord motion (PVCM) is an unusual cause of stridor, which is associated with some underlying causes, such as central nervous system lesion, gastroesophageal reflux or psychogenic problem. Once a diagnosis of PVCM is made, acute management with reassurance and sedation instead of aggressive airway intervention is required. Speech therapy, psychotherapy combination with anti reflux medication is considered to be useful in long-term management. We present a 58 year-old male patient who had suffered from several episodes of acute onset of stridor, short of breath and tachypnea since one year ago. He was initially treated as an asthmatic patient with poor response. Aneurysm of ascending aorta by angiography, and mild gastroesophageal reflux with hiatal hernia by panendoscopy were noted. Then, the paradoxical vocal cord motion during inspiration phase was confirmed by flexible fiberoptic nasopharyngoscope after the consultation with an otolaryngologist. The emergency of his air-hunger was relieved quickly after psychological intervention. Now, he is free of stridor attack under anti-reflux therapy and psychotherapy. PMID- 16039816 TI - Clinical efficacy of 5-fluorouracil (5-FU) topical cream for treatment of cholesteatoma. AB - OBJECTIVE: In order to clarify clinical efficacy of commercially available 5 fluorouracil (5-FU) topical cream, a clinical study was conducted. METHODS: Two to three cubic millimetres of 5-FU topical cream (Kyowa, Roche) was applied on 50 cases of various types of cholesteatoma (50 patients) two to five times with the interval of 2 weeks, and its clinical efficacy was evaluated by the criteria we developed. RESULTS: In total, 59% of the cholesteatomas showed good effect, 29% of them showed fair effect, and the effect was poor in the remaining 12%. It was particularly effective in cholesteatomas in the EAC, attic cholesteatomas with an aerated mastoid, and in recurrent-type cholesteatomas. CONCLUSION: 5-FU topical cream appeared effective for the treatment of cholesteatomas. PMID- 16039817 TI - The average values of the nasal anthropometric measurements in 108 young Turkish males. AB - OBJECTIVE: The aim of the study was to describe the average values of the nasal anthropometric measurements in young Turkish male population. METHODS: Study group consisted of 108 volunteer young Turkish males whose age were 18-30 years (mean age 22.31 year). All subjects were students in Yasar Dogu School of Physical Education and Sport of University of Ondokuz Mayis. RESULTS: The means total length and nasal bridge length of the nose were 56.92 and 55.26 mm, respectively. The means of the nasal bridge width and morphologic nose width were 29.74 and 33.63 mm, respectively. The mean width of the anatomic nose was 23.14 mm. The mean width of the nostril floor was 11.00 mm. The means length and width of the ala were 19.93 and 4.81 mm, respectively. The means of length and width of the columella were 9.34 and 5.34 mm, respectively. The mean frontonasal angle was 134.96 degrees and the mean nasolabial angle was 90.32 degrees. CONCLUSION: The nose was the dominant feature in the facial region. Anthropometric measurements related with the nose measured were compared with the available literature. Average values of the nose in this population may be used as a landmark for guidance to plan corrective surgery of the nose. PMID- 16039818 TI - [Escherichia coli meningitis during bacteremia in the Ibrahima-Diop-Mar infectious diseases clinic, Dakar Fann National Hospital Center (Senegal)]. AB - OBJECTIVES: The aim of this study was to describe epidemiological, clinical, and bacteriological aspects of Escherichia coli bacteremia and meningitis in the Ibrahima-Diop-Mar infectious diseases clinic, Dakar Fann National Hospital Center (Senegal). MATERIALS AND METHODS: Data was collected from the bacteriology laboratory and hospitalization files. RESULTS: 57 cases of E. coli bacteremia were reported. Among them, 10 were associated with meningitis. AIDS was diagnosed in 74% of the cases. The global lethality rate was 47% but this rate was higher in cases of associated meningitis (80 vs 37%) and in AIDS patients (50 vs 27%). Ceftriaxone, aztreonam, gentamicin, and ciprofloxacin were active on more than 95% of strains but cotrimoxazole was active on only 49% of the strains. Resistance to cotrimoxazole was higher among E. coli strains isolated from AIDS patients (62 vs 13%). CONCLUSION: The low susceptibility to cotrimoxazole might increase the incidence of E. coli infections among patients with AIDS. It is therefore important to find an alternative to cotrimoxazole chemoprophylaxis. PMID- 16039819 TI - [Surveillance of antimicrobial resistance of bacteria isolated from bloodstream infections: data of the French National Observatory for Epidemiology of Bacterial Resistance to Antibiotics (ONERBA), 1998-2003]. AB - The French national observatory for epidemiology of bacterial resistance to antibiotics (ONERBA) includes numerous networks for the surveillance of bacterial resistance to antibiotics. OBJECTIVE: The aim of this study was to update antimicrobial resistance data of bacterial pathogens isolated from blood cultures. METHOD: Data was collected from several surveillance surveys reviewed by the ONERBA scientific committee during 2003. RESULTS: Gram positive cocci and Gram negative bacilli accounted respectively for 45 and 49% of all bacteria isolated from blood cultures (N=19 882). The frequency of MRSA among S. aureus was 35%. The frequencies of resistance to penicillin, amoxicillin and cefotaxime of S. pneumoniae were 10, 2, and 0.2%, respectively. In 2002, 70% of coagulase negative staphylococci were resistant to methicillin. Only 50 and 60% of E. coli isolates were susceptible to aminopenicillin and a combination amoxicillin clavulanic acid, respectively. Since 2001, the susceptibility of E. coli isolates to ciprofloxacin has been regularly decreasing. Imipenem and ceftazidime were the most active antimicrobial agents against P. aeruginosa with a susceptibility rates below 90%. CONCLUSION: Data provided by the ONERBA should allow to improve the quality of empiric antimicrobial treatments. PMID- 16039820 TI - Pharmaceutical heroin for inhalation: thermal analysis and recovery experiments after volatilisation. AB - Pharmaceutical heroin for inhalation was developed for a clinical trial on co prescription of heroin and methadone to chronic treatment-resistant heroin addicts. Diacetylmorphine base was selected as the active pharmaceutical ingredient for this product with caffeine anhydrate added as an excipient. Differential scanning calorimetry and thermogravimetric analysis showed that addition of caffeine resulted in a lower melting temperature and a higher volatilisation rate for the mixture than for diacetylmorphine base alone. Recovery experiments showed that 40.8+/-5.3% of diacetylmorphine base could be found in smoke condensate after volatilisation of diacetylmorphine-caffeine tablets. All of the caffeine from each tablet was recovered unchanged in the fumes, while 85.6% of the diacetylmorphine from each tablet was recovered, either unchanged in the fumes or as non-volatilised residue. Recovery was found to be reproducible and only small differences were found between the tablet types. The experimental set-up was found to efficiently collect the vapours resulting from heating the powder. Under the tested experimental conditions, no evidence was found that degradation products of diacetylmorphine or caffeine, other than 6 acetylmorphine (5.9%) had volatilised, even though a decomposed residue was present after heating diacetylmorphine-caffeine samples. Diacetylmorphine caffeine was found to be a suitable basis for pharmaceutical heroin to be used by 'chasing the dragon'. PMID- 16039821 TI - (New) Images in obstetric anesthesia: lumbar tattoos. PMID- 16039822 TI - [Residual neuromuscular blockade]. AB - Despite the availability of modern neuromuscular blocking agents with short or intermediate duration of action, incidence of residual neuromuscular blockade remains very high. Evidences have been recently provided that residual curarization must be defined as a train-of-four ratio below 0.9 at the thumb adductor during the recovery period after anaesthesia. Residual curarization may be associated with serious adverse events related to respiratory depression, pharyngeal dysfunction, hypoxemia and prolongation of the length of stay in the recovery room. Appropriate choice of drugs, perioperative monitoring of neuromuscular function and large indications of pharmacological reversal may reduce the incidence of residual curarization and improve the patient's safety in the postoperative setting. PMID- 16039823 TI - Optimization of a model of red blood cells for the study of anti-oxidant drugs, in terms of concentration of oxidant and phosphate buffer. AB - The aggression of erythrocytes by an oxidative stress induces hemolysis. This paper aims to valid a model of erythrocytes in terms of composition of the phosphate buffer solution and of concentration of a well-known oxidant, AAPH. Three compositions of phosphate buffer solution are mixed with three concentrations of oxidant. The influence of these two parameters on hemolysis is independently studied by a variance analysis and a Kruskal-Wallis test when ANOVA is not available. The hemolysis rate increases with time at fixed oxidant concentration, but is not influenced by the composition of the buffer solution. The highest hemolysis rate, 90%, was only measured within 2 h with the highest oxidant concentration. If we retain this concentration of oxidant, the lower concentration of the buffer can by eliminated by a significant less hemolysis and the highest concentration of the buffer can by chosen in regard of the better precision for a similar hemolysis compared to the mean buffer. We hope to study the effect of anti-oxidant agent with such a model of erythrocytes. PMID- 16039824 TI - A genome-wide set of congenic mouse strains derived from DBA/2J on a C57BL/6J background. AB - In the analysis of complex traits, congenic strains are powerful tools because they allow characterization of a single locus in the absence of genetic variation throughout the remainder of the genome. Here, we report the construction and initial characterization of a genome-wide panel of congenic strains derived from the donor strain DBA/2J on the background strain C57BL/6J. For many strains, we have carried out high-density SNP genotyping to precisely map the congenic interval and to identify any contaminating regions. Certain strains exhibit striking variation in litter size and in the ratio of females to males. We illustrate the utility of the set by "Mendelizing" the complex trait of myocardial calcification. These 65 strains cover more than 95% of the autosomal genome and should facilitate the analysis of the many genetic trait differences that have been reported between these parental strains. PMID- 16039825 TI - Major depression and immunity in alcohol-dependent persons. AB - Altered immunity has been associated with both alcoholism and major depression (MD). We investigated the contribution of MD, as well as alcoholism, to in vitro measures of immunity in inner-city alcohol-dependent (SCID-DSM-III-R) persons and community nonabusers, all otherwise in good health. METHODS: Alcohol-dependent persons at an ambulatory alcohol treatment center who did not abuse other substances were studied along with the comparison sample (total n=122). Enumerative and functional immune measures included leukocyte and lymphocyte subsets, mitogen response, natural killer cell activity (NKCA), and granulocytic phagocytosis. RESULTS: Controlling for alcohol dependence, age, gender, racial background, and medical status, MD was associated with decreased phytohemagglutinin (PHA) responses (p<.03), possibly decreased NKCA (p<.08), and increased circulating monocytes (p<.04). Controlling for MD, age, gender, racial background, and medical status, alcohol dependence was associated with decreased circulating B lymphocytes (p<.02), possibly decreased CD56+ (NK) cells (p<.06), and increased monocytes (p<.04). Responses to concanavalin A and pokeweed mitogen, granulocyte functions, and the composition of other leukocyte and lymphocyte subsets showed no evidence of being associated with MD or with alcoholism (p>.1). Secondary analyses exploring factors such as recent alcohol use, cigarette use, and nutrition suggested that these factors accounted for the altered lymphocyte subsets associated with alcoholism and the possibly decreased NKCA with MD. They did not account for the association of MD with increased monocytes and decreased PHA. DISCUSSION: MD-associated immune changes in alcoholics are modest and consistent with those seen in MD without alcoholism. Some MD- and many alcoholism-associated immune effects appear related to factors such as cigarette use and recent alcohol exposure. PMID- 16039826 TI - The use of topical 5% imiquimod during pregnancy: a case series. PMID- 16039827 TI - Fenvalerate modifies T-type Ca2+ channels in mouse spermatogenic cells. AB - Prior to fertilization sperm must undergo the acrosome reaction that is initiated by opening of T-type Ca(2+) channel. Hence, T-type Ca(2+) channels could be a potential target for agents affecting the acrosome reaction. Our previous data have suggested that fenvalerate, a type II pyrethroid, inhibited the acrosome reaction in mice. To elucidate its potential mechanism we investigated fenvalerate's effect on T-type Ca(2+) channels in mouse pachytene spermatocytes using a whole-cell patch clamp technique. Fenvalerate significantly inhibited T type Ca(2+) currents in a concentration-dependent manner. The maximal inhibitory concentration was 1 microM. The inhibitory effect of fenvalerate was slow and irreversible after washout of the drug. The curves of activation and inactivation simultaneously shifted to hyperpolarization, about 14 mV, suggesting the open time of Ca(2+) channel was unchanged. Voltage-dependent gating of Ca(2+) channel indicated a change in permeability to ions that contributed to fenvalerate's inhibition on Ca(2+) current. Taken together with our previous findings, these data suggest that the changes of T-type Ca(2+) currents contribute to the suppression of acrosome reaction and fertilizing ability caused by fenvalerate. PMID- 16039828 TI - Effects of cadmium on memory processes in mice exposed to transient cerebral oligemia. AB - The purpose of the present study was to examine whether the effects of chronic or acute exposure to cadmium on memory processes in mice could be exacerbated by transiently reducing cerebral oxygen supply. Adult mice were subjected to bilateral clamping of the carotid artery (BCCA) for 30 min under anesthesia. Cadmium chloride was administered intraperitoneally after surgery at single doses of: 0.7 mg/kg (low dose), 1.4 mg/kg (high dose), or at a prolonged dose of 0.7 mg/kg for up to 10 days. Long-term memory was evaluated in a step-through passive avoidance task while spatial working memory was evaluated using a Y-maze spontaneous alternation task. BCCA mice injected with the 1.4 mg/kg dose of cadmium exhibited recall deficits in the step-through passive avoidance task. Combined treatment at either dose had no effect on the acquisition of passive avoidance. In the Y-maze task, spontaneous alternation behavior was only impaired in BCCA mice treated with the prolonged cadmium dose. These results indicate that cerebral oligemic hypoxia may alter cadmium neurotoxicity and potentiate the tendency for cadmium-induced memory impairments in the passive avoidance task and spontaneous alternation deficits. PMID- 16039829 TI - Behavioral and neurochemical effects on rat offspring after prenatal exposure to ethanol. AB - The work studied behavioral and neurochemical alterations in 21-day-old pups, from both sexes (26 g on average) born from female Wistar rats administered daily with ethanol (0.5 or 4.0 g/kg, p.o.), for 30 days before mating, and throughout their gestational period. Ethanol administration continued from delivery up to weaning. The open field, elevated plus maze and forced swimming tests were used to evaluate effects of ethanol on locomotion, anxiety and depression, respectively. Binding assays were used to identify dopaminergic (D1- and D2-like) and muscarinic (M1 plus M2) receptors. Results of the plus maze test indicated significant and dose-dependent increases in the number of entrances in the open arms and in the time of permanence in the open arms, in the prenatally ethanol exposed offspring, as compared to controls, indicating an anxiolytic effect. In the open field test, this group presented decreases in spontaneous locomotor activity as well as in the occurrences of rearing and grooming. Offspring also showed dose-dependent increases in their immobility time in the forced swimming test, characterizing despair behavior. Decreases in the hippocampal (D2: 32%; D1: 25%) and striatal (D2: 30%; D1: 52%) dopaminergic binding were detected in ethanol-exposed offspring. On the other hand, significant increases were observed in muscarinic binding in the hippocampus (40%) as well as in the striatum (42%). This study shows evidence that in utero ethanol exposure produces a long-lasting effect on development and pharmacological characteristics of brain systems that may have important implications in behavioral and neurochemical responsiveness occurring in adulthood. PMID- 16039830 TI - Chronic low intake of protein or vitamins increases the intestinal epithelial cell apoptosis in Wistar/NIN rats. AB - OBJECTIVE: Malnutrition decreases antioxidant defense and increases oxidative stress in the intestine. We studied the effects of long-term restriction of food, protein, and vitamins on intestinal epithelial cell (IEC) apoptosis and the underlying mechanisms. METHODS: Weanling, Wistar/NIN male rats were fed ad libitum with a control diet, 75% protein-restricted diet, or 50% vitamin restricted diet for 20 wk. The food-restricted group received 50% of the diet consumed by control rats. IEC apoptosis was monitored by morphometry, Annexin V binding, M30 CytoDeath assay, and DNA fragmentation. Structural and functional integrity of the villus were assessed by the ratio of villus height to crypt depth, and alkaline phosphatase and lys, ala-dipeptidyl aminopeptidase activities, respectively. Oxidative stress parameters, caspase-3 activity, and expression of Bcl-2 and Bax were determined to assess the probable mechanisms of altered apoptosis. RESULTS: Protein and vitamin restrictions but not food restriction significantly increased IEC apoptosis and only vitamin restriction altered structural and functional integrity of villi. Increased levels of protein carbonyls, thiobarbituric acid reactive substances, and caspase-3 activity along with decreased glutathione levels and Bcl-2 expression were observed in IECs of these rats, whereas food restriction did not affect these parameters. CONCLUSIONS: Protein restriction increased only IEC apoptosis, whereas vitamin restriction also affected the structure and function of villi. Modulation of the pathway mediated by mitochondria through increased oxidative stress appears to be the probable mechanism underlying this effect. PMID- 16039831 TI - Evaluation of an iron specific checklist for the assessment of dietary iron intake in pregnant and postpartum women. AB - OBJECTIVE: This study developed and validated an iron checklist for assessing dietary iron intake of pregnant and postpartum women. METHODS: The checklist included 65 food and drink items. Iron intake measured by the checklist was compared with a diet history interview by paired t test and by the Bland-Altman method in 54 pregnant women. We then used the checklist to prospectively assess iron intake in a separate group of women in late pregnancy (n = 179) and at 6 mo postpartum (n = 177). The ability of the checklist to predict iron status was evaluated. RESULTS: There was no difference in reported mean iron intakes between the checklist and the diet history and there were good correlations between iron intake estimated from both methods (r = 0.69, P < 0.001 for food alone, r = 0.99, P < 0.001 for food plus supplements). However, the agreement between the two methods at an individual level was low. Pregnant women with low iron intake (lower than the recommended dietary intake) had lower serum ferritin levels (9.7 versus 14.5 microg/L, P < 0.001) and higher risk of iron deficiency (67.5% versus 34.9%, P < 0.0001) compared with women with adequate iron intake (at least the minimum recommended dietary intake), but these differences disappeared when women taking iron supplements were excluded. There was no association between iron intake and serum ferritin at 6 mo postpartum or between iron intake and hemoglobin levels at the end of pregnancy or at 6 mo postpartum. CONCLUSION: This simple iron checklist is a useful tool in describing iron intake of population samples of pregnant women but has limited ability to predict iron status. PMID- 16039832 TI - Is the national nutrition survey in Japan representative of the entire Japanese population? AB - OBJECTIVE: We examined the representativeness of the sample used in the National Nutrition Survey in Japan (J-NNS). METHODS: The distribution of the J-NNS sample from 1995 to 2000 was compared with that of the national census with respect to gender, age group, place of residence, and number of household members. RESULTS: In the survey sample, as compared with the national census distributions, the proportion of females was 1% to 2% larger, the proportion of individuals who were 20 to 39 y of age was a few percentages smaller, and a slight bias toward rural areas was seen. Among individuals who were 20 to 59 y of age, the proportion of individuals who lived in single-person households was 50% to 66% of that found in the national census. Observed disparities between the sample and the population were minimal except in the case of the proportion of individuals who lived alone. CONCLUSIONS: The sample used in the J-NNS is generally representative of the entire Japanese population with the exception of individuals in single-person households. PMID- 16039833 TI - Neonatal seizures with tonic clonic sequences and poor developmental outcome. AB - Seizures consisting of a tonic followed by a clonic phase have rarely been described in neonates and are not included in the current classifications of neonatal seizures. Our video archive of 105 neonates with seizures or suspected seizures revealed six neonates with such tonic clonic or tonic myoclonic sequences. Two of those neonates had pyridoxine dependent seizures. The other four neonates had drug refractory seizures and demonstrated similarities in electro-clinical pattern, clinical course and outcome. Their seizures started with tonic posturing and after 10-20s tonic posturing was superimposed by focal or multifocal cloni or myocloni. Ictal EEG started with voltage attenuation followed by bilateral or alternating focal epileptic discharges. The interictal EEG was abnormal. One child died, while the other three children became seizure free but had severe motor delay and mental retardation. In one of those three children, a de novo missense mutation was detected in the voltage gated potassium channel gene KCNQ2, indicating a genetic relationship between drug refractory neonatal seizures of unknown etiology with tonic clonic or myoclonic sequences and the well-known syndrome of benign familial neonatal convulsions (BFNC). PMID- 16039834 TI - Association study of the DISC1/TRAX locus with schizophrenia in a Japanese population. AB - Disrupted-in-Schizophrenia-1 (DISC1), identified by cytogenetic approaches in a pedigree with familial psychosis, is considered a candidate susceptibility gene for schizophrenia in some populations. In the pedigree, the TRAX gene, located adjacent to DISC1 on the disrupted chromosome 1, may also contribute to the pathophysiology of the familial schizophrenia. We studied association of the DISC1 and TRAX genes with schizophrenia in 338 Japanese by analyzing 15 single nucleotide polymorphisms (SNPs), including 12 SNPs in DISC1 and three in TRAX, respectively. No significant difference was observed between the patients and controls in allelic frequencies or genotypic distributions of 15 SNPs. A weak trend for the association in genotypic distribution of one SNP in TRAX (major homo/hetero/minor homo: 0.324/0.431/0.245 vs. 0.293/0.526/0.181 for patients vs controls, p = 0.039 in the 2 x 3 comparison) turned out to be insignificant after Bonferroni correction. Haplotype analysis did not support the association between the patients and controls. The present study suggests that the DISC1/TRAX locus may not have a major role in Japanese schizophrenia. PMID- 16039835 TI - The role of low-density lipoprotein receptors in sensitivity to killing by Photofrin-mediated photodynamic therapy in cultured human tumor cell lines. PMID- 16039836 TI - Methotrexate-loaded biodegradable polymeric micelles: preparation, physicochemical properties and in vitro drug release. AB - Polymeric micelles based on amphiphilic diblock copolymers methoxy poly(ethylene glycol)-polylactide with various hydrophobic lengths were designed as carriers of poorly water-soluble anticancer drug methotrexate (MTX). Relationship between physicochemical characteristics of micelles and release behavior was explored. The critical micelle concentration was determined by fluorescence spectroscopy using 9-chloromethyl anthracene as fluorescence probe. Core-shell type polymeric micelles were prepared by free-surfactant dialysis technique. The mean size of micelles loaded with MTX was 50-200 nm with narrow polydispersity. Physicochemical properties of drug-loaded micelles were evaluated. In vitro release behavior of MTX was also investigated. MTX was continuously released from micelles and less than 50% MTX was released in 5 days. Release rate was dependent on chemical structures of micelles and enhanced by decreasing polylactide lengths. PMID- 16039837 TI - [First Congolese case report of Caffey disease]. PMID- 16039838 TI - Health-related quality of life in patients with chronic respiratory failure after long-term mechanical ventilation. AB - STUDY OBJECTIVES: This study was aimed at assessing health-related quality of life (HRQL) in patients with chronic respiratory failure (CRF) and long-term survival following prolonged intensive care mechanical ventilation. DESIGN: Observational cohort study. SETTING: Patients with CRF who had been transferred to our specialized weaning centre due to prolonged mechanical ventilation (>14 days) and weaning failure. PATIENTS AND PARTICIPANTS: Out of 87 long-term survivors (>6 months), 73 patients (mean age: 60.3+/-13.6 years, chronic obstructive pulmonary disease (COPD, 43%), thoraco-restrictive (21%) or neuromuscular disorders (15%), various chronic diseases (22%)) returned the MOS 36-Item Short-Form Health Status Survey (SF-36) and the St. George's respiratory questionnaire (SGRQ). MEASUREMENTS AND RESULTS: The total ventilation time was 38.7+/-45.9 days. The time between discharge from ICU and HRQL assessment was 31.0+/-22.2 months. Physical health was markedly reduced compared to general population norm, but mental health was mildly impaired. HRQL was comparable to patients with stable CRF receiving non-invasive ventilation who did not need prolonged invasive MV. In addition, general HRQL was better in patients with restrictive respiratory disease compared to patients with neuromuscular diseases (P<0.05). Physiological parameters such as blood gases or lung function parameters were not correlated to any HRQL measurements. CONCLUSIONS: In patients with CRF surviving prolonged ventilation on ICU, the presence of CRF itself is the major determinant of HRQL. Here, the underlying cause of CRF is the major factor which determines the degree of HRQL impairment with patients suffering from restrictive ventilatory disorders reporting the best HRQL when compared to patients with COPD or neuromuscular diseases. Despite severe physical handicaps due to CRF mental health is only mildly compromised. PMID- 16039839 TI - Adolescent respiratory symptoms--girls are at risk: the Young-HUNT study, Norway. AB - The objective was to study sex differences in adolescence regarding prevalence of asthma and current wheeze and to explore the association between respiratory symptoms and hereditary, lifestyle and socioeconomic factors. Young-HUNT included data comprehensive questionnaire on health, disease, lifestyle and social factors from 8817 teenagers 13-19 years conducted in 1995/97 (89% response rate). Questionnaire on respiratory symptoms was based on the International Study of Asthma and Allergy in Childhood (ISAAC). In age groups 13-16 and 17-19 years, current wheeze was reported by 29.0% and 33.5% among girls and 20.4% and 22.1% among boys, whilst the corresponding figures for asthma were 8.5% and 12.2% among girls and 7.1% and 7.0% among boys. Both wheeze and asthma were significantly more prevalent and increased with age in girls compared to boys. Heredity was associated with asthma, but the association was strongest between parents and children of the same sex. Environmental smoking was associated with asthma and wheeze in girls only. Girls reported more asthma and wheeze in association with overweight compared to boys. Girls reported more wheeze and asthma than boys and seemed more susceptible to risk factors such as environmental smoking and overweight than boys. Moreover, girls with mothers having asthma were more likely to be diagnosed as asthmatics themselves. PMID- 16039840 TI - Use of medication and quality of life among patients with COPD. AB - STUDY OBJECTIVES: To determine the medications prescribed to patients with chronic obstructive pulmonary disease (COPD) and their relationship to health related quality of life (HRQL). METHODS: Cross-sectional study of 611 consecutive patients with stable, mild-to-severe COPD who attended at the respiratory service of a single hospital during a 1-year period. HRQL was evaluated using the St. George Respiratory Questionnaire (SGRQ) and the Short Form 36-item (SF-36) questionnaires. Linear regression analysis was used to determine the influence of the number or type of medication on the total SGRQ score, adjusting by disease severity and other relevant variables. RESULTS: Significant differences were observed among the number of drugs prescribed according to dyspnea levels, percentage of predicted FEV1 (FEV1%), SGRQ scores and some areas of SF-36. Fifty nine percent of patients with an FEV1%>50% were prescribed inhaled corticosteroids (ICS). Those who took an ICS had a worse HRQL than patients with an FEV1%>50% who did not receive ICS. CONCLUSIONS: A relationship exists between the number of medicines prescribed to patients with COPD and their HRQL, measured by the total SGRQ score, after adjustment by severity of the disease. Within the group of patients who should not have been prescribed ICS, there are subgroups that might benefit from this medication. PMID- 16039841 TI - Anti-tuberculosis treatment and infliximab. PMID- 16039842 TI - Photoelectric properties of a detector based on dried bacteriorhodopsin film. AB - The photoelectric response of a detector using dried bacteriorhodopsin (bR) film as the light sensing material is mathematically modeled and experimentally verified in this paper. The photocycle and proton transfer kinetics of dried bR film differ dramatically from the more commonly studied aqueous bR material because of the dehydration process. The photoelectric response of the dried film is generated by charge displacement and recombination instead of transferring a proton from the cytoplasmic side to the extracellular side of the cell membrane. In this work, the wild-type bR samples are electrophoretically deposited onto an indium tin oxide (ITO) electrode to construct a simple multiple layered photo detector with high sensitivity to small changes in incident illumination. The light absorption characteristics of the thin bR film are mathematically represented using the kinetics of the bR photocycle and the charge displacement theorem. An electrically equivalent RC circuit is used to describe the intrinsic photoelectric properties of the film and external measurement circuitry to analyze the detector's response characteristics. Simulated studies and experimental results show that the resistance of the dried bR film is in the order of 10(11) Omega. When the input impedance of the measurement circuitry is one order of magnitude smaller than the dried film, the detector exhibits a strong differential response to the original time-varying light signal. An analytical solution of the equivalent circuit also reveals that the resistance and capacitance values exhibited by the dried bR film, in the absence of incident light, are almost twice as large as the values obtained while the material is under direct illumination. Experimental observations and a predictive model both support the notion that dried bR film can be used in simple highly sensitive photo-detector designs. PMID- 16039844 TI - Properties of docosahexaenoic-acid-containing lipids and their influence on the function of rhodopsin. AB - The importance of highly polyunsaturated fatty acids in health and development has been convincingly demonstrated by many studies over the past several decades. The mechanisms by which polyunsaturated lipid species might influence biological function at the molecular level are now attracting considerable attention. The G protein-coupled receptor rhodopsin and docosahexaenoic acid, the dominant fatty acid in the retinal membrane, provide the best-studied example of protein function being influenced by lipid environment. PMID- 16039843 TI - TonB-dependent outer membrane transport: going for Baroque? AB - The import of essential organometallic micronutrients (such as iron-siderophores and vitamin B(12)) across the outer membrane of Gram-negative bacteria proceeds via TonB-dependent outer membrane transporters (TBDTs). The TBDT couples to the TonB protein, which is part of a multiprotein complex in the plasma (inner) membrane. Five crystal structures of TBDTs illustrate clearly the architecture of the protein in energy-independent substrate-free and substrate-bound states. In each of the TBDT structures, an N-terminal hatch (or plug or cork) domain occludes the lumen of a 22-stranded beta barrel. The manner by which substrate passes through the transporter (the "hatch-barrel problem") is currently unknown. Solution NMR and X-ray crystallographic structures of various TonB domains indicate a striking structural plasticity of this protein. Thermodynamic, biochemical and bacteriological studies of TonB and TBDTs indicate further that existing structures do not yet capture critical energy-dependent and in vivo conformations of the transport cycle. The reconciliation of structural and non structural experimental data, and the unambiguous experimental elucidation of a detailed molecular mechanism of transport are current challenges for this field. PMID- 16039845 TI - Structure-activity relationships of 2-, 4-, or 6-substituted estrogens as aromatase inhibitors. AB - Aromatase, which is responsible for the conversion of androgens to estrogens, is a potential therapeutic target for the selective lowering of estrogen levels in patients with estrogen-dependent breast cancer. To develop a novel class of aromatase inhibitors, we tested series of 2- and 4-substituted (halogeno, methyl, formyl, methoxy, nitro, and amino) estrones (7 and 9), as well as series of 6alpha- and 6beta-substituted (alkyl, phenalkyl, and alkoxy) estrones (13 and 14), and their estradiol analogs (8, 10, 11, and 12) as aromatase inhibitors. All of the inhibitors examined blocked the androstenedione aromatization in a competitive manner. Introduction of halogeno and methyl functions at C-2 of estrone as well as that of a phenalkyl or methyl function at the C-6alpha or C 6beta position markedly increased affinity to aromatase (apparent K(i) value=0.10 0.66 microM for the inhibitors versus 2.5 microM for estrone). 6alpha Phenylestrone (13c) was the most powerful inhibitor among the estrogens studied, and its affinity was comparable to that of the androgen substrate androstenedione. Estradiol analogs were much weaker inhibitors than the corresponding estrone compounds in each series, indicating that the 17-carbonyl group plays a critical role in the formation of a thermodynamically stable enzyme inhibitor complex. PMID- 16039846 TI - Vitamin D and skin cancer: a problem in gene regulation. AB - The skin is the major source of Vitamin D(3) (cholecalciferol), and ultraviolet light (UV) is critical for its formation. Keratinocytes, the major cell in the epidermis, can further convert Vitamin D(3) to its hormonal form, 1,25 dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] (calcitriol). 1,25(OH)(2)D(3) in turn stimulates the differentiation of keratinocytes, raising the hope that 1,25(OH)(2)D(3) may prevent the development of malignancies in these cells. Skin cancers (squamous cell carcinoma (SCC), basal cell carcinoma (BCC), and melanomas) are the most common cancers afflicting humans. UV exposure is linked to the incidence of these cancers-UV is thus good and bad for epidermal health. Our focus is on the mechanisms by which 1,25(OH)(2)D(3) regulates the differentiation of keratinocytes, and how this regulation breaks down in transformed cells. Skin cancers produce 1,25(OH)(2)D(3), contain ample amounts of the Vitamin D receptor (VDR), and respond to 1,25(OH)(2)D(3) with respect to induction of the 24-hydroxylase, but fail to differentiate in response to 1,25(OH)(2)D(3). Why not? The explanation may lie in the overexpression of the DRIP complex, which by interfering with the normal transition from DRIP to SRC as coactivators of the VDR during differentiation, block the induction of genes required for 1,25(OH)(2)D(3)-induced differentiation. PMID- 16039847 TI - Rodent StAR mRNA is substantially regulated by control of mRNA stability through sites in the 3'-untranslated region and through coupling to ongoing transcription. AB - The steroidogenic acute regulator (StAR) gene is transcribed to 1.6 kb and 3.5 kb mRNAs that differ only through the length of the 3'-untranslated region (3'-UTR). These forms result from alternative polyadenylation sites in exon 7. These sites are utilized similarly in unstimulated adrenal cells whereas Br-cAMP selectively stimulates 3.5 kb mRNA. After removal of Br-cAMP, 3.5 kb mRNA declines rapidly (t(1/2) = 2 h) while 1.6 kb mRNA responds more slowly. This selective degradation is more evident in testis MA10 cells and is seen even in the presence of Br-cAMP. Transfection of Y-1 cells with CMV promoted StAR vectors confirmed that the 3.5 kb form is less stable and that Br-cAMP slowly increases this instability. Basal instability resides solely in the extended 3'-UTR which contains AU-rich elements. Br-cAMP enhances this degradation of 3.5 kb mRNA but additionally requires translated and 5'-UTR sequences. Degradation of both forms is arrested by inhibitors of transcription or translation, indicating that mRNA stability is coupled to these processes independent of the extended 3'-UTR. Br-cAMP stimulates substantial selective synthesis of 3.5 kb StAR mRNA despite this instability. The preferential generation of the unstable form may facilitate rapid increases and decreases of StAR activity in response to external changes. PMID- 16039848 TI - Carbonic anhydrase inhibitors: inhibition of the transmembrane isozyme XIV with sulfonamides. AB - The inhibition of the last human carbonic anhydrase (CA, EC 4.2.1.1) isozyme (hCA XIV) discovered has been investigated with a series of sulfonamides, including some clinically used derivatives (acetazolamide, methazolamide, ethoxzolamide, dichlorophenamide, dorzolamide, brinzolamide, benzolamide, and zonisamide), as well as the sulfamate antiepileptic drug topiramate. The full-length hCA XIV is an enzyme showing a medium-low catalytic activity, quite similar to that of hCA XII, with the following kinetic parameters at 20 degrees C and pH 7.5, for the CO2 hydration reaction: k(cat) = 3.12 x 10(5) s(-1) and k(cat)/K(M) = 3.9 x 10(7) M(-1) s(-1). All types of activities have been detected for the investigated compounds, with several micromolar inhibitors, including zonisamide, topiramate, and simple sulfanilamide derivatives (K(I)-s in the range of 1.46-6.50 microM). In addition, topiramate and zonisamide were observed to behave as weak hCA XII inhibitors, while zonisamide was an effective hCA IX inhibitor (K(I) of 5.1 nM). Some benzene-1,3-disulfonamide derivatives or simple five-membered heteroaromatic sulfonamides showed K(I)-s in the range of 180-680 nM against hCA XIV, whereas the most effective of such inhibitors, including 3-chloro-/bromo-sulfanilamide, benzolamide-like, ethoxzolamide-like, and acetazolamide/methazolamide-like derivatives, showed inhibition constant in the range of 13-48 nM. The best hCA XIV inhibitor was aminobenzolamide (K(I) of 13 nM), but no CA XIV-selective derivatives were evidenced. There are important differences of affinity of these sulfonamides/sulfamates for the three transmembrane CA isozymes, with CA XII showing the highest affinity, followed by CA IX, whereas CA XIV usually showed the lowest affinity for these inhibitors. PMID- 16039849 TI - Epibatidine analogues as selective ligands for the alpha(x)beta2-containing subtypes of nicotinic acetylcholine receptors. AB - A series of epibatidine analogues was synthesized and characterized in vitro. These compounds are high affinity ligands for the nicotinic acetylcholine receptors (nAChR). They display binding selectivity for the alpha(x)beta2 subtypes of nAChRs over the alpha(x)beta4 subtypes, and especially for the alpha4beta2 and alpha2beta2 subtypes. Furthermore, most of these new nicotinic compounds display little, if any, agonist activities at alpha3beta4 nAChR. As a result they might become lead structures for the design and synthesis of highly selective ligands for nAChR subtypes containing the beta2 subunit. PMID- 16039850 TI - Selective cytotoxicity of azatyrosinamides against ras-transformed NIH 3T3 cells. AB - This study aims to develop novel azatyrosinamide compounds structurally modified from ras-specific antioncogenic azatyrosine. Analogues 4-15 were prepared and their inhibition on the growth of wild-type and ras-transformed NIH 3T3 cell lines was compared. Compound 12 was found to be the most active with IC50 16.5+/ 2.2 microM which is 458-fold more potent than that of azatyrosine. The selective toxicity, defined as IC(50 wild-type)/IC(50 ras-transformed) for this compound was 138.5. PMID- 16039851 TI - 8-Piperazinyl-2,3-dihydropyrrolo[3,2-g]isoquinolines: potent, selective, orally bioavailable 5-HT1 receptor ligands. AB - The novel 8-piperazinyl-2,3-dihydropyrroloisoquinoline template was synthesized in nine steps. The template was N-substituted to give a series of compounds showing binding to human cloned 5-HT1A, 5-HT1B and 5-HT1D receptors with pKi's greater than 9 and selectivities up to 1000-fold against other serotonin, dopamine and adrenergic receptors. Several compounds were shown to possess weak partial agonist activity in cloned receptors, which translated to antagonism in in vitro studies. PMID- 16039852 TI - Alpha-rhamnosidase inhibitory activities of polyhydroxylated pyrrolidine. AB - We designed and synthesized polyhydroxylated pyrrolidines 1-12 from L-tyrosine, L phenylalanine, and D-tyrosine through iodine-mediated intramolecular cyclization followed by Woodward-Prevost reaction. The synthetic polyhydroxylated pyrrolidines were identified with structure-based inhibitory activity and selective inhibitory activity against alpha-rhamnosidase. (2S,3S,4R)-deacetyl anisomycin 7 was the best inhibitor among the 12 polyhydroxylated pyrrolidines because it possesses the same stereoconfiguration at C1, C2, C3 as alpha-L rhamnopyranoside. An investigation into the nature of the inhibition showed that the synthetic pyrrolidines are competitive inhibitors. They also did not have remarkable inhibitory activity against seven glycosidases (alpha-glucosidase, alpha-mannosidase, alpha-amylase, beta-glucosidase, beta-galactosidase, beta amylase, and invertase). PMID- 16039853 TI - Carbonic anhydrase inhibitors: design of thioureido sulfonamides with potent isozyme II and XII inhibitory properties and intraocular pressure lowering activity in a rabbit model of glaucoma. AB - A new series of thioureido-substituted sulfonamides were prepared by reacting 4 isothiocyanato- or 4-isothiocyanatoethyl-benzenesulfonamide with amines, hydrazines, or amino acids bearing moieties that can lead to an enhanced hydrosolubility, such as 2-dimethylamino-ethylamine, fluorine-containing aromatic amines/hydrazines, an aminodiol, heterocyclic polyamines (derivatives of morpholine and piperazine), 4-aminobenzoic acid, or natural amino acids (Gly, Cys, Asn, Arg, and Phe). The new compounds showed good inhibitory properties against three physiologically relevant carbonic anhydrase (CA, EC 4.2.1.1) isozymes, with K(I)s in the range of 24-324 nM against the cytosolic isoform CA I, of 6-185 nM against the other cytosolic isozyme CA II, and of 1.5-144 nM against the transmembrane isozyme CA XII. Some of the new derivatives were also very effective in reducing elevated intraocular pressure in hypertensive rabbits as a glaucoma animal model. Considering that this is the first study in which potent CA II/CA XII inhibitors are designed and investigated in vivo, it may be assumed that the target isozymes of the antiglaucoma sulfonamides are indeed the cytosolic CA II and the transmembrane CA XII. PMID- 16039854 TI - 3-(2-Aminoethyl)pyridine analogs as alpha4beta2 nicotinic cholinergic receptor ligands. AB - An examination of several 3-(2-aminoethyl)pyridine analogs suggests that they likely orient at alpha4beta2 nicotinic cholinergic receptors in a different fashion than their correspondingly substituted nicotine analogs. PMID- 16039855 TI - Dipyridyl amines: potent metabotropic glutamate subtype 5 receptor antagonists. AB - Modulation of the metabotropic glutamate subtype 5 (mGlu5) receptor may be useful in the treatment of a variety of central nervous system disorders. Herein, we report on the discovery, synthesis, and biological evaluation of dipyridyl amines as small molecule mGlu5 antagonists. PMID- 16039856 TI - Adamantyl triazoles as selective inhibitors of 11beta-hydroxysteroid dehydrogenase type 1. AB - Adamantyl triazoles were identified as selective inhibitors of 11beta hydroxysteroid dehydrogenase type 1 (11beta-HSD1). They are active both in in vitro and in in vivo pharmacodynamic models. The synthesis and structure-activity relationships of these inhibitors are presented. PMID- 16039857 TI - Coordination of pelvis-HAT (head, arms and trunk) in anterior-posterior and medio lateral directions during treadmill gait in preadolescents with/without Down syndrome. AB - In human biped gait, movements in the frontal plane such as side-to-side rocking, are as essential as the alternating movement of the legs in the sagittal plane. In addition, the top-heavy structure of human body necessitates control of the trunk during walking. In this study, we evaluated the pelvis and HAT (head, arms and trunk) movements and their coordination during treadmill walking in the anterior-posterior and medio-lateral directions in children with typical development (TD) and those with Down syndrome (DS). Participants were 12 children with DS aged 8-10 years and 10 age-matched children with TD. They walked on a treadmill at 40%, 75% and 110% of their preferred overground walking speeds. Kinematic data were collected using a 3D-motion-capture system; movements of the mid-point of hip joints (OPELVIS) and the center of mass of HAT (COMHAT) were reduced. Children with DS showed larger and speed dependent amplitude responses compared to their TD peers. Coordination patterns for children with DS were less stable, especially in medio-lateral direction at slow speed. Differences in amplitude response may be the result of poorer trunk control in children with DS or, alternatively, part of a necessary and sufficient propulsion/stabilization mechanism for this population with reduced tone and muscle strength. Response differences observed between the anterior-posterior and medio-lateral directions for both groups may reflect relative differences in the involvement of active neuromuscular control. PMID- 16039858 TI - Taking metagenomic studies in context. PMID- 16039859 TI - Synthesis and antimycobacterial evaluation of various 7-substituted ciprofloxacin derivatives. AB - Tuberculosis continues to be a major cause of morbidity and mortality all over the world. Various 7-substituted ciprofloxacin derivatives were synthesized and evaluated for antimycobacterial activity in vitro and in vivo against Mycobacterium tuberculosis and for inhibition of the supercoiling activity of DNA gyrase from Mycobacterium smegmatis. Preliminary results indicated that most of the compounds demonstrated better in vitro antimycobacterial activity against M. tuberculosis than ciprofloxacin. Compound 1-cyclopropyl-6-fluoro-1,4-dihydro-4 oxo-7-[[N4-[1'-(5-methylisatinyl-beta-semicarbazo)]methyl]N1-piperazinyl]-3 quinoline carboxylic acid (3h) decreased the bacterial load in spleen tissue with 0.76-log10 protections and was considered to be moderately active in reducing bacterial count in spleen. The results demonstrated the potential and importance of developing new quinolone derivatives against mycobacterial infections. PMID- 16039860 TI - Irreversibly inhibitory kinetics of 3,5-dihydroxyphenyl decanoate on mushroom (Agaricus bisporus) tyrosinase. AB - 3,5-Dihydroxyphenyl decanoate (DPD) is found to inhibit the diphenolase activity of tyrosinase from mushroom (Agaricus bisporus). The effects of DPD on the diphenolase activity of mushroom tyrosinase have been studied. The results show that the enzyme activity decreases very slowly with an increase in DPD concentrations at lower concentrations of DPD (between 5 and 60 microM). But at higher concentrations of DPD, DPD can strongly inhibit the diphenolase activity of the enzyme and the inhibition is irreversible. The IC50 value was estimated to be 96.5 microM. The inhibition mechanism of DPD has been investigated and the results show that DPD can bind to the free enzyme molecule and enzyme-substrate complex and lose the enzyme activity completely. The inhibition kinetics has been studied in detail by using the kinetic method of the substrate reaction described by Tsou. The microscopic rate constants of the enzyme inhibited by DPD at higher concentrations have been determined. PMID- 16039861 TI - Synthesis and evaluation of molluscicidal and larvicidal activities of some novel enaminones derived from 4-hydroxyquinolinones: part IX. AB - A series of 10 3-(hetarylaminomethylene)quinolinediones, 12 3-(substituted aminopropenoyl)-4-hydroxyquinolinones, and 10 3-(substituted aminomethylene-5-oxo pyrazolinyl)-4-hydroxyquinolinones were synthesized as novel enaminones derived from 3-(un)substituted 4-hydroxyquinolin-2(1H)-ones in 72-94% yields and assayed for their molluscicidal activities against Biomphalaria alexandrina and Lymnaea natalensis snails. Some of the tested enaminones presented high molluscicidal activities (LC(50)20ppm). The new compounds showed more potency against hatchability of B. alexandrina egg masses, the infection rate and prepatent period of the snails. In addition, these derivatives revealed potential larvicidal effects (100% mortality) on both miracidia and cercariae of Schistosoma mansoni at reduced exposure time. The selected active derivatives were examined against Daphnia magna and their nontoxic effect at all sublethal, lethal, and higher concentrations suggests that these compounds can play an important role as molluscicides and larvicides with environmental safe properties. PMID- 16039862 TI - Synthesis of novel substituted 2-phenylpyrazolopyridines with potent activity against herpesviruses. AB - Herpesviruses are a significant source of human disease; amongst these herpes simplex virus 1 (HSV-1) and HSV-2 are very prevalent and cause recurrent infections. We recently identified a pyrazolo[1,5-a]pyridine scaffold that showed promising activity against HSV-1 and HSV-2 in Vero cell antiviral assays. Here, we describe the synthesis and anti-herpetic activity of several 3-pyrimidinyl-2 phenylpyrazolo[1,5-a]pyridines with differing 2-phenyl substitution patterns. Approaches to rapidly access a number of analogs with different 2-phenyl substitution patterns are outlined. Several of the compounds described have comparable activity to acyclovir against HSV-1 and HSV-2. PMID- 16039863 TI - Novel 5-substituted, 2,4-diaminofuro[2,3-d]pyrimidines as multireceptor tyrosine kinase and dihydrofolate reductase inhibitors with antiangiogenic and antitumor activity. AB - Recent evidence suggests that combination therapy of cancer with receptor tyrosine kinase (RTK) inhibitors, which are usually cytostatic, with conventional chemotherapeutic agents, which are usually cytotoxic, provide an improved treatment option. We have designed, synthesized, and evaluated a series of novel 2,4-diamino-5-substituted furo[2,3-d]pyrimidines with RTK and dihydrofolate reductase (DHFR) inhibitory activity in single molecules, as potential cytostatic and cytotoxic agents with antitumor activity. These compounds were synthesized from 2,4-diamino-5-chloromethyl furo[2,3-d]pyrimidine and aryl methyl ketones using the Wittig reaction to afford the C-8-C-9 unsaturated analogs followed by catalytic reduction to the corresponding saturated compounds. The saturated and unsaturated C-8-C-9 bridged compounds were evaluated as inhibitors of vascular endothelial growth factor receptor (VEGFR-2, Flk, KDR), epidermal growth factor receptor, and platelet-derived growth factor receptor-beta (PDGFR-beta). Selected analogs were also evaluated as antiangiogenic agents in the chicken embryo chorioallantoic membrane (CAM) assay. The compounds were also evaluated as inhibitors of human (h) DHFR and Toxoplasma gondii (tg) DHFR. In each evaluation, a known standard compound was used as a comparison. Of the compounds evaluated, compound 32 was as potent as the standard compounds against VEGFR-2 and PDGFR beta, showing dual inhibitory activity against RTK. This analog was also highly effective in the CAM assay. A second analog 18 also demonstrated dual VEGFR-2 and PDGFR-beta inhibitory activity as well as potent antiangiogenic activity in the CAM assay. Four additional analogs were also effective against PDGFR-beta and in the CAM assay. An unsaturated C-8-C-9 moiety was necessary for RTK inhibitory activity. Compound 32 also showed inhibitory activity against hDHFR and tgDHFR, illustrating the multitarget inhibitory potential of these analogs. The biological activity of these analogs also suggests the necessity of an unsaturated C-8-C-9 bridge for dual RTK and DHFR inhibitory activity. Compounds 18 and 32 were also evaluated in a B16 melanoma mouse model and were found to be more active as antitumor agents than methotrexate. In addition, both 18 and 32 were also active in decreasing lung metastases in a mouse model of B16 melanomas. PMID- 16039864 TI - A feasibility study of a solid-state microdosimeter. AB - A solid-state silicon detector is a challenging device for microdosimetry, mainly because it can provide sensitive zones of the order of a micrometer. Moreover, these detectors are characterized by a high spatial and a good energy resolution. However, they may present some limitations, such as: (i) the minimum detectable energy which is limited by the electronic noise; (ii) radiation hardness; (iii) the geometry of the sensitive volume; (iv) the field-funnelling effect; (v) the non-tissue-equivalence of silicon. This work discusses a feasibility study of a microdosimeter based on a monolithic silicon telescope, consisting of a DeltaE and an E stage-detector, about 1 and 500 microm thick, respectively. Charges are collected separately in the two stage-detectors. The use of the DeltaE stage coupled with a tissue-equivalent converter was investigated as a solid-state microdosimeter. Irradiations with monoenergetic neutrons were performed at the INFN-Laboratori Nazionali di Legnaro (Italy). The field-funnelling effect appears to be negligible from the comparison of the experimental data with the results of Monte Carlo simulations, performed with the FLUKA code. The preliminary results of an analytical approach for the correction for geometrical effects and tissue equivalence are also presented. PMID- 16039865 TI - Comparisons of activity measurements with radionuclide calibrators-A tool for quality assessment and improvement in nuclear medicine. AB - A national program of ongoing comparisons for assaying gamma-emitting radiopharmaceuticals for amount of radioactivity using radionuclide calibrators was begun in 2000. Nuclides of the most wide-spread use in Cuban nuclear medicine, 131I, 201Tl and (99m)Tc, as well as two measurement geometries, glass vials and plastic syringes, were employed. In this paper, the participants' performance is assessed by mean of a statistical analysis of the reported data. Performance tables have been obtained and a chi2 statistic is calculated from observed and expected frequencies, with the aim of testing the hypothesis about the independence of some characteristics of the comparison results, at a significance level alpha=0.05. The proportion of satisfactory results in the years 2002-2004 were found to be at the same level, but higher than in 2000. It reveals an improvement of the measurement quality since 2002. The causes of improvement were investigated using the statistical treatment of several data available as supplementary information. PMID- 16039866 TI - Blood-brain barrier alterations in both focal and diffuse abnormalities on postmortem MRI in multiple sclerosis. AB - Postmortem MRI-guided tissue sampling significantly enhances the yield of MS lesions in autopsy material, but so far it is unknown whether abnormalities concur with blood-brain barrier alterations. Here we sampled MS lesions with focal and diffuse abnormalities (diffusely abnormal white matter; DAWM) on MRI; both were coupled to the presence of MS lesions upon neuropathological examination. Extravascular distribution of fibrinogen, indicating BBB disturbance, was observed in so-called (p)reactive lesions that reflect discrete areas of microglial activation without demyelination within an otherwise normal appearing white matter. Leakage became more extensive in active demyelinating MS lesions to chronic inactive lesions. An enlargement of the perivascular (Virchow Robin) space containing infiltrated leukocytes was associated with both DAWM and focal abnormalities on postmortem MRI. This study shows for the first time that in MS brain changes in the vasculature take place not only in focal lesions but also in DAWM as detected by postmortem MRI. PMID- 16039867 TI - Taxanes in adjuvant breast cancer setting: which standard in Europe? AB - The clinical studies of cooperators groups (trials CALGB 9344, NSABP-B-28, BCIRG 001, PACS01 and CALGB 9741) demonstrated, in the adjuvant breast cancer setting, that taxanes (paclitaxel and docetaxel) improved both disease free and overall survival (trials CALGB 9344, BCIRG 001 and PACS 01). However, the debate remains open in Europe. Less than 50% of the expert present at the last St. Gallen Conference recommended the use of taxanes in adjuvant setting. The reasons for this are primarily related to the fact that the comparator arms of cooperators group (AC, FAC and FEC 100) are considered by some Europe groups as being less effective than the European standards (chemotherapy (CMF) and Epirubicine-CMF). Many questions remain unanswered, including whether the use of taxanes should be sequential or concomitant, and which population would benefit from such a treatment: patients with hormone-receptor negative disease and/or the HER-2 positive tumors? PMID- 16039868 TI - Rapamycin: an anti-cancer immunosuppressant? AB - Rapamycin and its derivatives are promising therapeutic agents with both immunosuppressant and anti-tumor properties. These rapamycin actions are mediated through the specific inhibition of the mTOR protein kinase. mTOR serves as part of an evolutionarily conserved signaling pathway that controls the cell cycle in response to changing nutrient levels. The mTOR signaling network contains a number of tumor suppressor genes including PTEN, LKB1, TSC1, and TSC2, and a number of proto-oncogenes including PI3K, Akt, and eIF4E, and mTOR signaling is constitutively activated in many tumor types. These observations point to mTOR as an ideal target for anti-cancer agents and suggest that rapamycin is such an agent. In fact, early preclinical and clinical studies indicate that rapamycin derivatives have efficacy as anti-tumor agents both alone, and when combined with other modes of therapy. Rapamycin appears to inhibit tumor growth by halting tumor cell proliferation, inducing tumor cell apoptosis, and suppressing tumor angiogenesis. Rapamycin immunosuppressant actions result from the inhibition of T and B cell proliferation through the same mechanisms that rapamycin blocks cancer cell proliferation. Therefore, one might think that rapamycin-induced immunosuppression would be detrimental to the use of rapamycin as an anti-cancer agent. To the contrary, rapamycin decreases the frequency of tumor formation that occurs in organ transplant experiments when combined with the widely used immunosuppressant cyclosporine compared with the tumor incidence observed when cyclosporine is used alone. The available evidence indicates that with respect to tumor growth, rapamycin anti-cancer activities are dominant over rapamycin immunosuppressant effects. PMID- 16039869 TI - Immunosuppressive agents in solid organ transplantation: Mechanisms of action and therapeutic efficacy. AB - Effective immunosuppression is an essential pre-requisite for successful organ transplantation and improvements in outcome after transplantation have to a large extent been dependent on developments in immunosuppressive therapy. Here we provide an overview of the different immunosuppressive agents currently used in solid organ transplantation. A historical perspective on the development of immunosuppression for organ transplantation is followed by a review of the individual agents, with a focus on their mechanism of action and efficacy. Steroids, anti-proliferative agents (azathioprine and mycophenolate), calcineurin inhibitors (cyclosporine and tacrolimus) and TOR inhibitors (sirolimus and everolimus) are discussed along with both polyclonal and monoclonal antibody preparations. Many of the key clinical trials that underpin current clinical usage of these agents are described and side-effects of the different agents are highlighted. Finally, a number of newer agents still in various stages of clinical development are briefly considered. PMID- 16039870 TI - Multistep skin cancer in mice as a model to study the evolution of cancer cells. AB - Although much of cancer research relies on Nowell's clonal evolution hypothesis as a conceptual framework, large gaps remain in understanding how tumors develop. The multistage skin cancer model in mice provides continuing insight on fundamental aspects of tumor evolution. In this model, mutation of the oncogene Hras is frequently the initiating event while mutation of the tumor suppressor p53 is a late event, associated with malignant progression. Recent evidence demonstrates that intracellular signaling from the initial Hras mutation leads directly to the activation of p53, creating selective pressure in favor of cells with mutant p53. Thus, selection for subsequent mutations is mechanistically linked to the initial mutation, explaining the preferred order of mutational events observed. Analysis of this model also reveals that a diverse array of signals can selectively impair or enhance clonal expansion of Ras mutant cells into a visible neoplasm. These modifiers can be genetic, physiological, or environmental and are often highly specific to tumor cells. This indicates that tumor cells have an inherent reduced capacity to buffer against perturbations. Reduced buffering may play an important role in both tumor evolution and therapy response and may be a hallmark of cancer cells. PMID- 16039871 TI - Disrupted Schwann cell-axon interactions in peripheral nerves of mice with altered L1-integrin interactions. AB - The cell adhesion molecule L1 is important for peripheral nerve development. Mice lacking the 6th Ig domain of L1 (L1-6D mice) lose L1 homophilic binding and RGD dependent L1-integrin binding [Itoh, K., Cheng, L., Kamei, Y., Fushiki, S., Kamiguchi, H., Gutwein, P., Stoeck, A., Arnold, B., Altevogt, P., Lemmon, V., 2004. Brain development in mice lacking L1-L1 homophilic adhesion. J. Cell Biol. 165, 145-154]. We examined the ultrastructure of sciatic nerves from L1-6D at postnatal day 7 and 8 weeks. Unmyelinated axons frequently detached at the edge of Schwann cells, and naked axons were observed. Myelin was thinner in L1-6D and abnormal, multiple axons wrapped in a single myelin sheath were routinely observed. Previous work has shown that L1 on axons interacts with a heterophilic binding partner on Schwann cells to facilitate normal peripheral nerve formation. Taken together, it is likely that L1 on axons binds integrins on Schwann cells, resulting in interactions between axons and Schwann cells that are essential for ensheathment and myelination. PMID- 16039872 TI - In vitro activation, purification, and characterization of Escherichia coli expressed aryl-alcohol oxidase, a unique H2O2-producing enzyme. AB - Aryl-alcohol oxidase (AAO), a flavoenzyme with unique spectral and catalytic properties that provides H2O2 for fungal degradation of lignin, has been successfully activated in vitro after Escherichia coli expression. The recombinant AAO (AAO*) protein was recovered from inclusion bodies of E. coli W3110 transformed with pFLAG1 containing the aao cDNA from Pleurotus eryngii. Optimization of in vitro refolding yielded 75% active enzyme after incubation of AAO* protein (10 microg/ml) for 80 h (at 16 degrees C and pH 9) in the presence of glycerol (35%), urea (0.6 M), glutathione (GSSG/GSH molar ratio of 2), and FAD (0.08 mM). For large-scale production, the refolding volume was 15-fold reduced and over 45 mg of pure active AAO* was obtained per liter of E. coli culture after a single anion-exchange chromatographic step. Correct FAD binding and enzyme conformation were verified by UV-visible spectroscopy and circular dichroism. Although the three enzymes oxidized the same aromatic and aliphatic polyunsaturated primary alcohols, some differences in physicochemical properties, including lower pH and thermal stability, were observed when the activated enzyme was compared with fungal AAO from P. eryngii (wild enzyme) and Emericella nidulans (recombinant enzyme), which are probably related to the absence of glycosylation in the E. coli expressed AAO. PMID- 16039873 TI - The relation of postmenopausal hormone therapy to serum uric acid and the risk of coronary heart disease events: the Heart and Estrogen-Progestin Replacement Study (HERS). AB - PURPOSE: To determine whether baseline serum uric acid (UA) levels and estrogen progestin (E+P)-associated change in serum UA in postmenopausal women with coronary disease are associated with recurrent coronary heart disease (CHD) events. METHODS: 2763 postmenopausal women enrolled in the Heart and Estrogen Progestin Replacement Study (HERS) were randomly assigned to take conjugated E+P or placebo in a secondary CHD prevention study. The primary outcome for these analyses was nonfatal myocardial infarction or CHD death during a mean follow up of 4.1 years. RESULTS: The baseline serum UA for the cohort was 5.4 mg/dl and, compared with placebo, E+P on average lowered serum UA levels slightly (0.2 mg/dl) at one year of follow up (p<0.0001). Baseline serum UA levels were associated in simple proportional hazards models with CHD events; each standard deviation increase (1.3 mg/dl) was associated with a 22% increased risk of primary CHD events (p=.0001). This association, however, was no longer statistically significant after multivariable adjustment (p=0.36). There was no association between on-study change in serum UA level and any CHD outcome. CONCLUSION: Treatment with E+P lowered serum UA levels slightly, but neither baseline UA nor change in UA affected CHD risk. PMID- 16039874 TI - Sex differences in the association between birth weight and total cholesterol. A meta-analysis. AB - PURPOSE: Determine whether a sex difference exists in the association between birth weight and total cholesterol later in life. METHODS: Meta-analysis of within-study differences in regression coefficients of cholesterol on birth weight. RESULTS: A total of 34 regression coefficients from 30 studies were included in the analyses; these provided data on 33,650 males and 23,129 females. There was evidence that the inverse association between birth weight and total cholesterol was stronger in males compared to females. The pooled within-study difference in age-adjusted regression coefficients was -0.03 mmol/l (-0.06, 0.01), p = 0.02 and the pooled within-study difference in age and body mass index adjusted regression coefficients was -0.04 mmol/l (-0.07, -0.02), p = 0.002. There was no evidence of heterogeneity in these meta-analyses (both p values > 0.6). CONCLUSIONS: These results provide some evidence for a sex difference in the birth weight-total cholesterol association. This is consistent with studies of fetal growth which suggest that birth size reflects different biological processes for females and males. However, other very large studies are required to confirm this finding. PMID- 16039875 TI - Reliability of Indiana birth certificate data compared to medical records. AB - PURPOSE: The purpose of this study was to measure the reliability of data reported on Indiana electronic birth certificates. Knowing the accuracy of birth certificate data is crucial when identifying community health needs and evaluating birth outcomes interventions. METHODS: This study compared 1996 electronic birth certificate data on a random sample of 1050 Indiana hospital births to data abstracted from the hospital medical records for the same patients. Kappa scores, Pearson r correlation values, sensitivity, specificity, and positive predictive values of the birth certificate data were used to measure agreement. RESULTS: Parents' demographic variables had the best agreement, followed by birth outcome variables. Delivery type, cesarean indications, pregnancy history, prenatal care and mother's risk variables were found to have moderate agreement. Agreement was poor for variables measuring labor and delivery complications, obstetric procedures, concurrent illnesses, pregnancy complications, congenital anomalies, and abnormal conditions. CONCLUSIONS: The results of this study clearly show that some important descriptive and outcome data are reliable while infrequent events are generally not. The results indicate a need to improve the quality of data reported on birth certificates. PMID- 16039876 TI - Weighting response to adjust for nonresponse in SIR analysis in cohort studies. PMID- 16039877 TI - Cardiovascular disease in patients with chronic obstructive pulmonary disease, Saskatchewan Canada cardiovascular disease in COPD patients. AB - PURPOSE: To measure prevalence, incidence, and mortality of cardiovascular outcomes among persons with chronic obstructive pulmonary disease (COPD) and to assess the extent these outcomes differ from persons without COPD. METHODS: Retrospective cohort study in longitudinal health care databases maintained by the government of Saskatchewan, Canada. Subjects were persons age 40 years or older who were diagnosed with COPD during 1997-2000 and who received two or more prescriptions for bronchodilators within 6 months of diagnosis. Each subject was matched by age and sex to two controls without COPD or asthma. RESULTS: Of COPD patients (n = 11,493), 54% were male, and 74% were 65 years or older. Prevalence of all cardiovascular diseases was higher in the COPD group than in the comparison group. After adjusting for cardiovascular risk, odds ratios of prevalence were: arrhythmia 1.76 (confidence interval [CI]: 1.64-1.89), angina 1.61 (CI: 1.47-1.76), acute myocardial infarction 1.61 (CI: 1.43-1.81), congestive heart failure 3.84(CI: 3.56-4.14), stroke 1.11 (CI: 1.02-1.21), pulmonary embolism 5.46 (CI: 4.25-7.02). Risk of hospitalization due to each cardiovascular cause was elevated in the COPD group. The risk ratio for cardiovascular mortality was 2.07 (CI: 1.82-2.36) and all cause mortality was 2.82 (CI: 2.61-3.05). CONCLUSIONS: Persons with diagnosed and treated COPD are at increased risk for hospitalizations and deaths due to cardiovascular diseases. PMID- 16039878 TI - Trajectory of systolic blood pressure in children and adolescents. AB - PURPOSE: Rapid height and weight changes during childhood contribute markedly to blood-pressure change during children's physical growth. This article evaluates the differences in systolic blood pressure (SBP) growth or changes between four gender-ethnic groups: African American males (AM), Caucasian males (CM), African American females (AF), and Caucasian females (CF). METHODS: Subjects 6-9 years old at entry (n = 1302) were followed for 12 years. The repeated-measure data of SBP were analyzed using the Gompertz growth model with random coefficients. RESULTS: Mean SBP (mmHg) at age 6 years was lowest in African American girls (82.23 +/- 0.76) and highest in Caucasian boys (102.83 +/- 0.5). And for both ethnic groups, girls had lower levels at which SBP growth stopped. The peak growth ages (years) also differed by group: 9.30 +/- 0.73, 9.91 +/- 0.28, 10.00 +/- 0.82, and 10.60 +/- 0.22 for African American girls, African American boys, Caucasian girls and Caucasian boys, respectively. CONCLUSION: SBP growth differed among gender-ethnic groups with respect to mean SBP level at age 6, the level at which SBP growth stops and the mean age at which SBP growth rate was at its peak. PMID- 16039879 TI - Complement component C3 transcription in Atlantic halibut (Hippoglossus hippoglossus L.) larvae. AB - The complement systems of fish are well developed and play an important role in the innate immune response. Complement C3 is the central protein of all three activation pathways and is the major opsonin of the complement system and essential for the generation of the membrane attack complex. A 1548 bp part of complement component C3 was isolated from a halibut liver cDNA library by immunoscreening. The deduced amino acid sequence showed that this part of halibut C3 contained key amino acids for factor H, I and properdin binding as well as two N-glycosylation sites. Digoxigenine labelled mRNA probes were synthesised and the transcription of C3 was monitored in three larval stages at 206, 430 and 1000 degrees d (30, 50 and 99 days post hatching), by in situ hybridisation. C3 mRNA was detected in muscle, liver, brain, chondrocytes, spinal cord, eye, intestines, oesophagus and kidney. These findings are in accordance with a former immunohistochemical study on halibut C3 protein ontogeny, indicating that C3 is indeed locally expressed in many organs from the youngest stages on. Complement may thus be linked to the formation and generation of different organs during development and play an important role in the early immune response of halibut larvae. PMID- 16039880 TI - Tissue distribution of prophenoloxidase transcript in the Pacific white shrimp Litopenaeus vannamei. AB - The expression of prophenoloxidase (proPO) transcript in the tissues of the Pacific white shrimp Litopenaeus vannamei was studied by Northern blot, RT-PCR and in situ hybridisation. Northern blot analysis reveals that proPO was constitutive mainly in shrimp haemocytes. Its transcripts, when detected with RT PCR, were widely distributed in haemocytes, gill, heart, lymphoid organ, stomach, midgut, anterior midgut caecum and ganglion. A lower expression was found in hepatopancreas, muscle and cuticular epidermis. The results of in situ hybridisations showed that gill, heart, muscle, haematopoitic tissue and hepatopancreas that were infiltrated by haemocytes also presented positive signals. ProPO transcripts, however, were detected in non-haemocyte cells, including F and E cells of the hepatopancreas, epithelium in stomach, anterior midgut caecum, glia cell in nervous cord and neurosecretory cells in ganglions. PMID- 16039881 TI - Miglustat (NB-DNJ) works as a chaperone for mutated acid beta-glucosidase in cells transfected with several Gaucher disease mutations. AB - Gaucher disease (GD) is a disorder of glycosphinglipid metabolism caused by deficiency of lysosomal acid beta-glucosidase (GC), resulting in progressive deposition of glucosylceramide in macrophages. The glucose analogue, N-butyl deoxynojirimycin (NB-DNJ, Miglustat), is an inhibitor of the ceramide-specific glucosyltransferase (CSG) which catalyzes the first step of glycosphingolipids biosynthesis and is currently approved for the oral treatment of type 1 GD. Using site-directed mutagenesis, we constructed plasmids containing wild-type and several mutations in glucocerebrosidase (GBA) gene. The plasmids were transfected into COS-7 cells and stable transfected cell lines were obtained by geneticin (G418) selection. Cells were cultured during 6 days with medium with or without 10 microM NB-DNJ. The addition of NB-DNJ to COS-7 cell medium leads to 1.3-, 2.1 , 2.3-, 3.6-, and 9.9-fold increase in the activity of S364R, wild-type, N370S, V15M, and M123T GC, respectively. However, no significant changes were observed in the activity of the L444P, L336P, and S465del mutated proteins, but a small decrease in the rare P266L variant was observed. These results suggest that NB DNJ, in addition to the inhibitory effect on CSG, also works as a "chemical chaperone", increasing the activity of acid beta-glucosidase of wild-type and several GC mutated proteins, including the most frequent N370S mutation. The specific location of the Miglustat binding site in GC is unknown. Potential binding sites in the enzyme have been searched for using computational molecular docking. The searching strategy identified three potential GC binding sites for Miglustat, one being the substrate-binding site of the enzyme, which was the best ranked site by AutoDock program. Therefore, it is possible that Miglustat exerts its chaperoning activity on acid beta-glucosidase by acting as an inhibitor bound at the active site. This increase on the activity of the acid beta-glucosidase would imply that Miglustat is not only a substrate reducer but also an inhibitor of the GC degradation, with very promising clinical implications for the treatment of GD patients. PMID- 16039882 TI - Early steps in neural crest specification. AB - The neural crest is a multipotent cell population that arise at the border of the neural plate and non-neural ectoderm. Studies conducted in a number of model organisms including chickens, frogs, zebrafish and mice have been instrumental in elucidating this molecular mechanisms underlying neural crest formation. Signaling molecules of the Wnt, BMP, and FGF families and their downstream effectors have been shown to mediate neural crest induction. Transcription factors including members of the Snail and SoxE gene families as well as FoxD3, c Myc and others have been implicated in specification of the neural crest. These studies represent an important step in understanding the regulatory interactions involved in generating this complex and interesting cell type. PMID- 16039883 TI - Sox proteins and neural crest development. AB - Among the families of transcription factors expressed at the neural plate border in response to neural crest-inducing signals, Sox proteins have emerged as important players in regulating multiple aspects of neural crest development. Here, we summarize the expression of six Sox genes, namely Sox8, Sox9, Sox10, LSox5, Sox4 and Sox11, in neural crest progenitors and their derivatives, and review some aspects of their function pertaining to neural crest development in several species. PMID- 16039884 TI - Optically detected nuclear magnetic resonance at the sub-micron scale. AB - Nuclear magnetic resonance is arguably one of the most powerful techniques available today to characterize diverse systems. However, the low sensitivity of the standard detection method constrains the applicability of this technique to samples having effective dimensions not less than a few microns. Here, we propose a novel scheme and device for the indirect detection of the nuclear spin signal at a submicroscopic scale. This approach--for which the name Dipolar Field Microscopy is suggested--is based on the manipulation of the long-range nuclear dipolar interaction created between the sample and a semiconductor tip located close to its surface. After a preparation interval, the local magnetization of the sample is used to modulate the nuclear magnetization in the semiconductor tip, which, in turn is determined by an optical inspection. Based on results previously reported, it is shown that, in principle, images and/or localized high resolution spectra of the sample can be retrieved with spatial resolution proportional to the size of the tip. PMID- 16039885 TI - Heteronuclear decoupling under fast MAS by a rotor-synchronized Hahn-echo pulse train. AB - A new heteronuclear decoupling mechanism under fast magic-angle spinning MAS is introduced. It is based on refocusing the coherences responsible for the dephase of low-gamma nuclei ((13)C, (15)N) transverse spin-polarization in the presence of strongly dipolar-coupled protons, and has the advantage that can be implemented by pulsed techniques, with all the benefits resulting from a reduced duty cycle compared with conventional decoupling by continuous rf irradiation. The decoupling efficiency of a simple rotor-synchronized Hahn-echo pulse train is analyzed both theoretically and experimentally. It was found that a substantial improvement in sensitivity and resolution can be achieved in compounds with small (1)H chemical shielding parameters even at moderate sample spinning, and some interesting applications are shown. It is also shown that much faster spinning frequencies, or alternative refocusing sequences, are needed for applications on rigid organic solids, i.e., in systems with larger (1)H chemical shifts. PMID- 16039886 TI - 1H-detected double-J-modulated INEPT-INADEQUATE for simultaneous determination of one-bond and long-range carbon-carbon connectivities and the measurement of all carbon-carbon coupling constants. AB - We describe a new NMR experiment, (1)H-detected double-J-modulated (DJM)-INEPT INADEQUATE, for tracing out the carbon skeleton of molecules. This experiment allows simultaneous correlation of directly bonded carbon atoms and those separated by multiple bonds, while at the same time also providing the values of all J(CC) coupling constants. This is achieved by replacing both fixed carbon carbon coupling evolution intervals of the INEPT-INADEQUATE experiment by variable time intervals, which are incremented in concert with the DQ evolution period (t(1)). We show that the analysis of the fine structure of cross-peaks in DJM-INEPT-INADEQUATE spectra leads to accurate values of coupling constants and give guidelines for the proper usage of the method. The proposed experiment is two times less sensitive that the original INEPT-INADEQUATE experiment. We show that, using a 600-MHz cryoprobe and 20 mg of a monosaccharide, spectra that are suitable for the analysis of coupling constants as small as 2 Hz can be obtained within 24 h. Instead of performing multiple experiments, a single DJM-INEPT INADEQUATE experiment can thus provide a wealth of information for the structural analysis of small molecules. PMID- 16039887 TI - Growth hormone treatment of idiopathic short stature: clinical studies. AB - The selection of short-statured children for growth hormone (GH) treatment has long been complicated by the requirement for provocative testing that is unphysiologic, difficult to administer, and potentially dangerous. The recent FDA approval of GH for the treatment of children with idiopathic short stature allows treatment decisions to be based more on the degree of short stature and the potential for attaining a normal adult height. Several studies conducted in children have shown that GH therapy can effectively and safely produce height outcomes within a normal adult range. The observed variability in height response associated with GH use in some of these studies has left the clinician with a difficult decision. Still, the availability of GH for this patient population provides a rational treatment option for patients who fail arbitrary and inaccurate assessments. PMID- 16039889 TI - Renal osteodystrophy: a pediatric perspective, 2005. AB - Renal osteodystrophy (ROD) associated with chronic kidney disease (CKD) involves a complex interrelationship of the loss of divalent mineral homeostasis, hyperparathyroidism, and gene modulation. In affected children, ROD leads to impaired linear growth as well as to the development of other significant skeletal and extraskeletal manifestations. Despite the success of kidney transplantation, many ROD complications cannot be completely reversed; and thus, patients with CKD and ROD require long-term follow-up. Although the availability of vitamin D analogues has advanced ROD management, it has also created new issues for clinicians to address, motivating future investigations of calcimimetic therapies. An algorithmic approach to the management of ROD in children is presented; to be most effective, this approach requires close and frequent surveillance to prevent side effects related to potent therapies. PMID- 16039888 TI - Treatment of the adult growth hormone deficiency syndrome: directions for future research. AB - The adult growth hormone deficiency syndrome (AGHD) is a well-defined clinical entity characterized by decreased lean body mass and bone mineral density, increased visceral adiposity, abnormal lipid profile, decreased muscle strength and exercise endurance, and diminished quality of life. Recent studies have emphasized the increased morbidity and mortality of hypopituitary patients, and there are now data implicating growth hormone (GH) deficiency as a cause of this increase. GH replacement therapy has been shown to reverse many of these abnormalities and to be well tolerated. Meta-analyses have demonstrated that GH treatment positively affects cardiovascular risk factors, and controlled trials have shown that visceral adiposity decreases in treated patients. Improvements in bone mineral density and decreases in fracture rates have also been reported, and new studies using disease-specific questionnaires provide convincing evidence that GH greatly enhances quality of life. Epidemiologic studies indicate that higher insulin-like growth factor I (IGF-I) levels may predict risk for certain cancers, but other studies suggest that lower IGF-I levels increase risk for ischemic heart disease. However, much of the community of endocrine caregivers remains skeptical of GH treatment and, therefore, a large fraction of patients with AGHD are not treated. The accumulation of long-term treatment data will be required to provide reassurance that GH treatment is a safe and necessary form of hormone replacement therapy for patients with AGHD. PMID- 16039890 TI - Neurosurgical approach to treating pituitary adenomas. AB - The transsphenoidal route remains the dominant surgical approach to the management of pituitary adenomas. These tumors may present clinically with signs or symptoms related to hypersecretion, hypopituitarism, or mass effect; or they may be identified incidentally during neuroimaging for management of other disorders. Diagnostic and treatment strategies for pituitary adenomas are reviewed. In addition, we present outcomes data from a large number of patients treated over a 30-year period by a single neurosurgeon. In our experience, transsphenoidal surgery for pituitary adenomas is associated with low rates of morbidity, mortality, and disease recurrence. PMID- 16039891 TI - A review of the effects of therapy on growth and bone mineralization in children with congenital adrenal hyperplasia. AB - The medical management of children with congenital adrenal hyperplasia (CAH) can be challenging with regard to optimizing final height. Insufficient glucocorticoid suppression of adrenal hormone production will result in acceleration of bone maturation and premature epiphyseal fusion, while appropriate or excessive glucocorticoid therapy can be accompanied by suppression of the growth axis. The extent of the growth suppression appears to be affected by the type and dose of glucocorticoid. Some studies suggest that such growth suppression can be ameliorated through concomitant use of growth hormone (GH) therapy. Available data published over the last 10 years on height outcomes in CAH patients treated with glucocorticoids will be reviewed. PMID- 16039892 TI - A brief review of the use and utility of growth hormone stimulation testing in the NCGS: do we need to do provocative GH testing? AB - True growth hormone deficiency (GHD) in childhood, while rare, has major clinical consequences. GHD is often associated with other pituitary hormone deficiencies, so these children may require multiple hormonal replacement and close clinical follow-up to optimize their outcome. Growth hormone stimulation testing (GHST), as currently conducted, is not a reliable diagnostic tool. Both changes in growth hormone assay methodologies and increases in the diagnostic threshold contribute to the incorrect labeling of a substantial proportion of normal children as having idiopathic GHD. Fortunately, newer imaging technologies and laboratory tests form a more rational basis to diagnose true GHD. The use of GHST among GH naive subjects (<20 years of age) enrolled in the National Cooperative Growth Study has declined over the past two decades, from a high of 89% between 1987 and 1989 to only 52% in 2002. Given that GH stimulation testing does not meaningfully aid in distinguishing those few children with true growth hormone deficiency from the much more common short normal child and that alternatives are now available, it is time to discontinue the routine use of GHST in children. PMID- 16039893 TI - Insulin-like growth factor I (IGF-I) measurements in growth hormone (GH) therapy of idiopathic short stature (ISS). AB - Growth hormone (GH) therapy has evolved rapidly over the past decade. Ongoing research has demonstrated a clear role for therapeutic GH in a wide spectrum of pediatric disorders involving both poor growth and abnormal body composition. Although guidelines for GH dosing are not fully established, a series of key studies has delineated the range of dosages that are useful in the treatment of children with growth disorders. The recent approval of idiopathic short stature (ISS) as an indication for GH therapy presents further challenges in optimizing the care of GH-treated patients. ISS is now recognized as a diverse collection of environmental and molecular abnormalities, some of which involve the GH-IGF axis. Emerging data indicate that serum IGF-I measurements are not only useful in the diagnosis of growth abnormalities but, in conjunction with auxological measurements, are also a powerful tool for assessing GH efficacy. While it is clear that many ISS patients respond to GH, some individuals will not show a satisfactory response. Monitoring IGF-I levels and change in height SDS during treatment can assist the physician in distinguishing those patients in whom GH successfully and safely induces statural growth from those with partial or complete GH insensitivity who might benefit from modified GH treatment protocols or alternate therapies. In addition, serum IGF-I measurements are increasingly used as part of a rational monitoring strategy to ensure safety of GH dosing in light of cumulative data associating high IGF-I levels with potential malignancy risk, and low IGF-I levels with cardiovascular disease risk. PMID- 16039894 TI - Proceedings of the 18th Annual National Cooperative Growth Study (NCGS) and the 5th Annual National Cooperative Somatropin Surveillance (NCSS) Investigator Meeting. October 14-16, 2004, Phoenix, Arizona, USA. PMID- 16039895 TI - Molecular characterization of Mycobacterium tuberculosis isolates in a region of Brazil with a high incidence of tuberculosis. AB - One hundred and seventy Mycobacterium tuberculosis clinical isolates were characterized by spoligotyping to evaluate the biodiversity of tubercle bacilli in a region of Brazil with a high incidence of tuberculosis (Pelotas and Rio Grande cities - Rio Grande do Sul State). The spoligotyping results were compared to the World Spoligotyping Database (Institut Pasteur de Guadeloupe), which contains data from >14,000 worldwide isolates of M. tuberculosis. The isolates clustered by spoligotyping were further characterized by IS6110-RFLP to confirm the clonal relationship. Sixty-six different spoligotypes were identified, grouping 125 of the isolates (74%). Approximately half of the isolates belonged to seven of the most frequently occurring spoligotypes in the database. Three shared types (with two or more isolates) not previously identified were given the type numbers 826, 827 and 863. An additional 45 spoligotypes were identified that did not match any existing database pattern. RFLP characterization reduced the number of isolates in most of the clusters, thereby showing a higher differentiation capacity than spoligotyping. These results highlight the importance of molecular epidemiology studies of tuberculosis in insufficiently studied regions with a high TB burden, in order to uncover the true extent of genetic diversity of the pathogen. PMID- 16039897 TI - Bacterial cell wall-expressed protein A triggers supraclonal B-cell responses upon in vivo infection with Staphylococcus aureus. AB - We have previously shown that staphylococcal protein A (SpA) anchored to the cell wall of Staphylococcus aureus acts as a virulence factor in septic arthritis. Apart from the ability of SpA to interact with Fcgamma, it also binds to Fab regions with immunoglobulin heavy chains encoded by the V(H) clan III gene family. The objective of the present study was to investigate whether in vivo expression of SpA by staphylococci induces V(H)III-dependent supraclonal B-cell responses, and whether such responses may affect the ability of the host to produce anti-staphylococcal antibodies. Upon primary infection of mice, a SpA expressing staphylococcal strain gave rise to significantly higher serum levels of V(H)III-encoded antibodies specific for SpA devoid of Fcgamma-binding ability (MSpA) than an isogeneic spa deletion mutant strain. The V(H)III-dependence of MSpA-specific antibody responses was affected by the size of the staphylococcal inoculum, and differed for IgM and IgG isotypes. Mice that had recovered from a prior mild infection from a SpA-expressing strain were protected against infection-induced weight loss upon reinfection. Although no lasting MSpA-specific IgG was induced by previous mild infection, these protected mice possessed IgG specific for clumping factor A, a conventional staphylococcal protein antigen. Our findings demonstrate that the expression of a B-cell superantigen during staphylococcal infection causes supraclonal changes to the immune system. Notably, while superantigen-triggered B-cell responses do not favor the development of SpA-specific memory B-cells, such responses do not interfere with the development of antibodies specific for a staphylococcal protein antigen associated with protective immunity. PMID- 16039898 TI - The varicella-zoster virus-mediated delayed host shutoff: open reading frame 17 has no major function, whereas immediate-early 63 protein represses heterologous gene expression. AB - We reported that varicella-zoster virus (VZV) causes a delayed host shutoff during its replicative cycle. VZV open reading frame 17 (ORF17) is the homologue of the herpes simplex virus (HSV) UL41 gene encoding the virion host shutoff (vhs) protein which is responsible for the shutoff effect observed in HSV infected cells. In the present study, we demonstrated that ORF17 is expressed as a late protein during the VZV replicative cycle in different infected permissive cell lines which showed a delayed shutoff of cellular RNA. A cell line with stable expression of VZV ORF17 was infected with VZV. In these cells, VZV replication and delayed host shutoff remained unchanged when compared to normal infected cells. ORF17 was not capable of repressing the expression of the beta gal reporter gene under the control of the human cytomegalovirus immediate-early gene promoter or to inhibit the expression of a CAT reporter gene under the control of the human GAPDH promoter, indicating that ORF17 has no major function in the VZV-mediated delayed host shutoff. To determine whether other viral factors are involved in the host shutoff, a series of cotransfection assays was performed. We found that the immediate-early 63 protein (IE63) was able to downregulate the expression of reporter genes under the control of the two heterologous promoters, indicating that this viral factor can be involved in the VZV-mediated delayed host shutoff. Other factors can be also implicated to modulate the repressing action of IE63 to achieve a precise balance between the viral and cellular gene expression. PMID- 16039899 TI - The Pseudomonas aeruginosa quorum-sensing molecule N-(3-oxododecanoyl)-L homoserine lactone stimulates phagocytic activity in human macrophages through the p38 MAPK pathway. AB - Quorum-sensing is an important mechanism for the regulation of bacteria-to bacteria communication. Recent advances have demonstrated that the Pseudomonas aeruginosa signaling molecule N-(3-oxododecanoyl)-L-homoserine lactone (3O-C(12) HSL) is also a potent modulator of eukaryotic cells and may thus play an important role in the host response during P. aeruginosa infections. Little is known, however, about specific effects of 3O-C(12)-HSL molecules on human macrophages. To address this issue, we investigated the influence of 3O-C(12)-HSL on the phagocytic activity, production of reactive oxygen species, and activation of p38 and p42/44 MAPK signaling pathways in human macrophages. We show an effect of 3O-C(12)-HSL on the phagocytic capacity in human macrophages, which depends on concentration and time of exposure. When cells were exposed to 100 microM 3O C(12)-HSL for 30 min or 1 h, the phagocytic activity increased 1.8 and 1.6 times, respectively. The 3O-C(12)-HSL treatments had no significant effect on the level of reactive oxygen species production. Furthermore, the p38 MAPK, but not the p42/44 MAPK, signaling pathway was activated in response to 3O-C(12)-HSL. In addition, specific blocking of p38 MAPK activation with 10 microM SB 203580 prevented the 3O-C(12)-HSL-induced increase in the phagocytic activity. These findings demonstrate that the bacterial quorum-sensing can play a significant role also in regulation of macrophage activity during infections caused by P. aeruginosa. PMID- 16039900 TI - Aquasonolysis of thioethers. AB - The aquasonolytical rate constants and products of diethyl sulfide, diallyl sulfide, dipropyl sulfide, dibutyl sulfide, diethyl disulfide, and dipropyl disulfide have been investigated. The sonolysis for model compounds in aqueous solutions follows the pseudo-first-order kinetics. Disulfides are more rapidly sonicated than sulfides. Neither vapour pressure nor thermostability of thioethers can obviously affect their aquasonolytical rates. Higher Henry's Law constant and hydrophobicity of substrate result in a higher aquasonolytical rate constant. Furthermore, the aquasonolytical rate constant decreases as the initial concentration of dipropyl sulfide increases. CS2, COS, thiols, thiirane, lower sulfides, butenyne, and butadiene are detected as principal very volatile products during ultrasonic irradiations. Thiophene, benzene, 1,5-hexadiene, and cyclopentadiene are additionally obtained during the aquasonolysis of diallyl sulfide. Furthermore, trisulfides, trithiolane, and other cyclic thiolanes as volatile products are observed. These results demonstrate that thioethers undergo thermal decomposition during their aquasonolyses. The transfer process of volatile thioethers between cavitation bubbles and bulk liquid remarkably affect the aquasonolysis, and the hydrophobicity of substrate dominates the transfer process. PMID- 16039901 TI - Molecular players regulating the jasmonate signalling network. AB - Many plant developmental and stress responses require the coordinated interaction of the jasmonate and other signalling pathways, such as those for ethylene, salicylic acid and abscisic acid. Recent research in Arabidopsis has uncovered several key players that regulate crosstalk between these signalling pathways and that shed light on the molecular mechanisms modulating this coordinated interaction. Genes that are involved in the regulation of protein stability through the ubiquitin-proteasome pathway (COI1, AXR1 and SGT1b), signalling proteins (MPK4) and transcription factors (AtMYC2, ERF1, NPR1 and WRKY70) form a regulatory network that allows the plant to fine-tune specific responses to different stimuli. PMID- 16039902 TI - QIAamp MinElute virus kit effectively extracts viral nucleic acids from cerebrospinal fluids and nasopharyngeal swabs. AB - BACKGROUND: Nucleic acid preparation from a variety of clinical specimens requires efficient target recovery and amplification inhibitor removal and is critical for successful molecular diagnosis. The QIAamp MinElute Virus kit (Qiagen Inc., Valencia, CA) was compared to the two existing methods currently used in our laboratory, IsoQuick (Orca Research Inc., Bothell, WA) for DNA extraction and RNAzol B (Leedo Laboratories Inc., Houston, TX) for RNA extraction, of viral nucleic acids. STUDY DESIGN: A total of 150 clinical specimens, including cerebrospinal fluid (CSF) and nasopharyngeal swabs (NPS), were used to determine the extraction efficiency of the MinElute compared to the other two methods. Nucleic acid recovery, hands-on time, turn-around-time and cost were compared across all kits. RESULTS: There was complete concordance between the MinElute and IsoQuick/RNAzol kits when herpes simplex virus (HSV), Epstein-Barr virus (EBV), varicella-zoster virus (VZV), influenza A virus or enteroviruses were detected using a colorimetric microtiter plate PCR system. The kits were equivalent in their ability to detect either DNA or RNA with superior ability to recover a high quality and quantity of RNA. With the potential to process larger specimen volumes, the MinElute kit can significantly shorten processing time from 2h to 50-55min. CONCLUSIONS: Although relatively high test kit costs were noted, the MinElute kit provides another rapid and user-friendly specimen processing tool in the diagnostic molecular microbiology laboratory. PMID- 16039903 TI - Lipid kinase activity of antibodies from milk of clinically healthy human mothers. AB - We have shown recently that polyclonal human milk sIgA contains a subfraction of antibodies (Abs) tightly bound to unusual minor milk lipids containing sialic acid. Here, we show that a small subfraction of milk IgG is tightly bound to the similar or the same minor lipids. The ability of small fractions of sIgA and IgG from human milk to phosphorylate selectively two minor lipids in the presence of [gamma-(32)P]nucleoside triphosphates was shown here for the first time to be an intrinsic property of these antibodies. In contrast to known kinases, antibodies with lipid kinase activity can transfer phosphoryl group to lipids not only from ATP but also from other different nucleotides (dATP, GTP, dGTP, UTP, TTP) with comparable efficiencies (30-100%). To our knowledge, there are no examples of enzymes using orthophosphate as a substrate of phosphorylation reactions. An extremely unusual property of lipid kinase Abs is their high affinity for orthophosphate (K(m)=1.6-5.6 microM) and capability to phosphorylate minor lipids using [(32)P]orthophosphate as donor of phosphate group. The relative specific activity and affinity of abzymes for orthophosphate and ATP depend significantly on donor milk. However, the levels of Ab-dependent phosphorylation of lipids for all Abs in the case of ATP (100%) and orthophosphate (60-80%) as substrates are comparable. The first example of natural abzymes with synthetic activity was milk sIgA with protein kinase activity. Most probably, lipid kinase sIgA and IgG of human milk are the second example of Abs with synthetic activity. PMID- 16039904 TI - The ins and outs of T-lymphocyte trafficking to the CNS: anatomical sites and molecular mechanisms. AB - This review addresses current knowledge of the molecular trafficking signals involved in the migration of circulating leukocytes across the highly specialized blood-central nervous system (CNS) barriers during immunosurveillance and inflammation. In this regard, adhesion molecules and activating and chemotactic factors are also discussed and the regional variability in the brain and spinal cord parenchyma are also considered. Furthermore, direct passage into cerebrospinal fluid (CSF) is discussed, in the context of CNS immunosurveillance. The potential differences that characterize leukocyte entry into these varied anatomical sites are highlighted, with special emphasis on studies of the pathogenesis of multiple sclerosis and its animal models. An update on findings from clinical trials of natalizumab is also provided. PMID- 16039905 TI - ProPred analysis and experimental evaluation of promiscuous T-cell epitopes of three major secreted antigens of Mycobacterium tuberculosis. AB - In the search for safe vaccine candidates against tuberculosis (TB), subunit vaccines including peptide-based candidates deserve consideration. However, an important requirement for such vaccine candidates is their promiscuous presentation to Th1 cells mediating protective immunity against TB, i.e. Th1 cells secreting IFN-gamma. The aim of the present study was to identify promiscuous Th1 cell epitopes of three major secreted antigens of Mycobacterium tuberculosis, i.e. ESAT-6, CFP10 and MPT70 by using a virtual matrix-based prediction program (ProPred) for peptide binding to 51 HLA-DR alleles. The ProPred analysis of these proteins was performed using the server (http:www.imtech.res.in/raghava/ProPed/). The peptides predicted to bind > 50% HLA-DR alleles included in the ProPred were considered promiscuous for binding predictions. Based on this criteria, one region in ESAT-6 (aa 69-77), two regions in CFP10 (aa 55-66 and aa 76-84) and four regions in MPT70 (aa 1-11, aa 81-95, aa 124-140 and aa 182-191) were considered promiscuous HLA-DR binders. The experimental evaluation of these regions, by using overlapping synthetic peptides for presentation to T-cells, confirmed the promiscuous nature of peptides covering the regions aa 69-77, aa 76-84 and aa 182-191 of ESAT-6, CFP10 and MPT70, respectively. These results demonstrate that the ProPred analysis can facilitate the selection of promiscuous peptides recognized by Th1 cells, and thus it can be useful in the identification of peptide-based vaccine candidates against TB. PMID- 16039906 TI - Exploiting scale-free information from expression data for cancer classification. AB - Most studies concerning expression data analyses usually exploit information on the variability of gene intensity across samples. This information is sensitive to initial data processing, which affects the final conclusions. However expression data contains scale-free information, which is directly comparable between different samples. We propose to use the pairwise ratio of gene expression values rather than their absolute intensities for a classification of expression data. This information is stable to data processing and thus more attractive for classification analyses. In proposed schema of data analyses only information on relative gene expression levels in each sample is exploited. Testing on publicly available datasets leads to superior classification results. PMID- 16039907 TI - Custom zinc-finger nucleases for use in human cells. AB - Genome engineering through homologous recombination (HR) is a powerful instrument for studying biological pathways or creating treatment options for genetic disorders. In mammalian cells HR is rare but the creation of targeted DNA double strand breaks stimulates HR significantly. Here, we present a method to generate, evaluate, and optimize rationally designed endonucleases that promote HR. The DNA binding domains were synthesized by assembling predefined zinc-finger modules selected by phage display. Attachment of a transcriptional activation domain allowed assessment of DNA binding in reporter assays, while fusion with an endonuclease domain created custom nucleases that were tested for their ability to stimulate HR in episomal and chromosomal gene repair assays. We demonstrate that specificity, expression kinetics, and protein design are crucial parameters for efficient gene repair and that our two-step assay allows one to go quickly from design to testing to successful employment of the custom nucleases in human cells. PMID- 16039908 TI - Glucagon-like peptide-1 plasmid construction and delivery for the treatment of type 2 diabetes. AB - Glucagon-like peptide-1 (GLP-1) is a 30-amino-acid hormone produced by intestinal L cells. It has been proposed that GLP-1 can be used as a new treatment for type 2 diabetes mellitus because it acts to augment insulin secretion and its effectiveness is maintained in type 2 diabetic patients. Despite its many remarkable advantages as a therapeutic agent for diabetes, GLP-1 is not immediately clinically applicable because of its extremely short half-life. One way to overcome this drawback is GLP1 gene delivery, which enables GLP-1 production in the body. In this study, the effect of GLP1 gene delivery was evaluated both in vitro and in vivo using a new plasmid constructed with a GLP1 (7-37) cDNA. The expression of the GLP1 gene was driven by a SV40 promoter/enhancer. To increase the expression level of GLP-1, nuclear factor kappaB binding sites were introduced. The in vitro results showed expression of GLP-1 and in vitro activity of GLP-1, which is a glucose-dependent insulinotropic action. A single systemic administration of polyethyleneimine/pSIGLP1NFkappaB complex into DIO mice resulted in increasing insulin secretion and decreasing blood glucose levels for a duration longer than 2 weeks. PMID- 16039909 TI - T Lymphocytes transduced with a lentiviral vector expressing F12-Vif are protected from HIV-1 infection in an APOBEC3G-independent manner. AB - The viral infectivity factor (Vif) is an essential component of the HIV-1 infectious cycle. Vif counteracts the action of the cytidine deaminase APOBEC3G (AP3G), which confers nonimmune antiviral defense against HIV-1 to T lymphocytes. Disabling or interfering with the function of Vif could represent an alternative therapeutic approach to AIDS. We have expressed a natural mutant of Vif, F12-Vif, in a VSV-G-pseudotyped lentiviral vector under the Tat-inducible control of the HIV-1 LTR. Conditional expression of F12-Vif prevents replication and spreading of both CXCR4 and CCR5 strains of HIV-1 in human primary T lymphocyte and T cell lines. T cells transduced with F12-Vif release few HIV-1 virions and with reduced infectivity. Several lines of evidence indicate that HIV-1 interference requires the presence of both wild-type and F12-Vif proteins, suggesting a dominant negative feature of the F12-Vif mutant. Surprisingly, however, the F12-Vif mediated inhibition does not depend on the reestablishment of the AP3G function. PMID- 16039910 TI - Gender and age related differences in left ventricular function and geometry with focus on the long axis. AB - AIMS: To study age and gender related alterations in left ventricular (LV) long axis function. METHODS: Eighty-two healthy individuals from the general population in three age groups were investigated. LV long axis and short axis function and dimensions were studied with echocardiographic M-mode and two dimensional technique. RESULTS: The most prominent age related differences were observed in LV long axis function, whereas only minor alterations in short axis function were noticed. Both systolic and diastolic long axis function decreased with advancing age; maximal systolic velocity (r=0.61, p<0.0001), maximal early diastolic filling velocity (r=0.87, p<0.0001). The length of the long axis decreased with age, while the relative contraction amplitude was maintained. LV global and short axis measurements revealed significant differences between genders, males having generally larger dimensions, even when correcting for body surface area. Females exhibited a more pronounced remodelling process with advancing age. CONCLUSION: Functional age related changes in LV function are more prominent in the long axis, while differences between genders are more pronounced in short axis and in volume measurements. These findings might be of importance when remodelling processes are evaluated, as these appear to be different in men and women and also age related. PMID- 16039911 TI - Micro pulsed radio-frequency electroporation chips. AB - Electroporation (EP) is one of the most important physical methods in biotechnology, which employs electrical pulses to transiently permeabilize cell membranes. In this study, a new micro pulsed radio-frequency electroporation cell (microPREP) chip was fabricated using a lift-off technique and SU-8 photolithography. The biological tests were carried out using three different plant protoplasts (cabbage, spinach and oil rape) on the micro EP chip and a pulsed RF electric field was applied to the microchip. The variations of fluorescent intensity and cell viability as functions of the electric pulse amplitude and duration time during the electroporation process were studied in detail at the single-cell level. Using such chip design and test method, one can easily optimize the efficiency and cell viability. Also, a large amount of statistical data can be quickly obtained. Finally, results of this parametric study were presented in the "phase diagram", from which the critical electric field for inducing single-cell electroporation under different conditions can be clearly determined. PMID- 16039912 TI - Synthetic oligonucleotides: AFM characterisation and electroanalytical studies. AB - One of the most important steps in designing more sensitive and stable DNA based biosensors is the immobilisation procedure of the nucleic acid probes on the transducer surface, while maintaining their conformational flexibility. MAC Mode AFM images in air demonstrated that the oligonucleotide sequences adsorb spontaneously on the electrode surface, showing the existence of pores in the adsorbed layer that reveal big parts of the electrode surface, which enables non specific adsorption of other molecules on the uncovered areas. The electrostatic immobilisation onto a glassy carbon electrode followed by hybridisation with a complementary sequence and control with a non-complementary sequence was studied using differential pulse voltammetry and electrochemical impedance spectroscopy. Changes in the oxidation currents of guanosine and adenosine were observed after hybridisation events as well as after control experiments. Modification of the double layer capacitance that took place after hybridisation or control experiments showed that non-specific adsorption of complementary or non complementary sequences occur allowing the formation of a mixed multilayer. PMID- 16039913 TI - Intergenerational transfers may have decoupled physiological and chronological age in a eusocial insect. AB - Life-history theory generally predicts that there should be no selection for longevity beyond the limit of reproductive capacity. However, the capacity to increase fitness may not end when individuals reach a state of functional sterility. Recent studies show that intergenerational transfers of resources from post-reproductive parents can increase the offspring's fitness, and analytical theory shows that age-trajectories of transfers may shape the course of senescence in social organisms. In eusocial insects, female roles are partitioned so that one phenotype or "caste" reproduces while another is responsible for resource transfers: the reproductive "queens" are arrested in a continuous reproductive mode, while transfer-activities such as hygienic behaviors, guarding, foraging and further food processing ("nursing") that increases the nutritional value of provisions are conducted by sterile "workers". Worker honey bees normally perform these tasks in a sequence so that nursing inside the protected nest is conducted prior to more risky exterior hive activities such as guarding and foraging. However, foragers may revert to nurse-activity in response to demographic changes, and worker bees can also develop into a stress resistant survival form with a 10-fold increase in lifespan. This elastic division of parental functions is believed to increase colony fitness. Further, it generates a stage-dependent trajectory of senescence that is difficult to address with established theories of aging. In the following, we show how a recent theory that includes resource transfers can be used to elucidate patterns of senescence in eusocial, non-reproducing individuals like the honey bee worker. PMID- 16039914 TI - Saccharomyces cerevisiae RAD53 (CHK2) but not CHK1 is required for double-strand break-initiated SCE and DNA damage-associated SCE after exposure to X rays and chemical agents. AB - Saccharomyces cerevisiae RAD53 (CHK2) and CHK1 control two parallel branches of the RAD9-mediated pathway for DNA damage-induced G(2) arrest. Previous studies indicate that RAD9 is required for X-ray-associated sister chromatid exchange (SCE), suppresses homology-directed translocations, and is involved in pathways for double-strand break repair (DSB) repair that are different than those controlled by PDS1. We measured DNA damage-associated SCE in strains containing two tandem fragments of his3, his3-Delta5' and his3-Delta3'::HOcs, and rates of spontaneous translocations in diploids containing GAL1::his3-Delta5' and trp1::his3-Delta3'::HOcs. DNA damage-associated SCE was measured after log phase cells were exposed to methyl methanesulfonate (MMS), 4-nitroquinoline 1-oxide (4 NQO), UV, X rays and HO-induced DSBs. We observed that rad53 mutants were defective in MMS-, 4-NQO, X-ray-associated and HO-induced SCE but not in UV associated SCE. Similar to rad9 pds1 double mutants, rad53 pds1 double mutants exhibited more X-ray sensitivity than the single mutants. rad53 sml1 diploid mutants exhibited a 10-fold higher rate of spontaneous translocations compared to the sml1 diploid mutants. chk1 mutants were not deficient in DNA damage associated SCE after exposure to DNA damaging agents or after DSBs were generated at trp1::his3-Delta5'his3-Delta3'::HOcs. These data indicate that RAD53, not CHK1, is required for DSB-initiated SCE, and DNA damage-associated SCE after exposure to X-ray-mimetic and UV-mimetic chemicals. PMID- 16039916 TI - Small bowel transplantation for diffuse intestinal angiomatosis. PMID- 16039917 TI - Radiological digital teaching file development: an overview. AB - Radiologists are collectors of interesting films for teaching purposes or for use in presentations and publications. Traditionally, hard copies of films have been stored in an organized fashion, usually in a filing cabinet or film library. This system has inherent limitations, such as the physical space required. Many of the shortcomings can be circumvented by development of an electronic teaching file. Whereas the implementation of an institutional radiological digital image database can require significant developmental effort and programming expertise, there are a number of web-based solutions which are freely available and can be relatively easily employed to establish a contemporary electronic image library. This article will review the various options and discuss the process of developing a digital image database. PMID- 16039918 TI - Neuroradiology. AB - This is the second in a series of short reviews of internet-based radiological educational resources, and will focus on neuroradiology. Most of the sites cater for medical students and trainee or non-specialist radiologists, but may also be of interest to specialists, especially for use in teaching. Hyperlinks are available in the electronic version of this article and were all active at the time of going to press (August 2005). If your computer has high-level internet security settings or does not accept cookies, you may not be able to access all of the sites listed; this may be the case on some hospital-based systems. All websites should be fully accessible from other computers with less stringent security settings, e.g. a PC at home. PMID- 16039919 TI - Teleradiology: threat or opportunity? AB - The rapid advances in information technology and communication bandwidth have spawned an equally rapid development of clinical teleradiology. Current computer technology and communication capability allow easy transfer of diagnostic images, of any complexity, to any location in the world. This provides the opportunity to acquire swift primary and secondary diagnostic opinions from the remotest of locations, often at economically attractive rates, with the potential for easing the burden on hard-pressed departments of radiology. However, this comes at the potential cost of distancing the clinical radiologist from the patient, with consequent impact upon direct clinical care. As this technology advances across the world, it is vital that UK radiologists are familiar with the clinical implications, the medicolegal framework within which the field operates and the associated governance issues. This paper reviews current practice and discusses the associated risks. PMID- 16039920 TI - Radiological appearances in the pelvis following rectal cancer surgery. AB - Radiology has a significant role in the evaluation of surgery for rectal cancer. With recent developments in surgical techniques, the number of neorectal reservoir configurations has increased. It is important to recognize the normal and abnormal appearances, both early and late, following pelvic surgery. The aim of this pictorial review is to demonstrate the imaging techniques that are used in both the investigation and the follow-up of patients who have undergone uncomplicated or complicated rectal resection. PMID- 16039921 TI - Vascular masses of the head and neck. AB - The classification, clinical presentation, natural history and imaging appearances of vascular masses of the head and neck are reviewed. The radiological appearances of congenital vascular masses (infantile haemangiomas and other congenital vascular tumours, high-flow and low-flow vascular malformations), acquired vascular masses (benign and malignant tumours, non neoplastic lesions) and other hypervascular masses are described and illustrated, together with consideration of image-guided interventions. PMID- 16039922 TI - Cross-sectional imaging in non-melanoma skin cancer of the head and neck. AB - AIM: To investigate in head and neck non-melanoma skin cancers (NMSCs) the accuracy of cross-sectional imaging for detection of local tumour extent, recurrent tumour and prediction of patient outcome. METHODS: This retrospective study included 33 NMSC patients (22 men, 11 women, median age 69 years) with 8 primary and 25 suspected recurrent tumours. The findings of magnetic resonance imaging (MRI) and computed tomography (CT) were compared with histopathology, and accuracy of MRI or CT in detecting local recurrence was determined. Extent of disease on imaging was compared with patient outcome assessed by clinical follow up to a mean of 26.4 months. RESULTS: Lesions were identified in 29 patients, whose mean disease-free survival (DFS) was 25.5 months. In 4 of these cases, where imaging showed no invasion of deep structures, DFS was 56 months. In the other 25 cases DFS was 20.6 months, irrespective of treatment but varying with site of involvement. Of 19 patients treated with surgery, imaging of 16 showed deep invasion, which was confirmed at histology in 15 (93.7% accuracy), and 3 had superficial tumours on imaging all confirmed by histology (100% accuracy). Imaging accuracy for identifying recurrent tumour was 96% (24 of 25 patients). CONCLUSION: In NMSC, cross-sectional imaging accurately identifies tumour extent and local recurrence. The extent of disease and invasion of deeper structures predicts patient outcome. PMID- 16039923 TI - The contribution of high-resolution multiplanar reformats of the skull base to the detection of skull-base fractures. AB - AIM: To investigate the contribution of routine review of submillimetric multiplanar reformats to the diagnosis of skull-base fractures. METHODS: A prospective analysis was performed of 407 cases referred over a 6-month period for CT of the skull following cranial trauma. The reformatted 5-mm axial sections and subsequently the high-resolution multiplanar reformats (HRMPRs) were viewed on an ADW 4.1 workstation using bone windows and algorithm. All skull-base fractures and related features, recorded by the consensus of two radiologists, were classified as anatomically significant or non-significant on the basis of eight criteria. The clinical features of skull-base injury and any subsequent treatment were noted in all cases of skull-base fracture. RESULTS: HRMPRs detected 80 separate skull-base fractures in 36/407 cases. Of these 80 fractures, 57 were visible on 5-mm axial sections. In 8 of the 36 cases, the significant anatomical features were only evident on review of the HRMPRs. In 6 of the 36 cases, none of the skull-base fractures was visible on 5-mm sections, but these individuals had only minor associated clinical features and no therapeutic requirements. Review of HRMPRs could have been confined to patients with skull base fractures, abnormal intracranial and extracranial air collections or opacified mastoid air cells revealed by 5-mm axial sections. This policy would have led to the detection of 79/80 (99%) of skull-base fractures and all significant anatomical features. CONCLUSION: The 5-mm axial sections demonstrated 71% of skull-base fractures and 78% of skull-base fractures with significant anatomical features, using HRMPRs as a gold standard. There were no significant clinical sequelae at short-term follow-up of those fractures only evident on HRMPRs. PMID- 16039924 TI - Imaging features of retroperitoneal and pelvic schwannomas. AB - AIM: To describe the imaging features of retroperitoneal and pelvic schwannomas. MATERIALS AND METHODS: The presenting cross-sectional imaging for 18 sequential patients with retroperitoneal or pelvic schwannomas was reviewed retrospectively. Note was made of tumour diameter, position, homogeneity, margin, shape, calcification and invasion into adjacent structures. Where MRI had been performed, T1 and T2 signal intensity relative to skeletal muscle, and the degree and pattern of enhancement with gadolinium, were also assessed. RESULTS: Imaging from 13 patients was available for review. The mean tumour diameter was 8.7 cm (range 4 to 15 cm); 9 schwannomas were located in the pelvis and 4 in the retroperitoneum; 12 cases showed smooth, regular margins and 1 case irregular, invasive margins. The tumours were homogeneous in 5 cases and heterogeneous with cystic change in 8; in 2 cases there was smooth expansion of a sacral nerve root exit foramen, and in 1 there was bony destruction of the sacrum and extension of tumour into the spinal canal. In 5 cases MRI was performed; on T1-weighted images all tumours were isointense; on T2-weighted images 4 tumours were hyperintense and 1 was isointense to skeletal muscle. In all cases the diagnosis was confirmed by core biopsy. CONCLUSION: Retroperitoneal and pelvic schwannomas typically form large, well-circumscribed masses in the retroperitoneum or presacral area, and frequently undergo cystic degeneration. They can occasionally cause bony changes in the spine, but otherwise do not invade or obstruct adjacent structures. Although they are rare, it is important for the radiologist to recognize the typical appearance of schwannomas because they can be mistaken for malignant tumours. PMID- 16039925 TI - Orbital complications of functional endoscopic sinus surgery: MR and CT findings. AB - AIM: To describe the radiological findings of ophthalmic complications during functional endoscopic sinus surgery (FESS) and correlate them with the clinical manifestations and mechanisms of injury. METHODS: This was a retrospective review of the clinical and cross-sectional imaging findings of 9 patients with orbital complications during FESS. RESULTS: The most common site of entry into the orbit during FESS was the lower medial orbital wall (7 of 9), followed by the inferior orbital wall, resulting in injury to the medial rectus (4 of 9) and, less frequently, the inferior rectus (2 of 9) or superior oblique muscles (1 of 9). Extensive scarring on imaging (3 of 9) was associated with global ocular motility dysfunction. In contrast, localized scarring (3 of 9) or extraocular muscle trauma (6 of 9) resulted in disturbance of eye movement in the direction of gaze from the injured site. CONCLUSIONS: Orbital magnetic resonance and computed tomography findings correlate very well with the abnormal eye movements clinically observed, and can assist in clarifying the cause of injury and guide surgical corrective management of patients suffering orbital complications from FESS. Radiologists should be familiar with the recent developments in FESS instrumentation as well as with the most commonly injured structures within the orbit. PMID- 16039926 TI - Accuracy and feasibility of dynamic contrast-enhanced 3D MR imaging in the assessment of lung perfusion: comparison with Tc-99 MAA perfusion scintigraphy. AB - AIM: The aim of this study was to correlate findings of perfusion magnetic resonance imaging (MRI) and perfusion scintigraphy in cases where there was a suspicion of abnormal pulmonary vasculature, and to evaluate the usefulness of MRI in the detection of perfusion deficits of the lung. METHODS: In all, 17 patients with suspected abnormality of the pulmonary vasculature underwent dynamic contrast-enhanced MRI. T1-weighted 3D fast-field echo pulse sequences were obtained (TR/TE 3.3/1.58 ms; flip angle 30 degrees; slice thickness 12 to 15 mm). The dynamic study was acquired in the coronal plane following administration of 0.1 mmol/kg gadopentetate dimeglumine. A total of 8 to 10 sections repeated 20 to 25 times at intervals of 1s were performed. Perfusion lung scintigraphy was carried out a maximum of 48 h before the MR examination in all cases. Two radiologists, who were blinded to the clinical data and results of other imaging methods, reviewed all coronal sections. MR perfusion images were independently assessed in terms of segmental or lobar perfusion defects in the 85 lobes of the 17 individuals, and the findings were compared with the results of scintigraphy. RESULTS: Of the 17 patients, 8 were found to have pulmonary emboli, 2 chronic obstructive pulmonary disease with emphysema, 2 bullous emphysema, 2 Takayasu arteritis and 1 had a hypoplastic pulmonary artery. Pulmonary perfusion was completely normal in 2 cases. In 35 lobes, perfusion defects were detected using both methods, in 4 with MR alone and in 9 only with scintigraphy. There was good agreement between MRI and scintigraphy findings (kappa=0.695). CONCLUSION: Pulmonary perfusion MRI is a new alternative to scintigraphy in the evaluation of pulmonary perfusion for various lung disorders. In addition, this technique allows measurement and quantification of pulmonary perfusion abnormalities. PMID- 16039927 TI - The use of radiological guidelines to achieve a sustained reduction in the number of radiographic examinations of the cervical spine, lumbar spine and knees performed for GPs. AB - AIM: To determine if the use of request guidelines can achieve a sustained reduction in the number of radiographic examinations of the cervical spine, lumbar spine and knee joints performed for general practitioners (GPs). METHODS: GPs referring to three community hospitals and a district general hospital were circulated with referral guidelines for radiography of the cervical spine, lumbar spine and knee, and all requests for these three examinations were checked. Requests that did not fit the guidelines were returned to the GP with an explanatory letter and a further copy of the guidelines. Where applicable, a large-joint replacement algorithm was also enclosed. If the GP maintained the opinion that the examination was indicated, she or he had the option of supplying further justifying information in writing or speaking to a consultant radiologist. RESULTS: Overall the number of radiographic examinations fell by 68% in the first year, achieving a 79% reduction in the second year. For knees, lumbar spine and cervical spine radiographs the total reductions were 77%, 78% and 86%, respectively. CONCLUSION: The use of referral guidelines, reinforced by request checking and clinical management algorithms, can produce a dramatic and sustained reduction in the number of radiographs of the cervical spine, lumbar spine and knees performed for GPs. PMID- 16039928 TI - Diagnostic value of bronchoscopy, CT and transbronchial biopsies in diffuse pulmonary lymphangiomatosis: case report and review of the literature. AB - The authors present the case of a 48-year-old man with diffuse pulmonary lymphangiomatosis. This rare lymphatic disorder is characterized by proliferation of anastomosing lymphatic vessels varying in size. Clinical presentation and imaging findings are highly suggestive. Bronchoscopic examination of this patient showed, for the first time to our knowledge, vesicles disseminated throughout the bronchial tree. Histopathological examinations are necessary to differentiate lymphangiomatosis from lymphangiectasis. The diagnosis can be made by transbronchial biopsy without performing open lung biopsy which was, until now, considered necessary for diagnosis. PMID- 16039929 TI - Percutaneous coaxial needle biopsy using the spacer technique: a novel and easy modification to increase procedure safety. AB - We describe a novel but easy modification to percutaneous coaxial needle biopsy technique using readily available standard equipment to enable a predefined variable sampling length to be achieved. This technique involves the insertion of a carefully measured spacer between the coaxial cutting needle and guide needle. This can minimise the sampling length required to biopsy any given lesion, minimising the volume of traumatised tissue and preventing unnecessary penetration of tissues deep to smaller lesions, thus increasing procedure safety. PMID- 16039930 TI - Complex injuries from a gunshot injury to the upper abdomen. Have we moved to the post surgery era? PMID- 16039931 TI - The value of ultrasonographic examination of the lumbar spine in infants with specific reference to cutaneous markers of occult spinal dysraphism. PMID- 16039932 TI - A cross-site vascular radiology on-call service: the Manchester experience. PMID- 16039933 TI - X-ray dose training: are we exposed to enough? A reply to Dr McCoubrie's letter published in Vol 60 No 6 (2005) on p. 730. PMID- 16039934 TI - Eyetracking and selective attention in category learning. AB - An eyetracking version of the classic Shepard, Hovland, and Jenkins (1961) experiment was conducted. Forty years of research has assumed that category learning often involves learning to selectively attend to only those stimulus dimensions useful for classification. We confirmed that participants learned to allocate their attention optimally. We also found that learners tend to fixate all stimulus dimensions early in learning. This result obtained despite evidence that participants were also testing one-dimensional rules during this period. Finally, the restriction of eye movements to only relevant dimensions tended to occur only after errors were largely (or completely) eliminated. We interpret these findings as consistent with multiple-systems theories of learning which maximize information input in order to maximize the number of learning modules involved, and which focus solely on relevant information only after one module has solved the learning problem. PMID- 16039936 TI - Health and the hot air of a European summer. PMID- 16039935 TI - On the capacity of attention: its estimation and its role in working memory and cognitive aptitudes. AB - Working memory (WM) is the set of mental processes holding limited information in a temporarily accessible state in service of cognition. We provide a theoretical framework to understand the relation between WM and aptitude measures. The WM measures that have yielded high correlations with aptitudes include separate storage-and-processing task components, on the assumption that WM involves both storage and processing. We argue that the critical aspect of successful WM measures is that rehearsal and grouping processes are prevented, allowing a clearer estimate of how many separate chunks of information the focus of attention circumscribes at once. Storage-and-processing tasks correlate with aptitudes, according to this view, largely because the processing task prevents rehearsal and grouping of items to be recalled. In a developmental study, we document that several scope-of-attention measures that do not include a separate processing component, but nevertheless prevent efficient rehearsal or grouping, also correlate well with aptitudes and with storage-and-processing measures. So does digit span in children too young to rehearse. PMID- 16039937 TI - Factors influencing GPs' decisions regarding screening for high alcohol consumption: a focus group study in Swedish primary care. AB - OBJECTIVE: The aim of this study was to explore factors that influence general practitioners' (GPs') decisions regarding screening for high alcohol consumption. METHODS: GPs working at three primary healthcare centres in Sweden participated in focus group interviews. The interviews were recorded and transcribed verbatim, and a deductive framework approach was used for the analysis. RESULTS: The majority of the participating GPs did not believe in asking all patients about their alcohol consumption. Reported factors that influenced how many and which patients were questioned about alcohol consumption were time, age of the patient, consultation setting, patient-physician relationship, what symptoms the patient presented with, and knowledge of measures if patients appear to have a high alcohol consumption. Thus, alcohol screening and intervention were not performed in all patient groups as was originally intended, but were performed in limited groups of patients such as those with alcohol-related symptoms. CONCLUSIONS: Although the number of participants in this study was small and the conclusions cannot be generalized, the results provide some valuable insights into why GPs are hesitant to engage in screening for high alcohol consumption. Since prevention of alcohol-related health problems is an important public health issue, many different screening and intervention strategies have to be formulated and evaluated in order to reach patients with both hazardous and harmful alcohol consumption within the healthcare system. Screening all consecutive patients for a limited period or screening patient groups known to include a fairly high frequency of high alcohol consumers are two ways of limiting the time requirements and increasing role legitimacy. Still, there is a need for a broader public health strategy involving many players in the community in alcohol preventive measures, especially in more primary preventive approaches. PMID- 16039938 TI - Ozone air pollution and daily mortality in Genoa, Italy between 1993 and 1996. AB - The association between ozone (O3) and daily mortality was investigated in Genoa, an Italian city characterized by a Mediterranean climate and a high prevalence of elderly inhabitants. The O3 effect, adjusted for long time trend, seasonality and weather, was assessed using Poisson regression modelling, allowing for overdispersion and autocorrelation, and expressed as mean variation percent of daily mortality per 50 microg/m3 increase (MV). Significant MVs for overall (+4.0%) and cardiovascular (+7.2%) mortality were detected at 1-day lag. The effects were stronger in the warmer season (May-October). Similar estimates were found after restricting the analyses to the elderly (>or=75 years). Furthermore, in this group, higher MVs were observed for total mortality at 2-day lag. A statistically significant synergistic effect between O3 and temperature was observed for cardiovascular mortality, particularly in elderly people, with an evident increase in mortality risk above 26 degrees C (MV +30.0% for the whole population and +40.0% for the elderly, respectively). This investigation highlights the importance of taking local climatic and demographic features into account when comparing different time-series studies, and substantiates the influence of photochemical pollution on mortality trends in small urban areas. PMID- 16039939 TI - Nickel decreases cellular iron level and converts cytosolic aconitase to iron regulatory protein 1 in A549 cells. AB - Nickel (Ni) compounds are well-established carcinogens and are known to initiate a hypoxic response in cells via the stabilization and transactivation of hypoxia inducible factor-1 alpha (HIF-1alpha). This change may be the consequence of nickel's interference with the function of several Fe(II)-dependent enzymes. In this study, the effects of soluble nickel exposure on cellular iron homeostasis were investigated. Nickel treatment decreased both mitochondrial and cytosolic aconitase (c-aconitase) activity in A549 cells. Cytosolic aconitase was converted to iron-regulatory protein 1, a form critical for the regulation of cellular iron homeostasis. The increased activity of iron-regulatory protein 1 after nickel exposure stabilized and increased transferrin receptor (Tfr) mRNA and antagonized the iron-induced ferritin light chain protein synthesis. The decrease of aconitase activity after nickel treatment reflected neither direct interference with aconitase function nor obstruction of [4Fe-4S] cluster reconstitution by nickel. Exposure of A549 cells to soluble nickel decreased total cellular iron by about 40%, a decrease that likely caused the observed decrease in aconitase activity and the increase of iron-regulatory protein 1 activity. Iron treatment reversed the effect of nickel on cytosolic aconitase and iron-regulatory protein 1. To assess the mechanism for the observed effects, human embryonic kidney (HEK) cells over expressing divalent metal transporter-1 (DMT1) were compared to A549 cells expressing only endogenous transporters for inhibition of iron uptake by nickel. The inhibition data suggest that nickel can enter via DMT1 and compete with iron for entry into the cell. This disturbance of cellular iron homeostasis by nickel may have a great impact on the ability of the cell to regulate a variety of cell functions, as well as create a state of hypoxia in cells under normal oxygen tension. These effects may be very important in how nickel exerts phenotypic selection pressure to convert a normal initiated cell into a cancer cell. PMID- 16039940 TI - Molecular events associated with arsenic-induced malignant transformation of human prostatic epithelial cells: aberrant genomic DNA methylation and K-ras oncogene activation. AB - Numerous studies link arsenic exposure to human cancers in a variety of tissues, including the prostate. Our prior work showed that chronic arsenic exposure of the non-tumorigenic, human prostate epithelial cell line, RWPE-1, to low levels of (5 microM) sodium arsenite for 29 weeks resulted in malignant transformation and produced the tumorigenic CAsE-PE cell line. The present work focuses on the molecular events occurring during this arsenic-induced malignant transformation. Genomic DNA methylation was significantly reduced in CAsE-PE cells. A time course experiment showed that during malignant transformation DNA methyltransferase activity was markedly reduced by arsenic. However, DNA methyltransferase mRNA levels were not affected by arsenic exposure. Microarray screening showed that K ras was highly overexpressed in CAsE-PE cells, a result further confirmed by Northern blot and Western blot analyses. Since ras activation is thought to be a critical event in prostate cancer progression, further detailed study was performed. Time course experiments also showed that increased K-ras expression preceded malignant transformation. Mutational analysis of codons 12, 13, and 61 indicated the absence of K-ras mutations. The K-ras gene can be activated by hypomethylation, but our study showed that CpG methylation in K-ras promoter region was not altered by arsenic exposure. Arsenic metabolism studies showed RWPE-1, CAsE-PE, and primary human prostate cells all had a very poor capacity for arsenic methylation. Thus, inorganic arsenic-induced transformation in human cells is associated with genomic DNA hypomethylation and K-ras overexpression. However, overexpression of K-ras occurred without mutations and through a mechanism other than promoter region hypomethylation. PMID- 16039941 TI - Arsenic exposure, urinary arsenic speciation, and peripheral vascular disease in blackfoot disease-hyperendemic villages in Taiwan. AB - Long-term exposure to ingested inorganic arsenic is associated with peripheral vascular disease (PVD) in the blackfoot disease (BFD)-hyperendemic area in Taiwan. This study further examined the interaction between arsenic exposure and urinary arsenic speciation on the risk of PVD. A total of 479 (220 men and 259 women) adults residing in the BFD-hyperendemic area were studied. Doppler ultrasound was used to diagnose PVD. Arsenic exposure was estimated by an index of cumulative arsenic exposure (CAE). Urinary levels of total arsenic, inorganic arsenite (As(III)) and arsenate (As(V)), monomethylarsonic acid (MMA(V)), and dimethylarsinic acid (DMA(V)) were determined. Primary methylation index [PMI = MMA(V)/(As(III) + As(V))] and secondary methylation index (SMI = DMA(V)/MMA(V)) were calculated. The association between PVD and urinary arsenic parameters was evaluated with consideration of the interaction with CAE and the confounding effects of age, sex, body mass index, total cholesterol, triglycerides, cigarette smoking, and alcohol consumption. Results showed that aging was associated with a diminishing capacity to methylate inorganic arsenic and women possessed a more efficient arsenic methylation capacity than men did. PVD risk increased with a higher CAE and a lower capacity to methylate arsenic to DMA(V). The multivariate adjusted odds ratios for CAE of 0, 0.1-15.4, and >15.4 mg/L x year were 1.00, 3.41 (0.74-15.78), and 4.62 (0.96-22.21), respectively (P < 0.05, trend test); and for PMI < or = 1.77 and SMI > 6.93, PMI > 1.77 and SMI > 6.93, PMI > 1.77 and SMI < or = 6.93, and PMI < or = 1.77 and SMI < or = 6.93 were 1.00, 2.93 (0.90 9.52), 2.85 (1.05-7.73), and 3.60 (1.12-11.56), respectively (P < 0.05, trend test). It was concluded that individuals with a higher arsenic exposure and a lower capacity to methylate inorganic arsenic to DMA(V) have a higher risk of developing PVD in the BFD-hyperendemic area in Taiwan. PMID- 16039942 TI - A mathematical model for the absorption and metabolism of formaldehyde vapour by humans. AB - Epidemiological studies of occupational exposure to formaldehyde gas (HCHO) have suggested possible links between concentration and duration of exposure, and elevated risks of leukaemia and other cancers at sites distant from the site of contact. Formaldehyde is a highly water soluble gas which, when inhaled, reacts rapidly at the site of contact and is quickly metabolised by enzymes in the respiratory tissue. Inhaled formaldehyde is almost entirely absorbed in the respiratory tract and, for formaldehyde induced toxicity to occur at distant sites, HCHO must enter the blood and be transported to systemic tissues via the circulatory system. A mathematical model describing the absorption and removal of inhaled formaldehyde in the nasal tissue is therefore formulated to predict the proportion of formaldehyde entering into the blood. Accounting for the spatial distribution of the formaldehyde concentration and the metabolic activity within the mucosa, the concentration of formaldehyde in the mucus, the epithelium and the blood has been determined and was found to attain a steady-state profile within a few seconds of exposure. The increase of the formaldehyde concentration in the blood was predicted to be insignificant compared with the existing pre exposure levels in the body, indicating that formaldehyde is rapidly removed in the nasal tissue. The results of the model thus suggest that it is highly unlikely that following inhalation by the nose, formaldehyde itself will cause toxicity at sites other than the initial site of contact in the respiratory tract. PMID- 16039943 TI - Gentamicin-induced apoptosis in LLC-PK1 cells: involvement of lysosomes and mitochondria. AB - Gentamicin accumulates in lysosomes and induces apoptosis in kidney proximal tubules and renal cell lines. Using LLC-PK1 cells, we have examined the concentration- and time-dependency of the effects exerted by gentamicin (1-3 mM; 0-3 days) on (i) lysosomal stability; (ii) activation of mitochondrial pathway; (iii) occurrence of apoptosis (concentrations larger than 3 mM caused extensive necrosis as assessed by the measurement of lactate dehydrogenase release). Within 2 h, gentamicin induced a partial relocalization [from lysosomes to cytosol] of the weak organic base acridine orange. We thereafter observed (a) a loss of mitochondrial membrane potential (as from 10 h, based on spectrophotometric and confocal microscopy using JC1 probe) and (b) the release of cytochrome c from granules to cytosol, and the activation of caspase-9 (as from 12 h; evidenced by Western blot analysis). Increase in caspase-3 activity (assayed with Ac-DEVD-AFC in the presence of z-VAD-fmk]) and appearance of fragmented nuclei (DAPI staining) was then detected as from 16 to 24 h together with nuclear fragmentation. Gentamicin produces a fast (within 4 h) release of calcein from negatively-charged liposomes at pH 5.4, which was slowed down by raising the pH to 7.4, or when phosphatidylinositol was replaced by cardiolipin (to mimic the inner mitochondrial membrane). The present data provide temporal evidence that gentamicin causes apoptosis in LLC-PK1 with successive alteration of the permeability of lysosomes, triggering of the mitochondrial pathway, and activation of caspase-3. PMID- 16039944 TI - Inorganic mercury dissociates preassembled Fas/CD95 receptor oligomers in T lymphocytes. AB - Genetically susceptible rodents exposed to low burdens of inorganic mercury (Hg2+) develop autoimmune disease. Previous studies have shown that low, noncytotoxic levels of Hg2+ inhibit Fas-mediated apoptosis in T cells. These results suggest that inhibition of the Fas death receptor pathway potentially contributes to autoimmune disease after Hg2+ exposure, as a consequence of disruption of peripheral tolerance. The formation of active death inducing signaling complexes (DISC) following CD95/Fas receptor oligomerization is a primary step in the Fas-mediated apoptotic pathway. Other recent studies have shown that Hg2+ at concentrations that inhibit apoptosis also inhibit formation of active DISC, suggesting that inhibition of DISC is the mechanism responsible for Hg2+-mediated inhibition of apotosis. Preassociated Fas receptors have been implicated as key elements necessary for the production of functional DISC. We present evidence in this study showing that low and nontoxic concentrations of Hg2+ induce the dissociation of preassembled Fas receptor complexes in Jurkat T cells. Thus, this Hg2+-induced event should subsequently decrease the amount of preassembled Fas available for DISC formation, potentially resulting in the attenuation of Fas-mediated apoptosis in T lymphocytes. PMID- 16039945 TI - Mast cell mediators in citric acid-induced airway constriction of guinea pigs. AB - We demonstrated previously that mast cells play an important role in citric acid (CA)-induced airway constriction. In this study, we further investigated the underlying mediator(s) for this type of airway constriction. At first, to examine effects caused by blocking agents, 67 young Hartley guinea pigs were divided into 7 groups: saline + CA; methysergide (serotonin receptor antagonist) + CA; MK-886 (leukotriene synthesis inhibitor) + CA; mepyramine (histamine H1 receptor antagonist) + CA; indomethacin (cyclooxygenase inhibitor) + CA; cromolyn sodium (mast cell stabilizer) + CA; and compound 48/80 (mast cell degranulating agent) + CA. Then, we tested whether leukotriene C4 (LTC4) or histamine enhances CA induced airway constriction in compound 48/80-pretreated guinea pigs. We measured dynamic respiratory compliance (Crs) and forced expiratory volume in 0.1 s (FEV0.1) during either baseline or recovery period. In addition, we detected histamine level, an index of pulmonary mast cell degranulation, in bronchoalveolar lavage (BAL) samples. Citric acid aerosol inhalation caused decreases in Crs and FEV0.1, indicating airway constriction in the control group. This airway constriction was significantly attenuated by MK-886, mepyramine, cromolyn sodium, and compound 48/80, but not by either methysergide or indomethacin. Both LTC4 and histamine infusion significantly increased the magnitude of CA-induced airway constriction in compound 48/80-pretreated guinea pigs. Citric acid inhalation caused significant increase in histamine level in the BAL sample, which was significantly suppressed by compound 48/80. These results suggest that leukotrienes and histamine originating from mast cells play an important role in CA inhalation-induced noncholinergic airway constriction. PMID- 16039946 TI - Repeated in utero and lactational 2,3,7,8-tetrachlorodibenzo-p-dioxin exposure affects male gonads in offspring, leading to sex ratio changes in F2 progeny. AB - The effects of in utero and lactational 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the reproductive system of male rat offspring (F1) and the sex ratio of the subsequent generation (F2) were examined. Female Holtzman rats were gavaged with an initial loading dose of 400 ng/kg TCDD prior to mating, followed by weekly maintenance doses of 80 ng/kg during mating, pregnancy, and the lactation period. Maternal exposure to TCDD had no significant effects on fetus/pup (F1) mortality, litter size, or sex ratio on gestation day (GD) 20 or postnatal day (PND) 2. The TCDD concentration in maternal livers and adipose tissue on GD20 was 1.21 and 1.81 ng/kg, respectively, and decreased at weaning to 0.72 in the liver and 0.84 in the adipose tissue. In contrast, the TCDD concentration in pup livers was 1.32 ng/kg on PND2 and increased to 1.80 ng/kg at weaning. Ventral prostate weight of male offspring was significantly decreased by TCDD exposure on PND28 and 120 compared with that of controls. Weight of the testes, cauda epididymides, and seminal vesicle, and sperm number in the cauda epididymis were not changed by TCDD exposure at PND120. TCDD- or vehicle-exposed male offspring were mated with unexposed females. The sex ratio (percentage of male pups) of F2 offspring was significantly reduced in the TCDD-exposed group compared with controls. These results suggest that in utero and lactational TCDD exposures affect the development of male gonads in offspring (F1), leading to changes in the sex ratio of the subsequent generation (F2). PMID- 16039947 TI - Different cellular responses evoked by natural and stoichiometric synthetic chrysotile asbestos. AB - The carcinogenic potency of asbestos, including chrysotile, is well established. Several physico-chemical features of the fibers appear implied, such as fibrous habit, size, crystallinity, morphology, and surface active metal ions, where free radical generation may take place. In contrast to other asbestos forms, iron is not a stoichiometric component of chrysotile, but is only present together with other extraneous ions as a magnesium- and silicon-replacing contaminant. To determine the role played by contaminating ions and morphological features of the fibers, a stoichiometric chrysotile with constant structure and morphology was synthesized in hydrothermal conditions. Free radical generation and the effects of these fibers on human lung epithelial A549 cells have been compared to that elicited by a well known toxic natural chrysotile (UICC A, from Rhodesia). After a 24-h incubation, the natural, but not the synthetic, form exerted a cytotoxic effect, detected as leakage of lactate dehydrogenase. Homolytic rupture of a C-H bond and lipoperoxidation in A549 cells took place in the presence of the natural, but not of the synthetic, chrysotile. Antioxidant systems were also affected differently. The pentose phosphate pathway and its regulatory enzyme glucose 6-phosphate dehydrogenase were markedly inhibited only by the natural specimen, which also caused a depletion of intracellular reduced glutathione in A549 cells. These results suggest that metal ions, fiber size and state of the surface play a crucial role in the oxidative stress caused by chrysotile asbestos. Stoichiometric synthetic fibers may thus be proposed as a reference standard (negative control) for toxicological studies. PMID- 16039948 TI - Conditionally MYC: insights from novel transgenic models. AB - MYC was one of the first oncogenes identified to be associated with chromosomal aberrations and one of the most common oncogenes involved in the pathogenesis of cancer. However, until recently it was not clear if MYC would be a good target for the treatment of cancer. New conditional transgenic models have been used to demonstrate that even the brief inactivation of MYC can reverse tumorigenesis. Here we review results from recent experimental model systems, which demonstrate that the inactivation of MYC may be a specific and effective treatment for many types of cancer. PMID- 16039949 TI - Altered protein expression in endometrial carcinogenesis. AB - We have discovered several protein biomarkers that are altered during carcinogenesis of the human uterine endometrium. Proteins prepared from 19 endometrial carcinomas (Group A), and 20 normal endometria obtained from benign gynecological conditions (Group B), were investigated by Surface Enhanced Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (SELDI-TOF-MS). Two proteins, EC1 and EC2, were consistently expressed differentially. EC1 had an increased level of expression in carcinomas (P<0.001), while EC2 was expressed at a lower level (P=0.004). The isoelectric points of EC1 and EC2 were approximately pH 5.0 and 7.0, respectively. These proteins are thus potential biomarkers for detection of endometrial carcinoma. PMID- 16039951 TI - The heterocyclic ruthenium(III) complex KP1019 (FFC14A) causes DNA damage and oxidative stress in colorectal tumor cells. AB - The Ru(III) complex salt KP1019 induced formation of H2O2 in colorectal tumor cells in a dose-dependent way. It also caused DNA-strand breaks if only weakly doubling tail length to 55.87+/-3.97 microm. Both effects were prevented by 5mM N acetylcysteine (NAC) which also reduced cytotoxicity (IC(50) 55 vs 30 microM without NAC). Induction of apoptosis was shown by loss of mitochondrial membrane potential in 63.4+/-2.1% of the population and by caspase-dependent cleavage of poly-(ADP-ribose)-polymerase (PARP). Both effects were inhibited by NAC which reduced the population with depolarized mitochondrial membranes to 24.1+/-1.2% and prevented PARP-cleavage indicating a central role oxidative stress in KP1019 induced apoptosis. PMID- 16039950 TI - Inhibitory effects of Pfaffia paniculata (Brazilian ginseng) on preneoplastic and neoplastic lesions in a mouse hepatocarcinogenesis model. AB - Studies have been demonstrating Pfaffia paniculata root (Brazilian ginseng) anticarcinogenic activities. We evaluated its chemopreventive effects on preneoplastic hepatic lesions. BALB/c aged-15 days received 10mug/g of diethylnitrosamine carcinogen, i.p. They were fed with the powdered root added to the diet: 0.5, 2 or 10% during 27 weeks. After being sacrificed, the macroscopic lesions in the livers were examined. Preneoplastic or neoplastic lesions were measured, quantified and classified morphologically. The treatment reduced the incidence, mean area and number of lesions, indicating an inhibitory effect of these roots on hepatocarcinogenesis promotion or progression steps. PMID- 16039952 TI - Antagonists of growth hormone releasing hormone and bombesin inhibit the expression of EGF/HER receptor family in H-69 small cell lung carcinoma. AB - Effects of in vivo treatment with antagonists of growth hormone-releasing hormone (GHRH), JV-1-65 and MZ-J-7-110, and bombesin/gastrin-releasing peptide antagonist RC-3940-II, on the EGF receptor (EGFR) family, were investigated in H-69 SCLC. Tumors were analyzed by RT-PCR, immunoblotting and binding assays. Treatment with these analogs reduced the binding capacity of EGFR by 18-64%, and inhibited the mRNA expression for EGFR, HER-2 and -3 by 27-75.4, 17-26.3, and 13.8-46.6%, respectively. The antagonists also decreased the protein levels for EGFR by 21 34%, HER-2 by 36-68% and HER-3 by 43-49%. This is the first demonstration that antiproliferative effects of GHRH antagonists are associated with a downregulation of EGF/HER receptors. PMID- 16039953 TI - Phenotypic characterization of chondrosarcoma-derived cell lines. AB - Gene expression profiling of three chondrosarcoma derived cell lines (AD, SM, 105KC) showed an increased proliferative activity and a reduced expression of chondrocytic-typical matrix products compared to primary chondrocytes. The incapability to maintain an adequate matrix synthesis as well as a notable proliferative activity at the same time is comparable to neoplastic chondrosarcoma cells in vivo which cease largely cartilage matrix formation as soon as their proliferative activity increases. Thus, the investigated cell lines are of limited value as substitute of primary chondrocytes but might have a much higher potential to investigate the behavior of neoplastic chondrocytes, i.e. chondrosarcoma biology. PMID- 16039954 TI - pp32/ I-1(PP2A) negatively regulates the Raf-1/MEK/ERK pathway. AB - In this study, we focus on the potential interaction of pp32/I-1(PP2A) (pp32) with the Raf-MEK-ERK signaling pathway. We show that overexpressed pp32 suppresses Raf-1 activation, thereby downregulating the level of ERK activation. This suppression of Raf-1 requires the C-terminal half of pp32. Conversely, knock down of PP32 by siRNA enhances ERK and MEK activations. Taken together, we propose that tumor-suppression by pp32 is, at least in part, mediated by downregulation of the Raf-MEK-ERK signaling pathway. PMID- 16039955 TI - Mechanism of action of potato carboxypeptidase inhibitor (PCI) as an EGF blocker. AB - The epidermal growth factor receptor (EGFR) signal transduction pathway plays a prominent role in the development of carcinomas, and is an interesting target for antitumoral therapy. We have previously described how potato carboxypeptidase inhibitor (PCI), a 39-amino acid protease inhibitor with a T-Knot motif, binds to EGFR receptor and inhibits the activation of receptor protein tyrosine kinase. In this paper it is shown that PCI interferes with EGFR activation through inhibition of receptor dimerization and receptor transphosphorylation induced by epidermal growth factor (EGF) and by transforming growth factor alpha (TGF alpha). Moreover, PCI blocks the formation and activation of ErbB1/ErbB-2 heterodimers that have a prominent role in carcinoma development. As a result of these effects, PCI interferes in the EGFR signal transduction pathway by reversing the effects of EGF on the growth of two tumoral cell lines, A431 and MDA-MB-453, and promotes EGFR down-regulation. These results show that PCI acts as an EGF/TGF-alpha antagonist, which suggests its therapeutic potential in the treatment of carcinomas. PMID- 16039956 TI - Approaches to managing bone metastases from breast cancer: the role of bisphosphonates. AB - Conventional management of metastatic bone disease involves local and systemic therapies in various combinations, along with symptomatic management to provide optimal care. In recent years, it has become clear that adding bisphosphonates to these treatments reduces the incidence and severity of skeletal complications. Bisphosphonates can also relieve metastatic bone pain and improve quality of life, although the extent to which they have demonstrated these effects may differ between agents. While bisphosphonates are the standard of care for the treatment of bone metastases, clinical trials are investigating additional indications for these agents, including the use of intensive dosing regimens for the relief of severe or opioid-resistant metastatic bone pain and adjuvant treatment for the prevention of bone metastases and cancer treatment-induced bone loss. Current and future indications demand effective, well-tolerated and convenient bisphosphonates, and the benefits of different drugs must be balanced against their limitations. The cost-effectiveness of bisphosphonate treatment is also a consideration, given the high economic burden of metastatic bone disease from breast cancer. PMID- 16039957 TI - What knowledge do ICU nurses have with regard to the effects of noise exposure in the Intensive Care Unit? AB - This small-scale study was undertaken to assess what knowledge nursing staff from a General Intensive Care Unit held with regard to noise exposure. To assess knowledge a self-administered multiple-choice questionnaire was used. Rigorous peer-review insured content validity. This study produced poor results in terms of the knowledge nurses held with regard to noise related issues in particular the psychophysiological effects and current legislation concerning its safe exposure. Non-parametric testing, using Kruskal-Wallis found no significant difference between nursing grades, however, descriptive analysis demonstrated that the staff nurse grade (D and E) performed better overall. Whilst the results of this study may seem self-evident in some respects, it is the problems of exposure to excessive noise levels for both patients and hospital personnel, which are clearly not understood. The effects noise exposure has on individuals for example decreased wound healing; sleep deprivation and cardiovascular stimulation must be of concern especially in terms of patient care but more so for nursing staff especially the effects noise levels can have on cognitive task performance. PMID- 16039958 TI - Noise and sleep disturbance factors before and after implementation of a behavioural modification programme. AB - The purpose of this study was to examine sleep disturbance factors documented by the personnel and recorded noise level during two weeks before (measurement one M1) and after (measurement two M2) the implementation of a behavioural modification programme. The behavioural modification programme included non disturbance periods on a neurointensive care unit (NICU) in a University hospital. Sleep disturbance factors were identified as general and specific nursing care, and medical treatment documented over fourteen 24-h periods. Minimum, maximum and peak mean noise levels in dBA were recorded continuously during fourteen 24-h periods by a decibel meter. The behavioural modified programme included changing nursing and medical routines and the introduction of afternoon and night non-disturbance periods. The most disturbing documented factors at both M1 and M2 were general nursing care activities. Noise levels showed great variation at both M1 and M2. At M2 minimum and maximum noise levels varied when compared to M1. Implementation of a behavioural modification programme and non-disturbance periods co-ordinated routines resulted in reduced sleep disturbance factors and partly reduced noise levels on the NICU. Changes of the physical care/working environment, preparations before non-disturbance periods, regular evaluations of routines and education are needed to improve sleep on NICUs. PMID- 16039959 TI - Designed sound and music environment in postanaesthesia care units--a multicentre study of patients and staff. AB - A multicentre study in five postanaesthesia care units (PACUs) was performed to investigate patient and staff opinion of a specially designed music environment (DME), related to geographical location. Patients (325) and staff (91) described their opinion by means of a questionnaire-anonymously in the case of staff. Patients were not asked beforehand for permission to play music. Amongst patients 267 (83%) found the sound environment with DME pleasant or very pleasant, 26 (6%) found it unpleasant, whereas 32 (11%) answered "no opinion". The opinion of the patients did not differ significantly with geographical location. A strong correlation (P<0.05) between a positive attitude towards DME and degree of relaxation and satisfaction with stay was found. The staff had an equally positive attitude towards the DME; but theirs varied significantly with location. The opinion of the staff was more similar concerning the beneficial effect on working conditions and distress, but varied still significantly. The opinion of the staff had no demonstrable impact on that of the patients. PMID- 16039960 TI - Organ and tissue donation: a survey of nurse's knowledge and educational needs in an adult ITU. AB - The aim of this paper is to present the results of a survey that was undertaken to assess nurses' knowledge and educational needs towards organ donation within one adult general intensive care unit. The survey consists of 31 registered nurses who completed a confidential questionnaire that aimed to assess their existing knowledge and deficits in organ and tissue donation. The survey highlights the sample lacked confidence in approaching relatives for donation consent, deficits in brain stem death testing and donor criteria. It was also apparent that a significant number of nurses could not identify which tissues can be donated and the contraindications for tissue donation. A majority of the sample stated their knowledge of donation issues would improve if an educational programme were developed on organ donation. This is further supported by previous work by [Bidigare S, Oermann M, 1991. Attitudes and knowledge of nurses regarding organ procurement. Heart & lung 1:20-3; Smith-Brew S, Yanai L, 1996. The organ donation process through a review of the literature. Part 1. Accident & emergency nursing 4:5-11; Roark D, 2000. Overhauling the organ donation system. Am J Nurs 6:44-9] who suggest that educational programmes covering donation issues should enhance nurses' knowledge and confidence in the organ donation process and ultimately increase the number of potential donors. PMID- 16039962 TI - Non-invasive ventilation (NIV). An audit across the North Central London Critical Care Network (NCLCCN). AB - This NIV audit was conducted across the North Central London network area, for a 3-month period in 2003. Five trusts participated (on 6 hospital sites) and 141 audit forms were returned, allowing data analysis for individual hospitals and for comparisons between hospitals. The results are discussed in line with the British Thoracic Society guidelines on NIV [British Thoracic Society Guideline. Non-invasive ventilation in acute respiratory failure. Thorax 2002;57:192-211] and the Modernisation Agency document [Critical Care Programme. Weaning and Long Term Ventilation. NHS Modernisation Agency; 2002]. Key areas for future work have been identified to facilitate service improvement. PMID- 16039961 TI - Closed versus partially ventilated endotracheal suction in extremely preterm neonates: physiologic consequences. AB - This randomized cross over study aimed to compare the severity and incidences of desaturation and bradycardia between the partially ventilated endotracheal suction method (PVETS) and closed tracheal suction system (CTSS) in extremely preterm neonates. Fifteen intubated and ventilated extremely low birth weight preterm infants (mean birth weight 689g) randomly underwent both suction techniques within a 12-h period to obtain a paired reading group. The process was repeated 24-48h apart until three pairs of reading groups were collected. Changes in oxygen saturation measured with pulse oximetry and heart rate changes measured with electrocardiogram were recorded using Hewlett-Packard m240A monitor trending software. The mean of each parameter's variation from baseline was obtained using SPSS descriptive statistics and analyzed using SPSS repeated measures ANOVA. Fisher Exact Test was used to analyze the incidence of desaturation and bradycardia. The closed tracheal suction system reported a significantly smaller degree of oxygen saturation fall (P<0.005) and significantly fewer incidences of desaturation. There was also a significantly smaller degree of heart rate reduction although episodes of bradycardia were not significantly different between the two methods. Oxygen saturation and heart rate were significantly more stable during the use of CTSS compared to PVETS in the extremely low birth weight preterm population. PMID- 16039963 TI - Up to 16 years follow-up of aortic valve reconstruction with pericardium: a stentless readily available cheap valve? AB - OBJECTIVE: There is lack of information regarding the long-term behavior of aortic valve reconstruction with pericardium (AoR). A 16-year follow-up is reported here. METHODS: Between 1988 and 1995, 92 consecutive patients had AoR with bovine (Group I, n=27) or glutaraldehyde-treated autologous pericardium (Group II, n=65). The mean age was 30 years (range 12-68). There were 65% males, 92% in sinus rhythm, 84% had rheumatic etiology and 36% had 'other valve' surgery. Mitral valve replacement with a mechanical prosthesis is a contraindication to the operation. RESULTS: Hospital mortality was 2%. The reconstructed aortic valve performed well with excellent hemodynamics. The mean follow-up interval was 10.5+/-4 years, range 9-16 years (longer for group I,12 versus 10 years) with 4% late deaths and seven patients lost to follow-up. Survival rate was 85+/-4%. There were no episodes of thromboembolism. Freedom from reoperation for the whole group was 68+/-5% at 10 years and 47+/-6% at 16 years. For group I, it was 68+/-9% at 10 years and 48+/-10% at 16 years, while for group II it was 72+/-6 and 45+/-8% at 10 and 15 years, respectively. Excluding endocarditis (one in group I and seven in group II) and 'other' reasons for reoperation (two in group I and three in group II), the freedom from structural valve degeneration (SVD) at 10 and 16 years was 78+/-1 and 55+/-10% for group I. For group II, it was 80+/-5% at 10 years and 58+/-9% at 15 years. The mean interval at which the valve degenerated was 8.8 years+/-3.6 and did not differ between the two groups. CONCLUSIONS: AoR is feasible with good hemodynamics, low mortality and thromboembolic rate. Its behavior at 10 years is comparable to that of stentless aortic valve bioprosthesis. It can be performed with either xenopericardium or glutaraldehyde-treated autologous pericardium, but the latter has the advantage of being inexpensive and readily available. PMID- 16039964 TI - Brachiocephalic artery to left innominate vein fistula following median sternotomy. PMID- 16039965 TI - Secondary elephant trunk fixation with endovascular stent grafting for extensive/multiple thoracic aortic aneurysm. AB - Four patients who underwent secondary elephant trunk fixation by endovascular stent grafting are presented and the advantage of this method to treat multiple/extensive thoracic aortic aneurysm is discussed. In two of them, the elephant trunk installation has been performed at another hospital for extensive aortic aneurysm. In two other patients, the aortic arch replacement and the elephant trunk installation were performed through median sternotomy, initially for multiple aortic lesions, including both arch and descending aorta. No neurological deficit, stroke nor spinal cord injury was encountered during the follow-up period (24-40 months). The diameter of the aneurysms decreased markedly in three patients. In one patient, the aneurysm expanded gradually and type II endoleak was treated by coil embolization. In one patient, who showed marked shrinkage of the aneurysm, the stent graft kinked mildly. Based on the low mortality rate of well-established aortic arch surgery, concomitant elephant trunk installation which was followed by the secondary fixation with endovascular stent grafting might be useful to treat multiple/extensive thoracic aneurysm from distal arch to descending aorta. PMID- 16039966 TI - PPARgamma: a critical determinant of body fat distribution in humans and mice. AB - Evidence emerging from studies of humans and mice has indicated peroxisome proliferator-activated receptor gamma (PPARgamma) to be not only a key factor for adipogenesis but also a critical determinant of body fat distribution. Whereas genetically reduced PPARgamma activity in adipose tissue leads to reduction of total fat mass in humans and in mice, mutations in the ligand-binding domain of PPARgamma cause abnormal body fat distributions. It is less clear from mutation analysis how PPARgamma is involved in metabolic disturbances such as insulin resistance and its cardiovascular complications. Nevertheless, similarities and differences in the phenotypes associated with PPARgamma mutations in humans and in mouse models provide opportunities to dissect relationships between body fat distribution and its metabolic complications. PMID- 16039967 TI - Platelet collagen receptors and coagulation. A characteristic platelet response as possible target for antithrombotic treatment. AB - Collagen is a unique agonist of platelets, because it acts as an immobilized ligand that only causes platelet activation after stable adhesion. This review addresses the present understanding of how platelet interaction with collagen supports the process of thrombin generation and coagulation. Only some of the collagen-adhered platelets, that is, those showing profound changes in shape and shedding microparticles (resembling apoptotic cells), appear to contribute to the procoagulant activity of platelets. The main signaling receptor for collagen, glycoprotein VI, plays a key role in the platelet procoagulant response during thrombus formation; this is a reason why new anti-glycoprotein-VI antibodies are promising antithrombotic tools. PMID- 16039968 TI - Caveolae, lipid rafts, and vascular disease. AB - Caveolae and lipid rafts are discrete regions within the plasma membrane that coordinate and regulate a variety of signaling processes. The exact relationship between caveolae and lipid rafts is unclear. However, caveolae contain a protein called caveolin that serves as a biochemical marker for caveolae. In addition, caveolin plays a role in maintaining the lipid composition of caveolae, the morphology of caveolae, and the signals that emanate from caveolae. The physiologic importance of caveolae is evidenced by recent studies using caveolin knockout mice that show dramatic abnormalities in the cardiovascular system, such as pulmonary hypertension and cardiac hypertrophy. In this review, we will focus on the role of caveolae in the cardiovascular system. PMID- 16039969 TI - A role for MSX2 and necdin in smooth muscle differentiation of mesoangioblasts and other mesoderm progenitor cells. AB - The molecular regulation of smooth muscle differentiation is currently far less well understood than that of striated muscle, in part because in this cell type, the differentiated state is plastic and reversible. In recent years, however, several molecules, the best characterized of which is myocardin, have been shown to be necessary and sufficient to promote at least partial smooth muscle differentiation. Indeed, mice deficient in myocardin have a severe reduction of smooth muscle tissue. However, possibly because of multiple embryological origins, which include mesenchyme, neural crest, and even endothelium, different types of smooth muscle cells differ in their expression of myocardin and of other potential regulatory molecules. Here, we will review recent work on the topic, focusing on the mesoangioblast, a recently described vessel-associated stem cell, whose differentiation into smooth muscle is dependent upon expression of msx2 and necdin, but not of myocardin. PMID- 16039970 TI - Mechanisms of cell survival in myocardial hibernation. AB - Myocardial hibernation represents a condition of regional ventricular dysfunction in patients with chronic coronary artery disease, which reverses gradually after revascularization. The precise mechanism mediating the regional dysfunction is still debated. One hypothesis suggests that chronic hypoperfusion results in a self-protecting downregulation in myocardial function and metabolism to match the decreased oxygen supply. An alternative hypothesis suggests that the myocardium is subject to repetitive episodes of ischemic dysfunction resulting from an imbalance between myocardial metabolic demand and supply that eventually creates a sustained depression of contractility. It is generally agreed that hibernating myocardium is submitted repeatedly to ischemic stress, and therefore one question persists: how do myocytes survive in the setting of chronic ischemia? The hallmark of hibernating myocardium is a maintained viability of the dysfunctional myocardium which relies on an increased uptake of glucose. We propose that, in addition to this metabolic adjustment, there must be molecular switches that confer resistance to ischemia in hibernating myocardium. Such mechanisms include the activation of a genomic program of cell survival as well as autophagy. These protective mechanisms are induced by ischemia and remain activated chronically as long as either sustained or intermittent ischemia persists. PMID- 16039971 TI - AMP-activated protein kinase: a key stress signaling pathway in the heart. AB - AMP-activated protein kinase (AMPK) is activated during exercise and ischemia and is emerging as an important regulatory mechanism in the heart. AMPK promotes adenosine triphosphate-generating pathways, including glucose transport, glycolysis, and fatty acid oxidation, while inhibiting energy-consuming anabolic pathways. After ischemia-reperfusion, AMPK-deficient hearts from transgenic mice have severe left ventricular contractile dysfunction with increased apoptosis and necrosis. Mutations in the AMPKgamma(2) subunit lead to cardiac glycogen overload, Wolff-Parkinson-White syndrome, arrhythmias, and heart failure. This review focuses on the molecular mechanisms of activation and cardiovascular actions of AMPK in the heart. PMID- 16039972 TI - The mitochondrial biogenesis regulatory program in cardiac adaptation to ischemia -a putative target for therapeutic intervention. AB - The resurgence of mitochondrial biology research stems from the realization that the distinct regulation of mitochondria to meet diverse homeostatic demands is driven by exquisite biochemical and molecular control mechanisms. This program termed mitochondrial biogenesis is integral to orchestrating mitochondrial function and appears to exhibit adaptive remodeling following biomechanical and oxidative stress. The major bioenergetic function of mitochondria partitions the final utilization of oxygen between oxidative phosphorylation and reactive oxygen species. As disruption in oxidative phosphorylation and excessive reactive oxygen species contribute to cardiac ischemia-reperfusion injury, we hypothesize that the mitochondrial biogenesis regulatory program is an explicit target for cardiac therapeutic interventions. The objectives of this review are to (a) define the advances in understanding the mitochondrial biogenesis regulatory program integrated to its control of mitochondrial bioenergetics and oxygen utilization, (b) reveal how this program is modulated by chronic hypoxia and ischemic preconditioning, and (c) examine the therapeutic potential of modulating the regulation of mitochondrial biogenesis as a strategy to attenuate ischemia reperfusion injury. PMID- 16039973 TI - Imaging exams of bone lesions in patients with diffuse cutaneous leishmaniasis (DCL). AB - We studied bone lesion alterations in three patients with diffuse cutaneous leishmaniasis (DCL) by imaging exams (radiography and scintigraphy) and histopathology. Two patients had bone lesions of distal extremities of hands and feet, and one infiltrating plaques in the skin. The study was conducted at three specialized centers (Presidente Dutra Hospital/Nucleus of Tropical Pathology, UFMA-MA; Goncalo Moniz Research Center-FIOCRUZ-BA; Laboratory of Pathology of Infectious Diseases (LIM-50), University of Sao Paulo, SP). Three-phase bone scintigraphy demonstrated high sensitivity and specificity for bone lesions, showing increased uptake of the radiopharmaceutical at sites of active lesions. In contrast, radiography demonstrated lytic lesions, cortical destruction and local osteopenia of the bone extremeties in two patients. Histopathological analysis showed sequestration with presence of amastigote forms of Leishmania (osteomyelitis), mononuclear cells and macrophages containing amastigote forms of Leishmania in one patient. These preliminary data indicate that imaging exams (radiography and scintigraphy) are important in the evaluation of bone lesions in diffuse cutaneous leishmaniasis and should be included in the routine histopathological diagnosis of the disease and follow-up of bone lesions. PMID- 16039974 TI - The construction of DNA molecules of figure-eight structure. AB - Using DNA molecules to construct a structural scaffold for nanotechnology is largely accepted. In this article, we report on two methods for constructing a figure-eight structure of DNA molecules having a relatively high yield that could be used further as a scaffold for nanotechnology applications. In the first method, two plasmids were constructed that, on digestion with a restriction endonuclease producing nicks in the corresponding sites and after heating, produced complementary single-stranded sequences, enabling the plasmids to hybridize to each other and forming a figure-eight structure. The formation of the figure-eight structure was analyzed by restriction analysis and gel electrophoresis as well as by atomic force microscopy. The second method makes use of the bacteriophage M13 that is obtained as either a single- or double stranded circular DNA molecule. Two M13 molecules harboring complementary sequences were constructed and produced a figure-eight structure on hybridization. The methods described here could be used further for the construction of nanoelectronic devices. PMID- 16039975 TI - Solid-phase extraction as a tool to remove impurities and small fragments from synthetic peptides. PMID- 16039976 TI - Fluorometric assay for detection and quantitation of polyamidoamine dendrimers. AB - Polyamidoamine dendrimers are protein-like nanomaterials that have many potential biological applications. Research studies using dendrimers often require their detection and quantitation. This article describes a simple, inexpensive, and rapid fluorometric assay based on the ability of dendrimers to bind the fluorescent probe, 8-anilino-1-naphthalene-sulfonic acid. This interaction leads to an enhanced fluorescence intensity of the dye that shows a linear relationship with dendrimer concentration. The procedure is applicable to dendrimers that have various surface functional groups and internal cores. Amine- and hydroxyl terminated dendrimers gave the best calibration curves under alkaline conditions (pH 7.4). Carboxyl-terminated dendrimers required acidic conditions for their measurement. The assay for dendrimers with an amine surface and an ethylene diamine core was accurate between 0 and 100 microM and produced a within-run coefficient of less than 5%. Under the assay conditions used, the fluorescence signal strength was dependent on type of surface group and core length of the dendrimer. PMID- 16039977 TI - Direct electrochemistry and electrocatalytic activity of catalase incorporated onto multiwall carbon nanotubes-modified glassy carbon electrode. AB - The direct voltammetry and electrocatalytic properties of catalase, which was adsorbed on the surface of multiwall carbon nanotubes (MWCNTs), was investigated. A pair of well-defined and nearly reversible cyclic voltammetry peaks for Fe(III)/Fe(II) redox couple of catalase adsorbed on the surface of MWCNTs at approximately -0.05 V versus reference electrode in pH 6.5 buffer solution, indicating the direct electron transfer between catalase and electrode. The surface coverage of catalase immobilized on MWCNTs glassy carbon electrode was approximately 2.4x10(-10) molcm-2. The transfer coefficient (alpha) was calculated to be 0.4, and the heterogeneous electron transfer rate constant was 80 s-1 in pH 7, indicating great facilitation of the electron transfer between catalase and MWCNTs adsorbed on the electrode surface. The formal potential of catalase Fe(III)/Fe(II) couple in MWCNTs film had a linear relationship with pH values between 2 and 11 with a slope of 58 mV/pH, showing that the electron transfer is accompanied by single proton transportation. Catalase adsorbed on MWCNTs exhibits a remarkable electrocatalytic activity toward the reduction of oxygen and hydrogen peroxide. The value for calculated Michaelis-Menten constant (1.70 mM) was high, indicating the potential applicability of the films as a new type of reagentless biosensor based on the direct electrochemistry of the catalase enzyme. PMID- 16039978 TI - Archived gels as a tool for identification of protein complexes: Polo kinase cofractionates with Drosophila 205-kDa MAP and ncd in mitotic embryonic extracts. PMID- 16039979 TI - Determination of hydrogen peroxide generated by reduced nicotinamide adenine dinucleotide oxidase. AB - Hydrogen peroxide can be conveniently determined using horseradish peroxidase (HRP) and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid). However, interference occurs among assay components in the presence of reduced nicotinamide adenine dinucleotide (NADH) that is also a substrate of NADH oxidase. So, depletion of NADH is required before using the HRP method. Here, we report simple and rapid procedures to accurately determine hydrogen peroxide generated by NADH oxidase. All procedures developed were based on the extreme acid lability of NADH and the stability of hydrogen peroxide, because NADH was decomposed at pH 2.0 or 3.0 for 10 min, while hydrogen peroxide was stable at pH 2.0 or 3.0 for at least 60 min. Acidification and neutralization were carried out by adjusting sample containing NADH up to 30 microM to pH 2.0 for 10 min before neutralizing it back to pH 7.0. Then, hydrogen peroxide in the sample was measured using the HRP method and its determination limit was found to be about 0.3 microM. Alternatively, hydrogen peroxide in samples containing NADH up to 100 microM could be quantitated using a modified HRP method that required an acidification step only, which was found to have a determination limit of about 3 microM hydrogen peroxide in original samples. PMID- 16039980 TI - Application of Alamar blue/5-carboxyfluorescein diacetate acetoxymethyl ester as a noninvasive cell viability assay in primary hepatocytes from rainbow trout. AB - We have adopted the application of two fluorescent indicator dyes to studying the viability of monolayers of primary rainbow trout hepatocytes. The two fluorescent dyes--Alamar blue, which indicates metabolic activity of a cell, and 5 carboxyfluorescein diacetate acetoxymethyl ester (CFDA-AM), which is an indirect measure of cell membrane integrity-are noninvasive and can be monitored conveniently directly in multiwell plates. According to these dyes, L-15 culture medium supported hepatocyte viability over 96 h more stably than did M199. The two dyes proved to be capable of detecting a concentration-dependent toxic insult to hepatocytes caused by the model compound, pentachlorophenol. In contrast, a lack of impact on cell viability was indicated for up to 10(-5) M 17beta estradiol, and that observation was supported by the induction of vitellogenin (VTG) mRNA/protein as indicator of hepatocyte-differentiated function. Application of the Alamar blue/CFDA-AM for 30 min did not alter gene expression either specifically as reflected by VTG or generally as reflected by a random selection of gene sequences that were amplified by differential display reverse transcription PCR (dd-rt-PCR). Thus, the assay represents a resource-efficient way of integrating measures of cell viability and gene expression that should aid in the interpretation of in vitro results. The assay can be applied repeatedly to the same set of cells and can be performed just prior to analysis of gene expression. PMID- 16039981 TI - Properties of nonfused liposomes immobilized on an L1 Biacore chip and their permeabilization by a eukaryotic pore-forming toxin. AB - The L1 chip is used intensively for protein-membrane interaction studies in Biacore surface plasmon resonance systems. The exact form of captured lipid membranes on the chip is, however, not precisely known. Evidence exists that the vesicles both remain intact after the binding to the chip and fuse to form a large single-bilayer membrane. In this study, we were able to bind up to approximately 11,500 resonance units of zwitterionic liposomes (100 nm in diameter) at a low flow rate. We show by fluorescence microscopy that the entire surface of the flow cell is covered homogeneously by liposomes. Negatively charged vesicles (i.e., those composed of phosphatidylcholine/phosphatidylglycerol [1:1]) always deposited less densely, but we were able to increase the density slightly with the use of calcium chloride that promotes fusion of the vesicles. Finally, we used zwitterionic liposomes loaded with fluorescent probe calcein to show that they remain intact after the capture on the L1 chip. The fluorescence was lost only after we used equinatoxin, a well-studied pore-forming toxin, to perform on-chip permeabilization of vesicles. The characteristics of permeabilization process for chip-immobilized liposomes are similar to those of liposomes free in solution. All results collectively suggest that liposomes do not fuse to form a single bilayer on the surface of the chip. PMID- 16039982 TI - Mechanisms of selenium inhibition of cell apoptosis induced by oxysterols in rat vascular smooth muscle cells. AB - Our previous study reported that oxysterol cholestane-3beta,5 alpha, 6 beta-triol (Triol) induced vascular smooth muscle cells (VSMCs) apoptosis, which was inhibited by selenium pretreatment. To further investigate the mechanisms of the inhibition, the glutathione peroxidase (GPx) activity, the total antioxidant capacity (T-AOC), the total superoxide dismutase (SOD) activity, and the level of lipid peroxidation (the content of malondialdehyde, MDA) of VSMCs were measured, and fluidity of cell membrane, reactive oxygen species (ROS) level, the reduction of mitochondrial membrane potential (Delta psi(m)), and the intracellular Ca(2+) in single cell were detected using several fluorescence indicators. Meanwhile, the mRNA levels of c-myc, bcl-2, GPx, and thioredoxin reductase (TR) were measured by reverse transcriptase polymerase chain reaction (RT-PCR) analysis. The results showed that the decrease of GPx activity, T-AOC, SOD activity, the fluidity of cell membrane, the Delta psi(m), and the mRNA expression of c-myc, bcl-2, GPx, and TR of VSMCs and the increase of MDA, ROS generation, and intracellular Ca(2+), significantly induced by Triol (10 microM, 24h) were inhibited to a different extent, respectively, when cells were pretreated with sodium selenite (50 nM, 12 or 24h) before exposure to Triol. These effects were time dependent and enhanced with prolongation of the time of pretreatment. In conclusion, the results in the present work showed that the mechanism of selenium inhibition of cell apoptosis induced by oxysterols in rat VSMCs was related with the antioxidation of selenoproteins. PMID- 16039983 TI - Evaluation of the relationship between caries indices and salivary secretory IgA, salivary pH, buffering capacity and flow rate in children with Down's syndrome. AB - The aim of this study was to compare the caries prevalence and salivary secretory IgA (sIgA), salivary pH, buffering capacity and flow rate between Down's syndrome (DS) and control subjects. Seventy-three institutionalised children with DS and 70 normal children aged 7-12 years old were included in this study. Tooth brushing habits and daily dietary sugar exposures of the children, family income and education levels of the parents were recorded. DMFS and dfs scores were assessed according to the World Health Organisation's criteria and stimulated whole saliva samples were collected. Salivary sIgA levels were determined by radial immunodiffusion technique, the average salivary flow rate was measured from the total volume, and salivary pH and buffering capacity were determined using a pH micro-electrode. All data were analysed using SPSS version 11.0. The DMFS and dfs scores were significantly lower in the DS group than the control group (P < 0.05). Otherwise, the difference in plaque scores between the DS and control groups was not statistically significant (P > 0.05). There were no significant differences in tooth-brushing habits and daily dietary sugar exposures of the children, family income and education levels of the parents between the two groups (P > 0.05). Salivary sIgA levels were significantly higher in the DS group (P < 0.05). Salivary pH, buffering capacity and flow rate were quite similar in both the DS and control groups (P > 0.05). In conclusion, the patients with DS had a significantly lower prevalence of caries and significantly higher levels of salivary sIgA in this study. This finding tends to support the hypothesis that higher levels of salivary sIgA may protect against dental caries. PMID- 16039984 TI - Mechanism of inhibition of proton: dipeptide co-transport during chronic enteritis in the mammalian small intestine. AB - Amino acids, a critical energy source for the intestinal epithelial cells, are more efficiently assimilated in the normal intestine via peptide co-transporters such as proton:dipeptide co-transport (such as PepT1). Active uptake of a non hydrolyzable dipeptide (glycosarcosine) was used as a substrate and PepT1 was found to be present in normal villus, but not crypt cells. The mRNA for this transporter was also found in villus, but not crypt cells from the normal rabbit intestine. PepT1 was significantly reduced in villus cells also diminished in villus cell brush border membrane vesicles both from the chronically inflamed intestine. Kinetic studies demonstrated that the mechanism of inhibition of PepT1 during chronic enteritis was secondary to a decrease in the affinity of the co transporter for the dipeptide without an alteration in the maximal rate of uptake (Vmax). Northern blot studies also demonstrated unaltered steady state mRNA levels of this transporter in the chronically inflamed intestine. Proton dipeptide transport is found in normal intestinal villus cells and is inhibited during chronic intestinal inflammation. The mechanism of inhibition is secondary to altered affinity of the co-transporter for the dipeptide. PMID- 16039985 TI - TG1019/OXE, a Galpha(i/o)-protein-coupled receptor, mediates 5-oxo eicosatetraenoic acid-induced chemotaxis. AB - We have previously identified a Galpha(i/o)-protein-coupled receptor (TG1019/OXE) using 5-oxo-6E,8Z,11Z,14Z-eicosatetraenoic acid (5-oxo-ETE) as its ligand. We investigated signal transduction from TG1019 following stimulation with 5-oxo-ETE and role of TG1019 in 5-oxo-ETE-induced chemotaxis, using Chinese hamster ovary cells expressing TG1019 (CHO/TG1019 cells). 5-Oxo-ETE induced intracellular calcium mobilization and rapid activation of MEK/ERK and PI3K/Akt pathways in CHO/TG1019 cells. CHO/TG1019 cells stimulated with 5-oxo-ETE and other eicosanoids exhibited chemotaxis with efficacies related to agonistic activity of each eicosanoid for TG1019. Pretreatment of the cells with pertussis toxin, a phospholipase C (PLC) inhibitor (U73122) or a PI3K inhibitor (LY294002), markedly suppressed 5-oxo-ETE-induced chemotaxis, whereas pretreatment with a MEK inhibitor (PD98059) had no significant effect on the chemotaxis. Our results show that TG1019 mediates 5-oxo-ETE-induced chemotaxis and that signals from TG1019 are transduced via Galpha(i/o) protein to PLC/calcium mobilization, MEK/ERK, and PI3K/Akt, among which PLC and PI3K would play important roles in the chemotaxis. PMID- 16039986 TI - Extensive modulation of a set of microRNAs in primary glioblastoma. AB - MicroRNAs (miRNAs) are short non-coding RNA molecules playing regulatory roles in animals and plants by repressing translation or cleaving RNA transcripts. The specific modulation of several microRNAs has been recently associated to some forms of human cancer, suggesting that these short molecules may represent a new class of genes involved in oncogenesis. In our study, we examined by microarray the global expression levels of 245 microRNAs in glioblastoma multiforme, the most frequent and malignant of primary brain tumors. The analysis of both glioblastoma tissues and glioblastoma cell lines allowed us to identify a group of microRNAs whose expression is significantly altered in this tumor. The most interesting results came from miR-221, strongly up-regulated in glioblastoma and from a set of brain-enriched miRNAs, miR-128, miR-181a, miR-181b, and miR-181c, which are down-regulated in glioblastoma. PMID- 16039987 TI - MYND domain specific interaction of the melanin-concentrating hormone receptor 1 interacting zinc-finger protein with alpha- and beta-tubulin. AB - MIZIP was originally identified as a highly conserved zinc-finger protein from human brain interacting with the C-terminus of the melanin-concentrating hormone receptor 1. However, the cellular functions of MIZIP are still not known. Here, we focussed on the identification of associated proteins using affinity purification from human cells. This resulted in the identification of alpha- and beta-tubulin. The interaction was confirmed in vitro and in vivo using GST pull down and immunoprecipitation assays, and was mapped to the MYND zinc-finger of MIZIP and to the N-terminus of tubulin. Immunoprecipitation and immunocytochemistry analyses demonstrate that MIZIP binds to tubulin but not to cellular microtubules in vivo and that ectopic expression of MIZIP does not interfere with the overall structure of the microtubular cytoskeleton. Our results suggest that MIZIP might play an important role in mammalian cells by associating with tubulin and thus might provide a link between MCHR1 and tubulin functions. PMID- 16039988 TI - Decreased expression of Hsp27 and Hsp70 in transformed lymphoblastoid cells from patients with spinocerebellar ataxia type 7. AB - Spinocerebellar ataxia type 7 (SCA7) is caused by an expansion of unstable CAG repeats within the coding region of the novel gene, ataxin-7, on chromosome 3. This disease is also associated with an accumulation of abnormal proteins, including expanded polyglutamine-containing proteins, molecular chaperones, and the ubiquitin-proteasome system. In this study, two SCA7 lymphoblastoid cell lines (LCLs) with 100 and 41 polyglutamine repeats were utilized to examine the effects of polyglutamine expansion on heat shock proteins. Interestingly, under basal conditions, Western blot and immunocytochemical analysis showed a significant decrease of Hsp27 and Hsp70 protein expression in cells containing expanded ataxin-7, as compared with that of the normal LCL. On the other hand, the protein levels of Hsp60 and Hsp90 were not significantly altered in the mutant LCLs. Results from semi-quantitative RT-PCR indicated that the differences in Hsp70 protein levels were due to transcriptional defects while the reduction of Hsp27 in the mutant cells was not caused by transcriptional defects. Our results further demonstrated that despite of defective protein expression of Hsp27 and Hsp70, a normal heat shock response was observed in lymphoblastoid cells expressing mutant ataxin-7. Taken together, our results indicated that expanded ataxin-7 that leads to neurodegeneration significantly impaired the expression of Hsp27 and Hsp70 protein, which may be, at least in part, responsible for the toxicity of mutant ataxin-7 proteins and ultimately resulted in an increase of stress-induced cell death. PMID- 16039989 TI - Regulation of volume-sensitive Cl- channels in multi-drug resistant MCF7 cells. AB - The P-glycoprotein (P-gp) is thought to be involved in the regulation of volume sensitive chloride channels. In this study, the possible coupling between P-gp and swelling-activated chloride channels has been examined in MCF7 cells with sensitive (MDR-), resistant (MDR+), and reversed resistant (MDR(REV)) phenotypes. Western blot analysis showed that incubation of cells with doxorubicin induced P gp expression in a reversible manner. Exposure of MDR+ cells to hypotonicity resulted in an inhibition of P-gp activity while hypotonic challenges induced swelling-activated chloride currents (I(Cl-swell)) in MDR-, MDR+, and MDR(REV) MCF7 cells. While verapamil inhibited I(Cl-swell) in all cell types, doxorubicin and vincristine rapidly and reversibly inhibited I(Cl-swell) uniquely in MDR+. Intracellular dialysis of MDR+ cells with C219 anti-P-gp antibody abolished the sensitivity of I(Cl-swell) to doxorubicin and led to a response pattern very close to that of MDR- cells. Taken together, these results strongly suggest that the P-glycoprotein regulates I(Cl-swell) in resistant MCF7. PMID- 16039990 TI - hKSR-2 inhibits MEKK3-activated MAP kinase and NF-kappaB pathways in inflammation. AB - Kinase suppressor of ras (KSR) and MEKK3 (MAP kinase kinase kinase) are integral members of the MAP kinase pathway. We have recently identified a new isoform of the KSR family named human kinase suppressor of ras-2 (hKSR-2), and demonstrated that hKSR-2 negatively regulates Cot, a MAP3K family member which is important in inflammation and oncogenesis [P.L. Channavajhala, L. Wu, J.W. Cuozzo, J.P. Hall, W. Liu, L.L. Lin, Y. Zhang, J. Biol. Chem. 278 (2003) 47089-47097]. In this report, we provide evidence that hKSR-2 also regulates the activity of MEKK3 (another MAP3K family member) in HEK-293T cells. We demonstrate that hKSR-2 is a negative regulator of MEKK3-mediated activation of MAP kinase (specifically ERK and JNK) and NF-kappaB pathways, and concurrently inhibits MEKK3-mediated interleukin-8 production. We find that while hKSR-2 blocks MEKK3 activation, it has little to no effect on other members of the MAP3K family, including MEKK4, TAK1, and Ras-Raf, suggesting that its effects are selective. PMID- 16039991 TI - Kinetics of pyrophosphate-driven proton uptake by acidocalcisomes of Leptomonas wallacei. AB - In this work, we show the kinetics of pyrophosphate-driven H+ uptake by acidocalcisomes in digitonin-permeabilized promastigotes of Leptomonas wallacei. The vacuolar proton pyrophosphatase activity was optimal in the pH range of 7.5 8.0, was inhibited by imidiodiphosphate, and was completely dependent on K+ and PPi. H+ was released with the addition of Ca2+, suggesting the presence of a Ca2+/H+ antiport. In addition, X-ray elemental mapping associated with energy filtering transmission electron microscopy showed that most of the Ca, Na, Mg, P, K, Fe, and Zn were located in acidocalcisomes. L. wallacei immunolabeled with antibodies against Trypanosoma cruzi pyrophosphatase show intense fluorescence in cytoplasmatic organelles of size and distribution similar to the acidocalcisomes. Altogether, the results show that L. wallacei acidocalcisomes possess a H+ pyrophosphatase with characteristics of type I V-H+-PPase. However, we did not find any evidence, either for the presence of H+-ATPases or for Na+/H+ exchangers in these acidocalcisomes. PMID- 16039992 TI - The beta-subunit of G proteins is a substrate of protein histidine phosphatase. AB - Increasing evidence suggests that reversible phosphorylation of histidine residues in proteins is important for signaling cascades in eukaryotic cells. Recently, the first eukaryotic protein histidine phosphatase (PHP) was identified. The beta1-subunit of heterotrimeric G proteins (Gbeta) undergoes phosphorylation on His266 which is apparently involved in receptor-independent G protein activation. We studied whether phosphorylated Gbeta-subunits are substrates of PHP. Phosphorylated Gbetagamma dimers of the retinal G protein transducin and Gbeta in membrane preparations of H10 cells (neonatal rat cardiomyocytes) were dephosphorylated by PHP. Overexpression of PHP in H10 cells showed that PHP and Gbeta also interfere within cells. In membranes of cells overexpressing PHP, the amount of phosphorylated Gbeta was largely reduced. Both our in vitro and cell studies indicate that phosphorylated Gbeta-subunits of heterotrimeric G proteins are substrates of PHP. Therefore, PHP might play a role in the regulation of signal transduction via heterotrimeric G proteins. PMID- 16039993 TI - Role of PYK2 in the development of obesity and insulin resistance. AB - Non-receptor proline-rich tyrosine kinase-2 (PYK2), which is activated by phosphorylation of one or more of its tyrosine residues, has been implicated in the regulation of GLUT4 glucose transporter translocation and glucose transport. Some data favor a positive role of PYK2 in stimulating glucose transport, whereas other studies suggest that PYK2 may participate in the induction of insulin resistance. To ascertain the importance of PYK2 in the setting of obesity and insulin resistance, we (1) evaluated the regulation of PYK2 in mice fed a high fat diet and (2) characterized body and glucose homeostasis in wild type (WT) and PYK2(-/-) mice on different diets. We found that both PYK2 expression and phosphorylation were significantly increased in liver and adipose tissues harvested from high-fat diet fed mice. Wild type and PYK2(-/-) mice were fed a high-fat diet for 8 weeks to induce insulin resistance/obesity. Surprisingly, in response to this diet PYK2(-/-) mice gained significantly more weight than WT mice (18.7+/-1.2g vs. 9.5+/-0.6g). Fasting serum leptin and insulin and blood glucose levels were significantly increased in high-fat diet fed mice irrespective of the presence of PYK2 protein. There was a close correlation between serum leptin and body weight. Intraperitoneal glucose tolerance tests revealed that as expected, the high-fat diet resulted in increased blood glucose levels following glucose administration in wild type mice compared to those fed normal chow. An even greater increase in blood glucose levels was observed in PYK2(-/-) mice compared to wild type mice. These results demonstrate that a lack of PYK2 exacerbates weight gain and development of glucose intolerance/insulin resistance induced by a high-fat diet, suggesting that PYK2 may play a role in slowing the development of obesity, insulin resistance, and/or frank diabetes. PMID- 16039994 TI - Human resistin stimulates the pro-inflammatory cytokines TNF-alpha and IL-12 in macrophages by NF-kappaB-dependent pathway. AB - Resistin, a recently discovered 92 amino acid protein involved in the development of insulin resistance, has been associated with obesity and type 2 diabetes. The elevated serum resistin in human diabetes is often associated with a pro inflammatory milieu. However, the role of resistin in the development of inflammation is not well understood. Addition of recombinant human resistin protein (hResistin) to macrophages (both murine and human) resulted in enhanced secretion of pro-inflammatory cytokines, TNF-alpha and IL-12, similar to that obtained using 5 microg/ml lipopolysaccharide. Both oligomeric and dimeric forms of hResistin were able to activate these cytokines suggesting that the inflammatory action of resistin is independent of its conformation. Heat denatured hResistin abrogated cytokine induction while treatment of recombinant resistin with polymyxin B agarose beads had no effect thereby ruling out the role of endotoxin in the recombinant hResistin mediated cytokine induction. The pro inflammatory nature of hResistin was further evident from the ability of this protein to induce the nuclear translocation of NF-kappaB transcription factor as seen from electrophoretic mobility shift assays. Induction of TNF-alpha in U937 cells by hResistin was markedly reduced in the presence of either dominant negative IkappaBalpha plasmid or PDTC, a pharmacological inhibitor of NF-kappaB. A protein involved in conferring insulin resistance is also a pro-inflammatory molecule that has important implications. PMID- 16039995 TI - Thioredoxin suppresses airway hyperresponsiveness and airway inflammation in asthma. AB - Thioredoxin (TRX) is a 12-kDa redox (reduction/oxidation)-active protein that has a highly conserved site (-Cys-Gly-Pro-Cys-) and scavenges reactive oxygen species. Here we examined whether exogenously administered TRX modulated airway hyperresponsiveness (AHR) and airway inflammation in a mouse asthma model. Increased AHR to inhaled acetylcholine and airway inflammation accompanied by eosinophilia were observed in OVA-sensitized mice. Administration of wild-type but not 32S/35S mutant TRX strongly suppressed AHR and airway inflammation, and upregulated expression of mRNA of several cytokines (e.g., IL-1alpha, IL-1beta, IL-1 receptor antagonist, and IL-18) in the lungs of OVA-sensitized mice. In contrast, TRX treatment at the time of OVA sensitization did not improve AHR or airway inflammation in OVA-sensitized mice. Thus, TRX inhibited the asthmatic response after sensitization, but did not prevent sensitization itself. TRX and redox-active protein may have clinical benefits in patients with asthma. PMID- 16039996 TI - Global analysis of altered gene expression during morphogenesis of Candida albicans in vitro. AB - Candida albicans, a commensal of the gastrointestinal and uro vaginal tract can cause life-threatening infections under conditions of lowered immunity of the host. The changes in the host environment that are sensed by the pathogen which elicit this response have not yet been clearly identified. We report here that co incubation with a macrophage cell line in vitro for extended period of time (16 h) leads to lysis of the macrophage cells. The altered condition in growth medium induces differential gene expression of sets of genes. Specifically genes involved in galactose, protein, and lipid metabolism and stress response undergo concerted changes in their transcript levels. Promoter analysis of differentially expressed genes revealed presence of CPH1 and EFG1 transcription factor binding sites. Based on the gene expression profiles and mutant studies we propose that this morphogenetic response of C. albicans under the conditions used in these experiments is mainly through the pathways controlled by the above two regulators and not through the RIM101-dependent pathway. PMID- 16039997 TI - Agonist Met antibodies define the signalling threshold required for a full mitogenic and invasive program of Kaposi's Sarcoma cells. AB - We previously showed that the Kaposi Sarcoma line KS-IMM express a functional Met tyrosine kinase receptor, which, upon HGF stimulation, activates motogenic, proliferative, and invasive responses. In this study, we investigated the signalling pathways activated by HGF, as well as by Met monoclonal antibodies (Mabs), acting as full or partial agonists. The full agonist Mab mimics HGF in all biological and biochemical aspects. It elicits the whole spectrum of responses, while the partial agonist Mab induces only wound healing. These differences correlated with a more prolonged and sustained tyrosine phosphorylation of the receptor and MAPK evoked by HGF and by the full agonist Mab, relative to the partial agonist Mab. Since Gab1, JNK and PI 3-kinase are activated with same intensity and kinetics by HGF and by the two agonist antibodies, it is concluded that level and duration of MAPK activation by Met receptor are crucial for the induction of a full HGF-dependent mitogenic and invasive program in KS cells. PMID- 16039998 TI - Molecular characterization of cytochrome P450 catalyzing hydroxylation of benzoates from the white-rot fungus Phanerochaete chrysosporium. AB - We cloned full-length cDNA (PcCYP1f) encoding one of the cytochrome P450s in the lignin-degrading basidiomycete Phanerochaete chrysosporium, which showed high homology to P450s in the CYP53 family. PcCYP1f was expressed as an active microsomal protein using the methylotrophic yeast Pichia pastoris expression system. Using the microsomal fraction containing PcCYP1f, a typical P450 CO difference spectrum was obtained with absorption maximum at 448nm. Recombinant PcCYP1f catalyzed the hydroxylation of benzoic acid into 4-hydroxybenzoic acid in the presence of NADPH and P. chrysosporium cytochrome P450 oxidoreductase. In contrast to other CYP53 P450s, this enzyme was shown to catalyze the hydroxylation of 3-hydroxybenzoate into 3,4-dihydroxybenzoate. Furthermore, 2- and 3-methylbenzoate were also shown to be substrates of PcCYP1f. This is the first report showing the expression of a functionally active Phanerochaete P450. Finally, real-time quantitative PCR analysis revealed that PcCYP1f is induced at a transcriptional level by exogenous addition of benzoic acid. PMID- 16039999 TI - Aspirin reduces endothelial cell senescence. AB - We report here the effect of aspirin on the onset of replicative senescence. Endothelial cells that were cultured until cumulative population doublings 40 showed clear signs of aging. Incubation with aspirin inhibited senescence associated beta-galactosidase activity and increased telomerase activity. Along with the delayed onset of senescence, aspirin decreased reactive oxygen species and increased nitric oxide (NO) and cGMP levels. Furthermore, aspirin reduced the elaboration of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of NO synthase, and up-regulated the activity of dimethylarginine dimethylaminohydrolase, the enzyme that degrades ADMA. These effects were specific in that other nonsteroidal anti-inflammatory drugs, such as ibuprofen or acetaminophen, did not prevent the onset of endothelial senescence. The NO synthase inhibitor l-NAME, but not its inactive d-enantiomer, led to complete inhibition of aspirin-delayed senescence. These findings demonstrate that aspirin delays the onset of endothelial senescence by preventing a decrease in NO formation/generation. This might provide a therapeutic strategy aimed at blocking aging-induced NO inhibition. PMID- 16040000 TI - Protein fragment complementation in M.HhaI DNA methyltransferase. AB - The 5mC DNA methyltransferase M.HhaI can be split into two individually inactive N- and C-terminal fragments that together can form an active enzyme in vivo capable of efficiently methylating DNA. This active fragment pair was identified by creating libraries of M.HhaI gene fragment pairs and then selecting for the pairs that code for an active 5mC methyltransferase. The site of bisection for successful protein fragment complementation in M.HhaI was in the variable region near the target recognition domain between motif VIII and TRD. This same region is the location of bifurcation in the naturally split 5mC methyltransferase M.AquI, the location for circular permutation in M.BssHII, and the location for previously engineered split versions of M.BspRI. PMID- 16040001 TI - Regulation of zymogen granule exocytosis by Ca2+, cAMP, and PKC in pancreatic acinar cells. AB - The effect of cAMP and PKC on zymogen granule exocytosis was investigated by simultaneously measuring cytosolic Ca2+ concentration ([Ca2+]c) and individual zymogen granule exocytosis in isolated mouse pancreatic acini. When acinar cells were stimulated with acetylcholine (ACh, 10 microM), exocytic events were detected through granule-attached apical membranes with [Ca2+]c rise. Application of secretin, forskolin (an adenylate cyclase activator), or PMA (a PKC activator) alone did not elicit any [Ca2+]c rise or zymogen granule exocytosis, but co stimulation with ACh led to exocytosis in that the total number of secreted granules increased markedly without a significant difference in [Ca2+]c rises. When we evoked exocytosis by [Ca2+]c ramps, pretreatment with forskolin or PMA elicited exocytosis at lower [Ca2+]c levels. These results indicate that PKC or cAMP alone could not directly elicit zymogen granule exocytosis, but that they increase the total releasable pool by rendering zymogen granules more sensitive to Ca2+. PMID- 16040002 TI - The role of the central L- or D-Pro residue on structure and mode of action of a cell-selective alpha-helical IsCT-derived antimicrobial peptide. AB - IsCT-P (ILKKIWKPIKKLF-NH2) is a novel alpha-helical antimicrobial peptide with bacterial cell selectivity designed from a scorpion-derived peptide IsCT. To investigate the role of L- or D-Pro kink on the structure and the mode of action of a short alpha-helical antimicrobial peptide with bacterial cell selectivity, we synthesized IsCT-p, in which D-Pro is substituted for L-Pro8 of IsCT-P. CD spectra revealed that IsCT-P adopted a typical alpha-helical structure in various membrane-mimicking conditions, whereas IsCT-p showed a random structure. This result indicated that D-Pro in the central position of a short alpha-helical peptide provides more remarkable structural flexibility than L-Pro. Despite its higher antibacterial activity, IsCT-p was much less effective at inducing dye leakage in the negatively charged liposome mimicking bacterial membrane and induced no or little membrane potential depolarization of Staphylococcus aureus. Confocal laser scanning microscopy showed that IsCT-p penetrated the bacterial cell membrane and accumulated in the cytoplasm, whereas IsCT-P remained outside or on the cell membrane. These results suggested that the major target of IsCT-P and IsCT-p is the bacterial membranes and intracellular components, respectively. Collectively, our results demonstrated that the central D-Pro kink in alpha helical antimicrobial peptides plays an important role in penetrating bacterial membrane as well as bacterial cell selectivity. PMID- 16040003 TI - RNAi-mediated gene silencing to assess the role of synaptobrevin and cystatin in tick blood feeding. AB - In addition to being the conduit for pathogens into hosts, tick saliva contains a broad array of secretory products that facilitate prolonged tick attachment and blood feeding. Proteins found in tick saliva modulate host hemostasis and immune responses. However, it is not clear whether ticks manipulate the immune responses of their hosts by disrupting the antigen-processing pathways of the hosts. Protein secretion into tick saliva from the salivary glands is due to exocytosis of vesicular membrane-bound granular material regulated by SNARE complex proteins. Proteins associated with vesicles (v-SNAREs) are essential components of the exocytotic process. In this study, we assessed the functional significance of synaptobrevin, a SNARE protein, and cystatin, a cysteine protease inhibitor to blood feeding success, in the lone star tick, Amblyomma americanum, using in vivo RNA interference. In separate experiments, tick salivary cystatin and synpatobrevin genes were silenced by injecting adult ticks with 500 ng of dsRNA complementing each gene sequence. Silencing was demonstrated by reduced transcript in midguts and salivary glands. Additionally, disrupting expression of cystatin and synaptobrevin by RNAi reduced the ability of ticks to feed successfully, as demonstrated by feeding inhibition and reduced engorgement weights. Moreover, normal ticks exposed to a rabbit previously exposed to cystatin-silenced ticks exhibited significant resistance to tick feeding. Based on these findings, ticks appear to skillfully evade the host immune system by secreting cystatin, which disrupts normal antigen processing in antigen presenting cells of hosts. PMID- 16040004 TI - Muscarinic receptors are involved in LMM3 tumor cells proliferation and angiogenesis. AB - Angiogenesis is a process of new blood vessel development from pre-existing vasculature and it plays an essential role in tumor growth and metastases. Here, we investigate the expression of muscarinic acetylcholine receptors (mAchR) and their participation in tumor cell proliferation and angiogenesis ability. Saturation binding assays with the tritiated muscarinic antagonist quinuclidinyl benzilate indicate that LMM3 cells derived from a murine mammary adenocarcinoma express a single class of functional mAchR. Competition binding assays with selective muscarinic antagonists indicate a predominance of M3 receptor subtype. The muscarinic agonist carbachol (CARB) stimulates LMM3 cell proliferation in a concentration dependent manner. The maximal effect induced by 10(-9)M CARB was totally blunted by atropine and by the selective M3 and M1 antagonists, para fluoro hexahydro sila-difenidol (pf-HHSiD) and pirenzepine, respectively. In addition, pf-HHSiD completely blocked in vivo CARB-induced neovascular formation and vascular endothelial growth factor-A in LMM3 tumor cells. We can conclude that mAchR expressed in LMM3 mammary tumor cells positively regulate proliferation and angiogenesis required for tumor progression. PMID- 16040005 TI - Cerebral asymmetry and behavioral lateralization in rats chronically lacking n-3 polyunsaturated fatty acids. AB - BACKGROUND: Anatomic and functional brain lateralization underlies hemisphere specialization for cognitive and motor control, and deviations from the normal patterns of asymmetry appear to be related to behavioral deficits. Studies on n-3 polyunsaturated fatty acid (PUFA) deficiency and behavioral impairments led us to postulate that a chronic lack of n-3 PUFA can lead to changes in lateralized behavior by affecting structural or neurochemical patterns of asymmetry in motor related brain structures. METHODS: We compared the effects of a chronic n-3 PUFA deficient diet with a balanced diet on membrane phospholipid fatty acids composition and immunolabeling of choline acetyltransferase (ChAt), as a marker of cholinergic neurons, in left and right striatum of rats. Lateral motor behavior was assessed by rotation and paw preference. RESULTS: Control rats had an asymmetric PUFA distribution with a right behavioral preference, whereas ChAt density was symmetrical. In deficient rats, the cholinergic neuron density was 30% lower on the right side, associated with a loss of PUFA asymmetry and behavior laterality. They present higher rotation behavior, and significantly more of them failed the handedness test. CONCLUSION: These results indicate that a lack of n-3 PUFA is linked with a lateral behavior deficit, possibly leading to cognitive disturbances. PMID- 16040006 TI - Association study of the A2M and LRP1 Genes with Alzheimer disease in the Han Chinese. AB - BACKGROUND: Low-density lipoprotein receptor-related protein 1 (LRP1) and alpha-2 macroglobulin (A2M) are two plausible candidate genes for Alzheimer disease (AD) based on their important biological function and positional information. To date, numerous studies have investigated their possible association with AD but the results are controversial. METHODS: To investigate the potential genetic contribution of the two genes in the Han Chinese population, we performed a case control association study using 10 polymorphisms (4 in LRP1 and 6 in A2M) that span approximately the whole corresponding gene. RESULTS: Comparison of allele, genotype, and haplotype frequencies for polymorphisms in A2M revealed no significant differences between patients and control subjects. For the LRP1 gene, however, we found an overrepresentation of the CTCG haplotype in the control group (p = .002). The difference was still of statistical significance in the apolipoprotein E (APOE) epsilon 4 negative subjects (p(CTCG) = .003). Multiple logistic regression analysis did not show any evidence of synergism between A2M, LRP1, and APOE. CONCLUSIONS: Our results indicate that the CTCG haplotype of LRP1 may reduce the risk of late-onset AD, but A2M is not associated with this disease in the Han Chinese population. PMID- 16040007 TI - Nutrition and depression: implications for improving mental health among childbearing-aged women. AB - Adequate nutrition is needed for countless aspects of brain functioning. Poor diet quality, ubiquitous in the United States, may be a modifiable risk factor for depression. The objective was to review and synthesize the current knowledge of the role of nutrition in depression, and address implications for childbearing aged women. Poor omega-3 fatty acid status increases the risk of depression. Fish oil and folic acid supplements each have been used to treat depression successfully. Folate deficiency reduces the response to antidepressants. Deficiencies of folate, vitamin B12, iron, zinc, and selenium tend to be more common among depressed than nondepressed persons. Dietary antioxidants have not been studied rigorously in relation to depression. Childbearing-aged women are particularly vulnerable to the adverse effects of poor nutrition on mood because pregnancy and lactation are major nutritional stressors to the body. The depletion of nutrient reserves throughout pregnancy and a lack of recovery postpartum may increase a woman's risk of depression. Prospective research studies are needed to clarify the role of nutrition in the pathophysiology of depression among childbearing-aged women. Greater attention to nutritional factors in mental health is warranted given that nutrition interventions can be inexpensive, safe, easy to administer, and generally acceptable to patients. PMID- 16040008 TI - The 5-HT2C receptor antagonist RO 60-0491 counteracts rats' retention deficits in a recognition memory task. AB - The effects of the 5-HT2C receptor antagonist RO 60-0491 on recognition memory were investigated in the rat by using the object recognition task. Post-training administration of RO 60-0491 at 3 mg/kg, but not at 1 mg/kg, counteracted rats' performance deficits in the object recognition task, suggesting that this 5-HT2C receptor antagonist compound modulates storage and/or retrieval of information. PMID- 16040009 TI - Brn-3a is required for the generation of proprioceptors in the mesencephalic trigeminal tract nucleus. AB - The distribution of motor and proprioceptive neurons was investigated in the trigeminal nervous system of wild-type and Brn-3a knockout mice at embryonic day 18.5 and postnatal day 0. We found that the trigeminal motor nucleus (Mo5) contained abundant motoneurons in wild-type (mean number +/- SD per section = 128 +/- 22, range = 93-167) and knockout (mean number +/- SD per section = 121 +/- 23, range = 75-158) mice and that the cell size of Mo5 neurons was similar between these mice (wild-type, mean +/- SD = 165 +/- 59 microm2, range = 65-326 microm2; knockout, mean +/- SD = 167 +/- 59 microm2, range = 71-327 microm2). Mo5 neurons were immunoreactive for calcitonin gene-related peptide and such immunoreactive neurons were abundant in both wild-type and mutant mice. In the mesencephalic tract nucleus (Mes5) of wild-type mice, many proprioceptors (mean number +/- SD per section = 56 +/- 19, range = 27-85) that contained parvalbumin immunoreactivity were also observed. In knockout mice, however, Mes5 neurons could not be detected. The area of brainstems which normally contained the Mes5 was devoid of parvalbumin-immunoreactive proprioceptors. The present study suggests that Brn-3a is required for the development of proprioceptors but not motoneurons in the trigeminal nervous system. PMID- 16040010 TI - Glutamate induces the expression and release of tumor necrosis factor-alpha in cultured hypothalamic cells. AB - Tumor necrosis factor-alpha (TNFalpha) affects several CNS functions such as regulation of sleep, body temperature, and feeding during pathology. There is also evidence for TNFalpha involvement in physiological sleep regulation, e.g., TNFalpha induces sleep and brain levels of TNFalpha increase during prolonged wakefulness. The immediate cause of enhanced TNFalpha production in brain is unknown. We investigated whether glutamate could signal TNFalpha production because glutamate is a neurotransmitter associated with cell activation and wakefulness. We used primary cultures of fetal rat hypothalamic cells to examine the expression and release of TNFalpha. Immunostaining for neuron specific enolase revealed that the cultures were 50-60% neuronal and 40-50% non-neuronal cells. TNFalpha was detected in both the media and cells under basal conditions. Stimulation of the cells with 1 mM glutamate for 2 h produced an increase in media content of TNFalpha, whereas cell content was elevated at earlier time points. Using trypan blue exclusion and MTT assays, there was no evidence of cell toxicity with this stimulation protocol. Immunocytochemical staining revealed that TNFalpha was expressed by approximately 25% of the neurons and approximately 75% of the glial cell in the culture. Stimulation of the cultures with glutamate did not increase the percentage of cells expressing TNFalpha. We conclude that TNFalpha is constitutively expressed and released by healthy cultures of hypothalamic cells and that activation of the cells with a non-toxic challenge of glutamate increases TNFalpha production. These findings support the hypothesis that TNFalpha can participate in normal physiological regulation of sleep and feeding. PMID- 16040012 TI - A comparison of serial halving and the rule of nines as a pre-hospital assessment tool in burns. AB - Following endorsement of serial halving by the Faculty of Pre-Hospital Care of the Royal College of Surgeons of Edinburgh this study aimed to determine whether the technique was comparable to the rule of nines in making initial assessments of body surface area burned. Ten 'casualties' were made up to represent burn victims (range 6-61%). An external panel of six consultants and one specialist registrar in plastic surgery were invited to assess the simulated casualties. They gave individual and a consensus estimate of the burned areas. One hundred and twenty-five members of local emergency services and military paramedical staff were given a brief video and slide presentation describing either the rule of nines or serial halving method of burn area assessment. These techniques were then used to assess the 10 simulated casualties, giving 1250 estimates of burn surface area. The understanding of both techniques appeared adequate in both test groups. Estimates from serial halving and rule of nines groups differed from the assessments of the external panel. No statistical difference was demonstrated between serial halving and the rule of nines as an initial assessment tool when determining disposal. Serial halving has an inherent weakness when assessing certain sizes of burn. The rule of nines requires that the assessor knows and understands the proportionate areas of the body. The mathematics of percentages and fractions appeared to confuse some assessors. PMID- 16040011 TI - Murine brain capillary endothelial cells exhibit improved barrier properties under the influence of hydrocortisone. AB - Hydrocortisone is known to induce barrier properties in porcine primary cultures of microvascular endothelial cells. Here we present similar effects of hydrocortisone on a serum-free in vitro model based on primary cultured mouse brain endothelial cells. These cells in culture express typical blood-brain barrier properties and the transendothelial electrical resistance is enhanced after the addition of hydrocortisone to the medium in physiological concentrations. The improvement of the barrier is accompanied by changes at the cell borders indicated by immunofluorescence staining of tight junction proteins. Transmission electron microscopy imaging indicates morphological changes at the cell-cell contact zones which correlates to the observed changes in the transendothelial electrical resistance after HC supplementation. Phalloidin staining of F-actin shows a rearrangement to "fiber-like" structures in the longitudinal direction of the cell. These findings together with additional electrical impedance analysis of the monolayer suggest that several changes including cell-cell contact alteration, cell-substrate attachment and cytoskeletal rearrangements cause enhanced barrier properties in this murine endothelial culture. The present data are consistent with earlier findings in a porcine serum-free in vitro model. Thus, evidence is given that the barrier enforcement induced by glucocorticoids is not a species-specific effect and that the barrier improvement is correlated with a change of the cell morphology rather than changes in tight junction protein expression. PMID- 16040013 TI - The nodal neck level of sentinel lymph nodes in mucosal head and neck cancer. AB - Sentinel node biopsy is emerging as a successful means of identifying subclinical lymph node disease in mucosal head and neck cancer. Sentinel node studies in melanoma and breast cancer have identified sentinel nodes at unusual sites and the technique is redefining our understanding of dynamic lymphatic flow. In this study, the sentinel nodes in mucosal head and neck malignancies were mapped according to their site within the neck and this was correlated with tumour site within the oral cavity. Fifty-two necks were explored for sentinel nodes from tumours located in the tongue (23 cases), floor of mouth (12 cases), palate (six cases), retromolar trigone (five cases), alveolus (three cases), buccal mucosa (two cases), tonsil (two cases) and lip (one case). In total, 124 sentinel nodes were found in levels I-V. Two hot spots were found in the tonsils and were not excised, two nodes were located in level IIB, four nodes were found in level IV, three in the contralateral neck and one in level V. The sentinel nodes located at unusual sites would not have been excised in a supraomohyoid neck dissection and the study has improved our understanding of dynamic lymph flow from tumours. PMID- 16040014 TI - Motorneuron protection by N-acetyl-cysteine after ventral root avulsion and ventral rhizotomy. AB - Motor recovery after proximal nerve injury remains extremely poor, despite advances in surgical care. Several neurobiological hurdles are implicated, the most fundamental being extensive cell death within the motorneuron pool. N-acetyl cysteine almost completely protects sensory neurons after peripheral axotomy, hence its efficacy in protecting motorneurons after ventral root avulsion/rhizotomy was investigated. In adult rats, the motorneurons supplying medial gastrocnemius were unilaterally pre-labelled with retrograde tracer (true blue/fluoro-gold), prior to L5 and 6 ventral root avulsion, or rhizotomy. Groups received either intraperitoneal N-acetyl-cysteine (ip, 150 or 750 mg/kg/day), immediate or delayed intrathecal N-acetyl-cysteine treatment (it, 2.4 mg/day), or saline; untreated animals served as controls. Either 4 (avulsion model) or 8 (rhizotomy model) weeks later, the pre-labelled motorneurons' mean soma area and survival were quantified. Untreated controls possessed markedly fewer motorneurons than normal due to cell death (avulsion 53% death; rhizotomy 26% death, P<0.01 vs. normal). Motorneurons were significantly protected by N-acetyl cysteine after avulsion (ip 150 mg/kg/day 40% death; it 30% death, P<0.01 vs. no treatment), but particularly after rhizotomy (ip 150 mg/kg/day 17% death; ip 750 mg/kg/day 7% death; it 5% death, P<0.05 vs. no treatment). Delaying intrathecal treatment for 1 week after avulsion did not impair neuroprotection, but a 2-week delay was deleterious (42% death, P<0.05 vs. 1-week delay, 32% death). Treatment prevented the decrease in soma area usually found after both types of injury. N acetyl-cysteine has considerable clinical potential for adjuvant treatment of major proximal nerve injuries, including brachial plexus injury, in order that motorneurons may survive until surgical repair facilitates regeneration. PMID- 16040015 TI - Measurement of perfusion pressure of perforators and its correlation with their internal diameter. PMID- 16040016 TI - Islanded tripier flap--another useful variant. PMID- 16040017 TI - Internal mammary vessels, recipient vessels of choice for free tissue breast reconstruction? PMID- 16040018 TI - Conflicts in the treatment of chronic ulcers in drug addicts--case series and discussion. AB - Vascular access in intravenous drug abusers may become compromised by the repeated injection of toxic substances. In such circumstances abusers are driven by their addiction to seek alternative routes of drug delivery. We report a series of individuals with chronic ulcers, which were cultivated and maintained for the administration of heroin. We advise that practitioners should be wary of granulation tissue being promoted in this way and suggest that the patients' desire for wound healing may be overridden by their addiction and that successful treatment of these wounds is reliant upon cessation of drug abuse and patient compliance. In our experience this is most easily achieved in a multidisciplinary environment. PMID- 16040019 TI - The cell based dressing with living allogenic keratinocytes in the treatment of foot ulcers: a case study. AB - In this study, we aimed to investigate the efficacy of cell based dressing with living allogenic keratinocytes in diabetic foot patients. To address this issue, the cultured keratinocytes were attached to the microcarriers produced from polyethylene and silica. The microcarriers were then applied to the wounds at 3 day intervals. Forty patients with grade II and III diabetic foot ulcers were included into the study. The patients were randomised into two groups (n=20). The treatment and control groups received cell based dressing and microcarriers kept in culture medium overnight, respectively. The wound size was recorded at 3 days intervals. The wounds were also categorised by a specific scoring system considering the wound contraction, granulation tissue formation, epithelisation and discharge from the wounds. The high score indicates better condition. The mean reduction of the wound area was 92% in the treatment group and 32% in the control group at the end of the 30 days treatment (p<0.001). When considered the complete healing, the mean number of dressings was 9.2+/-3.2 in the treatment group whereas it was 16.5+/-2.3 in the control group (p<0.001). The initial mean score of the treatment and control groups were 2.5 and 2.35, respectively. At the end of the 30th day, the mean score of the treatment group was 17.15+/-2.7 and of control group was 9.05+/-3. Allogenic keratinocyte treatment delivered with microcarriers can make significant contributions to wound healing in diabetic foot patients. PMID- 16040020 TI - Ultrasound-accelerated enzymatic synthesis of sugar esters in nonaqueous solvents. AB - Comparative studies of enzymatic synthesis of glucose esters under ultrasound and shaking were carried out in nonaqueous media. The influence of solvents, enzymes, chain length of the acyl donors, the power of the ultrasound bath, and intermittent ultrasound on the enzymatic synthesis was investigated. Among the eight solvents selected, pyridine was the most appropriate with alkaline protease from Bacillus subtilis whether under ultrasound or shaking. The acceleration effect of ultrasound with Novozym 435 and the alkaline protease from B. subtilis catalyzed transesterification increased with the chain length of acyl donors, decreasing from C(10) to C(4). We also investigated the influence of the power (50, 100, and 120 W) of the ultrasound irradiation and the manner of operation (continuous ultrasound, 10 min ultrasound/20 min shaking without ultrasound) on the transesterification. The results showed that higher power and continual operational gave the better acceleration. Ultrasound did not change the character and selectivity of the enzyme in the transesterification. PMID- 16040021 TI - Salt-assisted acid hydrolysis of chitosan to oligomers under microwave irradiation. AB - The effect of inorganic salts such as sodium chloride on the hydrolysis of chitosan in a microwave field was investigated. While it is known that microwave heating is a convenient way to obtain a wide range of products of different molecular weights only by changing the reaction time and/or the radiation power, the addition of some inorganic salts was shown to effectively accelerate the degradation of chitosan under microwave irradiation. The molecular weight of the degraded chitosan obtained by microwave irradiation was considerably lower than that obtained by traditional heating. Moreover, the molecular weight of degraded chitosan obtained by microwave irradiation assisted under the conditions of added salt was considerably lower than that obtained by microwave irradiation without added salt. Furthermore, the effect of ionic strength of the added salts was not linked with the change of molecular weight. FTIR spectral analyses demonstrated that a significantly shorter time was required to obtain a satisfactory molecular weight by the microwave irradiation-assisted inorganic salt method than by microwave irradiation without inorganic salts and conventional technology. PMID- 16040022 TI - Endothelin-1 and isoprenaline co-stimulation causes contractile failure which is partially reversed by MEK inhibition. AB - OBJECTIVE: The mitogen-activated kinase kinases (MEK)-extracellular signal regulated kinases (ERK) signaling pathway is activated by agonists like catecholamines or endothelin-1 (ET-1) and has been implicated in cardiac pathology, such as the progression from cardiac hypertrophy to failure. The purpose of the present study, performed in an in vitro model of contractile failure, was to evaluate whether MEK inhibition prevents functional deterioration. METHODS AND RESULTS: Contractile dysfunction was induced in reconstituted rat heart tissue by concomitant treatment with ET-1 (10 nmol/l) and isoprenaline (ISO, 10 nmol/l) for 5 days. While basal force of contraction was unchanged, contractile responsiveness to beta-adrenoceptor agonists was markedly impaired (active force declined to 51% of controls) and was associated with decreased lusitropy. Moreover, in ET-1+ISO-treated heart tissues, reprogramming of gene expression was observed with an increased ratio of beta-myosin heavy chain (MHC) to alpha-MHC mRNA and increased transcript levels of ANF and skeletal/smooth muscle alpha-actin isoforms. The MEK inhibitor U0126 (10 micromol/l) almost completely prevented the reduction in beta-adrenergic responsiveness and the negative lusitropic effect of ET-1+ISO co-stimulation. In addition, U0126 completely normalized ANF gene expression, but did not affect or only marginally affected expression of MHC and alpha-actin isoforms. CONCLUSIONS: These results suggest that interruption of the MEK-ERK signaling pathway with a specific MEK inhibitor prevents, in part, the occurrence of a pathologic phenotype secondary to excessive stimulation with neurohumoral factors. The MEK ERK pathway seems to be an important but not exclusive regulatory pathway responsible for the development of contractile dysfunction. PMID- 16040023 TI - Increased mortality associated with TCDD exposure in mice infected with influenza A virus is not due to severity of lung injury or alterations in Clara cell protein content. AB - Most studies examining the cause of increased mortality in mice infected with a normally non-lethal dose of influenza A virus after exposure to 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) have focused on defects in the immune system. This study examined other possible consequences of TCDD exposure, which could alter pulmonary inflammation during infection. We measured bronchoalveolar lavage (BAL) fluid lactate dehydrogenase (LDH) and protein concentrations and lung wet to dry weight ratios to assess lung damage and edema formation. Immunohistochemistry for Cyp1A1 was used to evaluate the responsiveness of the lung to TCDD. Additionally, we characterized the effects of TCDD on Clara cell secretory protein (CCSP), which plays a regulatory role in pulmonary inflammation. There were no differences in BAL fluid LDH and protein levels, lung wet to dry weight ratios, or the amount of CCSP in the lungs from mice treated with TCDD or vehicle control. The amount of Cyp1A1 in endothelial cells, Clara cells, and Type II pneumocytes was greatly induced after TCDD exposure. Although lung tissue was clearly responsive to TCDD as shown by Cyp1A1 induction, the increased mortality in infected mice exposed to TCDD did not correlate with increased damage to the lung or decreased CCSP concentrations. PMID- 16040024 TI - Comparative aspects of sperm membrane fatty acid composition in silver (Vulpes vulpes) and blue (Alopex lagopus) foxes, and their relationship to cell cryopreservation. AB - Cryogenic protocols have been developed for the storage of farmed silver fox (Vulpes vulpes) spermatozoa. However, these same protocols and modifications of these protocols have failed to satisfactorily preserve spermatozoa collected from farmed blue foxes (Alopex lagopus). Because cryogenic success has been linked to membrane composition, the plasma membrane lipid composition of farmed blue fox and silver fox spermatozoa was studied. Silver fox spermatozoal membranes have significantly higher levels of docosapentaenoic acid (DPA; 22:5, n-6) compared to blue fox spermatozoa, and blue fox spermatozoal membranes have significantly higher levels of stearic acid (18:0). Silver fox spermatozoal membranes not only have a higher ratio of unsaturated/saturated membrane fatty acids, but also higher levels of membrane desmosterol and cholesterol. PMID- 16040025 TI - The role of combinational coding by homeodomain and bHLH transcription factors in retinal cell fate specification. AB - Two major families of transcription factors (TFs), basic helix-loop-helix (bHLH) and homeodomain (HD), are known to be involved in cell fate identity. Some recent findings suggest that these TFs are used combinatorially to code for cellular determination in the retina. However, neither the extent nor the efficiency of such a combinatorial coding mechanism has been tested. To look systematically for interactions between these two TF types that would address these questions, we used a matrix analysis. We co-expressed each of six retinally expressed bHLH TFs (XNeuroD; XNgnr-1; Xath3; Xath5; Xash1; Xash3) with each of eight retinally expressed HD TFs (XRx1; XOptx2; XSix3; XPax6; XOtx2; XOtx5b; XBH; XChx10) in retinal progenitors of Xenopus laevis using targeted lipofection. The effects of each of these combinations were assayed on the six major cell types in the retina: Retinal ganglion cells (GCs), Amacrines (ACs), Bipolars (BCs), Horizontals (HCs), Photoreceptors (PRs), and Muller cells (MCs), creating 288 result categories. Multiple-way ANOVA indicated that in 14 categories, there were interactions between the two TFs that produced significantly more or less of a particular cell type than either of the components alone. However, even the most effective combinations were incapable of generating more than 65% of any particular cell type. We therefore used the same techniques to misexpress selected combinations of three TFs in retinal progenitors, but found no further enhancements of particular cell fates, indicating that other factors are probably involved in cell type specification. To test whether particular combinations were essential for horizontal fates, we made VP16 and EnR fusion constructs of some of the factors to provide dominant negative transcriptional activities. Our results confirmed that normal activities of certain combinations were sufficient, and that individually these activities were important for this fate. PMID- 16040026 TI - Microarray analysis of localization of maternal transcripts in eggs and early embryos of the ascidian, Ciona intestinalis. AB - The establishment of body axes and specification of early embryonic cells depend on maternally supplied transcripts and/or proteins, several of which are localized at specific regions of fertilized eggs and early embryos. The ascidian is known to exhibit a mosaic mode of development, and this mode is largely dependent on localized maternal factors. Using blastomere isolation, microarray and whole-mount in situ hybridization, the present study of Ciona intestinalis demonstrates that maternal transcripts of a total of 17 genes are localized at the posterior-most region of fertilized eggs and early embryos. Ten of them are newly identified in the present study, while the remaining seven genes have already been characterized in previous studies. In addition, maternal transcripts of two genes, in addition to 14 genes encoded by the mitochondrial genome, showed a mitochondria-like distribution. Despite the present comprehensive approach, we could not identify maternal transcripts that are clearly localized to the animal pole side, the vegetal-pole side, the anterior-side or other specific regions of the early embryo. Therefore, we concluded that the posterior-most localization and mitochondria-like distribution appear to be major specialized patterns of maternal transcripts in early Ciona embryos. PMID- 16040027 TI - Imaging in situ protein-DNA interactions in the cell nucleus using FRET-FLIM. AB - Although the distribution of DNA-binding proteins inside the cell nucleus can be analyzed by immunolabeling or by tagging proteins with GFP, we cannot establish whether the protein is bound to DNA or not. Here, we describe a novel approach that allows imaging of the in situ interaction between a GFP-fusion protein and DNA in the cell nucleus, using fluorescence resonance energy transfer (FRET). We used fluorescence lifetime imaging microscopy (FLIM) as a reliable tool to detect protein in contact with DNA. The method was successfully applied to the DNA binding proteins histone H2B and the glucocorticoid receptor and to the heterochromatin-associated proteins HP1alpha and HP1beta. PMID- 16040028 TI - Proteoglycans in axonal regeneration. PMID- 16040029 TI - Synthesis and biological evaluation of new thiazolyl/benzothiazolyl-amides, derivatives of 4-phenyl-piperazine. AB - A series of thiazolyl-N-phenyl piperazines has been synthesised and tested for anti-inflammatory activity. Their R(M) values were determined as an expression of their lipophilicity. Theoretical calculation of their lipophilicity, as clog P and logPsk also performed. The effect of the synthesised compounds on inflammation, using the carrageenin induced mouse paw oedema model was studied. In general, the studied compounds were found to be potent anti-inflammatory agents (44-74.1%). Anti-inflammatory activity was influenced by some structural characteristics of the synthesised compounds. An attempt was made to correlate their biological activity with some physicochemical parameters using a quantitative structure-activity relationship approach (QSAR). PMID- 16040030 TI - Synthesis of 2-azetidinones from 2-diazo-1, 2-diarylethanones and N-(2 thienylidene)imines as possible antimicrobial agents. AB - An equimolar reaction of 2-diazo-1, 2-diarylethanones with N-(2 thienylidene)imines affords 1-substituted-3, 3-diaryl-4-(2-thienyl)-2 azetidinones in excellent yields. The products have been characterized on the basis of satisfactory analytical and spectral (IR, 1H and 13C NMR, MS) data. The mechanism of formation of the products is shown. The antimicrobial activity of the compounds against some Gram(+) and Gram(-) bacteria, and fungi is reported. PMID- 16040031 TI - PEG-metronidazole conjugates: synthesis, in vitro and in vivo properties. AB - Metronidazole (MTZ), a drug used for the treatment of protozoal infections caused by protozoa and anaerobic microorganisms, was conjugated to linear or branched poly(ethylene glycol) of 5,000, 10,000 and 20,000 Da. An ester linkage between polymer and drug was used in the coupling to yield a polymeric prodrug. The modification allowed overcoming the known MTZ solubility problem leading us to obtain a bioconjugate more suitable for parental administration. The conjugates of various molecular weight polymers have been tested in vitro toward chemical degradation and digestive enzymes. It was found that molecular weight and shape of PEG is critical for the prodrugs stability. Good resistance in the stomach acidic media was found and a slow release of the drug in the large intestinal fluid may take place. In vivo studies carried out following i.v. or s.c. administration to mice revealed improved pharmacokinetics properties upon conjugation. PMID- 16040032 TI - Thyroid hormones inhibit frog corticotropin-releasing factor-induced thyrotropin release from the bullfrog pituitary in vitro. AB - Due to the lack of a radioimmunoassay (RIA) system for amphibian thyrotropin (TSH), no direct evidence that thyroid hormone suppresses the release of TSH from the amphibian pituitary has been obtained. However, we recently developed an RIA for bullfrog (Rana catesbeiana) TSH and thus were able to study the effect of thyroid hormone on the release of TSH from the bullfrog pituitary. Enzymatically dispersed pituitary cells of larval, juvenile, and adult bullfrogs were cultured in the absence or presence of 100 nM corticotropin-releasing factor of bullfrog origin (fCRF), which is known to be a potent stimulator of the release of TSH. The amount of spontaneously released TSH was higher in late prometamorphic and climactic tadpoles than in early prometamorphic larvae and juvenile and adult frogs. Pituitary cells from tadpoles at metamorphic climax responded to fCRF to release much more TSH than those from early and late prometamorphic tadpoles and juvenile and adult frogs. In all cases, the fCRF (100 nM)-induced, but not the basal, release of TSH was significantly suppressed by 1 nM triiodothyronine (T(3)) and 1000 nM thyroxine (T(4)), when examined using adult pituitary cells. The suppressive effect of thyroid hormones was revealed to be dependent on their concentrations. PMID- 16040033 TI - A longitudinal study of digit ratio (2D:4D) and other finger ratios in Jamaican children. AB - It has been hypothesised that the ratio between the length of the 2nd and 4th digits (2D:4D) is a correlate of prenatal sex steroids, and this relationship is strongest for the right hand. Furthermore, it has been suggested that 2D:4D is sexually dimorphic, the dimorphism is determined early, and 2D:4D among children is stable with growth. Here, we present the first longitudinal study of right and left hand 2D:4D. Our sample was 108 (54 males) Jamaican children. The first measurements were made in 1998 when mean age was 9.68 +/- 1.39 years, and a second set of measurements were made in 2002. We found that: (i) there was a small increase in 2D:4D with age which was lowest in the right hand; (ii) 2D:4D was sexually dimorphic, the means for males and females differed in the same direction in the 1998 and 2002 samples, and the sex difference was significant in the 1998 but not in the 2002 sample; (iii) the correlation between the 1998 and 2002 measurements of 2D:4D was high, indicating that rank order of the ratio was stable across year groups; and (iv) the rate of change in 2D:4D did not differ significantly across year groups. We conclude that 2D:4D increases slightly with age in children with the effect less marked for the right hand (i.e. the hand which is likely to show the strongest association with prenatal steroids), 2D:4D is sexually dimorphic from an early age, and the rank order of 2D:4D is stable in children. We discuss the implications of our findings for the status of 2D:4D as a correlate of prenatal sex steroids. The patterns of change in other finger ratios are also considered. PMID- 16040034 TI - Hydrostatic reduction of acute uterine inversion. PMID- 16040035 TI - Abdominal wall endometriomas. AB - OBJECTIVE: To investigate the clinical characteristics, treatment, and factors of recurrence of abdominal wall endometriomas (AWE). METHOD: Sixty-four cases of AWE diagnosed at Peking Union Medical College Hospital (PUMCH) from 1983 to 2003 were reviewed retrospectively. RESULT: There was an AWE incidence of 0.044% among the parturients undergoing cesarean section at PUMCH, of whom 87.5% had the typical complaint of an enlarging mass and pain during menstruation. Among these women, 62 underwent low abdominal surgery for endometrioma (2 for primary umbilicus endometrioma); 2 women with small endometriomas opted for a temporary medical solution and had relief after menopause. The latent period of AWE positively correlated to the women's age at onset of symptoms (P<0.001). Of the 62 women who underwent local excision, 19 had an unsatisfactory experience with medical management. There were 5 recurrences and 1 evolution to malignancy during a mean follow-up of 83.7 months. Recurrence was closely related to the size and depth of lesions. CONCLUSION: Because of its typical clinical manifestations, abdominal wall endometriomas could be diagnosed before pregnancy. Surgical excision is the only effective treatment and wide local excision with clear margins is the key point to prevent recurrence. PMID- 16040036 TI - Adsorption treatment and recovery of the hazardous dye, Brilliant Blue FCF, over bottom ash and de-oiled soya. AB - Two waste materials-bottom ash, a power plant waste, and de-oiled soya, an agricultural waste-are meticulously and successfully used as adsorbent for the removal and recovery of a hazardous triphenylmethane dye, Brilliant Blue FCF. Both the materials were characterized by chemical analysis, IR, DTA, SEM and XRD studies. Their physical characteristics like surface area, porosity, density and loss on ignition were also determined. The adsorption of the dye over both materials was achieved under different pH, adsorbate concentration, sieve size, adsorbent dosage, contact time and temperature, etc. conditions. For both the systems Langmuir and Freundlich adsorption isotherm models were applied and, based on these models, useful thermodynamic parameters were calculated. For both the adsorbents, the kinetic measurements indicate that the adsorption process follows first order kinetics and film diffusion and particle diffusion mechanisms are operative at lower and higher concentrations, respectively, in each case. By percolating the dye solution through fixed-bed columns the bulk removal of the Brilliant Blue FCF was carried out and necessary parameters were determined to find out the percentage saturation of both the columns. Recovery of Brilliant Blue FCF was made by eluting dilute NaOH of pH 11 through each column. PMID- 16040037 TI - Gold nanoparticles prepared using polyethylenimine adsorbed onto montmorillonite. AB - Polyethylenimine-modified montmorillonite (N-MMT) was used to prepare gold nanoparticles, where the montmorillonite (MMT) acted as a solid support to retain the conformation of polyethylenimine (PEI), and the amino groups of PEI were used simultaneously to both complex and reduce the gold ions. From the results of X ray diffraction, it is apparent that the reduction of gold ions occurs primarily on the MMT surface. In the presence of MMT, the formation of a flattened configuration on the clay instead of stretched-out ethylenimine segments of PEI results in the formation of smaller gold particles. With a higher acidification ratio, the recharging of the MMT surface with positive ammonium ionic sites of PEI is likely to prevent the flocculation of clay and thus facilitate the reduction of gold. The rate of gold reduction with N-MMT is faster at low pH values, this being in contrast to the usual trend observed for the reduction of gold ions. The use of PEI adsorbed onto MMT has been shown to be able to act simultaneously as both a protective template and as a reducing agent, thereby greatly simplifying the process for preparing gold nanoparticles. PMID- 16040038 TI - Wormlike micelles and microemulsions in aqueous mixtures of sucrose esters and nonionic cosurfactants. AB - A study of the phase and rheological behavior of sucrose hexadecanoate (C16SE)/cosurfactant/water systems in the presence of solubilized oil, using complementary techniques such as dynamic light scattering and small angle X-ray scattering, is presented. Viscoelastic wormlike micellar solutions are found when a nonionic lipophilic cosurfactant is added to C16SE aqueous systems. Contrary to previous reports, the effect of oil solubilization on these wormlike micelles is not unique and depends on several factors. Linear alkyl chain oils that tend to solubilize in the micellar core have a disrupting effect, decreasing the relaxation time and the viscosity of the systems. This effect is larger as the molecular volume of oil increases and as the solubility of the cosurfactant in oil increases. On the other hand, oils that penetrate in the palisade layer, such as p-xylene, induce micellar growth and have a thickening effect at a given micellar composition. Thermodynamic considerations are used to explain the experimental results. PMID- 16040039 TI - Pioneer studies on HCl and silylation treatments of chrysotile. AB - In the light of some recent studies on organosilicon derivatives of HCl-leached chrysotile, pioneer works not cited in this area are chronologically and briefly commented on, emphasizing the infrared spectroscopy (FTIR-PAS) characterization of such chrysotile products. The latter have opened a far-reaching potential for the development of, e.g., highly stable hydrophobic or organophilic materials, including fibrous sheet polymers. PMID- 16040040 TI - Kinetic and isotherm studies of Cu(II) adsorption onto H3PO4-activated rubber wood sawdust. AB - Adsorption of Cu(II) from aqueous solution onto H(3)PO(4)-activated carbon using rubber wood sawdust (RSAC) was investigated in a batch system. Kinetic and isotherm studies were carried out by considering the effects of various parameters, such as initial concentration, contact time, pH, and temperature. The optimal pH value for Cu(II) adsorption onto RSAC was found to be 6.0. Thermodynamic parameters such as standard Gibbs free energy (DeltaG(0)), standard enthalpy (DeltaH(0)), and standard entropy (DeltaS(0)) were evaluated by applying the Van't Hoff equation. The thermodynamics of Cu(II) adsorption onto RSAC indicates its spontaneous and exothermic nature. Langmuir, Freundlich, and Temkin isotherms were used to analyze the equilibrium data at different temperatures. The Langmuir isotherm fits the experimental data significantly better than the other isotherms. Adsorption kinetics data were tested using pseudo-first-order, pseudo-second-order, and intraparticle diffusion models. Kinetic studies showed that the adsorption followed a pseudo-second-order reaction. The initial sorption rate, pseudo-first-order, pseudo-second-order, and intraparticle diffusion rate constants for different initial concentrations were evaluated and discussed. Adsorption mechanism studies revealed that the process was complex and followed both surface adsorption and particle diffusion. The rate-controlling parameter and effective diffusion coefficient were determined using the Reichenberg plot. It was found that the adsorption occurs through film diffusion at low concentrations and at higher concentration the particle diffusion becomes the rate-determining step. PMID- 16040041 TI - Gold-conductive polymer nanoparticles: a hybrid material with enhanced photonic reactivity to environmental stimuli. AB - We have designed a simple synthetic procedure to encapsulate colloidal gold nanoparticles by electrostatic adsorption with water-soluble poly(aniline-2 carboxylic acid). The composite nanoparticles are stable in aqueous buffer and retain the respective optical reactivity of the gold colloid to refractive index increases, and of the conductive polymer to pH changes and oxidoreduction. The new composite displays, however, significant enhancements in photonic performance when compared to the individual components, which seem to result from electronic interplay between the two materials in the hybrid structure. The enhanced photonic reactivity of the composite structure offers new opportunities for biosensing application. PMID- 16040042 TI - Fragmentation of wastewater sludge floc by planar ice front. AB - Floc size has substantial impact on sludge dewaterability, which might be increased or reduced after freezing and thawing. It is commonly assumed that floc size would be increased by low-speed freezing, with a planar ice front rejecting most flocs ahead of it to form large aggregates. We demonstrate in this work that an advancing planar ice front can not only engulf an activated sludge floc of size 3030 mum, but also fragment it. During floc freezing, when the ice engulfed a thin layer of floc, the latter would be pulled apart vertically by the action of the former. This particular portion of floc was then axially elongated and fixed in the frozen layer, with accumulated force pushing upward. In the present test the floc's vertical length was increased by over 92% and its width decreased by 37% over freezing. The force measurement and floc morphology tracking revealed that the force gradient that pulled apart the floc was 0.0027 N/m. The floc under investigation was fragmented at the point where the normal stress acting on the interior network exceeded 8 Pa. PMID- 16040043 TI - Effects of self-assembly process of latex spheres on the final topology of macroporous silica. AB - This paper surveys the topology of macroporous silica prepared using latex templates covering the submicrometric range (0.1-0.7 mum). The behavior of latex spheres in aqueous dispersion has been analyzed by dynamic light scattering (DLS) measurement indicating the most appropriate conditions to form well-defined cubic arrays. The optical behavior of latex spheres has been analyzed by transmittance and reflectance measurements in order to determine their diameter and filling factor when they were assembled in bidimensional arrays. Macroscopic templates have been obtained by a centrifugation process and their crystalline ordering has been confirmed by porosimetry and scanning electron microscopy. These self assembled structures have been used to produce macroporous silica, whose final topology depends on the pore size distribution of the original template. It has been seen that latex spheres are ordered in a predominant fcc arrangement with slipping of tetragonal pores due to the action of attractive electrostatic interactions. The main effect is to change the spherical shape of voids in macroporous silica into a hexagonal configuration with possible applications to fabricate photonic devices with novel optical properties. PMID- 16040044 TI - Multistep adsorption of anionic dyes on silica/chitosan hybrid. 1. Comparative kinetic data from liquid- and solid-phase models. AB - In this work, a hybrid silica/chitosan was synthesized and characterized by nitrogen elemental analysis and thermal analysis (TG, DTG, DTA, and DSC) and BET surface area. The hybrid was used in adsorption studies of two anionic dyes from aqueous solutions. A rise of temperature accelerates mass transfer of dyes into the hybrid. However, the maximum adsorption capacities reach similar values from 25 to 55 degrees C. The kinetic data were first evaluated in relation to the decrease of the time-related residual concentration of the dyes in solution, where the second-order model has presented the best fitting. The solid-phase interaction of dye data presents a rough fitting to the traditional first-order Lagergren kinetic model. However, a modified Avrami kinetic equation was successfully fitted to the kinetic quantities, where from five to seven kinetic regions were found. A pore-diffusion model has also demonstrated that the diffusion is the rate-controlling interaction mechanism. However, the experimental-calculated comparative values are the best way to evaluate a specific aqueous- or solid-phase kinetic model. PMID- 16040045 TI - The effects of prior knowledge on children's memory and suggestibility. AB - In this study, 5- and 6-year-olds were read a story and asked to recall its details. Two independent factors-prestory knowledge and poststory suggestions were crossed to examine the effects on children's story recall. The results indicated that prestory social knowledge about the story protagonist as well as academic knowledge relating to the content of the story influenced the accuracy of children's recall immediately after the story presentation. Following the suggestive interview, children reported interviewer-provided social and academic misinformation to a greater extent when the misinformation was consistent with their prior knowledge. In contrast, children were more likely to refute misinformation that contradicted their academic knowledge. These findings are discussed in terms of the mechanisms underlying the knowledge-memory and knowledge-suggestibility linkages. PMID- 16040046 TI - Syllable splitting in literate and preliterate Hebrew speakers: onsets and rimes or bodies and codas? AB - This study examined consonant-vowel-consonant (CVC) syllable splitting among literate (Grade 2) and preliterate (kindergarten) Hebrew speakers. Consideration of both the architecture of Hebrew orthography and phonology led to the prediction that a body-coda rather than an onset-rime subdivision would predominate. Structured and unstructured tasks confirmed the claim that there exists a subsyllabic, supraphonemic level of phonological awareness that is more accessible than individual phonemes. However, as predicted, the syllable body rather than the rime was found to be the more accessible biphonemic unit. Moreover, this preference did not appear to be solely the product of orthographic structure; rather it was also inherent in spoken phonology. Access to single phonemes, in contrast, shifted from an early preliteracy advantage for (monophonemic) onsets to a literacy-based preference for codas. PMID- 16040047 TI - Preparation of an immunoadsorbent coupled with a recombinant antigen to remove anti-acetylcholine receptor antibodies in abnormal serum. AB - An immunoadsorbent that removes anti-acetylcholine receptor antibodies (AChRAb) in abnormal serum of myasthenia gravis (MG) patient was efficiently prepared by an expression product, the functional fragment of AChR(alpha205) fused with maltose binding protein (MBP). The ligand can then covalently bind to amylose resin through MBP fusion protein. It was shown from the result of this study with anti-AChR mice sera that the removal rate of AChRAb on this immunoadsorbent reached 87+/-10% (mean value of 10 mice) and the maximally binding capacity of AChRAb was approximately 260 microg/g immunoadsorbent (wet weight). Moreover, the immunoadsorption test of sera in two MG patients indicated that about 90% and 96% of abnormal AChRAb could be eliminated, while other serum components such as albumin, IgG, IgM and IgA only dropped 18%, 35%, 22%, 15% and 24%, 27%, 15%, 12%, respectively, for two MG patient sera. It is anticipated from this study that the immunoadsorbent reported here could, with further development, find its clinical application for removal of AChRAb from patient serum. PMID- 16040048 TI - PCR-RFLP genotyping for exon 1 and promoter region mutations of the human mannose binding lectin (MBL-2) gene. AB - The Mannose Binding Lectin (MBL) plays an important role in innate immunity and its genetic deficiencies are associated with frequent and prolonged infections. Serum MBL determination may not accurately detect acute phase protein levels and it is also difficult to detect dysfunctional protein. Genotyping of the exon 1 and promoter regions in the MBL gene will provide useful information on the presence of deficiencies in patients. A reproducible PCR-RFLP method is described to accurately detect genotypes of exon 1 and polymorphic haplotypes of the promoter region in the MBL gene. PMID- 16040049 TI - Characterization of three ascovirus isolates from cotton insects. AB - Three new ascovirus isolates were discovered from lepidopteran larvae in cotton fields in Blackville, South Carolina, USA, and were named TnAV-2c, TnAV-2d, and HvAV-3f. TnAV-2c and TnAV-2d were compared by restriction endonuclease (REN) profiles and found to be similar. HvAV-3f was isolated from Helicoverpa zea, and bears remarkable dissimilarity in REN profiles to the reported SeAV-5a from Spodoptera exigua but DNA hybridization shows they are closely related. Major capsid protein (MCP) and delta DNA polymerase from the three isolates were sequenced, which suggests the three isolates are novel. Phylogenetic analyses showed that TnAV-2c is distantly related to other lepidopteran ascoviruses. HvAV 3f and SeAV-5a may also be variants of the same species based on Southern, Western, and MCP/DNA polymerase gene sequence analyses. High levels of TnAV-2 infection in an H. zea population (as high as 74%) were recorded in a cotton field in Blackville, SC. Observations in this field showed that infection by ascovirus altered the feeding behavior of H. zea larvae. PMID- 16040050 TI - Development of novel method of non-viral efficient gene transfer into neonatal cardiac myocytes. AB - To establish new treatment for cardiovascular disease, the development of safe and highly efficient vectors is necessary. Especially, non-viral vectors are considered to be ideal for human gene therapy, since recent adverse events with retroviral or adenoviral vectors have highlighted the issue of safety. Although we previously reported safety and high efficiency of HVJ-liposome method, we have modified the envelope of HVJ (Sendai virus). In this novel non-viral vector, the envelope of HVJ alone was utilized as a carrier to deliver proteins, genes and oligodeoxynucleotides (ODN). Thus, we optimized the transfection efficiency of HVJ-envelope vector into neonatal cardiac myocytes in this study, since cardiac myocytes is one of the most difficult cells to be transfected. HVJ-envelope, obtained after complete destruction of HVJ genome, containing FITC-labeled ODN or luciferase plasmid was incubated with cardiac myocytes. In addition, the concentration of protamine sulfate was modified (0-700 microg/ml) to increase transfection efficacy. Without HVJ-envelope vector, few cells showed fluorescence, whereas most cells demonstrated fluorescence with HVJ-envelope vector. Consistent with the high transfection efficiency of ODN, high luciferase activity was also detected using HVJ-envelope vector. Moreover, the transfection efficiency varied according to the concentration of protamine sulfate. No obvious cytotoxicity was observed in cells transfected with HVJ-envelope vector. The present study demonstrated the development of a highly efficient novel non-viral vector for cardiac myocytes, suggesting that further development may provide a new useful tool for research and clinical gene therapy in the field of cardiovascular disease. PMID- 16040051 TI - Cyclooxygenase-2 promotes early atherosclerotic lesion formation in ApoE deficient and C57BL/6 mice. AB - Cyclooxygenase (COX) 2 is expressed in atherosclerotic lesions. We have previously reported that selective inhibition of COX-2 reduces early atherosclerosis in LDLR deficient mice. To examine the role of COX-2 in atherosclerosis in other mouse models, we studied the effects of selective COX-2 inhibition (by rofecoxib and NS-398) and nonselective COX inhibition (by indomethacin) on early atherosclerotic lesion formation in apolipoprotein E deficient (apoE(-/-)) mice. Selective COX-2 and nonselective COX inhibition reduced atherosclerosis in female apoE(-/-) mice by 35-38% and 38-51% in the proximal and en face aortas, respectively. Next we investigated the role of macrophage COX-2 by transplanting COX-2(-/-) fetal liver cells into C57BL/6 mice and challenging the mice with an atherogenic diet. Genetic deletion of COX-2 from hematopoietic cells reduced atherosclerosis by 51%. In addition, LPS activated COX-2(-/-) macrophages had decreased expression of monocyte chemoattractant protein-1 (MCP-1) and tumor necrosis factor-alpha (TNFalpha). The results demonstrate that selective inhibition of COX-2 and elimination of COX-2 from macrophages significantly reduces early atherosclerotic lesion formation in apoE deficient and C57BL/6 mice. These results are compatible with COX-2 expression by macrophages having a proatherogenic role, and support the potential of anti inflammatory therapeutic approaches for atherosclerosis. PMID- 16040052 TI - Predicting membrane protein type by functional domain composition and pseudo amino acid composition. AB - Given the sequence of a protein, how can we predict whether it is a membrane protein or non-membrane protein? If it is, what membrane protein type it belongs to? Since these questions are closely relevant to the function of an uncharacterized protein, their importance is self-evident. Particularly, with the explosion of protein sequences entering into databanks and the relatively much slower progress in using biochemical experiments to determine their functions, it is highly desired to develop an automated method that can be used to give a fast answers to these questions. By hybridizing the functional domain (FunD) and pseudo-amino acid composition (PseAA), a new strategy called FunD-PseAA predictor was introduced. To test the power of the predictor, a highly non-homologous data set was constructed where none of proteins has 25% sequence identity to any other. The overall success rates obtained with the FunD-PseAA predictor on such a data set by the jackknife cross-validation test was 85% for the case in identifying membrane protein and non-membrane protein, and 91% in identifying the membrane protein type among the following 5 categories: (1) type-1 membrane protein, (2) type-2 membrane protein, (3) multipass transmembrane protein, (4) lipid chain-anchored membrane protein, and (5) GPI-anchored membrane protein. These rates are much higher than those obtained by the other methods on the same stringent data set, indicating that the FunD-PseAA predictor may become a useful high throughput tool in bioinformatics and proteomics. PMID- 16040053 TI - Predicting copper toxicity in estuarine and marine waters using the Biotic Ligand Model. AB - The Biotic Ligand Model (BLM) has proven efficient in predicting the toxicity of a variety of metals to freshwater organisms. Consequently, the US EPA has proposed its use for calculating freshwater copper criteria. This study evaluates the BLM for use in estuarine and marine waters. Studies were conducted using the bivalve, Mytilus sp. and 48-h embryo-larval development chronic estimator test methods. These are the most sensitive taxa and test in the US EPA saltwater copper criteria database. Samples from five locations around the USA were tested. There is a strong relationship between measured and BLM predicted copper EC50s (log transformed data, r(2)=0.76, p<0.001, n=44). The BLM predicted within a factor +/-2 of measured EC50s in 41 of 44 cases. However, the BLM tends to predict lower EC50s when measured EC50s are approximately < or =10 microg Cu/L. This may be due to limitations of the metal-dissolved organic matter interaction model. PMID- 16040054 TI - Levels and temporal trends of PCDD/PCDFs and non-ortho PCBs in ringed seals from East Greenland. AB - The levels and temporal trend of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) and non-ortho substituted PCBs (c-PCBs, i.e. CB77, CB126 and CB169) were determined in ringed seal blubber from central East Greenland collected in 1986, 1994, 1999 and 2003, respectively. Since 1986 the concentrations of PCDDs, PCDFs and c-PCBs all show a decreasing trend. The annual decreases were estimated to 5.2% and 5.3% for pg/g WHO-TEQ ww of PCDD/Fs and c PCB, respectively. The annual median concentrations of PCDDs ranged from 5.4 to 24.4 pg/g WHO-TEQ ww and those of PCDFs from 2.5 to 5.1 pg/g WHO-TEQ ww. Compared to PCDD/Fs concentrations in ringed seals from other Arctic areas the levels of PCDD/Fs found in 1986 were the highest recorded. The annual median concentrations of c-PCBs decreased 24.2 to 9.1 pg/g WHO-TEQ ww. The levels of c-PCBs observed in 1986 are similar to levels found in ringed seals from Svalbard in 1990 and from eastern Hudson Bay in 1989-1992. The dominant and most TEQ-contributing PCDD congener was 1,2,3,7,8-PeCDD. CB126 was the dominating and most TEQ-contributing c-PCB congener. The concentrations of PCDDs, PCDFs and c-PCBs were highly significantly inter-correlated. Principal component analysis of the PCDD/PCDF congeners and c-PCBs was performed to analyse the pattern of compounds during time. PMID- 16040055 TI - Susceptibility to oxidative stress of mussels (Mytilus galloprovincialis) in the Venice Lagoon (Italy). AB - The aim of this study was to evaluate the susceptibility to pollutant mediated oxidative stress of the Mediterranean mussel Mytilus galloprovincialis in the Venice lagoon (Italy). In June 2003, mussels from a farm were transplanted to eight sites in the lagoon for five weeks. Oxidative stress responses were measured by: (i) total oxyradical scavenging capacity (TOSC) assay, for an overall evaluation of the oxidative stress response capability; (ii) catalase (CAT), as a key enzyme involved in the antioxidant defence system; (iii) malondialdehyde (MDA), as an indicator of lipid peroxidation, to evaluate an oxidative damage; (iv) metallothioneins (MTs), as they play a role in the antioxidant defence. The TOSC analysis revealed a reduced capability to eliminate: (i) peroxyl radical in mussels transplanted at Palude della Rosa, Valle Millecampi and Chioggia; (ii) hydroxyl radical at Campalto and Valle Millecampi; (iii) peroxynitrite at Valle Millecampi. Inhibition in CAT activity, observed in all the monitored sites, confirms the presence of an oxidative pressure in transplanted mussels. In addition, Pearson correlation analysis was performed in order to observe possible links between the various parameters. The PCA was a powerful tool to discriminate impacted sites, suggesting that the mussels transplanted throughout the Venice lagoon were subjected to different levels of oxidative pressure. Furthermore, it provided an easy and useful tool to summarize the obtained results. PMID- 16040056 TI - Cost/benefit analysis of a benthic monitoring programme of organic benthic enrichment using different sampling and analysis methods. AB - To investigate the combined effects of decreasing taxonomic resolution (i.e. species, family, phylum), the use of different mesh-size (1.0 mm and 0.5 mm) and the type of samplers used (van Veen vs. corers taken by divers) on the quality of data obtained, a comparative study was undertaken with the overall aim of identifying cost efficient methods for routinely monitoring the ecological change caused by Mediterranean fish farming. The results clearly showed that information loss was relatively low as data were aggregated at higher taxonomic levels, particularly up to the level of family or even order. It was also found that the extra information gained by sieving samples through a 0.5 mm sieve did not improve the ability to distinguish the potentially impacted sites from the control stations. Finally, it was found that a relatively large proportion of the available information concerning the community structure such as abundance, biomass or diversity is lost when sampling is carried out with corers. A cost/benefit ratio analysis for the two sampling and the two sieving methods showed minimal values for the van Veen samples (for both sieve fractions) at the family level, indicating that analysis at this level gives the best balance between precision of the results and decrease in taxonomic effort. However, if the time needed to sort the samples is included in the analysis, then samples taken with corers using a 0.5 mm sieve and identified to families seems like a good compromise between precision and cost. PMID- 16040057 TI - Okadaic acid, a causative toxin of diarrhetic shellfish poisoning, in green lipped mussels Perna viridis from Hong Kong fish culture zones: method development and monitoring. AB - Green-lipped mussels (Perna viridis) were collected from seven fish culture zones (FCZs) in Hong Kong and analyzed for okadaic acid (OA). A conventional HPLC method was modified by incorporating a proteinase K digestion step. Results suggest that a higher recovery (2.5 times higher) of OA was obtained from spiked samples after the addition of 1.08 mg proteinase K in comparison with samples incubated without the proteolytic enzyme. For the hepatopancreas (HP) of individual field-collected mussels, the additional digestion step can enhance OA extraction by 3.1 times. Spatial and temporal variations in OA concentrations in the mussels from various FCZs were investigated. The highest concentration of OA in mussel HP samples was 1164.9 ng/g HP wet wt. With respect to OA concentrations in whole mussel tissues from seven sites and four seasonal samplings, the concentrations were between 70.0 and 131.0 ng/g wet wt., which did not exceed the generally recognized international regulatory criteria (>200 ng/g) for OA. PMID- 16040058 TI - Butyltin compounds in a sediment core from the old Tilbury basin, London, UK. AB - Sections from a sediment core taken from the River Thames were analysed for butyltin species using gas chromatography with species-specific isotope dilution mass spectrometry. Results demonstrated that in most samples tributyltin concentrations of 20-60 ng/g accounted for <10% of the total butyltin species present, which is in agreement with data from other sediment samples which were historically contaminated with tributyltin. Vertical distribution of the organotin residues with depth throughout the core, with data on organochlorine compounds and heavy metals allowed for the construction of a consistent hypothesis on historical deposition of contaminated sediments. From this it was possible to infer that the concentrations of tributyltin in sediments deposited during the early 1960s were in the order of 400-600 microg/g by using degradation rate constants derived by other workers. Such values fall well within the range quoted for harbour sediments in the literature. PMID- 16040059 TI - Atrazine effects on meiobenthic assemblages of a modular estuarine mesocosm. AB - Atrazine is a widely used herbicide in the US found at levels ranging from <10 ng/L to 62.5 microg/L in estuaries throughout the southeast. Effects of atrazine on estuarine meiobenthic assemblages chronically exposed to environmentally relevant concentrations are unknown. The purpose of our research was to assess effects of atrazine on meiobenthos at concentrations near the proposed USEPA SWQC (26 microg/L) using modular estuarine salt marsh mesocosms as a field surrogate. Indigenous copepod and nematode densities were assessed after 28 days of exposure in transplanted colonization chambers. Cluster analysis showed a group characterized by low copepod densities, mostly atrazine exposed chambers, and a group containing all but one control chamber. The later group included chambers with high densities of the copepods Paronychocamptus wilsoni and Enhydrosoma baruchi. Compared to controls, copepod densities was approximately 70% lower in atrazine chambers, with three of the most common copepod species (E. baruchi, Onychocamptus sp. and P. wilsoni) showing an average of 50-70% reduction in population densities (p<0.05). Although nematode density did not differ between atrazine and control chambers, the nematode-to-copepod ratio was significantly higher in atrazine (9.95+/-7.61; p=0.011) than in control chambers (0.61+/-0.35). Our findings suggest that chronic exposures over multiple generations to atrazine at concentrations near the proposed USEPA SWQC could have significant effects on the abundance and composition of estuarine meiobenthic copepod assemblages. PMID- 16040060 TI - Concentrations of alkylphenols in sediments from the Pearl River estuary and South China Sea, South China. PMID- 16040061 TI - Accumulation of mercury, cadmium, lead and arsenic in swordfish and bluefin tuna from the Mediterranean Sea: a comparative study. PMID- 16040062 TI - The bystander effect-induced formation of micronucleated cells is inhibited by antioxidants, but the parallel induction of apoptosis and loss of viability are not affected. AB - X-rays induce various DNA damages including strand breaks that lead to formation of micronuclei and chromosomal aberrations as well as increased number of apoptotic cells. Similar effects appear when non-irradiated cells are treated with medium collected from cultures of irradiated cells (irradiation conditioned medium - ICM). This phenomenon was termed "bystander effect". A number of studies suggest that bystander effect appears to be associated with up-regulation of oxidative metabolism. We thus compared the effects of antioxidant Vitamins C and E on the frequency of micronuclei and apoptotic cells in both directly irradiated cell cultures and in cultures exposed to ICM. Addition of Vitamins C or E (1-40 microg/ml) to culture medium after exposure to radiation or ICM reduced the frequency of micronuclei in a concentration-dependent manner. These vitamins had no effect on cell viability, clonogenic survival or the frequency of apoptotic cells under both conditions tested. These results show that the bystander effect causes micronucleation in addition to other known effects and suggest that the factors causing micronucleation by X-irradiation, oxidative DNA damage and incomplete repair, are regulated by apoptosis-independent pathways. PMID- 16040063 TI - The antidepressant-like effect of Hypericum caprifoliatum Cham & Schlecht (Guttiferae) on forced swimming test results from an inhibition of neuronal monoamine uptake. AB - A crude (ECH) and a purified cyclohexane extract (HCP) of Hypericum caprifoliatum and their main phloroglucinol derivative (HC1) were evaluated regarding their action on monoaminergic systems, more precisely on dopamine. In rats and mice forced swimming test, ECH and HCP dose-dependently reduced the immobility time. The effect of the highest dose was prevented by a prior administration of either sulpiride or SCH 23390 (D(2) and D(1) dopamine receptor antagonist, respectively). HCP (360 mg/kg) decreased the locomotor activity of mice. ECH (90 mg/kg) caused hypothermia and potentiated apomorphine-induced (16 mg/kg) hypothermia in mice. HCP and HC1 inhibited, in a concentration-dependent and monophasic manner, the [(3)H]-DA, [(3)H]-NA and [(3)H]-5HT synaptosomal uptakes, but did not prevent the binding of specific ligands to the monoamine transporters. Moreover, when tested at the concentrations corresponding to its IC(50) on [(3)H]-DA uptake, HC1 did not induce a significant [(3)H]-DA release, while at a higher concentration (200 ng/ml) it enhanced significantly (by 12%) the synaptosomal DA release. These data suggest that the antidepressant-like effect of H. caprifoliatum on the forced swimming test is due to an increase in monoaminergic transmission, resulting from monoamine uptake inhibition, more potently of dopamine, which may be related to their phloroglucinol contents. PMID- 16040064 TI - Grm5 expression is not required for the oncogenic role of Grm1 in melanocytes. AB - Melanoma is the aberrant proliferation of melanocytes, the cells in the skin responsible for pigment (melanin) production. In its early stages, melanoma can be surgically removed with great success, however, advanced stages of melanoma have a high mortality rate due to the lack of responsiveness to currently available therapies. We have previously characterized a mouse melanoma model, TG 3, which has implicated the ectopic expression of metabotropic glutamate receptor 1 (Grm1, formerly mGluR1), in melanomagenesis and metastasis [Pollock et al., 2003. Melanoma mouse model implicates metabotropic glutamate signaling in melanocytic neoplasia. Nat Genet. 34, 108-112.]. Here we report the characterization of several in vitro cell lines derived from independent mouse melanoma tumors. These cell lines show characteristic phenotypes of transformed melanocytes, and express Grm1, and Grm5 (another metabotropic glutamate receptor), as well as melanocyte-specific protein markers. To investigate the possible role of Grm5 in vivo during melanoma development in our mice, we have crossed Grm5 null mice with TG-3, generating a new line of transgenic mice, TGM. TGMs, which are homozygote knockouts for Grm5 and carry the TG transgene, develop tumors with onset, progression, and metastasis very similar to that described for TG-3. Taken together, these results indicate that Grm1 can act as an oncogene in melanocytes independently of Grm5 expression. PMID- 16040065 TI - Hippocampal dopamine receptors modulate cFos expression in the rat nucleus accumbens evoked by chemical stimulation of the ventral hippocampus. AB - Recently, we have shown that D1 and D2 receptors in the ventral hippocampus (VH) modulate both the locomotor activation and the increase in dopamine (DA) levels in the rat nucleus accumbens (NAc) induced by NMDA stimulation of the VH. In the present study we analyze the possible role of VH D1 and D2 receptors in the modulation of the cFos expression in NAc (core and shell subregions) and in dorsal striatum. This was assessed by immunohistochemical analysis of cFos expression in the rat brains after retro-dialysis application of NMDA (50mM, 10 min) into VH, in absence and in presence of either the D1/D5 receptor antagonist SCH 23390 (100 and 250 microM, 60 min) or the D2 receptor antagonist raclopride (100 and 250 microM, 60 min). NMDA induced a robust increase in the cFos expression in the NAc shell, both in the ipsilateral and contralateral side. No statistically significant increases were observed in the NAc core and in the dorsal striatum. Simultaneous application of SCH 23390 and NMDA into the VH attenuated the NMDA-evoked cFos expression in NAc shell. In contrast, raclopride had no significant effect. Our present results show that the NMDA receptor mediated effects in the VH require D1 receptors and suggest that DA in VH strongly modulates the excitatory outputs from this brain area. PMID- 16040066 TI - Two cyclopeptides from the seeds of Annona cherimola. AB - Bioassay-guided fractionation of cytotoxic of methanol extract of the seeds of Annona cherimola provided two novel cyclic peptides, cherimolacyclopeptide E (1) and cherimolacyclopeptide F (2), which exhibited significant cytotoxic activity against the KB (human nasopharyngeal carcinoma) cell culture system. The peptide 1 and 2 were elucidated by MS/MS fragmentation experiments using a Q-TOF mass spectrometer equipped with an ESI source, extensive 2D NMR analyses and chemical degradation. PMID- 16040067 TI - In vitro effect of recombinant human tumor necrosis factor-alpha on canine neutrophil apoptosis. AB - Neutrophils (polymorphonuclear leukocytes: PMNs) are essential for the host defense against various infections and are often injurious to the host, causing inflammatory diseases where tumor necrosis factor-alpha (TNF-alpha) is suggested to play an important role. Since an effect of TNF-alpha on canine PMN apoptosis has not been studied, canine PMNs were stimulated with recombinant human (rh)TNF alpha in the present study to investigate the effect of TNF-alpha on canine PMN apoptosis. PMN apoptosis and function to produce ROS were assessed by flow cytometry. Delayed apoptosis was observed in the PMNs treated with rhTNF-alpha at 100 ng/ml, accompanied by retention of capability to produce ROS. However, PMN apoptosis was accelerated by rhTNF-alpha combined with cycloheximide. Therefore, it is indicated that TNF-alpha is able to activate anti- and pro-apoptotic pathways in PMNs and that the inhibition of PMN apoptosis by TNF-alpha requires protein synthesis in the PMNs. PMID- 16040068 TI - A high-performance liquid chromatography-tandem mass spectrometry assay of the androgenic neurosteroid 3alpha-androstanediol (5alpha-androstane-3alpha,17beta diol) in plasma. AB - The testosterone metabolite 3alpha-androstanediol (5alpha-androstane-3alpha,17 diol) is a potential GABA(A) receptor-modulating neurosteroid with anticonvulsant properties and hence could act as a key neuromodulator in the central nervous system. However, there is no specific and sensitive assay for quantitative determination of the androgenic neurosteroid 3alpha-androstanediol in biological samples. We have established a liquid chromatography-tandem mass spectrometry (LC MS-MS) assay to measure 3alpha-androstanediol in rat plasma. Standard 3alpha androstanediol added to rat plasma has been successfully analysed with excellent linearity, specificity, sensitivity, and reproducibility. The sensitivity of the method was < 10 ng/ml with a detection limit of 2 ng/ml (6.8 nmol/l) and a linear range of 10-2000 ng/ml. The method was used for the analysis of testosterone induced increase in plasma 3alpha-androstanediol levels in rats. Testosterone produced a dose-dependent elevation in plasma 3alpha-androstanediol, which was almost completely prevented by pretreatment with the 5alpha-reductase inhibitor finasteride, indicating that 3alpha-androstanediol is synthesized from testosterone via a 5alpha-reductase pathway. This LC-MS-MS method allows accurate, high-throughput analysis of 3alpha-androstanediol in small amounts (200 microl) of plasma and possibly other biological samples. PMID- 16040069 TI - The dynamical implications of disease interference: correlations and coexistence. AB - Ecological interference between unrelated diseases, caused by the temporary or permanent removal of individuals susceptible to one disease following infection with another, might be an important mechanism underlying epidemics. In this paper, we explore the potential dynamic consequences of interference by analyzing a two-disease model. By studying the stability domain of the model's equilibria, we find that the stable region of the two-disease endemic state becomes increasingly smaller as the strength of interference (largely determined by the disease-induced mortality) increases. When seasonal changes are included in the transmission rates, the bifurcation structure of the model's periodic cycles reveals that when the two diseases have similar mean transmission rates, multiple attractors in which the two diseases are strongly correlated can coexist, and that when the two diseases have very different mean transmission rates, the one with higher mean transmission rate may determine the dynamics of the system, with the other infection mimicking the behavior. We conclude that ecological interference can have important effects on the dynamical pattern of interacting diseases, the extent of which is determined by the epidemiological features of the diseases, their mean transmission rates in particular. PMID- 16040070 TI - Does quasi-local competition lead to pattern formation in metapopulations? An explicit resource competition model. AB - In metapopulations, competitive interactions may extend beyond the confines of the local population such that members of neighbouring habitat patches affect each other adversely (quasi-local competition). We derive a model for quasi-local competition from first principles, assuming that individuals compete for shared resources and members of a population spend a certain fraction of their foraging time in the adjacent populations. Contrary to the results of Doebeli and Killingback [2003. Theor. Popul. Biol. 64, 397-416], our model does not produce spatial patterns of population densities in homogeneous environments. Quasi-local competition nevertheless contributes to pattern formation by amplifying the effect of heterogeneities in the external environment, and this amplification can be extremely strong when dispersal is absent. We discuss why apparently similar models lead to contrasting results. PMID- 16040071 TI - The impact of mortality on predator population size and stability in systems with stage-structured prey. AB - The relationships between a predator population's mortality rate and its population size and stability are investigated for several simple predator-prey models with stage-structured prey populations. Several alternative models are considered; these differ in their assumptions about the nature of density dependence in the prey's population growth; the nature of stage-transitions; and the stage-selectivity of the predator. Instability occurs at high, rather than low predator mortality rates in most models with highly stage-selective predation; this is the opposite of the effect of mortality on stability in models with homogeneous prey populations. Stage-selective predation also increases the range of parameters that lead to a stable equilibrium. The results suggest that it may be common for a stable predator population to increase in abundance as its own mortality rate increases in stable systems, provided that the predator has a saturating functional response. Sufficiently strong density dependence in the prey generally reverses this outcome, and results in a decrease in predator population size with increasing predator mortality rate. Stability is decreased when the juvenile stage has a fixed duration, but population increases with increasing mortality are still observed in large areas of stable parameter space. This raises two coupled questions which are as yet unanswered; (1) do such increases in population size with higher mortality actually occur in nature; and (2) if not, what prevents them from occurring? Stage-structured prey and stage related predation can also reverse the 'paradox of enrichment', leading to stability rather than instability when prey growth is increased. PMID- 16040072 TI - Analysis of thyroid hormone receptor betaA mRNA expression in Xenopus laevis tadpoles as a means to detect agonism and antagonism of thyroid hormone action. AB - Amphibian metamorphosis represents a unique biological model to study thyroid hormone (TH) action in vivo. In this study, we examined the utility of thyroid hormone receptors alpha (TRalpha) and betaA (TRbetaA) mRNA expression patterns in Xenopus laevis tadpoles as molecular markers indicating modulation of TH action. During spontaneous metamorphosis, only moderate changes were evident for TRalpha gene expression whereas a marked up-regulation of TRbetaA mRNA occurred in hind limbs (prometamorphosis), head (late prometamorphosis), and tail tissue (metamorphic climax). Treatment of premetamorphic tadpoles with 1 nM 3,5,3' triiodothyronine (T3) caused a rapid induction of TRbetaA mRNA in head and tail tissue within 6 to 12 h which was maintained for at least 72 h after initiation of T3 treatment. Developmental stage had a strong influence on the responsiveness of tadpole tissues to induce TRbetaA mRNA during 24 h treatment with thyroxine (0, 1, 5, 10 nM T4) or T3 (0, 1, 5, 10 nM). Premetamorphic tadpoles were highly sensitive in their response to T4 and T3 treatments, whereas sensitivity to TH was decreased in early prometamorphic tadpoles and strongly diminished in late prometamorphic tadpoles. To examine the utility of TRbetaA gene expression analysis for detection of agonistic and antagonistic effects on T3 action, mRNA expression was assessed in premetamorphic tadpoles after 48 h of treatment with the synthetic agonist GC-1 (0, 10, 50, 250 nM), the synthetic antagonist NH-3 (0, 40, 200, 1000 nM), and binary combinations of NH-3 (0, 40, 200, 1000 nM) and T3 (1 nM). All tested concentrations of GC-1 as well as the highest concentration of NH-3 caused an up-regulation of TRbetaA expression. Co-treatment with NH-3 and T3 revealed strong antagonistic effects by NH-3 on T3-induced TRbetaA mRNA up regulation. Results of this study suggest that TRbetaA mRNA expression analysis could serve as a sensitive molecular testing approach to study effects of environmental compounds on the thyroid system in X. laevis tadpoles. PMID- 16040073 TI - Strategies to assess systemic exposure of chemicals in subchronic/chronic diet and drinking water studies. AB - Strategies were developed for the estimation of systemically available daily doses of chemicals, diurnal variations in blood levels, and rough elimination rates in subchronic feeding/drinking water studies, utilizing a minimal number of blood samples. Systemic bioavailability of chemicals was determined by calculating area under the plasma concentration curve over 24 h (AUC-24 h) using complete sets of data (> or =5 data points) and also three, two, and one selected time points. The best predictions of AUC-24 h were made when three time points were used, corresponding to Cmax, a mid-morning sample, and C(min). These values were found to be 103 +/- 10% of the original AUC-24 h, with 13 out of 17 values ranging between 96 and 105% of the original. Calculation of AUC-24 h from two samples (Cmax and Cmin) or one mid-morning sample afforded slightly larger variations in the calculated AUC-24 h (69-136% of the actual). Following drinking water exposure, prediction of AUC-24 h using 3 time points (Cmax, mid-morning, and Cmin) was very close to actual values (80-100%) among mice, while values for rats were only 63% of the original due to less frequent drinking behavior of rats during the light cycle. Collection and analysis of 1-3 blood samples per dose may provide insight into dose-proportional or non-dose-proportional differences in systemic bioavailability, pointing towards saturation of absorption or elimination or some other phenomenon warranting further investigation. In addition, collection of the terminal blood samples from rats, which is usually conducted after 18 h of fasting, will be helpful in rough estimation of blood/plasma half-life of the compound. The amount of chemical(s) and/or metabolite(s) in excreta and their possible use as biomarkers in predicting the daily systemic exposure levels are also discussed. Determining these parameters in the early stages of testing will provide critical information to improve the appropriate design of other longer-term toxicity studies. PMID- 16040074 TI - Synthesis, cellular localization, and quantification of penton-dodecahedron in serotype 3 adenovirus-infected cells. AB - Adenovirus penton is a non-covalent complex composed of the penton base and fiber proteins, localized at the twelve vertices of the icosahedral virus capsid. In cells infected by adenovirus serotype 3 (Ad3), penton is found not only in the virus capsid but also self-assembled in dodecahedra formed through interactions between the twelve penton bases. In this study, the intracellular trafficking of penton proteins from the cytoplasm to the nucleus has been followed, and the nuclear re-arrangement induced by viral infection has been observed by electron microscopy of ultrathin sections. The amount of dodecahedra has been assessed in relation to the number of Ad3 infectious virions produced during the Ad3 replication cycle. It appears that dodecahedra are produced in a large excess over viral infectious particles and that they are located intranuclearly along the nuclear membrane of Ad3-infected cells at late times of infection. PMID- 16040075 TI - The MCV MC159 protein inhibits late, but not early, events of TNF-alpha-induced NF-kappaB activation. AB - Tumor necrosis factor (TNF-alpha) triggers biphasic activation of the NF-kappaB transcriptional regulator. This process consists of an initial, IkappaBalpha mediated transient phase and a later, persistent phase dependent on IkappaBbeta degradation. To presumably interfere with the fulfillment of this immunity associated event in cells infected with the molluscum contagiosum virus (MCV), this pathogen produces the intracellular MC159 protein. To define the mode of action of MC159, the impact of TNF-alpha on HEK 293T cells ectopically expressing the MC159 protein was examined. In this regard, TNF-alpha-induced expression of an NF-kappaB-regulated luciferase reporter gene was partially inhibited by the MC159 protein. This ability was attributed to blockage of the persistent phase of TNF-alpha-induced NF-kappaB activation for the following reasons: (1) the initial phase of NF-kappaB transcriptional activation was not affected by the MC159 protein; (2) the MC159 protein inhibited TNF-alpha-directed degradation of IkappaBbeta, but not IkappaBalpha; and (3) expression of the late NF-kappaB regulated cell genes, TNF-alpha and CCL2, was decreased in the presence of the MC159 protein while transcription of the early NF-kappaB-regulated cell gene, CXCL1, was not altered. Previously reported MC159-RIP interactions appear to be irrelevant for the MC159 inhibitory function. In contrast, MC159-TRAF2 associations are more relevant for inhibitory function since mutant MC159 proteins unable to bind TRAF2 also cannot inhibit TNF-mediated NF-kappaB activation. In vivo, the MC159 protein may act to prolong virus survival by preventing the infected cell from responding to TNF-alpha, ultimately preventing the cellular production of proinflammatory and immunoattractant molecules. PMID- 16040076 TI - Two mink parvoviruses use different cellular receptors for entry into CRFK cells. AB - Mink enteritis virus (MEV) and Aleutian mink disease parvovirus (ADV) are two mink parvoviruses that replicate permissively in Crandell feline kidney (CRFK) cells. We have used this cell model to examine if these two mink parvoviruses use the same cellular receptor. Whereas the cellular receptor for MEV is expected to be the transferrin receptor (TfR), the cellular receptor for ADV has not been clearly identified. We used short hairpin RNAs (shRNAs) produced from plasmids to trigger RNA interference (RNAi), specifically and effectively reducing TfR expression in CRFK cells. TfR expression was reduced to levels undetectable by immunofluorescence in the majority of cells. In viral infection assays, we show that TfR expression was necessary for MEV infection but was not required for ADV infection. Thus, our results demonstrate that TfR is the cellular receptor for MEV, but not the cellular receptor for ADV. The use of two different receptors by MEV and ADV to infect the same cell line is yet another difference between these two parvoviruses that may contribute to their unique pathogenesis in mink. PMID- 16040077 TI - The orientation bandwidth of cyclopean channels. AB - Orientation bandwidths of cyclopean channels were estimated using a notched noise technique. Observers were presented with random dot stereograms depicting a horizontal or vertical target sinusoidal depth modulation and a mask consisting of sinusoidal depth modulations whose orientations flanked that of the target. Masking reduced as the orientation difference between signal and mask increased. The orientation bandwidth of the masking effect was similar to that found for stimuli defined by luminance contrast, and showed no systematic difference for horizontal and vertical targets. These results suggest that the elongated summation found by Tyler, C. W., and Kontsevich, L. L. (2001). Stereoprocessing of cyclopean depth images: Horizontally elongated summation fields. Vision Research, 41, 2235-2243, for horizontal stimuli occurs after a processing non linearity. PMID- 16040078 TI - Accommodation stimulus-response function and retinal image quality. AB - Accommodation stimulus-response function (ASRF) and its relationship to retinal image quality were investigated using a modified wavefront sensor. Ten subjects were presented with six vergence stimuli between 0.17 D and 5 D. For each vergence distance, ocular wavefronts and subjective visual acuity were measured. Wavefronts were analysed for a fixed 3-mm pupil diameter and for natural pupil sizes. Visual Strehl ratio computed in the frequency domain (VSOTF) and retinal images were calculated for each condition tested. Subjective visual acuity was significantly improved at intermediate vergence distances (1D and 2D; p < 0.01), and only decreased significantly at 5 D compared with 0.17 D (p < 0.05). VSOTF magnitude was associated with subjective visual acuity and VSOTF peak location correlated with accommodation error. Apparent accommodation errors due to spherical aberration were highly correlated with accommodation lead and lag for natural pupils (R(2) = 0.80) but not for fixed 3-mm pupils (R(2) < 0.00). The combination of higher-order aberrations and accommodation errors improved retinal image quality compared with accommodation errors or higher order aberrations alone. Pupil size and higher order aberrations play an important role in the ASRF. PMID- 16040079 TI - Learning top-down gain control of feature selectivity in a recurrent network model of a visual cortical area. AB - We propose that the effects of attentional top-down modulations observed in the visual cortex reflect the simple strategy of strengthening currently relevant pathways in a task-dependent manner. To exemplify this idea, we set up a network model of a visual area and simulate the learning of a context-dependent 'go/no go'-task. The model learns top-down gain-modulations of sensory representations based on reinforcements received from the environment. We also discuss how this idea relates to alternative interpretations like optimal coding hypotheses. PMID- 16040080 TI - Spatial interactions reveal inhibitory cortical networks in human amblyopia. AB - Humans with amblyopia have a well-documented loss of sensitivity for first-order, or luminance defined, visual information. Recent studies show that they also display a specific loss of sensitivity for second-order, or contrast defined, visual information; a type of image structure encoded by neurons found predominantly in visual area A18/V2. In the present study, we investigate whether amblyopia disrupts the normal architecture of spatial interactions in V2 by determining the contrast detection threshold of a second-order target in the presence of second-order flanking stimuli. Adjacent flanks facilitated second order detectability in normal observers. However, in marked contrast, they suppressed detection in each eye of the majority of amblyopic observers. Furthermore, strabismic observers with no loss of visual acuity show a similar pattern of detection suppression. We speculate that amblyopia results in predominantly inhibitory cortical interactions between second-order neurons. PMID- 16040081 TI - Studies of selenium in environmental samples and synthetic mixtures by spectrophotometry. AB - A rapid, highly sensitive and selective spectrophotometric method for the determination of traces of selenium(IV) is described. The method is based on oxidation of p-nitroaniline by selenium(IV) followed by coupling reaction with N (1-naphthalene-1-yl)ethane-1,2-diamine dihydrochloride (NEDA) in neutral medium to give red colored derivative with lambda(max) 515 nm and is stable for more than 10 days at 35 degrees C. Beer's law is obeyed for selenium(IV) in the concentration range of 0.02-3.2 microg ml(-1) at the wavelength of maximum absorption. The optimum reaction conditions and other analytical parameters were investigated to enhance the sensitivity of the present method. The detailed study of various interferences made the method more selective. The proposed method was successfully applied to the analysis of selenium in polluted water, natural water samples, plant material, soil samples, and synthetic mixtures. The results obtained were agreed with the reported methods at the 95% confidence level. The performance of proposed method was evaluated in terms of Student's t-test and Variance ratio f-test which indicates the significance of proposed method over reported method. PMID- 16040082 TI - Sensitive and quantitative, 10-min immunofluorometric assay for D-Dimer in whole blood. AB - INTRODUCTION: Normal concentrations of D-Dimer can be used to exclude venous thromboembolism (VTE). However, methods for sensitive and quantitative D-Dimer measurements at the point-of-care (POC) are still limited. MATERIALS AND METHODS: We developed a 10-min, non-competitive immunofluorometric assay for D-Dimer in citrated whole blood and plasma using pre-dispensed reagents dried in single assay wells. The simple, automated assay procedure comprises a 1:50 sample dilution, one-step incubation, washing, and time-resolved fluorometric measurement directly from the wet well surface. RESULTS: The limits of detection (background + 3SD) and quantification (CV <15%) were 0.05 and 0.2 mg/L D-Dimer, respectively, and the assay was linear up to 400 mg/L. Correlations to Roche TinaQuant (r=0.726, n=200) and Biopool Auto.Dimer (r=0.190, n=149) were carried out using citrated plasma. Diagnostic sensitivity, specificity, and negative (NPV) and positive (PPV) predictive values were 98.7%, 64.4%, 99.1% and 55.1%, and 92.2%, 81.0%, 95.9% and 68.3%, respectively, using cut-off values of 0.6 and 1.0 mg/L, respectively, in outpatients with deep vein thrombosis (DVT) and/or pulmonary embolism (PE) (n=77) compared with outpatients with various other diseases (n=174). The within- and between-run CVs near the cut-off values were < or =10% in both whole blood and plasma. The 95th percentile upper range in apparently healthy individuals was 0.68 mg/L of whole blood (n=101). CONCLUSIONS: The high sensitivity and NPV suggest that the rapid immunofluorometric assay could be valuable for rapid exclusion of VTE in outpatients. With appropriate cut offs, the assay could potentially be used as a stand-alone test or combined with clinical probability assessment, but further studies are required. PMID- 16040083 TI - From funny current to current physiome. PMID- 16040084 TI - Is patient age a factor in the occurrence of prostate-specific antigen bounce phenomenon after external beam radiotherapy for prostate cancer? AB - OBJECTIVES: To evaluate the effect of patient age on the occurrence of prostate specific antigen (PSA) "bounce" after external beam radiotherapy (EBRT) for prostate cancer. METHODS: In this study, 964 patients received EBRT alone for prostate cancer between April 1987 and January 1998 who had been followed for at least 12 months. Prostate-specific antigen values were obtained every 3 to 6 months after radiotherapy. Median overall follow-up was 48 months. Prostate specific antigen bounce was defined as an initial increase in serum PSA of at least 0.5 ng/mL, followed by a decrease to prebounce baseline serum PSA values, all within 60 months after EBRT. Biochemical failure was defined as three consecutive increases in posttreatment PSA concentration after achieving a nadir. Multivariate Cox regression analysis was performed to evaluate the influences of age, pretreatment PSA concentration, Gleason score (determined at biopsy), clinical T stage classification, and radiation dose on PSA bounce-free survival and biochemical disease-free survival, with P < 0.05 considered statistically significant. RESULTS: Twelve percent of the patients developed a PSA bounce. Age was not associated with the occurrence of a PSA bounce (P = 0.63), the magnitude of the PSA bounce (P = 0.90), or the duration of the PSA bounce (P = 0.39). Patients who had PSA bounce had a statistically significant higher biochemical disease-free survival than those who did not (P = 0.00004). CONCLUSIONS: In our study, age was not predictive of PSA bounce. However, younger patients with a rising PSA after radiotherapy should be followed closely for evidence of biochemical failure. PMID- 16040085 TI - Utility of saturation biopsy to predict insignificant cancer at radical prostatectomy. AB - OBJECTIVES: To determine whether potential candidates for watchful waiting have undersampling of more substantial cancer. METHODS: A total of 103 men were studied, who were predicted to have insignificant cancer in their radical prostatectomy (RP) specimen. All had limited cancer on routine needle biopsy (no core with more than 50% involvement; Gleason score less than 7, and fewer than 3 cores involved) with a serum prostate-specific antigen density of 0.15 or less. Insignificant tumor at RP was considered organ-confined tumor, no Gleason pattern 4 or 5, and a tumor volume of less than 0.5 cm3. Saturation biopsy (average 44 cores) and an alternate biopsy saturation scheme with one half the number of cores using an 18-gauge Biopty gun was performed in the pathology laboratory on totally embedded and serially sectioned RP specimens. RESULTS: Of the tumors, 97% were organ confined. The RP Gleason score was less than 7 in 84% of the cases. The RP tumor volume was 0.01 to 2.39 cm3 (median 0.14). Of the cancer specimens, 71% were insignificant and 29% had been incorrectly classified before surgery using standard biopsy schemes. Using the full saturation biopsy scheme, if we predicted significant cancer, the probability of having insignificant cancer was only 11.5% (false-positive rate). If the model predicted insignificant cancer, the probability of significant cancer was also only 11.5% (false-negative rate; sensitivity 71.9% and specificity 95.8%). Using the alternate biopsy sampling scheme, the false-positive rate was 8% and the false-negative rate was 11.4% (sensitivity 71.9% and specificity 97.1%). CONCLUSIONS: Saturation biopsy provides accurate predictability of prostate tumor volume and grade to select suitable candidates for watchful waiting therapy. PMID- 16040086 TI - Long-term results of suprapubic bladder neck closure for treatment of the devastated outlet. AB - OBJECTIVES: To assess the long-term success of suprapubic bladder neck closure in patients with irreparably damaged bladder outlets. METHODS: A cohort of 35 patients with intractable urinary incontinence secondary to severe posterior urethral/bladder neck damage underwent suprapubic bladder neck closure. Patients were assessed with regard to the success of procedure, as well as early and late complications. RESULTS: With a mean follow-up of 79 months (range 12 to 164), suprapubic bladder neck closure was successful in 29 (83%) of 35 patients. One revision of the bladder neck improved the success rate to 94% (33 of 35). Early and late complications, excluding bladder neck fistula, were reported in 3 (9%) and 5 (14%) of 35 patients, respectively. CONCLUSIONS: High success and acceptable complication rates can be achieved with suprapubic bladder neck closure for the treatment of severe urinary incontinence secondary to a devastated bladder outlet. PMID- 16040087 TI - Survey from skills-based hands on learning courses demonstrates increased laparoscopic caseload and clinical laparoscopic suturing. AB - OBJECTIVES: To evaluate the impact of the American Urological Association Hands on Laparoscopy course on the participants' practices. Many urologists without fellowship training perform laparoscopy, but do not advance beyond hand-assisted, extirpative laparoscopy. The American Urological Association Hands on Laparoscopy course was designed to help these practitioners advance their skills. METHODS: A total of 68 urologists, aged 31 to 61 years (mean 46.6), participated in one of the four courses given between August 2002 and March 2004. The 2-day course included performing standardized tasks under videotape analysis and participating in porcine and pelvic trainer laboratory sessions. Surveys were sent by regular and electronic mail in September 2004 to assess the courses' impact. The mean follow-up was 15.2 months (range 7 to 25). RESULTS: Of the 68 surveys mailed, 54 were returned (79%). Most respondents were in private practice and had had prior experience with extirpative laparoscopy. Of the respondents, 41 (76%) reported that their laparoscopic practice had expanded after taking the course, with 34% performing at least 2 cases per month. Also, 33 respondents (61%) reported performing laparoscopic suturing after taking the course, with 35% having sutured a bleeding vessel. Of the respondents, 85% reported that the video mentoring during the course was helpful. Of those who purchased a pelvic trainer, 90% reported practicing on it regularly. CONCLUSIONS: The Hands on Laparoscopy course appeared to contribute to expansion of laparoscopic practices. Experience gained from skills-based lectures, videotape analysis of pelvic trainer performance, and a mentored porcine laboratory resulted in most (61%) participants expanding their practice to include clinical laparoscopic suturing. PMID- 16040088 TI - Percutaneous nephrostomy catheters: drainage flow and retention strength. AB - OBJECTIVES: To evaluate the impact of percutaneous nephrostomy catheter configuration on drainage flow and retention strength. METHODS: The Cook nephrostomy 16F (symmetric balloon), Bardex Council 16F (eccentric balloon), Microvasive Flexima 14F (pigtail), and Bardex Malecot 16F (flange) nephrostomy catheters were attached to an artificial renal pelvis (12-in.-round latex balloon). The balloon was subsequently filled with either 60 mL of water or orange juice with pulp, and gravity drainage of this fluid was recorded as flow into a flowmeter. Using a Force Five Model FDV-100 force gauge, the retention strength was tested by measuring the force required to pull the nephrostomy catheter through an 8-mm hole in a 35-mm-thick biologic tissue specimen (bologna). RESULTS: The maximal flow rate using both orange juice and saline was significantly greater for the Cook nephrostomy than for the Microvasive Flexima, Bardex Malecot, and Bardex Council catheters (P < or = 0.016). The average flow rate using saline for the Cook nephrostomy catheter was significantly greater than for all other catheters (P < or = 0.02) and was significantly greater than for the Microvasive Flexima and the Bardex Council catheters (P < or = 0.036) using orange juice. The retention strength was strongest for the Cook nephrostomy catheter (3.41 +/- 0.14 lb) compared with the Bardex Council (1.75 +/- 0.1), Microvasive Flexima (1.35 +/- 0.3), and Bardex Malecot (0.29 +/- 0.03) catheters. In addition, the Microvasive Flexima catheter resulted in greater maceration of the biologic tissue after forceful dislodgement. CONCLUSIONS: The results of this study have demonstrated that the Cook nephrostomy catheter combines strong drainage flow and strong retention strength during in vitro testing. Clinical evaluations of the ease of use and patient comfort are warranted. PMID- 16040089 TI - Relationship between ankle position and pelvic floor muscle activity in female stress urinary incontinence. AB - OBJECTIVES: To assess the influence of ankle position on pelvic floor muscle (PFM) activity in women with stress urinary incontinence. METHODS: A total of 39 women, ranging in age from 38 to 72 years and clinically diagnosed with stress urinary incontinence, participated in testing of PFM activity changes during various pelvic tilt angles created by horizontal, dorsiflexed, and plantar flexed ankle positions. PFM activity was measured by an intravaginal probe with surface electromyographic electrodes. An adjustable angle platform was used to set the ankle in each of the positions to create the various pelvic tilt postures. RESULTS: Significant differences were found in resting PFM activity between horizontal standing with the ankle in the neutral position and standing with the ankle in plantar flexion (P = 0.01). Patients with ankle dorsiflexion also had greater resting PFM activity than with ankle plantar flexion (P < 0.01). Subjects showed significant changes in mean maximal PFM activity when standing with the ankle dorsiflexed and horizontal or in plantar flexion. CONCLUSIONS: A standing posture that includes various ankle positions effectively facilitates PFM activity through enhanced pelvic tilt. We recommend these ankle positions as an adjunctive option combined with PFM training for stress urinary incontinence. PMID- 16040090 TI - Endoscopic incision for functional bladder neck obstruction in men: long-term outcome. AB - OBJECTIVES: To evaluate the long-term outcomes of endoscopic bladder neck incision (BNI) for functional bladder neck obstruction. Functional and/or mechanical obstruction of the bladder neck in men causes lower urinary tract symptoms. Although alpha-blockers are the initial treatment, they often fail to alleviate the symptoms. METHODS: The records of 45 men with obstructive voiding symptoms treated between 1988 and 1996 were reviewed. The symptom score, renal parameters, urine routine microscopy, urine culture and sensitivity, uroflowmetry, voiding cystourethrography, and videourodynamic results were reviewed. Patients with a peak flow rate of less than 10 mL/s, inadequate funneling of the bladder neck, more than 40 cm H2O opening pressure with a relaxed external sphincter, a postvoid residual urine volume greater than 100 mL, no neurologic defect, and a normal urethral caliber were included. They were treated with clean intermittent catheterization and alpha-blockers. Clean intermittent catheterization was stopped when the postvoid residual urine volume was less than 50 mL. BNI was performed in patients with a poor response, noncompliance, or side effects from the alpha-blockers. Postoperatively, patients were assessed by symptom score, uroflowmetry, and postvoid residual urine volume. RESULTS: Of the 45 men, 22 responded to alpha-blockers, 5 chose clean intermittent catheterization, and 18 underwent BNI. The mean patient age was 33.42 +/- 6.56 years. The mean follow-up was 96 months (range 46 to 140). After BNI, all patients showed improvement, with the symptom score improving from 26.9 to 3.6 (P < 0.001). The peak flow rate improved from 8.5 to 19.6 mL/s (P < 0.001). This improvement persisted in more than 80% of the 18 patients. Two required a second BNI and were also doing well at the last follow-up visit. CONCLUSIONS: BNI is an effective treatment for functional bladder neck obstruction refractory to alpha-blockers and provides durable results. PMID- 16040091 TI - Efficacy of continuous local anesthetic infusion for postoperative pain after radical retropubic prostatectomy. AB - OBJECTIVES: To determine whether a subfascial continuous infusion of local anesthetic in patients undergoing radical retropubic prostatectomy would result in a reduction in postoperative opioid requirements and an improvement in pain scores. METHODS: This was a prospective, double-blind, placebo-controlled, randomized trial in patients undergoing elective radical retropubic prostatectomy. A small catheter was placed subfascially at the end of surgery and attached to an elastomeric pump that administered either 0.5% bupivacaine or normal saline into the wound at a rate of 2 mL/hr until discharge on postoperative day 3. The outcomes assessed included the dosage of hydromorphone used by a patient-controlled analgesic system, a visual analog scale (VAS) for pain at rest and with activity, a VAS of nausea, and length of hospital stay. RESULTS: A total of 100 patients were successfully randomized, with all patients completing the protocol. No differences were found between the groups with regard to VAS pain at rest, VAS pain with activity, intravenous or oral analgesic consumption, or VAS nausea scores. CONCLUSIONS: Continuous subfascial infusion of local anesthetic did not result in a postoperative reduction in opioid requirements or an improvement in pain scores in patients undergoing radical retropubic prostatectomy. PMID- 16040092 TI - Sexual function-preserving cystectomy. PMID- 16040093 TI - Effect of ureteral access sheath on stone-free rates in patients undergoing ureteroscopic management of renal calculi. AB - OBJECTIVES: To evaluate the effect of ureteral access sheaths (UASs) on stone free rates (SFRs) during ureteroscopic treatment of renal calculi. Several advantages of UASs during flexible ureteroscopy have been documented. However, no study has evaluated their impact on SFRs. METHODS: We retrospectively reviewed all ureteroscopic cases for the management of renal stones performed at our Stone Center. Data were stratified according to the use or lack of use of the UAS. The groups were stratified by stone location within the kidney. Stone-free status was determined at 2 months postoperatively by either intravenous urography with tomograms or noncontrast renal computed tomography in patients with contrast allergies. RESULTS: A total of 256 ureteroscopic procedures for the removal of renal calculi were performed between 1997 and 2003 (173 with UAS and 83 without). The groups were similar in age, sex, and stone burden. Stents were placed in nearly 80% of patients. The lower renal pole represented the most common presenting location. Stone displacement with a ureteroscopic basket for efficient fragmentation was necessary in 34%. The overall SFR in the UAS group and non-UAS group was 79% and 67%, respectively (P = 0.042). The SFRs were improved for calculi in all portions of the kidney. CONCLUSIONS: In addition to facilitating ureteroscopic access, reducing costs, and lowering intrarenal pressures, the results of the current study suggest that UASs improve SFRs during the management of renal calculi. It is now our current practice to use the UAS routinely during ureteroscopic treatment of renal and upper ureteral calculi. PMID- 16040094 TI - Changes in quality of life among low-income men treated for prostate cancer. AB - OBJECTIVES: To investigate as the primary objective changes over time in general and disease-specific health-related quality of life (HRQOL) among low-income men treated for prostate cancer in a longitudinal prospective study. METHODS: Study participants were recruited from a state-funded program providing free prostate cancer treatment to impoverished men. We included men who completed telephone interviews and self-administered questionnaires at study enrollment and 6 months of follow-up. Covariates univariately associated with HRQOL change scores were included in multivariate linear regression models. All HRQOL models were controlled for age at enrollment, race, baseline HRQOL, and treatment effects at baseline and follow-up. RESULTS: Subjects with greater baseline Gleason scores experienced more negative changes in their physical health than did men with lower Gleason scores. Men with less than a high school education experienced greater improvement in their mental well-being than did men with more than a high school education. Those experiencing treatment effects at baseline displayed greater positive changes in their urinary function than did those without treatment effects at baseline. Finally, men experiencing treatment effects at follow-up were characterized by profound reductions in sexual function compared with those free of treatment effects at follow-up. CONCLUSIONS: This work provides new insights into HRQOL over time in a low-income, multiethnic group of patients with prostate cancer. PMID- 16040095 TI - Efficacy of prostatic fossa biopsy in detecting local recurrence after radical prostatectomy. AB - OBJECTIVES: To evaluate retrospectively the efficacy of prostatic fossa biopsy in detecting local recurrence of prostate cancer in men with biochemical failure after radical prostatectomy. METHODS: Between January 1997 and December 2002, 100 men without prior adjuvant therapy after radical prostatectomy underwent transrectal ultrasound (TRUS)-guided biopsy of the prostatic fossa. The TRUS findings, digital rectal examination (DRE) findings, serum total prostate specific antigen (PSA) level at TRUS, PSA velocity, and pathologic stage and Gleason score of the radical prostatectomy specimen were correlated with the biopsy results. RESULTS: Overall, 29 (29%) of the 100 men who underwent biopsy had documented local recurrence. The sensitivity and specificity of DRE to detect biopsy-proven local recurrence was 72.4% and 64.8%, respectively. The corresponding values for TRUS were 86.2% and 53.5%. None of the men with a serum PSA concentration of less than 0.5 ng/mL at biopsy who had normal results for both TRUS and DRE had a biopsy-proven local recurrence. By stepwise multivariate logistic regression analysis, abnormal TRUS findings and serum PSA concentration at biopsy were independent predictors for positive fossa biopsy results. The combination of TRUS and serum PSA concentration was the best predictive model for a positive fossa biopsy result. CONCLUSIONS: Prostatic fossa biopsy should be avoided in patients with both or either normal DRE or TRUS findings when the PSA level is less than 0.5 ng/mL. PMID- 16040096 TI - Urinary diversion in high-risk elderly patients: modified cutaneous ureterostomy or ileal conduit? AB - OBJECTIVES: To evaluate the efficacy and safety of the type of urinary diversion during radical cystectomy in high-risk elderly patients by comparing a modified cutaneous ureterostomy with the ileal conduit urinary diversion. METHODS: Of 481 patients who underwent radical cystectomy between 1993 and 2002, 54 individuals older than 75 years with an American Society of Anesthesiologists score of 3e, 4, or 4e were characterized as high risk. These patients were grouped according to those who underwent a modified cutaneous ureterostomy (group 1, 29 patients) and those who underwent ileal conduit urinary diversion (group 2, 25 patients). Student's t and Fisher's exact tests were used for statistical analysis. RESULTS: The patients in group 2 had a longer operative time (P < 0.001), an increased need for blood transfusion (P = 0.025), an increased need for intensive care monitoring (P = 0.032), and a longer mean hospitalization time (P < 0.001) than the patients in group 1. In group 2, we recorded statistically greater rates of intraoperative complications (P = 0.035), early medical and surgical complications (P = 0.031 and P = 0.012, respectively), and late surgical complications (P = 0.004). The intraoperative, early, and late surgical complication rate was 13.7%, 24.1%, and 17.2% in group 1 and 40%, 60%, and 56% in group 2, respectively. One patient in group 2 died in the early postoperative period. CONCLUSIONS: Cutaneous ureterostomy represents a simplified alternative for urinary diversion in high-risk elderly patients. It can be performed quickly, with few early and late postoperative complications compared with the ileal conduit operation. PMID- 16040097 TI - Loss of coxsackie and adenovirus receptor expression is associated with features of aggressive bladder cancer. AB - OBJECTIVES: To examine whether loss of coxsackie and adenovirus receptor (CAR) expression is associated with bladder cancer characteristics and clinical outcomes and with expression of p53 and E-cadherin. Low levels of CAR are associated with decreased efficiency of adenovirus-mediated gene transduction of bladder transitional cell carcinoma. METHODS: Immunohistochemical staining for CAR and p53 was carried out on tissue microarrays from 62 patients who had undergone radical cystectomy. We also examined 30 specimens for E-cadherin expression. RESULTS: CAR expression was lost in 17 (27%) of 62 tumors. Loss of CAR expression was associated with metastases to regional lymph nodes (P = 0.049), muscle-invasive disease (P = 0.025), grade 3 disease (P = 0.038), altered p53 status (P = 0.041), and loss of E-cadherin expression (P = 0.042). With a median follow-up of 60 months, loss of CAR expression was associated with decreased bladder cancer-specific survival (P = 0.029) but not disease progression. When adjusted for the effects of standard pathologic features, only lymph node metastasis was associated with bladder transitional cell carcinoma progression and mortality. CONCLUSIONS: Loss of CAR expression is associated with established markers of biologically aggressive bladder transitional cell carcinoma. The association of CAR with E-cadherin and p53 suggests a potential role for CAR in the regulation of urothelium integrity and the cell cycle. PMID- 16040098 TI - Malignant mesothelioma of the tunica vaginalis. AB - OBJECTIVES: To review our experience with the management of malignant mesothelioma of the tunica vaginalis with emphasis on disease-related outcomes. METHODS: A retrospective chart review of patients seen during the past 25 years at our cancer center identified 5 cases of malignant mesothelioma of the tunica vaginalis. RESULTS: The mean age of patients at presentation was 61.2 years (range 57 to 83). Asbestos exposure was identified in 4 patients. Three patients presented with clinical symptoms suggestive of a hydrocele and two presented with clinical signs of an inguinal hernia. The final diagnosis was established intraoperatively in 1 patient and postoperatively in the remaining 4. Radical orchiectomy or hernia sac with spermatic cord excision was the primary treatment modality. Although radical surgical treatment achieved negative resection margins in 4 cases, 4 of 5 patients died of the disease, with a mean disease-specific survival of only 29 months (range 5 to 68). Regional inguinal lymph node metastasis developed in 3 of 5 patients. Salvage therapy did not prove curative in the 2 patients who received it. CONCLUSIONS: Malignant mesothelioma of the tunica vaginalis constitutes a rare but often fatal malignancy of the male genitalia. This diagnosis should be suspected in patients exposed to asbestos and presenting with clinical symptoms of either hydrocele or inguinal hernia. Frequent inguinal lymph node involvement indicates a potential role of inguinal lymphadenectomy in the primary treatment. PMID- 16040099 TI - Bladder exstrophy in the newborn: a snapshot of contemporary practice patterns. AB - OBJECTIVES: To use a large nationwide database to investigate and describe practice patterns in the contemporary management of bladder exstrophy. METHODS: The Healthcare Cost and Utilization Project Nationwide Inpatient Sample (1988 to 2000) was queried to identify infants with bladder exstrophy hospitalized during the first week after birth. Admission and disposition patterns, resource utilization and length of stay, surgical repair trends, and factors associated with in-hospital death were analyzed. RESULTS: We identified 426 hospital admissions of newborns with exstrophy. Most patients (75%) were transferred in from, or out to, other facilities; this was a fundamental feature of early exstrophy care. Racial differences were evident, with Hispanics less likely to be transferred (19% versus 60%, P = 0.001). Among newborns who were not transferred, many (46%) were discharged without bladder surgery. Surgical repair was usually done in a hospital other than the birth hospital; the mean hospital charges for surgery were 75,742 dollars. Of the 5 patients who died after repair, all had undergone surgery at "low-volume" hospitals. The length of stay did not change significantly during the study period, helping to keep resource utilization high in this population. CONCLUSIONS: The results of this study provide a "snapshot" of bladder exstrophy practice patterns during the newborn period between 1988 and 2000. Additional research should investigate whether newborns with exstrophy are receiving optimal care, including appropriate timing of surgery, equitable transfers to tertiary centers, and reconstruction at centers with adequate volume and experience. PMID- 16040100 TI - Modified clipless antegrade nerve preservation in robotic-assisted laparoscopic radical prostatectomy with validated sexual function evaluation. AB - INTRODUCTION: We present our technique for clipless antegrade neurovascular bundle preservation during robotic laparoscopic radical prostatectomy, along with short-term follow-up of our patients' sexual function. TECHNICAL CONSIDERATIONS: Using the da Vinci three-arm robotic system, we performed robotic laparoscopic radical prostatectomy using a transperitoneal approach in an antegrade fashion. After division of the bladder neck, the posterior plane of the prostate was developed distally toward the apex of the prostate in the midline. This plane was then developed completely, releasing the vascular pedicles and neurovascular bundles in a medial to lateral direction, with occasional use of bipolar cautery and without the use of clips or monopolar cautery. Patients with a minimal follow up of 3 months who did not require open conversion were included in this study. A total of 56 patients met these inclusion criteria between February 2003 and May 2004, with a mean follow-up of 6 months. Patients were given the validated Rand Medical Outcomes Study 36-Item Health Survey, version 2, with the University of California, Los Angeles, Prostate Cancer Index preoperatively and at 1, 3, 6, and 12 months postoperatively. The overall score for both unilateral and bilateral nerve-sparing groups was 35.0, 39.7, 49.4, and 49.6 at 1, 3, 6, and 12 months postoperatively, respectively. These coincided with a return to baseline potency rate of 47%, 54%, 66%, and 69% at 1, 3, 6, and 12 months, respectively. CONCLUSIONS: Antegrade dissection of the neurovascular bundle, avoiding the use of clips or monopolar cautery during robotic laparoscopic radical prostatectomy, may result in early return of sexual function and overall outcome similar to that after radical retropubic prostatectomy. PMID- 16040101 TI - Evaluation of surgical models for renal collecting system closure during laparoscopic partial nephrectomy. AB - OBJECTIVES: To evaluate the utility of the porcine and canine models for collecting system closure after partial nephrectomy involving violation of the renal collecting system. Advanced surgical technologies and novel techniques for performing laparoscopic partial nephrectomy are frequently evaluated in an animal model. METHODS: After evaluation of the upper urinary tract with retrograde pyelography, laparoscopic partial nephrectomy, including violation of the renal collecting system, was performed in 8 domestic pigs (group 1) and 2 dogs (group 2) with documented normal upper urinary tract physiology. No collecting system closure was performed. Subsequently, in an attempt to generate an adequate model for renal collecting system closure, 6 pigs (group 3) underwent partial obstruction of the proximal ureter before laparoscopic partial nephrectomy without collecting system closure. After 3 weeks, the animals underwent evaluation, including retrograde pyelography, and were killed for histopathologic evaluation of the upper urinary tracts. RESULTS: Despite documented transection of major calices in all animals, none with physiologically normal upper urinary tracts developed urinomas. After partial occlusion of the proximal ureter, all pigs had hydronephrosis, but only one of six developed a urinoma. CONCLUSIONS: The porcine and canine models are inadequate for evaluating renal collecting system closure after partial nephrectomy. Our high-pressure partial occlusion model was similarly inadequate. The limitations of these animal models should be considered when reviewing published data regarding technologies for partial nephrectomy. Future data regarding novel technologies for partial nephrectomy should also be carefully considered before clinical application. PMID- 16040102 TI - Comprehensive urodynamics evaluation of 146 men with incontinence after radical prostatectomy. AB - OBJECTIVES: To assess the filling, storage, and voiding urodynamic parameters in a large group of men with urinary incontinence after radical prostatectomy. METHODS: We reviewed the videourodynamics testing results for 146 consecutive men referred for urinary incontinence after radical prostatectomy. RESULTS: The mean patient age was 69.0 years (range 48 to 85), and the mean interval since radical prostatectomy was 4 years (range 4 months to 19 years). All but four tests were performed more than 12 months postoperatively. Stress urinary incontinence (SUI) was demonstrated in 139 men (95%), with a mean abdominal leak point pressure of 59 cm H2O. A statistically significant correlation was found between the leak point pressure and static urethral pressure profilometry measurements (r = 0.46, P < 0.0001). The mean urethral pressure profilometry measurements in those with SUI were significantly lower than in those without (46.6 versus 69 cm H2O, P = 0.001). A total of 34 patients had diminished compliance or detrusor instability, but this was the sole finding in only 3. A hypocontractile detrusor response was seen in 49 patients, and 35 of these augmented voiding by abdominal straining. Patients with previous radiotherapy (n = 24) were more likely to have bladder outlet obstruction; the other parameters were similar to those in patients without radiotherapy. CONCLUSIONS: Incontinence after radical prostatectomy is caused by intrinsic sphincter deficiency in the vast majority of patients. Urethral pressure profilometry measurements correlated with the severity of SUI, as measured by abdominal leak point pressure. Bladder outlet obstruction may coexist with SUI in a significant portion of patients. During voiding, a hypocontractile detrusor response may be seen, but the clinical significance of this finding is unclear. PMID- 16040103 TI - Health-related quality-of-life outcomes after anatomic retropubic radical prostatectomy in the phosphodiesterase type 5 ERA: impact of neurovascular bundle preservation. AB - OBJECTIVES: To examine the impact of neurovascular bundle preservation on longitudinal health-related quality-of-life (HRQOL) outcomes after anatomic radical retropubic prostatectomy (RP) using a validated questionnaire. METHODS: We examined patient-reported sexual and urinary HRQOL at baseline and at 3, 6, 12, and 24 months after RP using the University of California, Los Angeles, Prostate Cancer Index among 342 patients treated between 2001 and 2004 by a single surgeon. The time to return to baseline urinary and sexual function and bother were compared between men who underwent unilateral versus bilateral nerve sparing RP using a Cox proportional hazards regression model. RESULTS: Of the 342 patients, 15 (5%), 69 (20%), and 258 (75%) had no, one, or both neurovascular bundles preserved, respectively. After adjustment for age and baseline sexual function, bilateral nerve sparing was associated with greater sexual function scores than unilateral nerve sparing at all points, although the differences only approached or reached significance at 3 (P = 0.06) and 6 (P = 0.04) months after RP. After adjustment for age and baseline sexual function, a trend was noted for an earlier return to baseline sexual function among men who underwent bilateral nerve-sparing RP (hazard ratio 1.67, 95% confidence interval 0.88 to 3.17, P = 0.12), although this did not reach significance. More than 90% of the men returned to their baseline urinary function and bother, regardless of nerve sparing status. CONCLUSIONS: In the current study, bilateral nerve-sparing RP was associated with better postoperative sexual HRQOL scores than unilateral nerve sparing RP, although in general the differences were slight. PMID- 16040104 TI - Treatment of testicular cancer: influence on pituitary-gonadal axis and sexual function. AB - OBJECTIVES: To investigate the influence of treatment for testicular cancer on the pituitary-gonadal axis and sexual function in long-time survivors after unilateral orchiectomy. METHODS: Blood was drawn from patients treated for testicular cancer during routine oncologic follow-up for measurement of luteinizing hormone, follicle-stimulating hormone, sexual hormone-binding globulin, testosterone, and bioavailable testosterone. Sexual function was evaluated using the International Index of Erectile Function 15-item (IIEF-15) questionnaire. Patients were grouped according to treatment: group 1 followed a surveillance strategy, group 2 received two cycles of carboplatin monotherapy, and group 3 underwent cisplatin, etoposide, and bleomycin chemotherapy. RESULTS: No statistically significant difference was found in the serum hormonal levels among the three groups, and all hormonal levels were within the 95% confidence range, except for follicle-stimulating hormone. The median serum testosterone level was 3.5 ng/mL in group 1, 3.9 ng/mL in group 2, and 4.2 ng/mL in group 3. In group 1, the median IIEF-15 score was 64.0, and the median Erectile Function (EF) domain score was 28. The median scores in groups 2 and 3 were 62.5 for IIEF 15 and 27.5 for EF and 65.0 for IIEF-15 and 30.0 for EF, respectively. No correlation was found between testosterone level and IIEF-15 or EF score. CONCLUSIONS: None of the treatments investigated had a significant influence on the serum hormonal levels in long-time survivors of testicular cancer. Patients undergoing chemotherapy have no greater risk of developing a hormonal disorder than those following a surveillance strategy, and therapy for testicular cancer is not a risk factor for erectile dysfunction. PMID- 16040105 TI - Intravesical interleukin-12 gene therapy in an orthotopic bladder cancer model. AB - OBJECTIVES: To evaluate the antitumor effect of intravesical cationic liposome mediated interleukin-12 (IL-12) gene delivery in an orthotopic murine bladder cancer model, and to investigate the immunologic memory against tumors between IL 12 gene therapy and bacille Calmette-Guerin (BCG) therapy. METHODS: Orthotopic murine bladder tumors were established by implanting 5 x 10(5) MBT-2 cells into the bladder of syngeneic female C3H mice. Intravesical IL-12 gene therapy was evaluated at varying doses: 0 microg (control) and 3, 5, and 10 microg (n = 8 for each group). Intravesical treatments were performed every 3 days and repeated six times beginning 5 days after tumor implantation. To compare the long-term, tumor specific immunity between IL-12-treated mice (n = 18) and BCG-treated mice (n = 20), the animals surviving at day 60 and 10 new control mice were rechallenged with MBT-2 cells and received no additional treatment. On day 120, all surviving mice were killed and underwent necropsy. RESULTS: In the IL-12 groups at doses of 0, 3, 5, and 10 microg, 0, 2, 3, and 3 mice survived, respectively. Mice in the 5 microg and 10-microg IL-12 groups survived significantly longer than did the control group. All mice cured by IL-12 treatment successfully rejected the rechallenge with MBT-2 cells; however, mice cured by BCG and the new control mice died of the rechallenged bladder tumors. CONCLUSIONS: Intravesical IL-12 gene therapy, which induced long-lasting tumor-specific immunologic memory compared with BCG therapy, improved survival in an orthotopic bladder cancer model. PMID- 16040106 TI - Potential antidepressant-like effect of MTEP, a potent and highly selective mGluR5 antagonist. AB - The involvement of glutamate in the pathophysiology of depression has been suggested by a number of experiments. It was well established that compounds, which decreased glutamatergic transmission via blockade of NMDA receptor, produced antidepressant-like action in animal tests and models. The present study was carried out to investigate whether a selective mGluR5 antagonist 3-[(2-methyl 1,3-thiazol-4-yl)ethynyl]-pyridine (MTEP) induces antidepressant-like effects after intraperitoneal injections in male Wistar rats or male C57BL/6J mice. Potential antidepressant-like activity of MTEP was evaluated using the forced swimming test (FST) in rats, the tail suspension test (TST) in mice and the olfactory bulbectomy (OB) model of depression in rats. The results of our studies showed, that MTEP (0.3-3 mg/kg) produced a significant dose-dependent decrease in the immobility time of mice in the TST, however, at doses of 1 or 10 mg/kg, it did not influence the behavior of rats in the FST in rats. Moreover, the repeated administration of MTEP (1 mg/kg) attenuated the OB-related hyperactivity of rats in the open field test, in the manner similar to that seen following chronic (but not acute) treatment with typical antidepressant drugs. These data suggest that MTEP, which is considered to be a potential therapeutic agent, may play a role in the therapy of depression. PMID- 16040107 TI - On the role of schistosome mating structure in the maintenance of drug resistant strains. AB - The effects of drug treatment of human hosts upon a population of schistosome parasites depend upon a variety of factors. Previous models have shown that multiple strains of drug-resistant parasites are likely to be favored as the treatment rate increases. However, such models have neglected to account for the complex nature of schistosome mating biology. To more accurately account for the biology of these parasites, a simple mating structure is included in a multi strain schistosome model, with parasites under the influence of drug treatment of their human hosts. Parasites are assumed to pay a cost for drug resistance in terms of reduced reproduction and transmission. The dynamics of the parasite population are described by a system of homogeneous differential equations, and the existence and stability of the exponential solutions for this system are used to infer the impact of drug treatment on the maintenance of schistosome genetic diversity. PMID- 16040109 TI - Generalizations on variations in comprehension and production: a further source of variation and a possible account. PMID- 16040108 TI - Neural modeling and imaging of the cortical interactions underlying syllable production. AB - This paper describes a neural model of speech acquisition and production that accounts for a wide range of acoustic, kinematic, and neuroimaging data concerning the control of speech movements. The model is a neural network whose components correspond to regions of the cerebral cortex and cerebellum, including premotor, motor, auditory, and somatosensory cortical areas. Computer simulations of the model verify its ability to account for compensation to lip and jaw perturbations during speech. Specific anatomical locations of the model's components are estimated, and these estimates are used to simulate fMRI experiments of simple syllable production. PMID- 16040110 TI - Do differences in brain activation challenge universal theories of dyslexia? AB - It has been commonly agreed that developmental dyslexia in different languages has a common biological origin: a dysfunction of left posterior temporal brain regions dealing with phonological processes. Siok, Perfetti, Jin, and Tan (2004, Nature, 431, 71-76) challenge this biological unity theory of dyslexia: Chinese dyslexics show no deficits in posterior temporal brain regions but a functional disruption of the left middle frontal gyrus. Here, I will argue that these data do not challenge universal cognitive theories of dyslexia according to which weaknesses in the ability to process the phonological features of language are at the origin of dyslexia. PMID- 16040111 TI - Minimization of the inevitable residual monomer in denture base acrylic. AB - OBJECTIVES: Residual monomer ([MMA](R)) in denture base acrylic continues to be of concern. The response surface of concentration vs. time and temperature for the equilibration of methyl methacrylate (MMA) and its polymer (PMMA) allows a prediction of the time to the minimum at any temperature for a closed system. It was the purpose here to determine whether this prediction applies to normal denture base processing, and whether optimum conditions could be identified. METHODS: Denture bases were processed following normal laboratory procedures, including pre-cure for 3 h at 70 degrees C for all tests. Commercial powder and liquid were used at either 95 or 100 degrees C, or a plain PMMA powder and the same liquid at 95 degrees C, for times ranging from 5 to 192 h. Residual MMA was determined by gas chromatography. RESULTS: [MMA](R) decreased steadily from approximately 0.25% to as low as approximately 0.07% with increasing time at temperature, but did not approach equilibrium. The rate of diffusive loss of MMA appears to exceed the rate of depolymerization. SIGNIFICANCE: Residual monomer is inevitable for all PMMA-based products no matter what the curing conditions are. However, extended time at high temperature can allow low values to be attained, and the time allowed can compensate for processing temperatures somewhat lower than the ordinarily recommended 100 degrees C. It is suggested that overnight processing at 95 degrees C should be adopted to minimize [MMA](R) and save energy. This result is of importance for work at high altitude. PMID- 16040112 TI - Influence of the application time on the penetration of different dental adhesives and a fissure sealant into artificial subsurface lesions in bovine enamel. AB - Sealing of approximal enamel lesions by infiltration with low viscous resins seems to be a promising approach in non-operative dentistry and should bear advantages compared to remineralization or invasive treatment. OBJECTIVE: The aim of this in vitro study was to evaluate the penetration ability of five dental adhesives and a fissure sealant into initial enamel lesions for an application time of either 15 s or 30 s. METHODS: In each of 54 specimens of bovine enamel, three windows were demineralized for 14 days. Subsequently, two windows were etched with phosphoric acid for 5s in order to degrade the surface layer, whereas one window served as untreated control. The specimens were randomly divided into six groups and a fissure sealant as well as five different adhesives were applied onto the subsurface lesions and allowed to penetrate for either 15 or 30 s. Overlying material was wiped away and the resins were light cured. To visualize the penetrated resins and the remaining pore structures, the specimens were infiltrated with a low viscous fluorescent resin and studied using a Confocal Laser Scanning Microscope (CLSM). RESULTS: For Helioseal, Heliobond, Resulcin Monobond, and Excite an application time of 30s resulted in significantly higher (p < 0.05; t-test) penetration depths (47-105 microm) compared to 15s (29-49 microm). SIGNIFICANCE: Helioseal, Heliobond, Resulcin Monobond, and Excite are suitable for sealing artificial subsurface enamel lesions in vitro. After an application time of 30 s a significant deeper penetration could be observed for these materials. PMID- 16040113 TI - Strength and fracture pattern of monolithic CAD/CAM-generated posterior crowns. AB - OBJECTIVES: This study evaluated the strength and fracture pattern of monolithic posterior CAD/CAM crowns hypothesizing that zinc-phosphate cemented lithium disilicate crowns might show the same fracture strength as adhesively cemented crowns. METHODS: Two sets of monolithic posterior crowns each with uniform occlusal and lateral wall thickness of 1.5mm were fabricated from three types of block ceramic (1) lithium disilicate glass, (2) leucite glass and (3) feldspathic ceramic using CEREC 3 CAD/CAM. Crowns (n = 15) of ceramics (1), (2) and (3) each were (A) zinc-phosphate cemented, (B) adhesively cemented on resin-based composite dies and loaded until fracture. Load data was analyzed using ANOVA and Scheffe tests. Crack pattern was evaluated on an additional three sample cross sections for each group at start of fracture. RESULTS: Radial cracks originated early at the cementation interfaces and cone cracks were observed finally at the loading sites. Mean load values (SD) of A-crowns at fracture start/end (1) 807 (91) N/2082 (192) N; (2) 915 (193) N/1130 (166) N; (3) 985 (199) N/1270 (301) N were all significantly (P < 0.001) lower when compared to their B-crown analogs (1) 1456 (205) N/2389 (84) N; (2) 1684 (395) N/2469 (171) N; (3) 1548 (304) N/2392 (75) N, rejecting the authors hypothesis. A-1 crowns had significantly (P < 0.001) higher fracture load than A-2 and A-3 crowns. The A-1 crown fracture load data, even if significantly (P < 0.001) lower, came close to the B-1 values. SIGNIFICANCE: Adhesive cementation balanced the strength of weak ceramics with that of strong ceramic and recommended itself for leucite glass ceramic and feldspathic ceramic crowns. Zinc-phosphate cementation appeared feasible for lithium disilicate crowns. PMID- 16040114 TI - Micro-tensile bond strength of three luting resins to human regional dentin. AB - OBJECTIVES: To evaluate the micro-tensile bonding strength (muTBS) of three luting resins to human regional dentin. METHODS: Dentin disks from non-carious third molars were prepared from different regions (s, superficial dentin; d, deep dentin; c, cervical dentin), and divided into groups based on anatomical locations and luting resins (Super-Bond C&B: SB; Panavia F 2.0: PF; RelyX Unicem: RU): SB-s, SB-d, SB-c; PF-s, PF-d, PF-c; RU-s, RU-d, RU-c. Luting resins were used according to the manufacturers' instructions, to bond 1-mm-diameter PMMA or composite rods to the exposed dentin specimens under a load of 7.5 N, in the self curing mode. After storage for 1 or 3 days, muTBS was tested at a cross-head speed of 1mm/min. Data were analyzed with ANOVA and Fisher's PLSD test. The bonding interface and fractography analyses were performed with SEM and TEM. RESULTS: ANOVA results showed that muTBS to superficial dentin was significantly higher than to deep or cervical dentin for all three luting resins. SB-s and PF s, with the highest muTBS, failed primarily cohesively in luting resin. muTBS of SB-d and SB-c were significantly higher than those of PF and RU. RU, with the lowest regional muTBS, failed mostly within demineralized dentin. SEM and TEM showed that adhesive failures in SB and PF occurred at the top of the hybrid layer (HL), but no obvious HL was observed in RU. SIGNIFICANCE: Luting resins with different chemical formulations and applications yield significantly different bond strengths to different regions in human dentin. PMID- 16040115 TI - Low shrinkage light curable nanocomposite for dental restorative material. AB - OBJECTIVES: The aim of this study was to develop a low shrinkage visible light curable nanocomposite dental restorative material without sacrificing the other properties of conventional materials. This nanocomposite was developed by using an epoxy resin 3,4-epoxycyclohexylmethyl-(3,4-epoxy)cyclohexane carboxylate (ERL4221) matrix with 55% wt of 70-100 nm nanosilica fillers through ring-opening polymerization. GPS (gamma-glycidoxypropyl trimethoxysilane) was used to modify the surfaces of silica nanoparticles. RESULTS: The nanocomposite was shown to exhibit low polymerization shrinkage strain, which is only a quarter of currently used methacrylate-based composites. It also exhibited a low thermal expansion coefficient of 49.8 microm/m degrees C which is comparable to that of the methacrylate based composites (51.2 microm/m degrees C). The strong interfacial interactions between the resin and fillers at nanoscales were demonstrated by an observed high strength and high thermal stability of the nanocomposite. A microhardness of 62 KHN and a tensile strength of 47 MPa were reached. A high degree of conversion ( approximately 70%) can be obtained after less than 60 s of irradiation upon the nanocomposite. A transmission electron microscope (TEM) study of the nanocomposite showed no aggregation of fillers. Comparable results to the methacrylate based composites were obtained from the one day MTT (3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) cytotoxicity test. SIGNIFICANCE: The developed epoxy resin based nanocomposite demonstrated low shrinkage and high strength and is suitable for dental restorative material applications. PMID- 16040116 TI - Roles of nitric oxide and prostacyclin in triethyleneglycol dimethacrylate (TEGDMA)-induced vasorelaxation. AB - OBJECTIVES: Most dental resinous materials contain the diluent monomer triethyleneglycol dimethacrylate (TEGDMA), which has been reported to be bioactive. Previously, it was demonstrated that TEGDMA induces vasorelaxation. The present study examines the mechanism(s) of the TEGDMA-induced vasorelaxation by measuring vascular nitrite and prostacyclin levels. METHODS: Nitrite and prostacyclin levels were assayed in rat aortic tissues in response to TEGDMA. The involvement of guanylyl and adenylyl cyclases in TEGDMA-induced aortic vasorelaxation was determined using the enzyme inhibitors 1H [1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) and 9-(tetrahydro-2-furanyl)-9H purin-6-amine (SQ22536), respectively. RESULTS: TEGDMA enhanced the levels of nitrites in endothelium-intact and that of protacyclin in both endothelium-intact and denuded rat aortas. The increase in nitrites was associated with endothelium dependent aortic relaxation mediated via the activation of guanylyl cyclase, while the increase in prostacyclin was associated with both endothelium-dependent and independent relaxation linked to adenylyl cyclase stimulation. SIGNIFICANCE: Data from the present investigation can be relevant to dental practice employing materials containing TEGDMA by providing insights into the vasorelaxant effect of the monomer following placement of the materials in the oral cavity. Additional studies that are more relevant to the clinical situation are required to confirm these initial results and further explore their implications. PMID- 16040117 TI - Qualitative computer aided evaluation of dental impressions in vivo. AB - OBJECTIVES: Clinical investigations dealing with the precision of different impression techniques are rare. Objective of the present study was to develop and evaluate a procedure for the qualitative analysis of the three-dimensional impression precision based on an established in-vitro procedure. The zero hypothesis to be tested was that the precision of impressions does not differ depending on the impression technique used (single-step, monophase and two-step techniques) and on clinical variables. METHODS: Digital surface data of patient's teeth prepared for crowns were gathered from standardized manufactured master casts after impressions with three different techniques were taken in a randomized order. Data-sets were analyzed for each patient in comparison with the one-step impression chosen as the reference. The qualitative analysis was limited to data-points within the 99.5%-range. Based on the color-coded representation areas with maximum deviations were determined (preparation margin and the mantle and occlusal surface). To qualitatively analyze the precision of the impression techniques, the hypothesis was tested in linear models for repeated measures factors (p < 0.05). RESULTS: For the positive 99.5% deviations no variables with significant influence were determined in the statistical analysis. In contrast, the impression technique and the position of the preparation margin significantly influenced the negative 99.5% deviations. SIGNIFICANCE: The influence of clinical parameter on the deviations between impression techniques can be determined reliably using the 99.5 percentile of the deviations. An analysis regarding the areas with maximum deviations showed high clinical relevance. The preparation margin was pointed out as the weak spot of impression taking. PMID- 16040118 TI - Polymerization shrinkage and contraction stress of dental resin composites. AB - OBJECTIVE: The aim of this study was to evaluate the shrinkage, contraction stress, tensile modulus, and the flow factor of 17 commercially available dental resin composites. METHOD: The volumetric shrinkage measurements were performed by mercury dilatometry, and the contraction stress and tensile modulus were determined by means of stress-strain analysis. The statistical analysis was conducted by ANOVA and Tukey's post hoc test, and linear regression. RESULTS: Strong linear correlation for most resin composites were found for (i) contraction stress and shrinkage (ii) contraction stress and tensile modulus, and (iii) shrinkage and tensile modules. For most of the materials the unpolymerized resin content determines the amount of shrinkage, contraction stress and tensile modules. The pre-polymerized clusters in Heliomolar results in improved shrinkage/contraction stress properties. The shrinkage/contraction stress for Filtek Z100, Aelite Flo, and Flow-it was too high for the amount of resin in the resin composite. This was rationalized by high polymerization rates, a flow factor, and the nature of the resin. SIGNIFICANCE: High shrinkage and/or high contraction stress may lead to failure of the bond between the resin composites and the tooth structure. This study shows that the unpolymerized resin content determines the amount of shrinkage, contraction stress and tensile modules. Therefore, using pre-polymerized clusters will improve shrinkage/contraction stress properties, as was shown in Heliomolar, while high polymerization rates, and low flow factors have a deteriorative effect on the shrinkage/contraction stress properties. PMID- 16040119 TI - Hypothesis paper Brain talks with fat--evidence for a hypothalamic-pituitary adipose axis? AB - The adipose tissue signals to the brain via its secretory products. However, it is unknown whether the brain itself can directly contact the fat tissue. In order to test this hypothesis, the adipocytic expression of receptors for pituitary hormones and hypothalamic peptides was investigated. Besides FSH- and LH receptors, adipocytes do express the specific receptors for ACTH, TSH, GH, prolactin, oxytocin and the three receptor subtypes for vasopressin. Thus, the adipose tissue might no longer be regarded as an inert and steady tissue but as a fast acting player downstream of and under the control of the brain. Based on this, the potential existence and clinical impact of a hypothalamic-pituitary adipose axis should further be investigated. PMID- 16040120 TI - Insights into the genetic organization of the Bacillus mycoides cryptic plasmids pDx14.2 and pSin9.7 deduced from their complete nucleotide sequence. AB - Bacillus mycoides, a member of the Bacillus cereus group of bacteria, can be easily distinguished from close species because of colony shape, made by filaments of cells, resembling fungal hyphae, curving clock- or counterclockwise depending on the strain. Two plasmids, one from a strain curving to the right (pDx14.2), the other from a strain curving to the left (pSin9.7), were sequenced and analyzed for gene content and replication mode. Rolling-circle replication modules and mobilization proteins were found, very similar to those of other plasmids of the B. cereus group bacilli, mostly Bacillus thuringiensis living in the same ecosystem, suggesting active plasmid exchange in nature. PMID- 16040121 TI - The bioconcentration factor of perfluorooctane sulfonate is significantly larger than that of perfluorooctanoate in wild turtles (Trachemys scripta elegans and Chinemys reevesii): an Ai river ecological study in Japan. AB - Turtles rank high in the river food chain, and are suitable for predicting the bioconcentrations of chemicals through the food chain. Trachemys scripta elegans (N=46) and Chinemys reevesii (N=51) were captured in a river in Japan, from September to October 2003 and April to June 2004. Surface water samples were collected simultaneously from the same sites at which the turtles were caught. Serum perfluorooctane sulfonate (PFOS) ranged from 2.4 to 486 microg/L, while water PFOS levels ranged from 2.9 to 37 ng/L. The geometric mean (GM) (geometric standard deviation, GSD) of the bioconcentration factor (BCF) of PFOS was 10,964 (2.5). In contrast, the perfluorooctanoate (PFOA) level in water ranged from 16.7 87,100 ng/L, and serum PFOA ranged from <0.2 to 870 microg/L. The GM (GSD) of the BCF of PFOA was 3.2 (7.9). Furthermore, the BCF of PFOA decreased as the PFOA level in the surface water increased. PFOS could be preferentially bioconcentrated in biota, and PFOA, slightly bioconcentrated. PMID- 16040122 TI - The relative sensitivity of growth and reproduction in the springtail, Folsomia candida, exposed to xenobiotics in the laboratory: an indicator of soil toxicity. AB - The Folsomia candida reproduction test [ISO, 1998. Soil quality--Inhibition of reproduction of Collembola (Folsomia candida) by soil pollutants. International Standard Organization Report 11267, 1998, Geneva] is used to evaluate the ecotoxicological risks of contaminants in soils. The aim of this study was to compare the sensitivity of growth and reproduction of F. candida to four xenobiotics: two metals (Cd, Al), one metalloid (As), and one organic compound (pentachlorophenol). We showed that reproduction is a slightly more sensitive parameter than growth: EC(20) for reproduction was 1.25 microg/g dry soil for arsenic, 56 microg/g for cadmium, 97.5 microg/g for aluminum, and 41.7 microg/g for pentachlorophenol. The corresponding EC(20) values for growth were 2.8, 65, 630, and 94.6 microg/g. Keeping in mind that a growth test needs fewer juveniles and less time than a reproduction test, we conclude that the two parameters are complementary and could be used for a better ecotoxicological evaluation of contaminants. However, the relative growth and reproduction sensitivities should be tested with more chemicals before growth could be considered as a good alternative for a faster sublethal test. PMID- 16040123 TI - Variation of POP concentrations in fresh-fallen snow and air on an Alpine glacier (Monte Rosa). AB - To understand better the mechanisms ruling the fate of POPs (persistent organic pollutants) in cold environments, a field campaign sampling fresh-fallen snow and air on an Alpine glacier was carried out during Summer 2003. The concentrations of all analyzed chemicals in fresh-fallen snow show a sharp decrease over time, particularly for the more volatile POPs, confirming the rather limited literature evidence of a rapid decline of such substances from the snowpack and/or ice. Even if the results presented here are preliminary and should be confirmed by further studies, some evidence of the influence of a night/day cycle of temperature on POP deposition and revolatilization mechanisms has been highlighted. Finally, the role of cold condensation and long-range atmospheric transport in the contamination of higher altitudes in this Alpine system has been substantiated, particularly for OC pesticides. PMID- 16040124 TI - Coagulant toxicity and effectiveness in a slaughterhouse wastewater treatment plant. AB - Attempts were made in this study to examine the toxicity of polymer/alum addition to the aeration tank effluent prior to sludge flotation as practiced in a slaughterhouse wastewater treatment plant. Based on the Microtox toxicity assay, alum at concentrations 100-200 mg/L was found to slightly increase the toxicity level of slaughterhouse wastewater effluent. However, at higher concentrations (300-1000 mg/L), significant residual chronic toxicity remained in all slaughterhouse wastewater effluents, independent of the treatment process. Polymer, on the other hand, removed organics and solids, but polymer effluents are more toxic than alum at extremely low concentration. Results indicated that alum and polymer caused inhibitory effects to the system at soluble concentrations of approximately 400 and 60 mg/L and above, respectively. The data also indicated that the solids collected in both tests (polymer/alum) were much more toxic than those from the effluents. Sediment samples from the polymer tests were the most toxic. Furthermore, the effluent toxicity of the coagulants was dramatically more significant when used after the settlement of solids than when used in mixed liquor. In addition, strong correlations were observed between the observed toxicity for a series of supernatants and the coagulant concentrations of alum/polymer processes, and between supernatants and solids collected in both tests. PMID- 16040125 TI - Temporal stability of symptom dimensions in adult patients with obsessive compulsive disorder. AB - BACKGROUND: Although symptoms of obsessive-compulsive disorder (OCD) are heterogeneous, considerable advances have been made in subtyping OCD based on factor-analysed symptom dimensions. However, there is very little empirical data on the longitudinal course of symptom dimensions in adult OCD. METHODS: We examined prospectively the temporal symptom stability in adult OCD patients. Of 54 baseline OCD inpatients, 43 (80%) were re-assessed with the Yale-Brown Obsessive-Compulsive Scale symptom checklist after 6 years on average. RESULTS: Significant changes occurred within the symptom dimensions aggressive/checking, symmetry/ordering, and contamination/cleaning, whereas the others (hoarding, sexual/religious) remained unchanged from baseline to follow-up. Shifts between different dimensions from baseline to follow-up were rare, the score of each dimension at follow-up was most strongly predicted from the score of the same dimension at baseline. LIMITATIONS: The main limitation of the present study is the relatively small sample size. Furthermore, not the same raters conducted the baseline and follow-up assessments. CONCLUSIONS: Symptom dimensions seem to be remarkably stable over several years in adult OCD, despite various treatments and significant improvements in symptom severity. This underlines the usefulness of these symptom dimensions for studies of biological and genetic markers, comorbidity and treatment response predictors. PMID- 16040126 TI - The Calgary Depression Rating Scale for schizophrenia in a healthy control group: psychometric properties and reference values. AB - BACKGROUND: Assessment of depression in schizophrenia is of great importance as depressive signs and symptoms and suicidality are highly prevalent in patients with schizophrenic disorders. The Calgary Depression Rating Scale (CDSS) is the standard assessment instrument for that purpose due to its proven reliability and validity. However, so far no reference values derived from an adequate healthy sample have been published. METHODS: The present study analyzed CDSS item scores and summary scores in 154 healthy subjects (49% female, mean age 32.8+/-11.7 years) selected from studies as controls for schizophrenic patients. RESULTS: The total CDSS score was 2.6+/-2.7 (range 0-12), about 1/3 of healthy subjects had scores=0. Gender differences were observed with respect to "early awakening" and "suicidal ideation" with higher scores in females. Total CDSS scores (r=-0.17, P=0.03) and single items were slightly negatively correlated with age. The psychometric properties of the CDSS scale were satisfying (corrected item-score correlations r(tc)=0.33-0.61, internal consistency Cronbach's alpha=0.76, one dimensional factor structure). CDSS total scores showed a skewed distribution fitting the upper part of a normal distribution. Based on the empirical distribution of CDSS values, reference values were listed for the total group of healthy subjects, and for females and males separately. CONCLUSIONS: The results corroborate the favourable psychometric properties of the CDSS found previously in clinical samples in a healthy control group. The reference values reported here can assist the clinical use and interpretation of CDSS values and emphasize the usefulness of the CDSS for a thorough assessment of depression and suicidality in schizophrenia. PMID- 16040127 TI - Components of self-esteem in affective patients and non-psychiatric controls. AB - Decrease in self-esteem (SE) is found in all mood disorders during inter-episode phases. This trait was associated with relapse and suicidality but its genetic basis is still undefined, probably because SE has multiple components. The aim of the current study was to ascertain which of those components were altered in a sample of affective patients. Three hundred and thirty-one outpatients with bipolar (N=199) and major depressive MD (N=132) disorders in remission for at least three months and one hundred controls completed the Rosenberg Self-esteem Scale (RSE; [Rosenberg, M., 1965. The measurement of self-esteem, Society and the Adolescent Self-Image. Princeton University Press, pp.16-36]). Principal component analysis was performed to identify RSE factor structure. Extracted factors were compared across case and control groups in the whole sample (N=431) and in a sub-sample (N=301) with low self-esteem (RSE <20). PCA yielded a two factor solution with self-confidence (SC) and self-deprecation (SD) that was largely consistent with the existing literature. Such factors were both associated with lower scores in affective patients than controls (SC: F=52, p<0.01; SD: F=43, p<0.01). However in the low RSE group only self-confidence was found to be decreased in subjects with mood disorders (SC: F=13.8, p<0.01; SD: F=0.05, p=0.9). These findings suggest that self-esteem deficit in affective disorders might involve specific components. Implications for research and clinical practice are discussed. PMID- 16040128 TI - Effects of candidate vaginally-applied microbicide compounds on innate immune cells. AB - Ideally, a vaginally-applied microbicide would be effective against a broad range of pathogens but would have minimal effects on the female genital tract. The aim of this study was to determine if representative candidate detergent-type and sulfated/sulfonated polymer-type microbicides altered the composition or function of innate immune cells normally found in the vaginal mucosa. The effect of microbicide on the composition of vaginal leukocytes was tested using a flow cytometric approach. Application of the detergent cholic acid, but not the sulfated polysaccharide lambda carrageenan, resulted in a significant increase in macrophages at the vaginal epithelial surface compared to control treatment (19.3% macrophages compared to 2.8%; p<0.0004). Phagocytosis of fluorochrome labeled bacteria by macrophages was inhibited greater than 50% in the presence of 1.0mg/ml of the sulfonated polymer PRO 2000 but was not inhibited by the same concentration of lambda carrageenan. PRO 2000-pulsed macrophages regained phagocytic function after being washed free of the compound. Culture of macrophages with PRO 2000 also resulted in diminished detection of the surface proteins CD11b and CD18. After treated cells were washed free of PRO 2000, these proteins were detected at levels similar to control treated cells. In conclusion, application of a detergent-type microbicide, but not a sulfated polymer, resulted in the infiltration of inflammatory cells at the vaginal epithelial surface. Phagocytic function of macrophages was lost in the presence of 1mg/ml PRO 2000 which may have reflected masking of important cell surface proteins by the microbicide; however, there was no evidence of permanent loss of function upon removal of the compound. PMID- 16040129 TI - Characterization of a monoclonal antibody identifying a CD45RA antigen on feline leukocytes. AB - The antibody produced by a murine hybridoma obtained from the fusion of SP2/0 plasmacytoma cells with splenocytes of a mouse immunized with feline bone marrow was found to react with 60% of bone marrow cells and 80% of peripheral blood leukocytes (PBL); reactivity in the latter tissue was restricted almost entirely to mononuclear cells. Two-color FACScan analyses of this antibody with mAbs specific for feline lymphocytes revealed positive and negative populations of CD4 and CD8 cells. The reactivity for CD4 and CD8 cells was animal age dependent, binding to a higher percentage of the cells in young (2-9 months) versus older animals (> 4 years). In a mitogen driven assay for IgG production by PBL the addition of this antibody to the cultures enhanced the suppressor activity of CD8 cells, a function attributed to activation of a CD4 suppressor-inducer population; removal of CD8 cells negated any induction of suppression. Mild papain digestion of bone marrow and PBL completely removed the antigen detected by this antibody while not affecting reactivity of a pan-T antibody. Western blot analysis showed binding of the antibody to polypeptides of approximately 200 kDa on feline bone marrow and PBL. The data suggest that this mAb is identifying the feline homologue of the leukocyte common antigen of cells with a functional specificity characteristic of a CD45RA isoform. PMID- 16040130 TI - Mast cells and IgE-bearing cells in lungs of RAO-affected horses. AB - Recurrent airway obstruction (RAO) is a common condition in stabled horses characterised by small airway inflammation and obstruction following exposure of susceptible horses to mouldy hay and straw. The aim of the present study was to investigate whether lung tissue from horses with RAO contains higher numbers of IgE-protein positive (+) cells and mast cells compared to controls after mouldy hay challenge. Furthermore, mast cell subtypes in lung tissue were investigated. IgE+ cells were detected in most lung tissue samples but no significant differences between RAO-affected and control horses were found. In the wall of the bronchi and bronchioli of both RAO-affected and control horses, mainly chymase+ mast cells (MC(C)) were present (85% in the bronchial wall and 77% in the wall of the bronchioli), while 73% of the mast cells (MC) around blood vessels were tryptase+ mast cells (MC(T)). No double stained MCs were detected. RAO-affected horses had significantly more MC(C) than controls in the wall of the bronchi (median=7.6 and 1.7 cell/mm(2), respectively, P< or =0.05). They also showed a tendency for more MC(C) in the wall of the bronchioli than controls (median=21 and 2.9 cells/mm(2), respectively, P=0.07) but there were no differences in MC(T) numbers. The data suggest an involvement of MC(C) in the pathogenesis of RAO. Independently of the clinical diagnosis, there was a significant relationship between high MC(C) numbers in the bronchial wall and lung fibrosis, suggesting that these MC(C) may be involved in tissue remodelling. Furthermore, high MC(C) numbers were also associated with increased infiltration with lymphocytes and neutrophils. PMID- 16040131 TI - Plasma levels of soluble adhesion molecules sPECAM-1, sP-selectin and sE-selectin are associated with relapsing-remitting disease course of multiple sclerosis. AB - Adhesion molecule mediated leukocyte migration into the central nervous system is considered to be a critical step in the pathogenesis of multiple sclerosis (MS). We measured plasma levels of the soluble adhesion molecules sPECAM-1, sP-selectin and sE-selectin in 166 MS patients and in 36 healthy blood donors with ELISA. sPECAM-1, sP-selectin and sE-selectin plasma concentrations showed a significant increase in the relapsing-remitting disease course of MS and were elevated during relapse. These findings indicate that sPECAM-1, sP-selectin and sE-selectin might be implemented as paraclinical markers of disease activity in MS with restriction to the clinical course of the disease. PMID- 16040132 TI - TNF-related apoptosis inducing ligand (TRAIL) gene polymorphism in Japanese patients with multiple sclerosis. AB - TNF-related apoptosis inducing ligand (TRAIL) has been reported to induce apoptosis of autoreactive T cells and other inflammatory cells, and thus, it is a strong candidate gene for involvement in the development of autoimmune diseases. We investigated single nucleotide polymorphisms (SNPs) in the coding region of the gene at position 1595 in exon 5 in 128 Japanese patients with conventional/classical multiple sclerosis (MS) and 158 healthy controls. Patients with optico-spinal MS (OSMS) or atypical clinical attacks were excluded from the study. The frequency of CC genotype at position 1595 was significantly different between patients and controls (p=0.0027), and the C allele was more prevalent in the patients than in the controls (p=0.0138, OR=1.546, 95% CI=1.092-2.188). Logistic analysis, adjusted for HLA-DRB1*1501-positivity, revealed the independent association of the CC genotype with susceptibility to MS (p=0.0006, OR=2.393, 95% CI=1.453-3.943). There were no significant associations between +1595 polymorphism and the clinical features of MS. The results indicate that the presence of the CC genotype at position 1595 in exon 5 represents a higher risk of MS. PMID- 16040133 TI - Altered populations of natural killer cell and natural killer T cell subclasses in myasthenia gravis. AB - Since the innate immune system can influence the disease activity of myasthenia gravis (MG), such as during infection, the frequencies of natural killer (NK) cells and NKT cells were analyzed in the blood and thymus. Before therapy (thymectomy plus glucocorticoid), the MG patients with thymic hyperplasia, but not those with thymoma, showed increased frequencies of mature NKT cells (CD3(+)TCRV(alpha)24(+)CD161(bright)) in the blood, while the frequency of immature NKT cells was unaltered. In the blood of the patients with thymoma, but not those with hyperplasia, the frequency of cytotoxic subclass of NK cells (CD3( )CD16(+)CD56(dim)) was lower than that of the control. These alterations returned to normal after therapy. The thymic frequencies of NKT cells and NK cells in MG thymuses were unaltered. These results suggest the involvement of both innate and acquired immunity in the disease activity of MG. PMID- 16040134 TI - Purification of capsid-like particles of infectious bursal disease virus (IBDV) VP2 expressed in E. coli with a metal-ion affinity membrane system. AB - Protein VP2, matured from the polyprotein encoded by the genome of infectious bursal disease virus (IBDV) and the primary host-protective immunogen of this virus, together with its two N-terminal truncated mutants were cloned and expressed in Escherichia coli. To obtain pure recombinant proteins for the development of an efficient subunit vaccine against IBDV infection, these three proteins were fused with six additional histidine residues at their C-termini as a His-purification-tag. Following purification employing immobilized metal-ion (Ni2+) affinity chromatography (IMAC), a purification fold of approximately 104 was achieved. Electron microscopic observation also demonstrated that all three E. coli-derived proteins form the morphology of icosahedral particles of approximately 25 nm in diameter. To reduce the cost of resin used for IMAC, self prepared immobilized metal-ion affinity membranes (IMAM), i.e., commercial, regenerated cellulose membrane modified with iminodiacetic acid and immobilized with nickel ions, was applied to purify particles formed by these three proteins. A 104-fold of purification efficiency was also achieved by this membrane, showing that under the same conditions the recovery and purification efficiency of IMAM are comparable with those of IMAC. The pure VP2-formed particles thus obtained, coupled with their uniform dimensions, not only facilitate a better understanding of the structural biology of these immunogenic particles but also help the development of improved vaccines against this avian virus. PMID- 16040135 TI - Surveillance technology for HIV-1 subtype C in Ethiopia: an env-based NASBA molecular beacon assay to discriminate between subcluster C and C'. AB - Forty-nine samples with known C2V3 sequences were used for the evaluation of an env-based molecular beacon assay to distinguish between the two genetic subclusters C and C' which characterize the HIV-1 epidemic in Ethiopia. Two subcluster C and two subcluster C' beacons targeting two different loci in the C2V3 region were developed. Using a three beacon-based (2C and 1C'=C prime), isothermal amplification assay, concordance with DNA sequencing was achieved for 43 (87.8%) samples. Sensitivity was 81.8% and specificity 97.4% for subcluster C beacons. For the subcluster C' beacon, a sensitivity of 97% and a specificity of 87.5% was achieved. Five samples were ambiguous by sequencing of which two samples were subcluster C' by the beacon assay and one subcluster C. Two of the samples remained ambiguous with different beacon-pair combinations as well. From samples with a clear C or C' phylogeny by sequencing, three were undetected by the first-line beacon genotyping assay. Genotype ambiguity was resolved in the three samples using beacon pair combinations restricted to each targeted locus. The beacons were evaluated further in a panel including all HIV-1 subtypes. Four of five subtype C isolates were identified correctly, and no cross-reactivity was observed with other subtypes. PMID- 16040136 TI - Activity-dependent modulation of the BDNF receptor TrkB: mechanisms and implications. AB - Although brain-derived neurotrophic factor (BDNF) has emerged as a key regulator of activity-dependent synaptic plasticity, a conceptually challenging question is how this diffusible molecule achieves local and synapse-specific modulation. One hypothesis is that neuronal activity enhances BDNF signaling by selectively modulating TrkB receptors at active neurons or synapses without affecting receptors on neighboring, less-active ones. Growing evidence suggests that neuronal activity facilitates cell-surface expression of TrkB. BDNF secreted from active synapses and neurons recruits TrkB from extrasynaptic sites into lipid rafts, microdomains of membrane that are enriched at synapses. Postsynaptic rises in cAMP concentrations facilitate translocation of TrkB into the postsynaptic density. Finally, neuronal activity promotes BDNF-induced TrkB endocytosis, a signaling event important for many long-term BDNF functions. These mechanisms could collectively underlie synapse-specific regulation by BDNF. PMID- 16040137 TI - Extracts and molecules from medicinal plants against herpes simplex viruses. AB - Herpes simplex viruses (HSV-1 and -2) are important pathogens for humans, especially in the case of highly susceptible adults. Moreover, HSV-2 has been reported to be a high risk factor for HIV infection. Therefore, the discovery of novel anti-HSV drugs deserves great efforts. In this paper, we review anti-HSV substances from natural sources, including both extracts and pure compounds from herbal medicines, reported in studies from several laboratories. The role of traditional medicine for the development of anti-HSV compounds is also discussed. Interestingly, it was found that traditional medicines, like Ayurvedic, traditional Chinese (TCM), Chakma medicines, are good and potential sources for promising anti-HSV drugs. A second objective of this review is to discuss several anti-HSV compounds with respect to their structure-activity relationship (SAR). A large number of small molecules, like phenolics, polyphenols, terpenes (e.g., mono-, di-, tri-), flavonoids, sugar-containing compounds, were found to be promising anti-herpetic agents. Our major conclusion is that natural products from medicinal plant extracts are very important source of anti-HSV agents. PMID- 16040138 TI - Improvement of heavy metal stress and toxicity assays by coupling a transgenic reporter in a mutant nematode strain. AB - Previous studies have demonstrated that wild type Caenorhabditis elegans displays high sensitivity to heavy metals in a lethality test at a level comparable to that of other bioindicator organisms. Taking advantage of the genetics of this model organism, we have tested a number of mutant strains for enhanced sensitivity in heavy metal induced lethality and stress response. These mutants are defective in genes controlling dauer formation, longevity or response to reactive oxygen species (ROS). Among the tested mutants, a double mutant daf-16 unc-75 strain was identified to have superior sensitivity. It has a 6-, 3- and 2 fold increase in sensitivity to cadmium, copper and zinc, respectively, as compared with that of wild type animals. When a fluorescent reporter transgene was coupled with this double mutant for stress detection, a 10-fold enhancement of sensitivity to cadmium over the wild type strain was observed. These transgenic animals, superior to most of the model organisms currently used in bioassays for environmental pollutants, offer a fast and economic approach to reveal the bioavailability of toxic substance in field samples. This study also demonstrates that combination of genetic mutations and transgenesis is a viable approach to identify sensitive indicator animals for environmental monitoring. PMID- 16040139 TI - Impact of cadmium contamination and oxygenation levels on biochemical responses in the Asiatic clam Corbicula fluminea. AB - The aim of this work was to evaluate the potential utility of several biochemical parameters as indicators of the toxic effects of cadmium in the freshwater clam Corbicula fluminea under two levels of oxygenation (normoxia 21 kPa and hypoxia 4 kPa). These variations in oxygenation are representative of the natural environments of bivalves living at the bottom of the water column, where hypoxic episodes may occur regularly. Cadmium accumulation, metallothionein synthesis, MXR protein induction, lipoperoxidation and antioxidant enzyme activities (catalase, glutathione reductase and total and selenium-dependent glutathione peroxidases) were assessed in the gills of C. fluminea in four experimental conditions: normoxia, hypoxia, normoxia with cadmium and hypoxia with cadmium ([Cd]=30 microg l(-1)) over a 14-day period. Behavioural reactions were also followed for the duration of the experiment by monitoring clam activity and valve movements. This study is a first report on biochemical responses under cadmium contamination and hypoxia and will enable us to determine better biomarkers for C. fluminea as they were measured simultaneously. In metal-exposed animals, we found an increasing accumulation of cadmium in the gills with time, and this was more severe in hypoxic conditions. Metallothionein synthesis occurred in contaminated clams and was precocious in hypoxic conditions. MXR protein induction appeared promising due to its quick and significant response to metal with a strong impact from hypoxic contamination. On the other hand, in our experimental conditions, antioxidant parameters did not show decisive responses to contamination and hypoxia, except glutathione peroxidases which decreased systematically with time in a cadmium-independent manner. Lipid peroxidation, expressed as malondialdehyde content, was not stimulated by normoxic contamination, as has been shown in other studies, but was stimulated under hypoxic cadmium contamination. Our study confirms the importance of a multi biomarker approach in environmental studies as some are not appropriate to all organisms. PMID- 16040140 TI - Effect of peripherally administered ghrelin on gastric emptying and acid secretion in the rat. AB - Ghrelin is a gut peptide that is secreted from the stomach and stimulates food intake. There are ghrelin receptors throughout the gut and intracerebroventricular ghrelin has been shown to increase gastric acid secretion. The aim of the present study was to examine the effects of peripherally administered ghrelin on gastric emptying of a non-nutrient and nutrient liquid, as well as, basal and pentagastrin-stimulated gastric acid secretion in awake rats. In addition, gastric contractility was studied in vitro. Rats equipped with a gastric fistula were subjected to an intravenous infusion of ghrelin (10-500 pmol kg(-1) min(-1)) during saline or pentagastrin (90 pmol kg( 1) min(-1)) infusion. After administration of polyethylene glycol (PEG) 4000 with 51Cr as radioactive marker, or a liquid nutrient with (51)Cr, gastric retention was measured after a 20-min infusion of ghrelin (500 pmol kg(-1) min(-1)). In vitro isometric contractions of segments of rat gastric fundus were studied (10( 9) to 10(-6) M). Ghrelin had no effect on basal acid secretion, but at 500 pmol kg(-1) min(-1) ghrelin significantly decreased pentagastrin-stimulated acid secretion. Ghrelin had no effect on gastric emptying of the nutrient liquid, but significantly increased gastric emptying of the non-nutrient liquid. Ghrelin contracted fundus muscle strips dose-dependently (pD2 of 6.93+/-0.7). Ghrelin IV decreased plasma orexin A concentrations and increased plasma somatostatin concentrations. Plasma gastrin concentrations were unchanged during ghrelin infusion. Thus, ghrelin seems to not only effect food intake but also gastric motor and secretory function indicating a multifunctional role for ghrelin in energy homeostasis. PMID- 16040141 TI - Thyrotropin-releasing hormone in the dorsal vagal complex stimulates pancreatic blood flow in rats. AB - Central administration of thyrotropin-releasing hormone (TRH) enhanced pancreatic blood flow in animal models. TRH nerve fibers and receptors are localized in the dorsal vagal complex (DVC), and retrograde tracing techniques have shown that pancreatic vagal nerves arise from the DVC. However, nothing is known about the central sites of action for TRH to elicit the stimulation of pancreatic blood flow. Effect of microinjection of a TRH analog into the DVC on pancreatic blood flow was investigated in urethane-anesthetized rats. After measuring basal flow, a stable TRH analog (RX-77368) was microinjected into the DVC and pancreatic blood flow response was observed for 120 min by laser Doppler flowmetry. Vagotomy of the several portions, or pretreatment with atoropine methyl nitrate or N(G) nitro-l-arginine-methyl ester was performed. Microinjection of RX-77368 (0.1-10 ng) into the left or right DVC dose-dependently increased pancreatic blood flow. The stimulation of pancreatic blood flow by RX-77368 microinjection was eliminated by the same side of cervical vagotomy as the microinjection site or subdiaphragmatic vagotomy, but not by the other side of cervical vagotomy. The TRH-induced stimulation of pancreatic blood flow was abolished by atropine or N(G)-nitro-l-arginine-methyl ester. These results suggest that TRH acts in the DVC to stimulate pancreatic blood flow through vagal-cholinergic and nitric oxide dependent pathways, indicating that neuropeptides may act in the specific brain nuclei to regulate pancreatic function. PMID- 16040142 TI - The effects of short term (3 weeks) testosterone treatment on serum inflammatory markers in men undergoing coronary artery stenting. AB - OBJECTIVE: Inflammation markers can predict restenosis after successful intracoronary stenting. There is evidence that testosterone suppresses the expression of the inflammatory cytokines. We hypothesized that testosterone therapy after coronary stenting can reduce the inflammation markers. METHODS: We selected 41 men with coronary artery disease who underwent successful stent implantation for a >70% diameter stenosis of a major coronary artery. Patients, who had stable angina and positive exercise test results, were recruited after diagnostic coronary angiography. Twenty-five men were treated with 3 doses of i.m. testosterone administration once a week for 3 weeks following diagnostic angiography. Sixteen patients were recruited as a control group and they received standard therapy. First venous blood samples were obtained after angiography. Stents were implanted 3 weeks after diagnostic angiography. Second venous blood samples were taken 24 h after the coronary stenting. RESULTS: Baseline biochemical or hematological parameters were similar between the control and treatment groups. After coronary stenting, free testosterone, total testosterone, and sex hormone binding globulin were significantly elevated in the testosterone group (P<0.0001, P<0.0001 and P=0.02; respectively). After coronary stent implantation, there was a significant increase in IL-6 and CRP levels in the control group only (P=0.02 and P=0.038), while TNF-alpha levels were increased significantly in both groups (P=0.016 and P=0.014; respectively). Statistical analysis revealed that testosterone treatment prior to stent implantation attenuated IL-6 and hs-CRP levels significantly (P=0.042 and P=0.043; respectively). CONCLUSIONS: The present study shows that 3 weeks testosterone treatment prior to intracoronary stenting results in a significant suppression in hs-CRP and IL-6 levels after the stent implantation. PMID- 16040143 TI - Quantification of metabolically active biomass using Methylene Blue dye Reduction Test (MBRT): measurement of CFU in about 200 s. AB - Quantification of viable cells is a critical step in almost all biological experiments. Despite its importance, the methods developed so far to differentiate between viable and non-viable cells suffer from major limitations such as being time intensive, inaccurate and expensive. Here, we present a method to quantify viable cells based on reduction of methylene blue dye in cell cultures. Although the methylene blue reduction method is well known to check the bacterial load in milk, its application in the quantification of viable cells has not been reported. We have developed and standardized this method by monitoring the dye reduction rate at each time point for growth of Escherichia coli. The standard growth curve was monitored using this technique. The Methylene Blue dye Reduction Test (MBRT) correlates very well with Colony Forming Units (CFU) up to a 800 live cells as established by plating. The test developed is simple, accurate and fast (200 s) as compared to available techniques. We demonstrate the utility of the developed assay to monitor CFU rapidly and accurately for E. coli, Bacillus subtilis and a mixed culture of E. coli and B. subtilis. This assay, thus, has a wide applicability to all types of aerobic organisms. PMID- 16040144 TI - Probing drug action using in vitro pharmacological profiles. AB - The history of structure-activity relationships in drug design represents a long search for appropriate descriptors of broad biological action at the molecular level. In this context, recent work showing that in vitro pharmacological profiles can be used as exquisite descriptors of the broad biological effects of compounds represents an important breakthrough. Generalization of the methodology could have important implications for drug discovery and development. It might also provide a novel and insightful way to study systems biology. PMID- 16040146 TI - Post-awakening cortisol secretion during basic military training. AB - Salivary free cortisol concentrations in the first 30 min after awakening were assessed in 12 healthy army recruits at the beginning, middle and end of an 11 week intensive physical training course. To ensure strict adherence to protocol saliva collection was supervised and collected on each sampling day immediately on waking and again 15 and 30 min later. Self-rated psychological assessments of state levels of stress, arousal and fatigue were performed in the evening of each sampling day. A within-subjects repeated-measures analysis of participants who completed the course (12 of the original 20) showed a significant main effect of cortisol concentration across all three sampling points after awakening (F((2,22))=54.516, p<0.0001) and a significant main effect of weeks into the training course (F((3,33))=4.390, p=0.010). Further analysis of this effect of measurement-week revealed that at weeks 3 and 6 total cortisol secretion estimated by area under the curve was lower (F((3,33))=4.602, p=0.008) compared to the beginning and end of the course. Surprisingly self-reported stress, arousal and fatigue did not differ significantly across weeks, despite the large dropout rate (40%) and self-evident pressures of the course. We conclude that when controlling for many confounding variables, including participant adherence, post-awakening cortisol levels are sensitive to stressful challenge over a period of weeks. PMID- 16040147 TI - Functional and morphological organization of the nucleus tractus solitarius in the fictive cough reflex of guinea pigs. AB - Projection of the superior laryngeal nerve (SLN) afferent fibers into the nucleus tractus solitarius (NTS) was investigated using a fluorescent tracer in guinea pigs. High density of fluorescence was detected in the ipsilateral NTS extending from 0.5 mm caudal to 1.2 mm rostral to the obex. At coronal slices, the fluorescent granules, lines and patches were located in the interstitial, medial and dorsal regions of NTS. Fluorescence was also found in the dorsal region of contralateral commissural NTS. Microstimulation of the rostral NTS, which corresponded to the region showing the strong fluorescence, induced an increase in the inspiratory discharge of phrenic nerve that was immediately followed by a large burst discharge of the iliohypogastric nerve in decerebrate, paralyzed and artificially ventilated guinea pigs. This serial response of the two nerves was identical to that induced by electrical stimulation of the SLN. Intravenous injection of codeine suppressed both NTS and SLN-induced responses. The SLN induced response was inhibited by microinjection of codeine into the ipsilateral NTS and abolished by lesion of the ipsilateral NTS. These results suggest that the NTS has an integrative function in production of cough reflex and is possible sites of action of central antitussive agents. PMID- 16040148 TI - Application of in vitro methods for selection of Lactobacillus casei strains as potential probiotics. AB - Probiotics have established their efficacy as dietary adjuncts providing benefits to consumers, but the selection of probiotics before incorporation in diet requires close scrutiny in the form of in vitro as well as in vivo tests. The present study was undertaken to check different in vitro characteristics of seven Lactobacillus casei strains. The characteristics studied include acid and bile tolerance, adhesion and cell surface hydrophobicity, antimicrobial effect on common pathogens and cholesterol reduction. All strains were able to resist pH 3 for 3 h, though resistance to pH 2 was exhibited by NCDC 17, C1 and Y strains only. NCDC 63 and VT strains were able to tolerate 1% and 2% bile concentrations for 12 h. There was wide variation in ability of strains to adhere to isolated rat epithelial cells. The index was highest for C1 at 66%. The electron microscopic adhesion studies on the stainless steel chips did not reveal any specific attachment to surfaces by any of strains. The hydrophobic character for octane was highest for strain C1 at 54.06% and lowest for strain C2 at 4.65%. The ability to antagonize common pathogens was observed in all strains but this activity was attributed to production of organic acids and no specific compound caused the inhibitory effect. The cholesterol reducing ability varied not only for strains but also for time of incubation. NCDC 17 showed maximum reduction in cholesterol level after 48 h of incubation with buffalo plasma as the source of cholesterol. Overall there existed variations in different strains with respect to different characters of significance to be a probiotic. PMID- 16040149 TI - Mycobacterial manipulation of the host cell. AB - Phagosome biogenesis, the process by which macrophages neutralize ingested pathogens and initiate antigen presentation, has entered the field of cellular mycobacteriology research largely owing to the discovery 30 years ago that phagosomes harboring mycobacteria are refractory to fusion with lysosomes. In the past decade, the use of molecular genetics and biology in different model systems to study phagosome biogenesis have made significant advances in understanding subtle mechanisms by which mycobacteria inhibit the maturation of its phagosome. Thus, we are beginning to appreciate the extent to which these pathogens are able to interfere with innate immune responses and manipulate defense mechanisms to enhance their survival within the human host cell. Here, we summarize current knowledge about phagosome maturation arrest in infected macrophages and the subsequent attenuation of the macrophage-initiated adaptive anti-mycobacterial immune defenses. PMID- 16040150 TI - Purine and pyrimidine transport in pathogenic protozoa: from biology to therapy. AB - Purine salvage is an essential function for all obligate parasitic protozoa studied to date and most are also capable of efficient uptake of preformed pyrimidines. Much progress has been made in the identification and characterisation of protozoan purine and pyrimidine transporters. While the genes encoding protozoan or metazoan pyrimidine transporters have yet to be identified, numerous purine transporters have now been cloned. All protozoan purine transporter-encoding genes characterised to date have been of the Equilibrative Nucleoside Transporter family conserved in a great variety of eukaryote organisms. However, these protozoan transporters have been shown to be sufficiently different from mammalian transporters to mediate selective uptake of therapeutic agents. Recent studies are increasingly addressing the structure and substrate recognition mechanisms of these vital transport proteins. PMID- 16040151 TI - Interleukin-6 plays a crucial role in the hepatic expression of SOCS3 during acute inflammatory processes in vivo. AB - BACKGROUND/AIMS: Interleukin-6 is mandatory for liver regeneration after injury and for the hepatic expression of acute phase proteins and cytochrome P450 enzymes during inflammation. Due to its crucial contribution to the maintenance of homeostasis IL-6 signaling is tightly controlled. Suppressor of cytokine signaling (SOCS) 3 is a potent IL-6-induced feedback inhibitor terminating IL-6 signal transduction. However, several signaling pathways converge on SOCS3: SOCS3 can be induced by other mediators in vitro, and it does not exclusively inhibit IL-6 signaling. The individual contribution of each cytokine to the induction of SOCS3 in vivo is unknown. METHODS: Using IL-6-deficient mice we analyzed the role of interleukin-6 for the hepatic SOCS3 expression in response to turpentine and LPS as models of aseptic and bacterial inflammation, respectively. RESULTS: In wild-type animals, turpentine and LPS elicited strong induction of SOCS3. IL-6 deficient mice, by contrast, showed severely impaired SOCS3 expression in response to both stimuli: turpentine failed to induce SOCS3 mRNA; in LPS-induced inflammation, the early inductive response 60min after LPS injection was absent, and the delayed expression of SOCS3 was markedly reduced. The residual delayed SOCS3 expression in IL-6-deficient mice was abolished in IL-6/TNFR-1 knockout mice. CONCLUSIONS: Our data strongly argue for a crucial role of IL-6 in the hepatic expression of SOCS3 during acute inflammatory processes in vivo. Although other cytokines are capable of inducing SOCS3 their contribution seems to be minor. PMID- 16040152 TI - Delayed transfers of older people from hospital: Causes and policy implications. AB - Health and social care agencies in the UK. have been under pressure for some time to reduce delayed transfers of older people from hospital because they absorb scarce health service resources and incur a human cost through inappropriate placement. A local study based on an analysis of records and interviews with managers showed that delays reflect the complex needs of older people, and arise from financing and organisational problems at both the planning and implementation stages of a discharge. Family resistance may also be a factor. Budgetary constraints result in delays in confirming public support for some clients. Shortages of professional staff and care assistants limit the provision of domiciliary packages. The contraction of the residential sector has reduced the availability of beds and increased the cost of care home placements. Scope exists for expediting administrative aspects of transfers by coordinating health and social services. More recent legislation that imposes fines on social service departments for delayed transfers does not address underlying causes. PMID- 16040153 TI - Computerized segmentation of whole-body bone scintigrams and its use in automated diagnostics. AB - Bone scintigraphy or whole-body bone scan is one of the most common diagnostic procedures in nuclear medicine used in the last 25 years. Pathological conditions, technically poor image resolution and artefacts necessitate that algorithms use sufficient background knowledge of anatomy and spatial relations of bones in order to work satisfactorily. A robust knowledge based methodology for detecting reference points of the main skeletal regions that is simultaneously applied on anterior and posterior whole-body bone scintigrams is presented. Expert knowledge is represented as a set of parameterized rules which are used to support standard image-processing algorithms. Our study includes 467 consecutive, non-selected scintigrams, which is, to our knowledge the largest number of images ever used in such studies. Automatic analysis of whole-body bone scans using our segmentation algorithm gives more accurate and reliable results than previous studies. Obtained reference points are used for automatic segmentation of the skeleton, which is applied to automatic (machine learning) or manual (expert physicians) diagnostics. Preliminary experiments show that an expert system based on machine learning closely mimics the results of expert physicians. PMID- 16040154 TI - Sampling schedule design towards optimal drug monitoring for individualizing therapy. AB - We study the individualization of therapy by simultaneously taking into account the design of sampling schedule and optimal therapeutic drug monitoring. The sampling schedule design in this work is to determine the number of samples, the sampling times, the switching time from the loading to the maintenance period, and the drug dosages. A closed-loop control policy is employed to determine the sampling schedule, and an advanced stochastic global optimization algorithm, which integrates the stochastic approximation and simulated annealing techniques, is implemented to search the optimal sampling schedule. A simulated one compartment model of intravenous theophylline therapy is used to illustrate our method. This method can be readily extended to multiple compartment systems and allow incorporating other criteria of drug control. While currently the method is mainly of theoretical interest, it offers a starting point for practical applications and thus is hopefully of great value for the clinically individualizing therapy in the future. PMID- 16040155 TI - Heterogeneity of chlorinated hydrocarbon sorption properties in a sandy aquifer. AB - Hydraulic conductivity and sorption coefficients for chlorinated hydrocarbons (chloroform, carbon tetrachloride and tetrachloroethylene) were evaluated for 216 sediment samples collected across a 15 m transect and a 21 m depth interval in a contaminated aquifer near Schoolcraft, Michigan. Relationships between hydraulic conductivity, linear sorption partition coefficients, grain size classes, and spatial location were investigated using linear regression analysis and geostatistical techniques. Clear evidence of layering was found in sorption properties, hydraulic conductivity and grain sizes. Conductivity correlated well with grain size, as expected, but sorption varied inversely with grain size, contrary to some previous reports. No significant correlation was found between sorption properties and hydraulic conductivity. This is likely due to the unexpected presence of small amounts of highly sorptive coal-like solids, which dominate the sorption behavior but have little effect on conductivity. The results demonstrate that recent findings regarding the high sorption capacity of coal materials found in soils can exert a controlling influence on contaminant transport. Designers of in situ remediation systems should be cautioned that 1) it is not reasonable to assume that sorption capacity and hydraulic conductivity are related, 2) sorption capacity and hydraulic conductivity are critical measurements for contaminant site characterization and subsequent transport modeling, 3) estimating sorption capacity from organic carbon measurement may lead to greater errors than performing sorption isotherms, and 4) it is more important to characterize vertical heterogeneity rather than horizontal heterogeneity because both sorption and hydraulic conductivity are correlated across longer distances in the horizontal plane. PMID- 16040156 TI - Effects of neuropeptide Y antagonists on food intake in rats: differences with cold-adaptation. AB - Hyperphagia followed both central neuropeptide Y (NPY) administration and the presumed increase of endogenous NPY activity after food deprivation. NPY induced greater hyperphagia in cold-adapted than non-adapted rats; fasting of comparable severity caused similar hyperphagia in the two groups. NPY-receptor-antagonist D Tyr(27,36), D-Thr32-NPY(27,36) or functional NPY-antagonist D-myo-inositol-1,2,6 trisphosphate attenuated the hyperphagic effect of both NPY and fasting in non adapted rats. However, while completely preventing the NPY-hyperphagia, they did not influence the fasting-induced hyperphagia in cold-adapted rats. With cold adaptation the sensitivity to NPY and to its antagonists increases, but the hypothalamic NPY loses from its fundamental role in the regulation of food intake, and the hyperphagia seen in cold-adaptation may need some other explanation. PMID- 16040157 TI - Genomic organization of four novel nondisulfide-bridged peptides from scorpion Mesobuthus martensii Karsch: gaining insight into evolutionary mechanism. AB - At least 25 nondisulfide-bridged peptides (NDBPs) have been identified and characterized from scorpions. However, the genomic organization of the genes that encode these peptides have not been reported yet. BmKa1, BmKa2 and BmKb1 are three novel genes that code for NDBPs identified by our group from Mesobuthus martensii Karsch. Based on their cDNA sequences, the genomic DNA sequences encoding these peptides were obtained using the PCR method. Sequence analysis showed that three distinct genomic structural patterns are used to encode these three peptides. The BmKa1 gene is not interrupted by any introns. However, the BmKa2 gene is composed of two exons, interrupted by a 67 bp intron that is located in the DNA region encoding the mature peptide. Two genomic homologues of the BmKb1 cDNA sequence, named BmKb1' and BmKb2, respectively, were obtained. The BmKb1' gene contains one intron of 593 bp, inserted into the DNA region that encodes the signal peptide. Similarly, the BmKb2 gene also contains an intron that interrupts the exon that encodes the NDBP signal peptide. The amino acid sequences deduced for BmKb2 and BmKb1' differ only at one position. The data suggest that the genomic organizational pattern of NDBPs displays more divergence than that exhibited by the genes that encode disulfide-bridged peptides from scorpions. PMID- 16040158 TI - Interaction between adenosine A 2B-receptors and alpha2-adrenoceptors on the modulation of noradrenaline release in the rat vas deferens: possible involvement of a group 2 adenylyl cyclase isoform. AB - In the prostatic portion of rat vas deferens, activation of adenosine A 2B receptors, beta2-adrenoceptors, adenylyl cyclase or protein kinase A caused a facilitation of noradrenaline release. Blockade of alpha2-adrenoceptors with yohimbine (1 microM) attenuated the facilitation mediated by adenosine A 2B receptors and by direct activation of adenylyl cyclase with forskolin but not that mediated by beta2-adrenoceptors or by direct activation of protein kinase A with 8-bromoadenosine-3',5'-cyclicAMP. The adenosine A 2B- and the beta2 adrenoceptor-mediated facilitation was prevented by the adenylyl cyclase inhibitors, 2',5'-dideoxy-adenosine (3 microM) and 9-cyclopentyladenine (100 microM), at concentrations that also attenuated the release enhancing effect of forskolin, but were not changed by the phospholipase C inhibitor 1-[6-[((17beta) 3-methoxyestra-1,3,5[10]-trien-17-yl)amino]hexyl]-1H-pyrrole-2,5-dione (U-73122, 1 microM). Facilitation of noradrenaline release mediated by adenosine A 2B receptors was also attenuated by activation of protein kinase C with the phorbol ester 12-myristate 13-acetate (1 microM) and by inhibition of Gbetagamma subunits with an anti-betagamma peptide; facilitation mediated by beta2-adrenoceptors was mainly attenuated by the calmodulin inhibitor calmidazolium (10 microM) and by the calmodulin kinase II inhibitor (N-[2-[N-(4-chlorocinnamyl)-N methylaminomethyl]phenyl]-N-(2-hydroxyethyl)-4-methoxybenzene-sulfonamide phosphate (KN-93, 5 microM). The results suggest that adenosine A 2B- but not beta2-adrenoceptor-mediated facilitation of noradrenaline release is enhanced by an ongoing activation of alpha2-adrenoceptors. They further suggest that adenosine A 2B-receptors and beta2-adrenoceptors are coupled to distinct adenylyl cyclase isoforms what may explain the different influence of alpha2-adrenoceptor signalling pathway on the facilitatory effects mediated by the two adenylyl cyclase coupled receptors. PMID- 16040159 TI - Synthesis and biological activity of novel acridinylidene and benzylidene thiazolidinediones. AB - A novel set of acridinylidene thiazolidinediones and benzylidene thiazolidinediones was synthesized by nucleophilic addition of cyanoacrylates. Some of these compounds were evaluated for their glucose lowering capability and their effects on the triglyceride level in alloxan diabetic mice. PMID- 16040160 TI - Facile synthesis of active antitubercular, cytotoxic and antibacterial agents: a Michael addition approach. AB - Spiro derivatives of oxindole and isoxazole-5-one were synthesized by using Michael addition reaction, highlighting the regioselective approach towards the synthesis of Michael diadduct followed by condensation of Michael diadduct. The spiro compound 4 showed antitubercular activity against Mycobacterium tuberculosis H37Rv whereas spiro compound 9 possesses pronounced anticancer and antibacterial profile. PMID- 16040161 TI - Novel milrinone analogs of pyridine-3-carbonitrile derivatives as promising cardiotonic agents. AB - In an attempt to design new inotropic drugs for congestive heart failure (CHF) with less proarrhythmic potential, three series of compounds analogous to milrinone were prepared, namely, 4-aryl-6-(4-pyridyl)-2-oxo-1,2-dihydropyridine-3 carbonitriles 2a-g, 4-aryl-6-(4-pyridyl)-2-thioxo-1,2-dihydropyridine-3 carbonitriles 3a-g and 2-amino-4-aryl-6-(4-pyridyl)-pyridine-3-carbonitriles 4a g. The first series was prepared by reacting 4-acetyl pyridine with the appropriate aldehyde, ethyl cyanoacetate and ammonium acetate in ethanol. Reaction of 2a-g with phosphorus pentasulfide afforded the second series 3a-g. The third target compounds 4a-g were prepared applying the same procedure used to synthesize 2a-g using malononitrile instead of ethyl cyanoacetate. All the newly synthesized compounds were evaluated for their cardiotonic activity and their in vivo cardiovascular effects. In addition, their oral and parenteral acute toxicity were determined. Compounds 2a, 2b, 2c, 4c and 4f proved to exert cardiotonic activity comparable to that of milrinone using spontaneously beating atria model from reserpine-treated guinea pigs. In addition these compounds proved to be non-toxic and well tolerated by mice up to 250 mg kg(-1) orally and up to 125 mg kg(-1) through parenteral route. PMID- 16040162 TI - Synthesis of some novel benzoxazole derivatives as anticancer, anti-HIV-1 and antimicrobial agents. AB - In an effort to establish new candidates with improved antineoplastic, anti-HIV-1 and antimicrobial activities we report here the synthesis and in vitro biological evaluation of various series of 2-substituted benzoxazoles: 2-[(Arylhydrazono, arylidene, cycloalkylidene and N-substituted thiocarbamoyl)cyanomethyl] benzoxazoles(2-4 and 7, respectively); 2-[(4- or 5-oxothiazoliden-2 yliden)benzoxazoles (5 and 6) and 2-(4-amino-3-substituted-2-thioxo-2,3 dihydrothiazol-5-yl)benzoxazoles (8), together with the synthesis of some substituted 3H-pyrido[2,1-b]benzoxazoles (9-11). The absolute configuration of compound 3b was determined by X-ray crystallography. The results of the in vitro anticancer screening revealed that some of the tested compounds exhibited broad spectrum antitumor activity. The most active compounds are 2a, 3b, 8a and 8d, their GI50 MG-MID values: 37.7, 19.1, 20.0 and 15.8 microM; TGI MG-MID values: 75.9, 53.7, 53.7, and 58.9 microM; and LC50 MG-MID values: 97.7, 93.3, 89.1 and 93.3 microM, respectively. The in vitro microbiological data showed that compound 7c was the most active against Staphylococcus aureus (minimal inhibitory concentration (MIC)<12.5 microg ml(-1). While compounds 5, 8a, and 8d were the most active against Bacillus subtilis (MIC values<12.5 microg ml(-1)). On the other hand, compounds 5 and 7c were the most active against Escherichia coli (MIC<25 microg ml(-1)), their activity is about half the activity of ampicillin and streptomycin . In addition, compound 4b and 7c were the most active against Pseudomonas aeruginosa (MIC<25, 50 microg ml(-1)). Compound 4b was two times as active as ampicillin and streptomycin while compound 7c was active as both. The results of antimycotic activity indicated that, Compound 7c showed mild activity against Candida albicans when compared with clotrimazole (MIC<100 microg ml(-1)). In vitro HIV-1 testing revealed that compound 7a displayed moderate anti-HIV-1 activity (maximum % cell protection, 36.6 at 2 x 10(-5) microM). PMID- 16040163 TI - Synthesis and structure-activity relationships of mono- and dialkyl-substituted oxaliplatin derivatives. AB - In order to improve the pharmacological profile of the anticancer drug oxaliplatin, (trans-R,R-cyclohexane-1,2-diamine)oxalatoplatinum(II), and to explore activity-structure relationships, new mono- and dialkyl substituted oxaliplatin analogues have been synthesized. Following a new synthetic strategy, racemates with a defined stereochemistry at carbon atoms 1, 2, 4, and 5 of the cyclohexane ring could be prepared, which are the bases for reliable structure activity relationships and the following enantiomer resolution. The cytotoxicity was evaluated in nine tumor cell lines, indicating that bulky substituents have a negative influence on the cytotoxic potency of the oxaliplatin derivatives. With respect to the antiproliferative properties, the 4-methyl-, cis-4,5-dimethyl-, and especially the 4,4-dimethyl-trans-cyclohexane-1,2 diamine(oxalato)platinum(II) complexes are the most promising candidates to be further evaluated. PMID- 16040164 TI - [Atherosclerosis and rheumatoid arthritis]. AB - AIMS: To identify studies which have shown that rheumatoid arthritis (RA) is associated with an increase in cardiovascular morbidity and mortality. To identify the different factors that may be involved. To consider what management would decrease the cardiovascular morbidity and mortality of RA. RESULTS: Epidemiological studies have shown that the risk of a cardiovascular event is increased twofold in RA patients irrespective of the traditional cardiovascular risk factors. Non-invasive methods have shown that RA patients have endothelial dysfunction, decreased arterial compliance and increased intima-media thickers, predictive factors for cardiovascular events in comparison to controls after controlling for traditional cardiovascular risk factors. The increased cardiovascular risk is directly mediated by inflammatory syndrome, which also indirectly increases the risk by inducing dyslipidemia and insulin resistance. Treatments also have a hamful effect, whether it be corticosteroid therapy, non steroidal anti-inflammatory drugs (NSAIDs), or methotrexate (MTX), which leads to hyperhromocysteinemia. CONCLUSION: It should be possible to decrease cardiovascular morbidity and mortality by a strict control of the disease's activity. We should also take measures to combat other cardiovascular risk factors: as low a dose as possible for corticosteroid therapy, limited prescription of NSAIDs, systematic supplementation of MTX with folic acid encouragement of smoking cessation, regular lipid tests and prescription of statins treatment for hyperlipemia in accordance with current recommendations. PMID- 16040165 TI - [Strokes associated with cervical artery dissection, and systemic mastocytosis: an unfortuitous association? A report of two cases]. AB - INTRODUCTION: Neuropsychiatric symptoms in systemic mastocytosis are usually cognitive and affective changes. EXEGESIS: We describe here two systemic mastocytosis patients without eosinophilia presenting strokes associated with cervical artery dissection. CONCLUSION: These observations are the first reported and they suggest that systemic mastocytosis could be add to the predisposing factors of spontaneous cervical artery dissections. PMID- 16040166 TI - Introducing problem-based learning into research methods teaching: student and facilitator evaluation. AB - The evidence base for the effectiveness of problem-based learning (PBL) has never been substantively established, although PBL is a generally accepted approach to learning in health care curricula. PBL is believed to encourage transferable skills, including problem-solving and team-working. PBL was used to deliver a postgraduate research methods module and a small evaluation study to explore its efficacy was conducted amongst the students (n = 51) and facilitators (n = 6). The study comprised of an evaluation questionnaire, distributed after each themed group of PBL sessions, and a group discussion conducted 4 weeks after the conclusion of the module, which was attended by student representatives and the facilitators. Questionnaire data was analysed using SPSS, and a transcript of the interview was subjected to content analysis. The results indicated that students felt that a PBL approach helped to make the subject matter more interesting to them and they believed that they would retain knowledge for a longer period than if their learning had used a more traditional lecture format. Students also perceived that PBL was effective in its ability to enhance students' understanding of the group process. All those involved in the PBL process reinforced the pivotal role of the facilitator. This study indicates that there is potential for PBL to be used beyond the more usual clinical scenarios constructed for health care professional education and further exploration of its use in areas such as building research capability should be undertaken. PMID- 16040167 TI - Emerging identity of Finnish nurse teachers: student teachers' narratives in a group exam. AB - In Finland students pursuing in Master's Degree Programme have a previous degree in nursing from polytechnics. Nurse teacher qualification also includes at least five year's experience of working in practice. This article concentrates on the conflicts student teachers brought up in a group exam as regards changing a nurse's identity into a teacher's identity in teacher training. The approach of the study on this group was narrative. The data were collected by videotaping two group exam situations (five hours in all) of volunteer student teachers (N= 5) during the last course (Nurse teacher's inquiring mind) of their nurse teacher studies in 2002. The material was transcribed and analysed using categorical content reading to identify particular themes, story lines and messages within material. By abstracting these, stories that describe the content can be found. Three narratives emerged in the analysis: 1. Competence creates confidence; 2. Shared teacher identity; 3. Ethics of teacher identity. The narratives raised new ideas for developing nurse teacher education in the future. Nowadays, there are good prospects for developing a shared teacher identity if there is willingness to co-operate within a learning community of a new kind. PMID- 16040168 TI - All-trans retinoic acid biases immune response induced by DNA vaccine in a Th2 direction. AB - Vitamin A deficiency diminishes Th2-mediated Ab responses. Providing Vitamin A or its active metabolites reverses this defect. All-trans retinoic acid (ATRA), an acid derivation of Vitamin A, regulates the balance of immune response induced by TR421-hCGbeta DNA vaccine. Compared to DNA vaccine alone or treatment with vehicle, significantly higher level of antibody against the protein encoded by DNA vaccine was observed in mice 6 weeks after the first immunization. The IgG2a/IgG1 ratio was lower in mice treated with ATRA. We also found that treatment with ATRA also diminishes specific cellular immune response induced by gene immunization by measuring the marker of cellular immune response. We conclude that ATRA biases the immune response to Th2 direction induced by DNA vaccine and acts as a candidate adjuvant and immunomodulatory molecule. PMID- 16040169 TI - Oral immunization with a shiga toxin B subunit: rotavirus NSP4(90) fusion protein protects mice against gastroenteritis. AB - A fusion protein containing the shiga toxin-1 B subunit (STB) linked to a 90 amino acid peptide (aa residues 86--175) from simian rotavirus (SA--11) nonstructural protein NSP4 was synthesized in Escherichia coli. Mice orally inoculated with 60 microg of STB::NSP4(90) fusion protein per dose generated higher humoral and intestinal antibody titers than mice inoculated with 30 microg of NSP4 alone. Serum anti-NSP4 IgG2a isotype titers were substantially greater than IgG1 titers, suggesting a dominant Th1 immune response. ELISA measurement of cytokines secreted from splenocytes isolated from immunized mice confirmed the STB::NSP4(90) fusion protein stimulation of a strong Th1 cell mediated immune response. Diarrhea in SA-11 rotavirus challenged neonates suckling from STB::NSP4 immunized dams was significantly reduced in severity and duration in comparison with virus challenged neonates from unimmunized mice. Together, our experiments demonstrate for the first time that the shiga toxin B subunit provides ligand mediated delivery of virus antigens to the gut-associated lymphoid tissues for enhanced stimulation of humoral and cellular responses against rotavirus gastroenteritis. PMID- 16040171 TI - Modelling the extra and intracellular uptake and discharge of heavy metals in Fontinalis antipyretica transplanted along a heavy metal and pH contamination gradient. AB - Samples of the aquatic bryophyte Fontinalis antipyretica Hedw. were transplanted to different sites with the aim of characterizing the kinetics of the uptake and discharge of heavy metals in the extra and intracellular compartments. The accumulation of metals in extracellular compartments, characterized by an initial rapid accumulation, then a gradual slowing down over time, fitted perfectly to a Michaelis-Menten model. The discharge of metals from the same compartment followed an inverse linear model or an inverse Michaelis-Menten model, depending on the metal. In intracellular sites both uptake and discharge occurred more slowly and progressively, following a linear model. We also observed that the acidity of the environment greatly affected metal accumulation in extracellular sites, even when the metals were present at relatively high concentrations, whereas the uptake of metals within cells was much less affected by pH. PMID- 16040172 TI - The landsnail Cepaea nemoralis regulates internal Cd levels when fed on Cd enriched stinging nettle (Urtica dioica) leaves at low, field-relevant concentrations. AB - We studied Cd accumulation in Cepaea nemoralis snails at low, but field-relevant Cd concentrations in the diet (Urtica dioica leaves). Six treatments of U. dioica plants were grown, resulting in leaf Cd concentrations between 0 and 2.6 microg g(-1) dw. Seven snails per treatment were fed for 38 days. Leaf Cd concentrations did not affect food consumption rates, and consequently Cd intake rates increased with increasing leaf concentrations. No differences were detected among treatments in the final soft tissue Cd concentrations and body burdens in the snails. Regression analyses showed no positive relationship between either snail Cd concentrations or body burdens and total Cd intake. This suggests a regulation of internal Cd concentrations at low food Cd concentrations. Our data suggest that Cd excretion via the mucus plays a substantial role in this regulation, in addition to Cd excretion via the faeces. Snail shells were no sinks for Cd. PMID- 16040173 TI - Potential application of synchronous fluorescence spectroscopy to determine benzo[a]pyrene in soil extracts. AB - Benzo[a]pyrene (BaP) is a significant environmental pollutant and rapid, accurate methods to quantify this compound in soil for both research and environmental investigation purposes are required. In this work, solvent extracts from five contrasting soils spiked with four different polycyclic aromatic hydrocarbons (PAHs) were rapidly analysed by using a synchronous fluorescence spectroscopy (SFS) method. The SFS method was validated using HPLC with ultraviolet detection. A good correlation for the quantification of BaP in soil extracts by the two methods was observed. The detection limit of the SFS method was 1.6 x 10(-9) g/ml in CTAB micellar medium (7.8 mmol/l). The work demonstrates that SFS has potential as a sensitive, accurate, rapid, simple and economic methodology and an efficient alternative to HPLC for fast confirmation and quantification of BaP in complex soil extracts. PMID- 16040174 TI - The combined use of chemical and biochemical markers to assess water quality along the Ebro River. AB - Carp (Cyprinus carpio) and barbel (Barbus graellsii) were collected from five sites along the Ebro River. The study was designed to assess levels of persistent organic pollutants and metals bioaccumulated by fish, and some biochemical responses (cytochrome P450 system, phase II activities, and metallothioneins) against those pollutants. The highest levels of PCBs and DDTs were detected in carp from industrialised areas, which also showed high levels of mercury and cadmium in the liver, and high levels of nonylphenol in bile. Significant alterations in some biochemical markers were observed and associated to combined exposure to pollutants. The activity 7-ethoxyresorufin O-deethylase ranged from 69 pmol/min/mg protein in carp from the reference site to 415 pmol/min/mg protein in those from polluted sites. Carp from the Ebro Delta, an agricultural area, had depressed acetylcholinesterase in muscle tissue. Metallothionein concentrations were positively correlated with copper residues, but no significant differences among sampling points were observed. PMID- 16040175 TI - Determinants of child morbidity in Latin America: a pooled analysis of interactions between parental education and economic status. AB - Diarrhea and respiratory infections account for more than two-fifths of all deaths among children under five. Parental education and economic status are well known risk factors for child morbidity, but little is known about whether education and economic status operate synergistically or independently to influence children's health. Confirming the presence and direction of such interactions is important to better target education and development policies. Our objective is to test for interactions between parental education and economic status in predicting the risk of diarrhea and respiratory illness among children under five, before and after adjusting for key proximate risk factors. We pool 12 Demographic and Health Surveys (DHS) and nine Living Standards Measurement Surveys (LSMS) from Latin America, creating two large databases. Quintiles of economic status are constructed from principal components asset indices. We use logistic regression to analyze episodes of diarrhea and respiratory illness, and interactions between economic quintile and maternal and paternal education are evaluated via likelihood ratio tests. We find that mother's education and quintile interact synergistically in the DHS data, while results are inconclusive in the LSMS data. The effect of increasing maternal education appears to be more protective for children in wealthy families than for children in poor families. Conversely, improvements in economic status reduce health risks more for children whose mothers are better educated. Father's education is protective and operates independently of economic status. Our findings imply that poverty alleviation efforts occurring in concert with programs to educate women and girls will be more effective for improving children's health than either approach alone. PMID- 16040176 TI - Viagra's rise above women's health issues: an analysis of the social and political influences on drug approvals in the United States and Japan. AB - In the United States, Viagra was approved in less than 6 months of its application to the Food and Drug Administration, while the medical abortion pill was approved 4 years after its application, and 17 years after research was first permitted. Congruently, the Ministry of Health in Japan legalized Viagra in 6 months, while oral contraceptives were approved 35 years after the ministry received initial applications. The pharmaceutical review agencies in each country are founded on safety and efficacy standards, in which objective decisions arise from science and clinical investigations. Analyses of these recent drug approvals demonstrate that conclusions may not have been based simply on science and health concerns. Instead, agency actions and application of pharmaceutical law appear to have been influenced by social and political pressures surrounding the products under scrutiny. Pharmaceutical regulations were effectively ignored or manipulated in the United States during the review process for medical abortion, and were applied inconsistently in Japan--ultimately yielding results that happened to conform to contemporary sociopolitical beliefs. Such disregard of legislation holds serious ramifications for public health, national consumer trust and the pharmaceutical industry. It is imperative that external pressures remain outside the scope of drug approval processes. PMID- 16040177 TI - Anti-aging medicine: a patient/practitioner movement to redefine aging. AB - Having enjoyed tremendous growth for the past 5 years, the anti-aging medicine movement is redefining aging so that it becomes a target for biomedical intervention. Targeting aging for intervention dislodges popular understandings of aging: for anti-aging practitioners it no longer matters if aging is natural since it can be itself the target of therapy. So-called "age-associated" diseases like cancer are, in this framework, conceived of as symptoms of aging. Anti-aging medicine is a broad term that may comprise groups selling remedies over the Internet, companies touting the "anti-aging"ness of their products, practitioners who work outside of scientific medicine, and practitioners of anti-aging medicine in clinics who believe that their work is strictly scientific. This article, drawing from more than 3 years of ethnographic interviews, participant observation in clinics and conferences, and a review of the literature, considers the last group. It examines the involvement stories of anti-aging medicine practitioners in two Western United States metropolitan cities. These stories reflect the practices of anti-aging medicine practitioners and the accompanying rationale for involvement. Often originally patients themselves, practitioners frame their involvement with the anti-aging movement in three ways. First, they describe aging as it is currently experienced as a time of decline, suffering, and weakness. This anguish is not inevitable, they argue, and their work toward treating aging biomedically is situated as clearly moral. Secondly, intense frustration with the current biomedical environment has motivated practitioners to look for other ways in which to practice: anti-aging medicine is their chosen alternative. Finally, with dramatic expectations of future biotechnologies and disdain for current medical treatments of old age, anti-aging practitioners embrace a scientific revolutionary identity. These stories of migrations from patient to practitioner reveal the values upon which this movement is grounded and how coming to be a part of it is as much about the movement's mission as it is the origins of the migrations. PMID- 16040178 TI - Salience of self-identity roles in persons with dementia: differences in perceptions among elderly persons, family members and caregivers. AB - In this study, we explored perceptions of the salience of self-identity in persons suffering from dementia as perceived by the participants themselves, by family, and by staff caregivers. Four types of role-identity were explored: professional, family role, hobbies/leisure activities, and personal attributes. Participants were 104 persons with dementia, 48 of whom attended six adult day care centers while 56 resided in two nursing homes in the Washington, DC metropolitan area. Participants, relatives, and staff members were interviewed to obtain information about past and present self-identity roles of participants and attitudes toward these roles. Findings demonstrate that the importance of role identities decreases over time and with the progression of dementia. The family role was found to be the most important and salient role identity according to all the informant groups. The professional role was the one that showed the steepest decline in importance from past to present. Gender differences were detected for the importance of professional role identity. Participants rated their roles in the past as less important and those in the present as more important compared to family members. Family members reported greater decline in the importance of role identities for those participants with greater cognitive impairment. Participants with moderate cognitive impairment reported greater decline in the importance of role identities than did the participants with severe cognitive impairment. Understanding the past and present self-identities of persons with diminished cognitive abilities is crucial in the effort to provide individualized care and enhance participant experiences. PMID- 16040179 TI - IQ decline following early unilateral brain injury: a longitudinal study. AB - We examine whether children with early unilateral brain injury show an IQ decline over the course of development. Fifteen brain injured children were administered an IQ test once before age 7 and again several years later. Post-7 IQ scores were significantly lower than pre-7 IQ scores. In addition, pre-7 IQ scores were lower for children with larger lesions, but children with smaller lesions and higher pre-7 IQ scores showed a greater IQ decline over time. These findings suggest that the cognitive outcomes of children with early lesions, particularly those with relatively small lesions, change over the course of development. PMID- 16040180 TI - Time course of dopamine D2 receptor occupancy by clozapine with medium and high plasma concentrations. AB - Most antipsychotics were thought to induce antipsychotic action at an excess of 70% striatal dopamine D2 receptor occupancy, while the clinical dose of clozapine was reported to show less than 60% occupancy. High-dose clozapine could occupy as high as 80% of striatal dopamine D2 receptor in monkey PET studies. Although the time course of dopamine D2 receptor occupancy is an important property of antipsychotics, that by clozapine has not been investigated in a clinical setting. We measured the time course of extrastriatal dopamine D2 receptor occupancy with different doses of clozapine and evaluated whether the measured occupancies fitted the binding theory. Three consecutive PET scans with [11C]FLB 457 were performed for two patients with schizophrenia, chronically taking 600 mg/day and 200 mg/day of clozapine, respectively. Series of occupancies were also measured in combination with fluvoxamine or paroxetine in one patient. Dopamine D2 receptor occupancies were also simulated using individual clozapine plasma data and previously determined in vivo ED50 value. The occupancy of one patient with high plasma concentration (1207 ng/ml at peak time) was around 75% at peak and around 60% after 26 h. Another patient with medium plasma concentration (649 ng/ml at peak time) showed less than 50% occupancy at peak, decreasing to 15% after 25 h. The measured occupancy values fitted well with the simulated occupancy values. At high plasma concentration, clozapine can induce high extrastriatal dopamine D2 receptor occupancy in the human brain, and this finding fitted well with the theoretical estimation. PMID- 16040181 TI - Mutagenicity of chromium picolinate and its components in Salmonella typhimurium and L5178Y mouse lymphoma cells. AB - Chromium picolinate is one of the most commonly used chromium dietary supplements available in the United States, and it has been marketed to consumers for use in weight loss, increasing muscle mass, and lowering serum cholesterol. Chromium picolinate is a synthetic compound that provides a bioavailable form of Cr(III) that is absorbed better than dietary chromium. However, there are several reports that it can have adverse effects. In order to study the mechanism of observed cellular toxicity and mutagenicity, chromium picolinate and its component compounds, chromium (III) chloride and picolinic acid, were evaluated in Salmonella typhimurium and L5178Y mouse lymphoma cells. Neither chromium picolinate nor chromium chloride induced a mutagenic response in S. typhimurium. However, in the L5178Y mouse lymphoma mutation assay, chromium picolinate induced mutagenic responses without and with the addition of S9. PMID- 16040182 TI - Guidance on setting of acute reference dose (ARfD) for pesticides. AB - This paper summarises and extends the work developed over the last decade by the Joint FAO/WHO Meeting on Pesticide Residues (JMPR) for acute health risk assessment of agricultural pesticides. The general considerations in setting of acute reference doses (ARfDs) in a step-wise process, as well as specific considerations and guidance regarding selected toxicological endpoints are described in detail. The endpoints selected are based on the practical experience with agricultural pesticides by the JMPR and are not a comprehensive listing of all possible relevant endpoints. Haematotoxicity, immunotoxicity, neurotoxicity, liver and kidney toxicity, endocrine effects as well as developmental effects are taken into account as acute toxic alerts, relevant for the consideration of ARfDs for pesticides. The general biological background and the data available through standard toxicological testing for regulatory purposes, interpretation of the data, conclusions and recommendations for future improvements are described for each relevant endpoint. The paper also considers a single dose study protocol. This type of study is not intended to be included in routine toxicological testing for regulatory purposes, but rather to guide further testing when the current database indicates the necessity for an ARfD but does not allow a reliable derivation of the value. PMID- 16040183 TI - Effects of oximes on muscle force and acetylcholinesterase activity in isolated mouse hemidiaphragms exposed to paraoxon. AB - Toxicity of organophosphates (OP) is caused by inhibition of acetylcholinesterase (AChE), resulting in accumulation of acetylcholine. While cholinolytics such as atropine are able to counteract muscarinic symptoms, they are unable to restore the impaired neuromuscular transmission (NMT). Here, oximes as potential reactivators of inhibited AChE may be effective. Until now, no unequivocal relation between oxime-induced increase in muscle force and reactivation has been demonstrated. To address this issue the isolated circumfused mouse hemidiaphragm was used as an experimental model. The muscle force generation upon tetanic stimuli was recorded during AChE inhibition by 1 microM paraoxon and after a wash out period in the presence of obidoxime, pralidoxime and the experimental oximes HI 6, and HLo 7, 10 microM each. At the end of the experiments AChE activity was determined in the diaphragm homogenates by a radiometric assay. At 50-Hz stimulation, recovery was complete with obidoxime, nearly complete with HLo 7 but incomplete with HI 6 and pralidoxime. Only with obidoxime a significant increase in AChE activity was found. An increase of AChE to 10% of normal was sufficient to allow normal muscle force generation. When paraoxon was still present, obidoxime and HLo 7 were effective at 0.1 microM paraoxon, but failed so at paraoxon >1 microM. The data show different effectiveness of the oximes investigated in reactivation of muscle AChE and recovery of NMT after inhibition by paraoxon. Although an increase in muscle force by the oximes was accompanied by a measurable increase in AChE activity only in the case of obidoxime, the plot of muscle force against AChE activity as well as lacking evidence for a direct effect and adaptive processes indicate that reactivation of the enzyme is the main mechanism of NMT recovery. In agreement, in presence of AChE inhibitory concentrations of paraoxon during reactivation a reduced effectiveness of oximes was found. PMID- 16040184 TI - L-Arginine modulates CXC chemokines in the human intestinal epithelial cell line HCT-8 by the NO pathway. AB - Arginine has immunomodulating properties in different animal models but its effects in human intestine remain unknown. This study examined whether arginine modulates inflammatory mediators as chemokines and nitric oxide (NO) in the human intestinal epithelial cell line HCT-8 induced by cytokines. Under basal conditions, arginine did not influence iNOS protein expression, NO and chemokine production and mRNA levels (P>0.05 for all). Stimulation with cytokines-induced a significant increase of NO and chemokine production, iNOS and chemokine mRNA level and iNOS protein expression. Under inflammatory conditions, arginine increased 30% NO production (P<0.05) but did not influence iNOS mRNA level or iNOS protein expression. Under stimulated conditions, arginine decreased IL-8 and Mig mRNA level (57% and 39%, for 0.1 vs. 2 mmol/l l-arginine, P<0.05, respectively), and production (respectively, 28 and 23%, both P<0.05). IP-10 and I-TAC mRNA level and production were not significantly influenced by arginine. Under inflammatory conditions, l-arginine as well as a NO donor (sodium nitroprusside (SNP)) increased NO production, which was inversely correlated with IL-8 production (r'=-0.66, P=0.007 for arginine; r'=-0.79, P<0.0001 for SNP). Use of NG-Methyl-l-arginine acetate, a NOS inhibitor which prevents arginine-induced NO production, suppressed the arginine-induced IL-8 inhibition (P<0.05). In HCT-8 cells, arginine enhanced cytokine-induced NO production, reduced IL-8 and Mig production and mRNA level and had no effects on other assessed chemokines. In conclusion, arginine-induced IL-8 inhibition in HCT-8 cells involves NO pathway under inflammatory conditions. These data suggest that arginine-enriched enteral nutrition may have significant influence on inflammatory response in human intestine. PMID- 16040185 TI - Biochemical properties of cytochrome c nitrated by peroxynitrite. AB - Nitration of tyrosine residues is taken as evidence for intracellular formation of peroxynitrite. Cytochrome c (cyt c) can be nitrated by peroxynitrite and nitrated cyt c has been observed in cells and tissues under stress conditions. Here we studied the biochemical properties of nitrated cyt c in order to understand its potential roles in nitrative stress. Nitration of cyt c resulted in disruption of the heme-methionine bond and rapid binding to cyanide. Equilibrium unfolding by guanidine hydrochloride showed that cyt c was slightly destabilized upon nitration but the unfolding transition of nitrated cyt c was highly cooperative indicating that the overall folding was largely preserved. Nitrated cyt c could not be reduced by superoxide and did not support electron transfer between ascorbate and cyt c oxidase. Nitration of cyt c resulted in a tremendous increase in peroxidase activity so that nitrated cyt c rapidly oxidized dihydrodichlorofluorescein even in the presence of a high concentration of glutathione. Enhanced peroxidase activity of nitrated cyt c was responsible for H2O2-induced oxidation of phospholipid membranes and H2O2/NO2--mediated nitration of other proteins. These results suggest that nitration of cyt c by peroxynitrite may exacerbate oxidative damage to mitochondrial proteins and membranes. PMID- 16040186 TI - Response of the anaerobe Desulfovibrio vulgaris Hildenborough to oxidative conditions: proteome and transcript analysis. AB - The method of two-dimensional protein gel electrophoresis was used to evaluate the changes at the proteins level following oxygen exposure of the anaerobic sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough. Fifty-seven proteins showed significant differential expression. The cellular concentration of 35 proteins decreased while that of nineteen increased as a specific consequence of oxidative conditions. The proteins that were less abundant belonged to various functional categories such as nucleic acid and protein biosynthesis, detoxification mechanisms, or cell division. Interestingly, quantitative real-time PCR revealed that the genes encoding detoxification enzymes (rubrerythrins, superoxide reductase) are down regulated. The loss of viability of D. vulgaris Hildenborough under these oxidative conditions (Fournier et al., J. Biol. Chem. 279 (2004) 1785) can be directly related to the decrease in the cellular concentrations of these proteins, thereby specifying the toxicity of oxygen for the cells. Among the proteins that were more abundant under oxygen exposure, several thiol-specific peroxidases (thiol-peroxidase, BCP-like protein, and putative glutaredoxin) were identified. Using RT-PCR, the up-regulation of the genes encoding the thiol-peroxidase and the BCP was demonstrated. That is the first time that these proteins have been shown to be involved in the defense of D. vulgaris toward an oxidative stress. Several hypothetical proteins were also shown to be differentially expressed. A function in the defense mechanism against an oxidative stress is proposed for these uncharacterized proteins. PMID- 16040187 TI - Expression and intracellular localization of Pyk2 in normal and v-src transformed chicken epiphyseal chondrocytes. AB - The expression and localization of prolin-rich tyrosine kinase 2 (Pyk2) were studied in chick embryo epiphyseal chondrocytes. Two immunoreactive bands were detected in chondrocytes, a major band with an apparent Mr of 123 kDa and a minor band with an apparent Mr of 68 kDa. The major band appears to migrate as a doublet with apparent Mr of 116/123 kDa. Increased levels of the three forms of Pyk2 were observed in v-src transformed chondrocytes as compared to control uninfected chondrocytes. Immunofluorescent staining shows that Pyk2 is clearly visible in the cytosol and in the perinuclear region of control and v-src chondrocytes and displays a pattern very similar to the distribution of the mitochondrial marker Mito Tracker. More, immunofluorescent staining shows that Pyk2 is nuclear in most chondrocytes. By subcellular fractionation, the p116/123 Pyk2 doublet, was found to be accumulated mainly in the cytoplasm while the p68 Pyk2 form, was found to be accumulated exclusively in the nucleus. The differential nuclear/cytoplasmic distribution of the Pyk2 forms remains unchanged after v-Src-induced transformation. The p68 Pyk2 form could no longer be detected by using a N-terminus domain-specific anti-Pyk2 antibody. Consistently, Pyk2 immunoreactivity was restricted to the cytoplasm of control and v-src transformed chondrocytes. Thus it appears that the p68 Pyk2 form that accumulates in the nucleus has a deletion in the N-terminus region. PMID- 16040189 TI - Are neurology residents in the United States being taught defensive medicine? AB - OBJECTIVE: To study whether and how fear of litigation and defensive medicine are communicated during residency training and to assess whether this affects residents' attitudes. METHODS: Neurology residents in the US (n=25) and, as a control group, Neurology residents training in Germany (n=42) were asked to rate multiple items regarding litigation, defensive strategies and how often these issues are raised by teaching physicians. Statistic analysis was performed using nonparametric tests. RESULTS: Residents in both countries indicated that litigation is an "important problem", although US residents stated this significantly more often (p<0.001). Initiation of tests motivated mainly by fear of litigation (p=0.004) and explicit teaching of defensive strategies by teaching physicians (p<0.02) were reported more often by US residents. CONCLUSION: Neurology residents in both the US and Germany perceive a litigational threat, but significantly less so in Germany. This difference may result at least in part from teaching of defensive strategies reported more often in US programs. PMID- 16040188 TI - Transport of N-acetyl-D-galactosamine in Escherichia coli K92: effect on acetyl amino sugar metabolism and polysialic acid production. AB - The N-acetyl-D-galactosamine (GalNAc) transport system of Escherichia coli K92 was studied when the bacterium was grown in a chemically defined medium containing GalNAc as a carbon source. Kinetic measurements were carried out in vivo at 37 degrees C in 25 mM phosphate buffer, pH 7.0. Under these conditions, the uptake rate was linear for at least 3 min and the calculated Km for GalNAc was 3 microM. The transport system was strongly inhibited by sodium arsenate (70%), potassium cyanide (62%) and 2,4-dinitrophenol (75%). Analysis of bacterial GalNAc phosphotransferase activity revealed in vitro GalNAc phosphorylation activity only when phosphoenolpyruvate was present. These results strongly support the notion that GalNAc uptake depends on a specific phosphotransferase system. Study of activity regulation showed that N-acetylglucosamine and mannosamine specifically inhibit the transport of GalNAc in this bacterium. Analysis of expression revealed that the GalNAc transport system is specifically induced by GalNAc but not by N-acetylglucosamine (GlcNAc) or N-acetylmannosamine (ManNAc), two intimately related sugars. Moreover, full induction of GalNAc transport required the presence of both cAMP and GalNAc. Comparative studies revealed that E. coli K92 has developed a regulation mechanism that specifically induces the appropriate permease based on the presence of each respective phospho amino sugar (GlcNAc, ManNAc and GalNAc). In this regulation system, GlcNAc is the preferred amino sugar as the carbon source. Finally, when E. coli K92 was grown using GalNAc, capsular polysialic acid production was strongly affected. The presence of intracellular phosphoderivative acetylamino sugars, generated by the action of the phosphotransferase transport system, can be responsible for this effect. PMID- 16040190 TI - Adsorption of Bismark Brown dye on activated carbons prepared from rubberwood sawdust (Hevea brasiliensis) using different activation methods. AB - Hevea brasiliensis or rubberwood tree, as it is commonly known finds limited use once the latex has been tapped. The sawdust of this tree is chosen to ascertain it viability as a precursor for activation. The carbons thus obtained were characterized in terms of iodine, methylene blue number and surface area. The best carbon in each method was utilized to study the adsorption of Bismark Brown, a dye used in the leather industry. Adsorption equilibrium studies were carried out with the synthetic solutions of the dye, at room temperature (298 K). Equilibrium data are fitted with the Langmuir and the Freundlich isotherms models for the system. The effects of contact time, adsorbent dosage and initial dye concentrations on sorption capacity were carried out. Excellent adsorption capacities of 2000 and 1111 mg g(-1) were obtained for steam and chemical followed by steam-activated carbons, respectively. Pilot-plant experimental studies have been performed using packed-bed column with different feed concentrations, flow rates and bed heights, to evaluate sorption of Bismark Brown on steam-activated carbon. Bed depth service time (BDST) design model have been used to analyze the data. PMID- 16040191 TI - Allele epsilon 4 of APOE is a stronger predictor of Alzheimer risk in Sicily than in continental South Italy. AB - The genotype of apolipoprotein E was examined in 173 sporadic Alzheimer's disease (AD) patients, 132 with late onset (LOAD) and 41 with early onset (EOAD), and in 174 healthy matched controls from Sicily. Despite a low frequency of the epsilon 4 allele (6.3%, 95% CI: 4.2--9.4) in controls, epsilon 4 allele was a stronger predictor of AD risk (odds ratio: 5.8, 95% CI: 3.5--9.4; p<0.0001) than in most of the studies performed in other regions of Italy, and it has no influence on age at onset. epsilon 4/epsilon 4 and epsilon 4/epsilon 3 genotypes were similar predictors of AD risk. Conversely, a decreased risk was found in epsilon 3 allele carriers (odds ratio: 0.3, 95% CI: 0.2--0.4; p<0.0001), which remained significant when considering EOAD cases only (odds ratio: 0.2, 95% CI: 0.1--0.4, p<0.0001). In conclusion, differences in association strength of epsilon 4 allele with AD between Sicily and other regions of Italy suggest an influence of complex gene-gene and gene-environment interactions. PMID- 16040192 TI - Theta and alpha oscillations reflect differences in memory strategy and visual discrimination performance in patients with Parkinson's disease. AB - The aim of the present study was to investigate alterations in brain oscillatory activity during transient intentional encoding of abstract novel shapes in patients with Parkinson's disease (PD). Combined time-frequency analyses of alpha and theta power revealed a clear dissociation between PD patients (N=10) and age matched healthy controls (N=10). PD patients used sub-optimal stimulus encoding strategies for subsequent maintenance and recall, whereas the controls mainly used categorization processes. In contrast to controls, PD patients showed significantly less theta increase and upper alpha suppression at frontal locations and significant laterality of early posterior theta and lower-2 alpha oscillations at right posterior locations. These findings suggest that the fronto striatal circuits participate in visual categorization processes. Furthermore, theta and alpha oscillations appear to be involved in mediating the integration processes in mnemonic networks underlying visual categorization. PMID- 16040193 TI - Visual deprivation effects on human motor cortex excitability. AB - Single and paired-pulse transcranial magnetic stimulation (TMS) were applied to the motor cortex of 12 healthy volunteers, who were instructed to relax under eyes-open and eyes-closed conditions with room lights on and after 30 min of blindfolding. Compared to the eyes-open condition, significantly larger motor evoked potentials and less intracortical inhibition were observed during blindfolding. Visual deafferentation changes resting human motor cortex excitability and might be a novel way to promote brain plasticity. These results raise the issue of how widespread the effects of temporary deafferentation may be and whether they are mediated by discrete or diffuse systems. These findings also illustrate an important potential confound in TMS studies of the motor cortex. PMID- 16040194 TI - The transmethylation cycle in the brain of Alzheimer patients. AB - Homocysteine accumulation, frequently observed in plasma of AD patients, may be a sign of a reduced activity of the brain methionine-homocysteine transmethylation cycle. S-Adenosylmethionine (SAM) is the main methyl donor in several transmethylation reactions. The demethylated product of SAM, S adenosylhomocysteine (SAH), is hydrolyzed to yield homocysteine, which can be remethylated to methionine by transfer of a methyl group of 5 methyltetrahydrofolate (5-MTHF). A reduced activity of the transmethylation cycle in the brain may result in hypomethylation of the promoter of the presenilin 1 (PS1) gene, which will lead to overexpression of presenilin 1 and, consequently, to increased Abeta(1-42) (Abeta42) formation. Brain transmethylation was studied in 30 patients with 'probable' AD and 28 age-matched non-demented controls by measuring the cerebrospinal fluid (CSF) levels of SAM, SAH and 5-MTHF. 5-MTHF was determined by HPLC with electrochemical detection, while SAM and SAH were assayed by stable isotope dilution tandem mass spectrometry. We found no statistical differences between AD patients and controls for 5-MTHF, SAM and SAH levels, and the SAM/SAH-ratio in CSF. These findings argue against a possible change in methylation of the promoter and expression of PS1. PMID- 16040195 TI - Words can help! PMID- 16040196 TI - Letter to the editor regarding Morley et al. Possible selves in chronic pain: self-pain enmeshment, adjustment and acceptance. PMID- 16040198 TI - Hidden Markov model-derived structural alphabet for proteins: the learning of protein local shapes captures sequence specificity. AB - Understanding and predicting protein structures depend on the complexity and the accuracy of the models used to represent them. We have recently set up a Hidden Markov Model to optimally compress protein three-dimensional conformations into a one-dimensional series of letters of a structural alphabet. Such a model learns simultaneously the shape of representative structural letters describing the local conformation and the logic of their connections, i.e. the transition matrix between the letters. Here, we move one step further and report some evidence that such a model of protein local architecture also captures some accurate amino acid features. All the letters have specific and distinct amino acid distributions. Moreover, we show that words of amino acids can have significant propensities for some letters. Perspectives point towards the prediction of the series of letters describing the structure of a protein from its amino acid sequence. PMID- 16040199 TI - The addition of tretinoin to permethrin, pyrethrins, and DDT for production of powerful anti-lice agents. PMID- 16040200 TI - Impaired fatty acid oxidation as a cause of liver disease associated with hyperemesis gravidarum. AB - Hyperemesis gravidarum (HG) is the most severe form of illness within the spectrum of nausea and vomiting of pregnancy. Liver disease, usually consisting of mild serum transaminase elevation, occurs in almost 50% of patients with HG. While multiple risk factors have been proposed, the etiology and underlying mechanism of maternal liver disease associated with HG remains unclear. In this report, we hypothesize that impairment of mitochondrial fatty acid oxidation (FAO) plays a role in the pathogenesis of maternal liver disease associated with HG. We hypothesize that women heterozygous for FAO defects develop HG associated with liver disease while carrying fetuses with FAO defects due to accumulation of fatty acids in placenta and subsequent generation of reactive oxygen species. Alternatively, it is possible that starvation leading to peripheral lipolysis and increased load of fatty acids in maternal-fetal circulation, combined with reduced capacity of the mitochondria to oxidize fatty acids in mothers heterozygous for FAO defects, can also cause HG and liver injury while carrying non-affected fetuses. The rationale for this hypothesis is discussed. PMID- 16040201 TI - Medical and nonmedical use of prescription pain medication by youth in a Detroit area public school district. AB - The purpose of this descriptive study was to examine the nonmedical use of prescription pain medications in a sample of public school students in the Detroit metropolitan area. The Web-based survey occurred during a two-week period in May 2003 and drew on a diverse population between the ages of 10 and 18 years. An 87% response rate was achieved, providing a final sample of 1017 students. Twenty-two percent of girls and 10% of boys reported lifetime nonmedical use of a pain medication (chi2 = 25.2, d.f. = 1, p < 0.001). Fifteen percent of girls and 7% of boys reported past year nonmedical use (chi2 = 12.7, d.f. = 1, p < 0.01). Compared to nonusers, nonmedical users of prescription pain medication were seven times more likely to smoke cigarettes, five times more likely to drink alcohol and smoke marijuana, almost four times more likely to binge drink, and eight times more likely to have used other illicit drugs. Of those reporting lifetime nonmedical use of prescription pain medication, 70% noted how they obtained the drugs; 34% reported getting the drugs from a family member, 17% from a friend and 14% from a dealer or theft. PMID- 16040202 TI - The nematode Panagrellus redivivus is susceptible to killing by human pathogens at 37 degrees C. AB - Caenorhabditis elegans has been used as a host for the study of bacteria that cause disease in mammals. However, a significant limitation of the model is that C. elegans is not viable at 37 degrees C. We report that the gonochoristic nematode Panagrellus redivivus survives at 37 degrees C and maintains its life cycle at temperatures up to and including 31.5 degrees C. The C. elegans pathogens Pseudomonas aeruginosa, Salmonella enterica, Staphylococcus aureus, but not Yersinia pseudotuberculosis, reduced P. redivivus lifespan. Of four strains of Burkholderia multivorans tested, one reduced P. redivivus lifespan at both temperatures, one was avirulent at both temperatures and two strains reduced P. redivivus lifespan only at 37 degrees C. The mechanism by which one of these strains killed P. redivivus at 37 degrees C, but not at 25 degrees C, was investigated further. Killing required viable bacteria, did not involve bacterial invasion of tissues, is unlikely to be due to a diffusible, bacterial toxin and was not associated with increased numbers of live bacteria within the intestine of the worm. We believe B. multivorans may kill P. redivivus by a temperature regulated mechanism similar to B. pseudomallei killing of C. elegans. PMID- 16040203 TI - Conidial anastomosis tubes in filamentous fungi. AB - Conidial anastomosis tubes (CATs) can be recognized in 73 species of filamentous fungi covering 21 genera, and develop in culture and in host-pathogen systems. They have been shown to be morphologically and physiologically distinct from germ tubes in Colletotrichum and Neurospora, and under separate genetic control in Neurospora. CATs are short, thin, usually unbranched and arise from conidia or germ tubes. Their formation is conidium-density dependent, and CATs grow towards each other. MAP kinase mutants of Neurospora are blocked in CAT induction. Nuclei pass through fused CATs and are potential agents of gene exchange between individuals of the same and different species. CAT fusion may also serve to improve the chances of colony establishment. PMID- 16040204 TI - The yicM (nepI) gene of Escherichia coli encodes a major facilitator superfamily protein involved in efflux of purine ribonucleosides. AB - The yicM gene of Escherichia coli was found by selection for resistance to 6 mercaptopurine. Translation and transcription initiation sites of yicM were determined. Overexpression of yicM increased resistance of sensitive cells to inosine and guanosine, decreased E. coli growth rate in medium containing these ribonucleosides as the sole carbon source, led to inosine accumulation by the E. coli strain deficient in purine nucleoside phosphorylase and enhanced the rate of inosine excretion by an inosine-producing strain. These results suggest that yicM encodes a purine ribonucleoside exporter and we have accordingly renamed it nepI (for 'nucleoside efflux permease-inosine'). PMID- 16040205 TI - Bioluminescence-mediated stimulation of photoreactivation in bacteria. AB - Although biochemistry and genetics of light emission by cells have been investigated in detail, a biological role for bacterial luminescence has remained obscure for a long time. It was proposed recently that luminescence may stimulate DNA repair, but the specific mechanism of this phenomenon was not investigated. Moreover, experiments showing decreased survival of UV-irradiated lux mutants relative to luminescent cells were performed previously using only one bacterial species, Vibrio harveyi. Here, we demonstrate that dark mutants of various strains of naturally luminescent bacteria (Photobacterium leiognathi, Photobacterium phosphoreum and Vibrio fischeri) are more sensitive to UV irradiation than wild-type cells. Thus, this phenomenon occurs not only in V. harveyi but also in other bacterial species. Using an artificial system of luminescent Escherichia coli in combination with phr mutants (defective in photolyase functions), we found that bacterial luminescence may stimulate photoreactivation, perhaps by providing photons that are necessary for photolyase activity. PMID- 16040206 TI - Genetic evidence that mycobacterial FtsZ and FtsW proteins interact, and colocalize to the division site in Mycobacterium smegmatis. AB - We provide genetic evidence to show that the Mycobacterium tuberculosis FtsZ and FtsW proteins interact, and that these interactions are biologically relevant. Furthermore, we show by fluorescence microscopy that Mycobacterium smegmatis FtsW is part of its septasomal complex and colocalizes with FtsZ to the midcell sites. Colocalization experiments reveal that approximately 27% of the cells with septal Z-rings contain FtsW whereas 93% of the cells with FtsW bands are associated with FtsZ indicating that FtsW is late recruit to the septum, as in Escherichia coli. Our results suggest that mycobacterial FtsZ can localize to the septum independent of FtsW, and that interactions of FtsW with FtsZ are critical for the formation of productive FtsZ-rings and the cell division process in mycobacteria. PMID- 16040207 TI - Downregulation of TACO gene transcription restricts mycobacterial entry/survival within human macrophages. AB - Recent reports have indicated that cholesterol-dependent association of tryptophan-aspartate containing coat protein (TACO) plays a crucial role in the entry/survival of Mycobacterium tuberculosis within human macrophages. Keeping this in view, the present study explored whether the molecules that have the ability to downregulate TACO gene transcription could also restrict entry/survival of mycobacteria within human macrophages. The study revealed that chenodeoxycholic acid (CDCA), either alone or in combination with retinoic acid (RA), had the inherent capacity to downregulate TACO gene transcription in a dose dependent fashion. This result was in conformity with the existence of a functional FXR/RXR binding site analyzed in the regulatory region of the TACO gene. Furthermore, we demonstrate that the entry and intracellular survival of M. tuberculosis is significantly restricted in THP-1 macrophages exposed to CDCA/RA. On the basis of these findings, we propose that the CDCA/RA-dependent pathway may open a new possibility for the treatment of tuberculosis. PMID- 16040208 TI - SEP-class genes in Populus tremuloides and their likely role in reproductive survival of poplar trees. AB - One of the most important processes to the survival of a species is its ability to reproduce. In plants, SEPALLATA-class MADS-box genes have been found to control the development of the inner whorls of flowers. However, while much is known about floral development in herbaceous plants, similar systems in woody trees remain poorly understood. Populus tremuloides (trembling aspen) is a widespread North American tree having important economic value, and its floral development differs from that of well-studied species in that the flowers have only two whorls and are truly unisexual. Sequence based analyses indicate that PTM3 (Populus tremuloides MADS-box 3), and a duplicate gene PTM4, are related to the SEPALLATA1-and 2-class of MADS-box genes. Another gene, PTM6, is related to SEP3, and each of these genes has a counterpart in the poplar genomic database along with additional members of the A, B, C, D, and E-classes of MADS-box genes. PTM3/4 and 6 are expressed in all stages of male and female aspen floral development. However, PTM3/4 is also expressed in the terminal buds, young leaves, and young stems. In situ RNA localization identified PTM3/4 and 6 transcripts predominantly in the inner, sexual whorl, within developing ovules of female flowers and anther primordia of male flowers. Tree researchers often use heterologous systems to help study tree floral development due to the long juvenile periods found in most trees. We found that the participation of PTM3/4 in floral development is supported by transgenic experiments in both P. tremuloides and heterologous systems such as tobacco and Arabidopsis. However, phenotypic artifacts were observed in the heterologous systems. Together the results suggest a role for poplar SEP-class genes in reproductive viability. PMID- 16040209 TI - Myocardial infarction risk and hormone replacement: differences between products. AB - OBJECTIVES: To establish the risk of myocardial infarction (MI) in users of hormone replacement therapy (HRT) compared with non-users and to compare the risk between different HRT regimens. METHODS: A population-based cohort and case control study, and a case-control study nested within a cohort of HRT users, using the UK General Practice Research Database. Differences between HRT regimen, mode of administration and duration and recency of use were examined whilst adjusting for confounding. RESULTS: In the cohort and case-control study, 4537 cases of MI were identified in 2.62 million observed women years, cases were age matched to 27,220 controls. In both studies, current and past HRT use were associated with reduced risk estimates for MI compared with no prior use. MIs were less likely to be fatal amongst women who had used HRT than amongst never users (OR(adj) 0.58; 95% CI 0.45-0.75). No difference in risk was seen between current and past use, oral and transdermal HRT or between different regimens (p>0.44). In the nested study, no difference was found in the association with MI risk between different oestrogen-progestogen combinations or between different combinations and tibolone. Unopposed oestrogen use was not associated with a decrease in risk compared with combined HRT. CONCLUSIONS: These results are consistent with previous observational studies in supporting the hypothesis that use of postmenopausal HRT is associated with a decrease in risk of acute myocardial infarction (AMI). Case fatality differed between HRT users and non users, suggesting a protective effect of HRT. This study does not demonstrate a difference between regimens. PMID- 16040210 TI - The effect of intranasal 17 beta-estradiol on mammographic breast density. PMID- 16040211 TI - Conjugated estrogen plus medroxyprogesterone does not impair blood rheological properties in hypertensive postmenopausal women. AB - OBJECTIVES: Hypertension and estrogens are both prothrombotic. We used the microchannel method to investigate whether hormone replacement therapy (HRT) with conjugated equine estrogen (CEE) plus medroxyprogesterone acetate (MPA) affects blood flow through the microchannels in hypertensive postmenopausal women being treated with antihypertensive drugs and in normotensive postmenopausal women. METHODS: Sixty-two consecutive postmenopausal women were randomly assigned to a hypertensive HRT group (n=16), hypertensive control group (n=15), normotensive HRT group (n=16) and normotensive control group (n=15). Each HRT group received CEE 0.625 mg plus MPA 2.5 mg daily orally for 12 months. Both hypertensive groups were being treated with antihypertensive drugs before the study. Microvascular blood flow was assessed on the basis of blood passage time, the time required for 100 microl of whole blood to pass through a cylinder, was determined before and 12 months after the start of HRT by the microchannel method (micro channel array flow analyzer). RESULTS: CEE plus MPA therapy did not change blood passage time in any of the groups. Microscopic observation showed that the whole blood passed smoothly through the microchannels in every group. CONCLUSIONS: CEE plus MPA therapy may not impair blood flow through the microchannels in hypertensive postmenopausal women receiving antihypertensive drugs or in normotensive postmenopausal women. However, administration of CEE plus MPA to postmenopausal women with hypertension warrants caution against the occurrence of thromboembolic events. PMID- 16040212 TI - Hormone replacement therapy in Germany: determinants and possible health-related outcomes. Results of National Health Surveys from 1984 to 1999. AB - OBJECTIVES: To explore the determinants of hormone replacement therapy (HRT) use and possible health-related outcomes in Germany. METHODS: 827 current HRT users (age range 31-78 years, who had taken HRT products in the last 7 days before the medical interview) and 2371 age-matched nonusers were identified from five population-representative National Health Surveys performed from 1984 to 1999 in Germany. Cross-sectional comparisons between HRT users and nonusers were performed for socioeconomic factors and possible health-related outcomes. Regression models were used to obtain odds ratios of HRT use. RESULTS: HRT use increased dramatically from 3% in 1984/1985 to 21% in 1998/1999 among German women aged 40-69. Regression analysis and comparison between HRT users and nonusers suggested that HRT use was closely associated with socioeconomic factors and personal lifestyle that may favor better health. In HRT users, however, we did not observe any health-related benefits in satisfaction with life or health. While HRT users appeared superior to nonusers with respect to the glycemic status, histories of cardiovascular diseases, despite a favorable lipid profile, occurred not less than in the group of nonusers. CONCLUSIONS: In the settings of our cross-sectional studies in Germany HRT did not improve users' health benefits and did not show cardioprotective effects. The better glycemic status should be balanced against the higher risks of cardiovascular diseases and breast cancer. PMID- 16040213 TI - Controlled release of levonorgestrel from biodegradable poly(D,L-lactide-co glycolide) microspheres: in vitro and in vivo studies. AB - Poly(D,L-lactide-co-glycolide) (PLG) biodegradable microspheres containing a contraceptive drug, levonorgestrel (LNG), were prepared using both the solvent evaporation method and a modified solvent extraction-evaporation method. The microspheres prepared with the solvent evaporation process had porous surfaces with low product yields and poor encapsulation efficiencies. On the other hand, the microspheres prepared using the modified solvent extraction-evaporation method were nonporous with encapsulation efficiencies close to 100%. In vitro drug release showed the nonporous microspheres had a lower initial burst and a slightly prolonged duration of release than those porous microspheres. In vivo release kinetics of the low burst microspheres were determined by measuring LNG plasma levels after a single intramuscular injection to female rats. At a LNG dose of 41.1 mg/kg, average plasma LNG levels were 6-10 ng/ml in the first 24 h and subsequently remained above 1 ng/ml until 126 days. The duration above the minimum effective LNG plasma level of 0.2 ng/ml was 168 days. By comparison, a similar dose of LNG microcrystals used as control produced a much higher plasma level of 15-21 ng/ml in the first day followed by a fast and continuous decline of LNG levels with a duration of only about 35 days. PMID- 16040214 TI - Thermal behaviour and stability in Olanzapine. AB - The stability and thermal behaviour of two anhydrate phases and a new mixed water:DMSO solvate of Olanzapine (2-methyl-4-(4-methyl-1-piperazinyl)-10H-thieno [2,3-b][1,5]benzodiazepine) are studied by different methods: differential scanning calorimetry (DSC), X-ray powder diffraction (XRPD) and Raman scattering (RS). Single crystal structural data for the latter phase are presented, confirming the presence of the (Olanzapine)(2) dimer as the structural building unit of all known phases of the drug, either anhydrate or solvated. An apparent interconversion between both solid state forms is shown to be an artifact and explained in terms of a melting-recrystallization process. PMID- 16040216 TI - Antibacterial and antioxidant activities of ethanol extract from Paullinia cupana Mart. AB - The antibacterial and antioxidant activity of the ethanol extract from Paullinia cupana var. sorbilis Mart. (Sapindaceae) seeds, commonly called guarana, was assessed towards selected bacteria as well as in different antioxidant models. The extract, at a concentration between 16 and 128 microg/ml, showed a significant antibacterial effect expressed as minimum inhibitory concentration (MIC) against both Gram-negative and Gram-positive bacteria. In particular, Pseudomonas aeruginosa (MIC=16 microg/ml), Proteus mirabilis (MIC=32 microg/ml), Proteus vulgaris (MIC=32 microg/ml) and Escherichia coli (MIC=32 microg/ml) were the most inhibited. The antioxidant activity was determined by the malonyldialdehyde (MDA) test, measuring the MDA concentration in 3T3-L1 cells after induced cellular damage using ferric ammonium citrate (FAC). The reduction of lipid peroxidation was 62.5% using a guarana extract with a concentration of 2 microg/ml. This effect was dose/dependent. The ethanol extract from Paullinia cupana seeds was analysed by spectrophotometry to determine the concentration of catechol substances after treatment of the extract with p-aminophenol. The total phenolics content in the ethanol extract was also determined spectrophotometrically according to the Folin-Ciocalteu procedure and calculated as gallic acid equivalents (GAE). The concentration of catechol equivalent was 6.06+/-0.13 mg/g (mean+/-S.D.), while the total phenolic content was 8.43+/-0.21 mg/g (mean+/-S.D.). The correlation index between antioxidant activity and catechol content was 0.96. PMID- 16040217 TI - In vitro tests and ethnopharmacological investigations: wound healing as an example. AB - In vitro tests are now widely employed in ethnopharmacological research because of ethical reasons and their usefulness in bioactive-guided fractionation and determination of active compounds. For many disease conditions, a variety of in vitro tests can now be employed as the biochemical mechanisms underlying disease and healing processes are understood. Approaches to the in vitro investigations of wound healing processes are exemplified by studies on extracts of Buddleja species and three Ghanaian species Spathodea campanulata, Commelina diffusa and Secamone afzelii. Most studies have been carried out on Buddleja officinalis or Buddleja globosa. The extracts have been shown to have anti-inflammatory and antioxidant properties due to flavonoids, triterpenoids, diterpenoids and caffeic acid derivatives. There appears to a slight effect on proliferation of fibroblasts at lower concentrations, but this was not significant, and higher concentrations appeared to be cytotoxic. Novel findings are the ability of Buddleja globosa leaf extracts to induce differentiation in keratinocytes and to alter the profile of proteins produced by cultured fibroblasts. Extracts also had some effect on lattice contraction. The three Ghanaian species examined show a mixture of antimicrobial and antioxidant activities. The evolution over recent years of tests for wound healing, from in vivo tests to cell-based systems and chemical reactions and on to investigations into effects on secondary messengers and protein expression, is described. PMID- 16040218 TI - TNF-alpha and endotoxin increase hypoxia-induced VEGF production by cultured human nasal fibroblasts in synergistic fashion. AB - OBJECTIVE: Vascular endothelial growth factor (VEGF) promotes angiogenesis and is associated with the invasion and metastasis of malignant tumors. It enhances vascular permeability and is expressed in inflammatory nasal as well as middle ear mucosa. As the mechanism of VEGF induction during chronic inflammation, such as chronic paranasal sinusitis (CPS) remains to be clarified, we studied the factors regulating the production of VEGF in cultured human nasal fibroblasts and discussed the role of VEGF in the pathogenesis of CPS. METHODS: We used ELISA to quantify VEGF levels in paranasal sinus effusions, nasal secretions, and serum from patients with CPS. In addition, we cultured human nasal fibroblasts isolated from nasal polyps of CPS patients and studied the effects of hypoxia, TNF-alpha, and endotoxin on their production of VEGF using ELISA and PCR. RESULTS: The VEGF concentration was significantly higher in paranasal sinus effusions than in nasal secretions and serum. Nasal fibroblasts produced high levels of VEGF, when cultured under hypoxic condition and this production was remarkably enhanced in the presence of TNF-alpha or endotoxin. CONCLUSION: VEGF is locally produced in paranasal sinuses as well as nasal mucosa and its production is increased in patients with CPS. Hypoxia is associated with the production of VEGF by nasal fibroblasts and TNF-alpha and endotoxin may act synergistically to enhance VEGF production in paranasal sinuses under hypoxic condition. PMID- 16040219 TI - Tolerability of N-chlorotaurine in chronic rhinosinusitis applied via yamik catheter. AB - OBJECTIVES: A rational approach in the treatment of chronic rhinosinusitis (CRS) is the intranasal application of antiseptic agents, due to the pathogenetic role of bacteria and fungi. N-Chlorotaurine (NCT), a mild endogenous oxidant with broad-spectrum antimicrobial activity, has been tested for the first time in CRS. METHODS: This one-arm phase IIa clinical study is the first step in the clinical development of this promising substance for local therapy of CRS. The nasal and paranasal cavities of 12 patients were rinsed with 10-20 ml of 1% aqueous NCT solution, applied via a novel catheter system (YAMIK). Treatment consisted of three lavages per week for 4 weeks. RESULTS: NCT caused neither alterations of the mucosa nor burning pain during application. Nevertheless, the insertion of the catheter, the insufflation of the posterior cuff and the overpressure inside the sinuses after infiltration led to moderate pain in some patients. Mucosal swelling decreased in all subjects, nasal breathing could be improved in nine patients and impaired olfaction in seven. Polyps did not disappear within the 1 month period of the study. CONCLUSIONS: The good tolerability and possible beneficial effects of NCT encourage its further investigation in CRS. Despite some limitations the YAMIK catheter proved to be a convenient and safe device for rinsing the nasal and paranasal sinuses. PMID- 16040220 TI - Gadolinium dose optimisation in patients with multiple sclerosis: intra- and inter-individual comparisons. AB - OBJECTIVE: To assess the efficacy of single, double and triple doses of 1M gadolinium in the magnetic resonance (MR) evaluation of multiple sclerosis (MS) lesions in order to establish the best trade-off in terms of dose and number of injections. MATERIALS AND METHODS: Twenty-three patients with relapsing remitting or secondary progressive MS underwent MR scans of the brain, spine or both. Signal intensity, the number of enhancing lesions and the areas of MS plaque were measured in T1-weighted images before and after single, double or triple doses of contrast medium. Wilcoxon's two-sample test and the Mann-Whitney U-test were used to evaluate the differences between the groups, and between doses in the same patient. RESULTS: There was no significant difference in area signal intensity or the number of enhancing lesions between the group examined after a single injection of double-dose (DD) contrast medium and the group examined after multiple injections of a triple dose (TD). The difference in the areas of enhancement and signal intensity between a single dose (SD) and TD significantly increased after the second bolus. CONCLUSION: Gadolinium injections in MS patients can be optimised by giving a single DD bolus, which leads to the same results as a TD administered as two boli. PMID- 16040221 TI - Cyclic voiding cystouretrography without the use of fluoroscopic monitoring. AB - PURPOSE: To determine whether cyclic voiding cystourethrography (VCUG) examinations can be performed without using real-time fluoroscopic monitoring. MATERIALS AND METHODS: VCUG examinations were performed in 209 children (133 girls and 76 boys). In each child, it was performed in a cyclic manner (three consecutive cycles) without using fluoroscopic monitoring and one radiograph was taken in each cycle of the examination. All patients were sedated by midazolam prior to examination. The degree of vesicoureteric reflux (VUR) was graded for each of the kidney ureter units (KUU) (total 418 KUU) separately. Statistical analysis included the comparison of the presence and absence of VUR and three ordinal-matched comparisons of each cycle. RESULTS: VCUG was detected in 37.3% of the children (78/209) and 28.7% (120/418) of the KUU. The absence of or same degree of VUR in three consecutive cycles were obtained in 345 KUU and the agreement was calculated as 82.5%. The same degree and presence of reflux were found in 85 and 89 refluxing KUU, the same negative findings (absence of VUR) in 306 and 298 KUU, and discordant findings in 27 and 31 KUU (between the first and second and between first and third cycles), respectively. Therefore, the agreement was 93.5 and 92.5% between the first and second cycles (p=0.70) and between the first and the third cycles (p=0.15), respectively. CONCLUSION: Both the cyclic nature of this study and the results indicated that VCUG without real time fluoroscopic monitoring could be used where adequate fluoroscopic examination is not possible for children in whom VUR detection is necessary and impacts treatment. However, the responsibility of pediatric radiologist always must also include the task to provide proper equipment for imaging children with suspected VUR. PMID- 16040222 TI - Physico-chemical characterization of benzocaine-beta-cyclodextrin inclusion complexes. AB - Local anesthetics are able to induce pain relief by binding to the sodium channel of excitable membranes, blocking the influx of sodium ions and the propagation of the nervous impulse. Benzocaine (BZC) is a local anesthetic whose low water solubility limits its application to topical formulations. The present work focuses on the characterization of inclusion complexes of BZC in beta cyclodextrin (beta-CD). Differential scanning calorimetry and electron microscopy gave evidences of the formation and the morphology of the complex. Fluorescence spectroscopy showed a BZC/beta-CD 1:1 stoichiometry. Phase-solubility diagrams allowed the determination of the association constants between BZC and beta-CD (549 M(-1)) and revealed that a three-fold increase in BZC solubility can be reached upon complexation with beta-CD. The details of BZC/beta-CD molecular interaction were analyzed by 1H 2D NMR allowing the proposition of an inclusion model for BZC into beta-CD where the aromatic ring of the anesthetic is located near the head of the beta-CD cavity. Moreover, in preliminary toxicity studies, the complex seems to be less toxic than BZC alone, since it induced a decrease in the in vitro oxidation of human hemoglobin. These results suggest that the BZC/beta-CD complex represents an effective novel formulation to enhance BZC solubility in water, turning it promising for use outside its traditional application, i.e., in infiltrative anesthesia. PMID- 16040223 TI - Stability indicating LC method for the determination of pipamperone. AB - A high performance liquid chromatographic method for the determination of pipamperone in the presence of one related impurity and its degradation products is described. The method is based on the use of an amide functionalized bonded phase column (LC-ABZ+ Plus) and a mobile phase of acetonitrile-tetrahydrofuran sodium phosphate monobasic (0.05 M, pH 6.5) (16:11:73, v/v/v). All peaks are eluted in <8 min. The method was demonstrated to be precise, accurate and specific. Degradation study showed that the drug is stable in acidic medium while it degrades under basic and oxidative conditions. The results indicated that the proposed method could be used in a stability assay. PMID- 16040224 TI - LC determination of glimepiride and its related impurities. AB - Five impurities in glimepiride drug substance were detected and quantified using a simple isocratic reverse phase HPLC method. For the identification and characterization purpose these impurities were isolated from a crude reaction mixture of glimepiride using a normal phase HPLC system. Based on the spectroscopic data like NMR, FTIR, UV and MS these impurities were characterized and used as impurity standards for determining the relative response factor during the validation of the proposed isocratic reverse phase HPLC method. The chromatographic separation was achieved on a Phenomenex Luna C8 (2) 100 A, 5 microm, 250 mm x 4.6 mm using a mobile phase consisting of phosphate buffer (pH 7.0)-acetonitrile-tetrahydrofuran (73:18:09, v/v/v) with UV detection at 228 nm and a flow rate of 1 ml/min. The column temperature was maintained at 35 degrees C through out the analysis. The method has been validated as per international guidelines on method validation and can be used for the routine quality control analysis of glimepiride as active pharmaceutical ingredient (API). PMID- 16040225 TI - Prediction of gastro-intestinal absorption using multivariate adaptive regression splines. AB - Multivariate adaptive regression splines (MARS) and a derived method two-step MARS (TMARS) were used for modelling the gastro-intestinal absorption of 140 drug like molecules. The published absorption values for these molecules were used as response variable and calculated molecular descriptors as potential explanatory variables. Both methods were compared and their potential use in quantitative structure-activity relationship (QSAR) context evaluated. The predictive abilities of the models were studied using different sequences of Monte Carlo cross validation (MCCV). It was shown that both types of models had good predictive abilities and that for the data used, MARS gave better results than TMARS. It could be concluded that both methods could be valuable for QSAR modelling. PMID- 16040226 TI - Social status does not predict responses to Seoul virus infection or reproductive success among male Norway rats. AB - Trade-offs exist among life history strategies that are used to increase survival and reproduction; such that, males that engage in more competitive behaviors may be more susceptible to infection. Hantaviruses are transmitted horizontally between rodents through the passage of virus in saliva during wounding and male rodents are more likely to be infected with hantaviruses than females. To determine whether a trade-off exists between dominance and susceptibility to Seoul virus infection, male Long Evans rats were group housed (3/cage) with a female rat and aggressive and subordinate behaviors were monitored during a 10 day group housing condition. After behavioral testing, males were individually housed, inoculated with Seoul virus, and blood, saliva, and fecal samples were collected. Dominant males initiated more aggressive encounters than subordinate males. Dominant and subordinate males, however, had similar steroid hormone concentrations, anti-Seoul virus IgG responses, and weight gain over the course of infection. A similar proportion of dominant and subordinate males shed virus in saliva and feces during infection. Using microsatellite DNA markers paternity was assigned to pups derived during the group housing period. In contrast to our initial hypothesis, dominant and subordinate males sired a similar percentage of pups. Taken together, host social status may not predict reproductive success or susceptibility to hantaviruses in rodent reservoir populations. PMID- 16040227 TI - Aluminum-induced maternal and developmental toxicity and oxidative stress in rat brain: response to combined administration of Tiron and glutathione. AB - The current study was performed to assess the potential of 4,5-dihydroxy 1,3 benzene disulfonic acid di sodium salt (Tiron) and glutathione (GSH) either individually or in combination against aluminum (Al)-induced developmental toxicity in fetuses and sucklings of Wistar rats. Female rats were exposed to aluminum chloride at a dose of 345 mg/(kg day) oral from days 0 to 16 of gestation and 0 to 16 of post-partum (P.P.). Tiron and GSH were administered at a dose of 471 mg/(kg day) i.p. and 100 mg/(kg day) oral, respectively, on days 5, 7, 9, 11, 13, 15 and 17 of gestation and post-partum. Al caused reduction in number of corpora lutea, number of implantation sites, placental and fetal weight and stunted growth. Skeletal malformations were also observed in fetuses. Maternal toxicity was demonstrated by reduction in body weight gain. Induction of oxidative stress was also recorded in the brain of mother as well as in fetuses and sucklings after Al exposure. Significant decrease was recorded in reduced glutathione, glutathione reductase (GR), glutathione peroxidase (GPx), catalase (CAT), superoxide dismutase (SOD), acetyl cholinesterase (AChE) and increase was observed in TBARS and glutathione-S-transferase (GST) in brain of pregnant mothers, fetuses and sucklings. Most of the above parameters responded positively with individual therapy with Tiron, but more pronounced beneficial effects on the above-described parameters were observed when Tiron was administered in combination with GSH. Inductively coupled plasma-atomic emission spectroscopy (ICP-AES) studies also showed significantly high concentration of Al in suckling's brain and maternal blood, brain, placenta and fetal brain. Treatment with Tiron individually or in combination with glutathione, reduced the accumulation of the Al in almost all the organs studied. It is concluded that chelating agents reduced the Al-induced toxicity and Tiron was more effective in reducing blood Al concentration than glutathione when given individually. PMID- 16040228 TI - Monoisoamyl dimercaptosuccinic acid induced changes in pregnant female rats during late gestation and lactation. AB - Monoisoamyl dimercaptosuccinic acid (MiADMSA), a vicinal thiol chelating agent and an analogue of a conventional metal chelating agent, meso-2,3 dimercaptosuccinic acid (DMSA) has recently been gaining recognition to be more effective chelating agent than DMSA in mobilizing lead, mercury and arsenic. However, very little information is available on the toxicological properties of this chelator. In the present study, MiADMSA was administered to pregnant female rats from day 14 of gestation to day 21 of lactation at different doses through oral (p.o.) and intraperitoneal (i.p.) routes to examine the toxicity in the pups and dams. Results suggested that MiADMSA had no effect on period of gestation, litter-size, sex ratio, and viability and lactation. No skeletal defects were observed following the administration of the chelator. However, MiADMSA administration produced few signs of oxidative stress in dams particularly at the higher doses (100 and 200mg/kg) as evident from increased thiobarbituric acid reactive substances (TBARS) in RBCs and decrease in the delta-aminolevulinic acid dehydratase (ALAD) activity. Administration of MiADMSA also caused some alterations in the essential metal concentration in the soft tissues especially tissue copper loss in lactating mothers and pups, which would be of some concern. Apart from copper, changes were also observed in the tissue zinc concentrations in mothers and pups following MiADMSA administration. The study thus suggests that the chelator is relatively safe during late gestation and it does not cause any major alteration in the mothers and the developing pups. However, detailed studies with MiADMSA, post-toxic metal exposure in pregnant animals may provide useful information. PMID- 16040229 TI - Guidelines for the use of clozapine in individuals with developmental disabilities. AB - Clozapine is the most effective antipsychotic medication currently in use, but there has been a paucity of well-controlled research on its efficacy with people with developmental disabilities. We present a set of guidelines to ensure proper utilization of clozapine in individuals with developmental disabilities, because it can offer them therapeutic advantages similar to those observed in people with schizophrenia. We provide recommendations regarding the use of clozapine that are based on three main sources: literature and published professional practice guidelines regarding the use of clozapine in individuals who do not have developmental disabilities, the limited literature on the use of clozapine in individuals who have developmental disabilities, and our own clinical experience. The first part of the guidelines contains an overview of necessary practical knowledge regarding side effects, dose and blood level considerations, and interactions with other medications, diet and tobacco smoking. In the second part, we offer procedures for selecting individuals for clozapine therapy based on proper indications and contraindications for treatment. We also include requirements regarding informed consent, dosage and special laboratory and clinical monitoring. PMID- 16040230 TI - Prospects on behavioral studies of marine and freshwater toxins. AB - While there is a long-standing tradition of using behavioral methods to study the effects of manufactured drugs and environmental chemicals, comparatively little attention has focused until recently on the behavioral effects of marine or freshwater toxins. A vast array of microorganisms, found in a variety of waters, are known to occasionally "bloom" and produce toxins that can cause either blatant toxicity (i.e., lethality) or damage to a number of organ systems. The nervous system is a known target for many of the toxins. Considerable research has in the past been carried out to determine toxin effects on the survivability of laboratory rodents (typically mice) following acute exposures. Newer research has shown, however, prominent toxin-induced alterations in motor, sensory, autonomic and cognitive functions at sublethal exposure concentrations. Future toxin research can capitalize upon a wealth of behavioral paradigms already available in toxicology, pharmacology and neuroscience. PMID- 16040231 TI - The validity and utility of geotaxis in young rodents. AB - Negative geotaxis, an automatic, reliable, stimulus-bound, orientation and movement directionally against gravitational cues, is often used for behavioral assessments of infant rodents. We summarize historical and contemporary analyses and conclude that negative geotaxis does not exist in infant rats. Infant rodents placed on inclined surfaces (ranging from 15 degrees to 70 degrees in most tests) are posturally unstable and their compensatory responses have been misinterpreted as negative geotaxis. In fact, recent findings suggest that if infant rats display a geotaxis, they show positive geotaxis on shallow angles of inclination (e.g., 4 degrees and 8 degrees ). There may be utility in assessing postures and motoric responses of infant rats on relatively robust angles of inclination, but these are not tests of negative geotaxis. PMID- 16040233 TI - Long-term kindled seizures induce alterations in hematopoietic functions: role of serum leptin. AB - Recent studies conducted in our laboratory have demonstrated marked increases in both serum leptin levels and colony numbers in bone marrow progenitor cells following long-term kindled seizures in rats. The present study sought to determine whether such changes in hematopoietic functions following kindling are linked to increased serum leptin levels. Kindled stage V seizures were induced for 30 days in Sprague-Dawley rats by stimulation of the basal complex of amygdala. The results revealed colony numbers in colony forming units granulocyte/macrophage (CFU-GM) cultures from kindled rats increased significantly, an effect that was blocked by the presence of an anti-leptin antibody. The results further demonstrated that the addition of serum obtained from kindled rats to CFU-GM cultures from control rats significantly increased the numbers of colonies relative to non-serum added cultures. Moreover, the proliferative effects of serum from kindled rats were also blocked by adding an anti-leptin antibody. These findings were confirmed from the observations that the long isoform of the leptin receptor, which is capable of signal transduction, was present only in kindled, but not in control rats. Thus, the results provide evidence that the hematopoietic changes observed following long-term kindling are directly associated with elevated serum leptin levels. PMID- 16040234 TI - Phosphatidylinositol-dependent phospholipases C Plc2 and Plc3 of Candida albicans are dispensable for morphogenesis and host-pathogen interaction. AB - Phospholipases play an important role as virulence factors in human pathogens. Candida albicans, the major fungal pathogen of humans, encodes phospholipases of type A, B, C and D. Type B Plb2 and type D Pld1 phospholipases have been shown to contribute to virulence in this organism. We analyzed, in C. albicans, PLC2 and PLC3, two highly conserved genes coding for phosphatidylinositol-dependent phospholipases C with homology to the known virulence factor PlcA in the human pathogen Listeria monocytogenes. We show that expression of PLC2 and PLC3 is upregulated under different filament-inducing conditions and in the constitutive filamentous mutant tup1Delta. In order to analyze PLC2 and PLC3 function in C. albicans, we constructed strains that carry PLC2 or PLC3 under a constitutive promoter and strains that lack all four PLC2/3 alleles. These strains were not affected in their ability to produce filaments under non-inducing conditions, nor was filamentation modified under inducing conditions, suggesting that PLC2/3 are not critical determinants of the yeast-to-hypha switch. In a cell culture model for macrophage interaction, phagocytosis of C. albicans and subsequent killing were not influenced by PLC2/3. These results demonstrate that C. albicans PLC2 and PLC3 are dispensable for virulence; moreover, they underline the sharp contrast with the function of plcA in L. monocytogenes. PMID- 16040235 TI - Effect of outer membrane permeabilisation on intrinsic resistance to low triclosan levels in Pseudomonas aeruginosa. AB - The present study was undertaken to investigate the possibility that outer cell envelope impermeability might be involved in the intrinsic resistance of Pseudomonas aeruginosa to low levels of the hydrophobic biocide triclosan. Macrobroth dilution and batch cultural turbidimetric assays were employed to assess the ability of compounds that render the Gram-negative outer membrane permeable to non-polar molecules to sensitise cell envelope variants to triclosan. Pseudomonas aeruginosa strains possessing highly refractory (PAO1) and atypically permeable (Z61) outer cell envelopes as well as a PAO1 derivative lacking four multidrug efflux pumps (YM64) were examined. Whilst the triclosan minimal inhibitory concentrations (MICs) differed dramatically for both PAO1 and Z61, significant decreases were seen for both strains in the presence of the outer membrane permeabiliser polymyxin B-nonapeptide. Strain YM64 was as resistant to triclosan as strain PAO1. Turbidimetric assessments of batch cultural growth kinetics revealed that the three chemically unrelated outer membrane permeabilisers polymyxin B-nonapeptide, compound 48/80 and ethylenediaminetetraacetate (EDTA) sensitised all strains to a sub-MIC concentration of triclosan (2.0 microg/mL). These data support the notion that the outer membrane exclusionary properties of P. aeruginosa for non-polar molecules confer intrinsic resistance to low concentrations of triclosan such as might be expected to occur in environmental residues. Moreover, a role for outer cell envelope impermeability is suggested for resistance to high triclosan concentrations in vitro. PMID- 16040236 TI - Interstimulus contingency facilitates saccadic responses in a bimodal go/no-go task. AB - The saccadic response to a suddenly appearing visual target stimulus is faster when an accessory auditory stimulus is presented in its spatiotemporal proximity. This multisensory facilitation of reaction time is usually considered a mandatory bottom-up process. Here, we report that it can be modulated by the predictability of the target location provided by an accessory stimulus, thereby indicating a form of top-down processing. Subjects were asked to make a saccade in the direction of a visual target randomly appearing left or right from fixation. An accessory auditory stimulus was presented either at the same location or opposite to the target, with the probability varying over blocks of presentation. Thus, the auditory stimulus contained probabilistic information about the target location (interstimulus contingency). A certain percentage of the trials were catch trials in which the auditory accompanying stimulus (Experiment 1) or the visual target (Experiment 2) was presented alone and the subjects were asked to withhold their response. In particular with visual catch trials, varying the predictability of target location resulted in reaction time facilitation in the bimodal trials, with both high (80%) and low predictability (20%), but only when both stimuli were presented within a small time window (40 ms). As subjects could not possibly follow the task instructions in this short period explicitly, we conclude that they utilized the interstimulus contingency information implicitly, thus revealing an extremely fast involuntary top-down control on saccadic eye movements. PMID- 16040237 TI - Effects of silver nanoparticles on the fluidity of bilayer in phospholipid liposome. AB - This paper describes the formation and characterization of liposome entrapping the silver nanoparticles in bilayer. Silver nanoparticles were entrapped in the bilayer of dipalmitoylphosphatidylcholine (DPPC) liposome, named as silver-loaded liposome. Specifically, above the gel to liquid-crystalline phase transition temperature of this lipid (i.e., 41 degrees C), it was observed that membrane fluidities of silver-loaded liposomes were increased, and fluorescence anisotropy values were reduced from 0.114 to 0.097. This might be due to the structural modifications and interactions between DPPC molecules and silver nanoparticles within the bilayer. It was also confirmed that silver nanoparticles were entrapped in hydrophobic region of lipid bilayer with transmission electron microscopy (TEM) and electron energy loss spectroscopy (EELS) measurements. PMID- 16040238 TI - A novel FRET-based optical fiber biosensor for rapid detection of Salmonella typhimurium. AB - A biosensor that is portable and permits on-site analysis of samples would significantly reduce the large economical burden of food products recalls. A fiber optic portable biosensor utilizing the principle of fluorescence resonance energy transfer (FRET) was developed for fast detection of Salmonella typhimurium (S. typhimurium) in ground pork samples. Labeled antibody-protein G complexes were formed via the incubation of anti-Salmonella antibodies labeled with FRET donor fluorophores (Alexa Fluor 546) and protein G (PG) labeled with FRET acceptor fluorophores (Alexa Fluor 594). Utilizing silanization, the labeled antibodies-PG complexes were then immobilized on decladded, tapered silica fiber cores to form the evanescent wave-sensing region. The biosensors were tested in two different solutions: (1) PBS doped with S. typhimurium and (2) homogenized pork sample with S. typhimurium. The fiber probes tested in a S. typhimurium doped phosphate buffered solution demonstrated the feasibility of the biosensor for detecting S. typhimurium as well as determined the optimal packing density of the labeled antibody-PG complexes on the surface of fibers. The results showed that a packing density of 0.033 mg/ml produced the lowest limit of detection of 10(3)cells/ml with 8.2% change in fluorescence. The fiber probes placed in homogenized pork samples inoculated with S. typhimurium showed a limit of detection of 10(5)CFU/g with a 6.67% in fluorescence within a 5-min response time. These results showed that the FRET-based fiber optic biosensor can become a useful analytical tool for detection of S. typhimurium in real food samples. PMID- 16040239 TI - Analysis of anabolic steroids in the horse: development of a generic ELISA for the screening of 17alpha-alkyl anabolic steroid metabolites. AB - Due to the potential for misuse of a wide range of anabolic steroids in horse racing, a screening test to detect multiple compounds, via a common class of metabolites, would be a valuable forensic tool. An enzyme-linked immunosorbent assay (ELISA) has been developed to detect 17alpha-alkyl anabolic steroid metabolites in equine urine. 16beta-Hydroxymestanolone (16beta,17beta-dihydroxy 17alpha-methyl-5alpha-androstan-3-one) was synthesised in six steps from commercially available epiandrosterone (3beta-hydroxy-5alpha-androstan-17-one). Polyclonal antibodies were raised in sheep, employing mestanolone (17beta-hydroxy 17alpha-methyl-5alpha-androstan-3-one) or 16beta-hydroxymestanolone conjugated to human serum albumin, via a 3-carboxymethyloxime linker, as antigens. Antibody cross-reactivities were determined by assessing the ability of a library of 54 representative steroids to competitively bind the antibodies. Antibodies raised against 16beta-hydroxymestanolone showed excellent cross-reactivities for all of the 16beta,17beta-dihydroxy-17alpha-methyl steroids analysed and an ELISA has been developed to detect these steroid metabolites. Using this 16beta hydroxymestanolone assay, urine samples from horses administered with stanozolol (17alpha-methyl-pyrazolo[4',3':2,3]-5alpha-androstan-17beta-ol), were analysed raw, following beta-glucuronidase hydrolysis, and following solid-phase extraction (SPE) procedures. The suppressed absorbances observed were consistent with detection of the metabolite 16beta-hydroxystanozolol. Positive screening results were confirmed by comparison with standard LCMS analyses. Antibodies raised against mestanolone were also used to develop an ELISA and this was used to detect metabolites retaining the parent D-ring structure following methandriol (17alpha-methylandrost-5-ene-3beta,17beta-diol) administration. The ELISA methods developed have application as primary screening tools for detection of new and known anabolic steroid metabolites. PMID- 16040240 TI - Removal of reactive dyes from wastewater by adsorption on coir pith activated carbon. AB - The removal efficiency of activated carbon prepared from coir pith towards three highly used reactive dyes in textile industry was investigated. Batch experiments showed that the adsorption of dyes increased with an increase in contact time and carbon dose. Maximum de-colorisation of all the dyes was observed at acidic pH. Adsorption of dyes was found to follow the Freundlich model. Kinetic studies indicated that the adsorption followed first order and the values of the Lagergren rate constants of the dyes were in the range of 1.77 x 10(-2)-2.69 x 10(-2)min(-1). The column experiments using granular form of the carbon (obtained by agglomeration with polyvinyl acetate) showed that adsorption efficiency increased with an increase in bed depth and decrease of flow rate. The bed depth service time (BDST) analysis carried out for the dyes indicated a linear relationship between bed depth and service time. The exhausted carbon could be completely regenerated and put to repeated use by elution with 1.0M NaOH. The coir pith activated carbon was not only effective in removal of colour but also significantly reduced COD levels of the textile wastewater. PMID- 16040241 TI - Aqua mediated synthesis of substituted 2-amino-4H-chromenes and in vitro study as antibacterial agents. AB - A simple, clean, environmentally benign route to the synthesis of 2-amino chromenes is described using K2CO3 as a green catalyst in water under microwave irradiation. This implies a convenient route avoiding the usage of hazardous organic solvents and organic bases. This technique requires only water in both the reaction step and workup, thus rendering the whole procedure into a truly ecofriendly green protocol. All the synthesized compounds were shown to possess antibacterial activity as tested in vitro against standard strains of Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. PMID- 16040242 TI - A new synthesis of isoaurones: cytotoxic activity of compounds related to the alleged structure of isoaurostatin. AB - A new synthesis of isoaurones related to the alleged structure of isoaurostatin, via Heck intramolecular cyclization of cinnamic esters of 2-iodophenols, is reported. The cytotoxic activity of these isoaurones is lower than that of the structurally very similar 4-arylcoumarins. PMID- 16040243 TI - Two novel proteins, dos1 and dos2, interact with rik1 to regulate heterochromatic RNA interference and histone modification. AB - BACKGROUND: Chromosomal behavior during mitosis and meiosis depends in part on heterochromatic modifications such as histone H3 lysine-9 methylation (H3K9me). In fission yeast, the Heterochromatin Protein 1 homolog Swi6 recognizes H3K9me, silences transcription, and retains cohesin at pericentromeric repeats. Heterochromatin formation also depends on processing of transcripts derived from centromeric repeats by the RNAi machinery. The DDB1 homolog, Rik1, and histone methyltransferase, Clr4, act in a complex to promote H3K9me. However, the mechanism underlying this interaction is poorly understood. RESULTS: Using a cytological screen, we have identified two novel genes, dos1(+) and dos2(+), which are required for localization of Swi6. Deletion of either of these genes results in mitotic and meiotic chromosome missegregation, defects in mitotic centromeric cohesion and meiotic telomere clustering, and loss of heterochromatic silencing. Dos1 is predominantly located in the nucleus in a Dos2-dependent manner and directly interacts with Rik1. Each of these genes is required for the association of H3K9me with centromeric repeats, as well as for the production of small interfering RNAs. CONCLUSIONS: Dos1 and Dos2 are required for the formation of heterochromatin in fission yeast. We hypothesize that the physical interaction between Dos1 and Rik1 represents a role in regulating activity of the Rik1/Clr4 complex. Dos2 contributes to this role by regulating Dos1 localization. Our findings suggest a mechanism for recruitment of Clr4 in the RNAi-dependent heterochromatin pathway, in which Dos1 and Dos2 are essential. PMID- 16040244 TI - Partially redundant functions of Arabidopsis DICER-like enzymes and a role for DCL4 in producing trans-acting siRNAs. AB - Arabidopsis encodes four DICER-like (DCL) proteins. DCL1 produces miRNAs, DCL2 produces some virus-derived siRNAs, and DCL3 produces endogenous RDR2-dependent siRNAs, but the role of DCL4 is unknown. We show that DCL4 is the primary processor of endogenous RDR6-dependent trans-acting siRNAs (tasiRNAs). Molecular and phenotypic analyses of all dcl double mutants also revealed partially compensatory functions among DCL proteins. In the absence of DCL4, some RDR6 dependent siRNAs were produced by DCL2 and DCL3, and in the absence of DCL3, some RDR2-dependent siRNAs were produced by DCL2 and DCL4. Consistent with partial redundancies, dcl2 and dcl3 mutants developed normally, whereas dcl4 and dcl3 dcl4 mutants had weak and severe rdr6 phenotypes, respectively, and increased tasiRNA target mRNA accumulation. After three generations, dcl3 dcl4 and dcl2 dcl3 mutants exhibited stochastic developmental phenotypes, some of which were lethal, likely owing to the accumulated loss of heterochromatic siRNA-directed marks. dcl1 dcl3 and dcl1 dcl4, but not dcl1 dcl2 mutants, had phenotypes more severe than dcl1 mutants, consistent with DCL1, DCL3, and DCL4 acting as the primary processors of the three respective classes of endogenous silencing RNAs and DCL2 acting to produce viral-derived siRNAs and as an alternative DCL for endogenous siRNA production. PMID- 16040245 TI - Calcium elevation at fertilization coordinates phosphorylation of XErp1/Emi2 by Plx1 and CaMK II to release metaphase arrest by cytostatic factor. AB - BACKGROUND: Vertebrate oocytes are arrested at second meiotic metaphase by cytostatic factor (CSF) while awaiting fertilization. Accumulating evidence has suggested that inhibition of the anaphase-promoting complex/cyclosome (APC/C) is responsible for this arrest. Xenopus polo-like kinase 1 (Plx1) is required for activation of the APC/C at the metaphase-anaphase transition, and calcium elevation, upon fertilization/activation of eggs, acting through calmodulin dependent kinase II (CaMKII) is sufficient to activate the APC/C and terminate CSF arrest. However, connections between the Plx1 pathway and the CaMKII pathway have not been identified. RESULTS: Overexpression of Plx1 causes CSF release in the absence of calcium, and depletion of Plx1 from egg extracts blocks induction of CSF release by calcium and CaMKII. Prior phosphorylation of the APC/C inhibitor XErp1/Emi2 by CaMK II renders it a good substrate for Plx1, and phosphorylation by both kinases together promotes its degradation in egg extracts. The pathway is enhanced by the ability of Plx1 to cause calcium independent activation of CaMKII. The results identify the targets of CaMKII and Plx1 that promote egg activation and define the first known pathway of CSF release in which an APC/C inhibitor is targeted for degradation only when both CaMKII and Plx1 are active after calcium elevation at fertilization. CONCLUSIONS: Plx1 with an intact polo-box domain is necessary for release of CSF arrest and sufficient when overexpressed. It acts at the same level as CaMKII in the pathway of calcium-induced CSF release by cooperating with CaMKII to regulate APC/C regulator(s), such as XErp1/Emi2, rather than by directly activating the APC/C itself. PMID- 16040246 TI - BUR kinase selectively regulates H3 K4 trimethylation and H2B ubiquitylation through recruitment of the PAF elongation complex. AB - Histone-lysine methylation is linked to transcriptional regulation and the control of epigenetic inheritance. Lysine residues can be mono-, di-, or trimethylated, and it has been suggested that each methylation state of a given lysine may impart a unique biological function. In yeast, histone H3 lysine 4 (K4) is mono-, di-, and trimethylated by the Set1 histone methyltransferase. Previous studies show that Set1 associates with RNA polymerase II and demarcates transcriptionally active genes with K4 trimethylation. To determine whether K4 trimethylation might be selectively regulated, we screened a library of yeast deletion mutants associated with transcriptional regulation and chromatin function. We identified BUR2, a cyclin for the Bur1/2 (BUR) cyclin-dependent protein kinase, as a specific regulator of K4 trimethylation. Surprisingly, BUR also regulated H2B monoubiquitylation, whereas other K4 methylation states and H3 lysine 79 (K79) methylation were unaffected. Synthetic genetic array (SGA) and transcription microarray analyses of a BUR2 mutant revealed that BUR is functionally similar to the PAF, Rad6, and Set1 complexes. These data suggest that BUR acts upstream of these factors to control their function. In support, we show that recruitment of the PAF elongation complex to genes is significantly impaired in a BUR2 deletion. Our data reveal a novel function for the BUR kinase in transcriptional regulation through the selective control of histone modifications. PMID- 16040247 TI - Endothelial function markers in parkinsonian patients with hyperhomocysteinemia. AB - Hyperhomocysteinemia is considered a risk factor for vascular disease causing endothelial damage and consequently atherogenesis. The purpose of this study was to investigate the effect of elevated homocysteine on certain biochemical markers of endothelial function in patients with idiopathic Parkinson's disease (PD). Blood homocysteine levels were assessed in 57 PD patients and 40 matched normal controls. Investigation of the C677T 5,10 methylenetetrahydrofolate reductase (MTHFR) genotype was also performed in 43 PD patients. The following markers of endothelial function were assessed: superoxide dismutase (SOD), nitric oxide (NO), sICAM-1 and sE-selectin. Homocysteine levels were found mildly elevated in PD patients particularly in those treated with L-Dopa. MTHFR genotype did not influence significantly this finding. SOD activity was found reduced but it was not correlated to homocysteine levels. All other parameters measured were normal and were not related to hyperhomocysteinemia. Our findings indicate that mild hyperhomocysteinemia in PD patients was not associated with endothelial dysfunction. PMID- 16040248 TI - Transient partial ophthalmoplegia and Horner's syndrome after intraoral local anesthesia. AB - Local neurological symptoms and signs are infrequent after intraoral anesthesia for dental procedures, thus diagnosis may be challenging for a neurologist unfamiliar with this benign phenomenon. Unnecessary diagnostic procedures may be performed and can be associated with complications. We present a 19-year old woman with transient diplopia, miosis, partial enophthalmia and lacrimation on the side of injection after intraoral anesthesia with prilocaine. PMID- 16040249 TI - 'O-Acyl isopeptide method' for the efficient preparation of amyloid beta peptide 1-42 mutants. AB - Novel water-soluble isopeptides of Abeta1-42 mutants, '26-O-acyl isoAbeta1-42 (26 AIAbeta42) mutants', which were efficiently converted to intact Abeta1-42 mutants with no byproduct formation under physiological conditions, were synthesized. These isopeptides provide a new system useful for investigating the biological function of Abeta1-42 mutants. PMID- 16040250 TI - Effect of hypoxic preconditioning on brain genomic response before and following ischemia in the adult mouse: identification of potential neuroprotective candidates for stroke. AB - The aim of the present study is to better understand oxygen-sensitive adaptative pathways underlying the hypoxic preconditioning-induced protection of the brain against ischemia. Using oligonucleotide microarrays, we examined the brain genomic response of adult mice following hypoxia preconditioning (8% O2 for 1 or 6 h of hypoxia with reoxygenation 12, 18, 24 h or 72 h) and ischemia (6 h), preceeded (tolerant state) or not, by preconditioning. Real-time PCR was used to validate the results. Most gene expression increases occurred during hypoxia, including those of HIF-1-dependent genes (RTP801, AM, VEGF, p21, GLUT-1), early response genes (IER3) and transcriptional factors (ATF3, C/EBPdelta). A second wave of changes occurred 24 h after reoxygenation (S100A5, TH, Calretinin, PBX3). A third one occurred during ischemia itself, revealing that hypoxic preconditioning modifies the brain genomic response to ischemia. In addition, we show that some identical genes are overexpressed by hypoxia in both neonatal and adult brains (VEGF, EPO, GLUT-1, AM, MTs, C/EBPdelta). PMID- 16040251 TI - Monitoring photodynamic therapy of localized infections by bioluminescence imaging of genetically engineered bacteria. AB - The increasing occurrence of multi-antibiotic resistant microbes has led to the search for alternative methods of killing pathogens and treating infections. Photodynamic therapy (PDT) uses the combination of non-toxic dyes and harmless visible light to produce reactive oxygen species that can kill mammalian and microbial cells. Although the photodynamic inactivation of bacteria has been known for over a hundred years, its use to treat infections has not been much developed. This may be partly due to the difficulty of monitoring the effectiveness of PDT in animal models of infection. In order to facilitate this monitoring process, we have developed a procedure that uses bioluminescent genetically engineered bacteria and a light sensitive imaging system to allow real-time visualization of infections. When these bacteria are treated with PDT in vitro, the loss of luminescence parallels the loss of colony-forming ability. We have developed several models of infections in wounds and soft-tissue abscesses in mice that can be followed by bioluminescence imaging. The size and intensity of the infection can be sequentially monitored in a non-invasive fashion in individual mice in real-time. When photosensitizers are introduced into the infected tissue followed by illumination with red light, a light-dose dependent loss of luminescence is seen. If the bacterium is invasive, the loss of luminescence correlates with increased survival of the mice, whilst animals in control groups die of sepsis within five days. Healing of the PDT treated wounds is not impaired and may actually be improved. This approach can allow many animal models of localized infections to be accurately monitored for efficacy of treatment by PDT. PMID- 16040253 TI - B-lymphocyte-induced maturation protein-1 (Blimp-1) gene of torafugu (Takifugu rubripes). AB - Homologous gene of B-lymphocyte-induced maturation protein-1 (Blimp-1) of torafugu (Takifugu rubripes) was identified by tblast search analysis. RT-PCR and 5'RACE clearly defined the sequence of the UTR and coding region which has been ambiguously determined by tblast analysis. Fugu Blimp-1 was mainly expressed in the lymphoid organs. These finding imply that Blimp-1 would take a major role in the terminal differentiation of B-cells to plasma cells in fish. PMID- 16040252 TI - PEDF and the serpins: phylogeny, sequence conservation, and functional domains. AB - Pigment epithelium derived factor (PEDF) is non-inhibitory serpin with neurotrophic and antiangiogenic functions. In this study, we have assembled PEDF sequences for 9 additional species by data base mining and performed cross species alignment for 14 PEDF sequences to identify conserved structural domains. We found evolutionary conservation of a leader sequence, a single C-terminal glycosylation site, collagen-binding residues, and four specific conserved PEDF peptides. The C-terminus, 384--415 and an N-terminal region 78--95, show close homology with many other serpins, and there is strong conservation of 39 of 51 consensus key residues involved in serpin structure and function. Two peptide regions, 40--67 and 277--301, are unique to PEDF but conserved in all species. Conserved residues at the N-terminus, helix d (hD), and helix A (hA) of PEDF form a structure similar to the heparin-binding groove of other serpins. We identified a motif in PEDF that is homologous to the nuclear localization signals of other proteins. A bitopographical localization of PEDF was confirmed by immunocytochemistry and Western blots. Our results suggest that secretion is required for PEDF's activity, that PEDF can migrate to the nucleus, and that PEDF has structural and functional features more common with inhibitory serpins. PMID- 16040254 TI - Immunoglobulin protein and gene transcripts in sea bream (Sparus aurata L.) oocytes. AB - Cellular mechanisms of Ig transfer to Sparus aurata developing oocytes were analysed. Homologous serum Ig was purified and used to raise the rabbit polyclonal antiserum ORa. Immunohistochemistry revealed an active role of both follicular cells (already at a pre-vitellogenetic stage) and oocytes in the Ig uptake. Early vitellogenetic oocytes (lipid vesicle stages) had ORa staining of outer cortex and oolemma as well as of their follicular cells. In protein yolk granule oocytes, ORa staining was notably found in the pore canals crossing the egg envelope and at the periphery of yolk platelets. A transfer of Ig from the blood to the follicles appears likely. In addition, RT-PCR using specific primers for the constant region of sea bream Ig L chain detected Ig mRNA in released eggs and no signal in post-hatching larvae. These findings show that a significant level of Ig gene transcription in the oocyte and/or a transfer of transcripts may also occur. PMID- 16040255 TI - Crossing the blood-brain barrier: a potential application of myristoylated polyarginine for in vivo neuroimaging. AB - As basic neurological research continues to reveal novel targets for therapy, the need to deliver therapeutic agents across the blood-brain barrier (BBB) becomes increasingly important. If developed, delivery modules would bring targeting molecules across the BBB to their respective active sites. In addition, it would be highly advantageous if the bioavailability of these delivered agents could be monitored over time using non-invasive imaging techniques. Here, we describe a versatile delivery module based on a myristoylated polyarginine backbone, which crosses the BBB. Incorporation of the fatty acid group was achieved using a Schotten-Bauman reaction with quantitative yield, and the peptide was further synthesized by conventional solid phase peptide synthesis (SPPS). We report for the first time the in vivo distribution of the delivery module over time into mouse brain using near-infrared (NIR) fluorescence imaging. The fluorescent cargo was detected in vivo from 24-48 h post IV injection and was further characterized in perfused brains. Immunohistochemical staining of excised brain showed that the delivery module primarily accumulated in neurons with occasional localization in astrocytes and endothelial cells. We conclude that this approach can be used for the delivery of imaging probes and potentially targeted therapeutics across the BBB. PMID- 16040256 TI - Whole-head MEG analysis of cortical spatial organization from unilateral stimulation of median nerve in both hands: no complete hemispheric homology. AB - We examined the contralateral hemispheric cortical activity in MEG (151 ch) after unilateral median nerve stimulation of the right and left hand in twenty healthy right-handed subjects. The goal was to establish parameters to describe cortical activity of the hemispheric responses and to study the potential ability to assess differences in volunteers and patients. We focused on the within-subject similarity and differences between evoked fields in both hands. Cortical activity was characterized by the overlay display of waveforms (CWP), number of peak stages, loci of focal maxima and minima in each stage, 3D topographic maps and exemplified equivalent current dipole characteristics. The paired-wise test was used to analyze the hemispheric differences. The waveform morphology was unique across the subjects, similar CWPs were noted in both hemispheres of the individual. The contralateral hemispheric responses showed a well defined temporal-spatial activation of six to seven stages in the 500 ms window. Consistently (in over 80% of subjects), the six stages across the subjects were 20M, 30M, 50M, 70M, 90M, and 150M. A 240M was present in the left hemisphere (LH) in 15/20 subjects and in the right hemisphere (RH) in 10/20. Statistics of the latencies and amplitudes of these seven stages were calculated. Our results indicated that the latency was highly consistent and exhibited no statistical mean difference for all stages. Furthermore, no mean amplitude differences between both hemispheres at each stage were found. The patterns of magnetic fields in both hemispheres were consistent in 70% of the subjects. A laterality index (L.I.) was used for defining the magnetic field amplitude differences between two hemispheres for each individual. Overall, the absolute amplitude of the brain responses was larger in the left than in the right hemisphere in the majority of subjects (16/20), yet a significant portion (4/20) exhibited right dominance of the N20m activity. Each individual exhibited a unique CWP, there was reliable consistency of peak latencies and mean amplitudes in median nerve MEG. Nevertheless, this study indicates the limitations of using the intact hemisphere responses to compare with those from the affected (brain) side and suggests caution in assuming full homology in the cortical organization of both hemispheres. This study provides some results to address clinical issues like which parameter describes individual differences best. Whether a genuine difference is found or whether any difference may simply represent the variability encountered in a normal population. PMID- 16040257 TI - COMT Val108/158Met genotype affects the mu-opioid receptor system in the human brain: evidence from ligand-binding, G-protein activation and preproenkephalin mRNA expression. AB - Recent data from [(11)C]carfentanil ligand-PET indicate that in the human brain, the availability of mu-opioid (MOP) receptor binding sites is affected by the Val(108/158)Met polymorphism of the catechol-O-methyltransferase (COMT) gene. This prompted us to validate the impact of COMT Val(108/158)Met on MOP receptors in human post-mortem brain. [(3)H]DAMGO receptor autoradiography was performed in frontal cortex, basal ganglia, thalamus and cerebellum (8 Met/Met, 6 Met/Val, 3 Val/Val). With respect to genotype, numbers of MOP binding sites in COMT Met(108/158) homozygous and Val(108/158)Met heterozygous cases were higher than in Val(108/158) homozygous. Differences were significant in the caudate nucleus (Val/Met vs. Val/Val), nucleus accumbens (Val/Met vs. Val/Val) and the mediodorsal nucleus of the thalamus (Met/Met vs. Val/Val). In the thalamus, this was corroborated by DAMGO-stimulated [(35)S]GTPgammaS autoradiography. Moreover, stepwise multiple regression taking into account various covariables allowed to confirm the COMT genotype as the most predictive factor in this structure. As a mechanism how COMT might exert its action on MOP receptors, it has been suggested that at least in striatopallidal circuits COMT Val(108/158)Met impacts on enkephalin, which is capable of reciprocally regulating MOP receptor expression. Thus, we assessed preproenkephalin mRNA by in situ hybridization. In the striatum, mRNA levels were significantly higher in COMT Met(108/158) homozygous cases indicating that MOP binding sites and enkephalin are regulated in parallel. Moreover, the transcript was not detectable in the thalamus. Thus, mechanisms other than an enkephalin-dependent receptor turnover must be responsible for COMT related differences in MOP binding site availability in the human brain. PMID- 16040258 TI - Characterization of compound mechanisms and secondary activities by BioMAP analysis. AB - INTRODUCTION: Unexpected drug activities account for many of the failures of new chemical entities in clinical trials. These activities can be target-dependent, resulting from feedback mechanisms downstream of the primary target, or they can occur as a result of unanticipated secondary target(s). Methods that would provide rapid and efficient characterization of compounds with respect to a broad range of biological pathways and mechanisms relevant to human disease have the potential to improve preclinical and clinical success rates. METHODS: BioMAP assays containing primary human cells (endothelial cells and co-cultures with peripheral blood leukocytes) were stimulated in complex formats (specific combinations of inflammatory mediators) for 24 h in the presence or absence of test agents (drugs, experimental compounds, etc.). The levels of selected protein readouts (adhesion receptors, cytokines, enzymes, etc.) were measured and activity profiles (normalized data sets comprising BioMAP profiles) were generated for each test agent. The resulting profiles were compared by statistical methods to identify similarities and mechanistic insights. RESULTS: Compounds with known mechanisms including inhibitors of histamine H1 receptor, angiotensin converting enzyme, IkappaB kinase-2, beta2 adrenergic receptor and others were shown to generate reproducible and distinguishable BioMAP activity profiles. Similarities were observed between compounds targeting components within the same signal transduction pathway (e.g. NFkappaB), and also between compounds that share secondary targets (e.g. ibuprofen and FMOC-L-leucine, a PPARgamma agonist). DISCUSSION: Complex primary cell-based assays can be applied for detecting and distinguishing unexpected activities that may be of relevance to drug action in vivo. The ability to rapidly test compounds prior to animal or clinical studies may reduce the number of compounds that unexpectedly fail in preclinical or clinical studies. PMID- 16040259 TI - HLA-G in murine peripheral blood after interruption of pregnancy. AB - HLA-G antigens are highly expressed in maternal peripheral blood during early pregnancy in transgenic mice. In this study, we determined the levels of HLA-G in white blood cells during early pregnancy and after interruption of pregnancy in triple transgenic mice (H-2K(b)/HLA-G, hbeta2m, and hCD2/hCD8-TRI). The pregnancies were interrupted on day 7 using the anti-progesterone agent mifepristone (RU486). Blood samples of 20 pregnant TRI mice were taken and the HLA-G levels were determined on days 2, 4 and 6 of pregnancy and on days 9, 11 and 13 after fertilization. The monoclonal antibody W6/32, specific for monomorphic determinant HLA class I molecules, was used in combination with an immunolocalization method using a photonic microscope. The HLA-G levels increased gradually on days 2, 4 and 6 of pregnancy, and the interruption of pregnancy on day 7 was followed by a decrease of HLA-G levels. The data indicate that pregnancy is characterized by the early presence of HLA-G in maternal peripheral blood in TRI transgenic mice and suggest that HLA-G may serve as a useful indicator for pregnancy maintenance and well-being. PMID- 16040260 TI - Ileal impaction in 22 cows. AB - The clinical signs, diagnosis, treatment and outcome of 22 cows with ileal impaction were investigated using the medical records of bovine patients referred to the Department of Farm Animals, Vetsuisse Faculty, University of Zurich from 1993 to 2003. Only 15 of the cows had signs of colic, which were subtle but slowly increased in severity in some patients. The results of haematological and biochemical analyses were mildly abnormal in only few animals. There was no correlation between the duration of the disorder before admission, the severity of symptoms and the results of the haematological and biochemical analyses. Dilated loops of small intestine in the right dorsal quadrant of the abdomen could be palpated transrectally and imaged via ultrasonography. A definitive diagnosis of ileal impaction was made during exploratory laparotomy by finding the impaction and ruling out other abnormalities. In 19 cows, the obstructing food mass was easily massaged into the caecum, and in three animals an enterotomy was carried out. All cows had an uneventful recovery with no recurrence of the disorder. It is concluded that the cause of the impaction was most likely due to seasonal influences and winter-feeding with a hay based ration. The short and long-term prognosis after surgical intervention was good. PMID- 16040261 TI - Concentration by centrifugation for gas exchange EPR oximetry measurements with loop-gap resonators. AB - Measurement of the bimolecular collision rate between a spin label and oxygen is conveniently carried out using a gas permeable plastic sample tube of small diameter that fits a loop-gap resonator. It is often desirable to concentrate the sample by centrifugation in order to improve the signal-to-noise ratio (SNR), but the deformable nature of small plastic sample tubes presents technical problems. Solutions to these problems are described. Two geometries were considered: (i) a methylpentene polymer, TPX, from Mitsui Chemicals, at X-band and (ii) Teflon tubing with 0.075 mm wall thickness at Q-band. Sample holders were fabricated from Delrin that fit the Eppendorf microcentrifuge tubes and support the sample capillaries. For TPX, pressure of the sealant at the end of the sample tube against the Delrin sample holder provided an adequate seal. For Teflon, the holder permitted introduction of water around the tube in order to equalize pressures across the sealant during centrifugation. Typically, the SNR was improved by a factor of five to eight. Oxygen accessibility applications in site directed spin labeling studies are discussed. PMID- 16040262 TI - HoxA10-like proteins in the reproductive tract of the viviparous lizard Eulamprus tympanum and the oviparous lizard Lampropholis guichenoti. AB - The gene HoxA10 and its protein product are essential for the formation of the extensions of the plasma membrane called uterodomes or pinopods in mammalian uterine epithelia. In mice, the presence of the HoxA10 protein and uterodomes is needed for uterine receptivity to blastocyst implantation. The viviparous lizard Eulamprus tympanum displays uterodomes whereas the oviparous lizard Lampropholis guichenoti does not. To explore the theory that HoxA10 is involved in the formation of uterodomes we investigated whether HoxA10 immunoreactive proteins were present in both species during their reproductive cycles. Oviduct proteins from vitellogenic, gravid or non-reproductive L. guichenoti (n=19) and E. tympanum (n=28) were separated by electrophoresis and analysed by Western blot and specific antibodies to HoxA10. E. tympanum displayed HoxA10 immunoreactive bands at 59 and 63 kDa in 20 out of the 28 samples. All of the L. guichenoti samples displayed HoxA10 immunoreactive bands, 18 had bands at 59 and 64 kDa and 1 animal had a single band at 59 kDa. There were no significant differences in the level of HoxA10 immunoreactivity between the different stages of reproductive cycle in either species. The different molecular mass of the larger band in L. guichenoti (64 kDa) compared to E. tympanum (63 kDa) indicates that the two lizards express different isoforms of the HoxA10-like proteins and it will be interesting in future studies to determine whether there are differences in the biological activity of the proteins that regulate different physiological functions in the uterus of viviparous and oviparous lizards. PMID- 16040263 TI - Myoadenylate deaminase deficiency caused by alternative splicing due to a novel intronic mutation in the AMPD1 gene. AB - We have examined two Caucasian brothers with myoadenylate deaminase (AMPD) deficiency who presented with exercise intolerance and muscle cramps. Allele specific PCR amplification assays demonstrated that the common Q12X (C34T) and P48L (C143T) mutations were not found within their AMPD1 genes. Further analysis revealed that both brothers were compound heterozygotes for a previously reported K287I (A860T) mutation in exon 7 and a novel deletion within intron 2 (IVS2-(4 7)delCTTT). The intronic deletion appears to affect the splicing machinery since characterization of AMPD1 mRNA from skeletal muscle of one brother identified multiple alternatively spliced transcripts resulting in multiple deletions in exon 3, the complete deletion of either exon 3 or exons 3 and 4, and the activation of a cryptic splice site that resulted in an insertion at the 5' end of exon 4. The predominant transcript contains a 51 base deletion at the 5' end of exon 3 that is predicted to produce a functional form of AMPD containing a 17 amino acid residue deletion within its N-terminal region. Analysis of 137 Caucasian normal control patients determined that the K287I mutation is relatively frequent (5.1% carrier frequency), whereas the IVS2-(4-7)delCTTT mutation is rare and not present in 274 chromosomes. PMID- 16040265 TI - Clinical and nutritional evaluation of phenylketonuric patients on tetrahydrobiopterin monotherapy. AB - The clinical, nutritional, and neuropsychological data of 11 mild/moderate PKU patients after one year of treatment with BH4 are evaluated. BH4 monotherapy was introduced at 5 mg/kg/day in 14 PKU patients. In 11/14 patients, Phe tolerance increased significantly from 356+/-172 to 1546+/-192 mg/day (p=0.004), and special PKU formula was gradually reduced until complete removal. In them, mean plasma Phe concentrations remained below 360 micromol/L at 5 mg BH4/kg/day (7 mg/kg/day in one patient). BH4 therapy was stopped in three patients (V388M/P362T and R243Q/IVS10-11G>A genotypes) because it was not possible to improve Phe tolerance and to remove formula intake. Serum micronutrients were not significantly different at the start of treatment and at one year follow-up, except for selenium, which increased significantly after one year of therapy (p=0.017). Anthropometric, and nutritional measurements were within the age- and sex-specific percentiles for a healthy population after one year therapy. Neuropsychological follow-up indicated that intelligence scores persisted within normal limits. In terms of patients' genotype, we confirmed that the P275S mutation combined with R408W was associated with long-term BH4 responsiveness, while the combination of P362T/V388M, and R243Q/IVS10-11G>A resulted in poor metabolic control in long-term BH4 therapy. In summary, our data confirm that BH4 is a safe, and effective therapy in a selected group of mild, and moderate PKU patients who respond to the BH4 loading test. Low doses of BH4 in monotherapy permit withdrawal of the special formula and guarantee a good clinical and nutritional outcome with no adverse side effects in PKU patients. PMID- 16040264 TI - Effect of optimal dietary therapy upon visual function in children with long chain 3-hydroxyacyl CoA dehydrogenase and trifunctional protein deficiency. AB - The objective of this prospective cohort study was to determine if dietary therapy including docosahexaenoic acid (DHA; C22:6omega-3) supplementation prevents the progression of the severe chorioretinopathy that develops in children with long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) or trifunctional protein (TFP) deficiency. Physical, biochemical, and ophthalmological evaluations, including electroretinogram (ERG) and visual acuity by evoked potential (VEP), were performed at baseline and annually following the initiation of 65-130 mg/day DHA supplementation and continued treatment with a low-fat diet. Fourteen children with LCHAD or TFP deficiency, 1-12 years of age at enrollment, were followed for 2-5 years. Three subjects with TFP beta-subunit mutations had normal appearance of the posterior pole of the ocular fundi at enrollment and no changes over the course of the study. Eleven subjects who were homozygote and heterozygote for the common mutation, c.1528G>C, had no change to severe progression of atrophy of the choroid and retina with time. Of these, four subjects had marked to severe chorioretinopathy associated with high levels of plasma hydroxyacylcarnitines and decreased color, night and/or central vision during the study. The plasma level of long-chain 3-hydroxyacylcarnitines, metabolites that accumulate as a result of LCHAD and TFP deficiency, was found to be negatively correlated with maximum ERG amplitude (Rmax) (p=0.0038, R2=0.62). In addition, subjects with sustained low plasma long-chain 3 hydroxyacylcarnitines maintained higher ERG amplitudes with time compared to subjects with chronically high 3-hydroxyacylcarnitines. Visual acuity, as determined with the VEP, appeared to increase with time on DHA supplementation (p=0.051) and there was a trend for a positive correlation with plasma DHA concentrations (p=0.075, R2=0.31). Thus, optimal dietary therapy as indicated by low plasma 3-hydroxyacylcarnitine and high plasma DHA concentrations was associated with retention of retinal function and visual acuity in children with LCHAD or TFP deficiency. PMID- 16040266 TI - A simulation tool to support teaching and learning the operation of X-ray imaging systems. AB - We present a software tool for the simulation of an X-ray imaging systems. It consists of three virtual objects: the X-ray source, the human body and the detector. The X-ray source is modeled as a radiological tube for which the user can modify the tube potential, the anode material, the tube load, the filtration and some geometric parameters, such as source-skin distance, orientation and field size. The virtual body consists of a 3D voxel matrix in which CT numbers for each point of the body are stored, obtained from tomographic slices. The interactions of X-rays passing through the body are evaluated using pencil beam technique. The image is obtained computing the dose absorbed by the detector and converting it into optical density by the use of a proper response function. The dose absorbed in each point of the body is also computed and can be visualized both in 2D and 3D representations. The influence of each parameter on the beam spectrum, on the image quality and on the dose to the patient can be observed interactively. PMID- 16040267 TI - A case study of successful e-learning: a web-based distance course in medical physics held for school teachers of the upper secondary level. AB - Learning activities and course design in the new context of e-learning, such as in web-based courses involves a change both for teachers and students. The paper discusses factors important for e-learning to be successful. The development of an online course in medical physics and technology for high school teachers of physics, details of the course, and experience gained in connection with it are described. The course syllabus includes basics of radiation physics, imaging techniques using ionizing or non-ionizing radiation, and external and internal radiation therapy. The course has a highly didactic approach. The final task is for participants to design a course of their own centered on some topic of medical physics on the basis of the knowledge they have acquired. The aim of the course is to help the teachers integrate medical physics into their own teaching. This is seen as enhancing the interest of high school students in later studying physics, medical physics or some other branch of science at the university level, and as increasing the knowledge that they and people generally have of science. It is suggested that the basic approach taken can also have applicability to the training of medical, nursing or engineering students, and be used for continuing professional development in various areas. PMID- 16040268 TI - White matter tractography by anisotropic wavefront evolution and diffusion tensor imaging. AB - Determination of axonal pathways provides an invaluable means to study the connectivity of the human brain and its functional network. Diffusion tensor imaging (DTI) is unique in its ability to capture the restricted diffusion of water molecules which can be used to infer the directionality of tissue components. In this paper, we introduce a white matter tractography method based on anisotropic wavefront propagation in diffusion tensor images. A front propagates in the white matter with a speed profile governed by the isocontour of the diffusion tensor ellipsoid. By using the ellipsoid, we avoid possible misclassification of the principal eigenvector in oblate regions. The wavefront evolution is described by an anisotropic version of the static Hamilton-Jacobi equation, which is solved by a sweeping method in order to obtain correct arrival times. Pathways of connection are determined by tracing minimum-cost trajectories using the characteristic vector field of the resulting partial differential equation. A validity index is described to rate the goodness of the resulting pathways with respect to the directionality of the tensor field. Connectivity results using normal human DTI brain images are illustrated and discussed. We also compared our method with a similar level set-based tractography technique, and found that the anisotropic evolution increased the validity index of the obtained pathways by 18%. PMID- 16040269 TI - Reaching out of the shade. AB - Competition for light determines the success of individual plants in dense vegetation. Much depends on the capacity of plants to detect neighbours quickly and on their ability to respond to these signals. Recent findings indicate that although red:far-red ratios, and thus phytochromes, are of major importance in shade-avoidance responses, they do not act alone. Differences in light intensity also provoke shade-avoidance phenotypes, with blue light playing an important role in dense stands. Moreover, links between shade-avoidance signalling and auxins, gibberellins and ethylene have emerged. Additional breakthroughs are based on transcriptome studies that have unveiled new components in the response to shading. Amongst these, the phytochrome interacting factor 3-like proteins PIL1 and PIL2 underline the importance of circadian gating in shade avoidance. PMID- 16040270 TI - DNA metabolism and genetic diversity in Trypanosomes. AB - Trypanosomes are protozoan parasites that cause major diseases in humans and other animals. Trypanosoma brucei and Trypanosoma cruzi are the etiologic agents of African and American Trypanosomiasis, respectively. In spite of large amounts of information regarding various aspects of their biology, including the essentially complete sequences of their genomes, studies directed towards an understanding of mechanisms related to DNA metabolism have been very limited. Recent reports, however, describing genes involved with DNA recombination and repair in T. brucei and T. cruzi, indicated the importance of these processes in the generation of genetic variability, which is crucial to the success of these parasites. Here, we review these data and discuss how the DNA repair and recombination machineries may contribute to strikingly different strategies evolved by the two Trypanosomes to create genetic variability that is needed for survival in their hosts. In T. brucei, two genetic components are critical to the success of antigenic variation, a strategy that allows the parasite to evade the host immune system by periodically changing the expression of a group of variant surface glycoproteins (VSGs). One component is a mechanism that provides for the exclusive expression of a single VSG at any one time, and the second is a large repository of antigenically distinct VSGs. Work from various groups showing the importance of recombination reactions in T. brucei, primarily to move a silent VSG into an active VSG expression site, is discussed. T. cruzi does not use the strategy of antigenic variation for host immune evasion but counts on the extreme heterogeneity of their population for parasite adaptation to different hosts. We discuss recent evidence indicating the existence of major differences in the levels of genomic heterogeneity among T. cruzi strains, and suggest that metabolic changes in the mismatch repair pathway could be an important source of antigenic diversity found within the T. cruzi population. PMID- 16040271 TI - Analysis of the contribution of the beta-oxidation auxiliary enzymes in the degradation of the dietary conjugated linoleic acid 9-cis-11-trans octadecadienoic acid in the peroxisomes of Saccharomyces cerevisiae. AB - Beta-oxidation of the conjugated linoleic acid 9-cis,11-trans-octadecadienoic acid (rumenic acid) was analyzed in vivo in Saccharomyces cerevisiae by monitoring polyhydroxyalkanoate production in the peroxisome. Polyhydroxyalkanoate is synthesized by the polymerization of the beta-oxidation intermediates 3-hydroxyacyl-CoAs via a bacterial polyhydroxyalkanoate synthase targeted to the peroxisome. The amount of polyhydroxyalkanaote synthesized from the degradation of rumenic acid was found to be similar to the amount synthesized from the degradation of 10-trans,12-cis-octadecadienoic acid, oleic acid or 10 cis-heptadecenoic acid. Furthermore, the degradation of 10-cis-heptadecenoic acid was found to be unaffected by the presence of rumenic acid in the media. Efficient degradation of rumenic acid was found to be independent of the Delta(3,5),Delta(2,4)-dienoyl-CoA isomerase but instead relied on the presence of Delta(3),Delta(2)-enoyl-CoA isomerase activity. The presence of the unsaturated monomer 3-hydroxydodecenoic acid in polyhydroxyalkanoate derived from rumenic acid degradation was found to be dependent on the presence of a Delta(3),Delta(2) enoyl-CoA isomerase activity. Together, these data indicate that rumenic acid is mainly degraded in vivo in S. cerevisiae through a pathway requiring only the participation of the auxiliary enzymes Delta(3),Delta(2)-enoyl-CoA isomerase, along with the enzyme of the core beta-oxidation cycle. PMID- 16040272 TI - Reorganization of sleep patterns in severe OSAS under prolonged CPAP treatment. AB - OBJECTIVE: To evaluate the immediate and long-term recovery processes of sleep and daytime vigilance in patients with sleep apnea syndrome (OSAS) after continuous CPAP treatment. METHODS: Five consecutive polysomnographic (PSG) studies were carried out on 10 male patients with severe OSAS. The first recording (baseline) was accomplished without ventilatory support (N0). The other 4 recordings were carried out during the CPAP titration night (N1), during the second night of treatment (N2), during the third night of treatment (N3), and after 30 days of regular CPAP use (N30). Ten age-balanced healthy male subjects were selected from the Parma Sleep Center database as controls. Respiratory variables, conventional PSG variables, arousals, CAP (cyclic alternating pattern) variables, and daytime function (including MSLT) were quantified. ANOVA followed by post-hoc tests explored the differences between controls and OSAS patients in the different recording conditions (N0, N1, N2, N3, N30). The PSG measures that showed significant ANOVA values were correlated with the MSLT scores. RESULTS: Values of control subjects were recovered by REM sleep, REM latency, subtypes A3 and arousal index during N1, by CAP rate and total arousals during N2, by deep sleep (stages 3 + 4) during N3, by light sleep (stages 1 + 2) during N30. The only measures which remained below control values even after 1 month of sustained treatment were the amount of CAP cycles and A1 subtypes. MSLT scores correlated significantly with CAP rate, deep sleep and arousals. CONCLUSIONS: The changes induced by CPAP treatment do not restore immediately a normal sleep structure, which is re-established with different time scales SIGNIFICANCE: The modifications of sleep patterns and the different adjustments of phase A subtypes allow us to monitor the reorganization of sleep in OSAS patients treated with CPAP and the hierarchy of the mechanisms involved in sleep regulation. PMID- 16040273 TI - The diagnosis of heart failure in European primary care: The IMPROVEMENT Programme survey of perception and practice. AB - OBJECTIVE: To examine European primary care physicians (PCPs) views on diagnosis of heart failure and compare perceptions with actual practice. DESIGN: Semi structured PCP interviews and case note review on a random sample of heart failure patients. PARTICIPANTS: 1363 primary care physicians from 14 countries and 11,062 patient notes. MAIN OUTCOME MEASURES: Perceptions of PCPs compared to actual performance in heart failure (HF) diagnosis. RESULTS: Over 50% of patients with HF were above 70 years of age. Most subjects presented with typical clinical symptoms and objective signs of HF. In 50% of cases, HF was mainly diagnosed by PCPs. New York Heart Association classification was used by 50% of physicians. Electrocardiogram and chest X-ray were the most used diagnostic tests (90% and 84% respectively). PCPs considered echocardiography as having low diagnostic value, with only 48% routine usage. However, in actual practice echocardiography was used in 82% of diagnoses. Systolic dysfunction was observed in 51% HF subjects, but only 50% of physicians would differentiate systolic from diastolic heart failure. CONCLUSIONS: There was low use of NYHA classification (which denotes symptom severity) and differentiation between systolic and diastolic causes (which determines treatment strategies). PMID- 16040276 TI - Pattern-constrained multiple polypeptide sequence alignment. AB - Multiple sequence alignment (MSA) is one of the fundamental research topics in computational biology. The alignments help us to find functional assignment, evolutionary history and conserved region. Previous methods use a substitution matrix and do not incorporate knowledge of the sequences being aligned. Therefore, they do not assure the alignment of similar structures and common patterns in the sequences. We have been investigating into the solution to the problem in multiple and making use of knowledge of the sequences being aligned, including patterns in the Prosite databank, Blocks+, eBlocks databases, as well as motif and structural information. A pattern-constrained algorithm has been developed. Experiments with protein sequences have shown more accurate alignments with incorporation of the domain knowledge available in the sequences. PMID- 16040277 TI - Reconstruction of large phylogenetic trees: a parallel approach. AB - Reconstruction of phylogenetic trees for very large datasets is a known example of a computationally hard problem. In this paper, we present a parallel computing model for the widely used Multiple Instruction Multiple Data (MIMD) architecture. Following the idea of divide-and-conquer, our model adapts the recursive-DCM3 decomposition method [Roshan, U., Moret, B.M.E., Williams, T.L., Warnow, T, 2004a. Performance of suptertree methods on various dataset decompositions. In: Binida-Emonds, O.R.P. (Eds.), Phylogenetic Supertrees: Combining Information to Reveal the Tree of Life, vol. 3 of Computational Biology, Kluwer Academics, pp. 301-328; Roshan, U., Moret, B.M.E., Williams, T.L., Warnow, T., 2004b. Rec-I DCM3: A Fast Algorithmic Technique for reconstructing large phylogenetic trees, Proceedings of the IEEE Computational Systems Bioinformatics Conference (ICSB)] to divide datasets into smaller subproblems. It distributes computation load over multiple processors so that each processor constructs subtrees on each subproblem within a batch in parallel. It finally collects the resulting trees and merges them into a supertree. The proposed model is flexible as far as methods for dividing and merging datasets are concerned. We show that our method greatly reduces the computational time of the sequential version of the program. As a case study, our parallel approach only takes 22.1h on four processors to outperform the best score to date (Found at 123.7h by the Rec-I-DCM3 program [Roshan, U., Moret, B.M.E., Williams, T.L., Warnow, T, 2004a. Performance of suptertree methods on various dataset decompositions. In: Binida-Emonds, O.R.P. (Eds.), Phylogenetic Supertrees: Combining Information to Reveal the Tree of Life, vol. 3 of Computational Biology, Kluwer Academics, pp. 301-328; Roshan, U., Moret, B.M.E., Williams, T.L., Warnow, T., 2004b. Rec-I-DCM3: A Fast Algorithmic Technique for reconstructing large phylogenetic trees, Proceedings of the IEEE Computational Systems Bioinformatics Conference (ICSB)] on one dataset. Developed with the standard message-passing library, MPI, the program can be recompiled and run on any MIMD systems. PMID- 16040278 TI - Specific regression of human cancer cells by ribozyme-mediated targeted replacement of tumor-specific transcript. AB - In this study, we describe a novel approach to human cancer therapy that is based upon trans-splicing ribozyme-mediated replacement of cancer-specific RNAs with new transcripts that exert therapeutic activities. We have developed a specific ribozyme that can reprogram human telomerase reverse transcriptase (hTERT) RNA to induce transgene activity selectively in cancer cells that express the RNA. The ribozyme-mediated triggering of the transgene expression was accomplished via a high-fidelity trans-splicing reaction with the targeted residue in the hTERT expressing cells. The ribozyme also induced cytotoxic activity in various hTERT expressing cancer cells, hence selectively retarding the growth of those cells. Efficient and specific cell regression was also detected with ganciclovir (GCV) treatment only in hTERT-positive cancer cells, which were established to express stably the specific ribozyme that contains the herpes simplex virus thymidine kinase (HSV-tk) gene. Tissue-specific expression of the ribozyme could further augment the target specificity of the ribozyme. Importantly, we observed efficient regression of tumors with GCV treatment in mice that had been inoculated subcutaneously with hTERT-positive cancer cells that stably expressed the specific ribozyme that contains HSV-tk. These results suggest that the hTERT RNA-targeting trans-splicing ribozyme could be a powerful agent for tumor targeted specific gene therapy. PMID- 16040279 TI - Naming ability after tailored left temporal resection with extraoperative language mapping: increased risk of decline with later epilepsy onset age. AB - Standard temporal resection in the left hemisphere carries the risk of postoperative naming ability decline, especially with later epilepsy onset age/absence of hippocampal sclerosis. Language mapping has been performed routinely at some centers to minimize postoperative primary language impairment, but its effect on changes in naming performance has not been explored. This study examined naming outcome in 24 patients with nonlesional epilepsy who had left temporal resection after extraoperative language mapping. The mean decline in Boston Naming Test (BNT) score was 7.8, and 13 (54%) patients had a BNT decline greater than the Reliable Change Index. Simple correlations found significant relationships between BNT score decline and: later onset age, higher preoperative BNT score, and resection of isolated language sites. A multiple regression analysis showed that onset age was the best predictor of BNT decline. Although naming ability in patients with early onset age is stable with language mapping, there is still a risk of decline for those with later onset age. PMID- 16040280 TI - Respiratory responses of the air-breathing fish Hoplosternum littorale to hypoxia and hydrogen sulfide. AB - The present study analyzes the respiratory responses of the neotropical air breathing fish Hoplosternum littorale to graded hypoxia and increased sulfide concentrations. The oxygen uptake (VO2), critical O2 tension (PcO2), respiratory (fR) and air-breathing (fRA) frequencies in response to graded hypoxia were determined for fish acclimated to 28 degrees C. H. littorale was able to maintain a constant VO2 down to a PcO2 of 50 mm Hg, below which fish became dependent on the environmental O2 even with significant increases in fR. The fRA was kept constant around 1 breath h(-1) above 50 mm Hg and increased significantly below 40 mm Hg, reaching maximum values (about 4.5 breaths h(-1)) at 10 mm Hg. The lethality to sulfide concentrations under normoxic and hypoxic conditions were also determined along with the fRA. For the normoxic fish the sulfide lethal limit was about 70 microM, while in the hypoxic ones this limit increased to 87 muM. The high sulfide tolerance of H. littorale may be attributed to the air breathing capability, which is stimulated by this compound. PMID- 16040281 TI - Comparison of muscarinic receptor- and beta-adrenoceptor-mediated vasorelaxation between euthyroid and acute hyperthyroid rats. AB - Hyperthyroidism was induced by subcutaneous injections of L-thyroxine (T4) (0.5 mg/kg/day) for 3 days in order to investigate the effects of acute hyperthyroidism on the vasorelaxing responses to isoprenaline and acetylcholine in isolated rat aortae. In the aortae, there was no significant difference in isoprenaline-induced relaxation between hyperthyroid and control rats, however acetylcholine-induced relaxation was significantly greater in hyperthyroid rats than in control rats. N(G)-nitro-L-arginine (L-NOARG), an inhibitor of nitric oxide (NO) synthase, reduced isoprenaline- and acetylcholine-induced relaxations in both hyperthyroid and control rats and in the presence of L-NOARG no significant difference in the acetylcholine-induced relaxation was seen between the two groups of rats. Indomethacin, a cyclo-oxygenase inhibitor, had no significant influence on both isoprenaline- and acetylcholine-induced relaxations in both control and hyperthyroid rats. 17-Octadecynoic acid (17-ODYA), a cytochrome P-450 mono-oxygenase inhibitor, reduced the both isoprenaline- and acetylcholine-induced relaxation in both hyperthyroid and control rats, and acetylcholine-induced relaxation was still greater in hyperthyroid rats than in control rats. These results indicate that an acute hyperthyroidism significantly enhances muscarinic receptor- but not adrenoceptor-mediated relaxations of the aortae and L-NOARG abolished an enhancement by acute hyperthyroidism of muscarinic receptor-mediated relaxation, suggesting that the effects may be due to an alteration in muscarinic receptor-mediated NO systems of the aortae at early stage of hyperthyroidism. PMID- 16040282 TI - Medical bioremediation: prospects for the application of microbial catabolic diversity to aging and several major age-related diseases. AB - Several major diseases of old age, including atherosclerosis, macular degeneration and neurodegenerative diseases are associated with the intracellular accumulation of substances that impair cellular function and viability. Moreover, the accumulation of lipofuscin, a substance that may have similarly deleterious effects, is one of the most universal markers of aging in postmitotic cells. Reversing this accumulation may thus be valuable, but has proven challenging, doubtless because substances resistant to cellular catabolism are inherently hard to degrade. We suggest a radically new approach: augmenting humans' natural catabolic machinery with microbial enzymes. Many recalcitrant organic molecules are naturally degraded in the soil. Since the soil in certain environments - graveyards, for example - is enriched in human remains but does not accumulate these substances, it presumably harbours microbes that degrade them. The enzymes responsible could be identified and engineered to metabolise these substances in vivo. Here, we survey a range of such substances, their putative roles in age related diseases and the possible benefits of their removal. We discuss how microbes capable of degrading them can be isolated, characterised and their relevant enzymes engineered for this purpose and ways to avoid potential side effects. PMID- 16040283 TI - Activation of NMDA receptors prevents excessive metabolic decrease in hypoxic rat pups. AB - We tested the hypothesis that glutamate NMDA receptors may help maintain metabolic rate and body temperature during acute or chronic hypoxic exposure in newborn rats. We recorded ventilation, metabolism ((.)V(O(2)) -- ((.)V(CO(2)) and rectal temperature, under normoxia, acute hypoxia (30 min -- 12% O(2)), or following 10 days of chronic hypoxia, in 10 days old male and female rats, receiving saline i.p. injection or the NMDA receptor antagonist MK-801. Acute hypoxia decreased rectal temperature and metabolism, and increased ventilation, and (.)V(E)/((.)V(O(2) and (.)V(E)/((.)V(CO(2) to the same extent in males and females. MK-801 injection amplified the metabolic decrease under acute (in males and females) and chronic (in males) hypoxia, prevented the increase of minute ventilation, while (.)V(E)/((.)V(O(2) or (.)V(E)/((.)V(CO(2)remained constant. Hence, NMDA glutamate receptors help to maintain metabolic rate, minute ventilation and body temperature at a determined level in acute (males and females) and chronic hypoxia (males only). PMID- 16040284 TI - Levofloxacin- versus metronidazole-based rescue therapy for H. pylori infection in Japan. AB - BACKGROUND: The ideal second-line treatment regimens for Helicobacter pylori infection may differ between the areas, countries and races. AIM: The aim was to confirm which was the better regimen for second-line therapy after treatment failure with a standard triple therapy in Japan, a high dosage of levofloxacin- or metronidazole-based therapy. PATIENTS: Sixty outpatients with persistent H. pylori infection after a standard triple therapy were enrolled in this prospective, open-label and randomised trial. METHODS: The subjects were randomly administered levofloxacin (300 mg b.d.)- or metronidazole (500 mg b.d.)-based therapy with lansoprazole (30 mg b.d.) and amoxicillin (1000 mg b.d.) for 7 days, and the cure rates and side effects were analysed. Antimicrobial susceptibility was also examined before second-line therapy using the E-test. RESULTS: Good compliance was obtained without severe side effects in both the groups except for two patients. The cure rates, expressed as intention-to-treat and per-protocol analyses, respectively, were 70.0 and 72.4% in the levofloxacin group, and 96.7 and 100% in the metronidazole group. Each regimen often overcame even clarithromycin-resistant strains. CONCLUSION: Metronidazole-based triple therapy is recommended as second-line therapy in Japan, and levofloxacin-based therapy can be an alternative treatment option. PMID- 16040285 TI - The relative contributions of lean tissue mass and fat mass to bone density in young women. AB - Although obesity is associated with increased risk of many chronic diseases including cardiovascular disease, diabetes, hypertension, and cancer, there is little evidence to suggest that obesity increases risk of osteoporosis. In fact, both weight and body mass index (BMI) are positive predictors of bone mass in adults, suggesting that those who are overweight or obese may be at lower risk of osteoporosis. However, recent evidence suggests that in children and adolescents, obesity may be associated with lower rather than higher bone mass. To understand the relation of fat mass to bone mass, we examined data gathered from an ethnically diverse group of 921 young women, aged 20-25 years (317 African Americans, 154 Asians, 322 Caucasians, and 128 Latinas) to determine how fat mass (FM) as well as lean tissue mass (LTM) is associated with bone mass. Bone mass, FM, and LTM were measured using dual energy X-ray absorptiometry (GE Lunar Corp, Madison, WI). Bone mass was expressed as bone mineral density (BMD; g/cm2) and bone mineral apparent density (BMAD; g/cm3) for the spine and femoral neck, and as BMD and bone mineral content (BMC; g) for the whole body. Regression techniques were used to examine the following: (1) in separate equations, the associations of LTM and FM with each bone mass parameter; and (2) in the same equation, the independent contributions of LTM and FM to bone mass. LTM and FM were positively correlated with BMD at all skeletal sites. When the contributions of FM and LTM were examined simultaneously, both FM and LTM continued to be positively associated with bone mass parameters but the effect of FM was noted to be smaller than that of LTM. We conclude that in young women, LTM has a greater effect than fat mass on bone density per kg of tissue mass. PMID- 16040286 TI - Four-color flow cytometry shows strong concordance with bone marrow morphology and cytogenetics in the evaluation for myelodysplasia. AB - The ability of 4-color flow cytometry (FC) to help identify myelodysplastic syndromes (MDSs) was evaluated in 124 bone marrow aspirates from unselected patients with unexplained cytopenias and/or monocytosis. The morphologic features of bone marrow aspirate smears were correlated with FC and cytogenetic findings blindly, and patterns of antigen expression were compared with patterns seen in nonneoplastic and normal marrow specimens. Of 124 cases, 58 (46.7%) had definitive FC abnormalities ("flow-abnormal"), 19 cases (15.3%) had mild FC abnormalities of indeterminate significance, and 47 cases (37.9%) had essentially normal FC. Highly significant differences were identified between the flow abnormal group and other groups in mean myeloid blast percentages and numbers of abnormal antigens expressed, even when the analysis was limited to cases with fewer than 5% myeloid blasts. Strikingly, flow-abnormal cases constituted 50 (89%) of the 56 morphologically abnormal cases and 31 (94%) of the 33 cytogenetically abnormal cases, demonstrating the strong concordance of FC identified antigenic abnormalities with morphologic features and cytogenetics in the evaluation of patients with unexplained cytopenias. PMID- 16040287 TI - Follicular origin of a subset of CD5+ diffuse large B-cell lymphomas. AB - Most follicular lymphomas (FLs) have a phenotype consistent with the origin from CD5-, CD10+, bcl-6+ follicular center cells and can progress to diffuse large B cell lymphoma (DLBCL). CD5 is expressed in about 10% of DLBCLs, showing prognostic value, whereas expression is rare in FL. We present 6 cases with coexisting features of CD5+ FL and CD5+ DLBCL, supporting a follicular origin for some CD5+ DLBCLs. The follicular areas showed a meshwork of CD21+ follicular dendritic cells that were lacking in the DLBCL areas. All cases showed a clonal CD19+, CD20+, CD5+, and CD10+ population in both follicular and diffuse areas. Molecularly, 4 of 6 cases demonstrated immunoglobulin heavy chain rearrangements and 1 case, a bcl-2/immunoglobulin heavy chain gene rearrangement. Somatic hypermutations were high in all 4 cases, in keeping with their germinal center origin. Four of five patients died of disease within 42 months, consistent with the proposed prognostic value of CD5 expression in DLBCL. Our data describe an aggressive variant of CD5+ FL suggesting the follicular origin of some CD5+ DLBCLs. PMID- 16040288 TI - Low blast count myeloid disorders with Auer rods: a clinicopathologic analysis of 9 cases. AB - Auer rods are a hallmark of acute myeloid leukemia but occasionally are seen in myelodysplastic syndromes (MDSs) or chronic myelomonocytic leukemia, rarely in cases with fewer than 5% blasts. The significance of this finding is unclear. We report 9 cases of this unusual phenomenon. All patients had cytopenias, isolated to a single lineage in 4. Circulating blasts were present in 8 cases (rare to 2.5%). Bone marrow blasts ranged from 0.4 to 4.9%; 1% to 32% of blasts contained Auer rods. There were variable degrees of dysplasia; 1 case closely mimicked refractory anemia with ringed sideroblasts. Cytogenetic studies in 8 cases showed clonal changes in 4. In 5 patients, acute myelogenous leukemia (AML) developed 6, 6, 5, 13, and 24 months after diagnosis; the patients subsequently died. Three patients died at 1, 1, and 8 months without progression to AML, and only 1 was alive at 10 months. MDSs with fewer than 5% blasts and Auer rods seem to be a heterogeneous group, but rapid progression to death or AML in most cases suggests that Auer rods signify an aggressive biology in MDSs with a low blast count. PMID- 16040289 TI - Flow cytometric immunophenotyping of adult T-cell leukemia/lymphoma using CD3 gating. AB - Adult T-cell leukemia/lymphoma (ATLL) is a lymphoproliferative neoplasm of helper T lymphocytes caused by human T-cell leukemia virus type-1 (HTLV-1). The disease was first described in Kyushu, in southwestern Japan, and most frequently occurs in endemic areas, such as Japan, the Caribbean basin, West Africa, Brazil, and northern Iran. ATLL is essentially a disease of adults, characterized clinically by generalized lymphadenopathy, hepatosplenomegaly, skin lesions, and hypercalcemia. The prognosis of most patients is quite poor, with a median survival time of only 13 months, even if multiagent combination chemotherapy is given. In the present study, flow cytometric immunophenotyping with CD3 gating was performed on 30 samples from 26 patients who had been given a diagnosis of ATLL. The records of these patients also were reviewed retrospectively. In 14 of the 30 samples, an abnormal CD3(low) T-cell population was distinguishable from the normal T-cell populations by flow cytometric analysis. Herein we report a novel strategy for flow cytometric immunophenotyping of ATLL facilitated by CD3(low) gating. PMID- 16040290 TI - Acetic acid-zinc-formalin: a safe alternative to B-5 fixative. AB - B5 fixation achieves superior morphologic detail. However, environmental concerns have led to labor-intensive and costly requirements for disposal of mercury containing fixatives. We performed a blinded prospective study to find a safe, mercury-free alternative to B5. Morphology was evaluated with 6 fixatives, including B5, in a blinded fashion. Acetic acid-zinc-formalin (AZF) was selected for further evaluation of immunohistochemistry, in situ hybridization, and molecular analysis. AZF fixation resulted in overall staining and morphologic detail comparable to B5 and achieved equivalent or superior antigen preservation for immunohistochemical studies. Strong signal intensity was achieved with in situ hybridization, and DNA amplification could be successfully performed. AZF allows greater flexibility in fixation times, decreases decalcification time, and eliminates labor-intensive steps required for B5 processing. PMID- 16040291 TI - Serum free light chain (FLC) measurement can aid capillary zone electrophoresis in detecting subtle FLC-producing M proteins. AB - We hypothesized that using a free light chain (FLC) assay as an adjunct to capillary zone electrophoresis (CZE) could improve detection of lymphoplasmacytic processes. We prospectively studied 1,003 consecutive serum samples submitted for routine protein electrophoresis and/or immunofixation electrophoresis by CZE and FLC. Samples from patients previously characterized as having M proteins were excluded. Protein electrophoresis was read by a pathologist unaware of the FLC results. Sixteen cases revealed an abnormal free kappa/lambda ratio in which CZE did not demonstrate an M protein. Nine cases of B-lymphocyte or plasma cell proliferative processes were detected by an abnormal free kappa/lambda ratio in which CZE did not demonstrate an M protein. Cases with low free kappa/lambda ratios included 1 chronic lymphocytic leukemia (CLL), 1 IgM lambda with aplastic anemia, and 1 lambda light chain myeloma. Cases with high free kappa/lambda ratios included 2 CLL, 1 lymphocytosis (possibly early CLL), 1 kappa light chain myeloma, 1 atypical lymphoma with neuropathy, and 1 nonsecretory myeloma. Addition of the free kappa/lambda ratio to CZE increases the yield of lymphocyte and plasma cell proliferative processes detected by 56%. PMID- 16040292 TI - Performance characteristics of an HPLC assay for urinary albumin. AB - Microalbuminuria is a marker of diabetic nephropathy and cardiovascular risk. Immunoassays underestimate the amount of intact albumin present in urine. The purpose of this study was to evaluate a new urinary albumin assay that uses size exclusion high-performance liquid chromatography (HPLC). We determined the limit of detection, linearity, imprecision, a comparison with an immunoturbidimetric assay, and pediatric and adult reference intervals. The limit of detection was 3.4 mg/L. The assay was linear from 4 to 240 mg/L. Total imprecision was less than 10% from 16 to 206 mg/L. Comparison of the albumin/creatinine ratio by HPLC with an immunoturbidimetric method showed positive proportional bias, which decreased with increasing concentrations of albumin. Nonparametric reference intervals were 22 to 250 mg/g for girls, 20 to 130 mg/g for boys, 14 to 62 mg/g for women, and 10 to 37 mg/g for men. This HPLC assay for urinary albumin shows acceptable performance and quantifies albumin species that are not detected by immunoassay. Separate reference intervals for children and adults seem necessary. PMID- 16040293 TI - Studies of the inhibition of serum pseudocholinesterase activity in vitro by commonly used drugs. AB - We studied the effect of 17 commonly used drugs, including prescription and over the-counter medications, on the activity of serum pseudocholinesterase (PCE) in vitro. Normal pooled human serum was incubated for 120 minutes at 37 degrees C with therapeutic serum concentrations of prescription and over-the-counter drugs, and the postincubation PCE activity was measured. Morphine, quinidine, and thioridazine depressed PCE activity by more than 5% while no or negligible effect was noted following incubation with acetaminophen, chlordiazepoxide, chlorpromazine, desipramine, doxepin, imipramine, methamphetamine, nortriptyline, phenobarbital, phenytoin, procainamide, salicylic acid, theophylline, and valproic acid. Depression of PCE activity can prolong the half-life of coadministered agents with metabolism mediated by PCE. PMID- 16040294 TI - Effect of Brazilian, Indian, Siberian, Asian, and North American ginseng on serum digoxin measurement by immunoassays and binding of digoxin-like immunoreactive components of ginseng with Fab fragment of antidigoxin antibody (Digibind). AB - We compared Brazilian, Indian, Siberian, Asian, and North American ginseng for potential interference with 3 digoxin immunoassays: fluorescence polarization (FPIA), microparticle enzyme (MEIA), and Tina-quant (Roche Diagnostics, Indianapolis, IN). We supplemented aliquots of a drug-free serum pool with ginseng extracts representing expected in vivo concentrations and overdose. We observed apparent digoxin-like immunoreactivity with FPIA, modest immunoreactivity with MEIA, and no apparent digoxin immunoreactivity with the Tina-quant with all ginsengs except Brazilian, which showed no immunoreactivity with any assay. When aliquots of serum pools prepared from patients receiving digoxin were supplemented with ginsengs, we observed falsely elevated digoxin values with FPIA, falsely lower digoxin values (negative interference) with MEIA, and no interference with the Tina-quant. Digoxin-like immunoreactive components of various ginsengs have moderate protein binding; monitoring free digoxin concentrations does not eliminate such interference. We also observed that Digibind (Burroughs Wellcome, Research Triangle Park, NC) can bind free digoxin like immunoreactive components of ginsengs; such effects can be monitored by measuring apparent free digoxin concentrations. Indian, Asian, and North American ginsengs interfere with serum digoxin measurement by FPIA and MEIA; the Tina quant is free of such interference. Digibind can bind free digoxin-like immunoreactive components of ginseng. PMID- 16040295 TI - Interobserver variability in thyroid fine-needle aspiration interpretation of lesions showing predominantly colloid and follicular groups. AB - We studied interobserver variability (IV) in the assessment of thyroid fine needle aspiration (FNA). We limited our cases to those showing predominantly colloid and follicular cell groups. Twenty cases of thyroid FNA diagnosed by 1 experienced cytopathologist were reviewed by 4 other cytopathologists who made their own diagnoses while unaware of the original diagnoses. Two cytopathologists then assessed the cytologic features of the 20 cases. IV was calculated for noncollapsed and collapsed diagnoses. Diagnoses and observer agreement were compared with cytologic features. There was little correlation among observers regarding the diagnosis of follicular "lesion" vs "neoplasm." IV was somewhat poor before data were collapsed to treatment recommendations (kappa = 0.35) but was relatively good when data were collapsed (kappa = 0.65). Cellularity, cyst change, and amount of colloid correlated with treatment recommendations; no specific features correlated with poor performance. Thyroid FNA shows good interobserver agreement in the diagnoses of lesions showing predominantly colloid or follicular cells (when collapsed). We speculate that IV is poor in some cases owing to difficulty assessing thin colloid, some lack of agreement regarding criteria for adequacy, and a possible "gray zone" that might exist with lesions showing colloid and abundant follicular cells. PMID- 16040296 TI - Paratesticular cysts with benign epithelial proliferations of wolffian origin. AB - Paratesticular cysts with benign epithelial proliferations (BEPs) are rare. Only 10 cases were found in a series of 431 paratesticular cysts and were classified as follows: cystadenoma, 5; papilloma, 2; and hamartoma, 3. Four cystadenomas showed multiple papillae lined by CD10+ epithelial cells with hyperchromatic nuclei. The remaining lesion showed areas with a microcystic, glandular, cribriform pattern, with small, benign glands without atypia. Urothelial papilloma presented BEPs with cytokeratin (CK) 7+ and CD10+ and CK20- umbrella like cells. The mural papilloma was lined by proliferative cylindrical cells exhibiting strong CK7 and CD10 expression. The 3 Wolffian hamartomas were characterized by strongly CD10+ epithelium surrounded by smooth muscle cells. The consistent CD10 expression in BEPs of paratesticular cysts suggests a Wolffian origin. The differential diagnosis of paratesticular cysts with BEP vs metastatic prostatic and primary borderline or malignant tumors is discussed. PMID- 16040297 TI - Lack of alkaline phosphatase activity predicts meningioma recurrence. AB - Meningiomas usually are benign intracranial tumors. However, some recur despite gross total resection, invade surrounding structures, or, rarely, metastasize. Reduced expression of the nonspecific tissue-type alkaline phosphatase (Pal) has been reported in high-grade meningiomas. To search for a predictor for recurrence, we studied Pal expression by histoenzymology in a series of 54 meningiomas with gross total removal. Pal expression was mostly altered in grades II and III meningiomas (P = .000014) and meningiomas with high MIB-1 values (P = .001). Pal expression correlated with cytogenetic data (P = .000033) and with recurrence (P = .0064); all tumors that recurred had abnormal Pal expression (13/13). In univariate analysis, Pal expression was the only variable correlated with recurrence-free survival (P = .035). Histochemical detection of Pal is a useful, easy, and low-cost technique to predict recurrence in meningiomas. PMID- 16040298 TI - Techniques and criteria in pathologic and forensic-medical diagnostics in sudden unexpected infant and perinatal death. AB - For each case of sudden infant and perinatal death, a full review of clinical and epidemiologic data and a complete necropsy study were performed according to the necropsy protocol devised by the Institute of Pathology, University of Milan, Milan, Italy (available at: http://users.unimi.it/~pathol/sids_e.html). Histopathologic examination of unexpected late fetal and neonatal death and SIDS cases allowed us to identify frequent alterations, mainly congenital, of the autonomic nervous system, modulating respiratory, cardiovascular, arousal, and upper digestive activities. The data and arguments presented herein provide a brief survey tending to open, rather than conclude, a far-reaching subject and to motivate medicolegal specialists and pathologists to perform more in-depth study. PMID- 16040299 TI - Colorectal intramucosal perikarya of ganglion cells. AB - It generally is believed that perikarya of ganglion cells in the human colorectum are confined to plexuses that lie deep to the mucosa, and that intramucosal perikarya are rare. We retrospectively reviewed 100 specimens from biopsies of normal and abnormal mucosa to further characterize intramucosal perikarya. The presence of intramucosal perikarya, their number, location, and grouping were recorded. Twenty-one specimens (21.0%) contained intramucosal perikarya. Intramucosal perikarya occurred throughout the colorectum in the muscularis mucosae or lamina propria and in normal mucosa, acute self-limited colitis, inflammatory bowel disease, cytomegalovirus-associated colitis, hyperplastic polyps, tubular adenomas, and high-grade intraepithelial neoplasia. In some specimens, intramucosal perikarya morphologically resembled microgranulomas or cytomegalovirus-infected cells. We demonstrated that intramucosal perikarya of ganglion cells are surprisingly common in normal and abnormal mucosa. Awareness of intramucosal perikarya is necessary to avoid confusion with microgranulomas or cytomegalovirus-infected cells. PMID- 16040300 TI - Analysis of intratumoral heterogeneity and amplification status in breast carcinomas with equivocal (2+) HER-2 immunostaining. AB - Fluorescence in situ hybridization (FISH) and immunohistochemical analysis for assessment of HER-2 status in breast carcinomas are discordant in a significant proportion of cases with equivocal (2+) immunostaining. To evaluate the role of intratumoral heterogeneity and degree of amplification, we performed additional HER-2 immunostains and FISH on tumor-bearing blocks from 20 invasive breast carcinomas with immunohistochemical scores of 2+ with gene amplification and in 18 cases without amplification. Of the amplified cases, 11 (55%) had a 3+ immunohistochemical score on at least 1 additional slide, 8 (40%) remained 2+, and 1 (5%) had a slide scored 1+. All cases rescored 3+ showed high-level amplification in original and repeated FISH; cases remaining 2+ had a heterogeneous FISH profile (low-level amplification or a mosaic mixture of high level amplified and nonamplified cells) in original and repeated FISH. Of nonamplified cases, 13 (72%) had a 1+ score on at least 1 additional slide, 4 (22%) remained 2+, and 1 (6%) had 1 slide scored 3+. In the nonamplified cases, 17 (94%) showed no amplification in repeated FISH. Significant intratumoral heterogeneity and minimal (low-level) HER-2 amplification account for many breast cancers with 2+ HER-2 protein expression. PMID- 16040301 TI - Overexpression of decoy receptor 3 in precancerous lesions and adenocarcinoma of the esophagus. AB - Overexpression of decoy receptor (DcR) 3 protein, a recently discovered member of the tumor necrosis factor receptor superfamily, was examined in 40 esophagogastrectomy specimens containing areas of Barrett esophagus (n = 27), low grade dysplasia (n = 27), high-grade dysplasia or carcinoma in situ (n = 22), and esophageal adenocarcinoma (EAC; n = 28) with immunohistochemical analysis. The results revealed significantly more overexpression of DcR3 in high-grade dysplasia or carcinoma in situ and EAC than in benign esophageal mucosa (both P < .0001), Barrett esophagus (both P < .001), and low-grade dysplasia (P < .01 and P = .033, respectively). Low-grade dysplasia also showed significant overexpression of DcR3 compared with benign esophagus (P < .05) but not with Barrett esophagus (P > .05). DcR3 overexpression seems to negatively correlate with the grade of EAC. Our results suggest that overexpression of DcR3 protein might aid in the diagnosis of high-grade dysplasia or carcinoma in situ and EAC and also might serve as a potential therapeutic target. PMID- 16040302 TI - Intralesional fibrous septum in chordoma: a clinicopathologic and immunohistochemical study of 122 lesions. AB - Intralesional fibrous septum (IFS) generally is considered a reactive tissue in chordoma; however, little is known about its significance. We studied 122 chordomas for IFS using immunohistochemical techniques and compared IFS and lobular growth patterns (LGPs) formed by IFS with clinicopathologic parameters. Seventy-nine tumors (64.8%) revealed IFS. However, IFS frequently was infiltrated and interrupted by tumor cells with increased expression of proteases; only 33 (42%) of 79 tumors had LGP. In non-skull base chordomas, IFS and LGP were associated with nuclear pleomorphism, a previously described prognostic indicator, mitosis, and the MIB-1 labeling index, indicating a role of IFS and LGP in tumor growth or progression. Paradoxically, patients without LGP tended to have a worse prognosis than those with LGP. We believe that IFS exerts diverse influences on chordoma; however, invasion of IFS leading to loss of the LGP indicates advanced stages of tumor development, possibly predicting an unfavorable prognosis in chordoma. PMID- 16040303 TI - Rabbit monoclonal antibodies: a comparative study between a novel category of immunoreagents and the corresponding mouse monoclonal antibodies. AB - Rabbit monoclonal antibodies (RabMAbs) represent a novel category of immunoreagents that may combine the best properties of both mouse monoclonal antibodies (MMAs) and of rabbit antisera. In the attempt to verify the performance of this new class of antibodies on paraffin-embedded tissue, RabMAbs against estrogen receptor, progesterone receptor, Ki-67, cyclin D1, CD3, CD5, CD23, and synaptophysin were tested on several tumor types as well as normal tissues. The results were compared with those obtained with classic MMAs against the same antigens. RabMAbs appear to offer increased sensitivity with no apparent loss of specificity. On routine use they permit higher working dilutions (5 to 10 times on average), allowing significant improvement in terms of laboratory efficiency. The robustness of RabMAbs is further proved by the fact that in some instances optimal staining can be obtained even without antigen retrieval. In consideration of the high performance observed, routine use of RabMAbs may contribute significantly to standardize diagnostic immunohistochemical procedures. PMID- 16040304 TI - Are sequence variations in the BK virus control region essential for the development of polyomavirus nephropathy? AB - BK virus replication is regulated by the noncoding control region (NCCR); major NCCR rearrangements could modify the strength of viral replication, having a role in the development of polyomavirus-associated nephropathy (PAN). Urine (n = 34), blood (n = 32), and renal biopsy samples (n = 13) from 5 transplant recipients with PAN underwent nested polymerase chain reaction to search for the NCCR region. Sequence analysis was performed on all NCCR fragments obtained. Decoy cells were evaluated semiquantitatively in urine and PAN staged in renal biopsy specimens; the results were related to the presence and type of NCCR sequence variations. Major NCCR rearrangements were found in urine (9/75 [12%]), blood (7/30 [23%]), and renal biopsy (4/15 [27%]) samples in 3 cases; 2 cases had only unrearranged strains. Neither the detection and number of decoy cells nor the PAN stage were related to the specific type of NCCR sequence rearrangements. NCCR rearrangements do not seem essential for the development of PAN. PMID- 16040305 TI - The relevance of psychoanalysis to an understanding of terrorism. PMID- 16040306 TI - 20 Years is not long enough. AB - The author presents some Latin American sociopolitical vicissitudes exemplified by Argentina, where she lives and where she trained and practices as a psychoanalyst. The exposition is based on the impact that her experience with two patients, Ana and Juana, had on her, and is presented in the form of clinical vignettes. The author reflects clinically and technically on the transference and countertransference and on the ways in which self-analysis enabled her to distinguish between the countertransference related to the patient and that related to the psychoanalyst. Finally, the author discusses the traumatic effects of 'the human condition', 'social violence' and 'Evil', referring specifically to the 'repetitive trauma' individuals experience under the globalization of terror and to the use of mechanisms of disavowal that result in serious splitting. The author confronts the reader with totalitarian terror as something that attacks and destroys the main constitutive characteristic of human beings, namely, their ability to think, remarking that H. Arendt is the one who speaks about 'radical evil' as 'the banality of Evil'. The author addresses the question of whether by tempering aggression and organizing levels of symbolization 'words' might prevent the emergence of 'pure jouissance' and be more powerful and significant than violence, overriding it. PMID- 16040307 TI - 'Yes, we have bananas!'. AB - This paper takes as its starting point 'identifying markers of Latin American psychoanalysis'. The authors see this theme as a compromise between two diverging approaches. On one hand, the conference at which the content was presented provided the opportunity for Latin American psychoanalytical thinking to be discussed and, as a reward to the best papers, to be published. On the other, both the conference and the reward are more indicative of the observer's biases than of the object's peculiarities. The authors consider that the conference mistakenly focused on the search for minor differences (secondary identifications) instead of looking for invariances (identities). Considering that uncertainty, pluralism and complexity are issues relevant to the whole of psychoanalysis in its current stage of development as a scientific discipline, the authors think that treating these elements as 'identifying markers of Latin American psychoanalysis' is evidence of prejudice brought about by a coloniser colonised relationship. To develop their argument, the authors discuss a paper on editorial criteria by Tuckett as a clinical case, and use an episode of Brazilian history as further illustration. They address issues such as conformity to cultural patterns; the search for certainties and proofs; the domination of some groups over others; and editorial power-more specifically, its influence on the acceptance or rejection of scientific ideas. These issues have distracted attention away from the fundamentals of psychoanalysis by introducing other, extraneous aims. Publication plays a key role in feeding a possibly vicious circle wherein only a small proportion of scientific contributions manages to reach a wide audience--exactly those contributions that conform to established patterns. The act of publication turns that portion into official knowledge, while unpublished ideas become increasingly excluded. PMID- 16040308 TI - New interpretative styles: progress or contamination?: psychoanalysis and phenomenological psychopathology. AB - Psychoanalysis has started to recoup, often quite implicitly, a more phenomenological stance, ever since psychoanalysts have started working with borderline and psychotic patients. As many of these patients have commonly been through traumatic experiences, psychoanalysts have been using an approach that questions the role of traditional psychoanalytical interpretation and pays more attention to the patient's inner conscious experiences; this approach is characteristic of a specific form of contemporary psychiatry: phenomenological psychopathology, founded by Karl Jaspers in 1913 and developed into a form of psychotherapy by Ludwig Binswanger, with his Daseinsanalyse. If what we could call a phenomenological 'temptation' has been spreading over psychoanalysis, so too has a psychoanalytical 'temptation' always been present in phenomenological psychopathology. In fact, even though this branch of psychiatry has led us towards a deeper understanding of the characteristics of psychotic being-in-the world, its therapeutic applications have never been adequately formalised, much less have they evolved into a specific technique or a structured psychotherapeutic approach. Likewise, phenomenological psychotherapy has always held an anaclitic attitude towards psychoanalysis, accepting its procedures but refusing its theoretical basis because it is too close to that of the objectifying natural sciences. Psychoanalytic 'temptation' and phenomenological 'temptation' can thus be considered as two sides of the same coin and outline a trend in psychoanalytic and phenomenological literature which points out the fundamental role of the patient's inner conscious experiences in the treatment of borderline and psychotic patients. PMID- 16040309 TI - Trauma, narcissism and the two attractors in trauma. AB - In this paper, the author sets out to distinguish anew between two concepts that have become sorely entangled--'trauma' and 'narcissism'. Defining 'narcissism' in terms of an interaction between the selfobject and the self that maintains a protective shield, and 'trauma' as attacks on this protective shield, perpetrated by bad objects, he introduces two attractors present in trauma--'the hole attractor' and the structure enveloping it, 'the narcissistic envelope'. The hole attractor pulls the trauma patient, like a 'black hole', into a realm of emotional void, of hole object transference, devoid of memories and where often in an analyst's countertransference there are no reverberations of the trauma patient's experience. In the narcissistic envelope, on the other hand, motion, the life and death drive and fragments of memory do survive. Based on the author's own clinical experience with Holocaust survivors, and on secondary sources, the paper concludes with some clinical implications that take the two attractors into account. PMID- 16040310 TI - 'Projective transidentification': an extension of the concept of projective identification. AB - Questions about the concept of projective identification still persist. The author presents the following hypotheses: Klein's traditional view and Bion's extension and revision of it can be thought of as occupying a continuum in reverse. He postulates that Bion's concept of communicative intersubjective projective identification (which the author renames 'projective transidentification') is primary and inclusive of Klein's earlier unconscious, omnipotent, intrapsychic mode but includes Bion's 'realistic' communicative mode as well. The author hypothesizes, consequently, that intersubjective projective identification constitutes both the operation of an unconscious phantasy of omnipotent intrapsychic projective identification solely within the internal world of the projecting subject--in addition to two other processes: conscious and/or preconscious modes of sensorimotor induction, which would include signaling and/or evocation or prompting gestures or techniques (mental, physical, verbal, posturing or priming) on the part of the projecting subject; followed by spontaneous empathic simulation in the receptive object of the subject's experience in which the receptive object is already inherently 'hard-wired' to be empathic with the prompting subject. PMID- 16040311 TI - Pentheus rather than Oedipus: on perversion, survival and analytic 'presencing'. AB - Following an introductory review of the main developments in the psychoanalytic thinking on perversion, the author focuses on her own understanding of perversion and its treatment, based on the psychoanalytic treatment of patients with severe sexual perversions. This paper uses the term 'autotomy' (borrowed from the field of biology) to describe perversion formation as an 'autotomous' defence solution involving massive dissociative splitting in the service of psychic survival within a violent, traumatic early childhood situation; thus, a compulsively enacted 'desire for ritualised trauma' ensues. The specific nature of the perverse scenario embodies the specific experiential core quality of the traumatic situation. It is an actual repetition in the present of the imprint of a past destructive experience which is pre-arranged and stage-managed; it thus encounters haunting scenes of dread or psychic annihilation while, at the same time, controlling, sanitising and disavowing them. Hence, the world of severe perversion is no longer oedipal, but rather the world of Pentheus, Euripides's most tragic hero--a world dominated by a mixture of a mother's madness, devourment, destruction and rituals of desire. According to this view, the (difficult) psychoanalytic treatment of perversion focuses on patient-analyst interconnectedness--brought about by the analyst's 'givenness to being present' or 'presencing'--at a deep, primary level of contact and impact (the emphasis being on the ontological dimension of experience). This evolving therapeutic entity creates and actualises a new, alternative experiential-emotional reality within the pervert's alienated world, eventually generating a change in the perverse essence. The author illustrate this approach with three clinical vignettes. PMID- 16040312 TI - Approaching countertransference in psychoanalytical supervision: a qualitative investigation. AB - A recent examination of the literature concerning countertransference and its developments reveals its clinical usefulness in different psychoanalytical cultures. Nevertheless, a shortage of publications is apparent with respect to its approach in supervision. The authors' aim in this study was to examine the concepts of transference and countertransference and how countertransference is approached in supervision, in the training of candidates, at an institute of psychoanalysis belonging to a society affiliated to the International Psychoanalytical Association (IPA). Qualitative research was carried out, interviewing supervisors and supervisees. Through analysis of the material acquired, the authors classified the data into initial, intermediate and final categories. The principal findings were subdivided into three categories: the concepts of transference and countertransference, psychoanalytical listening and the complementarity of the phenomena, and the approach to countertransference. The concepts of transference and countertransference predominantly used by those interviewed are based on the totalistic outlook. Countertransference in supervision has been approached in a more direct and objective way when compared with the previous period, although great care is taken to delimit the boundaries between supervision and personal analysis. The main aim of supervision is to broaden comprehension and to deepen the interpretations directed towards the patient. These findings suggest that the evolution of the concept of countertransference in different psychoanalytical cultures and developments in the analytical field are contributing to this change. PMID- 16040313 TI - No reconciliation, but self-searching in the sense of rapprochement: Hillel Klein's Holocaust research in Germany 40 years after. AB - The paper is written from a personal perspective and in conjunction with the author's participation in editing Hillel Klein's book Uberleben und Versuche der Wiederbelebung-Psychoanalytische Studien mit Uberlebenden der Shoah und mit ihren Familien in Israel und in der Diaspora [Survival and trials of revival- Psychodynamic studies of Holocaust survivors and their families in Israel and the diaspora] which appeared posthumously in 2003. The manuscript was originally written in English during the first half of the 1980s. It consists of a revised version of his contributions to psychoanalytic Holocaust research, as well as further observations and a number of autobiographical features. It suggests that we see the symptom which relates to the traumatic past less as the sign of a pathology, and instead, through the linking of drives and objects, more as a sign of hope. The hope is that revival begins in recovering the world that was lost in the Holocaust, the world from before, which, as torn as it was, he still referred to as the 'intact world'. He regarded rapprochement as one of the main therapeutic tools. Rapprochement does not build up absolutes, but leaves from the quantitative point of view room for encounter. PMID- 16040314 TI - The Emperor's clothes: some serious problems in psychoanalytic training. AB - The author discusses some of the key problems in psychoanalytic training, in particular those problems that stem from the power differential between training analysts and students in training. One effect of this differential can be that some students feel a pressure to comply with their teachers and supervisors, even their training analyst, in ways that can be seriously detrimental to their development. Further, when something goes wrong in a student's training, how is this to be viewed by those in charge of the training? Also, how are complaints dealt with? Is sufficient weight given to external reality? Too often training analysts, and training committees, get into pathologising a student in a process that should be recognised as 'wild analysis in committee', rather than considering more carefully the external realities that may be affecting a student's progress in the training. This 'analysis' in committee should never be allowed. There is an urgency for immediate changes to be made in psychoanalytic training so that the problems discussed, with more care being taken, should be prevented from happening. Too often, however, an institutional resistance to change dominates discussions in committee, and in society meetings, with the result that little or no change takes place even after years of debate. PMID- 16040315 TI - Missing what was true: problems of seeing and knowing in Henry James's The wings of the dove. AB - The author examines a central theme in this late novel by Henry James in relation to current psychoanalytic ideas that link the Oedipus complex with the child's developing perception of reality (both psychic and external), specifically through the experience of seeing and being seen. Britton visualises the oedipal triangle as a psychic structure through which the child may achieve recognition not only of its parents' sexual relationship, from which it is excluded, but also of itself being observed by one parent while the child is with the other. Thus, it both observes and is observed. The differing perspectives achieved--of subjectivity and objectivity--promote the perception of objective reality, as the world of relationships grows and becomes more complex. James captures with great subtlety and penetration the experience of three characters living out a symbolic oedipal relationship in which the truth is evaded or perverted. A young couple in love exploit the situation of a dying heiress whose vulnerability is intensified by her reluctance to acknowledge the truth about their relationship. At the same time, she shrinks from the gaze of others and consigns herself to isolation and ultimate despair. The author presents three significant scenes in which seeing and being seen are central to the development. In each, the dying woman is forced to face, if momentarily, her exclusion from the sexual relationship. Increasingly this connects with her approaching death--but also with the anguished recognition that the couple have cruelly befriended her only to betray her. It is suggested that James's late style and novelistic technique require the reader to tolerate confusion and uncertainty. As the perspective shifts from one protagonist to another, we ourselves are in danger of 'missing what is true' in this characteristic Jamesian scenario, where relationships are gradually perverted by manipulation, evasion and lies. In psychoanalytic theory, this would represent a failure to work through the oedipal situation, where the struggle of the child to face reality is met by a parental relationship that is too weak or too perverse to contain the pain and conflict. PMID- 16040316 TI - Psychoanalysis and psychotherapy: a long and troubled relationship. AB - The extended, inconclusive debate over distinguishing psychoanalysis and psychotherapy has been muddied by two underlying issues. The scientific identity that psychoanalysis claimed, leading it to affirm that it was more than a treatment modality, was in conflict with the actual therapeutic mission it assumed. Psychoanalysis was further hampered in those discussion by its internal conflicts over doctrinal purity; deviations for psychotherapeutic ends were vulnerable to the charge of dissidence. More recently, the debate has been clouded by the fact that newer candidates, by and large, can anticipate careers primarily as psychotherapists, driving a wedge between generations within institutes. The course of the debate and the problems encountered in it are affected by the formal relations between the psychoanalytic establishment and the health-care industry, including government agencies. In the long run, it appears to make little difference whether psychoanalysis is officially recognized as a mental-health treatment, as in Germany, or attempts to maintain its independence, as in the UK. Finally, as the debate appears to be winding down, the fate of dynamic psychotherapy is also in the balance. If in the past psychoanalysis seemed at risk of losing its specific identity, today dynamic psychotherapy is in danger as well. PMID- 16040317 TI - On: editors' note for the Analyst at work feature. PMID- 16040320 TI - The influence of vaginally applied imiquimod on the course of Chlamydia trachomatis serovar D infection in a murine model. PMID- 16040321 TI - Opportunities to reduce overuse of antibiotics for perinatal group B streptococcal disease prevention and management of preterm premature rupture of membranes. AB - OBJECTIVE: To identify opportunities to reduce overuse of antibiotics for prevention of perinatal group B streptococcal (GBS) disease and management of preterm premature rupture of membranes (pPROM). METHODS: An anonymous written questionnaire was sent to each of 1031 Fellows of the American College of Obstetricians and Gynecologists, and the responses were subjected to statistical analysis. RESULTS: Among those of the 404 respondents who saw obstetric patients in 2001, most (84%) screened for GBS colonization, and 22% of these prescribed prenatal antibiotics to try to eradicate GBS colonization. Of the 382 respondents (95%) who prescribed antibiotics for pPROM, 36% continued antibiotics for more than 7 days despite negative results from GBS cultures collected before initiation of treatment. Having more years of clinical experience (adjusted odds ratio (OR) 3.0, 95% confidence interval (CI) 1.5 to 6.2), working in a non academic setting (adjusted OR 2.7, 95% CI 1.0 to 6.9), and prescribing antibiotics prenatally for GBS colonization (adjusted OR 2.0, 95% CI 1.1 to 3.4) were associated with prescribing prolonged antibiotics for pPROM. CONCLUSION: Prenatal antibiotic treatment for GBS colonization and prolonged antibiotic treatment for pPROM contribute to overuse of antibiotics in obstetrics. PMID- 16040322 TI - Listeriosis prevention knowledge among pregnant women in the USA. AB - BACKGROUND: Listeriosis is a food-borne disease often associated with ready-to eat foods. It usually causes mild febrile gastrointestinal illness in immunocompetent persons. In pregnant women, it may cause more severe infection and often crosses the placenta to infect the fetus, resulting in miscarriage, fetal death or neonatal morbidity. Simple precautions during pregnancy can prevent listeriosis. However, many women are unaware of these precautions and listeriosis education is often omitted from prenatal care. METHODS: Volunteer pregnant women were recruited to complete a questionnaire to assess their knowledge of listeriosis and its prevention, in two separate studies. One study was a national survey of 403 women from throughout the USA, and the other survey was limited to 286 Minnesota residents. RESULTS: In the multi-state survey, 74 of 403 respondents (18%) had some knowledge of listeriosis, compared with 43 of 286 (15%) respondents to the Minnesota survey. The majority of respondents reported hearing about listeriosis from a medical professional. In the multi-state survey, 33% of respondents knew listeriosis could be prevented by not eating delicatessen meats, compared with 17% in the Minnesota survey (p=0.01). Similarly, 31% of respondents to the multi-state survey compared with 19% of Minnesota survey respondents knew listeriosis could be prevented by avoiding unpasteurized dairy products (p=0.05). As for preventive behaviors, 18% of US and 23% of Minnesota respondents reported avoiding delicatessen meats and ready-to-eat foods during pregnancy, whereas 86% and 88%, respectively, avoided unpasteurized dairy products. CONCLUSIONS: Most pregnant women have limited knowledge of listeriosis prevention. Even though most respondents avoided eating unpasteurized dairy products, they were unaware of the risk associated with ready-to-eat foods. Improved education of pregnant women regarding the risk and sources of listeriosis in pregnancy is needed. PMID- 16040323 TI - Knowledge, attitudes, and reported practices among obstetrician-gynecologists in the USA regarding antibiotic prescribing for upper respiratory tract infections. AB - BACKGROUND: Knowledge, attitudes, and practices regarding antibiotic prescribing for upper respiratory tract infections (URIs) have not been well described among obstetrician-gynecologists (OB/GYNs). This information is useful for determining whether an OB/GYN-specific program promoting appropriate antibiotic use would significantly contribute to the efforts to decrease inappropriate antibiotic use among primary care providers. METHODS: An anonymous questionnaire asking about the treatment of URIs was sent to 1031 obstetrician-gynecologists. RESULTS: The overall response rate was 46%. The majority of respondents (92%) were aware of the relationship between antibiotic use and antibiotic resistance, and respondents estimated that 5% of their patients had URI symptoms at their office visits. Overall, 56% of respondents reported that they would prescribe an antibiotic for uncomplicated bronchitis and 43% for the common cold. OB/GYNs with the fewest years of experience were less likely than those with the most years of experience to report prescribing for uncomplicated bronchitis (Odds ratio (OR) 0.46, 95% confidence interval (CI) 0.23 to 0.91) or the common cold (OR 0.44, CI 0.22 to 0.89). The majority of respondents (60%) believed that most patients wanted an antibiotic for URI symptoms, with male OB/GYNs being more likely than female OB/GYNs (OR 2.1, CI 1.2 to 3.8) to hold this belief. Both male OB/GYNs (OR 1.9, CI 1.1 to 3.4) and rural practitioners (OR 2.1, CI 1.1 to 4.0) were more likely to believe that it was hard to withhold antibiotics for URI symptoms because other physicians prescribe antibiotics for these symptoms. OB/GYNs who believed that postgraduate training prepared them well for primary care management were more likely than those who did not (OR 2.1, CI 1.1 to 4.2) to believe that they could reduce antibiotic prescribing without reducing patient satisfaction. CONCLUSION: Multiple demographic factors affect attitudes and reported practices regarding antibiotic prescribing. However, in view of the low proportion of office visits for URIs, an OB/GYN-specific program is not warranted. PMID- 16040324 TI - Complexity of vaginal microflora as analyzed by PCR denaturing gradient gel electrophoresis in a patient with recurrent bacterial vaginosis. AB - OBJECTIVE: Gardnerella vaginalis has long been the most common pathogen associated with bacterial vaginosis (BV). We aimed to test our hypothesis that symptoms and signs of BV do not necessarily indicate colonization by this organism, and often will not respond to standard metronidazole or clindamycin treatment. METHODS: Using a relatively new molecular tool, PCR denaturing gradient gel electrophoresis (DGGE), the vaginal microflora of a woman with recalcitrant signs and symptoms of BV was investigated over a 6-week timeframe. RESULTS: The vagina was colonized by pathogenic enterobacteriaceae, staphylococci and Candida albicans. The detection of the yeast by PCR-DGGE is particularly novel and enhances the ability of this tool to examine the true nature of the vaginal microflora. The patient had not responded to antifungal treatment, antibiotic therapy targeted at anaerobic Gram-negative pathogens such as Gardnerella, nor daily oral probiotic intake of Lactobacillus rhamnosus GG. The failure to find the GG strain in the vagina indicated it did not reach the site, and the low counts of lactobacilli demonstrated that therapy with this probiotic did not appear to influence the vaginal flora. CONCLUSIONS: BV is not well understood in terms of its causative organisms, and further studies appear warranted using non-culture, molecular methods. Only when the identities of infecting organisms are confirmed can effective therapy be devized. Such therapy may include the use of probiotic lactobacilli, but only using strains which confer a benefit on the vagina of pre- and postmenopausal women. PMID- 16040325 TI - Diagnosis of bacterial vaginosis from self-obtained vaginal swabs. AB - OBJECTIVE: To determine the concordance between vaginal fluid Gram stains and pH obtained at speculum exam with similar stains and pH prepared from self-obtained vaginal swabs. METHODS: Using vaginal fluid Gram stain, 129 pregnant women were screened for bacterial vaginosis at 24 to 29 weeks' gestation. Two smears were collected from each woman during the same prenatal visit: the first was prepared from a self-obtained vaginal swab and the second from a physician-obtained speculum examination. Vaginal pH was recorded for each swab. Kappa coefficient was used to quantify agreement between the two sets of results. RESULTS: When compared with the physician-obtained smear, the ability of the self-obtained Gram stain to diagnose bacterial vaginosis had a sensitivity of 77%, specificity of 97%, positive predictive value of 71% and negative predictive value of 97%. There was substantial agreement (weighted kappa=0.82) between the two techniques in the ability to determine the grade of vaginal flora. CONCLUSION: When compared with physician-obtained vaginal smears, self-obtained smears have substantial agreement in the diagnosis of bacterial vaginosis. PMID- 16040326 TI - Vaginal Candida parapsilosis: pathogen or bystander? AB - OBJECTIVE: Candida parapsilosis is an infrequent isolate on vaginal cultures; its role as a vaginal pathogen remains unstudied. This retrospective study of women with positive culture for C. parapsilosis sought to characterize the significance of this finding and its response to antifungal therapy. METHODS: From February 2001 to August 2002, we identified all individuals with positive fungal isolates among a population of women with chronic vulvovaginal symptoms. Charts of women with C. parapsilosis cultures were reviewed with regard to patient demographics, clinical presentation and therapeutic response. Mycological cure, defined as a negative fungal culture at the next office visit, and clinical cure, i.e. symptom resolution, were determined for each subject. RESULTS: A total of 582 women had positive vaginal cultures for 635 isolates, of which 54 (8.5%) were C. parapsilosis. The charts of 51 subjects with C. parapsilosis were available for review and follow-up cultures and clinical information were available for 39 (76.5%). Microscopy was positive in 9 (17.6%). Antifungal treatment resulted in mycological cure in 17/19 patients with fluconazole, 7/7 with butoconazole, 6/6 with boric acid, 1/1 with miconazole and occurred spontaneously in 6/7: 24/37 (64.9%) patients with a mycological cure experienced clinical cure. CONCLUSIONS: Although C. parapsilosis is often a cause of vaginal symptoms, it seems to respond to a variety of antifungal agents and may even be a transient vaginal colonizer. PMID- 16040327 TI - Evaluation of relative yeast cell surface hydrophobicity measured by flow cytometry. AB - OBJECTIVE: To develop an efficient method for evaluating cell surface hydrophobicity and to apply the method to demonstrate the effects of fungal growth conditions on cell surface properties. METHODS: Yeast isolates were suspended in phosphate-buffered saline and mixed with deep blue-dyed polystyrene microspheres. Flow cytometry was used to detect the degree of microsphere binding to yeast cells. Different strains of yeast were compared for intrinsic microsphere binding activity and changes in growth conditions were invoked to modify the relative surface hydrophobicity. RESULTS: Commercially available blue dyed polystyrene microspheres showed strong fluorescence in the FL3 channel, whereas yeast cells did not show appreciable FL3 fluorescence. Microspheres and yeast were generally distinguishable on the basis of size revealed by forward light scatter. This method showed a wide variation in intrinsic cell surface hydrophobicity among Candida albicans strains. Likewise, variation in hydrophobicity of non-albicans yeast species was observed. Growth on solid media, incubation at 25 degrees C, or 250 mg/dl glucose concentration increased hydrophobicity compared with growth in liquid media, incubation at 37 degrees C, or 50 mg/dl glucose, respectively. Growth in 1 x 10(-9) M estradiol had no appreciable effect on hydrophobicity. CONCLUSIONS: Stained latex microspheres fluoresced in the FL3 channel of the flow cytometer and bound to yeast cells to an extent related to the surface hydrophobicity of the yeast. Binding detected by flow cytometry showed that clinical yeast isolates varied in intrinsic binding capacity and this binding ability was altered by different growth conditions. The implications for virulence regulation among yeast isolates are discussed. PMID- 16040328 TI - Acute placental infection due to Klebsiella pneumoniae: report of a unique case. AB - A 40-year-old woman, gravida 9, with seven healthy children and a history of one abortion (p 7 + 1), presented at 18 weeks of gestation with fever and malodorous vaginal discharge. Ultrasound revealed a macerated fetus. The placenta showed acute chorioamnionitis and acute villitis with microabscess formation. Blood and vaginal cultures both grew Klebsiella pneumoniae. This is the first reported case in English literature of Klebsiella pneumoniae causing suppurative placentitis leading to fetal demise. PMID- 16040329 TI - Actinomyces associated with persistent vaginal granulation tissue. AB - BACKGROUND: We report a case of symptomatic actinomycosis associated with vaginal suture erosion and granulation tissue refractory to conservative management, in an outpatient setting. CASE: Three months after total vaginal hysterectomy and uterosacral ligament vaginal vault suspension, a woman complained of painless, intermittent vaginal discharge and spotting. Despite cauterization of granulation tissue, vaginal spotting persisted for another month. On re-examination, braided polyester suture that was found underlying the granulation tissue was removed. Recurrent symptoms, together with a biopsy revealing actinomycetes, prompted a trial of oral penicillin VK. With persistent symptoms and discomfort during attempts in the outpatient clinic, the woman eventually required suture removal in the operating room. Her symptoms subsequently resolved without recurrence, and no further antibiotic treatment was required. CONCLUSIONS: Actinomyces may be associated with persistent granulation tissue and vault suspension suture material. In rare circumstances, when tissue debridement and suture removal in the clinic is unsatisfactory, surgical intervention in the operating room may be necessary. Ten days of antibiotic therapy alone did not eradicate the granulation tissue, and symptoms resolved only after complete removal of the underlying permanent suture. PMID- 16040330 TI - New developments in the pathogenesis of systemic sclerosis. AB - Systemic sclerosis (SSc) is characterized by extensive fibrosis, vasculopathy and activation of the immune system. Fibrosis can be caused by profibrotic cytokines, such as transforming growth factor-beta (TGFbeta), interleukin-4 (IL-4), platelet derived growth factor (PDGF), and connective tissue growth factor. Vasculopathy can be caused by TGFbeta, PDGF, while paucity of vessels in skin lesions can be attributed to anti-endothelial cell autoantibodies. Recent studies have suggested that the activation of the immune system is of paramount importance in the pathogenesis of SSc. T Cells are activated by antigen, infiltrate early the skin lesions in SSc, and produce the profibrotic cytokine IL-4. They are also required for autoantibody production. B cells may contribute to fibrosis, as deficiency of CD19, a B cell transduction molecule, results in decreased fibrosis in animal models of fibrosis. These new developments have direct impact on the treatment of SSc. Medications directed against immune cells or harmful soluble factors in small trials in SSc are encouraging. PMID- 16040331 TI - Pemphigus following hepatitis B vaccination--coincidence or causality? AB - Pemphigus is an autoimmune blistering disease caused by autoantibodies against epithelial intercellular components. Its etiology is unknown, and neoplasms, antecedent infections or medications are considered possible triggering factors for the disease in some cases. We describe the first case of pemphigus following a hepatitis B virus vaccination. We suggest that in some cases vaccination may be the triggering factor for pemphigus in genetically predisposed individuals and physicians should be aware of this possible association. PMID- 16040332 TI - Cytomegalovirus infection in central European vitiligo patients? PMID- 16040333 TI - IVIg therapy in autoimmunity and related disorders: our experience with a large cohort of patients. AB - Intravenous immunoglobulin (IVIg) is used to treat a number of immune deficiencies and autoimmune diseases. It has been shown that IVIg contains anti idiotypic antibodies, which explains its immunomodulatory action. In murine models, recent investigations have demonstrated that IVIg can prevent and reduce the affliction by systemic lupus erythematosus (SLE), antiphospholipid syndrome (APS) and scleroderma. Relevant disease-specific fractions of IVIg were able to reproduce and even enhance the therapeutic effect in a murine model.IVIg treatment before tumor resection in rodents inoculated with melanoma and sarcoma cells dramatically improved the cure rate (50%) in comparison to the control group (0%). In patients affected by SLE, several clinical manifestations responded to IVIg treatment including serositis, hematological manifestations, treatment-resistant nephritis and central nervous system involvement. Similarly, in women with recurrent fetal loss due to APS, IVIg was able to diminish the abortion rate. Vasculitides such as Churg-Strauss' and Wegener's and skin fibrosis in patients affected by scleroderma improved after IVIg treatment. In agreement with in vitro investigations, prolonged survival has been noted in cancer patients treated with IVIg. We suggest that in the presence of a steroid and immunosuppressive-resistant autoimmune disease, IVIg is a rational and safe choice. PMID- 16040334 TI - Monozygotic twins discordant for epilepsy differ in the levels of potentially pathogenic autoantibodies and cytokines. AB - Can autoantibodies (Ab's) and cytokines play a role in epilepsy?Monozygotic twins discordant for epilepsy (most probably Rasmussen's encephalitis (RE)), compared to 49 neurologically intact controls, were both found to contain in their serum (at the time of epilepsy diagnosis) significantly elevated levels of specific Ab's against peptide B (amino acids 372-395) of the ionotropic glutamate receptor of AMPA subtype 3 (i.e. GluR3B peptide). Interestingly, both twins also had clinically elevated levels of Ab's to double-stranded (ds) DNA, glutamic acid decarboxylase, nuclear antigens, beta2-glycoprotein I and cardiolipin, as in "classical" autoimmune diseases. Both twins also had significantly elevated levels of IFNgamma, TNFalpha, IL-4 and IL-10 in the serum, compared to the controls. Comparing the twins revealed that the epileptic twin had significantly higher levels of five of the above anti-self Ab's, but significantly lower levels of all four cytokines compared to her healthy sister. Importantly, the epileptic twin, alike three other RE patients tested herein, contained elevated levels of Ab's to GluR3B and dsDNA also in cerebrospinal fluid (CSF) (unavailable of the healthy twin). Our results suggest that the various autoimmune Ab's studied herein, all of which are known already to have a potential to be pathogenic in the nervous system and/or peripheral organs, may play a role in some types of epilepsy. The titer of such Ab's and of key cytokines may be crucial for either facilitating or arresting the development of epilepsy. Our findings also show that anti-GluR3B Ab's in serum are not necessarily detrimental (their presence in the CSF may be more dangerous), and that they are not a mere side effect of already existing epilepsy, as they were found herein in serum of a healthy individual. These findings and suggestions may be of clinical importance and call for further studies. PMID- 16040335 TI - Association of a CTLA-4 3' untranslated region (CT60) single nucleotide polymorphism with autoimmune thyroid disease in the Japanese population. AB - The etiology of the autoimmune thyroid diseases (AITDs), Graves' disease (GD) and Hashimoto's thyroiditis is largely unknown. However, genetic susceptibility is believed to play a major role. The cytotoxic T-lymphocyte antigen-4 (CTLA-4) gene, encoding a negative regulator of the T-lymphocyte immune response, had been reported to be associated and/or linked to AITD. Recently, AITD susceptibility in the Caucasians was mapped to the 6.1-kb 3'UTR of the CTLA-4 gene, in which the single-nucleotide polymorphism (SNP), CT60, was most strongly associated with AITD. In order to determine the association of the CTLA-4 gene with AITD in the Japanese, case-control association analysis for the CT60 of the CTLA-4 gene using 264 AITD patients and 179 healthy controls was done. The frequency of the disease susceptible G allele of the CT60 of the Japanese control was higher than that of the Caucasians (72.6 vs. 52.3%). However, the G allele of the CT60 was associated with GD (84.0 vs. 72.6%, P=0.0008) and AITD (80.1 vs. 72.6%, P=0.009) in the Japanese. Furthermore, the G allele of the CT60 was associated with the increased risk for GD [P=0.004, odds ratio (OR)=2.0] and AITD (P=0.03,OR=1.6) in a recessive model. These results suggested that the CTLA-4 gene is involved in the susceptibility for GD and AITD in the Japanese. PMID- 16040336 TI - Autoantibodies profile of rheumatoid arthritis patients during treatment with infliximab. AB - PURPOSE: Therapy with TNFa blocking agents has been associated with increased rate of anti-nuclear antibodies (ANA) and rare cases of lupus like syndromes. Our aim was to prospectively analyze a wide array of autoantibodies in rheumatoid arthritis (RA) patients before and 14 weeks after starting infliximab. MATERIAL AND METHODS: In this study, 26 consecutive active RA patients participated. All treated with infliximab at a dosage of 3 mg/kg on week 0, 2, 6 and every 8 weeks, along with weekly low dose methotrexate. Patients were evaluated at week 0 and 14. Clinical assessment included the number of tender and swollen joints, duration of morning stiffness, adverse events (AE) (including SLE-like) and ESR. Sera were collected before the 1st infusion of infliximab at week 0 and 14. The autoantibodies studied were: fluorescent ANA, anti-double-stranded-DNA (anti-ds DNA), IgG and IgM anti-cardiolipin (ACA), anti-histone- H1 and C (H1, H2A, H2B, H3, H4), anti-SSA, -SSB, -ENA, -scleroderma 70, -thyroid peroxidase (TPO) and neutrophilic cytoplasmatic (ANCA) antibodies. RESULTS: Of 26 patients, 17 were women. A significant decrease in duration of morning stiffness, number of tender and swollen joints and ESR were observed between week 0 and 14. During follow up (mean of 20.5+/-7.3 months), 9 patients stopped infliximab due to inefficacy or AE (most of them after the 4th infusion). Two patients developed lupus-like phenomena. ANA was found positive at baseline in 7 out of 26 patients. In 5 of them, an increase in the titer of ANA was observed at week 14. ANA negative turned positive for 8 patients. A significant increase of anti-cardiolipin (ACA) IgM levels was observed in 8 patients and of ACA-IgG in 6, in parallel with ANA seroconversion. The mean level of anti-double-stranded-DNA (anti-ds-DNA) -IgG significantly increased from 66+/-33 to 93+/-68 IU/ml, in 4 patients to pathological levels. Four patients demonstrated an increase in anti-histone H1. Levels of ANCA, anti-ENA, -SSA, -SSB, -RNP, -scleroderma70 and -thyroid peroxidase (TPO) were negative in all patients and remained unchanged during the study. Cessation of treatment with infliximab was found to be associated with the appearance of ANA. CONCLUSION: An increased titer or a new appearance of ANA was observed in 12 out of 26 patients. The main autoantibodies found were anti-ds DNA, ACA-IgM and -IgG and anti-histone. In our cohort, the appearance of some autoantibodies seemed to predict late cessation of treatment. PMID- 16040337 TI - Suppression by mAbs against DQB1 peptides of in vitro proliferation of AChR specific T cells from myasthenia gravis patients. AB - It has been indicated that multiple genes, including HLA genes, are collectively involved in the susceptibility to myasthenia gravis (MG). DQB1 alleles represent one of those associated with MG. We have prepared B-cell hybridomas that produce mAbs against peptides corresponding to the tip of the MHC antigen-binding cavity (region 70-90) of alleles DQB1*02, *03, *05 and *06. The mAbs bound to DQ molecules isolated from cells. In the assays using peripheral blood lymphocytes (PBL) from patients with MG, the mAbs against peptides of the correlate HLA DQ sequences inhibited the in vitro proliferation of acetylcholine receptor (AChR) specific T cells. The results indicate that the function of disease-related MHC alleles may be blocked by directly and selectively targeting the antigen presenting region on these MHC molecules. The results also suggest that DQ molecules are one of those involved in the restriction of autoimmune anti-AChR responses in MG. The strategy could provide an effective means for immunointervention in MG. It may also potentially be adapted for down-regulation of undesirable immune responses such as in other autoimmune diseases, allergic reactions, or clinical conditions where immune responses to a therapeutic protein develop. PMID- 16040338 TI - Atheroprotective immunization with MDA-modified apo B-100 peptide sequences is associated with activation of Th2 specific antibody expression. AB - OBJECTIVE: The objective of this study was to evaluate if immunization with MDA modified human apo B-100 fragments is associated with a shift in the Th1/Th2 balance. METHODS AND RESULTS: Apo E deficient mice were immunized with one of the peptides (P45; amino acids 688-707, P74; amino acids 1123-1142 or P240; amino acids 3613-3632) at 6, 9 and 11 weeks of age and compared to controls given carrier alone. Immunization with P45 and P74 reduced atherosclerosis in the aorta of 25-week-old mice by 48% (p=0.02) and 31% (p=0.06) and macrophage content in atherosclerotic plaques by 33% (p=0.02) and 39% (p=0.02), respectively. The levels of Th2-specific IgG1 against each peptide increased more than 50-fold in response to immunization, whereas the levels of specific IgM and Th1-associated IgG2a were only marginally affected. However, there was an increase in the plaque expression of both Th1 and Th2 cytokines as assessed by real time PCR. Immunization with P240, a non-homologous peptide used as control, induced a 10 fold increase of specific IgG1 but did not influence atherosclerosis or plaque content. CONCLUSIONS: Immunization with MDA apo B-100 fragments induce a shift from Th1 to a Th2 specific oxidized LDL antibody expression, but without a concomitant downregulation of plaque IFN-gamma expression. PMID- 16040339 TI - Concurrent HLA-related response factors mediate recombinant hepatitis B vaccine major adverse events. AB - Recombinant hepatitis B (HB) vaccines have successfully reduced infection, cirrhosis and carcinoma, but questions have endured about causality of serious adverse events following vaccination. After an event in a pediatric patient an investigation reviewed HLA vaccine response effects and analyzed genetics in reported cases. There are apparent common causal immune mechanisms among reported adverse events. HLA class II alleles/haplotypes linked to HB vaccine cellular/non response and Crohn's disease can create conditions that actively/passively amplify, respectively, all or other components of the immune response to the HB vaccine. Presence of the HLA class I allele A2 can result in heavy cytotoxic T cell activation and vaccine/self-peptide presentation to immune cells. If HLA autoimmune susceptibility alleles/haplotypes are present that control other immune response components, the probability is elevated that these will activate cross-reactive immune cells; the cells, their inflammatory secretions and/or auto antibodies may initiate adverse events reflecting those susceptibilities. Probable DRB1 amplifying alleles are noted. High-resolution DNA typing and results analysis are described to test the hypothesis in known HB vaccine adverse event patients. Possible practical applications stemming from hypothesis validation are described. PMID- 16040340 TI - Alternative transcripts of rat slc19a1: cloning, genomic organisation, tissue specific promoters and alternative splicing. AB - Recently, the rat genome project revealed the genomic sequence of slc19a1, coding for the methotrexate carrier-1, identical to the reduced folate carrier-1 of humans, on rat chromosome 20. At the same time, we have cloned and analysed the complete or partial cDNAs of now at least six different transcripts from rat liver and kidneys. Alignment with the genomic sequence revealed seven exons. The first two non-coding exons, exon I and Ia were used alternatively in kidneys and liver, respectively, suggesting usage of alternative promoters. Three minor mRNA forms resulted from absent splicing of intron III, a shortened exon III (exon IIIa), and a shortened exon IV (exon IVa). The minor transcripts were predicted to result in translation products with 7 or 6 instead of 12 transmembrane domains (TMDs) and a peptide mass of 38, 39 and 40 kDa instead of 58 kDa. PMID- 16040341 TI - Identification and characterization of a canine highly similar to retinoic acid receptor alpha. AB - A canine highly similar to retinoic acid receptor alpha (canine HS-RARa) cDNA was isolated from the spleen tissue. A database search and the alignment revealed that the canine cDNA was most similar to highly similar type of human RARa and was named canine HS-RARa. The expression of the genes encoding RARa in the dog was the highest in the testis and moderate in the blood, lymph node, mammary gland, pancreas, salivary gland, spleen, thyroid gland, tonsil and uterus. The nucleotide sequence encoded the 462-amino acid containing the conserved sequence motif of RARa. Though the amino acid sequences were well-conserved among species, some unique arrangements were observed within each class. In the phylogenetic analysis, each species separated according to their class. In the branch of mammals, the dog is in the cluster of humans, mice and western wild mice. However, hamsters and rats formed another branch. PMID- 16040342 TI - Molecular cloning and characterization of beta-defensin cDNA expressed in distal ileum of buffalo (Bubalus bubalis). AB - Defensins play a prominent role in protection of various epithelial surfaces. In this study, we have cloned and characterized the mRNA from the distal ileum of Bubalus bubalis. Total RNA after isolation from ileal epithelium was reverse transcribed to synthesize cDNA using primers designed by taking conserved region of cattle enteric beta-defensin (EBD) mRNA, goat beta-defensin 2 (BD 2) and cattle lingual antimicrobial peptide (LAP) mRNA sequences. The PCR amplified cDNA of 254 bp was ligated to pDrive cloning vector and transformed into XL-blue strain of E coli. The sequence analysis indicated 29 nucleotide substitutions with reported cattle EBD mRNA sequence sharing 86.2% homology, 92.1% with cattle LAP, 81.6% with cattle tracheal antimicrobial peptide and 84.6% with goat BD 2. The deduced amino acid sequence encodes for a 64 amino acid precursor peptide. Both nucleotide and amino acid sequence homology shows that the cloned sequence is closer to cattle LAP. PMID- 16040343 TI - Cloning and characterization of a ribonuclease L inhibitor from the silkworm, Bombyx mori. AB - The ribonuclease L (RNase L) pathway plays an important role in the response of cells to double-stranded RNA (dsRNA) during the events such as virus infection. Ribonuclease L inhibitor (RLI) belonging to the ABC transporter family is known as a regulator of the RNase L pathway. The homologs of RLI were reported in many organisms including the fruit fly and mosquito, but their functions in insects and arthropods have not been elucidated to date. In the present study, we cloned a cDNA of a silkworm RLI homolog, termed BmRLI, and its nucleotide sequence was determined. RT-PCR analysis revealed that the expression of BmRLI mRNA was marked in the testis, ovary and fat body. From the cDNA, recombinant protein with an apparent molecular mass of 69 kDa was expressed in Escherichia coli and cultured insect cells. Although no obvious effect of up-regulation of the BmRLI expression on RNAi was observed, its down-regulation slightly reduced RNAi efficiency. PMID- 16040344 TI - A novel ABA-dependent dehydrin ERD10 gene from Brassica napus. AB - A new dehydrin ERD10 gene was cloned and characterized from Brassica napus (designated as Bndhn ERD10). The full-length cDNA of Bndhn ERD10 was 1114 bp and contained an open reading frame of 816 bp encoding a protein of 271 amino acid residues. The deduced Bndhn ERD10 protein contained an 8-serine residue domain and two conserved repeats of the characterized lysine-rich-K-segment (KIKEKLPG). Analysis of full-length cDNA and genomic DNA indicated that there were no introns in Bndhn ERD10 gene. The promoter of Bndhn ERD10 was further obtained by genomic walking technology, and analysis of the promoter indicated that the regulation of Bndhn ERD10 was ABA-dependent. Semi-quantitative RT-PCR of different tissues in unstressed B. napus plants indicated that the transcript of Bndhn ERD10 was more abundant in leaf than in stem and root. The expression profiles of Bndhn ERD10 in B. napus seedlings under various stress conditions including cold, salt and ABA were also investigated. Upon cold, salt and ABA stresses, increased transcript accumulations of the Bndhn ERD10 mRNAs were detected in young leaves 8 h after treatment. PMID- 16040345 TI - Analysis and characterization of the transcriptional unit of a new Mytilus galloprovincialis (Mollusca: Bivalvia) hsp70 gene. AB - We isolated, analysed and characterized the transcriptional unit of an hsp70 gene of Mytilus galloprovincialis (Mollusca: Bivalvia). Sequence analysis of a total of 3653 bp, revealed a 1914 bp ORF encoding for a 637-amino acid polypeptide that exhibits all the characteristic signatures of the cytoplasmic members of the HSP70 family. The ORF is not interrupted by introns and its 5'-flanking region contains five putative heat shock elements (HSEs), both indicating for an inducible hsp70 gene. Furthermore, nucleotide and amino acid sequence alignments with HSP70 genes from several molluscan and non-molluscan species confirmed that the newly isolated gene, the first complete in the genus Mytilus and in the Mollusca phylum, is a cytoplasmic and inducible hsp70 gene. PMID- 16040346 TI - Specific amplification of cDNA ends (SPACE): a new tool for the analysis of rare transcripts and its application for the promoter analysis of killer cell receptor genes. AB - The expression control of activating and inhibitory killer cell Ig-like receptors (KIR) on natural killer (NK) cells is highly relevant for the initiation of NK cell mediated cytolysis and cytokine secretion. Transcription start points of nine human KIR genes from two Caucasian donors and the NK cell line NK3.3 were investigated. To overcome sensitivity problems due to the low abundance of the respective transcripts, a novel protocol, specific amplification of cDNA ends (SPACE) with superior specificity and sensitivity was applied. A total of 235 individual SPACE clones resulting from different KIR genes were analysed and revealed a series of transcription start sites tightly clustered between 10 and 60 bp upstream of the start codon. The comparison of the adjacent putative promoter region of the human, chimpanzee and macaque KIR genes revealed a very high conservation for almost all of the KIR family members. An inter-gene and inter-species comparative approach revealed transcription factor binding sites at regions of maximal homology for all primate KIR genes analysed. PMID- 16040347 TI - Analysis of the 3' UTR of the ART3 and ART4 gene by 3' inverse RACE-PCR. AB - 3' Rapid amplification of cDNA ends (3' RACE) is a polymerase chain reaction (PCR) based technique which has been developed to analyse 3' ends of partially known cDNA sequences. To improve the effectiveness of the technique, many investigators have modified the RACE protocol. Here, we describe an alternative procedure for analysing 3' mRNA ends which is based on DNA ligase-mediated self circularization and inverse PCR. This technique is simple and characterized by the exclusive use of gene-specific primers and the absence of unspecific adaptor sequences to obtain highly specific PCR products. We applied the method to analyze the 3' UTR of human mono-ADP-ribosyltransferase (ART) 3 mRNA in testis and heart muscle and of ART4 mRNA in HEL cells. The obtained sequences of ART3 and ART4 mRNA corresponded to data base entries of the respective mRNAs. No adenylate/uridylate-rich elements (AREs) were found in the 3' UTR of ART3 mRNA while one ARE class I motif was detected in the 3' UTR of ART4 mRNA. PMID- 16040348 TI - Sequence analysis of canine and equine ferritin H and L subunit cDNAs. AB - Canine and equine ferritin H and L subunit cDNA clones were obtained using reverse transcriptase-polymerase chain reaction (RT-PCR) and TA cloning from various tissues. Canine liver and spleen ferritin H subunit cDNA clones contained an open reading frame for the same 182-amino acid protein as that reported in canine brain ferritin H subunit cDNA although there were substitutions in the 3' noncoding regions. Ferritin L subunit cDNA clones from canine liver, spleen, and kidney showed identical coding sequences encoding the 174-amino acid protein except for a single nucleotide substitution in kidney (C474G). The H subunit nucleotide sequences of equine leukocyte and spleen were identical to the fragment encoding the 181-amino acid protein in equine peripheral blood mononuclear cells, with the exception of one substitution seen in both leukocyte and spleen sequences (C234T). The nucleotide sequence of equine leukocyte ferritin L subunit showed 7 substitutions compared with the published equine liver L subunit sequence with two substitutions at positions 281 and 282 resulting in an amino acid substitution of P94L. The amino acid residues involved in the ferroxidase center and in iron nucleation were perfectly conserved in H and L subunits of canine and equine ferritins, respectively. PMID- 16040349 TI - Cloning of rat p47phox and comparison with human p47phox. AB - A cDNA encoding rat p47phox was cloned from rat spleen cDNA library, utilizing rapid amplification of cDNA ends. The open reading frame corresponded to 389 amino acids: It contained the phagocyte oxidase homology domain, two Src homology 3 domains and a proline rich region, all of which are conserved in mammalian p47phox sequences. Rat p47phox displayed the highest degree of identity to mouse p47phox (94%). We expressed and purified rat p47phox as a glutathione S transferase fusion protein, and found that the rat protein could replace human p47phox in a cell-free activation system for human NADPH oxidase, giving about half activity. Although rat 12-lipoxygenase interacted with human p47phox in a yeast two-hybrid system, this was not the case for rat p47phox. PMID- 16040350 TI - Cloning and characterization of the rpoE gene encoding an RNA polymerase sigmaE factor from the deep-sea piezophilic Shewanella violacea strain DSS12. AB - The rpoE gene encoding an RNA polymerase sigmaE subunit was isolated from a gamma phage library of the deep-sea piezophilic and psychrophilic bacterium Shewanella violacea strain DSS12. Structual analysis showed that the gene organization of the fragment containing S. violacearpoE was the l-aspartate oxidase-coding gene, rpoE, rseA, rseB and rseC in that order, the same as in the case of Photobacterium profundum SS9 and Escherichia coli K-12. The cloned gene, 576 bp in length, was found to encode a protein consisting of 192 amino acid residues with a molecular mass of 21,806 Da. Amino acid alignment of the RpoE protein showed that the functional domains responsible for DNA recognition, DNA melting, core binding, and RseA interaction were highly conserved. We purified hexahistidine-fused RpoE protein by constructing an overexpression plasmid. Core binding analysis revealed that the cloned RpoE protein has the ability to bind with core RNA polymerase as a sigma factor. PMID- 16040351 TI - Identification and characterization of IS1112 and IS1113 insertion element sequences in Xanthomonas oryzae pv. oryzae. AB - Two new insertion sequences (IS1112 and IS1113) were identified in the genome of Xanthomonas oryzae pv. oryzae, the causal agent of bacterial blight of rice. Three copies of IS1112 were trapped, one containing 1052-bp and the other two with 1055-bp. They all have 25-bp imperfect inverted repeats with a 3-bp duplication at the site of insertion. They contain an open reading frame (ORF) of 317 and 318 amino acid residues, respectively. IS1113 is 1306-bp, contains 25-bp imperfect terminal inverted repeats, and is flanked by a 9-bp direct repeat at the site of insertion. It contains an ORF of 395 amino acid residues. PMID- 16040352 TI - A fifth member of the tomato 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase gene family harbours a leucine zipper and is anaerobically induced. AB - Using the leucine zipper domain of a small anaerobically induced bZIP transcription factor in a yeast two hybrid screen, anaerobically induced genes were identified. One peptide corresponds to an anaerobically induced IDS4-like protein that maybe involved in G-protein signaling. Surprisingly, another interacting peptide corresponds to a novel anaerobically induced 1 aminocyclopropane-1-carboxylic acid (ACC) oxidase, designated ACO5. ACO5 harbours a leucine zipper and transcription is mainly induced in fruits and to a lesser extend in leaves. The role of ACO5 in the low oxygen response of tomato is discussed. PMID- 16040353 TI - Qi-training (qigong) enhanced immune functions: what is the underlying mechanism? AB - The authors observed that Qi-training enhances immune function and modulates neurohormone concentrations. The exact signal and priming mechanism for enhanced neutrophil function by Qi-training has not yet been demonstrated. This study investigated the effect of Qi-training on intracellular signaling leading to the enhancement of immune function. The growth hormone (GH) concentrations and O2- production by neutrophils (PMNs) was significantly increased after 1 h of Qi training compared with the basal state. To verify that endogenous GH mediates the priming of PMNs, serum obtained from elderly subjects in the basal state and after Qi-training was incubated with neutrophils isolated from young subjects for 60 min and triggered with N-formyl-1-methionyl-1-leucyl-1-phenylalanine (fMLP). Significant O2- production was observed in the PMNs incubated with serum collected after a Qi-training (p < .05). On the other hand, the priming effect on the PMNs was abolished in Qi-training sera depleted of endogenous GH with anti human GH polyclonal antibody (p < .01) and the tyrosine kinase inhibitor, genistein (p < .01). The authors suggest that the endogenous GH released during and immediately after Qi-training mediates the priming events through tyrosine kinase activation in PMNs. PMID- 16040354 TI - Projected color slides as a method for mass screening test for color vision deficiency (a preliminary study). AB - This article compared the efficiency of the mass screening test with projected color slides in detecting color-blindness with the authentic classic method of Ishihara. The study was conducted in a randomly selected lecture room with 104 students aged between 19-25 years (median 21). Using Ishihara projected slides, performed mass screening test. Re-testing was done individually with printed Ishihara plates. Six male and one female with color-blindness were detected. The frequency of color-blindness was 13.6% among males, with a total incidence of 6.7%. The results of two testing methods were compared statistically. Sensitivities and specificities of both tests were 100%. Using projected slides of Ishihara plates instead of the authentic method is an effective and timesaving method for detecting color-blindness. This method can be suggested as a mass screening test and might be beneficial in detecting color-blindness in large populations such as students, soldiers, and so on. PMID- 16040355 TI - Age dependent penetrance of three different superoxide dismutase 1 (sod 1) mutations. AB - The age of onset of motor neuron disease in Cu/Zn superoxide dismutase 1 (SOD1) mutation carriers are variable, commencing at any time from the second decade. The authors performed a retrospective analysis of family information in pedigrees dating back to the 1780s, to determine the age-dependent penetrance of three different SOD1 mutations: Glu100Gly, Ile113Thr, and Val148G1y. The penetrance of symptomatic MND in these three SOD1 mutations was greater than 95% by the age of 78. The affected family members with the Val148Gly mutation had the worst prognosis, with a mean age of death of 46.1 years, compared to 54.2 years for the Glu100Gly mutation and 59.9 years for Ile113Thr mutation. Kaplan-Meier survival curves showed that survival of the 3 SOD1 mutation families, when combined, was reduced by nearly 10 years with the mean age of death for all SOD1 mutation carriers being 52.6 years compared to 62.5 years for the control individuals. The SOD1 mutation group also resulted in earlier death compared to sporadic MND, which from natural history studies is 61.4 years. This may reflect that the SOD1 mutation is associated with more progressive and rapid disease, as the age of onset of disease was not earlier. This information would have important implications for genetic counseling of members of individual SOD1 mutation carrier families. PMID- 16040356 TI - Neuropsychological measures and single photon emission computed tomography in the differentiation and classification of cerebral perfusion deficits in Alzheimer's dementia. AB - The accurate diagnosis of Dementia of the Alzheimer's Type (DAT) continues to be an area of difficulty for the fields of neuropsychology and neurology. The introduction of new medications that appear to mediate the insidious progression of the disorder increases the need for timely differentiation of DAT from other dementia-related disorders. The present study examined the relationship between hemispheric differences in regional cerebral blood flow with corresponding lateral neuropsychological processing deficits in patients with DAT. Eighty patients with a diagnosis of DAT were administered Single Photon Emission Computerized Tomography (SPECT) scans and a battery of left and right hemisphere neuropsychological-based tasks. The results of ANOVA indicated that patients with DAT who were not suffering from perfusion deficits exhibited significantly fewer neuropsychological deficits than did patients with DAT who had perfusion deficits. The neuropsychological tests that measured verbal ability, logical memory, word-pair learning, reading, arithmetic, and visual-perceptual organization were all significantly lower in the group with perfusion deficits. Further analysis indicated that patients with left hemisphere perfusion deficits tended to have poorer neuropsychological skills than did individuals with right hemisphere perfusion deficits, diffuse perfusion deficits, and no perfusion deficits. A Stepwise Discriminant Analysis was unable to use the neuropsychological variables to classify patients accurately into perfusion deficit groups. PMID- 16040357 TI - EFFECT of nicotine on hippocampal nicotinic acetylcholine alpha7 receptor and NMDA receptor subunits 2A and 2B expression in young and old rats. AB - Nicotinic acetylcholine (nAChR) and N-methyl-D-aspartate receptors (NMDARs) play critical roles in memory function. This study administered chronic nicotine to determine the alterations of N-methyl-D-aspartate receptor subunit 2A and 2B (NR2A, NR2B) and the alterations of alpha7nAChR receptor. It was determined that the effectivity of nicotine and the data support that nicotine increases hippocampal NR2A and B expression. Additionally, the role of nicotine in the cognitive improvement was not supported by the antioxidative mechanisms or the authors observed no effect of nicotine on lipid peroxidation at the hippocampus. PMID- 16040358 TI - The face, beauty, and symmetry: perceiving asymmetry in beautiful faces. AB - The relationship between bilateral facial symmetry and beauty remains to be clarified. Here, straight head-on photographs of "beautiful" faces from the collections of professional modeling agencies were selected. First, beauty ratings were obtained for these faces. Then, the authors created symmetrical left left and right-right composites of the beautiful faces and asked a new group of subjects to choose the most attractive pair member. "Same" responses were allowed. No difference between the left-left and right-right composites was revealed but significant differences were obtained between "same" and the left left or right-right. These results show that subjects detected asymmetry in beauty and suggest that very beautiful faces can be functionally asymmetrical. PMID- 16040359 TI - Effects of myofascial-meridian stimulation therapy (mmst) on shoulder pain. AB - Fifty patients with shoulder pain were classified into two groups according to the type of pain: a capsular-pattern group (N = 22) and a non-capsular-pattern group (N = 28). Both groups received myofascial-meridian stimulation therapy four times in two weeks. After therapy, the pain level was significantly lower in the capsular-pattern than in the non-capsular-pattern group. The capsular-pattern group showed a nonsignificant trend toward greater range of motion than the non capsular-pattern group. These data suggest that myofascial-meridian stimulation therapy might be useful as a new treatment to reduce pain and increase range of motion in patients with shoulder pain. PMID- 16040360 TI - Association analysis of serotonin transporter promoter gene polymorphism with ADHD and related symptomatology. AB - Attention deficit hyperactivity disorder (ADHD) is one of the most common childhood behavioral disorders. Genetic factors contribute to the underlying liability to develop attention deficit hyperactivity disorder. Several investigations have reported associations between ADHD and serotonin transporter promoter polymorphisms, but the results have been inconsistent. The present study did not find significant association between ADHD and serotonin transporter promoter polymorphisms, but did find an effect of serotonin transporter promoter polymorphisms on some ADHD symptomatology. Patients homozygous for the short allele showed more Withdrawn or Somatic complaint scores than subjects with the long allele. PMID- 16040361 TI - Neuromuscular responses of trained ballet dancers to postural perturbations. AB - The balance of trained ballet dancers and non-dancer controls was mechanically perturbed in order to evaluate the time of onset of muscle activation and the consistency of muscle activation. Results supported the prediction that ballet dancers have significantly faster long-latency (LL) neuromuscular responses than controls and are significantly more consistent in muscle activation. These findings indicate a superior postural control mechanism in trained dancers and may explain the ability of dancers to maintain static balances over a small base of support. PMID- 16040362 TI - Depression of h-reflex following carbonic anhydrase inhibition appears unrelated to changes in synaptic effectiveness. AB - Presynaptic inhibition (PI) of Ia afferents was examined as a possible contributor to the depression of the soleus H-reflex following carbonic anhydrase (CA) inhibition with Acetazolamide (ACZ). Ten males (aged 22-32) were studied in two randomized conditions, control and ACZ administration (250 mg 14, 8, and 2 h before testing) separated by at least one week. PI of soleus Ia afferents was indirectly assessed two ways: a conditioning stimulus of Ia afferents in the common peroneal nerve (N = 6), and heteronymous Ia facilitation from the quadriceps to soleus muscle (N = 4). Conditioning (C) of the soleus H-reflex (common peroneal nerve stimulation protocol) resulted in depression of the H reflex in the supine and standing position compared to the test (T, unconditioned) H-reflex in the same position. This result was unaltered following ACZ treatment. C (heteronymous facilitation protocol) resulted in facilitation of the H-reflex in the supine, but not the standing position. This result was unaltered following ACZ treatment. It was concluded that the depression of the H reflex following CA inhibition (present study; Brechue et al., 1997) appears to be unrelated to changes in the tonic level of PI of Ia afferents. The best hypothesis for the reduction in the H-reflex appears to be conduction block of the primary afferent fibers secondary to local increases in PCO2. PMID- 16040363 TI - Magnetic resonance imaging findings in pre-clinical Creutzfeldt-Jakob disease. AB - The diagnosis of probable Creutzfeldt-Jakob disease (CJD) requires compatible clinical manifestations, typical electroencephalographical findings, and 14.3.3 protein positive in cerebrospinal fluid. Actually, MRI findings are not necessary, but they may support this diagnosis. The authors report a patient with definitive diagnosis of CJD who showed in a first MRI study performed two years before the clinical onset of the disease the same hyperintensities in caudate nuclei that were found in the last MRI. The authors think that these findings could be useful in detecting asymptomatic patients who need more extensive study and following. PMID- 16040364 TI - Hemispheric integrative therapy in Landau-Kleffner syndrome: applications for rehabilitation sciences. AB - A case study is presented of a 14-year-old right-handed Caucasian female diagnosed with the Landau-Kleffner Syndrome (LKS) at the age 3 1/2 years. Her LKS symptoms presented with abrupt disruption in language after normal development, electroencephalogram (EEG) brain-wave abnormality, seizure activity, inability to read, and impairment in her motor skills. After 11 years of pharmacological and special education interventions with no significant improvement in any measurable area of function, a multimodal approach using techniques purportedly aimed at facilitating inter-hemispheric communication was provided. At completion of the program, EEG was controlled, reading, language, and auditory processing improved and objective behavioral-social measures improved significantly. PMID- 16040365 TI - Ketamine reduces lidocaine-induced seizures in mice. AB - Systemic toxic reactions to local anesthetics are brought about by absolute overdosage, and, most commonly, inadvertent intravascular injections. The anti convulsant action of ketamine has been studied. However, the effect of ketamine on lidocaine-induced convulsions has not been reported. This study investigated the effect of ketamine on lidocaine-induced seizures in mice. Mice (32-41 g) were divided into 2 groups, 15 in each group, and were pretreated with intraperitoneal normal saline solution or intraperitenoeal (ip) ketamine before lidocaine. Group 1 (N = 15) received 75 mg kg ip lidocaine; Group 2 (N = 15) received 20 mg kg ketamin ip; 5 min later 75 mg kg lidokaine ip were applied. Clinical features, incidences, latencies, durations, and mortality rate of convulsions were recorded. After 75 mg kg lidocaine injection, ataxia, loss of righting reflex, and generalized tonic-clonic convulsions were seen within 2-5 min in Group 1. Generalized tonic-clonic convulsions were seen in 8 mice and deep sedation was seen in 7 mice in Group 2 (p < .05). Generalized status epilepticus occurred in one mouse in both groups. Three mice from Group l and one mouse from Group 2 died during convulsions. There were no differences between the two groups with regard to the onset and duration of seizures (p > .05). It was concluded that ketamine significantly prevented lidocaine-induced generalized tonic-clonic seizures; on the other hand, the lethality of lidocaine was least reduced by ketamine. PMID- 16040366 TI - Spreading interviews over time in health surveys: do temporal variations of self reported alcohol consumption affect measurement? AB - OBJECTIVE: To address systematic variations related to the day of the interview in self-reports of alcohol consumption in telephone health surveys. The investigations include temporal clustering effects, prediction of alcohol consumption and variations across days by characteristics of respondents and interviewing period, and sensitivity to variations of measurements instruments. METHOD: Data at baseline collected in Spring 1999 from 2846 participants in a longitudinal probabilistic general-population survey in Switzerland were used. The study is representative for drinkers in Switzerland. Alcohol consumption measures include a 6-month quantity frequency and a 1-week graduated frequency measure. RESULTS: Evidence for systematic variations in self-reports related to the day of interview was found on the graduated-frequency measure even after controlling for sample characteristics. Similar variations on the quantity frequency measure were found, but were no longer significant after statistical control of the sample characteristics. CONCLUSIONS: Statistical inference in alcohol survey research by telephone interviews based on graduated-frequency measures with short reference period may be plagued with errors related to clustering effects of the day of the interview. Temporal aspects of conducting the fieldwork should therefore be accounted for in statistical analysis. PMID- 16040367 TI - Dust in the wind: the growing use of embalming fluid among youth in Hartford, CT. AB - This study suggests that use of embalming fluid as a mind-altering drug has been underreported. Based on a social network recruitment strategy, findings from a study in 2000 of 401 outreach worker-recruited polydrug-involved youth (ages 16 24 years) from the inner city of Hartford, CT indicate widespread (over 80% of study participants had used the drug at least once) and regular use of embalming fluid mixed with either marijuana or mint. This paper reports findings on frequency and distribution of use, experience, and consequences of use, access to the drug, and characteristics of embalming fluid users. Given the toxic substances that comprise embalming fluid, and the tendency, affirmed in the present study, of the drug to be associated with violent behavior, there is a need to recognize embalming fluid as a drug of concern among youth. PMID- 16040368 TI - Concurrent use of alcohol and cigarettes from adolescence to young adulthood: an examination of developmental trajectories and outcomes. AB - Concurrent use of alcohol and tobacco is common among adolescents, yet little is known about the developmental patterns of concurrent use, or the consequences associated with such patterns during young adulthood. Using data collected at six time points during 1985-1995 as part of an evaluation of a school-based substance abuse prevention program in California and Oregon, this study used latent growth mixture modeling to identify five distinct developmental trajectories of concurrent use of alcohol and tobacco from ages 13-23 in a cohort of 5873 individuals and compared these distinct groups with respect to demographic characteristics and young adult outcomes (at age 23 and age 29). Results suggest that while it is common during adolescence to drink but not smoke, it is very unusual to smoke and not drink. Compared to young people who smoked and drank consistently throughout their teens and early twenties, those who drank consistently but smoked only occasionally or dramatically decreased their smoking over time had lower rates of deviant behavior and predatory violence at age 23 and were less likely to have a history of arrest and substance use problems by age 29. This close examination of concurrent use of alcohol and cigarette use from ages 13-23 further accentuates the importance of curbing smoking behavior among adolescents before it becomes habitual. PMID- 16040369 TI - Physician beliefs about substance misuse and its treatment: findings from a U.S. survey of primary care practitioners. AB - Primary care physicians are in a unique position to detect patients with substance use related problems and initiate treatment at an early stage when it may be more successful. To evaluate current knowledge, attitudes, and practices of primary care physicians in the United States regarding the detection of substance use related problems, a national telephone survey of practicing primary care physicians was conducted in 1999. A total of 648 randomly selected family and general practitioners, internists, pediatricians, and OB-GYNs were interviewed regarding their perceived preparation to diagnose selected health conditions, their perceived difficulty discussing selected health conditions with patients, and their perceptions of the effectiveness of available treatments for selected health conditions. Findings suggest that primary care physicians in the United States perceive themselves as being less prepared to diagnose substance "abuse" than other chronic conditions. They additionally find it more difficult to discuss this topic with their patients, and are more skeptical about the effectiveness of available treatments. Because the social, economic, and health consequences of untreated substance misuse negatively affect the quality of life for so many patients and their families, developing new approaches to improving the attitudes and screening behaviors of primary care providers vis-a-vis substance use disorders should continue to be an important priority. PMID- 16040370 TI - Single vs. multiple drug prevention: is more always better?: a pilot study. AB - OBJECTIVE: The purpose of this study was to compare the effects of a single drug, i.e., alcohol, against a multiple drug preventive intervention. METHODS: A controlled trial was conducted with 448 8th grade students (mean age = 13 years old) from an inner-city middle school (n = 216) and a rural junior high school (n = 232) in 2000-2001. Students were randomized within school, and 3-month post intervention follow-up data were collected. RESULTS: Two risk/protective factors were found to differ significantly in favor of youth receiving the single drug alcohol intervention (p's = 0.03), while the frequency of alcohol use and two additional risk/protective factors approached significance (p's < 0.10). CONCLUSION: These findings support the potential efficacy of a brief, single drug preventive intervention over a brief, multi-drug intervention in producing short term alcohol outcomes for adolescents, and indicate differential effects of interventions for subgroups of substance using youth. PMID- 16040371 TI - Self-reported effects of methadone on cigarette smoking in methadone-maintained subjects. AB - Subjects maintained on methadone evidence a high level of interest in quitting cigarette smoking. Readiness to quit may result, at least partially, from direct pharmacological interactions between methadone and brain nicotinic acetylcholine receptors. The aim of the present study was to assess: (1) self-reported changes in smoking habits after admission to a methadone maintenance treatment (MMT) program; (2) acute effects of methadone administration on smoking behavior in MMT patients. The study was conducted between May and December 2001, in two public outpatient MMT clinics located in Warsaw, Poland. The patients (41 men, 30 women) reported smoking fewer cigarettes after admission to the program. Most subjects (67.6%) changed their favorite brand of cigarettes after admission. Mean nicotine content (mg/cigarette) significantly decreased. On the other hand, the subjects did not report any effects of methadone administration on smoking parameters. The above findings suggest that initiation of MMT is associated with positive changes in smoking behavior. However, these changes may not be related to direct pharmacological interactions between methadone and nicotine. PMID- 16040372 TI - Predictors of current depressive symptoms in a sample of drug court participants. AB - Interviews, completed between March 2000 and November 2002 with Kentucky drug court participants in Lexington and Bowling Green (N = 500), participated in a cross-sectional analysis examining the associations between self-reported, current depressive symptoms and various personal characteristics and experiences from the period before drug court involvement. Depressive symptoms were measured with the Brief Symptom Inventory (BSI), and potential correlates were derived from the Addiction Severity Index (ASI), 1992 version. BSI depression scores indicated minimal-to-moderate symptoms, with a mean individual score of 0.73, on a scale from none (0) to extreme (4) symptom strength. Numerous predictor variables were significantly associated, but multiple regression analysis identified five variables as independent correlates of depressive symptoms: 1) poorer self-rated health, 2) having ever been treated in a hospital for psychological or emotional problems, 3) being troubled by family problems in the 6 months before drug court, 4) having had conflicts with nonfamily others in the 6 months before drug court, and 5) being female. If confirmed by future, prospective research, the five variables found by the multiple regression analysis may be useful in identifying and more adequately treating substance abusers with symptoms of depression. PMID- 16040373 TI - Adolescent smoking: are infrequent and occasional smokers ready to quit? AB - We examined motivation for cessation and plans to quit among 945 high school students in 2001 at three levels of smoking frequency (regular, occasional, and infrequent). Surveys were completed at six Illinois high schools. In multivariate models, females (compared to males) and white students (compared to nonwhite students) were more likely to plan to quit, but plans to quit did not differ by smoking level. In multivariate models to predict motivation for cessation, regular smokers were significantly less motivated to quit than were occasional and infrequent smokers, and occasional smokers were less motivated than infrequent smokers; demographic variables were nonsignificant. Infrequent and occasional smokers are motivated to quit and intend to quit soon, making them an attractive target for cessation programs. PMID- 16040374 TI - Telescoping of drinking-related behaviors: gender, racial/ethnic, and age comparisons. AB - OBJECTIVE: To examine demographic differences in alcohol telescoping-the rate of self-reported movement from regular alcohol consumption to the onset of regular heavy drinking-as well as differences in ages of initiation of regular alcohol use and alcohol use to intoxication. METHOD: The present study compared the retrospective reports of 2037 clients (1252 males, 785 females) enrolled in substance user treatment facilities around the country in 1998 and 1999. RESULTS: Overall, the findings support previous research, indicating that women generally move more rapidly than men from initiation of regular alcohol use to problem use (telescoping). The findings also indicate that African-Americans generally begin regular alcohol use later than their Anglo counterparts and move more rapidly from initiation of regular alcohol use to problem use (telescoping). Examining the sample by older vs. younger age groups demonstrates that gender and racial/ethnic differences in the age of initiation of regular alcohol use and in telescoping may be limited by age group, as these patterns appear in the older but not the younger cohort. CONCLUSIONS: These findings suggest that self reported regular alcohol use, alcohol abuse, and telescoping differences vary dramatically by gender, race/ethnicity, and age cohort. Furthermore, the pattern of findings suggests that these differences are more likely the result of sociocultural than biological differences between the groups under study. PMID- 16040375 TI - Sociodemographic disparities in access to addiction treatment among a cohort of Vancouver injection drug users. AB - Vancouver's explosive HIV epidemic among injection drug users (IDUs) has received international attention due to the presence of a large needle exchange program. The role of addiction treatment has not been evaluated in this setting. We evaluated factors associated with use of addiction treatment among a prospective cohort of Vancouver IDUs. Addiction treatment was negatively associated with Aboriginal ethnicity and unstable housing, both of which have been associated with HIV infection in previous studies. These findings demonstrate low levels of addiction treatment among Vancouver IDUs and suggest that programs may need to be targeted towards specific populations with poor access. PMID- 16040376 TI - Use of dopamine agonist pergolide in outpatient treatment of cocaine dependence. AB - The dopamine agonist pergolide was evaluated in the treatment of 42 men who manifested cocaine dependence in a single-blind, 4-week-long placebo-controlled study, during 1998-1999 in Sao Paulo, Brazil. The patients were randomly assigned to two groups: the first group received pergolide (0.05-0.2 mg per day) and the second group received placebo (one to four tablets per day). Urine toxicology screens were obtained. The groups were compared in terms of depressive symptoms, "craving," use of cocaine, side effects of medications, results of urine tests, and retention in treatment. At 3 months' follow-up, the participants were reassessed. No differences were found between the two groups. PMID- 16040379 TI - BM stem cells and cardiac repair: where do we stand in 2004? AB - Adult BM stem cells are being investigated for their potential to regenerate injured tissues by a process referred to as plasticity or transdifferentiation. Although data supporting stem cell plasticity is extensive, a controversy has emerged based on findings that propose cell-cell fusion as a more appropriate interpretation for this phenomenon. A major focus of this controversy is the claim that acutely infarcted myocardium in adult hearts can be regenerated by BM stem cells. Many researchers consider the adult heart to be a post-mitotic organ, whereas others believe that a low level of cardiomyocyte renewal occurs throughout life. If renewal occurs, it may be in response to cardiac stem cell activity or to stem cells that migrate from distant tissues. Post-mortem microscopic analysis of experimentally induced myocardial infarctions in several rodent models suggests that cardiomyocyte renewal is achieved by stem cells that infiltrate the damaged tissue. For a better understanding of the possible involvement of stem cells in myocardial regeneration, it is important to develop appropriate technologies to monitor myocardial repair over time with an emphasis on large animal models. Studies on non-human primate, swine and canine models of acute myocardial infarctions would enable investigators to utilize clinical quality cell-delivery devices, track labeled donor cells after precision transplantation and utilize non-invasive imaging for functional assays over time with clinical accuracy. In addition, if stem cell plasticity is to reach the next level of acceptance, it is important to identify the environmental cues needed for stem cell trafficking and to define the genetic and cellular mechanisms that initiate transdifferentiation. Only then will it be possible to determine if, and to what extent, BM stem cells are involved in myocardial regeneration and to begin to regulate precisely tissue repair. PMID- 16040380 TI - Natural killer cell-based immunotherapeutic strategies. AB - Not until recently have Natural killer (NK) cells stepped out of the shadow of T cells to be considered for cellular therapy of malignant diseases. This evolution has been facilitated by the discovery of specific receptors on NK cells that interact with HLA molecules on target cells but also the discovery of specific activating receptors. Since NK cells represent only about 10% of the lymphocyte population in blood, separation and enrichment are important steps if NK cells are to be used clinically. This is of particular consideration in the setting of allogeneic NK cell infusion where contaminating T-cells could potentially induce graft versus host disease. This review will describe the requirements for NK cells to recognize target cells, their ex vivo expansion and potential therapeutic applications. PMID- 16040381 TI - IL-15: targeting CD8+ T cells for immunotherapy. AB - IL-15 is a pleiotropic cytokine that plays an important role in both the innate and adaptive immune system. IL-15 promotes the activation of neutrophils and macrophages, and is critical to DC function. In addition, IL-15 is essential to the development, homeostasis, function and survival of natural killer (NK) cells, NK T (NKT) cells and CD8+ T cells. Based on these properties, IL-15 has been proposed as a useful cytokine for immunotherapy. It is currently being investigated in settings of immune deficiency, for the in vitro expansion of T and NK cells, as well as an adjuvant for vaccines. In this paper, we will review the targeting of IL-15 for immunotherapy, with a particular emphasis on its effects on CD8+ T cells. PMID- 16040382 TI - Mesenchymal stem cells: progress toward promise. AB - Despite having access to embryonic stem cells, many laboratories choose to study adult stem cells, not because of philosophical reasons but because of the practical aspects and day-to-day progress necessary for developing cellular therapeutics. There is certainly the ethical desire and responsibility to provide patients with therapies where few options exist. Multipotential cells have been isolated from adult tissues in many laboratories, characterized and their multipotentiality examined. Mesenchymal stem cells (MSC) can be isolated from several tissues but easily accessible BM seems to be the most common source. These adult stem cells may not be as 'powerful' or diverse as embryonic stem cells may one day become, but at present they offer many advantages for developing cellular therapeutics: ease of isolation, expansion potential, stable phenotype, shippability, and compatibility with different delivery methods and formulations. Their potential use as cellular therapeutics has prompted the investigation of interactions of allogeneic MSC with the immune response. The great importance of cardiovascular medicine has demanded that MSC also be tested in this discipline. We believe MSC continue to provide a substantial scientific and therapeutic opportunity, and have reviewed some of the recent developments in the field. PMID- 16040383 TI - Cost-effective manufacture of an allogeneic GM-CSF-secreting breast tumor vaccine in an academic cGMP facility. AB - BACKGROUND: GM-CSF-secreting, allogeneic cell-based cancer vaccines have shown promise for the treatment of a variety of solid tumors. We have now applied this approach to breast cancer. The aim of these studies was to optimize expansion parameters, qualify the manufacturing process, and establish expected outcomes for cGMP-compliant manufacturing of two GM-CSF-secreting breast tumor cell lines. METHODS: The variables affecting the efficiency of expanding and formulating two allogeneic GM-CSF-secreting cell lines, 2T47D-V and 3SKBR3-7, were systematically evaluated. Production criteria investigated included alternative cell culture vessels (flasks vs. cell factories), centrifugation time and speed variables for large volume cell concentration, cell seeding density, the minimal concentration of FBS required for maximal cell expansion, and the dose and timing of irradiation in relation to cryopreservation. RESULTS: These studies demonstrate that, in comparison with standard 150-cm2 tissue culture flasks, Nunc 10-Stack Cell Factories are a more efficient and practical cell culture vessel for vaccine cell line manufacture. Centrifugation optimization studies using the COBE 2991 Cell Processor established that a speed of 2000 r.p.m. (450 g) for 2 min reliably concentrated the cells while maintaining acceptable viability and bioactivity. Radiation studies established that lethal irradiation prior to cryopreservation does not compromise the quality of the product, as measured by post-thaw cell viability and GM-CSF cell line-specific secretion levels. Finally, studies aimed at optimizing the production of one vaccine cell line, 3SKBR3-7, demonstrated that seeding the cells at a higher density and maintaining them in half the initial concentration of FBS maximized the yield of bioactive cells, resulting in significant cost savings. DISCUSSION: A manufacturing process that simultaneously maximizes cell yield, minimizes cell manipulation and maintains vaccine cell potency is critical for producing cell-based cancer vaccines in an academic setting. These studies define a feasible, reproducible and cost-effective methodology for production of a GM-CSF-secreting breast cancer vaccine that is cGMP compliant. PMID- 16040384 TI - Wilms' tumor 1 (WT1) gene in hematopoiesis: a surrogate marker of cell proliferation as a possible mechanism of action? AB - BACKGROUND: Wilms' tumor 1 (WT1) gene expression is seen in a significant number of cases of human neoplasia; however, the mechanism of action remains to be clarified. We hypothesized that WT1 gene is a surrogate marker of proliferation in normal hematopoietic cells and leukemias. While we and others have recognized its value as a tool for the detection of minimal residual disease (MRD), the objective of this study was to confirm our hypothesis regarding normal. METHODS: Samples from healthy donors (n=16) and UC blood (n=9) were cultured in Methocult for 21 days. Colonies were analyzed on days 7, 14 and 21 by RT-PCR for WT1 gene expression. Our positive controls were samples from patients with leukemia (n=91). Negative controls were from normal volunteers without stimulation (n=26). RESULTS: Results showed a statistically significant difference (P<0.0001) between cultured groups, with the highest level of WT1 gene expression in the positive controls and on day 14, when cells are at their maximal proliferation. DISCUSSION: In conclusion, WT1 gene expression in the proliferating colonies was highest on day 14, although less than in leukemia samples, confirming our hypothesis that WT1 gene is a surrogate marker of proliferation, not only in leukemogenesis but also, to a lesser degree, in normal cell proliferation. PMID- 16040385 TI - B-cell expansion in the presence of the novel 293-CD40L-sCD40L cell line allows the generation of large numbers of efficient xenoantigen-free APC. AB - BACKGROUND: CD40-activated B lymphocytes have been used successfully as potent APC for the induction of T-cell responses. However, the 3T3-CD40L cell line, regularly used for engagement of CD40 on the B-cell surface, is a potential source of xenoantigens. This may affect the specificity of T cells stimulated with CD40-activated B cells, especially when generation of T-cell lines specific for endogenously processed Ag is desired. METHODS: To develop a system that allows efficient expansion of B cells in the absence of sources of xenoantigens, we created a human 293-CD40L-sCD40L cell line that produces soluble CD40L and expresses CD40L on the cell surface. B cells from patients with hematologic malignancies were expanded on the 293-CD40L-sCD40L cells and used for stimulation of either naive or in vivo primed donor T cells in three HLA-identical patient donor combinations. RESULTS: The 293-CD40L-sCD40L cell line was able to stimulate B-cell growth with an efficiency superior to that of the commonly used 3T3-CD40L cell line. In all cases T-cell lines and, subsequently, T-cell clones were generated that showed reactivity against patient and not donor B cells, suggesting their specificity for minor histocompatibility antigens (mHAg). DISCUSSION: B cells activated with GMP grade 293-CD40L-sCD40L can be used in a variety of applications. In particular, they may be suitable for ex vivo stimulation of T cells prior to donor lymphocyte infusion (DLI), which may enhance its graft versus leukemia (GvL) effect. PMID- 16040386 TI - Implantation of genetically manipulated BM cells for cardiac repair. PMID- 16040388 TI - Selective allodepletion. PMID- 16040389 TI - Improving immune reconstitution while preventing GvHD in allogeneic stem cell transplantation. AB - Allogeneic hematopoietic stem cell transplantation (HSCT) is the treatment of choice for many hematologic malignancies and inherited disorders of the hematopoietic system. Ex vivo T-cell depletion (TCD) of the graft and post transplantation immunosuppression efficiently prevents the development of GvHD in no- MHC-identical settings. However, the consequence of these non-specific strategies is a long-lasting immunodeficiency associated with increased incidence of disease relapse, graft rejection and reactivation of viral infections. Donor lymphocyte infusion, which is used for treating leukemic relapse after allogeneic HSCT, can result in severe GvHD. Several strategies are being optimized specifically to inactivate anti-host T cells while preserving anti-leukemic or anti-microbial immunocompetence. Based on the ex vivo or in vivo elimination of anti-host T cells, or on the modulation of their anti-host activity, these approaches, which have been explored extensively in pre-clinical studies and tested in some preliminary clinical trials, are discussed in this paper. PMID- 16040390 TI - Selective depletion strategies in allogeneic stem cell transplantation. AB - Despite improved prophylaxis and treatment, GvHD remains a major limitation to optimal allogeneic stem cell transplantation. Ex vivo selective depletion (SD) is a strategy to prevent GvHD, in which host-reactive donor lymphocytes are selectively eliminated from a PBSC allograft while useful donor immune function is preserved. The elimination of alloreactive and thereby GvHD-mediating T cells has been shown to be feasible in both pre-clinical and more recently clinical studies. However, SD techniques and the translational research needed for clinical application are still under development. Here we summarize and discuss the following aspects of the SD approach: selection of an appropriate allogeneic stimulator; the responder population; the alloresponse; methods for removal of alloreacting T cells; product testing; clinical considerations. Our review highlights the diversity of possible approaches and the need to develop different techniques for specific clinical applications. PMID- 16040391 TI - Selective elimination of alloreactivity from immunotherapeutic T cells by photodynamic cell purging and memory T-cell sorting. AB - Allogeneic stem cell transplantation (alloSCT), especially in the mismatched setting, carries a high risk of life-threatening GvHD because of activation of donor T cells by Ag present on host cells. Removal of mature donor T cells can prevent GvHD but leads to delayed immune reconstitution, and an increased incidence of opportunistic infections and disease relapse. These findings demonstrate the vital role of donor T cells in providing graft-versus-tumor (GvT) and anti-pathogen effects as well as facilitating immune reconstitution. It has been well documented that GvHD can be separated from GvT effects, making it possible potentially to eliminate GvHD while preserving the immunotherapeutic benefits of donor T cells. Over the past decade, major attempts have been made to reduce GvHD incidence without loss of GvT effect, especially in the haplo identical setting. Novel techniques to deplete host-reactive donor T cells selectively have been explored. This review focuses on the use of the photodynamic cell purging (PDP) process and of sorting memory T cells for the selective elimination of alloreactivity. Minimizing the threat of GvHD while maximizing the beneficial GvT effect would broaden the scope and effectiveness of alloSCT. PMID- 16040392 TI - Elimination of alloreactive T cells using photodynamic therapy. AB - GvHD, the most important cause of morbidity and mortality after allogeneic stem cell transplantation, depends primarily on the ability of a donor T-cell subset to react to immunogenic host Ag. Recently developed culture conditions and treatment strategies may bring us closer to the selective elimination of such alloreactive T cells, often considered the holy grail of transplantation. Among the various therapeutic modalities, photodynamic therapy (PDT) offers a biological and global approach to the eradication of unwanted allo-activated T cells by combining mitochondrial targeting, P-glycoprotein inhibition and reactive oxygen species production. Indeed, the high potency of PDT against malignant cells has been harnessed to exert selective and extensive elimination of alloreactive T-cell subsets mediating GvHD, while preserving resting T cells with the ability to reconstitute the immune system for GvL activity and prevent or suppress viruses and fungi. The present paper reviews the basis of the PDT strategy, and the methodology employed. In vitro and in vivo studies that formed the proof of principle as a basis for human studies to investigate the clinical potential of PDT in the context of GvHD will be presented together with insights into future clinical applications of this versatile treatment platform. PMID- 16040393 TI - Modulation of GvHD by suicide-gene transduced donor T lymphocytes: clinical applications in mismatched transplantation. AB - In allogeneic hematopoietic cell transplantation (allo-HCT), donor lymphocytes play a central therapeutic role in both GvL and immune reconstitution. However, the full exploitation of these therapeutic properties is limited by the occurrence of GvHD. Different strategies have been investigated to obtain all the benefits derived from donor lymphocytes while avoiding the risk of GvHD. The genetic engineering of donor lymphocytes with the herpes simplex virus-thymidine kinase (HSV-TK) suicide gene confers the ability to modulate GvHD by invivo ganciclovir-induced elimination of the transduced cells. The suicide-gene strategy has applications in both donor lymphocyte infusion (DLI) for disease relapse and in add-back infusions after T-cell depleted allo-HCT. TK cell DLI resulted in anti-tumor activity in a relevant proportion of treated patients. Haplo-identical stem cell transplantation (haplo-HCT) is a promising therapeutic option for patients with high risk hematologic malignancies lacking an HLA matched donor. However, the profound T-cell depletion required to overcome the risk of lethal GvHD has been associated with a marked delayed T-cell recovery with a prolonged risk of post-transplant viral, fungal and other opportunistic infections. TK cell add-backs efficiently promote early immune reconstitution after haplo-HCT and prevent disease relapse, providing a unique tool for the control of GvHD. The genetic manipulation of donor lymphocytes with a suicide gene is a promising strategy to increase feasibility and safety of allo-HCT. PMID- 16040394 TI - Improving the ex vivo retroviral-mediated suicide-gene transfer process in T lymphocytes to preserve immune function. AB - The retroviral-mediated transfer of a suicide gene into donor T cells has been proposed as a method to control alloreactivity after hematopoietic stem cell (HSC) transplantation. Gene-modified cells (GMC) may be infused into the patient either at the time of transplantation, together with a T-cell depleted HSC graft, or after transplantation, as a donor lymphocyte infusion. Administration of a so called pro-drug activating the "suicide" mechanism only after occurrence of GvHD should selectively destroy the alloreactive GMC in vivo, eventually leading to GvHD abrogation. Although phase I-II clinical trials provided vital proof of the principle of GvHD control by suicide-gene therapy, this approach is still suboptimal. Indeed, current gene transfer strategies rely on gamma-retroviral vectors that require extensive T-cell activation and expansion for efficient transduction. Both in vitro and in vivo studies have shown that the activation, cell expansion, transduction and selection steps lead to TCR repertoire alterations and impairment of crucial T-cell functions, such as alloreactivity and anti-EBV reactivity. Thus, improvements of the suicide-gene transfer processes are required in order to preserve T-cell function. This could be achieved by using CD3/CD28 co-stimulation and immunomagnetic selection of transduced cells. In future clinical trials, lentiviral vectors may prove to be a better alternative to gamma-retroviral-mediated gene transfer, by reducing the need for prolonged ex vivo culture. PMID- 16040395 TI - Utilizing regulatory T cells to control alloreactivity. AB - Effective resetting of the immune system cannot be achieved by non-specific immunosuppression. Instead, novel strategies aim at harnessing the body's natural tolerance mechanisms to rectify an Ag-specific response without disturbing other immune functions. Fine-tuning of the balance between Ag-specific effector and regulatory T (Tr) cells is a promising strategy that requires detailed understanding of the differentiation and expansion pathways of the relevant Tr cell subsets. Here we review recent developments regarding the control of alloreactivity by induction and expansion of Tr cells. T-cell activation in the presence of tolerogenic APC and cytokines leads to the induction of Tr cells, which can mediate tolerance through cytokine-dependent and/or contact-dependent mechanisms. Better understanding of the mechanisms of immune regulation mediated by Tr cells may enable fine-tuning of specific immune responses and pave the way for novel therapeutic approaches. PMID- 16040396 TI - Therapeutic potential of CD4+ CD25+ regulatory T cells in allogeneic transplantation. AB - The subpopulation of CD4+ CD25+ immunoregulatory T cells constitutes less than of the entire CD4+ T-cell pool in mice and 2% in humans. These cells play a crucial role in the control of autoimmune processes. More recently, in vitro and in vivo data also indicate that CD4+ CD25+ immunoregulatory T cells can regulate alloreactivity. This renders them good candidates for innovative strategies in the field of transplantation. Inducing a state of immune tolerance with immunoregulatory T cells would alleviate the need for immunosuppression, and the occurrence of late allograft failure represents a major goal of transplantation immunology. Here we discuss how these naturally occurring CD4+ CD25+ immunoregulatory T cells can be used to modulate alloreactivity in hematopoietic stem cell and solid organ transplantation. PMID- 16040397 TI - Methods for isolating circulating epithelial cells and criteria for their classification as carcinoma cells. AB - Novel assay methods developed for the isolation and characterization of circulating tumor cells (CTC) of epithelial origin offer the potential of markers for the non-invasive gathering of clinical information relevant to the diagnosis, evolution and treatment of carcinoma. Of the numerous techniques currently used to analyze CTC, slide-based assays are perhaps the most common. While traditional combined immunocytochemical/brightfield microscopy systems continue to be the most frequently employed, fluorescence-based analysis is gaining in importance. This is partly because fluorescence microscopy analysis of slide-mounted CTC can provide simultaneously cytogenetic as well as morphologic and multiple phenotypic information. In particular, fluorescence microscopy analysis of slide-mounted CTC can accurately determine genetic changes at the chromosomal level in patients with recurrent disease. More importantly, by identifying genetic aberrations in CTC, it becomes possible to choose those patients most likely to benefit from a given treatment. The potential of this technique has already been demonstrated by employing fluorescence in situ hybridization (FISH) methods to measure expression of the HER2/neu gene in tissue from patients with breast carcinoma for the specific purpose of identifying those patients most likely to respond to Trastuzumab targeted therapy. Here, we review the major methodologies used in the preparation and analysis of the slide-based assays. PMID- 16040398 TI - Pigpen is a cellular binding protein of therapeutic oligonucleotides. AB - BACKGROUND: Understanding the mechanism of oligonucleotide (ON) uptake and cellular distribution is important for rational design of ON-based therapeutic strategies. The aim of this study was to investigate the possible relationship between cellular distribution of ON and the protein pigpen. METHODS: In vitro interaction of ON with the protein pigpen was detected using mass spectrometry. Cellular distribution of pigpen and co-localization of pigpen with ON was studied by fluorescence microscopy in endothelial YPEN and microglial N9 cells. RESULTS: Pigpen had similar distribution patterns in endothelial YPEN and microglial N9 cells. Pigpen was localized to the cytoplasm of both cell types. In addition, pigpen distributed to nuclei, excluding the nucleoli, and concentrated along the nuclear membrane and plasma membrane. Intensely stained foci were only observed in the nucleus and cytoplasm of YPEN cells. Although co-localization of pigpen with phosphorothioate (PS) ON was not observed for the first hour after ON uptake, co-localization was observed 8 h later. DISCUSSION: These data suggest that pigpen binds therapeutic ON and thus might contribute to ON cellular distribution. PMID- 16040399 TI - Effect of prostaglandin E2, lipopolysaccharide, IFN-gamma and cytokines on the generation and function of fast-DC. AB - BACKGROUND: Recent reports have described a new strategy for differentiation and maturation of monocyte-derived DC within only 48 h of in vitro culture (fast-DC). We compared the ability of various maturation stimuli with the generation of Ag specific T-cell responses and generation of functional fast-DC. METHODS: CD14+ cells were treated with GM-CSF and IL-4 for 1 day to generate immature DC, and were then matured with either inflammatory cytokines or a combination of lipopolysaccharide (LPS) and INF-gamma. Mature DC were then used to study the effect of prostaglandin E2 (PGE2) on the stimulatory function of fast-DC. RESULTS: fast-DC were CD14- and expressed mature DC surface markers, and maintained this phenotype after withdrawing the cytokine from culture. Treatment of fast-DC with a combination of LPS and INF-gamma promoted the maturation of highly uniform fast-DC. The T-cell proliferative response to DC was enhanced by inclusion of PGE2 in the MCM-mimic (TNF-a, IL-1 a, IL-6, PGE2) cocktail. DISCUSSION: fast-DC are very effective; they not only reduce the labor, cost and time required for in vitro DC development, but may also represent a model more closely resembling DC differentiation from monocytes in vivo. PMID- 16040401 TI - Peroxisome proliferator-activated receptors and their relevance to dermatology. AB - Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear hormone receptor superfamily and are expressed in a variety of tissues including skin and cells of the immune system. They act as ligand-dependent transcription factors which heterodimerize with retinoid X receptors to allow binding to and activation of PPAR responsive genes. Through this mechanism, PPAR ligands can control a wide range of physiological processes. Based on their effects in vitro and in vivo PPAR agonists and antagonists have the potential to become important therapeutic agents for the treatment of various skin diseases. PPARs can also be activated directly by phosphorylation to have ligand-independent effects. This review will discuss the physiology of PPARs relating this to skin pathology and their role as a target for novel therapies. PMID- 16040402 TI - The in vitro activity of pentane-1,5-diol against aerobic bacteria. A new antimicrobial agent for topical usage? AB - Multi-resistance to antibiotic therapy and to biocides is becoming increasingly common, which has led to mounting concern worldwide regarding the future use of traditional antimicrobials. Diols or glycols also have antimicrobial effects. Pentane-1,5-diol has low oral toxicity, is essentially non-irritating to the skin and has high antimicrobial activities against bacteria, fungi and viruses. The effect of pentane-1,5-diol against both sensitive and multi-resistant Gram positive and Gram-negative bacteria was tested in vitro against 85 bacterial strains showing minimal inhibitory concentrations in the range of 2.5 to 15.0% (vol/vol) against both antibiotic-susceptible and multi-resistant aerobic bacteria. The exact mechanism of action is unknown but probably pentane-1,5-diol sucks water out of the bacterial cells which then collapse, a mechanism to which it is probably very difficult to develop resistance. The high activity against multi-resistant bacteria makes pentane-1,5-diol an interesting new compound for topical antimicrobial therapy in humans. PMID- 16040403 TI - Fruit acids do not enhance sodium lauryl sulphate-induced cumulative irritant contact dermatitis in vivo. AB - Combined exposure to different irritants in the workplace may lead to irritant contact dermatitis, which is the main type of occupational dermatitis among bakers and confectioners. Following previous work on "tandem irritation", a panel of healthy volunteers was exposed twice daily for 4 days to the organic fruit acids: citric, malic, and lactic acid, either alone or in tandem application with 0.5% sodium lauryl sulphate (SLS) in a repetitive irritation test. Irritant cutaneous reactions were quantified by visual scoring and non-invasive measurement of transepidermal water loss and skin colour reflectance. Twice daily application of either citric or malic acid alone did not induce a significant irritant reaction. Combined exposure to one of the fruit acids and SLS caused marked barrier disturbance, but the latter irritant effect was smaller than that obtained by combined exposure to SLS and water. Thus, combined exposure to the above-mentioned fruit acids and SLS did not enhance cumulative skin irritation. PMID- 16040404 TI - Density of Demodex folliculorum in perioral dermatitis. AB - The role of Demodex folliculorum in perioral dermatitis is not satisfactory explained. Our purpose was to assess the density of D. folliculorum in perioral dermatitis and evaluate the relationship of the mite count to previous therapy with topical steroids. A standardized skin surface biopsy of the chin was performed in 82 female patients with perioral dermatitis and in 70 control female subjects. Patients who received previous topical steroid therapy had a significantly higher mite density than the patients who had received no topical steroids (p<0.001). In the latter group of patients, the mite density did not differ significantly from that of the control group (p=0.629). Mite density increased significantly with the length of treatment with topical steroids (p<0.001). Our results suggest that increased density of D. folliculorum in perioral dermatitis is a secondary phenomenon, associated with topical steroid therapy. PMID- 16040405 TI - Studies of transforming growth factors beta 1-3 and their receptors I and II in fibroblast of keloids and hypertrophic scars. AB - Keloids are benign skin tumours occurring during wound healing in genetically predisposed patients. They are characterized by an abnormal deposition of extracellular matrix components, particularly collagen. There is uncertain evidence that transforming growth factor-beta (TGFss) is involved in keloid formation. Therefore we investigated the expression of TGFss1, 2 and 3 and their receptors in keloids, hypertrophic scars and normal skin. Dermal fibroblasts were obtained from punch biopsies of patients with keloids and hypertrophic scars and from normal skin of healthy individuals. Total RNA was isolated and the expression of TGFss1, 2 and 3 and of TGFss receptors I and II (TGFssRI and II) was analysed by real-time PCR using the Lightcycler technique. Our data demonstrate significantly lower TGFss2 mRNA expression in hypertrophic scar fibroblasts as compared with fibroblasts derived from keloids and normal skin (p<0.05). In contrast, TGFss3 mRNA expression was significantly lower in keloid fibroblasts in comparison with fibroblasts derived from hypertrophic scar and normal skin (p<0.01). TGFssRI mRNA expression was significantly decreased in hypertrophic scar fibroblasts (p<0.01) and TGFssRII mRNA expression was decreased in keloids compared with hypertrophic scar fibroblasts (p<0.001). The ratio of TGFssRI/TGFssRII expression was increased in keloids compared with hypertrophic scar and normal skin fibroblasts. As recently supposed, an increased TGFssRI/TGFssRII ratio could promote fibrosis. Therefore our data support a possible role of TGFssRI and TGFssRII in combination with a certain TGFss expression pattern as fibrosis-inducing factors in keloids. PMID- 16040406 TI - Gene mutation analysis in five cases of dermatofibrosarcoma protuberans using formalin-fixed, paraffin-embedded tissues. AB - Fusion of the collagen type I a 1 (COL1A1) gene with the platelet-derived growth factor B-chain (PDGFB) gene has been pointed out in dermatofibrosarcoma protuberans. Various exons of the COL1A1 gene have been shown to be involved in the fusion with exon 2 of the PDGFB gene. We studied the breakpoints of the COL1A1 gene using formalin-fixed, paraffin-embedded tumour specimens from five patients with dermatofibrosarcoma protuberans (three reconfirmations and two new cases). Reverse transcriptase-PCR was performed using paraffin-embedded tissues. Nucleotide sequence analysis was carried out using the PCR products to identify the breakpoints. The COL1A1-PDGFB fusion transcripts were detected from the tumour specimens. Sequence analysis revealed that the ends of exons 18, 29, 38, 42 and 44 in the COL1A1 gene were fused with the start of exon 2 in the PDGFB. This study identified a novel COL1A1 breakpoint, namely, exon 44 of the COL1A1 gene. Detection of the aberrant fusion transcript using formalin-fixed, paraffin embedded tumour specimens is useful as a diagnostic aid for dermatofibrosarcoma protuberans in cases where fresh or frozen samples of tumour tissue are not available. PMID- 16040407 TI - Women with palmoplantar pustulosis have disturbed calcium homeostasis and a high prevalence of diabetes mellitus and psychiatric disorders: a case-control study. AB - Palmoplantar pustulosis is characterized by pustule formation in the acrosyringium. Nearly 50% of palmoplantar pustulosis sera produce immunofluorescence of the palmar papillary endothelium from healthy subjects, but also of the endothelium of normal parathyroid gland. With a case-control design the levels of calcium and parathyroid hormone in serum were measured in 60 women with palmoplantar pustulosis and 154 randomly selected population-based control women. One-third of the controls had been smokers, whereas 95% of the cases were or had been smokers. Mean age-adjusted serum calcium was increased in the patients compared with the controls (2.43 vs 2.36 mmol/l; p<0.0001), whereas the parathyroid hormone concentration was suppressed (23.2 vs 31.1 ng/l; p<0.0001). The plasma levels of parathyroid hormone-related protein were normal in patients but there was a strong expression of this protein in the acrosyringium both in palmoplantar pustulosis and control skin. As even a marginal elevation of serum calcium is associated with an increased risk for diabetes, cardiovascular disease and psychiatric disease, we analysed the risk for these disorders in palmoplantar pustulosis patients compared with that in the control group. Both diabetes mellitus and psychiatric disorders were associated with palmoplantar pustulosis with an odds ratio of 8.7 (95% CI 3.3-22.8) and 5.6 (95% CI 2.2-14.4), respectively. Palmoplantar pustulosis is a complex disease with an increased risk for several non-dermatological disorders. The role of the mildly increased serum calcium for the high risk for diabetes and depression deserves to be studied. PMID- 16040408 TI - Paraneoplastic pemphigus associated with non-Hodgkin B-cell lymphoma and good response to prednisone. AB - Paraneoplastic pemphigus is a life-threatening autoimmune bullous disease associated with neoplasia, generally of lymphoid origin. Immunosuppressive therapy is often disappointing and there are only a few reports of patients surviving more than 2 years. These cases were generally associated with benign neoplasms. We report here the case of a patient with paraneoplastic pemphigus associated with non-Hodgkin B-cell lymphoma who had a surprisingly good response to systemic corticosteroids and remains free of lesions more than 3 years later despite progression of her neoplasm. PMID- 16040409 TI - Comparison of tazarotene 0.1% gel plus petrolatum once daily versus calcipotriol 0.005% ointment twice daily in the treatment of plaque psoriasis. AB - Tazarotene and calcipotriol are both effective in the treatment of psoriasis. An investigator-blind, bilateral comparison of 44 lesion pairs in 19 patients was conducted to evaluate the efficacy, side effects and duration of therapeutic effects of once-daily tazarotene 0.1% gel plus petrolatum with twice-daily calcipotriol 0.005% ointment in plaque psoriasis. It consisted of a 12-week treatment phase, followed by a 4-week post-treatment observation phase. At the end of the treatment phase, tazarotene-petrolatum was as effective as calcipotriol in both objective and subjective overall efficacy assessment. Calcipotriol had a significantly greater effect in reducing erythema than tazarotene-petrolatum at weeks 2-8. At week 16, tazarotene-petrolatum demonstrated a significantly better maintenance effect in all parameters. Local irritation was noted only in tazarotene-petrolatum-treated lesions. Once-daily tazarotene 0.1% gel plus petrolatum was as effective as twice-daily calcipotriol 0.005% ointment in the treatment of plaque psoriasis, but had a better maintenance effect after the cessation of therapy. PMID- 16040410 TI - Thymus is enlarged in children with current atopic dermatitis. A cross-sectional study. AB - Atopic dermatitis is a common skin disorder of unknown aetiology with peak incidence in early childhood. The disease is associated with peripheral T-cell accumulation in the skin. The thymus is a key organ of the cellular immune response early in life. We hypothesized that atopic dermatitis is associated with an unbalanced establishment of the peripheral T-lymphocyte system. This cross sectional study was performed to compare thymus sizes in patients with atopic dermatitis and healthy controls. Thirty-seven children with current atopic dermatitis were enrolled and compared with 29 healthy controls. An interview and medical examination were performed by one doctor, an ultrasound scan was performed within 3 days of the examination, and the thymus index, a marker of thymus size, was measured. The thymus index was on average 32% higher (95% CI 3% 67%) in children with active atopic dermatitis compared with healthy controls. It declined with age in both children with atopic dermatitis and healthy controls, but the reduction in size was only significant for healthy controls. We demonstrate increased size of thymus among children with active atopic dermatitis compared with healthy controls. The larger size of thymus is compatible with increased thymic activity and emission of T lymphocytes. PMID- 16040411 TI - Stable incidence of atopic dermatitis among children in Denmark during the 1990s. AB - An increase in the prevalence of atopic dermatitis (AD) has been reported since the 1960s. The increase could be due to many factors including a genuine increase of incidence or duration of AD. We decided to study if the increasing trend persisted during the 1990s by comparing the cumulative incidence of AD in 1993 and 1998. Further, we studied the severity and management of AD among children. Two samples of children born in Denmark were drawn from the Danish Medical Birth Register. In the 1993 and 1998 studies a mailed questionnaire with identical questions concerning AD was sent out. In the 1998 follow-up study the questionnaire included a severity score and questions concerning management of AD. In the 1993 study the cumulative incidence of AD at age 7 was 18.9% and in 1998 it was 19.6%. There was no difference in the age-adjusted AD incidence in the 5-year observation period. In the 1998 study, 81% had mild to moderate AD, 90% had been seen by a doctor at least once, 36% had mainly been treated by a dermatologist, and 2% had been hospitalized. It should be kept in mind that we base most of our common knowledge of the disease on AD patients selected for management in dermatology clinics and departments. PMID- 16040412 TI - Acceptance and outcome of herpes simplex virus type 2 antibody testing in patients attending an STD clinic--recognized and unrecognized infections. AB - The majority of herpes simplex virus type 2 (HSV-2) genital infections are asymptomatic. We wanted to evaluate the acceptance of HSV-2 antibody testing among people attending an STD clinic and to estimate, after counselling, the percentage of recognized and unrecognized HSV-2 infections. First visitors to an STD clinic were invited to participate by answering a questionnaire and taking a blood test for HSV-2 antibodies. HSV-2 seropositive individuals, who were unaware of having genital herpes, were offered an HSV-2 counselling visit and follow-up. Of 1769 patients offered testing, 57% accepted. Of 152 (15%) HSV-2 seropositive individuals, 41% had a self-reported history of genital herpes, approximately 30% had genital symptoms and 30% had no genital symptoms. The percentage of patients reporting genital symptoms was much higher in HSV-2 seropositives (45%) without a history of genital herpes than in an HSV-2 seronegative group (28%). HSV-2 antibody testing should be performed generously in all cases of uncharacteristic genital symptoms. PMID- 16040413 TI - Anti-citrulline antibodies in psoriatic patients with and without arthritis. PMID- 16040414 TI - Low-dose dapsone in chronic idiopathic urticaria: preliminary results of an open study. PMID- 16040415 TI - Onchocerciasis diagnosed in Italy. PMID- 16040416 TI - Chromoblastomycosis: an unusual diagnosis in Europe. PMID- 16040417 TI - Onychomycosis due to Aspergillus tamarii in a 3-year-old boy. PMID- 16040418 TI - Neutrophilic lobular panniculitis with non-rheumatoid arthritis. PMID- 16040419 TI - Allergic contact dermatitis due to an insect repellent: double sensitization to picaridin and methyl glucose dioleate. PMID- 16040420 TI - Dermatophytes in a swimming pool facility: difference in dermatophyte load in men's and women's dressing rooms. PMID- 16040421 TI - Reactive angioendotheliomatosis associated with myelodysplastic syndrome. PMID- 16040422 TI - Multiple eruptive dermatofibromas occurring in a patient with myelodysplastic syndrome. PMID- 16040423 TI - Schnitzler's syndrome without monoclonal gammopathy. PMID- 16040424 TI - Late-onset familial onychodystrophy heralding incontinentia pigmenti. PMID- 16040425 TI - Fibrous hamartoma of infancy: a case report. PMID- 16040426 TI - Severe and unresponsive HIV-associated alopecia areata successfully treated with thalidomide. PMID- 16040427 TI - Zosteriform morphea: a new pattern. PMID- 16040428 TI - Anaphylactic reaction caused by the UVA absorber disodium phenyl dibenzimidazole tetrasulfonate. PMID- 16040429 TI - Symmetrical cutaneous leishmaniasis. PMID- 16040430 TI - Incidental finding: a penile cutaneous horn. PMID- 16040434 TI - Quality of life in mothers of speech impaired children. AB - Today, health-related quality of life (HRQOL) is an important outcome parameter in medicine. As a mother's well-being is important for the development of her child it is assumed that her HRQOL influences her child. In children with developmental disorders mothers may experience a reduction of their HRQOL. This study focused on HRQOL in mothers of speech impaired children. In 91 mothers the SF-36 questionnaire revealed lower scores of all subscales except for 'bodily pain' and 'mental health'. Although the data allow for no conclusion on whether the speech impairment is the reason or the consequence of a reduced HRQOL, the results indicate the necessity to take into account the mothers' well-being when dealing with speech impaired children. PMID- 16040435 TI - Voice-related quality of life in organic and functional voice disorders. AB - The voice-related quality of life (V-RQOL) questionnaire was proposed as a disease specific measure of patients' subjective burden elicited by a voice disorder. The purpose of this study was to investigate the influence of gender and aetiology (benign organic versus functional) on V-RQOL results. Fifty-two German-speaking patients completed the questionnaire without prior information about their individual diagnosis. Results allow for the conclusion that women score lower indicating a poorer voice status. Differences between patients diagnosed with an organic or a functional voice disorder revealed no universal difference. This has to be investigated in future, especially against the degree of the voice disorder, the individual personality, health belief and social background. PMID- 16040436 TI - Voice symptoms of call-centre customer service advisers experienced during a work day and effects of a short vocal training course. AB - Occupational voice users often suffer from voice symptoms to varying extents. The first goal of this study was to find out how telephone customer service advisers experience voice symptoms at different moments of the working day. The second goal was to investigate the effects of a short vocal training course arranged for telephone workers. The results indicate that although the subjects did not suffer from severe voice problems, the short vocal training course significantly reduced some of the vocal symptoms they had experienced. The results suggest that systematic consultation and training for occupational voice users in the field of occupational voice care would be advantageous. PMID- 16040437 TI - Comprehension of before and after in frontotemporal degeneration. AB - Frontotemporal lobar degeneration (FTLD) is a dementia syndrome with behavioural problems and deterioration of language functions. The other cognitive domains are relatively preserved for at least the first two years of the disease. We studied the ability of patients with FTLD to understand before and after constructions expressed in Finnish. Ten patients and eight controls were presented eight sentences with different relations and asked questions to test their comprehension. The results showed that the patients generally found after constructions more difficult. The difficulty was not, however, related to normal ageing, as was shown by the performance of the control group. PMID- 16040438 TI - Artificially lengthened and constricted vocal tract in vocal training methods. AB - It is common practice in vocal training to make use of vocal exercise techniques that involve partial occlusion of the vocal tract. Various techniques are used; some of them form an occlusion within the front part of the oral cavity or at the lips. Another vocal exercise technique involves lengthening the vocal tract; for example, the method of phonation into small tubes. This essay presents some studies made on the effects of various vocal training methods that involve an artificially lengthened and constricted vocal tract. The influence of sufficient acoustic impedance on vocal fold vibration and economical voice production is presented. PMID- 16040439 TI - Communication aids for people with aphasia. PMID- 16040440 TI - Evaluation of muscular activity, local muscular fatigue, and muscular rest patterns among dentists. AB - In a previous study, dentists reported very high scores for perceived physical workload, but only low to moderate scores for the musculoskeletal system. This is difficult to explain when other occupational groups in the dental services are compared, and is the main reason why the present study was performed. To measure muscular activity, a surface electromyography (sEMG) study was done, and included the subjects who reported neck and shoulder complaints in the previous study. A portable sEMG system (MyoGuard) was used to collect a myoelectric signal on-line and analysis of the myoelectric signal in a computer. sEMG was recorded from both trapezius muscles for approximately 4 h during an ordinary working day. Twenty seven dentists participated in the study. The results show accumulated rest% fairly close to that of female cashiers and supermarket employees and increased average rectified value percent (ARV%) during work that could contribute to the very high workload perceived by dentists. PMID- 16040441 TI - Evaluation of some properties of two fiber-reinforced composite materials. AB - OBJECTIVE: Water sorption, flexural properties, bonding properties, and elemental composition of photopolymerizable resin-impregnated fiber-reinforced composite (FRC) materials (everStick C&B and BR-100) (FPD) were evaluated in this study. MATERIAL AND METHODS: Bar-shaped specimens (2 x 2 x 25 mm) were prepared for water sorption and flexural strength testing. The specimens (n = 6) were polymerized either with a hand light-curing unit for 40 s or, additionally, in a light-curing oven for 20 min and stored in water for 30 days. Water sorption was measured during this time, followed by measurements of flexural strength and modulus. A shear bond strength test was performed to determine the bonding characteristics of polymerized FRC to composite resin luting cement (Panavia-F), (n = 15). The cement was bonded to the FRC substrate and the specimens were thermocycled 5000 times (5-55 degrees C) in water. SEM/EDS were analyzed to evaluate the elemental composition of the glass fibers and the fiber distribution in cross section. RESULTS: ANOVA showed significant differences in water sorption according to brand (p < 0.05). Water sorption of everStick C&B was 1.86 wt% (hand unit polymerized) and 1.94 wt% (oven polymerized), whereas BR-100 was 1.07 wt% and 1.17 wt%, respectively. The flexural strength of everStick C&B after 30 days' water storage was 559 MPa (hand-unit polymerized) and 796 MPa (oven-polymerized); for BR-100, the values were 547 MPa and 689 MPa, respectively. Mean shear bond strength of composite resin cement to the FRC varied between 20.1 and 23.7 MPa, showing no statistical difference between the materials. SEM/EDS analysis revealed that fibers of both FRC materials consist of the same oxides (SiO2, CaO, and Al2O3) in ratios. The distribution of fibers in the cross section of specimens was more evenly distributed in everStick C&B than in BR-100. CONCLUSION: The results of this study suggest that there are some differences in the tested properties of the FRC materials. PMID- 16040442 TI - Interface evaluation after manual and ultrasonic insertion of standardized class I inlays using composite resin materials of different viscosity. AB - OBJECTIVE: The aim of the present study was to investigate the effect of manual and ultrasonic insertion of standardized class I inlays (Cerana) using three composite resin materials of different viscosity (Tetric Flow, Tetric, and Tetric Ceram) on time to seat inlays, film thickness, and filler distribution within the materials. METHODS: In a preliminary test, mean loads for manual and ultrasonic insertion were measured using the high viscosity composite resin material (Tetric Ceram). These loads were then applied with all composite resin materials to evaluate the times required to seat the inlays. In addition, film thickness was assessed using scanning electron microscopy, and filler distribution (wt% silicon, barium, ytterbium) was monitored using energy-dispersive spectroscopy. RESULTS: Ultrasonic insertion significantly reduced mean load applied to seat inlays (6.4+/-1.4 N; mean+/-SD) as compared to manual insertion (18.9+/-3.1 N; p < 0.001). Using an ultrasonic device, times for insertion values were significantly lower in the high and medium viscosity composite resin material groups compared to manual insertion (p < 0.05). The widest film thickness was recorded for the high viscosity composite resin material in combination with manual insertion (p < 0.05). However, when ultrasound was applied, there was no difference in film thickness between the three materials at any levels. Furthermore, the analysis of filler distribution revealed no significant differences between groups. CONCLUSION: Highly filled viscous composite resin materials may be used in combination with the ultrasonic insertion technique without untoward effects on film thickness or filler distribution. PMID- 16040443 TI - Perceived orofacial pain and its associations with reported bruxism and insomnia symptoms in media personnel with or without irregular shift work. AB - A standardized questionnaire was mailed to all employees of the Finnish Broadcasting Company with irregular shift work (n = 750) and to an equal number of randomly selected controls in the same company with regular 8-hour daytime work. The aims were to investigate the prevalence and severity of perceived orofacial pain (Research Diagnostic Criteria for Temporomandibular Disorders Axis II) and to analyze whether current orofacial pain was associated with reported bruxism and insomnia symptoms (Diagnostic and Statistical Manual of Mental Disorders-IV and the International Classification of Sleep Disorders Revised). The response rate in the irregular shift-work group was 82.3% (56.6% men) and in the regular daytime-work group 34.3% (46.7% men). Current orofacial pain was found overall in 19.6%, of which 88.3% had experienced the pain over 6 months. All claimed that their pain fluctuated. No subjects with chronic orofacial pain reported disabling pain, and grades III and IV were not found. Insomnia symptoms and frequent bruxism were significantly more prevalent in chronic pain grade II than in lower grades. According to logistic regression, current orofacial pain was significantly positively associated with frequent bruxism (p < 0.001), female gender (p < 0.001), and disrupted sleep (p < 0.01), and significantly negatively associated with age over 45 years (p < 0.01). Our results revealed a clear-cut association between perceived orofacial pain and reported bruxism. The association held with both chronic orofacial pain intensity and current pain. Based on the multivariate analyses, it can be concluded that disrupted sleep and bruxism may be concomitantly involved in the development of orofacial pain. PMID- 16040444 TI - Clinical comparison between two different splint designs for temporomandibular disorder therapy. AB - OBJECTIVE: To compare splint therapy in temporomandibular disorder (TMD) patients using two splint designs. MATERIAL AND METHODS: In a double-blind randomized parallel trial, 40 consenting patients were selected from the dental faculty pool of TMD patients. Two splint designs were produced: an ordinary stabilization (Michigan type) and a NTI (Nociceptiv trigeminal inhibition). The differences in splint design were not described to the patients. All patients were treated by one operator. A separate, blinded, examiner assessed joint and muscle tenderness by palpation and jaw opening prior to splint therapy, and after 2 and 6 weeks' and 3 months' splint use during night-time. The patients reported headache and TMD-related pain on a visual analog scale before and after splint use, and were asked to describe the comfort of the splint and invited to comment. RESULTS: Thirty-eight patients with mainly myogenic problems were observed over 3 months. A reduction of muscle tenderness upon palpation and self-reported TMD-related pain and headache and an improved jaw opening was seen in both splint groups (p < 0.05; paired t-test and Wilcoxon signed-ranks tests). There were no changes for TM joint tenderness upon palpation. No differences were noted between the two splint designs after 3 months for the chosen criteria of treatment efficacy (p > 0.05; Mann-Whitney U-test). CONCLUSION: No differences in treatment efficacy were noted between the Michigan and the NTI splint types when compared over 3 months. PMID- 16040445 TI - Marginal and internal adaptation of composite restorations using a resin liner on deproteinized substrate. AB - OBJECTIVE: The purpose of this study was to evaluate the use of resin liner on deproteinized substrate on marginal and internal adaptation of composite restorations. MATERIAL AND METHODS: Twenty-four recently extracted 3rd molars were selected. The crowns were sectioned in a mesio-distal direction, and the specimens were ground to expose a flat enamel area of at least 6 mm in diameter. Cavities were prepared on the central area of flattened surfaces. The specimens were randomly assigned into four groups (n = 12): SB - 1. Acid etch, 2. Single bond application (SB); NaOCl/SB - 1. acid etch, 2. 10% sodium hypochlorite solution application for 60 s (NaOCl), 3. SB; SB/PL - 1. acid etch, 2. SB, 3. protect liner F (PL); NaOCl/SB/PL - 1. acid etch, 2. NaOCl, 3. SB, 4. PL. All cavities were restored with Filtek Z250 and polished. The outer and internal margins were stained using Caries Detector (Kuraray), observed under the stereomicroscope, and transferred to a computer measurement program. The length of the gap was expressed as a percentage of the total length of the margins observed. Data (internal adaptation) were submitted to ANOVA and Tukey's test. RESULTS: There were no gaps at enamel outer margins in any specimen of any group. There was no statistically significant difference in internal adaptation between SB (18%) and NaOCl/SB (18%). NaOCl/SB/PL showed the worst internal adaptation (21%), with the higher percentage of internal gaps, statistically differing from SB/PL, which showed the best internal adaptation (12%). CONCLUSIONS: Resin liner can effectively enhance internal adaptation on demineralized dentin. However, on demineralized and deproteinized dentin it affects the internal adaptation of composite restorations negatively. PMID- 16040446 TI - Effect of xylitol-containing chewing gums on interdental plaque-pH in habitual xylitol consumers. AB - OBJECTIVE: The aim was to investigate the effect of high and low amounts of xylitol on the interdental plaque-pH, directly and after sucrose challenge, in schoolchildren with habitual consumption. MATERIAL AND METHODS: The study group consisted of 11 healthy children (10-15 years) with low caries risk and the experiment had a single-blind crossover (Latin square) design. After a 2-week run in period with a daily 4.0 g xylitol intake, the children were subjected to single-dose exposures of chewing gums with (i) paraffin (CTR; no xylitol), (ii) low-dose xylitol (LX; 2.0 g xylitol), and (iii) high-dose xylitol (HX; 6.0 g xylitol) in a randomized order separated by a washout period of 1 week. Samples of chewing-stimulated whole saliva were collected prior to and after the experimental period for determination of bacterial counts. The outcome measures were in situ plaque-pH (micro-touch method) and area under the pH curve (AUC). RESULTS: The AUC was significantly greater (p < 0.05) in the HX group compared to the LX and control groups during the first 5 min after chewing. After a 10% sucrose rinse, the interdental plaque-pH dropped in all groups but the HX regimen displayed significantly less reduction 0-5 min after chewing (p < 0.05). No significant alterations of the total viable counts or mutans streptococci levels in saliva were disclosed during the 4-week experimental period. CONCLUSIONS: The present results suggested that a high single dose of xylitol had a short and limited beneficial effect on interdental plaque-pH in habitual xylitol consumers, while a low single dose, resembling normal chewing gum use, did not differ from the control. PMID- 16040447 TI - Dental fear of Finnish children in the light of different measures of dental fear. AB - OBJECTIVES: The aims were: (1) to examine the construct and reliability of the modified Dental Subscale of the Children's Fear Survey Schedule (CFSS-DS) among children of different ages and (2) to compare the correlations between fear measured with the modified CFSS-DS, the "peak value for dental fear" (PV), and "fear of dental treatment in general" (GF). METHODS: The study sample consisted of 302, 299, 314, and 297 children aged 6, 9, 12, and 15 years, respectively. Each child received a questionnaire to be filled out at home. An explorative factor analysis with varimax rotation was performed for eight items taken from the CFSS-DS and questions on fear of pain and suction used in the mouth. For further age-specific analyses, mean values were calculated for the sum of items that loaded >0.5 on each factor. The correlations between these values and PV and GF were studied. RESULTS: The questionnaire was reliable. Two factors were revealed for each age: "treatment of dental decay" (TDD), which included fears related to invasive treatment, and "attending the dentist" (AD), which included fears related to dental visits in general. TDD explained over 50% of the variance, except among 9-year-olds. TDD mean values were higher among older children than among younger ones and correlated more strongly with PV than with GF. AD mean values were higher among younger children than among older ones and correlated more strongly with GF than with PV. CONCLUSION: The factor structures were fairly similar but the correlations between fear measures differed among children of different ages. PMID- 16040448 TI - Dental caries and associated factors in Mexican schoolchildren aged 6-13 years. AB - The objectives of the present study were to establish dental caries prevalence (percentage with caries) and experience in the primary and permanent dentition (dmft and DMFT) of 6 to 13-year-old schoolchildren in Campeche, Mexico, and to estimate the contributing roles of the likely risk indicators. A cross-sectional study was carried out in 1,644 children aged 6-13 years. Self-administered questionnaires obtained information on social, economic, behavioral, and demographic variables. The primary dentition of 1,309 children and the permanent dentition of 1,640 children were evaluated in the oral examinations. The main outcome measures were DMFT, dmft, and SiC indices. Data were modeled using logistic regression analysis. The overall caries prevalence was 77.4%, 73.6% in the primary dentition (61.6% in 6-year-olds), and 49.4% in the permanent dentition. The dmft and DMFT indices were 2.85+/-2.73 and 1.44+/-2.05, respectively (DMFT = 3.11+/-2.62 in 12-year-olds). The SiC index was 6.05 at 12 years of age. Associated variables to dental caries in both dentitions were presence of enamel defects, presence of dental plaque, low socio-economic status, female sex, and older age. Mother's schooling was negatively associated (OR = 0.95) with caries in primary dentition. Caries experience in the primary dentition (OR = 6.02) was positively associated with caries in the permanent dentition. Dental caries status in these Mexican children was closer to the goals proposed by the WHO/FDI for 2000 than previous studies. This study has identified clinical, socio-economic, and behavioral determinants for dental caries in primary and permanent dentition on Mexican schoolchildren. PMID- 16040450 TI - Part 1: Some key developments in Swedish public health research 1940-2000. PMID- 16040451 TI - Part 2: An inventory of Swedish public health research. PMID- 16040452 TI - Part 3: International evaluation of Swedish public health research. PMID- 16040454 TI - Assessing environmental health risks or net health benefits? PMID- 16040455 TI - Inventory and evaluation of Swedish public health research. PMID- 16040456 TI - The Swedish Demand-Control-Support Questionnaire (DCSQ): factor structure, item analyses, and internal consistency in a large population. AB - AIM: This paper examined the psychometric properties of the Swedish Demand Control-Support Questionnaire (DCSQ), which is a shorter and modified version of Karasek's Job Content Questionnaire (JCQ). METHODS: The study encompassed 5,227 workers born in 1953-57 from the population-based Hordaland Health Study. DCSQ, a 17-item questionnaire, covers psychological demands, decision latitude, and social support in the workplace. The workers were manually classified according to the Standard Classification of Occupations. The main statistical methods were principal component analyses, and estimation of internal consistency and the subscales' shared variance by Cronbach's coefficient a and Pearson's correlation coefficients, respectively. RESULTS: The study gave support to the tri dimensional factor structure of DCSQ. Decision latitude tended to split into skill discretion and decision authority in skill-level homogeneous samples. The specificity of the item loadings was satisfactory except for "conflicting demands". The inter-correlation of the three main subscales was weak. The internal consistency of the subscales was generally satisfactory. CONCLUSIONS: The psychometric properties of DCSQ are satisfactory. Being shorter and easier to use than the more comprehensive JCQ, DCSQ represents an important alternative, particularly if respondent burden and data-collection costs need to be minimized. PMID- 16040457 TI - Who are the "quitters"? a cross-sectional study of circumstances associated with women giving up smoking. AB - BACKGROUND: Smoking is an important preventable risk factor for cardiovascular disease, cancer, and many other diseases. Even though tobacco consumption is declining in Sweden, it is not declining in all groups. This study explored socioeconomic and psychosocial circumstances hindering or facilitating smoking cessation in three birth cohorts of women from the general population. METHODS: Between 1991 and 1996 a comprehensive questionnaire was administered to 17,319 women, 45-73 years old, from the Malmo Diet and Cancer cohort. Smoking habits were compared in relation to socioeconomic and psychosocial circumstances in three birth cohorts. RESULTS: Of these women, 44% were never smokers, 28% were ex smokers, and 28% were smokers (regular or occasional). When compared with smokers, ex-smokers were more often married, had a higher socioeconomic position, a longer education, more smoke-free surroundings, better emotional support, higher BMI, and better self-perceived health. Ex-smokers reported less work related stress and less shift work. A history of cardiovascular disease was not associated with smoking cessation. The socioeconomic differences between current and former smokers were higher for young women as compared with older birth cohorts. CONCLUSIONS: Continuing smokers and quitters differ with regard to socioeconomic and psychosocial circumstances and factors related to working life and environmental tobacco exposure. By determining who the quitters are through continued follow-up, useful insights can be gained to develop strategies to achieve successful cessation of smoking. PMID- 16040458 TI - Self-rated health in relation to age and gender: influence on mortality risk in the Malmo Preventive Project. AB - AIMS: A study was undertaken to examine whether poor self-rated health (SRH) can independently predict all-cause mortality during 22-year follow-up in middle-aged men and women. SUBJECTS AND METHODS: Data are derived from a population-based study in Malmo, Sweden. This included baseline laboratory testing and a self administered questionnaire. The question on global SRH was answered by 15,590 men (mean age 46.4 years) and 10,089 women (49.4 years). Social background characteristics (occupation, marital status) were based on data from national censuses. Mortality was retrieved from national registers. RESULTS: At screening 4,261 (27.3%) men and 3,085 (30.6%) women reported poor SRH. Among subjects rating their SRH as low, 1,022 (24.0%) men and 228 (7.4%) women died during follow-up. Corresponding figures for subjects rating their SRH as high were 1801 (15.9%) men and 376 (5.4%) women. An analysis of survival in subjects reporting poor SRH revealed an age-adjusted hazard risk ratio (HR, 95%CI) for men HR 1.5 (1.4-1.7), and for women HR 1.4 (1.2-1.6). The corresponding HR after adjusting for possible social confounders was for men HR 1.3 (1.1-1.4), and women HR 1.1 (0.9-1.4). When additional adjustment was made for biological risk factors the association for men was still significant, HR 1.2 (1.1-1.3). CONCLUSION: Poor SRH predicts increased long-term mortality in healthy, middle-aged subjects. For men the association is independent of both social background and selected biological variables. The adjustment for biological variables can be questioned as they might represent mediating mechanisms in a possible causal chain of events. PMID- 16040459 TI - Prognosis of risk factors and trends in mortality from external causes among middle-aged men in Lithuania. AB - AIMS: The aims of this study were to investigate the risk of death and time trends from external causes, and to evaluate the significance of the effects of age, period and birth cohort on suicide mortality among middle-aged men during the period 1971-2000 in Lithuania. METHODS: Random samples of men aged 45-59 years from the Kaunas Rotterdam Intervention Study (conducted in 1972-74) and Study of Multifactorial Prevention of CHD (conducted in 1977-80) were examined (n?=?6,480). The participants of the two surveys were observed until 1 January 2001. Over this time 2,841 men had died, 230 of these from external causes. The Cox proportional hazards model was used to evaluate the risk of death from external causes. Trends in mortality from external causes and average annual changes were based on logarithmic regression analysis. For assessment of the effects of age, period, and birth cohort the Poisson regression model was applied. RESULTS: The risk of mortality from external causes among men was positively related to smoking habits and arterial hypertension and negatively related to education level and total serum cholesterol concentration but there was no association with consumption of alcohol. The risk of suicide mortality was associated with family status and occupation only. Trends in mortality from all external causes showed no significant changes during the period 1971-2000. After adjusting for age and cohort effects, the period effect was statistically significant. CONCLUSION: Prognosis of risk factors for mortality from external causes and period effect on suicide mortality rates will form important parts of future research agendas. PMID- 16040460 TI - The Copenhagen County child cohort: design of a longitudinal study of child mental health. AB - BACKGROUND: Epidemiological studies of psychopathology in the first years of life are few, and the association between mental health problems in infancy and psychiatric disturbances later in life has not been systematically investigated. The aim of the present project was to study mental health problems and possibilities of intervention from infancy and onward. METHODS: The basic study population consists of a birth cohort of 6,090 children born in the year 2000 in the County of Copenhagen, the Copenhagen County Child Cohort, CCCC 2000. At stage one CCCC 2000 was established on data from the Civil Registration System, Danish national registers, and standardized, longitudinal data from the first year of living obtained by public health nurses. At stage two a subsample was assessed at 1(1/2) years of age concerning child psychiatric illness and associated factors in a case-control study nested in the cohort, including a random sample. Participation rate at stage one was 92%. PERSPECTIVES: Ongoing studies of CCCC 2000 include studies of failure to thrive, register studies, and studies of the predictive validity of public health screening. A follow-up study concerning the prevalence of psychopathology at age 5 is planned. SUMMARY: The Copenhagen County Child Cohort CCCC 2000 is a longitudinal study of mental health from infancy investigating psychopathology in early childhood. Results from this study will add to the knowledge of risk factors and course of mental health problems in childhood and contribute to the validation of the mental health screening made by public health nurses. PMID- 16040461 TI - Awareness of risk of osteoporosis may cause uncertainty and worry in menopausal women. AB - AIMS: A study was undertaken to explore how menopausal women are affected by awareness of potential risk of osteoporosis. METHODS: A qualitative interview study, including analysis of in-depth interviews with 17 women who independently gave views on risk, out of 24 women interviewed about their menopausal symptoms. The women were selected on the basis of a survey including 1261 women chosen at random, to cover a broad spectrum of Danish women, their menopausal experiences, and contact with the healthcare system. The study was part of a larger project targeting menopause. RESULTS: Awareness of osteoporosis risk caused a feeling of uncertainty and worry in some women. Only women reacting in this way seemed to act in order to prevent future fractures. The affected women were puzzled to realize that risk-reducing medication could introduce new hazards. Most of the women had heard about osteoporosis related to menopause as culturally embedded knowledge. CONCLUSIONS: Making individual women uncertain and worried must be considered a potentially serious side effect of health promotion. The findings raise the question of whether introducing healthy people to the threat of future diseases is ethically justifiable. As hormonal treatment is no longer recommended for long-term use, it is suggested that the strong link between osteoporosis and menopause should be toned down when counselling menopausal women. PMID- 16040462 TI - Are cancer patients at higher suicide risk than the general population? AB - OBJECTIVES: The aims were to test whether suicide is more prevalent among cancer patients than among the general population, whether the association changes over time, and whether the risk depends on the severity of the cancer disease. DESIGN: A register study was carried out, where standardized suicide rates for different cancer sites were analysed by sex, time periods (1965-74, 1975-84, and 1985-94), and five-year survival rates. SETTING: The setting was the Swedish population from 1965 to 1999. SUBJECTS: The cohort studied was 1,031,919 cancer cases and 2,587,152 person-years. MAIN OUTCOME MEASURES: The main outcome measure was the standardized mortality ratio (SMR) with the world population as standard population. RESULTS: Cancer patients run a higher risk of suicide than the general population. The suicide rate ratio between male cancer patients and the general population was about 2.5 in 1965-74, which thereafter reduced to about 1.5 in 1985-94. The suicide rate among female cancer patients was in excess of 2.9 in 1965-74 and 2.3 in 1985-94. In fact, a slightly higher rate for women was observed for 1985-94 than for 1975-84. There is also a strong negative association between survival rates and suicide rates, i.e. suicide rates are higher for severe cancer sites with low survival rates. CONCLUSIONS: Our findings support the hypothesis that cancer patients run greater risks of committing suicide than the general population does. The severity of the cancer increases the suicide risk. More attention to cancer patients' psychological and care situation is called for. PMID- 16040463 TI - Four indicators of socioeconomic position: relative ranking across causes of death. AB - OBJECTIVE: A study was undertaken to examine the relative ability of occupational class, education, household income, and housing conditions to discriminate all cause and cause-specific mortality-risk in Oslo, and to see if this relative ability is consistent across the 12 most common causes of death. DESIGN AND SETTING: Census records of inhabitants in Oslo 1990 aged 45 to 64 were linked to death records 1990-98 (n?=?88,159). All inhabitants were included except those who lacked census data on the independent variables. The relative index of inequality (RII) for each indicator was calculated. MAIN RESULTS: Education, occupation, and housing conditions had similar RIIs for all-cause mortality in both sexes. Household income had low RIIs, particularly in men. For the 12 most common causes of death some heterogeneity in the relative ranking between the four indicators was observed, with causes of death known to be related to early life social circumstances (stomach cancer, cardiovascular disease, chronic obstructive pulmonary disease) being particularly strongly related to education, and causes of death which were likely to be determined by adult social circumstances (violence, sudden unexpected death) being particularly strongly related to occupation and housing conditions. CONCLUSIONS: Education, occupational class, and housing conditions all seemed to discriminate all-cause mortality to a similar degree. However, the cause-specific analysis revealed a heterogeneous pattern. PMID- 16040464 TI - The AIDS epidemic in the city of Copenhagen, Denmark: potential years of life lost and impact on life expectancy. AB - AIMS: This study seeks to describe the impact of AIDS on the city of Copenhagen by estimating potential years of life lost (PYLL) before the age of 65 years and to estimate the impact of AIDS deaths on life expectancy for males and females. METHODS: All AIDS cases reported to the national AIDS surveillance register for residents in the city of Copenhagen in the period 1983-98 were included. For comparative purposes data were obtained on six other causes of death: accidents, suicide, lung cancer, ischaemic heart disease, testicular cancer, and breast cancer. RESULTS: Overall, deaths from AIDS accounted for 8% of all PYLL in men and showed an increasing tendency from 1983 to 1991, when it became the leading cause of PYLL. AIDS had most impact in men in the age group 25-44 years and accounted for 29% of all PYLL in this group at the peak in 1993, decreasing significantly after the introduction of anti-retroviral treatments to 5% of PYLL in 1998. Other leading causes of PYLL, accidents and suicide, also showed a decreasing tendency over the years, but of a much smaller magnitude than AIDS. The impact of AIDS in women was more modest. In the entire study period suicide, accidents, and breast cancer were the leading causes of PYLL in women. It was shown that AIDS deaths at the top of the epidemic in 1991-95 were responsible for a loss of 0.76 years in life expectancy for men and 0.08 years for women. CONCLUSIONS: AIDS has had a considerable impact on potential years of life lost. A significant decline in AIDS deaths has been seen since 1995 with an effect on life expectancy for men in the city of Copenhagen. PMID- 16040465 TI - Consultations for injuries by frequent attenders are found to be medically appropriate from general practitioners' perspective. AB - AIMS: A study was carried out to determine whether frequent attenders' (FAs') consultations for injuries are medically appropriate or not. METHODS: A retrospective review was undertaken of medical records in a primary healthcare centre. All injury consultations by frequent attenders and controls during a period of one year were evaluated by two general practitioners (GPs). Outcome variables were: number of injuries, chronic diseases, type of injuries, and their treatments. The GPs made a comprehensive picture of each case and evaluated whether it was medically appropriate to consult a doctor or not. RESULTS: Injuries were 7.2 times more common among frequent attenders compared with the controls. Frequent attenders had significantly more chronic diseases. Mean number of injury consultations was the same for frequent attenders with or without chronic disease. There was no difference concerning medically appropriate consultations between frequent attenders and controls. CONCLUSIONS: Consultations for injuries with frequent attenders were considered medically appropriate. Chronic diseases did not explain the higher injury incidence among frequent attenders. These findings are interesting and contradict the opinion that increased healthcare consumption by FAs is a waste of resources. Our findings may suggest that FAs are more vulnerable individuals. PMID- 16040466 TI - Is HIV control in sub-Saharan Africa losing focus?: the need for simplified, prioritized prevention strategies. PMID- 16040467 TI - The People's Health Movement: what is it and should we care? PMID- 16040468 TI - Heterotopic ossification: a review. AB - Heterotopic ossification is defined as the presence of lamellar bone at locations where bone normally does not exist. The condition must be distinguished from metastatic calcifications, which mainly occur in hypercalcaemia, and dystrophic calcifications in tumours. It is a frequent complication following central nervous system disorders (brain injuries, tumours, encephalitis, spinal cord lesions), multiple injuries, hip surgery and burns. In addition to this acquired form, hereditary causes also exist, such as fibrodysplasia ossificans progressiva, progressive osseous heteroplasia and Albright's hereditary osteodystrophy. Although these conditions are extremely rare, they can provide useful information on the physiopathology of heterotopic ossification, and thus lead to novel and causal treatment modalities. Heterotopic ossification is no trivial complication. A limitation of the range of joint motion may have serious consequences for the daily functioning of people who are already severely incapacitated because of their original lesion. Increased contractures and spasticity, pressure ulcers and increasing pain further compromise the patient's capabilities. Consequently, we feel that attention should be paid to the pathogenesis and particularly the prevention and treatment of this disorder. PMID- 16040469 TI - Summary of the WHO Collaborating Centre for Neurotrauma Task Force on Mild Traumatic Brain Injury. AB - This report aims to summarize the key findings of a recent, systematic review of the literature performed by the WHO Collaborating Centre for Neurotrauma Task Force on Mild Traumatic Brain Injury published in a supplement of the Journal of Rehabilitation Medicine. The Task Force performed a comprehensive search and critical review of the literature published between 1980 and 2002 to assemble the best evidence on the epidemiology, diagnosis, prognosis and treatment of MTBI. The Task Force identified 38,806 citations and 743 relevant studies, of which 313 (42%) were accepted on scientific merit and formed the basis of the best evidence synthesis. PMID- 16040470 TI - Walking in postpoliomyelitis syndrome: the relationships between time-scored tests, walking in daily life and perceived mobility problems. AB - OBJECTIVE: To compare walking test results with walking in daily life, and to investigate the relationships between walking tests, walking activity in daily life, and perceived mobility problems in patients with post-poliomyelitis syndrome. SUBJECTS: Twenty-four ambulant patients with post-poliomyelitis syndrome. METHODS: Walking tests were performed at self-preferred and maximal speed. Walking activity was measured with an ambulatory activity monitor. Heart rate, step cadence and walking speed in the test and in daily life were compared. Walking speed in daily life was represented by the intensity of walking. Perceived mobility problems were assessed with the Nottingham Health Profile. RESULTS: Heart rate during walking was lower in the test at self-preferred speed than in daily life (mean difference: 11.3+/-10.4; p=0.001). Self-preferred walking speed in the test and in daily life correlated significantly (r=0.55; p=0.04). In a sub-group with a test performance below the median value, test performance correlated significantly with walking activity. No significant correlation was found between perceived mobility problems and walking activity. CONCLUSION: Walking in daily life may be more demanding than walking under standardized conditions. Patients with post-poliomyelitis syndrome with the lowest test performance walked less in daily life. Patients do not necessarily match their activity pattern to their perceived mobility problems. PMID- 16040471 TI - Cognitive functioning in post-polio patients with and without general fatigue. AB - OBJECTIVE AND DESIGN: This study examined 2 main hypotheses. First, whether patients with post-polio suffering from general fatigue (n=10) demonstrate cognitive deficits compared with patients with post-polio without general fatigue (n=10). Secondly, by systematically varied test order administration we examined whether such differences varied as a function of increasing cognitive load during cognitive testing. SUBJECTS: Twenty patients diagnosed with post-polio syndrome, 10 with general fatigue and 10 without fatigue. RESULTS: Neither of the 2 hypotheses were confirmed. The group with general fatigue reported elevated levels of depression. However, no systematic association between level of depression and cognitive performance could be detected. CONCLUSION: The results of this study provide no evidence that general fatigue or cognitive load affects cognitive functioning in post-polio. PMID- 16040472 TI - Psychosocial differences as predictors for recovery from chronic low back pain following manipulation, stabilizing exercises and physician consultation or physician consultation alone. AB - OBJECTIVE: Three psychosocial profile groups are introduced in the Multidimensional Pain Inventory for chronic pain patients. Patients with the dysfunctional profile have shown a more favourable outcome after multidisciplinary treatments, due to the suggested effects of specific psychosocial treatment elements. In this study we explored, among patients with chronic low back pain, whether the Multidimensional Pain Inventory patient profile groups might respond differently to treatment without planned psychosocial elements. METHODS: Of 204 voluntarily recruited patients with chronic low back pain, 102 were randomized to a combined manipulation, exercise and physician consultation group (called the combination group) and 102 to a consultation-alone group. RESULTS: Although all subjects showed improvement during follow-up both on the Oswestry index and the Visual Analogue Scale, the dysfunctional profile patients in the combination group improved the most. Their high pre-treatment ratings on Oswestry and Visual Analogue-scales fell at the 5- and 12-month follow-ups to the same level as those of the adaptive copers or interpersonally distressed patients, and they were on a significantly lower level than the dysfunctional profile patients in consultation group during follow-up. All dysfunctional profile patients also showed a decrease in affective distress, equally in combination and consultation groups. CONCLUSION: We suggest that dysfunctional profile patients are more sensitive to respond even to treatment without any specific psychosocial elements. This should be considered when evaluating any treatment effects. Among dysfunctional profile patients, pain related anxiety and decreased acceptance of pain may contribute to their sensitivity to treatment. PMID- 16040473 TI - Recent-onset rheumatoid arthritis: a 1-year observational study of correlations between health-related quality of life and clinical/laboratory data. AB - OBJECTIVE: To analyse correlations within and between clinical/laboratory assessments and health-related quality of life variables for recent-onset rheumatoid arthritis at the time of diagnosis and 12 months later. METHODS: A total of 297 patients with recent-onset (< or =12 months) rheumatoid arthritis were included at diagnosis and followed up for 12 months. Clinical/laboratory assessment was performed by erythrocyte sedimentation rate, C-reactive protein, 28-joint count of tender/swollen joints, physician's global assessment, grip force, grip ability, functional impairment and walking speed. The self-reported health-related quality of life included symptoms (pain, morning stiffness), patients estimated general health, Health Assessment Questionnaire and SF-36. RESULTS: All tested variables improved within 6 months of diagnosis and then remained stable but still affected at the 12-month follow-up. Multivariate correlations between clinical/laboratory variables and health-related quality of life were weak. At inclusion, clinical/laboratory assessments explained 18% of health-related quality of life at the same time-point and predicted 7% of the variation in health-related quality of life after 12 months. CONCLUSION: The time course followed similar patterns for most variables, but only a small part of the variation in health-related quality of life was explained or predicted by the clinical/laboratory variables. This implies that health-related quality of life adds important information to clinical/laboratory assessments in clinical practice and should be considered in goal setting together with clinical/laboratory assessment in order to optimize healthcare and outcome. PMID- 16040474 TI - Assessment of capacity for myoelectric control: a new Rasch-built measure of prosthetic hand control. AB - OBJECTIVE: To report the results from a Rasch rating scale analysis of the Assessment of Capacity for Myoelectric Control (ACMC) implemented to evaluate internal scale validity, person response validity, separation reliability, targeting and responsiveness of the measures over time. DESIGN: Longitudinal data (18 months) from a prospective study of development of capacity for myoelectric control in children and adults were used for the analysis. PATIENTS: A consecutive sample of 75 subjects (43 males, 32 females) with upper limb reduction deficiency or amputation and myoelectric prosthetic hands referred for occupational therapy from September 2000 to March 2002. Participants' ages ranged from 2 to 57 years. METHODS: Outcome measure was the ACMC. Occupational therapists completed 210 assessments at an arm prosthesis centre in Sweden. A two faceted rating scale analysis of the data was performed. RESULTS: All 30 ACMC items and 96.2% of participants demonstrated goodness-of-fit to the rating scale model for the ACMC. Separation and SE values suggested adequate reliability of the item and person estimates. CONCLUSION: The items demonstrated internal scale validity and the participants demonstrated person response validity. The ACMC was well targeted and sensitive enough to detect expected change in ability. PMID- 16040475 TI - Influence of modified work on return to work for employees on sick leave due to musculoskeletal complaints. AB - OBJECTIVE: To determine which individual and work-related factors are associated with performing modified work and to evaluate the influence of modified work on the duration of sick leave and health-related outcomes among employees with musculoskeletal complaints. STUDY DESIGN: A prospective study with 12 months follow-up. METHODS: In this prospective study a total of 164 employees on sick leave for 2-6 weeks due to musculoskeletal complaints completed 2 questionnaires. At baseline we gathered information about individual characteristics, physical and psychosocial workload, and disease specific and general health. The follow-up questionnaire, sent to respondents who returned to their original job on full duty, collected information about having performed modified work, and disease specific and general health. RESULTS: Employees were less likely to perform modified work when their regular work was characterized by frequent lifting and their relationship with colleagues was less than good. Employees were more likely to return to modified work when they had a better mental health, had prolonged periods of standing in their regular job and had less skill discretion. Duration of sick leave was influenced by chronicity of complaints and disability, but not by modified work. CONCLUSION: Modified work, as the only advice given by a occupational health physician, did not influence the total duration of sick leave nor the improvement in health during sick leave for employees on sick leave due to musculoskeletal complaints. PMID- 16040476 TI - Impairments, activity limitations and participation restrictions 6 and 12 months after breast cancer operation. AB - OBJECTIVE: To describe the impairments of upper body and limbs, activity limitations and participation restrictions 6 and 12 months after operation for breast cancer and to examine the impact of impairments on activity limitations. DESIGN: A prospective survey 6 and 12 months after operation. PATIENTS: Ninety six breast cancer patients. METHODS: A questionnaire for assessing the impairments, activity limitations and participation restrictions was developed. RESULTS: The most common impairments 6 months after operation were breast and axilla scar tightness, axilla oedema and neck-shoulder pain. At 12-month follow up the breast scar tightness (p=0.008) and axilla oedema (p=0.023) decreased, and limb ache (p=0.005) increased significantly. The most limiting impairments were axilla oedema and limb numbness 6 months after operation, and at 12-month follow up axilla oedema. Lifting, carrying and reaching out caused worsening of impairments to more than half of the respondents at 6-month follow-up. Regression analysis showed that many impairments together were determinants of activity limitations and sleep impairment. Participation restrictions were constant. Respondents had not given up participation in activities in the home, but some had abandoned leisure activities and felt that their work ability had decreased. CONCLUSION: Impairments and their impact on activities were frequent and constant. There is an urgent need for developing rehabilitation protocols for breast cancer patients. PMID- 16040477 TI - Comparison of disordered swallowing patterns in patients with recurrent cortical/subcortical stroke and first-time brainstem stroke. AB - OBJECTIVE: To describe the disordered swallowing patterns in recurrent cortical/subcortical stroke and first-time brainstem stroke. DESIGN: A retrospective study. SUBJECTS: Forty-seven consecutive patients, 28 with recurrent cortical/subcortical stroke and 19 with first-time brainstem stroke, referred for dysphagic evaluation to the rehabilitation department of a medical centre. METHODS: Thirty-five male and 12 female patients with a mean age of 62.0+/-11.5 years were included. The median post-stroke duration was 17.0 days. The records of clinical examination and a videofluoroscopic study of swallowing were collected through chart review. The percentages of abnormalities seen at clinical examination and videofluoroscopic swallowing study between recurrent cortical/subcortical stroke and first-time brainstem stroke patients were compared using a chi-square test. RESULTS: The recurrent cortical/subcortical patients suffered from a higher rate of impaired tongue movement, drooling and aphasia at clinical examination and a higher percentage of swallowing abnormalities in oral-preparatory and oral phases in the videofluoroscopic swallowing study. The abnormal videofluoroscopic findings in first-time brainstem stroke patients predominantly occurred in the pharyngeal phase. Both groups had more difficulties swallowing thin barium than they did swallowing the thick and paste barium. CONCLUSION: The recurrent cortical/subcortical stroke and first time brainstem stroke patients show different manifestations in some parameters of both clinical examination and videofluoroscopic swallowing study. PMID- 16040518 TI - Ergonomics in the digital age. Proceedings of the XVth Triennial Congress of the International Ergonomics Association and 7th Joint Conference of the Ergonomics Society of Korea/Japan Ergonomics Society. August 24-29, 2003. Seoul, Korea. PMID- 16040519 TI - Ergonomics and human factors: the paradigms for science, engineering, design, technology and management of human-compatible systems. AB - This paper provides a theoretical perspective on human factors and ergonomics (HFE), defined as a unique and independent discipline that focuses on the nature of human-artefact interactions, viewed from the unified perspective of the science, engineering, design, technology and management of human-compatible systems. Such systems include a variety of natural and artificial products, processes and living environments. The distinguishing features of the contemporary HFE discipline and profession are discussed and a concept of ergonomics literacy is proposed. An axiomatic approach to ergonomics design and a universal measure of system-human incompatibility are also introduced. It is concluded that the main focus of the HFE discipline in the 21st century will be the design and management of systems that satisfy human compatibility requirements. PMID- 16040520 TI - The future of research in understanding and controlling work-related low back disorders. AB - Our knowledge of low back disorder (LBD) causation has progressed well over the years with in-depth understanding accelerating in the traditional disciplines of biomechanics, psychology, psychophysics, psychosocial, physiology, genetics, organizational psychology and rehabilitation. However, each of these disciplines has studied LBD causality in isolation of other disciplines. The underlying assumption is that each discipline can fully explain causality and each discipline is treated as if it were mutually exclusive and exhaustive of the other disciplines. Hence, the body of knowledge has progressed along research silos where we have in-depth knowledge along given research tracks that are defined by the boundaries of the discipline. Furthermore, a wealth of knowledge has been amassed within each of these research silos. How can they all be correct if they are indeed mutually exclusive and exhaustive? The answer is: they cannot be. This brief review of the state-of-the art in LBD research applied to ergonomics, suggests that instead of observing LBD through the myopic lens of each discipline, we need to begin to view LBD causality as a system. Recent work attempting to understand the interaction between these traditional disciplines has demonstrated that many of the findings along these silos are really interrelated and can be explained in terms of changes in the biomechanical loading at the tissue level. It is argued that further efforts to understand these interactions represent the next level of understanding causality of LBDs. PMID- 16040521 TI - Improving digital human modelling for proactive ergonomics in design. AB - This paper presents the need to improve existing digital human models (DHMs) so they are better able to serve as effective ergonomics analysis and design tools. Existing DHMs are meant to be used by a designer early in a product development process when attempting to improve the physical design of vehicle interiors and manufacturing workplaces. The emphasis in this paper is placed on developing future DHMs that include valid posture and motion prediction models for various populations. It is argued that existing posture and motion prediction models now used in DHMs must be changed to become based on real motion data to assure validity for complex dynamic task simulations. It is further speculated that if valid human posture and motion prediction models are developed and used, these can be combined with psychophysical and biomechanical models to provide a much greater understanding of dynamic human performance and population specific limitations and that these new DHM models will ultimately provide a powerful ergonomics design tool. PMID- 16040522 TI - Quantitative postural load assessment for whole body manual tasks based on perceived discomfort. AB - Many Korean workers are exposed to repetitive manual tasks or prolonged poor working postures that are closely related to back pain or symptoms of musculoskeletal disorders. Workers engage in tasks that require not only handling of heavy materials, but also assuming prolonged or repetitive non-neutral work postures. Poor work postures that have been frequently observed in the workplaces of shipbuilding shops, manufacturing plants, automobile assembly lines and farms often require prolonged squatting, repetitive arm raising and wrist flexion and simultaneous trunk flexion and lateral bending. In most manufacturing industries, workers have to assume improper work postures repetitively, several hundreds of times per day depending on daily production rate. A series of psychophysical laboratory experiments were conducted to evaluate the postural load at various joints. A postural load assessment system was then developed based on a macro postural classification scheme. The classification scheme was constructed based on perceived discomfort for various joint motions as well as previous research outcomes. On the basis of the perceived discomfort, postural stress levels for the postures at individual joints were also defined by a ratio scale to the standing neutral posture. Laboratory experiments simulating automobile assembly tasks were carried out to investigate the relationship between body-joint and whole-body discomfort. Results showed a linear relationship between the two types of discomfort, with the shoulder and low back postures being the dominant factor in determining the whole body postural stresses. The proposed method was implemented into a computer software program in order to automate the procedure of analysing postural load and to enhance usability and practical applicability. PMID- 16040523 TI - Optimizing the interaction of children with information and communication technologies. AB - This paper outlines the major changes in the lives of children in industrially advanced countries associated with the increased interaction with information and communication technologies. The potential opportunities and threats to the cognitive, social, physical and visual development of children are reviewed to emphasize the importance of optimizing the interaction. The change in children's use of technology also poses opportunities and threats for ergonomics that should be noted if the profession is to continue being relevant and useful into this century. The paper ends with a pathway to the development and implementation of guidelines about child information and communication technology use for different groups of guideline users. PMID- 16040524 TI - Providing cognitive support with technology-based warning systems. AB - New and emerging technologies promise to revolutionize risk communication. The benefits of technology are discussed with regard to the components of a recently described warning process model. Examples are provided to illustrate how technology can potentially improve information accessibility and cognitive support. Design principles such as warning interactivity, dynamic modification and personalization are considered as potential applications of technology that should enhance warning effectiveness in future technology-based systems. PMID- 16040525 TI - From industrial safety to corporate health management. AB - Occupational health and safety is one of the most important topics of ergonomics. In many countries the practical issues are addressed in worker protection laws or rules and most companies have some sort of occupational health and safety system. As with many other ergonomics approaches, such activities are often necessary because of legal requirements. Such efforts are often viewed by management as primarily cost intensive. To change this image, it would be helpful to have a more positive, management-oriented approach. Corporate health management as a module of an integrated management system can fulfil this goal. PMID- 16040526 TI - Ergonomics in total quality management: how can we sell ergonomics to management? AB - The objective of this paper is to describe how and why ergonomics should be promoted in total quality management (TQM). Ergonomics and TQM activities are compared. An approach is proposed to apply ergonomics in TQM using ergonomics circles. An eight-step approach is introduced for applying ergonomics using ergonomics circles and a study that employed this approach in Korea is discussed. In applying this approach, all processes were first evaluated by workers. Processes that were identified as problematic were analysed by a company-wide committee to set priorities for improvement. An ergonomics improvement team consisting of safety and health personnel, process engineers and management innovation personnel then worked on the processes using a low-cost approach. It was found that applying ergonomics using ergonomics circles as quality circles in TQM was effective in improving workplaces and resulted in increasing productivity, cost saving and improved safety. PMID- 16040527 TI - Enhancing the impact of ergonomics interventions. AB - The aim of this paper was to provide recommendations for ergonomists involved in project design and evaluation. Twelve research and consultancy projects involving ergonomics interventions in a variety of settings in The Netherlands were analysed to identify factors important to success. Projects included participatory and non-participatory cases. Eight factors were identified for successful project design and for demonstrating project effectiveness. PMID- 16040528 TI - Ergonomics support for local initiative in improving safety and health at work: International Labour Organization experiences in industrially developing countries. AB - Ergonomics has played essential roles in the technical cooperation activities of the International Labour Organization (ILO) in occupational safety and health in industrially developing countries. Ergonomics support focusing on practical day to-day needs at the grass-root workplace has strengthened the local initiative in improving safety and health. Practical action-tools such as ergonomics checklists, local good example photos and group discussions have assisted workers and employers in identifying feasible solutions using locally available resources. Direct participation of workers and employers has been promoted in ergonomics training aimed at immediate solutions. ILO Guidelines on Occupational Safety and Health Management Systems have played increasingly important roles in the systematic planning of local improvement actions. Policy-level programmes to develop network support mechanisms to the grass-root workplace were essential for following up and sustaining local achievements. Practical ergonomics support tools, such as action checklists and low-cost improvement guides, should be developed and widely applied so as to reach grass-root levels and help local people create safer and healthier workplaces. PMID- 16040530 TI - High scores on the Western Aphasia Battery correlate with good functional communication skills (as measured with the Communicative Effectiveness Index) in aphasic stroke patients. AB - OBJECTIVE: To examine the correlation between an impairment-level and a functional-level assessment scale of aphasia. DESIGN: Prospective, longitudinal study. SETTING: A stroke rehabilitation unit. SUBJECTS: Sixty-seven aphasic acute stroke patients who were undergoing a multi-disciplinary rehabilitation programme, including conventional speech and language therapy (SLT). INTERVENTION: Patients were assessed on study entry and 4, 8, 12 and 24 weeks after the start of SLT. The language impairment was assessed with the Western Aphasia Battery (WAB) and the communicative functional limitation associated with aphasia was measured with the Communicative Effectiveness Index (CETI).Results. There was a statistically significant correlation between the two scales for all assessment periods (Pearson's r=0.71; P<0.01). CONCLUSION: The study suggests that in the acute and subacute stages of stroke the scores of WAB and CETI can be surmised from one another. PMID- 16040531 TI - Reactions to invisible disability: the experiences of young women survivors of hemorrhagic stroke. AB - PURPOSE: To discuss how young, female and invisibly disabled, long-term survivors of hemorrhagic stroke experience the reactions of others as they negotiate the social environment. METHOD: Open-ended and in-depth interviews were conducted with 22 women to learn about their post-stroke experiences, and the interviews were analyzed for common issues and themes. RESULTS: Participants expressed concerns about the reactions of others in the context of discussing popular understandings about who is affected by stroke, and the significance of having invisible disabilities. Participants' experiences were mediated by the cultural belief that stroke is a disease of old age, and by the belief that disabilities worth taking seriously are readily visible. The existence of these beliefs about stroke and disability made it difficult for participants to deal with the reactions of others. CONCLUSIONS: Participants must negotiate their everyday lives within a social context that they are ill-prepared to deal with. Rehabilitation practices need to take this into account and counsel stroke survivors about what to expect and what they need to do for a good QOL in the community. PMID- 16040532 TI - Long-term survival following traumatic brain injury. AB - PURPOSE: The study used a retrospective cohort design to establish long-term mortality rates and predictors of mortality for persons after moderate to severe traumatic brain injury (TBI). METHOD: Consecutive records of persons with moderate to severe TBI who were discharged from a large rehabilitation hospital in Pittsburgh, Pennsylvania in the years 1974-1984, 1988 and 1989 were reviewed. RESULTS: Six hundred and forty-two eligible individuals were identified and mortality was ascertained up to 24 years post injury. One hundred and twenty eight of these individuals were found to be deceased. Poisson regression analyses revealed at least a 2-fold increased risk for mortality compared to the general population. Pre-injury characteristics and levels of disability at discharge from in-patient rehabilitation were among the strongest predictors of mortality. CONCLUSIONS: These data constitute evidence for premature death in the post-acute TBI population following a moderate to severe head injury and are discussed in relation to other research in the area. PMID- 16040533 TI - Issues for selection of outcome measures in stroke rehabilitation: ICF activity. AB - PURPOSE: To evaluate the psychometric and administrative properties of outcome measures in the WHO International Classification of Functioning, Disability and Health (ICF) Activity category used in stroke rehabilitation research and reported in the published literature. METHOD: Critical review and synthesis of measurement properties for nine commonly reported instruments in the stroke rehabilitation literature. Each instrument was rated using the eight evaluation criteria proposed by the UK Health Technology Assessment (HTA) programme. The instruments were also assessed for the rigour with which their reliability, validity and responsiveness were reported in the published literature. RESULTS: The reporting of specific measurement qualities for outcome instruments was relatively consistent across measures located within the same general ICF category. There was evidence to suggest that the measures were responsive to change as well as being valid and reliable tools. The best available instruments were associated with the assessment of activities of daily living, balance (static and dynamic), functional independence, and functional mobility. CONCLUSIONS: Given the diversity that exists among available measures, the reader is encouraged to examine carefully the nature and scope of outcome measurement used in reporting the strength of evidence for improved functional activity in stroke rehabilitation. However, there appears to be good consensus regarding the most important indicators of successful rehabilitation outcome, especially in the case of functional mobility. PMID- 16040534 TI - Duration and intensity of physical activity and disability among European elderly men. AB - PURPOSE: To investigate the relationship between duration and intensity of physical activity and disability 10 years later, and to investigate the possible effect of selective mortality. METHOD: Longitudinal data of 560 men aged 70?-?89 years, without disability at baseline from the Finland, Italy and The Netherlands Elderly (FINE) Study was used. Physical activity in 1990 was based on activities like walking, bicycling and gardening. Disability severity (three categories) in 1990 and 2000 was based on instrumental activities, mobility and basic activities of daily living. RESULTS: Men in the highest tertile of total physical activity had a lower risk of disability than men in the lowest tertile (odds ratio (OR) 0.46; 95% confidence interval (CI): 0.26-0.84). This was due to duration of physical activity (OR highest tertile 0.42; 95% CI: 0.23-0.78 compared to the lowest tertile). Intensity of physical activity was not associated with disability. Addition of deceased men as fourth category leaded to weaker associations between physical activity and disability (OR highest tertile 0.67; 95% CI: 0.44-1.02). CONCLUSIONS: Even in old age among relatively healthy men, a physically active lifestyle was inversely related to disability. To prevent disability duration of physical activity seems to be more important than intensity. PMID- 16040535 TI - Applying the ICF for the acute hospital and early post-acute rehabilitation facilities. PMID- 16040536 TI - Rationale and principles of early rehabilitation care after an acute injury or illness. AB - Patients hospitalized for an acute illness or injury are at risk of experiencing a significant loss of functioning as defined by the International Classification of Functioning, Disability and Health (ICF). The risk of a significant loss of functioning is increased in critically ill patients, in patients with complications or long-term intensive care stays, in persons with disabilities or with pre-existing chronic conditions and in the elderly. Early identification of rehabilitation needs and early start of rehabilitation can reduce healthcare costs by reducing dependence and nursing care, length of stay and prevention of disability. Two principles of rehabilitation for acute and early post-acute care can be distinguished. First, the provision of rehabilitation by health professionals who are generally not specialized in rehabilitation in the acute hospital. And second, specialized rehabilitation care provided by an interdisciplinary team. There is large variation how this specialized, typically post-acute rehabilitation care is organized, provided, and reimbursed in different countries, regions, and settings. For instance, it may be provided either in the acute hospital or in a rehabilitation or nursing setting. Most in patients do not receive specialized rehabilitation at all during their whole stay in the acute hospital. But, it is important to point out that health professionals working in acute hospitals and who are not specialized in rehabilitation need to be able to recognize patients' needs for rehabilitation care and to perform rehabilitation interventions themselves or to assign patients to appropriate rehabilitation care settings. The principles outlined in this paper can serve as a basis for the development of clinical assessment instruments to describe and classify functioning, health and disability of patients receiving acute or early post-acute rehabilitation care. PMID- 16040537 TI - ICF Core Sets development for the acute hospital and early post-acute rehabilitation facilities. AB - The goal of this paper is to report on the background and the methods used in the ICF Core Set development for patients in the acute hospital and early post-acute rehabilitation facilities. ICF Core Sets are sets of categories out of the International Classification of Functioning, Disability and Health (ICF) which can serve as minimal standards for the assessment, communication and reporting of functioning and health for clinical studies, clinical encounters and multi professional comprehensive assessment and management. The ICF Core Sets were developed in a formal decision-making and consensus process, integrating evidence gathered from preliminary studies and expert opinion. The Acute ICF Core Sets for patients with neurological, musculoskeletal and cardiopulmonary conditions are intended for use by physicians, nurses, therapists and other health professionals working in the acute hospital on medical, surgical or other units not specialised in rehabilitation. The Post-acute ICF Core Sets for geriatric patients and patients with neurological, musculoskeletal or cardiopulmonary conditions are intended for use by physicians, nurses, therapists and other health professionals involved in early post-acute rehabilitation. The Acute and Post-acute ICF Core Sets are first versions and need to be tested and validated in the patient and professional perspective and in different countries, regions, health care and provider settings. PMID- 16040538 TI - ICF Core Set for patients with neurological conditions in the acute hospital. AB - PURPOSE: The aim of this consensus process was to decide on a first version of the ICF Core Set for patients with neurological conditions in the acute hospital. METHODS: The ICF Core Set development involved a formal decision-making and consensus process, integrating evidence gathered from preliminary studies including focus groups of health professionals, a systematic review of the literature and empiric data collection from patients. RESULTS: Twenty-one experts selected a total of 85 second-level categories. The largest number of categories was selected from the ICF component Body Functions (41 categories or 48%). Five (6%) of the categories were selected from the component Body Structures, 18 (21%) from Activities and Participation and 21 (25%) from Environmental Factors. CONCLUSIONS: The Acute ICF Core Set for patients with neurological conditions provides all professionals with a clinical framework to comprehensively assess patients in the acute hospital. This first ICF Core Set will be further tested through empiric studies in German-speaking countries and internationally. PMID- 16040539 TI - ICF Core Set for patients with cardiopulmonary conditions in the acute hospital. AB - PURPOSE: The aim of this consensus process was to decide on a first version of the ICF Core Set for patients with cardiopulmonary conditions in the acute hospital. METHODS: The ICF Core Set development involved a formal decision-making and consensus process, integrating evidence gathered from preliminary studies including focus groups of health professionals, a systematic review of the literature and empiric data collection from patients. RESULTS: Twenty-two experts selected a total of 48 second-level categories. The largest number of categories was selected from the ICF component Body Functions (21 categories or 44%). Four (8%) of the categories were selected from the component Body Structures, 10 (21%) from the component Activities and Participation, and 13 (27%) from the component Environmental Factors. CONCLUSION: The Acute ICF Core Set for patients with cardiopulmonary conditions provides all professionals with a clinical framework to comprehensively assess patients in the acute hospital. This first ICF Core Set will be further tested through empiric studies in German-speaking countries and internationally. PMID- 16040540 TI - ICF Core Set for patients with musculoskeletal conditions in the acute hospital. AB - PURPOSE: The aim of this consensus process was to decide on a first version of the ICF Core Set for patients with musculoskeletal conditions in the acute hospital. METHODS: The ICF Core Set development involved a formal decision-making and consensus process integrating evidence gathered from preliminary studies including focus groups of health professionals, a systematic review of the literature and empiric data collection from patients. RESULTS: Twenty-one experts selected a total of 47 second-level ICF categories. The largest number of categories was selected from the ICF component Body Functions (17 categories or 36%). Nine (19%) of the categories were selected from the component Body Structures, 11 (23%) from the component Activities and Participation, and 10 (21%) from the component Environmental Factors. CONCLUSION: The Acute ICF Core Set for patients with musculoskeletal conditions provides all professionals with a clinical framework to comprehensively assess patients in the acute hospital. This first ICF Core Set will be further tested through empiric studies in German speaking countries and internationally. PMID- 16040541 TI - ICF Core Set for patients with neurological conditions in early post-acute rehabilitation facilities. AB - PURPOSE: The aim of this consensus process was to decide on a first version of the ICF Core Set for neurological patients in early post-acute rehabilitation facilities. METHODS: The ICF Core Set development involved a formal decision making and consensus process, integrating evidence gathered from preliminary studies including focus groups of health professionals, a systematic review of the literature and empiric data collection from patients. RESULTS: Seventeen experts selected a total of 116 second-level categories. The largest number of categories was selected from the ICF component Body Functions (54 categories or 47%). Eleven (9%) of the categories were selected from the component Body Structures, 34 (29%) were of the categories from the component Activities and Participation, and 17 (15%) from the component Environmental Factors. CONCLUSION: The Post-acute ICF Core Set for neurological patients is a clinical framework to comprehensively assess patients in early post-acute rehabilitation facilities, particularly in an interdisciplinary setting. This first ICF Core Set will be further tested through empiric studies in German-speaking countries and internationally. PMID- 16040542 TI - ICF Core Set for patients with cardiopulmonary conditions in early post-acute rehabilitation facilities. AB - PURPOSE: The aim of this consensus process was to decide on a first version of the ICF Core Set for patients with cardiopulmonary conditions in early post-acute rehabilitation facilities. METHODS: The ICF Core Set development involved a formal decision-making and consensus process integrating evidence gathered from preliminary studies including focus groups of health professionals, a systematic review of the literature and empiric data collection from patients. RESULTS: Seventeen experts selected a total of 84 second-level categories. The largest number of categories was selected from the ICF component Body Functions (33 categories or 39% of all ICF Core Set categories). Four (5%) of the categories were selected from the component Body Structures, 23 (27%) from the component Activities and Participation, and 24 (29%) from the component Environmental Factors. CONCLUSIONS: The Post-acute ICF Core Set for patients with cardiopulmonary conditions is a clinical framework to comprehensively assess patients in early post-acute rehabilitation facilities, particularly in an interdisciplinary setting. This first ICF Core Set will be further tested through empiric studies in German-speaking countries and internationally. PMID- 16040543 TI - ICF Core Set for patients with musculoskeletal conditions in early post-acute rehabilitation facilities. AB - PURPOSE: The aim of this consensus process was to decide on a first version of the ICF Core Set for patients with musculoskeletal conditions in early post-acute rehabilitation facilities. METHODS: The ICF Core Set development involved a formal decision-making and consensus process integrating evidence gathered from preliminary studies including focus groups of health professionals, a systematic review of the literature and empiric data collection from patients. RESULTS: Fifteen experts selected a total of 70 second-level categories. The largest number of categories was selected from the ICF component Body Functions (23 categories or 33%). Seven (10%) of the categories were selected from the component Body Structures, 22 (31%) from the component Activities and Participation, and 18 (26%) from the component Environmental Factors. CONCLUSION: The Post-acute ICF Core Set for patients with musculoskeletal conditions is a clinical framework to comprehensively assess patients in early post-acute rehabilitation facilities, particularly in an interdisciplinary setting. This first ICF Core Set will be further tested through empiric studies in German speaking countries and internationally. PMID- 16040544 TI - ICF Core Set for geriatric patients in early post-acute rehabilitation facilities. AB - PURPOSE: The aim of this consensus process was to decide on a first version of the ICF Core Set for geriatric patients in early post-acute rehabilitation facilities. METHODS: The ICF Core Set development involved a formal decision making and consensus process, integrating evidence gathered from preliminary studies including focus groups of health professionals, a systematic review of the literature and empiric data collection from patients. RESULTS: Fifteen experts selected a total of 123 second-level categories. The largest number of categories was selected from the ICF component Body Functions (51 categories or 41%). 14 (11%) of the categories were selected from the component Body Structures, 30 (29%) from the component Activities and Participation, and 28 (23%) from the component Environmental Factors. CONCLUSION: The Post-acute ICF Core Set for geriatric patients is a clinical framework to comprehensively assess patients in early post-acute rehabilitation facilities, particularly in an interdisciplinary setting. This first version of the ICF Core Set will be further tested through empiric studies in German-speaking countries and internationally. PMID- 16040545 TI - Systematic review of measures and their concepts used in published studies focusing on rehabilitation in the acute hospital and in early post-acute rehabilitation facilities. AB - PURPOSE: To identify outcome measures cited in published studies focusing on rehabilitation in the acute hospital and in early post-acute rehabilitation facilities, and to identify and quantify the concepts contained in these measures using the ICF as a reference. METHODS: Electronic searches of Medline, Embase, CINAHL, Pedro and the Cochrane Library from 1997 to March 2002 were carried out. In a first step, abstracts of the retrieved studies were checked and data on the outcome measures and certain characteristics of the included studies were extracted. In a second step, the items of the questionnaires and their underlying concepts were specified. These concepts were then linked to ICF categories using standardized linkage rules. RESULTS: From the 1,657 abstracts retrieved, 259 studies met the inclusion criteria. In a second step, 277 formal assessment instruments and 351 single clinical measures were retrieved. A total of 1,353 concepts were extracted from the clinical and technical measures. Ninety-six percent of these concepts could be linked to ICF categories. Fifty-six second level ICF categories representing the concepts contained in the measures. Twenty six (46%) of the 56 categories belong to the component Body Functions, five (9%) to the component Body Structures, and 25 (45%) to the component Activities and Participation. CONCLUSIONS: The ICF provides a valuable reference to identify and quantify the concepts of outcome measures focusing on rehabilitation in the acute hospital and in early post-acute rehabilitation facilities. Our findings indicate a need to define and to agree on 'what should be measured' in rehabilitation care to allow for a comparison of patient populations. PMID- 16040546 TI - The ICF categories identified in nursing interventions administered to neurological patients with post-acute rehabilitation needs. AB - PURPOSE: The objectives of this study were (1) to determine whether the ICF covers the goals of nursing interventions and (2) to identify the areas of functioning, disability and health most relevant to nursing practice of neurological patients with early post-acute rehabilitation needs. METHODS: This cross-sectional study on nursing interventions is part of a larger multicentric cross-sectional study describing functioning, disability and health. The nursing interventions were recorded by nurses in charge of the patient. According to established rules, the recorded nursing interventions were linked to the ICF. RESULTS: One hundred and eighteen different nursing interventions were documented by the nursing staff. The intervention goals were linked to 67 different second level ICF categories. Thirty belong to the component Body Functions, 19 to the component Body Structures, 13 to the component Activities and Participation, and five to the component Environmental Factors. CONCLUSIONS: The fact that all nursing interventions in early post-acute rehabilitation facilities could be linked to ICF categories supports the usefulness of the ICF in nursing. Initial problems in adapting profession-specific terminology to the ICF framework can be overcome, as shown by our linking exercise. Further analyses will be needed to show if these theoretical findings are supported by empirical evidence. PMID- 16040547 TI - Identification of relevant ICF categories by health professionals in the acute hospital. AB - PURPOSE: To identify the most relevant problems of patients in the acute hospital in the view of health professionals using focus groups followed by a Delphi process. METHODS: Focus group and Delphi methodology were applied. The focus groups were conducted at three university hospitals. Three groups focusing on the problems in patients with neurological/neurosurgical, cardiopulmonary and musculoskeletal conditions were conducted at each hospital. The participants had to decide about which categories of the International Classification of Functioning, Disability and Health are relevant to patients in the acute hospital. The results from the focus groups were then followed by a Delphi process. RESULTS: Forty health professionals participated in the nine focus groups, 14 in the three neurological/neurosurgical groups, 13 in the three musculoskeletal groups, 13 in the three cardiopulmonary groups. One hundred and sixty-four of the second-level ICF categories (61.7% of all second-level categories) were considered as relevant by the health professionals of the neurological group, 100 categories (37.6%) by the musculoskeletal group and 91 (34.2%) by the cardiopulmonary group. CONCLUSIONS: The involvement of experts from different health professions is a valuable tool to identify typical patient characteristics, expressed as distinct ICF categories, which can be used across different acute conditions. PMID- 16040548 TI - Identification of relevant ICF categories by patients in the acute hospital. AB - PURPOSE: To describe functioning and health of patients in the acute hospital and to identify the most common problems using the International Classification of Functioning, Disability and Health (ICF). METHODS: Cross-sectional survey in a convenience sample of neurological, musculoskeletal and cardiopulmonary patients requiring rehabilitation in the acute hospital. The second level categories of the ICF were used to collect information on patients' problems. For the ICF components Body Functions, Body Structures and Activities and Participation absolute and relative frequencies of impairments/limitations in the study population were reported. For the component Environmental Factors absolute and relative frequencies of perceived barriers or facilitators were reported. RESULTS: The mean age in the sample was 57.6 years with a median age of 60.5, 49% of the patients were female. In 101 patients with neurological conditions, 115 ICF categories had a prevalence of 30% and more: 32 categories of Body Functions, 13 categories of Body Structures, 32 categories of Activities and Participation and 38 categories of Environmental Factors. In 105 patients with cardiopulmonary conditions, 80 categories had a prevalence of 30% and more: 36 categories of Body Functions, eight categories of Body Structures, 10 categories of Activities and Participation and 26 categories of Environmental Factors. In 90 patients with musculoskeletal conditions, 61 categories had a prevalence of 30% and more: 14 categories of Body Functions, five categories of Body Structures, 16 categories of Activities and Participation and 26 categories of Environmental Factors. CONCLUSION: This study is a first step towards the development of ICF Core Sets for patients in the acute hospital. PMID- 16040549 TI - Identification of relevant ICF categories by patients with neurological conditions in early post-acute rehabilitation facilities. AB - PURPOSE: To describe functioning and health of patients with neurological conditions in early post-acute rehabilitation facilities and to identify the most common problems using the International Classification of Functioning, Disability and Health (ICF). METHODS: Cross-sectional survey in a convenience sample of patients with neurological conditions requiring rehabilitation in early post acute facilities. The second-level categories of the ICF were used to collect information on patients' problems. For the ICF components Body Functions, Body Structures and Activities and Participation absolute and relative frequencies of impairments/limitations in the study population were reported. For the component Environmental Factors absolute and relative frequencies of perceived barriers or facilitators were reported. RESULTS: The mean age in the sample was 56.6 years with a median age of 60 years. Forty percent of the patients were female. In 292 neurological patients 125 categories (51%) had a prevalence of 30% and above: 39 categories (49%) of Body Functions, 11 categories (28%) of Body Structures, 64 categories (88%) of Activities and Participation and 10 (20%) categories of Environmental Factors. CONCLUSION: This study is a first step towards the development of ICF Core Sets for of patients with neurological conditions in early post-acute rehabilitation facilities. PMID- 16040550 TI - Identification of relevant ICF categories by geriatric patients in an early post acute rehabilitation facility. AB - PURPOSE: To describe functioning and health of elderly patients in an early post acute rehabilitation facility and to identify the most common problems using the International Classification of Functioning, Disability and Health (ICF). METHODS: Cross-sectional survey in a convenience sample of elderly patients requiring rehabilitation in an early post-acute rehabilitation facility. The second-level categories of the ICF were used to collect information on patients' problems. For the ICF components Body Functions, Body Structures and Activities and Participation, absolute and relative frequencies of impairments/limitations in the study population were reported. For the component Environmental Factors absolute and relative frequencies of perceived barriers or facilitators were reported. RESULTS: The mean age in the sample was 79.9 years. Sixty-nine percent of the patients were female. In 150 patients, 82 ICF categories (34%) had a prevalence of 30% or above. The 82 categories included 22 categories (45%) of the component Body Functions, six categories (15%) of the component Body Structures, 25 categories (34%) of the component Activities and Participation and 29 (57%) categories of the component Environmental Factors. CONCLUSION: This study is a first step towards the development of ICF Core Sets for geriatric patients in early post-acute rehabilitation facilities. PMID- 16040551 TI - Statistical methods in epidemiology. VIII. On the use of likelihood ratios for diagnostic testing with an application to general practice. AB - PURPOSE: An array of statistical methods is available for 2 x 2 contingency table analysis. These include odds ratios, reliability coefficients and the Chi-squared test. This paper continues the '2 x 2' theme but extends it to look at diagnostic testing. The setting is primary care, the disease is urological cancer. METHOD: Several statistical indices are discussed to measure diagnostic efficiency; these include sensitivity, specificity and the likelihood ratio. The latter can be converted into a flexible 'weight of evidence', which is equivalent to the naive Bayes' discriminant. RESULTS: The utility of the naive Bayes' discriminant is illustrated in a primary care setting. CONCLUSION: The naive Bayes' discriminant has proved to be the most durable of statistical discriminants in medical diagnosis. The reason for its durability is ease of use. Despite a concern about independence, it remains a popular choice as a diagnostic aid. PMID- 16040552 TI - Work-related determinants of return to work of employees on long-term sickness absence. AB - PURPOSE: The aim of the study is to identify work-related determinants of return to work (RTW) of employees who are on long-term sickness absence. METHOD: The study was based on a sample of 926 employees on sickness absence (maximum duration of 12 weeks). The employees filled out a baseline questionnaire and were subsequently followed until the 10th month after listing sick. Cox proportional hazards regression analyses were used to identify determinants of RTW. RESULTS: Working in one of the vocational sectors public administration, construction, financial and commercial services, transport, or education (P = 0.00) and having low co-worker support (P = 0.01) were related to longer duration to RTW in the multivariate model. Having low supervisor support (P = 0.01) was associated with a higher RTW rate. CONCLUSIONS: Vocational sector is a strong predictor of RTW. Especially employees from the sector education are slow as to RTW. The observed association between low supervisor support and RTW was unexpected. However, the study confirms earlier research on the association between low co-worker support and RTW. PMID- 16040553 TI - Long duration spinal cord injury: perceptions of functional change over time. AB - PURPOSE: To investigate perceptions of functional change over time held by individuals with long duration spinal cord injury (SCI) living in Queensland, Australia. METHOD: A retrospective telephone questionnaire was administered to 84 individuals who had sustained a SCI more than 20 years previously and were older than 15 at the time of injury. Motor subset scores of the Functional Independence Measure (FIM) and a single scale measuring mobility aids status (MAIDS), were collected for three points in time--post discharge from initial rehabilitation (D/C point); approximately 10 years post injury (Mid point) and currently (Current point). RESULTS: A significant number of participants perceived that their function had increased between the D/C and Mid points and had subsequently decreased between the Mid and Current points. Participants also reported an increasing dependence on mobility aids between the Mid point and the Current point. Those who reported functional decline between the Mid and Current points were significantly older than those who did not report functional decline but did not differ in duration of injury or age at onset. CONCLUSIONS: The results support the need for services that provide assessment and intervention for functional changes throughout the life span of people with SCI. PMID- 16040554 TI - A pilot study of falls risk and vestibular dysfunction in older fallers presenting to hospital emergency departments. AB - PURPOSE: To compare falls risk in older fallers and non-fallers, with an emphasis on dizziness and signs of vestibular dysfunction. METHOD: The fallers had presented to the Emergency Department of the Royal Melbourne Hospital, Australia following a fall and were discharged directly home (n = 20) (75% female, mean age 78 years). The non-fallers were an age and gender matched group, who had not fallen in the past 12 months (n = 20). All clients received a home-based assessment, which involved a comprehensive assessment of falls risk. RESULTS: Over three-quarters of the fallers took four or more medications, had balance impairments, and used a gait aid in the community. The fallers had a significantly higher falls risk score (P < 0.001), demonstrated significantly poorer balance (P < 0.001) and walked significantly more slowly (P < 0.001) than the non-fallers. There was no significant difference between the groups in their reports of dizziness (P = 0.68), although static balance testing (CTSIB condition 5) suggested a greater degree of underlying vestibular dysfunction in the group of fallers (P < 0.001). CONCLUSION: Older people discharged home from the ED following a fall are at high risk of falling in the future and have a greater level of vestibular dysfunction based on simple clinical testing. Additional clinically applicable tests of vestibular function are required to further investigate the relationship between vestibular dysfunction and falling in older people. PMID- 16040555 TI - Issues for selection of outcome measures in stroke rehabilitation: ICF Participation. AB - PURPOSE: To evaluate the psychometric and administrative properties of outcome measures in the ICF Participation category, which are used in stroke rehabilitation research and reported in the published literature. METHOD: Critical review and synthesis of measurement properties for six commonly reported instruments in the stroke rehabilitation literature. Each instrument was rated using the eight evaluation criteria proposed by the UK Health Technology Assessment (HTA) programme. The instruments were also assessed for the rigour with which their reliability, validity and responsiveness were reported in the published literature. RESULTS: Validity has been well reported for at least half of the measures reviewed. However, methods for reporting specific measurement qualities of outcome instruments were inconsistent. Responsiveness of measures has not been well documented. Of the three ICF categories, Participation seems to be most problematic with respect to: (a) lack of consensus on the range of domains required for measurement in stroke; (b) much greater emphasis on health related quality of life, relative to subjective quality of life in general; (c) the inclusion of a mixture of measurements from all three ICF categories. CONCLUSIONS: The reader is encouraged to examine carefully the nature and scope of outcome measurement used in reporting the strength of evidence for improved participation associated with stroke rehabilitation. There is no consensus regarding the most important indicators of successful involvement in a life situation and which ones best represent the societal perspective of functioning. In particular, quality of life outcomes lack adequate conceptual frameworks to guide the process of development and validation of measures. PMID- 16040556 TI - A computerised communication aid for people with aphasia. AB - PURPOSE: To develop a portable computerised communication aid for aphasic people to support communication in everyday life. METHOD: A multidisciplinary team of aphasiologists, augmentative and alternative communication specialists, speech and language therapists and technicians developed a portable, modular system, PCAD (portable communication assistant for people with dysphasia), running on a commercially available handheld computer. The system was tested in a multiple case study. Aphasia therapy services In the UK, Portugal and The Netherlands referred 28 people with aphasia, who were considered eligible for a computerised communication aid. Participants were trained following a protocol and used the device in self-chosen real life settings. RESULTS: Six of the 28 selected aphasic patients decided not to test the device; 22 participated in the training. All 22 learned to operate the aid, 17 used it functionally, in everyday life. Five people did not use the aid outside the therapy room, although they were able to operate the aid and to use it in role play. These unsuccessful clients were younger, and tended to have a shorter duration of the aphasia. CONCLUSIONS: Carefully selected aphasic patients may benefit from a computerised communication aid, using it functionally in everyday communicative settings. PMID- 16040557 TI - Letter to the editor in response to the article of Dekkers and Soballe (Activities and impairments in the early stage of rehabilitation after Colles' fracture; Vol. 26, No. 11, June 3, 2004). PMID- 16040559 TI - Testing of fibrous particles: short-term assays and strategies. PMID- 16040563 TI - Use of genotoxicity data to support clinical trials or positive genetox findings on a candidate pharmaceutical or impurity .... now what? AB - Results from carcinogenicity studies are generally not available for drugs until the time of approval. Many people, including healthy volunteers are often exposed to pharmacologically active doses of the drug before carcinogenicity results are available. The Food and Drug Administration (FDA) Center for Drug Evaluation and Research uses results of genetic toxicology studies as a surrogate for carcinogenicity during the drug development phase (clinical trials). A number of issues are considered in deciding whether drugs that give positive results in genetic toxicology studies can be given to subjects in clinical trials. These relate to the drug indication, the target population, duration of treatment, and importance of the drug. In general, single-dose clinical studies are permitted regardless of the genetox results. In situations where a genetic toxicology assay showed a positive result, some review divisions have asked sponsors to perform a Syrian hamster embryo (SHE) cell transformation assay or a p53 carcinogenicity study prior to allowing repeat-dose clinical trials to proceed. This paper discusses alternatives to SHE cell and p53 assays when faced with a positive result in a genetic toxicology assay. In addition, this paper discusses factors to consider when setting limits for genotoxic impurities in drug substances and products. PMID- 16040564 TI - Applications of emerging technologies in toxicology and safety assessment. AB - Standardized protocols for repeat-dose toxicity studies have many advantages, including experimental redundancy (i.e., the use of more than a single experimental approach in the assessment of a given organ or tissue) and evaluation of numerous tissues to ensure detection of adverse effects, as well as the ability to develop robust historical control databases. However, traditional toxicology study designs may not adequately address questions of a mechanistic nature that might provide insights on whether particular toxicology findings in animals are relevant to humans. Such questions may be more readily answered using mechanism-based technologies such as toxicogenomics, proteomics, or metabonomics. Such newer approaches may permit, for example, the detailed assessment of the transcriptional profile differences that distinguish a normal healthy tissue from a diseased or damaged tissue. The resultant information can be used to elucidate the mechanism and accompanying biomarkers for toxicity, as well as to identify potential molecular targets for therapeutic intervention by drugs. Despite their conceptual appeal, the use of emerging technologies in toxicology is accompanied by significant challenges. For example, toxicogenomic assessments entail the generation of large amounts of bioinformatic data that must be interpretable for their full value to be realized. Also, none of these newer approaches have established uniformly acceptable quality standards (e.g., such as may be defined in interlaboratory validation studies) or a track record of achievement in guiding regulatory decisions. As a result, newer techniques, at least for the present, are more likely to be focused on mechanistic questions with compounds of known toxicity (either positive indicator compounds or "failed" pharmaceutical candidates). If the use of a nascent or emerging technology is contemplated for mechanistic studies of pharmaceutical compounds later in preregistration development, it will be crucial for toxicologists to engage their regulatory colleagues in discussions at an early stage to ensure closer alignment in thought. The successful use of emerging technologies to address toxicology issues will require a close partnership between industry and regulatory agencies. PMID- 16040565 TI - TaqMan applications in genetic and molecular toxicology. AB - Quantification of nucleic acids has become a common procedure in many toxicology laboratories. Among the technologies that accomplish this is the fluorogenic 5' nuclease assay, commonly known as TaqMan. Three TaqMan applications for genetic and molecular toxicology are presented in this article: quantification of gene expression, detection of genetic polymorphisms, and quantification of chromosomal DNA deletions. Of these, quantification of gene expression is the most widely used, and established TaqMan as a benchmark technology for nucleic acid quantification. Two additional applications, polymorphism detection and quantification of DNA deletions, demonstrate the flexibility and quantitative strengths that make TaqMan so powerful, including high precision, excellent sensitivity, and broad linear dynamic range. These and similar applications improve our ability to investigate genetic and molecular dimensions of toxicological phenomena, and have promoted the widespread use of TaqMan in toxicology departments in the pharmaceutical industry. In addition to presenting these applications, the authors discuss some of the challenges of integrating TaqMan and other new technologies into the drug development process. PMID- 16040566 TI - Understanding biology through bioinformatics. AB - During the journey from the discovery of DNA to be the source of genetic information and elucidation of double-helical nature of DNA molecule to the assembly of human genome sequence and there after, bioinformatics has become an integral part of modern biology. Bioinformatics relies substantially on significant contributions made by scientists in various fields, including but not limited to, linguistics, biology, mathematics, computer science, and statistics. There is an ever increasing amount of data to elucidate toxic mechanisms and/or adverse effects of xenobiotics in the field of toxicogenomics. Annotation in combination with various bioinformatics analytical tools can play a crucial role in the understanding of genes and proteins, and can potentially help draw meaningful conclusions from various data sources. This article attempts to present a simple overview of bioinformatics, and an effort is made to discuss annotation. PMID- 16040567 TI - Application of emerging technologies in toxicology and safety assessment: regulatory perspectives. AB - Emerging technologies applied in the regulatory field encompass a group of technologies that are used in addition to or in replacement of the standard toxicology studies conducted to support an Investigational New Drug Application (IND) or New Drug Application (NDA). The standard package includes general toxicology studies of various duration, safety pharmacology studies, genetic toxicology studies, and reproductive toxicology studies. New and emerging technologies applied to the regulation of new drugs include the use of novel biomarkers, transfected cells and transgenic animals, and the "omics" technologies (toxicogenomics, proteomics, and metabonomics). These technologies are at various stages of regulatory development and acceptance. For example, the use of transgenic animals have gained acceptance by regulatory authorities to replace a 2-year carcinogenicity assay. Alternatively, the "omics" technologies are not sufficiently advanced to achieve regulatory acceptance as replacements, although these assays have a role early in drug development and they may prove useful as supplements to standard studies. Data from these assays have been used to address specific mechanistic questions in combination with standard toxicology assays. PMID- 16040568 TI - An in vivo bioassay for detecting antiandrogens using humanized transgenic mice coexpressing the tetracycline-controlled transactivator and human CYP1B1 gene. AB - The typical strategy used in analysis of antiandrogens involves the morphological changes of a marker in castrated rats Hershberger assay for the prostate, seminal vesicle, levator ani plus bulbocavernosus muscles (LABC), Cowper's gland, and glans penis. However, there are disadvantages to this approach, such as the time required, and the results may not correspond to those in actual human exposure. To evaluate its ability for detecting antiandrogens, in vivo the dose effect of di-(2-ethylhexyl) phthalate (DEHP) and time effect of five antiandrogens, DEHP, di-n-butyl phthalate (DBP), diethyl phthalate (DEP), linuron (3-(4 dichlorophenyl)-methoxy-1-methylurea), and 2,4'-DDE (1,1-dichloro-2-(p chlorophenyl)-2-(o-chlorophenyl)ethylene), were investigated using humanized transgenic mice coexpressing tetracycline-controlled transactivator (tTA) and the human cytochrome P450 (CYP) enzyme CYP1B1 (hCYP1B1). Adult transgenic males were treated with each of the five antiandrogens, and their tTA-driven hCYP1B1 expressions analyzed by real-time polymerase chain reaction (PCR) and/or Western blot and for O-debenzylation activity. Herein, the treatments of adult males with the five antiandrogens were shown to affect the increased levels of tTA-driven hCYP1B1 expression in both dose-dependent and repeated experiments. Thus, this novel in vivo bioassay, using humanized transgenic mice, is useful for measuring antiandrogens, and is a means to a more relevant bioassay relating to actual human exposure. PMID- 16040569 TI - Determination of lead in Smilax luzonensis herbal preparations in Malaysia. AB - The DCA (Drug Control Authority) of Malaysia implemented the phase 3 registration of traditional medicines on 1 January 1992. As such, a total of 100 products in various pharmaceutical dosage forms of a herbal preparation containing Smilax luzonensis, either single or combined preparations, were analyzed for the presence of lead, using atomic absorption spectrophotometry. Results showed that 14% of the above products possessed 10.02 to 21.21 ppm of lead, and, therefore, they failed to comply with the quality requirement for traditional medicines in Malaysia ( <10 ppm). Although this study showed that 86% of the products complied with the quality requirement for traditional medicines in Malaysia pertaining to lead, they cannot be assumed safe from lead contamination because of batch-to batch inconsistency. PMID- 16040570 TI - Differential effects of mixtures of cholesterol oxidation products on bovine aortic endothelial cells and human monocytic U937 cells. AB - Cholesterol oxidation products or oxysterols are of interest due to their hypothesized role in the development of atherosclerosis. The objective of the present study was to assess the cytotoxic effects of mixtures of oxysterols: 25 hydroxycholesterol (25-OHC), 7beta-hydroxycholesterol (7beta -OHC), and cholesterol-5beta,6beta -epoxide (beta -epox) on two cell types associated with the atherosclerotic process, bovine aortic endothelial (BAE) cells and human monocytic U937 cells. Cells were exposed to 25-OHC, 7beta -OHC, or beta -epox, or equimolar mixtures (30 mu M) of 25-OHC and 7beta -OHC, 25-OHC and beta-epox, or 7beta-OHC and beta -epox for 48 h. Cell viability was assessed using the fluorescein diacetate/ethidium bromide (FDA/ EtBr) assay and nuclear morphology following staining with Hoechst 33342. 25-OHC was the least toxic of the oxysterols and did not induce apoptosis in either cell line. Both 7beta-OHC and beta -epox treatments were cytotoxic and induced apoptosis in the cells. Cotreatment with 25-OHC did not alter the toxicity of 7beta -OHC and beta -epox in U937 cells but did decrease the percentage apoptotic cell death. In contrast, in the BAE cells cotreatment with 25-OHC had a slight protective effect on 7beta OHC and beta-epox-induced toxicities and a marked decrease in apoptotic cell death. The 7beta -OHC and beta -epox mixture induced a significant increase in apoptotic cell death in U937 cells but decreased this mode of cell death in the BAE cells. The effects of oxysterols on glutathione levels also differed between the cells with changes noted in U937 and not in BAE cells. Results demonstrate interactive effects when oxysterols are studied as mixtures rather than single compounds in vitro. PMID- 16040571 TI - Efficacy of chlorine dioxide as a gas and in solution in the inactivation of two trichothecene mycotoxins. AB - The efficacy of chlorine dioxide (ClO2) in detoxifying two potential bioterrorism agents, the trichothecene mycotoxins verrucarin A and roridin A, was evaluated. In the first experiment, verrucarin A (1, 5, or 10 microg) and roridin A (5 or 10 microg) were each inoculated onto square-inch sections of glass, paper, and cloth and exposed to 1000 ppm of ClO2 for either 24 or 72 h at room temperature. In the second experiment, verrucarin A and roridin A (1 or 2 ppm in water) were treated with 200, 500, or 1000 ppm ClO2 for up to 116 h at room temperature in light and dark conditions (N = 9 per treatment for test and control). A yeast assay using Kluyveromyces marxianuswas used to quantify the toxicity of verrucarin A and roridin A. Additionally, high-performance liquid chromatography was performed on selected samples. Results for the first experiment showed that ClO2 treatment had no detectable effect on either toxin. For the second experiment, both toxins were completely inactivated at all tested concentrations in as little as 2 h after treatment with 1000 ppm ClO2. For verrucarin A, an effect was seen at the 500 ppm level, but this effect was not as strong as that observed at the 1000 ppm level. Roridin A toxicity was decreased after treatment with 200 and 500 ppm ClO2, but this was not significant until the 24-h exposure time was reached. These data show that ClO2 (in solution) can be effective for detoxification of roridin A or verrucarin A at selected concentrations and exposure times. PMID- 16040573 TI - The German Research Network on Depression and Suicidality: an introduction. PMID- 16040574 TI - The German Research Network on Depression and Suicidality. AB - Under the common roof of the German Research Network on Depression and Suicidality, scientists, medical doctors, psychotherapists, and several of the most relevant institutions of the German health care system are aligned to proceed against the overlapping health problems of depression and suicidality. The project addresses professionals and scientists, affected patients and their relatives, and the total population at the same time. Comprehensive public relations activities contribute to an overall sensitization concerning the topics of depression and suicidality and inform about diagnosis and treatment options. The network is funded by the German Ministry of Education and Research and was established in 1999. Since then more than 20 individual projects have been initiated. Meanwhile more and more results of these projects become available and increasing efforts are undertaken to transfer these results into elements of standard health care. PMID- 16040575 TI - How effective is advanced training concerning depression and suicidality among the elderly? Results of a pilot study. AB - Three Hundred and Seventy Four geriatric caregivers in Nuremberg and Munich participated in a four-hour advanced in-house training addressing the topic. "depression and suicidality in old age." Geriatric caregivers are key persons in early diagnosis of depressive symptoms and suicidality. The training focused on qualifying and sensitizing caregivers for these topics. A 12-item questionnaire was applied to measure the outcome parameters "knowledge and attitudes towards depression and suicidality in old age" within a pre-, post-, Three-months-follow up design (rate of return = 58%). Short-term effectiveness of the advanced training "depression in old age" could be shown for most of the items. Significant long-term effects could be found concerning increase of knowledge about pharmacotherapy with antidepressants. Attitude towards "suicide in old age" also changed significantly. The training program was effective in improving knowledge and attitudes of geriatric caregivers concerning depression and suicidality. PMID- 16040576 TI - A multicenter study about Neurobiology of Suicidal Behavior: design, development, and preliminary results. AB - The subproject 1.5 "Neurobiology of Suicidal Behavior" is a multicenter study assessing peripheral parameters of the serotonergic, noradrenergic, and dopaminergic transmitter systems. Additionally, stress hormones and the lipid system as well as inhibitory and excitatory amino acids will be investigated. The different parameters are collected in cerebral spinal fluid (CSF), blood, and saliva. Patients with a depressive spectrum disorder with and without a suicide attempt (during the last three weeks) and being medication free for two weeks are included in the study. So far, 103 patients and controls have been recruited. The design and development of this project as well as interconnections with the others subprojects are described. Preliminary results about the stress hormone system and suicidality are presented. PMID- 16040577 TI - Suicide prevention by lithium SUPLI--challenges of a multi-center prospective study. AB - Several studies have shown that there is a significantly increased risk of suicide related mortality in patients with a positive history of suicide attempts. The SUPLI-Study is the first prospective, randomized, double blind, placebo controlled multi-center trial focusing on the proposed suicide preventive effects of lithium in patients with suicidal behavior but not suffering from bipolar disorder or recurrent major depressive disorder. Patients with a recent history of a suicide attempt are treated with lithium versus placebo during a 12 month period. The hypothesis is that lithium treatment will lead to a 50% reduction of suicidal behavior. The protocol of the study and preliminary results are presented. PMID- 16040578 TI - Differential suicide rates in typologies of child sex offenders in a 6-year consecutive cohort of male suicides. AB - Earlier research identified 3 typologies of Child Sex Offenders [CSO] with high rates of suicide. To test this finding suicide rates of 3 types of CSO were compared in a 6-year cohort of regional suicides. All male suicides were identified from Coroners" inquest files and CSO data drawn from police records to calculate CSO suicide rates. The results show that suicide in "Multi-criminal" CSO is 12 times higher than the general population but not statistically significantly. Two significant results were "Sex Only" CSO suicides were 183 times the general population and 15 times the Multi-criminal CSO rates, with no suicides amongst the Violent CSO's. Implications for suicide prevention and child protection are presented. PMID- 16040579 TI - Suicidal ideation among help-seeking adolescents: association with a negative self-image. AB - Self-image and factors associated with suicidal ideation were studied among adolescents (N=168, age 17.7+/-2.3 years, 72% girls) seeking psychiatric assessment. The Structured Clinical Interview, Beck Depression Inventory and Offer Self-Image Questionnaire were used. Suicidal ideation (SI) was reported by 64% of subjects, of whom 20% received no psychiatric diagnosis and 58% had depressive disorders. SI in boys associated with being bullied at school and talking about one's problems only with friends, and in girls with a negative attitude towards the future and a negative self-image of one's own mental health. Because suicidal ideation frequently occurs without formal psychiatric diagnosis it should be assessed separately. Special attention should be paid to adolescents' negative self-perception. PMID- 16040580 TI - Race and method of suicide: culture and opportunity. AB - The present study assesses the link between choice of violent methods of suicide and race from the standpoint of two perspectives: differential socio acceptability and differential availability. To the extent that African Americans form a subculture of violence, and are more exposed to violence, we would expect them to choose violent methods of suicide. Data are from the 1990 mortality detail file of the U.S. Public Health Service and correspond to 19,580 male suicides. The results of a multivariate logistic regression analysis indicate that African Americans are 2.24 times more likely than Caucasians to choose violent methods of suicide. Although they are less likely to own firearms, African Americans are more likely than Caucasians to choose violent methods of suicide. PMID- 16040581 TI - Human immunodeficiency virus (HIV) and suicide in a region of Eastern Province ("Transkei"), South Africa. AB - HIV/AIDS is an alarming illness worldwide. It is known to have a significant association to suicide, despite a lack of in depth study. Early studies suggested suicide risk 20 to 36 times higher than the general population, but more recent trends in America show a decline. This is not true in Africa, including the region of Eastern Province, South Africa. There is abundant political and health discussion, but little study. This article presents descriptive statistics and case histories. These beginning studies must be augmented by comprehensive study of not only the epidemiology, but also the psychological and social correlates. Regrettably, there are numerous obstacles to such study in South Africa. PMID- 16040582 TI - Seasonal fluctuation of suicide in Hungary between 1970-2000. AB - The authors studied the seasonal fluctuation of suicide events in Hungary in a 31 year period. A summer maximum and a winter minimum were found in both genders. A second autumn peak did not occur among women. The suicide rate of Hungary fell markedly in 1998, parallel with the fundamental political-economical-social changes in the country. After 1988 the ANOVA statistic did show a moderate decrease in the seasonal fluctuation but only among those under 29 years of age. This young generation was the real "winner" of the deep social changes which suggests an indirect connection between the decrease of the seasonal fluctuation of suicide and the socio-political changes. PMID- 16040583 TI - Internet comments on media reporting of two adolescents' collective suicide attempt. AB - The presented study covered the analysis of 7 Internet media portrayals and 188 readers' spontaneous Internet comments about a case of two adolescents who survived a serious suicide attempt. Articles with photographs attracted the most responses (82%), irrespective of the length and content of the articles. More than half of the comments expressed readers' negative attitude (ironic 31% and angry 28%) while 21% of them remained neutral or indifferent. Twenty one percent of readers tried to initiate a serious discussion about the causes and prevention of suicide. An association was found between the style of media reporting and the number of readers' comments as well as certain aspects of the content of the comments. Inadequate reporting of suicidal behavior is associated with comments expressing oversimplified attitudes towards suicide. The influence of suicide reporting on the Internet on attitude formation in the general population should not be underestimated. PMID- 16040584 TI - Suicide and the Chinese Cultural Revolution. AB - This Commentary explores the impact of the Cultural Revolution instigated by Mao Zedong in Communist China from the year of 1966 to 1976 on suicidal behavior. Cases of individuals who completed suicide are presented, together with examples of the suicide notes they left. The lack of protest suicide is noted. PMID- 16040589 TI - Differences in ischemic lesion evolution in different rat strains using diffusion and perfusion imaging. AB - BACKGROUND AND PURPOSE: Interstrain differences in the temporal evolution of ischemia after middle cerebral artery occlusion (MCAO) in rats may considerably influence the results of experimental stroke research. We investigated, in 2 commonly used rat strains (Sprague-Dawley [SD] and Wistar-Kyoto [WK]), the spatiotemporal evolution of ischemia after permanent suture MCAO using diffusion and perfusion imaging. METHODS: Serial measurements of quantitative cerebral blood flow (CBF) and apparent diffusion coefficient (ADC) were performed up to 210 min after MCAO. Lesion volumes were calculated by using previously established viability thresholds and correlated with infarct volume defined by 2,3,5-triphenyltetrazolium chloride staining 24 hours after MCAO. RESULTS: While the ADC-derived lesion volume increased rapidly during the first 120 min after MCAO and essentially stopped growing after 3 hours in SD rats, ADC lesion in WK rats increased progressively during the entire 210-min period and was significantly smaller at all time points (P<0.05). The abnormal perfusion volume correlated highly with the TTC-defined infarct size in both groups. In WK rats, the abnormal perfusion volume was significantly larger than the abnormal diffusion volume up to 90 min after MCAO (P<0.001), whereas the diffusion/perfusion mismatch was significant (P<0.001) only at 45 min in SD rats. ADC-CBF scatterplots analysis revealed a slower and less robust ADC decline over time in WK rats in pixels with severe (<20% of normal) and moderate (21 to 40% of normal) CBF reduction. CONCLUSIONS: This study demonstrated substantial differences in acute ischemic lesion evolution between SD and WK rats. These interstrain variations must be taken into account when assessing new therapeutic approaches on ischemic lesion evolution in the rat MCAO model. PMID- 16040590 TI - Daily functioning and quality of life in a randomized controlled trial of therapeutic exercise for subacute stroke survivors. AB - BACKGROUND AND PURPOSE: The ability of therapeutic exercise after stroke to improve daily functioning and quality of life (QOL) remains controversial. We examined treatment effects on these outcomes in a randomized controlled trial (RCT) of exercise in subacute stroke survivors. METHODS: This is a secondary analysis of a single-blind RCT of a 12-week program versus usual care. Baseline, post-treatment and 6-month post-treatment daily functioning and QOL were assessed by Barthel index, Functional Independence Measure, instrumental activities of daily living, Medical Outcomes Study short-form 36-item questionnaire (SF-36), and Stroke Impact Scale (SIS). RESULTS: Of 100 randomized subjects, 93 completed the postintervention assessment, (mean age 70; 54% male; 81% white; mean Orpington Prognostic Score 3.4), and 80 had 6-month post-treatment assessment. Immediately after intervention, the intervention group improved more than usual care in SF-36 social function (14.0 points; P=0.0051) and in SIS (strength [9.2 points; P=0.0003], emotion [5.6 points; P=0.0240], social participation [6.6 points; P=0.0488], and physical function [5.0 points; P=0.0145]). Treatment was marginally more effective on Barthel score (3.3 points; P=0.0510), SF-36 (physical function [6.8 points; P=0.0586], physical role function [14.4 points; P=0.0708]), and SIS upper extremity function (7.2 points; P=0.0790). Effects were diluted 6 months after treatment ended. CONCLUSIONS: This rehabilitation exercise program led to more rapid improvement in aspects of physical, social, and role function than usual care in persons with subacute stroke. Adherence interventions to promote continued exercise after treatment might be needed to continue benefit. PMID- 16040591 TI - Evidence for age and sex differences in the secondary prevention of stroke in Scottish primary care. AB - BACKGROUND AND PURPOSE: Secondary preventive measures play an important role in the reduction of stroke, the third largest cause of death in Scotland. We investigated whether sex, age, or deprivation differences existed in the secondary prevention of stroke in primary care. METHODS: A retrospective cross sectional study using a computerized database with 61 practices (377,439 patients) to identify group differences in secondary preventive therapy between March 2003 and April 2004 for 10,076 patients with a diagnosis of any stroke. RESULTS: Women with any stroke were more likely than men to be prescribed a thiazide (odds ratios [OR], 1.60; 95% confidence interval [CI], 1.46 to 1.75) but less likely to be prescribed an angiotensin-converting enzyme inhibitor (OR, 0.73; 95% CI, 0.67 to 0.81). Women with ischemic stroke were less likely to receive either an antiplatelet or warfarin (OR, 0.84; 95% CI, 0.75 to 0.94) or statin therapy (OR, 0.82; 95% CI, 0.74 to 0.90) than men. Women with atrial fibrillation received less warfarin (OR, 0.62; 95% CI, 0.48 to 0.81) but more antiplatelet therapy than men (OR, 1.30; 95% CI, 1.00 to 1.68). The oldest patients (older than 75 years) with ischemic stroke received more antiplatelet therapy than the youngest patients (younger than 65 years) (OR, 1.83; 95% CI, 1.64 to 2.06). No significant differences in secondary preventative treatment across deprivation groups were found. CONCLUSIONS: Important sex and age differences exist in the care of patients with stroke and suggest that women and the elderly need to be targeted for secondary prevention therapy. PMID- 16040592 TI - Neonatal hypoxia/ischemia is associated with decreased inflammatory mediators after erythropoietin administration. AB - BACKGROUND AND PURPOSE: Erythropoietin (EPO), a hematopoietic growth factor, has been shown to be neuroprotective when administered as either a pretreatment or posttreatment. This study tested the hypothesis that one of the mechanisms of protection afforded by posttreatment with recombinant human EPO (rh-EPO) is an anti-inflammatory effect via inhibition of interleukin (IL)-1beta. METHODS: Seven day-old rat pups were subjected to unilateral carotid artery ligation followed by 90 minutes of hypoxia (8% O2 at 37 degrees C). Pups were divided into the following groups: control, hypoxia/ischemia, and hypoxia/ischemia plus rh-EPO. In the rh-EPO-treated pups, rh-EPO (5 U/g body weight IP) was administered starting 24 hours after the insult and then for 2 additional days. Samples were collected at 3, 7, 14, and 21 days after the insult. IL-1beta mRNA and protein levels were determined by quantitative real-time reverse transcription-polymerase chain reaction and ELISA. Tumor necrosis factor (TNF)-alpha mRNA levels were determined by colorimetric microplate assay. RESULTS: rhEPO attenuated brain injury, as assessed by brain weight, and attenuated both the hypoxia/ischemia-induced increases in IL-1beta mRNA and protein levels. TNF-alpha mRNA levels did not increase at 3 to 14 days after the hypoxic/ischemic insult. CONCLUSIONS: Administration of exogenous rh-EPO starting 24 hours after a hypoxic/ischemic insult is neuroprotective in the neonatal rat. This neuroprotective activity prevented the secondary, delayed rise in IL-1beta and attenuated the infiltration of leukocytes into the ipsilateral hemisphere. PMID- 16040593 TI - Interaction between a rat model of cerebral ischemia and beta-amyloid toxicity: II. Effects of triflusal. AB - BACKGROUND AND PURPOSE: Clinical data suggest that Alzheimer disease (AD) and stroke together potentiate cognitive impairment. Our rat model demonstrates that this interaction may be mediated through inflammatory cells and pathways. Thus, anti-inflammatory agents such as Triflusal, a nonsteroidal anti-inflammatory agent (NSAID), may provide neuroprotection for susceptible neurons in AD and cerebral ischemia. METHODS: AD was modeled by cerebroventricular injections of beta-amyloid (Abeta25-35) and subcortical lacunar infarcts by striatal endothelin injections. Inflammatory mechanisms were examined by immunohistochemical analysis. Behavioral tasks were assessed with the Montoya staircase test. RESULTS: Triflusal reduced pathologic and inflammatory markers and functional deficits in rats receiving Abeta or endothelin alone but was less effective in the more severe pathology of the combined Abeta/endothelin model. CONCLUSIONS: Higher doses or more prolonged treatment with NSAIDs may be required for more effective neuroprotection in combined AD and stroke conditions. PMID- 16040594 TI - Ambulatory transcranial Doppler cerebral embolic signal detection in symptomatic and asymptomatic carotid stenosis. AB - BACKGROUND AND PURPOSE: Transcranial Doppler (TCD) ultrasound can detect asymptomatic emboli in carotid stenosis. Current systems are nonportable and can only record for short durations. A novel ambulatory TCD system allows prolonged recording. We applied this to patients with symptomatic and asymptomatic carotid stenosis to determine patterns of embolization in the 2 conditions and optimal recording protocols. METHODS: Ambulatory TCD recordings were performed in 12 symptomatic and 15 asymptomatic carotid stenosis (> or =50%) patients for 8 hours and then repeated on a second occasion. RESULTS: Nine (75%) of symptomatic subjects had embolic signals during the first recording. In this group, repeating the recording did not increase the proportion of positive patients. In asymptomatic patients, 4 (26.7%) had embolic signals on 1 recording, and this proportion increased to 46.7% after 2 recordings. There was significant clustering of embolic signals demonstrating that the process was nonrandom. CONCLUSIONS: Ambulatory TCD is possible in patients with carotid artery stenosis. By increasing the duration of recording, additional information is provided, particularly in asymptomatic patients. Our results also demonstrate clustering of embolic signals. Our study provides baseline data to allow studies in both asymptomatic and symptomatic carotid stenosis to be planned. PMID- 16040595 TI - Discovery of a novel restriction endonuclease by genome comparison and application of a wheat-germ-based cell-free translation assay: PabI (5'-GTA/C) from the hyperthermophilic archaeon Pyrococcus abyssi. AB - To search for restriction endonucleases, we used a novel plant-based cell-free translation procedure that bypasses the toxicity of these enzymes. To identify candidate genes, the related genomes of the hyperthermophilic archaea Pyrococcus abyssi and Pyrococcus horikoshii were compared. In line with the selfish mobile gene hypothesis for restriction-modification systems, apparent genome rearrangement around putative restriction genes served as a selecting criterion. Several candidate restriction genes were identified and then amplified in such a way that they were removed from their own translation signal. During their cloning into a plasmid, the genes became connected with a plant translation signal. After in vitro transcription by T7 RNA polymerase, the mRNAs were separated from the template DNA and translated in a wheat-germ-based cell-free protein synthesis system. The resulting solution could be directly assayed for restriction activity. We identified two deoxyribonucleases. The novel enzyme was denoted as PabI, purified and found to recognize 5'-GTAC and leave a 3'-TA overhang (5'-GTA/C), a novel restriction enzyme-generated terminus. PabI is active up to 90 degrees C and optimally active at a pH of around 6 and in NaCl concentrations ranging from 100 to 200 mM. We predict that it has a novel 3D structure. PMID- 16040596 TI - Four new type I restriction enzymes identified in Escherichia coli clinical isolates. AB - Using a plasmid transformation method and the RM search computer program, four type I restriction enzymes with new recognition sites and two isoschizomers (EcoBI and Eco377I) were identified in a collection of clinical Escherichia coli isolates. These new enzymes were designated Eco394I, Eco826I, Eco851I and Eco912I. Their recognition sequences were determined to be GAC(5N)RTAAY, GCA(6N)CTGA, GTCA(6N)TGAY and CAC(5N)TGGC, respectively. A methylation sensitivity assay, using various synthetic oligonucleotides, was used to identify the adenines that prevent cleavage when methylated (underlined). These results suggest that type I enzymes are abundant in E.coli and many other bacteria, as has been inferred from bacterial genome sequencing projects. PMID- 16040597 TI - Discovery of the principal specific transcription factors of Apicomplexa and their implication for the evolution of the AP2-integrase DNA binding domains. AB - The comparative genomics of apicomplexans, such as the malarial parasite Plasmodium, the cattle parasite Theileria and the emerging human parasite Cryptosporidium, have suggested an unexpected paucity of specific transcription factors (TFs) with DNA binding domains that are closely related to those found in the major families of TFs from other eukaryotes. This apparent lack of specific TFs is paradoxical, given that the apicomplexans show a complex developmental cycle in one or more hosts and a reproducible pattern of differential gene expression in course of this cycle. Using sensitive sequence profile searches, we show that the apicomplexans possess a lineage-specific expansion of a novel family of proteins with a version of the AP2 (Apetala2)-integrase DNA binding domain, which is present in numerous plant TFs. About 20-27 members of this apicomplexan AP2 (ApiAP2) family are encoded in different apicomplexan genomes, with each protein containing one to four copies of the AP2 DNA binding domain. Using gene expression data from Plasmodium falciparum, we show that guilds of ApiAP2 genes are expressed in different stages of intraerythrocytic development. By analogy to the plant AP2 proteins and based on the expression patterns, we predict that the ApiAP2 proteins are likely to function as previously unknown specific TFs in the apicomplexans and regulate the progression of their developmental cycle. In addition to the ApiAP2 family, we also identified two other novel families of AP2 DNA binding domains in bacteria and transposons. Using structure similarity searches, we also identified divergent versions of the AP2-integrase DNA binding domain fold in the DNA binding region of the PI-SceI homing endonuclease and the C-terminal domain of the pleckstrin homology (PH) domain-like modules of eukaryotes. Integrating these findings, we present a reconstruction of the evolutionary scenario of the AP2-integrase DNA binding domain fold, which suggests that it underwent multiple independent combinations with different types of mobile endonucleases or recombinases. It appears that the eukaryotic versions have emerged from versions of the domain associated with mobile elements, followed by independent lineage-specific expansions, which accompanied their recruitment to transcription regulation functions. PMID- 16040598 TI - Length-dependent energetics of (CTG)n and (CAG)n trinucleotide repeats. AB - Trinucleotide repeats are involved in a number of debilitating diseases such as myotonic dystrophy. Twelve to seventy-five base-long (CTG)n oligodeoxynucleotides were analysed using a combination of biophysical [UV-absorbance, circular dichroism and differential scanning calorimetry (DSC)] and biochemical methods (non-denaturing gel electrophoresis and enzymatic footprinting). All oligomers formed stable intramolecular structures under near physiological conditions with a melting temperature that was only weakly dependent on oligomer length. Thermodynamic analysis of the denaturation process by UV-melting and calorimetric experiments revealed an unprecedented length-dependent discrepancy between the enthalpy values deduced from model-dependent (UV-melting) and model-independent (calorimetry) experiments. Evidence for non-zero molar heat capacity changes was also derived from the analysis of the Arrhenius plots and DSC profiles. Such behaviour is analysed in the framework of an intramolecular 'branched-hairpin' model, in which long CTG oligomers do not fold into a simple long hairpin-stem intramolecular structure, but allow the formation of several independent folding units of unequal stability. We demonstrate that, for sequences ranging from 12 to 25 CTG repeats, an intramolecular structure with two loops is formed which we will call 'bis-hairpin'. Similar results were also found for CAG oligomers, suggesting that this observation may be extended to various trinucleotide repeats containing sequences. PMID- 16040599 TI - Contrasting effects of Elg1-RFC and Ctf18-RFC inactivation in the absence of fully functional RFC in fission yeast. AB - Proliferating cell nuclear antigen loading onto DNA by replication factor C (RFC) is a key step in eukaryotic DNA replication and repair processes. In this study, the C-terminal domain (CTD) of the large subunit of fission yeast RFC is shown to be essential for its function in vivo. Cells carrying a temperature-sensitive mutation in the CTD, rfc1-44, arrest with incompletely replicated chromosomes, are sensitive to DNA damaging agents, are synthetically lethal with other DNA replication mutants, and can be suppressed by mutations in rfc5. To assess the contribution of the RFC-like complexes Elg1-RFC and Ctf18-RFC to the viability of rfc1-44, genes encoding the large subunits of these complexes have been deleted and overexpressed. Inactivation of Ctf18-RFC by the deletion of ctf18+, dcc1+ or ctf8+ is lethal in an rfc1-44 background showing that full Ctf18-RFC function is required in the absence of fully functional RFC. In contrast, rfc1-44 elg1Delta cells are viable and overproduction of Elg1 in rfc1-44 is lethal, suggesting that Elg1-RFC plays a negative role when RFC function is inhibited. Consistent with this, the deletion of elg1+ is shown to restore viability to rfc1-44 ctf18Delta cells. PMID- 16040600 TI - Deletion of Cg-emb in corynebacterianeae leads to a novel truncated cell wall arabinogalactan, whereas inactivation of Cg-ubiA results in an arabinan-deficient mutant with a cell wall galactan core. AB - The cell wall of Mycobacterium tuberculosis has a complex ultrastructure that consists of mycolic acids connected to peptidoglycan via arabinogalactan (AG) and abbreviated as the mAGP complex. The mAGP complex is crucial for the survival and pathogenicity of M. tuberculosis and is the target of several anti-tubercular agents. Apart from sharing a similar mAGP and the availability of the complete genome sequence, Corynebacterium glutamicum has proven useful in the study of orthologous M. tuberculosis genes essential for viability. Here we examined the effects of particular genes involved in AG polymerization by gene deletion in C. glutamicum. The anti-tuberculosis drug ethambutol is thought to target a set of arabinofuranosyltransferases (Emb) that are involved in arabinan polymerization. Deletion of emb in C. glutamicum results in a slow growing mutant with profound morphological changes. Chemical analysis revealed a dramatic reduction of arabinose resulting in a novel truncated AG structure possessing only terminal arabinofuranoside (t-Araf) residues with a corresponding loss of cell wall bound mycolic acids. Treatment of wild-type C. glutamicum with ethambutol and subsequent cell wall analyses resulted in an identical phenotype comparable to the C. glutamicum emb deletion mutant. Additionally, disruption of ubiA in C. glutamicum, the first enzyme involved in the biosynthesis of the sugar donor decaprenol phosphoarabinose (DPA), resulted in a complete loss of cell wall arabinan. Herein, we establish for the first time, (i) that in contrast to M. tuberculosis embA and embB mutants, deletion of C. glutamicum emb leads to a highly truncated AG possessing t-Araf residues, (ii) the exact site of attachment of arabinan chains in AG, and (iii) DPA is the only Araf sugar donor in AG biosynthesis suggesting the presence of a novel enzyme responsible for "priming" the galactan domain for further elaboration by Emb, resulting in the final maturation of the native AG polysaccharide. PMID- 16040601 TI - Defining the role of ubiquitin-interacting motifs in the polyglutamine disease protein, ataxin-3. AB - Polyglutamine (polyQ) expansions cause neurodegeneration that is associated with protein misfolding and influenced by functional properties of the host protein. The polyQ disease protein, ataxin-3, has predicted ubiquitin-specific protease and ubiquitin-binding domains, which suggest that ataxin-3 functions in ubiquitin dependent protein surveillance. Here we investigate direct links between the ubiquitin-proteasome pathway and ataxin-3. In neural cells we show that, through its ubiquitin interaction motifs (UIMs), normal or expanded ataxin-3 binds a broad range of ubiquitinated proteins that accumulate when the proteasome is inhibited. The expression of a catalytically inactive ataxin-3 (normal or expanded) causes ubiquitinated proteins to accumulate in cells, even in the absence of proteasome inhibitor. This accumulation of ubiquitinated proteins occurs primarily in the cell nucleus in transfected cells and requires intact UIMs in ataxin-3. We further show that both normal and expanded ataxin-3 can undergo oligoubiquitination. Although this post-translational modification occurs in a UIM-dependent manner, it becomes independent of UIMs when the catalytic cysteine residue of ataxin-3 is mutated, suggesting that ataxin-3 ubiquitination is itself regulated in trans by its own de-ubiquitinating activity. Finally, pulse-chase labeling reveals that ataxin-3 is degraded by the proteasome, with expanded ataxin-3 being as efficiently degraded as normal ataxin-3. Mutating the UIMs does not alter degradation, suggesting that UIM-mediated oligoubiquitination of ataxin-3 modulates ataxin-3 function rather than stability. The function of ataxin-3 as a de-ubiquitinating enzyme, its post-translational modification by ubiquitin, and its degradation via the proteasome link this polyQ protein to ubiquitin-dependent pathways already implicated in disease pathogenesis. PMID- 16040602 TI - A true autoactivating enzyme. Structural insight into mannose-binding lectin associated serine protease-2 activations. AB - Few reports have described in detail a true autoactivation process, where no extrinsic cleavage factors are required to initiate the autoactivation of a zymogen. Herein, we provide structural and mechanistic insight into the autoactivation of a multidomain serine protease: mannose-binding lectin associated serine protease-2 (MASP-2), the first enzymatic component in the lectin pathway of complement activation. We characterized the proenzyme form of a MASP-2 catalytic fragment encompassing its C-terminal three domains and solved its crystal structure at 2.4 A resolution. Surprisingly, zymogen MASP-2 is capable of cleaving its natural substrate C4, with an efficiency about 10% that of active MASP-2. Comparison of the zymogen and active structures of MASP-2 reveals that, in addition to the activation domain, other loops of the serine protease domain undergo significant conformational changes. This additional flexibility could play a key role in the transition of zymogen MASP-2 into a proteolytically active form. Based on the three-dimensional structures of proenzyme and active MASP-2 catalytic fragments, we present model for the active zymogen MASP-2 complex and propose a mechanism for the autoactivation process. PMID- 16040603 TI - Streptococcus pyogenes collagen type I-binding Cpa surface protein. Expression profile, binding characteristics, biological functions, and potential clinical impact. AB - The Streptococcus pyogenes collagen type I-binding protein Cpa (collagen-binding protein of group A streptococci) expressed by 28 serotypes of group A streptococci has been extensively characterized at the gene and protein levels. Evidence for three distinct families of cpa genes was found, all of which shared a common sequence encoding a 60-amino acid domain that accounted for selective binding to type I collagen. Surface plasmon resonance-based affinity measurements and functional studies indicated that the expression of Cpa was consistent with an attachment role for bacteria to tissue containing collagen type I. A cpa mutant displayed a significantly decreased internalization rate when incubated with HEp-2 cells but had no effect on the host cell viability. By utilizing serum from patients with a positive titer for streptolysin/DNase antibody, an increased anti-Cpa antibody titer was noted for patients with a clinical history of arthritis or osteomyelitis. Taken together, these results suggest Cpa may be a relevant matrix adhesin contributing to the pathogenesis of S. pyogenes infection of bones and joints. PMID- 16040604 TI - The Lactococcus lactis CodY regulon: identification of a conserved cis-regulatory element. AB - CodY of Lactococcus lactis MG1363 is a transcriptional regulator that represses the expression of several genes encoding proteins of the proteolytic system. DNA microarray analysis, comparing the expression profiles of L. lactis MG1363 and an isogenic strain in which codY was mutated, was used to determine the CodY regulon. In peptide-rich medium and exponentially growing cells, where CodY exerts strong repressing activity, the expression of over 30 genes was significantly increased upon removal of codY. The differentially expressed genes included those predominantly involved in amino acid transport and metabolism. In addition, several genes belonging to other functional categories were derepressed, stressing the pleiotropic role of CodY. Scrutinizing the transcriptome data with bioinformatics tools revealed the presence of a novel over-represented motif in the upstream regions of several of the genes derepressed in L. lactis MG1363DeltacodY. Evidence is presented that this 15-bp cis-sequence, AATTTTCWGAAAATT, serves as a high affinity binding site for CodY, as shown by electrophoretic mobility shift assays and DNase I footprinting analyses. The presence of this CodY-box is sufficient to evoke CodY-mediated regulation in vivo. A copy of this motif is also present in the upstream region of codY itself. It is shown that CodY regulates its own synthesis and requires the CodY-box and branched-chain amino acids to interact with its promoter. PMID- 16040605 TI - Group V secretory phospholipase A2-modified low density lipoprotein promotes foam cell formation by a SR-A- and CD36-independent process that involves cellular proteoglycans. AB - Accumulating evidence indicates that secretory phospholipase A2 (sPLA2) enzymes promote atherogenic processes. We have previously showed the presence of Group V sPLA2 (GV sPLA2) in human and mouse atherosclerotic lesions, its hydrolysis of low density lipoprotein (LDL) particles, and the ability of GV sPLA2-modified LDL (GV-LDL) to induce macrophage foam cell formation in vitro. The goal of this study was to investigate the mechanisms involved in macrophage uptake of GV-LDL. Peritoneal macrophages from C57BL/6 mice (wild type (WT)), C57BL/6 mice deficient in LDL receptor (LDLR-/-), or SR-A and CD36 (DKO) were treated with control LDL, GV-LDL, oxidized LDL (ox-LDL) or LDL aggregated by vortexing (vx-LDL). As expected, ox-LDL induced significantly more cholesterol ester accumulation in WT and LDLR-/- compared with DKO macrophages. In contrast, there was no difference in the accumulation of GV-LDL or vx-LDL in the three cell types. 125I-ox-LDL exhibited high affinity, saturable binding to WT cells that was significantly reduced in DKO cells. Vx-LDL and GV-LDL showed low affinity, non-saturable binding that was similar for both cell types, and significantly higher compared with control LDL. GV-LDL degradation in WT and DKO cells was similar. Analyses by confocal microscopy indicated a distinct intracellular distribution of Alexa-568 labeled GV-LDL and Alexa-488-labeled ox-LDL. Uptake of GV-LDL (but not ox-LDL or vx-LDL) was significantly reduced in cells preincubated with heparin or NaClO3, suggesting a role for proteoglycans in GV-LDL uptake. Our data point to a physiological modification of LDL that has the potential to promote macrophage foam cell formation independent of scavenger receptors. PMID- 16040606 TI - Possible role of direct Rac1-Rab7 interaction in ruffled border formation of osteoclasts. AB - Rab7 has been shown to regulate the late steps of the endocytic pathway. In bone resorbing osteoclasts, it is involved in formation of the ruffled border, which is a late endosomal-like compartment in the plasma membrane. Here we report a new Rab7-interacting protein, Rac1, another small GTPase protein that binds to the GTP-form of Rab7 as found with a two-hybrid system. We demonstrate further that Rab7 colocalizes with Rac1 at the fusion zone of the ruffled border in bone resorbing osteoclasts. In other cell types, such as fibroblast-like cells, partial colocalization is perinuclear. Because Rac1 is known to control the actin cytoskeleton through its effectors, the Rab7-Rac1 interaction may mediate late endosomal transport between microtubules and microfilaments enabling endosomal vesicles to switch tracks and may thus also regulate ruffled border formation in osteoclasts. PMID- 16040607 TI - Identification of a gene involved in polysaccharide export as a transcription target of FruA, an essential factor for Myxococcus xanthus development. AB - Fruiting body development in Myxococcus xanthus is a multicellular event that is coordinated by exchanging intercellular signals. FruA is a transcription factor essential for fruiting body development and is thought to play a key role in the C-signal pathway. Here we present the first identification of a gene regulated by FruA. The gene was isolated from a genomic library via in vitro selection in a DNA binding assay by using the DNA-binding domain of FruA tagged with His(8) at the C-terminal end (FruA-DBD-H(8)). The gene, named fdgA (FruA-dependent gene A), encodes a protein homologous to the outer-membrane auxiliary family protein involved in the polysaccharide export system. FruA-DBD-H(8) bound the upstream promoter region of the fdgA gene from nucleotide -89 to nucleotide -64 with respect to the transcription initiation site, which was required for the induction of fdgA expression during development. fdgA mRNA induced during development was absent in a fruA deletion strain. The deletion of fdgA resulted in defective fruiting body formation and reduced sporulation efficiency (1% that of the parent strain). Moreover, FruA was required for the developmental expression of sasA, which is also involved in the biosynthesis of the lipopolysaccharide O-antigen and is required for fruiting body development. Furthermore, the expression of both fdgA and sasA was partially dependent on the C-signal. These findings expand our understanding of the signal transduction pathway mediated by FruA during development in M. xanthus. PMID- 16040608 TI - Rapid effects of dexamethasone on intracellular pH and Na+/H+ exchanger activity in human bronchial epithelial cells. AB - Glucocorticoids have been shown to produce rapid nongenomic responses in airway epithelia. By using an intracellular pH (pH(i)) spectrofluorescence imaging system and the NH4Cl acid-loading technique, we have shown that the synthetic glucocorticoid,dexamethasone, accelerated intracellular pH recovery after an acid load in a human bronchial epithelial cell line (16HBE14o- cells). Exposure to NH4Cl (20 mm) elicited an intracellular acidification, followed by a pH(i) recovery. Inhibition of the Na+/H+ exchanger decreased the steady-state pH(i) and antagonized the dexamethasone stimulation of pH(i) regulation. The rapid effect of dexamethasone on pH(i) was neither affected by the inhibitor of transcription, cycloheximide, nor by the classical glucocorticoid and mineralocorticoid receptors antagonists, RU486 and spironolactone, respectively. The dexamethasone effect on pH(i) regulation was reduced by inhibitors of adenylate cyclase, cAMP dependent protein kinase and mitogenactivated protein kinase (ERK1/2). By using a PepTag assay system and Western blotting, we have shown that dexamethasone stimulated cAMP-dependent protein kinase and mitogen-activated protein kinase activities. Taken together our results provide evidence for the rapid stimulation of Na+/H+ exchange activity by glucocorticoids in bronchial epithelial cells via a nongenomic mechanism involving cAMP-dependent protein kinase and mitogen activated protein kinase ERK1/2 pathways. PMID- 16040609 TI - STN8 protein kinase in Arabidopsis thaliana is specific in phosphorylation of photosystem II core proteins. AB - Combination of reversed genetics with analyses of in vivo protein phosphorylation in Arabidopsis thaliana revealed that STN8 protein kinase is specific in phosphorylation of N-terminal threonine residues in D1, D2, and CP43 proteins, and Thr-4 in the PsbH protein of photosystem II. Phosphorylation of D1, D2, and CP43 in the light-exposed leaves of two Arabidopsis lines with T-DNA insertions in the stn8 gene was found significantly reduced in the assays with anti phosphothreonine antibodies. Protein phosphorylation in each of the mutants was quantified comparatively to the wild type by mass spectrometric analyses of phosphopeptides released from the photosynthetic membranes and differentially labeled with stable isotopes. The lack of STN8 caused 50-60% reduction in D1 and D2 phosphorylation, but did not change the phosphorylation level of two peptides that could correspond to light-harvesting proteins encoded by seven different genes in Arabidopsis. Phosphorylation of the PsbH protein at Thr-4 was completely abolished in the plants lacking STN8. Phosphorylation of Thr-4 in the wild type required both light and prior phosphorylation at Thr-2, indicating that STN8 is a light-activated kinase that phosphorylates Thr-4 only after another kinase phosphorylates Thr-2. Analysis of the STN8 catalytic domain suggests that selectivity of STN8 in phosphorylation of the very N-terminal residues in D1, D2, and CP43, and Thr-4 in PsbH pre-phosphorylated at Thr-2 may be explained by the long loops obstructing entrance into the kinase active site and seven additional basic residues in the vicinity of the catalytic site, as compared with the homologous STN7 kinase responsible for phosphorylation of light-harvesting proteins. PMID- 16040610 TI - Anillin is a substrate of anaphase-promoting complex/cyclosome (APC/C) that controls spatial contractility of myosin during late cytokinesis. AB - Anillin, an actin-binding protein localized at the cleavage furrow, is required for cytokinesis. Through an in vitro expression screen, we identified anillin as a substrate of the anaphase-promoting complex/cyclosome (APC/C), a ubiquitin ligase that controls mitotic progression. We found that the levels of anillin fluctuate in the cell cycle, peaking in mitosis and dropping drastically during mitotic exit. Ubiquitination of anillin required a destruction-box and was mediated by Cdh1, an activator of APC/C. Overexpression of Cdh1 reduced the levels of anillin, whereas inactivation of APC/C(Cdh1) increased the half-life of anillin. Functionally, anillin was required for the completion of cytokinesis. In anillin knockdown cells, the cleavage furrow ingressed but failed to complete the ingression. At late cytokinesis, the cytosol and DNA in knockdown cells underwent rapid myosin-based oscillatory movement across the furrow. During this movement, RhoA and active myosin were absent from the cleavage furrow, and myosin was redistributed to cortical patches, which powers the random oscillatory movement. We concluded that anillin functions to maintain the localization of active myosin, thereby ensuring the spatial control of concerted contraction during cytokinesis. PMID- 16040611 TI - A bacterial glutathione transporter (Escherichia coli CydDC) exports reductant to the periplasm. AB - Glutathione (GSH), a major biological antioxidant, maintains redox balance in prokaryotes and eukaryotic cells and forms exportable conjugates with compounds of pharmacological and agronomic importance. However, no GSH transporter has been characterized in a prokaryote. We show here that a heterodimeric ATP-binding cassette-type transporter, CydDC, mediates GSH transport across the Escherichia coli cytoplasmic membrane. In everted membrane vesicles, GSH is imported via an ATP-driven, protonophore-insensitive, orthovanadate-sensitive mechanism, equating with export to the periplasm in intact cells. GSH transport and cytochrome bd quinol oxidase assembly are abolished in the cydD1 mutant. Glutathione disulfide (GSSG) was not transported in either Cyd(+) or Cyd(-) strains. Exogenous GSH restores defective swarming motility and benzylpenicillin sensitivity in a cydD mutant and also benzylpenicillin sensitivity in a gshA mutant defective in GSH synthesis. Overexpression of the cydDC operon in dsbD mutants defective in disulfide bond formation restores dithiothreitol tolerance and periplasmic cytochrome b assembly, revealing redundant pathways for reductant export to the periplasm. These results identify the first prokaryotic GSH transporter and indicate a key role for GSH in periplasmic redox homeostasis. PMID- 16040612 TI - Redox-induced protein structural changes in cytochrome bo revealed by Fourier transform infrared spectroscopy and [13C]Tyr labeling. AB - Cytochrome bo is a heme-copper terminal ubiquinol oxidase of Escherichia coli under highly aerated growth conditions. Tyr-288 present at the end of the K channel forms a Cepsilon-Nepsilon covalent bond with one of the Cu(B) ligand histidines and has been proposed to be an acid-base catalyst essential for the O O bond cleavage at the Oxy-to-P transition of the dioxygen reduction cycle (Uchida, T., Mogi, T., and Kitagawa, T. (2000) Biochemistry 39, 6669-6678). To probe structural changes at tyrosine residues, we examined redox difference Fourier transform infrared difference spectra of the wild-type enzyme in which either L-[1-13C]Tyr or L-[4-13C]Tyr has been biosynthetically incorporated in the tyrosine auxotroph. Spectral comparison between [1-13C]Tyr-labeled and unlabeled proteins indicated that substitution of the main chain carbonyl of a Tyr residue(s) significantly affected changes in the amide-I (approximately 1620-1680 cm(-1)) and -II ( approximately 1540-1560 cm(-1)) regions. In contrast, spectral comparison between [4-13C]Tyr-labeled and unlabeled proteins showed only negligible changes, which was the case for both the pulsed and the resting forms. Thus, protonation of an OH group of tyrosines including Tyr-288 in the vicinity of the heme o-Cu(B) binuclear center was not detected at pH 7.4 upon full reduction of cytochrome bo. Redox-induced main chain changes at a Tyr residue(s) are associated with structural changes at Glu-286 near the binuclear metal centers and may be related to switching of the K-channel operative at the reductive phase to D-channel at the oxidative phase of the dioxygen reduction cycle via conformational changes in the middle of helix VI. PMID- 16040613 TI - Feedback regulation of murine pantothenate kinase 3 by coenzyme A and coenzyme A thioesters. AB - Pantothenate kinase catalyzes a key regulatory step in coenzyme A biosynthesis, and there are four mammalian genes that encode isoforms of this enzyme. Pantothenate kinase isoform PanK3 is highly related to the previously characterized PanK1beta isoform (79% identical, 91% similar), and these two almost identical proteins are expressed most highly in the same tissues. PanK1beta and PanK3 had very similar molecular sizes, oligomeric form, cytoplasmic cellular location, and kinetic constants for ATP and pantothenate. However, these two PanK isoforms possessed distinct regulatory properties. PanK3 was significantly more sensitive to feedback regulation by acetyl-CoA (IC50 = 1 microm) than PanK1beta (IC50 = 10 microm), and PanK3 was stringently regulated by long-chain acyl-CoA (IC50 = 2 microm), whereas PanK1beta was not. Domain swapping experiments localized the difference in the two proteins to a 48-amino-acid domain, where they are the most divergent. Consistent with these more stringent regulatory properties, metabolic labeling experiments showed that coenzyme A (CoA) levels in cells overexpressing PanK3 were lower than in cells overexpressing an equivalent amount of PanK1beta. Thus, the distinct regulatory properties exhibited by the family of the pantothenate kinases allowed the rate of CoA biosynthesis to be controlled by regulatory signals from CoA thioesters involved in different branches of intermediary metabolism. PMID- 16040614 TI - Probing the mechanism of the Mycobacterium tuberculosis beta-ketoacyl-acyl carrier protein synthase III mtFabH: factors influencing catalysis and substrate specificity. AB - Mycolic acids are the dominant feature of the Mycobacterium tuberculosis cell wall. These alpha-alkyl, beta-hydroxy fatty acids are formed by the condensation of two fatty acids, a long meromycolic acid and a shorter C(24)-C(26) fatty acid. The component fatty acids are produced via a combination of type I and II fatty acid synthases (FAS) with FAS-I products being elongated by FAS-II toward meromycolic acids. The beta-ketoacyl-acyl carrier protein (ACP) synthase III encoded by mtfabH (mtFabH) links FAS-I and FAS-II, catalyzing the condensation of FAS-I-derived acyl-CoAs with malonyl-acyl carrier protein (ACP). The acyl-CoA chain length specificity of mtFabH was assessed in vitro; the enzyme extended longer, physiologically relevant acyl-CoA primers when paired with AcpM, its natural partner, than with Escherichia coli ACP. The ability of the enzyme to use E. coli ACP suggests that a similar mode of binding is likely with both ACPs, yet it is clear that unique factors inherent to AcpM modulate the substrate specificity of mtFabH. Mutation of proposed key mtFabH residues was used to define their catalytic roles. Substitution of supposed acyl-CoA binding residues reduced transacylation, with double substitutions totally abrogating activity. Mutation of Arg(46) revealed its more critical role in malonyl-AcpM decarboxylation than in the acyl-CoA binding role. Interestingly, this effect was suppressed intragenically by Arg(161) --> Ala substitution. Our structural studies suggested that His(258), previously implicated in malonyl-ACP decarboxylation, also acts as an anchor point for a network of water molecules that we propose promotes deprotonation and transacylation of Cys(122). PMID- 16040616 TI - High mobility group protein-B1 interacts with sterol regulatory element-binding proteins to enhance their DNA binding. AB - Sterol regulatory element-binding proteins (SREBPs) are transcription factors that are predominately involved in the regulation of lipogenic and cholesterogenic enzyme gene expression. To identify unknown proteins that interact with SREBP, we screened nuclear extract proteins with 35S-labeled SREBP 1 bait in Far Western blotting analysis. Using this approach, high mobility group protein-B1 (HMGB1), a chromosomal protein, was identified as a novel SREBP interacting protein. In vitro glutathione S-transferase pull-down and in vivo coimmunoprecipitation studies confirmed an interaction between HMGB1 and both SREBP-1 and -2. The protein-protein interaction was mediated through the helix loop-helix domain of SREBP-1, residues 309-344, and the A box of HMGB1. Furthermore, an electrophoretic mobility shift assay demonstrated that HMGB1 enhances SREBPs binding to their cognate DNA sequences. Moreover, luciferase reporter analyses, including RNA interference technique showed that HMGB1 potentiates the transcriptional activities of SREBP in cultured cells. These findings raise the intriguing possibility that HMGB1 is potentially involved in the regulation of lipogenic and cholesterogenic gene transcription. PMID- 16040618 TI - What is the normal range for N-terminal pro-brain natriuretic peptide? How well does this normal range screen for cardiovascular disease? AB - AIMS: To define the N-terminal pro-brain natriuretic peptide (NTpBNP) normal range, assessing its cardiovascular screening characteristics in general population and higher risk subjects. METHODS AND RESULTS: A total of 2320 subjects (1392 general population and 928 high-risk) > or =45 years old, selected randomly from seven community practices, were invited to undergo clinical assessment and echocardiography and to assess NTpBNP serum levels. Of these, 1205 attended. The NTpBNP normal range was calculated and its cardiovascular screening characteristics were assessed. Age (P<0.0001) and female gender (P<0.0001) independently predicted NTpBNP levels in normal subjects. In the general population, age- and gender-stratified normal NTpBNP levels gave a negative predictive value (NPV) of 99% in excluding left ventricular systolic dysfunction, atrial fibrillation, and valvular heart disease, and a positive predictive value of 56% in detecting any cardiovascular disease assessed. In high-risk subjects, these values were 98 and 62%, respectively. Ninety-five per cent of subjects with NTpBNP levels over four times the normal had significant cardiovascular disease with the others having renal dysfunction. CONCLUSION: Normal NTpBNP levels should be stratified by age and gender. Normal NTpBNP levels give high NPV in excluding significant cardiovascular disease. Most subjects with raised NTpBNP levels and almost all subjects with NTpBNP levels over four times the normal have significant cardiovascular disease. PMID- 16040619 TI - Influence of heart rate, blood pressure, and beta-blocker dose on outcome and the differences in outcome between carvedilol and metoprolol tartrate in patients with chronic heart failure: results from the COMET trial. AB - AIMS: We studied the influence of heart rate (HR), systolic blood pressure (SBP), and beta-blocker dose on outcome in the 2599 out of 3029 patients in Carvedilol Or Metoprolol European Trial (COMET) who were alive and on study drug at 4 months after randomization (time of first visit on maintenance therapy). METHODS AND RESULTS: By multivariable analysis, baseline HR, baseline SBP, and their change after 4 months were not independently related to subsequent outcome. In a multivariable analysis including clinical variables, HR above and SBP below the median value achieved at 4 months predicted subsequent increased mortality [relative risk (RR) for HR>68 b.p.m. 1.333; 95% confidence intervals (CI) 1.152 1.542; P<0.0001 and RR for SBP>120 mmHg 0.78; 95% CI 0.671-0.907; P<0.0013]. Achieving target beta-blocker dose was associated with a better outcome (RR 0.779; 95% CI 0.662-0.916; P<0.0025). The superiority of carvedilol as compared to metoprolol tartrate was maintained in a multivariable model (RR 0.767; 95% CI 0.663-0.887; P=0.0004) and there was no interaction with HR, SBP, or beta-blocker dose. CONCLUSION: Beta-blocker dose, HR, and SBP achieved during beta-blocker therapy have independent prognostic value in heart failure. None of these factors influenced the beneficial effects of carvedilol when compared with metoprolol tartrate at the pre-defined target doses used in COMET. PMID- 16040620 TI - Soft versus firm embryo transfer catheters for assisted reproduction: a systematic review and meta-analysis. AB - BACKGROUND: The true impact of the embryo transfer catheter choice on an IVF programme has not been fully examined. We therefore decided to systematically review the evidence provided in the literature so that we may evaluate a single variable in relation to a successful transfer, the firmness of the embryo transfer catheter. METHODS: An extensive computerized search was conducted for all relevant articles published as full text, or abstracts, and critically appraised. In addition, a hand search was undertaken to locate any further trials. RESULTS: A total of 23 randomized controlled trials (RCT) evaluating the types of embryo transfer catheters were identified. Only ten of these trials, including 4141 embryo transfers, compared soft versus firm embryo catheters. Pooling of the results demonstrated a statistically significantly increased chance of clinical pregnancy following embryo transfer using the soft (643/2109) versus firm (488/2032) catheters [P = 0.01; odds ratio (OR) = 1.39, 95% confidence interval (CI) = 1.08-1.79]. When only the truly RCT were analysed, the results were again still in favour of using the soft embryo transfer catheters [soft (432/1403) versus firm (330/1402)], but with a greater significance (P < 0.00001; OR = 1.49, 95% CI = 1.26-1.77). CONCLUSION: Using soft embryo transfer catheters for embryo transfer results in a significantly higher pregnancy rate as compared to firm catheters. PMID- 16040621 TI - Antibiotics for childhood community-acquired pneumonia in a region with a high prevalence of penicillin non-susceptible Streptococcus pneumoniae. PMID- 16040622 TI - Anti-metabolic activity of caspofungin against Candida albicans and Candida parapsilosis biofilms. AB - OBJECTIVES: Candidiasis can be associated with the formation of biofilms on bioprosthetic surfaces and the intrinsic resistance of Candida albicans biofilms to the most commonly used antifungal agents has been demonstrated. In this study, we report on the antifungal activity of caspofungin at two different concentrations, on C. albicans and Candida parapsilosis biofilms with different ages of maturation. METHODS: Fifteen strains of C. albicans (10 strains susceptible to fluconazole in vitro and five strains resistant to this antifungal agent) and six strains of C. parapsilosis (all were susceptible to fluconazole in vitro) were studied. The antifungal activity of caspofungin was assessed by looking for a significant inhibition of the metabolic activity of yeasts within biofilms. Biofilms of Candida were produced in vitro, on silicone catheters. RESULTS: Caspofungin used at MIC did not modify the metabolic activity of C. albicans, whatever the maturation age of the biofilms. The same concentration of caspofungin significantly reduced the metabolism (Por=1 mg/L to voriconazole and caspofungin, respectively. Overall, 40% of patients died in-hospital for an annual mortality rate of 1.2 per 100,000. CONCLUSIONS: Candida species are an important cause of invasive infection and patients with co-morbidities and extremes of age are at highest risk. Alternatives to fluconazole should be considered for initial empiric therapy in patients with severe invasive Candida species infections. PMID- 16040624 TI - Assessment of the fitness impacts on Escherichia coli of acquisition of antibiotic resistance genes encoded by different types of genetic element. AB - OBJECTIVES: Little is known of the fitness cost that antibiotic resistance exerts on wild-type bacteria, especially in their natural environments. We therefore examined the fitness costs that several antibiotic resistance elements imposed on a wild-type Escherichia coli isolate, both in the laboratory and in a pig gut colonization model. METHODS: Plasmid R46, Tn1 and Tn7 and a K42R RpsL substitution were separately introduced into E. coli 345-2 RifC, a rifampicin resistant derivative of a recent porcine isolate. The insertion site of Tn1 was determined by DNA sequencing. The fitness cost of each resistance element was assessed in vitro by pairwise growth competition and in vivo by regularly monitoring the recovery of strains from faeces for 21 days following oral inoculation of organic piglets. Each derivative of 345-2 RifC carrying a resistance element was grown in antibiotic-free broth for 200 generations and the experiments to assess fitness were repeated. RESULTS: RpsL K42R was found to impose a small fitness cost on E. coli 345-2 RifC in vitro but did not compromise survival in vivo. R46 imposed a cost both before and after laboratory passage in vitro, but only the pre-passage strain was at a disadvantage in vivo. The post passage isolate had an advantage in pigs. Acquisition of Tn7 had no impact on the fitness of E. coli 345-2 RifC. Two derivatives containing Tn1 were isolated and, in both cases, the transposon inserted into the same cryptic chromosomal sequence. Acquisition of Tn1 improved fitness of E. coli 345-2 RifC in vitro and in vivo in the case of the first derivative, but in the case of a second, independent derivative, Tn1 had a neutral effect on fitness. CONCLUSIONS: The fitness impact imposed on E. coli 345-2 RifC by carriage of antibiotic resistance elements was generally low or non-existent, suggesting that once established, resistance may be difficult to eliminate through reduction in prescribing alone. PMID- 16040625 TI - Tigecycline. AB - New antimicrobial agents are urgently needed for clinical use due to the increasing prevalence and spread of multidrug-resistant bacteria that are commonly responsible for serious and life-threatening diseases. The need to develop new agents that effectively overcome existing mechanisms of resistance displayed by bacteria resistant to currently available drugs has become paramount. Tigecycline, the first in a new class of antimicrobials, the glycylcyclines, is an analogue of minocycline with additional properties that negate most mechanisms mediating resistance to the tetracyclines. In vitro testing has revealed that tigecycline has activity against vancomycin-resistant enterococci, methicillin-resistant Staphylococcus aureus, penicillin-resistant Streptococcus pneumoniae and many species of multidrug-resistant Gram-negative bacteria, although resistance to tigecycline by Pseudomonas aeruginosa and reduced susceptibility among Proteus species do occur. Tigecycline is being evaluated in multicentre Phase III clinical trials for therapy of many serious and life-threatening infections in which multidrug-resistant bacterial organisms may be found. Tigecycline appears to hold promise as a novel expanded spectrum antibiotic. PMID- 16040626 TI - Endothelin-mediated increases in lung VEGF content promote vascular leak in young rats exposed to viral infection and hypoxia. AB - Viral respiratory infections increase the susceptibility of young animals to hypoxia-induced pulmonary edema formation. Previous work has shown that increased lung levels of endothelin (ET) contribute to this effect, though the mechanisms by which ET promotes vascular leak remain uncertain. Both in vitro and in vivo evidence suggests that ET can upregulate the production of VEGF, which is known to increase vascular permeability. We hypothesized that increases in lung ET promote increases in lung VEGF, which in turn increases vascular leak in the lung. Weanling rats were exposed to moderate hypoxia for 24 h while recovering from a mild viral respiratory infection, to hypoxia alone, or to viral infection alone. Lung VEGF mRNA and protein content were measured by RT-PCR and Western blotting, respectively. Animals exposed to hypoxia + virus demonstrated significant increases in lung VEGF mRNA and protein content. Immunohistochemical studies showed increased VEGF expression in alveolar septa and small pulmonary vessels in those animals. ET receptor blockade with bosentan prevented this increase in lung VEGF content, suggesting that ET promotes VEGF accumulation in the lung in this setting. Animals exposed to hypoxia + virus also demonstrated substantial increases in lung albumin extravasation, and those increases were blocked by both ET receptor blockade and VEGF antagonism. These findings suggest that ET-driven increases in lung VEGF content can contribute to the formation of pulmonary edema. PMID- 16040627 TI - Regulation of constitutive neutrophil apoptosis by the alpha,beta-unsaturated aldehydes acrolein and 4-hydroxynonenal. AB - Reactive alpha,beta-unsaturated aldehydes are major components of common environmental pollutants and are products of lipid oxidation. Although these aldehydes have been demonstrated to induce apoptotic cell death in various cell types, we recently observed that the alpha,beta-unsaturated aldehyde acrolein (ACR) can inhibit constitutive apoptosis of polymorphonuclear neutrophils and thus potentially contribute to chronic inflammation. The present study was designed to investigate the biochemical mechanisms by which two representative alpha,beta-unsaturated aldehydes, ACR and 4-hydroxynonenal (HNE), regulate neutrophil apoptosis. Whereas low concentrations of either aldehyde (<10 microM) mildly promoted apoptosis in neutrophils (reflected by increased phosphatidylserine exposure, caspase-3 activation, and mitochondrial cytochrome c release), higher concentrations prevented critical features of apoptosis (caspase 3 activation, phosphatidylserine exposure) and caused delayed neutrophil cell death with characteristics of necrosis/oncosis. Inhibition of caspase-3 activation by either aldehyde occurred despite increases in mitochondrial cytochrome c release and occurred in close association with depletion of cellular GSH and with cysteine modifications within caspase-3. However, procaspase-3 processing was also prevented, because of inhibited activation of caspases-9 and 8 under similar conditions, suggesting that ACR (and to a lesser extent HNE) can inhibit both intrinsic (mitochondria dependent) and extrinsic mechanisms of neutrophil apoptosis at initial stages. Collectively, our results indicate that alpha,beta-unsaturated aldehydes can inhibit constitutive neutrophil apoptosis by common mechanisms, involving changes in cellular GSH status resulting in reduced activation of initiator caspases as well as inactivation of caspase-3 by modification of its critical cysteine residue. PMID- 16040628 TI - Regulation of reactive oxygen species-induced endothelial cell-cell and cell matrix contacts by focal adhesion kinase and adherens junction proteins. AB - Oxidants, generated by activated neutrophils, have been implicated in the pathophysiology of vascular disorders and lung injury; however, mechanisms of oxidant-mediated endothelial barrier dysfunction are unclear. Here, we have investigated the role of focal adhesion kinase (FAK) in regulating hydrogen peroxide (H(2)O(2))-mediated tyrosine phosphorylation of intercellular adhesion proteins and barrier function in endothelium. Treatment of bovine pulmonary artery endothelial cells (BPAECs) with H(2)O(2) increased tyrosine phosphorylation of FAK, paxillin, beta-catenin, and vascular endothelial (VE) cadherin and decreased transendothelial electrical resistance (TER), an index of cell-cell adhesion and/or cell-matrix adhesion. To study the role of FAK in H(2)O(2)-induced TER changes, BPAECs were transfected with vector or FAK wild type or FAK-related non-kinase (FRNK) plasmids. Overexpression of FRNK reduced FAK expression and attenuated H(2)O(2)-mediated tyrosine phosphorylation of FAK, paxillin, beta-catenin, and VE-cadherin and cell-cell adhesion. Additionally, FRNK prevented H(2)O(2)-induced distribution of FAK, paxillin, beta-catenin, or VE-cadherin toward focal adhesions and cell-cell adhesions but not actin stress fiber formation. These results suggest that activation of FAK by H(2)O(2) is an important event in oxidant-mediated VE barrier function regulated by cell-cell and cell-matrix contacts. PMID- 16040629 TI - Beta-catenin regulates differentiation of respiratory epithelial cells in vivo. AB - An activated form of beta-catenin [Catnb(Delta(ex3))] was expressed in respiratory epithelial cells of the developing lung. Although morphogenesis was not altered at birth, air space enlargement and epithelial cell dysplasia were observed in the early postnatal period and persisted into adulthood. The Catnb(Delta(ex3)) protein caused squamous, cuboidal, and goblet cell dysplasia in intrapulmonary conducting airways. Atypical epithelial cells that stained for surfactant pro protein C (pro-SP-C) and had morphological characteristics of alveolar type II cells were observed in bronchioles of the transgenic mice. Catnb(Delta(ex3)) inhibited expression of Foxa2 and caused goblet cell hyperplasia associated with increased staining for mucins and the MUC5A/C protein. In vitro, both wild type and activated beta-catenin negatively regulated the expression of the Foxa2 promoter. Catnb(Delta(ex3)) also caused pulmonary tumors in adult mice. Activation of beta-catenin caused ectopic differentiation of alveolar type II-like cells in conducting airways, goblet cell hyperplasia, and air space enlargement, demonstrating a critical role for the Wnt/beta-catenin signal transduction pathway in the differentiation of the respiratory epithelium in the postnatal lung. PMID- 16040630 TI - Sensitization of pulmonary chemosensitive neurons by bombesin-like peptides in rats. AB - Small cell lung cancer (SCLC) patients suffer from pulmonary stresses such as dyspnea and chest pain, and the pathogenic mechanisms are not known. SCLC cells secrete a variety of bioactive neuropeptides, including bombesin-like peptides. We hypothesize that these peptides may enhance the sensitivity of the pulmonary chemosensitive nerve endings, contributing to the development of these pulmonary stresses in SCLC patients. This study was therefore carried out to determine the effects of bombesin and gastrin-releasing peptide (GRP), a major bombesin-like peptide, on the sensitivities of pulmonary chemoreflex and isolated pulmonary vagal chemosensitive neurons. In anesthetized, spontaneously breathing rats, intravenous infusion of bombesin or GRP significantly amplified the pulmonary chemoreflex responses to chemical stimulants such as capsaicin and ATP. The enhanced responses were completely abolished by perineural capsaicin treatment of both cervical vagi, suggesting the involvement of pulmonary C-fiber afferents. In isolated pulmonary vagal chemosensitive neurons, pretreatment with bombesin or GRP potentiated the capsaicin-induced Ca(2+) transient. This sensitizing effect was further demonstrated in patch-clamp recording studies; the sensitivities of these neurons to both chemical (capsaicin and ATP) and electrical stimuli were significantly enhanced by the presence of either bombesin or GRP. In summary, our results have demonstrated that bombesin and GRP upregulate the pulmonary chemoreflex sensitivity in vivo and the excitability of isolated pulmonary chemosensitive neurons in vitro. PMID- 16040631 TI - Effects of rottlerin on silica-exacerbated systemic autoimmune disease in New Zealand mixed mice. AB - Environmental crystalline silica exposure has been associated with formation of autoantibodies and development of systemic autoimmune disease, but the mechanisms leading to these events are unknown. Silica exposure in autoimmune-prone New Zealand mixed (NZM) mice results in a significant exacerbation of systemic autoimmunity as measured by increases in autoantibodies and glomerulonephritis. Previous studies have suggested that silica-induced apoptosis of alveolar macrophages (AM) contributes to the generation of the autoantibodies and disease. Rottlerin has been reported to inhibit apoptosis in many cell types, possibly through direct or indirect effects on PKCdelta. In this study, rottlerin reduced silica-induced apoptosis in bone marrow-derived macrophages as measured by DNA fragmentation. In NZM mice, RNA and protein levels of PKCdelta were significantly elevated in AM 14 wk after silica exposure. Therefore, rottlerin was used to reduce apoptosis of AM and evaluate the progress of silica-exacerbated systemic autoimmune disease. Fourteen weeks after silica exposure, NZM mice had increased levels of anti-histone autoantibodies, high proteinuria, and glomerulonephritis. However, silica-instilled mice that also received weekly instillations of rottlerin had significantly lower levels of proteinuria, anti-histone autoantibodies, complement C3, and IgG deposition within the kidney. Weekly instillations of rottlerin in silica-instilled NZM mice also inhibited the upregulation of PKCdelta in AM. Together, these data demonstrate that in vivo treatment with rottlerin significantly decreased the exacerbation of autoimmunity by silica exposure. PMID- 16040632 TI - Acetylcholine-induced phosphorylation and membrane translocation of CPI-17 in bronchial smooth muscle of rats. AB - A translocation of protein kinase C (PKC) from cytosol to plasma membrane has been reported as an association with agonist-induced Ca2+ sensitization in smooth muscle contraction. Therefore, it is possible that a downstream target of PKC, CPI-17 [PKC-potentiated inhibitory protein for heterotrimeric myosin light chain (MLC) phosphatase of 17 kDa], might also be translocated to membrane when activated. To confirm this hypothesis, cytosolic and membrane CPI-17 was measured in acetylcholine (ACh)- and high-K+ depolarization-stimulated bronchial smooth muscle of rats. An active form of CPI-17, i.e., Thr38-phosphorylated CPI-17, was also measured in cytosolic and membrane fractions. Immunoblot analyses demonstrated a translocation of CPI-17 from cytosolic to membrane fraction by ACh, but not high-K+ depolarization, stimulation in time- and concentration dependent manners. Interestingly, phosphorylated CPI-17 was detected only in membrane fractions in the ACh-stimulated tissues. However, in the high-K+ depolarization-stimulated tissues, phosphorylated CPI-17 was not detected both in membrane and cytosolic fraction. To estimate downstream of activated CPI-17, immunoblotting for phosphorylated MLC was performed in ACh- or high-K+ depolarization-stimulated tissues. ACh- and high-K+ depolarization-induced phosphorylation of MLC was observed in its contraction-dependent manner. In conclusion, we, for the first time, suggested that CPI-17 is translocated and phosphorylated by ACh, but not high-K+ depolarization, in rat bronchial smooth muscle. ACh-induced translocation and phosphorylation of CPI-17 might be caused via the activation of muscarinic receptor. PMID- 16040633 TI - Arabidopsis SENESCENCE-ASSOCIATED GENE101 stabilizes and signals within an ENHANCED DISEASE SUSCEPTIBILITY1 complex in plant innate immunity. AB - Plant innate immunity against invasive biotrophic pathogens depends on the intracellular defense regulator ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1). We show here that Arabidopsis thaliana EDS1 interacts in vivo with another protein, SENESCENCE-ASSOCIATED GENE101 (SAG101), discovered through a proteomic approach to identify new EDS1 pathway components. Together with PHYTOALEXIN-DEFICIENT4 (PAD4), a known EDS1 interactor, SAG101 contributes intrinsic and indispensable signaling activity to EDS1-dependent resistance. The combined activities of SAG101 and PAD4 are necessary for programmed cell death triggered by the Toll Interleukin-1 Receptor type of nucleotide binding/leucine-rich repeat immune receptor in response to avirulent pathogen isolates and in restricting the growth of normally virulent pathogens. We further demonstrate by a combination of cell fractionation, coimmunoprecipitation, and fluorescence resonance energy transfer experiments the existence of an EDS1-SAG101 complex inside the nucleus that is molecularly and spatially distinct from EDS1-PAD4 associations in the nucleus and cytoplasm. By contrast, EDS1 homomeric interactions were detected in the cytoplasm but not inside the nucleus. These data, combined with evidence for coregulation between individual EDS1 complexes, suggest that dynamic interactions of EDS1 and its signaling partners in multiple cell compartments are important for plant defense signal relay. PMID- 16040634 TI - Splenic rupture and haemoperitoneum in a patient with non-compaction of the left ventricular myocardium. AB - The anaesthetic and critical care management of blunt abdominal trauma in a patient previously diagnosed with non-compaction of the left ventricular myocardium (a rare autosomal dominant inherited disease) is reported. The management was influenced by the presence of an implanted automated internal defibrillator and treatment with anticoagulants because of the high frequency of severe arrhythmias and systemic embolism. The pathophysiology of ventricular non compaction is reviewed briefly. PMID- 16040635 TI - Clinical actions of subarachnoid sevoflurane administration in vivo: a study in dogs. AB - BACKGROUND: Halogenated ethers produce clinical effects at spinal sites. Nevertheless, in vitro and in vivo studies have not determined whether the immobilizing effect in the spinal cord is due to inhibition of nociceptive or motor transmission or both. Our goal was to characterize the clinical effects of direct spinal sevoflurane administration. METHODS: Five adult beagle dogs completed the study. In a randomized and blinded manner each animal received placebo (saline 0.1 ml kg(-1)) and three concentrations of pure sevoflurane administered intrathecally (0.05, 0.075 and 0.1 ml kg(-1)) by means of a permanent spinal catheter. Sensory and motor block and state of consciousness were determined at baseline and at predetermined regular intervals until at least 2 h after total recovery. RESULTS: None of the dogs presented a decrease in consciousness with either 0.05 or 0.075 ml kg(-1) of sevoflurane. Administration of 0.1 ml kg(-1) produced light sedation (2 on a four-point sedation scale) in three of the five dogs. A comparison of the duration of the sensory and motor blocks among the three sevoflurane dosages shows a significant dose-dependent increase that is greater in all cases than that for the saline solution. CONCLUSIONS: Spinal administration of pure sevoflurane resulted in a dose-related and totally reversible motor and sensory regional block without any signs of clinical neurotoxicity or significant decrease in consciousness. Therefore the model allows us to comment on the analgesic effects at the spinal level in addition to the direct immobilizing effects of sevoflurane. PMID- 16040636 TI - Isoflurane does not mimic ischaemic preconditioning in decreasing hydroxyl radical production in the rabbit. AB - BACKGROUND: Reactive oxygen species are an important mediator in isoflurane induced myocardial preconditioning. However, hydroxyl radicals are also released during reperfusion after regional ischaemia. The purpose of the present study was to test whether ischaemic preconditioning and isoflurane would influence the production of hydroxyl radicals during reperfusion. METHODS: After i.v. administration of salicylate 100 mg kg(-1) and a 30 min stabilization period, New Zealand White rabbits were subjected to 40 min of regional myocardial ischaemia and 2 h of reperfusion. Ischaemic preconditioning was elicited by 5 min ischaemia followed by 10 min reperfusion (before the 40 min ischaemia). In another group, isoflurane (2.1%) was administered for 30 min, followed by 15 min washout, before the long ischaemia. Area at risk and infarct size were assessed by blue dye injection and tetrazolium chloride staining. We quantified the level of OH mediated conversion of salicylate to its dihydrobenzoate derivatives (2,3- and 2,5-DHBAs). Normalized values of the DHBAs (ng DHBA per mg salicylate) were calculated. RESULTS: Mean (se) infarct size was 57 (6)% of the risk area in the untreated controls. This was significantly smaller in the ischaemic preconditioning and isoflurane groups: 22 (5) and 23 (6)% respectively. At 10 min of reperfusion, ischaemic preconditioning limited the mean increase in 2,3-DHBA to 24% from baseline, compared with 81% in control and 74% in the isoflurane group. Normalized 2,5-DHBA was maximally increased by 75% in the untreated group, 4 min after reperfusion. Ischaemic preconditioning significantly inhibited this increase (24% increase from baseline, P<0.01). However, the increase observed in the isoflurane group was not different from control (71%). CONCLUSIONS: As already known, ischaemic preconditioning and isoflurane markedly reduced infarct size. However, only ischaemic preconditioning decreased postischaemic production of hydroxyl radicals. These different effects suggest different protective mechanisms at the cellular level. PMID- 16040637 TI - Effect of epidural volume extension on dose requirement of intrathecal hyperbaric bupivacaine at Caesarean section. AB - BACKGROUND: The technique of epidural volume extension (EVE) involves the injection of saline into the extradural space immediately following the intrathecal injection, as part of a combined spinal-epidural (CSE) anaesthetic. One of the suggested benefits of EVE is a reduction in local anaesthetic required. The aim of this study was to test this hypothesis by comparing the median effective doses (ED50) of hyperbaric bupivacaine with fentanyl 25 microg with and without EVE for Caesarean section. METHODS: Sixty women were randomized to receive either CSE anaesthesia with EVE (EVE group) or no EVE (NEVE group). Using a double-blinded, up-down sequential technique, varying doses of bupivacaine with fentanyl 25 microg were administered. ED50 was estimated from up down reversals and probit regression. RESULTS: The ED50 for bupivacaine was similar and not significantly different in the two groups (5.1 mg in the EVE and 6.1 mg in the NEVE group; difference 1.0 mg, 95% CI -0.12 to 2.2, P=0.08). CONCLUSIONS: This study illustrates that whilst low doses of intrathecal bupivacaine can be effectively used for Caesarean section, at such doses EVE does not appear to offer reliable or clinically relevant reductions in dosing with intrathecal bupivacaine. PMID- 16040638 TI - Usefulness of the carina as a radiographic landmark for central venous catheter placement in paediatric patients. AB - BACKGROUND: Several reports have proposed radiographic landmarks for the proper positioning of central venous catheters (CVC). The carina is one of the proposed landmarks in adults. Here, we evaluate the possibility of using the carina as a radiographic landmark for the identification of proper positioning of the CVC tip in paediatric patients. METHODS: We studied 57 right internal jugular vein catheterizations in infants and children undergoing surgery for the treatment of congenital heart disease. After placing the CVC tip at the junction of the superior vena cava and the right atrium (SVC-RA junction) via intraoperative transoesophageal echocardiography, and by taking postoperative anterior-posterior chest radiographs, we measured the longitudinal distance from the carina to the SVC-RA junction, using the Picture Archiving and Communicating System. RESULTS: The average distance between the carina and the SVC-RA junction was 1.5 cm (95% CI 1.3-1.8 cm). No catheter tip was above the carina. Although there was no particular relationship between this distance and the patient's age, height, or weight, the distance between the carina and the SVC-RA junction tended to be more variable in younger and smaller children. CONCLUSIONS: The carina can be used as a radiographic landmark for the proper CVC tip placement in paediatric patients. If the tip of the CVC is not distal to the carina the chances are minute that it is in the right atrium. PMID- 16040639 TI - Rigid bronchoscopy-guided percutaneous tracheostomy. AB - We describe a case series of seven patients that demonstrates the usefulness of rigid bronchoscopy in percutaneous tracheostomy. The technique was used in selected patients who had a previous tracheostomy, a difficult airway, high risk of bleeding, or a tracheal stent in place. PMID- 16040645 TI - Post-Lyme borreliosis syndrome: a meta-analysis of reported symptoms. AB - BACKGROUND: This meta-analysis compares the prevalence of fatigue, musculoskeletal pain, and neurocognitive difficulties in patients who have had Lyme borreliosis (LB) and control subjects without LB. METHODS: Titles and abstracts in PubMed were reviewed for studies with data on the symptoms listed above that compared patients who had had LB with controls from the general population. Five studies with 504 patients and 530 controls were included in the meta-analysis. RESULTS: The prevalence of symptoms was significantly higher in the LB patients, with P-values between <0.00001 and 0.007 for 8 of the 10 symptoms in the three categories listed above. The higher prevalence of certain neurocognitive symptoms but not others, in the same pattern as reported in the literature, is further confirmation of this syndrome. The pattern of symptoms appears to be different from that seen in fibromyalgia, depression, and chronic fatigue syndrome. CONCLUSIONS: This meta-analysis provides strong evidence that some patients with LB have fatigue, musculoskeletal pain, and neurocognitive difficulties that may last for years despite antibiotic treatment. PMID- 16040646 TI - The tobacco mosaic virus 126-kilodalton protein, a constituent of the virus replication complex, alone or within the complex aligns with and traffics along microfilaments. AB - Virus-induced cytoplasmic inclusion bodies (referred to as virus replication complexes [VRCs]) consisting of virus and host components are observed in plant cells infected with tobacco mosaic virus, but the components that modulate their form and function are not fully understood. Here, we show that the tobacco mosaic virus 126-kD protein fused with green fluorescent protein formed cytoplasmic bodies (126-bodies) in the absence of other viral components. Using mutant 126 kD:green fluorescent fusion proteins and viral constructs expressing the corresponding mutant 126-kD proteins, it was determined that the size of the 126 bodies and the corresponding VRCs changed in synchrony for each 126-kD protein mutation tested. Through colabeling experiments, we observed the coalignment and intracellular trafficking of 126-bodies and, regardless of size, VRCs, along microfilaments (MFs). Disruption of MFs with MF-depolymerizing agents or through virus-induced gene silencing compromised the intracellular trafficking of the 126 bodies and VRCs and virus cell-to-cell movement, but did not decrease virus accumulation to levels that would affect virus movement or prevent VRC formation. Our results indicate that (1) the 126-kD protein modulates VRC size and traffics along MFs in cells; (2) VRCs traffic along MFs in cells, possibly through an interaction with the 126-kD protein, and the negative effect of MF antagonists on 126-body and VRC intracellular movement and virus cell-to-cell movement correlates with the disruption of this association; and (3) virus movement was not correlated with VRC size. PMID- 16040647 TI - Arabidopsis ARP7 is an essential actin-related protein required for normal embryogenesis, plant architecture, and floral organ abscission. AB - The actin-related proteins (ARPs) that are localized to the nucleus are present as components of various chromatin-modifying complexes involved in chromatin dynamics and transcriptional regulation. Arabidopsis (Arabidopsis thaliana) ARP7 is a constitutively expressed nuclear protein belonging to a novel plant-specific ARP class. In this study, we demonstrate a vital role for ARP7 protein in embryogenesis and plant development. Knocking out the expression of ARP7 in an arp7-1 T-DNA mutant produced morphologically aberrant, homozygous embryos that were arrested at or before the torpedo stage of development. Hence, the arp7-1 null mutation is homozygous lethal. Knocking down the expression levels of ARP7 protein with RNA interference produced viable Arabidopsis lines affected in multiple developmental pathways and induced dosage-dependent, heritable defects in plant architecture. The transgenic plants containing greatly reduced levels of ARP7 in the nucleus were severely dwarfed with small rosette leaves that are defective in cell expansion and trichome morphology. Moreover, the ARP7-deficient RNA interference plants exhibited retarded root growth, altered flower development, delayed perianth abscission, and reduced fertility. These pleiotropic phenotypic changes suggest a critical role for the Arabidopsis ARP7 protein in the regulation of various phases of plant development through chromatin-mediated, global regulation of gene expression. PMID- 16040648 TI - Characteristics of electrical signals in poplar and responses in photosynthesis. AB - To gain an understanding of the role of electrical signaling in trees, poplar (Populus trichocarpa, Populus tremula x P. tremuloides) shoots were stimulated by chilling as well as flaming. Two kinds of signal propagation were detected by microelectrode measurements (aphid technique) in the phloem of leaf veins: (1) basipetal, short-distance signaling that led to rapid membrane hyperpolarization caused by K+-efflux within the leaf lamina; and (2) acropetal, long-distance signaling that triggered depolarization of the membrane potential in the leaf phloem. In the latter, the depolarizing signals travel across the stem from the manipulated leaves to adjacent leaves where the net CO2 uptake rate is temporarily depressed toward compensation. With regard to photosystem II, both heat-induced long-distance and short-distance signaling were investigated using two-dimensional "imaging" analysis of chlorophyll fluorescence. Both types of signaling significantly reduced the quantum yield of electron transport through photosystem II. Imaging analysis revealed that the signal that causes yield reduction spreads through the leaf lamina. Coldblocking of the stem proved that the electrical signal transmission via the phloem becomes disrupted, causing the leaf gas exchange to remain unaffected. Calcium-deficient trees showed a marked contrast inasmuch as the amplitude of the electrical signal was distinctly reduced, concomitant with the absence of a significant response in leaf gas exchange upon flame wounding. In summary, the above results led us to conclude that calcium as well as potassium is involved in the propagation of phloem transmitted electrical signals that evoke specific responses in the photosynthesis of leaves. PMID- 16040649 TI - Sites and regulation of polyamine catabolism in the tobacco plant. Correlations with cell division/expansion, cell cycle progression, and vascular development. AB - We previously gave a picture of the homeostatic characteristics of polyamine (PA) biosynthesis and conjugation in tobacco (Nicotiana tabacum) plant organs during development. In this work, we present the sites and regulation of PA catabolism related to cell division/expansion, cell cycle progression, and vascular development in the tobacco plant. Diamine oxidase (DAO), PA oxidase (PAO), peroxidases (POXs), and putrescine N-methyltransferase expressions follow temporally and spatially discrete patterns in shoot apical cells, leaves (apical, peripheral, and central regions), acropetal and basipetal petiole regions, internodes, and young and old roots in developing plants. DAO and PAO produce hydrogen peroxide, a plant signal molecule and substrate for POXs. Gene expression and immunohistochemistry analyses reveal that amine oxidases in developing tobacco tissues precede and overlap with nascent nuclear DNA and also with POXs and lignification. In mature and old tissues, flow cytometry indicates that amine oxidase and POX activities, as well as pao gene and PAO protein levels, coincide with G2 nuclear phase and endoreduplication. In young versus the older roots, amine oxidases and POX expression decrease with parallel inhibition of G2 advance and endoreduplication, whereas putrescine N-methyltransferase dramatically increases. In both hypergeous and hypogeous tissues, DAO and PAO expression occurs in cells destined to undergo lignification, suggesting a different in situ localization. DNA synthesis early in development and the advance in cell cycle/endocycle are temporally and spatially related to PA catabolism and vascular development. PMID- 16040650 TI - Fractionation of the three stable oxygen isotopes by oxygen-producing and oxygen consuming reactions in photosynthetic organisms. AB - The triple isotope composition (delta17O and delta18O) of dissolved O2 in the ocean and in ice cores was recently used to assess the primary productivity over broad spatial and temporal scales. However, assessment of the productivity with the aid of this method must rely on accurate measurements of the 17O/16O versus 18O/16O relationship in each of the main oxygen-producing and -consuming reactions. Data obtained here showed that cleavage of water in photosystem II did not fractionate oxygen isotopes; the delta18O and delta17O of the O2 evolved were essentially identical to those of the substrate water. The fractionation slopes for the oxygenase reaction of Rubisco and respiration were identical (0.518 +/- 0.001) and that of glycolate oxidation was 0.503 +/- 0.002. There was a considerable difference in the slopes of O2 photoreduction (the Mehler reaction) in the cyanobacterium Synechocystis sp. strain PCC 6803 (0.497 +/- 0.004) and that of pea (Pisum sativum) thylakoids (0.526 +/- 0.001). These values provided clear and independent evidence that the mechanism of O2 photoreduction differs between higher plants and cyanobacteria. We used our method to assess the magnitude of O2 photoreduction in cyanobacterial cells maintained under conditions where photorespiration was negligible. It was found that electron flow to O2 can be as high as 40% that leaving photosystem II, whereas respiratory activity in the light is only 6%. The implications of our findings to the evaluation of specific O2-producing or -consuming reactions, in vivo, are discussed. PMID- 16040651 TI - An RNA-dependent RNA polymerase prevents meristem invasion by potato virus X and is required for the activity but not the production of a systemic silencing signal. AB - One of the functions of RNA silencing in plants is antiviral defense. A hallmark of RNA silencing is spreading of the silenced state through the plant. Little is known about the nature of the systemic silencing signal and the proteins required for its production, transport, and reception in plant tissues. Here, we show that the RNA-dependent RNA polymerase RDR6 in Nicotiana benthamiana is involved in defense against potato virus X at the level of systemic spreading and in exclusion of the virus from the apical growing point. It has no effect on primary replication and cell-to-cell movement of the virus and does not contribute significantly to the formation of virus-derived small interfering (si) RNA in a fully established potato virus X infection. In grafting experiments, the RDR6 homolog was required for the ability of a cell to respond to, but not to produce or translocate, the systemic silencing signal. Taking these findings together, we suggest a model of virus defense in which RDR6 uses incoming silencing signal to generate double-stranded RNA precursors of secondary siRNA. According to this idea, the secondary siRNAs mediate RNA silencing as an immediate response that slows down the systemic spreading of the virus into the growing point and newly emerging leaves. PMID- 16040652 TI - Analysis of the rice mutant dwarf and gladius leaf 1. Aberrant katanin-mediated microtubule organization causes up-regulation of gibberellin biosynthetic genes independently of gibberellin signaling. AB - Molecular genetic studies of plant dwarf mutants have indicated that gibberellin (GA) and brassinosteroid (BR) are two major factors that determine plant height; dwarf mutants that are caused by other defects are relatively rare, especially in monocot species. Here, we report a rice (Oryza sativa) dwarf mutant, dwarf and gladius leaf 1 (dgl1), which exhibits only minimal response to GA and BR. In addition to the dwarf phenotype, dgl1 produces leaves with abnormally rounded tip regions. Positional cloning of DGL1 revealed that it encodes a 60-kD microtubule severing katanin-like protein. The protein was found to be important in cell elongation and division, based on the observed cell phenotypes. GA biosynthetic genes are up-regulated in dgl1, but the expression of BR biosynthetic genes is not enhanced. The enhanced expression of GA biosynthetic genes in dgl1 is not caused by inappropriate GA signaling because the expression of these genes was repressed by GA3 treatment, and degradation of the rice DELLA protein SLR1 was triggered by GA3 in this mutant. Instead, aberrant microtubule organization caused by the loss of the microtubule-severing function of DGL1 may result in enhanced expression of GA biosynthetic genes in that enhanced expression was also observed in a BR-deficient mutant with aberrant microtubule organization. These results suggest that the function of DGL1 is important for cell and organ elongation in rice, and aberrant DGL1-mediated microtubule organization causes up regulation of gibberellin biosynthetic genes independently of gibberellin signaling. PMID- 16040653 TI - Expression of Arabidopsis MIRNA genes. AB - MicroRNAs (miRNAs) are approximately 21-nucleotide noncoding RNAs that regulate target transcripts in plants and animals. In addition to miRNAs, plants contain several classes of endogenous small interfering RNAs (siRNAs) involved in target gene regulation and epigenetic silencing. Small RNA libraries were constructed from wild-type Arabidopsis (Arabidopsis thaliana) and mutant plants (rdr2 and dcl3) that were genetically enriched for miRNAs, and a computational procedure was developed to identify candidate miRNAs. Thirty-eight distinct miRNAs corresponding to 22 families were represented in the libraries. Using a 5' rapid amplification of cDNA ends procedure, the transcription start sites for 63 miRNA primary transcripts from 52 MIRNA loci (99 loci tested) were mapped, revealing features consistent with an RNA polymerase II mechanism of transcription. Ten loci (19%) yielded transcripts from multiple start sites. A canonical TATA box motif was identified upstream of the major start site at 45 (86%) of the mapped MIRNA loci. The 5'-mapping data were combined with miRNA cloning and 3'-PCR data to definitively validate expression of at least 73 MIRNA genes. These data provide a molecular basis to explore regulatory mechanisms of miRNA expression in plants. PMID- 16040654 TI - Structure-based in vitro engineering of the anthranilate synthase, a metabolic key enzyme in the plant tryptophan pathway. AB - Rice (Oryza sativa) anthranilate synthase alpha-subunit, OASA2, was modified by in vitro mutagenesis based on structural information from bacterial homologs. Twenty-four amino acid residues, predicted as putative tryptophan binding sites or their proximal regions in the OASA2 sequence, were selected and 36 mutant OASA2 genes were constructed by PCR-based site-directed mutagenesis. Corresponding mutant proteins were synthesized in a combination of two in vitro systems, transcription with a bacteriophage SP6 RNA polymerase and translation with a wheat-embryo cell-free system. Enzymatic functions of the mutant proteins were simultaneously examined, and we found six mutants with elevated catalytic activity and five mutants with enhanced tolerance to feedback inhibition by tryptophan. Moreover, we observed that some sets of specific combinations of the novel mutations additively conferred both characteristics to the mutant enzymes. The functions of the mutant enzymes were confirmed in vivo. The free tryptophan content of mutant rice calli expressing OASA2 enzyme with a double mutation was 30-fold of that of untransformed calli. Thus, our in vitro approach utilizing structural information of bacterial homologs is a potent technique to generate designer enzymes with predefined functions. PMID- 16040655 TI - Adaptations required for mitochondrial import following mitochondrial to nucleus gene transfer of ribosomal protein S10. AB - The minimal requirements to support protein import into mitochondria were investigated in the context of the phenomenon of ongoing gene transfer from the mitochondrion to the nucleus in plants. Ribosomal protein 10 of the small subunit is encoded in the mitochondrion in soybean and many other angiosperms, whereas in several other species it is nuclear encoded and thus must be imported into the mitochondrial matrix to function. When encoded by the nuclear genome, it has adopted different strategies for mitochondrial targeting and import. In lettuce (Lactuca sativa) and carrot (Daucus carota), Rps10 independently gained different N-terminal extensions from other genes, following transfer to the nucleus. (The designation of Rps10 follows the following convention. The gene is indicated in italics. If encoded in the mitochondrion, it is rps10; if encoded in the nucleus, it is Rps10.) Here, we show that the N-terminal extensions of Rps10 in lettuce and carrot are both essential for mitochondrial import. In maize (Zea mays), Rps10 has not acquired an extension upon transfer but can be readily imported into mitochondria. Deletion analysis located the mitochondrial targeting region to the first 20 amino acids. Using site directed mutagenesis, we changed residues in the first 20 amino acids of the mitochondrial encoded soybean (Glycine max) rps10 to the corresponding amino acids in the nuclear encoded maize Rps10 until import was achieved. Changes were required that altered charge, hydrophobicity, predicted ability to form an amphipathic alpha-helix, and generation of a binding motif for the outer mitochondrial membrane receptor, translocase of the outer membrane 20. In addition to defining the changes required to achieve mitochondrial localization, the results demonstrate that even proteins that do not present barriers to import can require substantial changes to acquire a mitochondrial targeting signal. PMID- 16040656 TI - Possible involvement of phototropins in leaf movement of kidney bean in response to blue light. AB - The leaf of kidney bean (Phaseolus vulgaris) moves in response to blue light. The movement is induced by a decrease in the turgor pressure of pulvinar motor cells on the irradiated side. In this study, we investigated the initial event of the movement with respect to function of phototropin and the plasma membrane H+ ATPase in the motor cells. The results indicated that, in dark conditions, phototropin existed in a dephosphorylated state and the H+-ATPase existed in a phosphorylated state. A pulse of blue light (30 s) induced the phosphorylation of phototropin and the dephosphorylation of the H+-ATPase as determined by the binding behavior of 14-3-3 protein. Phototropin phosphorylation occurred rapidly, followed by the transient gradual dephosphorylation of the H+-ATPase. When the specific flavoprotein inhibitor diphenyleneiodonium and the protein kinase inhibitors K-252a and staurosporine were administered to pulvinar cells, both phototropin phosphorylation and H+-ATPase dephosphorylation were inhibited. The phosphorylation and dephosphorylation exhibited similar fluence rate dependencies to blue light. These results indicated that phototropin may function upstream of the plasma membrane H+-ATPase and decrease the activity of H+-ATPase by dephosphorylation. We provide evidence for the existence of three kinds of phototropins in pulvinar motor cells. PMID- 16040657 TI - Arabidopsis cyp51 mutant shows postembryonic seedling lethality associated with lack of membrane integrity. AB - CYP51 exists in all organisms that synthesize sterols de novo. Plant CYP51 encodes an obtusifoliol 14alpha-demethylase involved in the postsqualene sterol biosynthetic pathway. According to the current gene annotation, the Arabidopsis (Arabidopsis thaliana) genome contains two putative CYP51 genes, CYP51A1 and CYP51A2. Our studies revealed that CYP51A1 should be considered an expressed pseudogene. To study the functional importance of the CYP51A2 gene in plant growth and development, we isolated T-DNA knockout alleles for CYP51A2. Loss-of function mutants for CYP51A2 showed multiple defects, such as stunted hypocotyls, short roots, reduced cell elongation, and seedling lethality. In contrast to other sterol mutants, such as fk/hydra2 and hydra1, the cyp51A2 mutant has only minor defects in early embryogenesis. Measurements of endogenous sterol levels in the cyp51A2 mutant revealed that it accumulates obtusifoliol, the substrate of CYP51, and a high proportion of 14alpha-methyl-delta8-sterols, at the expense of campesterol and sitosterol. The cyp51A2 mutants have defects in membrane integrity and hypocotyl elongation. The defect in hypocotyl elongation was not rescued by the exogenous application of brassinolide, although the brassinosteroid-signaling cascade is apparently not affected in the mutants. Developmental defects in the cyp51A2 mutant were completely rescued by the ectopic expression of CYP51A2. Taken together, our results demonstrate that the Arabidopsis CYP51A2 gene encodes a functional obtusifoliol 14alpha-demethylase enzyme and plays an essential role in controlling plant growth and development by a sterol-specific pathway. PMID- 16040658 TI - Arabidopsis peroxin 16 coexists at steady state in peroxisomes and endoplasmic reticulum. AB - Homologs of peroxin 16 genes (PEX16) have been identified only in Yarrowia lipolytica, humans (Homo sapiens), and Arabidopsis (Arabidopsis thaliana). The Arabidopsis gene (AtPEX16), previously reported as the SSE1 gene, codes for a predicted 42-kD membrane peroxin protein (AtPex16p). Lin et al. (Y. Lin, J.E. Cluette-Brown, H.M. Goodman [2004] Plant Physiol 135: 814-827) reported that SSE1/AtPEX16 was essential for endoplasmic reticulum (ER)-dependent oil and protein body biogenesis in peroxisome-deficient maturing seeds and likely also was involved in peroxisomal biogenesis based on localization of stably expressed green fluorescent protein::AtPex16p in peroxisomes of Arabidopsis plants. In this study with Arabidopsis suspension-cultured cells, combined in vivo and in vitro experiments revealed a novel dual organelle localization and corresponding membrane association/topology of endogenous AtPex16p. Immunofluorescence microscopy with antigen affinity-purified IgGs showed an unambiguous, steady state coexistence of AtPex16p in suspension cell peroxisomes and ER. AtPex16p also was observed in peroxisomes and ER of root and leaf cells. Cell fractionation experiments surprisingly revealed two immunorelated polypeptides, 42 kD (expected) and 52 kD (unexpected), in homogenates and microsome membrane pellets derived from roots, inflorescence, and suspension cells. Suc-gradient purifications confirmed the presence of both 42-kD and 52-kD polypeptides in isolated peroxisomes (isopycnic separation) and in rough ER vesicles (Mg2+ shifted). They were found peripherally associated with peroxisome and ER membranes but not as covalently bound subunits of AtPex16p. Both were mostly on the matrix side of peroxisomal membranes and unexpectedly mostly on the cytosolic side of ER membranes. In summary, AtPex16p is the only authentic plant peroxin homolog known to coexist at steady state within peroxisomes and ER; these data provide new insights in support of its ER-related, multifunctional roles in organelle biogenesis. PMID- 16040659 TI - Autophagic nutrient recycling in Arabidopsis directed by the ATG8 and ATG12 conjugation pathways. AB - Autophagy is an important mechanism for nonselective intracellular breakdown whereby cytosol and organelles are encapsulated in vesicles, which are then engulfed and digested by lytic vacuoles/lysosomes. In yeast, this encapsulation employs a set of autophagy (ATG) proteins that direct the conjugation of two ubiquitin-like protein tags, ATG8 and ATG12, to phosphatidylethanolamine and the ATG5 protein, respectively. Using an Arabidopsis (Arabidopsis thaliana) atg7 mutant unable to ligate either tag, we previously showed that the ATG8/12 conjugation system is important for survival under nitrogen-limiting growth conditions. By reverse-genetic analyses of the single Arabidopsis gene encoding ATG5, we show here that the subpathway that forms the ATG12-ATG5 conjugate also has an essential role in plant nutrient recycling. Similar to plants missing ATG7, those missing ATG5 display early senescence and are hypersensitive to either nitrogen or carbon starvation, which is accompanied by a more rapid loss of organellar and cytoplasmic proteins. Multiple ATG8 isoforms could be detected immunologically in seedling extracts. Their abundance was substantially elevated in both the atg5 and atg7 mutants, caused in part by an increase in abundance of several ATG8 mRNAs. Using a green fluorescent protein-ATG8a fusion in combination with concanamycin A, we also detected the accumulation of autophagic bodies inside the vacuole. This accumulation was substantially enhanced by starvation but blocked in the atg7 background. The use of this fusion in conjunction with atg mutants now provides an important marker to track autophagic vesicles in planta. PMID- 16040660 TI - A role for auxin redistribution in the responses of the root system architecture to phosphate starvation in Arabidopsis. AB - The changes in root system architecture (RSA) triggered by phosphate (P) deprivation were studied in Arabidopsis (Arabidopsis thaliana) plants grown for 14 d on 1 mM or 3 microM P. Two different temporal phases were observed in the response of RSA to low P. First, lateral root (LR) development was promoted between days 7 and 11 after germination, but, after day 11, all root growth parameters were negatively affected, leading to a general reduction of primary root (PR) and LR lengths and of LR density. Low P availability had contrasting effects on various stages of LR development, with a marked inhibition of primordia initiation but a strong stimulation of activation of the initiated primordia. The involvement of auxin signaling in these morphological changes was investigated in wild-type plants treated with indole-3-acetic acid or 2,3,5 triiodobenzoic acid and in axr4-1, aux1-7, and eir1-1 mutants. Most effects of low P on RSA were dramatically modified in the mutants or hormone-treated wild type plants. This shows that auxin plays a major role in the P starvation-induced changes of root development. From these data, we hypothesize that several aspects of the RSA response to low P are triggered by local modifications of auxin concentration. A model is proposed that postulates that P starvation results in (1) an overaccumulation of auxin in the apex of the PR and in young LRs, (2) an overaccumulation of auxin or a change in sensitivity to auxin in the lateral primordia, and (3) a decrease in auxin concentration in the lateral primordia initiation zone of the PR and in old laterals. Measurements of local changes in auxin concentrations induced by low P, either by direct quantification or by biosensor expression pattern (DR5::beta-glucuronidase reporter gene), are in line with these hypotheses. Furthermore, the observation that low P availability mimicked the action of auxin in promoting LR development in the alf3 mutant confirmed that P starvation stimulates primordia emergence through increased accumulation of auxin or change in sensitivity to auxin in the primordia. Both the strong effect of 2,3,5-triiodobenzoic acid and the phenotype of the auxin transport mutants (aux1, eir1) suggest that low P availability modifies local auxin concentrations within the root system through changes in auxin transport rather than auxin synthesis. PMID- 16040661 TI - Effect of temperature on geminivirus-induced RNA silencing in plants. AB - Short-interfering RNAs (siRNAs), the molecular markers of posttranscriptional gene silencing (PTGS), are powerful tools that interfere with gene expression and counter virus infection both in plants and animals. Here, we report the effect of temperature on geminivirus-induced gene silencing by quantifying virus-derived siRNAs and by evaluating their distribution along the virus genome for isolates of five species of cassava geminiviruses in cassava (Manihot esculenta, Crantz) and Nicotiana benthamiana. Cassava geminivirus-induced RNA silencing increased by raising the temperature from 25 degrees C to 30 degrees C, with the appearance of less symptomatic newly developed leaves, irrespective of the nature of the virus. Consequently, nonrecovery-type geminiviruses behaved like recovery-type viruses under high temperature. Next, we evaluated the distribution of virus-derived siRNAs on the respective virus genome at three temperatures (25 degrees C, 25 degrees C-30 degrees C, and 30 degrees C). For recovery-type viruses, siRNAs accumulated at moderately higher levels during virus-induced PTGS at higher temperatures, and there was no change in the distribution of the siRNA population along the virus genome. For nonrecovery-type viruses, siRNAs accumulated at strikingly higher levels than those observed for infections with recovery-type viruses at high temperature. As determined for an RNA virus, temperature influences gene silencing for single-stranded DNA geminiviruses. It is possible that other mechanisms besides gene silencing also control geminivirus accumulation at high temperatures. The findings presented here should be taken into consideration when implementing PTGS-based strategies to control plant virus accumulation. PMID- 16040662 TI - Molecular identification and characterization of the Arabidopsis delta(3,5),delta(2,4)-dienoyl-coenzyme A isomerase, a peroxisomal enzyme participating in the beta-oxidation cycle of unsaturated fatty acids. AB - Degradation of unsaturated fatty acids through the peroxisomal beta-oxidation pathway requires the participation of auxiliary enzymes in addition to the enzymes of the core beta-oxidation cycle. The auxiliary enzyme delta(3,5),delta(2,4)-dienoyl-coenzyme A (CoA) isomerase has been well studied in yeast (Saccharomyces cerevisiae) and mammals, but no plant homolog had been identified and characterized at the biochemical or molecular level. A candidate gene (At5g43280) was identified in Arabidopsis (Arabidopsis thaliana) encoding a protein showing homology to the rat (Rattus norvegicus) delta(3,5),delta(2,4) dienoyl-CoA isomerase, and possessing an enoyl-CoA hydratase/isomerase fingerprint as well as aspartic and glutamic residues shown to be important for catalytic activity of the mammalian enzyme. The protein, named AtDCI1, contains a peroxisome targeting sequence at the C terminus, and fusion of a fluorescent protein to AtDCI1 directed the chimeric protein to the peroxisome in onion (Allium cepa) cells. AtDCI1 expressed in Escherichia coli was shown to have delta(3,5),delta(2,4)-dienoyl-CoA isomerase activity in vitro. Furthermore, using the synthesis of polyhydroxyalkanoate in yeast peroxisomes as an analytical tool to study the beta-oxidation cycle, expression of AtDCI1 was shown to complement the yeast mutant deficient in the delta(3,5),delta(2,4)-dienoyl-CoA isomerase, thus showing that AtDCI1 is also appropriately targeted to the peroxisome in yeast and has delta(3,5),delta(2,4)-dienoyl-CoA isomerase activity in vivo. The AtDCI1 gene is expressed constitutively in several tissues, but expression is particularly induced during seed germination. Proteins showing high homology with AtDCI1 are found in gymnosperms as well as angiosperms belonging to the Monocotyledon or Dicotyledon classes. PMID- 16040663 TI - Virus-induced gene silencing-based functional characterization of genes associated with powdery mildew resistance in barley. AB - We successfully implemented virus-induced gene silencing (VIGS) in barley (Hordeum vulgare) for the functional characterization of genes required for Mla13 mediated resistance toward the biotrophic barley pathogen Blumeria graminis f. sp. hordei. Initially, barley cultivars were screened for their ability to host the barley stripe mosaic virus (BSMV)-VIGS vector by allowing its replication and systemic movement without causing excessive symptoms. Phytoene desaturase silencing leading to photobleaching was used as a phenotypic marker alongside reverse transcription-PCR data to characterize the silencing response at the molecular level. Barley cultivar Clansman, harboring the Mla13 resistance gene, was chosen as the most suitable host for BSMV-VIGS-based functional characterization of Rar1, Sgt1, and Hsp90 in the Mla-mediated resistance toward powdery mildew. BSMV-induced gene silencing of these candidate genes, which are associated in many but not all race-specific pathways, proved to be robust and could be detected at both mRNA and protein levels for up to 21 d postinoculation. Systemic silencing was observed not only in the newly developed leaves from the main stem but also in axillary shoots. By examining fungal development from an incompatible mildew strain carrying the cognate Avr13 gene on plants BSMV silenced for Rar1, Sgt1, and Hsp90, a resistance-breaking phenotype was observed, while plants infected with BSMV control constructs remained resistant. We demonstrate that Hsp90 is a required component for Mla13-mediated race-specific resistance and that BSMV-induced VIGS is a powerful tool to characterize genes involved in pathogen resistance in barley. PMID- 16040665 TI - Functional redundancy of AtFtsH metalloproteases in thylakoid membrane complexes. AB - FtsH is an ATP-dependent metalloprotease found in bacteria, mitochondria, and plastids. Arabidopsis (Arabidopsis thaliana) contains 12 AtFtsH proteins, three in the mitochondrion and nine in the chloroplast. Four of the chloroplast FtsH proteins are encoded by paired members of closely related genes (AtFtsH1 and 5, and AtFtsH2 and 8). We have previously reported that AtFtsH2 and 8 are interchangeable components of AtFtsH complexes in the thylakoid membrane. In this article, we show that the var1 variegation mutant, which is defective in AtFtsH5, has a coordinate reduction in the AtFtsH2 and 8 pair, and that the levels of both pairs are restored to normal in var1 plants that overexpress AtFtsH1. Overexpression of AtFtsH1, but not AtFtsH2/VAR2, normalizes the pattern of var1 variegation, restoring a nonvariegated phenotype. We conclude that AtFtsH proteins within a pair, but not between pairs, are interchangeable and functionally redundant, at least in part. We further propose that the abundance of each pair is matched with that of the other pair, with excess subunits being turned over. The variegation phenotype of var1 (as well as var2, which is defective in AtFtsH2) suggests that a threshold concentration of subunits is required for normal chloroplast function. AtFtsH1, 2, 5, and 8 do not show evidence of tissue or developmental specific expression. Phylogenetic analyses revealed that rice (Oryza sativa) and Arabidopsis share a conserved core of seven FtsH subunit genes, including the AtFtsH1 and 5 and AtFtsH2 and 8 pairs, and that the structure of the present-day gene families can be explained by duplication events in each species following the monocot/dicot divergence. PMID- 16040664 TI - SEC8, a subunit of the putative Arabidopsis exocyst complex, facilitates pollen germination and competitive pollen tube growth. AB - The exocyst, a complex of eight proteins, contributes to the morphogenesis of polarized cells in a broad range of eukaryotes. In these organisms, the exocyst appears to facilitate vesicle docking at the plasma membrane during exocytosis. Although we had identified orthologs for each of the eight exocyst components in Arabidopsis (Arabidopsis thaliana), no function has been demonstrated for any of them in plants. The gene encoding one exocyst component ortholog, AtSEC8, is expressed in pollen and vegetative tissues of Arabidopsis. Genetic studies utilizing an allelic series of six independent T-DNA mutations reveal a role for SEC8 in male gametophyte function. Three T-DNA insertions in SEC8 cause an absolute, male-specific transmission defect that can be complemented by expression of SEC8 from the LAT52 pollen promoter. Microscopic analysis shows no obvious abnormalities in the microgametogenesis of the SEC8 mutants, and the mutant pollen grains appear to respond to the signals that initiate germination. However, in vivo assays indicate that these mutant pollen grains are unable to germinate a pollen tube. The other three T-DNA insertions are associated with a partial transmission defect, such that the mutant allele is transmitted through the pollen at a reduced frequency. The partial transmission defect is only evident when mutant gametophytes must compete with wild-type gametophytes, and arises in part from a reduced pollen tube growth rate. These data support the hypothesis that one function of the putative plant exocyst is to facilitate the initiation and maintenance of the polarized growth of pollen tubes. PMID- 16040667 TI - A randomized clinical trial of activated charcoal for the routine management of oral drug overdose. AB - BACKGROUND: Activated charcoal (AC) is commonly used for the routine management of oral drug overdose. AIM: To determine whether the routine use of activated charcoal has an effect on patient outcomes. DESIGN: Randomized controlled unblinded trial. METHODS: We recruited all adult patients presenting with an oral overdose at The Canberra Hospital, excluding only transfers, late presenters, those who had ingested drugs not adsorbed by activated charcoal or where administration was contraindicated, and very serious ingestions (at the discretion of the admitting physician). Patients were randomized to either activated charcoal or no decontamination. RESULTS: The trial recruited 327 patients over 16 months. Of 411 presentations, four refused consent, 27 were protocol violations and 53 were excluded from the trial. Only seven were excluded due to the severity of their ingestion. The most common substances ingested were benzodiazepines, paracetamol and selective serotonin reuptake inhibitor antidepressants. More than 80% of patients presented within 4 h following ingestion. There were no differences between AC and no decontamination in terms of length of stay (AC 6.75 h, IQR 4-14 vs. controls 5.5 h, IQR 3-12; p=0.11) or secondary outcomes including vomiting, mortality and intensive care admission. DISCUSSION: Routine administration of charcoal following oral overdose did not significantly influence length of stay or other patient outcomes following oral drug overdose. There were few adverse events. This does not exclude a role in patients who present shortly after ingestion of highly lethal drugs. PMID- 16040666 TI - Involvement of ethylene in stress-induced expression of the TLC1.1 retrotransposon from Lycopersicon chilense Dun. AB - The TLC1 family is one of the four families of long terminal repeat (LTR) retrotransposons identified in the genome of Lycopersicon chilense. Here, we show that this family of retroelements is transcriptionally active and its expression is induced in response to diverse stress conditions such as wounding, protoplast preparation, and high salt concentrations. Several stress-associated signaling molecules, including ethylene, methyl jasmonate, salicylic acid, and 2,4 dichlorophenoxyacetic acid, are capable of inducing TLC1 family expression in vivo. A representative of this family, named TLC1.1, was isolated from a genomic library from L. chilense. Transient expression assays in leaf protoplasts and stably transformed tobacco (Nicotiana tabacum) plants demonstrate that the U3 domain of the 5'-LTR region of this element can drive stress-induced transcriptional activation of the beta-glucuronidase reporter gene. Two 57-bp tandem repeated sequences are found in this region, including an 8-bp motif, ATTTCAAA, previously identified as an ethylene-responsive element box in the promoter region of ethylene-induced genes. Expression analysis of wild-type LTR and single and double ethylene-responsive element box mutants fused to the beta glucuronidase gene shows that these elements are required for ethylene-responsive gene expression in protoplasts and transgenic plants. We suggest that ethylene dependent signaling is the main signaling pathway involved in the regulation of the expression of the TLC1.1 element from L. chilense. PMID- 16040668 TI - Primary angiitis of the central nervous system: emerging variants. AB - BACKGROUND: Primary angiitis of the central nervous system (PACNS), a serious disease, has not featured prominently in the spectrum of multi-organ disease seen in vasculitis clinics. AIM: To evaluate the presentation, natural history and features of PACNS variants and compare to those of systemic vasculitides. DESIGN: Retrospective analysis. METHODS: Patients (n=105) presented during 1988-2003 to a tertiary regional vasculitis clinic receiving unselected disease types. Data were collected from a clinical database, patient and laboratory records. RESULTS: The frequency of PACNS presentation rose over the study period, compared with most of the other vasculitides. When PACNS was divided into small- and middle-sized vessel disease (SVD/MVD), their clinical courses differed substantially. SVD PACNS was responsive to immunosuppressive drugs, but relapsed during prolonged periods in all patients on maintenance immunosuppressives, or after withdrawal of treatment, causing recurrent, severe and irreversible CNS injury. MVD PACNS had isolated episodes at presentation, with a paucity of relapses during prolonged follow-up. DISCUSSION: Similarities between SVD PACNS and microscopic polyarteritis suggest the former may represent a limited form of the latter. MVD PACNS has a distinctly more benign relapse pattern than its multisystem counterpart polyarteritis nodosa. Acute-phase serology was useful in designating inflammatory processes at presentation of patients presenting with encephalopathy caused by SVD only, but were unhelpful in defining relapses in this form of PACNS, the definition of which in all cases rested on clinical assessment and MR scanning. Direct cerebral angiography was not diagnostic in any case of SVD PACNS; positive brain biopsy is diagnostically unequivocal, but the total clinical syndrome with imaging may establish a diagnosis with highest probability. In MVD PACNS, angiography with MR scan proved diagnostic. We suggest an algorithm for a rational, minimally invasive approach to investigation. In PACNS, SVD and MVD are important variants, and decisions about therapy should incorporate these distinctions. PMID- 16040669 TI - Coronary stenting and abciximab in primary angioplasty for ST-segment-elevation myocardial infarction. AB - Advances in anti-platelet therapy and improvement of stent deployment techniques have improved the safety and efficacy of stenting in the setting of ST-segment elevation myocardial infarction (STEMI). However, in randomized trials, routine coronary stenting does not reduce mortality and re-infarction, compared to balloon angioplasty. Further, the benefits in target vessel revascularization seem to be reduced when applied to unselected patients with STEMI. Direct stenting represents an attractive strategy with potential benefits in terms of myocardial perfusion. Future large randomized trials are needed to evaluate whether this strategy has a significant impact on outcome, and to provide a cost benefit analysis of the unrestricted use of drug-eluting stents in this high-risk subset of patients. The additional use of abciximab reduces mortality in primary angioplasty. Since the feasibility of long-distance transportation has been shown in several randomized trials, early pharmacological pre-treatment may confer further advantages by early recanalization and shorter ischaemic time, particularly in high-risk patients. Further randomized trials are needed to clarify the potential benefits from early abciximab administration and the potential role of small molecules in primary angioplasty for STEMI. PMID- 16040670 TI - Using goal attainment scaling to improve the quality of long-term care: a group randomized trial. AB - OBJECTIVE: To assess the effect of a 6-month interdisciplinary customized intervention based on goal attainment scaling and aimed at improving the quality of care provided by long-term care facilities to frail older adults. DESIGN: A pair-matched group-randomized trial with quality of care measurements taken before and after the intervention and 6 months later. SETTING: Forty unregulated small-to-medium sized long-term care facilities delivering inadequate care to at least one resident. Facilities were matched on baseline quality of care, health district, and size. One member of each pair was then randomly allocated to the intervention, the other acting as a control. STUDY PARTICIPANTS: The trial involved 201 frail older adults randomly selected from the 40 participating facilities. INTERVENTION: The intervention was tailored to the main quality problems identified at baseline in the facility. The first task of the intervention team was to set weighted quality improvement goals with the facility manager, which were then recorded on a goal attainment follow-up guide. Subsequent monthly on-site visits, interspersed with frequent telephone calls, were meant to assist the manager and staff to implement permanent changes in the areas of care targeted for improvement. Main outcome measure. Quality of care measured with the Quality of Care (QUALCARE) Scale, a multidimensional instrument that uses a 5-point scale to assess six subdimensions of care: environmental, physical, medical management, psychosocial, human rights, and financial. A score greater than 2 is considered indicative of inadequate care. RESULTS: The intervention effect on the overall quality of care was neither clinically nor statistically significant. Change from baseline to the end of the intervention averaged .21 and .22 in the experimental and control conditions, respectively (P = 0.86). CONCLUSION: Attainment of preset quality improvement objectives did not translate into detectable improvements in the care provided to residents. PMID- 16040671 TI - Misusing the Nazi analogy. PMID- 16040672 TI - Human space flight. NASA may cut shuttle flights and reduce science on station. PMID- 16040673 TI - Immunology. New virtual center aims to speed AIDS vaccine progress. PMID- 16040675 TI - Italian science. Carlo Rubbia dismissed from energy agency. PMID- 16040674 TI - U.K. science policy. Parliamentary gadfly loses his post. PMID- 16040677 TI - UNESCO. U.S. rules could muffle scientific voices. PMID- 16040676 TI - Science and law. Flawed statistics in murder trial may cost expert his medical license. PMID- 16040678 TI - Antiterrorism. Defense rules would pinch foreign-born scientists. PMID- 16040679 TI - National Institutes of Health. Bill could restructure agency and strengthen director's hand. PMID- 16040680 TI - Ecology. Global analyses reveal mammals facing risk of extinction. PMID- 16040681 TI - Conflict of interest. Forty-four researchers broke NIH consulting rules. PMID- 16040682 TI - Research funding. France hatches 67 California wannabes. PMID- 16040683 TI - Solar energy. Is it time to shoot for the sun? PMID- 16040684 TI - Solar energy. Solar report sets the agenda. PMID- 16040685 TI - Reproductive biology. A powerful first KiSS-1. PMID- 16040686 TI - Astronomy. Europe joins forces in push for monster scope project. PMID- 16040687 TI - The origins of Olmec civilization. PMID- 16040688 TI - "Intelligent" design versus evolution. PMID- 16040689 TI - Issues in Indian science. PMID- 16040690 TI - Comment on "Nervy links protein kinase A to plexin-mediated semaphorin repulsion". PMID- 16040692 TI - Ecology. North Atlantic right whales in crisis. PMID- 16040693 TI - Sustainability. Millennium assessment of human behavior. PMID- 16040694 TI - Astronomy. Mapping the large-scale structure of the universe. PMID- 16040695 TI - Physics. Fingerprinting spin qubits. PMID- 16040696 TI - Ecology. Population dynamics: growing to extremes. PMID- 16040697 TI - Neuroscience. Similar is different in hippocampal networks. PMID- 16040698 TI - Global consequences of land use. AB - Land use has generally been considered a local environmental issue, but it is becoming a force of global importance. Worldwide changes to forests, farmlands, waterways, and air are being driven by the need to provide food, fiber, water, and shelter to more than six billion people. Global croplands, pastures, plantations, and urban areas have expanded in recent decades, accompanied by large increases in energy, water, and fertilizer consumption, along with considerable losses of biodiversity. Such changes in land use have enabled humans to appropriate an increasing share of the planet's resources, but they also potentially undermine the capacity of ecosystems to sustain food production, maintain freshwater and forest resources, regulate climate and air quality, and ameliorate infectious diseases. We face the challenge of managing trade-offs between immediate human needs and maintaining the capacity of the biosphere to provide goods and services in the long term. PMID- 16040699 TI - Web-spinning caterpillar stalks snails. AB - Moths and butterflies compose one of the most diverse insect orders, but they are overwhelmingly herbivorous. Less than 0.2% are specialized predators, indicating that lepidopteran feeding habits are highly constrained. We report a Hawaiian caterpillar that specializes on snails, a unique food source requiring an unusual feeding strategy. The caterpillar uses silk to restrain live prey. All caterpillars have silk glands, but none are known to use silk in this spiderlike fashion. Considering the canalization of caterpillar diets, evolution to attack and feed on snails is an anomaly. Hawaii s isolation and consequently disharmonic biota likely promote evolutionary experiments that occur nowhere else. PMID- 16040700 TI - Fermionic bell-state analyzer for spin qubits. AB - We propose a protocol and physical implementation for partial Bell-state measurements of Fermionic qubits, allowing for deterministic quantum computing in solid-state systems without the need for two-qubit gates. Our scheme consists of two spin qubits in a double quantum dot where the two dots have different Zeeman splittings and resonant tunneling between the dots is only allowed when the spins are antiparallel. This converts spin parity into charge information by means of a projective measurement and can be implemented with established technologies. This measurement-based qubit scheme greatly simplifies the experimental realization of scalable quantum computers in electronic nanostructures. PMID- 16040701 TI - Creating, varying, and growing single-site molecular contacts. AB - The known range of chemisorption bonds forms the toolbox for the design of electrical contacts in molecular electronics devices. Double-bond contacts to technologically relevant materials would be attractive for a number of reasons. They are truly single-site, bonding to a single surface atom. They obviate the need for a thiol linkage, and they may be amenable to further modification through olefin-metathesis methodologies. We report olefin-metathesis methods for establishing, varying, and growing thermally stable double-bond contacts to molybdenum carbide, a conducting material. PMID- 16040702 TI - Formation of catalytic metal-molecule contacts. AB - We describe a new strategy for the in situ growth of molecular wires predicated on the synthesis of a trifunctional "primed" contact formed from metal-carbon multiple bonds. The ruthenium-carbon pi bond provides structural stability to the molecular linkages under ambient conditions, and density functional calculations indicate the formation of an efficient conduit for charge carriers to pass between the metal and the molecule. Moreover, the metal-carbon pi bond provides a chemically reactive site from which a conjugated molecular wire can be grown in situ through an olefin metathesis reaction. PMID- 16040703 TI - Martian surface paleotemperatures from thermochronology of meteorites. AB - The temporal evolution of past martian surface temperatures is poorly known. We used thermochronology and published noble gas and petrographic data to constrain the temperature histories of the nakhlites and martian meteorite ALH84001. We found that the nakhlites have not been heated to more than 350 degrees C since they formed. Our calculations also suggest that for most of the past 4 billion years, ambient near-surface temperatures on Mars are unlikely to have been much higher than the present cold (<0 degrees C) state. PMID- 16040704 TI - Global mammal conservation: what must we manage? AB - We present a global conservation analysis for an entire "flagship" taxon, land mammals. A combination of rarity, anthropogenic impacts, and political endemism has put about a quarter of terrestrial mammal species, and a larger fraction of their populations, at risk of extinction. A new global database and complementarity analysis for selecting priority areas for conservation shows that approximately 11% of Earth's land surface should be managed for conservation to preserve at least 10% of terrestrial mammal geographic ranges. Different approaches, from protection (or establishment) of reserves to countryside biogeographic enhancement of human-dominated landscapes, will be required to approach this minimal goal. PMID- 16040705 TI - On the regulation of populations of mammals, birds, fish, and insects. AB - A key unresolved question in population ecology concerns the relationship between a population's size and its growth rate. We estimated this relationship for 1780 time series of mammals, birds, fish, and insects. We found that rates of population growth are high at low population densities but, contrary to previous predictions, decline rapidly with increasing population size and then flatten out, for all four taxa. This produces a strongly concave relationship between a population's growth rate and its size. These findings have fundamental implications for our understanding of animals' lives, suggesting in particular that many animals in these taxa will be found living at densities above the carrying capacity of their environments. PMID- 16040706 TI - Host suppression and stability in a parasitoid-host system: experimental demonstration. AB - We elucidate the mechanisms causing stability and severe resource suppression in a consumer-resource system. The consumer, the parasitoid Aphytis, rapidly controlled an experimentally induced outbreak of the resource, California red scale, an agricultural pest, and imposed a low, stable pest equilibrium. The results are well predicted by a mechanistic, independently parameterized model. The key mechanisms are widespread in nature: an invulnerable adult stage in the resource population and rapid consumer development. Stability in this biologically nondiverse agricultural system is a property of the local interaction between these two species, not of spatial processes or of the larger ecological community. PMID- 16040707 TI - Dynamics of mammalian chromosome evolution inferred from multispecies comparative maps. AB - The genome organizations of eight phylogenetically distinct species from five mammalian orders were compared in order to address fundamental questions relating to mammalian chromosomal evolution. Rates of chromosome evolution within mammalian orders were found to increase since the Cretaceous-Tertiary boundary. Nearly 20% of chromosome breakpoint regions were reused during mammalian evolution; these reuse sites are also enriched for centromeres. Analysis of gene content in and around evolutionary breakpoint regions revealed increased gene density relative to the genome-wide average. We found that segmental duplications populate the majority of primate-specific breakpoints and often flank inverted chromosome segments, implicating their role in chromosomal rearrangement. PMID- 16040708 TI - Extreme reversed sexual dichromatism in a bird without sex role reversal. AB - Brilliant plumage is typical of male birds, reflecting differential enhancement of male traits when females are the limiting sex. Brighter females are thought to evolve exclusively in response to sex role reversal. The striking reversed plumage dichromatism of Eclectus roratus parrots does not fit this pattern. We quantify plumage color in this species and show that very different selection pressures are acting on males and females. Male plumage reflects a compromise between the conflicting requirements for camouflage from predators while foraging and conspicuousness during display. Females are liberated from the need for camouflage but compete for rare nest hollows. PMID- 16040709 TI - Independent codes for spatial and episodic memory in hippocampal neuronal ensembles. AB - Hippocampal neurons were recorded under conditions in which the recording chamber was varied but its location remained unchanged versus conditions in which an identical chamber was encountered in different places. Two forms of neuronal pattern separation occurred. In the variable cue-constant place condition, the firing rates of active cells varied, often over more than an order of magnitude, whereas the location of firing remained constant. In the variable place-constant cue condition, both location and rates changed, so that population vectors for a given location in the chamber were statistically independent. These independent encoding schemes may enable simultaneous representation of spatial and episodic memory information. PMID- 16040710 TI - Plant circadian clocks increase photosynthesis, growth, survival, and competitive advantage. AB - Circadian clocks are believed to confer an advantage to plants, but the nature of that advantage has been unknown. We show that a substantial photosynthetic advantage is conferred by correct matching of the circadian clock period with that of the external light-dark cycle. In wild type and in long- and short circadian period mutants of Arabidopsis thaliana, plants with a clock period matched to the environment contain more chlorophyll, fix more carbon, grow faster, and survive better than plants with circadian periods differing from their environment. This explains why plants gain advantage from circadian control. PMID- 16040711 TI - Ischemic preconditioning enhances scavenging activity of reactive oxygen species and diminishes transmural difference of infarct size. AB - Reactive oxygen species (ROS) enhance myocardial ischemia-reperfusion (I/R) injury. Ischemic preconditioning (PC) provides potent cardioprotective effects in I/R. However, it has not been elucidated whether PC diminishes ROS stress in I/R and whether PC protects the myocardium from ROS stress transmurally and homogeneously. Isolated rabbit hearts perfused with Krebs-Henseleit buffer underwent 30 min of ischemia and 60 min of reperfusion. Hemodynamic changes and myocardial damage extent were analyzed in four groups. The control group underwent I/R alone. The H2O2 group underwent I/R with H2O2 infusion (50 microM) in the first minute of reperfusion to enhance oxidative stress. The PC and H2O2+PC groups underwent 5 min of PC before control and H2O2 protocols, respectively. Extracted myocardial DNA was analyzed for 8-hydroxydeoxyguanosine (8-OHdG), an indicator of oxidative DNA damage, with the use of the HPLC electrochemical detection method. Glutathione peroxidase (GPX) activity and the reduced form of GSH were measured by spectrophotometric assays. The myocardial infarct size was significantly reduced in the PC group (19 +/- 2%) compared with the control group (37 +/- 4%; P < 0.05), particularly in the subendocardium. H2O2 transmurally increased the infarct size by 59 +/- 4% (P < 0.05), which was significantly diminished in the H2O2+PC group (31 +/- 4%; P < 0.01). The GSH levels, but not GPX activity, were well preserved transmurally in protocols with PC. The 8-OHdG levels were significantly decreased in PC and were significantly enhanced in H2O2 (P < 0.01). These changes in oxidative DNA damage were effectively diminished by PC. In conclusion, PC enhanced the scavenging activity of GSH against ROS transmurally, reduced myocardial damage, particularly in the subendocardium, and diminished the transmural difference in myocardial infarct size. PMID- 16040712 TI - Acetylcholine and sodium nitroprusside cause long-term inhibition of EDCF mediated contractions. AB - Preliminary studies suggested that previous exposure to acetylcholine (ACh) exerts a delayed inhibition of subsequent contractions mediated by endothelium derived contracting factor (EDCF). To confirm this long-term inhibitory effect of ACh and to determine whether nitric oxide (NO) mediates the phenomenon, we suspended rings of spontaneously hypertensive rat (SHR) aortas in organ chambers for the recording of isometric force. The rings were incubated in the absence or presence of Nomega-nitro-L-arginine methyl ester (L-NAME; inhibitor of NO synthases) or 1H-[1,2,4]oxadiazolo[4,3-alpha]quinoxalin-1-one (ODQ; inhibitor of soluble guanylyl cyclase) before exposure to increasing concentrations of ACh or sodium nitroprusside (SNP) during contractions to phenylephrine. Thereafter, EDCF mediated contractions to ACh or the calcium ionophore A-23187 were elicited. If the rings were preexposed to ACh or SNP, the subsequent ACh-induced EDCF-mediated contractions were reduced compared with those obtained in rings of the same arteries not previously exposed to either agent. ODQ did not affect the inhibition caused by preexposure to ACh but significantly reduced that caused by preexposure to SNP. Previous exposure to SNP reduced, whereas previous exposure to ACh did not affect, endothelium-dependent contractions to A-23187. Previous exposure to either ACh or SNP did not affect the contractions to the thromboxane mimetic U-46619. Thus ACh and SNP exert delayed inhibition of EDCF-mediated contractions via distinct pathways. The effect of ACh is NO independent and upstream of the increase in calcium concentration that triggers the release of EDCF. The effect of SNP is downstream of the calcium rise and is mainly NO dependent. PMID- 16040713 TI - Role of F-actin organization in p38 MAP kinase-mediated apoptosis and necrosis in neonatal rat cardiomyocytes subjected to simulated ischemia and reoxygenation. AB - Activation of p38 mitogen-activated protein (MAP) kinase (MAPK) has been implicated in the mechanism of cardiomyocyte (CMC) protection and injury. The p38 MAPK controversy may be related to differential effects of this kinase on apoptosis and necrosis. We have hypothesized that p38 MAPK-mediated F-actin reorganization promotes apoptotic cell death, whereas it protects from osmotic stress-induced necrotic cell death. Cultured neonatal rat CMCs were subjected to 2 h of simulated ischemia followed by reoxygenation. p38 MAPK activity measured by phosphorylation of MAP kinase-activated protein (MAPKAP) kinase 2 was increased during simulated ischemia and reoxygenation. This was associated with translocation of heat shock protein 27 (HSP27) from the cytosolic to the cytoskeletal fraction and F-actin reorganization. Cytochrome c release from mitochondria, caspase-3 activation, and DNA fragmentation were increased during reoxygenation. Robust lactate dehydrogenase (LDH) release was observed under hyposmotic (140 mosM) reoxygenation. The p38 MAPK inhibitor SB-203580 abrogated activation of p38 MAPK, translocation of HSP27, and F-actin reorganization and prevented cytochrome c release, caspase-3 activation, and DNA fragmentation. Conversely, SB-203580 enhanced LDH release during hyposmotic reoxygenation. The F actin disrupting agent cytochalasin D inhibited F-actin reorganization and prevented cytochrome c release, caspase-3 activation, and DNA fragmentation, whereas it enhanced LDH release during hyposmotic reoxygenation. When CMCs were incubated under the isosmotic condition for the first 15 min of reoxygenation, SB 203580 and cytochalasin D increased ATP content of CMCs and prevented LDH release after the conversion to the hyposmotic condition. These results suggest that F actin reorganization mediated by activation of p38 MAPK plays a differential role in apoptosis and protection against osmotic stress-induced necrosis during reoxygenation in neonatal rat CMCs; however, the sarcolemmal fragility caused by p38 MAPK inhibition can be reversed during temporary blockade of physical stress during reoxygenation. PMID- 16040715 TI - Hyaluronidase treatment of coronary glycocalyx increases reactive hyperemia but not adenosine hyperemia in dog hearts. AB - Because adenosine is commonly used for inducing maximal coronary hyperemia in the clinic, it is imperative that adenosine-induced hyperemia (AH) resembles coronary hyperemia that can be attained by endogenous stimuli. In the present study we hypothesized that coronary reactive hyperemia (RH) is limited compared with AH due to the presence of the glycocalyx and that the AH response is therefore unable to detect glycocalyx modifications. In anesthetized open-chest dogs, blood flow and pressure were measured in the left circumflex artery. RH after 15-s occlusion was compared with an intracoronary infusion of adenosine (650 microg; AH) during control conditions and after intracoronary treatment of the glycocalyx with hyaluronidase (20.000 U, 2 x 20 min; n = 6) or heat-inactivated hyaluronidase (n = 5). During control, coronary conductance during RH was 1.49 +/ 0.15 ml.mmHg(-1).min(-1) and 76 +/- 7% of coronary conductance during AH (P < 0.05). After hyaluronidase, RH conductance increased (P < 0.01) by 43 +/- 13% and became 93 +/- 4% of AH conductance (P = NS). Heat-inactivated hyaluronidase had no effect on RH and AH conductance. Our results demonstrate that adenosine induced coronary hyperemia profoundly exceeds RH and that the difference is virtually abolished on selective removal of the glycocalyx. It is concluded that, compared with RH, adenosine-induced coronary hyperemia is not affected by modification of the glycocalyx. This glycocalyx insensitivity should be taken into account when using adenosine-induced coronary hyperemia as a marker for vasodilating capacity to an ischemic stimulus. PMID- 16040714 TI - Effect of vitamin C and L-NMMA on the inotropic response to dobutamine in patients with heart failure. AB - The positive effect of vitamin C on left ventricular (LV) inotropic responses to dobutamine, observed in patients with preserved LV function, is lost in heart failure (HF). We tested the hypothesis that in HF, endogenous nitric oxide (NO) opposes the positive effect of vitamin C on adrenergically stimulated contractility by examining the effects of vitamin C on dobutamine responses during NO synthase inhibition. In 11 HF patients, a micromanometer-tipped catheter was inserted into the LV and an infusion catheter was positioned in the left main coronary artery. The peak positive rate of change of LV pressure (LV +dP/dt) was measured in response to intravenous dobutamine (Dob-1). After recontrol, intracoronary N(G)-monomethyl-L-arginine (l-NMMA) was infused before reinfusion of dobutamine (L-NMMA + Dob-2). Finally, intracoronary vitamin C was infused in addition to intracoronary L-NMMA and dobutamine (L-NMMA + Dob-2 + vitamin C). Intracoronary L-NMMA alone had no effect on LV +dP/dt. After a stable inotropic response to intracoronary L-NMMA and dobutamine was established, the addition of intracoronary vitamin C resulted in a modest but significant increase in LV +dP/dt. The change in LV +dP/dt in response to dobutamine alone was 25 +/- 5%, with intracoronary L-NMMA, 27 +/- 6%, and with intracoronary L-NMMA plus vitamin C, 37 +/- 5% (P < 0.05 vs. Dob-1 and L-NMMA + Dob-2). These findings demonstrate that an interaction between endogenous NO and redox environment exists and exerts some influence on stimulated contractility in HF. PMID- 16040716 TI - Nitric oxide modulates oxygen consumption by arteriolar walls in rat skeletal muscle. AB - To study the role of nitric oxide (NO) in regulating oxygen consumption by vessel walls, the oxygen consumption rate of arteriolar walls in rat cremaster muscle was measured in vivo during flow-induced vasodilation and after inhibiting NO synthesis. The oxygen consumption rate of arteriolar walls was calculated based on the intra- and perivascular PO2 values measured by phosphorescence quenching laser microscopy. The perivascular PO2 value of the arterioles during vasodilation was significantly higher than under control conditions, although the intravascular PO2 values under both conditions were approximately the same. Inhibition of NO synthesis, on the other hand, caused a significant increase in arterial blood pressure and a significant decrease in arteriolar diameter. Inhibition of NO synthesis also caused a significant decrease in both the intra- and perivascular PO2 values of the arterioles. Inhibition of NO synthesis increased the oxygen consumption rate of the vessel walls by 42%, whereas enhancement of flow-induced NO release decreased it by 34%. These results suggest that NO plays an important role not only as a regulator of peripheral vascular tone but also as a modulator of tissue oxygenation by reducing oxygen consumption by vessel walls. In addition, enhancement of NO release during exercise may facilitate efficient oxygen supply to the surrounding high metabolic tissue. PMID- 16040717 TI - Cardioprotection induced by cardiac-specific overexpression of fibroblast growth factor-2 is mediated by the MAPK cascade. AB - Our laboratory showed previously that cardiac-specific overexpression of FGF-2 [FGF-2 transgenic (Tg)] results in increased recovery of contractile function and decreased infarct size after ischemia-reperfusion injury. MAPK signaling is downstream of FGF-2 and has been implicated in other models of cardioprotection. Treatment of FGF-2 Tg and wild-type hearts with U-0126, a MEK-ERK pathway inhibitor, significantly reduced recovery of contractile function after global low-flow ischemia-reperfusion injury in FGF-2 Tg (86 +/- 2% vehicle vs. 66 +/- 4% U-0126; P < 0.05) but not wild-type (61 +/- 7% vehicle vs. 67 +/- 7% U-0126) hearts. Similarly, MEK-ERK inhibition significantly increased myocardial infarct size in FGF-2 Tg (12 +/- 3% vehicle vs. 31 +/- 2% U-0126; P < 0.05) but not wild type (30 +/- 4% vehicle vs. 36 +/- 7% U-0126) hearts. In contrast, treatment of FGF-2 Tg and wild-type hearts with SB-203580, a p38 inhibitor, did not abrogate FGF-2-induced cardioprotection from postischemic contractile dysfunction. Instead, inhibition of p38 resulted in decreased infarct size in wild-type hearts (30 +/- 4% vehicle vs. 11 +/- 2% SB-203580; P < 0.05) but did not alter infarct size in FGF-2 Tg hearts (12 +/- 3% vehicle vs. 14 +/- 1% SB-203580). Western blot analysis of ERK and p38 activation revealed signaling alterations in FGF-2 Tg and wild-type hearts during early ischemia or reperfusion injury. In addition, MEK independent ERK inhibition by p38 was observed during early ischemic injury. Together these data suggest that activation of ERK and inhibition of p38 by FGF-2 is cardioprotective during ischemia-reperfusion injury. PMID- 16040718 TI - Hybrid duplex: a novel method to study the contractile function of heterogeneous myocardium. AB - In an earlier study, we experimentally mimicked the effects of mechanical interaction between different regions of the ventricular wall by allowing pairs of independently maintained cardiac muscle fibers to interact mechanically in series or in parallel. This simple physiological model of heterogeneous myocardium, which has been termed "duplex," has provided new insight into basic effects of cardiac electromechanical heterogeneity. Here, we present a novel "hybrid duplex," where one of the elements is an isolated cardiac muscle and the other a "virtual cardiac muscle." The virtual muscle is represented by a computational model of cardiomyocyte electromechanical activity. We present in detail the computer-based digital control system that governs the mechanical interaction between virtual and biological muscle, the software used for data analysis, and working implementations of the model. Advantages of the hybrid duplex method are discussed, and experimental recordings are presented for illustration and as proof of the principle. PMID- 16040719 TI - Microvascular blood viscosity in vivo and the endothelial surface layer. AB - The apparent viscosity of blood in glass tubes declines with decreasing diameter (Fahraeus-Lindqvist effect) and exhibits a distinctive minimum at 6-7 microm. However, flow resistance in vivo in small vessels is substantially higher than predicted by in vitro viscosity data. The presence of a thick endothelial surface layer (ESL) has been proposed as the primary cause for this discrepancy. Here, a physical model is proposed for microvascular flow resistance as a function of vessel diameter and hematocrit in vivo; it combines in vitro blood viscosity with effects of a diameter-dependent ESL. The model was developed on the basis of flow distributions observed in three microvascular networks in the rat mesentery with 392, 546, and 383 vessel segments, for which vessel diameters, network architecture, flow velocity, and hematocrit were determined by intravital microscopy. A previously described hemodynamic simulation was used to predict the distributions of flow and hematocrit from the assumed model for effective blood viscosity. The dependence of ESL thickness on vessel diameter was estimated by minimizing deviations of predicted values for velocities, flow directions, and hematocrits from measured data. Optimal results were obtained with a layer thickness of approximately 0.8-1 microm for 10- to 40-microm-diameter vessels and declined strongly for smaller diameters, with an additional hematocrit-dependent impact on flow resistance exhibiting a maximum for approximately 10-microm diameter vessels. These results show that flow resistance in vivo can be explained by in vitro blood viscosity and the presence of an ESL and indicate the rheologically effective thickness of the ESL in microvessels. PMID- 16040720 TI - Regulation of cardiac volume-sensitive chloride channel by focal adhesion kinase and Src kinase. AB - The volume-sensitive chloride current (ICl,swell) is found in the mammalian myocardium and is activated by osmotic swelling. The goal of this study was to examine the importance of the tyrosine kinases focal adhesion kinase (FAK) and Src kinase in cardiac ICl,swell regulation. Neonatal rat ventricular myocytes were cultured on collagen membranes and infected with adenovirus expressing beta galactosidase (AdLacZ), FAK, or FAK-related nonkinase. FAK-related nonkinase (FRNK) is an endogenous cardiac protein, which functions as an inhibitor of FAK. Whole cell patch-clamp recordings demonstrated that osmotic swelling was associated with the activation of an outward rectifying current in uninfected and AdLacZ-infected cells. Consistent with the properties of ICl,swell, this current displayed a reversal potential close to the equilibrium potential for Cl-; was inhibited by the Cl- channel blockers 4,4'-dinitrostilbene-2,2'-disulfonic acid, 5-nitro-2-(3-phenylpropylamino)-benzoic acid, and tamoxifen; and was eliminated in hypertonic solution. In addition to activating ICl,swell, hypotonic swelling enhanced the tyrosine phosphorylation of multiple cardiac proteins including those in the range of 68-70 and 120-130 kDa. Pretreatment of the cells with the drug 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine, an inhibitor of FAK and Src, diminished swelling-induced phosphorylation of these proteins but paradoxically increased ICl,swell. Furthermore, overexpression of FRNK but not FAK caused a twofold augmentation in I(Cl,swell) and increased the rate of current activation. Thus the tyrosine kinases FAK and Src contribute to the regulation of ICl,swell. PMID- 16040721 TI - Upregulation of adrenomedullin and its receptor components during cardiomyocyte hypertrophy induced by chronic inhibition of nitric oxide synthesis in rats. AB - Adrenomedullin may provide a compensatory mechanism to attenuate left ventricular hypertrophy (LVH). Nitric oxide synthase inhibition, induced by chronic administration of N(omega)-nitro-L-arginine methyl ester (L-NAME) to rats, induces cardiac hypertrophy in some, but not all cases; there are few reports of direct assessment of cardiomyocyte parameters. The objective was to characterize hypertrophic parameters in left (LV) and right ventricular (RV) cardiomyocytes after administration of L-NAME to rats for 8 wk and to determine whether adrenomedullin and its receptor components were upregulated. After treatment with L-NAME (20 and 50 mg x kg(-1) x day(-1)), compared with nontreated animals, 1) systolic blood pressure increased (by 34.2 and 104.9 mmHg), 2) heart weight-to body wt ratio increased 24.1% at the higher dose (P < 0.05), 3) cardiomyocyte protein mass increased (P = NS), 4) cardiomyocyte protein synthesis ([14C]phenylalanine incorporation) increased (P < 0.05), 5) expression of skeletal alpha-actin, atrial natriuretic peptide, brain natriuretic peptide, and ET-1 mRNAs was enhanced (P < 0.05) in LV but not RV cardiomyocytes at 20 and 50 mg x kg(-1) x day(-1), respectively, and 6) expression of adrenomedullin, receptor activity-modifying protein 3 (RAMP3), and RAMP2 (but not calcitonin receptor-like receptor and RAMP1) mRNAs was increased by L-NAME (20 mg x kg(-1) x day(-1)) in LV. In conclusion, L-NAME enhanced protein synthesis in both LV and RV cardiomyocytes but elicited a hypertrophic phenotype accompanied by altered expression of the counterregulatory peptide adrenomedullin and receptor components (RAMP2, RAMP3) in LV only, indicating that the former is due to impaired nitric oxide synthesis, whereas the phenotypic changes are due to pressure overload. PMID- 16040722 TI - Differential cardioprotective/cardiotoxic effects mediated by beta-adrenergic receptor subtypes. AB - Recent data suggest that beta-adrenergic receptor subtypes couple differentially to signaling pathways regulating cardiac function vs. cardiac remodeling. To dissect the roles of beta1- vs. beta2-receptors in the pathogenesis of cardiomyopathy, doxorubicin was administered to beta1, beta2, and beta1/beta2 knockout (-/-) and wild-type mice. Expression and activation of MAPKs were measured. Wild-type and beta1-/- mice showed no acute cardiovascular effects, whereas beta2-/- mice all died within 30 min. The additional deletion of the beta1-receptor (beta1/beta2-/-) totally rescued this toxicity. beta2-/- mice developed decreased contractile function, hypotension, QTc prolongation, and ST segment changes and a 20-fold increase in p38 MAPK activity not seen in the other genotypes. The MAPK inhibitor SB-203580 rescued beta2-/- mice from this acute toxicity. The enhanced toxicity in beta2-/- mice was also recapitulated in wild type mice with the beta2-selective antagonist ICI-118,551, although the rescue effect of the beta1-deletion was not recapitulated using the beta1-selective antagonist metoprolol or the nonselective beta-antagonist propranolol. These data suggest that beta2-adrenergic receptors play a cardioprotective role in the pathogenesis of cardiomyopathy, whereas beta1-adrenergic receptors mediate at least some of the acute cardiotoxicity of anthracyclines. Differential activation of MAPK isoforms, previously shown in vitro to regulate beta-agonist as well as doxorubicin cardiotoxicity, appears to play a role in mediating the differential effects of these beta-adrenergic receptor subtypes in vivo. PMID- 16040723 TI - Epithelial-mesenchymal transformation during craniofacial development. AB - Epithelial to mesenchymal phenotype transition is a common phenomenon during embryonic development, wound healing, and tumor metastasis. This transition involves cellular changes in cytoskeleton architecture and protein expression. Specifically, this highly regulated biological event plays several important roles during craniofacial development. This review focuses on the regulation of epithelial-mesenchymal transformation (EMT) during neural crest cell migration, and fusion of the secondary palate and the upper lip. PMID- 16040724 TI - The adaptive remodeling of condylar cartilage---a transition from chondrogenesis to osteogenesis. AB - Mandibular condylar cartilage is categorized as articular cartilage but markedly distinguishes itself in many biological aspects, such as its embryonic origin, ontogenetic development, post-natal growth mode, and histological structures. The most marked uniqueness of condylar cartilage lies in its capability of adaptive remodeling in response to external stimuli during or after natural growth. The adaptation of condylar cartilage to mandibular forward positioning constitutes the fundamental rationale for orthodontic functional therapy, which partially contributes to the correction of jaw discrepancies by achieving mandibular growth modification. The adaptive remodeling of condylar cartilage proceeds with the biomolecular pathway initiating from chondrogenesis and finalizing with osteogenesis. During condylar adaptation, chondrogenesis is activated when the external stimuli, e.g., condylar repositioning, generate the differentiation of mesenchymal cells in the articular layer of cartilage into chondrocytes, which proliferate and then progressively mature into hypertrophic cells. The expression of regulatory growth factors, which govern and control phenotypic conversions of chondrocytes during chondrogenesis, increases during adaptive remodeling to enhance the transition from chondrogenesis into osteogenesis, a process in which hypertrophic chondrocytes and matrices degrade and are replaced by bone. The transition is also sustained by increased neovascularization, which brings in osteoblasts that finally result in new bone formation beneath the degraded cartilage. PMID- 16040726 TI - Smokeless tobacco and severe active periodontal disease, NHANES III. AB - Whereas smoking is a major risk factor for periodontal disease, the role of smokeless tobacco is unclear. The purpose of this US population-based study of 12,932 adults participating in the Third National Health and Nutrition Examination Survey was to evaluate the association between smokeless tobacco use and severe active periodontal disease. Univariable and multivariable logistic regression modeling quantified the associations between tobacco use and severe active periodontal disease. All adults and never-smokers who currently used smokeless tobacco were twice as likely to have severe active periodontal disease at any site [respective odds ratios (OR(Adj)) and 95% confidence intervals: OR(Adj) = 2.1; 1.2-3.7 and OR(Adj) = 2.1; 1.0-4.4] or restricted to any interproximal site [respective OR(Adj) = 2.1; 1.0-4.2 and OR(Adj) = 2.3; 0.9 6.3], simultaneously adjusted for smoking, age, race, gender, diabetes, and having a dental visit in the past year. These results indicate that smokeless tobacco may also be an important risk factor for severe active periodontal disease. PMID- 16040725 TI - Fusobacterium nucleatum apoptosis-inducing outer membrane protein. AB - The periodontal pathogen Fusobacterium nucleatum induces apoptosis in lymphocytes. We previously identified the autotransporter protein Fap2 in F. nucleatum strain PK1594 that induced apoptosis in lymphocytes when expressed in Escherichia coli. In this study, we identified protein homologs of Fap2 in the transformable F. nucleatum strain ATCC 23726, to determine their role in the induction of apoptosis in lymphocytes. We used a new gene-inactivation vector conferring thiamphenicol resistance (pHS31) to construct a mutant deficient in one of the homologs, aim1. Transcriptional analyses demonstrated disruption of aim1 expression, and phenotypic analyses revealed a 41% decrease in the ability of the mutant to induce apoptosis in Jurkat cells, as compared with the parental strain. These studies demonstrate, in the native host cell background, the contribution of aim1 to F. nucleatum induction of apoptosis and, to the best of our knowledge, represent the first report of a genetically defined and phenotypically characterized mutation in F. nucleatum. PMID- 16040727 TI - Tuftelin, mutans streptococci, and dental caries susceptibility. AB - The purpose of this study was to identify genetic factors that contribute to dental caries susceptibility, either alone or in combination with environmental factors. Dental examinations were performed and buccal swab samples collected from 3- to 5-year-old children with at least 4 surfaces of decay, or with no evidence of decay. SNP assays for each of 6 candidate genes were performed for all cases and controls. Chi-square analysis and regression analysis were used for the evaluation of individual gene effects, environmental effects, and gene environment interactions. There were no significant associations between single candidate genes and caries susceptibility. Levels of S. mutans were positively and Lactobacilli were negatively associated with caries. Regression analysis revealed a significant interaction between tuftelin and S. mutans, with 26.8% of the variation in dmfs explained by the interaction. Future research will focus on the identification of these additional factors and the development of functional assays so that these interactions can be better understood. PMID- 16040728 TI - New coronal caries in older adults: implications for prevention. AB - To characterize the extent and severity of coronal caries among older US adults and document their need for prevention interventions, we systematically reviewed studies on coronal caries incidence, increment, and attack rate. We abstracted six studies and calculated summary measures using a random-effects model (95% confidence interval [95%CI]). We tested for heterogeneity and identified associated factors by examining the correlation between outcome measures and baseline population risk and study characteristics. We re-calculated summary measures after adjusting outcomes that netted out examiner reversals. Incidence and increment varied significantly by study. Adjusting studies for netting out examiner reversals reduced heterogeneity significantly. Annual attack rate among adjusted North American studies was 1.4 surfaces per 100 surfaces (95%CI = 1.0 1.9), or approximately 1 new carious surface per person per year. These rates are equal to or higher than those in children and indicate a need for caries prevention services. PMID- 16040729 TI - Efficacy of silver diamine fluoride for caries reduction in primary teeth and first permanent molars of schoolchildren: 36-month clinical trial. AB - We hypothesized that the six-monthly application of silver diamine fluoride (SDF) can arrest the development of caries in the deciduous dentition of six-year-old schoolchildren and prevent caries in their first permanent molars. A prospective controlled clinical trial was conducted on the efficacy of a 38% SDF solution for caries reduction. Four hundred and twenty-five six-year-old children were divided into two groups: One group received SDF solution in primary canines and molars and first permanent molars every 6 mos for 36 mos. The second group served as controls. The 36-month follow-up was completed by 373 children. The mean number of new decayed surfaces appearing in primary teeth during the study was 0.29 in the SDF group vs. 1.43 in controls. The mean of new decayed surfaces in first permanent molars was 0.37 in the SDF group vs. 1.06 in controls. The SDF solution was found to be effective for caries reduction in primary teeth and first permanent molars in schoolchildren. PMID- 16040730 TI - Oxygen inhibition in dental resins. AB - Oxygen inhibits free radical polymerization and yields polymers with uncured surfaces. This is a concern when thin layers of resin are being polymerized, or in circumstances where conventional means of eliminating inhibition are inappropriate. In this study, we tested the hypothesis that viscosity, filler content, and polymerization temperature modify oxygen diffusion in the resin or the reactivity of radical species, and affect the degree of conversion near the surface. Confocal Raman micro-spectroscopy was used to measure monomer conversion from the surface to the bulk of cured resins. Increased viscosity was shown to limit oxygen diffusion and increase conversion near the surface, without necessarily modifying the depth of inhibition. The filler material was shown to increase, simultaneously, oxygen diffusivity and the viscosity of the resin, which have opposite effects on conversion. Polymerization at a temperature above approximately 110 degrees C was shown to eliminate oxygen inhibition. PMID- 16040731 TI - Incompatibility of oxalate desensitizers with acidic, fluoride-containing total etch adhesives. AB - The use of oxalate desensitizers on acid-etched dentin prior to adhesive application can result in subsurface tubular occlusion by calcium oxalate crystals. However, the solubility of calcium oxalate increases in acidic solution. We hypothesized that total-etch adhesives can, depending upon their pH, interact with oxalate-desensitizer-treated dentin in an adverse manner. Acid etched human dentin treated with 2 oxalate desensitizers (BisBlock and Super Seal) was bonded with 4 simplified total-etch adhesives: One-Step (OS), Single Bond (SB), OptiBond Solo Plus (OB), and Prime&Bond NT (PB). Composite-dentin beams were examined by SEM and TEM, both of which revealed numerous spherical globules on OB- and PB-bonded, desensitizer-treated dentin, but not in OS or SB samples. Bond strengths produced by OB and PB were significantly lower in oxalate treated specimens than those produced by OS or SB. These surface globules may have interfered with hybridization of demineralized dentin with OB and PB resins and caused compromised bond strengths. PMID- 16040732 TI - EDTA treatment improves resin-dentin bonds' resistance to degradation. AB - The existence of unprotected collagen fibrils within the hybrid layer compromises the longevity of restorations. This phenomenon may be avoided if solutions other than strong acids are used for dentin demineralization. The hypothesis to be tested is that bond durability may be improved by EDTA demineralization. Dentin surfaces (human and bovine) were bonded: (1) after phosphoric-acid-etching, and after EDTA demineralization with (2) a total-etch adhesive and (3) a self-etching adhesive. After the teeth were sectioned into beams, half of the specimens were immersed in NaOCl, while the other half was immersed in water. Beams were tested to failure in tension. ANOVA and multiple-comparisons tests were used (P < 0.05). No differences in bond strength were found among the 3 bonding procedures, although bonds made to human molars were 43-61% higher than those to bovine incisors. After NaOCl immersion, only specimens subjected to EDTA demineralization maintained the initial bond strength. We conclude that the collagen network is better-preserved after EDTA demineralization. PMID- 16040733 TI - Chlorhexidine arrests subclinical degradation of dentin hybrid layers in vivo. AB - The recent paradigm that endogenous collagenolytic and gelatinolytic activities derived from acid-etched dentin result in degradation of hybrid layers requires in vivo validation. This study tested the null hypothesis that there is no difference between the degradation of dentin bonded with an etch-and-rinse adhesive and that in conjunction with chlorhexidine, an MMP inhibitor, applied after phosphoric-acid-etching. Contralateral pairs of bonded Class I restorations in primary molars of clinical subjects were retrieved after a six-month period of intra-oral functioning and processed for transmission electron microscopy. Hybrid layers from the chlorhexidine-treated teeth exhibited normal structural integrity of the collagen network. Conversely, abnormal hybrid layers were seen in the control teeth, with progressive disintegration of the fibrillar network, to the extent that it was beyond detection by collagen staining. Self-destruction of collagen matrices occurs rapidly in resin-infiltrated dentin in vivo and may be arrested with the use of chlorhexidine as an MMP inhibitor. PMID- 16040734 TI - Cyclosporin A specifically affects nuclear PLCbeta1 in immunodepressed heart transplant patients with gingival overgrowth. AB - One of the most commonly observed adverse effects of cyclosporin A (CsA) is the development of gingival overgrowth (GO). Fibroblasts are involved in GO, but the question why only a percentage of patients undergoing CsA treatment shows this side-effect remains unanswered. In a previous study, CsA has been demonstrated to induce over-expression of phospholipase C (PLC) beta(1) in fibroblasts of patients with clinical GO, in cells from both enlarged and clinically healthy gingival sites. In this work, we assessed the expression of PLCbeta isoforms to investigate whether the exaggerated fibroblast response to CsA related to increased PLCbeta(1) expression could also be detected in CsA-treated patients without clinical signs of GO. Our results support the hypothesis of a multi factorial origin of gingival overgrowth, including specific changes within the gingival tissues orchestrating fibroblastic hyper-responsiveness as a consequence of a long-term in vivo exposure to cyclosporin A. PMID- 16040735 TI - Effects of a high-cholesterol diet on cell behavior in rat periodontitis. AB - Studies have shown an association between periodontitis and serum cholesterol levels. We hypothesized that high dietary cholesterol could influence periodontitis as a result of proliferation of the junctional epithelium. Rats were divided into 4 groups. Two groups were fed a regular diet, and 2 groups were fed a high-cholesterol diet. One of each dietary group was treated with periodontitis-inducing agents (lipopolysaccharide and proteases), while the other was treated with pyrogen-free water. Feeding rats with a high-cholesterol diet induced an increase in blood total cholesterol and a decrease in high-density lipoprotein cholesterol. Proliferation of the junctional epithelium with increasing bone resorption was promoted by the consumption of a high-cholesterol diet. High dietary cholesterol further increased the cell-proliferative activity of the junctional epithelium induced by lipopolysaccharide and proteases. These results suggest that high dietary cholesterol can initiate and augment periodontitis in the rat periodontitis model. PMID- 16040736 TI - Distraction osteogenesis may promote periodontal bone regeneration. AB - Distraction osteogenesis has numerous applications in the treatment of conditions affecting the dentofacial complex, from midface advancement to orthodontic tooth movement produced by transversal distraction of the periodontal ligament. The purpose of this study was to test the hypothesis that periodontium can be consistently distracted toward the tooth crown to promote periodontal bone regeneration. After the surgical production of periodontal defects in maxillary canines of 5 mongrel dogs, periodontal bone distraction was performed. Light microscopy was used for histopathological and morphometric analysis. Periodontal bone regeneration occurred in all animals. Periodontal bone regeneration in the distraction sites (Mean +/- SD: 5.45 +/- 2.01 mm) differed from that in control sites (0.008 +/- 0.67 mm; p < 0.0001). Periodontal bone distraction resulted in periodontal bone regeneration. This finding may establish periodontal bone distraction as a new treatment alternative for periodontal defects. PMID- 16040737 TI - Effect of NOS inhibitor on cytokine and COX2 expression in rat pulpitis. AB - Various kinds of chemical mediators are synthesized in the course of pulpitis; thus, control of their production would assist in inducing a reduction in pulpal inflammation. We hypothesized that nitric oxide (NO) would be an important mediator of pulpal inflammation. Pulpal inflammation was induced by the application of LPS in rat incisor pulp, and inducible nitric oxide synthase (iNOS) expression was evaluated by reverse-transcription/polymerase chain reaction and immunohistochemical staining. After LPS application, iNOS mRNA was first detected after 3 hrs, peaked at 6 hrs, and decreased thereafter. iNOS positive cells were macrophages and neutrophils. An NOS inhibitor caused drastic decreases in the expression of pro-inflammatory cytokines and COX2 mRNA, which was highly induced in the LPS-induced pulpitis. These results indicate that NO synthesis is related to the initiation of mediator production, and that its down regulation should contribute to the prevention of pro-inflammatory mediator synthesis. PMID- 16040738 TI - Characterization of nonsymbiotic tomato hemoglobin. AB - The nonsymbiotic tomato hemoglobin SOLly GLB1 (Solanum lycopersicon) is shown to form a homodimer of approximately 36 kDa with a high affinity for oxygen. Furthermore, our combined ultraviolet/visible, resonance Raman, and continuous wave electron paramagnetic resonance (EPR) measurements reveal that a mixture of penta- and hexacoordination of the heme iron is found in the deoxy ferrous form, whereas the ferric form shows predominantly a bis-histidine ligation (F8His Fe(2+/3+)-E7His). This differs from the known forms of vertebrate hemoglobins and myoglobins. We have successfully applied our recently designed pulsed-EPR strategy to study the low-spin ferric form of tomato hemoglobin. These experiments reveal that, in ferric SOLly GLB1, one of the histidine planes is rotated 20 degrees (+/-10 degrees ) away from a N(heme)-Fe-N(heme) axis. Additionally, the observed g-values indicate a quasicoplanarity of the histidine ligands. From the HYSCORE (hyperfine sublevel correlation) measurements, the hyperfine and nuclear quadrupole couplings of the heme and histidine nitrogens are identified and compared with known EPR/ENDOR data of vertebrate Hbs and cytochromes. Finally, the ligand binding kinetics, which also indicate that the ferrous tomato Hb is only partially hexacoordinated, will be discussed in relation with the heme-pocket structure. The similarities and differences with other known nonsymbiotic plant hemoglobins will be highlighted. PMID- 16040739 TI - Noise and functional protein dynamics. AB - The magnetic field dependence of the proton-spin-lattice relaxation rate in rotationally immobilized proteins shows that the one-dimensional character of the protein primary structure causes a dramatic increase in the population of low frequency motions from 10 kHz to 20 MHz. As a consequence, the probability and rate at which functionally critical conformational states are thermally sampled in a protein are dramatically increased as well, when compared with a three dimensional lattice structure. Studies of protein dynamics often focus on time periods far shorter than those associated with catalytic function, but we show here that the magnetic field dependence of the proton nuclear spin-lattice relaxation rate in rotationally immobilized proteins reports unambiguously the structural fluctuations in the frequency range from 10 kHz to 20 MHz. This relaxation rate decreases with increasing Larmor frequency according to a power law that derives from the distribution of dynamical states, the localization of the structural disturbances, and the spatial distribution of hydrogen atoms in the structure. The robust theoretical foundation for the spin-relaxation process, loosely characterized as a direct spin-phonon coupling, shows that the disturbances propagate in a space of reduced dimensionality, essentially along the stiff connections of the polypeptide chain. The reduced dimensionality traps the disturbance and changes the efficiency for energy redistribution in the protein and the processes that drive nuclear spin relaxation. We also show that the Larmor frequency dependence of the protein-proton-spin-lattice relaxation rate constant is related to the frequency dependence of force constants and mean square displacement commonly observed or calculated for proteins. We believe that these approaches give additional physical insight into the character of the extremely low-frequency protein dynamics. PMID- 16040740 TI - Nuclear localization signal peptides induce molecular delivery along microtubules. AB - Many essential processes in eukaryotic cells depend on regulated molecular exchange between its two major compartments, the cytoplasm and the nucleus. In general, nuclear import of macromolecular complexes is dependent on specific peptide signals and their recognition by receptors that mediate translocation through the nuclear pores. Here we address the question of how protein products bearing such nuclear localization signals arrive at the nuclear membrane before import, i.e., by simple diffusion or perhaps with assistance of cytoskeletal elements or cytoskeleton-associated motor proteins. Using direct single-particle tracking and detailed statistical analysis, we show that the presence of nuclear localization signals invokes active transport along microtubules in a cell-free Xenopus egg extract. Chemical and antibody inhibition of minus-end directed cytoplasmic dynein blocks this active movement. In the intact cell, where microtubules project radially from the centrosome, such an interaction would effectively deliver nuclear-targeted cargo to the nuclear envelope in preparation for import. PMID- 16040741 TI - Role of voltage-dependent modulation of store Ca2+ release in synchronization of Ca2+ oscillations. AB - Slow waves are rhythmic depolarizations that underlie mechanical activity of many smooth muscles. Slow waves result through rhythmic Ca(2+) release from intracellular Ca(2+) stores through inositol 1,4,5-trisphosphate (IP(3)) sensitive receptors and Ca(2+)-induced Ca(2+) release. Ca(2+) oscillations are transformed into membrane depolarizations by generation of a Ca(2+)-activated inward current. Importantly, the store Ca(2+) oscillations that underlie slow waves are entrained across many cells over large distances. It has been shown that IP(3) receptor-mediated Ca(2+) release is enhanced by membrane depolarization. Previous studies have implicated diffusion of Ca(2+) or the second messenger IP(3) across gap junctions in synchronization of Ca(2+) oscillations. In this study, a novel mechanism of Ca(2+) store entrainment through depolarization-induced IP(3) receptor-mediated Ca(2+) release is investigated. This mechanism is significantly different from chemical coupling based mechanisms, as membrane potential has a coupling effect over distances several orders of magnitude greater than either diffusion of Ca(2+) or IP(3) through gap junctions. It is shown that electrical coupling acting through voltage-dependent modulation of store Ca(2+) release is able to synchronize oscillations of cells even when cells are widely separated and have different intrinsic frequencies of oscillation. PMID- 16040742 TI - Molecular dynamics simulation studies of the effect of phosphocitrate on crystal induced membranolysis. AB - In this study, following our earlier work on calcium pyrophosphate dihydrate (CPPD) crystal-induced membranolysis, we demonstrate, using the CHARMM method of molecular dynamics simulation, the protective role of phosphocitrate (PC) against solvated dimyristoyl phosphatidylcholine phospholipid bilayer disintegration on contact with the CPPD crystal. Our molecular dynamics simulations studies show that coverage of the CPPD crystal with a layer of phosphocitrate molecules results in the conservation of phospholipid bilayer integrity. We show that the rupture of the lipid bilayer in presence of CPPD and the protective effect of PC are primarily due to electrostatic interactions. The protective role of PC, which may also play an important and potentially therapeutic function against crystal induced membranolysis is also discussed. PMID- 16040743 TI - The amino-terminal PrP domain is crucial to modulate prion misfolding and aggregation. AB - The main hypothesis for prion diseases is that the cellular protein (PrP(C)) can be altered into a misfolded, beta-sheet-rich isoform (PrP(Sc)), which undergoes aggregation and triggers the onset of transmissible spongiform encephalopathies. Here, we investigate the effects of amino-terminal deletion mutations, rPrP(Delta51-90) and rPrP(Delta32-121), on the stability and the packing properties of recombinant murine PrP. The region lacking in rPrP(Delta51-90) is involved physiologically in copper binding and the other construct lacks more amino-terminal residues (from 32 to 121). The pressure stability is dramatically reduced with decreasing N-domain length and the process is not reversible for rPrP(Delta51-90) and rPrP(Delta32-121), whereas it is completely reversible for the wild-type form. Decompression to atmospheric pressure triggers immediate aggregation for the mutants in contrast to a slow aggregation process for the wild-type, as observed by Fourier-transform infrared spectroscopy. The temperature-induced transition leads to aggregation of all rPrPs, but the unfolding temperature is lower for the rPrP amino-terminal deletion mutants. The higher susceptibility to pressure of the amino-terminal deletion mutants can be explained by a change in hydration and cavity distribution. Taken together, our results show that the amino-terminal region has a pivotal role on the development of prion misfolding and aggregation. PMID- 16040744 TI - Quantitative multiphoton spectral imaging and its use for measuring resonance energy transfer. AB - Protein labeling with green fluorescent protein derivatives has become an invaluable tool in cell biology. Protein quantification, however, is difficult when cells express constructs with overlapping fluorescent emissions. Under these conditions, signal separation using emission filters is inherently inefficient. Spectral imaging solves this problem by recording emission spectra directly. Unfortunately, linear unmixing, the algorithm used for quantifying individual fluorophores from emission spectra, fails when resonance energy transfer (RET) is present. We therefore sought to develop an unmixing algorithm that incorporates RET. An equation for spectral emission incorporating RET was derived and an assay based on this formalism, spectral RET (sRET), was developed. Standards with defined RET efficiencies and with known Cerulean/Venus ratios were constructed and used to test sRET. We demonstrate that sRET analysis is a comprehensive, photon-efficient method for imaging RET efficiencies and accurately determines donor and acceptor concentrations in living cells. PMID- 16040745 TI - Absorption spectra of photoactive yellow protein chromophores in vacuum. AB - The absorption spectra of two photoactive yellow protein model chromophores have been measured in vacuum using an electrostatic ion storage ring. The absorption spectrum of the isolated chromophore is an important reference for deducing the influence of the protein environment on the electronic energy levels of the chromophore and separating the intrinsic properties of the chromophore from properties induced by the protein environment. In vacuum the deprotonated trans thiophenyl-p-coumarate model chromophore has an absorption maximum at 460 nm, whereas the photoactive yellow protein absorbs maximally at 446 nm. The protein environment thus only slightly blue-shifts the absorption. In contrast, the absorption of the model chromophore in aqueous solution is significantly blue shifted (lambda(max) = 395 nm). A deprotonated trans-p-coumaric acid has also been studied to elucidate the effect of thioester formation and phenol deprotonation. The sum of these two changes on the chromophore induces a red shift both in vacuum and in aqueous solution. PMID- 16040746 TI - Dynamical mechanisms of pacemaker generation in IK1-downregulated human ventricular myocytes: insights from bifurcation analyses of a mathematical model. AB - Dynamical mechanisms of the biological pacemaker (BP) generation in human ventricular myocytes were investigated by bifurcation analyses of a mathematical model. Equilibrium points (EPs), periodic orbits, stability of EPs, and bifurcation points were determined as functions of bifurcation parameters, such as the maximum conductance of inward-rectifier K+ current (I(K1)), for constructing bifurcation diagrams. Stable limit cycles (BP activity) abruptly appeared around an unstable EP via a saddle-node bifurcation when I(K1) was suppressed by 84.6%. After the bifurcation at which a stable EP disappears, the I(K1)-reduced system has an unstable EP only, which is essentially important for stable pacemaking. To elucidate how individual sarcolemmal currents contribute to EP instability and BP generation, we further explored the bifurcation structures of the system during changes in L-type Ca2+ channel current (I(Ca,L)), delayed rectifier K+ currents (I(K)), or Na(+)/Ca2+ exchanger current (I(NaCa)). Our results suggest that 1), I(Ca,L) is, but I(K) or I(NaCa) is not, responsible for EP instability as a requisite to stable BP generation; 2), I(K) is indispensable for robust pacemaking with large amplitude, high upstroke velocity, and stable frequency; and 3), I(NaCa) is the dominant pacemaker current but is not necessarily required for the generation of spontaneous oscillations. PMID- 16040747 TI - Anesthetic interaction with ketosteroid isomerase: insights from molecular dynamics simulations. AB - The nature and the sites of interactions between anesthetic halothane and homodimeric Delta5-3-ketosteroid isomerase (KSI) are characterized by flexible ligand docking and confirmed by 1H-15N NMR. The dynamics consequence of halothane interaction and the implication of the dynamic changes to KSI function are studied by multiple 5-ns molecular dynamics simulations in the presence and absence of halothane. Both docking and MD simulations show that halothane prefer the amphiphilic dimeric interface to the hydrophobic active site of KSI. Halothane occupancy at the dimer interface disrupted the intersubunit hydrogen bonding formed either directly through side chains of polar residues or indirectly through the mediation of the interfacial water molecules. Moreover, in the presence of halothane, the exchange rate of the bound waters with bulk water was increased. Halothane perturbation to the dimer interface affected the overall flexibility of the active site. This action is likely to contribute to the halothane-induced reduction of the KSI activity. The allosteric halothane modulation of the dynamics-function relationship of KSI without direct competition at the enzymatic active sites may be generalized to offer a unifying explanation of anesthetic action on a diverse range of multidomain neuronal proteins that are potentially relevant to clinical general anesthesia. PMID- 16040748 TI - Interaction of the antimicrobial peptide cyclo(RRWWRF) with membranes by molecular dynamics simulations. AB - Antimicrobial peptides have gained a lot of interest in recent years due to their potential use as a new generation of antibiotics. It is believed that this type of relatively short, amphipathic, cationic peptide targets the bacterial membrane, and destroys the chemical gradients over the membrane via formation of stable or transient pores. Here we use the NMR structure of cyclo(RRWWRF) in a series of molecular dynamics simulations in membranes at various peptide/lipid ratios. We observe that the NMR structure of the peptide is still stable after 100 ns simulation. At a peptide/lipid ratio of 2:128, the membrane is only a little affected compared to a pure dipalmitoylphosphatidylcholine lipid membrane, but at a ratio of 12:128, the water-lipid interface becomes more fuzzy, the water molecules can reach deeper into the hydrophobic core, and the water penetration free-energy barrier changes. Moreover, we observe that the area per lipid decreases and the deuterium order parameters increase in the presence of the peptide. We suggest that the changes in the hydrophobic core, together with the changes in the headgroups, result in an imbalance of the membrane and that it is thus not an efficient hydrophobic barrier in the presence of the peptides, independent of pore formation. PMID- 16040749 TI - Coarse-grained strategy for modeling protein stability in concentrated solutions. AB - We present a coarse-grained approach for modeling the thermodynamic stability of single-domain globular proteins in concentrated aqueous solutions. Our treatment derives effective protein-protein interactions from basic structural and energetic characteristics of the native and denatured states. These characteristics, along with the intrinsic (i.e., infinite dilution) thermodynamics of folding, are calculated from elementary sequence information using a heteropolymer collapse theory. We integrate this information into Reactive Canonical Monte Carlo simulations to investigate the connections between protein sequence hydrophobicity, protein-protein interactions, protein concentration, and the thermodynamic stability of the native state. The model predicts that sequence hydrophobicity can affect how protein concentration impacts native-state stability in solution. In particular, low hydrophobicity proteins are primarily stabilized by increases in protein concentration, whereas high hydrophobicity proteins exhibit richer nonmonotonic behavior. These trends appear qualitatively consistent with the available experimental data. Although factors such as pH, salt concentration, and protein charge are also important for protein stability, our analysis suggests that some of the nontrivial experimental trends may be driven by a competition between destabilizing hydrophobic protein protein attractions and entropic crowding effects. PMID- 16040750 TI - Multi-step fibrinogen binding to the integrin (alpha)IIb(beta)3 detected using force spectroscopy. AB - The regulated ability of integrin alphaIIbbeta3 to bind fibrinogen plays a crucial role in platelet aggregation and hemostasis. We have developed a model system based on laser tweezers, enabling us to measure specific rupture forces needed to separate single receptor-ligand complexes. First of all, we performed a thorough and statistically representative analysis of nonspecific protein-protein binding versus specific alphaIIbbeta3-fibrinogen interactions in combination with experimental evidence for single-molecule measurements. The rupture force distribution of purified alphaIIbbeta3 and fibrinogen, covalently attached to underlying surfaces, ranged from approximately 20 to 150 pN. This distribution could be fit with a sum of an exponential curve for weak to moderate (20-60 pN) forces, and a Gaussian curve for strong (>60 pN) rupture forces that peaked at 80 90 pN. The interactions corresponding to these rupture force regimes differed in their susceptibility to alphaIIbbeta3 antagonists or Mn2+, an alphaIIbbeta3 activator. Varying the surface density of fibrinogen changed the total binding probability linearly >3.5-fold but did not affect the shape of the rupture force distribution, indicating that the measurements represent single-molecule binding. The yield strength of alphaIIbbeta3-fibrinogen interactions was independent of the loading rate (160-16,000 pN/s), whereas their binding probability markedly correlated with the duration of contact. The aggregate of data provides evidence for complex multi-step binding/unbinding pathways of alphaIIbbeta3 and fibrinogen revealed at the single-molecule level. PMID- 16040751 TI - DNA cholesteric pitch as a function of density and ionic strength. AB - The nature of chiral interactions among chiral biopolymers, such as DNA, protein alpha-helices, and rodlike virus particles, remains elusive. In particular, a satisfactory model connecting molecular chiral interactions and the pitch of the resulting chiral mesophases is lacking. We report the measurement of short fragment (146-bp) DNA cholesteric spherulite pitch as a function of osmotic pressure, average DNA interaxial spacing, and salt concentration. We determined cholesteric pitch and interaxial spacing by polarizing optical microscopy and x ray scattering, respectively, from which the twist-angle between DNA molecules can be calculated. Surprisingly, we found that decreasing ionic strength resulted in weaker chiral interactions between DNA chains, as evidenced by the decrease in the twist-angle, and consequent increase in the cholesteric pitch, for a fixed interaxial spacing. We propose that this behavior can be explained by increased smearing-out of the helical charge pattern along DNA as the Debye screening length is increased. PMID- 16040752 TI - Statistical kinetics of macromolecular dynamics. AB - Fluctuations in biochemical processes can provide insights into the underlying kinetics beyond what can be gleaned from studies of average rates alone. Historically, analysis of fluctuating transmembrane currents supplied information about ion channel conductance states and lifetimes before single-channel recording techniques emerged. More recently, fluctuation analysis has helped to define mechanochemical pathways and coupling ratios for the motor protein kinesin as well as to probe the contributions of static and dynamic disorder to the kinetics of single enzymes. As growing numbers of assays are developed for enzymatic or folding behaviors of single macromolecules, the range of applications for fluctuation analysis increases. To evaluate specific biochemical models against experimental data, one needs to predict analytically the distribution of times required for completion of each reaction pathway. Unfortunately, using traditional methods, such calculations can be challenging for pathways of even modest complexity. Here, we derive an exact expression for the distribution of completion times for an arbitrary pathway with a finite number of states, using a recursive method to solve algebraically for the appropriate moment-generating function. To facilitate comparisons with experiments on processive motor proteins, we develop a theoretical formalism for the randomness parameter, a dimensionless measure of the variance in motor output. We derive the randomness for motors that take steps of variable sizes or that move on heterogeneous substrates, and then discuss possible applications to enzymes such as RNA polymerase, which transcribes varying DNA sequences, and to myosin V and cytoplasmic dynein, which may advance by variable increments. PMID- 16040753 TI - Electronic transport in DNA. AB - We study the electronic properties of DNA by way of a tight-binding model applied to four particular DNA sequences. The charge transfer properties are presented in terms of localization lengths (crudely speaking, the length over which electrons travel). Various types of disorder, including random potentials, are employed to account for different real environments. We have performed calculations on poly(dG)-poly(dC), telomeric-DNA, random-ATGC DNA, and lambda-DNA. We find that random and lambda-DNA have localization lengths allowing for electron motion among a few dozen basepairs only. A novel enhancement of localization lengths is observed at particular energies for an increasing binary backbone disorder. We comment on the possible biological relevance of sequence-dependent charge transfer in DNA. PMID- 16040754 TI - Nodal cilia dynamics and the specification of the left/right axis in early vertebrate embryo development. AB - Nodal cilia dynamics is a key factor for left/right axis determination in mouse embryos through the induction of a leftward fluid flow. So far it has not been clearly established how such dynamics is able to induce the asymmetric leftward flow within the node. Herein we propose that an asymmetric two-phase nonplanar beating cilia dynamics that involves the bending of the ciliar axoneme is responsible for the leftward fluid flow. We support our proposal with a host of hydrodynamic arguments, in silico experiments and in vivo video microscopy data in wild-type embryos and inv mutants. Our phenomenological modeling approach underscores how the asymmetry and speed of the flow depends on different relevant parameters. In addition, we discuss how the combination of internal and external mechanisms might cause the two-phase beating cilia dynamics. PMID- 16040755 TI - The conserved N-terminal region of the mitotic checkpoint protein BUBR1: a putative TPR motif of high surface activity. AB - BUBR1, a key component of the mitotic spindle checkpoint, is a multidomain protein kinase that is activated in response to kinetochore tension. Although BUB1 and BUBR1 play an important role in cell division, very little is known about their structural characteristics. We show that the conserved N-terminal region of BUBR1, comprising residues 1-204, is a globular domain of high alpha helical content ( approximately 60%), stable in the pH range 4-9 and probably organized as a tetratricopeptide motif repeat (TPR), most closely resembling residues 16-181 of protein phosphatase 5. Because the latter presents a continuous amphipathic groove and is regulated by binding certain fatty acids, we compared the properties of BUBR1(1-204) and TPR-PP5(16-181) at air/water interfaces and found that both proteins exhibited a similar surface activity and formed stable, rigid monolayers. The deletion of a region that probably comprises several alpha-helices of BUBR1 indicates that long-range interactions are essential for the stability of the N-terminal domain. The presence of the putative TPR motif strongly suggests that the N-terminal domain of BUBR1 is involved in direct protein-protein interactions and/or protein-lipid interactions. PMID- 16040756 TI - Quantitative modeling of chloride conductance in yeast TRK potassium transporters. AB - So-called TRK proteins are responsible for active accumulation of potassium in plants, fungi, and bacteria. A pair of these proteins in the plasma membrane of Saccharomyces cerevisiae, ScTrk1p and ScTrk2p, also admit large, adventitious, chloride currents during patch-recording (Cl- efflux). Resulting steady-state current-voltage curves can be described by two simple kinetic models, most interestingly, voltage-driven channeling of ions through a pair of activation energy barriers that lie within the membrane dielectric, near the inner (alpha) and outer (beta) surfaces. Two barrier heights (E(alpha) and E(beta)) and two relative distances (a1 and b2) from the surfaces specify the model. Measured current amplitude parallels intracellular chloride concentration and is strongly enhanced by acidic extracellular pH. The former implies an exponential variation of a1, between approximately 0.2 and approximately 0.4 of the membrane thickness, whereas the latter implies a linear variation of E(beta), by 0.69 Kcal mol( 1)/pH. The model requires membrane slope conductance to rise exponentially with increasingly large negative membrane voltage, as verified by data from a few yeast spheroplasts that tolerated voltage clamping at -200 to -300 mV. The behaviors of E(beta) and a1 accord qualitatively with a hypothetical structural model for fungal TRK proteins, suggesting that chloride ions flow through a central pore formed by symmetric aggregation of four TRK monomers. PMID- 16040757 TI - A computational study of the closed and open states of the influenza a M2 proton channel. AB - In this study, four possible conformations of the His-37 and Trp-41 residues for the closed state of the influenza M2 ion channel were identified by a conformation scan based on a solid-state NMR restraint. In the four conformations, the His-37 residue can be of either the t-160 or t60 rotamer, whereas Trp-41 can be of either the t-105 or t90 rotamer. These conformations were further analyzed by density functional theory calculations and molecular dynamics simulations, and the data indicate that the His-37 residue most likely adopts the t60 rotamer and should be monoprotonated at the delta-nitrogen site, whereas Trp-41 adopts the t90 rotamer. This result is consistent with published experimental data and points to a simple gating mechanism: in the closed state, the His-37 and Trp-41 residues adopt the (t60, t90) conformation, which nearly occludes the pore, preventing nonproton ions from passing through due to the steric and desolvation effects. Moreover, the His-37 tetrad interrupts the otherwise continuous hydrogen-bonding network of the pore water by forcing the water molecules above and below it to adopt opposite orientations, thus adding to the blockage of proton shuttling. The channel can be easily opened by rotating the His-37 chi2 angle from 60 to 0 degrees . This open structure allows pore water to penetrate the constrictive region and to form a continuous water wire for protons to shuttle through, while being still narrow enough to exclude other ions. PMID- 16040758 TI - Caveolin-1 expression and membrane cholesterol content modulate N-type calcium channel activity in NG108-15 cells. AB - Caveolins are the main structural proteins of glycolipid/cholesterol-rich plasmalemmal invaginations, termed caveolae. In addition, caveolin-1 isoform takes part in membrane remodelling as it binds and transports newly synthesized cholesterol from endoplasmic reticulum to the plasma membrane. Caveolin-1 is expressed in many cell types, including hippocampal neurons, where an abundant SNAP25-caveolin-1 complex is detected after induction of persistent synaptic potentiation. To ascertain whether caveolin-1 influences neuronal voltage-gated Ca2+ channel basal activity, we stably expressed caveolin-1 into transfected neuroblastoma x glioma NG108-15 hybrid cells [cav1(+) clone] that lack endogenous caveolins but express N-type Ca2+ channels upon cAMP-induced neuronal differentiation. Whole-cell patch-clamp recordings of cav1(+) cells demonstrated that N-type current density was reduced in size by approximately 70% without any significant change in the time course of activation and inactivation and voltage dependence. Moreover, the cav1(+) clone exhibited a significantly increased proportion of membrane cholesterol compared to wild-type NG108-15 cells. To gain insight into the mechanism underlying caveolin-1 lowering of N-current density, and more precisely to test whether this was indirectly caused by caveolin-1 induced enhancement of membrane cholesterol, we compared single N-type channel activities in cav1(+) clone and wild-type NG108-15 cells enriched with cholesterol after exposure to a methyl-beta-cyclodextrin-cholesterol complex. A lower Ca2+ channel activity was recorded from cell-attached patches of both cell types, thus supporting the view that the increased proportion of membrane cholesterol is ultimately responsible for the effect. This is due to a reduction in the probability of channel opening caused by a significant decrease of channel mean open time and by an increase of the frequency of null sweeps. PMID- 16040759 TI - Substrate-supported phospholipid membranes studied by surface plasmon resonance and surface plasmon fluorescence spectroscopy. AB - Substrate-supported planar lipid bilayer membranes are attractive model cellular membranes for biotechnological applications such as biochips and sensors. However, reliable fabrication of the lipid membranes on solid surfaces still poses significant technological challenges. In this study, simultaneous surface plasmon resonance (SPR) and surface plasmon fluorescence spectroscopy (SPFS) measurements were applied to the monitoring of adsorption and subsequent reorganization of phospholipid vesicles on solid substrates. The fluorescence intensity of SPFS depends very sensitively on the distance between the gold substrate and the fluorophore because of the excitation energy transfer to gold. By utilizing this distance dependency, we could obtain information about the topography of the adsorbed membranes: Adsorbed vesicles could be clearly distinguished from planar bilayers due to the high fluorescence intensity. SPSF can also incorporate various analytical techniques to evaluate the physicochemical properties of the adsorbed membranes. As an example, we demonstrated that the lateral mobility of lipid molecules could be estimated by observing the recovery of fluorescence after photobleaching. Combined with the film thickness information obtained by SPR, SPR-SPFS proved to be a highly informative technique to monitor the lipid membrane assembly processes on solid substrates. PMID- 16040760 TI - Positively charged surfaces increase the flexibility of DNA. AB - Many proteins "bind" DNA through positively charged amino acids on their surfaces. However, to overcome significant energetic and topological obstacles, proteins that bend or package DNA might also modulate the stiffness that is generated by repulsions between phosphates within DNA. Much previous work describes how ions change the flexibility of DNA in solution, but when considering macromolecules such as chromatin in which the DNA contacts the nucleosome core each turn of the double helix, it may be more appropriate to assess the flexibility of DNA on charged surfaces. Mica coated with positively charged molecules is a convenient substrate upon which the flexibility of DNA may be directly measured with a scanning force microscope. In the experiments described below, the flexibility of DNA increased as much as fivefold depending on the concentration and type of polyamine used to coat mica. Using theory that relates charge neutralization to flexibility, we predict that phosphate repulsions were attenuated by approximately 50% in the most flexible DNA observed. This simple method is an important tool for investigating the physiochemical causes and molecular biological effects of DNA flexibility, which affects DNA biochemistry ranging from chromatin stability to viral encapsulation. PMID- 16040761 TI - Kinetics of Ca2+ binding to the SR Ca-ATPase in the E1 state. AB - The time-resolved kinetics of Ca2+ binding to the SR Ca-ATPase in the E1 state was investigated by Ca(2+)-concentration jump experiments. Ca2+ was released by an ultraviolet-light flash from caged calcium, and charge movements in the membrane domain of the ion pumps were detected by the fluorescent styryl dye 2BITC. The partial reaction (H3E1 <-->) E1 <--> CaE1 <--> Ca2E1 can be characterized by two time constants, tau1 and tau2, both of which are not significantly Ca(2+)-concentration-dependent and only weakly pH-dependent at pH < 7.5. Both time constants differ by a factor of approximately 50 (4.7 vs. 200 ms). The weak substrate-dependence indicates that the rate-limiting process is not related to Ca2+ migration through the access channel and ion binding to the binding sites but to conformational rearrangements preceding the ion movements. The high activation energy obtained for both processes, 42.3 kJ mol(-1) and 60.3 kJ mol(-1) at pH 7.2, support this concept. Transient binding of Ca ions to the loop L67 and a movement of the Ca-loaded loop are discussed as a mechanism that facilitates the entrance of both Ca ions into the access channel to the ion binding sites. PMID- 16040762 TI - A critical reassessment of penetratin translocation across lipid membranes. AB - Penetratin is a short, basic cell-penetrating peptide able to induce cellular uptake of a vast variety of large, hydrophilic cargos. We have reassessed the highly controversial issue of direct permeation of the strongly cationic peptide across negatively charged lipid membranes. Confocal laser scanning microscopy on rhodamine-labeled giant vesicles incubated with carboxyfluorescein-labeled penetratin yielded no evidence of transbilayer movement, in contradiction to previously reported results. Confocal fluorescence spectroscopy on black lipid membranes confirmed this finding, which was also not affected by application of a transmembrane electric potential difference. A novel dialysis assay based on tryptophan absorbance and fluorescence spectroscopy demonstrated that the permeability of small and large unilamellar vesicles to penetratin is <10(-13) m/s. Taken together, the results show that penetratin is not capable of overcoming model membrane systems irrespective of the bilayer curvature or the presence of a transmembrane voltage. Thus, direct translocation across the hydrophobic core of the plasma membrane cannot account for the efficient uptake of penetratin into live cells, which is in accord with recent in vitro studies underlining the importance of endocytosis in the internalization process of cationic cell-penetrating peptides. PMID- 16040763 TI - Terahertz time-domain spectroscopy of amino acids and polypeptides. AB - Frequency-dependent absorption coefficients and refractive indices of amino acids (glycine and l-alanine) and polypeptides (polyglycine and poly-l-alanine) in the wavenumber region from 7 to 55 cm(-1) were measured by terahertz time-domain spectroscopy. A vibrational band was observed at 45.5 cm(-1) for polyglycine, which was assigned as an interchain mode. The reduced absorption cross sections of the amino acids and polypeptides show power-law behavior. The exponents are different between the monomers and polymers, and those of the two polypeptides suggest that the time dependences of the total dipole moments are similar in the timescale of subpico- to picoseconds. PMID- 16040765 TI - Ill-health retirement: time for regulations? PMID- 16040764 TI - Measuring single-bond rupture forces using high electric fields in microfluidic channels and DNA oligomers as force tags. AB - The disruption force of specific biotin-streptavidin bonds was determined using DNA oligomers as force tags. Forces were generated by an electric field acting on a biotinylated fluorescently labeled DNA oligomer. DNA oligomers were immobilized via biotin-streptavidin bonds on the walls of microfluidic channels. Channel layout and fluid-based deposition process were designed to enable well-defined localized deposition of the oligomers in a narrow gap of the microchannel. Electric fields of up to 400 V/cm were applied and electric field induced desorption of DNA oligomers was observed. At T approximately 30 degrees C, field induced desorption of both a 12 mer as well as a 48 mer yielded a streptavidin biotin disruption force of 75 fN. Streptavidin-functionalized surfaces remained intact and could be reloaded with biotinylated oligomers. At approximately 20 degrees C, however, no field-induced unbinding of the oligomers was observed at electric field strength of up to 400 V/cm, indicating a significant temperature dependence of the bond strength. PMID- 16040767 TI - In the current era of evidence-based guidelines, do consensus-based guidelines still have a place? PMID- 16040768 TI - The involvement of occupational health services prior to ill-health retirement in NHS staff in Scotland and predictors of re-employment. AB - OBJECTIVE: To assess the process, causes and outcomes of retirement because of ill-health in NHS staff in Scotland. Particular areas to be investigated include the involvement of occupational health services, access to rehabilitation and redeployment, current health, whether working again and to identify predictors of re-employment. METHOD: An ill-health retirement (IHR) questionnaire was mailed to 863 NHS staff awarded IHR benefits by the Scottish Public Pensions Agency between April 1998 and March 2000. RESULTS: In all, 49% of the 863 postal questionnaires were returned. The most common reasons for retiring were diseases of the musculoskeletal system (38%) and mental disorders (21%). Seventy-one percent of the participants reported their ill-health was partly or completely work related and 29% not work related. Ninety-two percent of NHS staff had attended an occupational health department prior to IHR. Twenty-three percent of participants had no contact with their line manager during their illness prior to retiral. Eighteen percent of individuals were offered the opportunity of working part-time and 15% offered alternative work. Seventeen percent of participants have obtained other work. Predictors of re-employment after IHR were: medical condition, managerial responsibility, improvement of health, wanting to work again, occupation and age at retirement. CONCLUSION: This is the first comprehensive study investigating NHS staff experiences of IHR in Scotland. This study illustrates the need for improved support and rehabilitation for ill-health care workers and that there is the potential to reduce levels of ill-health retirement. PMID- 16040769 TI - Ill-health retirement among healthcare workers in the Southern Health Board of the Republic of Ireland. AB - AIM: To determine the incidence rates, trends and medical causes of ill-health retirement (IHR) among different occupational classes in the Southern Health Board (SHB). METHODS: The 14 702 permanent employees of the SHB were divided into six occupational classes based on socio-economic status and occupational demands. The occupational classes were compared for incidence rates of IHR, age at IHR, years of service and medical causes of IHR. The total group of employees was used as the standard for statistical comparison. Incidence rates were compared using standardized IHR ratios (SIHRRs). Medical causes were compared using proportional ill-health retirement ratios (PIHRRs). RESULTS: Three hundred and three employees were granted IHR from 1994 to 2000. The overall incidence rate of IHR was 2.9 per 1000 employees per annum. The highest SIHRRs occurred in male maintenance staff at 345 (CI: 221-513) and female support staff at 158 (CI: 123-201). With regard to age and years of service, IHR peaked at a time that coincided with enhancement to pension entitlements. The common causes of IHR were musculoskeletal disorder (38%), mental illness(17%), circulatory disorder (12%) and neoplasia (8%). PIHRRs did not vary significantly between the classes. CONCLUSION: IHR was more common among manual healthcare workers. The structure of the pension scheme appeared to influence the timing of IHR. Occupational class did not appear to influence the medical causes of IHR. PMID- 16040770 TI - Phytophotodermatitis in grounds operatives. AB - Occupational dermatoses account for a significant proportion of work-related morbidity. Phytophotodermatitis is an unusual work-related dermatosis, but should be considered in the differential diagnosis of such conditions. An outbreak of 'strimmer rash' is described in three grounds operatives. The identification, aetiology and management of the condition are discussed. PMID- 16040771 TI - Occupational eosinophilic bronchitis without asthma due to chloramine exposure. AB - A case is discussed of eosinophilic bronchial inflammation without asthma due to chloramine T (CLT) exposure in a nurse. She reported a non-productive chronic cough on contact with CLT during workshifts. She had negative results of skin prick testing to CLT. However, sensitisation to CLT was confirmed by the presence of specific anti-chloramine IgE. Airway responsiveness to histamine was normal before and after CLT challenge. Eosinophil proportion in sputum was increased at 6 and 24 h after CLT challenge. PMID- 16040772 TI - Classification of Work Ability Index among young employees. AB - BACKGROUND: The reference values of the Work Ability Index (WAI) are best known for people aged over 45 years. However, the WAI score is highly dependent on age and population-based reference values for young employees are needed. AIM: To present WAI scores calculated for a population-based sample of employees in their early 30s. METHODS: A self-administered questionnaire including the items of the WAI was used. The study included 3725 employees (2021 men and 1704 women) aged 31 from part of the Northern Finland Birth Cohort 1966 Study (NFBC-66) and who were working in a wide range of occupations in northern Finland or in the Helsinki region during 1997-1998. RESULTS: The WAI scores among young employees varied from 15 to 49 with a mean of 40.7 points (SD 4.2). The 15th percentile, median and 85th percentile of the WAI scores were 37, 41 and 45 points, respectively. The WAI scores were relatively high for both men and women. CONCLUSIONS: When classification of WAI scores into four work ability categories is used, the scores of employees in their early 30s should be evaluated separately from those of older age groups. For young employees work ability categories poor (7-36 points), moderate (37-40 points), good (41-44 points) and excellent (45-49 points) are suggested. PMID- 16040773 TI - The NRL precedent? PMID- 16040775 TI - Pabulum. PMID- 16040774 TI - Re: chemical, biological, radiological and nuclear terrorism: an introduction for occupational physicians. PMID- 16040776 TI - Back pain. PMID- 16040777 TI - Safer devices preventing occupational blood and body fluid exposures. PMID- 16040778 TI - Re: the interpretation of zinc protoporphyrin changes in lead intoxication. PMID- 16040779 TI - Safe inoculation of blood for liquid culture. PMID- 16040783 TI - Rehabilitation decreases exercise-induced oxidative stress in chronic obstructive pulmonary disease. AB - The effect of exercise at different intensities as well as the effect of intensive supervised pulmonary rehabilitation on oxidative stress were studied for chronic obstructive pulmonary disease (COPD). Eleven patients with COPD and 11 healthy age-matched control subjects performed a maximal and submaximal exercise cycle ergometry test at 60% of peak workload. Patients with COPD performed these tests before and after 8 wk of pulmonary rehabilitation. Measurements were done before, immediately after, and 4 h after both exercise tests. At rest, increased oxidative stress was observed in patients compared with control subjects, as measured by urinary malondialdehyde (MDA; p < 0.05) and hydrogen peroxide (H2O2) in breath condensate (p < 0.05). In healthy control subjects, a significant increase in urinary MDA was observed 4 h after both exercise tests (p = 0.05), whereas H2O2 significantly increased immediately after maximal exercise (p < 0.05). In patients with COPD, before rehabilitation, reactive oxygen species-induced DNA damage in peripheral blood mononuclear cells, urinary MDA, and plasma uric acid were significantly increased after both exercise tests (p < 0.05), whereas no significant increase was observed in plasma MDA. In contrast, exhaled H2O2 was only significantly increased after maximal exercise (p < 0.02). Although after rehabilitation peak workload was increased by 24%, a similar oxidative stress response was found. Remarkably, a decrease in reactive oxygen species-induced DNA damage was detected after exercise at submaximal intensity despite increased exercise duration of 73%. In summary, patients with COPD had increased pulmonary and systemic oxidative stress both at rest and induced by exercise. In addition, pulmonary rehabilitation increased exercise capacity and was associated with reduced exercise-induced oxidative stress. PMID- 16040784 TI - Lung function decline, chronic bronchitis, and occupational exposures in young adults. AB - RATIONALE: Occupational exposures to vapors, gas, dust, or fumes have been shown to be a risk factor of airway obstruction in cross-sectional studies in the general population. OBJECTIVES: Our aim was to study the relationships between specific occupations and occupational exposures during a 9-yr follow-up period and changes in lung function and symptoms of chronic bronchitis. METHODS: Subjects from the general population aged 20 to 45 yr were randomly selected in 1991-1993 within the European Community Respiratory Health Survey. Follow-up took place from 1998 to 2002 among 4,079 males and 4,461 females in 27 study centers. A total of 3,202 men and 3,279 women twice completed lung function measurements. Job history during follow-up was linked to a job exposure matrix and consequently translated into cumulative exposure estimates. MAIN RESULTS: Individuals exposed to dusts, gases, and fumes during the period of follow-up did not have a steeper decline of FEV1 than did individuals with consistently white-collar occupations without occupational exposures (relative change among men and women, + 1.4 and 3.1 ml/yr, respectively; p > 0.2), nor an increase of prevalence or incidence of airway obstruction defined as an FEV1/FVC ratio of less than 0.7. The incidence of chronic phlegm increased in men exposed to mineral dust (relative risk, 1.94 [1.29-2.91]) and gases and fumes (relative risk, 1.53 [0.99-2.36]), which was not modified by smoking. CONCLUSION: Occupational exposures to dusts, gases, and fumes occurring during the 1990s are associated with incidence of chronic bronchitis, although these did not impair lung function in a population of relatively young age. PMID- 16040785 TI - Heterogeneity of treatment response to azithromycin in patients with cystic fibrosis. AB - RATIONALE: We recently reported a randomized, placebo-controlled trial of azithromycin in patients with cystic fibrosis (CF) that demonstrated a 6.2% improvement in the 168-d relative change in FEV1 among azithromycin participants compared with placebo participants. OBJECTIVES: In the current analyses, heterogeneity of treatment response and the association between FEV1 and the risk of pulmonary exacerbations were investigated. METHODS: The time to first pulmonary exacerbation, hospitalization rates, and antibiotic use were compared between participants categorized by their relative change in FEV1 % predicted (>or= 5 vs. < 5% improvement) at Day 168. Pulmonary function and exacerbation responses were compared in subgroups of participants characterized by long-term concomitant medications and baseline lung function. MEASUREMENTS: All available data from the 185 randomized participants in the azithromycin trial were included in these analyses. MAIN RESULTS: Compared with placebo participants, a reduced risk of pulmonary exacerbations was observed both among azithromycin participants with >or= 5% and those with < 5% relative improvement in FEV1. Similarly, decreased hospitalization rates and decreased use of oral quinolone and nonquinolone antibiotics were observed in azithromycin participants regardless of improvement in FEV1. Subgroup analyses demonstrated that overall, participants on long-term aerosolized tobramycin and/or rhDNase had worse baseline lung function, but still benefited from azithromycin, as evidenced by a lower risk of exacerbations. CONCLUSIONS: Azithromycin participants experienced benefits in exacerbation parameters regardless of FEV1 response or subgroup. These data have implications for clinical practice and the design of clinical trials. PMID- 16040786 TI - Defective apoptotic cell phagocytosis attenuates prostaglandin E2 and 15 hydroxyeicosatetraenoic acid in severe asthma alveolar macrophages. AB - RATIONALE: Clearance of apoptotic cells is crucial to the resolution of inflammation and development of fibrosis, but the process is not well understood in normal or diseased human lungs. OBJECTIVES: To determine phagocytosis of apoptotic cells by primary human alveolar macrophages and whether defects in uptake of apoptotic cells are associated with decreases in antiinflammatory/antifibrotic mediators. METHODS: Human bronchoalveolar lavage macrophages (AMphis) from normal control subjects and subjects with mild-moderate or severe asthma were examined in vitro for phagocytosis of apoptotic human T cell line Jurkats and secretion of inflammatory mediators. MEASUREMENTS AND MAIN RESULTS: AMphis from normal subjects and patients with mild-moderate asthma were able to phagocytose apoptotic cells in response to LPS, resulting in an induction of the antifibrotic and/or antiinflammatory eicosanoids, prostaglandin E2 (PGE2) and 15-hydroxyeicosatetraenoic acid (HETE). In contrast, AMphis from patients with severe asthma had defective LPS-stimulated uptake of apoptotic cells, with associated failure to induce PGE2 and 15-HETE. In addition, LPS-stimulated basal levels of tumor necrosis factor alpha and granulocyte-macrophage colony stimulating factor were reduced in all patients with asthma, whereas PGE2 and 15 HETE were reduced only in patients with severe asthma. Dexamethasone enhanced specific uptake of apoptotic cells in all subjects, while suppressing inflammatory mediator secretion. CONCLUSIONS: A decrease in AMphis LPS responsiveness in severe asthma is manifested by defective apoptotic cell uptake and reduces secretion of inflammatory mediators. This may contribute to the chronicity of inflammation and remodeling in lungs of patients with asthma. PMID- 16040787 TI - Leukotrienes in acute lung injury: a potential therapeutic target? PMID- 16040788 TI - A urinary test for pulmonary arterial hypertension? PMID- 16040789 TI - Is it time to consider a new treatment for children with sleep-disordered breathing? PMID- 16040790 TI - Notes from the NHLBI Director: a vision for the future: opportunities and challenges. PMID- 16040791 TI - Curriculum and competency assessment tools for sleep disorders in pulmonary fellowship training programs. PMID- 16040793 TI - Anorexia nervosa and emphysema. PMID- 16040795 TI - Testosterone Supplementation during Respiratory Rehabilitation. PMID- 16040796 TI - Endoscopic ultrasound staging of lung cancer. PMID- 16040798 TI - Ground-based and remotely sensed nutrient availability across a tropical landscape. AB - Tropical soils often are assumed to be highly weathered and thus nutrient depleted, but this prediction applies primarily to geomorphically stable surfaces. Topography complicates the assumption of nutrient depletion, because erosion can enhance the supply of nutrients to tropical ecosystems. Consequently, understanding nutrient availability across landscapes requires a spatially explicit assessment of the relative strength of depletion and enhancement. We document the relationship between foliar nutrients and topographic position across a 20-km(2), 4- to 5-million-year-old eroded landscape in Kaua'i, Hawai'i, and use this relationship to build a bottom-up map of predicted nutrient availability across this landscape. Only approximately 17% of the landscape is nutrient-poor, mostly on stable uplands; nutrient availability on slopes and valley bottoms is much higher, in some cases similar to the most fertile montane forests in the Hawaiian Islands. This pattern was corroborated by top-down remote sensing of area-integrated canopy phosphorus concentrations. PMID- 16040799 TI - Toward a live microbial microbicide for HIV: commensal bacteria secreting an HIV fusion inhibitor peptide. AB - Most HIV transmission occurs on the mucosal surfaces of the gastrointestinal and cervicovaginal tracts, both of which are normally coated by a biofilm of nonpathogenic commensal bacteria. We propose to genetically engineer such naturally occurring bacteria to protect against HIV infection by secreting antiviral peptides. Here we describe the development and characterization of Nissle 1917, a highly colonizing probiotic strain of Escherichia coli, secreting HIV-gp41-hemolysin A hybrid peptides that block HIV fusion and entry into target cells. By using an appropriate combination of cis- and transacting secretory and regulatory signals, micromolar secretion levels of the anti-HIV peptides were achieved. The genetically engineered Nissle 1917 were capable of colonizing mice for periods of weeks to months, predominantly in the colon and cecum, with lower concentrations of bacteria present in the rectum, vagina, and small intestine. Histological and immunocytochemical examination of the colon revealed bacterial growth and peptide secretion throughout the luminal mucosa and in association with epithelial surfaces. The use of genetically engineered live microbes as anti HIV microbicides has important potential advantages in economy, efficacy, and durability. PMID- 16040800 TI - Chemical exchange 2D IR of hydrogen-bond making and breaking. AB - The involvement of chemical exchange in 2D IR heterodyne echo spectroscopy is characterized through the hydrogen-bond exchange between CH3OH and the CN of CH3CN. The exchange dynamics on the hydrogen-bond potential surfaces associated with different quantum states of the high-frequency CN stretching mode contributes to strong cross peaks between CN groups in two different chemical configurations and provides firm evidence of the hydrogen exchange between them. In analogy with NMR, the chemical exchange is seen in both slow and dynamic regimes. The relative magnitudes of the cross peaks at various population periods measure the picosecond regime time constants for H-bond transfer, whereas the temperature dependences indicate that the activation energy for the exchange from the H-bonded state to the free state is approximately 6.2 kJ.mol(-1). The results suggest that the hydrogen-bond dynamics is very similar in both vibrational quantum states of CN, suggesting that this stretching mode is not strongly coupled to the H-bond breaking reaction coordinate. The likely manifestations of chemical exchange in 2D IR experiments are discussed. PMID- 16040801 TI - The RNaseIII enzyme Dicer is required for morphogenesis but not patterning of the vertebrate limb. AB - The RNaseIII-containing enzyme Dicer is believed to be required for the processing of most, if not all, microRNAs (miRNAs) and for processing long dsRNA into small interfering RNAs. Because the complete loss of Dicer in both zebrafish and mice results in early embryonic lethality, it has been impossible to determine what role, if any, Dicer has in patterning later tissues in the developing vertebrate embryo. To bypass the early requirement of Dicer in development, we have created a conditional allele of this gene in mice. Using transgenes to drive Cre expression in discrete regions of the limb mesoderm, we find that removal of Dicer results in the loss of processed miRNAs. Phenotypically, developmental delays, in part due to massive cell death as well as disregulation of specific gene expression, lead to the formation of a much smaller limb. Thus, Dicer is required for the formation of normal mouse limbs. Strikingly, however, we did not detect defects in basic patterning or in tissue specific differentiation of Dicer-deficient limb buds. PMID- 16040802 TI - Chromatin immunoprecipitation cloning reveals rapid evolutionary patterns of centromeric DNA in Oryza species. AB - The functional centromeres of rice (Oryza sativa, AA genome) chromosomes contain two key DNA components: the CRR centromeric retrotransposons and a 155-bp satellite repeat, CentO. However, several wild Oryza species lack the CentO repeat. We developed a chromatin immunoprecipitation-based technique to clone DNA fragments derived from chromatin containing the centromeric histone H3 variant CenH3. Chromatin immunoprecipitation cloning was carried out in the CentO-less species Oryza rhizomatis (CC genome) and Oryza brachyantha (FF genome). Three previously uncharacterized genome-specific satellite repeats, CentO-C1, CentO-C2, and CentO-F, were discovered in the centromeres of these two species. An 80-bp DNA region was found to be conserved in CentO-C1, CentO, and centromeric satellite repeats from maize and pearl millet, species which diverged from rice many millions of years ago. In contrast, the CentO-F repeat shows no sequence similarity to other centromeric repeats but has almost completely replaced other centromeric sequences in O. brachyantha, including the CRR-related sequences that normally constitute a significant fraction of the centromeric DNA in grass species. PMID- 16040803 TI - Retrograde movement of tRNAs from the cytoplasm to the nucleus in Saccharomyces cerevisiae. AB - In eukaryotes, tRNAs transcribed in the nucleus function in cytoplasmic protein synthesis. The Ran-GTP-binding exportin, Los1p/Xpo-t, and additional pathway(s) mediate tRNA transport to the cytoplasm. Although tRNA movement was thought to be unidirectional, recent reports that yeast precursor tRNA splicing occurs in the cytoplasm, whereas fully spliced tRNAs can reside in the nucleus, require that either the precursor tRNA splicing machinery or mature tRNAs move from the cytoplasm to the nucleus. Our data argue against the first possibility and strongly support the second. Combining heterokaryon analysis with fluorescence in situ hybridization, we show that a foreign tRNA encoded by one nucleus can move from the cytoplasm to a second nucleus that does not encode the tRNA. We also discovered nuclear accumulation of endogenous cytoplasmic tRNAs in haploid yeast cells in response to nutritional deprivation. Nuclear accumulation of cytoplasmic tRNA requires Ran and the Mtr10/Kap111 member of the importin-beta family. Retrograde tRNA nuclear import may provide a novel mechanism to regulate gene expression in eukaryotes. PMID- 16040804 TI - Ganglioside GD3 promotes cell growth and invasion through p130Cas and paxillin in malignant melanoma cells. AB - Although ganglioside GD3 levels are highly elevated in malignant melanomas, the role of GD3 in melanomas' malignant properties has not been clearly shown. To investigate this problem, we genetically generated GD3-positive (GD3+) transfectant cells from a GD3-negative (GD3-) mutant line SK-MEL-28-N1 and analyzed the phenotypic changes in the transfected cells. GD3+ cells showed markedly increased cell growth and invasive characteristics. Two bands that underwent stronger tyrosine phosphorylation in GD3+ cell lines than in controls after treatment with FCS were found with molecular masses of 130 and 68 kDa. They were identified as p130Cas and paxillin by sequential immunoprecipitation. Their roles in cell growth and invasion were analyzed with a small interfering RNA (siRNA) approach. Cell growth, as analyzed by BrdUrd uptake, was strongly suppressed in GD3+ cells to near the levels of GD3- cells when treated with siRNA for p130Cas but not when treated with siRNA for paxillin. However, treatment with siRNAs of either p130Cas or paxillin resulted in the marked suppression of the invasive activity of GD3+ cells almost to the levels of control cells. These results suggested that these two molecules function as effectors of GD3-mediated signaling, leading to such malignant properties as rapid cell growth and invasion. PMID- 16040806 TI - Role of pro-IGF-II processing by proprotein convertase 4 in human placental development. AB - Fetal growth restriction (intrauterine growth restriction, IUGR) is a leading cause of perinatal mortality. However, the causes of aberrant development of the placenta and, thus, of the fetus, are not currently known. Insulin-like growth factor II (IGF-II) has been shown to be an important regulator of fetoplacental growth. This growth factor must undergo posttranslational processing, and, thus, we hypothesized that aberrant processing of pro-IGF-II to IGF-II may be a cause of IUGR. Here, we have found that the proprotein convertase PC4 is expressed in the human placenta and that it cleaves pro-IGF-II to generate the intermediate processed form, IGF-II (1-102) and, subsequently, mature IGF-II (1-67), which are accounted for by the removal of terminal basic residues by carboxypeptidases. This processing confers the ability of IGF-II to activate invasive trophoblast cells through AKT phosphorylation, whereas inhibition of PC4 by a PC4-specific inhibitor blocks pro-IGF-II processing and reduces trophoblast cell migration, which can be partly restored by addition of mature IGF-II. Consistent with the hypothesis that IGF-II processing is implicated in IUGR, sera of patients carrying IUGR fetuses displayed elevated levels of pro-IGF-II. Thus, abnormal processing of IGF-II by PC4 may represent a previously uncharacterized mechanism involved in the pathophysiology of fetoplacental growth restriction, and elevated pro-IGF-II may be a useful clinical marker for risk of IUGR. PMID- 16040805 TI - Docosahexaenoic acid: a positive modulator of Akt signaling in neuronal survival. AB - Phosphatidylinositol 3-kinase [PI (3)K]/Akt signaling is a critical pathway in cell survival. Here, we demonstrate a mechanism where membrane alteration by the n-3 fatty acid status affects Akt signaling, impacting neuronal survival. Docosahexaenoic acid (DHA), an n-3 polyunsaturated fatty acid highly enriched in neuronal membranes, promotes neuronal survival by facilitating membrane translocation/activation of Akt through its capacity to increase phosphatidylserine (PS), the major acidic phospholipid in cell membranes. The activation of PI (3)K and phosphatidylsinositol triphosphate formation were not affected by DHA, indicating that membrane interaction of Akt is the event responsible for the DHA effect. Docosapentaenoic acid, which replaces DHA during n-3 fatty acid deficiency, was less effective in accumulating PS and translocating Akt and thus less effective in preventing apoptosis. Consistently, in vivo reduction of DHA by dietary depletion of n-3 fatty acids decreased hippocampal PS and increased neuronal susceptibility to apoptosis in cultures. This mechanism may contribute to neurological deficits associated with n-3 fatty acid deficiency and support protective effects of DHA in pathological models such as brain ischemia or Alzheimer's disease. PMID- 16040807 TI - DNA-based vaccines activate innate and adaptive antitumor immunity by engaging the NKG2D receptor. AB - The interaction of NKG2D, a stimulatory receptor expressed on natural killer (NK) cells and activated CD8(+) T cells, and its ligands mediates stimulatory and costimulatory signals to these cells. Here, we demonstrate that DNA-based vaccines, encoding syngeneic or allogeneic NKG2D ligands together with tumor antigens such as survivin or carcinoembryonic antigen, markedly activate both innate and adaptive antitumor immunity. Such vaccines result in highly effective, NK- and CD8(+) T cell-mediated protection against either breast or colon carcinoma cells in prophylactic and therapeutic settings. Notably, this protection was irrespective of the NKG2D ligand expression level of the tumor cells. Hence, this strategy has the potential to lead to widely applicable and possibly clinically useful DNA-based cancer vaccines. PMID- 16040808 TI - Expression, physiological action, and coexpression patterns of neuropeptide Y in rat taste-bud cells. AB - Recent studies have suggested that neuropeptides could play previously unrecognized functional roles in peripheral gustation. To date, two peptides, cholecystokinin and vasoactive intestinal peptide, have been localized to subsets of taste-bud (TB) cells (TBC) and one, cholecystokinin, has been demonstrated to produce excitatory physiological actions. This study extends our knowledge of neuropeptides in TBC in three significant ways. First, using techniques of immunocytochemistry and RT-PCR, evidence is presented for the expression of a third peptide, neuropeptide Y (NPY). Like other peptide expression patterns, NPY expression is circumscribed to a subset of cells within the taste bud. Second, using physiological studies, we demonstrate that NPY specifically enhances an inwardly rectifying potassium current via NPY-Y1 receptors. This action is antagonistic to the previously demonstrated inhibitory effect exerted by cholecystokinin on the same current, thus providing important clues to their signaling roles in the TB. Third, using the technique of double-labeled fluorescent immunocytochemistry, the relationship of three subsets of neuropeptide-expressing TB cells to one another was examined. Remarkably, NPY expressions, although fewer in number than either the cholecystokinin or vasoactive intestinal peptide subsets, overlapped 100% with either peptide. Collectively, these three observations transform previously suggestive roles of neuromodulation by peptides in TB cells to more concrete signaling pathways. The extensive colocalization of these peptides suggests they may be subject to similar presynaptic influences of release yet have antagonistic postsynaptic actions. The convergence or divergence of these postsynaptic actions awaits further investigation. PMID- 16040809 TI - The high-pressure phase of alumina and implications for Earth's D'' layer. AB - Using ab initio simulations and high-pressure experiments in a diamond anvil cell, we show that alumina (Al(2)O(3)) adopts the CaIrO(3)-type structure above 130 GPa. This finding substantially changes the picture of high-pressure behavior of alumina; in particular, we find that perovskite structure is never stable for Al(2)O(3) at zero Kelvin. The CaIrO(3)-type phase suggests a reinterpretation of previous shock-wave experiments and has important implications for the use of alumina as a window material in shock-wave experiments. In particular, the conditions of the stability of this phase correspond to those at which shock-wave experiments indicated an increase of the electrical conductivity. If this increase is caused by high ionic mobility in the CaIrO(3)-type phase of Al(2)O(3), similar effect can be expected in the isostructural postperovskite phase of MgSiO(3) (which is the dominant mineral phase in the Earth's D'' layer). The effect of the incorporation of Al on the perovskite/postperovskite transition of MgSiO(3) is discussed. PMID- 16040810 TI - Constitutively active L-type Ca2+ channels. AB - Ca(2+) influx through L-type Ca(2+) channels (LTCCs) influences numerous physiological processes ranging from contraction in muscle and memory in neurons to gene expression in many cell types. However, the spatiotemporal organization of functional LTCCs has been nearly impossible to investigate because of methodological limitations. Here, we examined LTCC function with high temporal and spatial resolution using evanescent field fluorescence microscopy. Surprisingly, we found that LTCCs operated in functionally organized clusters, not necessarily as individual proteins. Furthermore, LTCC function in these clusters does not appear to be controlled by simple stochastic gating but instead by a PKC-dependent switch mechanism. This work suggests that resting intracellular free calcium concentration in arterial myocytes is predominantly controlled by this process in combination with rare voltage-dependent openings of individual LTCCs. We propose that Ca(2+) influx via persistent LTCCs may be an important mechanism regulating steady-state local and global Ca(2+) signals. PMID- 16040811 TI - Giardia mitosomes and trichomonad hydrogenosomes share a common mode of protein targeting. AB - Mitochondria are archetypal organelles of endosymbiotic origin in eukaryotic cells. Some unicellular eukaryotes (protists) were considered to be primarily amitochondrial organisms that diverged from the eukaryotic lineage before the acquisition of the premitochondrial endosymbiont, but their amitochondrial status was recently challenged by the discovery of mitochondria-like double membrane bound organelles called mitosomes. Here, we report that proteins targeted into mitosomes of Giardia intestinalis have targeting signals necessary and sufficient to be recognized by the mitosomal protein import machinery. Expression of these mitosomal proteins in Trichomonas vaginalis results in targeting to hydrogenosomes, a hydrogen-producing form of mitochondria. We identify, in Giardia and Trichomonas, proteins related to the component of the translocase in the inner membrane from mitochondria and the processing peptidase. A shared mode of protein targeting supports the hypothesis that mitosomes, hydrogenosomes, and mitochondria represent different forms of the same fundamental organelle having evolved under distinct selection pressures. PMID- 16040812 TI - Formation of morphologically similar globular aggregates from diverse aggregation prone proteins in mammalian cells. AB - Huntington's disease is a progressive neurodegenerative disorder caused by a polyglutamine repeat expansion in the first exon of the huntingtin (Htt) protein. N-terminal Htt peptides with polyglutamine tracts in the pathological range (51 122 glutamines) form high-molecular-weight protein aggregates with fibrillar morphology in vitro, and they form discrete inclusion bodies in a cell-culture model. However, in some studies, formation of discrete Htt inclusions does not correlate well with cell death. We coexpressed N-terminal Htt fragments containing 91 glutamines fused to different affinity tags in HEK293 cells, and we isolated small aggregates by double sequential-affinity chromatography to assure the isolation of multimeric molecules. Transmission electron microscopy and atomic force microscopy revealed the isolated aggregates as globules or clusters of globules 4-50 nm in diameter without any detectable fibrillar species. Because small nonfibrillar oligomers, not mature fibrils, recently have been suggested to be the principal cytotoxic species in neurodegenerative disease, these Htt globular aggregates formed in cells may represent the pathogenic form of mutant Htt. PMID- 16040813 TI - Nicotine regulates DARPP-32 (dopamine- and cAMP-regulated phosphoprotein of 32 kDa) phosphorylation at multiple sites in neostriatal neurons. AB - Nicotinic acetylcholine receptors (nAChRs) regulate dopaminergic signaling in the striatum by modulating the release of neurotransmitters. We have recently reported that nicotine stimulates the release of dopamine via alpha4beta2(*) nAChRs and/or alpha7 nAChRs, leading to the regulation of DARPP-32 at Thr34, the site involved in regulation of protein phosphatase-1 (PP-1). In this study, we investigated the regulation of DARPP-32 phosphorylation at its other sites, Thr75 [cyclin-dependent kinase-5 (Cdk5) site], Ser97 (CK2 site), and Ser130 (CK1 site), that serve to modulate Thr34 phosphorylation and dephosphorylation. In neostriatal slices, nicotine (100 microM) increased phosphorylation of DARPP-32 at Ser97 and Ser130 at an early time point (30 s) and decreased phosphorylation of DARPP-32 at Thr75 at a late time point (3 min). The increase in Ser97 and Ser130 phosphorylation was mediated through the release of dopamine via activation of alpha4beta2(*) nAChRs and alpha7 nAChRs and the subsequent activation of dopamine D1 and D2 receptors. The decrease in Thr75 phosphorylation was mediated through the release of dopamine via activation of alpha4beta2(*) nAChRs and the subsequent activation of dopamine D1 receptors. These various actions of nicotine on modulatory sites of phosphorylation would be predicted to result in a synergistic increase in the state of phosphorylation of DARPP-32 at Thr34 and thus would contribute to increased dopamine D1 receptor/DARPP-32 Thr34/PP-1 signaling. PMID- 16040814 TI - Fenobam: a clinically validated nonbenzodiazepine anxiolytic is a potent, selective, and noncompetitive mGlu5 receptor antagonist with inverse agonist activity. AB - Fenobam [N-(3-chlorophenyl)-N'-(4,5-dihydro-1-methyl-4-oxo-1H-imidazole-2 yl)urea] is an atypical anxiolytic agent with unknown molecular target that has previously been demonstrated both in rodents and human to exert anxiolytic activity. Here, we report that fenobam is a selective and potent metabotropic glutamate (mGlu)5 receptor antagonist acting at an allosteric modulatory site shared with 2-methyl-6-phenylethynyl-pyridine (MPEP), the protypical selective mGlu5 receptor antagonist. Fenobam inhibited quisqualate-evoked intracellular calcium response mediated by human mGlu5 receptor with IC(50) = 58 +/- 2 nM. It acted in a noncompetitive manner, similar to MPEP and demonstrated inverse agonist properties, blocking 66% of the mGlu5 receptor basal activity (in an over expressed cell line) with an IC(50) = 84 +/- 13 nM. [(3)H]Fenobam bound to rat and human recombinant receptors with K(d) values of 54 +/- 6 and 31 +/- 4 nM, respectively. MPEP inhibited [(3)H]fenobam binding to human mGlu5 receptors with a K(i) value of 6.7 +/- 0.7 nM, indicating a common binding site shared by both allosteric antagonists. Fenobam exhibits anxiolytic activity in the stress induced hyperthermia model, Vogel conflict test, Geller-Seifter conflict test, and conditioned emotional response with a minimum effective dose of 10 to 30 mg/kg p.o. Furthermore, fenobam is devoid of GABAergic activity, confirming previous reports that fenobam acts by a mechanism distinct from benzodiazepines. The non-GABAergic activity of fenobam, coupled with its robust anxiolytic activity and reported efficacy in human in a double blind placebo-controlled trial, supports the potential of developing mGlu5 receptor antagonists with an improved therapeutic window over benzodiazepines as novel anxiolytic agents. PMID- 16040815 TI - Coformulated N-octanoyl-glucosylceramide improves cellular delivery and cytotoxicity of liposomal doxorubicin. AB - The anticancer agent doxorubicin is in certain cases administered as a long circulating liposomal formulation. Due to angiogenesis-related structural abnormalities in the endothelial lining of many neoplasms, these complexes tend to extravasate and accumulate in the tumor stroma. However, delivery of doxorubicin is still not optimal since liposomes are not taken up directly by tumor cells. Instead, doxorubicin is gradually released into the interstitial space, and the subsequent uptake by surrounding cells is a limiting step in the delivery process. We recently demonstrated that plasma membrane-inserted short chain sphingomyelin facilitates the cellular uptake of free doxorubicin. Here, we report that N-octanoyl-glucosylceramide acts equally potent but is itself less toxic. When coformulated with liposomal doxorubicin, this short-chain glycosphingolipid administered to cultured A431 epidermoid carcinoma cells led to superior (up to 4-fold) cellular doxorubicin accumulation and cytotoxicity, compared with control doxorubicin liposomes. These results were fully reproducible when N-octanoyl-glucosylceramide was postinserted into Caelyx, a commercial liposomal doxorubicin preparation. The doxorubicin-potentiating effect of N-octanoyl-glucosylceramide-enriched liposomes proved relatively insensitive to high serum concentrations, indicating that in vivo application is a feasible option. N-Octanoyl-glucosylceramide enrichment might thus represent a major improvement of conventional liposomal doxorubicin formulations. PMID- 16040816 TI - Ultrasonography in tarsal tunnel syndrome. AB - OBJECTIVE: The purpose of this study was to clarify the diagnostic value of ultrasonography in tarsal tunnel syndrome. METHODS: Seventeen patients (17 feet) with tarsal tunnel syndrome were treated between 1988 and 2003. Preoperative ultrasonography was performed, and the cause of the syndrome was confirmed intraoperatively in all cases. Long and short axes of the tarsal tunnel were scanned to ascertain the presence of any space-occupying lesion. RESULTS: The causes of tarsal tunnel syndrome, as confirmed by surgery, were ganglia (n = 10), talocalcaneal coalition (n = 1), talocalcaneal coalition associated with ganglia (n = 3), and varicose veins (n = 3). Among the cases involving ganglia, hypoechoic or anechoic regions were observed. The mean sizes +/- SD of these regions were 19.4 +/- 8.8 mm in the long axis, 15.2 +/- 6.3 mm in the short axis, and 10.4 +/- 3.8 mm in depth. Of these, 3 ganglia were not clearly palpable before surgery and were small: 10 x 10 x 7, 13 x 11 x 9, and 9 x 8 x 7 mm. Among the cases involving talocalcaneal coalition, ultrasonography indicated a beak shaped bony process on the short axis images. Although these 3 cases were associated with ganglia, this could not be determined by preoperative palpation. CONCLUSIONS: As a diagnostic imaging technique for tarsal tunnel syndrome, ultrasonography is extremely useful for identifying space-occupying lesions. Ultrasonography should be performed routinely in patients with suspected tarsal tunnel syndrome. PMID- 16040817 TI - Clinical utility of sonography in diagnosing plantar fasciitis. AB - OBJECTIVE: The purpose of this study was to investigate the efficacy of sonography in the detection of plantar fasciitis (PF) compared with magnetic resonance imaging (MRI) findings in subjects with inferior heel pain. METHODS: Seventy-seven patients with unilateral (n = 9) and bilateral (n = 68) heel pain were studied. Seventy-seven age- and sex-matched asymptomatic subjects served as a control group. Magnetic resonance imaging was used to establish a diagnosis of PF with sagittal T1-weighted, T2-weighted, and short tau inversion recovery sequences. The sonographic appearances of PF were compared with MRI findings. Plantar fascia and heel pad thickness were also measured on both imaging modalities. RESULTS: Compared with MRI, sonography showed 80% sensitivity and 88.5% specificity in assessing PF. A strong correlation was found between plantar fascia and fat pad thickness measurements done by sonography (P < .001; r = 0.854) and MRI (P < .001; r = 0.798). Compared with the asymptomatic volunteers, patients with PF had significant increases in plantar fascia and heel pad thicknesses, weight, and body mass index (P = .0001). Heel pad thickness was also significantly increased with pain duration (P = .021). CONCLUSIONS: Although MRI is the modality of choice in the morphologic assessment of different plantar fascia lesions, sonography can also serve as an effective tool and may substitute MRI in the diagnosis of PF. PMID- 16040818 TI - The clinical significance of an empty renal fossa on prenatal sonography. AB - OBJECTIVE: The purpose of this study was to characterize the significance of an empty renal fossa on prenatal sonography. METHODS: A hospital database was reviewed retrospectively to find all fetuses with an empty renal fossa diagnosed on prenatal sonography between 1989 and 2003. For each case, prenatal and postnatal data were recorded. RESULTS: Ninety-three fetuses had an empty renal fossa: 49 on the left, 41 on the right, and 3 unspecified. Forty-four fetuses were male and 29 were female. For 20, the sex was not recorded. The average gestational age at diagnosis was 29.5 weeks. Final diagnoses were made on the basis of prenatal and postnatal imaging. The missing kidney was ectopic in 39 cases (42%): 34 in the pelvis, 4 fused to the contralateral kidney, and 1 in the thorax due to a congenital diaphragmatic hernia. The absent kidney was never located in 44 cases (47%) and presumed to be congenitally absent. Ten kidneys (11%) originally thought absent were normally located, 7 of which were dysplastic, 2 normal, and 1 infiltrated by a tumor. In 39 patients (42%), other anomalies were identified, sometimes involving multiple systems, most commonly genitourinary (29) and cardiovascular (13). Of the 77 cases with the number of umbilical cord vessels recorded, 68 (88%) were normal and 9 (12%) had only 2 vessels. CONCLUSIONS: If a kidney is not found in the renal fossa, most are either ectopic (42%) or congenitally absent (47%). An empty renal fossa is often (42%) associated with other congenital anomalies. PMID- 16040819 TI - Does the frequency of soft sonographic aneuploidy markers vary by fetal sex? AB - OBJECTIVE: To determine whether the frequency of soft sonographic aneuploidy markers varies by fetal sex. METHODS: We identified all singleton fetuses with known sex undergoing genetic sonography at 17 weeks' to 21 weeks 6 days' gestation in a single perinatal center from January 1, 2000, to December 31, 2003. Markers studied were biparietal diameter/femur length, transcerebellar diameter, ear length, echogenic bowel, femur length, humerus length, absent middle fifth phalanx, nuchal fold, renal pelvis dilatation, echogenic cardiac focus, and choroid plexus cysts. Additional information extracted from the prospectively ascertained database included maternal age, referral indications, and chromosomal analyses. Multiple gestations and fetuses with structural or chromosomal abnormalities were excluded. The study received exempt review status by the Institutional Review Board. Dichotomous variables were compared by the chi(2) or Fisher exact test; continuous variables were compared by the unpaired t test. RESULTS: In total, 4057 eligible fetuses, 2103 male and 1954 female, were examined at 18.9 +/- 0.9 weeks (mean +/- SD). Referral indications included maternal age of 35 years or older (n = 2983), abnormal second-trimester serum screen results (n = 610), soft marker on sonography (n = 583), prior aneuploid offspring (n = 24), and other (n = 125). More than 1 referral indication was possible for a given fetus. Overall, male fetuses exhibited echogenic fetal bowel (odds ratio, 1.76; 95% confidence interval [CI], 1.14-2.72; P = .009) and renal pelvis dilatation (odds ratio, 2.00; 95% CI, 1.30-3.09; P = .001) significantly more often than female fetuses. However, when fetuses were evaluated for single isolated markers, only male predominance of renal pelvis dilatation persisted (odds ratio, 2.32; 95% CI, 1.32-4.09; P = .003). No markers had increased frequency in female offspring. CONCLUSIONS: Male fetuses exhibit a significantly increased frequency of renal pelvis dilatation compared with female fetuses. Sex specific adjustment of sonographically derived aneuploidy risk does not appear to be indicated. However, a larger series of fetuses with trisomy 21 and pyelectasis is required to assess sex-specific risk adjustment for this marker. PMID- 16040820 TI - Differentiation of vascular dementia and Alzheimer disease: a functional transcranial Doppler ultrasonographic study. AB - OBJECTIVE: In brain perfusion studies, perfusion defects have been reported mainly in the temporal and parietal regions in patients with Alzheimer disease (AD), but it is known that the occipital cortex is partially preserved from metabolic defects in patients with AD, at least in the early stage of the disease. We therefore evaluated the reactivity of the posterior cerebral arteries during visual stimulation with transcranial Doppler ultrasonography (TDU). METHODS: Fifteen patients with AD (mean age, 70 years), 12 patients with vascular dementia (VD) (mean age, 61.5 years), and 9 healthy control subjects (mean age, 58 years) were enrolled in the study. The reactivity of both posterior cerebral arteries during visual stimulation was measured with TDU. Reactivity was calculated by the differences in mean relative blood flow velocity (rBFv) between stimulation (vs) and rest (vr) divided by the resting value [rBFv = 100 x (vs - vr)/vr]. RESULTS: Significant increases of vascular reactivity were obtained during visual stimulation for each group (P < .001). There was no significant right or left side difference for vascular reactivity within the groups except in VD; therefore, values of the right and left sides were pooled for patients with AD and control subjects. Reactivity to visual stimulation was similar for the patients with AD (mean +/- SD, 38% +/- 2.2%) and control subjects (35% +/- 1.8%), but reactivity was found significantly decreased in both sides of patients with VD (25.7% +/- 2.1% for the left side and 24% +/- 2.6% for the right side) compared with the healthy control subjects and the patients with AD (P < .001). CONCLUSIONS: These results indicate that the occipital cortex is partially preserved in patients with AD and that functional TDU may be a valuable tool for differentiating VD and AD. PMID- 16040821 TI - Detection of cardiac right-to-left shunts by contrast-enhanced harmonic carotid duplex sonography. AB - OBJECTIVE: Paradoxical embolization by cardiac right-to-left shunts (RLS) is increasingly recognized as an important factor for embolic stroke. Contrast enhanced transcranial Doppler sonography (ce-TCDS) is an established diagnostic tool for RLS detection but is frequently limited because of an inadequate temporal acoustic bone window. The purpose of this study was to determine whether extracranial sonography (ECS) using harmonic frequencies improves detection of RLS. METHODS: Extracranial color duplex sonography using harmonic frequencies enables visualization of even single ultrasound contrast agent microbubbles because of oscillation. Patients with stroke and positive RLS findings on transesophageal echocardiography underwent a simultaneous extracranial and transcranial sonographic examination of the proximal common carotid artery (CCA) and middle cerebral artery (MCA) on the same side. A Valsalva strain was performed for 10 seconds after intravenous bolus injection of a galactose-based nontranspulmonary contrast agent. The B-mode frame sequences of the transverse plane of the CCA obtained by harmonic ECS and the ce-TCDS recordings of high intensity transient signals from the MCA were analyzed offline. RESULTS: In all patients with RLS, the shunts could be identified by harmonic ECS. A close correlation could be seen between the count of visualized microbubbles in the CCA and the number of high-intensity transient signals detected on ce-TCDS in the ipsilateral MCA. CONCLUSIONS: The results of this study indicate that contrast enhanced ultrasound harmonic imaging of the CCA using a Valsalva strain might be an optional screening tool for detection of cardiac RLS in patients with insufficient acoustic bone windows. PMID- 16040822 TI - Ruptured abdominal aortic aneurysm: categorization of sonographic findings and report of 3 new signs. AB - OBJECTIVE: The purpose of this study was to categorize the sonographic findings seen in patients with a ruptured abdominal aortic aneurysm (AAA) and to describe 3 previously undescribed sonographic findings. METHODS: From January 1997 to December 2003, we evaluated 388 consecutive patients with an AAA (transverse aortic diameter >30 mm). Among these patients, 29 had surgical or computed tomographic demonstration of aneurysm rupture. The remaining 359 were asymptomatic and had no evidence of AAA rupture at follow-up. RESULTS: Findings recognized among 29 positive cases included AAA deformation (n = 12), luminal thrombus inhomogeneity (n = 20), clear interruption of a luminal thrombus (n = 5), retroperitoneal hematoma (n = 22), and hemoperitoneum (n = 11). In addition, 3 previously unreported findings were noted: an intraluminal floating thrombus layer (n = 8), a parietal hypoechoic focus due to aneurysm wall interruption (n = 3), and a para-aortic hypoechoic area adjacent to the bleeding side (n = 4). Aside from AAA deformation and thrombus heterogeneity, no other signs were recognized among subjects with a nonruptured aneurysm. CONCLUSIONS: In the past, sonography has been used mainly to rapidly confirm aneurysm presence in the clinical setting of a patient with a suspected rupture. Instead, this retrospective series shows how this imaging technique can frequently identify several direct and indirect signs of aneurysm rupture itself. Moreover, 3 new indicators of AAA rupture have been observed. PMID- 16040823 TI - Venous insufficiency in patients with toenail onychomycosis. AB - OBJECTIVE: Onychomycosis is a common fungal infection of the toenails and can originate secondary to vascular abnormalities. The aim of this study was to evaluate the relationship between onychomycosis and venous insufficiency. METHODS: Forty-two patients with onychomycosis and 39 healthy control subjects who had normal toenails were enrolled in the study. Doppler examinations were performed with a commercially available scanner and a 7.5-MHz linear probe. Major superficial and deep veins of the lower limb, including long and short saphenous, femoral, and popliteal veins, were examined. Venous insufficiency was assessed with the Valsalva test. With the Doppler examination, retrograde flow of more than 1 second was accepted as venous insufficiency. RESULTS: Venous insufficiency was detected more frequently in patients with onychomycosis than in the control group (15 [35.7%] of 42 and 6 [15.4%] of 39, respectively; P = .037). Reflux was bilateral in 4 (26.7%) of 15 patients with onychomycosis, and in those 4 patients the onychomycosis was also bilateral. In 7 (46.7%) of 15 patients, onychomycosis and venous insufficiency were detected ipsilaterally, whereas there were no onychopathic features contralaterally. Although unilateral insufficiency was present in 4 (26.7%) of 15 patients, these patients had bilateral onychomycosis. CONCLUSIONS: We found a significant relationship between onychomycosis and venous insufficiency; therefore, we recommend a routine venous Doppler examination for patients with onychomycosis to diagnose or rule out venous insufficiency. PMID- 16040824 TI - Endometrial volume and vascularity measurements by transvaginal 3-dimensional ultrasonography and power Doppler angiography in stimulated and tumoral endometria: an interobserver reproducibility study. AB - OBJECTIVE: The purpose of this study was to evaluate interobserver reproducibility of endometrial volume and vascular indices of the endometrium and subendometrial area estimated by 3-dimensional power Doppler angiography (3D-PDA) using the Virtual Organ Computer-Aided Analysis program, determining the influence of the endometrial growth etiology on measurements. METHODS: Forty women underwent 3D-PDA ultrasonography. Group A comprised 25 women scanned on the day after controlled ovarian stimulation with human chorionic gonadotropin. Group B comprised 15 patients who had uterine bleeding and questionable endometrial thickening. (Histologic evaluation revealed 10 endometrial cancers and 5 endometrial hyperplasias.) A single observer examined all patients and acquired all volume data sets. Forty volume data sets were then analyzed with the Virtual Organ Computer-Aided Analysis program by 2 different observers. Endometrial volume and vascularity indices (vascularization index [VI], flow index [FI], and vascularization flow index [VFI]) of the endometrium and subendometrium were manually calculated in the coronal plane with a 9 degrees rotation step. An intraclass correlation coefficient (ICC) was used to assess interobserver reliability. RESULTS: Endometrial volume was more reproducible in group A (ICC = 0.98) than in group B (ICC = 0.58) (P < .05). Endometrial and subendometrial VI, FI, and VFI also presented good reproducibility with ICC greater than 0.84. The ICC was not statistically different for endometrial and subendometrial VI, FI, and VFI according to patient group, although subendometrial VFI was less reproducible in group B (ICC = 0.53) than in group A (ICC = 0.88). CONCLUSIONS: Endometrial volume and endometrial and subendometrial 3D power Doppler indices have acceptable reproducibility. The interobserver reproducibility in tumoral endometrium was more similar than in stimulated endometrium. Our results indicate that 3D-PDA is a reliable method to evaluate physiologic and pathologic endometrial changes. PMID- 16040825 TI - Incidence of cardiac arrhythmias with therapeutic versus diagnostic ultrasound and intravenous microbubbles. AB - OBJECTIVE: The purpose of this study was to determine the type of arrhythmias induced with therapeutic versus diagnostic transthoracic low-frequency ultrasound (TLFUS) transducers in the presence of intravenous microbubbles. METHODS: Intravenous perfluorocarbon-exposed sonicated dextrose albumin (PESDA) microbubbles were infused or given as a bolus injection while TLFUS was applied in the standard parasternal and apical views with either a 1-MHz therapeutic ultrasound transducer or high-mechanical-index diagnostic ultrasound (1.7 MHz). RESULTS: Significantly more ectopy was produced by the therapeutic transducer, especially at higher-intensity settings in the continuous wave mode after bolus injections of PESDA (P < .001 compared with lower intensities and lower continuous infusion rates). Six patients (15%) had either clinical supraventricular tachycardia or nonsustained ventricular tachycardia after intravenous PESDA with therapeutic TLFUS. In comparison, diagnostic high mechanical-index ultrasound produced only isolated ventricular ectopy and no sustained ventricular arrhythmias. CONCLUSIONS: Intravenously injected microbubbles and low-frequency therapeutic transducers operating at longer duty cycles and wide beam widths have the capability of eliciting clinically important arrhythmias in patients at high risk for such events. PMID- 16040826 TI - Accuracy of 3-dimensional color Doppler-derived flow volumes with increasing image depth. AB - OBJECTIVES: We and others have reported on the use of digital color Doppler sonography from real-time 3-dimensional (3D) echocardiography and its use in accurately calculating cardiac flow volumes, namely stroke volume (SV) and, hence, cardiac output. However, in some patients, image depth is higher than average, and this may affect the accuracy of volume calculation. We sought to investigate the impact of image depth and the accompanying change in signal strength, spatial resolution, and pulse repetition frequency on the accuracy of SV calculation from 3D color Doppler data in an in vitro model. METHODS: A tube model of the left ventricular outflow tract was constructed from plastic tubing and connected to a pulsatile pump. The volume flowing through the tube was imaged using a 3D echocardiography system. Stroke volumes from the pump were computed from the DICOM data using commercially available software and compared with a reference standard of timed volumes with the use of a graduated measuring cylinder over a range of depth settings and SVs. RESULTS: There was good correlation between the 3D-derived SVs and the reference cylinder measures over all depths from 4 to 16 cm at 1-cm increments with a tube diameter of 17 mm, a pump rate of 60 beats/min, and SVs ranging from 20 to 70 mL. The average r(2) value for the 13 different depths was 0.976. However, the accuracy of the 3D method of volume calculation appeared to fall at depths greater than 13 cm, especially at higher SVs. CONCLUSIONS: Stroke volume calculation from real-time 3D color Doppler data in this in vitro study shows that at depths greater than approximately 13 cm, accuracy decreases, especially at higher SVs. This may be due to decreased resolution and the reduced frame rate at these depths. At shallower depths, volume calculation form the 3D Doppler data appears very accurate. PMID- 16040827 TI - A magnetic resonance imaging-compatible, large-scale array for trans-skull ultrasound surgery and therapy. AB - OBJECTIVE: Advances in ultrasound transducer array and amplifier technologies have prompted many intriguing scientific proposals for ultrasound therapy. These include both mildly invasive and noninvasive techniques to be used in ultrasound brain surgery through the skull. In previous work, it was shown how a 500-element hemisphere-shaped transducer could correct the wave distortion caused by the skull with a transducer that operates at a frequency near 0.8 MHz. Because the objective for trans-skull focusing is its ultimate use in a clinical context, a new hemispheric phased-array system has now been developed with acoustic parameters that are optimized to match the values determined in preliminary studies. METHODS: The transducer was tested by focusing ultrasound through ex vivo human skulls and into a brain phantom by means of a phase-adaptive focusing technique. Simultaneously, the procedure was monitored by the use of magnetic resonance guidance and thermometry. RESULTS: The ultrasound focus of a 500 element 30-cm-diameter, 0.81-MHz array could be steered electronically through the skull over a volume of approximately 30 x 30 x 26 mm. Furthermore, temperature monitoring of the inner and outer surfaces of the skull showed that the array could coagulate targeted brain tissue without causing excessive skull heating. CONCLUSIONS: The successful outcome of these experiments indicates that intensities high enough to destroy brain tissue can be produced without excessive heating of the surrounding areas and without producing large magnetic resonance noise and artifacts. PMID- 16040828 TI - Diagnosing carotid stenosis by Doppler sonography: state of the art. AB - OBJECTIVE: The goal of this review article is to present the state of the art in the clinical applications and technical performance and interpretation of carotid sonographic examinations. METHODS: Relevant publications regarding color and duplex Doppler sonography (CDDS) of the carotid arteries extracted from a computerized database (MEDLINE) and from references cited in these articles not appearing on the Internet were reviewed. RESULTS: The ability to quickly and efficiently identify stenosis in the carotid artery is an important goal for clinicians and vascular surgeons. Identification of potentially treatable carotid stenosis enables selection of appropriate candidates for endarterectomy or stent implantation. Advances in performance and interpretation of carotid sonographic studies over the last 20 years have been driven by technological improvements in gray scale and CDDS examinations and have made carotid sonography an important means to reach this goal. On the basis of CDDS, intima-media thickness measurements and plaque location and characterization on gray scale imaging, flow disturbance and areas of stenosis on color Doppler sonography, and flow velocities on spectral Doppler sonography are obtained. The degree of the diameter of a stenosis of the internal carotid artery is the main parameter used for therapeutic approaches. Advantages and limitations of the method are included. CONCLUSIONS: Carotid sonography is a unique imaging method for the investigation of carotid abnormalities. Noninvasive, accurate, and cost effective, it provides morphologic and functional information. It is increasingly becoming the first and often the sole imaging study before endarterectomy, whereas costly and invasive procedures are reserved for special cases. PMID- 16040829 TI - Rescue collateral flow in color duplex sonography. PMID- 16040830 TI - Transesophageal echocardiographic recognition of mitral valve aneurysm. PMID- 16040831 TI - Ultrasonographic evaluation of a vascularized tracheal transplantation. PMID- 16040832 TI - High-resolution ultrasonography in an aggressive thenar intramuscular lipoma. PMID- 16040833 TI - Ultrasonographic manifestations of fallopian tube carcinoma in the fimbriated end. PMID- 16040834 TI - Accessory pseudophallus with accessory pseudoscrotum detected during antenatal sonographic scanning. PMID- 16040835 TI - Prediction of methotrexate treatment failure in ectopic pregnancy. PMID- 16040836 TI - A critical role of a carboxylate in proton conduction by the ATP-binding cassette multidrug transporter LmrA. AB - The ATP binding cassette (ABC) transporter LmrA from the bacterium Lactococcus lactis is a homolog of the human multidrug resistance P-glycoprotein (ABCB1), the activity of which impairs the efficacy of chemotherapy. In a previous study, LmrA was shown to mediate ethidium efflux by an ATP-dependent proton-ethidium symport reaction in which the carboxylate E314 is critical. The functional importance of this key residue for ABC proteins was suggested by its conservation in a wider family of related transporters; however, the structural basis of its role was not apparent. Here, we have used homology modeling to define the structural environment of E314. The residue is nested in a hydrophobic environment that probably elevates its pKa, accounting for the pH dependency of drug efflux that we report in this work. Functional analyses of wild-type and mutant proteins in cells and proteoliposomes support our proposal for the mechanistic role of E314 in proton-coupled ethidium transport. As the carboxylate is known to participate in proton translocation by secondary-active transporters, our observations suggest that this substituent can play a similar role in the activity of ABC transporters. PMID- 16040837 TI - Recent developments in the evaluation of glomerular filtration rate: is there a place for beta-trace? PMID- 16040838 TI - Discovering rare variants by use of melting temperature shifts seen in melting curve analysis. PMID- 16040839 TI - Diagnostic proteomics: back to basics? PMID- 16040840 TI - Point: the metabolic syndrome still lives. PMID- 16040841 TI - Counterpoint: just being alive is not good enough. PMID- 16040842 TI - Comparison of the sensitivity of 2 automated immunoassays with immunofixation electrophoresis for detecting urine Bence Jones proteins. PMID- 16040843 TI - Straightforward procedure for internal control of real-time reverse transcription amplification assays. PMID- 16040844 TI - Identification of 19 new metabolites induced by abnormal amino acid conjugation in isovaleric acidemia. PMID- 16040845 TI - Relationship of plasma homocysteine with the severity of chronic heart failure. PMID- 16040846 TI - Evaluation of oxidative stress in serum of critically ill patients by a commercial assay and gas chromatography-mass spectrometry. PMID- 16040847 TI - Real-time reverse transcription-PCR quantification of vascular endothelial growth factor splice variants. PMID- 16040848 TI - Comparing whole-genome amplification methods and sources of biological samples for single-nucleotide polymorphism genotyping. PMID- 16040849 TI - LightTyper assay with locked-nucleic-acid-modified oligomers for genotyping of the toll-like receptor 4 polymorphisms A896G and C1196T. PMID- 16040850 TI - Comparison of HPLC and capillary electrophoresis for confirmatory testing of the alcohol misuse marker carbohydrate-deficient transferrin. PMID- 16040851 TI - Factor V null mutation affecting the Roche LightCycler factor V Leiden assay. PMID- 16040852 TI - Gas chromatographic-mass spectrometric analysis for measurement of p-cresol and its conjugated metabolites in uremic and normal serum. PMID- 16040853 TI - Detection of circulating prostate-specific antigen-secreting cells in prostate cancer patients. PMID- 16040854 TI - Plasma 8-isoprostane concentrations in patients with age-related cataracts. PMID- 16040855 TI - Cell-free DNA in serum and plasma: comparison of ELISA and quantitative PCR. PMID- 16040856 TI - Genotyping of the angiotensin I-converting enzyme gene insertion/deletion polymorphism by the TaqMan method. PMID- 16040857 TI - Pyrosequencing analysis of thrombosis-associated risk markers. PMID- 16040858 TI - Novel RET mutation produces a truncated RET receptor lacking the intracellular signaling domain in a 3-generation family with Hirschsprung disease. PMID- 16040859 TI - Homocysteine stability in heparinized plasma stored in a gel separator tube. PMID- 16040860 TI - Sensitive allele-specific PCR assay able to detect FGFR3 mutations in tumors and urine from patients with urothelial cell carcinoma of the bladder. PMID- 16040861 TI - Relationship between serum copper, ceruloplasmin, and non-ceruloplasmin-bound copper in routine clinical practice. PMID- 16040862 TI - Atypical melting curve resulting from genetic variation in the 3' untranslated region at position 20218 in the prothrombin gene analyzed with the LightCycler factor II (prothrombin) G20210A assay. PMID- 16040863 TI - Improved method for isolating cell-free DNA. PMID- 16040864 TI - Blood sampling: is fasting properly defined? PMID- 16040865 TI - Effects of prolonged ambient storage of sodium fluoride/heparin specimens on plasma homocysteine. PMID- 16040866 TI - Measurement of 25-hydroxyvitamin D by the Nichols ADVANTAGE, DiaSorin LIAISON, DiaSorin RIA, and liquid chromatography-tandem mass spectrometry. PMID- 16040867 TI - A perspective on perspectives. PMID- 16040868 TI - Circumcision for preventing urinary tract infections in boys: North American view. PMID- 16040869 TI - Circumcision for preventing urinary tract infection in boys: European view. PMID- 16040870 TI - Skull x rays, CT scans, and making a decision in head injury. PMID- 16040871 TI - Parenthood should not be regarded as a right. AB - In September 2004 the BBC broadcast a debate in the Straw Poll series about whether parenthood should be regarded as a right. The motion was carried by a large majority both among the audience present at the recording, and among listeners who were invited to telephone in with their votes, and views at a subsequent feedback programme. The arguments are presented here, in order to stimulate debate among readers. PMID- 16040872 TI - "Sour skin" in Darfur, Sudan. PMID- 16040873 TI - Parenthood should be regarded as a right. PMID- 16040874 TI - A cluster randomised intervention trial of asthma clubs to improve quality of life in primary school children: the School Care and Asthma Management Project (SCAMP). AB - AIM: To evaluate the effectiveness of a programme of asthma clubs in improving quality of life in primary school children with asthma. METHODS: A cluster randomised intervention trial was undertaken in 22 primary schools within the urban area of south and east Belfast, Northern Ireland. Schools were randomised in pairs to immediate or delayed groups. The study subjects comprised 173 children aged 7-11 years whose parents had notified the school of their asthma diagnosis. Children attended school based weekly clubs over an 8 week period. The main outcome measures were the interview administered Paediatric Quality of Life Questionnaire scores, ranging from 1 (worst) to 7 (best), spirometry, and inhaler technique. RESULTS: Over 15 weeks, small but non-significant improvements in the overall quality of life score (mean 0.20; 95% confidence interval (CI) -0.20 to 0.61) and in each of its three components, activity limitation (0.20; -0.43 to 0.84), symptoms (0.23; -0.23 to 0.70), and emotional function (0.17; -0.18 to 0.52), were observed in the immediate compared with the delayed group. Inhaler technique at week 16 was markedly better in the immediate group, with 56% having correct technique compared with 15% in the delayed group. No significant effect of the intervention on spirometry results could be demonstrated. CONCLUSION: This primary school based asthma education programme resulted in sustained improvements in inhaler technique, but changes in quality of life scores were not significant. PMID- 16040875 TI - Does carbon dioxide retention during exercise predict a more rapid decline in FEV1 in cystic fibrosis? AB - BACKGROUND: Carbon dioxide (CO2) retention during exercise is uncommon in mild to moderate lung disease in cystic fibrosis (CF). The ability to deal with increased CO2 is dependent on the degree of airflow limitation and inherent CO2 sensitivity. CO2 retention (CO2R) can be defined as a rise in P(ET)CO2 tension of > or =5 mm Hg with exercise together with a failure to reduce P(ET)CO2 tension after peak work by at least 3 mm Hg by the termination of exercise. AIM: To ascertain if carbon dioxide retention during exercise is associated with more rapid decline in lung function. METHODS: Annual spirometric and exercise data from 58 children aged 11-15 years, with moderate CF lung disease between 1996 and 2002 were analysed. RESULTS: The mean FEV1 at baseline for the two groups was similar; the CO2R group (n = 15) was 62% and the non-CO2 retention group (CO2NR) was 64% (n = 43). The decline in FEV1 after 12 months was -3.2% (SD 1.1) in the CO2R group and -2.3% (SD 0.9) in the CO2NR group. The decline after 24 months was -6.3% (SD 1.3) and -1.8% (SD 1.1) respectively. After 36 months, the decline in FEV1 was -5.3% (SD 1.2) and -2.6% (SD 1.1) respectively. The overall decline in lung function was 14.8% (SD 2.1) in the CO2R group and 6.7% (SD 1.8) in the CO2NR group. Using the primary outcome measure as a decline in FEV(1) of >9%, final multivariate analysis showed that the relative risks for this model were (95% CIs in parentheses): DeltaP(ET)CO2 11.61 (3.41 to 24.12), peak VO2 1.23 (1.10 to 1.43), and initial FEV(1) 1.14 (1.02 to 1.28). CONCLUSION: Results show that the inability to defend carbon dioxide during exercise is associated with a more rapid decline in lung function. PMID- 16040876 TI - Growth on stimulant medication; clarifying the confusion: a review. AB - AIMS: To get an overview of the studies of growth in height in children with attention deficit hyperactivity disorder (ADHD) treated with stimulant medication, to establish the consistencies and to try to resolve the discrepancies. METHODS: Twenty nine studies were reviewed following a Medline search: 22 related to children, six to late adolescents or adults, and one to children and adults. RESULTS: Children: Eleven studies gave results consistent with height attenuation on stimulant medication: eight were longitudinal, one was cross-sectional, and two showed growth rebound on ceasing medication. Studies with negative findings were inadequately powered (n = 3), lacked controls or statistical analysis (n = 3), measured height velocity without reference to treatment duration (n = 2), or used inappropriate growth parameters (n = 1), controls (n = 1), or normative data (n = 1). Late adolescents/adults treated with stimulant medication in childhood: Two studies associated childhood gastrointestinal side effects with attenuated late adolescent or adult height; all six cross-sectional studies had negative findings. The methodologies varied widely but there was some consistency in the degree of attenuation shown in studies with positive findings. The most sensitive methods analysed the changes in z-scores (standard deviation scores) or calculated the height deficits from paired measurements taken before and after an initial period of treatment with stimulant medication. The height deficit amounted to approximately 1 cm/year during the first 1-3 years of treatment. CONCLUSIONS: Further research is needed into the causal mechanisms, the rate of physical maturation, and the long term implications for final stature. PMID- 16040877 TI - How international are leading general paediatric journals? AB - The composition of the editorial boards of leading general paediatric journals was assessed. The board members' country of current affiliation was classified according to the Human Development Index. The results showed that only a very small number of the board members are based in the developing world. PMID- 16040878 TI - Secular trends in waist circumference in Spanish adolescents, 1995 to 2000-02. AB - Two studies, in 1995 and 2000-02, were compared to assess changes in waist circumference in adolescents. Between the two time periods, waist circumference increased significantly in males at 13 years and in females at 14 years. Significant changes in waist circumference were observed during the study period; the rates of change were 0.53 and 0.86 cm/y in boys and 0.67 and 0.87 cm/y in girls. Future morbidity in adolescents may be affected due to accumulation of excess central fat. PMID- 16040879 TI - The "Lasso-o" tape: stretchability and observer variability in head circumference measurement. AB - Head circumference is an important clinical measurement in children. The stretchability of the Lasso-o tape and the intra- and inter-observer reproducibility of measurements using it were investigated. Old Lasso-o tapes stretch significantly. The intra-class coefficients were 0.999 for intra-observer and 0.979 for inter-observer measurements. Nonetheless in 9% of measurements inter-observer variability was over 1 cm. PMID- 16040880 TI - The implications of the David Glass case for future clinical practice in the UK. AB - A recent decision of the European Court of Human Rights (ECtHR) raises issues of considerable importance to medical practitioners and paediatricians in particular. The case concerns the parental right to withhold consent to medical intervention that doctors believe to be necessary in a child's best interests. The dramatic facts of this case (in which a boy's family felt they had to fight for his life) has significant repercussions for clinical practice. This is discussed in the light of previous and recent cases that have involved babies, infants and children. The worrying trend to use the Courts to resolve these difficult clinical cases is discussed. PMID- 16040881 TI - Unusual foreign body detected on routine dental radiograph. PMID- 16040882 TI - Improving the management of atopic disease. AB - Asthma, wheeze, eczema, and, to a certain extent, rhinitis are very common conditions among children. The prevalence of allergic disease in the general population has increased alarmingly over the past 25 years, particularly in Western industrialised countries. However, it is important to remember that the symptoms often associated with allergy can have other aetiologies. Evidence suggests that in most circumstances, only 30-40% of chronic allergic-type symptoms are due to allergy. Accurate diagnosis of the presence of allergy is therefore an important issue, particularly given the interventions that such a diagnosis may initiate. In this review, we examine management options for allergy, provide the evidence as to what proportions of patients with common allergic-type symptoms are actually allergic, and list other causes of such symptoms. The importance of allergy testing and the options available are described, particularly with reference to the role of the non-allergist. PMID- 16040883 TI - How has research in the last five years changed my clinical practice? AB - The first instruction to examination candidates is to read and answer the question actually set. Doing so in this case leads to the following CONCLUSIONS: how research has changed my clinical practice includes the act of doing research, as well as reading about the work of others. Thus, this article refers to my own clinical practice (tertiary referral paediatric respiratory medicine in a setting where we do not service an accident and emergency department), rather than that of others. This means excluding important conditions such as acute croup and uncomplicated community acquired pneumonia. I should write about what has changed my practice, not what other people think I ought to have changed. So this will be a personal view, limited to research published in a peer review format at the time of writing. I shall also assume that change is an ongoing process, so I shall include change in progress, provided it is supported by published literature. PMID- 16040884 TI - Principles of evidence based medicine. AB - Health care professionals are increasingly required to base clinical decisions on the best available evidence. Evidence based medicine (EBM) is a systematic approach to clinical problem solving which allows the integration of the best available research evidence with clinical expertise and patient values. This paper explains the concept of EBM and introduces the five step EBM model: formulation of answerable clinical questions; searching for evidence; critical appraisal; applicability of evidence; evaluation of performance. Subsequent articles will focus on the principles and critical appraisal of randomised controlled trials, systematic reviews, and meta-analyses, and provide a practical demonstration of the five step EBM model using a real life clinical scenario. PMID- 16040885 TI - Understanding randomised controlled trials. AB - The hierarchy of evidence in assessing the effectiveness of interventions or treatments is explained, and the gold standard for evaluating the effectiveness of interventions, the randomised controlled trial, is discussed. Issues that need to be considered during the critical appraisal of randomised controlled trials, such as assessing the validity of trial methodology and the magnitude and precision of the treatment effect, and deciding on the applicability of research results, are discussed. Important terminologies such as randomisation, allocation concealment, blinding, intention to treat, p values, and confidence intervals are explained. PMID- 16040886 TI - Understanding systematic reviews and meta-analysis. AB - This review covers the basic principles of systematic reviews and meta-analyses. The problems associated with traditional narrative reviews are discussed, as is the role of systematic reviews in limiting bias associated with the assembly, critical appraisal, and synthesis of studies addressing specific clinical questions. Important issues that need to be considered when appraising a systematic review or meta-analysis are outlined, and some of the terms used in the reporting of systematic reviews and meta-analyses--such as odds ratio, relative risk, confidence interval, and the forest plot--are introduced. PMID- 16040887 TI - Evidence in practice. AB - A clinical scenario is used to illustrate how the principles outlined in the previous articles in the series could be applied to help improve patient care. A practical demonstration of the art of formulating answerable clinical questions, finding evidence, critically appraising evidence, and putting evidence into practice is provided. The importance of integrating evidence with patient's preferences, and taking account of issues such as availability of interventions, costs, and so on is discussed. Finally, some of the issues involved in the development of evidence based policies within clinical teams are outlined. PMID- 16040888 TI - Lethal cardiomyopathy in epidermolysis bullosa associated with amitriptyline. AB - There are previous reports of dilated cardiomyopathy (DCM) in recessive dystrophic epidermolysis bullosa (RDEB), a debilitating blistering skin disorder. The pathogenesis of DCM in RDEB remains uncertain, although dietary deficiency of selenium and carnitine have been implicated. A 6 year old girl with RDEB who died of DCM is reported; attention is drawn to the possible role of two potentially cardiotoxic drugs, amitriptyline and cisapride. PMID- 16040889 TI - Palivizumab prophylaxis in haemodynamically significant congenital heart disease. PMID- 16040890 TI - Developmental delay versus developmental impairment. PMID- 16040891 TI - Rejoinder to Eigenmann PA, Haengelli CA, Food colourings and preservatives- allergy and hyperactivity (Lancet 2004; 364:823-4) and an erratum. PMID- 16040892 TI - The Royal College of Paediatrics and Child Health flagship meeting: is it value for money? PMID- 16040893 TI - Meningitis is a common cause of convulsive status epilepticus. PMID- 16040894 TI - Radiofrequency-enhanced vascular gene transduction and expression for intravascular MR imaging-guided therapy: feasibility study in pigs. AB - PURPOSE: To evaluate the feasibility of radiofrequency (RF)-enhanced vascular gene transduction and expression by using a magnetic resonance (MR) imaging heating guidewire as an intravascular heating vehicle during MR imaging-guided therapy. MATERIALS AND METHODS: The institutional committee for animal care and use approved the experimental protocol. The study included in vitro evaluation of the use of RF energy to enhance gene transduction and expression in vascular cells, as well as in vivo validation of the feasibility of intravascular MR imaging-guided RF-enhanced vascular gene transduction and expression in pig arteries. For in vitro experiments, approximately 10(4) vascular smooth muscle cells were seeded in each of four chambers of a cell culture plate. Next, 1 mL of a green fluorescent protein gene (gfp)-bearing lentivirus was added to each chamber. Chamber 4 was heated at approximately 41 degrees C for 15 minutes by using an MR imaging-heating guidewire connected to a custom RF generator. At day 6 after transduction, the four chambers were examined and compared at confocal microscopy to determine the efficiency of gfp transduction and expression. For the in vivo experiments, a lentivirus vector bearing a therapeutic gene, vascular endothelial growth factor 165 (VEGF-165), was transferred by using a gene delivery balloon catheter in 18 femoral-iliac arteries (nine artery pairs) in domestic pigs and Yucatan pigs with atherosclerosis. During gene infusion, one femoral-iliac artery in each pig was heated to approximately 41 degrees C with RF energy transferred via the intravascular MR imaging-heating guidewire, while the contralateral artery was not heated (control condition). At day 6, the 18 arteries were harvested for quantitative Western blot analysis to compare VEGF 165 transduction and expression efficiency between RF-heated and nonheated arterial groups. RESULTS: Confocal microscopy showed gfp expression in chamber 4 that was 293% the level of expression in chamber 1 (49.6% +/- 25.8 vs 16.8% +/- 8.0). Results of Western blot analysis showed VEGF-165 expression for normal arteries in the RF-heated group that was 300% the level of expression in the nonheated group (70.4 arbitrary units [au] +/- 107.1 vs 23.5 au +/- 29.8), and, for atherosclerotic arteries in the RF-heated group, 986% the level in the nonheated group (129.2 au +/- 100.3 vs 13.1 au +/- 4.9). CONCLUSION: Simultaneous monitoring and enhancement of vascular gene delivery and expression is feasible with the MR imaging-heating guidewire. PMID- 16040895 TI - What parameters are most accurate in predicting appropriate technique factors for CT scanning? PMID- 16040896 TI - Reinventing radiology in the digital age: part I. The all-digital department. PMID- 16040898 TI - Biomedical Imaging Research Opportunities Workshop II: report and recommendations. PMID- 16040899 TI - Ethics and the limits of neonatal viability. PMID- 16040900 TI - How I do it: evaluating renal masses. AB - With modern computed tomography (CT) and magnetic resonance (MR) imaging equipment, the diagnosis of most renal masses is usually straightforward and accurate. The major question to be answered is whether the mass represents a surgical or nonsurgical lesion or, in some cases, if follow-up studies are necessary. This evaluation usually can be accomplished if a high-quality examination is performed, if the clinical history of the patient is kept in mind, if conditions that mimic a renal neoplasm are considered and excluded, and if there is an awareness of the potential pitfalls and limitations of CT and MR imaging. In this article, the authors present their technique in the performance of CT and MR imaging examinations, summarize their approach to the diagnosis of renal masses, review the imaging findings in these lesions, and stress the limitations in renal mass diagnosis. PMID- 16040901 TI - Computer-aided detection with screening mammography in a university hospital setting. AB - PURPOSE: To prospectively assess the effect of computer-aided detection (CAD) on screening mammogram interpretation in an academic medical center to determine if the outcome is different than that previously reported for community practices. MATERIALS AND METHODS: Institutional review board approval was granted, and informed consent was waived. During a 19-month period, 8682 women (median age, 54 years; range, 33-95 years) underwent screening mammography. Each mammogram was interpreted by one of seven radiologists, followed by immediate re-evaluation of the mammogram with CAD information. Each recalled case was classified as follows: radiologist perceived the finding and CAD marked it, radiologist perceived the finding and CAD did not mark it, or CAD prompted the radiologist to perceive the finding and recall the patient. Lesion type was also recorded. Recalled patients were tracked to determine the effect of CAD on recall and biopsy recommendation rates, positive predictive value (PPV) of biopsy, and cancer detection rate. A 95% confidence interval was calculated for cancer detection rate. Pathologic examination was performed for all cancers. RESULTS: Of 8682 patients, 863 (9.9%) with 960 findings were recalled for further work-up (Breast Imaging Reporting and Data System category 0). After further diagnostic imaging, it was recommended that biopsy or aspiration be performed for 181 of 960 findings (19%); 165 interventions were confirmed to have been performed. Twenty-nine cancers were found in this group, with a PPV for biopsy of 18% (29 of 165 findings) and a cancer detection rate of 3.3 per 1000 screening mammograms (29 of 8682 patients). CAD-prompted recalls contributed 8% (73 of 960 findings) of total recalled findings and 7% (two of 29 lesions) of cancers detected. Of 29 cancers (59%), 17 manifested as masses and 12 (41%) were microcalcifications. Ten (34%) cancers were ductal carcinoma in situ, and the remaining cancers had an invasive component. Both cancers found with CAD manifested as masses, and both were invasive ductal carcinoma. CONCLUSION: Prospective clinical use of CAD in a university hospital setting resulted in a 7.4% increase (from 27 to 29) in cancers detected. Both cancers were nonpalpable masses. PMID- 16040902 TI - Solid breast masses: classification with computer-aided analysis of continuous US images obtained with probe compression. AB - PURPOSE: To prospectively evaluate the accuracy of continuous ultrasonographic (US) images obtained during probe compression and computer-aided analysis for classification of biopsy-proved (reference standard) benign and malignant breast tumors. MATERIALS AND METHODS: This study was approved by the local ethics committee, and informed consent was obtained from all included patients. Serial US images of 100 solid breast masses (60 benign and 40 malignant tumors) were obtained with US probe compression in 86 patients (mean age, 45 years; range, 20 67 years). After segmentation of tumor contours with the level-set method, three features of strain on tissue from probe compression--contour difference, shift distance, area difference--and one feature of shape--solidity-were computed. A maximum margin classifier was used to classify the tumors by using these four features. The Student t test and receiver operating characteristic curve analysis were used for statistical analysis. RESULTS: The mean values of contour difference, shift distance, area difference, and solidity were 3.52% +/- 2.12 (standard deviation), 2.62 +/- 1.31, 1.08% +/- 0.85, and 1.70 +/- 1.85 in malignant tumors and 9.72% +/- 4.54, 5.04 +/- 2.79, 3.17% +/- 2.86, and 0.53 +/- 0.63 in benign tumors, respectively. Differences with P < .001 were statistically significant for all four features. Area under the receiver operating characteristic curve (A(Z)) values for contour difference, shift distance, area difference, and solidity were 0.88, 0.85, 0.86, and 0.79, respectively. The A(Z) value of three features of strain was significantly higher than that of the feature of shape (P < .01). The accuracy, sensitivity, specificity, and positive and negative predictive values of US classifications that were based on values for these four features were 87.0% (87 of 100), 85% (34 of 40), 88% (53 of 60), 83% (34 of 41), and 90% (53 of 59), respectively, with an A(Z) value of 0.91. CONCLUSION: Continuous US images obtained with probe compression and computer aided analysis can aid in classification of benign and malignant breast tumors. PMID- 16040903 TI - Adding in vivo quantitative 1H MR spectroscopy to improve diagnostic accuracy of breast MR imaging: preliminary results of observer performance study at 4.0 T. AB - PURPOSE: To determine whether the addition of in vivo quantitative hydrogen 1 (1H) magnetic resonance (MR) spectroscopy can improve the radiologist's diagnostic accuracy in interpreting breast MR images to distinguish benign from malignant lesions. MATERIALS AND METHODS: The study was approved by the institutional review board and, where appropriate, was compliant with the Health Insurance Portability and Accountability Act. All patients provided written informed consent. Fifty-five breast MR imaging cases-one lesion each in 55 patients aged 24-66 years with biopsy-confirmed findings-were retrospectively evaluated by four radiologists. Patients were examined with contrast material enhanced fat-suppressed T1-weighted 4.0-T MR imaging. The concentration of total choline-containing compounds (tCho) was quantified by using single-voxel 1H MR spectroscopy. For each case, the radiologists were asked to give the percentage probability of malignancy, the Breast Imaging and Reporting Data System category, and a recommendation for patient treatment. Two interpretations were performed for each case: The initial interpretation was based on the lesion's morphologic features and time-signal intensity curve, and the second interpretation was based on the lesion's morphologic features, time-signal intensity curve, and tCho concentration. Receiver operating characteristic (ROC), Wilcoxon signed rank, kappa statistic, and accuracy (based on the area under the ROC curve) analyses were performed. RESULTS: Of the 55 lesions evaluated, 35 were invasive carcinomas and 20 were benign. The addition of 1H MR spectroscopy resulted in higher sensitivity, specificity, accuracy, and interobserver agreement for all four radiologists. More specifically, two of the four radiologists achieved a significant improvement in sensitivity (P=.03, P=.03), and all four radiologists achieved a significant improvement in accuracy (P = .01, P = .05, P = .009, P < .001). CONCLUSION: Current study results suggest that the addition of quantitative 1H MR spectroscopy to the breast MR imaging examination may help to improve the radiologist's ability to distinguish benign from malignant breast lesions. PMID- 16040905 TI - Case 85: pelvic actinomycosis in association with an intrauterine device. PMID- 16040906 TI - Radiofrequency ablation: importance of background tissue electrical conductivity- an agar phantom and computer modeling study. AB - PURPOSE: To determine whether radiofrequency (RF)-induced heating can be correlated with background electrical conductivity in a controlled experimental phantom environment mimicking different background tissue electrical conductivities and to determine the potential electrical and physical basis for such a correlation by using computer modeling. MATERIALS AND METHODS: The effect of background tissue electrical conductivity on RF-induced heating was studied in a controlled system of 80 two-compartment agar phantoms (with inner wells of 0.3%, 1.0%, or 36.0% NaCl) with background conductivity that varied from 0.6% to 5.0% NaCl. Mathematical modeling of the relationship between electrical conductivity and temperatures 2 cm from the electrode (T2cm) was performed. Next, computer simulation of RF heating by using two-dimensional finite-element analysis (ETherm) was performed with parameters selected to approximate the agar phantoms. Resultant heating, in terms of both the T2cm and the distance of defined thermal isotherms from the electrode surface, was calculated and compared with the phantom data. Additionally, electrical and thermal profiles were determined by using the computer modeling data and correlated by using linear regression analysis. RESULTS: For each inner compartment NaCl concentration, a negative exponential relationship was established between increased background NaCl concentration and the T2cm (R2= 0.64-0.78). Similar negative exponential relationships (r2 > 0.97%) were observed for the computer modeling. Correlation values (R2) between the computer and experimental data were 0.9, 0.9, and 0.55 for the 0.3%, 1.0%, and 36.0% inner NaCl concentrations, respectively. Plotting of the electrical field generated around the RF electrode identified the potential for a dramatic local change in electrical field distribution (ie, a second electrical peak ["E-peak"]) occurring at the interface between the two compartments of varied electrical background conductivity. Linear correlations between the E-peak and heating at T2cm (R2= 0.98-1.00) and the 50 degrees C isotherm (R2= 0.99-1.00) were established. CONCLUSION: These results demonstrate the strong relationship between background tissue conductivity and RF heating and further explain electrical phenomena that occur in a two-compartment system. PMID- 16040907 TI - T2-prepared steady-state free precession blood oxygen level-dependent MR imaging of myocardial perfusion in a dog stenosis model. AB - PURPOSE: To assess the ability of a T2-prepared steady-state free precession blood oxygen level-dependent (BOLD) magnetic resonance (MR) imaging sequence to depict changes in myocardial perfusion during stress testing in a dog stenosis model. MATERIALS AND METHODS: Study was approved by the institutional Animal Care and Use Committee. A hydraulic occluder was placed in the left circumflex coronary artery (LCX) in 10 dogs. Adenosine was administered intravenously to increase coronary blood flow, and stenosis was achieved in the LCX with the occluder. A T2-prepared two-dimensional steady-state free precession sequence was used for BOLD imaging at a spatial resolution of 1.5 x 1.2 x 5.0 mm3, and first pass perfusion images were acquired for visual comparison. Microspheres were injected to provide regional perfusion information. Mixed-effect regression analysis was performed to assess normalized MR signal intensity ratios and microsphere-measured perfusion differences. For the same data, 95% prediction intervals were calculated to determine the smallest perfusion change detectable. Means +/- standard deviations were calculated for myocardial regional comparison data. A two-tailed Student t test was used to determine if significant differences (P < .01) existed between different myocardial regions. RESULTS: Under maximal adenosine stress, MR clearly depicted stenotic regions and showed regional signal differences between the left anterior descending coronary artery (LAD)-fed myocardium and the stenosed LCX-fed myocardium. Visual comparisons with first-pass images were also excellent. Regional MR signal intensity differences between LAD and LCX-fed myocardium (1.24 +/- 0.08) were significantly different (P < .01) from differences between LAD and septal-fed myocardium (1.02 +/- 0.07), which was in agreement with microsphere-measured flow differences (LAD/LCX, 3.38 +/- 0.83; LAD/septal, 1.26 +/- 0.49). The linear mixed-effect regression model showed good correlation (R = 0.79) between MR differences and microsphere measured flow differences. CONCLUSION: On T2-prepared steady-state free precession BOLD MR images in dogs, signal intensity differences were linearly related to flow differences in myocardium, with a high degree of correlation. SUPPLEMENTAL MATERIAL: radiology.rsnajnls.org/cgi/content/full/236/2/503/DC1 PMID- 16040908 TI - Gadomer-enhanced MR imaging in the detection of microvascular obstruction: alleviation with nicorandil therapy. AB - PURPOSE: To evaluate Gadomer-enhanced magnetic resonance (MR) imaging in the quantification of small microvascular obstruction regions and determine if nicroandil alleviates the formation of microvascular obstruction. MATERIALS AND METHODS: Approval of the institutional committee on animal research was obtained, and this study complied with guidelines for care and use of animals. Rats underwent coronary artery occlusion and reperfusion. After 24 hours, Gadomer enhanced T1-weighted spin-echo MR imaging was used to define microvascular obstruction in animals in control and nicorandil groups. Sequential MR images obtained at two midventricular levels were acquired to measure microvascular obstruction and ischemically injured regions and monitor diffusive and/or convective transport of Gadomer in microvascular obstruction regions. Two investigators working in consensus and using threshold signal intensity measured differentially enhanced regions. Left-ventricular (LV) end-systolic and end diastolic MR images obtained at the same two midventricular levels were used to measure regional wall thickening and systolic reduction in LV relative volumes. Agreement and correlation between MR imaging and postmortem data were determined with Bland-Altman and linear regression analyses. Animals were sacrificed 3 minutes after intravenous injection of blue dye. RESULTS: On Gadomer-enhanced MR images, two differentially enhanced regions were observed in ischemically injured myocardium, namely, the hypoenhanced region and the surrounding hyperenhanced region. Hypoenhanced regions at MR imaging and unstained regions at blue dye administration were identical 3 minutes after administration (17% +/- 1 and 17% +/- 2; P = .6; r = 0.98). In the control group, Gadomer provided a prolonged imaging window (eg, 6 minutes) for accurately quantifying small microvascular obstruction regions. Microvascular obstruction was observed in all animals in the control group and 27% of animals in the nicorandil group. Microvascular obstruction regions were smaller in the nicorandil group (eg, 3% +/- 1) than in the control group (eg, 17% +/- 2) (P < .001). Hyper- and hypoenhanced regions were also smaller (eg, 20% +/- 2) in rats in the nicorandil group than in those in the control group (37% +/- 4, P < .001). Improvement in LV function in the nicorandil group is likely related to alleviation and reduction in infarct size. CONCLUSION: Gadomer-enhanced MR imaging can be used to quantify small microvascular obstruction regions 24 hours after reperfusion. Intravenous therapy with nicorandil reduces formation of microvascular obstruction regions. PMID- 16040909 TI - Extracolonic abnormalities discovered incidentally at CT colonography in a male population. AB - PURPOSE: To evaluate prospectively the prevalence of incidental extracolonic findings at computed tomographic (CT) colonography and to estimate the cost of their imaging work-up in male patients with high and those with average risk of colorectal cancer. MATERIALS AND METHODS: This study was approved by the institutional review board, and informed consent was obtained from all patients. The study was compliant with requirements of the Health Insurance Portability and Accountability Act. CT colonography was performed in 500 men (mean age, 62.5 years). Of these patients, 194 (38.8%) were at average risk for colorectal cancer and presented for routine screening. The other 306 (61.2%) were at high risk for colorectal cancer. Extracolonic findings were recorded and categorized as either clinically important or clinically unimportant. Clinically important findings were defined as those that necessitated further diagnostic studies or medical or surgical follow-up. The cost of additional imaging required to further characterize important lesions was estimated. Chart review was performed (mean length of follow-up, 3.6 years) to determine whether any important findings were missed at CT colonography. The Fisher exact test was used to determine whether there was a difference between the percentages of average- and high-risk patients with extracolonic findings. RESULTS: Of the 500 patients in the study, 315 (63.0%) had extracolonic findings, and 45 (9.0%) had clinically important extracolonic findings. Of the 596 extracolonic findings identified, 50 (8.4%) were thought to be clinically important. The mean additional cost to work up important findings was $28.12 per CT colonographic examination. There were no significant differences between average-risk and high-risk patients in the percentages of extracolonic findings (P = .25) or clinically important extracolonic findings (P = .11). CONCLUSION: A substantial number of both average and high-risk patients undergoing CT colonography will be found to have clinically important extracolonic findings. There was no increased morbidity or mortality associated with the additional evaluation of extracolonic findings. The cost of evaluating these lesions is low, given the potential for positive effects on patient care. PMID- 16040910 TI - Comparison of US and unenhanced multi-detector row CT in patients suspected of having acute appendicitis. AB - PURPOSE: To prospectively compare the diagnostic performance of ultrasonography (US) and unenhanced multi-detector row computed tomography (CT) in patients suspected of having acute appendicitis by using surgery or clinical follow-up as the reference standard. MATERIALS AND METHODS: The institutional review board approved the research protocol. Written informed consent was obtained from all patients or, for those who were adolescents, from their parents. Ninety-four patients (59 female and 35 male patients) aged 16-81 years (mean, 38 years) who were suspected of having acute appendicitis underwent both US and unenhanced multi-detector row CT of the entire abdomen. The examinations were performed within 1-2 hours of each other. US and CT images were obtained and prospectively interpreted by a different radiologist from a group of abdominal radiologists or a group of residents and general radiologists. Radiologists proposed an overall diagnosis and an alternative diagnosis. Data from US and CT were compared, and the definite diagnosis was established with surgical findings (n = 40) or results of clinical follow-up (n = 54) as the reference standard. Comparisons were made for each group of radiologists and the patient's age, body mass index (BMI), and sex. Proportion comparisons were made by using the Pearson chi2 test or the Fisher exact test. Continuous variables were compared between groups with the Mann-Whitney U test. RESULTS: Thirty patients had definite appendicitis. The sensitivity, specificity, positive and negative predictive values, and accuracy were not significantly different between US and CT or between groups of radiologists (P values ranged from .389 to >.99), regardless of the patient's BMI (P values ranged from .073 to >.99). Misclassifications were compared with the definite alternative diagnosis and were not significantly different between US and CT or between groups of radiologists (P = .061-.592), regardless of patient age (P = .875) or sex (P = .151 and >.99 for male and female patients, respectively). The frequency of inconclusive examinations, however, was significantly higher with US than with CT, regardless of radiologist experience (P = .020 and <.001, respectively). CONCLUSION: Although the diagnostic performances of US and multi-detector row CT are comparable, more inconclusive images were obtained with US. PMID- 16040911 TI - Superficial esophageal cancer: esophagographic findings correlated with histopathologic findings. AB - PURPOSE: To retrospectively determine and evaluate the findings of superficial esophageal cancer at esophagography and to correlate the esophagographic findings with the depth of tumor invasion. MATERIALS AND METHODS: The institutional review boards required neither their approval nor informed patient consent for this retrospective study. One hundred thirteen patients with superficial esophageal cancer who underwent esophagectomy at three institutions were included in this study. Double-contrast esophagograms were reviewed independently by two reviewers. For assessment of histopathologic findings, pathology reports were reviewed. Findings at esophagography, including morphologic type of the lesion, lesion extent, presence or absence of elevated or depressed component, margin and extent of elevated or depressed component, presence or absence of nodularity, extent of nodularity, esophageal luminal narrowing, and esophageal wall rigidity, were compared between mucosal and submucosal cancers by using chi2, Fisher exact, and independent-sample t tests. RESULTS: Of 122 histopathologically proved superficial esophageal cancers in 113 patients, 100 (82%) were detected at esophagography. The most common morphologic type was the plaquelike form; 50 (50%) such lesions were depicted at esophagography. Morphologic types were significantly different between the mucosal and submucosal cancers (P < .001). Protruded and plaquelike lesions were more frequent among submucosal cancers, whereas most flat lesions were mucosal cancers. An elevated component (P < .001), a rigid esophageal wall (P < .001), and a lobulated or irregular margin of the elevated component (P = .023) were significantly more frequent among submucosal cancers. Also, total extent of the lesion (P < .001), size of the largest nodule (P < .001), and extent of nodularity (P = .036) were significantly larger in the submucosal cancers. CONCLUSION: In the evaluation of patients with superficial esophageal cancer, esophagography appears to be helpful for diagnosing the tumor and differentiating mucosal from submucosal cancers. PMID- 16040912 TI - Renal colic: comparison of use and outcomes of unenhanced helical CT for emergency investigation in 1998 and 2002. AB - PURPOSE: To determine retrospectively whether there had been any change between 1998 and 2002 in the use and outcome of computed tomography (CT) performed in the emergency department for patients presenting with symptoms of renal colic. MATERIALS AND METHODS: Approval from the Research Ethics Board was obtained, and informed consent was waived. All CT examinations ordered from the emergency department of a tertiary care hospital and performed from July to December 1998 and July to December 2002 were identified. Reports were reviewed, and results were categorized as either (a) positive for urinary tract calculus disease (category I), (b) indicative of an alternate diagnosis (category II), or (c) negative for findings to account for the patient's symptoms (category III). The corresponding emergency department charts were reviewed for urine dipstick results for hematuria and for patient history of stone disease. For statistical analysis, chi2 testing and odds ratios were used. RESULTS: During the 6-month period in 1998, 179 CT examinations were performed in patients who were admitted to the emergency department. During the same period in 2002, 234 CT examinations were performed. After correction for the total number of emergency department visits, it was determined that there was a relative increase of 21.3% (95% confidence interval: -0.0009, 0.47) in number of CT examinations performed in the emergency department. A total of 117 (65.4%) of 179 CT studies in 1998 and 153 (65.4%) of 234 CT studies in 2002 demonstrated renal calculus disease (category I), nine (5.0%) of 179 CT studies in 1998 and 17 (7.3%) of 234 CT studies in 2002 were used to identify an alternate diagnosis for patient symptoms (category II), and 53 (29.6%) of 179 CT studies in 1998 and 64 (27.4%) of 234 CT studies in 2002 were negative (category III). There were no significant differences between the rates of category I, II, or III results and the positivity rates for hematuria and urinary tract stone history during 1998 and 2002. CONCLUSION: Despite a definite trend of increased CT use during 1998 and 2002, there was no significant decrease in the rates of positive renal colic results or alternate diagnoses. PMID- 16040913 TI - Temporomandibular joint disk position assessed at coronal MR imaging in asymptomatic volunteers. AB - PURPOSE: To assess the normal position of the temporomandibular joint (TMJ) disk relative to the condyle by using coronal magnetic resonance (MR) imaging in asymptomatic volunteers. MATERIALS AND METHODS: This study was approved by the review committee for human research, and all subjects signed an informed consent form. Thirty symptom-free volunteers without histories of TMJ disorders underwent standardized clinical examinations. Afterward, bilateral sagittal oblique and coronal oblique MR images were acquired with the patient's mouth opened and closed. The coronal oblique opened- and closed-mouth images were analyzed by using computer software. The medial and lateral edges of both the TMJ disk and the condyle were marked for these imaging examinations by using the section through the posterior 3 mm of the disk. To eliminate the effect of different magnifications and/or distortions, the distance between these points was measured automatically and divided by the largest mediolateral dimensions of the condyle. To assess the reliability of the measurements, four observers evaluated the position of the disk in the coronal plane. To assess the changes in position of the posterior 3 mm of the disk in the coronal plane in the closed- and opened mouth positions, the Wilcoxon signed rank test for matched pairs was used. Interobserver measurement reliability was evaluated by using interclass correlation coefficients (ICCs). RESULTS: Analysis of the coronal closed-mouth disk position revealed a medial position of the TMJ disk relative to the condyle in 11 (21%) of 52 analyzed joints. In the opened-mouth position, the medial location of the disk was more frequent: 29 (85%) of 34 analyzed joints exhibited a medial position of the disk relative to the condyle in this plane. This increasingly medial position of the disk was statistically significant (P < or = .001). Measurement reliability assessment revealed sufficient results (ICC > or = 0.7). CONCLUSION: At both closed- and opened-mouth MR imaging, a medially located TMJ disk seems to be within the normal range of variation. The disk seems to shift even more medially when the mouth is opened. PMID- 16040914 TI - Comparison of different body size parameters for individual dose adaptation in body CT of adults. AB - PURPOSE: To investigate prospectively which of several body size parameters are suitable for individual dose adaptation in body computed tomography (CT) of adults. MATERIALS AND METHODS: Three body regions (thorax, abdomen, pelvis) were scanned exclusively for clinical reasons, with institutional ethical approval and informed consent. For each of the three regions, 50 men and 50 women (aged 18-87 years) were studied (300 scans total). Individual x-ray properties for each scan were summarized with a water-equivalent diameter (Dw). Different body size parameters, based on weight, height, and shape, were correlated with Dw by using regression analysis. This resulted in Dw estimation errors of different magnitudes, indicated with 95% prediction intervals. The errors from weight were compared with those from each of the other body parameters by using comparison of variance in paired samples (P < .05). In addition, a topogram-based estimate for Dw was studied, which simulated an automated body size measurement. RESULTS: For the thorax, abdomen, and pelvis, mean Dw was 28.0, 29.1, and 29.3 cm, and estimation of Dw from weight enabled 95% prediction intervals of +/-2.5, +/-2.4, and +/-2.6 cm, respectively. Combinations of height and weight were only slightly more or even less exact than were measurements from only weight. Diameter-related parameters such as body circumference were similar to or better than weight. However, the topogram-based estimate was significantly more exact. CONCLUSION: Body weight and circumference enable suitable estimates for individual dose adaptation in body CT of adults if automated dose adaptation is not available. PMID- 16040915 TI - Evaluation of gas-filled microparticles and sonoporation as gene delivery system: feasibility study in rodent tumor models. AB - PURPOSE: To evaluate the feasibility of gene delivery mediated with diagnostic ultrasound and plasmid DNA (pDNA) encapsulated in gas-filled microparticles (GFMP) in rodent tumor models. MATERIALS AND METHODS: This study was performed according to a protocol approved by the regional animal research committee. The model plasmid UT651 (pUT651) that contained the Escherichia coli LacZ gene for beta-galactosidase was used to demonstrate the feasibility of ultrasound-mediated gene delivery in CC531 liver tumors in rats. In preliminary experiments, a single injection of pUT651-containing GFMP was administered intraarterially (n=4) or intravenously (n=6) with simultaneous sonication (color Doppler mode, maximum mechanical index) of the GFMP passing through the capillaries of the tumors. All animals were sacrificed 2-5 days later, and liver tumors were examined for beta galactosidase expression histochemically. Subsequently, potential medical usefulness of this delivery system was tested in nude mice bearing Capan-1 tumors (adenocarcinoma of the human pancreas) by using the plasmid RC/CMV-p16 (pRC/CMV p16), which contains tumor suppressor gene p16. The tumor suppressor gene p16 is deleted in Capan-1 cells. Twenty-five tumor-bearing mice were classified into five groups (four to six mice per group, one treatment group, four control groups) at random. All mice were treated once weekly for 5 weeks with intravenous infusion of p16-containing GFMP or control substances with simultaneous tumor sonication with color Doppler mode ultrasound and maximum mechanical index or without ultrasound treatment. The therapeutic effect of p16 was measured as an increase in tumor volume doubling time. Data were analyzed with analysis of variance. Results were considered significant at the 5% critical level (P < .05). RESULTS: A clear expression of pDNA was found in tumors in rats treated with a combination of pUT651-containing GFMP and ultrasound; relevant controls showed a significantly lower expression of marker gene. The controlled ultrasound triggered release of pRC/CMV-p16 from GFMP leads to a strong tumor growth inhibition, which is significant (P < .002), compared with that in controls. CONCLUSION: A combination of GFMP and ultrasound provides an effective approach for nonviral gene therapy-based cancer treatment. PMID- 16040916 TI - Intramyocellular lipid quantification: repeatability with 1H MR spectroscopy. AB - PURPOSE: To prospectively determine the repeatability and variability of tibialis anterior intramyocellular lipid (IMCL) quantifications performed by using 1.5-T hydrogen 1 (1H) magnetic resonance (MR) spectroscopy in healthy subjects. MATERIALS AND METHODS: Institutional review board approval and written informed consent were obtained for this Health Insurance Portability and Accountability Act-compliant study. The authors examined the anterior tibial muscles of 27 healthy subjects aged 19-48 years (12 men, 15 women; mean age, 25 years) by using single-voxel short-echo-time point-resolved 1H MR spectroscopy. During a first visit, the subjects underwent 1H MR spectroscopy before and after being repositioned in the magnet bore, with voxels carefully placed on the basis of osseous landmarks. Measurements were repeated after a mean interval of 12 days. All spectra were fitted by using Java-based MR user interface (jMRUI) and LCModel software, and lipid peaks were scaled to the unsuppressed water peak (at 4.7 ppm) and the total creatine peak (at approximately 3.0 ppm). A one-way random-effects variance components model was used to determine intraday and intervisit coefficients of variation (CVs). A power analysis was performed to determine the detectable percentage change in lipid measurements for two subject sample sizes. RESULTS: Measurements of the IMCL methylene protons peak at a resonance of 1.3 ppm scaled to the unsuppressed water peak (IMCL(W)) that were obtained by using jMRUI software yielded the lowest CVs overall (intraday and intervisit CVs, 13.4% and 14.4%, respectively). The random-effects variance components model revealed that nonbiologic factors (equipment and repositioning) accounted for 50% of the total variability in IMCL quantifications. Power analysis for a sample size of 20 subjects revealed that changes in IMCL(W) of greater than 15% could be confidently detected between 1H MR spectroscopic measurements obtained on different days. CONCLUSION: 1H MR spectroscopy is feasible for repeatable quantification of IMCL concentrations in longitudinal studies of muscle metabolism. PMID- 16040917 TI - Determination of multiple sclerosis plaque size with diffusion-tensor MR Imaging: comparison study with healthy volunteers. AB - PURPOSE: To use diffusion-tensor magnetic resonance (MR) imaging to measure involvement of normal-appearing white matter (WM) immediately adjacent to multiple sclerosis (MS) plaques and thus redefine actual plaque size on diffusion tensor images through comparison with T2-weighted images of equivalent areas in healthy volunteers. MATERIALS AND METHODS: Informed consent was not required given the retrospective nature of the study on an anonymized database. The study complied with requirements of the Health Insurance Portability and Accountability Act. Twelve patients with MS (four men, eight women; mean age, 35 years) and 14 healthy volunteers (six men, eight women; mean age, 25 years) were studied. The authors obtained fractional anisotropy (FA) values in MS plaques and in the adjacent normal-appearing WM in patients with MS and in equivalent areas in healthy volunteers. They placed regions of interest (ROIs) around the periphery of plaques and defined the total ROIs (ie, plaques plus peripheral ROIs) as abnormal if their mean FA values were at least 2 standard deviations below those of equivalent ROIs within equivalent regions in healthy volunteers. The combined area of the plaque and the peripheral ROI was compared with the area of the plaque seen on T2-weighted MR images by means of a Student paired t test (P = .05). RESULTS: The mean plaque size on T2-weighted images was 72 mm2 +/- 21 (standard deviation). The mean plaque FA value was 0.285 +/- 0.088 (0.447 +/- 0.069 in healthy volunteers [P < .001]; mean percentage reduction in FA in MS plaques, 37%). The mean plaque size on FA maps was 91 mm2 +/- 35, a mean increase of 127% compared with the size of the original plaque on T2-weighted images (P = .03). CONCLUSION: A significant increase in plaque size was seen when normal appearing WM was interrogated with diffusion-tensor MR imaging. This imaging technique may represent a more sensitive method of assessing disease burden and may have a future role in determining disease burden and activity. PMID- 16040918 TI - Middle cerebral artery spasm after subarachnoid hemorrhage: detection with transcranial color-coded duplex US. AB - PURPOSE: To prospectively determine the accuracy of transcranial color-coded duplex ultrasonography (US) used alone and in conjunction with carotid artery US for diagnosis of middle cerebral artery (MCA) spasm, with intraarterial digital subtraction angiography (DSA) used as the reference standard. MATERIALS AND METHODS: The institutional ethics committee approved the study. Each patient, or members of the patient's family, gave informed consent. One hundred twenty consecutive patients (64 women, 56 men; mean age, 45.5 years +/- 13.6 [standard deviation]) were routinely referred for DSA after subarachnoid hemorrhage. Vasospasm was graded as mild (< or =25% reduction in vessel diameter), moderate (>25% to 50% reduction), or severe (>50% reduction). US was performed 2 hours or less before angiography. The ratio of flow velocity in the middle cerebral artery (V(MCA)) to flow velocity in the ipsilateral extracranial internal carotid artery (V(ICA)) was calculated. Diagnostic accuracy was evaluated by calculating the area under the receiver operating characteristic curve (Az). The significance of the difference between the two Az values (for US vs DSA) was determined by using the z test with correction for correlated data. RESULTS: Nine of 120 patients were excluded because of inadequacy of acoustic windows in the squama of temporal bones. Spasm was mild in 17, moderate in 16, and severe in only nine of 222 arteries studied. Arteries with moderate or severe vasospasm were combined in one group. The best-performing parameters were peak systolic velocity and V(MCA)/V(ICA) ratio. Az values for these two parameters in diagnosis of moderate to-severe vasospasm were 0.93 and 0.95, and in diagnosis of mild vasospasm, 0.90 and 0.91. Accuracy of the V(MCA)/V(ICA) ratio calculated on the basis of end diastolic velocity for diagnosis of mild MCA narrowing was significantly better than that of end-diastolic MCA velocity alone (Az = 0.88 vs 0.84, P < .05). The stepwise approach with use of the V(MCA)/V(ICA) ratio after flow velocity measurements in the MCA resulted in a decreased number of false-negative findings in both vasospasm subgroups. The thresholds of highest efficiency were at a mean velocity of 94 and 108 cm/sec and a peak systolic V(MCA)/V(ICA) ratio of 3.6 and 3.9 for diagnosis of mild and moderate-to-severe vasospasm, respectively. CONCLUSION: Transcranial color-coded duplex US alone or in conjunction with carotid artery US has excellent accuracy for angiographic detection of vasospasm. Use of MCA velocity measurements and V(MCA)/V(ICA) ratio can increase the accuracy of Doppler US. PMID- 16040919 TI - Diffusion-weighted MR imaging in patients with phenylketonuria: relationship between serum phenylalanine levels and ADC values in cerebral white matter. AB - PURPOSE: To prospectively determine the relationship between serum phenylalanine levels and apparent diffusion coefficient (ADC) values in the cerebral white matter of patients with phenylketonuria (PKU). MATERIALS AND METHODS: Institutional review board approval was obtained, and participants provided informed consent. Magnetic resonance (MR) imaging, which included T1- and T2 weighted, fluid-attenuated inversion-recovery (FLAIR), and diffusion-weighted examinations, was performed in 21 patients with PKU (nine male and 12 female patients; age range, 3-44 years; mean age, 19.4 years). ADC values in deep cerebral white matter were calculated for each patient. Serum phenylalanine levels were obtained in all patients within 12 days after MR imaging. Serum phenylalanine levels were measured in 16 patients 1 year before MR imaging. ADC values in cerebral white matter and serum phenylalanine levels were compared. A total of 21 control subjects (12 male and nine female patients; age range, 3-33 years; mean age, 20.6 years) underwent MR imaging. ADC values in cerebral white matter were compared with serum phenylalanine levels by using the Pearson correlation. RESULTS: Abnormal high signal intensity in white matter on T2 weighted and FLAIR MR images was noted in patients with PKU who had serum phenylalanine levels of more than 8.5 mg/dL (514.2 micromol/L). Diffusion in posterior deep cerebral white matter tended to be restricted in patients when increased serum phenylalanine levels were measured after MR imaging (r = -0.62). There was a correlation between ADC values in posterior cerebral white matter and serum phenylalanine levels measured 1 year before MR imaging (r = -0.77). ADCs of control subjects were significantly higher than ADCs of patients with PKU (P < .005). CONCLUSION: Posterior deep white matter in patients with PKU and a serum phenylalanine level of more than 8.5 mg/dL showed high signal intensity in white matter on T2-weighted and FLAIR MR images and revealed decreased ADC. We suggest that to avoid brain-restricted diffusion due to hyperphenylalanemia, patients with PKU should maintain serum phenylalanine levels of less than 8.5 mg/dL (514.2 micromol/L). PMID- 16040920 TI - White matter anisotropy in childhood medulloblastoma survivors: association with neurotoxicity risk factors. AB - PURPOSE: To prospectively evaluate the relationships between change in white matter (WM) anisotropy and (a) patient age at craniospinal irradiation (CSI), (b) CSI dose, and (c) time of magnetic resonance (MR) imaging since CSI and to determine the effect of these neurotoxicity risk factors on WM anisotropy in posttreatment medulloblastoma survivors. MATERIALS AND METHODS: Informed consent was obtained from the patients, control subjects, or their parents, and the study was approved by the institutional review board. Twenty consecutive medulloblastoma survivors (14 male, six female; mean age, 11.0 years +/- 4.6 [standard deviation]) and 36 control subjects (14 male, 22 female; mean age, 10.7 years +/- 3.5) were examined. Control subjects were divided into four groups according to age: 5.0-7.9 years, 8.0-10.9 years, 11.0-13.9 years, and 14.0-18.9 years. The authors calculated the histogram-derived mean WM fractional anisotropy (FA) value for each patient and compared it with the mean WM FA value for the control subjects in the corresponding age group to evaluate the percentage change in WM FA (DeltaFA) in each patient. Spearman rank correlation analysis was used to analyze the relationships between DeltaFA and (a) age at CSI, (b) CSI dose, and (c) time of MR imaging since CSI. Then, multiple linear regression analysis was performed to study the simultaneous influence of these factors on DeltaFA. RESULTS: There were significant correlations between DeltaFA and both age at CSI (r = 0.631, P = .003) and CSI dose (r = -0.586, P = .007) but not between DeltaFA and time of MR imaging since CSI. Multiple linear regression analysis revealed age at CSI to be the only independent variable that significantly affected DeltaFA (adjusted r2= 0.391, P = .012). CONCLUSION: Loss of WM anisotropy is significantly affected by age at CSI, and there is a trend toward increasing anisotropy loss with larger CSI dose. Both age at CSI and CSI dose are known risk factors of neurotoxicity. PMID- 16040921 TI - The calyceal crescent sign. PMID- 16040922 TI - Simultaneous assessment of motor and language areas with a single functional MR imaging paradigm: feasibility. AB - The purpose of this study was to test a hybrid multitask paradigm in healthy subjects and in a patient at preoperative functional magnetic resonance (MR) imaging. In this new paradigm, tasks related to different centers of brain function (motor and language) are performed in alternation during a single functional MR imaging examination, without the typical rest period, to improve time efficiency without compromising the quality of activation maps. The institutional review board approved the study, and all participants gave informed prior consent. The study complied with the Health Insurance Portability and Accountability Act. Seven healthy right-handed volunteers (four men, three women; age range, 22-40 years) were studied first. In both individual and group analyses, there was no statistically significant difference in results between assessment with the multitask design and that with the traditional block design. An advantage of the new paradigm was substantial time savings because the subject was engaged during the entire examination. Finally, the multitask design was tested in a patient undergoing preoperative evaluation, with similar results; therefore, use of the multitask design is feasible in a clinical setting. PMID- 16040923 TI - Intravenous contrast material administration at helical 16-detector row CT coronary angiography: effect of iodine concentration on vascular attenuation. AB - The institutional review board approved this study, and all patients gave written informed consent. One hundred twenty-five patients scheduled to undergo retrospectively electrocardiographically gated 16-detector row computed tomographic coronary angiography were prospectively randomized into the following five groups with respect to the intravenous administration of a 140-mL bolus of contrast material at 4 mL/sec: group 1 (iohexol [300 mg of iodine per milliliter]), group 2 (iodixanol [320 mg I/mL]), group 3 (iohexol [350 mg I/mL]), group 4 (iomeprol [350 mg I/mL]), and group 5 (iomeprol [400 mg I/mL]). Attenuation was measured in the descending aorta and coronary arteries. One-way analysis of variance was used to compare groups. Mean attenuation values in the descending aorta were significantly (P < .05) lower in group 1 and higher in group 5 compared with the mean values in the other three groups. The same pattern was observed in the coronary arteries. Contrast materials with higher iodine concentrations yield significantly higher attenuation in the descending aorta and coronary arteries. PMID- 16040924 TI - Metallic prosthesis: technique to avoid increase in CT radiation dose with automatic tube current modulation in a phantom and patients. AB - The institutional review board approved this Health Insurance Portability and Accountability Act-compliant study protocol, with waiver of informed consent. The purpose of the study was to retrospectively evaluate the combined automatic tube current modulation technique in patients with orthopedic metallic prostheses. Five hundred abdominal-pelvic computed tomographic (CT) studies performed with combined modulation technique were reviewed to identify nine patients with metallic prostheses (mean age, 66 years; range, 35-86 years; male-female ratio, 5:4). On the basis of age and transverse abdominal images, these patients were matched with nine others with no metallic prostheses (mean age, 56 years; range, 36-72 years; male-female ratio, 4:5) who were control patients. Images were graded for extent and severity of streak artifacts (grade 1, streak artifact present but not substantially compromising evaluation of adjacent structures; grade 2, streak artifact present and slightly compromising evaluation of adjacent structures; and grade 3, streak artifact present and severely compromising evaluation of adjacent structures). Student t test was performed for statistical analysis. There was no difference in mean effective tube current-time product between study and control patients (P > .49). With automatic tube current modulation, an increase in CT dose caused by metallic prostheses can be successfully avoided. PMID- 16040925 TI - Pleural disease in silicosis: pleural thickening, effusion, and invagination. AB - PURPOSE: To retrospectively evaluate pleural disease on images from patients with autopsy-proved silicosis. MATERIALS AND METHODS: The study had institutional review board approval, and informed consent from relatives of diseased subjects was waived. Lung specimens were obtained at autopsy in 110 men (mean age, 72 years) who had been followed up radiologically for a mean of 14.8 years. Computed tomographic (CT) scans obtained within 2 years before death were examined for presence of pleural thickening; shape, composition, size, and subpleural location of progressive massive fibrosis (PMF); and pleural invagination (bandlike structure between lesion and pleura). Lung specimens were reviewed and compared with CT findings. Serial chest radiographs and CT scans were reviewed for presence of pleural effusion. Association between radiographic findings and pleural invagination was analyzed with chi2 and Student t tests. Multiple logistic regression analysis was used to find predictive variables for pleural invagination. RESULTS: Pleural effusion was found in 12 (11%) patients at chest radiography and CT, and thickening was found in 64 (58%) patients at CT; the latter finding was significantly more frequent with complicated silicosis (P < .001). At CT, 128 PMF lesions were seen, 39 (30%) of which showed pleural invagination; CT scans showed pleural thickening in 36 (92%) of these 39 lesions. In 17 (44%) PMF lesions, CT scans depicted a bandlike structure that was pathologically confirmed to represent invaginated pleura in all cases. Pathologic presence of invagination was significantly associated with pleural thickening (P < .001), ipsilateral pleural effusion (P < .01), interstitial fibrosis (P < .05), and the nearness of PMF to the pleura (P < .005). Multiple logistic regression analysis showed that pleural thickening (odds ratio, 62.51; 95% confidence interval [CI]: 5.564, 70.2) and pleural effusion (odds ratio, 25.865; 95% CI: 1.992, 335.8) were significant CT variables associated with presence of pathologic pleural invagination (P = .001 and .013, respectively). Five PMF lesions had radiographic features of rounded atelectasis. CONCLUSION: Various pleural abnormalities can occur in silicosis, especially in advanced disease. PMID- 16040926 TI - Evaluation of bowel ischemia with contrast-enhanced US: initial experience. AB - PURPOSE: To prospectively evaluate the accuracy of contrast material-enhanced ultrasonography (US) in the depiction of bowel ischemia in patients with radiographic evidence of small-bowel dilatation. MATERIALS AND METHODS: The ethics committee approved this study, and informed consent was obtained from all patients. Fifty-one patients (34 men and 17 women; mean age, 67.1 years) with evidence of small-bowel dilatation at conventional radiography were enrolled. Twenty patients had bowel ischemia (15 cases of bowel strangulation and five of thromboembolism of the superior mesenteric artery) and 31 patients had simple obstruction. After injection of SHU-508A, the most dilated or the least peristaltic bowel segments were imaged at contrast-enhanced power Doppler US (interval, 4 seconds) for 2 minutes. Color signals obtained in the bowel wall were classified as normal, diminished, or absent. Contrast-enhanced US and classification of color signals were performed by a sonologist. The US systems were equipped with 3-12-MHz transducers. Fisher exact test was used to evaluate the significance of the differences between each group of patients, and P < .01 was considered to indicate a significant difference. RESULTS: The color signals were absent in five patients with superior mesenteric arterial thromboembolism and in seven patients with strangulation, were diminished in five patients with strangulation, and were normal in three patients with strangulation and in 31 patients with simple obstruction. By pooling the absent and diminished color signals together as a diagnostic indicator of bowel ischemia, the sensitivity was 85% (95% confidence interval [CI]: 62.1%, 96.8%), the specificity was 100% (95% CI: 90.8%, 100%), the positive predictive value was 100% (95% CI: 83.8%, 100%), the negative predictive value was 91.2% (95% CI: 76.3%, 98.1%), the likelihood ratio for a positive test result was infinity, and the likelihood ratio for a negative test result was 0.15 (95% CI: 0.032, 0.379). CONCLUSION: Contrast enhanced US shows promise for the noninvasive diagnosis of bowel ischemia based on initial experience in patients with radiographic evidence of small-bowel dilatation. PMID- 16040927 TI - Renal tumors: MR imaging-guided percutaneous cryotherapy--initial experience in 23 patients. AB - PURPOSE: To evaluate the initial clinical experience of magnetic resonance (MR) imaging-guided percutaneous cryotherapy of renal tumors. MATERIALS AND METHODS: Twenty-six renal tumors (diameter range, 1.0-4.6 cm; mean, 2.6 cm) in 23 patients were treated with 27 cryoablation procedures by using a protocol approved by the human subjects committee at the authors' institution. The study complied with the Health Insurance Portability and Accountability Act. Written informed consent was obtained from each patient. There were 17 men and six women with an average age of 66 years (range, 43-86 years). Of 26 masses, 24 were renal cell carcinoma, one was a transitional cell carcinoma, and one was an angiomyolipoma. By using a 0.5 T open MR imaging system and general anesthesia in patients, one to five (mean, 2.4) needlelike cryoprobes were placed and lesions were ablated by using real time MR imaging for intraprocedural monitoring of ice balls. Tumors were considered successfully ablated if they demonstrated no contrast enhancement at follow-up computed tomography or MR imaging (mean, 14 months; range, 4-30 months). RESULTS: Twenty-four of 26 tumors were successfully ablated, 23 of which required only one treatment session. Two complications occurred in a total of 27 cryoablations: one hemorrhage, which required a blood transfusion, and one abscess, which was treated successfully with percutaneous catheter drainage. CONCLUSION: MR imaging-guided percutaneous cryotherapy of renal tumors shows promise for the treatment of selected small renal tumors, and MR imaging can be used to monitor the treatment intraprocedurally. This technique may prove useful for ablation of renal tumors completely in one session, but long-term follow-up is needed. PMID- 16040928 TI - Fungus-infected fluid collections in thorax or abdomen: effectiveness of percutaneous catheter drainage. AB - PURPOSE: To retrospectively evaluate the effectiveness of percutaneous catheter drainage in the treatment of fungus-infected fluid collections in the thorax or abdomen and to identify any factor that may be predictive of a poor clinical outcome. MATERIALS AND METHODS: Approval for this study was obtained from the hospital ethics subcommittee on human studies. Because the study was retrospective, patient informed consent was not required. This study was compliant with the Health Insurance Portability and Accountability Act. Retrospective analysis was performed of cases of fungus-infected fluid collections in the thorax or abdomen treated by using percutaneous catheter drainage in 60 patients (36 male and 24 female patients; mean age, 57 years; range, 2 months to 91 years) during 5 years. The patient medical records were reviewed to identify recognized factors for predisposition to fungal infection. The details of percutaneous catheter drainage and microbiologic findings were recorded. The technical success (ability of catheters placed to drain collections treated) and the clinical success (ability of patients to recover fully without surgery) of percutaneous catheter drainage were determined. A multifactor logistic regression analysis was performed to identify any clinical or microbiologic factor predictive of a poor clinical outcome. RESULTS: Seventy three fungus-infected fluid collections were drained in 60 patients. The collections originated from the pleura (n = 6), mediastinum (n = 2), liver (n = 3), pancreas (n = 5), obstructed biliary or urinary tract (n = 9), gallbladder (n = 1), and abdominopelvic area (n = 47). The technical success rate for catheter drainage was 79% (41 of 52 patients); the clinical success rate, 57% (34 of 60 patients). Twenty (33%) patients died from all causes during hospital admission. Multifactor logistic regression analysis was used to identify predictors of a poor clinical outcome; complexity of collection, history of malignancy, and admission to intensive care unit were significant (P < .03) and independent predictors. CONCLUSION: Despite a moderately high technical success rate with percutaneous catheter drainage of fungus-infected fluid collections, clinical success rate was much lower. Both imaging appearance (complexity of collection) and clinical factors (history of malignancy, admission to intensive care unit) influenced prognosis. PMID- 16040929 TI - Molecular markers, paraneoplastic syndrome, and inferior vena cava involvement as risk factors for renal tumor recurrence. PMID- 16040930 TI - Comparison of moderate versus high concentration of contrast media injected at the same total iodine dose and fixed injection duration. PMID- 16040932 TI - Abstracts of the 49th Annual Scientific Meeting of the Society for Social Medicine. September 14-16, 2005. Glasgow, United Kingdom. PMID- 16040938 TI - Effects of OP exposure in people. PMID- 16040941 TI - Frequency and cost of human intervention at lambing: an interbreed comparison. AB - The frequency of human intervention during lambing was assessed for 708 lambs and 162 ewes in up to four deliveries. In total, 21.8 per cent of the lambs born alive required some assistance at birth and 9.4 per cent were delivered manually. Assistance was required less frequently by twin lambs and ewe lambs, and Scottish blackface lambs required assistance less frequently than Suffolk lambs. Approximately one-third of the ewes were assisted in at least one delivery and 10 per cent were assisted in all their deliveries; however, less than 20 per cent of ewes were assisted more than once. A quarter of the lambs needed some human intervention to suck successfully, and Suffolk lambs needed help more frequently. The lambs born to almost half the first-parity ewes needed help to suck, but at later deliveries the lambs born to less than a third of the ewes needed assistance. In blackface flocks the survival of 100 lambs required 4.6 hours of human intervention whereas in Suffolk flocks 28 hours were required. PMID- 16040942 TI - Evaluation of the aerosol transmission of a mixed infection of Mycoplasma hyopneumoniae and porcine reproductive and respiratory syndrome virus. AB - To evaluate the transmission of Mycoplasma hyopneumoniae and porcine reproductive and respiratory syndrome virus (PRRSV) by aerosol as either a single or mixed infection, 28 pigs were inoculated intratracheally with M hyopneumoniae on day 0 and infected intranasally with PRRSV on day 35; they were housed together in a barn. To assess the aerosol transmission of M hyopneumoniae as a single infection, one trailer (A) containing 10 five-week-old sentinel pigs was placed along the south side of the infected barn (1 m from the fans) on day 28. To assess the mixed infection, two trailers (B and C), each containing 10 five-week old sentinel pigs, were placed along each side of the barn on day 42. The sentinel pigs in the three trailers were exposed to the exhaust from the fans for seven days. No M hyopneumoniae infection was detected in the sentinel pigs in trailer A, but it was detected in the sentinel pigs in trailers B and C. No PRRSV was detected in any of the sentinel pigs. PMID- 16040943 TI - Plasma 14C-cholic acid clearance in healthy dogs and dogs with cholestasis or a congenital portosystemic shunt. AB - The clearance profiles of intravenously injected tracer doses of radioactively labelled cholic acid were investigated in healthy dogs, dogs with a congenital portosystemic shunt and dogs with cholestasis. The rate constants and residual plasma activity were significantly different in the healthy and diseased dogs, but it was not possible to differentiate between the dogs with portosystemic shunting and cholestasis because the results were determined not only by factors involved in plasma bile acid clearance but also by the enterohepatic circulation. PMID- 16040944 TI - Outbreaks of classical swine fever in the Republic of Korea in 2003. PMID- 16040945 TI - Clinical, pathological and diagnostic aspects of congenital neosporosis in a series of naturally infected calves. PMID- 16040946 TI - Enzootic intranasal tumour virus infection in apparently healthy sheep in Japan. PMID- 16040947 TI - Serological surveillance of canine leptospirosis in Mongolia. PMID- 16040948 TI - Intestinal dilation in layers. PMID- 16040950 TI - Control of nematode infections in horses. PMID- 16040954 TI - Uterine torsion in cattle in the UK. PMID- 16040956 TI - TcpF is a soluble colonization factor and protective antigen secreted by El Tor and classical O1 and O139 Vibrio cholerae serogroups. AB - Vibrio cholerae causes diarrhea by colonizing the human small bowel and intoxicating epithelial cells. Colonization is a required step in pathogenesis, and strains defective for colonization are significantly attenuated. The best characterized V. cholerae colonization factor is the toxin-coregulated pilus (TCP). It has been demonstrated that TCP is required for V. cholerae colonization in both humans and mice. TCP enhances bacterial interactions that allow microcolony formation and thereby promotes survival in the intestine. We have recently discovered that the TCP biogenesis apparatus also serves as a secretion system, mediating the terminal step in the extracellular secretion pathway of TcpF. TcpF was identified in classical isolates of V. cholerae O1 as a soluble factor essential for colonization in the infant mouse cholera model. In the present study, we expanded our analysis of TcpF to include the O1 El Tor and O139 serogroups and investigated how TCP and TcpF act together to mediate colonization. Additionally, we demonstrated that antibodies generated against TcpF are protective against experimental V. cholerae infection in the infant mouse cholera model. This observation, coupled with the fact that TcpF is a potent mediator of colonization, suggests that TcpF should be considered as a component of a polyvalent cholera vaccine formulation. PMID- 16040957 TI - An ABC transporter containing a forkhead-associated domain interacts with a serine-threonine protein kinase and is required for growth of Mycobacterium tuberculosis in mice. AB - Forkhead-associated (FHA) domains are modular phosphopeptide recognition motifs with a striking preference for phosphothreonine-containing epitopes. FHA domains have been best characterized in eukaryotic signaling pathways but have been identified in six proteins in Mycobacterium tuberculosis, the causative organism of tuberculosis. One of these, coded by gene Rv1747, is an ABC transporter and the only one to contain two such modules. A deletion mutant of Rv1747 is attenuated in a mouse intravenous injection model of tuberculosis where the bacterial load of the mutant is 10-fold lower than that of the wild type in both lungs and spleen. In addition, growth of the mutant in mouse bone marrow-derived macrophages and dendritic cells is significantly impaired. In contrast, growth of this mutant in vitro was indistinguishable from that of the wild type. The mutant phenotype was lost when the mutation was complemented by the wild-type allele, confirming that it was due to mutation of Rv1747. Using yeast two-hybrid analysis, we have shown that the Rv1747 protein interacts with the serine threonine protein kinase PknF. This interaction appears to be phospho-dependent since it is abrogated in a kinase-dead mutant and by mutations in the presumed activation loop of PknF and in the first FHA domain of Rv1747. These results demonstrate that the protein coded by Rv1747 is required for normal virulent infection by M. tuberculosis in mice and, since it interacts with a serine threonine protein kinase in a kinase-dependent manner, indicate that it forms part of an important phospho-dependent signaling pathway. PMID- 16040958 TI - N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 Inhibitor with Salmonella enterica serovar typhimurium lipopolysaccharide and lipid A. AB - The C1 inhibitor (C1INH), a plasma complement regulatory protein, prevents endotoxin shock, at least partially via the direct interaction of its amino terminal heavily glycosylated nonserpin region with gram-negative bacterial lipopolysaccharide (LPS). To further characterize the potential LPS-binding site(s) within the amino-terminal domain, mutations were introduced into C1INH at the three N-linked glycosylation sites and at the four positively charged amino acid residues. A mutant in which Asn(3) was replaced with Ala was markedly less effective in its binding to LPS, while substitution of Asn(47) or Asn(59) had little effect on binding. The mutation of C1INH at all four positively charged amino acid residues (Arg(18), Lys(22), Lys(30), and Lys(55)) resulted in near complete failure to interact with LPS. The C1INH mutants that did not bind to LPS also did not suppress LPS binding or LPS-induced up-regulation of tumor necrosis factor alpha mRNA expression in RAW 264.7 macrophages. In addition, the binding of C1INH mutants to diphosphoryl lipid A was decreased in comparison with that of recombinant wild-type C1INH. Therefore, the interaction of C1INH with gram negative bacterial LPS is dependent both on the N-linked carbohydrate at Asn(3) and on the positively charged residues within the amino-terminal domain. PMID- 16040959 TI - Transcriptional profiling of Vibrio cholerae recovered directly from patient specimens during early and late stages of human infection. AB - Understanding gene expression by bacteria during the actual course of human infection may provide important insights into microbial pathogenesis. In this study, we evaluated the transcriptional profile of Vibrio cholerae, the causative agent of cholera, in clinical specimens from cholera patients. We collected samples of human stool and vomitus that were positive by dark-field microscopy for abundant vibrios and used a microarray to compare gene expression in organisms recovered directly from specimens collected during the early and late stages of human infection. Our results reveal that V. cholerae gene expression within the human host environment differs from patterns defined in in vitro models of pathogenesis. tcpA, the major subunit of the essential V. cholerae colonization factor, was significantly more highly expressed in early than in late stages of infection; however, the genes encoding cholera toxin were not highly expressed in either phase of human infection. Furthermore, expression of the virulence regulators toxRS and tcpPH was uncoupled. Interestingly, the pattern of gene expression indicates that the human upper intestine may be a uniquely suitable environment for the transfer of genetic elements that are important in the evolution of pathogenic strains of V. cholerae. These findings provide a more detailed assessment of the transcriptome of V. cholerae in the human host than previous studies of organisms in stool alone and have implications for cholera control and the design of improved vaccines. PMID- 16040960 TI - Specificity of Legionella pneumophila and Coxiella burnetii vacuoles and versatility of Legionella pneumophila revealed by coinfection. AB - Legionella pneumophila and Coxiella burnetii are phylogenetically related intracellular bacteria that cause aerosol-transmitted lung infections. In host cells both pathogens proliferate in vacuoles whose biogenesis displays some common features. To test the functional similarity of their respective intracellular niches, African green monkey kidney epithelial (Vero) cells, A/J mouse bone marrow-derived macrophages, human macrophages, and human dendritic cells (DC) containing mature C. burnetii replication vacuoles were superinfected with L. pneumophila, and then the acidity, lysosome-associated membrane protein (LAMP) content, and cohabitation of mature replication vacuoles was assessed. In all cell types, wild-type L. pneumophila occupied distinct vacuoles in close association with acidic, LAMP-positive C. burnetii replication vacuoles. In murine macrophages, but not primate macrophages, DC, or epithelial cells, L. pneumophila replication vacuoles were acidic and LAMP positive. Unlike wild-type L. pneumophila, type IV secretion-deficient dotA mutants trafficked to lysosome like C. burnetii vacuoles in Vero cells where they survived but failed to replicate. In primate macrophages, DC, or epithelial cells, growth of L. pneumophila was as robust in superinfected cell cultures as in those singly infected. Thus, despite their noted similarities, L. pneumophila and C. burnetii are exquisitely adapted for replication in unique replication vacuoles, and factors that maintain the C. burnetii replication vacuole do not alter biogenesis of an adjacent L. pneumophila replication vacuole. Moreover, L. pneumophila can replicate efficiently in either lysosomal vacuoles of A/J mouse cells or in nonlysosomal vacuoles of primate cells. PMID- 16040961 TI - Human {beta}-defensin 2 is expressed and associated with Mycobacterium tuberculosis during infection of human alveolar epithelial cells. AB - To determine the role of human beta-defensin 2 (HBD-2) in human tuberculosis, we studied the in vitro induction of HBD-2 gene expression by Mycobacterium tuberculosis H37Rv infection in the human lung epithelial cell line A549, in alveolar macrophages (AM), and in blood monocytes (MN) by reverse transcription PCR. We also studied the induction of HBD-2 gene expression by mannose lipoarabinomannan (manLAM) from M. tuberculosis. Intracellular production of HBD 2 peptide was detected by immunocytochemistry and electron microscopy. Our results demonstrated that there was induction of HBD-2 mRNA in A549 cells after infection with M. tuberculosis at various multiplicities of infection (MOI) and that there was stimulation with manLAM. AM expressed the HBD-2 gene only at a high MOI with M. tuberculosis. MN did not express HBD-2 at any of the experimental M. tuberculosis MOI. Immunostaining revealed the presence of intracellular HBD-2 peptide in A549 cells following infection with M. tuberculosis, and the staining was more intense in areas where there were M. tuberculosis clusters. By using electron microscopy we also demonstrated production of HBD-2 after M. tuberculosis infection and adherence of HBD-2 to the membranes of M. tuberculosis. Alveolar epithelial cells are among the first cells to encounter M. tuberculosis following aerogenic infection. As HBD-2 has been shown to control growth of M. tuberculosis and has chemotactic activity, our results suggest that HBD-2 induction by M. tuberculosis may have a role in the pathogenesis of human tuberculosis. PMID- 16040962 TI - A toll-like receptor 2-responsive lipid effector pathway protects mammals against skin infections with gram-positive bacteria. AB - flake (flk), an N-ethyl-N-nitrosourea-induced recessive germ line mutation of C57BL/6 mice, impairs the clearance of skin infections by Streptococcus pyogenes and Staphylococcus aureus, gram-positive pathogens that elicit innate immune responses by activating Toll-like receptor 2 (TLR2). Positional cloning and sequencing revealed that flk is a novel allele of the stearoyl coenzyme A desaturase 1 gene (Scd1). flake homozygotes show reduced sebum production and are unable to synthesize the monounsaturated fatty acids (MUFA) palmitoleate (C(16:1)) and oleate (C(18:1)), both of which are bactericidal against gram positive (but not gram-negative) organisms in vitro. However, intradermal MUFA administration to S. aureus-infected mice partially rescues the flake phenotype, which indicates that an additional component of the sebum may be required to improve bacterial clearance. In normal mice, transcription of Scd1-a gene with numerous NF-kappaB elements in its promoter--is strongly and specifically induced by TLR2 signaling. Similarly, the SCD1 gene is induced by TLR2 signaling in a human sebocyte cell line. These observations reveal the existence of a regulated, lipid-based antimicrobial effector pathway in mammals and suggest new approaches to the treatment or prevention of infections with gram-positive bacteria. PMID- 16040963 TI - Resistance to intestinal Entamoeba histolytica infection is conferred by innate immunity and Gr-1+ cells. AB - Establishment of intestinal infection with Entamoeba histolytica depends on the mouse strain; C57BL/6 mice are highly resistant, and C3H/HeJ mice are relatively susceptible. We found that resistance to intestinal infection was independent of lymphocyte activity or H-2 haplotype and occurred in the first hours to days postchallenge according to in vivo imaging. At 18 h postchallenge, the ceca of resistant C57BL/6 mice were histologically unremarkable, in contrast to the severe inflammation observed in susceptible C3H/HeJ mice. Comparison of cecal gene expression in C3H/HeJ and C57BL/6 mice demonstrated that there was parasite induced upregulation of proinflammatory and neutrophil chemotaxis transcripts and there was downregulation of transforming growth factor beta signaling molecules. Pretreatment with dexamethasone abrogated the partial resistance of C3H/HeJ or CBA mice through an innate, lymphocyte-independent mechanism, but it had no effect on the high-level resistance of C57BL/6 mice. Similarly, administration of a neutrophil-depleting anti-Gr-1 monoclonal antibody (RB6-8C5) decreased the partial resistance of CBA mice and led to severe pathology compared to control antibody-treated mice, but it had no effect on C57BL/6 resistance. These data indicate that there are discrete mechanisms of innate resistance to E. histolytica depending on the host background and, in contrast to other reports, imply that neutrophils are protective and not damaging in intestinal amebiasis. PMID- 16040964 TI - Modulation of the lung inflammatory response to serotype 8 pneumococcal infection by a human immunoglobulin m monoclonal antibody to serotype 8 capsular polysaccharide. AB - The human monoclonal antibody to serotype 8 pneumococcal capsular polysaccharide D11 [immunoglobulin M(kappa)] protects wild-type and complement component 4 knockout (C4 KO) mice against lethal intratracheal challenge with serotype 8 pneumococcus, but it does not promote polymorphonuclear leukocyte (PMN)-mediated pneumococcal killing in vitro. In this study, we investigated the effect of D11 on the blood and lung bacterial burdens and the serum and lung expression of inflammatory chemokines and cytokines in an intratracheal challenge model with serotype 8 pneumococcus in C4 KO mice. Pneumococcus was not detected in the blood of D11-treated mice, whereas control mice had high-grade bacteremia with >10(7) CFU. Control mice had a >5-log increase in lung CFU and D11-treated mice manifested a nearly 3-log increase in lung CFU compared to the original inoculum 24 h after infection. Serum and lung levels of soluble macrophage inflammatory protein 2 (MIP-2) and interleulin-6 (IL-6) as measured by an enzyme-linked immunosorbent assay were lower in D11-treated mice than in control mice 24 h after infection. Real-time PCR was performed to examine lung mRNA chemokine and cytokine expression. The results showed that D11-treated mice had significantly less gamma interferon, MIP-2, IL-12, monocyte chemoattractant protein 1/JE, and tumor necrosis factor alpha expression than control mice 24 h after infection. Histopathology and immunohistochemical staining of lung tissues revealed that D11 treated mice had less inflammation, fewer PMNs, and less myeloperoxidase staining than control mice 24 h after infection. These findings suggest that the efficacy of certain serotype-specific antibodies against pneumococcal pneumonia could be associated with modulation of the lung inflammatory response and a reduction in host damage. PMID- 16040965 TI - Stimulation of gp91 phagocytic oxidase and reactive oxygen species in neutrophils by an avirulent Salmonella enterica serovar infantis strain protects gnotobiotic piglets from lethal challenge with serovar Typhimurium strain F98 without inducing intestinal pathology. AB - Preinoculation of susceptible 5-day-old gnotobiotic piglets with Salmonella enterica serovar Infantis strain 1326/28Phi(r) stimulates neutrophil migration into the intestine, which rapidly protects the pigs against a subsequent (normally lethal) challenge with S. enterica serovar Typhimurium strain F98. Here we show that inoculation with either 1326/28Phi(r) or F98 activated reactive oxygen species (ROS) in neutrophils via NADPH pathways in vivo and in vitro and that the survival of both Salmonella strains was increased if neutrophils were cocultured with the ROS inhibitor N-acetylcysteine (captopril). Neither F98 nor 1326/28Phi(r) significantly increased reactive nitrogen species (RNS) levels in neutrophils isolated from uninfected pigs. Our results indicate the following: (i) rapid protection of highly susceptible gnotobiotic piglets against F98 induced gastroenteritis by preinoculation with 1326/28Phi(r) is likely to be due to stimulation of ROS-producing neutrophils in the intestinal epithelium prior to challenge with the lethal strain; (ii) pathological lesions of the intestine during severe gastroenteritis are not necessarily induced by neutrophil migration per se; and (iii) if neutrophil migration into the intestine is responsible for pathology, then neither increased production of ROS or RNS (in pigs inoculated with the lethal strain) nor reduced production (in protected pigs in which pathological lesions are ameliorated by preinoculation with 1326/28Phi(r)) can account for this phenomenon. PMID- 16040966 TI - Aspergillus cyclooxygenase-like enzymes are associated with prostaglandin production and virulence. AB - Oxylipins comprise a family of oxygenated fatty acid-derived signaling molecules that initiate critical biological activities in animals, plants, and fungi. Mammalian oxylipins, including the prostaglandins (PGs), mediate many immune and inflammation responses in animals. PG production by pathogenic microbes is theorized to play a role in pathogenesis. We have genetically characterized three Aspergillus genes, ppoA, ppoB, and ppoC, encoding fatty acid oxygenases similar in sequence to specific mammalian prostaglandin synthases, the cyclooxygenases. Enzyme-linked immunosorbent assay analysis showed that production of PG species is decreased in both Aspergillus nidulans and A. fumigatus ppo mutants, implicating Ppo activity in generating PGs. The A. fumigatus triple-ppo-silenced mutant was hypervirulent in the invasive pulmonary aspergillosis murine model system and showed increased tolerance to H(2)O(2) stress relative to that of the wild type. We propose that Ppo products, PG, and/or other oxylipins may serve as activators of mammalian immune responses contributing to enhanced resistance to opportunistic fungi and as factors that modulate fungal development contributing to resistance to host defenses. PMID- 16040967 TI - Degradation of Chlamydia pneumoniae by peripheral blood monocytic cells. AB - Chlamydia pneumoniae is a common human respiratory pathogen that has been associated with a variety of chronic diseases, including atherosclerosis. The role of this organism in the pathogenesis of atherosclerosis remains unknown. A key question is how C. pneumoniae is transferred from the site of primary infection to a developing atherosclerotic plaque. It has been suggested that circulating monocytes could be vehicles for dissemination of C. pneumoniae since the organism has been detected in peripheral blood monocytic cells (PBMCs). In this study we focused on survival of C. pneumoniae within PBMCs isolated from the blood of healthy human donors. We found that C. pneumoniae does not grow and multiply in cultured primary monocytes. In C. pneumoniae-infected monocyte derived macrophages, growth of the organism was very limited, and the majority of the bacteria were eradicated. We also found that the destruction of C. pneumoniae within infected macrophages resulted in a gradual diminution of chlamydial antigens, although some of these antigens could be detected for days after the initial infection. The detected antigens present in infected monocytes and monocyte-derived macrophages represented neither chlamydial inclusions nor intact organisms. The use of {N-[7-(4-nitrobenzo-2-oxa-1,3-diazole)]}-6-aminocaproyl-d erythro-sphingosine as a vital stain for chlamydiae proved to be a sensitive method for identifying rare C. pneumoniae inclusions and was useful in the detection of even aberrant developmental forms. PMID- 16040968 TI - Virulence of the fungal pathogen Candida albicans requires the five isoforms of protein mannosyltransferases. AB - The PMT gene family in Candida albicans encodes five isoforms of protein mannosyltransferases (Pmt proteins Pmt1p, Pmt2p, Pmt4p, Pmt5p, and Pmt6p) that initiate O mannosylation of secretory proteins. We compared virulence characteristics of pmt mutants in two complex, three-dimensional models of localized candidiasis, using reconstituted human epithelium (RHE) and engineered human oral mucosa (EHOM); in addition, mutants were tested in a mouse model of hematogenously disseminated candidiasis (HDC). All pmt mutants showed attenuated virulence in the HDC model and at least one model of localized candidiasis. The pmt5 mutant, which lacks in vitro growth phenotypes, was less virulent in the EHOM and HDC assays but had no consistent phenotype in the RHE assay. In contrast, the pmt4 and pmt6 mutants were less virulent in the RHE and HDC assays but not in the EHOM assay. The results stress the contribution of all Pmt isoforms to the virulence of C. albicans and suggest that the importance of individual Pmt isoforms may differ in specific host niches. We propose that Pmt proteins may be suitable targets for future novel classes of antifungal agents. PMID- 16040969 TI - Role for nucleotide excision repair in virulence of Mycobacterium tuberculosis. AB - Mutations in Mycobacterium tuberculosis uvrB result in severe sensitivity to acidified nitrite, a source of nitric oxide (6). In this study, we show that a uvrB mutant is exquisitely sensitive to UV light but not to several sources of reactive oxygen species in vitro. Furthermore, a uvrB mutant was attenuated in mice as judged by an extension of life span. Attenuation in mice was partially reversed by genetic inactivation of nitric oxide synthase 2 (iNOS) and almost completely reversed in mice lacking both iNOS and phagocyte oxidase. Thus, a gene predicted to encode a key element of DNA repair is required for resistance of M. tuberculosis to both reactive nitrogen and reactive oxygen species in mice. PMID- 16040970 TI - Invasive phenotype of Candida albicans affects the host proinflammatory response to infection. AB - Candida albicans is a major opportunistic pathogen in immunocompromised patients. Production of proinflammatory cytokines by host cells in response to C. albicans plays a critical role in the activation of immune cells and final clearance of the organism. Invasion of host cells and tissues is considered one of the virulence attributes of this organism. The purpose of this study was to investigate whether the ability of C. albicans to invade host cells and tissues affects the proinflammatory cytokine responses by epithelial and endothelial cells. In this study we used the invasion-deficient RIM101 gene knockout strain DAY25, the highly invasive strain SC5314, and highly invasive RIM101-complemented strain DAY44 to compare the proinflammatory cytokine responses by oral epithelial or endothelial cells. Using a high-throughput approach, we found both qualitative and quantitative differences in the overall inflammatory responses to C. albicans strains with different invasive potentials. Overall, the highly invasive strains triggered higher levels of proinflammatory cytokines in host cells than the invasion-deficient mutant triggered. Significant differences compared to the attenuated mutant were noted in interleukin-1alpha (IL-1alpha), IL-6, IL-8, and tumor necrosis factor alpha in epithelial cells and in IL-6, growth-related oncogene, IL-8, monocyte chemoattractant protein 1 (MCP-1), MCP-2, and granulocyte colony-stimulating factor in endothelial cells. Our results indicate that invasion of host cells and tissues by C. albicans enhances the host proinflammatory response to infection. PMID- 16040971 TI - Heparin stimulates Staphylococcus aureus biofilm formation. AB - Heparin, known for its anticoagulant activity, is commonly used in catheter locks. Staphylococcus aureus, a versatile human and animal pathogen, is commonly associated with catheter-related bloodstream infections and has evolved a number of mechanisms through which it adheres to biotic and abiotic surfaces. We demonstrate that heparin increased biofilm formation by several S. aureus strains. Surface coverage and the kinetics of biofilm formation were stimulated, but primary attachment to the surface was not affected. Heparin increased S. aureus cell-cell interactions in a protein synthesis-dependent manner. The addition of heparin rescued biofilm formation of hla, ica, and sarA mutants. Our data further suggest that heparin stimulation of biofilm formation occurs neither through an increase in sigB activity nor through an increase in polysaccharide intracellular adhesin levels. These finding suggests that heparin stimulates S. aureus biofilm formation via a novel pathway. PMID- 16040972 TI - Human antibody against shiga toxin 2 administered to piglets after the onset of diarrhea due to Escherichia coli O157:H7 prevents fatal systemic complications. AB - Infection of children with Shiga toxin (Stx)-producing Escherichia coli (STEC) can lead to hemolytic-uremic syndrome (HUS) in 5 to 10% of patients. Stx2, one of two toxins liberated by the bacterium, is directly linked with HUS. We have previously shown that Stx-specific human monoclonal antibodies protect STEC infected animals from fatal systemic complications. The present study defines the protective antibody dose in relation to the time of treatment after the onset of diarrhea in infected gnotobiotic piglets. Using the mouse toxicity model, we selected 5C12, an antibody specific for the A subunit, as the most effective Stx2 antibody for further characterization in the piglet model in which piglets developed diarrhea 16 to 40 h after bacterial challenge, followed by fatal neurological symptoms at 48 to 96 h. Seven groups of piglets received doses of 5C12 ranging from 6.0 mg/kg to 0.05 mg/kg of body weight, administered parenterally 48 h after bacterial challenge. The minimum fully protective antibody dose was 0.4 mg/kg, and the corresponding serum antibody concentration in these piglets was 0.7 mug (+/-0.5)/ml, measured 7 to 14 days after administration. Of 40 infected animals which received Stx2 antibody treatment of > or =0.4 mg/kg, 34 (85%) survived, while only 1 (2.5%) of 39 placebo-treated animals survived. We conclude that the administration of the Stx2-specific antibody was protective against fatal systemic complications even when it was administered well after the onset of diarrhea. These findings suggest that children treated with this antibody, even after the onset of bloody diarrhea, may be equally protected against the risk of developing HUS. PMID- 16040973 TI - Deletion of the anaerobic regulator HlyX causes reduced colonization and persistence of Actinobacillus pleuropneumoniae in the porcine respiratory tract. AB - Actinobacillus pleuropneumoniae, the etiological agent of porcine pleuropneumonia, is able to persist on respiratory epithelia, in tonsils, and in the anaerobic environment of encapsulated lung sequesters. We have demonstrated previously that putative HlyX-regulated genes, coding for dimethyl sulfoxide (DMSO) reductase and aspartate ammonia lyase, are upregulated during infection and that deletions in these genes result in attenuation of the organism. The study presented here investigates the role of HlyX, the fumarate nitrate reductase regulator (FNR) homologue of A. pleuropneumoniae. By constructing an isogenic A. pleuropneumoniae hlyX mutant, the HlyX protein is shown to be responsible for upregulated expression of both DMSO reductase and aspartate ammonia lyase (AspA) under anaerobic conditions. In a challenge experiment the A. pleuropneumoniae hlyX mutant is shown to be highly attenuated, unable to persist in healthy lung epithelium and tonsils, and impaired in survival inside sequestered lung tissue. Further, using an A. pleuropneumoniae strain carrying the luxAB genes as transcriptional fusion to aspA on the chromosome, the airway antioxidant glutathione was identified as one factor potentially responsible for inducing HlyX-dependent gene expression of A. pleuropneumoniae in epithelial lining fluid. PMID- 16040974 TI - Chlamydia pneumoniae uses the mannose 6-phosphate/insulin-like growth factor 2 receptor for infection of endothelial cells. AB - Several mechanisms for attachment and entry of Chlamydia have been proposed. We previously determined that the major outer membrane protein of Chlamydia trachomatis is glycosylated with a high-mannose oligosaccharide, and a similar structure inhibited the attachment and infectivity of C. trachomatis in epithelial cells. Because insulin-like growth factor 2 (IGF2) was shown to enhance the infectivity of Chlamydia pneumoniae but not C. trachomatis in endothelial cells, a hapten inhibition assay was used to analyze whether the mannose 6-phosphate (M6P)/IGF2 receptor that also binds M6P could be involved in infection of endothelial cells (HMEC-1) by Chlamydia. M6P and mannose 6-phosphate poly[N-(2-hydroxyethyl)-acrylamide] (M6P-PAA) inhibited the infectivity of C. pneumoniae AR-39, but not C. trachomatis serovar UW5 or L2, while mannan inhibited the growth of C. trachomatis, but not C. pneumoniae. Using metabolically labeled organisms incubated with cells at 4 degrees C (organisms attach but do not enter) or at 37 degrees C (organisms attach and are internalized), M6P-PAA was shown to inhibit attachment and internalization of C. pneumoniae in endothelial cells but did not inhibit attachment or internalization of C. trachomatis serovar E or L2. These findings indicate that C. pneumoniae can utilize the M6P/IGF2 receptor and that the use of this receptor for attachment and entry differs between C. pneumoniae and C. trachomatis. PMID- 16040975 TI - Capsule and fimbria interaction in Klebsiella pneumoniae. AB - The capsular polysaccharide and type 1 fimbriae are two of the major surface located virulence properties associated with the pathogenesis of Klebsiella pneumoniae. The capsule is an elaborate polysaccharide matrix that encases the entire cell surface and provides resistance against many host defense mechanisms. In contrast, type 1 fimbriae are thin adhesive thread-like surface organelles that can extend beyond the capsular matrix and mediate d-mannose-sensitive adhesion to host epithelial cells. These fimbriae are archetypical and consist of a major building block protein (FimA) that comprises the bulk of the organelle and a tip-located adhesin (FimH). It is assumed that the extended major-subunit protein structure permits the FimH adhesin to function independently of the presence of a capsule. In this study, we have employed a defined set of K. pneumoniae capsulated and noncapsulated strains to show that the function of type 1 fimbriae is actually impeded by the concomitant expression of a polysaccharide capsule. Capsule expression had significant effects on two parameters commonly used to define FimH function, namely, yeast cell agglutination and biofilm formation. Our data suggest that this effect is not due to transcriptional/translational changes in fimbrial gene/protein expression but rather the result of direct physical interference. This was further demonstrated by the fact that we could restore fimbrial function by inhibiting capsule synthesis. It remains to be determined whether the expression of these very different surface components occurs simply via random events of phase variation or in a coordinated manner in response to specific environmental cues. PMID- 16040976 TI - Toll-like receptor 4 mediates tolerance in macrophages stimulated with Toxoplasma gondii-derived heat shock protein 70. AB - Peritoneal macrophages (PMs) from toll-like receptor 4 (TLR4)-deficient and wild type (WT) mice were responsive to recombinant Toxoplasma gondii-derived heat shock protein 70 (rTgHSP70) and natural TgHSP70 (nTgHSP70) in NO release, but those from TLR2-, myeloid differentiation factor 88 (MyD88)-, and interleukin-1R associated kinase 4 (IRAK4)-deficient mice were not. Polymyxin B did not inhibit PM activation by TgHSP70 and nTgHSP70 from WT and TLR4-deficient mice, while it inhibited PM activation by lipopolysaccharide. Pretreatment of PMs from WT but not from TLR4-deficient mice with rTgHSP70 resulted in suppression of NO release on restimulation with rTgHSP70. Similarly, pretreatment of PMs from WT but not TLR4-deficient mice with nTgHSP70 resulted in suppression of NO release on restimulation with nTgHSP70. Polymyxin B did not inhibit rTgHSP70- and nTgHSP70 induced tolerance of PMs from TLR4-deficient mice. Furthermore, PMs from WT mice increased suppressor of cytokine-signaling-1 (SOCS-1) expression after restimulation with rTgHSP70, while those from TLR4-deficient mice did not. Phosphorylation of JNK and I-kappaBalpha occurred in rTgHSP70-induced tolerance of PMs from TLR4-deficient mice, but not in that from WT mice. These data indicated that TgHSP70 signaling mechanisms were mediated by TLR2, MyD88, and IRAK4, but not by TLR4. On the other hand, signaling of TgHSP70-induced tolerance was mediated by TLR4, and the expression of SOCS-1 suppressed the TLR2 signaling pathway. PMID- 16040977 TI - Analysis of cell type-specific responses mediated by the type IV secretion system of Helicobacter pylori. AB - Helicobacter pylori persistently infects the human stomach and can cause gastritis, gastric ulceration, and gastric cancer. The type IV secretion system (TFSS) of virulent H. pylori strains translocates the CagA protein, inducing the dephosphorylation of host cell proteins and leading to changes in the morphology or shape of AGS gastric epithelial cells. Furthermore, the TFSS is involved in the induction of proinflammatory cytokines. While the H. pylori genes required for TFSS function have been investigated systematically, little is known about possible host cell factors involved. We infected 19 different mammalian cell lines individually with H. pylori and analyzed CagA translocation, dephosphorylation of host cell proteins, chemokine secretion (interleukin-8 and macrophage inflammatory protein 2), and changes in cellular phenotypes. Our results demonstrate that not only bacterial but also host cell factors determine the cellular response to infection. The identification of such unknown host cell factors will add to our understanding of host-pathogen interactions and might help in the development of new therapeutic strategies. PMID- 16040978 TI - Illustration of pneumococcal polysaccharide capsule during adherence and invasion of epithelial cells. AB - The capsular polysaccharide of Streptococcus pneumoniae represents an important virulence factor and protects against phagocytosis. In this study the amount of capsular polysaccharide present on the bacterial surface during the infection process was illustrated by electron microscopic studies. After infection of A549 cells (type II pneumocytes) and HEp-2 epithelial cells a modified fixation method was used that allowed visualization of the state of capsule expression. This modified fixation procedure did not require the use of capsule-specific antibodies. Visualization of pneumococci in intimate contact and invading cells demonstrated that pneumococci were devoid of capsular polysaccharide. Pneumococci not in contact with the cells did not show alterations in capsular polysaccharide. After infection of the cells, invasive pneumococci of different strains and serotypes were recovered. Single colonies of these recovered pneumococci exhibited an up-to-10(5)-fold-enhanced capacity to adhere and an up to-10(4)-fold-enhanced capacity to invade epithelial cells. Electron microscopic studies using a lysine-ruthenium red (LRR) fixation procedure or cryo-field emission scanning electron microscopy revealed a reduction in capsular material, as determined in detail for a serotype 3 pneumococcal strain. The amount of polysaccharide in the serotype 3 capsule was also determined after intranasal infection of mice. This study illustrates for the first time the phenotypic variation of the polysaccharide capsule in the initial phase of pneumococcal infections. The modified LRR fixation allowed monitoring of the state of capsule expression of pathogens during the infectious process. PMID- 16040979 TI - Exposure to rumen protozoa leads to enhancement of pathogenicity of and invasion by multiple-antibiotic-resistant Salmonella enterica bearing SGI1. AB - Multiple-antibiotic-resistant Salmonella enterica serotype Typhimurium is a food borne pathogen that has been purported to be more virulent than antibiotic sensitive counterparts. The paradigm for this multiresistant/hyperpathogenic phenotype is Salmonella enterica serotype Typhimurium phage type DT104 (DT104). The basis for the multiresistance in DT104 is related to an integron structure designated SGI1, but factors underlying hyperpathogenicity have not been completely identified. Since protozoa have been implicated in the alteration of virulence in Legionella and Mycobacterium spp., we attempted to assess the possibility that protozoa may contribute to the putative hypervirulence of DT104. Our study reveals that DT104 can be more invasive, as determined by a tissue culture invasion assay, after surviving within protozoa originating from the bovine rumen. The enhancement of invasion was correlated with hypervirulence in a bovine infection model in which we observed a more rapid progression of disease and a greater recovery rate for the pathogen. Fewer DT104 cells were recovered from tissues of infected animals when protozoa were lysed by preinfection chemical defaunation of the bovine or ovine rumen. The protozoan-mediated hypervirulence phenotype was observed only in DT104 and other Salmonella strains, including serovars Agona and Infantis, possessing SGI1. PMID- 16040980 TI - Enhancing the protective efficacy of Mycobacterium bovis BCG vaccination against tuberculosis by boosting with the Mycobacterium tuberculosis major secretory protein. AB - Tuberculosis continues to ravage humanity, killing 2 million people yearly. Most cases occur in areas of the world to which the disease is endemic, where almost everyone is vaccinated early in life with Mycobacterium bovis BCG, the currently available vaccine against tuberculosis. Thus, while more-potent vaccines are needed to replace BCG, new vaccines are also needed to boost the immune protection of the 4 billion people already vaccinated with BCG. Until now, no booster vaccine has been shown capable of significantly enhancing the level of protective immunity induced by BCG in the stringent guinea pig model of pulmonary tuberculosis, the "gold standard" for testing tuberculosis vaccines. In this paper, we describe a booster vaccine for BCG comprising the purified recombinant Mycobacterium tuberculosis 30-kDa protein, the major secreted protein of this pathogen. In the guinea pig model of pulmonary tuberculosis, boosting BCG immunized animals once with the 30-kDa protein greatly increased cell-mediated and humoral immune responses to the protein in three consecutive experiments. Most importantly, boosting BCG-immunized animals once with the 30-kDa protein significantly enhanced protective immunity against aerosol challenge with highly virulent M. tuberculosis, as evidenced by a significantly reduced lung and spleen burden of M. tuberculosis compared with those for nonboosted BCG-immunized animals (mean additional reduction in CFU of 0.4 +/- 0.1 log in the lung [P = 0.03] and 0.6 +/- 0.1 log in the spleen [P = 0.002]). This study suggests that administering BCG-immunized people a booster vaccine comprising the 30-kDa protein may enhance their level of immunoprotection against tuberculosis. PMID- 16040981 TI - T-cell-stimulating protein A elicits immune responses during meningococcal carriage and human disease. AB - In recognition of the need for immunological memory-inducing components for future Neisseria meningitidis group B vaccines, we previously searched the proteome of N. meningitidis and identified T-cell-stimulating protein A (TspA). This study was designed to confirm the immunogencity of TspA and to examine the subset of T-helper cell responses to the protein in patients and nasopharyngeal carriers. The tspA gene was reconstructed, cloned, and expressed in Escherichia coli, and the recombinant TspA (rTspA) protein was affinity purified. T-cell proliferative responses to rTspA were detected in the peripheral blood mononuclear cells (PBMCs) of convalescent patients and carriers, confirming that TspA-specific T-cell responses were stimulated by invasive disease and nasopharyngeal colonization. Following stimulation of PBMCs with meningococcal lysate, increased frequencies of both Th1 and Th2 cells were observed, indicating that, as during carriage, invasive meningococcal disease induced an unbiased T helper subset response. A similar unbiased T-helper response was also detected against rTspA in the PBMCs of convalescent patients. The response of PBMCs from the carriers to TspA stimulation, however, was very weak, and the frequencies of cytokine-positive CD4 cells were not significantly greater than the frequencies in unstimulated control cultures. All of the patients and carriers responded with serum antimeningococcal immunoglobulin G (IgG) antibodies, while four of six samples from patients and 5 of 14 samples from carriers contained detectable anti rTspA IgG antibodies. Taken together, the results of this study confirmed the immunogenicity of TspA in humans during natural meningococcal infection, and therefore, TspA is worthy of further investigation as a possible T-cell stimulating component of future vaccines. PMID- 16040982 TI - Protection by natural human immunoglobulin M antibody to meningococcal serogroup B capsular polysaccharide in the infant rat protection assay is independent of complement-mediated bacterial lysis. AB - Neisseria meningitidis, an important cause of bacterial meningitis and septicemia worldwide, is associated with high mortality and serious sequelae. Natural immunity against meningococcal disease develops with age, but the specificity and functional activity of natural antibodies associated with protection are poorly understood. We addressed this question by using a selected subset of prevaccination sera (n = 26) with convergent or discrepant serum bactericidal activity (SBA) and infant rat protective activity (IRPA) against the serogroup B meningococcal strain 44/76-SL (B:15:P1.7,16) from Icelandic teenagers. The sera were analyzed by opsonophagocytic activity (OPA) assay, immunoblotting, immunoglobulin G (IgG) quantitation against live meningococcal cells by flow cytometry, and enzyme immunosorbent assay (EIA). High levels of SBA and OPA were reflected in distinct IgG binding to major outer membrane proteins and/or lipopolysaccharide in immunoblots. However, we could not detect any specific antibody patterns on blots that could explain IRPA. Only IgM antibody to group B capsular polysaccharide (B-PS), measured by EIA, correlated positively (r = 0.76, P < 0.001) with IRPA. Normal human sera (NHS; n = 20) from healthy Finnish children of different ages (7, 14, and 24 months and 10 years) supported this finding and showed an age-related increase in IRPA that coincided with the acquisition of B-PS specific IgM antibody. The protection was independent of complement-mediated bacterial lysis, as detected by the inability of NHS to augment SBA in the presence of human or infant rat complement and the equal protective activity of NHS in rat strains with fully functional or C6-deficient complement. PMID- 16040983 TI - Fungal antigens expressed during invasive aspergillosis. AB - Rabbits that had been infected intravenously with conidiospores of Aspergillus fumigatus were used as sources of antibody for screening a lambda phage cDNA expression library. The cDNA was derived from A. fumigatus mRNA that had been extracted from newly formed, germling hyphae. Thirty-six antigens were identified using antisera from six rabbits. Though many of these antigens were expected to be intracellular proteins because their genes did not encode a signal sequence, the antisera showed consistently a stronger immunoblot reaction with a cell fraction enriched for the fungal cell wall than with a fraction of predominantly intracellular components. Antibodies to eight antigens, including the glycosylhydrolase Asp f 16, were produced by more than one rabbit. In current vaccine studies, Asp f 16 is the only single antigen which has been reported to be capable of inducing protection against invasive aspergillosis in mice. Enolase and Aspergillus HSP90 were detected also; their homologues in Candida albicans have been tested as vaccines and have been reported to provide a partially protective response against invasive candidiasis in mice. The Aspergillus antigens reported here may have value both in diagnostic tests for different forms of aspergillosis and as vaccine candidates for protection against invasive disease. PMID- 16040984 TI - Conditions influencing the efficacy of vaccination with live organisms against Leishmania major infection. AB - Numerous experimental vaccines have been developed with the goal of generating long-term cell-mediated immunity to the obligate intracellular parasite Leishmania major, yet inoculation with live, wild-type L. major remains the only successful vaccine in humans. We examined the expression of immunity at the site of secondary, low-dose challenge in the ear dermis to determine the kinetics of parasite clearance and the early events associated with the protection conferred by vaccination with live L. major organisms in C57BL/6 mice. Particular attention was given to the route of vaccination. We observed that the rapidity, strength, and durability of the memory response following subcutaneous vaccination with live parasites in the footpad are even greater than previously appreciated. Antigen-specific gamma interferon (IFN-gamma)-producing T cells infiltrate the secondary site by 1.5 weeks, and viable parasites are cleared as early as 2.5 weeks following rechallenge, followed by a rapid drop in IFN-gamma(+) CD4(+) cell numbers in the site. In comparison, intradermal vaccination with live parasites in the ear generates immunity that is delayed in effector cell recruitment to the rechallenge site and in the clearance of parasites from the site. This compromised immunity was associated with a rapid recruitment of interleukin-10 (IL-10)-producing CD4(+) T cells to the rechallenge site. Treatment with anti-IL 10-receptor or anti-CD25 antibody enhanced early parasite clearance in ear vaccinated mice, indicating that chronic infection in the skin generates a population of regulatory cells capable of influencing the level of resistance to reinfection. A delicate balance of effector and regulatory T cells may be required to optimize the potency and durability of vaccines against Leishmaniasis and other intracellular pathogens. PMID- 16040985 TI - Growth characteristics of and virulence factor production by group A Streptococcus during cultivation in human saliva. AB - Group A Streptococcus (GAS) commonly infects the human oropharynx, but the initial molecular events governing this process are poorly understood. Saliva is a major component of the innate and acquired immune defense in this anatomic site. Although landmark studies were done more than 60 years ago, investigation of GAS-saliva interaction has not been addressed extensively in recent years. Serotype M1 GAS strain MGAS5005 cultured in human saliva grew to approximately 10(7) CFU/ml and, remarkably, maintained this density for up to 28 days. Strains of several other M-protein serotypes had similar initial growth patterns but did not maintain as high a CFU count during prolonged culture. As revealed by analysis of the growth of isogenic mutant strains, the ability of GAS to maintain high numbers of CFU/ml during the prolonged stationary phase in saliva was dependent on production of streptococcal inhibitor of complement (Sic) and streptococcal pyrogenic exotoxin B (SpeB). During cultivation in human saliva, GAS had growth-phase-dependent production of multiple proven and putative extracellular virulence factors, including Sic, SpeB, streptococcal pyrogenic exotoxin A, Mac protein, and streptococcal phospholipase A(2). Our results clearly show that GAS responds in a complex fashion to growth in human saliva, suggesting that the molecular processes that enhance colonization and survival in the upper respiratory tract of humans are well under way before the organism reaches the epithelial cell surface. PMID- 16040986 TI - Characterization of the Helicobacter pylori cysteine-rich protein A as a T-helper cell type 1 polarizing agent. AB - Predominant T-helper 1 (Th1) responses with increased gamma interferon (IFN gamma) levels have been proposed to play an important role in Helicobacter pylori induced gastritis and peptic ulceration. However, bacterial factors contributing to the initiation of Th1 polarization of H. pylori-specific immune responses have not been characterized in detail thus far. We report here on the identification of Helicobacter cysteine-rich protein A (HcpA) as a novel proinflammatory and Th1 promoting protein. The capacity of HcpA to induce immune activation was studied in splenocyte cultures of naive H. pylori-negative mice. HcpA stimulated the release of high concentrations of the proinflammatory and Th1-promoting cytokines interleukin-6 (IL-6) and IFN-gamma, in addition to significant levels of IL-12, tumor necrosis factor alpha, and IL-10. The observed cytokine profile was comparable to that induced by lipopolysaccharide but differed in the kinetics and maximum levels of cytokine production. In addition, HcpA-induced cytokine release resembled that observed upon incubation with H. pylori except for IL-10, which was only moderately released upon HcpA stimulation. Both HcpA- and H. pylori mediated IFN-gamma production was drastically reduced by a neutralizing antibody against IL-12 but not by an anti-IL-2 antibody. Thus, HcpA seems to represent a novel bacterial virulence factor triggering the release of a concerted set of cytokines to instruct the adaptive immune system for the initiation of proinflammatory and Th1-biased immunity. PMID- 16040987 TI - Influence of Na(+), dicarboxylic amino acids, and pH in modulating the low calcium response of Yersinia pestis. AB - The virulence of yersiniae is promoted in part by shared approximately 70-kb plasmids (pCD in Yersinia pestis and pYV in enteropathogenic Yersinia pseudotuberculosis and Yersinia enterocolitica) that mediate a low-calcium response. This phenotype is characterized at 37 degrees C by either bacteriostasis in Ca(2+)-deficient medium with expression of pCD/pYV-encoded virulence effectors (Yops and LcrV) or vegetative growth and repression of Yops and LcrV with > or =2.5 mM Ca(2+) (Lcr(+)). Regulation of Yops and LcrV is well defined but little is known about bacteriostasis other than that Na(+) plus l glutamate promotes prompt restriction of Y. pestis. As shown here, l-aspartate substituted for l-glutamate in this context but only Na(+) exacerbated the nutritional requirement for Ca(2+). Bacteriostasis of Y. pestis (but not enteropathogenic yersiniae) was abrupt in Ca(2+)-deficient medium at neutral to slightly alkaline pH (7.0 to 8.0), although increasing the pH to 8.5 or 9.0, especially with added Na(+) (but not l-glutamate), facilitated full-scale growth. Added l-glutamate (but not Na(+)) favored Ca(2+)-independent growth at acidic pH (5.0 to 6.5). Yops and LcrV were produced in Ca(2+)-deficient media at pH 6.5 to 9.0 regardless of the presence of added Na(+) or l-glutamate, although their expression at alkaline pH was minimal. Resting Ca(2+)-starved Lcr(+) cells of Y. pestis supplied with l-glutamate first excreted and then destroyed l-aspartate. These findings indicate that expression of Yops and LcrV is necessary but not sufficient for bacteriostasis of Ca(2+)-starved yersiniae and suggest that abrupt restriction of Y. pestis requires Na(+) and the known absence of aspartate ammonia-lyase in this species. PMID- 16040988 TI - Identification and characterization of the locus for diffuse adherence, which encodes a novel afimbrial adhesin found in atypical enteropathogenic Escherichia coli. AB - The O26 serogroup of enteropathogenic Escherichia coli (EPEC) is one of the serogroups most frequently implicated in infant diarrhea and is also common among enterohemorrhagic E. coli (EHEC) strains. The most common O26 strains belong to EPEC/EHEC serotype O26:H11 and are generally Shiga toxin (Stx) positive. Stx negative E. coli strains that are negative for the EPEC EAF plasmid and bundle forming pilus (Bfp) are classified as atypical EPEC. Here, we report a novel adhesin present in an stx-negative bfpA-negative atypical EPEC O26:H11 strain isolated from an infant with diarrhea. A cloned 15-kb genomic region from this strain, designated the locus for diffuse adherence (lda), confers diffuse adherence on HEp-2 cells when expressed in E. coli K-12. Sequence analysis of lda revealed a G+C content of 46.8% and 15 open reading frames sharing homology with the E. coli K88 fae and CS31A clp fimbrial operons. The lda region is part of a putative 26-kb genomic island inserted into the proP gene of the E. coli chromosome. Hybridization studies have demonstrated the prevalence of the minor structural subunit gene, ldaH, across E. coli serogroups O5, O26, O111, and O145. A second plasmid-encoded factor that contributed to the Hep-2 adherence of this strain was also identified but was not characterized. Null mutations that abolish adherence to HEp-2 cells can be restored by plasmid complementation. Antiserum raised against the major structural subunit, LdaG, recognizes a 25-kDa protein from crude heat-extracted protein preparations and inhibits the adherence of the E. coli DH5alpha lda(+) clone to HEp-2 cells. Electron microscopy revealed a nonfimbrial structure surrounding the bacterial cell. PMID- 16040989 TI - Pathoadaptive mutation that mediates adherence of shiga toxin-producing Escherichia coli O111. AB - The adherence of pathogenic Escherichia coli strains to intestinal epithelium is essential for initiation of infection. The cad operon encodes the lysine decarboxylase (LDC) system responsible for metabolizing lysine, and this operon has been proposed as an antivirulence mechanism in enteroinvasive E. coli and Shigella flexneri and as a factor mediating E. coli O157:H7 adherence. We sought to determine whether the LDC activity was present in a phylogenetically characterized collection of diarrheagenic E. coli (DEC) strains and to establish whether its expression was associated with their adherence to tissue culture cells. LDC activity was found in most of the pathogenic E. coli strains tested and was absent from Shiga toxin-producing E. coli (STEC) O111 strains (DEC pathotype 8). Analysis of the cad region in these O111 strains indicates that the operon has been rearranged and some of the genes are either missing or disrupted. A similar rearrangement was found in an E. coli O111:H8 strain recently isolated from an outbreak in Texas. Complementation of the LDC-negative strains with the cad operon in trans restored the LDC activity and resulted in a reduction in adherence to tissue culture cells. Initial analysis of the protein profiles on the surface of the O111 strains indicates that the LDC activity has an effect on the expression of the adhesin intimin. Cadaverine had a slight effect on LDC negative strain adhesion but none on intimin expression. Our data suggest that this pathoadaptive mutation is an important mechanism to control functions potentially implicated in the pathogenesis of these organisms. PMID- 16040990 TI - Species-specific inhibition of cerebral malaria in mice coinfected with Plasmodium spp. AB - Recent epidemiological observations suggest that clinical evolution of Plasmodium falciparum infections might be influenced by the concurrent presence of another Plasmodium species, and such mixed-species infections are now known to occur frequently in residents of most areas of endemicity. We used mice infected with P. berghei ANKA (PbA), a model for cerebral malaria (CM), to investigate the influence of experimental mixed-species infections on the expression of this pathology. Remarkably, the development of CM was completely inhibited by the simultaneous presence of P. yoelii yoelii but not that of P. vinckei or another line of P. berghei. In the protected coinfected mice, the accumulation of CD8(+) T cells in the brain vasculature, a pivotal step in CM pathogenesis, was found to be abolished. Protection from CM was further found to be associated with species specific suppression of PbA multiplication. These observations establish the concept of mixed Plasmodium species infections as potential modulators of pathology and open novel avenues to investigate mechanisms implicated in the pathogenesis of malaria. PMID- 16040991 TI - Independent protective effects for tumor necrosis factor and lymphotoxin alpha in the host response to Listeria monocytogenes infection. AB - Although the essential role of tumor necrosis factor (TNF) in resistance to Listeria monocytogenes infection is well established, the roles of the related cytokines lymphotoxin alpha (LTalpha) and lymphotoxin beta (LTbeta) are unknown. Using C57BL/6 mice in which the genes for these cytokines were disrupted, we examined the contributions of TNF, LTalpha, and LTbeta in the host response to Listeria. To overcome the lack of peripheral lymph nodes in LTalpha(-/-) and LTbeta(-/-) mice, bone marrow chimeras were constructed. TNF(-/-) and LTalpha(-/ ) chimeras that lacked both secreted LTalpha(3) and membrane-bound LTalpha(1)beta(2) and LTalpha(2)beta(1) were highly susceptible and succumbed 4.5 and 6 days, respectively, after a low-dose infection (200 CFU). LTbeta(-/-) chimeras, which lacked only membrane-bound LT, controlled the infection in a manner comparable to wild-type (WT) chimeras. The Listeria-specific proliferative and gamma interferon T-cell responses were equivalent in all five groups of infected mice (LTalpha(-/-) and LTbeta(-/-) chimeras, WT chimeras, and TNF(-/-) and WT mice). TNF(-/-) mice and LTalpha(-/-) chimeras, however, failed to generate the discrete foci of lymphocytes and macrophages that are essential for bacterial elimination. Rather, aberrant necrotic lesions comprised predominantly of neutrophils with relatively few lymphocytes and macrophages were observed in the livers and spleens of TNF(-/-) and LTalpha(-/-) chimeras. Therefore, in addition to TNF, soluble LTalpha(3) plays a separate essential role in control of listerial infection through control of leukocyte accumulation and organization in infected organs. PMID- 16040992 TI - The multifunctional Staphylococcus aureus autolysin aaa mediates adherence to immobilized fibrinogen and fibronectin. AB - Staphylococci can cause a wide spectrum of infections, including endocarditis, osteomyelitis, and sepsis, which is reflected by the numerous virulence factors they produce, among them a recently identified new class of adhesins, namely, the multifunctional autolysins/adhesins. Here we report the identification and molecular characterization of Aaa, a novel autolysin/adhesin from Staphylococcus aureus. The gene encoding Aaa was cloned from the clinical isolate Staphylococcus aureus 4074. DNA sequence analysis revealed that aaa encodes a deduced protein of 334 amino acids with a predicted molecular mass of 35.8 kDa. Aaa contains three N terminal repetitive sequences that comprise features of a peptidoglycan-binding domain, the LysM domain. The expression of aaa by Escherichia coli and its subsequent characterization revealed that Aaa possesses bacteriolytic activity as well as adhesive properties, such as binding to extracellular matrix proteins. Real-time biomolecular interaction analysis demonstrated that the interaction of Aaa with fibrinogen, fibronectin, and vitronectin is dose dependent and saturable and occurs with a high affinity. Furthermore, we demonstrate that Aaa binds to the Aalpha and Bbeta chains of fragment D of fibrinogen. Immunofluorescence microscopy revealed that Aaa is located at the cell surface. Finally, an aaa knockout mutant showed reduced adherence to surface-adsorbed fibrinogen and fibronectin, strongly suggesting a role for Aaa in the colonization of host factor-coated polymer surfaces and/or host tissue. PMID- 16040993 TI - Effect of skin barrier disruption on immune responses to topically applied cross reacting material, CRM(197), of diphtheria toxin. AB - The high accessibility of the skin and the presence of immunocompetent cells in the epidermis makes this surface an attractive route for needle-free administration of vaccines. However, the lining of the skin by the stratum corneum is a major obstacle to vaccine delivery. In this study we examined the effect of skin barrier disruption on the immune responses to the cross-reacting material CRM(197), a nontoxic mutant of diphtheria toxin (DTx) that is considered as a vaccine candidate. Application of CRM(197), together with cholera toxin (CT), onto the tape-stripped skin of mice elicited antibody responses that had anti-DTx neutralizing activity. Vaccine delivery onto mildly ablated skin or intact skin did not elicit any detectable anti-CRM(197) antibodies. Mice immunized with CRM(197) alone onto the tape-stripped skin mounted a vigorous antigen-specific proliferative response. In contrast, the induction of cellular immunity after CRM(197) deposition onto mildly ablated or intact skin was adjuvant dependent. Furthermore, epidermal cells were activated and underwent apoptosis that was more pronounced when the stratum corneum was removed by tape stripping. Overall, these findings highlight the potential for transcutaneous delivery of CRM(197) and establish a correlation between the degree of barrier disruption and levels of antigen-specific immune responses. Moreover, these results provide the first evidence that the development of a transcutaneous immunization strategy for diphtheria, based on simple and practical methods to disrupt the skin barrier, is feasible. PMID- 16040994 TI - Bacterial ghosts as an oral vaccine: a single dose of Escherichia coli O157:H7 bacterial ghosts protects mice against lethal challenge. AB - Enterohemorrhagic Escherichia coli (EHEC) is a bacterial pathogen that is associated with several life-threatening diseases for humans. The combination of protein E-mediated cell lysis to produce EHEC ghosts and staphylococcal nuclease A to degrade DNA was used for the development of an oral EHEC vaccine. The lack of genetic material in the oral EHEC bacterial-ghost vaccine abolished any hazard of horizontal gene transfer of resistance genes or pathogenic islands to resident gut flora. Intragastric immunization of mice with EHEC ghosts without the addition of any adjuvant induced cellular and humoral immunity. Immunized mice challenged at day 55 showed 86% protection against lethal challenge with a heterologous EHEC strain after single-dose oral immunization and 93.3% protection after one booster at day 28, whereas the controls showed 26.7% and 30% survival, respectively. These results indicate that it is possible to develop an efficacious single-dose oral EHEC bacterial-ghost vaccine. PMID- 16040995 TI - Slow genetic divergence of Helicobacter pylori strains during long-term colonization. AB - The genetic variability of Helicobacter pylori is known to be high compared to that of many other bacterial species. H. pylori is adapted to the human stomach, where it persists for decades, and adaptation to each host results in every individual harboring a distinctive bacterial population. Although clonal variants may exist within such a population, all isolates are generally genetically related and thus derived from a common ancestor. We sought to determine the rate of genetic change of H. pylori over 9 years in two asymptomatic adult patients. Arbitrary primed PCR confirmed the relatedness of individual subclones within a patient. Furthermore, sequencing of 10 loci ( approximately 6,000 bp) in three subclones per time and patient revealed only two base pair changes among the subclones from patient I. All sequences were identical among the patient II subclones. However, PCR amplification of the highly divergent gene amiA revealed great variation in the size of the gene between the subclones within each patient. Thus, both patients harbored a single strain with clonal variants at both times. We also studied genetic changes in culture- and mouse-passaged strains, and under both conditions no genetic divergence was found. These results suggest that previous estimates of the rate of genetic change in H. pylori within an individual might be overestimates. PMID- 16040996 TI - Distinct defensin profiles in Neisseria gonorrhoeae and Chlamydia trachomatis urethritis reveal novel epithelial cell-neutrophil interactions. AB - Defensins are key participants in mucosal innate defense. The varied antimicrobial activity and differential distribution of defensins at mucosal sites indicate that peptide repertoires are tailored to site-specific innate defense requirements. Nonetheless, few studies have investigated changes in peptide profiles and function after in vivo pathogen challenge. Here, we determined defensin profiles in urethral secretions of healthy men and men with Chlamydia trachomatis- and Neisseria gonorrhoeae-mediated urethritis by immunoblotting for the epithelial defensins HBD1, HBD2, and HD5 and the neutrophil defensins HNP1 to -3 (HNP1-3). HBD1 was not detectable in secretions, and HBD2 was only induced in a small proportion of the urethritis patients; however, HD5 and HNP1-3 were increased in C. trachomatis infection and significantly elevated in N. gonorrhoeae infection. When HNP1-3 levels were low, HD5 appeared mostly as the propeptide; however, when HNP1-3 levels were >10 microg/ml, HD5 was proteolytically processed, suggesting neutrophil proteases might contribute to HD5 processing. HD5 and HNP1-3 were bactericidal against C. trachomatis and N. gonorrhoeae, but HD5 activity was dependent upon N-terminal processing of the peptide. In vitro proteolysis of proHD5 by neutrophil proteases and analysis of urethral secretions by surface-enhanced laser desorption ionization substantiated that neutrophils contribute the key convertases for proHD5 in the urethra during these infections. This contrasts with the small intestine, where Paneth cells secrete both proHD5 and its processing enzyme, trypsin. In conclusion, we describe a unique defensin expression repertoire in response to inflammatory sexually transmitted infections and a novel host defense mechanism wherein epithelial cells collaborate with neutrophils to establish an antimicrobial barrier during infection. PMID- 16040997 TI - Global gene expression and the role of sigma factors in Neisseria gonorrhoeae in interactions with epithelial cells. AB - Like many bacterial pathogens, Neisseria gonorrhoeae must adapt to environmental changes in order to successfully colonize and proliferate in a new host. Modulation of gene expression in response to environmental signals is an efficient mechanism used by bacteria to achieve this goal. Using DNA microarrays and a tissue culture model for gonococcal infection, we examined global changes in gene expression in N. gonorrhoeae in response to adherence to host cells. Among those genes induced upon adherence to human epithelial cells in culture was rpoH, which encodes a homolog of the heat shock sigma factor, sigma(32) (RpoH), as well as genes of the RpoH regulon, groEL and groES. Attempts to construct an rpoH null mutant in N. gonorrhoeae were unsuccessful, suggesting that RpoH is essential for viability of N. gonorrhoeae. The extracytoplasmic sigma factor, RpoE (sigma(E)), while known to regulate rpoH in other bacteria, was found not to be necessary for the up-regulation of rpoH in gonococci upon adherence to host cells. To examine the role of RpoH in host cell interactions, an N. gonorrhoeae strain conditionally expressing rpoH was constructed. The results of our experiments showed that while induction of rpoH expression is not necessary for adherence of gonococci to epithelial cells, it is important for the subsequent invasion step, as gonococci depleted for rpoH invade cells two- to threefold less efficiently than a wild-type strain. Taken together, these results indicate that sigma(32), but not sigma(E), is important for the response of gonococci in the initial steps of an infection. PMID- 16040998 TI - Involvement of ganglioside GM3 in G(2)/M cell cycle arrest of human monocytic cells induced by Actinobacillus actinomycetemcomitans cytolethal distending toxin. AB - Actinobacillus actinomycetemcomitans produces a toxin called cytolethal distending toxin (CDT), which causes host cell DNA damage leading to the induction of DNA damage checkpoint pathways. CDT consists of three subunits, CdtA, CdtB, and CdtC. CdtB is the active subunit of CDT and exerts its effect as a nuclease that damages nuclear DNA, triggering cell cycle arrest. In the present study, we confirmed that the only combination of toxin proteins causing cell cycle arrest was that of all three recombinant CDT (rCDT) protein subunits. Furthermore, in order for rCDT to demonstrate toxicity, it was necessary for CdtA and CdtC to access the cell before CdtB. The coexistence of CdtA and CdtC was necessary for these subunits to bind to the cell. Cells treated with the glucosylceramide synthesis inhibitor 1-phenyl-2-palmitoylamino-3-morpholino-1 propanol showed resistance to the cytotoxicity induced by rCDT. Furthermore, LY-B cells, which are deficient in the biosynthesis of sphingolipid, also showed resistance to the cytotoxicity induced by rCDT. To evaluate the binding of each subunit for glucosylceramides, we performed thin-layer chromatography immunostaining. The results indicated that each subunit reacted with the glycosphingolipids GM1, GM2, GM3, Gb3, and Gb4. The rCDT mixture incubated with liposomes containing GM3 displayed partially reduced toxicity. These results indicate that GM3 can act as a CDT receptor. PMID- 16040999 TI - Surfaceome of Leptospira spp. AB - The identification of the subset of outer membrane proteins exposed on the surface of a bacterial cell (the surfaceome) is critical to understanding the interactions of bacteria with their environments and greatly narrows the search for protective antigens of extracellular pathogens. The surfaceome of Leptospira was investigated by biotin labeling of viable leptospires, affinity capture of the biotinylated proteins, two-dimensional gel electrophoresis, and mass spectrometry (MS). The leptospiral surfaceome was found to be predominantly made up of a small number of already characterized proteins, being in order of relative abundance on the cell surface: LipL32 > LipL21 > LipL41. Of these proteins, only LipL32 had not been previously identified as surface exposed. LipL32 surface exposure was subsequently verified by three independent approaches: surface immunofluorescence, whole-cell enzyme-linked immunosorbent assay (ELISA), and immunoelectron microscopy. Three other proteins, Q8F8Q0 (a putative transmembrane outer membrane protein) and two proteins of 20 kDa and 55 kDa that could not be identified by MS, one of which demonstrated a high degree of labeling potentially representing an additional, as-yet-uncharacterized, surface-exposed protein. Minor labeling of p31(LipL45), GroEL, and FlaB1 was also observed. Expression of the surfaceome constituents remained unchanged under a range of conditions investigated, including temperature and the presence of serum or urine. Immunization of mice with affinity-captured surface components stimulated the production of antibodies that bound surface proteins from heterologous leptospiral strains. The surfaceomics approach is particularly amenable to protein expression profiling using small amounts of sample (<10(7) cells) offering the potential to analyze bacterial surface expression during infection. PMID- 16041000 TI - Expression of Arg-Gingipain RgpB is required for correct glycosylation and stability of monomeric Arg-gingipain RgpA from Porphyromonas gingivalis W50. AB - Arg-gingipains are extracellular cysteine proteases produced by the gram-negative periodontal pathogen Porphyromonas gingivalis and are encoded by rgpA and rgpB. Three Arg-gingipains, heterodimeric high-molecular-mass Arg-gingipain HRgpA comprising the alpha-catalytic chain and the beta-adhesin chain, the monomeric soluble Arg-gingipain comprising only the alpha-catalytic chain (RgpA(cat)), and the monomeric membrane-type heavily glycosylated Arg-gingipain comprising the alpha-catalytic chain (mt-RgPA(cat)), are derived from rgpA. The monomeric enzymes contain between 14 and 30% carbohydrate by weight. rgpB encodes two monomeric enzymes, RgpB and mt-RgpB. Earlier work indicated that rgpB is involved in the glycosylation process, since inactivation of rgpB results in the loss of not only RgpB and mt-RgpB but also mt-RgpA(cat). This work aims to confirm the role of RgpB in the posttranslational modification of RgpA(cat) and the effect of aberrant glycosylation on the properties of this enzyme. Two-dimensional gel electrophoresis of cellular proteins from W50 and an inactivated rgpB strain (D7) showed few differences, suggesting that loss of RgpB has a specific effect on RgpA maturation. Inactivation of genes immediately upstream and downstream of rgpB had no effect on rgpA-derived enzymes, suggesting that the phenotype of the rgpB mutant is not due to a polar effect on transcription at this locus. Matrix assisted laser desorption ionization-time of flight analysis of purified RgpA(cat) from W50 and D7 strains gave identical peptide mass fingerprints, suggesting that they have identical polypeptide chains. However, RgpA(cat) from D7 strain had a higher isoelectric point and a dramatic decrease in thermostability and did not cross-react with a monoclonal antibody which recognizes a glycan epitope on the parent strain enzyme. Although it had the same total sugar content as the parent strain enzyme, there were significant differences in the monosaccharide composition and linking sugars. These data suggest that RgpB is required for the normal posttranslational glycosylation of Arg-gingipains derived from rgpA and that this process is required for enzyme stabilization. PMID- 16041001 TI - Clostridium perfringens epsilon-toxin increases permeability of single perfused microvessels of rat mesentery. AB - Epsilon-toxin, the primary virulence factor of Clostridium perfringens type D, causes mortality in livestock, particularly sheep and goats, in which it induces an often-fatal enterotoxemia. It is believed to compromise the intestinal barrier and then enter the gut vasculature, from which it is carried systemically, causing widespread vascular endothelial damage and edema. Here we used single perfused venular microvessels in rat mesentery, which enabled direct observation of permeability properties of the in situ vascular wall during exposure to toxin. We determined the hydraulic conductivity (L(p)) of microvessels as a measure of the response to epsilon-toxin. We found that microvessels were highly sensitive to toxin. At 10 microg ml(-1) the L(p) increased irreversibly to more than 15 times the control value by 10 min. At 0.3 microg ml(-1) no increase in L(p) was observed for up to 90 min. The toxin-induced increase in L(p) was consistent with changes in ultrastructure of microvessels exposed to the toxin. Those microvessels exhibited gaps either between or through endothelial cells where perfusate had direct access to the basement membrane. Many endothelial cells appeared necrotic, highly attenuated, and with dense cytoplasm. We showed that epsilon-toxin, in a time- and dose-dependent manner, rapidly and irreversibly compromised the barrier function of venular microvessel endothelium. The results conformed to the hypothesis that epsilon-toxin interacts with vascular endothelial cells and increases the vessel wall permeability by direct damage of the endothelium. PMID- 16041002 TI - Fsr-independent production of protease(s) may explain the lack of attenuation of an Enterococcus faecalis fsr mutant versus a gelE-sprE mutant in induction of endocarditis. AB - An Enterococcus faecalis gelE insertion disruption mutant (TX5128), which produces neither gelatinase (GelE) nor the cotranscribed (in the wild type) serine protease (SprE), was found to be attenuated in a rat endocarditis model with a significant decrease in the endocarditis induction rate versus wild-type E. faecalis OG1RF (GelE(+), SprE(+)). TX5266, which has a nonpolar deletion in fsrB and, like TX5128, is phenotypically GelE(-) under usual conditions, was also studied; fsrB is a homologue of agrB of staphylococci and participates in regulation of gelE-sprE expression. Unexpectedly, TX5266 approximated wild-type OG1RF in the endocarditis model and was significantly less attenuated than TX5128. This is in contrast to other models which have found fsr mutants to be as or more attenuated than TX5128. Further study found that the fsrB mutant produced very low levels of gelatinase activity after prolonged incubation in vitro versus no gelatinase activity with TX5128 and did not show the extensive chaining characteristic of TX5128. Reverse transcription-PCR confirmed that gelE was expressed in TX5266 at a very low level versus wild-type OG1RF and was not expressed at all in TX5128. Possible explanations for the increased induction of endocarditis by TX5266 versus TX5128 include the production of low levels of protease(s) or some other effect(s) of the inactivation of the E. faecalis fsr regulator. The equivalent ability of OG1RF and its fsr mutant to initiate endocarditis may explain why we did not find naturally occurring fsr mutants, which account for ca. 35% of E. faecalis isolates, unrepresented in endocarditis versus fecal isolates. PMID- 16041003 TI - Comparison of Helicobacter pylori virulence gene expression in vitro and in the Rhesus macaque. AB - We used a quantitative real-time reverse transcriptase PCR assay to measure the transcript abundance of 46 known and putative Helicobacter pylori virulence genes, including 24 genes on the Cag pathogenicity island. The expression profile of H. pylori cells grown in vitro was also compared to expression in vivo after experimental infection of rhesus macaques. Transcript abundance in vitro (mid-log phase) ranged from about 0.004 (feoB and hpaA) to 20 (ureAB, napA, and cag25) copies/cell. Expression of most genes was repressed during the transition from logarithmic- to stationary-phase growth, but several well-characterized H. pylori virulence genes (katA, napA, vacA, and cagA) were induced. Comparison of results in the rhesus macaque with similar data from humans showed a strong correlation (r = 0.89). The relative in vivo expression in the rhesus monkey was highly correlated with in vitro expression during mid-log (r = 0.89)- and stationary (r = 0.88)-phase growth. Transcript abundance was on average three- to fourfold reduced in vivo compared to in vitro during mid-log phase. However, when compared to stationary phase, increased expression in vivo was observed for 6 of 7 genes on a contiguous portion of the pathogenicity island, several of which are thought to encode the H. pylori type IV structural pilus and its accessory proteins. These results suggest the possibility that some genes encoding the H. pylori type IV structural pilus and accessory proteins may form an operon that is induced during growth in vivo. PMID- 16041004 TI - Divergence of protection induced by bacterial products and sepsis-induced immune suppression. AB - Susceptibility to bacterial infections after a primary immune stimulation differs drastically depending on the presensitization of the innate immune system. To determine the conditions that either induce protection or enhanced susceptibility to infection with Salmonella enterica serovar Typhimurium, we pretreated mice either with tumor necrosis factor (TNF), whole killed bacteria, or sublethal cecal ligation and puncture (CLP) as a mouse model for septic peritonitis. Impaired production of the cytokines TNF, interleukin-6 (IL-6), and IL-10 was induced by these pretreatment schedules, with TNF-signaling not being essential for this effect. Injection of TNF or killed bacteria enhanced survival of mice infected subsequently with serovar Typhimurium. In contrast, sepsis such as that induced by CLP only protected from shock induced by d-galactosamine and lipopolysaccharide or by a high dose of bacteria but sensitized to a secondary bacterial infection. Such sepsis-induced enhanced susceptibility to infection was critically dependent on TNF function. PMID- 16041005 TI - NK cells help to induce CD8(+)-T-cell immunity against Toxoplasma gondii in the absence of CD4(+) T cells. AB - CD8(+) T-cell immunity plays an important role in protection against intracellular infections. Earlier studies have shown that CD4(+) T-cell help was needed for launching in vivo CD8(+) T-cell activity against these pathogens and tumors. However, recently CD4(+) T-cell-independent CD8 responses during several microbial infections including those with Toxoplasma gondii have been described, although the mechanism is not understood. We now demonstrate that, in the absence of CD4(+) T cells, T. gondii-infected mice exhibit an extended NK cell response, which is mediated by continued interleukin-12 (IL-12) secretion. This prolonged NK cell response is critical for priming parasite-specific CD8(+) T-cell immunity. Depletion of NK cells inhibited the generation of CD8(+) T-cell immunity in CD4(-/-) mice. Similarly neutralization of IL-12 reduces NK cell numbers in infected animals and leads to the down-regulation of CD8(+) T-cell immunity against T. gondii. Adoptive transfer of NK cells into the IL-12-depleted animals restored their CD8(+) T-cell immune response, and animals exhibited reduced mortality. NK cell gamma interferon was essential for cytotoxic T lymphocyte priming. Our studies for the first time demonstrate that, in the absence of CD4(+) T cells, NK cells can play an important role in induction of primary CD8(+) T-cell immunity against an intracellular infection. These observations have therapeutic implications for immunocompromised individuals, including those with human immunodeficiency virus infection. PMID- 16041006 TI - Cryptococcus neoformans {alpha} strains preferentially disseminate to the central nervous system during coinfection. AB - Cryptococcus neoformans is a fungal pathogen that has evolved over the past 40 million years into three distinct varieties or sibling species (gattii, grubii, and neoformans). Each variety manifests differences in epidemiology and disease, and var. grubii strains are responsible for the vast majority of human disease. In previous studies, alpha strains were more virulent than congenic a strains in var. neoformans, whereas var. grubii congenic a and alpha strains exhibited equivalent levels of virulence. Here the role of mating type in the virulence of var. grubii was further characterized in a panel of model systems. Congenic var. grubii a and alpha strains had equivalent survival rates when cultured with amoebae, nematodes, and macrophages. No difference in virulence was observed between a and alpha congenic strains in multiple inbred-mouse genetic backgrounds, and there was no difference in accumulations in the central nervous system (CNS) late in infection. In contrast, during coinfections, a and alpha strains are equivalent in peripheral tissues but alpha cells have an enhanced predilection to penetrate the CNS. These studies reveal the first virulence difference between congenic a and alpha strains in the most common pathogenic variety and suggest an explanation for the prevalence of alpha strains in clinical isolates. PMID- 16041007 TI - Type 1 immunity provides both optimal mucosal and systemic protection against a mucosally invasive, intracellular pathogen. AB - It has been hypothesized that optimal vaccine immunity against mucosally invasive, intracellular pathogens may require the induction of different types of immune responses in mucosal and systemic lymphoid tissues. Mucosal type 2/3 responses (producing interleukin-4 [IL-4], IL-6 and/or transforming growth factor beta) could be necessary for optimal induction of protective secretory immunoglobulin A responses. On the other hand, systemic type 1 responses (including gamma interferon [IFN-gamma], tumor necrosis factor alpha, and optimal cytotoxic T-cell responses) are likely to be critical for protection against the disseminated intracellular replication that occurs after mucosal invasion. Despite these predictions, we recently found that vaccines inducing highly polarized type 1 immunity in both mucosal and systemic tissues provided optimal mucosal and systemic protection against the protozoan pathogen Trypanosoma cruzi. To further address this important question in a second model system, we now have studied the capacity of knockout mice to develop protective immune memory. T. cruzi infection followed by nifurtimox treatment rescue was used to immunize CD4, CD8, beta2-microglobulin, inducible nitric oxide synthase (iNOS), IL-12, IFN gamma, and IL-4 knockout mice. Despite the previously demonstrated importance of CD4(+) T cells, CD8(+) T cells, and nitric oxide for T. cruzi immunity, CD4, CD8, and iNOS knockout mice developed mucosal and systemic protective immunity. However, IL-12, IFN-gamma, and beta2-microglobulin-deficient mice failed to develop mucosal or systemic protection. In contrast, IL-4 knockout mice developed maximal levels of both mucosal and systemic immune protection. These results strongly confirm our earlier conclusion from studies with polarizing vaccination protocols that type 1 immunity provides optimal mucosal and systemic protection against a mucosally invasive, intracellular pathogen. PMID- 16041008 TI - Phagocyte-derived reactive oxygen species do not influence the progression of murine blood-stage malaria infections. AB - Phagocyte-derived reactive oxygen species have been implicated in the clearance of malaria infections. We investigated the progression of five different strains of murine malaria in gp91(phox-/-) mice, which lack a functional NADPH oxidase and thus the ability to produce phagocyte-derived reactive oxygen species. We found that the absence of functional NADPH oxidase in the gene knockout mice had no effect on the parasitemia or total parasite burden in mice infected with either resolving (Plasmodium yoelii and Plasmodium chabaudi K562) or fatal (Plasmodium berghei ANKA, Plasmodium berghei K173 and Plasmodium vinckei vinckei) strains of malaria. This lack of effect was apparent in both primary and secondary infections with P. yoelii and P. chabaudi. There was also no difference in the presentation of clinical or pathological signs between the gp91(phox-/-) or wild-type strains of mice infected with malaria. Progression of P. berghei ANKA and P. berghei K173 infections was unchanged in glutathione peroxidase-1 gene knockout mice compared to their wild-type counterparts. The rates of parasitemia progression in gp91(phox-/-) mice and wild-type mice were not significantly different when they were treated with l-N(G)-methylarginine, an inhibitor of nitric oxide synthase. These results suggest that phagocyte-derived reactive oxygen species are not crucial for the clearance of malaria parasites, at least in murine models. PMID- 16041009 TI - Prevention and treatment of cutaneous leishmaniasis in primates by using synthetic type D/A oligodeoxynucleotides expressing CpG motifs. AB - Oligodeoxynucleotides (ODN) containing CpG motifs mimic microbial DNA and are recognized by toll-like receptor 9 on immune cells. The resulting response limits the early spread of infectious organisms and promotes the development of adaptive immunity. In this regard, CpG ODN show promise as immunoprotective agents and as vaccine adjuvants. Previous studies of nonhuman primates showed that administration of CpG ODN type D (also known as type A) at the site of infection 3 days before and after a challenge with Leishmania major enhanced host resistance and reduced the lesion's severity. In this study, we show that systemic administration of D/A ODN limits the size of lesions following an intradermal infection with L. major. Importantly, the reduced morbidity was not associated with a reduction in long-term immunity, as such treated macaques were still protected following a secondary challenge. Finally, administration of D/A ODN to macaques that had established cutaneous lesions reduced the severity of the lesions, suggesting a potential role for CpG ODN in L. major treatment. Together, these findings support the development of clinical studies to assess the use of CpG ODN types D/A as immunoprotective and therapeutic agents. PMID- 16041010 TI - Impaired neutrophil migration associated with specific bovine CXCR2 genotypes. AB - Bovine mastitis continues to be the most detrimental factor for profitable dairying. Recent research conducted within our laboratory has identified a genetic marker in the CXCR2 gene associated with mastitis susceptibility. The objective of the present study was to evaluate the migratory ability of neutrophils from cows with different CXCR2 +777 genotypes. Neutrophils isolated from peripheral blood of 30 Holstein cows were tested for in vitro migration and adhesion molecule expression. Cows with the CC or GC genotype at CXCR2 +777 showed significantly lower neutrophil migration to recombinant human interleukin 8 (rhIL-8) than cows with the GG genotype (P < 0.05). Cows with the CC genotype at CXCR2 +777 also showed decreased neutrophil migration to zymosan-activated serum compared to these same cows (P < 0.05). Decreased upregulation of CD18 expression was observed after stimulation with rhIL-8 in cows expressing the CXCR2 +777 CC genotype compared to cows expressing the GG genotype (P < 0.05). A similar trend was observed for CD11b (P < 0.10). However, no difference in CD62 downregulation was observed with respect to genotype. These results provide initial evidence for a phenotypic association between a single nucleotide polymorphism and neutrophil function in dairy cows, as well as potential insight into specific mechanisms affected in cows more susceptible to mastitis. PMID- 16041011 TI - Characterization of the filamentous hemagglutinin-like protein FhaS in Bordetella bronchiseptica. AB - Filamentous hemagglutinin (FHA) is a large (>200 kDa), rod-shaped protein expressed by bordetellae that is both surface-associated and secreted. FHA mediates bacterial adherence to epithelial cells and macrophages in vitro and is absolutely required for tracheal colonization in vivo. The recently sequenced Bordetella bronchiseptica genome revealed the presence of a gene, fhaS, that is nearly identical to fhaB, the FHA structural gene. We show that although fhaS expression requires the BvgAS virulence control system, it is maximal only under a subset of conditions in which BvgAS is active, suggesting an additional level of regulation. We also show that, like FHA, FhaS undergoes a C-terminal proteolytic processing event and is both surface-associated and secreted and that export across the outer membrane requires the channel-forming protein FhaC. Unlike FHA, however, FhaS was unable to mediate adherence of B. bronchiseptica to epithelial cell lines in vitro and was not required for respiratory tract colonization in vivo. In a coinfection experiment, a DeltafhaS strain was out competed by wild-type B. bronchiseptica, indicating that fhaS is expressed in vivo and that FhaS contributes to bacterial fitness in a manner revealed when the mutant must compete with wild-type bacteria. These data suggest that FHA and FhaS perform distinct functions during the Bordetella infectious cycle. A survey of various Bordetella strains revealed two distinct fhaS alleles that segregate according to pathogen host range and that B. parapertussis(hu) most likely acquired its fhaS allele from B. pertussis horizontally, suggesting fhaS may contribute to host-species specificity. PMID- 16041012 TI - Borrelia burgdorferi rel is responsible for generation of guanosine-3' diphosphate-5'-triphosphate and growth control. AB - The global transcriptional regulator (p)ppGpp (guanosine-3'-diphosphate-5' triphosphate and guanosine-3',5'-bisphosphate, collectively) produced by the relA and spoT genes in Escherichia coli allows bacteria to adapt to different environmental stresses. The genome of Borrelia burgdorferi encodes a single chromosomal rel gene (BB0198) (B. burgdorferi rel [rel(Bbu)]) homologous to relA and spoT of E. coli. Its role in (p)ppGpp synthesis, bacterial growth, and modulation of gene expression has not been studied in detail. We constructed a rel(Bbu) deletion mutant in an infectious B. burgdorferi 297 strain and isolated an extrachromosomally complemented derivative of this mutant. The mutant did not synthesize rel(Bbu) mRNA, Rel(Bbu) protein, or (p)ppGpp. This synthesis was restored in the complemented derivative, confirming that rel(Bbu) is necessary and sufficient for (p)ppGpp synthesis and degradation in B. burgdorferi. The rel(Bbu) mutant grew well during log phase in complete BSK-H but reached lower cell concentrations in the stationary phase than the wild-type parent, suggesting that (p)ppGpp may be an important factor in the ability of B. burgdorferi to adapt to stationary phase. Deletion of rel(Bbu) did not eliminate the temperature elicited OspC shift, nor did it alter bmp gene expression or B. burgdorferi antibiotic susceptibility. Although deletion of rel(Bbu) eliminated B. burgdorferi virulence for mice, which was not restored by complementation, we suggest that rel(Bbu)-dependent accumulation of (p)ppGpp may be important for in vivo survival of this pathogen. PMID- 16041013 TI - Characterization of the cciIR quorum-sensing system in Burkholderia cenocepacia. AB - Several transmissible Burkholderia cenocepacia strains that infect multiple cystic fibrosis patients contain a genomic island designated as the cenocepacia island (cci). The cci contains a predicted N-acylhomoserine lactone (AHL) synthase gene, cciI, and a predicted response regulator gene, cciR. AHL production profiles indicated that CciI catalyzes the synthesis of N-hexanoyl-l homoserine lactone and minor amounts of N-octanoyl-l-homoserine lactone. The cciI and cciR genes were found to be cotranscribed by reverse transcription-PCR analysis, and the expression of a cciIR::luxCDABE fusion in a cciR mutant suggested that the cciIR system negatively regulates its own expression. B. cenocepacia strains also have a cepIR quorum-sensing system. Expression of cepI::luxCDABE or cepR::luxCDABE fusions in a cciR mutant showed that CciR negatively regulates cepI but does not regulate cepR. Expression of the cciIR::luxCDABE fusion in a cepR mutant indicated that functional CepR is required for cciIR expression. Phylogenetic analysis suggested that the cciIR system was acquired by horizontal gene transfer from a distantly related organism and subsequently incorporated into the ancestral cepIR regulatory network. Mutations in cciI, cciR, cepI cciI, and cepR cciR were constructed in B. cenocepacia K56-2. The cciI mutant had greater protease activity and less swarming motility than the parent strain. The cciR mutant had less protease activity than the parent strain. The phenotypes of the cepI cciI and cepR cciR mutants were similar to cepI or cepR mutants, with less protease activity and swarming motility than the parent strain. PMID- 16041014 TI - Characterization of enterohemorrhagic Escherichia coli strains based on acid resistance phenotypes. AB - Acid resistance is perceived to be an important property of enterohemorrhagic Escherichia coli strains, enabling the organisms to survive passage through the acidic environment of the stomach so that they may colonize the mammalian gastrointestinal tract and cause disease. Accordingly, the organism has developed at least three genetically and physiologically distinct acid resistance systems which provide different levels of protection. The glutamate-dependent acid resistance (GDAR) system utilizes extracellular glutamate to protect cells during extreme acid challenges and is believed to provide the highest protection from stomach acidity. In this study, the GDAR system of 82 pathogenic E. coli isolates from 34 countries and 23 states within the United States was examined. Twenty nine isolates were found to be defective in inducing GDAR under aerobic growth conditions, while five other isolates were defective in GDAR under aerobic, as well as fermentative, growth conditions. We introduced rpoS on a low-copy-number plasmid into 26 isolates and were able to restore GDAR in 20 acid-sensitive isolates under aerobic growth conditions. Four isolates were found to be defective in the newly discovered LuxR-like regulator GadE (formerly YhiE). Defects in other isolates could be due to a mutation(s) in a gene(s) with an as yet undefined role in acid resistance since GadE and/or RpoS could not restore acid resistance. These results show that in addition to mutant alleles of rpoS, mutations in gadE exist in natural populations of pathogenic E. coli. Such mutations most likely alter the infectivity of individual isolates and may play a significant role in determining the infective dose of enterohemorrhagic E. coli. PMID- 16041015 TI - Immunogenicity of the Mycobacterium tuberculosis PPE55 (Rv3347c) protein during incipient and clinical tuberculosis. AB - Clinical tuberculosis (TB), whether noncavitary or cavitary, is the late stage of a chronic disease process, since Mycobacterium tuberculosis is a slowly growing organism. Our studies have shown that the profiles of antigenic proteins expressed by the in vivo bacteria that elicit antibodies differ in cavitary and noncavitary TB. To gain insight into antigenic proteins expressed during incipient, subclinical TB, an expression library of M. tuberculosis genomic DNA was screened with sera obtained during subclinical TB from guinea pigs infected with aerosols of M. tuberculosis H37Rv. One of the proteins recognized by antibodies elicited during subclinical TB infection of guinea pigs is the 309-kDa PPE55 (Rv3347c) protein. Genomic hybridization studies suggest that the PPE55 gene is specific to the M. tuberculosis complex and is present in a majority of clinical isolates tested. Antibodies to the C-terminal, approximately 100-kDa fragment of PPE55 (PPE-C) were detectable in sera from 29/30 (97%) human immunodeficiency virus-negative/TB-positive (HIV(-) TB(+)) patients and 17/24 (71%) HIV(+) TB(+) patients tested but not in sera from purified-protein derivative-positive healthy controls, suggesting that the in vivo expression of PPE55 protein correlates with active M. tuberculosis infection. Anti-PPE-C antibodies were also detected in retrospective sera obtained months prior to manifestation of clinical TB from 17/21 (81%) HIV(+) TB(+) individuals tested, providing evidence that the protein is expressed during incipient, subclinical TB in HIV-infected humans. Thus, PPE55 is a highly immunogenic protein that may be useful for differentiating between latent TB and incipient, subclinical TB. PMID- 16041016 TI - Endocarditis-associated oral streptococci promote rapid differentiation of monocytes into mature dendritic cells. AB - Endocarditis is frequently attributable to oral streptococci, but mechanisms of pathogenesis are not well understood, although monocytes appear to be important. High titers of interleukin-12 (IL-12) are produced by peripheral blood mononuclear cells (PBMC) after engaging Streptococcus mutans, but monocytes in developing endocardial vegetations tend to disappear rather than become macrophages. These data prompted the hypothesis that streptococcus-infected monocytes differentiate into short-lived IL-12-producing dendritic cells (DCs) rather than macrophages. PBMC from healthy subjects were stimulated with six isolates of oral streptococci, three nonstreptococcal oral bacteria, or IL-4 plus granulocyte-macrophage colony-stimulating factor, and the appearance of cells with markers typical of mature DCs (CD83(+), CD86(+), CD11c(+), and CD14(-)) was monitored. Supernatant fluids from the PBMC cultures were harvested and IL-12 p70 levels were determined. S. mutans-stimulated monocytes were analyzed for their ability to elicit allogeneic mixed-lymphocyte reactions. All streptococci examined, except one strain of Streptococcus oralis (35037), rapidly induced up regulation of CD83 and CD86 and a loss of CD14 in the CD11c(+) monocyte population within 20 h. Induction of IL-12 was CD14 dependent and correlated with streptococcal isolates that promoted the DC phenotype. Major histocompatibility complex (MHC) class II expression was up-regulated by S. mutans, and these cells were short-lived and elicited potent allogeneic mixed-lymphocyte reactions typical of DCs. In summary, monocytes stimulated with endocarditis-associated oral streptococci rapidly exhibited the DC phenotype and functions. These data suggest that the initiation of bacterial endocarditis by oral streptococci may involve monocyte-to-DC differentiation, and this may help explain the low levels of macrophages in the site. PMID- 16041017 TI - Identification and characterization of an SKN7 homologue in Cryptococcus neoformans. AB - Cryptococcus neoformans is an encapsulated fungal pathogen that primarily infects the central nervous system of immunocompromised individuals, causing life threatening meningoencephalitis. The capacity of C. neoformans to subvert host defenses and disseminate by intracellular parasitism of alveolar macrophages in the immune-compromised host has led to studies to evaluate genes associated with C. neoformans resistance to oxidative stress. In the present study, we identify and characterize a C. neoformans homologue to SKN7, a transcription factor in Saccharomyces cerevisiae that regulates the oxidative stress response, cell cycle, and cell wall biosynthesis. To examine the contribution of SKN7 in the pathogenesis of fungal infections, we created skn7 mutants via targeted disruption. The skn7 mutants were observed to be more susceptible to reactive oxygen species in vitro and were significantly less virulent than the wild-type strain and a reconstituted strain as measured by cumulative survival in the mouse inhalational model. The Skn7 protein was observed to be important for expression of thioredoxin reductase in response to oxidative challenge. Interestingly, skn7 mutants were also observed to flocculate following in vitro culture, a novel phenotype not observed in skn7 mutants derived from other fungi. These findings demonstrate that SKN7 contributes to the virulence composite but is not required for pathogenicity in C. neoformans. In addition, flocculation of C. neoformans skn7 mutants suggests a potentially unique function of SKN7 not previously observed in other cryptococcal strains or skn7 mutants. PMID- 16041018 TI - Intranasal delivery of group B meningococcal native outer membrane vesicle vaccine induces local mucosal and serum bactericidal antibody responses in rabbits. AB - We have previously shown that intranasal immunization of mice with meningococcal native outer membrane vesicles (NOMV) induces both a good local mucosal antibody response and a good systemic bactericidal antibody response. However, in the intranasal mouse model, some of the NOMV entered the lung and caused an acute granulocytic response. We therefore developed an alternate animal model using the rabbit. This model reduces the probability of lung involvement and more closely mimics intranasal immunization of humans. Rabbits immunized intranasally with doses of 100 mug of NOMV in 0.5 ml of saline developed serum bactericidal antibody levels comparable to those of rabbits immunized intramuscularly with 25 mug doses, particularly when the primary intranasal immunization was given daily for 3 days. Intranasal immunization also induced a local mucosal response as evidenced by immunoglobulin A antibody in saliva, nasal washes, and lung lavage fluids. NOMV from a capsule-deficient mutant induced higher serum bactericidal antibody responses than NOMV from the encapsulated parent. Meningococcal NOMV could be administered intranasally at 400 mug with no pyrogenic activity, but as little as 0.03 mug/kg of body weight administered intravenously or 25 mug administered intramuscularly induced a pyrogenic response. These data indicate that the rabbit is a useful model for preclinical testing of intranasal meningococcal NOMV vaccines, and this immunization regimen produces a safe and substantial systemic and local mucosal antibody response. PMID- 16041019 TI - Scrub typhus vaccine candidate Kp r56 induces humoral and cellular immune responses in cynomolgus monkeys. AB - A truncated recombinant 56-kDa outer membrane protein of the Karp strain of Orientia tsutsugamushi (Kp r56) was evaluated in cynomolgus monkeys (Macaca fascicularis) for immunogenicity and safety as a vaccine candidate for the prevention of scrub typhus. This recombinant antigen induced strong humoral and cellular immune responses in two monkeys and was found to be well tolerated. Antigen-specific immunoglobulin M (IgM) and IgG were produced to almost maximal levels within 1 week of a single immunization. Peripheral blood mononuclear cells from vaccinated animals showed an induction of antigen-specific proliferation and gamma interferon production. The Kp r56 was not as efficient as infection with live organisms in preventing reinfection but was able to reduce the inflammation produced at the site of challenge. This report describes the results of the first systematic study of the immunogenicity of a recombinant scrub typhus vaccine candidate in a nonhuman primate model. PMID- 16041020 TI - Experimental infection with Trypanosoma cruzi increases the population of CD8(+), but not CD4(+), immunoglobulin G Fc receptor-positive T lymphocytes. AB - It is well established that activating-type Fc receptors for immunoglobulin G (FcgammaR), such as FcgammaRI and FcgammaRIII, are essential for inducing inflammatory responses. On the other hand, a unique inhibitory FcgammaR, FcgammaRIIB, inhibits intracellular signaling upon engagement of immunoglobulin G immune complexes, suppressing inflammation and autoimmunity. The expression of FcgammaRIIB on B lymphocytes, natural killer cells, macrophages, mast cells, and a number of other cell types has been demonstrated for many years. However, the expression on T lymphocytes is probably restricted to activated cells in a narrow window of time. The controversy regarding the FcgammaR expression on T lymphocytes is attributable to considerable heterogeneity of cellular subpopulations and activation stages during immune responses in vivo. We addressed here this question by using mice experimentally infected with Trypanosoma cruzi, and we found an increase in the CD8(+) FcgammaR(+) population but not in the CD4(+) FcgammaR(+) population. Moreover, CD8(+) FcgammaR(+) T cells predominantly composed the cardiac inflammatory infiltration induced by the infection. These results indicate a novel pattern of FcgammaR expression on T cells in a pathological situation, and possible functional roles of this phenomenon are discussed. PMID- 16041021 TI - Identification of a novel cathelicidin gene in the rainbow trout, Oncorhynchus mykiss. AB - We report the cloning of a novel antimicrobial peptide gene, termed rtCATH_1, found in the rainbow trout, Oncorhynchus mykiss. The predicted 216-residue rtCATH_1 prepropeptide consists of three domains: a 22-residue signal peptide, a 128-residue cathelin-like region containing two identifiable cathelicidin family signatures, and a predicted 66-residue C-terminal cationic antimicrobial peptide. This predicted mature peptide was unique in possessing features of different known (mammalian) cathelicidin subgroups, such as the cysteine-bridged family and the specific amino-acid-rich family. The rtCATH_1 gene comprises four exons, as seen in all known mammalian cathelicidin genes, and several transcription factor binding sites known to be of relevance to host defenses were identified in the 5' flanking region. By Northern blot analysis, the expression of rtCATH_1 was detected in gill, head kidney, and spleen of bacterially challenged fish. Primary cultures of head kidney leukocytes from rainbow trout stimulated with lipopolysaccharide or poly(I x C) also expressed rtCATH_1. A 36-residue peptide corresponding to the core part of the fish cathelicidin was chemically synthesized and shown to exhibit potent antimicrobial activity and a low hemolytic effect. Thus, rtCATH_1 represents a novel antimicrobial peptide gene belonging to the cathelicidin family and may play an important role in the innate immunity of rainbow trout. PMID- 16041022 TI - Conditional lethality yields a new vaccine strain of Listeria monocytogenes for the induction of cell-mediated immunity. AB - Listeria monocytogenes is a gram-positive intracellular pathogen that can enter phagocytic and nonphagocytic cells and colonize their cytosols. Taking advantage of this property to generate an intracellular vaccine delivery vector, we previously described a mutant strain of L. monocytogenes, Deltadal Deltadat, which is unable to synthesize cell wall by virtue of deletions in two genes (dal and dat) required for d-alanine synthesis. This highly attenuated strain induced long-lived protective systemic and mucosal immune responses in mice when administered in the transient presence of d-alanine. We have now increased the usefulness of this organism as a vaccine vector by use of an inducible complementation system that obviates the need for exogenous d-alanine administration. The strain expresses a copy of the Bacillus subtilis racemase gene under the control of a tightly regulated isopropyl-beta-d thiogalactopyranoside (IPTG)-inducible promoter present on a multicopy plasmid. This bacterium demonstrates strict dose-dependent growth in the presence of IPTG. After removal of inducer, bacterial growth ceased within two replication cycles. Following infection of mice in the absence of IPTG or d-alanine, the bacterium survived in vivo for less than 3 days. Nevertheless, a single immunization elicited a state of long-lasting protective immunity against wild-type L. monocytogenes and induced a subset of effector listeriolysin O-specific CD11a(+) CD8(+) T cells in spleen and other tissues that was strongly enhanced after secondary immunization. This improved L. monocytogenes vector system may have potential use as a live vaccine against human immunodeficiency virus, other infectious diseases, and cancer. PMID- 16041023 TI - In vitro cellular immune responses to recombinant antigens of Mycobacterium avium subsp. paratuberculosis. AB - Five recombinant antigens (Ags; 85A, 85B, 85C, superoxide dismutase [SOD], and 35 kDa protein) were purified from Mycobacterium avium subsp. paratuberculosis and evaluated for their ability to stimulate peripheral blood mononuclear cells (PMBCs) from fecal-culture-positive cows (low and medium shedders) and culture negative healthy cows. Recombinant Ags 85A, 85B, and 85C induced significant lymphocyte proliferation as well as the production of gamma interferon (IFN gamma), interleukin-2 (IL-2), IL-12, and tumor necrosis factor alpha (TNF-alpha), but not IL-4, from low and medium shedders. The 85 antigen complex did not stimulate PMBC proliferation from culture-negative healthy cows. The 35-kDa protein also induced significant lymphocyte proliferation as well as the production of IFN-gamma and IL-4 from low and medium shedders. CD4(+) T cells and CD25(+) (IL-2R) T cells were stimulated the most by 85A and 85B, while the 35-kDa protein primarily stimulated CD21(+) B cells involved in humoral immune responses. Interestingly, SOD was less immunostimulatory than other antigens but strongly induced gammadelta(+) T cells, which are thought to be important in the early stages of infection, such as pathogen entry. These data provide important insight into how improved vaccines against mycobacterial infections might be constructed. PMID- 16041024 TI - Heme transfer from streptococcal cell surface protein Shp to HtsA of transporter HtsABC. AB - Human pathogen group A streptococcus (GAS) can take up heme from host heme containing proteins as a source of iron. Little is known about the heme acquisition mechanism in GAS. We recently identified a streptococcal cell surface protein (designated Shp) and the lipoprotein component (designated HtsA) of an ATP-binding cassette (ABC) transporter made by GAS as heme-binding proteins. In an effort to delineate the molecular mechanism involved in heme acquisition by GAS, heme-free Shp (apo-Shp) and HtsA (apo-HtsA) were used to investigate heme transfer from heme-containing proteins (holo proteins) to the apo proteins. In addition, the interaction between holo-Shp and holo-HtsA was examined using native polyacrylamide gel electrophoresis. Heme was efficiently transferred from holo-Shp to apo-HtsA but not from holo-HtsA to apo-Shp. Apo-Shp acquired heme from human hemoglobin, and holo-Shp and holo-HtsA were able to form a complex, suggesting that Shp actively relays heme from hemoglobin to apo-HtsA. These findings demonstrate for the first time complex formation and directional heme transfer between a cell surface heme-binding protein and the lipoprotein of a heme-specific ABC transporter in gram-positive bacteria. PMID- 16041025 TI - Identification and characterization of Neospora caninum cyclophilin that elicits gamma interferon production. AB - Gamma interferon (IFN-gamma) response is essential to the development of a host protective immunity in response to infections by intracellular parasites. Neosporosis, an infection caused by the intracellular protozoan parasite Neospora caninum, is fatal when there is a complete lack of IFN-gamma in the infected host. However, the mechanism by which IFN-gamma is elicited by the invading parasite is unclear. This study has identified a microbial protein in the N. caninum tachyzoite N. caninum cyclophilin (NcCyP) as a major component of the parasite responsible for the induction of IFN-gamma production by bovine peripheral blood mononuclear cells (PBMC) and antigen-specific CD4(+) T cells. NcCyP has high sequence homology (86%) with Toxoplasma gondii 18-kDa CyP with a calculated molecular mass of 19.4 kDa. NcCyP is a secretory protein with a predicted signal peptide of 17 amino acids. Abundant NcCyP was detected in whole cell N. caninum tachyzoite lysate antigen (NcAg) and N. caninum tachyzoite culture supernatant. In N. caninum tachyzoite culture supernatant, three NcCyP bands of 19, 22, and 24 kDa were identified. NcAg stimulated high levels of IFN gamma production by PBMC and CD4(+) T cells. The IFN-gamma-inducing effect of NcAg was blocked by cyclosporine, a specific ligand for CyP, in a dose-dependent manner. Furthermore, cyclosporine abolished IFN-gamma production by PBMC from naive cows as well as PBMC and CD4(+) T cells from infected/immunized cows. These results indicate that the N. caninum tachyzoite naturally produces a potent IFN gamma-inducing protein, NcCyP, which may be important for parasite survival as well as host protection. PMID- 16041026 TI - Inter-alpha-inhibitor proteins are endogenous furin inhibitors and provide protection against experimental anthrax intoxication. AB - Inter-alpha-inhibitor protein (IalphaIp) functions as an endogenous serine protease inhibitor in human plasma, and IalphaIp levels diminish rapidly during acute inflammatory states. One potential target for IalphaIp is furin, a cell associated serine endopeptidase essential for the activation of protective antigen and the formation of anthrax lethal toxin (LT). IalphaIp blocks furin activity in vitro and provides significant protection against cytotoxicity for murine peritoneal macrophages exposed to up to 500 ng/ml LT. A monoclonal antibody (MAb), 69.31, that specifically blocks the enzymatic activity of IalphaIp eliminates its protective effect against LT-induced cytotoxicity. IalphaIp (30 mg/kg of body weight) administered to BALB/c mice 1 hour prior to an intravenous LT challenge resulted in 71% survival after 7 days compared with no survivors among the control animals (P < 0.001). We conclude that human IalphaIp may be an effective preventative or therapeutic agent against anthrax intoxication. PMID- 16041027 TI - Establishment of cloned Anaplasma phagocytophilum and analysis of p44 gene conversion within an infected horse and infected SCID mice. AB - Diverse p44 alleles at the p44 expression locus (p44Es) encoding surface-exposed major membrane proteins, P44s, of Anaplasma phagocytophilum were hypothesized to be garnered by recombination to enact antigenic variation. However, this hypothesis has not been proven so far, due to inability to clone this obligate intragranulocytic rickettsia. To define the p44E recombination, we developed a novel method to clone A. phagocytophilum. This isogenic cloned population containing a defined p44E was used to infect a naive horse and severe combined immunodeficiency (SCID) mice. During a 58-day infection period in the blood of the horse, p44E conversion was evident in a total of 11 new p44Es, 48% (115/242) of the sequenced p44E population. During a 50-day infection period in the blood of SCID mice, p44E conversion was manifested in a total of 13 new p44Es, 42% (192/460) of the p44E population. Thus, similar levels of p44E convertants were detected in either the presence or absence of an acquired immune system, suggesting that T- and B-cell immune pressure was not essential for recombination and/or selection of the p44E variants. Analysis of sequentially changed p44Es revealed that the entire central hypervariable region of donor p44 pseudogenes or of donor full-length p44s replaced the same region of the resident p44E as a cassette. Putative recombination points were detected within p44 conserved regions flanking the central hypervariable region by the TOPALi analysis. Our results unambiguously demonstrated p44E recombination. The cloning method developed would facilitate precise analysis of the recombination process and the extent of diversity which the recombination creates in the antigenic repertoire. PMID- 16041028 TI - Shiga toxin 1 induces apoptosis in the human myelogenous leukemia cell line THP-1 by a caspase-8-dependent, tumor necrosis factor receptor-independent mechanism. AB - Shiga toxins (Stxs) induce apoptosis in a variety of cell types. Here, we show that Stx1 induces apoptosis in the undifferentiated myelogenous leukemia cell line THP-1 in the absence of tumor necrosis factor alpha (TNF-alpha) or death receptor (TNF receptor or Fas) expression. Caspase-8 and -3 inhibitors blocked, and caspase-6 and -9 inhibitors partially blocked, Stx1-induced apoptosis. Stx1 induced the mitochondrial pathway of apoptosis, as activation of caspase-8 triggered the (i) cleavage of Bid, (ii) disruption of mitochondrial membrane potential, and (iii) release of cytochrome c into the cytoplasm. Caspase-8, -9, and -3 cleavage and functional activities began 4 h after toxin exposure and peaked after 8 h of treatment. Caspase-6 may also contribute to Stx1-induced apoptosis by directly acting on caspase-8. It appears that functional Stx1 holotoxins must be transported to the endoplasmic reticulum to initiate apoptotic signaling through the ribotoxic stress response. These data suggest that Stxs may activate monocyte apoptosis via a novel caspase-8-dependent, death receptor independent mechanism. PMID- 16041029 TI - Hag directly mediates the adherence of Moraxella catarrhalis to human middle ear cells. AB - Moraxella catarrhalis is a human pathogen that causes otitis media in young children and lung infections in patients with chronic obstructive pulmonary disease. In this study, the role of the surface protein Hag in the adherence of multiple M. catarrhalis strains was examined. The hag genes of four clinical isolates were disrupted with a spectinomycin resistance cassette, and the binding of isogenic mutants to primary cultures of human middle ear epithelial cells (HMEE), as well as A549 pneumocytes, was measured. These experiments revealed that the attachment of most mutants to both cell types was 10-fold less than that of their wild-type progenitors. To determine whether Hag directly mediates adherence to human cells, the hag genes from three M. catarrhalis isolates were cloned and expressed in a nonadherent Escherichia coli cloning strain. At least 17-fold more E. coli bacteria expressing Hag attached to HMEE cells than an adherence-negative control. Surprisingly, Hag expression did not increase the binding of recombinant E. coli to A549 monolayers. Our data demonstrate that the involvement of Hag in M. catarrhalis adherence to A549 and HMEE cells is conserved among isolates and that Hag directly mediates binding to HMEE cells. PMID- 16041030 TI - MyD88, but not toll-like receptors 4 and 2, is required for efficient clearance of Brucella abortus. AB - It is not clear how the host initially recognizes and responds to infection by gram-negative pathogenic Brucella spp. It was previously shown (D. S. Weiss, B. Raupach, K. Takeda, S. Akira, and A. Zychlinsky, J. Immunol. 172:4463-4469, 2004) that the early macrophage response against gram-negative bacteria is mediated by Toll-like receptor 4 (TLR4), which signals in response to lipopolysaccharide (LPS). Brucella, however, has a noncanonical LPS which does not have potent immunostimulatory activity. We evaluated the kinetics of TLR4 activation and the cytokine response in murine macrophages after Brucella infection. We found that during infection of macrophages, Brucella avoids activation of TLR4 at 6 h but activates TLR4, TLR2, and myeloid differentiation factor 88 (MyD88) at 24 h postinfection. Interestingly, even though its activation is delayed, MyD88 is important for host defense against Brucella infection in vivo, since MyD88(-/-) mice do not clear the bacteria as efficiently as wild-type, TLR4(-/-), TLR2(-/-), or TLR4/TLR2(-/-) mice. PMID- 16041031 TI - 4-1BB (CD137) is required for rapid clearance of Listeria monocytogenes infection. AB - 4-1BB (CD137), a member of the tumor necrosis factor receptor superfamily, is a T cell-costimulatory receptor that is expressed on activated T cells, dendritic cells, and NK cells. Little has been reported about its role in early host defense against bacterial infection. In this study, we report that 4-1BB deficient (4-1BB(-/-)) mice are much more susceptible to Listeria monocytogenes (intracellular bacteria) infections than wild-type mice. Upon L. monocytogenes infection, 4-1BB(-/-) mice showed a lower survival rate, a higher bacterial burden in organs, and larger hepatic microabscesses than 4-1BB(+/+) mice. 4-1BB( /-) mice also had impairment in clearance of bacteria from the bloodstream. Neutrophils from 4-1BB(+/+) mice constitutively expressed 4-1BB, which could be activated to induce intracellular Ca(2+) influx by ligation with anti-4-1BB antibody. On the other hand, neutrophils from 4-1BB(-/-) mice were defective in reactive oxygen species generation, phagocytic activities, and intracellular Ca(2+) mobilization. In addition, mice pretreated with anti-4-1BB monoclonal antibody were much more resistant to L. monocytogenes infection than control antibody-treated mice. Our results support the notion that 4-1BB may play a major role in host defense against intracellular pathogens through neutrophil activation. PMID- 16041032 TI - LcrV plague vaccine with altered immunomodulatory properties. AB - Yersinia pestis, the causative agent of plague, secretes LcrV (low-calcium response V or V antigen) during infection. LcrV triggers the release of interleukin 10 (IL-10) by host immune cells and suppresses proinflammatory cytokines such as tumor necrosis factor alpha and gamma interferon as well as innate defense mechanisms required to combat the pathogenesis of plague. Although immunization of animals with LcrV elicits protective immunity, the associated suppression of host defense mechanisms may preclude the use of LcrV as a human vaccine. Here we show that short deletions within LcrV can reduce its immune modulatory properties. An LcrV variant lacking amino acid residues 271 to 300 (rV10) elicited immune responses that protected mice against a lethal challenge with Y. pestis. Compared to full-length LcrV, rV10 displayed a reduced ability to release IL-10 from mouse and human macrophages. Furthermore, the lipopolysaccharide-stimulated release of proinflammatory cytokines by human or mouse macrophages was inhibited by full-length LcrV but not by the rV10 variant. Thus, it appears that LcrV variants with reduced immune modulatory properties could be used as a human vaccine to generate protective immunity against plague. PMID- 16041033 TI - Experimental ocular toxoplasmosis in genetically susceptible and resistant mice. AB - Genetic factors determining the pathogenesis and course of ocular toxoplasmosis are poorly understood. In this study, we explored the development of experimental ocular pathogenesis in genetically dissimilar mice infected with either the RH strain, the PLK strain, or the immunodominant surface antigen 1 (SAG1 [P30]) deficient mutant of the RH strain of Toxoplasma gondii. At 11 days postinfection, ocular infection of C57BL/6 mice with all of the strains of parasites resulted in severe inflammatory lesions and high numbers of parasites in eye tissue; less severe ocular lesions at earlier histopathology and prolonged survival were observed in this mouse strain infected with either the major surface antigen 1 deficient SAG1(-/-) strain or the less virulent PLK strain compared with RH infection. In contrast, both BALB/c and CBA/J mice had less severe lesions and low numbers of parasites in their eye tissue, and infection developed into the chronic stage in these mice. There were significantly higher serum levels of gamma interferon and tumor necrosis factor alpha in C57BL/6 mice than in BALB/c and CBA/J mice following ocular infection. These observations confirm earlier reports on systemic immunity to these parasites that the route of Toxoplasma infection markedly influences survival of mice. Our data indicate that genetic factors of the host as well as the parasite strain are critical in determining susceptibility to experimental ocular toxoplasmosis in murine models. PMID- 16041034 TI - Production and characterization of monoclonal antibodies against Enterocytozoon bieneusi purified from rhesus macaques. AB - Enterocytozoon bieneusi spores derived from rhesus macaque feces were purified by serial salt-Percoll-sucrose-iodixanol centrifugation, resulting in two bands with different specific densities of 95.6% and 99.5% purity and with a recovery efficiency of 10.8%. An ultrastructural examination revealed typical E. bieneusi spores. Twenty-six stable hybridomas were derived from BALB/c mice immunized with spores and were cloned twice by limiting dilution or growth on semisolid medium. Four monoclonal antibodies (MAbs), reacting exclusively with spores, were further characterized. These MAbs specifically reacted with spores present in stools of humans and macaques, as visualized by immunofluorescence, and with spore walls, as visualized by immunoelectron microscopy. A blocking enzyme-linked immunosorbent assay and Western blotting revealed that the epitope recognized by 8E2 was different from those recognized by 7G2, 7H2, and 12G8, which identified the same 40-kDa protein. These MAbs will be valuable tools for diagnostics, for epidemiological investigations, for host-pathogen interaction studies, and for comparative genomics and proteomics. PMID- 16041035 TI - Cytokine and chemokine responses associated with clearance of a primary Salmonella enterica serovar Typhimurium infection in the chicken and in protective immunity to rechallenge. AB - Infection of poultry with Salmonella enterica serovar Typhimurium poses a significant risk to public health through contamination of meat from infected animals. Vaccination has been proposed to control infections in chickens. However, the vaccines are currently largely empirical, and our understanding of the mechanisms that underpin immune clearance and protection in avian salmonellosis is not complete. In this study we describe the cytokine, chemokine, and antibody responses and cellular changes in primary and secondary infections of chickens with Salmonella serovar Typhimurium. Infection of 1-week-old chickens induced early expression of a macrophage inflammatory protein (MIP) family chemokine in the spleen and liver, followed by increased expression of gamma interferon accompanied by increased numbers of both CD4(+) and CD8(+) T cells and the formation of granuloma-like follicular lesions. This response correlated with a Th1-mediated clearance of the systemic infection. Primary infection also induced specific immunoglobulin M (IgM), IgG, and IgA antibody responses. In contrast to previously published studies performed with newly hatched chicks, the expression levels of proinflammatory cytokines in the gastrointestinal tract were not greatly increased following infection. However, significant expression of the anti-inflammatory cytokine transforming growth factor beta4 was detected in the gut early in infection. Following secondary challenge, the birds were fully protected against systemic infection and showed a high level of protection against gastrointestinal colonization. Rapid expression of the MIP family chemokine and interleukin-6 was detected in the guts of these birds and was accompanied by an influx of lymphocytes. Increased levels of serum IgA-specific antibodies were also found following rechallenge. These findings suggest that cellular responses, particularly Th1 responses, play a crucial role in immune clearance in avian salmonellosis and that protection against rechallenge involves the rapid recruitment of cells to the gastrointestinal tract. Additionally, the high levels of inflammatory response found following Salmonella serovar Typhimurium infection of newly hatched chicks were not observed following infection of older birds (1 week old), in which the expression of regulatory cytokines appeared to limit inflammation. PMID- 16041036 TI - Probiotics reduce enterohemorrhagic Escherichia coli O157:H7- and enteropathogenic E. coli O127:H6-induced changes in polarized T84 epithelial cell monolayers by reducing bacterial adhesion and cytoskeletal rearrangements. AB - The aim of this study was to determine if probiotics reduce epithelial injury following exposure to Escherichia coli O157:H7 and E. coli O127:H6. The pretreatment of intestinal (T84) cells with lactic acid-producing bacteria reduced the pathogen-induced drop in transepithelial electrical resistance. These findings demonstrate that probiotics prevent epithelial injury induced by attaching-effacing bacteria. PMID- 16041037 TI - Factors associated with severe granulomatous pneumonia in Mycobacterium tuberculosis-infected mice vaccinated therapeutically with hsp65 DNA. AB - Resistant C57BL/6 mice infected in the lungs with Mycobacterium tuberculosis and then therapeutically vaccinated with Mycobacterium leprae-derived hsp65 DNA develop severe granulomatous pneumonia and tissue damage. Analysis of cells accumulating in the lungs of these animals revealed substantial increases in T cells secreting tumor necrosis factor alpha and CD8 cells staining positive for granzyme B. Stimulation of lung cells ex vivo revealed very high levels of interleukin-10, some of which was produced by B-1 B cells. This was probably an anti-inflammatory response, since lung pathology was dramatically worsened in B cell gene-disrupted mice. PMID- 16041038 TI - Intracellular survival of Campylobacter jejuni in human monocytic cells and induction of apoptotic death by cytholethal distending toxin. AB - Campylobacter jejuni 81-176 is capable of extensive replication within human monocytic cell vacuoles and induces apoptotic death via cytolethal distending toxin. PMID- 16041039 TI - Biofilm formation by Salmonella enterica serovar Typhimurium and Escherichia coli on epithelial cells following mixed inoculations. AB - Biofilms were formed by inoculations of Salmonella enterica serovar Typhimurium and Escherichia coli on HEp-2 cells. Inoculations of S. enterica serovar Typhimurium and E. coli resulted in the formation of an extensive biofilm of S. enterica serovar Typhimurium. In experiments where an E. coli biofilm was first formed followed by challenge with S. enterica serovar Typhimurium, there was significant biofilm formation by S. enterica serovar Typhimurium. The results of this study indicate that S. enterica serovar Typhimurium can outgrow E. coli in heterologous infections and displace E. coli when it forms a biofilm on HEp-2 cells. PMID- 16041040 TI - Virulence comparisons of Aspergillus nidulans mutants are confounded by the inflammatory response of p47phox-/- mice. AB - While investigating the requirement for phagosomal alkalinization in the host defense against pulmonary aspergillosis, we observed high morbidity of p47(phox)( /-) mice infected with pH-insensitive Aspergillus nidulans mutants despite a paucity of fungal growth. Fatal infection also resulted from a normally avirulent p-aminobenzoate auxotroph. This demonstrates that p47(phox)(-/-) murine immunity contributes significantly to A. nidulans lethality. These data have wider implications for microbial virulence studies with p47(phox)(-/-) mice. PMID- 16041041 TI - Identification of variant-specific surface proteins in Giardia muris trophozoites. AB - Giardia lamblia undergoes antigenic variation, a process that might allow the parasite to evade the host's immune response and adapt to different environments. Here we show that Giardia muris, a related species that naturally infects rodents, possesses multiple variant-specific surface proteins (VSPs) and expresses VSPs on its surface, suggesting that it undergoes antigenic variation similar to that of G. lamblia. PMID- 16041042 TI - Staphylococcus aureus peptidoglycan is a toll-like receptor 2 activator: a reevaluation. AB - Since the ability of peptidoglycan (PGN) to activate Toll-like receptor 2 (TLR2) was recently questioned, we reevaluated activation of TLR2 by PGN. Polymeric soluble or insoluble Staphylococcus aureus PGN, repurified by sodium dodecyl sulfate or phenol extraction, activated TLR2 at 0.1 to 1 or 10 mug/ml, respectively, and induced tumor necrosis factor alpha production. The TLR2 activation by PGN, but not by lipoteichoic acid, was abolished by muramidase digestion. We conclude that polymeric S. aureus PGN is a TLR2 activator. PMID- 16041043 TI - Selective capture of Salmonella enterica serovar typhi genes expressed in macrophages that are absent from the Salmonella enterica serovar Typhimurium genome. AB - Thirty-six Salmonella enterica serovar Typhi-specific genes, absent from the Salmonella enterica serovar Typhimurium genome, that were expressed in human macrophages were identified by selective capture of transcribed sequences. These genes are located on 15 unique loci of the serovar Typhi genome, including Salmonella pathogenicity islands (SPI-7, SPI-8, and SPI-10) and bacteriophages (ST15, ST18, and ST35). PMID- 16041044 TI - Importance of srtA and srtB for growth of Bacillus anthracis in macrophages. AB - We examined the effect of mutation of two sortase genes of Bacillus anthracis, srtA and srtB, on the ability of the bacterium to grow in J774A.1 cells, a mouse macrophage-like cell line. While disruption of either srtA or srtB had no effect on the ability of the bacteria to grow in rich culture media, mutations in each of these genes dramatically attenuated growth of the bacterium in J774A.1 cells. Complementation of the mutation restored the ability of bacteria to grow in the cells. Since the initial events in inhalation anthrax are believed to be uptake of B. anthracis spores by alveolar macrophages followed by germination of the spores and growth of the bacteria within the macrophages, these results suggest that two sortases of B. anthracis may be critical in the early stages of inhalation anthrax. PMID- 16041045 TI - Characterization of a humanized monoclonal antibody recognizing clumping factor A expressed by Staphylococcus aureus. AB - We report the humanization and characterization of monoclonal antibody (MAb) T1-2 or tefibazumab, a monoclonal antibody that recognizes clumping factor A expressed on the surface of Staphylococcus aureus. We demonstrate that the binding kinetics of MAb T1-2 is indistinguishable compared to that of its murine parent. Furthermore, MAb T1-2 is shown to enhance the opsonophagocytic uptake of ClfA coated latex beads, protect against an intravenous challenge in a prophylactic model of rabbit infective endocarditis, and enhance the efficacy of vancomycin therapy in a therapeutic model of established infective endocarditis. PMID- 16041046 TI - Mapping of the binding site for Mannheimia haemolytica leukotoxin within bovine CD18. AB - To map the site involved in Mannheimia haemolytica leukotoxin (LktA) binding and biological activity within bovine CD18, bovine x human CD18 chimeric constructs were generated and coexpressed with bovine CD11a in K562 cells. Studies with the chimeric leukocyte function-associated antigen 1 transductants demonstrate that the site required for LktA binding and biological effects resides within amino acid residues 500 and 600 of the extracellular region of bovine CD18. PMID- 16041047 TI - Role for mannose binding lectin in the prevention of Mycoplasma infection. AB - Polymorphisms in exon 1 of the MBL-2 gene, resulting in reduced plasma levels of mannose binding lectin, were significantly overrepresented in 23 patients with primary antibody deficiency and culture-proven mycoplasma infections (P = 0.0038). This association persisted with the inclusion of a further nine suspected (doxycycline-responsive) cases (P = 0.0087). The lectin was shown to bind to three strains of mycoplasma. PMID- 16041048 TI - Staphylococcus aureus infection of epidermal keratinocytes promotes expression of innate antimicrobial peptides. AB - Keratinocytes upregulate expression of endogenous antimicrobial peptides in response to inflammatory stimuli. We show that both viable and heat-inactivated Staphylococcus aureus and lipoteichoic acid differentially alter expression of these peptides upon contact with human keratinocytes. The findings indicate a diversity of staphylococcal factors involved in upregulation of antimicrobial peptide expression in cutaneous epithelia. PMID- 16041049 TI - Recombinant proteins of Cryptosporidium parvum induce proliferation of mesenteric lymph node cells in infected mice. AB - Recombinant antigens of Cryptosporidium parvum, Cp900 and Cp40 but not Cp15, stimulated C. parvum-specific proliferative immune responses of mesenteric lymph node cells in C57BL/6J mice infected with different isolates (MD, GCH1, UCP, and IOWA) of C. parvum, indicating that both Cp900 and Cp40 are immunodominant targets of cellular immune responses during C. parvum infection. PMID- 16041050 TI - Prolonged survival of a murine model of cerebral malaria by kynurenine pathway inhibition. AB - C57BL/6J mice infected with Plasmodium berghei ANKA develop neurological dysfunction and die within 7 days of infection. We show that treatment of infected mice with a kynurenine-3-hydroxylase inhibitor prevents them from developing neurological symptoms and extends their life span threefold until severe anemia develops. PMID- 16041051 TI - Infectivity of Cryptosporidium hominis and Cryptosporidium parvum genotype 2 isolates in immunosuppressed Mongolian gerbils. AB - One-month-old dexamethasone-immunosuppressed Mongolian gerbils were challenged with 1 oocyst to 2 x 10(5) oocysts from two isolates genotyped as Cryptosporidium hominis and C. parvum (genotype 2), respectively. A similar dose-dependent gut infection was obtained, and the initial genotype maintained for 21 to 22 days. The data suggest that immunosuppressed gerbils provide a reliable rodent model of persistent C. hominis infection. PMID- 16041052 TI - Protection against aerosolized Yersinia pestis challenge following homologous and heterologous prime-boost with recombinant plague antigens. AB - A Yersinia pestis-derived fusion protein (F1-V) has shown great promise as a protective antigen against aerosol challenge with Y. pestis in murine studies. In the current study, we examined different prime-boost regimens with F1-V and demonstrate that (i) boosting by a route other than the route used for the priming dose (heterologous boosting) protects mice as well as homologous boosting against aerosol challenge with Y. pestis, (ii) parenteral immunization is not required to protect mice against aerosolized plague challenge, (iii) the route of immunization and choice of adjuvant influence the magnitude of the antibody response as well as the immunoglobulin G1 (IgG1)/IgG2a ratio, and (iv) inclusion of an appropriate adjuvant is critical for nonparenteral immunization. PMID- 16041053 TI - Similarity of gene expression patterns in human alveolar macrophages in response to Pseudomonas aeruginosa and Burkholderia cepacia. AB - To determine if differences in the severity of pulmonary infection in cystic fibrosis seen with late isolates of Pseudomonas aeruginosa and Burkholderia cepacia are associated with differences in the initial response of alveolar macrophages (AM) to these pathogens, we assessed gene expression changes in human AM in response to infection with a laboratory strain, early and late clinical isolates of P. aeruginosa, and B. cepacia. Analysis of gene expression changes at the RNA level using oligonucleotide microarrays, following exposure to laboratory P. aeruginosa strain PAK, showed significant (P < 0.01) >2.5-fold upregulation of 42 genes and >2.5-fold downregulation of 45 genes. The majority of the changes in gene expression involved genes as part of inflammatory pathways and signaling systems. Interestingly, similar responses were observed following exposure of AM to early and late clinical isolates of P. aeruginosa, as well as with B. cepacia, suggesting that the more severe clinical outcome of infections with late clinical isolates of P. aeruginosa or with B. cepacia cannot be explained by differences in the early interactions of these organisms with the human AM, as reflected by the similarity of gene expression changes in response to exposure of AM to these pathogens. PMID- 16041054 TI - Investigation of oxidative stress defenses of Neisseria gonorrhoeae by using a human polymorphonuclear leukocyte survival assay. AB - Neisseria gonorrhoeae has well-characterized oxidative stress defense systems that protect against oxidative killing in in vitro assays. In contrast, mutant strains of N. gonorrhoeae lacking oxidative stress defenses are identical to the wild type when tested in an ex vivo survival assay using human polymorphonuclear leukocytes. PMID- 16041055 TI - Mutation of the zinc-binding metalloprotease motif affects Bacteroides fragilis toxin activity but does not affect propeptide processing. AB - To evaluate the role of the zinc-binding metalloprotease in Bacteroides fragilis toxin (BFT) processing and activity, the zinc-binding consensus sequences (H348, E349, H352, G355, H358, and M366) were mutated by site-directed-mutagenesis. Our results indicated that single point mutations in the zinc-binding metalloprotease motif do not affect BFT processing but do reduce or eliminate BFT biologic activity in vitro. PMID- 16041056 TI - Campylobacter jejuni gene expression in the chick cecum: evidence for adaptation to a low-oxygen environment. AB - Transcriptional profiling of Campylobacter jejuni during colonization of the chick cecum identified 59 genes that were differentially expressed in vivo compared with the genes in vitro. The data suggest that C. jejuni regulates electron transport and central metabolic pathways to alter its physiological state during establishment in the chick cecum. PMID- 16041057 TI - Heterologous prime-boost vaccination with the LACK antigen protects against murine visceral leishmaniasis. AB - This study reports the efficacy of a heterologous prime-boost vaccination using DNA and vaccinia viruses (Western Reserve [WR] virus and modified [attenuated] vaccinia virus Ankara [MVA]) expressing the LACK antigen (Leishmania homologue of receptors for activated C kinase) and an intradermal murine infection model employing Leishmania infantum. At 1 month postinfection, vaccinated mice showed high levels of protection in the draining lymph node (240-fold reduction in parasite burden) coupled with significant levels of gamma interferon (20 to 200 ng/ml) and tumor necrosis factor alpha/lymphotoxin (8 to 134 pg/ml). Significant but lower levels of protection (6- to 30-fold) were observed in the spleen and liver. Comparable levels of protection were found for mice boosted with either LACK-WR or LACK-MVA, supporting the use of an attenuated vaccinia virus-based vaccine against human visceral leishmaniasis. PMID- 16041058 TI - Development of an internally controlled antibody microarray. AB - Antibody microarrays are a high throughput technology used to concurrently screen for protein expression. Most antibody arrays currently used are based on the ELISA sandwich approach that uses two antibodies to screen for the expression of a limited number of proteins. Also because antigen-antibody interactions are concentration-dependent, antibody microarrays need to normalize the amount of antibody that is used. In response to the limitations with the currently existing technology we have developed a single antibody-based microarray where the quantity of antibody spotted is used to standardize the antigen concentration. In addition, this new array utilizes an internally controlled system where one color represents the amount of antibody spotted, and the other color represents the amount of the antigen that is used to quantify the level of protein expression. When compared with median fluorescence intensity alone, normalization for antibody spot intensity decreased variability and lowered the limits of detection. This new antibody array was tested using standard cytokine proteins and also cell lysates obtained from mouse macrophages stimulated in vitro and evaluated for the expression of the cytokine proteins interleukin (IL)-1beta, IL 5, IL-6, and macrophage inflammatory proteins 1alpha and 1beta. The levels of protein expression seen with the antibody microarray was compared with that obtained with Western blot analysis, and the magnitude of protein expression observed was similar with both technologies with the antibody array actually showing a greater degree of sensitivity. In summary, we have developed a new type of antibody microarray to screen for protein expression that utilizes a single antibody and controls for the amount of antibody spotted. This type of array appears at least as sensitive as Western blot analysis, and the technology can be scaled up for high throughput screening for hundreds of proteins in complex biofluids such as blood. PMID- 16041059 TI - Functional coupling of cleavage and polyadenylation with transcription of mRNA. AB - Cleavage and polyadenylation define the 3' ends of almost all eukaryotic mRNAs and are thought to occur during transcription. We describe a human in vitro system utilizing an immobilized template, in which transcripts in RNA polymerase II elongation complexes are efficiently cleaved and polyadenylated. Because the cleavage rate of free RNA is much slower, we conclude that cleavage is functionally coupled to transcription. Inhibition of positive transcription elongation factor b (P-TEFb) had only a modest negative effect on cleavage, as long as transcripts were long enough to contain the polyadenylation signal. In contrast, removal of the carboxyl-terminal domain of the large subunit of RNA polymerase II had a dramatic negative effect on cleavage. Unexpectedly, the 5' portion of transcript after cleavage remained associated with the template in a functional, polyadenylation-competent complex. Efficient cleavage required 5' capping by the human capping enzyme, but the reduction of cleavage seen of transcripts in COOH-terminal domain-less polymerase elongation complexes, was not because of lack of capping. PMID- 16041060 TI - Translocation by the RecB motor is an absolute requirement for {chi}-recognition and RecA protein loading by RecBCD enzyme. AB - RecBCD enzyme is a heterotrimeric helicase/nuclease that initiates homologous recombination at double-stranded DNA breaks. The enzyme is driven by two motor subunits, RecB and RecD, translocating on opposite single-strands of the DNA duplex. Here we provide evidence that, although both motor subunits can support the translocation activity for the enzyme, the activity of the RecB subunit is necessary for proper function of the enzyme both in vivo and in vitro. We demonstrate that the RecBCD(K177Q) enzyme, in which RecD helicase is disabled by mutation of the ATPase active site, complements recBCD deletion in vivo and displays all of the enzymatic activities that are characteristic of the wild-type enzyme in vitro. These include helicase and nuclease activities and the abilities to recognize the recombination hotspot chi and to coordinate the loading of RecA protein onto the ssDNA it produces. In contrast, the RecB(K29Q)CD enzyme, carrying a mutation in the ATPase site of RecB helicase, fails to complement recBCD deletion in vivo. We further show that even though RecB(K29Q)CD enzyme displays helicase and nuclease activities, its inability to translocate along the 3'-terminated strand results in the failure to recognize chi and to load RecA protein. Our findings argue that translocation by the RecB motor is required to deliver RecC subunit to chi, whereas the RecD subunit has a dispensable motor activity but an indispensable regulatory function. PMID- 16041061 TI - Bipolar DNA translocation contributes to highly processive DNA unwinding by RecBCD enzyme. AB - We recently demonstrated that the RecBCD enzyme is a bipolar DNA helicase that employs two single-stranded DNA motors of opposite polarity to drive translocation and unwinding of duplex DNA. We hypothesized that this organization may explain the exceptionally high rate and processivity of DNA unwinding catalyzed by the RecBCD enzyme. Using a stopped-flow dye displacement assay for unwinding activity, we test this idea by analyzing mutant RecBCD enzymes in which either of the two helicase motors is inactivated by mutagenesis. Like the wild type RecBCD enzyme, the two mutant proteins maintain the ability to bind tightly to blunt duplex DNA ends in the absence of ATP. However, the rate of forward translocation for the RecB motor-defective enzyme is only approximately 30% of the wild-type rate, whereas for the RecD motor-defective enzyme, it is approximately 50%. More significantly, the processivity of translocation is substantially reduced by approximately 25- and 6-fold for each mutant enzyme, respectively. Despite retaining the capacity to bind blunt dsDNA, the RecB-mutant enzyme has lost the ability to unwind DNA unless the substrate contains a short 5'-terminated single-stranded DNA overhang. The consequences of this observation for the architecture of the single-stranded DNA motors in the initiation complex are discussed. PMID- 16041062 TI - Controlling {beta}-amyloid oligomerization by the use of naphthalene sulfonates: trapping low molecular weight oligomeric species. AB - Aggregation of proteins and peptides has been shown to be responsible for several diseases known as amyloidoses, which include Alzheimer disease (AD), prion diseases, among several others. AD is a neurodegenerative disorder caused primarily by the aggregation of beta-amyloid peptide (Abeta). Here we describe the stabilization of small oligomers of Abeta by the use of sulfonated hydrophobic molecules such as AMNS (1-amino-5-naphthalene sulfonate); 1,8-ANS (1 anilinonaphthalene-8-sulfonate) and bis-ANS (4,4'-dianilino-1,1'-binaphthyl-5,5' disulfonate). The experiments were performed with either Abeta-1-42 or with Abeta 13-23, a shorter version of Abeta that is still able to form amyloid fibrils in vitro and contains amino acid residues 16-20, previously shown to be essential to peptide-peptide interaction and fibril formation. All sulfonated molecules tested were able to prevent Abeta aggregation in a concentration dependent fashion in the following order of efficacy: 1,8-ANS < AMNS < bis-ANS. Size exclusion chromatography revealed that in the presence of bis-ANS, Abeta forms a heterogeneous population of low molecular weight species that proved to be toxic to cell cultures. Since the ANS compounds all have apolar rings and negative charges (sulfonate groups), both hydrophobic and electrostatic interactions may contribute to interpeptide contacts that lead to aggregation. We also performed NMR experiments to investigate the structure of Abeta-13-23 in SDS micelles and found features of an alpha-helix from Lys(16) to Phe(20). 1H TOCSY spectra of Abeta-13-23 in the presence of AMNS displayed a chemical-shift dispersion quite similar to that observed in SDS, which suggests that in the presence of AMNS this peptide might adopt a conformation similar to that reported in the presence of SDS. Taken together, our studies provide evidence for the crucial role of small oligomers and their stabilization by sulfonate hydrophobic compounds. PMID- 16041063 TI - The role of glial cells and apoptosis of enteric neurones in the neuropathology of intractable slow transit constipation. AB - BACKGROUND: Idiopathic slow transit constipation is one of the most severe and often intractable forms of constipation. As motor abnormalities are thought to play an important pathogenetic role, studies have been performed on the colonic neuroenteric system, which rules the motor aspects of the viscus. AIMS: We hypothesised that important neuropathological abnormalities of the large bowel are present, that these are not confined to the interstitial cells of Cajal and ganglion cells, and that the previously described reduction of enteric neurones, if confirmed, might be related to an increase in programmed cell death (apoptosis). PATIENTS AND METHODS: Surgical specimens from 26 severely constipated patients were assessed by conventional and immunohistochemical methods. Specific staining for enteric neurones, glial cells, interstitial cells of Cajal, and fibroblast-like cells associated with the latter were used. In addition, gangliar cell apoptosis was evaluated by means of indirect and direct techniques. Data from patients were compared with those obtained in 10 controls. RESULTS: Severely constipated patients displayed a significant decrease in enteric gangliar cells, glial cells, and interstitial cells of Cajal. Fibroblast like cells associated with the latter did not differ significantly between patients and controls. Patients had significantly more apoptotic enteric neurones than controls. CONCLUSION: Severely constipated patients have important neuroenteric abnormalities, not confined to gangliar cells and interstitial cells of Cajal. The reduction of enteric neurones may in part be due to increased apoptotic phenomena. PMID- 16041064 TI - Dose dependent and divergent effects of superoxide anion on cell death, proliferation, and migration of activated human hepatic stellate cells. AB - BACKGROUND AND AIMS: Activated myofibroblast-like cells, originating from hepatic stellate cells (HSC/MFs) or other cellular sources, play a key profibrogenic role in chronic liver diseases (CLDs) that, as suggested by studies in animal models or rat HSC/MFs, may be modulated by reactive oxygen intermediates (ROI). In this study, human HSC/MFs, exposed to different levels of superoxide anion (O(2)(.-)) and, for comparison, hydrogen peroxide (H(2)O(2)), were analysed in terms of cytotoxicity, proliferative response, and migration. METHODS: Cultured human HSC/MFs were exposed to controlled O(2)(.-) generation by hypoxanthine/xanthine oxidase systems or to a range of H(2)O(2) concentrations. Induction of cell death, proliferation, and migration were investigated using morphology, molecular biology, and biochemical techniques. RESULTS: Human HSC/MFs were shown to be extremely resistant to induction of cell death by O(2)(.-) and only high rates of O(2)(.-) generation induced either necrotic or apoptotic cell death. Non cytotoxic low levels of O(2)(.-), able to upregulate procollagen type I expression (but not tissue inhibitor of metalloproteinase 1 and 2), stimulated migration of human HSC/MFs in a Ras/extracellular regulated kinase (ERK) dependent, antioxidant sensitive way, without affecting basal or platelet derived growth factor (PDGF) stimulated cell proliferation. Non-cytotoxic levels of H(2)O(2) did not affect Ras/ERK or proliferative response. A high rate of O(2)(. ) generation or elevated levels of H(2)O(2 )induced cytoskeletal alterations, block in motility, and inhibition of PDGF dependent DNA synthesis. CONCLUSIONS: Low non-cytotoxic levels of extracellularly generated O(2)(.-) may stimulate selected profibrogenic responses in human HSC/MFs without affecting proliferation. PMID- 16041065 TI - Structure determination of a novel protein by sulfur SAD using chromium radiation in combination with a new crystal-mounting method. AB - A novel and easy crystal-mounting technique was developed for the sulfur SAD method using Cr Kalpha radiation (2.29 A). Using this technique, the cryo-buffer and cryoloop around the protein crystal can be removed before data collection in order to eliminate their X-ray absorption. The superiority and reproducibility of the data sets with this mounting technique were demonstrated using tetragonal hen egg-white lysozyme crystals. The structure of a novel protein, PH1109, from Pyrococcus horikoshii OT3 was solved using this technique. At the wavelength of Cr Kalpha radiation, the anomalous signal |DeltaF|/|F| of PH1109 is expected to be 1.72% as this protein of 144 residues includes four methionines and two cysteines. Sulfur SAD phasing was performed using SHELXD and SHELXE. In the case of the data set obtained using this novel crystal-mounting technique, 54.9% of all residues were built with side chains automatically by RESOLVE. On the other hand, only 16.0% were built with side chains for the data set collected using the standard cryoloop. These results indicated that this crystal-mounting technique was superior to the standard loop-mounting method for the measurement of small anomalous differences at longer wavelength and yielded better results in sulfur substructure solution and initial phasing. The present study demonstrates that the sulfur SAD method with a chromium source becomes enhanced and more practical for macromolecular structure determination using the new crystal-mounting technique. PMID- 16041066 TI - Getting the most out of X-ray home sources. AB - The structures of a 14 kDa phospholipase, an 18 kDa proteinase inhibitor and a novel glycoside hydrolase with molecular weight 60 kDa were solved using the SAD technique and the effects of the amount of anomalous signal, completeness and redundancy of data on heavy-atom substructure determination, phasing and model building were analyzed. All diffraction data sets were collected on a Cu-anode X ray home source. The structure of the phospholipase was obtained using the anomalous scattering contribution from its 16 S atoms. Three-dimensional models for the other two macromolecules were obtained using the anomalous contribution of I atoms rapidly incorporated into the crystal through the quick cryo-soaking method of derivatization. These results were used to discuss the application of sulfur- and iodine-SAD approaches in combination with X-ray home sources for high throughput protein crystal structure solution. The estimates of the anomalous signal from S atoms in the gene products of four genomes are given and the prospects for increasing the anomalous contribution using longer wavelengths (e.g. from a chromium home source) and quick cryo-soaking derivatization are discussed. The possibility of rapidly preparing tangible home-source isomorphous derivatives suggests that this approach might become a valuable tool in the future of post-genomic projects. PMID- 16041067 TI - High-resolution structure of HLA-A*1101 in complex with SARS nucleocapsid peptide. AB - The structure of the human MHC-I molecule HLA-A*1101 in complex with a nonameric peptide (KTFPPTEPK) has been determined by X-ray crystallography to 1.45 A resolution. The peptide is derived from the SARS-CoV nucleocapsid protein positions 362-370 (SNP362-370). It is conserved in all known isolates of SARS-CoV and has been verified by in vitro peptide-binding studies to be a good to intermediate binder to HLA-A*0301 and HLA-A*1101, with IC50 values of 70 and 186 nM, respectively [Sylvester-Hvid et al. (2004), Tissue Antigens, 63, 395-400]. In terms of the residues lining the peptide-binding groove, the HLA-A*1101-SNP362 370 complex is very similar to other known structures of HLA-A*1101 and HLA A*6801. The SNP362-370 peptide is held in place by 17 hydrogen bonds to the alpha chain residues and by nine water molecules which are also tightly bound in the peptide-binding groove. Thr6 of the peptide (Thr6p) does not make efficient use of the middle (E) pocket. For vaccine development, there seems to be a potential for optimization targeted at this position. All residues except Thr2p and Lys9p are accessible for T-cell recognition. PMID- 16041068 TI - A modified vapor-diffusion crystallization protocol that uses a common dehydrating agent. AB - In the vapor-diffusion protein-crystallization method, a small drop containing protein sample mixed with a crystallization solution is equilibrated against a reservoir solution in a sealed chamber. Whereas the chemical composition of the crystallization solution is critical for success, the primary role of the reservoir solution is to slowly concentrate the crystallization drop in a controlled fashion. Accordingly, it might be possible to use any reservoir solution of appropriate dehydrating strength. The important practical consequence is that many different experiments can share the same reservoir solution. This approach, called the ;shared reservoir solution' method, significantly simplifies manual and robotic experiment setup, reduces cost and allows a completely new design of optically superior and higher density crystallization plates. Although this research was motivated by these practical advantages, recent reports and the authors' results indicate that this method may actually increase crystallization success. The authors suggest that this may indicate that a protein has a preferred water activity for crystallization. Here, present practical and theoretical considerations as well as experimental tests of the shared reservoir solution method are presented. PMID- 16041069 TI - Structure of the uracil-DNA N-glycosylase (UNG) from Deinococcus radiodurans. AB - Uracil-DNA glycosylases are DNA-repair enzymes that catalyse the removal of promutagenic uracil from single- and double-stranded DNA, thereby initiating the base-excision repair (BER) pathway. Uracil in DNA can occur by mis-incorporation of dUMP in place of dTMP during DNA synthesis or by deamination of cytosine, resulting in U-A or U-G mispairs. The radiation-resistant bacterium Deinococcus radiodurans has an elevated number of uracil-DNA glycosylases compared with most other organisms. The crystal structure of dr0689 (uracil-DNA N-glycosylase), which has been shown to be the major contributor to the removal of mis incorporated uracil bases in crude cell extracts of D. radiodurans, is reported. PMID- 16041070 TI - De novo calcium/sulfur SAD phasing of the human formylglycine-generating enzyme using in-house data. AB - Sulfatases are a family of enzymes essential for the degradation of sulfate esters. Formylglycine is the key catalytic residue in the active site of sulfatases and is generated from a cysteine residue by FGE, the formylglycine generating enzyme. Inactivity of FGE owing to inherited mutations in the FGE gene results in multiple sulfatase deficiency (MSD), which leads to early death in infants. Human FGE was crystallized in the presence of traces of the protease elastase, which was absolutely essential for crystal growth, and the structure of FGE was determined by molecular replacement. Before this model was completed, the FGE structure was re-determined by SAD phasing using in-house data based on the anomalous signal of two calcium ions bound to the native enzyme and intrinsic S atoms. A 14-atom substructure was determined at 1.8 A resolution by SHELXD; SHELXE was used for density modification and phase extension to 1.54 A resolution. Automated model building with ARP/wARP and refinement with REFMAC5 yielded a virtually complete model without manual intervention. The minimal data requirements for successful phasing and the relative contributions of the Ca and S atoms are discussed and compared with the related FGE paralogue, pFGE. This work emphasizes the usefulness of de novo phasing using weak anomalous scatterers and in-house data. PMID- 16041071 TI - Structure of myelin P2 protein from equine spinal cord. AB - Equine P2 protein has been isolated from horse spinal cord and its structure determined to 2.1 A. Since equine myelin is a viable alternative to bovine tissue for large-scale preparations, characterization of the proteins from equine spinal cord myelin has been initiated. There is an unusually high amount of P2 protein in equine CNS myelin compared with other species. The structure was determined by molecular replacement and subsequently refined to an R value of 0.187 (Rfree=0.233). The structure contains a molecule of the detergent LDAO and HEPES buffer in the binding cavity and is otherwise analogous to other cellular retinol binding proteins. PMID- 16041072 TI - Structure of the wild-type TEM-1 beta-lactamase at 1.55 A and the mutant enzyme Ser70Ala at 2.1 A suggest the mode of noncovalent catalysis for the mutant enzyme. AB - One of the best-studied examples of a class A beta-lactamase is Escherichia coli TEM-1 beta-lactamase. In this class of enzymes, the active-site serine residue takes on the role of a nucleophile and carries out beta-lactam hydrolysis. Here, the structures of the wild-type and the S70G enzyme determined to 1.55 and 2.1 A, respectively, are presented. In contrast to the previously reported 1.8 A structure, the active site of the wild-type enzyme (1.55 A) structure does not contain sulfate and Ser70 appears to be in the deprotonated form. The X-ray crystal structure of the S70G mutant has an altered Ser130 side-chain conformation that influences the positions of water molecules in the active site. This change allows an additional water molecule to be positioned similarly to the serine hydroxyl in the wild-type enzyme. The structure of the mutant enzyme suggests that this water molecule can assume the role of an active-site nucleophile and carry out noncovalent catalysis. The drop in activity in the mutant enzyme is comparable to the drop observed in an analogous mutation of the nucleophilic serine in alkaline phosphatase, suggesting common chemical principles in the utilization of nucleophilic serine in the active site of different enzymes. PMID- 16041073 TI - Ab initio phasing at resolution higher than experimental resolution. AB - Owing to the limited experimental resolution of data in macromolecular crystallography, ab initio phasing is successful only when atomic or quasi-atomic resolution data are available. It is shown that extrapolating the moduli and phases of non-measured reflections beyond and behind the experimental resolution limit makes the ab initio phasing process more efficient and leads to crystal structure solution even in cases in which the standard SIR2004 program does not succeed. Moreover, use of the extrapolated values improves the quality of the final electron-density maps and makes the recognition of the correct structure among several trial structures easier. PMID- 16041074 TI - The hydration structure of a Z-DNA hexameric duplex determined by a neutron diffraction technique. AB - In order to reveal the hydration structure of Z-DNA, a neutron diffraction study has been carried out at 1.8 A resolution on a Z-DNA hexamer d(CGCGCG). Neutron diffraction data were collected with the BIX-3 single-crystal diffractometer at the JRR-3 reactor in the Japan Atomic Energy Research Institute (JAERI) using a large crystal (1.6 mm3) obtained from D2O solution. It has been found that almost all the guanine bases have participated in H/D exchange at the C8-H8 group, consistent with the acidic nature of this bond. 44 water molecules were found in the nuclear density maps, of which 29 showed the entire contour of all three atoms (D-O-D). The remaining 15 water molecules had a simple spherical shape, indicating that they were rotationally disordered. An interesting relationship was found between the orientational disorder of the water molecules and their locations. Almost all water molecules in the minor groove were well ordered in the crystal, while 40% of the water molecules in the major groove were rotationally disordered. The hydrogen-bonding networks in the hydration shells have two structural aspects: flexibility and regularity. PMID- 16041075 TI - Ab initio crystal structure determination of spherical viruses that exhibit a centrosymmetric location in the unit cell. AB - Ab initio phasing by non-crystallographic symmetry averaging coupled with solvent flattening has previously been used to determine the structure of canine parvovirus. As CPV particles were located at general positions, initial phases were generated with a spherical shell deviating from the centre of symmetry. In many virus crystals, the viral particles are located at positions with a centre of symmetry in the unit cell. Thus, the initial phases calculated with a spherical shell model have a centre of symmetry. The inherent difficulty in structural determination was breaking the centre of symmetry in the initial phase. The centric nature of the initial phases of the rice dwarf virus crystal, however, was successfully broken at low resolution by iteration of the density modification method described by Tsao et al. (1992). In this study, ab initio phasing was tested for seven viruses ranging from 82 to 344 A in radius. Although the crystal structures of the initial spherical shell models for each virus had a centre of symmetry, the centric natures of the initial phases were successfully broken by non-crystallographic symmetry averaging coupled with solvent flattening at a low resolution; these phases were then successfully extended to high resolution measurements. A novel procedure of ab initio phasing for spherical virus crystals is proposed. PMID- 16041076 TI - Structure of the zinc-saturated C-terminal lobe of bovine lactoferrin at 2.0 A resolution. AB - The crystal structure of the zinc-saturated C-terminal lobe of bovine lactoferrin has been determined at 2.0 A resolution using crystals stabilized at pH 3.8. This is the first metal-saturated structure of any functional lactoferrin at such a low pH. Purified samples of proteolytically generated zinc-saturated C-terminal lobe were crystallized from 0.1 M MES buffer pH 6.5 containing 25%(v/v) polyethyleneglycol monomethyl ether 550 and 0.1 M zinc sulfate heptahydrate. The crystals were transferred to 25 mM ammonium acetate buffer containing 25%(v/v) polyethyleneglycol monomethyl ether 550 and the pH was gradually changed from 6.5 to 3.8. The X-ray intensity data were collected with a 345 mm imaging-plate scanner mounted on an RU-300 rotating-anode X-ray generator using crystals soaked in the buffer at pH 3.8. The structure was determined with the molecular replacement method using the coordinates of the monoferric C-terminal lobe of bovine lactoferrin as a search model and was refined to an R factor of 0.192 for all data to 2.0 A resolution. The final model comprises 2593 protein atoms (residues 342-676 and 681-685), 138 carbohydrate atoms (from 11 monosaccharide units in three glycan chains), three Zn2+ ions, one CO3(2-) ion, one SO(4)2- ions and 227 water molecules. The overall folding of the present structure is essentially similar to that of the monoferric C-terminal lobe of bovine lactoferrin, although it contains Zn2+ in place of Fe3+ in the metal-binding cleft as well as two additional Zn2+ ions on the surface of the C-terminal lobe. The Zn2+ ion in the cleft remains bound to the lobe with octahedral coordination. The bidentate carbonate ion is stabilized by a network of hydrogen bonds to Ala465, Gly466, Thr459 and Arg463. The other two zinc ions also form sixfold coordinations involving symmetry-related protein and water molecules. The number of monosaccharide residues from the three glycan chains of the C-terminal lobe was 11, which is the largest number observed to date. The structure shows that the C-terminal lobe of lactoferrin is capable of sequestering a Zn2+ ion at a pH of 3.8. This implies that the zinc ions can be sequestered over a wide pH range. The glycan chain attached to Asn545 may also have some influence on iron release from the C-terminal lobe. PMID- 16041077 TI - The crystal structures of native and (S)-lysine-bound dihydrodipicolinate synthase from Escherichia coli with improved resolution show new features of biological significance. AB - Dihydrodipicolinate synthase (DHDPS) mediates the key first reaction common to the biosynthesis of (S)-lysine and meso-diaminopimelate. The activity of DHDPS is allosterically regulated by the feedback inhibitor (S)-lysine. The crystal structure of DHDPS from Escherichia coli has previously been published, but to only a resolution of 2.5 A, and the structure of the lysine-bound adduct was known to only 2.94 A resolution. Here, the crystal structures of native and (S) lysine-bound dihydrodipicolinate synthase from E. coli are presented to 1.9 and 2.0 A, respectively, resolutions that allow, in particular, more accurate definition of the protein structure. The general architecture of the active site is found to be consistent with previously determined structures, but with some important differences. Arg138, which is situated at the entrance of the active site and is thought to be involved in substrate binding, has an altered conformation and is connected via a water molecule to Tyr133 of the active-site catalytic triad. This suggests a hitherto unknown function for Arg138 in the DHDPS mechanism. Additionally, a re-evaluation of the dimer-dimer interface reveals a more extensive network of interactions than first thought. Of particular interest is the higher resolution structure of DHDPS with (S)-lysine bound at the allosteric site, which is remote to the active site, although connected to it by a chain of conserved water molecules. (S)-Lysine has a slightly altered conformation from that originally determined and does not appear to alter the DHDPS structure as others have reported. PMID- 16041078 TI - Structure of d(TGCGCG).d(CGCGCA) in two crystal forms: effect of sequence and crystal packing in Z-DNA. AB - The sequence d(TGCGCG).d(CGCGCA) crystallized in two crystal forms, orthorhombic and hexagonal, in the presence of cobalt hexammine chloride, a known inducer of the left-handed Z-form of DNA. The crystal structures have been solved and refined at 1.71 A resolution in space group P2(1)2(1)2(1) and 2.0 A resolution in space group P6(5). The orthorhombic structure contains one Z-DNA hexamer duplex, while the hexagonal structure contains two hexamer duplexes in the structure. Of the latter, one is situated on a crystallographic sixfold screw axis, leading to disorder. This paper reports the effects of sequence and crystal packing on the structure of Z-type DNA. The structures lend additional support to the authors' earlier conclusion that a stretch of four C.G base pairs is sufficient to nucleate and define the regular model of the left-handed helix based on the structure of d(CGCGCG)2. PMID- 16041079 TI - Extraction of functional motion in trypsin crystal structures. AB - The analysis of anisotropic atomic displacement parameters for the direct extraction of functionally relevant motion from X-ray crystal structures of Fusarium oxysporum trypsin is presented. Several atomic resolution structures complexed with inhibitors or substrates and determined at different pH values and temperatures were investigated. The analysis revealed a breathing-like molecular motion conserved across trypsin structures from two organisms and three different crystal forms. Directional motion was observed suggesting a change of the width of the substrate-binding cleft and a change in the length of the specificity pocket. The differences in direction of motion across the structures are dependent on the mode of substrate or inhibitor binding and the chemical environment around the active-site residues. Together with the occurrence of multiple-residue conformers, they reflect spatial rearrangement throughout the deacylation pathway. PMID- 16041080 TI - Structure of Mycobacterium smegmatis single-stranded DNA-binding protein and a comparative study involving homologus SSBs: biological implications of structural plasticity and variability in quaternary association. AB - The structure of Mycobacterium smegmatis single-stranded DNA-binding protein (SSB) has been determined using three data sets collected from related crystals. The structure is similar to that of its homologue from Mycobacterium tuberculosis, indicating that the clamp arrangement that stabilizes the dimer and the ellipsoidal shape of the tetramer are characteristic features of mycobacterial SSBs. The central OB fold is conserved in mycobacterial SSBs as well as those from Escherichia coli, Deinococcus radiodurans and human mitochondria. However, the quaternary structure exhibits considerable variability. The observed plasticity of the subunit is related to this variability. The crystal structures and modelling provide a rationale for the variability. The strand involved in the clamp mechanism, which leads to higher stability of the tetramer, appears to occur in all high-G+C Gram-positive bacteria. The higher stability is perhaps required by these organisms. The mode of DNA binding of mycobacterial SSBs is different from that of E. coli SSB partly on account of the difference in the shape of the tetramers. Another difference between the two modes is that the former contains additional ionic interactions and is more susceptible to salt concentration. PMID- 16041081 TI - Impact of natural variation in bacterial F17G adhesins on crystallization behaviour. AB - Since the introduction of structural genomics, the protein has been recognized as the most important variable in crystallization. Recent strategies to modify a protein to improve crystal quality have included rationally engineered point mutations, truncations, deletions and fusions. Five naturally occurring variants, differing in 1-18 amino acids, of the 177-residue lectin domain of the F17G fimbrial adhesin were expressed and purified in identical ways. For four out of the five variants crystals were obtained, mostly in non-isomorphous space groups, with diffraction limits ranging between 2.4 and 1.1 A resolution. A comparative analysis of the crystal-packing contacts revealed that the variable amino acids are often involved in lattice contacts and a single amino-acid substitution can suffice to radically change crystal packing. A statistical approach proved reliable to estimate the compatibilities of the variant sequences with the observed crystal forms. In conclusion, natural variation, universally present within prokaryotic species, is a valuable genetic resource that can be favourably employed to enhance the crystallization success rate with considerably less effort than other strategies. PMID- 16041082 TI - Structure of the lipopeptide antibiotic tsushimycin. AB - The amphomycin derivative tsushimycin has been crystallized and its structure determined at 1.0 A resolution. The asymmetric unit contains 12 molecules and with 1300 independent atoms this structure is one of the largest solved using ab initio direct methods. The antibiotic is comprised of a cyclodecapeptide core, an exocyclic amino acid and a fatty-acid residue. Its backbone adopts a saddle-like conformation that is stabilized by a Ca2+ ion bound within the peptide ring and accounts for the Ca2+-dependence of this antibiotic class. Additional Ca2+ ions link the antibiotic molecules to dimers that enclose an empty space resembling a binding cleft. The dimers possess a large hydrophobic surface capable of interacting with the bacterial cell membrane. The antibiotic daptomycin may exhibit a similar conformation, as the amino-acid sequence is conserved at positions involved in Ca2+ binding. PMID- 16041083 TI - A nanovolume crystallization robot that creates its crystallization screens on the-fly. AB - Protein crystallization generally consists of an initial screen followed by optimization of promising conditions. Whereas the initial screen typically uses a standard set of pre-made crystallization cocktails, optimization requires new cocktails with small perturbations of the original composition. Highly parallel synchronous crystallization robots are ideal for initial screening, but they depend on pre-made crystallization cocktails. Asynchronous crystallization robots can create crystallization cocktails from stock solutions, but in practice this ability is rarely exploited. Instead, large-scale operations typically use a general liquid-handling robot to create optimization screens, whereas academics mostly rely on manual optimization. Here, the use of an asynchronous crystallization robot to create customized crystallization cocktails and set up nanovolume crystallization experiments without a compromise in speed or drop quality is described. This approach avoids the complex integration of hardware, software and dataflow between two robots and saves cost and space. As a proof of principle, a commercial crystal screen has been reproduced with the robot and shows that results are virtually identical to using the actual commercial screen. PMID- 16041085 TI - Structure of KNbOB2O5--a commensurately modulated structure. AB - Members of the AMOB2O5 (A = K, Rb, Cs, Tl; M = Nb, Ta) family of compounds can be described as modulated structures with a single superspace group and very similar modulation functions. Single-crystal X-ray diffraction data (Mo Kalpha radiation) are used to solve and refine the structure of KNbOB2O5 in these terms for the first time. The average structure is solved and refined in the space group Pmn2(1). Subsequently, the atomic modulation functions are determined using JANA2000 and superspace-group symmetry Pmn2(1)(0,0.375,0)s. The commensurately modulated structure is finally refined as a superstructure in the space group Pbn2(1) using SHELXS97 converging to R(1) = 0.024. PMID- 16041086 TI - Structure of the clinopyroxene-type compound CaCuGe2O6 between 15 and 800 K. AB - CaCuGe2O6 shows a strongly distorted clinopyroxene-type structure with P2(1)/c symmetry at 298 K. The Cu2+ ion at the M1 site is coordinated by six O atoms forming an octahedron, which deviates significantly from ideal geometry. Individual M1 sites are connected via common edges to form an infinite zigzag chain parallel to the crystallographic c axis. The Ca2+ ion at M2 shows a sevenfold coordination. M2 sites are connected to the M1 chain via three common edges, thereby forming a metal layer within the bc plane. Besides the strong Jahn Teller distortion of the Cu site, the structure of the title compound differs from ;normal' clinopyroxenes by a distortion of alternate layers of Ge sites. While the Ge(A) site is fourfold coordinated by O atoms, forming infinite chains of corner-sharing chains parallel to the c axis, the Ge(B) site exhibits a fivefold coordination, thereby forming a true two-dimensional layer of edge sharing GeO5 bipyramids. Decreasing the temperature causes a magnetic phase transition at 40 K, as monitored by a broad maximum in the magnetic susceptibility and by discontinuities in the lattice parameters. Increasing the temperature causes variations in bond lengths, edge lengths and bond angles. Most prominent is the increase of one bond length of the Ge(B) site and the increase of the tetrahedral bridging angle of the Ge(A) site. At 660 K a crystallographic phase transition is observed where the symmetry changes from P2(1)/c to C2/c. The transition is accompanied by large changes in the lattice parameters which are indicative of distinct topological changes of several structural building units. The high-temperature C2/c structure is similar to that of the germanate clinopyroxene CaMgGe2O6. PMID- 16041087 TI - Structure of strontium hydroxide octahydrate, Sr(OH)2.8H2O, at 20, 100 and 200 K from neutron diffraction. AB - The crystal structure of Sr(OH)2*8H2O has been determined at 20, 100 and 200 K from neutron diffraction data. The structure consists of double layers of H2O and OH- ions separated by Sr2+ along the c axis. The Sr2+ ions are eight-coordinated by water O atoms in a square antiprism configuration. Each H2O molecule is engaged in three hydrogen bonds. The OH- ions form chains of acceptor and donor bonds along the fourfold axis with O atoms engaged in four bonds with H2O molecules, such that both non-equivalent O atoms have square-pyramidal environments of five H atoms and the overall bonding configurations of distorted octahedra. PMID- 16041088 TI - Electron diffraction of tilted perovskites. AB - Simulations of electron diffraction patterns for each of the known perovskite tilt systems have been performed. The conditions for the appearance of superlattice reflections arising from rotations of the octahedra are modified to take into account the effects of different tilt systems for kinematical diffraction. The use of selected-area electron diffraction as a tool for perovskite structure determination is reviewed and examples are included. PMID- 16041089 TI - The role of second coordination-sphere interactions in incommensurately modulated structures, using beta-K5Yb(MoO4)4 as an example. AB - The incommensurate palmierite-like structure of beta-K5Yb(MoO4)4, potassium ytterbium tetramolydate, has been refined in the (3 + 1)-dimensional monoclinic superspace group X2/m(0rho0)00, with X = [0 0 0 0; (1/2) (1/2) 0 0; 0 0 (1/2) (1/2); (1/2) (1/2) (1/2) (1/2)] and the unit-cell parameters a = 10.4054 (16), b = 6.1157 (12), c = 19.7751 (18) A, beta = 136.625 (10) degrees ; q = 0.6354 (30)b*. The occupations of the K and Yb atomic positions are described by crenel functions. The structure model reveals a balanced interaction between the atoms of the first and second coordination spheres. It is shown that the third coordination sphere should not be neglected in studies of modulated structures. The ordering of the K and Yb atoms appears to be the driving force for the modulation of all the other atoms. PMID- 16041090 TI - Structure refinement of a twinned pseudo-symmetric crystal of [Mn(C10H24N4)(NCO)2]+*ClO4-. AB - The crystal studied is a 0.545 (1):0.455 twin, space group C?bar 1, Z = 16, and is a commensurate occupational and displacive modulation of a Z = 4 idealized parent structure with the space group A2/a and a(p) = a/2, b(p) = b/2, c(p) = c. A hierarchical approach to solution and refinement led sequentially to structures in the space groups A2/a, P2(1)/n, P?bar 1 and finally C?bar 1. The major and minor components of the reflection intensities could be identified using irreducible representations of A2/a and P2(1)/n, which in turn suggested suitable constraints and restraints for optimizing the refinement pathway. Comparative refinement was used to show the correctness of the final structure solution and how appropriately chosen constrained refinement allowed an escape from a false minima. PMID- 16041091 TI - Atomic interactions in ethylenebis(1-indenyl)zirconium dichloride as derived by experimental electron density analysis. AB - A topological analysis of the experimental electron density in racemic ethylenebis(1-indenyl)zirconium dichloride, C20H16Cl2Zr, measured at 100 (1) K, has been performed. The atomic charges calculated by the numerical integration of the electron density over the zero-flux atomic basins demonstrate the charge transfer of 2.25 e from the Zr atom to the two indenyl ligands (0.19 e to each) and two Cl atoms (0.93 e to each). All the atomic interactions were quantitatively characterized in terms of the electron density and the electronic energy-density features at the bond critical points. The Zr-C2 bond paths significantly curved towards the C1-C2 bond were found; no other bond paths connecting the Zr atom and indenyl ligand were located. At the same time, the pi electrons of the C1-C2 bond are significantly involved in the metal-ligand interaction. The electron density features indicate that the indenyl coordination can be approximately described as eta1 with slippage towards eta2. The ;ligand opposed' charge concentrations around the Zr atom were revealed using the Laplacian of the electron density and the one-particle potential; they were linked to the orbital representations. Bonds in the indenyl ligand were characterized using the Cioslowski-Mixon bond-order indices calculated directly from the experimental electron density. PMID- 16041092 TI - New lanthanide-nitrogen bond-valence parameters. AB - The bond-valence parameters (R(ij)), which connect bond valences and bond lengths, have been computed for lanthanide-nitrogen bonds. It has been found that values of bond-valence parameters decrease with increasing lanthanide atomic number in coordination compounds, and that they are smaller than the R(ij) parameters of inorganic compounds. As expected, the lanthanide-nitrogen bond valence parameters are larger than lanthanide-oxygen bond-valence parameters. There are no obvious dependencies between the number of N atoms in the coordination sphere and the bond-valence parameter value. PMID- 16041093 TI - Powder diffraction crystal structure analysis using derivative difference minimization: example of the potassium salt of 1-(tetrazol-5-yl)-2 nitroguanidine. AB - The crystal structure of the potassium salt of 1-(tetrazol-5-yl)-2-nitroguanidine [K(C2H3N8O2)] was solved and refined from X-ray powder diffraction data by applying the derivative difference minimization (DDM) method. The compound is of interest as an energetic substance. The structure model was found from a Patterson search. The reflection intensities for the Patterson synthesis were derived from the powder profile by applying a newly developed DDM-based profile decomposition procedure. The use of the DDM method allowed successful location and unconstrained refinement of all the atomic positions, including those of three independent H atoms. The advantages of DDM in terms of the precision and reproducibility of the structural parameters are discussed in comparison to Rietveld refinement results. The failure to refine the H-atom positions by the Rietveld method was attributed to systematic errors associated with the background modelling, which are avoided by DDM. PMID- 16041094 TI - Submolecular partitioning of morphine hydrate based on its experimental charge density at 25 K. AB - The electron density distribution of morphine hydrate has been determined from high-resolution single-crystal X-ray diffraction measurements at 25 K. A topological analysis was applied and, in order to analyze the submolecular transferability based on an experimental electron density, a partitioning of the molecule into atomic regions was carried out, making use of Bader's zero-flux surfaces to yield atomic volumes and charges. The properties obtained were compared with the theoretical calculations of smaller fragment molecules, from which the complete morphine molecule can be reconstructed, and with theoretical studies of another opiate, Oripavine PEO, reported in the literature. PMID- 16041095 TI - The low-temperature and high-pressure crystal structures of cyclobutanol (C4H7OH). AB - The low-temperature and high-pressure crystal structures of cyclobutanol (C4H7OH) have been determined using single-crystal X-ray diffraction techniques. At temperatures below 220 K, cyclobutanol crystallizes in the Aba2 space group (Z' = 2) and its crystal structure is composed of pseudo-threefold hydrogen-bonded molecular catemers [assigned as C_2;2(4) in graph-set notation], which lie parallel to the crystallographic a axis. At a pressure of 1.3 GPa, the crystal symmetry changes to Pna2(1) (Z' = 1) and the molecular catemers [expressed as C2 in graph-set notation] adopt a pseudo-twofold arrangement. This structural behaviour is in agreement with our previous observations for phenol and its halogenated derivatives 2-chlorophenol and 4-fluorophenol, where pressure was found to favour a molecular packing more closely associated with small alkyl groups rather than that of relatively bulky alkyl groups. In addition, an examination of the molecular coordination environment in the low-temperature and high-pressure structures of cyclobutanol reveals that the change in structure on application of pressure appears to be driven by the molecules assuming a packing arrangement which more closely resembles that adopted in hard-sphere structures. PMID- 16041096 TI - Structure determination of the 1/1 alpha/beta mixed lactose by X-ray powder diffraction. AB - The mixed form of alpha/beta lactose was obtained by heating amorphous alpha lactose at 443 K. NMR spectroscopy determined the stoichiometry of this mixed compound to be 1/1. The X-ray powder diffraction pattern was recorded at room temperature with a sensitive curved detector (CPS 120). The structure was solved by real-space methods (simulated annealing) followed by Rietveld refinements with soft constraints on bond lengths and bond angles. The H atoms of the hydroxyl groups were localized by minimization of the crystalline energy. The cell of 1/1 alpha/beta lactose is triclinic with the space group P1 and contains two molecules (one molecule of each anomer). The crystalline cohesion is achieved by networks of O-H...O hydrogen bonds. The width of the Bragg peaks is interpreted through a microstructural approach in terms of isotropic strain effects and anisotropic size effects. PMID- 16041097 TI - A variable-temperature study of a phase transition in barbituric acid dihydrate. AB - The crystal structure of barbituric acid dihydrate (C4H4N2O3*2H2O) has twice been reported as orthorhombic, space group Pnma, with all atoms (except for CH(2) H atoms) lying on the mirror plane [Al-Karaghouli et al. (1977). Acta Cryst. B33, 1655-1660; Jeffrey et al. (1961). Acta Cryst. 14, 881-887]. The present study has found that at low temperatures, below 200 K, the crystal structure is no longer orthorhombic but is non-merohedrally twinned monoclinic, space group P2(1)/n. This phase is stable down to 100 K. Above 220 K the crystal structure is orthorhombic, and between 200 and 220 K the structure undergoes a phase change, with the monoclinic-to-orthorhombic phase transition itself taking place at around 216-217 K. The size of the beta angle in the monoclinic structure is temperature dependent; at 100 K beta is around 94 degrees and it decreases in magnitude towards 90 degrees as the temperature increases. Although the hydrogen bonding motifs are the same for both crystal systems, there are significant differences in the crystal packing, in particular the out-of-plane displacement of the two water molecules and the sp3-hybridized C atom of barbituric acid. PMID- 16041098 TI - Training in laparoscopy--which model to use? PMID- 16041099 TI - Dietary factors in pathogenesis of gallstone disease in southern India--a hospital-based case-control study. AB - BACKGROUND: Pigment or mixed gallstones are common in southern India. The etiology is not established. Known risk factors include an obese, diabetic female and a nonsmoker male. AIM: To determine the association of dietary factors with mixed/pigment gallstones amongst southern Indian patients. METHODS: Diet details were obtained from 346 patients (178 women) with gallstones and an equal number of healthy controls who were age- and sex-matched attendants of the patients, sharing similar socioeconomic and demographic characteristics, with normal abdominal ultrasonogram. Data recorded included the number of daily meals, nature of cereal used, vegetarianism, oil consumed per month, sugar consumption per day, tamarind (Garcinia camborginia ) usage per week, and per-day beverage consumption. RESULTS: There was no difference between cases and controls in consumption of non-vegetarian food, type of cereal, average oil and sugar consumption, and type of beverage consumed (tea/coffee/milk/combination). Individuals with BMI> 22 were at higher risk to develop gallstones (OR 1.49; 95% CI 1.09, 2.04; p=0.01). There was significant risk of gallstone formation with the use of tamarind when consumed > 3 times a week (OR 1.76; 95% CI 1.05, 2.96; p=0.03). Higher BMI and tamarind use were significant risk factors even on multivariate logistic regression analysis (p=0.02). CONCLUSION: Higher BMI and use of tamarind, a common ingredient of diet in southern India, are risk factors in the formation of gallstones in southern India. PMID- 16041100 TI - PD98059 inhibits expression of pERK1 protein and collagen alpha1(I) mRNA in rat pancreatic stellate cells activated by platelet-derived growth factor. AB - OBJECTIVE: Pancreatic stellate cells (PSC) are considered as the principal effector cells in pancreatic fibrosis. We studied the role of platelet-derived growth factor (PDGF) in the activation of PSC. METHODS: Cultured rat PSC were co incubated with PDGF-BB (25 ng/mL) and different doses (0-40 ng/mL) of PD98059, a specific inhibitor of extracellular signal-regulated kinase (ERK). Expressions of p ERK1 protein and of collagen a1(I) mRNA were measured. RESULTS: Expression of p ERK1 protein was up-regulated by PDGF-BB, and was down-regulated in a dose dependent manner by PD98059. Expression of collagen a1(I) mRNA also showed an increase with PDGF-BB and non-dose-dependent inhibition by PD98059. CONCLUSION: Our findings suggest that PSC activation is mediated by PDGF signal pathway, and ERK1 protein plays an important role in this activation. PMID- 16041101 TI - Early stereological changes in liver of Sprague-Dawley rats after streptozotocin injection. AB - BACKGROUND: Diabetes mellitus is associated with biochemical, physiological and pathologic alterations in the liver. We measured changes in structure of rat liver after streptozotocin injection, using stereology. METHODS: Livers of 36 streptozotocin-injected rats were removed after 4, 8 and 12 weeks. Liver volume and weight were measured, and volume-weighted mean volume of hepatocytes and their nuclei were estimated in periportal (Z1), interstitial (Z2) and perivenous (Z3) zones of liver acini. Volume of liver sinusoids was also estimated. RESULTS: Mean volume and weight of the liver were reduced by 15% and 12%, respectively at 4 and 8 weeks after injection. Mean hepatocyte volumes were reduced by approximately 30%, 31% and 24% in Z1, Z2 and Z3 at 4 weeks, 19% and 24% in Z2 and Z3 at 8 weeks, and 14% in Z1 at 12 weeks. Mean volume of hepatocyte nuclei was reduced by approximately 18% and 20% in Z2 and Z3 at 4 weeks, 23% in all three zones at 8 weeks, and 18%, 15% and 13% in Z1, Z2 and Z3, respectively, at 12 weeks. The absolute volume of the sinusoids decreased by 16.5% only at 4 weeks. CONCLUSION: Streptozotocin injection leads to early reduction in volume of hepatocytes, their nuclei and sinusoids in rat liver. PMID- 16041102 TI - Role of overnight rifampin test in diagnosing Gilbert's syndrome. AB - BACKGROUND: Gilbert's syndrome (GS) is the most common inherited disorder of bilirubin metabolism. Recent data show that the rifampin test can be used as a diagnostic test but there is controversy about its effect on bilirubin level in normal individuals. We studied the effect of administration of rifampin on serum bilirubin level in patients with GS and in healthy individuals. METHODS: Serum total and unconjugated bilirubin levels were measured in 16 patients with GS and 15 healthy individuals before and after a single 600-mg oral dose of rifampin. RESULTS: In patients with GS, mean (SD) serum total and unconjugated bilirubin level increased from 2.15 (0.49) and 1.56 (0.41) mg/dL, respectively to 3.23 (0.72) (p< 0.001) and 2.52 (0.71) mg/dL (p< 0.001), respectively after rifampin administration, and in healthy subjects from 0.69 (0.13) and 0.34 (0.09) mg/dL, respectively to 1.68 (0.56) (p< 0.001) and 0.84 (0.23) mg/dL (p< 0.001), respectively. Elevation of these levels above the normal cut-off levels had poor accuracy for the diagnosis of GS. However, elevation of total serum bilirubin after rifampin above 2.4 mg/dL was 93.8% sensitive and 93.3% specific for the diagnosis of GS, and elevation of unconjugated bilirubin above 1.3 mg/dL was 100% sensitive and 100% specific. CONCLUSIONS: Rifampin elevates bilirubin level to above normal in GS and healthy subjects. Overnight rifampin test may be useful for the diagnosis of GS if cut-off levels for serum total and unconjugated bilirubin level of more than 2.4 and 1.3 mg/dL are used. PMID- 16041103 TI - Cadaver as a model for laparoscopic training. AB - BACKGROUND: Though minimally invasive techniques now are routine world over, there is need to develop facilities for training surgeons. Laparoscopy performed on anesthetized animals is an established model but is costly and is not easily available. We report on human cadaver as a training modality for surgeons participating in a laparoscopic training course. METHODS: Unembalmed cadavers were used for training surgeons to appreciate anatomy, practice laparoscopic techniques, and deploy equipment and instruments during a laparoscopic training course. Trainees carried out procedures such as cholecystectomy, appendicectomy, splenectomy, intestinal explorations, mesenteric lymph node biopsy, and varicocele-vein occlusion. We analyzed the trainees' perspective regarding cadaver as a model using the 5-point Likert scale. RESULTS: Thirty-two trainees from five consecutive training courses held at our institution expressed general satisfaction over cadaver as a training model, and 96.9% (31/32) rated the training model as highly satisfactory. The trainees ranked as highly satisfactory their understanding of surgical anatomy (29/32; 90.6%), understanding of laparoscopic technique (29/32; 90.6%) and use of instruments (32/32; 100%). The trainees thought such an approach improved spatial perception of anatomy and they perceived it as a valuable educational experience. CONCLUSIONS: Human cadaveric laparoscopy may offer an ideal surgical environment for laparoscopy training courses, allowing dissection and performance of complicated procedures. PMID- 16041104 TI - Band ligation of Dieulafoy lesions. AB - Various endoscopic methods have been used successfully to treat Dieulafoy lesions. We describe our experience at one center of successful band ligation of Dieulafoy lesions in 7 patients. PMID- 16041105 TI - Small bowel metastases from esophageal and oropharyngeal cancers. AB - Intestinal metastases are uncommon. Lungs and skin are the common sites of primary tumor. We report two men (52- and 67-year-old) with small bowel metastases from esophageal and oropharyngeal carcinomas. PMID- 16041106 TI - Mixed exocrine-endocrine pancreatic carcinoma in childhood. AB - A 7-year-old boy with mixed exocrine-endocrine pancreatic cancer is presented. This may be the second reported case of such a tumor in childhood. PMID- 16041107 TI - Acute liver failure and severe hemophagocytosis secondary to parvovirus B19 infection. AB - Parvovirus infection presenting as severe hemophagocytosis is extremely rare. We report a 13-year-old girl with acute parvovirus infection who had severe hemophagocytosis resulting in severe pancytopenia and hepatic failure. PMID- 16041108 TI - Primary esophageal T-cell non-Hodgkin's lymphoma. AB - Primary non-Hodgkin's lymphoma of the esophagus is a rare disease. We report a 52 year-old man who had a polypoid mass in the esophagus at endoscopy. Histology was suggestive of non-Hodgkin's lymphoma; immunohistochemistry was positive for CD3, CD45 RO, LCA. He was treated with 6 cycles of CHOP and is disease-free 14 months later. PMID- 16041109 TI - Cobalamine deficiency associated with neuropathy and oral mucosal melanosis in untreated gluten-sensitive enteropathy. AB - A 32-year-old woman was admitted with complaints of difficulty in walking and hypoaesthesia and tingling in her legs. She had short stature and brown-black hyperpigmentation, cheliosis, dental irregularities and scars in the axillary regions. Neurological examination revealed mild, symmetric, predominantly distal weakness of the legs; deep tendon reflexes were depressed. There was glove-and stocking decrease in pinprick and temperature perception but proprioception and light touch were normal. Investigations established a diagnosis of celiac disease; her neurological features improved on gluten-free diet, but oral pigmentation persisted. PMID- 16041110 TI - Herniation of colon following transhiatal esophagectomy. AB - We report a 38-year-old man with intestinal obstruction following transhiatal esophagectomy for carcinoma esophagus; it occurred secondary to herniation of the transverse colon through the esophageal hiatus into the mediastinum. The patient is asymptomatic after reduction of the hernia and repair of the disphragmatic hernia. PMID- 16041111 TI - Congenital segmental dilatation of colon with colonic atresia. AB - Congenital segmental dilatation of the colon belongs to a group of Hirschsprung's like diseases with normal ganglion cells. The presentation is with chronic constipation affecting older children. We report a neonate with congenital segmental dilatation of the colon associated with sigmoid atresia. The child is well after a colostomy. PMID- 16041112 TI - Enteral feeding by fistuloclysis in a midjejunal fistula. AB - Enterocutaneous fistulas are potentially life-threatening complications of gastrointestinal surgery. Nutritional support is the mainstay of management. We report a 32-year-old man who developed an enterocutaneous fistula following surgery for ulcerative colitis. Enteral feeding was attempted by introducing a Foley's catheter through the midjejunal fistula. PMID- 16041113 TI - Effect of Teucrium polium on histology and histochemistry in rat stomach. PMID- 16041114 TI - Prevalence of genotypes G1-G4 of human rotavirus in a hospital setting in New Delhi. PMID- 16041115 TI - Acute viral hepatitis in third trimester of pregnancy. PMID- 16041116 TI - Is sentinel node mapping useful in colorectal carcinoma? PMID- 16041117 TI - Severe esophageal stricture probably resulting from chronic detergent exposure. PMID- 16041118 TI - Helicobacter pylori infection rates in duodenal ulcer patients in a population with high prevalence of infection. PMID- 16041119 TI - A report of post-ERCP Pseudomonas aeruginosa infection outbreak. PMID- 16041120 TI - Cutaneous metastases in carcinoma stomach. PMID- 16041121 TI - Malignant melanoma metastatic to the stomach and duodenum. PMID- 16041122 TI - Angiotensin III as well as angiotensin II regulates water flow through aquaporins in a clam worm. AB - Angiotensin III has been reported to exist in various animals and tissues. The physiological role, however, is still unclear except that brain angiotensin III is a central regulator of vasopressin release. In this study, angiotensin III as well as angiotensin II enhanced an increase in body weight of clam worms of Perinereis sp. under a hypo-osmotic condition and suppressed a decrease in body weight under a hyper-osmotic condition. When clam worms were treated with tetrachloroaurate (III) after angiotensin-treatment, these enhancing and suppressive effects of the angiotensins under hypo- and hyper-osmotic conditions were inhibited. In contrast, when clam worms were pretreated with tetrachloroaurate (III) before angiotensin-treatment, these effects of angiotensins were not inhibited. Since tetrachloroaurate (III) is a representative blocker of aquaporins, these results indicate that angiotensin III as well as angiotensin II regulates water flow through aquaporins in clam worms. PMID- 16041123 TI - Isolation and identification of Sphingomonas sp. that yields tert-octylphenol monoethoxylate under aerobic conditions. AB - Topsoil samples were collected from eight golf courses in Yamaguchi Prefecture, Japan, and enrichment cultures were carried out with a basal-salt medium containing 0.2% 4-tert-octylphenol polyethoxylate (OPPEO) as sole carbon source. OPPEO-degrading activity was detected in one of the samples, from which a strain of OPPEO-degrading bacterium was isolated. The isolated bacterium grew on a nutritionally enriched medium (NE medium) containing 0.2% OPPEO as sole carbon source, and accumulated 4-tert-octylphenol diethoxylate (OP2EO) (63%), 4-tert octylphenol triethoxylate (OP3EO) (14%), and 4-tert-octylphenol monoethoxylate (OP1EO) (2%) after 7 d cultivation under aerobic conditions. The addition of clay mineral (vermiculite) to the medium accelerated the degradation of OP2EO (40%) and OP3EO (4%) to OP1EO (23%). This is the first report about bacteria that can degrade OPPEO to OP1EO under aerobic conditions. The strain was identified as Sphingomonas macrogoltabidus, based on the homology of a 16S rDNA sequence. PMID- 16041124 TI - Change in content of sugars and free amino acids in potato tubers under short term storage at low temperature and the effect on acrylamide level after frying. AB - Changes in the sugar and amino acid contents of potato tubers during short-term storage and the effect on the acrylamide level in chips after frying were investigated. The acrylamide content in chips began to increase after 3 days of storage at 2 degrees C in response to the increase of glucose and fructose contents in the tubers. There was strong correlation between the reducing sugar content and acrylamide level, R(2)=0.873 for fructose and R(2)=0.836 for glucose. The sucrose content had less correlation with the acrylamide content because of its decrease after 4 weeks of storage at 2 degrees C, while the reducing sugar in potato tubers and the acrylamide in chips continued to increase. The contents of the four amino acids, i.e., asparatic acid, asparagine, glutamic acid and glutamine, showed no significant correlation with the acrylamide level. These results suggest that the content of reducing sugars in potato tubers determined the degree of acrylamide formation in chips. The chip color, as evaluated by L* (lightness), was correlated well with the acrylamide content. PMID- 16041125 TI - Effects of a high-pressure treatment on the activity and structure of rabbit muscle proteasome. AB - The effects were assessed of high hydrostatic pressure on the activity and structure of rabbit skeletal muscle proteasome. The pressure effects on the activity were measured by the amount of fluorometric products released from synthetic substrates under pressure and from fluorescein isothiocyanate (FITC) labeled casein after releasing the pressure. The effects on the structure were measured by fluorescene spectroscopy under pressure, and by circular dichroism (CD) spectroscopy and surface hydrophobicity after releasing the pressure. The optimal pressure for the hydrolyzing activity of synthetic peptides was 50 MPa. The degradation of FITC-labeled casein increased linearly with increasing pressure applied up to 200 MPa, and then markedly decreased up to at 400 MPa. The changes in the tertiary structure detected by fluorometric measurement were irreversible, whereas the changes in the secondary structure were small compared with those by heat treatment. The pressure-induced activation of proteasome therefore seems to have been due to a little unfolding of the active sites of proteasome. PMID- 16041126 TI - Effects of edd and pgi disruptions on inosine accumulation in Escherichia coli. AB - Using an inosine-producing mutant of Escherichia coli, the contributions of the central carbon metabolism for overproducing inosine were investigated. Sodium gluconate instead of glucose was tested as a carbon source to increase the supply of ribose-5-phosphate through the oxidative pentose phosphate pathway. The edd (6 phosphogluconate dehydrase gene)-disrupted mutant accumulated 2.5 g/l of inosine from 48 g/l of sodium gluconate, compared with 1.4 g/l of inosine in the edd wild strain. The rpe (ribulose phosphate 3-epimerase gene)-disrupted mutant resulted in low cell growth and low inosine production on glucose and on gluconate. The disruption of pgi (glucose-6-phosphate isomerase gene) was effective for increasing the accumulation of inosine from glucose but resulted in low cell growth. The pgi-disrupted mutant accumulated 3.7 g/l of inosine from 40 g/l of glucose when 8 g/l of yeast extract was added to the medium. Furthermore, to improve effective utilization of adenine, the yicP (adenine deaminase gene) disrupted mutant was evaluated. It showed higher inosine accumulation, of 3.7 g/l, than that of 2.8 g/l in the yicP wild strain when 4 g/l of yeast extract was added to the medium. PMID- 16041127 TI - Acidolysis between triolein and short-chain fatty acid by lipase in organic solvents. AB - Ten kinds of lipases were examined as biocatalysts for the incorporation of short chain fatty acids (acetic, propionic, and butyric acids) into triolein in order to produce one kind of reduced-calorie structured lipids. Trans-esterification (acidolysis) was successfully done in n-hexane by several microbial lipases. Among them, lipase from Aspergillus oryzae was used to investigate the effects of incubation time, substrate molar ratio, and water content on acidolysis. Finally, more than 80% of triolein was incorporated by butyric acid (molar ratio of triolein to butyric acid, 1:10) in the dried n-hexane at 52 degrees C for 72 h. More than 90% of the products was monosubstituent, which was esterified with this short chain fatty acid at the 1-position of the glycerol moiety of triolein. These results suggest that A. oryzae lipase would be a powerful biocatalyst for the synthesis of low caloric oil, such as triacylglycerol containing a mixture of long- and short-chain aliphatic acids. PMID- 16041128 TI - Gene cloning of an endoglucanase from the basidiomycete Irpex lacteus and its cDNA expression in Saccharomyces cerevisiae. AB - A gene (cen1) coding for an endoglucanase I (En-1) was isolated from white rot fungus Irpex lacteus strain MC-2. The cen1 ORF was comprised of 399 amino acid residues and interrupted by 14 introns. The deduced amino acid sequence of the cen1 ORF revealed a multi-domain structure composed of a cellulose-binding domain, a Ser-/Thr-rich linker, and a catalytic domain from the N-terminus. It showed a significant similarity to those of other endoglucanases that belong to family 5 of glycosyl hydrolases. cen1 cDNA was inserted into a yeast expression vector, YEpFLAG-1, and introduced into Saccharomyces cerevesiae. The resulting S. cerevisiae transformant secreted a recombinant En-1 that had enzymatic properties similar to the original En-1. A strong synergistic effect for a degradation of Avicel and phosphoric acid swollen cellulose was observed when recombinant En-1 was used together with a major exo-type cellobiohydrolase I of I. lacteus MC-2. PMID- 16041129 TI - Asp578 in LEU4p is one of the key residues for leucine feedback inhibition release in sake yeast. AB - We identified a new mutation, Asp578Tyr, in alpha-isopropylmalate synthase (a LEU4 gene product) that releases leucine feedback inhibition and causes hyperproduction of isoamyl alcohol (i-AmOH) in sake yeast. Spontaneous sake yeast mutants that express resistance to 5,5,5-trifluoro-DL-leucine (TFL) were isolated, and a mutant strain, TFL20, was characterized at the genetic and biochemical levels. An enzyme assay for alpha-isopropylmalate synthase showed that strain TFL20 was released from feedback inhibition by L-leucine. Furthermore, DNA sequencing of the LEU4 gene for a haploid of the mutant TFL20 revealed that aspartic acid in position 578 changes to tyrosine. A comparison of the three-dimensional structures of wild-type LEU4p and mutant LEU4D578Yp by the homology modeling method showed that Asp578 is important for leucine feedback inhibition. We conclude that the mutation from Asp to Tyr in 578 is a novel change causing release from leucine feedback inhibition. PMID- 16041130 TI - Effects of N-glycosylation and inositol on the ER stress response in yeast Saccharomyces cerevisiae. AB - IRE1 and HAC1 are essential for the unfolded protein response in the endoplasmic reticulum (ER). IRE1- and HAC1-disruptants require high concentrations of inositol for its normal growth. The ALG6, ALG8, and ALG10 genes encode the glucosyltransferases necessary for the completion of the synthesis of the lipid linked oligosaccharide used for the asparagine-linked glycosylation of proteins in that order. Here we show that, given a combination of the hac1 defect with a disruption of ALG6, ALG8, and ALG10, no strains grow on inositol-free medium. However, the growth defect of the hac1-alg10 double disrupted was partially, but significantly, suppressed by the addition of inositol to the medium. These results indicate that inositol, according to the numbers of glucose residues in the oligosaccharide, plays an important role in the stress response and quality control of glycoproteins in the ER. PMID- 16041131 TI - Isolation and molecular characterization of the locked-on mutant of Mg2+ sensor PhoQ in Escherichia coli. AB - A Mg(2+) sensor mutant (PhoQD179L(A)) in which D179 of PhoQ was changed into L or A was isolated and characterized in Escherichia coli. PhoQ-PhoP regulon genes, phoPQ, mgtA and mgrB transcriptions were repressed at a high Mg(2+) concentration in WQ3007 (phoQ-defective strain)/pHO119, but not in WQ3007/pHO179L(A). The in vitro autophosphorylation activity of membrane-bound PhoQ was repressed by Mg(2+) (10 mM), but that of membrane-bound PhoQD179L(A) was not. Furthermore, the phosphotransfer from membrane-bound PhoQ to PhoP was also repressed by Mg(2+), but was not observed in membrane-bound PhoQD179L(A). These results suggest that PhoQD179L(A) is a locked-on mutant that is defective in extracellular Mg(2+) sensing and that the D179 amino acid residue of PhoQ plays an essential role in signal transfer between the Mg(2+)-sensory and histidine kinase domain of PhoQ. PMID- 16041132 TI - Dose-dependent suppression of tea catechins with a galloyl moiety on postprandial hypertriglyceridemia in rats. AB - Tea has long been believed to be a healthy beverage, and its beneficial effects are almost all attributed to catechins. The effect of catechins on postprandial hypertriglyceridemia in rats was investigated in this study. A lipid emulsion administered orally to rats with (-)-epigallocatechin gallate at a dose of 100 mg/kg resulted in the increase in plasma triacylglycerol being significantly inhibited after 1 and 2 h compared to the case without (-)-epigallocatechin gallate. The effect of (-)-epigallocatechin was weaker than that of (-) epigallocatechin gallate. A tea extract (THEA-FLAN 90S), mainly composed of catechins with a galloyl moiety, dose-dependently suppressed postprandial triacylglycerol after the administration of a lipid emulsion at doses of 50-200 mg/kg. The administration of the tea extract alone at a dose of 200 mg/kg had no effect on the plasma triacylglycerol level. These results strongly suggest that catechins with a galloyl moiety would be promising agents for suppressing dietary fat absorption through the small intestine. PMID- 16041133 TI - Mode of action on deacetylation of acetylated methyl glycoside by cellulose acetate esterase from Neisseria sicca SB. AB - The regioselective deacetylation of purified cellulose acetate esterase from Neisseria sicca SB was investigated on methyl 2,3,4,6-tetra-O-acetyl-beta-D glucopyranoside and 2,3,4,6-tetra-O-acetyl-beta-D-galactopyranoside. The substrates were used as model compounds of cellulose acetate in order to estimate the mechanism for deacetylation of cellulose acetate by the enzyme. The enzyme rapidly deacetylated at position C-3 of methyl 2,3,4,6-tetra-O-acetyl-beta-D glucopyranoside to accumulate 2,4,6-triacetate as the main initial reaction product in about 70% yield. Deacetylation was followed at position C-2, and generated 4,6-diacetate in 50% yield. The enzyme deacetylated the product at positions C-4 and C-6 at slower rates, and generated 4- and 6-monoacetates at a later reaction stage. Finally, it gave a completely deacetylated product. For 2,3,4,6-tetra-O-acetyl-beta-D-galactopyranoside, CA esterase deacetylated at positions C-3 and C-6 to give 2,4,6- and 2,3,4-triacetate. Deacetylation proceeded sequentially at positions C-3 and C-6 to accumulate 2,4-diacetate in 55% yield. The enzyme exhibited regioselectivity for the deacetylation of the acetylglycoside. PMID- 16041134 TI - Suppressive effect of astaxanthin isolated from the Xanthophyllomyces dendrorhous mutant on ethanol-induced gastric mucosal injury in rats. AB - Ethanol has been found to induce ulcerative gastric lesion in humans. The present study investigated the in vivo protective effect of astaxanthin isolated from the Xanthophyllomyces dendrorhous mutant against ethanol-induced gastric mucosal injury in rats. The rats were treated with 80% ethanol for 3 d after pretreatment with two doses of astaxanthin (5 and 25 mg/kg of body weight respectively) for 3 d, while the control rats received only 80% ethanol for 3 d. The oral administration of astaxanthin (5 and 25 mg/kg of body weight) showed significant protection against ethanol-induced gastric lesion and inhibited elevation of the lipid peroxide level in gastric mucosa. In addition, pretreatment with astaxanthin resulted in a significant increase in the activities of radical scavenging enzymes such as superoxide dismutase, catalase, and glutathione peroxidase. A histologic examination clearly indicated that the acute gastric mucosal lesion induced by ethanol nearly disappeared after pretreatment with astaxanthin. PMID- 16041135 TI - Reduction of catechin astringency by the complexation of gallate-type catechins with pectin. AB - The reductive effect of pectin on tea catechin astringency was investigated by using a taste sensor system and (1)H-NMR spectroscopy. The sensor analysis revealed that the astringency of gallate-type catechins (EGCg and ECg) was reduced by the addition of pectin, whereas that of non-gallate-type catechins (EGC and EC) hardly changed. Changes in the (1)H-NMR chemical shifts of the catechins and pectin in mixed solutions showed that the gallate-type catechins formed complexes with pectin more closely than the non-gallate-type catechins. These results demonstrate that complexation between the gallate-type catechins and pectin is a factor for reducing catechin astringency. PMID- 16041136 TI - Antioxidative and antihypertensive effects of Welsh onion on rats fed with a high fat high-sucrose diet. AB - The effects of Welsh onion on the development of hypertension and autoxidation were studied in 6-week-old male Sprague-Dawley rats. The rats were fed with a control diet or a high-fat high-sucrose (HFS) diet with or without 5% Welsh onion (green-leafy type or white-sheath type) for 4 weeks. The systolic blood pressure was elevated and the thiobarbituric acid reactive substances (TBARS) in plasma were increased in the rats fed with the HFS diet without Welsh onion. The rats fed with the HFS diet containing Welsh onion, especially the green-leafy type, had lower blood pressure. They also had a higher level of nitric oxide (NO) metabolites in both the urine and plasma, lower activity of NADH/NADPH oxidase in the aorta, and suppressed angiotensin II production. The effect of white Welsh onion on decreasing the blood pressure was not significant, although the effects on increasing NO metabolites in the urine and decreasing NADH oxidase activity in the aorta were significant. The TBARS value in the plasma was lowered in the rats fed with either green or white Welsh onion, but the in vitro radical scavenging and ferric reducing antioxidative activities were much higher with green Welsh onion than with the white type. These results suggest that the green-leafy Welsh onion, but not the white type, reduced superoxide generation by suppressing the angiotensine II production and then the NADH/NADPH oxidase activity, increasing the NO availability in the aorta, and consequently lowering the blood pressure in the rats fed with the HFS diet. The radical scavenging and reducing antioxidative activities of green Welsh onion may also be effective in decreasing superoxide. PMID- 16041137 TI - Koshikamide A2, a cytotoxic linear undecapeptide isolated from a marine sponge of Theonella sp. AB - Koshikamide A2 (2) was isolated as a cytotoxic metabolite from a marine sponge of Theonella sp. Its structure was elucidated to be a linear undecapeptide by spectroscopic and chemical methods, together with enzymatic conversion to known koshikamide A1 (1). The new peptide moderately inhibited the growth of P388 murine leukemia cells. PMID- 16041138 TI - Odor-active constituents in fresh pineapple (Ananas comosus [L.] Merr.) by quantitative and sensory evaluation. AB - By application of aroma extract dilution analysis (AEDA) to an aroma distillate prepared from fresh pineapple using solvent-assisted flavor evaporation (SAFE), 29 odor-active compounds were detected in the flavor dilution (FD) factor range of 2 to 4,096. Quantitative measurements performed by stable isotope dilution assays (SIDA) and a calculation of odor activity values (OAVs) of 12 selected odorants revealed the following compounds as key odorants in fresh pineapple flavor: 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDF; sweet, pineapple-like, caramel-like), ethyl 2-methylpropanoate (fruity), ethyl 2-methylbutanoate (fruity) followed by methyl 2-methylbutanoate (fruity, apple-like) and 1-(E,Z) 3,5-undecatriene (fresh, pineapple-like). A mixture of these 12 odorants in concentrations equal to those in the fresh pineapple resulted in an odor profile similar to that of the fresh juice. Furthermore, the results of omission tests using the model mixture showed that HDF and ethyl 2-methylbutanoate are character impact odorants in fresh pineapple. PMID- 16041139 TI - Stable isotope dilution-based accurate comparative quantification of nitrogen containing metabolites in Arabidopsis thaliana T87 cells using in vivo (15)N isotope enrichment. AB - Stable isotope dilution-based comparative quantification of nitrogen-containing metabolites for highly sensitive and selective metabolomics was developed using liquid chromatography/mass spectrometry (LC/MS) and (15)N-isotope enrichment. We produced metabolically stable isotope-labeled Arabidopsis T87 cells by culturing with (15)N-labeled medium. We found that the growth of cells maintained in (15)N labeled medium is very similar to the growth in normal medium, as evidenced by cell morphology, doubling time, and measurement of chlorophyll and carotenoid contents. Complete incorporation of (15)N in folate, S-adenosylmethionine (SAM), and S-adenosylhomocysteine (SAH) in T87 cells was accomplished after culturing for 21 d. Accurate comparative quantification of folate, SAM, and SAH was established by means of LC/MS using the isotopomers of the target metabolites as internal standards. The within- and between-run assay coefficients of variation for the folate, SAM, and SAH levels were all less than 8.5%. Stable isotope labeling by nitrogen source in Arabidopsis T87 cell culture provided simple, inexpensive, and accurate amino acid profiling. This interesting new protocol is valuable for the study of dynamic changes in N-compound pools in cultured cells. PMID- 16041140 TI - Localization and interaction of the biosynthetic proteins for the lantibiotic, Nukacin ISK-1. AB - Nukacin ISK-1 is a type-A(II) lantibiotic produced by Staphylococcus warneri ISK 1. In this study, we characterized NukM and NukT, which are predicted to be involved in modification of prepeptide (NukA) and cleavage of leader peptide and subsequent secretion respectively. Localization analysis of NukM and NukT in the wild-type strain indicated that both proteins were located at the cytoplasm membrane. Interestingly, NukM expressed heterologously in St. carnosus TM300 was also located at the cytoplasm membrane even in the absence of NukT. Yeast two hybrid assay showed that a complex of at least two each of NukM and NukT was associated with NukA. In vitro interaction analysis by surface plasmon resonance biosensor further suggested that membrane-located NukM interacted with NukA. These results indicate that NukM and NukT form a membrane-located multimeric protein complex and that post-translational modification of nukacin ISK-1 would occur at the cytoplasm membrane. PMID- 16041141 TI - Concise synthesis of (8Z,11Z,14Z)-8,11,14-heptadecatrienal, (7Z,10Z,13Z)-7,10,13 hexadecatrienal, and (8Z,11Z)-8,11-heptadecadienal, components of the essential oil of marine green alga Ulva pertusa. AB - The long-chain aldehydes, (8Z,11Z,14Z)-8,11,14-heptadecatrienal, (7Z,10Z,13Z) 7,10,13-hexadecatrienal, and (8Z,11Z)-8,11-heptadecadienal, were concisely synthesized by using Grignard coupling, catalytic hydrogenation with the Lindlar catalyst, and oxidation with Dess-Martin periodinane as the key steps. Particularly, (8Z,11Z,14Z)-8,11,14-heptadecatrienal and (7Z,10Z,13Z)-7,10,13 hexadecatrienal both possessed a seaweed-like odor. PMID- 16041142 TI - Glucose trimming of N-glycan in endoplasmic reticulum is indispensable for the growth of Raphanus sativus seedling (kaiware radish). AB - Recently I found that glycosidase inhibitors such as castanospermine, deoxynojirimycin, swainsonine, 2-acetamindo 2,3-dideoxynojirimycin, and deoxymannojirimycin change the N-glycan structure of root glycoproteins, and that the glucosidase inhibitors castanospermine and deoxynojirimycin suppress the growth of Raphanus sativus seedlings (Mega, T., J. Biochem., 2004). The present study undertook to see whether the growth suppression is due to the inhibition of glucose trimming in endoplasmic reticulum (ER). The study, using three glucosidase inhibitors, castanospermine, N-methyl deoxynojirimycin, and deoxynojirimycin, upon the growth of R. sativus foliage leaf, made clear that glucose trimming is indispensable for plant growth, because the inhibition of glucose trimming correlated with leaf growth. On the other hand, processing inhibition in the Golgi apparatus by other glycosidase inhibitors had little effect on plant growth, although N-glycan processing was disrupted depending on inhibitor specificity. These results suggest that N-glycan processing after glucosidase processing is dispensable for plant growth and cell differentiation. PMID- 16041143 TI - Effect of high-pressure gas on yeast growth. AB - Microcalorimetry is a useful tool for monitoring the growth behavior of microorganisms. In this study, microcalorimetry was used to investigate the effects of nitrogen, air, oxygen, nitrous oxide, argon, and krypton at high pressure on the growth of the yeast Saccharomyces cerevisiae. Growth thermograms (metabolic heat vs. incubation time) were generated to estimate metabolic activity under compressed gases and to determine the 50% inhibitory pressure (IP(50)) and minimum inhibitory pressure (MIP), which are regarded as indices of the toxicity of compressed gases. Based on MIP values, the most toxic to the least toxic gases were found to be: O(2) > N(2)O > air > Kr > N(2) > Ar. PMID- 16041144 TI - Construction of a bacterial artificial chromosome library for a myxobacterium of the genus Cystobacter and characterization of an antibiotic biosynthetic gene cluster. AB - A bacterial artificial chromosome (BAC) library was constructed to isolate the biosynthetic gene cluster for the polyketide/peptide hybrid-type antibiotic cystothiazole A from the myxobacterium Cystobacter fuscus strain AJ-13278. Sequence analysis of a 63.9 kb contiguous region that encompasses the biosynthetic gene cluster (cta) led to the identification of a polyketide synthase (PKS)/nonribosomal peptide synthetase (NRPS) hybrid gene cluster 32.1 kb in size, which consists of six open reading frames (ORFs), ctaB to ctaG, as well as downstream genes ctaJ and ctaK (1.0 and 0.9 kb, respectively) responsible for the final biosynthetic steps. The genes ctaB, ctaE, and ctaF encode PKSs, the genes ctaC and ctaG encode NRPSs, and ctaD encodes an NRPS-PKS hybrid enzyme. Disruption of ctaD impaired cystothiazole A production. Additionally, two downstream genes, ctaJ and ctaK, which encode a nitrilase and an O methyltransferase, respectively, must be responsible for the final methyl ester formation in the cystothiazole A biosynthesis. PMID- 16041145 TI - Reverse reaction of Aspergillus niger APC-9319 alpha-galactosidase in a supersaturated substrate solution: production of alpha-linked galactooligosaccharide (alpha-GOS). AB - The alpha-galactosidase that effectively catalyzes a reverse reaction of galactose, Aspergillus niger APC-9319 alpha-galactosidase, was screened from industrial enzyme preparations for food processing containing alpha-galactosidase activity. Reverse reaction of A. niger APC-9319 alpha-galactosidase was performed using a supersaturated solution (90% galactose [w/v]). A. niger APC-9319 alpha galactosidase was not inhibited even in high substrate concentration, and effectively catalyzed the reverse reaction. The yield of the reaction product, alpha-linked galactooligosaccharide (alpha-GOS), increased greatly as the initial concentration of galactose increased to 90% (w/v), and was more than 50%. Furthermore, the half life of enzyme activity was about three times as long as that using 60% galactose (w/v). alpha-GOS (1.4 g) was prepared from galactose (3.0 g) by reverse reaction of A. niger APC-9319 alpha-galactosidase. The alpha GOS contained 58% alpha-galactobiose (alpha-Gal2), 28% alpha-galactotriose, and 14% oligosaccharides larger than alpha-galactotriose. The main component of positional isomers in alpha-Gal2 was alpha-1,6Gal2. PMID- 16041146 TI - Identification of aminotransferase genes for biosynthesis of aminoglycoside antibiotics from soil DNA. AB - Aminoglycoside has been known as a clinically important antibiotic for a long time, but genetic information for the biosynthesis of aminoglycoside is still insufficient. In this study, we tried to clone aminoglycoside-biosynthetic genes from soil DNA for accumulation of genetic information. We chose the genes encoding L-glutamine:(2-deoxy-)scyllo-inosose aminotransferase as the target, because it is specific for all types of aminoglycoside biosynthesis. By degenerate PCR, we obtained 33 individual clones that were homologous with aminotransferase genes in aminoglycoside biosynthesis. Phylogenetic analysis and alignment of these genes showed that horizontal gene transfer has occurred in the soil. Among these, several quite interesting genes were obtained. Some genes probably originated from non-actinomycetes, and some were far from the known homologs. These genes can be useful markers for the isolation of entire gene clusters and originating organisms. PMID- 16041147 TI - Functional dissection of a mercuric ion transporter, MerC, from Acidithiobacillus ferrooxidans. AB - Topological analysis with a phoA gene fusion suggested that Acidithiobacillus ferrooxidans MerC, a mercury transporter, has two periplasmic loops and four transmembrane domains. Cys-23 and Cys-26 of the protein were involved in Hg(2+) recognition/uptake, but Cys-132 and Cys-137 were not. Escherichia coli cells producing the MerC were hypersensitive to CdCl(2). In this case, mutation of His72 rendered the host cells less CdCl(2) sensitive, whereas none of the Cys residues affected it. E. coli cells expressing the gene encoding a mercuric ion transporter (merC)-deletion mutant, in which the coding-sequence of the carboxy terminal cytoplasmic region was removed, retained Hg(2+) hypersensitivity and showed about 55% HgCl(2) uptake ability compared to that of the one expressing the intact merC, indicating that the region is not essential for Hg(2+) uptake. Coexpression of A. ferrooxidans the gene encoding mercuric reductase (merA) and the merC deletion mutation conferred HgCl(2) tolerance to E. coli host cells. Under this condition, the merC deletion gene product was exclusively present as a monomer. PMID- 16041148 TI - A novel type of immunity protein, NukH, for the lantibiotic nukacin ISK-1 produced by Staphylococcus warneri ISK-1. AB - Staphylococcus warneri ISK-1 produces a lantibiotic, nukacin ISK-1. The nukacin ISK-1 gene cluster consists of at least six genes, nukA, -M, -T, -F, -E, and -G, and two open reading frames, ORF1 and ORF7 (designated nukH). Sequence comparisons suggested that NukF, -E, -G, and -H contribute to immunity to nukacin ISK-1. We investigated the immunity levels of recombinant Lactococcus lactis expressing nukFEG and nukH against nukacin ISK-1. The co-expression of nukFEG and nukH resulted in a high degree of immunity. The expression of either nukFEG or nukH conferred partial immunity against nukacin ISK-1. These results suggest that NukH contributes cooperatively to self-protection with NukFEG. The nukacin ISK-1 immunity system might function against another lantibiotic, lacticin 481. Western blot analysis showed that NukH expressed in Staphylococcus carnosus was localized in the membrane. Peptide release/bind assays indicated that the recombinant L. lactis expressing nukH interacted with nukacin ISK-1 and lacticin 481 but not with nisin A. These findings suggest that NukH contributes cooperatively to host immunity as a novel type of lantibiotic-binding immunity protein with NukFEG. PMID- 16041149 TI - Molecular cloning and expression analysis of two hepcidin genes from olive flounder Paralichthys olivaceus. AB - Hepcidin is a cysteine-rich cationic antimicrobial peptide central to iron metabolism. We report a comparative analysis of the sequences, gene organization and expression of two hepcidin genes from olive flounder Paralichthys olivaceus. Both consist of two introns and three exons that encode a prepropeptide (81 amino acids for hepcidin I and 89 amino acids for hepcidin II). A TATA box and several consensus-binding motifs for transcription factors were found upstream of the transcriptional starting site. Hepcidin II was predominantly expressed in the liver and highly inducible under the effect of lipopolysaccharide (LPS), while a large amount of hepcidin I transcripts was detected in various tissues but did not appear to have a significant effect during LPS-stimulation. PMID- 16041150 TI - Protein-DNA interactions under high-pressure conditions, studied by capillary narrow-tube electrophoresis. AB - The method of electrophoretic mobility shift assay under high-pressure conditions was improved using a high-pressure electrophoresis apparatus with capillary narrow-tube gel. It was found that the protein-DNA complex in the gel was stained as a high-resolution spot with ethidium bromide. Using this method, it was found that the behavior under high-pressure conditions of the protein-DNA complex composed of NtrC protein and its target promoter DNA is important for the pressure-regulated transcription process, and it was confirmed that the complex was dissociated above a pressure of 70 MPa. PMID- 16041151 TI - Unexpected elevated production of Aquifex aeolicus cytochrome c555 in Escherichia coli cells lacking disulfide oxidoreductases. AB - Mutant strains of Escherichia coli lacking DsbA, DsbB, or DsbD (proteins required for disulfide bond formation in the periplasm) did not produce mitochondrial or chloroplast cytochromes c, as previously observed for bacterial ones. Unexpectedly, however, cytochrome c(555) (AA c(555)) from a hyperthermophile, Aquifex aeolicus, was produced in the E. coli periplasm without Dsb proteins, three times more than with them. These results indicate that the Dsb proteins are not necessarily required for AA c(555) production in E. coli, possibly because of hyperthermophilic origin compared with the others. PMID- 16041152 TI - Functional conservation between fission yeast moc1/sds23 and its two orthologs, budding yeast SDS23 and SDS24, and phenotypic differences in their disruptants. AB - The moc1/sds23 gene was isolated to induce sexual development of a sterile strain due to overexpression of adenylate cyclase in Schizosaccharomyces pombe. Here, we studied the functional conservation between moc1/sds23 and its two orthologs SDS23 and SDS24 in Saccharomyces cerevisiae. We observed that the temperature sensitivity, salt tolerance, cell morphology, and sterility of the Deltamoc1 mutant in S. pombe were recovered by expressing either S. cerevisiae SDS23 or SDS24. We found that deletion of both SDS23 and SDS24 resulted in the production of a large vacuole that was reversed by the expression of S. pombe moc1/sds23. In these ways we found that S. pombe Moc1/Sds23 and S. cerevisiae SDS23p or SDS24p are functional homologs. In addition we found that the Deltasds23 Deltasds24 diploid strain reduces cell separation in forming pseudohyphal-like growth in S. cerevisiae. Thus S. pombe moc1/sds23 and S. cerevisiae SDS23 or SDS24 are interchangeable with each other, but their disruptants are phenotypically dissimilar. PMID- 16041153 TI - Three-year follow-up results of angiographic intervention trial using an HMG-CoA reductase inhibitor to evaluate retardation of obstructive multiple atheroma (ATHEROMA) study. AB - BACKGROUND: The cardiovascular event rate in Japanese populations is strikingly lower than that in Caucasian populations and it has been postulated that this difference might be because of variations in atherosclerosis progression in patients with coronary artery disease (CAD). However, the rate of angiographically assessed progression and its response to statins has not been well described in Japanese patients. METHODS AND RESULTS: The angiographic intervention trial using an HMG-CoA reductase inhibitor to evaluate the retardation of obstructive multiple atheroma (ATHEROMA) study was a multicenter, randomized, controlled clinical trial investigating the effects of pravastatin on coronary atherosclerosis in Japanese patients with CAD using quantitative coronary angiography. In total, 361 patients with mild to moderate elevated serum total cholesterol concentrations (195-265 mg/dl) received diet only (n = 179) or diet plus pravastatin 10-20 mg/dl (n = 182). Over 3 years, low-density lipoprotein-cholesterol in the pravastatin group decreased by 19.5% (p < 0.0001). A per-patient analysis showed that minimum lumen diameter increased by 0.034+/ 0.17 mm in the pravastatin group, but decreased by 0.006+/-0.16 mm in the diet only group (p = 0.04). The mean difference between the treatment groups was 0.040 mm (95% confidence interval 0.020, 0.070 mm). CONCLUSION: The ATHEROMA study indicates that pravastatin 10-20 mg/day for 3 years improves hyperlipidemia, then suppresses progression and induces regression of focal coronary atherosclerosis in Japanese CAD patients with elevated serum cholesterol. PMID- 16041154 TI - Reperfusion phenomenon is a strong predictor of left ventricular remodeling after acute myocardial infarction. AB - BACKGROUND: Many clinicians have seen the reperfusion phenomenon, a paradoxical response that includes a transient increase of chest pain, additional ST-segment elevation or ventricular arrhythmias immediately after coronary reperfusion, in patients with acute myocardial infarction (AMI). The aim of the present study was to investigate the impact of this phenomenon during coronary reperfusion on left ventricular (LV) remodeling in patients with AMI. METHODS AND RESULTS: One hundred and thirty-eight consecutive patients with a first anterior-wall AMI, undergoing coronary reperfusion treatment within 24 h of onset were prospectively evaluated for reperfusion phenomenon and followed up with scheduled evaluations of LV function and morphology with left ventriculography for 1 year. Of the 138 enrolled patients, 77 underwent serial left ventriculography at the acute, subacute and 1-year phases. Of these 77 patients, 39 demonstrated the reperfusion phenomenon. The LV end-diastolic volume index significantly increased from the acute to subacute phase and to the 1-year phase, but was unchanged in the 38 patients without reperfusion phenomenon. In multivariate analysis, reperfusion phenomenon was the only determinant of LV dilatation after AMI. CONCLUSIONS: Reperfusion phenomenon was a strong predictor of LV remodeling after reperfusion therapy for AMI. PMID- 16041155 TI - Serial changes in circulating concentrations of soluble CD40 ligand and C reactive protein in patients with unstable angina undergoing coronary stenting. AB - BACKGROUND: This study tested the hypothesis that serum concentrations of high sensitivity C-reactive protein (hs-CRP) and soluble CD40 ligand (sCD40L) significantly reflect serial changes in patients with unstable angina, and thus the serum concentrations of these inflammatory biomarkers may be good candidates for predicting late restenosis after coronary stenting. METHODS AND RESULTS: The circulating concentrations of sCD40L and hs-CRP were prospectively measured (both pre-procedure, and on days 21, 90, and 180 after the procedure) in 77 consecutive patients with unstable angina undergoing coronary stenting. These inflammatory mediators were also evaluated in 30 healthy volunteers. The serum concentrations of sCD40L and hs-CRP were significantly higher pre-procedure in study patients than in normal control subjects (all p values < 0.0001). These inflammatory markers then declined to a substantially lower concentration by day 21 (all p values < 0.05). Circulating concentrations of hs-CRP in each patient then differed little from each other afterwards. However, the sCD40L concentration was once again raised significantly on days 90 and 180 as compared to day 21 (both p values < 0.05). This study found no significant link between raised circulating concentrations of sCD40L and hs-CRP and late restenosis. CONCLUSIONS: Circulating concentrations of sCD40L and hs-CRP were significantly increased in unstable angina patients pre-procedure and declined substantially thereafter. However, the circulating concentrations of these 2 inflammatory mediators were not useful in predicting late restenosis following coronary stenting. PMID- 16041156 TI - Serial angiographic follow-up beyond 10 years after coronary artery bypass grafting. AB - BACKGROUND: The long-term prognosis and serial angiographic follow-up beyond 10 years in patients who underwent coronary artery bypass grafting (CABG) have not been fully studied in Japan. METHODS AND RESULTS: In the present study data from 71 patients who underwent CABG before 1992 were analyzed. Thirty patients had a saphenous vein graft (SVG) only group, and the remaining 41 had a left internal thoracic artery graft to the left anterior descending coronary artery (LITA) group; 6 patients died from malignancy, which was the most common cause of death after CABG. The major adverse cardiac events (MACE) were defined as cardiac death, Q-wave or nonQ-wave myocardial infarction, and congestive heart failure. The MACE-free rate was significantly higher in the LITA group than in the SVG group (p < 0.05). However, among the patients with an ejection fraction < 0.40, there was no significant difference in MACE-free rate between the 2 groups. The LITA patency rate was significantly higher than that for SVG (p < 0.05) and the SVG patency rate was lower in the patients with hyperlipidemia (p < 0.05); cholesterol-lowering therapy improved the SVG patency rate. CONCLUSION: The long term outcome of CABG was favorable, particularly if using an arterial graft. Although the patency rate was lower for the SVG than LITA, the patient's lipid profile might be an important factor in the SVG patency rate. PMID- 16041157 TI - Assessment of coronary intima--media thickness by optical coherence tomography: comparison with intravascular ultrasound. AB - BACKGROUND: Optical coherence tomography (OCT) is a new imaging method. With a resolution of approximately 10-20 Em, which is approximately 10-fold higher than that of intravascular ultrasound (IVUS). METHODS AND RESULTS: This study compared the coronary intima - media thickness (IMT) and the intimal thickness of 54 coronary arterial segments evaluated by histological examination with the results of OCT and IVUS. There was better agreement in IMT between OCT and histological examination than between IVUS and histological examination (r = 0.95, p < 0.001, mean difference = -0.01+/-0.07 mm for OCT; r = 0.88, p < 0.001, mean difference = -0.03+/-0.10 mm for IVUS). There was good agreement in the intimal thickness between OCT and histological examination (r = 0.98, p < 0.001, mean difference = 0.01+/-0.04 mm). CONCLUSIONS: IMT could be measured more accurately by OCT than IVUS. In addition, the intimal thickness could be evaluated by OCT and correlated well with the histological examination. PMID- 16041158 TI - Intracoronary continuous adenosine infusion. AB - BACKGROUND: Various methods are used to induce maximal hyperemia for physiologic studies, but the feasibility and efficacy of continuous intracoronary (IC) infusion of adenosine for measurement of fractional flow reserve (FFR) has not been well-defined. METHODS AND RESULTS: Patients with intermediate coronary artery stenosis were consecutively enrolled. In the phase I study, FFR was measured after 3 dosages of IC adenosine infusion (180, 240 and 300 microg/min) in 30 patients. The phase II study was performed to compare the hyperemic efficacy of IC infusion (240 microg/min) with IC bolus injection (40, 80 microg) and intravenous (IV) infusion (140 microg x kg (-1) x min(-1)) of adenosine in 20 patients. In the phase I study, no significant differences in FFR were observed with the 3 different doses of IC infusion (p = 0.06). In the phase II study, FFR after an IC bolus injection (0.83+/-0.06) was significantly higher than with IV (0.79+/-0.07) or IC (0.78+/-0.09) infusion (p < 0.01). However, no difference in FFR was observed for IC and IV infusions. CONCLUSION: IC infusion of adenosine seems to be a safe and effective method of inducing maximal hyperemia for FFR measurement. PMID- 16041159 TI - Efficacy of nasal bi-level positive airway pressure in congestive heart failure patients with cheyne-stokes respiration and central sleep apnea. AB - BACKGROUND: Cheyne - Stokes respiration with central sleep apnea (CSR-CSA) contributes to the poor prognosis in patients with congestive heart failure (CHF). Bi-level positive airway pressure (bi-level PAP) may be an effective alternative for treating CSR-CSA and CHF. METHODS AND RESULTS: Fourteen patients with CSR-CSA were divided into 2 groups, a control group that included 7 patients who decided to receive only conventional medications and a group of 7 patients that received bi-level PAP. Left ventricular ejection fraction (LVEF), mitral regurgitation (MR) area, plasma brain natriuretic peptide (BNP) concentration and the New York Heart Association (NYHA) functional class were evaluated initially (baseline) and 3 months later. In the control group, there were no significant changes in cardiac function during the study period. In contrast, in the group that received bi-level PAP, there were significant improvements in LVEF (from 36.3+/-2.9% to 46.0+/-4.0%, p = 0.02), MR area (from 30.4+/-7.6% to 20.0+/-5.1%, p = 0.02), BNP (from 993.6+/-332.0 pg/ml to 474.0+/-257.6 pg/ml, p = 0.02) and NYHA functional class (from 3.1+/-0.1 to 2.1+/-0.1, p = 0.03). CONCLUSION: Treatment with bi-level PAP improved cardiac functions in CHF patients with CSR CSA. PMID- 16041160 TI - Combination of heart-type fatty acid binding protein and brain natriuretic peptide can reliably risk stratify patients hospitalized for chronic heart failure. AB - BACKGROUND: The aim of the present study was to prospectively study whether a combination of markers for myocardial cell injury and left ventricular overload at admission can reliably risk stratify patients hospitalized for chronic heart failure (CHF). METHODS AND RESULTS: Serum concentrations of heart-type fatty acid binding protein (H-FABP) and plasma concentrations of brain natriuretic peptide (BNP) were measured at admission in 186 consecutive patients hospitalized for CHF. During a mean follow-up period of 534+/-350 days, there were 44 cardiac events, including 16 cardiac deaths and 28 readmissions for worsening heart failure. Normal upper limits for H-FABP and BNP values were determined from the receiver operating characteristic curves (4.3 ng/ml for H-FABP and 200 pg/ml for BNP). A stepwise Cox regression analysis demonstrated that high H-FABP (hazard ratio 5.416, p = 0.0002) and high BNP (hazard ratio 2.411, p = 0.0463) were independent predictors of cardiac events. High concentrations of both H-FABP and BNP at admission were associated with the highest incidence of cardiac mortality and cardiac events. Kaplan-Meier analysis also showed that the combination of H FABP and BNP concentrations could reliably stratify patients for cardiac events. CONCLUSION: Combined measurement of H-FABP and BNP concentrations at admission may be a highly reliable evaluation for risk stratifying patients hospitalized for CHF. PMID- 16041161 TI - Relationship between serum uric acid concentration and insulin resistance and metabolic syndrome. AB - BACKGROUND: Associations between hyperuricemia, cardiovascular diseases and diabetes have been reported, but few of the studies have been conducted in the Korean population. The present study examined a Korean adult population with respect to the relationships between serum uric acid concentrations and hypertension, insulin resistance, and the risk factors of metabolic syndrome. METHODS AND RESULTS: A total of 53,477 subjects were divided into 4 groups according to serum uric acid quartiles. The incidence of hypertension in all subjects was higher in the first quartile than in the third plus fourth quartile (odds ratio (OR) 1.192, p < 0.001). Homeostasis model assessment index was found to be associated with serum uric acid concentration in all subjects (OR 1.193, p < 0.001), and the serum uric acid concentration was positively correlated with the risk factors of metabolic syndrome. In addition, the number of metabolic syndrome variables increased as serum uric acid concentration increased. CONCLUSIONS: Serum uric acid concentration was found to be independently correlated with hypertension, insulin resistance and the risk factors of metabolic syndrome. In addition, even those with a serum uric acid concentration in the normal range showed an increased risk of metabolic syndrome as serum uric acid concentration increased. PMID- 16041162 TI - Assessment of coronary microcirculation in patients with takotsubo-like left ventricular dysfunction. AB - BACKGROUND: The precise mechanism of takotsubo-like left ventricular (LV) dysfunction remains unclear. METHODS AND RESULTS: Eight consecutive patients with takotsubo-like LV dysfunction were studied. In the acute phase and 3 weeks later, the coronary flow velocity spectrum and coronary flow velocity reserve (CFVR) were recorded by Doppler guidewire. The deceleration time of diastolic velocity (DDT; ms) was measured from the peak diastolic velocity to the point where the extrapolated line intersected the baseline. In all cases, CFVR of the 3 coronary arteries increased during follow-up (1.7+/-0.5 to 2.4+/-0.5 for the left anterior descending artery (p < 0.05), 1.7+/-0.5 to 2.7+/-0.8 for the left circumflex artery (p < 0.05) and 1.7+/-0.3 to 2.5+/-0.4 for the right coronary artery (p < 0.05)). In addition, the DDT of all coronary arteries increased during follow-up. CONCLUSIONS: Decreased CFVR and short DDT were demonstrated in patients with takotsubo-like transient LV dysfunction in the acute phase, and these findings improved 3 weeks later, which suggests that coronary microvascular dysfunction may be a causative mechanism. PMID- 16041163 TI - Antiarrhythmic drug cibenzoline attenuates left ventricular pressure gradient and improves transmitral Doppler flow pattern in patients with hypertrophic obstructive cardiomyopathy caused by midventricular obstruction. AB - BACKGROUND: Recent interventional and surgical therapies to attenuate left ventricular pressure gradient (LVPG) can be difficult to perform in patients with hypertrophic obstructive cardiomyopathy (HOCM) caused by midventricular obstruction (MVO), owing to the risk of inducing or deteriorating mitral regurgitation. METHODS AND RESULTS: The effects of the antiarrhythmic drug, cibenzoline, on LVPG and left ventricular (LV) diastolic function estimated by the change in the transmitral Doppler flow pattern were examined in 23 patients with HOCM and MVO. Hemodynamic changes 2 h after a single dose of 200 mg of cibenzoline and 3 months after oral administration of 300-450 mg of cibenzoline per day were examined. At 2 h after the treatment, LVPG decreased from 79+/-37 mmHg to 24+/-21 mmHg (p < 0.0001). E-wave velocity significantly increased and A wave velocity significantly decreased, and thus the E/A ratio increased from 0.83+/-0.39 to 1.36+/-0.50 (p < 0.0001). After 3 months of treatment, LVPG remained decreased, and the E-wave and A-wave velocities and the E/A ratio remained improved. CONCLUSIONS: Cibenzoline can attenuate LVPG and ameliorate LV diastolic dysfunction in patients with HOCM caused by MVO, which suggests a new strategy for the management of this condition. PMID- 16041164 TI - Sinus node dysfunction concomitant with Brugada syndrome. AB - BACKGROUND: A genetic correlation between Brugada syndrome (BS) and sinus node dysfunction (SND) has been proposed, although the clinical and electrophysiologic characteristics of this concomitant condition are unknown. METHODS AND RESULTS: The study comprised 5 patients with symptomatic BS (4 with spontaneous episodes of ventricular fibrillation (VF) and 1 with syncope) of whom 3 had a documented sinus pause > 3 s (a 42- and 62-year-old man, and a 49-year-old woman). Only 1 of them had a family history of sudden death; 2 of them had also had an episode of atrial fibrillation or flutter. Electrophysiologic study demonstrated prolonged sinus node recovery time in 2 patients (2.6 s and > 5 s), in whom a cardiac pacemaker had been implanted before the diagnosis of BS was made after episodes of VF. Finally, all 3 patients received an implantable cardioverter defibrillator, including 2 upgrades from pacemaker. CONCLUSIONS: SND is not a rare concomitant disorder in BS and there is a possible genetic connection. PMID- 16041165 TI - Functional assessment by myocardial performance index (Tei index) correlates with plasma brain natriuretic peptide concentration in patients with hypertrophic cardiomyopathy. AB - BACKGROUND: Hypertrophic cardiomyopathy (HCM) might exhibit not only diastolic, but also latent systolic dysfunction. Therefore combined assessment of both systolic and diastolic function using myocardial performance index (Tei index) can be useful in HCM. Plasma brain natriuretic peptide (BNP) level is reported to be elevated in HCM, but the mechanism of BNP elevation in HCM remains to be established. METHODS AND RESULTS: The value of Tei index in 45 HCM patients was compared with that of 20 normal control subjects. The HCM patients showed a higher value of Tei index (0.55+/-0.12 vs 0.36+/-0.08, p < 0.0001) and longer isovolumic relaxation and contraction times than control subjects. The plasma BNP level correlated with Tei index in non-obstructive HCM (n = 35, r = 0.61, p < 0.0001), although the correlation was mild when overall HCM patients were included (r = 0.34, p = 0.02). The correlation was still significant after adjusting for age, or the extent and severity of left ventricular hypertrophy. Multiple stepwise regression analysis identified mitral E/A ratio (r = 0.49, F = 13.1) and Tei index (r = 0.37, F = 7.6) as independent predictors of higher plasma BNP level in non-obstructive HCM. CONCLUSIONS: Myocardial performance index was abnormal in HCM, reflecting both systolic and diastolic dysfunction in this disorder. Plasma BNP level correlated with functional assessment by Tei index in non-obstructive HCM. PMID- 16041166 TI - Serum heart-type fatty acid-binding protein concentration positively correlates with the length of aortic dissection. AB - BACKGROUND: Patients with aortic dissection (AD) often demonstrate positive heart type fatty acid-binding protein (H-FABP), but its significance is unclear. METHODS AND RESULTS: In 63 of 64 consecutive AD patients, the serum H-FABP concentration was measured and the H-FABP positive rate calculated (cutoff value: 6.2 ng/ml) for each of following factors: (1) with or without dissection of the ascending aorta; and (2) a thrombosed false lumen; (3) length score; (4) presence of shock; (5) malperfusion of limbs; (6) ST elevation and/or depression on electrocardiogram; and (7) renal dysfunction. In total 36 AD patients had a positive H-FABP test. Statistically significant differences in the H-FABP positive rate were observed between those with and without ascending AD (76.7% vs 39.4%, p = 0.003), and in the length score (p < 0.001). Multivariate logistic regression test demonstrated that the most significant factor was the length score (odds ratio: 2.239 (95% confidence interval: 1.119-4.481), p = 0.023). Moreover, a significant correlation was observed between length score and absolute H-FABP value (r = 0.420, p = 0.001). CONCLUSIONS: In patients with AD, an increased serum H-FABP concentration is caused by the protein being released not only from the cardiac muscle but also from skeletal muscle, or possibly the aortic wall. Physicians using H-FABP for detection of myocardial injury need to be aware that patients with a long or ascending AD will show an elevation of H FABP. PMID- 16041167 TI - Single high-dose intravenous immunoglobulin therapy for kawasaki disease increases plasma viscosity. AB - BACKGROUND: Intravenous immunoglobulin therapy, widely used for various autoimmune and systemic inflammatory diseases including Kawasaki disease (KD), is occasionally associated with thromboembolic adverse effects caused by an abrupt increase in blood viscosity. Scarce information is available, however, regarding the effect of single high-dose immunoglobulin therapy for KD on blood viscosity. METHODS AND RESULTS: Eleven boys and 5 girls (mean age: 2.1 years) with acute phase KD underwent single high-dose immunoglobulin therapy. Plasma viscosity before the treatment was 1.18 centipoises (SD = 0.06), but it significantly rose to 1.34 centipoises (SD = 0.06) (p < 0.001). Multiple regression analysis revealed that, among various factors including hematocrit, plasma concentrations of total protein, immunoglobulin G (IgG), immunoglobulin A (IgA), and immunoglobulin M (IgM), only plasma IgG concentration was included in the model to explain plasma viscosity (R2 = 0.59, p < 0.001). CONCLUSIONS: Single high-dose regimen for acute-phase KD increases blood viscosity and therefore might increase the risk of thromboembolism. PMID- 16041168 TI - Effect of angiotensin blockade on the association between albuminuria and peripheral arterial disease in elderly Taiwanese patients with type 2 diabetes mellitus. AB - BACKGROUND: The aim of the present study was to examine the impact of the clinical use of angiotensin-converting enzyme inhibitors (ACEI) or angiotensin receptor blockers (ARB) on the association between the urinary albumin/creatinine ratio (ACR) and ankle-brachial index (ABI) or peripheral arterial disease (PAD) in elderly Taiwanese patients with type 2 diabetes mellitus. METHODS AND RESULTS: Two-hundred and ninety patients (108 men, 182 women) aged > or = 65 years (mean +/- SD, 71.6 +/-4.9) were cross-sectionally studied. ACR was expressed as the natural logarithm [ln(ACR)] and divided into normoalbuminuria (< 30.0 microg/mg) and albuminuria (> or = 30.0 microg/mg). ABI was evaluated both continuously and as peripheral arterial disease (PAD) (-) and PAD (+) using the cutoff of 0.9. Statistical analyses were performed with consideration of covariates and the use of ACEI/ARB. Results showed that in patients not using ACEI/ARB, ln (ACR) negatively correlated with ABI (r = -0.261, p < 0.01) and was associated with ABI with adjusted regression coefficient of -0.0213 (p < 0.05). PAD patients had a significantly higher level of ln (ACR) than those without PAD (4.83+/-1.34 vs 3.73+/-1.29, p < 0.001) and PAD prevalence was significantly higher in those with albuminuria than in those with normoalbuminuria (22.6% vs 4.9%, p < 0.001). The multivariate-adjusted odds ratio for PAD for every 1 unit increment of ln (ACR) was 2.10 (1.31-3.38), and for albuminuria vs normoalbuminuria 3.86 (1.04-14.31) in patients not using ACEI/ARB. In patients using ACEI/ARB, none of these analyses was significant. Patients using ACEI/ARB had a significantly lower risk of PAD with a multivariate-adjusted odds ratio of 0.34 (0.12-0.97). CONCLUSIONS: ACR negatively correlates with ABI and is associated with PAD in elderly diabetic patients not treated with ACEI/ARB. The use of ACEI/ARB attenuates this association and may be associated with a lower risk of PAD. PMID- 16041169 TI - Critical analysis of hemodynamic insufficiency by head-up tilt in patients with carotid occlusive disease. AB - BACKGROUND: The objective of this study was to evaluate the diagnostic value of the head-up-tilt (HUT) test for detecting cerebral hemodynamic insufficiency in patients with major cerebral artery occlusion disease because such patients may benefit from extracranial - intracranial bypass surgery. METHODS AND RESULTS: In 13 cases of transient ischemic attacks in patients with carotid or middle cerebral artery occlusive disease, the HUT test was used to determine whether or not the symptoms appear during induced hypotension before investigating cerebral hemodynamics with positron emission tomography. Three of the 13 patients showed focal symptoms such as hemiparesis and limb shaking during the HUT test. In all 3 patients, the oxygen extraction fraction (OEF) increased beyond 53.3% (ie, misery perfusion), whereas only 2 of the other 10 patients without focal symptoms showed an increase in OEF during HUT. CONCLUSIONS: The HUT test was highly useful for screening patients with cerebral hemodynamic insufficiency in carotid occlusive disease. PMID- 16041170 TI - Cardiovagal response to acute mild exercise in young healthy subjects. AB - BACKGROUND: The aim of the present study was to investigate the effect of a single bout of mild exercise on autonomic nervous system activity in healthy subjects. METHODS AND RESULTS: The study group comprised 18 healthy males, aged between 20 and 24 years, who had not been training regularly for the last 3 months. A supine recording of systolic arterial pressure (SAP) and RR interval and the administration of the phenylephrine test were performed at baseline and repeated after a 60-min recovery period following treadmill exercise training for 30 min at 65% of maximal heart rate. Mean SAP and RR interval, heart rate variability (HRV) indices (the standard deviation of normal-to-normal RR intervals (SDNN), the square root of the mean of squared differences between successive intervals and the percentage of adjacent RR intervals differing more than 50 ms), noninvasive spectral baroreflex sensitivity (Spe-BRS) and phenylephrine baroreflex sensitivity (Phe-BRS) were assessed before and after training. Mean SAP measured after exercise was lower than baseline (120+/-12 mmHg vs 128+/-12 mmHg, p = 0.05). Spe-BRS and Phe-BRS increased significantly after exercise, from 11.8+/-6.1 ms/mmHg to 16.0+/-7.8 ms/mmHg (p = 0.034), and from 16.0+/-8.8 ms/mmHg to 21.9+/-9.3 ms/mmHg (p = 0.022), respectively. A parallel increase was also observed in SDNN (from 81+/-44 ms to 96+/-53 ms, p = 0.02), but the other HRV indices showed no significant differences between pre- and post exercise. CONCLUSIONS: A single session of mild exercise performed by sedentary young men leads to significant autonomic nervous system improvement, which suggests that even mild physical activity is beneficial for neural cardiac regulation and should be recommended to sedentary healthy subjects. PMID- 16041171 TI - Differences in clinical presentation of pulmonary embolism in older vs younger patients. AB - BACKGROUND: In order to better define the clinical characterization of pulmonary embolism (PE) in the elderly, the clinical and laboratory findings were compared in older (> or = 65 years old) and younger (< 65 years old) patients. METHODS AND RESULTS: The study group comprised 149 patients (58 older and 91 younger) who received a final diagnosis of PE and were retrospectively evaluated. The severity of PE was assessed by calculating the pulmonary vascular obstruction scores (PVOs) scintigraphically: PVOs > or = 50% was defined as severe disease. Dyspnea was the most frequent symptom in both groups. Chest pain and hemoptysis were less frequent in older patients (48.3% vs 79.1%, p = 0.001; 6.9% vs 20.9%, p = 0.021, respectively) whereas syncope occurred more often in the older group (27.6% vs 9.9%, p = 0.005). PVOs > or = 50% occurred in 55.1% of older and 32.9% of younger patients (odds ratio: 1.67, 95%confidence interval: 1.118-2.507, p = 0.013). CONCLUSIONS: The clinical presentation of PE can be subtle or atypical in elderly patients and hence they may have more severe disease. Therefore, a high clinical suspicion is required in order to prevent delays in diagnostic work-up and initiation of appropriate treatment. PMID- 16041172 TI - Chronic beta-adrenergic receptor stimulation enhances the expression of G-Protein coupled receptor kinases, GRK2 and GRK5, in both the heart and peripheral lymphocytes. AB - BACKGROUND: Enhanced expression of G protein-coupled receptor kinase (GRK) has been reported in failing hearts and in the present study the stability of enhanced GRK mRNA expression, and the correlation between the expression level of GRK mRNA in peripheral lymphocytes and in the heart were both evaluated. METHODS AND RESULTS: Isoproterenol was injected into rats for 2 weeks, and then GRK5 mRNA was assessed by quantitative reverse transcriptase-palymerase chain reaction. An enhanced expression of cardiac GRK5 mRNA was observed even after 4 weeks of recovery. The isoproterenol-induced increased expression of GRK2 and GRK5 mRNA was equally observed in the heart and lymphocytes, and there was a close correlation between the heart and lymphocytes in the level of expression of each GRK mRNA. CONCLUSIONS: The GRK mRNA level is maintained at a high level for a long period without continuous beta-adrenergic receptor stimulation. The level in circulating lymphocytes could be used as a surrogate marker to estimate the level of cardiac GRK expression and, presumably, the beta-adrenergic receptor function of cardiomyocytes. PMID- 16041173 TI - Prediction of prognosis in the UM-X7.1 hamster model of congestive heart failure using the Tei index. AB - BACKGROUND: Cardiac function is difficult to evaluate in small animal models of heart disease. The Doppler Tei index is a simple and non-invasive measure that can express global cardiac function even in small animal models of congestive heart failure. However, its ability to predict prognosis has not been evaluated. METHODS AND RESULTS: We tested the hypothesis that cardiac functional indices, such as the Tei index, can predict the prognosis of hamsters with cardiac dysfunction. The Tei index, defined as the sum of the isovolume contraction and relaxation time divided by ejection time, and the percent fractional shortening of the left ventricle was measured in 48 anesthetized male hamsters (19.7+/-0.4 weeks old) with cardiac dysfunction (UM-X7.1), using Doppler and 2-dimensional echocardiography. The hamsters were separated into 2 groups based on the median Tei index (0.50) and % fractional shortening (FS) (21%). Kaplan-Meier analysis determined the survival rates of the groups. Both the Tei index and %FS enabled significant distinction of better and poorer survival (p < 0.01), and the survival curves were less overlapped when the animals were separated according to the Tei index. CONCLUSION: The Tei index can predict prognosis in a small animal model of heart failure. PMID- 16041174 TI - Genetic analysis in a patient with recurrent cardiac myxoma and endocrinopathy. AB - A 60 year-old male was referred for treatment of a cardiac myxoma in the right atrium. He had a past history of left atrial cardiac myxoma at age 49 and pituitary microadenoma related to acromegaly at age 55. He did not have a family history of cardiac neoplasm or endocrinopathy. The intracardiac tumor was resected and its pathology was compatible with myxoma. A diagnosis of Carney complex (CNC) was made because the diagnostic criteria of this neoplastic syndrome were satisfied by the presence of recurrent cardiac myxoma, endocrine tumor and spotty skin pigmentation. In genetic analysis novel frame shift mutation was detected in exon 2 in a heterozygous fashion in the causative gene of CNC, protein kinase A regulatory subunit 1 alpha (PRKAR1A). This genetic mutation is thought to cause haplo-insufficiency of PRKAR1A resulting in tumorigenesis. Although it is the most common, usually benign, cardiac tumor, myxoma can cause a critical clinical situation and thus detecting the PRKAR1A mutation can assist with prognosis. PMID- 16041175 TI - Successful surgical treatment of heart failure and ventricular tachycardia in a patient with arrhythmogenic right ventricular dysplasia with cardiomyopathy. AB - A 70-year-old male patient with arrhythmogenic right ventricular dysplasia/cardiomyopathy demonstrating frequent attacks of ventricular tachycardia (VT) as well as heart failure underwent surgical treatment. Although the patient had severe regurgitation at the mitral and tricuspid valves, the contractility of the right and left ventricles was almost maintained. Annuloplasty of both valves abolished the regurgitation and very effectively controlled heart failure. Surgical cryoablation was performed on the lesion showing the earliest potential before the ORS complex during VT and the arrhythmia was terminated. However, a cardioverter defibrillator was implanted to prevent new VT caused by disease progression. PMID- 16041176 TI - Acute myocardial infarction in a patient with essential thrombocythemia: successful treatment with percutaneous transluminal coronary recanalization. AB - A 65-year-old woman with essential thrombocythemia (ET) was admitted to hospital where she was diagnosed as acute myocardial infarction (AMI). Because of abundant thrombus of right coronary arteries, percutaneous transluminal coronary recanalization by administration of urokinase was selected as the reperfusion therapy, resulting in successful revascularization with Thrombolysis in Myocardial Infarction grade III coronary flow. The maximum creatine kinase reached 507 IU/L, and left ventriculography performed at 1 month after initiation of both anticoagulant and antiplatelet therapies revealed reduced motion in the inferior wall with an ejection fraction of 57%. Despite good recovery of left ventricular function, bleeding complications, such as epistaxis or ecchymoma, which did not require blood transfusion, occurred during the clinical course. Because ET causes not only thrombus formation but also bleeding tendency, it is very important to carefully follow-up any clotting abnormality in AMI patients with ET. PMID- 16041177 TI - Coexistence of the permanent form of junctional reciprocating tachycardia and atrial tachycardia. AB - This case report describes a patient with the permanent form of junctional reciprocating tachycardia coexisting with atrial tachycardia. A detailed electrophysiological study established the diagnosis, and radiofrequency catheter ablation abolished both arrythmias. PMID- 16041178 TI - Evaluation of hyperacute stroke using perfusion computed tomography. AB - Evaluation of the cerebral circulation dynamics in patients with stroke soon after onset is extremely important for planning the optimum treatment. Perfusion computed tomography (CT) was evaluated in 118 patients with stroke within 10 hours of onset in whom initial CT showed no ischemic change. The mean transit time (MTT), cerebral blood flow (CBF), and cerebral blood volume (CBV) perfusion CT maps were visually evaluated in three slices covering the body of the lateral ventricle, the basal ganglia, and the pons, and the ratios of the values in regions of interest (ROIs) in the ischemic lesion and the symmetrical location in the opposite hemisphere were calculated (ROI ratio of regional (r) MTT, rCBF, and rCBV). The location of occlusion was confirmed by angiography performed on the same day in 106 cases and the location of infarction by later magnetic resonance (MR) imaging. MTT maps correctly identified 44 of 46 cases of ischemia in the carotid system, 20 of 29 cases of ischemia in the vertebrobasilar system, and 11 of 35 cases of ischemia in perforator regions. Eight cases could not be identified by perfusion CT, angiography, or MR imaging. The ROI ratios at the upper boundary of infarction (n = 18) were: rCBF 0.574 +/- 0.220 (mean +/- SD) and rCBV 0.972 +/- 0.276, and at the lower boundary of non-infarction (n = 24) were: rCBF 0.504 +/- 0.247 and rCBV 0.815 +/- 0.169; showing a statistical significance of p = 0.348 for CBF and p = 0.026 for CBV (unpaired t-test). The perfusion CT MTT maps correlated well with the angiographical findings for the carotid system, but poorly for the vertebrobasilar system and the perforator regions. A rCBF ratio of 0.5 and rCBV ratio of 0.9 were established for the boundaries of ischemia. PMID- 16041179 TI - Efficacy of edaravone, a free radical scavenger, for the treatment of acute lacunar infarction. AB - The effect of edaravone as an inhibitor of ischemic brain damage in addition to routine treatment was retrospectively examined in 70 patients with lacunar infarction who were admitted within 24 hours of symptom onset. Clinical status was assessed using the National Institutes of Health Stroke Scale (NIHSS). The modified Rankin Scale (MRS) was used to assess clinical outcomes at 3 months after onset, with a good outcome defined as MRS score < or =2. Risk factors were also evaluated, including evidence of hypertension, diabetes mellitus, hyperlipidemia, coronary heart disease, and a history of smoking longer than 2 months. The probability of a good outcome and independence at 3 months was assessed by backward stepwise logistic regression analysis based on the maximum likelihood ratio. Administration of edaravone yielded an odds ratio with multivariate adjustment of 6.49 (95% confidence interval, 1.35 to 50.32; p < 0.05) for a good outcome at 3 months. Higher baseline NIHSS score and higher age also adversely affected the outcome at 3 months (p < 0.005). Administration of edaravone improves the outcome of patients with lacunar infarction. PMID- 16041180 TI - Optic nerve evoked potentials elicited by electrical stimulation. AB - This study investigated whether the optic nerve evoked potential (ONEP) elicited by electrical stimulation of the optic nerve can serve as a reliable intraoperative indicator of visual function. In the experimental study, two silver-ball stimulating electrodes were placed on the dog optic nerve adjacent to the apex of the orbit and one recording electrode was placed on the optic nerve near the chiasm. The nerve was stimulated with 0.1 to 10 mA rectangular pulses. Stable and reproducible ONEPs were obtained. The ONEPs were not influenced by electromyographic potentials and were recorded more clearly on the optic nerve than on the surrounding tissue. Stepwise incremental transection of the thickness of the nerve resulted in incremental amplitude reduction proportional to the transected area. No response was recorded after complete sectioning of the nerve. In the clinical study, recordings were obtained from 15 patients after craniotomy to treat parasellar tumors or cerebral aneurysms. Reproducible ONEPs were recorded intraoperatively from the electrode placed on the optic nerve near the chiasm in 14 of 15 patients. In the remaining patient, the ONEP, recorded only after tumor removal because the optic nerve was stretched and extremely thin, was remarkably small and the patient developed unilateral blindness postoperatively. These experimental and clinical results suggest the possibility of intraoperative monitoring of visual function in patients undergoing craniotomy for the treatment of lesions near the optic nerve. PMID- 16041181 TI - Surgical clipping of a recurrent small saccular aneurysm after repeated coil embolization. AB - A 59-year-old healthy woman presented with sudden onset of severe headache. Computed tomography and digital subtraction angiography (DSA) demonstrated subarachnoid hemorrhage (grade I according to the Hunt and Hess classification) due to a ruptured small right posterior cerebral artery (PCA) aneurysm. The ruptured PCA aneurysm was completely embolized with three Guglielmi detachable coils (GDCs). However, follow-up DSA 3 months after the initial coiling confirmed refilling of the aneurysm. The aneurysm was successfully re-embolized with two GDCs. Follow-up DSA 10 months later revealed regrowth of the aneurysm. Surgical clipping was performed without compromising the parent vessels. Long-term angiographic follow up is necessary even in patients with small saccular aneurysms which are apparently completely embolized by endovascular coil treatment. PMID- 16041182 TI - Endovascular treatment of vertebral artery aneurysm manifesting as progressive hemifacial spasm. AB - A 62-year-old woman presented with right hemifacial spasm persisting for 6 months. Brain magnetic resonance imaging and digital subtraction angiography showed a wide-neck aneurysm of the intracranial portion of the right vertebral artery. The patient underwent endovascular trapping of the aneurysm by coil embolization of the parent vessel on both sides of the aneurysm. The patient experienced gradual disappearance of the hemifacial spasm within 3 months. No relapses occurred during a follow-up period of 3 years. Magnetic resonance imaging revealed shrinkage of the vertebral artery aneurysm which had compressed the facial nerve. Endovascular trapping of a vertebral artery aneurysm can be used to treat hemifacial spasm caused by an aneurysm instead of surgical microvascular decompression. PMID- 16041183 TI - Postoperative hyperperfusion associated with steal phenomenon caused by a small arteriovenous malformation. AB - A 41-year-old woman presented with a small occipital arteriovenous malformation (AVM) manifesting as headache. Cerebral angiography showed an AVM in the right occipital lobe fed by the right temporooccipital artery and draining into the superior sagittal sinus and right transverse sinus. Single photon emission computed tomography showed the steal phenomenon in the ipsilateral temporal cortex fed by the main feeding artery preoperatively, and hyperperfusion in the same cortex after removal of the AVM. Postoperative systolic blood pressure was maintained between 100 and 120 mmHg to avoid disastrous hemorrhagic complications. Cerebral blood flow evaluation before and after surgery is important to avoid postoperative disastrous complications even in patients with small AVM. PMID- 16041184 TI - Skull metastasis from clear cell chondrosarcoma. AB - A 67-year-old man presented with left lower cranial nerve paresis and dysfunction of the left cerebellar hemisphere 4 years after amputation of the left lower leg because of clear cell chondrosarcoma (CCC). Neuroimaging studies showed an osteolytic extradural mass with homogeneous enhancement in the left posterior fossa. Bone scintigraphy disclosed a single high-uptake lesion at the same site. The tumor was removed totally via a left suboccipital craniotomy. Histological examination found mainly clear cells arranged in a microlobular pattern separated by thin fibrovascular stroma. The nuclei were regular with few mitotic figures. Immunohistochemical staining showed the tumor cells reacted intensely for both S 100 protein and vimentin. Osteoclast-like multinucleated giant cells were found at the periphery of the lobules. The primary tumor showed the same findings and the metastatic tumor manifested no malignant change. The histological diagnosis was metastatic CCC. CCC is a very rare neoplasm with slow growth and low-grade malignancy. Distant metastasis is rare but can occur in the skull base bone despite radical resection of the primary tumor. Osteolytic findings of homogeneous enhancement on magnetic resonance imaging and a high uptake on bone scintigraphy are indicative of metastatic tumor from previous CCC. PMID- 16041185 TI - Solitary fibrous tumor of the thoracic spinal cord. AB - A 63-year-old woman presented with a rare case of primary solitary fibrous tumor (SFT) occurring in the extramedullary thoracic spinal cord. T1-weighted magnetic resonance (MR) imaging showed the tumor as a mildly hypointense area with homogeneous enhancement by gadolinium. T2-weighted MR imaging showed a hypointense mass with peritumoral edema. The tumor arose from one of the posterior spinal roots, with no attachment to the dura. The tumor was clearly circumscribed from the surrounding cord tissue and easily removed. Histological examination showed the tumor predominantly consisted of spindle cells separated by abundant collagen matrix fibers. Tumor cells were strongly positive for vimentin and CD34, but negative for glial fibrillary acidic protein, S-100 protein, epithelial membrane antigen, myelin basic protein, and keratin. SFT should be considered in the differential diagnosis of spindle cell central nervous system neoplasms, although SFT is extremely rare in the spinal cord. PMID- 16041186 TI - Mature spinal teratoma associated with thickened filum terminale. AB - A 30-year-old man presented with an intradural spinal teratoma with thickened filum terminale manifesting as urinary and sexual disturbances, and low back pain persisting for 4 years. Spinal magnetic resonance imaging revealed thickened filum terminale containing a heterogeneously enhanced intradural lesion extending from the L-3 to L-4 levels and in contact with the conus medullaris. The filum terminale was incised and the tumor was totally resected. The histological diagnosis was mature teratoma consisting of three germ cell layers. The patient's complaints had completely resolved 6 months later. PMID- 16041187 TI - Progression of a lumbar spinal osteoblastoma. AB - A 24-year-old woman presented with a lumbar spinal osteoblastoma manifesting as a 5-year history of low back pain radiating to the left foot. Neuroimaging showed suspicious hypertrophy of the left L4-5 facet which transformed in 3 years to an expansile mass lesion that compressed the dura mater and neural structures. Primary benign bone tumors such as osteoblastoma and osteoid osteoma should be considered in the differential diagnosis of back pain and the patients should be followed up carefully. PMID- 16041188 TI - Supplementary comment on "experimental model of intracranial arteriovenous shunting in the acute stage". PMID- 16041189 TI - Intellectual property. PMID- 16041190 TI - The erythrosense as a real-time biomarker to reveal the presence of enhanced red blood cell aggregability in atherothrombosis. AB - Both anemia and inflammation might be present in individuals with atherothrombosis. We have evaluated the eventual influence of these 2 variables on the degree of erythrocyte adhesiveness/aggregation in the peripheral blood of 583 women and 402 men with various atherothrombotic risk factors and vascular events. It turned out that both anemia and inflammation (highly sensitive C reactive protein concentrations) influence the degree of cell adhesiveness/aggregation and that there is no interaction between them. Thus, the degree of erythrocyte adhesiveness/aggregation might have the diagnostic advantage of being enhanced in individuals with atherothrombosis who have inflammation and no anemia as well as those who have anemia and no inflammation. These findings might help to turn a phenomenon of hemorheological relevance into a diagnostic tool for the detection of individuals at risk of an acute ischemic event. PMID- 16041191 TI - Efficacy of unfractionated heparin for thromboembolism prophylaxis in medical patients. AB - A common mode of deep vein thrombosis prophylaxis in medical inpatients is unfractionated heparin 5000 U subcutaneously (s.q.) twice daily. We examined the evidence in favor of using this dose of heparin in this group of patients. MEDLINE was searched for studies using the words deep vein thrombosis prophylaxis and heparin. All randomized controlled trials comparing heparin and placebo or heparin and a low molecular weight heparin were used. Relative risk was 0.4 (95% confidence interval 0.22-0.73) in studies comparing heparin 5000 U s.q. b.i.d. with placebo. Relative risk was 0.28 (95% confidence interval 0.21-0.38) in studies comparing heparin 5000 units s.q. t.i.d. versus placebo. In studies comparing unfractionated heparin with enoxaparin relative risk was 1.42 (95% confidence interval 0.99-2.05). Heparin 5000 U s.q. b.i.d. is less efficacious than low molecular weight heparins and unfractionated heparin 5000 U s.q. t.i.d. PMID- 16041192 TI - The processing of the selective M1 agonist CDD-0102-J by human hepatic drug metabolizing enzymes. AB - The in vitro metabolism of the selective M1 muscarinic agonist CDD-0102-J was evaluated in heterologous systems expressing individual human cytochrome P-450 (CYP) isoenzymes and also in suspensions of cryopreserved human hepatocytes. In all experiments, the metabolism of CDD-0102-J was characterized based on its rate of disappearance using an HPLC assay since no metabolites have as yet been characterized. The human CYP isoenzymes used were CYP1A2, 2A6, 2B6, 2C8, 2C19, 2D6, and 3A4. Measurable decreases in CDD-0102-J concentrations over time were detectable only in systems containing either CYP2D6 or CYP2C8, although the unbound in vitro clearance was more than 20 times larger for CYP2D6 (7.6 mL h(-1) nmol(-1)) than for CYP2C8 (0.35 mL h(-1) nmol(-1)). When scaled to in vivo hepatic clearance based on just CYP2D6 and CYP2C8, the projected hepatic clearance for CDD-0102-J was 7.7 L h(-1), which corresponded closely with the hepatic clearance of 8.4 L h(-1) scaled from experiments using cryopreserved human hepatocytes. PMID- 16041193 TI - Colesevelam HCl and ezetimibe combination therapy provides effective lipid lowering in difficult-to-treat patients with hypercholesterolemia. AB - This study investigated the effects of colesevelam hydrochloride (WelChol; Sankyo Pharma, Parsippany, NJ) and ezetimibe (Zetia; Merck/Schering Plough Pharmaceuticals, North Wales, PA), alone and in combination, for the treatment of hypercholesterolemia in patients who were intolerant to, or refused, HMG-Co-A reductase inhibitor (statin) therapy. Combination therapy with colesevelam HCl/ezetimibe resulted in an additional reduction in low-density lipoprotein cholesterol (LDL-C) and non-high-density lipoprotein cholesterol (HDL-C) levels of approximately 20% (P < 0.005) and 16% (P < 0.01), respectively, compared with monotherapy with either agent. Total cholesterol, LDL-C, and non-HDL-C levels were within National Cholesterol Education Program Adult Treatment Panel III target ranges at the end of the combination therapy regimen in 10 of 12 patients. In conclusion, colesevelam HCl/ezetimibe combination therapy appears to be an efficacious and well-tolerated alternative for patients with hypercholesterolemia. PMID- 16041194 TI - Lidocaine patch treatment in patients with low back pain: results of an open label, nonrandomized pilot study. AB - This prospective, 6-week, multicenter, open-label, nonrandomized pilot study was designed to assess the effectiveness and safety of a lidocaine patch 5% in patients with low back pain (LBP). Patients with moderate to severe LBP, defined as acute/subacute (< 3 months, n = 21), short-term chronic (3-12 months, n = 33), or long-term chronic (> 12 months, n = 77), were recruited from 5 clinics; participants applied < or = 4 patches (560 cm total) once daily to area of maximal LBP as add-on treatment through week 2, with the option to taper concomitant analgesics during weeks 3-6. Scores on Brief Pain Inventory (BPI) were obtained at weeks 2 and 6. Safety analyses included reports of adverse events (AEs) and skin sensitivity to pinprick/light touch. Significant improvements in average daily pain intensity on the BPI were noted at weeks 2 and 6 (P < or = 0.001). Significant improvements in pain interference with quality of life (QOL) were noted for all BPI measures of QOL at weeks 2 and 6 for the acute/subacute (P < or = 0.007) and long-term chronic LBP groups (P < 0.0001) and for 5 of 7 BPI measures for the short-term chronic LBP group (P < or = 0.042). Fifty-eight percent of patients reported being "satisfied" or "very satisfied" with treatment. The lidocaine patch was well tolerated. Most common AEs were dizziness and rash (n = 5, 3.8%), and most AEs (80%) were mild to moderate in intensity. Significant improvement in pain intensity and QOL in this cohort of LBP patients was noted during treatment with the lidocaine patch 5%. Controlled clinical trials are warranted. PMID- 16041195 TI - Application of two-point assay of digoxin serum concentration in studying population pharmacokinetics in Egyptian pediatric patients with heart failure: does it make sense? AB - Digoxin pharmacokinetics (PK) was studied among a selected group of Egyptian pediatric patients (n = 40) with an age range of 0.33 to 15 years. All the patients had heart failure and were maintained on i.v. digoxin (10 microg/kg/d in 2 equal doses). For population PK analysis, 2 serum samples of digoxin were taken per patient. From 30 patients' trough (before the next dose) and 4 hours postdose samples were obtained, while in the other 10 patients, 0.5- and 6-hour postdose samples were taken. Serum concentrations were measured by fluorescence polarization immunoassay. PK modeling was performed using NONMEM software on log transformed serum digoxin data. The best structural covariate-free model was a linear 2-compartment model with an exponential error model for intersubject variability and an additive model for intrasubject variability. Serum creatinine (SCR) was a significant covariate for clearance. The final population PK parameters were CL (L/h) = 0.388 - [0.78 x (SCR-0.6)], V1 (L/kg) = 1.38, Q (L/h/kg) = 0.48, V2 (L/kg) = 9.11, where CL is the total body clearance, V1 and V2 are the apparent volumes of distribution in the central and peripheral compartments, and Q is intercompartment clearance. A bootstrap resampling for internal validation achieved excellent agreement with the original data sets for PK parameters. In conclusion, 2 points of digoxin concentration allow good regression analysis for clearance-covariate relationship. The inclusion of SCR into the final model might allow better selection of initial maintenance dose of the drug. A prospective study on larger sample size of pediatric patients is recommended for clinical validation of the final model. PMID- 16041196 TI - A mechanism for the potential proarrhythmic effect of acidosis, bradycardia, and hypokalemia on the blockade of human ether-a-go-go-related gene (HERG) channels. AB - Many drugs are proarrhythmic by inhibiting the cardiac rapid delayed rectifier potassium channel (IKr). In this study, we use quinidine as an example of highly proarrhythmic agent to investigate the risk factors that may facilitate the proarrhythmic effects of drugs. We studied the influence of pacing, extracellular potassium, and pH on quinidine's IKr blocking effect, all potential factors influencing quinidine's cardiac toxicity. Since the HERG gene encodes IKr, we studied quinidine's effect on HERG expressed in Xenopus oocytes by the 2 electrode voltage clamp technique. When extracellular K+ was 5 mmol/L, quinidine blocked the HERG current dose dependently, with an IC50 of 6.3 +/- 0.2 micromol/L. The blockade was much more prominent at more positive membrane potentials. The inhibition of HERG by quinidine was not use dependent. There was no significant difference between block with or without pacing. When extracellular K+ was lowered to 2.5 mmol/L, the current inhibition by quinidine was enhanced, and IC50 decreased to 4.6 +/- 0.5 micromol/L. At 10 mmol/L extracellular K+, there was less inhibition by quinidine and the IC50 was 11.2 +/ 3.1 micromol/L. Extracellular acidification decreased both steady state and tail currents of HERG. We conclude that the inhibitory effect of quinidine on IKr was decreased with extracellular acidification, which may produce heterogeneity in the repolarization between normal and ischemic cardiac tissue. Thus, the use independent blockade of IKr by QT-prolonging agents such as quinidine may contribute to cardiac toxicity with bradycardia, hypokalemia, and acidosis further exaggerating the proarrhythmic potential of these agents. PMID- 16041197 TI - Incremental cost savings 6 months following initiation of insulin glargine in a Medicaid fee-for-service sample. AB - This study assessed the role of insulin glargine use on the short-term costs of diabetes care from a state Medicaid fee-for-service reimbursement perspective. A retrospective claims analysis was performed for 20% of Medicaid recipients in California (Medi-Cal) between November 2000 and September 2002. Each patient with continuous enrollment at least 6 months before and after the insulin glargine index date was matched with 2 reference patients by age, gender, diabetes diagnosis, and enrollment dates. Costs were calculated for emergency department, inpatient, outpatient, and pharmacy claims paid, and changes from baseline were determined. Incremental cost savings were estimated from baseline cost in the insulin glargine group multiplied by the difference in percentage of changes from baseline. Of the 1018 insulin glargine users, 267 satisfied inclusion criteria and were matched to 534 reference patients. Baseline treatment costs were 2.7 fold higher in the insulin glargine group. Absolute cost reductions in the insulin glargine and reference groups were 185 dollars and 72 dollars per patient, respectively. Incremental cost savings were 69 dollars per insulin glargine user, including more than 200 dollars in incremental savings for inpatient claims. Hypoglycemia decreased from 9.5% to 3.8% of inpatient claims in the insulin glargine group but remained 1.1% throughout in the reference group. Insulin glargine use was associated with reduced inpatient hypoglycemia-related claims paid and short-term reductions in the inpatient and total costs of diabetes-related care. Additional studies are warranted to assess the influence of insulin glargine on diabetes-related costs. PMID- 16041198 TI - Management of peritonsillar abscess: needle aspiration versus incision and drainage versus tonsillectomy. AB - Peritonsillar abscess is the most common deep infection of the head and neck. This article focuses on the different methods to manage this infection. Efficacy, cost-effectiveness, patient discomfort, recovery time, and possibility of recurrence are the issues considered to determine the best treatment option. PMID- 16041199 TI - Cluster headache pharmacotherapy. AB - Cluster headache is a well-known primary headache syndrome with a prevalence of about 5/10,000 of the adult population, making it much less common than migraine. Diagnostic terms such as histaminic cephalalgia, Horton's headache and ciliary neuralgia have been used for what is now known as cluster headache. This disorder can be differentiated from migraine by clinical and pathophysiologic features. Cluster headache also exhibits a differing therapeutic response to medications when compared with migraine. The pharmacologic treatment of cluster is reviewed in this article. In contrast to migraine, men are 3-4 times more likely to be diagnosed with cluster headache than women, and the cluster headache population is older. Cluster attacks are known for their brief intense unilateral excruciating pain during susceptible periods known as cluster periods, which typically last weeks. Attack-free months generally follow. Pain is experienced in the distribution of the trigeminal nerve, with unilateral autonomic features. Most patients are successfully managed with medical therapy. Medication management can be divided into abortive treatments for an ongoing attack and prophylactic treatment. Prophylaxis aims to induce and maintain a remission. There are a variety of different medications for abortive and prophylactic therapy, accompanied by a variable amount of evidence-based medicine. For patients refractory to medical management, interventional procedures are available as a last resort. Most procedures are directed against the sensory trigeminal nerve and associated ganglia, eg, anesthetizing the sphenopalatine ganglion. PMID- 16041200 TI - Hypertension and cognitive functioning among the elderly: an overview. AB - This paper reviews the longitudinal studies in which the association between hypertension and cognitive functioning has been explored. The findings of these studies are far from being consistent: positive, adverse, or no relationships between cognitive performance and blood pressure have been reported. In some studies, cognitive functioning was related only to systolic blood pressure, whereas in others, it was related to diastolic blood pressure alone. Most probably, the inconsistency is due to the variability in research design, the measures employed, and data management. Some of the main methodological issues are the wide range in the age of the subjects and the variability in hypertension at baseline. Length of exposure to hypertension was rarely considered, and the search for the possible effects of the different antihypertensive treatments on cognitive ability is in its infancy. Many studies seek to identify cognitively impaired elderly and do not distinguish between the different types of dementia or the different cognitive domains. Thus, the possibility that hypertension affects specific cognitive functions has not been explored. The use of subdivisions of blood pressure values may have obscured nonlinear relationship. Nevertheless, the pattern that emerges is that midlife hypertension enhances cognitive impairment later in life, but at old age, mild hypertension may increase cognitive performance. Directions for future research to further elucidate the complex relationship between age, blood pressure, and cognition are discussed. PMID- 16041201 TI - Use of nebulized budesonide in two critical patients with hydrocarbon intoxication. AB - Reports of acute naphtha (a petroleum distillate) intoxication, particularly those containing treatment and toxicological data, are poorly verified in the literature. Systemic steroids have not usually been advised for routine management, but no previous studies have been reported on the effects of corticosteroid inhalations. In this study, two critical pediatric patients with chemical pneumonia due to naphtha ingestion have been reported who were effectively managed with nebulized budesonide. PMID- 16041202 TI - Primary aldosteronism with HIV infection: important considerations when using the aldosterone:renin ratio to screen this unique population. AB - Primary aldosteronism (PA) was initially thought to be a rare cause of hypertension. Currently, the screening modality for PA in patients with presumed secondary hypertension is the aldosterone:renin ratio (ARR). Since the advent of the ARR, the prevalence rates of PA have increased dramatically in the general population. Recently however, several reviews have suggested flaws in the ARR, leading to a possible overestimation of the prevalence of this disease. Within the HIV-positive population, there are few documented cases of PA however. With the increasing reliance on the ARR to screen and diagnose this disorder, this number is likely to rise. It is now known that the HIV has rennin-like properties. Therefore, HIV infection itself can inappropriately alter the ARR. Additionally, the medication used to treat HIV infection can also affect this value. Although the ARR is a convenient tool for screening patients with possible PA, there are multiple associated concerns particularly when it is applied to the HIV-positive population. Therefore, further testing is essential in all HIV positive patients in whom PA is suspected. PMID- 16041203 TI - Racial disparity in surgical complications in New York State. AB - OBJECTIVE: To examine the relationship between race and surgical complications. SUMMARY BACKGROUND DATA: Blacks have been reported to experience higher rates of surgical complications than whites, but the reasons are not known. METHODS: The effect of the black race on risk of any surgical complication (from the Agency for Healthcare Research and Quality's patient safety indicators) was examined using New York State (NYS) hospital discharge data from 1998 to 2000. Sequential, hierarchical analyses controlled for: 1) patient age and gender, 2) morbidity length of stay, 3) individual social factors, 4) hospital characteristics, and 5) ecologic factors (region of state, percent black and Medicaid annual discharges, and mean income of admitted patients). RESULTS: Following adjustment for patient age and gender, blacks had 65% higher odds for a surgical complication. Further adjustment for comorbidity and length of stay (LOS) reduced the odds substantially to 1.18. Additional adjustment for American Hospital Association hospital characteristics essentially eliminated the risk. Final adjustment for hospital ecologic variables reduced the odds to 1.0. CONCLUSIONS: Higher rates of surgical complications among blacks than whites in NYS are primarily explained by differences in comorbidity LOS and the hospital where the surgery occurred. PMID- 16041204 TI - The long arm of health care disparity. PMID- 16041205 TI - Local recurrence after hepatic radiofrequency coagulation: multivariate meta analysis and review of contributing factors. AB - OBJECTIVE: The purpose of this study was to analyze the factors that influence local recurrence after radiofrequency coagulation of liver tumors. SUMMARY BACKGROUND DATA: Local recurrence rate varies widely between 2% and 60%. Apart from tumor size as an important risk factor for local recurrence, little is known about the impact of other factors. METHODS: An exhaustive literature search was carried out for the period from January 1, 1990 to January 1, 2004. Only series with a minimal follow-up of 6 months and/or mean follow-up of 12 months were included. Univariate and multivariate meta-analyses were carried out. RESULTS: Ninety-five independent series were included, allowing the analysis of the local recurrence rate of 5224 treated liver tumors. In a univariate analysis, tumor dependent factors with significantly less local recurrences were: smaller size, neuroendocrine metastases, nonsubcapsular location, and location away from large vessels. Physician-dependent favorable factors were: surgical (open or laparoscopic) approach, vascular occlusion, general anesthesia, a 1-cm intentional margin, and a greater physician experience. In a multivariate analysis, significantly less local recurrences were observed for small size (P < 0.001) and a surgical (versus percutaneous) approach (P < 0.001). CONCLUSIONS: Radiofrequency coagulation by laparoscopy or laparotomy results in superior local control, independent of tumor size. The percutaneous route should mainly be reserved for patients who cannot tolerate a laparoscopy or laparotomy. The short term benefits of less invasiveness for the percutaneous route do not outweigh the longer-term higher risk of local recurrence. PMID- 16041206 TI - Two-surgeon technique for hepatic parenchymal transection of the noncirrhotic liver using saline-linked cautery and ultrasonic dissection. AB - OBJECTIVE: The purpose of this study was to analyze our experience with saline linked cautery in hepatic surgery. SUMMARY BACKGROUND DATA: Safe and efficient hepatic parenchymal transection is predicated on the ability to simultaneously address 2 tasks: parenchymal dissection and hemostasis. To date, no single instrument has been designed that addresses both of these tasks. Saline-linked cautery is now widely used in liver surgery and is reported to decrease blood loss during liver transection, but data on its exact benefits are lacking. METHODS: From a single institution, prospective liver surgery database, we identified 32 consecutive patients with noncirrhotic livers who underwent resection for primary or metastatic disease using a 2-surgeon technique with saline-linked cautery and ultrasonic dissection (SLC+UD) from December 2002 to January 2004. From the same database, we identified a contemporary and matched set of 32 patients who underwent liver resection with similar indications using ultrasonic dissection alone (UD alone). Operative and anesthetic variables were retrospectively analyzed to identify differences between the 2 groups. RESULTS: The 2 groups were equivalent in terms of age, gender, tumor histology, tumor number, and tumor size. The UD+SLC group had a decreased duration of inflow occlusion (20 minutes versus 30 minutes, P = 0.01), blood loss (150 mL versus 250 mL, P = 0.034), and operative time (187 minutes versus 211 minutes, P = 0.027). Postoperative liver function and complication rates were similar in each group. CONCLUSIONS: The 2-surgeon technique for liver parenchymal transection using SLC and UD in noncirrhotic livers is safe and may provide advantages over other techniques. PMID- 16041207 TI - Increase of survival benefit in advanced resectable colon cancer by extent of adjuvant treatment: results of a randomized trial comparing modulation of 5-FU + levamisole with folinic acid or with interferon-alpha. AB - BACKGROUND: The benefit of adjuvant therapy in curatively resected lymph node positive colon cancer was established using 5-fluorouracil (5-FU) and levamisole (LEV) for 12 months. 5-FU cytotoxicity can be modulated by folinic acid (FA) or interferon-alpha (INF-alpha). The aim of this study was to investigate the efficacy of modulating 5-FU+ LEV by either FA or IFN-alpha in the adjuvant treatment of high-risk colon cancer. METHODS: Patients with curatively resected colon cancer (stages UICC IIb and III) were stratified according to T, N, and participating center and randomized to receive a 12-month treatment using 5-FU + LEV alone or in combination with FA or IFN-alpha. RESULTS: A total of 855 of 904 entered patients (94.6%) were eligible. The median follow-up of all eligible patients was 4.6 years. Addition of FA to 5-FU + LEV improved recurrence-free and overall survival in comparison with 5-FU + LEV alone (P = 0.007 and P = 0.004, respectively, 1-sided). The 5-year overall survival rates were 60.5% (95% confidence interval, 54.3-66.7) and 72.0% (95% confidence interval, 66.5-77.5) for 5-FU + LEV and 5-FU + LEV + FA, respectively. Addition of INF-alpha showed a tendency to improve recurrence-free survival, however, without altering overall survival. Toxicities (WHO III + IV) were generally tolerable except one toxic death in the control arm and were observed in 9.9% of the patients receiving 5-FU + LEV alone and in 13.3% and in 30.7% of patients receiving additional FA and IFN alpha, respectively. CONCLUSIONS: Addition of IFN-alpha was associated with increased toxicity without markedly influencing the outcome and should therefore not be recommended for adjuvant treatment. Addition of FA increased the 5-year recurrence-free and overall survival rate by 9.3 and 11.5 percentage points, respectively. 5-FU + LEV + FA for 12 months may be an effective adjuvant treatment option for locally advanced high-risk colon cancer. PMID- 16041208 TI - Postoperative ERCP versus laparoscopic choledochotomy for clearance of selected bile duct calculi: a randomized trial. AB - OBJECTIVE: Prospectively evaluate whether for patients having laparoscopic cholecystectomy with failed trans-cystic duct clearance of bile duct (BD) stones they should have laparoscopic choledochotomy or postoperative endoscopic retrograde cholangiography (ERCP). SUMMARY BACKGROUND DATA: Clinical management of BD stones found at laparoscopic cholecystectomy in the last decade has focused on pre-cholecystectomy detection with ERCP clearance in those with suspected stones. This clinical algorithm successfully clears the stones in most patients, but no stones are found in 20% to 60% of patients and rare unpredictably severe ERCP morbidity can result in this group. Our initial experience of 300 consecutive patients with fluoroscopic cholangiography and intraoperative clearance demonstrated that, for the pattern of stone disease we see, 66% of patients' BD stones can be cleared via the cystic duct with dramatic reduction in morbidity compared to the 33% requiring choledochotomy or ERCP. Given the limitations of the preoperative approach to BD stone clearance, this trial was designed to explore the limitations, for patients failing laparoscopic trans cystic clearance, of laparoscopic choledochotomy or postoperative ERCP. METHODS: Across 7 metropolitan hospitals after failed trans-cystic duct clearance, patients were intraoperatively randomized to have either laparoscopic choledochotomy or postoperative ERCP. Exclusion criteria were: ERCP prior to referral for cholecystectomy, severe cholangitis or pancreatitis requiring immediate ERCP drainage, common BD diameter of less than 7 mm diameter, or if bilio-enteric drainage was required in addition to stone clearance. Drain decompression of the cleared BD was used in the presence of cholangitis, an edematous ampulla due to instrumentation or stone impaction and technical difficulties from local inflammation and fibrosis. The ERCP occurred prior to discharge from hospital. Mechanical and extracorporeal shockwave lithotripsy was available. Sphincter balloon dilation as an alternative to sphincterotomy to allow stone extraction was not used. Major endpoints for the trial were operative time, morbidity, retained stone rate, reoperation rate, and hospital stay. RESULTS: From June 1998 to February 2003, 372 patients with BD stones had successful trans-cystic duct clearance of stones in 286, leaving 86 patients randomized into the trial. Total operative time was 10.9 minutes longer in the choledochotomy group (158.8 minutes), with slightly shorter hospital stay 6.4 days versus 7.7 days. Bile leak occurred in 14.6% of those having choledochotomy with similar rates of pancreatitis (7.3% versus 8.8%), retained stones (2.4% versus 4.4%), reoperation (7.3% versus 6.6%), and overall morbidity (17% versus 13%). CONCLUSIONS: These data suggest that the majority of secondary BD stones can be diagnosed at the time of cholecystectomy and cleared trans-cystically, with those failing having either choledochotomy or postoperative ERCP. However, because of the small trial size, a significant chance exists that small differences in outcome may exist. We would avoid choledochotomy in ducts less than 7 mm measured at the time of operative cholangiogram and severely inflamed friable tissues leading to a difficult dissection. We would advocate choledochotomy as a good choice for patients after Billroth 11 gastrectomy, failed ERCP access, or where long delays would occur for patient transfer to other locations for the ERCP. PMID- 16041209 TI - Comparison of antimicrobial impregnation with tunneling of long-term central venous catheters: a randomized controlled trial. AB - OBJECTIVE: We sought to compare the impact of antimicrobial impregnation to that of tunneling of long-term central venous catheters on the rates of catheter colonization and catheter-related bloodstream infection. SUMMARY BACKGROUND DATA: Tunneling of catheters constitutes a standard of care for preventing infections associated with long-term vascular access. Although antimicrobial coating of short-term central venous catheters has been demonstrated to protect against catheter-related bloodstream infection, the applicability of this preventive approach to long-term vascular access has not been established. METHODS: A prospective, randomized clinical trial in 7 university-affiliated hospitals of adult patients who required a vascular access for > or = 2 weeks. Patients were randomized to receive a silicone central venous catheter that was either impregnated with minocycline and rifampin or tunneled. The occurrence of catheter colonization and catheter-related bloodstream infection was determined. RESULTS: Of a total of 351 inserted catheters, 346 (186 antimicrobial-impregnated and 160 tunneled) were analyzed for catheter-related bloodstream infection. Clinical characteristics were comparable in the 2 study groups, but the antimicrobial impregnated catheters remained in place for a shorter period of time (mean, 30.2 versus 43.8 days). Antimicrobial-impregnated catheters were as likely to be colonized as tunneled catheters (7.9 versus 6.3 per 1000 catheter-days). Bloodstream infection was 4 times less likely to originate from antimicrobial impregnated than from tunneled catheters (0.36 versus 1.43 per 1000 catheter days). CONCLUSIONS: Antimicrobial impregnation of long-term central venous catheters may help obviate the need for tunneling of catheters. PMID- 16041210 TI - Association of adult celiac disease with surgical abdominal pain: a case-control study in patients referred to secondary care. AB - BACKGROUND: Acute abdominal pain is the most common indication for surgical admission. Nonspecific abdominal pain (NSAP) may account for up to 40% of cases. There has been no published prospective study in which adult patients presenting with acute abdominal pain are investigated for celiac disease. AIMS: We aimed to assess the association of celiac disease with surgical abdominal pain. PATIENTS AND METHODS: A case-control study was undertaken involving 300 consecutive new unselected patients presenting with acute abdominal pain (in a university hospital) and healthy controls (age and sex matched) without abdominal pain (n = 300). Initial investigations for celiac disease were immunoglobulins, IgA/IgG anti-gliadin (AGA), and endomysial antibodies (EMA). Any patient with a positive IgA AGA, EMA, or only IgG AGA in the presence of IgA deficiency was offered a small bowel biopsy to confirm the diagnosis. RESULTS: : There were 33 patients with abdominal pain who had positive antibodies, of whom 9 had histologically confirmed celiac disease (6 EMA positive; 3 EMA negative). One antibody positive patient (EMA in isolation) declined duodenal biopsy and the remaining 23 had normal duodenal mucosa. Within the control group, there were 2 cases of celiac disease. Compared with matched controls the association of acute abdominal pain with celiac disease gave an odds ratio 4.6. (P = 0.068, 95% confidence interval, 1.11-19.05). When only considering NSAP the prevalence of celiac disease was highly significant at 10.5% (9 of 86, P = 0.006). Patients' symptoms improved on a gluten-free diet at 12- to 18-month follow-up. CONCLUSION: Celiac disease was diagnosed in 3% of patients who presented with unselected acute abdominal pain to secondary care. Targeting patients who have NSAP or celiac associated symptoms/diseases may improve the diagnostic yield. PMID- 16041211 TI - Internal anal sphincter function following lateral internal sphincterotomy for anal fissure: a long-term manometric study. AB - BACKGROUND: Anal fissure is a common and painful disorder. Its relation to hypertonic anal sphincter is controversial. The most common surgical treatment of chronic anal fissure is lateral internal sphincterotomy. OBJECTIVE: The aim of this study was to evaluate long-term manometric results of sphincter healing following lateral internal sphincterotomy. PATIENTS AND METHODS: Between 2000 and 2003, 50 patients with anal fissure were included in this study and underwent sphincterotomy; 12 healthy patients served as controls. All patients with anal fissure underwent manometric evaluation using a 6-channel perfusion catheter. All patients were examined 1 month before surgery and 1, 3, 6, and 12 months following surgery. The control group had 3 manometric evaluations 6 months apart. RESULTS: The mean basal resting pressure before surgery was 138 +/- 28 mm Hg. One month after surgery, the pressure dropped to 86 +/- 15 mm Hg (P < 0.0001) and gradually rose to a plateau at 12 months (110 +/- 18 mm Hg, P < 0.0001). At 12 months, the manometric pressure was significantly lower than the baseline (P < 0.0001). However, manometric measurements in the fissure group were still significantly higher than in the control group (110 +/- 18 versus 73 +/- 4.8 mm Hg, P < 0.0001). All patients were free of symptoms at the 12-month follow-up. CONCLUSION: Lateral internal sphincterotomy caused a significant decline in the resting anal pressure. During the first year following surgery, the tone of the internal anal sphincter gradually increased, indicating recovery, but still remained significantly lower than before surgery. However, postoperative resting pressures were higher than those in the control, and no patient suffered any permanent problems with incontinence, so this decrease may not be clinically significant. PMID- 16041212 TI - Prevalence of male and female sexual dysfunction is high following surgery for rectal cancer. AB - OBJECTIVE: To measure sexual function and quality of life (QOL) after rectal cancer treatment. SUMMARY BACKGROUND DATA: Previous studies on sexual function after rectal cancer treatment have focused on males and have not used validated instruments. METHODS: Patients undergoing curative rectal cancer surgery from 1980 to 2003 were administered a questionnaire, including the Female Sexual Function Index (FSFI) or International Index of Erectile Function (IIEF), and the EORTC QLQ-C30/CR-38. Multiple logistic regression was used to test associations of clinical factors with outcomes. RESULTS: Eighty-one women (81.0%) and 99 men (80.5%) returned the questionnaire; 32% of women and 50% of men are sexually active, compared with 61% and 91% preoperatively (P < 0.04); 29% of women and 45% of men reported that "surgery made their sexual lives worse." Mean (SD) FSFI and IIEF scores were 17.5 (11.9) and 29.3 (22.8). Specific sexual problems in women were libido 41%, arousal 29%, lubrication 56%, orgasm 35%, and dyspareunia 46%, and in men libido 47%, impotence 32%, partial impotence 52%, orgasm 41%, and ejaculation 43%. Both genders reported a negative body image. Patients seldom remembered discussing sexual risks preoperatively and seldom were treated for dysfunction. Current age (P < 0.001), surgical procedure (P = 0.003), and preoperative sexual activity (P = 0.001) were independently associated with current sexual activity. Gender (male, P = 0.014), surgical procedure (P = 0.005), and radiation therapy (P = 0.0001) were independently associated with the outcome "surgery made sexual life worse." Global QOL scores were high. CONCLUSIONS: Sexual problems after surgery for rectal cancer are common, multifactorial, inadequately discussed, and untreated. Therefore, sexual dysfunction should be discussed with rectal cancer patients, and efforts to prevent and treat it should be increased. PMID- 16041213 TI - Osteonectin influences growth and invasion of pancreatic cancer cells. AB - OBJECTIVE: We sought to examine the expression and functional role of osteonectin in primary and metastatic pancreatic ductal adenocarcinoma (PDAC). BACKGROUND: The glycoprotein osteonectin plays a vital role in cell-matrix interactions and is involved in various biologic processes. Overexpression of osteonectin is present in malignant tumors and correlates with disease progression and poor prognosis. METHODS: Expression of osteonectin was analyzed by quantitative polymerase chain reaction and immunohistochemistry in pancreatic tissues and by enzyme-linked immunosorbent assay in the serum of patients and donors. Recombinant osteonectin and specific antisense oligonucleotides were used to examine the effects of osteonectin on induction of target genes, and on proliferation and invasiveness of pancreatic cancer cells. RESULTS: There was a 31-fold increase in osteonectin mRNA levels in PDAC and a 16-fold increase in chronic pancreatitis as compared with the normal pancreas (P < 0.01). By immunohistochemistry, faint immunoreactivity was detected in the normal pancreas. In contrast, strong staining of the cancer cells was observed in addition to extensive osteonectin immunoreactivity in surrounding fibroblasts and in the extracellular matrix. In metastatic tissues, strong immunoreactivity was observed in fibroblasts and in extracellular matrix surrounding metastatic cancer cells, whereas the signal was absent in most tumor cells. In vitro studies showed that osteonectin was able to inhibit cancer cell growth while promoting invasiveness of pancreatic tumor cells. CONCLUSION: Osteonectin is markedly overexpressed in pancreatic cancer and has the potential to increase the invasiveness of pancreatic cancer cells. PMID- 16041214 TI - Positron emission tomography/computed tomography influences on the management of resectable pancreatic cancer and its cost-effectiveness. AB - OBJECTIVE: We sought to determine the impact of positron emission tomography/computed tomography (PET/CT) on the management of presumed resectable pancreatic cancer and to assess the cost of this new staging procedure. SUMMARY BACKGROUND DATA: PET using 18F-fluorodeoxyglucose (FDG) is increasingly used for the staging of pancreatic cancer, but anatomic information is limited. Integrated PET/CT enables optimal anatomic delineation of PET findings and identification of FDG-negative lesions on computed tomography (CT) images and might improve preoperative staging. MATERIAL AND METHODS: Patients with suspected pancreatic cancer who had a PET/CT between June 2001 to April 2004 were entered into a prospective database. Routine staging included abdominal CT, chest x-ray, and CA 19-9 measurement. FDG-PET/CT was conducted according to a standardized protocol, and findings were confirmed by histology. Cost benefit analysis was performed based on charged cost of PET/CT and pancreatic resection and included the time frame of staging and surgery. RESULTS: Fifty-nine patients with a median age of 61 years (range, 40-80 years) were included in this analysis. Fifty-one patients had lesions in the head and 8 in the tail of the pancreas. The positive and negative predictive values for pancreatic cancer were 91% and 64%, respectively. PET/CT detected additional distant metastases in 5 and synchronous rectal cancer in 2 patients. PET/CT findings changed the management in 16% of patients with pancreatic cancer deemed resectable after routine staging (P = 0.031) and was cost saving. CONCLUSIONS: PET/CT represents an important staging procedure prior to pancreatic resection for cancer, since it significantly improves patient selection and is cost-effective. PMID- 16041215 TI - Model for End-Stage Liver Disease (MELD) predicts nontransplant surgical mortality in patients with cirrhosis. AB - OBJECTIVE: We sought to determine the ability of the Model for End-Stage Liver Disease (MELD) score to predict 30-day postoperative mortality for patients with cirrhosis undergoing nontransplant surgical procedures. SUMMARY BACKGROUND DATA: The Child-Pugh class historically has been used by clinicians to assist in management decisions involving patients with cirrhosis. However, this classification scheme has a number of limitations. Recently, MELD was introduced. It has been shown to be highly predictive of mortality in a variety of clinical scenarios. METHODS: Adult patients with a diagnosis of cirrhosis undergoing nontransplant surgical procedures between January 1, 1996, and January 1, 2002, at a single center were analyzed. The preoperative MELD score was calculated for all patients, and the MELD's performance in predicting 30-day mortality was determined using multivariate regression techniques. RESULTS: A total of 140 surgical procedures were identified and analyzed. The 30-day mortality rate was 16.4%. The mean admission MELD score for the patients who died (23.3, 95% confidence interval 19.6-27.0) was significantly different from those patients surviving beyond 30 days (16.9, 15.6-18.2), P = 0.0003. The c-statistic for MELD score predicting 30-day mortality was 0.72. Further subgroup analysis of 67 intra abdominal surgeries showed an in-hospital mortality of 23.9%. The mean MELD score for patients dying (24.8, 20.4-29.3) was significantly different from survivors (16.2, 14.2-18.2), P = 0.0001. The c-statistic for this subgroup was 0.80. CONCLUSIONS: The MELD score, as an objective scale of disease severity in patients with cirrhosis, shows promise as being a useful preoperative predictor of surgical mortality risk. PMID- 16041216 TI - Prognostic impact of anatomic resection for hepatocellular carcinoma. AB - OBJECTIVES: To evaluate the prognostic impact of anatomic versus nonanatomic resection on the patients' survival after resection of a single hepatocellular carcinoma (HCC). SUMMARY OF BACKGROUND DATA: Anatomic resection is a reasonable treatment option for HCC; however, its clinical significance remains to be confirmed. METHODS: Curative hepatic resection was performed for a single HCC in 210 patients; the patients were classified into the anatomic resection (n = 156) and nonanatomic resection (n = 54) groups. In 84 patients assigned to the anatomic resection group, segmentectomy or subsegmentectomy was performed. We evaluated the outcome of anatomic resection, including segmentectomy and subsegmentectomy, in comparison with that of nonanatomic resection, by the multivariate analysis taking into consideration 14 other clinical factors. RESULTS: Both the 5-year overall survival and disease-free survival rates in the anatomic resection group were significantly better than those in the nonanatomic resection group (66% versus 35%, P = 0.01, and 34% versus 16%, P = 0.006, respectively). In the segmentectomy and subsegmentectomy group, the 5-year overall and disease-free survival rates were 67% and 28%, respectively, both of which were also higher than the corresponding rates in the nonanatomic resection group (P = 0.007 and P = 0.001, respectively). The results of multivariate analysis revealed that anatomic resection was a significantly favorable factor for overall and disease-free survivals: the hazard ratios were 0.57 (95% confidence interval, 0.32-0.99, P= 0.04), and 0.65 (0.43-0.96, P = 0.03). CONCLUSION: Anatomic resection for a single HCC yields more favorable results rather than nonanatomic resection. PMID- 16041217 TI - Role of postchemotherapy surgery in the management of patients with liver metastases from germ cell tumors. AB - OBJECTIVE: To evaluate the role of postchemotherapy adjunctive surgery in patients with liver metastases from germ cell cancer (GCT). PATIENTS AND METHODS: Forty-three male patients with nonseminoma were treated in different multicenter treatment protocols between 1990 and 1999, and they underwent hepatic surgery. The results of postchemotherapy surgical resection, histologic findings found during postchemotherapy surgery, and prognostic factors for survival were assessed. RESULTS: Thirty-five of 43 patients (81%) were initially diagnosed with liver metastases and advanced GCT, and 8 patients (19%) presented with metachronous liver metastases after a median interval of 16 months (range, 6-103 months). Twelve patients (28%) had isolated liver metastases after completion of chemotherapy, while 31 patients (72%) had additional residual extrahepatic tumor masses. Liver surgery included tumor excision or segmentectomy in 32 patients (74%) and hepatectomy (right/left) or resection of multiple segments in 11 patients (26%). Histologic analysis of postchemotherapy resected residua yielded necrosis in 67%, teratoma in 12%, and viable cancer in 21%. Additional resections at other sites have been performed in 31 patients revealing necrosis in 61% (n = 19), teratoma in 29% (n = 9), and vital carcinoma in 10% (n = 3). In 39% of patients, histologic findings differed among liver and other resection sites. Refractoriness to chemotherapy was associated with a shorter survival after surgery, and a trend was seen in patients with elevation of AFP. CONCLUSION: The high rate of viable cancer and teratoma found in liver specimens, differing histologic results at residual tumor locations, and the high survival rate achieved support a multidisciplinary approach including resection of liver masses since no accurate selection of patients can narrow the use of surgery. PMID- 16041218 TI - Left hepatic trisectionectomy for hepatobiliary malignancy: results and an appraisal of its current role. AB - OBJECTIVE: To analyze results of 70 patients undergoing left hepatic trisectionectomy and to clarify its current role. SUMMARY BACKGROUND DATA: Left hepatic trisectionectomy remains a complicated hepatectomy, and few reports have described the long-term results of the procedure. METHODS: Short-term and long term outcomes of 70 consecutive patients who underwent left hepatic trisectionectomy from January 1993 to February 2004 were analyzed. RESULTS: Of the 70 patients, 36 had colorectal liver metastasis, 24 had cholangiocarcinoma, 4 had hepatocellular carcinoma, and the remaining 6 had other tumors. Overall morbidity, 30-day and 90-day mortality rates were 46%, 7%, and 9%, respectively. Multivariate analysis disclosed that preoperative jaundice and intraoperative blood transfusion were positive independent predictors for postoperative morbidity; however, there were no independent predictors for postoperative mortality. Postoperative morbidity (87% versus 35%, P < 0.001) and mortality (20% versus 5%, P = 0.108) were observed more frequently in patients with preoperative obstructive jaundice than in those without jaundice. Each survival according to tumor type was acceptable compared with reported survivals. Survival for patients with colorectal liver metastasis undergoing left hepatic trisectionectomy with concomitant partial resection of the remnant liver was similar to those without this concomitant procedure. This concomitant procedure was not associated with postoperative morbidity and mortality. CONCLUSIONS: Left hepatic trisectionectomy remains a challenging procedure. Preoperative obstructive jaundice considerably increases perioperative risk. Concomitant partial resection of the remaining liver appears to be safe and offers the potential for cure in patients with colorectal metastasis affecting all liver segments. PMID- 16041219 TI - Patients' refusal of surgery strongly impairs breast cancer survival. AB - OBJECTIVE: To compare patient and tumor characteristics and survival between women who refused and women who accepted surgery for breast cancer. SUMMARY BACKGROUND DATA: Surgery represents the central component of curative breast cancer treatment, but some women decide not to undergo surgery. Recent studies on the prognosis of non operated breast cancer are nonexistent. PATIENTS AND METHODS: This study included all 5339 patients aged < 80 years with nonmetastatic breast cancer recorded at the Geneva Cancer Registry between 1975 and 2000. We consulted the clinical files of all nonoperated women to identify those who refused surgery. Patients who refused surgery were compared with those accepting surgery using logistic regression. The effect of refusal of surgery on breast cancer mortality was evaluated by Cox proportional hazards analysis. RESULTS: Seventy patients (1.3%) refused surgery. These women were older, more frequently single, and had larger tumors. Overall, 37 (53%) women had no treatment, 25 (36%) hormone-therapy alone, and 8 (11%) other adjuvant treatments alone or in combination. Five-year specific breast cancer survival of women who refused surgery was lower than that of those who accepted (72%, 95% confidence interval, 60%-84% versus 87%, 95% confidence interval, 86%-88%, respectively). After accounting for other prognostic factors including tumor characteristics and stage, women who refused surgery had a 2.1-fold (95% confidence interval, 1.5 3.1) increased risk to die of breast cancer compared with operated women. CONCLUSIONS: Women who refuse surgery for breast cancer have a strongly impaired survival. This information might help patients who are hesitant toward surgery make a better informed decision. PMID- 16041220 TI - Treatment trends in early-stage invasive lobular carcinoma: a report from the National Cancer Data Base. AB - OBJECTIVE: To examine treatment trends in invasive lobular carcinoma (ILC) over the last 15 years and, in particular, to compare rates of recurrence and disease free survival associated with breast conservation therapy compared with mastectomy. SUMMARY BACKGROUND DATA: The biologic characteristics of ILC make it difficult to estimate the extent of the disease by either clinical examination or mammography, and can also make it difficult to detect axillary lymph node metastases. Because of this, there has been a bias toward treating ILC with aggressive therapy. METHODS: Patients with ILC were selected from the National Cancer Data Base (1989-2001) using an extensive set of inclusion and exclusion criteria. A total of 21,596 patients were selected, including 8108 who received breast conservation therapy and 13,488 who received mastectomy. Analysis included demographic characteristics, trends in usage of sentinel lymph node biopsy, rates of local and distant recurrence, and 5-year disease-free survival rates. RESULTS: The use of breast conversation therapy increased almost threefold during the study period. From 1998 to 2001, the use of sentinel node biopsy increased more than twofold in the breast conservation group (an average of 23% in 1998 versus 57% in 2001), compared with limited usage in the mastectomy group (an average of 10% in 1998 versus 23% in 2001). Local recurrence rates were very low and disease free survival rates were correspondingly high in both treatment groups for all diagnosis years and across all pathologic tumor size/lymph node status designations. CONCLUSIONS: Less invasive treatment options are becoming widely used for invasive lobular carcinoma, yielding outcomes equivalent to those seen with more aggressive treatment. PMID- 16041221 TI - Surgery decreases long-term mortality, morbidity, and health care use in morbidly obese patients. PMID- 16041223 TI - A brief overview of bariatric surgical procedures currently being used to treat the obese patient. AB - Obesity has reached an overwhelming high in the United States as well as other developing countries around the world. Alone, approximately 60 million Americans are characterized as obese, and 10 million of them are considered morbidly obese. Many have tried and failed not only to lose the excess weight through fad diets, medically supervised diets, exercise programs, and athletic club memberships, but also to maintain a healthy weight. As a result, weight gain and loss has become a way of life, an unhealthy way of life. This often results in a weight gain of a greater number of pounds than where they started. As a result, many patients and their physicians are looking to weight loss surgery as a permanent solution to the problem of yo-yo dieting. Along with the surgical alterations to the digestive system, nutrition counseling with portion control and regular exercise are part of a comprehensive program for successful long-term weight maintenance. It is the intent of this article to provide the reader with the basic understanding of the normal anatomy of the digestive tract. Then, each surgical procedure will be discussed, enabling the reader to visualize the changes in the digestion and absorption of food. It is these changes in absorption of vitamins and minerals, fats and carbohydrates, and proteins that lead to the various medical complications seen in patients after bariatric surgery. PMID- 16041224 TI - Nutritional concerns related to Roux-en-Y gastric bypass: what every clinician needs to know. AB - Weight loss surgery, particularly the Roux-en-Y gastric bypass (REYGB), has become a popular treatment strategy for obesity. Often the only measure of success is the amount of weight lost following surgery. Unfortunately the nutritional adequacy of the postoperative diet has frequently been overlooked, and in the months to years that follow, nutritional deficiencies have become apparent, including protein-calorie malnutrition and various vitamin and mineral deficiencies contributing to medical illnesses and limiting optimal health. Therefore, patients require close monitoring following REYGB, with special regard to the rapidity of weight loss and vigilant screening for signs and symptoms of subclinical and clinical nutritional deficiencies. Several specific nutrients require close surveillance postoperatively to prevent life-threatening complications related to deficient states. This article addresses nutritional concerns associated with REYGB with fastidious focus on recognition and treatment of the nutritional deficiencies and promotion of nutritional health following REYGB. Recommendations regarding nutritional intake following REYGB are based on available scientific data, albeit limited. In cases where data do not exist, expert or consensus opinion is provided and recommendations for future research are given. Ultimately, clinical application of this information will contribute to the prevention of nutrition-related illness associated with REYGB. PMID- 16041226 TI - Hormonal status, fertility, and pregnancy before and after bariatric surgery. AB - At the present time, the exact etiology and developmental course of obesity in many individuals is still being debated. The majority of healthcare professionals are in agreement, however, that morbidly obese persons are at risk for a wide range of health problems, both before and after bariatric surgery. Obesity is, unfortunately, a condition in which the disease as well as the available treatments have the potential to cause serious alterations in health status. The average candidate for bariatric surgery has experienced decades of varying degrees of obesity as well as multiple failed dietary and medication regimens. As a result of almost continual physiologic instability, these individuals typically are subject to a wide range of hormonal imbalances. Since the majority of persons who are obese, and those seeking bariatric surgery, are women, it is important for healthcare providers to understand how these hormonal fluctuations can have a devastating impact on sexual and reproductive function. Although the research to date has been limited, studies have demonstrated changes in fertility, contraceptive response, and pregnancy outcomes in obese women, as well as obese women who undergo bariatric surgery. As morbid obesity continues to increase in incidence in the female population, greater numbers of women will seek bariatric surgery, and, as a consequence, experience some of the untoward effects shown to date. Addressing the concerns of these patients, both before and after surgery, will continue to be a complex and challenging task. PMID- 16041227 TI - The effects of weight loss on calcium and bone. AB - The article identifies 2 national healthcare problems that are occurring in the United States. The first national healthcare problem is obesity and the second is osteoporosis. What do these 2 healthcare problems have in common? They go hand in hand in teenagers and adults who have undergone weight reduction surgeries. These surgical procedures place a person at risk for osteoporosis because of the surgical procedure causing malabsorption that comes from the bypassing of the duodenum, which is the primary location for the absorption of calcium. This new high-risk population needs to become educated regarding osteoporosis and the treatment measures that can be instituted to prevent this disease. Osteoporosis is a disease where the bones of the body become very fragile and can easily break. Any bone can be affected by osteoporosis and fracture. There is no cure for osteoporosis but this disease can be controlled and prevented. This article discusses various treatment and prevention measures, ranging from dietary changes to the addition of medications. PMID- 16041228 TI - The application of laparoscopic bariatric surgery for treatment of severe obesity in adolescents using a multidisciplinary adolescent bariatric program. AB - The evolution of laparoscopic surgery has made bariatric surgery acceptable for weight loss; however, much controversy exists about its appropriateness for adolescents. Despite the controversial issues, the growing epidemic in adolescent obesity has resulted in rising numbers of applications for bariatric surgery. There are few bariatric surgical programs designed for adolescents. Pediatric settings face high start-up costs and poor reimbursement and lack established bariatric surgeons. Even so, bariatric surgery is increasingly being performed on adolescents in alarming numbers. To avoid adverse physical and psychosocial outcomes, the application of the principles of growth and development is essential. The program should be established as a multidisciplinary approach to management of adolescents and should be in institutions capable of meeting the guidelines for surgical treatment outlined by the American Society of Bariatric Surgery. To prevent postoperative complication, a multidisciplinary team of experienced medical and surgical specialists is needed for optimal preoperative decision making and postoperative management and long-term follow-up. Laparoscopic Roux-en-Y gastric bypass is a safe procedure and an effective means to treat obesity-related morbidity in the adolescent. Results have been excellent and justify a clinical trial to confirm the safety and efficacy of bariatric surgery in the adolescent population. PMID- 16041229 TI - My story: a personal perspective on bariatric surgery. AB - Because of the increased incidence of obesity today, bariatric surgery is now considered to be an acceptable treatment modality. However, much of the information that exists in the media focuses on the procedure itself, not on the preoperative and postoperative issues that can be just as important to address. As a certified registered nurse practitioner who deals with these patients on a daily basis, and as a patient who has had both the vertical banded gastroplasty and the Roux-en-Y gastric bypass, I consider myself as having a better understanding of the entire process than most. Everyone who chooses bariatric surgery has his or her own motivating factors, whether it be to feel better, to get off his or her medications, or to better control or resolve his or her comorbid conditions; the list is endless. The most important factor is educating oneself and knowing all there is to know about whatever procedure is desired. Insurance companies are now requiring nutritional consults preoperatively that lay the foundation for bariatric patients postoperatively. It is imperative as a healthcare consumer to ask questions, attend classes and support groups, and follow the guidelines that the surgeon has established so as to be successful. I have been through it all and understand the process from beginning to end. I also have extraordinary compassion, sensitivity, and empathy for the bariatric patients, because of my previous experiences. People who have never had a weight problem cannot begin to understand the plight of those who do. I try to guide the patients and point them in the right direction; however, the decisions that they make are ultimately theirs. PMID- 16041230 TI - Potential complications of obstructive sleep apnea in patients undergoing gastric bypass surgery. AB - Patients with morbid obesity undergoing gastric bypass surgery present many unique challenges to the healthcare team. Obstructive sleep apnea (OSA) is a common comorbidity in the postoperative gastric bypass patient. The complexity in the management of these patients during and after anesthesia is immense. Anesthetic agents and analgesics increase the severity of symptoms of OSA. Systemic opioid administration depresses the respiratory drive and decreases oxygen saturation. In addition, normal muscle hypotonia during rapid eye movement sleep increases the chance of respiratory complications during the initial postoperative period. Continuous positive airway pressure can effectively treat sleep apnea in the postoperative gastric bypass patient. There are concerns that pressurized air can inflate the stomach and intestines and result in the disruption of the anastomoses of the gastric bypass patient. If weight loss is sufficient, it can lead to an improvement or even cure of symptoms of OSA. PMID- 16041231 TI - Cultural competence. PMID- 16041232 TI - Application of cultural constructs in the care of first generation Latino clients in a community mental health setting. AB - As the Latino population in the United States grows, it is imperative to attend to the appropriateness of the mental health care that is being provided to its members. Latinos experience many of the same behavioral health disorders as other ethnic and cultural groups in the United States, but underutilize services relative to many other groups. Such underutilization may be related to issues such as stigma, language, and acculturation level, all of which often create barriers to treatment. First generation Latinos (i.e., individuals born outside the United States) are especially vulnerable to adverse experiences when seeking and receiving treatment. This may be due in part to acculturation and language issues which may further contribute to future underutilization of services. A well established therapeutic alliance developed through the appropriate use of cultural constructs may help mitigate some of the barriers faced by some Latino groups, especially those who are first generation. This paper reviews several cultural constructs that have been highlighted in the Hispanic behavioral health literature and discusses their potential implications for clinical care. This paper offers a number of practical clinical guidelines for mental health professionals who work with Latino groups. These clinical recommendations are based on a synthesis of selected cultural constructs and the clinical experiences of the authors' work in a large community-based Hispanic mental health clinic. PMID- 16041233 TI - Inter-rater and test-retest reliability of the Spanish language version of the eating disorder examination interview: clinical and research implications. AB - BACKGROUND: To examine the inter-rater and test-retest reliability of the Spanish Language version of the Eating Disorder Examination (S-EDE) in monolingual Latina women. Established measures are needed to study Latino groups, and short-term test-retest reliability findings are needed to provide context for clinical treatment and outcome studies. METHODS: Inter-rater reliability (IRR) and short term (5-14 days) test-retest reliability (TRR) of the S-EDE (using intraclass correlation coefficients [ICCs]) were examined in a non-clinical study group of 60 monolingual Latina women. RESULTS: IRR was excellent for objective bulimic episodes (ICC = 0.99) but was modest for subjective bulimic episodes (ICC = 0.55). TRR was good for objective bulimic episodes (ICC = 0.79) but was unacceptable for subjective bulimic episodes (ICC = 0.22). IRR and TRR kappa coefficients (0.56 and 0.37, respectively) for identifying the presence or absence of objective bulimic episodes were modest. For the S-EDE subscales, both IRR (ICCs ranged from 0.80 to 0.98) and TRR (ICCs ranged from 0.67 to 0.90) were good to excellent. CONCLUSIONS: These findings provide preliminary support for the reliability of the S-EDE for use with Latina women. The constructs of eating disorder psychopathology measured by the S-EDE subscales (restraint, eating concern, weight concern, and shape concern) and the core feature of binge eating (objective bulimic episodes) show high short-term consistency. The results for subjective bulimic episodes are consistent with previous studies that have questioned whether these eating behaviors are reliable indicators of eating disorders. Additional evaluation is needed with clinical groups. PMID- 16041234 TI - Aripiprazole: initial clinical experience with 142 hospitalized psychiatric patients. AB - BACKGROUND: Aripiprazole is the first dopamine D2 receptor partial-agonist approved for treatment of schizophrenia. Its apparently benign adverse-effect profile encourages broader use in other disorders, especially to limit weight gain associated with other antipsychotic or antimanic agents. We considered the first 6 months of experience with aripiprazole in psychiatric inpatients with a range of disorders. METHODS: We analyzed data obtained from medical records of patients treated with aripiprazole who were hospitalized at McLean Hospital (for 19 +/- 18 days) between December 2002 and June 2003 to evaluate dosing, tolerability, and clinical effects of this new agent in patients diagnosed with DSM-IV psychotic, major affective, or other disorders. RESULTS: Out of a sample of 2766 adult inpatients (65.5% women), 142 were given aripiprazole (mean final daily dose, 16.1 +/- 6.2 mg, 0.20 +/- 0.09 mg/kg body weight) for major affective disorders (52%), primary psychotic disorders (40%), and dementia (8%). CGI ratings improved by 20% on average. Adverse effects were infrequent (15.5%), were three times more likely among women, and most often involved moderate behavioral activation or nausea, with no new episodes of mania. Of the patients who were given aripiprazole, 83% continued it at discharge. Many patients were obese when they were admitted, and obesity was associated with relatively low mg/kg doses of aripiprazole. CONCLUSIONS: Aripiprazole was used in a range of disorders and was generally well tolerated. Adverse effects may reflect its unique dopamine partial agonist activity. Since aripiprazole is likely to be considered for obese patients, body weight should be considered in establishing adequate doses. Controlled trials of this antipsychotic agent in disorders other than schizophrenia are needed. PMID- 16041235 TI - Antipsychotic polypharmacy: is there evidence for its use? AB - Antipsychotic polypharmacy occurs frequently in clinical practice; however, there is a lack of controlled clinical studies testing the efficacy of the combinations used. The purpose of this literature review was to examine studies and other reports that have assessed the incremental benefits and deficits of combination antipsychotic therapy versus monotherapy. A PUBMED search covering a 26-year period from 1976 to 2002 was conducted. The search was limited to clinical trials, case series, and reports. Fifty-two reports were identified that systematically assessed the efficacy of combination therapy as opposed to monotherapy: 4 double-blind studies, 13 open-label clinical trials, and 35 case reports. Only one open-label trial and 2 case reports met the design criteria of having trials of each medication and the combination in the same patients and using some type of standardized assessment to evaluate outcome. The most frequent combination was clozapine-risperidone. Of the clinical trials, 75% (3/4) of the double-blind studies and 69% (9/13) of the open-label trials found that combination therapy was effective in reducing symptoms, while 37% (13/35) of case reports documented an overall positive outcome. Currently, the clinical practice of antipsychotic polypharmacy is not evidence-based; however, there is also no evidence against its use. Expanded systematic research to assess this clinical practice is needed. PMID- 16041236 TI - Clinical application of the concept of relative potency: an example involving chlorpromazine and haloperidol. PMID- 16041237 TI - Sex offender commitment in Illinois. PMID- 16041238 TI - What is the measure of a safe hospital? Medication errors missed by risk management, clinical staff, and surveyors. AB - Research in the last decade has identified medication errors as a more frequent cause of unintended harm than was previously thought. Inpatient medication errors and error-prone medication usage are detected internally by medication error reporting and externally through hospital licensing and accreditation surveys. A hospital's rate of medication errors is one of several measures of patient safety available to staff. However, prospective patients and other interested parties must rely upon licensing and accreditation scores, along with varying access to outcome data, as their sole measures of patient safety. We have previously reported that much higher rates of medication errors were found when an independent audit was used compared with rates determined by the usual process of self-report. In this study, we summarize these earlier findings and then compare the error detection sensitivity of licensing and accreditation surveys with that of an independent audit. When experienced surveyors fail to detect a highly error prone medication usage system, it raises questions about the validity of survey scores as a measure of safety (i.e., lack of medication errors). Replication of our findings in other hospital settings is needed. We also recommend measures for improving patient safety by reducing error rates and increasing error detection. PMID- 16041239 TI - Influence of ABCB1, ABCC1, ABCC2, and ABCG2 haplotypes on the cellular exposure of nelfinavir in vivo. AB - OBJECTIVES: The human immunodeficiency virus protease inhibitor nelfinavir is substrate of polyspecific drug transporters encoded by ABCB1 (P-glycoprotein), ABCC1 (MRP1) and ABCC2 (MRP2), and an inhibitor of BCRP, encoded by ABCG2. Genetic polymorphism in these genes may be associated with changes in transport function. METHODS: A comprehensive evaluation of single nucleotide polymorphisms (39 SNPs in ABCB1, 7 in ABCC1, 27 in ABCC2, and 16 in ABCG2), and inferred haplotypes was done to assess possible associations of genetic variants with cellular exposure of nelfinavir in vivo. Analysis used peripheral mononuclear cells from individuals receiving nelfinavir (n=28). Key results were re-examined in a larger sample size (n=129) contributing data on plasma drug levels. RESULTS AND CONCLUSIONS: There was no significant association between cellular nelfinavir area under the curve (AUC) and SNPs or haplotypes at ABCC1, ABCC2, ABCG2. There was an association with cellular exposure for two loci in strong linkage disequilibrium: ABCB1 3435C>T; AUCTT>AUCCT>AUCCC (ratio 2.1, 1.4, 1, Ptrend=0.01), and intron 26 +80T>C; AUCCC> AUCCT > AUCTT (ratio 2.4, 1.3, 1, Ptrend=0.006). Haplotypic analysis using tagging SNPs did not improve the single SNP association values. PMID- 16041240 TI - Three haplotypes associated with CYP2A6 phenotypes in Caucasians. AB - The human cytochrome P450 2A6 (CYP2A6) enzyme metabolizes several xenobiotic compounds of clinical or toxicological importance. We aimed to identify genetic variants and major CYP2A6 haplotypes associated with CYP2A6 phenotypic variation. CYP2A6 mRNA level, protein level, activity and haplotypes were determined in Caucasian liver samples via real-time polymerase chain reaction, Western blot, coumarin 7-hydroxylation, DNA sequencing and genotyping, respectively. Phenotypes were then analyzed for associations with haplotypes. CYP2A6 transcript, protein and activity levels were correlated among each other. In 45 African-American, 156 Caucasian, 47 Chinese, 50 Japanese and 47 Korean DNA samples, we detected 95 different polymorphisms in the CYP2A6 gene, 49 of which had not been described previously. Caucasian variants formed 33 haplotypes which built four clades. Allele *9B and the CYP2A7/2A6 partial deletion allele CYP2A6*12B were both associated with decreased expression. The latter haplotype extends at least over 147 kb up into the CYP2B6 gene. A haplotype almost identical to allele *1A was associated with decreased expression and activity of CYP2A6 compared to all other haplotypes. In summary A CYP2A6*1A-like allele, *9B and *12B are major genetic determinants of CYP2A6 phenotype variation in Caucasians. PMID- 16041241 TI - CYP3A7 protein expression is high in a fraction of adult human livers and partially associated with the CYP3A7*1C allele. AB - Previously, cytochrome P450 3A7 (CYP3A7), which constitutes the major CYP enzyme in fetal livers, has been considered a fetus-specific enzyme. However, CYP3A7 mRNA has recently been shown to be expressed at significant levels in a subset of adult human livers, several of which carry the CYP3A7*1C allele that contains the proximal PXR/CAR element of CYP3A4. The objective of this study was to investigate CYP3A7 expression at the protein level by developing a CYP3A7 specific antibody to allow its quantification. Based on results from 59 adult human liver samples, we found significant CYP3A7 protein expression in approximately one in 10 adult livers amounting for 24-90 pmol/mg microsomal protein, thereby contributing 9-36% to total CYP3A levels in these livers. CYP3A7 protein was detected in five of seven livers carrying the CYP3A7*1C allele (two of which only had trace amounts), whereas an additional three livers expressing CYP3A7 were apparently homozygous for CYP3A7*1. The mean protein expression level of CYP3A7 was 42 pmol/mg within the group of livers expressing CYP3A7 and 4 pmol/mg in all liver samples. CYP3A7 expression was thus higher than that of the polymorphically expressed CYP3A5 in adult human livers, based on a comparison with a previous study using our CYP3A5 peptide-specific antibody. The relatively high level of CYP3A7 protein expression detected in a subset of adult livers may be relevant with respect to the metabolism of exogenous and endogenous substrates, such as retinoic acid and dehydroepiandrosterone. PMID- 16041242 TI - 164Ile allele in the beta2-Adrenergic receptor gene is associated with risk of elevated blood pressure in women. The Copenhagen City Heart Study. AB - OBJECTIVE: Since beta2-adrenergic receptors are important regulators of blood pressure, genetic variation in this receptor could explain risk of elevated blood pressure in selected individuals. We tested the hypothesis that Gly16Arg, Gln27Glu, and Thr164Ile in the beta2-adrenergic receptor gene associated with elevated blood pressure. METHODS: We genotyped 9185 individuals from the adult Danish general population. RESULTS: Allele frequencies of 16Arg, 27Glu, and 164Ile were 0.38, 0.44, and 0.01, respectively. Among women never treated with antihypertensive medication those heterozygous for Thr164Ile versus non-carriers had increased diastolic blood pressure (P=0.02). Women heterozygous for Thr164Ile versus non-carriers had an odds ratio for elevated blood pressure of 1.93 (95% CI: 1.30-2.86). Finally, women double heterozygous for Thr164Ile and Gln27Glu or Gly16Arg versus non-carriers at all 3 loci had an odds ratio for elevated blood pressure of 2.49 (1.28-4.85) or 3.19 (1.46-6.97). In men, blood pressure was not influenced by this genetic variation. CONCLUSION: In women Thr164Ile heterozygosity is associated with increased diastolic blood pressure, and represent a risk factor for elevated blood pressure in women in the general population. This was most pronounced in those women also heterozygous for Gln27Glu or Gly16Arg. PMID- 16041243 TI - Functional characterization of non-synonymous single nucleotide polymorphisms in the gene encoding human multidrug resistance protein 1 (MRP1/ABCC1). AB - The 190-kDa ATP-binding cassette (ABC) multidrug resistance protein 1 (MRP1) encoded by the MRP1/ABCC1 gene mediates the active cellular efflux of glucuronide, glutathione and sulfate conjugates. It can also confer resistance to a diverse spectrum of chemotherapeutic agents and transport a variety of toxicants. In the present study, we examined 10 MRP1/ABCC1 missense genetic variants [non-synonymous single nucleotide polymorphisms (SNPs)] to determine whether or not they affect expression or function of the transporter. Variants 218C>T (Thr73Ile), 257C>T (Ser92Phe), 350C>T (Thr117Met), 689G>A (Arg230Gln), 1898G>A (Arg633Gln), 2168G>A (Arg723Gln), 2965G>A (Ala989Thr), 3140G>C (Cys1047Ser), 3173G>A (Arg1058Gln) and 4535C>T (Ser1512Leu) were recreated using site-directed mutagenesis and transfected into human embryonic kidney cells. Immunoblotting experiments showed that all mutant proteins were expressed at levels comparable to wild-type MRP1. Vesicular transport assays revealed that the Ala989Thr mutation caused a significant decrease in estradiol 17beta-glucuronide transport due to a decrease in apparent affinity (Km) for this organic anion. The transport properties of the other mutants were comparable to wild-type MRP1. When the MRP1/ABCC1 non-synonymous SNPs were evaluated by the SIFT algorithm using subsets of homologs and orthologs of MRP1/ABCC1, Arg230Gln, Val353Met, Arg433Ser, Gly671Val and Arg1058 mutations were predicted to be deleterious, whereas the PolyPhen algorithm predicted Ser92Phe and Gly671Val to be potentially damaging. Thus most predictions of these algorithms were not in accordance with our experimental results. In conclusion, our data suggest that none of the MRP1/ABCC1 variants studied are likely by themselves to have major deleterious effects in healthy individuals, and the SIFT and PolyPhen algorithms appear to be poor predictors of the phenotypic consequences of these MRP1 mutations at least in vitro. PMID- 16041244 TI - Dopamine transporter (SLC6A3) 5' region haplotypes significantly affect transcriptional activity in vitro but are not associated with Parkinson's disease. AB - The dopamine transporter (DAT) plays a critical role in dopaminergic neurotransmission and is also the major site of action for some drugs of abuse. The coding region of the DAT gene, SLC6A3, is well conserved, but non-coding regions are more variable, most notably a variable number of tandem repeats (VNTR) polymorphism in the 3' untranslated region, which has been studied in a number of dopamine-related neurological disorders, including Parkinson's disease (PD). We aimed to characterize variation in the 5' region of SLC6A3 because little is known about the extent of variation in this region and potential consequences of such variation on gene expression. We identified multiple single nucleotide polymorphisms (SNPs) covering approximately 5000 bp 5' of exon 1 through the start of exon 2 (+2106). These SNPs segregated as eight haplotypes, six of which were common. These haplotypes differed significantly in activity in a reporter gene activity assay. However, we did not observe associations between common SNPs or haplotypes and PD in a case-control study of 261 incident cases and 376 age- and gender-matched unrelated controls. By contrast, we did observe a modest association of the 3' VNTR 9-repeat allele with PD (odds ratio=1.45; 95% confidence interval=1.04-2.03). This association was limited to subjects 60 years of age and greater versus those less than 60 years of age. We conclude that although DAT 5' region SNPs haplotypes significantly alter in vitro transcriptional activity, they are not related to PD risk. In addition, our findings provide further evidence supporting an association of PD with the VNTR polymorphism. PMID- 16041245 TI - Influence of the eNOS gene on development of blood pressure and left ventricular mass: longitudinal findings in multiethnic youth. AB - OBJECTIVES: To evaluate the impact of the endothelial nitric oxide synthase (eNOS) gene on longitudinal development of blood pressure (BP) and left ventricular mass (LVM) from childhood into early adulthood. METHODS: Three polymorphisms including -922A>G, intron 4VNTR, and Glu298Asp of the eNOS gene were investigated. Individual growth-curve modeling and haplotype trend regression analyses were conducted for 579 white and black American youths with 12 assessments over a 15-year period. RESULTS: Significantly different allele and genotype frequencies were observed between blacks and whites for all three polymorphisms. Linkage disequilibrium (LD) patterns among these polymorphisms were also different between ethnic groups: strong LD between the -922A>G and intron 4 VNTR loci was observed in whites but not in blacks. Single locus analyses identified a significant interaction between the intron 4 VNTR and gender on diastolic BP (DBP) levels. The 4a allele carriers had significantly lower DBP levels in males (P=0.012), but higher DBP levels in females (P=0.045). Haplotype analyses confirmed the DBP lowering effect in males (P=0.049). DBP in males homozygous for haplotype G-4a-Glu was 2.58 mmHg lower than males homozygous for the most common haplotype (A-non4a-Glu). Additionally, individuals homozygous for haplotype G-non4a-Glu showed a 0.51 mmHg steeper increase in DBP per year with age as compared to the most common haplotype (P=0.007). No associations between single polymorphisms or haplotypes of the eNOS gene and systolic BP or LVM were found. CONCLUSIONS: our results suggest that eNOS gene may have gender specific and age-dependent effects on DBP and the development of hypertension risk. PMID- 16041246 TI - Cost-effectiveness of hepatitis A/B vaccination in the private sector. PMID- 16041247 TI - High prevalence of sexually transmitted infections among female sex workers in the eastern highlands province of Papua New Guinea: correlates and recommendations. AB - BACKGROUND: More than 200 female sex workers (FSWs) participating in commercial sex along the Highlands Highway of Papua New Guinea were identified in a previous survey. This has implications for the spread of sexually transmitted infections (STIs) and human immunodeficiency virus (HIV) to areas and population groups serviced by the road. GOAL: The goal of this study was to estimate the prevalence of gonorrhea, chlamydia, syphilis, trichomoniasis, and HIV among FSWs in Goroka and Kainantu in the Eastern Highlands Province (EHP) and to identify correlates that could be considered in intervention and control. STUDY: Self-identified FSWs recruited through the Goroka Sex Workers Peer-Mediated Programme were invited to participate. All consenting FSWs underwent pretest counseling and provided sociodemographic and behavioral data using a structured questionnaire. The women were also asked to self-collect vaginal specimens and to provide peripheral blood to detect the respective STIs and HIV. RESULTS: Results were available for 211 FSWs. None of the women were positive for HIV. The overall estimated rates for gonorrhea, chlamydia, syphilis, and trichomoniasis were 21%, 19%, 24%, and 51%, respectively. Seventy-four percent were positive for at least 1 STI and 43% had multiple STI infections. High-risk sexual behaviors were found to be common among the women, including low and inconsistent use of condoms, with most of them attributing this to unavailability, dislike by or familiarity with clients, and being drunk and/or high on marijuana. CONCLUSIONS: STIs are prevalent among FSWs in Goroka and Kainantu in the EHP and are maintained by widespread high-risk sexual behaviors, including low use of condoms. Implications for their spread through the highway warrants increased efforts in intervention. Apart from a need to promote condom acceptance, distribution, and use, other high-risk sexual behavior and correlates identified in this study provide important considerations for intervention and control in this population. PMID- 16041248 TI - Costs and effects of chlamydial screening: dynamic versus static modeling. AB - OBJECTIVE: The objective of this study was to assess the impact of modeling type on the economic evaluation of screening programs for asymptomatic Chlamydia trachomatis infections. STUDY: We compared a stochastic network simulation model (dynamic model) with a decision analysis model (static model) for estimating the cost-effectiveness of an opportunistic screening program. The influence of the model type on the required data, the computed results, and the sensitivity of model parameters were investigated. RESULTS: When compared with static modeling, dynamic modeling yielded different cost-effectiveness ratios and identified other optimal screening strategies as it considers changes in the force of infection caused by screening. However, it is more complex, data- and time-demanding, and more sensitive to some parameters affecting the force of infection than static modeling. CONCLUSIONS: Dynamic models should be applied for the economic evaluation of prevention measures that have the potential to lower the force of infection such as large-scale chlamydial screening programs. PMID- 16041249 TI - Enhanced erythrocyte aggregation in clinically diagnosed pelvic inflammatory disease. AB - OBJECTIVE: Enhanced erythrocyte aggregation, revealed using a simple bedside test, has been found recently in several inflammatory conditions. The diagnosis of pelvic inflammatory disease (PID) is at times difficult because of the vague symptoms and signs, but is crucial because even mild PID can have future reproductive consequences. Our objective was to determine the degree of erythrocyte aggregation in PID. The demonstration of an increase in aggregation could be of additive value in cases in which the diagnosis is difficult. STUDY DESIGN: A prospective case-control study was conducted. Fifteen consecutive women diagnosed clinically as having PID based on Centers for Disease Control and Prevention criteria, and 15 matched controls were enrolled. Blood samples were drawn for hematologic indices, C-reactive protein, fibrinogen levels, and red cell aggregation. We studied the degree of red cell aggregation using a simple slide test and image analysis. The variable measured was the erythrocyte percent (EP), equivalent to the slide area covered by erythrocytes. RESULTS: Erythrocyte percent was 59.6 +/- 4.2 and 80.0 +/- 3.6 for the study group and controls, respectively (P <0.001). A significant difference was noted also for the other hematologic and biochemical markers of inflammation between patients and the controls. CONCLUSIONS: We conclude that the degree of erythrocyte aggregation is increased in PID. Its simplicity, rapidity, and low cost should be further evaluated as a diagnostic tool in the context of this frequent disease. PMID- 16041250 TI - HIV-related risk behavior among Hispanic immigrant men in a population-based household survey in low-income neighborhoods of northern California. AB - OBJECTIVES/GOAL: We compared risk behaviors and HIV testing between recent (in the U.S. <5 years) and established (in the U.S. >5 years) Hispanic immigrant men (N = 410). STUDY: This study was a population-based, cross-sectional survey of HIV/sexually transmitted disease markers and risk behaviors in men age 18 to 35 years residing in low-income census block groups in 3 northern California counties. RESULTS: Recent immigrants were less likely to currently have a main sexual partner (45.3% vs. 67.2%, P <0.01) and more likely to have ever used commercial sex workers (40.0% vs. 27.6%, P <0.01). Recent immigrants were less likely to receive medical care in the last 6 months (21.2% vs. 31.3%, P = 0.04) or had ever been HIV tested (26.0% vs. 43.3%, P <0.01). Established immigrants more likely reported unprotected sex, hallucinogen or ecstasy use. CONCLUSIONS: Recent Hispanic immigrants have less stable sexual partnerships and less health seeking behavior, including HIV testing. Established immigrants report HIV test rates comparable to the national average. PMID- 16041251 TI - Why nonoxynol-9 may have failed to prevent acquisition of Neisseria gonorrhoeae in clinical trials. AB - OBJECTIVES: To understand why clinical trials failed to demonstrate efficacy of nonoxynol-9 in preventing gonorrhea. GOAL: To test the hypothesis that nonoyxnol 9 failed to prevent acquisition of Neisseria gonorrhoeae because most isolates are resistant to killing by nonoyxnol-9 at the level attainable with intravaginal use. STUDY: The lowest concentrations of nonoxynol-9 required to kill or inhibit growth of clinical isolates of N gonorrhoeae and Lactobacillus were determined. RESULTS: Most strains (17 of 25) of N gonorrhoeae (68%) were resistant to the highest concentration of nonoxynol-9 tested (20%). L crispatus (100%), L jensenii (90%), and L iners (79%) were also resistant to nonoxynol-9. CONCLUSIONS: N gonorrhoeae and H2O2-producing strains of vaginal lactobacilli were not killed by nonoxynol-9 at concentrations greater than those achievable in vivo. Earlier studies that formed the basis for subsequent trials most likely did not detect resistance because too few isolates were evaluated. Large numbers of clinical isolates should be examined before the initiation of clinical trial using microbicidal products. PMID- 16041252 TI - A study evaluating ceftriaxone as a treatment agent for primary and secondary syphilis in pregnancy. AB - OBJECTIVES: To evaluate the efficacy of ceftriaxone in pregnant women who were diagnosed with early syphilis. STUDY: Eleven women with a history of penicillin allergy, positive skin test, but prior history of safe usage of cephalosporins were included. Ceftriaxone (250 mg) was given intramuscularly once daily for 7 and 10 days to patients with primary and secondary syphilis, respectively. A second course of therapy was provided at 28 weeks' gestation. The rapid plasma reagin test (RPR) was measured before and after therapy. The blood of neonates was also tested at delivery and during the follow-up period. RESULTS: The serum RPR titers of 11 mothers decreased fourfold, 3 months after treatment. Ten patients developed negative RPR results. The serum RPR was negative at delivery or 6 months after delivery in all neonates. CONCLUSIONS: Ceftriaxone may be considered as an alternative for treatment of early syphilis in pregnancy. PMID- 16041253 TI - Epidemiology of genital herpes simplex virus infections in a community-based sample in France: results of the HERPIMAX study. AB - OBJECTIVE: The objective of this study was to provide information on the prevalence of herpes simplex infections in the general population in Europe. GOALS: The goals of this study were to determine the prevalence of clinically probable genital herpes and the relationship between serotype and clinical expression in a French community-based sample. STUDY: A total of 4410 subjects chosen at random were serotyped for herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2). Data on symptoms were obtained by questionnaire allowing retrospective diagnosis of clinically probable genital herpes. RESULTS: Questionnaire data and serotype were available for 3192 subjects. Seroprevalences of HSV-1 and HSV-2 were 65.6% and 15.5%, respectively. Prevalence of clinically probable genital herpes was 11.8%, identified in 11.1% of HSV-1-positive subjects and 26.8% of HSV-2-positive subjects, with a lower prevalence in those coinfected with both virus types. CONCLUSIONS: Clinically probable genital herpes was observed in one fourth of subjects with HSV-2 infections and in some subjects with HSV-1 infection. Coinfection with HSV-1 appeared to protect against symptom expression in subjects infected with HSV-2. PMID- 16041254 TI - Social and geographic distance in HIV risk. AB - OBJECTIVE: The objective of this study was to examine the relationship between social distance (measured as the geodesic, or shortest distance, between 2 people in a connected network) and geographic distance (measured as the actual distance between them in kilometers [km]). STUDY: We used data from a study of 595 persons at risk for HIV and their sexual and drug-using partners (total N = 8920 unique individuals) conducted in Colorado Springs, Colorado, from 1988 to 1992--a longitudinal cohort study that ascertained sociodemographic, clinical, behavioral, and network information about participants. We used place of residence as the geographic marker and calculated distance between people grouped by various characteristics of interest. RESULTS: Fifty-two percent of all dyads were separated by a distance of 4 km or less. The closest pairs were persons who both shared needles and had sexual contact (mean = 3.2 km), and HIV-positive persons and their contacts (mean = 2.9). The most distant pairs were prostitutes and their paying partners (mean = 6.1 km). In a connected subset of 348 respondents, almost half the persons were between 3 and 6 steps from each other in the social network and were separated by a distance of 2 to 8 km. Using block group centroids, the mean distance between all persons in Colorado Springs was 12.4 km compared with a mean distance of 5.4 km between all dyads in this study (P <0.0001). The subgroup of HIV-positive people and their contacts was drawn in real space on a map of Colorado Springs and revealed tight clustering of this group in the downtown area. CONCLUSION: The association of social and geographic distance in an urban group of people at risk for HIV provides demonstration of the importance of geographic clustering in the potential transmission of HIV. The proximity of persons connected within a network, but not necessarily known to each other, suggests that a high probability of partner selection from within the group may be an important factor in maintenance of HIV endemicity. PMID- 16041255 TI - Accounting for failures may improve precision: evidence supporting improved validity of self-reported condom use. AB - OBJECTIVES: To determine whether a measure of unprotected vaginal sex that is adjusted for condom failures would produce improved accuracy in predicting biologically confirmed STDs (chlamydia and gonorrhea) among female teens. METHODS: Self-reported measures were collected using audio-computer-assisted self interviewing. DNA amplification for the presence of Chlamydia trachomatis and Neisseria gonorrhoeae was conducted. RESULTS: The unadjusted measure of unprotected vaginal sex was not significantly associated with biologically confirmed prevalence of STDs (prevalence ratio [PR] = 1.51; 95% CI = 0.71-3.21; P = 0.28). Alternatively, the adjusted measure achieved significance (PR = 3.59; 95% CI = 1.13-11.38; P = 0.014). More than one quarter (25.6%) of teens using condoms inconsistently and/or incorrectly tested positive for an STD compared to 7.1% among those reporting the consistent and correct use of condoms. CONCLUSION: Findings demonstrate that studies of condom effectiveness should use an adjusted measure of condom use to achieve precision and rigor. PMID- 16041256 TI - HIV/AIDS risk among brothel-based female sex workers in China: assessing the terms, content, and knowledge of sex work. PMID- 16041257 TI - Herpes simplex virus type 2 status at age 26 is not related to early circumcision in a birth cohort. AB - OBJECTIVE: The objective of this study was to determine if circumcision in early childhood affects the risk of acquiring herpes simplex virus type 2 (HSV-2) infection. STUDY: Study members were born in 1972-1973 in Dunedin, New Zealand. Circumcision status was sought at age 3, when the cohort was established. Information about sexual behavior was obtained at ages 21 and 26. Serum was tested for HSV-2 antibodies at age 26 for 435 men (82.9% of the surviving cohort). RESULTS: Of eligible men, 40.2% had been circumcised. The prevalence of HSV-2 antibodies was 7.3% in uncircumcised men and 7.4% in circumcised men. Social and sexual factors were very similar between the 2 groups and adjustment had no effect on the association (odds ratio, 1.1; 95% confidence interval, 0.46 2.5). Seroconversion rates according to years since first sexual intercourse were 0.85 and 0.86 per 100 person-years for uncircumcised and circumcised men. CONCLUSION: The results support a lack of association between circumcision status and HSV-2 acquisition, although a small effect cannot be ruled out. PMID- 16041258 TI - Management of cytomegalovirus infection in lung transplant recipients: evidence based recommendations. AB - Cytomegalovirus (CMV) continues to cause significant morbidity and mortality in lung transplant recipients. This article presents recommendations based on available evidence for the optimal management of CMV in lung transplant recipients, which have been developed by an expert committee of transplant physicians-surgeons and infectious disease specialists. PMID- 16041259 TI - Steroids can be safely withdrawn from cyclosporine and mycophenolate mofetil treated renal allograft recipients: long-term results. AB - BACKGROUND: Discontinuation of steroids has long been a goal of transplant teams. However, whether this strategy is associated or not with a higher risk of long term graft loss has not been resolved. METHODS: The authors analyzed a cohort of 91 renal allograft recipients who underwent transplantation between 1993 and 1997. They were treated with cyclosporine and mycophenolate mofetil (MMF) and then had steroids withdrawn. Inclusion criteria were as follows: serum creatinine lower than 133 microM, first or second renal transplants, no or only one acute rejection episode (borderline or Ia grade), and a peak of panel reactive antibodies under 50%. Prednisone was gradually tapered off and then discontinued over a period of 2 to 4 months. RESULTS: There were no episodes of acute rejection after steroid withdrawal. Whether steroids were withdrawn before (early) or after (late) 6 months of renal transplantation did not influence outcome. By Kaplan-Meier analysis, patient survival was 93.6% and 100% at 5 years and 93.6% and 97.6% at 10 years in the early and late steroid withdrawal groups, respectively. Graft survival was 94.3% and 98.1% at 5 years and 87.6% and 82.4% at 10 years in the early and late steroid-withdrawal groups, respectively. Risk factors for graft loss in multivariate analysis were peak of panel reactive antibodies (relative risk, 1.074; 95% confidence interval, 1.017-1.134; P=0.01) and acute rejection (relative risk, 16.5; 95% confidence interval, 1.8-147; P=0.01). CONCLUSIONS: Early and late steroid withdrawal in low-immunologic-risk renal allografts treated with cyclosporine and MMF can be achieved without risk of acute rejection and with excellent long-term results. PMID- 16041260 TI - Thrombotic microangiopathy in living-donor liver transplantation. AB - BACKGROUND: Thrombotic microangiopathy (TMA) is a fatal complication characterized by microvascular occlusive disorder resulting in systemic or intrarenal platelet aggregation, severe thrombocytopenia, and microangiopathic hemolytic anemia. Only sporadic case reports of TMA after cadaveric or living donor liver transplantation (LDLT) have been described. METHODS: The authors report 10 (5% of the total series) TMA patients after LDLT and review the previously reported cases. TMA was diagnosed on the basis of progressive thrombocytopenia of unknown cause and microangiopathic hemolytic anemia, suggested by sharp elevation of serum lactate dehydrogenase levels and the presence of fractionated erythrocytes in blood smear. RESULTS: Of the 10 patients with TMA, 7 presented with viral hepatitis (2 with hepatitis B and 5 with hepatitis C virus infection) as the cause of end-stage liver disease. Clinical diagnosis of TMA was made at a median interval of 18 days (range, 3-356 days) from the time of transplantation. Conversion of calcineurin inhibitors (CNI) was conducted in nine patients. One patient recovered after CNI conversion alone. Plasma exchange was performed in eight patients. Three patients died. CONCLUSIONS: Immediate treatment of TMA should be initiated by reduction or conversion of CNI followed by plasma exchange. Hepatitis C virus infection might contribute to the high incidence of TMA in LDLT patients. PMID- 16041261 TI - Characteristic pattern of chromosomal imbalances in posttransplantation lymphoproliferative disorders: correlation with histopathological subcategories and EBV status. AB - BACKGROUND: Posttransplantation lymphoproliferative disorders (PTLDs) are a spectrum of lymphoid proliferations, occurring in immunosuppressed organ transplant recipients. They comprise early lesions, polymorphic (P-PTLD), monomorphic (M-PTLD), and Hodgkin/Hodgkin-like lymphoma PTLD (HL-PTLD) lesions. Most of them are associated with Epstein-Barr virus (EBV). Little is known about their genetic changes. MATERIALS AND METHODS: We have studied 35 PTLDs[7 P-PTLDs (3/7 polyclonal IgH), 26 M-PTLDs (22 B-cell PTLD, 4 T-cell PTLD), 2 HL-PTLDs], using comparative genomic hybridization (CGH), a DNA-based technique allowing a screening of chromosomal imbalances without needing cultured cells. RESULTS.: Overall incidence of chromosomal imbalances: 51.5 %. The most frequent gains involved 8q24, 3q27 [4 cases each]; 2p24p25, 5p, 9q22q34, 11, 12q22q24, 14q32, 17q, 18q21 [2 cases each]. Nonrandom losses were 17p13 [4 cases]; 1p36, 4q [3 cases each]; 17q23q25, Xp [2 cases each]. Three high-level amplifications were detected: 4p16, 9p22p24, 18q21q23. In this latter imbalance, involvement of Bcl2 has been confirmed by FISH. The nonrandom CGH imbalances occurring in M-PTLD are usually described in lymphomas of immunocompetent patients and contain genes known to be involved in lymphomagenesis, while genomic abnormalities detected in half cases of EBV positive P-PTLD are mostly unknown. CONCLUSION: This study reported nonrandom chromosomal imbalances in PTLD and also identified early genomic alterations in EBV positive P-PTLD. These results raise two questions: the role of such lesions in the development and progression of those EBV induced lymphoproliferations and their clinical significance especially in P-PTLD. PMID- 16041262 TI - Longitudinal comparisons of lymphocytes and subtypes between airway wall and bronchoalveolar lavage after human lung transplantation. AB - BACKGROUND: T lymphocytes are crucial in lung allorejection. The contribution of lymphocyte subtypes to the pathogenesis of chronic rejection (bronchiolitis obliterans syndrome [BOS]) remains unclear. METHODS: Twenty-nine initially healthy lung transplant recipients underwent 136 bronchoscopic assessments, including bronchoalveolar lavage (BAL) (with flow cytometry) and endobronchial biopsies (EBB) (with immunohistochemistry) over 3 years of follow-up. RESULTS: Of the 29 patients studied over 3 years, 23 developed BOS category 0 p and 17 went on to BOS 1. Compared with controls, the BAL percentage of CD4 cells was lower and the percentage of CD8 cells was increased significantly early posttransplant. Subsequent BAL lymphocyte subtype changes with time, or with the development of BOS, were minimal. By contrast, the early posttransplant EBB lymphocyte numbers were normal (P>0.05 vs. controls); subsequently, CD3 and CD8 (but not CD4) cells were increased with time in patients who did not develop BOS (P<0.05) and, more strikingly, in patients who eventually developed BOS (P<0.01). Multivariate analyses suggested an association between BAL lymphocytes (percentage) and azathioprine dose, female gender, rejection grade A on transbronchial biopsies, and pre-BOS status, whereas EBB CD8 cell counts were associated with time posttransplant, pretransplant diagnosis, and rejection grade B on TBB. CONCLUSIONS: There is an early, persistent low percentage of BAL CD4 T cells, high BAL CD8 T cells, and progressively increasing airway wall CD3 and CD8 T cells with time posttransplant in healthy patients (but more predominantly in BOS patients) after transplantation. These immunopathologic changes may suggest that CD8 T cells could escape current immunosuppression and participate in chronic lung rejection. PMID- 16041263 TI - Lymphoproliferative disease after renal transplantation in Australia and New Zealand. AB - BACKGROUND: Lymphoproliferative disease is a common and serious complication of organ transplantation. It is well documented that the risk of its development increases with the level of immunosuppression. Less is known about its incidence, prevalence, timing, and prognosis. METHODS: The authors conducted a retrospective review of all patients with lymphoproliferative disease after renal transplantation documented in the Australia and New Zealand Dialysis and Transplant Registry from 1970 to March 2003. RESULTS: One hundred ninety-seven cases of lymphoproliferative disease occurred in 15,930 allografts in 13,516 recipients. There has been a steady increase in its incidence and prevalence each decade since 1970. Cases cluster into an early group (<2 years after transplantation) and a late group (5-10 years after transplantation). Risk factors include exposure to a calcineurin inhibitor, but there was no increased risk in those treated with anti-T-lymphocyte antibodies. Patient survival was poor: 51% at 1 year and 39% at 5 years. CONCLUSIONS: Lymphoproliferative disease is an increasingly common problem after renal transplantation, and the outcome is poor. Measures to reduce its incidence might include reduction of long-term immunosuppression exposure. Established disease has a high short-term mortality, and new treatment options, such as anti-B-lymphocyte monoclonal antibodies, should be aggressively pursued. PMID- 16041264 TI - Co-infection of polyomavirus-BK and cytomegalovirus in renal transplant recipients. AB - BACKGROUND: Polyomavirus-BK (BK) is a significant cause of allograft dysfunction in renal transplant recipients. Cytomegalovirus (CMV) and BK infection are thought to be possible risk factors for one another, but no supporting data are yet available. METHODS: The authors monitored BK and CMV infection by quantitative polymerase chain reaction (PCR) in 69 renal transplant recipients with serum creatinine elevation to determine the prevalence of co-infection. In addition, 150 adult renal transplant recipients were also retrospectively analyzed for both infections. RESULTS: Of 69 recipients, 12 were plasma BK-PCR positive. Eight of the 12 showed high BK levels (>10 copies) and BK nephropathy. Six of the 12 were also CMV-PCR-positive compared with only 3 of 57 plasma BK negative patients (50% vs. 5.3%, P=0.001). Comparatively, the incidence of Epstein-Barr virus infection was similar in both groups (1 of 12 [8.3%] vs. 2 of 57 [3.5%], P =not significant). In addition, retrospective analysis of CMV-PCR positivity in 150 adult renal transplant recipients showed similar results (5 of 6 in BK-PCR-positive [83%] vs. 8 of 144 in BK-PCR-negative [5.6%], P=0.00001). More plasma BK-PCR-positive patients had concomitant CMV infection than CMV-PCR positive patients with BK infection (5 of 6 [83%] vs. 4 of 13 [31%], P=0.05). CONCLUSIONS: In conclusion, high plasma BK-positivity (>10) is significantly associated with BK nephropathy. Plasma BK-positivity is highly associated with co infection of CMV, suggesting possible risk factors for one another. Therefore, detection of either infection strongly suggests the need to monitor for the other. This strategy may lead to the prevention of virus-induced complications by preemptive antiviral therapy in renal allografts. PMID- 16041265 TI - Bone marrow-derived stem-cell repopulation contributes minimally to the Type II pneumocyte pool in transplanted human lungs. AB - BACKGROUND: Lung transplant recipients are vulnerable to immunologic, infectious, ischemic, and toxic pulmonary injuries. The authors investigated whether type II pneumocytes in the lungs of cross-gender lung transplant patients show genotypic evidence to support repopulation of the lung by stem cells of bone marrow origin, and whether the degree of repopulation was related to rejection history. METHODS: Recut sections were obtained from lung biopsy specimens from seven male recipients of transplanted lungs from female donors. Sequential immunohistochemistry and fluorescence in situ hybridization was performed on each section to evaluate for Y-chromosome-containing type II pneumocytes. RESULTS: Y chromosome-containing type II pneumocytes were found in 9 of 25 biopsy specimens from 5 of 7 gender-mismatched male lung transplant recipients, and accounted for 0% to 0.553% of type II pneumocytes. There was no evidence of polyploidy to suggest cell-cell fusion. The number of type II pneumocytes of male karyotype showed a statistically significant relationship to the cumulative number of episodes of acute cellular rejection. CONCLUSIONS: Lung transplant recipients develop low levels of pneumocyte repopulation by bone marrow-derived stem cells or their progeny. These cells contribute minimally to the type II pneumocyte proliferation that is often present in these patients as a sequela to alveolar injury. PMID- 16041266 TI - A radical scavenger, edaravone, protects canine kidneys from ischemia-reperfusion injury after 72 hours of cold preservation and autotransplantation. AB - BACKGROUND: Cold ischemia-reperfusion (I/R) injury is a prominent cause of delayed graft function after kidney transplantation. Reactive oxygen species play a crucial role in I/R injury. Edaravone is a synthetic radical scavenger that has been used in acute stroke. Some animal experiments have revealed its beneficial effects against I/R injury, but its effects after cold preservation and transplantation of canine kidneys are unknown. METHODS: Female hybrid dogs weighing 11 to 13 kg were used. Under anesthesia, the left kidney was harvested. After 72 hr of preservation in cold histidine-tryptophan-ketoglutarate solution, autotransplantation was performed in the right iliac fossa, with contralateral nephrectomy. Animals were divided into control and treatment groups (n=6 per group). In the treatment group, edaravone was administered intravenously at harvest and at reperfusion (3 mg/kg) and in addition was added to the preservation solution (50 microM). RESULTS: Animal survival at 2 weeks was four of six in the control group and six of six in the treatment group. Compared with controls, treated animals had higher mean urine output, higher mean glomerular filtration rate, improved tubular cell function, lower mean serum creatinine, and lower renal vascular resistance. Biopsy specimens from treated animals showed less tubular cell damage and decreased P-selectin expression in endothelial cells. Lipid peroxidation of renal tissue and urinary excretion of 8-hydroxy-2' deoxyguanosine were suppressed by the treatment. CONCLUSIONS: Edaravone reduced cold I/R injury in canine renal transplantation. The agent has the potential to ameliorate preservation injury in clinical transplantation. PMID- 16041267 TI - Expression of biologically active human TRAIL in transgenic pigs. AB - BACKGROUND: Xenotransplantation of porcine organs into human recipients is a potential option for overcoming the dramatic shortage of suitable donor organs. To date, transgenic modification of pig organs has achieved partial or temporal reduction of xenograft rejection by inhibition of hyperacute rejection. Expression of human tumor necrosis factor-alpha-related apoptosis-inducing ligand (TRAIL) in transgenic pigs might be a strategy for controlling posthyperacute rejection mechanisms mediated by cellular components of the immune system. The objective of this study was generation of a transgenic pig model to evaluate the potential of this strategy for xenotransplantation. METHODS: Transgenic pigs were generated by microinjection of an expression vector for human TRAIL under control of the murine H-2K promoter. Expression of the transgene was analyzed by Western blot and immunohistochemistry. Biologic activity of TRAIL on transgenic porcine lymphocytes was evaluated in co-culture experiments using Jurkat and Hut 78.2 cells as targets. RESULTS: In three lines of transgenic pigs, human TRAIL protein was detected in the membrane fractions of various tissues. Highest expression levels were observed in spleen and lung. Human TRAIL expression on porcine lymphocytes was augmented on activation of cells. Transgenic pig lymphoblasts induced apoptosis in Jurkat and Hut 78.2 cells, which was inhibited by neutralizing anti-TRAIL antibodies, demonstrating a TRAIL-specific effect. CONCLUSIONS: Ubiquitous expression of human TRAIL was achieved in transgenic pigs without detrimental side effects. Pigs expressing biologically active human TRAIL will be used for future xenotransplantation experiments to modulate primate anti pig cellular immune responses. PMID- 16041268 TI - Effect of a novel immunosuppressant, ST1959, on the immune system and renal allograft survival in rats. AB - BACKGROUND: ST1959 is a 3,5-diaryl-s-triazole belonging to a novel class of contragestional agents with immunosuppressant activity. The aim of the present study was to investigate the effects of this drug on allogeneic immune response and on renal allograft survival in rats. METHODS: One group of naive and one group of allosensitized Lewis rats received ST1959 (0.5 mg/kg/day for 6 days administered subcutaneously). The respective control groups received vehicle alone. At the end of treatment, all rats were killed and thymus, spleen, lymph nodes, bone marrow, and blood were harvested. Cell number, leukocyte subpopulations, and lymphocyte alloreactivity were evaluated. Three additional groups of Lewis rats received an allogeneic (Brown Norway [BN]) kidney transplant: two groups received ST1959 (0.5 mg/kg daily until death or for 6 days and then twice weekly), and the last one received vehicle. RESULTS: In naive rats, ST1959 reduced the percentage of CD4CD8 (74.2+/-2.7%; vehicle, 89.1+/-1.1%; P<0.05) and increased the percentage of CD4CD8 thymocytes (5.7+/-0.8% vs. 2.8+/ 0.4%; P<0.05). Infusion of allogeneic (BN) splenocytes caused a twofold increase of activated CD4 T cells (CD4CD25) that was prevented by ST1959 treatment. Consistently, the alloreactivity of lymphocytes from naive and allosensitized animals treated with ST1959 was significantly lower than that of control rats. ST1959 (in both tested regimens) significantly prolonged renal allograft survival in comparison with vehicle (12.4+/-0.5 vs. 7.7+/-0.5 days; P<0.001). CONCLUSIONS: ST1959 possesses immunomodulatory effects and significantly prolongs survival of renal allografts in rats. PMID- 16041269 TI - Short-term immunosuppression facilitates induction of mixed chimerism and tolerance after bone marrow transplantation without cytoreductive conditioning. AB - BACKGROUND: Induction of mixed chimerism and tolerance usually requires cytoreduction or transplantation of high numbers of bone marrow cells (BMC). However, such protocols have only a suboptimal success rate and, more importantly, equivalent numbers of BMC cannot be routinely obtained in the clinical setting. The authors therefore evaluated whether a short-course of immunosuppression (IS) given in addition to co-stimulation blockade would facilitate chimerism induction and allow reduction of the minimally required number of BMC without cytoreduction. METHODS: B6 mice received 200, 100, or 50 x 10 unseparated BMC from Balb/c donors plus an anti-CD40L monoclonal antibody (mAb) and CTLA4Ig (without irradiation or cytotoxic drugs). Some groups were treated additionally with IS (rapamycin, methylprednisolone, and mycophenolate mofetil for 4 weeks after bone marrow transplantation), donor-specific transfusion (DST), or anti-OX40L mAb, as indicated. RESULTS: IS led to long-term multilineage chimerism in 9 of 10 mice receiving 200 x 10 BMC (without IS, 1 of 4; P<0.05), in all mice (n=10) receiving 100 x 10 (without IS, 6 of 9; P<0.05), and notably in 9 of 10 mice treated with 50 x 10 BMC (without IS, 4 of 10; P<0.05). With transient IS, donor skin grafts were accepted longer than 170 days in 9 of 10 mice receiving 200 x 10 (without IS, 0 of 5 mice; P<0.05), all mice receiving 100 x 10 (without IS, 6 of 9; P<0.05), and 6 of 11 mice receiving 50 x 10 BMC (without IS, 4 of 10). The use of DST or anti-OX40L mAb had no beneficial effect. CONCLUSIONS: Transient IS significantly improves rates of chimerism and donor skin graft survival, and allows lasting mixed chimerism after transplantation of only 50 x 10 BMC. Thus, IS might help in the further development of noncytoreductive chimerism protocols. PMID- 16041270 TI - Everolimus versus mycophenolate mofetil in the prevention of rejection in de novo renal transplant recipients: a 3-year randomized, multicenter, phase III study. AB - BACKGROUND: This 36-month, randomized, parallel-group study compared safety and efficacy of two doses of everolimus with mycophenolate mofetil (MMF) in de novo renal-transplant recipients. METHODS: Renal-allograft recipients received 1.5 mg/day or 3 mg/day of everolimus or 2 g/day of MMF, plus full-dose cyclosporine (CsA) and corticosteroids after randomization. For at least their first year, patients received study medication according to a double-blinded, double-dummy design. Concerns over nephrotoxicity led to a protocol amendment to an open-label design with reduced CsA troughs. RESULTS: Incidences of primary efficacy failure at 36 months (biopsy-proven acute rejection, graft loss, death, or loss to follow up) were everolimus 1.5 mg/day, 33.7% (65/193); everolimus 3 mg/day, 34.0% (66/194); and MMF, 31.1% (61/196) (P=0.810). Antibody-treated acute rejection at 36 months was significantly lower with everolimus 1.5 mg (9.8%) than MMF (18.4%, P=0.014). Discontinuation for adverse events was more frequent with everolimus and hemolytic uremic syndrome, lymphoproliferative disease, and proteinuria, and higher serum creatinine occurred at increased frequency relative to the MMF arm. Creatinine levels in the everolimus arms were stable in follow-up: the mean rise in creatinine over the first 6 months of the open-label phase was 3 micromol/L or greater with everolimus and 7 micromol/L with MMF. However, serum creatinine levels were lower in the MMF group throughout. Death and graft loss were higher in the everolimus arms (not significant). CONCLUSIONS: As part of triple-drug immunosuppression, everolimus (1.5 or 3 mg/day) was as efficacious as MMF, although the side-effect profile featured increased adverse events. Nephrotoxicity/calcineurin-inhibitor-related adverse events will require judicious lowering of CsA exposure with monitoring of everolimus troughs. PMID- 16041271 TI - Inhibition of chronic rejection and development of tolerogenic T cells after ICOS ICOSL and CD40-CD40L co-stimulation blockade. PMID- 16041272 TI - Inhibition of chronic rejection and development of tolerogenic T cells after ICOS ICOSL and CD40-CD40L co-stimulation blockade. AB - BACKGROUND: : Blockade of the CD40-CD40L pathway results in long-term allograft survival but does not prevent chronic rejection. ICOS-ICOSL are members of the CD28-B7 family that play an important role in T-cell activation. METHODS: : The authors analyzed the effect of single or combined treatment with an anti-ICOS monoclonal antibody and CD40 immunoglobulin (Ig) on acute and chronic rejection of heart allografts in rats. RESULTS: : Treatment with anti-ICOS resulted in a modest but significant prolongation of allograft survival. Treatment with CD40Ig resulted in long-term graft survival but the cardiac grafts developed chronic rejection lesions. Combined CD40Ig+anti-ICOS treatment led to indefinite graft survival in all recipients and a significant decrease of chronic rejection lesions compared with CD40Ig alone. Importantly, four of the seven CD40Ig+anti ICOS-treated recipients showed a complete absence of chronic rejection lesions, whereas all of the CD40Ig-treated recipients showed chronic rejection. The CD40Ig+anti-ICOS group also showed significant decreased graft infiltration, decreased antidonor cytotoxic T-lymphocyte activity, and decreased alloantibodies compared with the CD40Ig-treated group. Adoptive transfer of splenocytes indefinitely prolonged allograft survival, whereas those depleted of T cells did not, suggesting the development of T-regulatory mechanisms. CONCLUSIONS: : These data indicate that the chronic rejection mechanisms that are CD40-CD40L independent are ICOS-ICOSL dependent. These results were obtained with conservation of cognate immune responses and development of tolerogenic T cells. PMID- 16041273 TI - Anti-intercellular adhesion molecule-1 antibodies in sera of heart transplant recipients: a role in endothelial cell activation. AB - BACKGROUND: Antiendothelial antibodies to non-human leukocyte antigens are made by a subset of heart transplant recipients, but the specificity of such antibodies is undefined. Intercellular adhesion molecule (ICAM)-1 is an abundantly expressed adhesion molecule with polymorphic residues, expressed on the surface of endothelial cells. The hypothesis that ICAM-1 acts as a minor histocompatibility antigen and that anti-ICAM-1 antibodies, directed against polymorphic residues, could be one component of the antiendothelial antibodies found after heart transplantation has been tested. METHODS: Chinese hamster ovary cells were transfected with full-length polymorphic variants of human ICAM-1. The binding of antibodies (immunoglobulin [Ig] G or IgM) to these cells was measured using sera from 50 heart transplant recipients (pretransplant and 1 and 2 years posttransplant) and sera from 20 normal volunteers by flow cytometry. The recipients and donors were genotyped for ICAM-1 polymorphisms. RESULTS: Sixty eight percent (n=34) of patients made IgM antibodies that bound to ICAM-1. However, it seems unlikely that ICAM-1 is a minor transplantation antigen, because there were no differences in antibody production from recipients matched or mismatched for ICAM-1 alleles. The antibodies bound to mouse endothelial cells that were engineered to overexpress human ICAM-1, and induced a robust activation of the Erk-2 mitogen-activated protein kinase pathway. CONCLUSIONS: Anti-ICAM-1 antibodies are produced after cardiac transplantation, but not to polymorphic residues. Such antibodies may contribute to the endothelial activation by binding to the endothelium, causing activation of proinflammatory signaling pathways. PMID- 16041274 TI - Half-life analysis of pancreas and kidney transplants. AB - Although graft and patient survival data are available for pancreas and kidney transplants, they are rarely reported in terms of half-life. Our aim was to determine whether a more relevant measure of outcome is patient and allograft half-life. Using the data from the Organ Procurement and Transplantation Network Registry on kidney and pancreas transplants from January 1988 to December 1996, patient and graft half-life and 95% confidence intervals were calculated and demographic variables compared. No significant differences were found between demographic variables. Kidneys transplanted in diabetics as a simultaneous kidney pancreas (SPK) fared better than diabetics receiving a kidney alone (9.6 vs. 6.3 years). Pancreatic graft survival in an SPK pair was better than pancreas after kidney transplant or pancreas transplant alone (11.2 vs. 2.5 years). Because kidney and pancreatic grafts have a longer half-life when transplanted with their mate grafts, we should consider the relative benefits of SPKs over pancreas after kidney transplant or pancreas transplant alone to limit the loss of precious resources. PMID- 16041275 TI - Negative impact of human leukocyte antigen matching in the outcome of polyomavirus nephropathy. AB - Factors dependent on the host, the virus, and the allograft affect the course of polyomavirus allograft nephropathy (PVAN). Development of specific cytotoxic antiviral immunity requires recognition of host human leukocyte antigen (HLA) molecules together with viral peptides on the target cells. We correlated the number of matched HLA-A, HLA-B, and HLA-DR antigens with graft outcome in 90 patients with PVAN. Patients that maintained graft function had significantly less degrees of HLA matching (mean 1.5) in comparison to patients with graft loss (mean 2.87, P= 0.001). Markedly reduced incidence of graft loss was observed in patients with no HLA matching whatsoever in comparison to patients with any degree of matching (P= 0.003). Lack of HLA matching may impair the host's ability to mount an effective antiviral cytotoxic immune response. An adequately developed antiviral cellular immunity may lead to significant tissue damage and graft loss even if the viral infection is eventually controlled. PMID- 16041276 TI - Polyclonal antibody-induced serum sickness in renal transplant recipients: treatment with therapeutic plasma exchange. AB - Serum sickness is an immune-complex mediated illness that frequently occurs in patients after polyclonal antibody therapy (ATGAM or thymoglobulin). Serum sickness presents with significant morbidity but is self-limited and resolves with prolonged steroid therapy. We present five renal transplant patients who developed serum sickness after polyclonal antibody treatment with severe symptoms that persisted after being started on systemic steroids. These patients underwent one or two courses of therapeutic plasma exchange (TPE) with subsequent complete resolution of their symptoms. Renal transplant recipients with serum sickness after polyclonal antibody therapy may benefit from TPE by accelerating their time to recovery and thereby reducing overall morbidity. PMID- 16041277 TI - In utero transplantation of human cord blood cells into rabbits. PMID- 16041278 TI - Successful liver transplantation in two cases of metastatic gastrointestinal stromal tumors. PMID- 16041279 TI - Tissue remodeling in a bioartificial fibromuscular patch following transplantation in a human. PMID- 16041280 TI - Page kidney causing reversible acute renal failure: an unusual complication of transplant biopsy. PMID- 16041281 TI - Living donor liver transplantation from donor with previous upper abdominal surgery. PMID- 16041282 TI - Schistosoma mansoni infection and liver graft. PMID- 16041283 TI - Venous thromboembolism in renal transplant recipients. PMID- 16041285 TI - Qualitative MRI: evidence of usual aging in the brain. AB - The understanding of aging in our society with steadily increasing life expectancy is an important challenge in medical science since socioeconomic pressure increases in parallel. Magnetic resonance imaging is a most useful tool to explore age related changes in the central nervous system especially in the brain. This article will focus on current knowledge and importance of such changes. Special attention will paid to white matter hyperintensities in terms of occurrence and progression and cerebral microbleeds in terms of their association to various diseases and their possible influence on thrombolytic therapy. Furthermore the meaning of darkening of the basal ganglia will be discussed and, in more general terms, the occurrence of virchow robin spaces and changes in cerebral metabolites assessable by proton magnetic resonance spectroscopy. PMID- 16041286 TI - Image processing: global and regional changes with age. AB - Our knowledge about the process of aging has continued to evolve as the methods used to study this process become more sophisticated. As more becomes known about the diagnostic criteria for dementia, the population of subjects taking part in aging studies has become more carefully screened minimizing the role of dementia as a confounding variable. Furthermore, advances in imaging techniques now allow us to view the anatomy of the brain in vivo better than any time in the past paving the way for longitudinal studies of the brain. It should not be surprising given the changes seen in studies of aging that our conventional wisdom of the aging process is being called into question. PMID- 16041287 TI - Quantitative MRI: hidden age-related changes in brain tissue. AB - The advent of MRI has made a remarkable progress in the understanding of age related brain changes providing a noninvasive tool to study in vivo the normally aging individuals at multiple time points. However, conventional MRI techniques are unable to detect and quantify age-related microstructural changes that have been documented at the post-mortem examination of brain tissues. More sophisticated, quantitative MR techniques such as magnetization transfer imaging, diffusion tensor imaging, and proton MR spectroscopy have been shown to be sensitive to microstructural and metabolic changes that occur in gray and white matter over the course of life span. This review highlights some of these innovative, quantitative MR techniques that are particularly relevant for the study of occult age-related brain tissue changes. Characterization of the in vivo patterns of molecular and cellular changes that occur in the normal aging brain is of crucial importance to understand the pathophysiology of normal cognitive decline and to interpret observed changes in neurodegenerative diseases. PMID- 16041288 TI - Epidemiology of white matter lesions. AB - This is an overview of the prevalence of, and risk factors for white matter lesions in older persons. The prevalence of white matter lesions is high and increases with age. Most risk factors that have been examined in association with white matter lesions are similar to those for cardiovascular disease, including elevated blood pressure, diabetes atherosclerosis, homocysteine levels, and markers of oxidative stress. The genetic contributions to white matter lesions are also reviewed. PMID- 16041289 TI - Pathological aging of the brain: an overview. AB - The number of elderly people is increasing rapidly and, therefore, an increase in neurodegenerative and cerebrovascular disorders causing dementia is expected. Alzheimer disease (AD) is the most common cause of dementia. Vascular dementia, dementia with Lewy bodies, and frontotemporal dementia are the most frequent causes after AD, but a large proportion of patients have a combination of degenerative and vascular brain pathology. Characteristic magnetic resonance (MR) imaging findings can contribute to the identification of different diseases causing dementia. The MR imaging protocol should include axial T2-weighted images (T2-WI), axial fluid-attenuated inversion recovery (FLAIR) or proton density weighted images, and axial gradient-echo T2*-weighted images, for the detection of cerebrovascular pathology. Structural neuroimaging in dementia is focused on detection of brain atrophy, especially in the medial temporal lobe, for which coronal high resolution T1-weighted images perpendicular to the long axis of the temporal lobe are extremely important. Single photon emission computed tomography and positron emission tomography may have added value in the diagnosis of dementia and may become more important in the future, due to the development of radioligands for in vivo detection of AD pathology. New functional MR techniques and serial volumetric imaging studies to identify subtle brain abnormalities may also provide surrogate markers for pathologic processes that occur in diseases causing dementia and, in conjunction with clinical evaluation, may enable a more rigorous and early diagnosis, approaching the accuracy of neuropathology. PMID- 16041290 TI - Sad seasons: winter mood not always holiday blues. PMID- 16041291 TI - Suffer the children: the psychobiology of early adversity. PMID- 16041292 TI - The impact of comorbidity on the management of pathological gambling. AB - A 30-year-old woman with severe pathological gambling and cyclothymia presented to our program with no previous history of pharmacologic or psychotherapeutic treatment. Pathological gambling is an impulse -control disorder not otherwise specified (Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition) in which comorbidity is common, particularly with substance abuse, obsessive compulsive disorder and mood disorders. As described in this case, pathological gamblers with bipolar comorbidity may be effectively treated with mood stabilizers such as lithium. After receiving 10 weeks of lithium treatment, the patient showed improvement in both gambling behavior and affective instability. The identification of specific subtypes among patients with pathological gambling may be relevant to the choice of pharmacologic treatment. PMID- 16041293 TI - Is SAD Lost to SAD? PMID- 16041294 TI - The diagnosis, symptomatology, and epidemiology of seasonal affective disorder. AB - The operational criteria for seasonal affective disorder (SAD) have undergone several changes since first proposed in 1984. SAD is currently included as a specifier of either bipolar or recurrent major depressive disorder in the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition. The International Classification of Diseases, Tenth Edition has provisional diagnostic criteria for SAD. The most characteristic quality of SAD is that the symptoms usually present during winter and remit in the spring. Furthermore, the symptoms tend to remit when the patients are exposed to daylight or bright light therapy. The cognitive and emotional symptoms are as in other types of depression but the vegetative symptoms are the reverse of classic depressive vegetative symptoms, namely increased sleep and increased appetite. SAD is a common condition, but the exact prevalence rates vary between different studies and countries and is consistently found to be more common in women and in youth. SAD probably possibly occurs in children although not as commonly as in young adults. Some studies have found that certain ethnic groups who live at high northern latitudes may have adapted to the long arctic winter. PMID- 16041295 TI - Update on the biology of seasonal affective disorder. AB - The etiology and pathophysiology of seasonal affective disorder (SAD) has been linked to the seasons and to light since its first conceptualization. Aspects of SAD that make it particularly amenable to biological investigation include the predictable recurrent episodes, the rapid response to a nonpharmacologic treatment, the specific neurovegetative features, and the availability of rich animal models of seasonality. This paper reviews new findings for the major biological hypotheses for SAD, focusing on circadian rhythms, neurotransmitters, and molecular genetics. Integrative issues and future directions for the study of SAD, including the heuristic value of a dual-vulnerability hypothesis that conceptualizes seasonality as a dimensional construct and the importance of studying endophenotypes, will be discussed. PMID- 16041296 TI - Light therapy for seasonal and nonseasonal depression: efficacy, protocol, safety, and side effects. AB - Bright light therapy for seasonal affective disorder (SAD) has been investigated and applied for over 20 years. Physicians and clinicians are increasingly confident that bright light therapy is a potent, specifically active, nonpharmaceutical treatment modality. Indeed, the domain of light treatment is moving beyond SAD, to nonseasonal depression (unipolar and bipolar), seasonal flare-ups of bulimia nervosa, circadian sleep phase disorders, and more. Light therapy is simple to deliver to outpatients and inpatients alike, although the optimum dosing of light and treatment time of day requires individual adjustment. The side-effect profile is favorable in comparison with medications, although the clinician must remain vigilant about emergent hypomania and autonomic hyperactivation, especially during the first few days of treatment. Importantly, light therapy provides a compatible adjunct to antidepressant medication, which can result in accelerated improvement and fewer residual symptoms. PMID- 16041297 TI - Pharmacotherapy of seasonal affective disorder. AB - Seasonal affective disorder is a common variant of recurrent major depressive disorder or bipolar disorder. Treatment with bright artificial light has been found to be effective in this condition. However, for patients who do not respond to light therapy or those who lack compliance, conventional drug treatment with antidepressants also has been proposed. Substances with selective serotonergic or noradrenergic mechanisms should be preferred over older antidepressants. Although there are a number of open and controlled studies evaluating different compounds, these studies were often limited by relatively small sample sizes. Furthermore, there are no studies specifically addressing bipolar seasonal depression. This article will review the published literature on pharmacotherapy of seasonal affective disorder. PMID- 16041306 TI - Blood and marrow transplant clinical trials network toxicity committee consensus summary: thrombotic microangiopathy after hematopoietic stem cell transplantation. AB - The syndrome of microangiopathic hemolysis associated with renal failure, neurologic impairment, or both is a recognized complication of hematopoietic stem cell transplantation. This entity is often called hemolytic uremic syndrome (HUS) or thrombotic thrombocytopenic purpura (TTP), yet it is clear that the pathophysiology of transplant-associated HUS/TTP is different from that of classic HUS or TTP. Furthermore, the incidence of this syndrome varies from 0.5% to 76% in different transplant series, primarily because of the lack of a uniform definition. The toxicity committee of the Blood and Marrow Transplant Clinical Trials Network has reviewed the current literature on transplant-related HUS/TTP and recommends that it be henceforth renamed posttransplantation thrombotic microangiopathy (TMA). An operational definition for TMA based on the presence of microangiopathic hemolysis and renal and/or neurologic dysfunction is proposed. The primary intervention after diagnosis of TMA should be withdrawal of calcineurin inhibitors. Plasma exchange, although frequently used in this condition, has not been proven to be effective. In the absence of definitive trials, plasma exchange cannot be considered a standard of care for TMA. It is hoped that these positions will improve the identification and reporting of this devastating complication after hematopoietic stem cell transplantation and facilitate future clinical studies for its prevention and treatment. PMID- 16041307 TI - Effector cells derived from host CD8 memory T cells mediate rapid resistance against minor histocompatibility antigen-mismatched allogeneic marrow grafts without participation of perforin, Fas ligand, and the simultaneous inhibition of 3 tumor necrosis factor family effector pathways. AB - Reduced-intensity conditioning regimens for transplant recipients have heightened awareness of immunologic resistance to allogeneic bone marrow transplants (BMT). Although T cell-mediated cytotoxicity has been assumed to play a role in the resistance against donor allogeneic hematopoietic stem and progenitor cell grafts, several studies have reported relatively unimpaired resistance by recipients who lack perforin, Fas ligand (FasL), and other cytotoxic mediators. This study compared the early kinetics of T cell-mediated resistance in B6 (H2b) cytotoxically normal versus deficient recipients after transplantation with major histocompatibility complex-matched, minor histocompatibility antigen (MiHA) mismatched allogeneic marrow grafts. Wild-type B6 or cytotoxic double-deficient perforin-/-/gld+/+ (B6-cdd) mice were sensitized against major histocompatibility complex-matched BALB.B or C3H.SW (H2b) MiHA and transplanted with a high dose (1 x 10(7)) of T cell-depleted bone marrow. CD8 T memory cells were shown to be present in recipients before BMT, and anti-CD8 monoclonal antibody infusion abolished resistance, thus demonstrating that CD8 T cells are the host effector population. Donor-committed and high proliferative potential progenitor numbers were markedly diminished by 48 hours after transplantation in both wild-type B6 and B6-cdd anti-donor MiHA-sensitized recipients. These observations indicate that the resistance pathway used in the cytotoxic deficient mice was both potent and rapidly induced--consistent with a CD8 memory T-cell response. To examine the role of Tumor necrosis factor-like weak inducer of apoptosis (TWEAK)- and TL1A mediated cytotoxicity in this strong resistance, newly generated monoclonal antibodies specific for these ligands were administered to B6-cdd recipients sensitized to donor antigens. Recipients of syngeneic B6-gfp bone marrow exhibited significant donor colony-forming unit numbers after BMT. In contrast, low or absent colony-forming unit levels were detected in allogeneic recipients, including those that lacked perforin and FasL and that received anti-TWEAK, anti tumor necrosis factor-related apoptosis-inducing ligand, and anti-TL1A monoclonal antibodies. These findings extend previous observations by demonstrating the existence of a rapidly effected resistance pathway mediated by memory CD8 effector T cells independent of the 2 major pathways of cytotoxicity. Together with previous findings, these results support the notion that effector cells derived from memory CD8 T-cell populations can mediate strong resistance against donor allogeneic MiHA-disparate hematopoietic engraftment by using a mechanism that is independent of the contribution of perforin, FasL, and the known death ligand receptor pathways. PMID- 16041308 TI - Long-term use of oral beclomethasone dipropionate for the treatment of gastrointestinal graft-versus-host disease. AB - Treatment of severe acute and chronic gastrointestinal (GI) graft-versus-host disease (GVHD) with prolonged high-dose systemic corticosteroids has limited success and considerable toxicity. Beclomethasone dipropionate (BDP) is a potent topically active steroid. We treated 15 patients with acute (n = 2) or chronic (n = 13) GI GVHD refractory to systemic corticosteroids with 28-day courses of oral BDP (2 mg 4 times daily). Response was measured by the change in GI score (sum of 6 GI symptoms) as well as the ability to taper or discontinue systemic corticosteroids. Nine (60%) of 15 evaluable patients responded to BDP, including 3 complete responses (a GI score of 0 or 1 and discontinuation of systemic corticosteroids). Attempts to taper calcineurin inhibitor during BDP therapy were unsuccessful. The 2 patients with acute GVHD had no response to BDP. Responders received a median of 3 cycles (range, 1-20), compared with 1 cycle (range, 1-5) in nonresponders. Suppression of the hypothalamic-adrenal axis was seen in 2 of the 5 patients tested, but neither demonstrated clinically significant symptoms. We conclude that BDP is safe and effective for long-term treatment of chronic GI GVHD. Multiple courses may be necessary to achieve or maintain response in some patients, and prolonged BDP therapy is a feasible alternative to prolonged systemic corticosteroids. PMID- 16041309 TI - Efficacy of reduced-intensity allogeneic stem cell transplantation in chemotherapy-refractory non-hodgkin lymphoma. AB - Chemotherapy sensitivity has been identified as an important prognostic factor in reduced-intensity allogeneic stem cell transplantation (RIST) for non-Hodgkin lymphoma (NHL). However, the effect of uniform salvage chemotherapy before RIST has not been studied prospectively. We examined whether the response to prospectively administered uniform salvage therapy (etoposide, prednisone, vincristine, cyclophosphamide, doxorubicin, and fludarabine) influenced the subsequent outcome of RIST in 28 patients with relapsed or refractory NHL. After RIST, overall survival (OS) at 36 months is 49%, whereas event-free survival (EFS) is 32%. In Cox model analyses, the response to chemotherapy was the best predictor of OS (P = .0006) and EFS (P = .0006) after RIST. Differentiating stable disease from progressive disease after salvage chemotherapy strengthened the association with survival. Among chemotherapy-sensitive patients, the median OS and EFS have not been reached. In patients with stable disease, OS and EFS at 24 months are 50% and 25%, respectively. In contrast, only 1 patient with progressive disease during salvage therapy survived longer than 12 months. These prospective data confirm the favorable prognosis for chemotherapy-sensitive NHL after RIST and suggest that chemotherapy resistance is not an absolute contraindication to RIST for NHL patients with stable disease during salvage therapy. PMID- 16041310 TI - Bone marrow transplantation for diamond-blackfan anemia. AB - Patients with Diamond-Blackfan anemia (DBA) who are unresponsive to or intolerant of corticosteroids, experience treatment failure with other treatments, develop additional cytopenias or clonal disease, or opt for curative therapy are often treated with allogeneic bone marrow transplantation. We studied the transplantation outcomes of 61 DBA patients whose data were reported to the International Bone Marrow Transplant Registry between 1984 and 2000. The median age was 7 years (range, 1-32 years). Among 55 patients with available transfusion information, 35 (64%) had received > or =20 units of blood before transplantation. Most patients (67%) received their bone marrow grafts from an HLA-matched related donor. The median time to neutrophil recovery was 17 days (range, 10-119 days) and to platelet recovery was 23 days (range, 9-119 days). Five patients did not achieve neutrophil engraftment. The 100-day mortality was 18% (95% confidence interval, 10%-29%). Grade II to IV acute graft-versus-host disease occurred in 28% (range, 17%-39%) and chronic graft-versus-host disease in 26% (range, 15%-39%). The 3-year probability of overall survival was 64% (range, 50%-74%). In univariate analysis, a Karnofsky score > or =90 and transplantation from an HLA-identical sibling donor were associated with better survival. These data suggest that allogeneic bone marrow transplantation is effective for the treatment of DBA. Transplantation before deterioration of the performance status and from an HLA-identical sibling donor may improve survival. PMID- 16041311 TI - Heterogeneous clearance of antithymocyte globulin after CD34+-selected allogeneic hematopoietic progenitor cell transplantation. AB - Antithymocyte globulins (ATG) are purified, concentrated preparations of polyclonal immunoglobulin G from hyperimmune serum of horses or rabbits immunized with human thymus lymphocytes. Both the horse and the rabbit products induce immunosuppression as a result of lymphocyte depletion and immune modulation. The exact mechanism of action is unknown but may include T-cell clearance from the circulation and modulation of T-cell activation, homing, and cytotoxic activities. Both horse and rabbit ATG include multiple antibodies against T-cell surface antigens and have been used extensively in allogeneic hematopoietic progenitor cell transplantation (HPCT) for the treatment and prevention of graft versus-host disease or graft rejection. To quantify the active ATG after HPCT, we developed a flow-based assay to measure residual ATG capable of binding to lymphocytes. In contrast to prior assays that measure total rabbit or horse immunoglobulin, this assay quantitates only the antibody capable of binding to lymphocytes, which presumably reflects the functionally active fraction of the xenoantiserum. Thirty patients with hematologic malignancies underwent T cell depleted HPCT and had ATG levels assayed during the peritransplantation period. The time required for ATG levels to decay to background was quite variable (mean, 46 days; range, 14-91 days), although most patients demonstrated a rapid early clearance followed by a slower decline. The actual mean half-life was 6.8 days (range, 2.4-14.0 days). The persistence of ATG for months after administration has significant implications for the pace of immune reconstitution after transplantation and is a potentially confounding variable in any study that involves early administration of donor lymphocyte infusions or other cellular transfer. These findings indicate that ATG levels should be explicitly measured in studies that involve early donor lymphocyte administration so that proper conclusions regarding dose, safety, and efficacy can be reached. PMID- 16041312 TI - Improved outcome for peripheral blood stem cell transplantation for advanced primary myelodysplastic syndrome. AB - Stem cell transplantation for myelodysplastic syndrome (MDS) is characterized by high transplant-related mortality (TRM), especially in older patients and those with more advanced disease. Outcome after peripheral blood stem cell transplantation (PBSCT) may be superior to earlier results with bone marrow transplantation. Forty-three patients (aged 12-73 years; median, 49 years) received an HLA-identical sibling donor PBSCT. Twenty three patients aged < or =55 years without prohibitive comorbidity received myeloablative total body irradiation-based conditioning, followed by a T cell-depleted PBSCT and delayed add-back of donor lymphocytes. Older patients or those with comorbidities (n = 20) received reduced-intensity conditioning and an unmanipulated PBSCT. Thirty seven (86%) had advanced disease (refractory anemia with excess blasts [n = 9], refractory anemia with excess blasts in transformation [n = 6], acute myelogenous leukemia [n = 13], or treatment-related MDS [n = 9]); 6 had low-risk MDS (refractory anemia or refractory anemia with ringed sideroblasts). The median follow-up was 18 months (range, 5-89 months). Actuarial probabilities of 3-year overall survival (OS), disease-free survival, relapse, and TRM were 64%, 59%, 26%, and 23%, respectively, for 34 primary MDS patients. The best results were in 19 patients younger than 50 years of age undergoing myeloablative PBSCT (actuarial probabilities of OS, disease-free survival, relapse, and TRM were 81%, 72%, 22%, and 7%, respectively). Although outcomes for all stages of primary MDS were improved, that for therapy-related MDS remained dismal, with 11% OS, because of a high relapse rate (89%). PMID- 16041313 TI - A new schedule of CHOP/rituximab plus granulocyte-macrophage colony-stimulating factor is an effective rescue for patients with aggressive lymphoma failing autologous stem cell transplantation. AB - From 1999 to 2002, 20 patients with aggressive non-Hodgkin lymphoma, among 28 who failed autologous peripheral blood progenitor cell transplantation, were rescued with cyclophosphamide, hydroxydaunomycin, Oncovin (vincristine), and prednisone (CHOP)/rituximab (RTX) and granulocyte-macrophage colony-stimulating factor (GM CSF). RTX was administered twice during each course of chemotherapy, before CHOP and after GM-CSF. This cytokine was given to increase the antibody-dependent cell mediated cytotoxicity and to reduce the leukopenia on the basis of our preliminary data, which suggested that this cytokine can upregulate CD20 expression. The relevant (World Health Organization grade 3-4) toxicity mainly consisted of myelosuppression (neutropenia in 60% of patients). Fifteen patients achieved complete remission (CR) or had a partial response, with an overall response rate of 75% (60% CR and 15% partial response). Six of the 12 patients who achieved CR relapsed: 2 died of progressive disease, 1 died of infectious complications after allogeneic transplantation, and 3 are alive in second CR. Eight patients showed progressive disease: 5 died of progressive disease, 1 of secondary acute leukemia, and 1 of infectious complications after allogeneic transplantation, whereas 1 is alive in second CR. At last follow-up, 10 patients are alive, 6 of whom are in complete continuous remission, with a median follow up of 31 months (range, 3-51 months). The projected 4-year progression-free survival is 31.4%, and the 4-year overall survival is 50%. This new association (RTX, CHOP, and GM-CSF) was feasible in approximately 70% of patients; the overall toxicity was manageable. The good response rate and the promising outcome observed in this subset of patients could be explained by the possible increased synergy between chemotherapy, RTX, and GM-CSF, which should be explored in further studies. PMID- 16041314 TI - Busulfan/melphalan/antithymocyte globulin followed by unrelated donor cord blood transplantation for treatment of infant leukemia and leukemia in young children: the Cord Blood Transplantation study (COBLT) experience. AB - A non-total body irradiation-containing preparative regimen was studied in young children (<4 years old) undergoing unrelated donor cord blood transplantation as part of the Cord Blood Transplantation trial for the treatment of acute lymphoblastic leukemia (n = 14), acute myeloid leukemia (n = 13), undifferentiated leukemia (n = 1), juvenile myelomonocytic leukemia (n = 2), and myelodysplastic syndromes (n = 2). Donor/recipient HLA matching based on low /intermediate-resolution molecular typing for HLA-A and -B and high-resolution HLA-DRB1 typing was 5/6 or 6/6 (n = 21) or 4/6 (n = 11). The preparative therapy consisted of busulfan, melphalan, and antithymocyte globulin, with cyclosporine and corticosteroids for graft-versus-host disease (GVHD) prophylaxis. The median age was 1.6 years (range, 0.5-3.9 years), and the median weight was 10.5 kg (range, 5.8-19.5 kg). Cord blood grafts contained a median of 10.7 x 10 7 nucleated cells per kilogram (range, 4.6-29.2) and 2.6 x 10(5) CD34+ cells per kilogram (range, 0.7-8.3). The cumulative incidence (CINC) of neutrophil recovery (absolute neutrophil count >500/microL) at day 42 was 0.59 (95% confidence interval [CI], 0.44-0.78) at a median of 31 days (range, 23-55 days). The CINC and Kaplan-Meier estimates of platelet engraftment at day 180 were 0.53 (95% CI, 0.34-0.69) and 0.82 (95% CI, 0.61-1.00), respectively. CINC estimates of grade III/IV acute GVHD at day 100 and chronic GVHD at 1 year were 0.25 (95% CI, 0.09 0.41) and 0.26 (95% CI, 0.09-0.44), respectively. The CINC estimate of relapse was 0.31 (95% CI, 0.16-0.47) at 2 years. With a median follow-up of 27.8 months (range, 23.4-46.7 months), the probability of survival at 1 year was 0.47 (95% CI, 0.30-0.64). A preparative regimen containing a busulfan/melphalan/antithymocyte globulin preparative regimen is well tolerated in the setting of unrelated donor cord blood transplantation for childhood leukemia and can serve as a platform preparative regimen for intensifying host immunosuppression and antileukemic therapy to allow for improved engraftment and improved relapse-free survival. PMID- 16041315 TI - Sirolimus-related toxicity in stem cell transplantation. PMID- 16041317 TI - Asymmetry on large scale: the roadmap to stereoselective processes. AB - In recent years there has been a movement in drug discovery and development away from chemical processes that produce racemic compounds towards those that produce stereochemically defined products. Asymmetric reactions are an attractive way to produce such products. Here, I discuss the factors that are important in deciding whether using an asymmetric reaction is the most appropriate approach for the large-scale synthesis of a stereochemically defined pharmaceutical compound, and highlight progress in the development of large-scale stereoselective processes. PMID- 16041318 TI - Leukotriene modifiers as potential therapeutics for cardiovascular disease. AB - Owing to their anti-inflammatory properties, leukotriene modifiers have been the primary therapeutics in asthma management for several years. Although blocking the inflammatory component of human disease is a long-standing and established concept, the use of leukotriene modifiers in treating the inflammatory component of cardiovascular disease encompassing atherosclerosis, myocardial infarction, stroke and aortic aneurysm has, surprisingly, only been seriously contemplated in the past few years. As reviewed here, several exciting studies have recently contributed to this expanding area of interest, and so far one leukotriene modifier has entered Phase II clinical trials to assess its potential for reducing the risk of heart attacks. PMID- 16041319 TI - Michelob_x is the missing inhibitor of apoptosis protein antagonist in mosquito genomes. AB - Apoptosis is implicated in the life cycle of the malaria parasite in mosquitoes. The genome project for the primary malaria vector Anopheles gambiae showed a significant expansion of the inhibitor of apoptosis protein (IAP) and caspase gene families in comparison with Drosophila. However, because of extensive sequence divergence, no orthologue was identified for the reaper/grim-like IAP antagonist genes that have a pivotal role in cell death regulation in Drosophila. Using a customized searching strategy, we identified michelob_x(mx), a gene not predicted by the genome project, as the missing IAP antagonist in the An. gambiae and other mosquito genomes. Mx has a highly conserved amino-terminal IAP-binding motif. Expression of Mx induces rapid cell death in insect cell lines and is a potent tissue ablator in vivo. Its proapoptotic activity is totally dependent on the IAP-binding motif. Like reaper in Drosophila, mx is transcriptionally induced by ultraviolet irradiation to mediate cell death. PMID- 16041321 TI - What's in a name? PMID- 16041320 TI - Hda inactivation of DnaA is the predominant mechanism preventing hyperinitiation of Escherichia coli DNA replication. AB - Initiation of DNA replication from the Escherichia coli chromosomal origin is highly regulated, assuring that replication occurs precisely once per cell cycle. Three mechanisms for regulation of replication initiation have been proposed: titration of free DnaA initiator protein by the datA locus, sequestration of newly replicated origins by SeqA protein and regulatory inactivation of DnaA (RIDA), in which active ATP-DnaA is converted to the inactive ADP-bound form. DNA microarray analyses showed that the level of initiation in rapidly growing cells that lack datA was indistinguishable from that in wild-type cells, and that the absence of SeqA protein caused only a modest increase in initiation, in agreement with flow-cytometry data. In contrast, cells lacking Hda overinitiated replication twofold, implicating RIDA as the predominant mechanism preventing extra initiation events in a cell cycle. PMID- 16041322 TI - New arrangements. PMID- 16041323 TI - Language barrier. PMID- 16041332 TI - Can a single composite resin serve all purposes? AB - The consensus view less than a decade ago was that direct posterior composites should be restricted to small restorations, preferably in premolar teeth with little, if any, occlusal function. Major advances in adhesive systems, materials and restorative techniques have combined to allow us to question this view and our increased clinical evidence base makes it appropriate to reconsider this viewpoint. PMID- 16041333 TI - Anterior dental aesthetics: dentofacial perspective. AB - The purpose of this series is to convey the principles governing our aesthetic senses. Usually meaning visual perception, aesthetics is not merely limited to the ocular apparatus. The concept of aesthetics encompasses both the time-arts such as music, theatre, literature and film, as well as space-arts such as paintings, sculpture and architecture. PMID- 16041334 TI - Salvage of an impacted canine associated with an adenomatoid odontogenic tumour: a case report. AB - The adenomatoid odontogenic tumour (AOT) has been known by a number of descriptive names (adenoameloblastoma, ameloblastic adenomatoid tumour, glandular ameloblastoma, and adenomatoid ameloblastoma) since it was first reported and later recognised as a distinct odontogenic lesion unrelated to ameloblastoma. Although it was considered to be a variant of ameloblastoma at one time leading surgeons to perform unduly aggressive surgery, the treatment outcome experience has borne out the benign, nonaggressive nature of this lesion. The AOT is now considered to be a hamartoma with completely benign behaviour. Recurrence seldom if ever occurs after surgical curettage. Thus, it appears needless to extract involved anterior teeth associated with the tumour, especially in children. We report a case in which surgical and orthodontic treatment helped to salvage an impacted mandibular canine associated with an AOT in an adolescent girl. We have not found such a procedure to have been done for this tumour in the literature. PMID- 16041335 TI - Gingival lesions as a first symptom of pemphigus vulgaris in pregnancy. AB - The erosive gingival lesions associated with vesiculobullous diseases can be an important early clinical manifestation of serious diseases such as pemphigus vulgaris (PV). PV is a vesiculobullous disease of the skin and mucosa which tends to be chronic and which normally affects people of 40-60 years of age. Its incidence varies from 0.5 to 3.2 cases per 100,000 per year. Mucosal lesions are located mainly in the oral and pharyngeal mucosa, although conjunctiva, larynx, nasal mucosa, vulva, vagina, cervix, and ano-rectal mucosa may also be involved. It is a serious mucocutaneous disease of an autoimmune nature, whose appearance during pregnancy is extremely rare. PMID- 16041341 TI - Educational needs and employment status of Scottish dental technicians. AB - OBJECTIVE: To investigate the educational needs and employment status of dental technicians in Scotland. SUBJECTS: Two hundred and fifty dental technicians with postal addresses in Scotland. DESIGN: Structured questionnaire. RESULTS: An 83% response rate was achieved following three mailings. The majority of respondents were employed in commercial dental laboratories largely within the 'central belt' of Scotland, with 96% stating they were in full-time employment. Only 33% of these essential health-care workers were voluntarily registered with the Dental Technicians' Association, suggesting that a significant number had not felt it necessary or beneficial to do so. A lack of educational structure was identified, as was poor remuneration and an absence of opportunity for career progression. Although the prospect of continuing professional development was desirable, many respondents reported that they would be penalised financially for undertaking this and, in addition, may not be given the opportunity to pursue education because of lack of co-operation from their employer. Only 47% had attended an educational event within the preceding year, and of those who had not done this, a period of two-32 years had elapsed since any CPD involvement. Of the respondents, only 34% stated that any financial assistance had been available for educational purposes, with access to education being highlighted as problematic by 68%. A total of 64% of subjects felt they were out-of-date with professional education. CONCLUSIONS: This study highlights a number of real and potential problems in the field of education in dental technology. It is apparent that change within the structure of education and professional status, although largely welcomed, may be difficult to implement. The profession, as a whole, must realise that these changes in education and employment are not optional, and should be embraced as a positive step which will hopefully raise the profile and status of dental technicians throughout the UK. PMID- 16041342 TI - Microhardness of dentine underlying ART restorations in primary molars: an in vivo pilot study. AB - OBJECTIVE: To evaluate the microhardness of dentine underlying glass ionomer restorations made with Atraumatic Restorative Treatment. DESIGN: An experimental single-centre study. SETTING: University Department, Brazil, 2001. MATERIALS AND METHODS: Sixteen children were selected showing primary molars containing carious cavities (n = 29), which were restored using Fugi IX. Subsequently, some teeth were extracted respecting Nolla stage 7 or 8 of the permanent successor, and categorised according to the post-restoration time in the mouth: G1 (baseline, immediate extraction), G2 (30 days), G3 (90 days) and G4 (180 days). Four teeth were excluded because they were not in time of normal exfoliation. The 25 extracted teeth were resin embedded, sectioned mesiodistally and prepared metallographically. Knoop microhardness analysis (Micromet 2003; 10g for 1.5 sec.) produced identations in three areas of dentine: Zone 1 (just below the restoration), Zone 3 (as close as possible to the pulp) and Zone 2 (intermediate region between 1 and 3). Non-parametric statistical tests were done--at a significance level of 5%. RESULTS: Four teeth were excluded. No significant difference was noted among the zones, inside the groups. Zones 1 and 2 showed a considerable increase of microhardness according to the time, except for Group 3 (p > 0.05). CONCLUSION: Although microhardness had increased, it was not sufficiently equal to the microhardness of the healthy dentine, regardless of the depth evaluated. PMID- 16041343 TI - Ambrosius Holbein's memento mori map for Sir Thomas More's Utopia. The meanings of a masterpiece of early sixteenth century graphic art. AB - This paper describes how, and asks why, the Renaissance artist Ambrosius Holbein hid a skull within the overall design of his woodcut map of Sir Thomas More's Utopia. (Fig. 2) This map was prepared for the 1518 Froben edition of the book, and was probably commissioned by Erasmus of Rotterdam. Its identification now is made easier by the habits of interpretation with which all dentists are equipped thanks to their skill in dental radiology, and by the recognition of teeth appearing in an unlikely disguise. PMID- 16041361 TI - Crystal structure of a bacterial homologue of Na+/Cl--dependent neurotransmitter transporters. AB - Na+/Cl--dependent transporters terminate synaptic transmission by using electrochemical gradients to drive the uptake of neurotransmitters, including the biogenic amines, from the synapse to the cytoplasm of neurons and glia. These transporters are the targets of therapeutic and illicit compounds, and their dysfunction has been implicated in multiple diseases of the nervous system. Here we present the crystal structure of a bacterial homologue of these transporters from Aquifex aeolicus, in complex with its substrate, leucine, and two sodium ions. The protein core consists of the first ten of twelve transmembrane segments, with segments 1-5 related to 6-10 by a pseudo-two-fold axis in the membrane plane. Leucine and the sodium ions are bound within the protein core, halfway across the membrane bilayer, in an occluded site devoid of water. The leucine and ion binding sites are defined by partially unwound transmembrane helices, with main-chain atoms and helix dipoles having key roles in substrate and ion binding. The structure reveals the architecture of this important class of transporter, illuminates the determinants of substrate binding and ion selectivity, and defines the external and internal gates. PMID- 16041362 TI - Architecture of floral branch systems in maize and related grasses. AB - The external appearance of flowering plants is determined to a large extent by the forms of flower-bearing branch systems, known as inflorescences, and their position in the overall structure of the plant. Branches and branching patterns are produced by tissues called shoot apical meristems. Thus, inflorescence architecture reflects meristem number, arrangement and activity, and the duration of meristem activity correlates with branch length. The inflorescences of maize, unlike those of related grasses such as rice and sorghum, predominantly lack long branches, giving rise to the tassel and familiar corncob. Here we report the isolation of the maize ramosa1 gene and show that it controls inflorescence architecture. Through its expression in a boundary domain near the nascent meristem base, ramosa1 imposes short branch identity as branch meristems are initiated. A second gene, ramosa2, acts through ramosa1 by regulating ramosa1 gene expression levels. ramosa1 encodes a transcription factor that appears to be absent in rice, is heterochronically expressed in sorghum, and may have played an important role in maize domestication and grass evolution. PMID- 16041363 TI - Potent and persistent in vivo anti-HBV activity of chemically modified siRNAs. AB - The efficacy of lipid-encapsulated, chemically modified short interfering RNA (siRNA) targeted to hepatitis B virus (HBV) was examined in an in vivo mouse model of HBV replication. Stabilized siRNA targeted to the HBV RNA was incorporated into a specialized liposome to form a stable nucleic-acid-lipid particle (SNALP) and administered by intravenous injection into mice carrying replicating HBV. The improved efficacy of siRNA-SNALP compared to unformulated siRNA correlates with a longer half-life in plasma and liver. Three daily intravenous injections of 3 mg/kg/day reduced serum HBV DNA >1.0 log(10). The reduction in HBV DNA was specific, dose-dependent and lasted for up to 7 d after dosing. Furthermore, reductions were seen in serum HBV DNA for up to 6 weeks with weekly dosing. The advances demonstrated here, including persistence of in vivo activity, use of lower doses and reduced dosing frequency are important steps in making siRNA a clinically viable therapeutic approach. PMID- 16041364 TI - In vivo imaging platform for tracking immunotherapeutic cells. AB - Cellular therapeutics show great promise for the treatment of disease, but few noninvasive techniques exist for monitoring the cells after administration. Here we present a magnetic resonance imaging (MRI) technology that uses perfluoropolyether (PFPE) agents to track cells in vivo. Fluorine MRI selectively images only the labeled cells, and a 'conventional' (1)H image places the cells in their anatomical context. We labeled phenotypically defined dendritic cells (DCs) with PFPE ex vivo and observed efficient intracellular uptake of the PFPE with little effect on DC function. We injected labeled DCs into tissue or intravenously in mice and then tracked the cells in vivo using (19)F MRI. Although we focused on DCs, which are being developed as immunotherapeutics for cancer and autoimmune diseases, this technology should be useful for monitoring a wide range of cell types in vivo. PMID- 16041365 TI - A bacterial one-hybrid system for determining the DNA-binding specificity of transcription factors. AB - The DNA-binding specificities of transcription factors can be used to computationally predict cis-regulatory modules (CRMs) that regulate gene expression. However, the absence of specificity data for the majority of transcription factors limits the widespread implementation of this approach. We have developed a bacterial one-hybrid system that provides a simple and rapid method to determine the DNA-binding specificity of a transcription factor. Using this technology, we successfully determined the DNA-binding specificity of seven previously characterized transcription factors and one novel transcription factor, the Drosophila melanogaster factor Odd-skipped. Regulatory targets of Odd skipped were successfully predicted using this information, demonstrating that the data produced by the bacterial one-hybrid system are relevant to in vivo function. PMID- 16041366 TI - High-throughput generation of small antibacterial peptides with improved activity. AB - Cationic antimicrobial peptides are able to kill a broad variety of Gram-negative and Gram positive bacteria and thus are good candidates for a new generation of antibiotics to treat multidrug-resistant bacteria. Here we describe a high throughput method to screen large numbers of peptides for improved antimicrobial activity. The method relies on peptide synthesis on a cellulose support and a Pseudomonas aeruginosa strain that constitutively expresses bacterial luciferase. A complete substitution library of 12-amino-acid peptides based on a linearized variant (RLARIVVIRVAR-NH(2)) of the bovine peptide bactenecin was screened and used to determine which substitutions at each position of the peptide chain improved activity. By combining the most favorable substitutions, we designed optimized 12-mer peptides showing broad spectrum activities with minimal inhibitory concentrations (MIC) as low as 0.5 microg/ml against Escherichia coli. Similarly, we generated an 8-mer substituted peptide that showed broad spectrum activity, with an MIC of 2 microg/ml, against E. coli and Staphylococcus aureus. PMID- 16041367 TI - Prohibitin is required for Ras-induced Raf-MEK-ERK activation and epithelial cell migration. AB - Ras proteins control the signalling pathways that are responsible for normal growth and malignant transformation. Raf protein kinases are direct Ras effector proteins that initiate the mitogen-activated protein kinase (MAPK) cascade, which mediates diverse biological functions such as cell growth, survival and differentiation. Here we show that prohibitin, a ubiquitously expressed and evolutionarily conserved protein is indispensable for the activation of the Raf MEK-ERK pathway by Ras. The membrane targeting and activation of C-Raf by Ras needs prohibitin in vivo. In addition, direct interaction with prohibitin is required for C-Raf activation. C-Raf kinase fails to interact with the active Ras induced by epidermal growth factor in the absence of prohibitin. Moreover, in prohibitin-deficient cells the adhesion complex proteins cadherin and beta catenin relocalize to the plasma membrane and thereby stabilize adherens junctions. Our data show an unexpected role of prohibitin in the activation of the Ras-Raf signalling pathway and in modulating epithelial cell adhesion and migration. PMID- 16041368 TI - Temporal and spatial control of nucleophosmin by the Ran-Crm1 complex in centrosome duplication. AB - Centrosome duplication is tightly controlled during faithful cell division, and unnecessary reduplication can lead to supernumerary centrosomes and multipolar spindles that are associated with most human cancer cells. In addition to nucleocytoplasmic transport, the Ran-Crm1 network is involved in regulating centrosome duplication to ensure the formation of a bipolar spindle. Here, we discover that nucleophosmin (NPM) may be a Ran-Crm1 substrate that controls centrosome duplication. NPM contains a functional nuclear export signal (NES) that is responsible for both its nucleocytoplasmic shuttling and its association with centrosomes, which are Ran-Crm1-dependent as they are sensitive to Crm1 specific nuclear export inhibition, either by leptomycin B (LMB) or by the expression of a Ran-binding protein, RanBP1. Notably, LMB treatment induces premature centrosome duplication in quiescent cells, which coincides with NPM dissociation from centrosomes. Moreover, deficiency of NPM by RNA interference results in supernumerary centrosomes, which can be reversed by reintroducing wild type but not NES-mutated NPM. Mutation of a potential proline-dependent kinase phosphorylation site at residue 95, from threonine to aspartic acid (T95D) within the NES motif, abolishes NPM association and inhibition of centrosome duplication. Our results are consistent with the hypothesis that the Ran-Crm1 complex may promote a local enrichment of NPM on centrosomes, thereby preventing centrosome reduplication. PMID- 16041369 TI - Multiple origins of Cajal-Retzius cells at the borders of the developing pallium. AB - Cajal-Retzius cells are critical in cortical lamination, but very little is known about their origin and development. The homeodomain transcription factor Dbx1 is expressed in restricted progenitor domains of the developing pallium: the ventral pallium (VP) and the septum. Using genetic tracing and ablation experiments in mice, we show that two subpopulations of Reelin(+) Cajal-Retzius cells are generated from Dbx1-expressing progenitors. VP- and septum-derived Reelin(+) neurons differ in their onset of appearance, migration routes, destination and expression of molecular markers. Together with reported data supporting the generation of Reelin(+) cells in the cortical hem, our results show that Cajal Retzius cells are generated at least at three focal sites at the borders of the developing pallium and are redistributed by tangential migration. Our data also strongly suggest that distinct Cajal-Retzius subtypes exist and that their presence in different territories of the developing cortex might contribute to region-specific properties. PMID- 16041370 TI - Motility-associated hair-bundle motion in mammalian outer hair cells. AB - Mammalian hearing owes its remarkable sensitivity and frequency selectivity to a local mechanical feedback process within the cochlea. Cochlear outer hair cells (OHCs) function as the key elements in the feedback loop in which the fast somatic motility of OHCs is thought to be the source of cochlear amplification. An alternative view is that amplification arises from active hair-bundle movement, similar to that seen in nonmammalian hair cells. We measured voltage evoked hair-bundle motions in the gerbil cochlea to determine if such movements were also present in mammalian OHCs. The OHCs showed bundle movement with peak responses of up to 830 nm. The movement was insensitive to manipulations that would normally block mechanotransduction in the stereocilia, and it was absent in neonatal OHCs and prestin-knockout OHCs. These findings suggest that the bundle movement originated in somatic motility and that somatic motility has a central role in cochlear amplification in mammals. PMID- 16041371 TI - Karyotypic abnormalities create discordance of germline genotype and cancer cell phenotypes. AB - The nature of mendelian inheritance assumes that all tissues in which a phenotype of interest is expressed have a uniform diploid karyotype, which is often not the case in cancer cells. Owing to nonrandom gains of chromosomes, trisomies are present in many cases of leukemia and other malignances. We used polymorphisms in the genes encoding thiopurine S-methyltransferase (TPMT), gamma-glutamyl hydrolase (GGH) and the reduced folate carrier (SLC19A1) to assess the nature of chromosomal acquisition and its influence on genotype-phenotype concordance in cancer cells. TPMT and GGH activities in somatic cells were concordant with germline genotypes, whereas activities in leukemia cells were determined by chromosomal number and whether the acquired chromosomes contained a wild-type or variant allele. Leukemia cells that had acquired an additional chromosome containing a wild-type TPMT or GGH allele had significantly lower accumulation of thioguanine nucleotides or methotrexate polyglutamates, respectively. Among these genes, there was a comparable number of acquired chromosomes with wild-type and variant alleles. Therefore, chromosomal gain can alter the concordance of germline genotype and cancer cell phenotypes, indicating that allele-specific quantitative genotyping may be required to define cancer pharmacogenomics unequivocally. PMID- 16041372 TI - Nova regulates brain-specific splicing to shape the synapse. AB - Alternative RNA splicing greatly increases proteome diversity and may thereby contribute to tissue-specific functions. We carried out genome-wide quantitative analysis of alternative splicing using a custom Affymetrix microarray to assess the role of the neuronal splicing factor Nova in the brain. We used a stringent algorithm to identify 591 exons that were differentially spliced in the brain relative to immune tissues, and 6.6% of these showed major splicing defects in the neocortex of Nova2-/- mice. We tested 49 exons with the largest predicted Nova-dependent splicing changes and validated all 49 by RT-PCR. We analyzed the encoded proteins and found that all those with defined brain functions acted in the synapse (34 of 40, including neurotransmitter receptors, cation channels, adhesion and scaffold proteins) or in axon guidance (8 of 40). Moreover, of the 35 proteins with known interaction partners, 74% (26) interact with each other. Validating a large set of Nova RNA targets has led us to identify a multi-tiered network in which Nova regulates the exon content of RNAs encoding proteins that interact in the synapse. PMID- 16041373 TI - Mutations in the endosomal ESCRTIII-complex subunit CHMP2B in frontotemporal dementia. AB - We have previously reported a large Danish pedigree with autosomal dominant frontotemporal dementia (FTD) linked to chromosome 3 (FTD3). Here we identify a mutation in CHMP2B, encoding a component of the endosomal ESCRTIII complex, and show that it results in aberrant mRNA splicing in tissue samples from affected members of this family. We also describe an additional missense mutation in an unrelated individual with FTD. Aberration in the endosomal ESCRTIII complex may result in FTD and neurodegenerative disease. PMID- 16041374 TI - A stress-sensitive reporter predicts longevity in isogenic populations of Caenorhabditis elegans. AB - When both genotype and environment are held constant, 'chance' variation in the lifespan of individuals in a population is still quite large. Using isogenic populations of the nematode Caenorhabditis elegans, we show that, on the first day of adult life, chance variation in the level of induction of a green fluorescent protein (GFP) reporter coupled to a promoter from the gene hsp-16.2 predicts as much as a fourfold variation in subsequent survival. The same reporter is also a predictor of ability to withstand a subsequent lethal thermal stress. The level of induction of GFP is not heritable, and GFP expression levels in other reporter constructs are not associated with differences in longevity. HSP-16.2 itself is probably not responsible for the observed differences in survival but instead probably reflects a hidden, heterogeneous, but now quantifiable, physiological state that dictates the ability of an organism to deal with the rigors of living. PMID- 16041375 TI - Demonstrating stratification in a European American population. AB - Population stratification occurs in case-control association studies when allele frequencies differ between cases and controls because of ancestry. Stratification may lead to false positive associations, although this issue remains controversial. Empirical studies have found little evidence of stratification in European-derived populations, but potentially significant levels of stratification could not be ruled out. We studied a European American panel discordant for height, a heritable trait that varies widely across Europe. Genotyping 178 SNPs and applying standard analytical methods yielded no evidence of stratification. But a SNP in the gene LCT that varies widely in frequency across Europe was strongly associated with height (P < 10(-6)). This apparent association was largely or completely due to stratification; rematching individuals on the basis of European ancestry greatly reduced the apparent association, and no association was observed in Polish or Scandinavian individuals. The failure of standard methods to detect this stratification indicates that new methods may be required. PMID- 16041376 TI - Heterogeneity in polymer melts from melting of polymer crystals. AB - Semi-crystalline polymers containing amorphous and crystalline regions usually have intimately mixed chains. The resulting topological constraints (entanglements) in the amorphous regions limit the drawability in the solid state. By controlled synthesis the number of entanglements can be reduced. Ultimately, crystals composed of single chains are feasible, where the chains are fully separated from each other. If such separation can be maintained in the melt a new melt state can be formed. Here we show that through slow and carefully controlled melting such polymer crystals form a heterogeneous melt with more entangled regions, where the chains are mixed, and less entangled ones, composed of individually separated chains. Chain reptation, required for the homogenization of the entanglement distribution, is found to be considerably hindered. The long-lived heterogeneous melt shows decreased melt viscosity and provides enhanced drawability on crystallization. This novel route to create heterogeneous melt should be applicable to polymers in general. PMID- 16041377 TI - Multipurpose microfluidic probe. AB - Microfluidic systems allow (bio)chemical processes to be miniaturized with the benefit of shorter time-to-result, parallelism, reduced sample consumption, laminar flow, and increased control and efficiency. However, such miniaturization inherently limits the size of the solid objects that can be processed and entails new challenges such as the interfacing of macroscopic samples with microscopic conduits. Here, we report a microfluidic probe (MFP) that overcomes these problems by combining the concepts of 'microfluidics' and of 'scanning probes'. Here, liquid boundaries formed by hydrodynamic forces underneath the MFP confine a flow of processing solution and replace the solid walls of closed microchannels. The MFP is therefore mobile and can be used to process large surfaces and objects by scanning across them. We illustrate the versatility of this concept with several examples including protein microarraying, complex gradient-formation, multiphase laminar-flow patterning, erasing, localized staining of cells and the contact-free detachment of a single cell. Many constraints imposed by the monolithic construction of microfluidic channels can now be circumvented using an MFP, opening up new avenues for microfluidic processing. PMID- 16041378 TI - 50 MHz rectifier based on an organic diode. AB - A main focus of research on organic semiconductors is their potential application in passive organic radio-frequency identification (RF-ID) tags. First prototypes working at 125 kHz have been shown by industrial research groups. However, to be commercially viable, the organic RF-ID tag would need to be compatible with the base-carrier frequency of 13.56 MHz (ref. 2). High-frequency operation has been out of reach for devices based on organic semiconducting material, because of the intrinsically low mobility of those materials. Here, we report on a rectifier based on a pentacene diode that can rectify an incoming a.c. signal at 50 MHz. At 14 MHz, a rectified voltage of 11 V for an a.c. voltage with a peak-to-peak amplitude of 36 V has been achieved. On the basis of those results, we estimate the frequency limits of an organic diode showing that even the ultra-high frequency band at around 800 MHz is within reach. PMID- 16041379 TI - Low-temperature fabrication of dye-sensitized solar cells by transfer of composite porous layers. AB - Dye-sensitized solar cells have established themselves as a potential low-cost alternative to conventional solar cells owing to their remarkably high power conversion efficiency combined with 'low-tech' fabrication processes. As a further advantage, the active layers consisting of nanoporous TiO2 are only some tens of micrometres thick and are therefore in principle suited for flexible applications. However, typical flexible plastic substrates cannot withstand the process temperatures of up to 500 degrees C commonly used for sintering the TiO2 nanoparticles together. Even though some promising routes for low-temperature sintering have been proposed, those layers cannot compete as regards electrical properties with layers obtained with the standard high-temperature process. Here we show that by a lift-off technique, presintered porous layers can be transferred to an arbitrary second substrate, and the original electrical properties of the transferred porous layers are maintained. The transfer process is greatly assisted by the application of composite layers comprising nanoparticles and nanorods. PMID- 16041380 TI - Coherent spin transport through dynamic quantum dots. AB - Spin transport and manipulation in semiconductors have been studied intensively with the ultimate goal of realizing spintronic devices. Previous work in GaAs has focused on controlling the carrier density, crystallographic orientation and dimensionality to limit the electron spin decoherence and allow transport over long distances. Here, we introduce a new method for the coherent transport of spin-polarized electronic wave packets using dynamic quantum dots (DQDs) created by the piezoelectric field of coherent acoustic phonons. Photogenerated spin carriers transported by the DQDs in undoped GaAs (001) quantum wells exhibit a spin coherence length exceeding 100 microm, which is attributed to the simultaneous control of the carrier density and the dimensionality by the DQDs during transport. In the absence of an applied magnetic field, we observe the precession of the electron spin induced by the internal magnetic field associated with the spin splitting of the conduction band (Dresselhaus term). The coherent manipulation of the precession frequency is also achieved by applying an external magnetic field. PMID- 16041381 TI - Hyperactivation of Stat3 in gp130 mutant mice promotes gastric hyperproliferation and desensitizes TGF-beta signaling. AB - The latent transcription factor Stat3 is activated by gp130, the common receptor for the interleukin (IL)-6 cytokine family and other growth factor and cytokine receptors. Ligand-induced dimerization of gp130 leads to activation of the Stat1, Stat3 and Shp2-Ras-Erk signaling pathways. Here we assess genetically the contribution of exaggerated Stat3 activation to the phenotype of gp130 (Y757F/Y757F) mice, in which a knock-in mutation disrupts the negative feedback mechanism on gp130-dependent Stat signaling. Compared to gp130 (Y757F/Y757F) mice, reduced Stat3 activation in gp130 (Y757F/Y757F) Stat3(+/-) mice increased their lifespan, prevented splenomegaly, normalized exaggerated hepatic acute phase response and lymphocyte trafficking, and suppressed the growth of spontaneously arising gastric adenomas in young mice. These lesions share histological features of gastric polyps in aging mice with monoallelic null mutations in Smad4, which encodes the common transducer for transforming growth factor (TGF)-beta signaling. Indeed, hyperactivation of Stat3 desensitizes gp130 (Y757F/Y757F) cells to the cytostatic effect of TGF-beta through transcriptional induction of inhibitory Smad7, thereby providing a novel link for cross-talk between Stat and Smad signaling in gastric homeostasis. PMID- 16041382 TI - Killed but metabolically active microbes: a new vaccine paradigm for eliciting effector T-cell responses and protective immunity. AB - We developed a new class of vaccines, based on killed but metabolically active (KBMA) bacteria, that simultaneously takes advantage of the potency of live vaccines and the safety of killed vaccines. We removed genes required for nucleotide excision repair (uvrAB), rendering microbial-based vaccines exquisitely sensitive to photochemical inactivation with psoralen and long wavelength ultraviolet light. Colony formation of the nucleotide excision repair mutants was blocked by infrequent, randomly distributed psoralen crosslinks, but the bacterial population was able to express its genes, synthesize and secrete proteins. Using the intracellular pathogen Listeria monocytogenes as a model platform, recombinant psoralen-inactivated Lm DeltauvrAB vaccines induced potent CD4(+) and CD8(+) T-cell responses and protected mice against virus challenge in an infectious disease model and provided therapeutic benefit in a mouse cancer model. Microbial KBMA vaccines used either as a recombinant vaccine platform or as a modified form of the pathogen itself may have broad use for the treatment of infectious disease and cancer. PMID- 16041383 TI - Angiotensin II and EGF receptor cross-talk in chronic kidney diseases: a new therapeutic approach. AB - Mechanisms of progression of chronic renal diseases, a major healthcare burden, are poorly understood. Angiotensin II (AngII), the major renin-angiotensin system effector, is known to be involved in renal deterioration, but the molecular pathways are still unknown. Here, we show that mice overexpressing a dominant negative isoform of epidermal growth factor receptor (EGFR) were protected from renal lesions during chronic AngII infusion. Transforming growth factor-alpha (TGF-alpha) and its sheddase, TACE (also known as ADAM17), were induced by AngII treatment, TACE was redistributed to apical membranes and EGFR was phosphorylated. AngII-induced lesions were substantially reduced in mice lacking TGF-alpha or in mice given a specific TACE inhibitor. Pharmacologic inhibition of AngII prevented TGF-alpha and TACE accumulation as well as renal lesions after nephron reduction. These findings indicate a crucial role for AngII-dependent EGFR transactivation in renal deterioration and identify in TACE inhibitors a new therapeutic strategy for preventing progression of chronic renal diseases. PMID- 16041384 TI - NFAT and Osterix cooperatively regulate bone formation. AB - Immunosuppressants are crucial in the prevention of detrimental immune reactions associated with allogenic organ transplantation, but they often cause adverse effects in a number of biological systems, including the skeletal system. Calcineurin inhibitors FK506 and cyclosporin A inhibit nuclear factor of activated T cells (NFAT) activity and induce strong immunosuppression. Among NFAT proteins, NFATc1 is crucial for the differentiation of bone-resorbing osteoclasts. Here we show FK506 administration induces the reduction of bone mass despite a blockade of osteoclast differentiation. This reduction is caused by severe impairment of bone formation, suggesting that NFAT transcription factors also have an important role in the transcriptional program of osteoblasts. In fact, bone formation is inhibited in Nfatc1- and Nfatc2-deficient cells as well as in FK506-treated osteoblasts. Overexpression of NFATc1 stimulates Osterix dependent activation of the Col1a1 (encoding type I collagen) promoter, but not Runx2-dependent activation of the Bglap1 (encoding osteocalcin) promoter. NFAT and Osterix form a complex that binds to DNA, and this interaction is important for the transcriptional activity of Osterix. Thus, NFAT and Osterix cooperatively control osteoblastic bone formation. These results may provide important insight into the management of post-transplantation osteoporosis as well as a new strategy for promoting bone regeneration in osteopenic disease. PMID- 16041385 TI - Molecular architecture of a eukaryotic DNA transposase. AB - Mobile elements and their inactive remnants account for large proportions of most eukaryotic genomes, where they have had central roles in genome evolution. Over 50 years ago, McClintock reported a form of stress-induced genome instability in maize in which discrete DNA segments move between chromosomal locations. Our current mechanistic understanding of enzymes catalyzing transposition is largely limited to prokaryotic transposases. The Hermes transposon from the housefly is part of the eukaryotic hAT superfamily that includes hobo from Drosophila, McClintock's maize Activator and Tam3 from snapdragon. We report here the three dimensional structure of a functionally active form of the transposase from Hermes at 2.1-A resolution. The Hermes protein has some structural features of prokaryotic transposases, including a domain with a retroviral integrase fold. However, this domain is disrupted by the insertion of an additional domain. Finally, transposition is observed only when Hermes assembles into a hexamer. PMID- 16041386 TI - The HIV-1 capsid protein C-terminal domain in complex with a virus assembly inhibitor. AB - Immature HIV particles bud from infected cells after assembly at the cytoplasmic side of cellular membranes. This assembly is driven by interactions between Gag polyproteins. Mature particles, each containing a characteristic conical core, are later generated by proteolytic maturation of Gag in the virion. The C terminal domain of the HIV-1 capsid protein (C-CA) has been shown to contain oligomerization determinants essential for particle assembly. Here we report the 1.7-A-resolution crystal structure of C-CA in complex with a peptide capable of inhibiting immature- and mature-like particle assembly in vitro. The peptide inserts as an amphipathic alpha-helix into a conserved hydrophobic groove of C CA, resulting in formation of a compact five-helix bundle with altered dimeric interactions. This structure thus reveals the details of an allosteric site in the HIV capsid protein that can be targeted for antiviral therapy. PMID- 16041387 TI - A peptide inhibitor of HIV-1 assembly in vitro. AB - Formation of infectious HIV-1 involves assembly of Gag polyproteins into immature particles and subsequent assembly of mature capsids after proteolytic disassembly of the Gag shell. We report a 12-mer peptide, capsid assembly inhibitor (CAI), that binds the capsid (CA) domain of Gag and inhibits assembly of immature- and mature-like capsid particles in vitro. CAI was identified by phage display screening among a group of peptides with similar sequences that bind to a single reactive site in CA. Its binding site was mapped to CA residues 169-191, with an additional contribution from the last helix of CA. This result was confirmed by a separate X-ray structure analysis showing that CAI inserts into a conserved hydrophobic groove and alters the CA dimer interface. The CAI binding site is a new target for antiviral development, and CAI is the first known inhibitor directed against assembly of immature HIV-1. PMID- 16041388 TI - Target RNA motif and target mRNAs of the Quaking STAR protein. AB - Quaking viable (Qk(v)) mice have developmental defects that result in their characteristic tremor. The quaking (Qk) locus expresses alternatively spliced RNA binding proteins belonging to the STAR family. To characterize the RNA binding specificity of the QKI proteins, we selected for RNA species that bound QKI from random pools of RNAs and defined the QKI response element (QRE) as a bipartite consensus sequence NACUAAY-N(1-20)-UAAY. A bioinformatic analysis using the QRE identified the three known RNA targets of QKI and 1,430 new putative mRNA targets, of which 23 were validated in vivo. A large proportion of the mRNAs are implicated in development and cell differentiation, as predicted from the phenotype of the Qk(v) mice. In addition, 24% are implicated in cell growth and/or maintenance, suggesting a role for QKI in cancer. PMID- 16041389 TI - An essential function for the calcium-promoted Ras inactivator in Fcgamma receptor-mediated phagocytosis. AB - Fc receptor (FcR)-mediated phagocytosis requires activation of the Rho GTPases Cdc42 and Rac1, but how they are recruited to the FcR is unknown. Here we show that the calcium-promoted Ras inactivator (CAPRI), a Ras GTPase-activating protein, functions as an adaptor for Cdc42 and Rac1 during FcR-mediated phagocytosis. CAPRI-deficient macrophages had impaired FcgammaR-mediated phagocytosis and oxidative burst, as well as defective activation of Cdc42 and Rac1. CAPRI interacted constitutively with both Cdc42 and Rac1 and translocated to phagocytic cups during FcgammaR-mediated phagocytosis. CAPRI-deficient mice had an impaired innate immune response to bacterial infection. These results suggest that CAPRI provides a link between FcgammaR and Cdc42 and Rac1 and is essential for innate immune responses. PMID- 16041391 TI - The need for education in pharmacogenomics: a regulatory perspective. PMID- 16041392 TI - Genetic factors influencing pyrimidine-antagonist chemotherapy. AB - Pyrimidine antagonists, for example, 5-fluorouracil (5-FU), cytarabine (ara-C) and gemcitabine (dFdC), are widely used in chemotherapy regimes for colorectal, breast, head and neck, non-small-cell lung cancer, pancreatic cancer and leukaemias. Extensive metabolism is a prerequisite for conversion of these pyrimidine prodrugs into active compounds. Interindividual variation in the activity of metabolising enzymes can affect the extent of prodrug activation and, as a result, act on the efficacy of chemotherapy treatment. Genetic factors at least partly explain interindividual variation in antitumour efficacy and toxicity of pyrimidine antagonists. In this review, proteins relevant for the efficacy and toxicity of pyrimidine antagonists will be summarised. In addition, the role of germline polymorphisms, tumour-specific somatic mutations and protein expression levels in the metabolic pathways and clinical pharmacology of these drugs are described. Germline polymorphisms of uridine monophosphate kinase (UMPK), orotate phosphoribosyl transferase (OPRT), thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD) and methylene tetrahydrofolate reductase (MTHFR) and gene expression levels of OPRT, UMPK, TS, DPD, uridine phosphorylase, uridine kinase, thymidine phosphorylase, thymidine kinase, deoxyuridine triphosphate nucleotide hydrolase are discussed in relation to 5-FU efficacy. Cytidine deaminase (CDD) and 5'-nucleotidase (5NT) gene polymorphisms and CDD, 5NT, deoxycytidine kinase and MRP5 gene expression levels and their potential relation to dFdC and ara-C cytotoxicity are reviewed. PMID- 16041393 TI - Only low levels of exogenous N-acetyltransferase can be achieved in transgenic mice. AB - Therapeutic and environmental aromatic amines and hydrazines are substrates for the arylamine N-acetyltransferases (NAT). In all, 10 transgenic lines containing either the human NAT1 or NAT2 transgene were developed using multiple promoters. The presence of the transgene was confirmed by determining copy number, mRNA and enzyme activity. Despite some lines having high copy numbers of the transgene, only modest or no increases in enzymatic activity could be found in a variety of tissues. The NAT1 transgene could not be bred to homozygosity. The cytomegalovirus (CMV)-promoted NAT1 transgene increased endogenous Nat1 mRNA levels in liver and had little effect on endogenous Nat2 mRNA levels. The presence of the CMV-promoted NAT2 transgene appeared to suppress endogenous hepatic Nat2 mRNA, but did not alter Nat1 mRNA levels. The failure to achieve high expression of any of the transgenes suggests that overexpression of NAT genes may have harmful effects during development. PMID- 16041395 TI - Interactions of metoclopramide and ergotamine with human 5-HT(3A) receptors and human 5-HT reuptake carriers. AB - The actions of metoclopramide and ergotamine, drugs which are used as a combined migraine medication, on human (h)5-HT3A receptors and 5-HT reuptake carriers, stably expressed in HEK-293 cells, were studied with patch-clamp- and ([3H]5-HT) uptake techniques. At clinical concentrations, metoclopramide inhibited peak and integrated currents through h5-HT3A receptors concentration-dependently (IC50 = 0.064 and 0.076 microM, respectively) when it was applied in equilibrium (60 s before and during 5-HT (30 microM) exposure). The onset and offset time constants of metoclopramide action were 1.3 and 2.1 s, respectively. The potency of metoclopramide when exclusively applied during the agonist pulse decreased more than 200-fold (IC50 = 19.0 microM, peak current suppression). Metoclopramide (0.10 microM) did not alter the EC50 of 5-HT-induced peak currents. In contrast to the lack of competitive interaction between metoclopramide and 5-HT in this functional assay, metoclopramide inhibited specific [3H]GR65630 binding to human h5-HT3A receptors in a surmountable manner. This seeming discrepancy between functional studies and radioligand binding experiments may be accounted for by (1) the slow kinetics of inhibition of peak currents by metoclopramide compared with the fast onset and offset kinetics of 5-HT-induced currents and (2) the low efficacy of metoclopramide in inhibiting radioligand binding (e.g. only 20% binding inhibition compared to 79% peak current suppression by 200 nM metoclopramide). At low concentrations (1-10 nM), ergotamine had no effect on 5 HT (30 microM)-induced peak currents. Above clinical concentrations, ergotamine (>3 microM) inhibited them. When both drugs were applied together (0.10 microM metoclopramide +0.001 to 0.01 microM ergotamine), an inhibition of both, peak and integrated current responses was observed. Neither metoclopramide (< or =30 microM) nor ergotamine (< or =30 microM) had an effect on the 5-HT reuptake carrier as they did not alter the citalopram-sensitive [3H]5-HT uptake. PMID- 16041396 TI - Intrinsic sensory deprivation induced by neonatal capsaicin treatment induces changes in rat brain and behaviour of possible relevance to schizophrenia. AB - Schizophrenia is considered to be a neurodevelopmental disorder with origins in the prenatal or neonatal period. Brains from subjects with schizophrenia have enlarged ventricles, reduced cortical thickness (CT) and increased neuronal density in the prefrontal cortex compared with those from normal subjects. Subjects with schizophrenia have reduced pain sensitivity and niacin skin flare responses, suggesting that capsaicin-sensitive primary afferent neurons might be abnormal in schizophrenia. This study tested the hypothesis that intrinsic somatosensory deprivation, induced by neonatal capsaicin treatment, causes changes in the brains of rats similar to those found in schizophrenia. Wistar rats were treated with capsaicin, 50 mg kg(-1) subcutaneously, or vehicle (control) at 24-36 h of life. At 5-7 weeks behavioural observations were made, and brains removed, fixed and sectioned. The mean body weight of capsaicin treated rats was not significantly different from control, but the mean brain weight of male, but not female, rats, was significantly lower than control. Capsaicin-treated rats were hyperactive compared with controls. The hyperactivity was abolished by haloperidol. Coronal brain sections of capsaicin-treated rats had smaller cross-sectional areas, reduced CT, larger ventricles and aqueduct, smaller hippocampal area and reduced corpus callosum thickness, than brain sections from control rats. Neuronal density was increased in several cortical areas and the caudate putamen, but not in the visual cortex. It is concluded that neonatal capsaicin treatment of rats produces brain changes that are similar to those found in brains of subjects with schizophrenia. PMID- 16041397 TI - Endogenous opioid mechanisms partially mediate P2X3/P2X2/3-related antinociception in rat models of inflammatory and chemogenic pain but not neuropathic pain. AB - P2X3/P2X2/3 receptors have emerged as important components of nociception. However, there is limited information regarding the neurochemical systems that are affected by antagonism of the P2X3/P2X2/3 receptor and that ultimately contribute to the ensuing antinociception. In order to determine if the endogenous opioid system is involved in this antinociception, naloxone was administered just prior to the injection of a selective P2X3/P2X2/3 receptor antagonist, A-317491, in rat models of neuropathic, chemogenic, and inflammatory pain. Naloxone (1-10 mg kg(-1), i.p.), dose-dependently reduced the antinociceptive effects of A-317491 (1-300 micromol kg(-1), s.c.) in the CFA model of thermal hyperalgesia and the formalin model of chemogenic pain (2nd phase), but not in the L5-L6 spinal nerve ligation model of neuropathic allodynia. In comparison experiments, the same doses of naloxone blocked or attenuated the actions of morphine (2 or 8 mg kg(-1), s.c.) in each of these behavioral models. Injection of a peripheral opioid antagonist, naloxone methiodide (10 mg kg(-1), i.p.), did not affect A-317491-induced antinociception in the CFA and formalin assays, suggesting that the opioid component of this antinociception occurred within the CNS. Furthermore, this utilization of the central opioid system could be initiated by antagonism of spinal P2X3/P2X2/3 receptors since the antinociceptive actions of intrathecally delivered A-317491 (30 nmol) in the formalin model were reduced by both intrathecally (10-50 nmol) and systemically (10 mg kg(-1), i.p.) administered naloxone. This utilization of the opioid system was not specific to A-317491 since suramin-, a nonselective P2X receptor antagonist, induced antinociception was also attenuated by naloxone. In in vitro studies, A-317491 (3-100 microM) did not produce any agonist response at delta opioid receptors expressed in NG108-15 cells. A-317491 had been previously shown to be inactive at the kappa and mu opioid receptors. Furthermore, naloxone, at concentrations up to 1 mM, did not compete for [3H] A-317491 binding in 1321N1 cells expressing human P2X3 receptors. Taken together, these results indicate that antagonism of spinal P2X3/P2X2/3 receptors results in an indirect activation of the opioid system to alleviate inflammatory hyperalgesia and chemogenic nociception. PMID- 16041398 TI - A comparison of the effects of unfractionated heparin, dalteparin and danaparoid on vascular endothelial growth factor-induced tumour angiogenesis and heparanase activity. AB - Disseminated intravascular coagulation (DIC) is the most common complication of solid tumours. In this study, the effectiveness of three polysaccharide anticoagulants (PSAs), at therapeutic doses, at inhibiting solid tumour growth was investigated. Mice with tumour xenografts were subcutaneously injected with either unfractionated heparin (UFH; 200 units kg(-1) day(-1)), dalteparin (75 units kg(-1) day(-1)) or danaparoid (50 units kg(-1) day(-1)). At these concentrations, these PSAs are equieffective at inhibiting blood coagulation activated factor X. In mice with Lewis lung carcinoma (LLC) tumours dalteparin and, to a lesser extent, UFH inhibited both tumour growth and angiogenesis, whereas danaparoid did not. In contrast, in mice with KLN205 tumours, all the PSAs inhibited tumour growth and angiogenesis. All the PSAs significantly inhibited proliferation, migration of endothelial cells and vessel formation in matrigel plugs containing vascular endothelial growth factor (VEGF) and there were no significant differences between these effects of the PSAs. The PSAs had no effect on endothelial cell tubular formation in vitro. Although all the PSAs inhibited VEGF production in KLN205 tumours in vivo and cells in vitro, in LLC tumours and cells only UFH and dalteparin inhibited VEGF production, whereas danaparoid did not. In both LLC and KLN205 tumours in vivo, heparanase activity was inhibited by UFH and dalteparin, but not by danaparoid. Hence, UFH and dalteparin may be more effective than danaparoid at inhibiting cancer progression in DIC patients with solid tumours, due at least in part to their ability to suppress VEGF and heparanase in tumours. PMID- 16041399 TI - Ethyl caffeate suppresses NF-kappaB activation and its downstream inflammatory mediators, iNOS, COX-2, and PGE2 in vitro or in mouse skin. AB - Ethyl caffeate, a natural phenolic compound, was isolated from Bidens pilosa, a medicinal plant popularly used for treating certain inflammatory syndromes. The purpose of this study was to investigate the structural activity, and the anti inflammatory functions and mechanism(s) of ethyl caffeate. Ethyl caffeate was found to markedly suppress the lipopolysaccharide (LPS)-induced nitric oxide (NO) production (IC(50) = 5.5 microg ml(-1)), mRNA and protein expressions of inducible nitric oxide synthase (iNOS), and prostaglandin E(2) (PGE(2)) production in RAW 264.7 macrophages. Transient gene expression assays using human cox-2 promoter construct revealed that ethyl caffeate exerted an inhibitory effect on cox-2 transcriptional activity in 12-O-tetradecanoylphorbol-13-acetate (TPA)-treated MCF-7 cells. Immunohistochemical studies of mouse skin demonstrated that TPA-induced COX-2 expression was significantly inhibited by ethyl caffeate with a superior effect to that of celecoxib, a nonsteroidal anti-inflammatory drug. The phosphorylation and degradation of inhibitor kappaB (IkappaB) and the translocation of nuclear transcription factor-kappaB (NF-kappaB) into the nucleus, as well as the activation of mitogen-activated protein kinases (MAPKs) induced by LPS in macrophages, were not affected by ethyl caffeate. Ethyl caffeate, however, could inhibit NF-kappaB activation by impairing the binding of NF-kappaB to its cis-acting element. These results suggest that ethyl caffeate suppresses iNOS and COX-2 expressions partly through the inhibition of the NF kappaB.DNA complex formation. Structure-activity relationship analyses suggested that the catechol moiety and alpha,beta-unsaturated ester group in ethyl caffeate are important and essential structural features for preventing NF-kappaB.DNA complex formation. This study provides an insight into the probable mechanism(s) underlying the anti-inflammatory and therapeutic properties of ethyl caffeate. PMID- 16041400 TI - Comparison of the biological activities of anagrelide and its major metabolites in haematopoietic cell cultures. AB - The platelet-lowering drug anagrelide inhibits bone marrow megakaryocytopoiesis by an unknown mechanism. Recently, it was found that anagrelide is bio transformed in humans into two major metabolites (6,7-dichloro-3-hydroxy-1,5 dihydro-imidazo[2,1-b]quinazolin-2-one (BCH24426) and 2-amino-5,6-dichloro-3,4, dihydroquinazoline (RL603). Whether these metabolites have biological activities that may underlie the mode of action of the parent drug is presently unclear. To clarify this question here we have compared the activities of anagrelide, BCH24426 and RL603 on the growth and differentiation of CD34(+) haematopoietic progenitor cells in liquid culture and on the migration of differentiated megakaryocytes. Incubation with either anagrelide, BCH24426 or RL603 did not affect the early expansion of CD34(+) cells stimulated by thrombopoietin. In contrast, both anagrelide and BCH24426 potently inhibited the development of megakaryocytes (IC(50) +/- s.e.m. = 26 +/- 4 and 44 +/- 6 nM, respectively), whereas RL603 showed no significant effect. Anagrelide and BCH24426 did not affect erythroid or myelomonocytic differentiation stimulated by erythropoietin or granulocyte-macrophage colony-stimulating factor, demonstrating the selectivity of these compounds against the megakaryocytic lineage. Neither anagrelide nor its metabolites showed a significant effect on the migratory response of megakaryocytes towards stromal cell-derived factor-1alpha. Although BCH24426 was shown to be considerably more potent than anagrelide as an inhibitor of phosphodiesterase type III (PDEIII) (IC(50) = 0.9 vs 36 nM) this activity did not correlate with the potency of inhibition of megakaryocyte development. Furthermore, other PDEIII inhibitors of widely differing potency were shown to have negligible effects on megakaryocytopoiesis. Taken together our results demonstrate that anagrelide and BCH24426 target a cellular event involved specifically in the megakaryocyte differentiation programme, which is independent of PDEIII inhibition. PMID- 16041401 TI - Effect of clopidogrel on the expression of inflammatory markers in rabbit ischemic coronary artery. AB - Inflammation and platelet activation are critical phenomena in the setting of acute coronary syndromes. Platelets may contribute to increase ischemic injury by enhancing the inflammatory response of leukocytes and endothelial myocardial cells. Pharmacological inhibition of platelet activation prevents ischemic complications in patients with coronary diseases. Agents directed against the integrin glycoprotein IIb/IIIa (GP IIb/IIIa) receptor not only inhibit platelet aggregation but also have been demonstrated to limit the inflammatory response in acute coronary syndromes. The question then raised is if the inhibition of platelet activation by other mechanisms than the blockade of GP IIb/IIIa may also exert anti-inflammatory effects. The aim of the present study was to analyze if clopidogrel may exert anti-inflammatory effects during the acute phase of myocardial infarction. A ligature was placed around the left anterior descending coronary artery of New Zealand White rabbits. After 15 min of ischemia, the myocardium was reperfused and the ischemic coronary artery was isolated 24 h after the ischemia. A group of ischemic rabbits was given a single oral dose of clopidogrel (20 mg kg(-1)) just after the arterial occlusion and the animal was recovered. Sham-operated animals served as control. P-selectin expression was significantly increased in infarcted rabbits with respect to control rabbits. Clopidogrel administration reduced P-selectin expression with respect to untreated infarcted rabbits. CD40 ligand and tissue factor expression was increased in the ischemic coronary artery and reduced after clopidogrel administration. Clopidogrel also protected endothelial nitric oxide synthase protein expression in the ischemic coronary artery, a protein that has been found downregulated under inflammatory conditions. In conclusion, inhibition of platelet activation by clopidogrel exerted anti-inflammatory effects on the ischemic coronary artery. PMID- 16041402 TI - Novel polyisoprenyl phosphates block phospholipase D and human neutrophil activation in vitro and murine peritoneal inflammation in vivo. AB - Leukocyte production of reactive oxygen species (ROS) is an essential component of the antimicrobial armament mounted during host defense, but when released to the extracellular milieu ROS can also injure host tissues and provoke inflammation. Polyisoprenyl phosphates (PIPPs) are constituents of human leukocyte membranes that regulate pivotal intracellular enzymes, such as phospholipase D (PLD). We prepared new PIPP mimetics and studied their impact in vivo on leukocyte activation, including ROS generation, in acute inflammation. In a stereospecific and concentration-dependent manner, the PIPP mimetics directly regulated Streptomyces chromofuscus phospholipase D (sPLD) action. The IC(50) for a (Z)-isomer of endogenous presqualene diphosphate (PSDP) was 100 nM. Structure activity relationships were also determined for PIPP mimetic inhibition of recombinant human PLD1b, a prominent isoform in human leukocytes. The PIPP mimetic rank order for PLD1b inhibition differed from sPLD, although the (Z)-PSDP isomer remained the most potent PIPP mimetic for inhibition of both enzymes. Truncation of PLD1b to its catalytic core uncovered potential regulatory roles for both PSDP's isoprenoid and diphosphate moieties. The (Z)-PSDP isomer reduced ROS production by activated human leukocytes and decreased murine neutrophil accumulation (65.6%) and ROS production (38.5%) in vivo during zymosan A initiated peritonitis. When administered intraperitoneally 2 h after zymosan A, the (Z)-PSDP isomer decreased in vivo neutrophil accumulation (72.5%) and ROS generation (74.4%) 6 h later in peritoneal exudates. Together, these results provide new means to protect and control unchecked inflammatory responses that characterize many human diseases. PMID- 16041403 TI - Developing skeletal muscle cells express functional muscarinic acetylcholine receptors coupled to different intracellular signaling systems. AB - This study analyzed the expression of muscarinic acetylcholine receptors (mAChRs) in the rat cultured skeletal muscle cells and their coupling to G protein, phospholipase C and adenylyl cyclase (AC). Our results showed the presence of a homogeneous population of [(3)H]methyl-quinuclidinyl benzilate-binding sites in the membrane fraction from the rat cultured muscle (K(D) = 0.4 nM, B(max) = 8.9 fmol mg protein(-1)). Specific muscarinic binding sites were also detected in denervated diaphragm muscles from adult rats and in myoblasts isolated from newborn rats. Activation of mAChRs with carbachol induced specific [(35)S]GTPgammaS binding to cultured muscle membranes and potentiated the forskolin-dependent stimulation of AC. These effects were totally inhibited by 0.1-1 microM atropine. In addition, mAChRs were able to stimulate generation of diacylglycerol (DAG) in response to acetylcholine, carbachol or selective mAChR agonist oxotremorine-M. The carbachol-dependent increase in DAG was inhibited in a concentration-dependent manner by mAChR antagonists atropine, pirenzepine and 4 DAMP mustard. Finally, activation of these receptors was correlated with increased synthesis of acetylcholinesterase, via a PKC-dependent pathway. Taken together, these results indicate that expression of mAChRs, coupled to G protein and distinct intracellular signaling systems, is a characteristic of noninnervated skeletal muscle cells and may be responsible for trophic influences of acetylcholine during formation of the neuromuscular synapse. PMID- 16041407 TI - Enzymatic function of hemoglobin as a nitrite reductase that produces NO under allosteric control. AB - Hypoxic vasodilation is a fundamental, highly conserved physiological response that requires oxygen and/or pH sensing coupled to vasodilation. While this process was first characterized more than 80 years ago, the precise identity and mechanism of the oxygen sensor and mediators of vasodilation remain uncertain. In support of a possible role for hemoglobin (Hb) as a sensor and effector of hypoxic vasodilation, here we show biochemical evidence that Hb exhibits enzymatic behavior as a nitrite reductase, with maximal NO generation rates occurring near the oxy-to-deoxy (R-to-T) allosteric structural transition of the protein. The observed rate of nitrite reduction by Hb deviates from second-order kinetics, and sigmoidal reaction progress is determined by a balance between 2 opposing chemistries of the heme in the R (oxygenated conformation) and T (deoxygenated conformation) allosteric quaternary structures of the Hb tetramer- the greater reductive potential of deoxyheme in the R state tetramer and the number of unligated deoxyheme sites necessary for nitrite binding, which are more plentiful in the T state tetramer. These opposing chemistries result in a maximal nitrite reduction rate when Hb is 40-60% saturated with oxygen (near the Hb P50), an apparent ideal set point for hypoxia-responsive NO generation. These data suggest that the oxygen sensor for hypoxic vasodilation is determined by Hb oxygen saturation and quaternary structure and that the nitrite reductase activity of Hb generates NO gas under allosteric and pH control. PMID- 16041408 TI - Autoantigen, innate immunity, and T cells cooperate to break B cell tolerance during bacterial infection. AB - Autoantibody production during infections is considered to result from nonspecific activation of low-affinity autoreactive B cells. Whether this can lead to autoimmune disease remains uncertain. We show that chronic infection by Borrelia burgdorferi of Tg animals expressing human rheumatoid factor (RF) B cells (of low or intermediate affinities) in the absence or in the constitutive presence of the autoantigen (represented here by chimeric IgG with human constant region) breaks their state of immunological ignorance, leading to the production of RFs. Surprisingly, this production was more pronounced in intermediate affinity RF Tg mice co-expressing the autoantigen. This overproduction was mediated by immune complexes and involved synergistic signaling between the B cell receptor and Toll-like receptors and T cell help. These findings indicate that chronic infection can activate autoreactive B cells with significant affinity and creates conditions that can drive them to differentiate into memory cells. Such cells may have some physiological yet undetermined role, but in autoimmune-prone individuals, this scenario may initiate autoimmunity. PMID- 16041409 TI - Saturated fat-rich diet enhances selective uptake of LDL cholesteryl esters in the arterial wall. AB - Plasma LDL levels and atherosclerosis both increase on a saturated fat-rich (SAT) diet. LDL cholesterol delivery to tissue may occur via uptake of the LDL particles or via selective uptake (SU), wherein cholesteryl ester (CE) enters cells without concomitant whole-particle uptake. It is not known how dietary fats might directly affect arterial LDL-CE uptake and whether SU is involved. Thus, mice that are relatively atherosclerosis resistant (C57BL/6) or susceptible to atherosclerosis (apoE) were fed a chow or SAT diet and injected with double radiolabeled or fluorescent-labeled human LDL to independently trace LDL-CE core and whole-particle uptake, respectively. Our results show that a SAT diet increased contributions of SU to total arterial LDL-CE delivery in C57BL/6 and apoE mice. The SAT diet increased plasma fatty acid and cholesterol levels; cholesterol, but not fatty acid, levels correlated with SU, as did the degree of atherosclerosis. Increased SU did not correlate with arterial scavenger receptor class B type I levels but paralleled increased lipoprotein lipase (LPL) levels and LPL distribution in the arterial wall. These studies suggest that arterial LDL-CE delivery via SU can be an important mechanism in vivo and that dietary influences on arterial LPL levels and atherogenesis modulate arterial LDL-CE delivery, cholesterol deposition, and SU. PMID- 16041410 TI - Induction of mucosal tolerance in Peyer's patch-deficient, ligated small bowel loops. AB - To explore the requirement for M cells and the Peyer's patch (PP) in induction of oral tolerance and address the potential in vivo role of intestinal epithelial cells as nonprofessional APCs, we have attempted to induce tolerance in mice with ligated small bowel loops without M cells and Peyer's patches. A 2-centimeter section of vascularized small bowel was spliced away from the gut without disruption of the mesenteric attachments. We introduced OVA directly into the lumen of the loop prior to footpad immunization. By excising segments of bowel that contain PPs in some mice and segments without patches in others, we could study the necessity of the M cell and the underlying patch versus epithelial cells in induction of mucosal tolerance. We show that OVA-specific T cell proliferation and serum antibody responses are reduced in mice that have previously been given OVA both in PP-containing loops and in loops without patches. Furthermore, both high- and low-dose tolerance could be induced in the absence of PPs. Low-dose tolerance is associated with bystander suppression and requires IL-10, which indicates active suppression and the induction of regulatory cells. These data suggest that there is a critical role for components of the mucosal immune system other than PPs in antigen sampling and induction of oral tolerance. PMID- 16041411 TI - Amidopyrroles: from anion receptors to membrane transport agents. AB - Amidopyrroles have been employed in a variety of anion receptors and sensors. 2,5 Bisamidopyrroles show selective oxo-anion complexation properties in organic solution whilst bis-amides containing dipyrrolylmethane groups form strong complexes with dihydrogen phosphate anions in mixtures of DMSO-d6 and water. Deprotonation of the 2,5-bisamidopyrrole unit can lead to interesting solid-state structures including the formation of orthogonal hydrogen bonded dimers. Amidopyrrole groups have also been employed in receptors for ion-pairs and in membrane transport agents for HCl. PMID- 16041412 TI - Co-transport of H+/Cl- by a synthetic prodigiosin mimic. AB - An amidopyrrole with appended imidazole group can bind and co-transport H+/Cl- across vesicle membranes much more effectively than an analogue with an appended pyridyl group. PMID- 16041413 TI - Low power upconversion using MLCT sensitizers. AB - Selective low energy excitation of the metal-to-ligand charge transfer (MLCT) transition in [Ru(dmb)(3)](2+)(dmb = 4,4'-dimethyl-2,2'-bipyridine) in the presence of anthracene or 9,10-diphenylanthracene yields easily visualized upconverted singlet fluorescence resulting from triplet-triplet annihilation at low excitation power. PMID- 16041414 TI - On the CH...Cu agostic interaction: chiral copper(II) compounds with ephedrine and pseudoephedrine derivatives. AB - The ephedrine derivative, (H2ceph), yields [Cu(Hceph)2], showing a CH...Cu(II) agostic interaction; while in the analogous compound [Cu(Hcpse)2], with pseudoephedrine (H2cpse), that interaction is absent, despite the fact that these two diasteromers differ only in the orientation of the methyl and phenyl groups: erythro in H2ceph and threo in H2cpse. The X-ray crystal structure of [Cu(Hceph)2], indicates a Cu...HC length of 2.454 A and the theoretical study reveals the formation of a Cu...HC bond since the associated electronic density shows both a bond critical point and a bond ring critical point. PMID- 16041415 TI - Enlarged pi-electronic network of a meso-meso, beta-beta, beta-beta triply linked dibenzoporphyrin dimer that exhibits a large two-photon absorption cross section. AB - Enlargement of the pi-electronic network of meso-meso, beta-beta, beta-beta triply linked diporphyrin has been exploited by preparing a corresponding dibenzo fused porphyrin dimer that exhibits a perturbed absorption spectrum and a large two-photon absorption cross section. PMID- 16041416 TI - A surfactant-encapsulated polyoxometalate complex towards a thermotropic liquid crystal. AB - A novel surfactant-encapsulated terbium-substituted heteropolyoxotungstate complex [L1]13[Tb(SiW11O39)2].30H2O (SEC-1) bearing mesomorphous groups was successfully prepared by the ionic self-assembling route, exhibiting characteristic thermotropic liquid-crystalline behavior. PMID- 16041417 TI - Macrocyclic aromatic tetrasulfonamides with a stable cone conformation. AB - Aromatic tetrasulfonamide macrocycles carrying alkoxy side chains adopt a stable cone conformation in both the solid state and solution. PMID- 16041418 TI - A previously unrecognised hydronium di-cation in the crystal structure of a cucurbituril derivative. AB - Examination of the crystal structure of the adduct of hydronium tetrachloroferrate dichloride with cucurbituril resulted in the discovery that the hydronium cation present therein is not the well known H7O3+ species; rather, it is an unprecedented cyclic species of composition (H14O6)2+, templated by the formation of five hydrogen bonds to cucurbituril. PMID- 16041419 TI - High-density doxorubicin-conjugated polymeric nanoparticles via ring-opening metathesis polymerization. AB - High-density doxorubicin-conjugated polymeric nanoparticles are prepared via ring opening metathesis polymerization and sustained release of nearly 50% of the anticancer agent is observed after 24 h in mildly acidic aqueous solution. PMID- 16041420 TI - Nonlinear optical effects related to saturable and reverse saturable absorption by subphthalocyanines at 532 nm. AB - It is found that both effects of saturable absorption and reverse saturable absorption are obtained with a solution of subphthalocyanine at 532 nm depending on the intensity of 9 ns laser pulses; saturable absorption occurs at lower intensity levels whereas the reverse effect prevails at higher levels; contrary to expectations, subphthalocyanines can behave as reverse saturable absorbers at 532 nm, despite the high linear absorption at this wavelength; data have been fitted with a five-level model which considers three consecutive electronic transitions with absorption cross-section values of 1.4 x 10(-16), 1.0 x 10(-16) and 40 x 10(-16) cm(2), respectively. PMID- 16041421 TI - Synthetic gecko foot-hairs from multiwalled carbon nanotubes. AB - We report a fabrication process for constructing polymer surfaces with multiwalled carbon nanotube hairs, with strong nanometer-level adhesion forces that are 200 times higher than those observed for gecko foot-hairs. PMID- 16041422 TI - Highly enantioselective organocatalysis of the Hajos-Parrish-Eder-Sauer-Wiechert reaction by the beta-amino acid cispentacin. AB - The beta-amino acid cispentacin promotes the Hajos-Parrish-Eder-Sauer-Wiechert reaction with levels of enantioselectivity comparable to or higher than proline. PMID- 16041423 TI - Allosteric binding of anionic guests to a bicyclic host which imitates the action of a 'turnstile'. AB - A bicyclic host 1, which has a diethynyl tetrafluorophenyl axis and is expected to behave as an anion-binding 'turnstile', has been designed. PMID- 16041424 TI - The important role of solvent vapor in an organic solid state reaction. AB - Some organic reactions in the solid state proceeded very efficiently and selectively in the presence of a small amount of solvent vapor. PMID- 16041425 TI - A new synthetic route to bulky "second generation" tris(imidazol-2-ylidene)borate ligands: synthesis of a four coordinate iron(II) complex. AB - A bulky tripodal tris(carbene)borate ligand, prepared from 1-tert-butylimidazole, is cleanly transferred to iron(II) by a magnesium reagent. PMID- 16041426 TI - Formation of a long-lived charge-separated state of a zinc phthalocyanine perylenediimide dyad by complexation with magnesium ion. AB - Photoexcitation of a zinc phthalocyanine-perylenediimide (ZnPc-PDI) dyad affords the triplet excited state without the fluorescence emission, whereas addition of Mg2+ to the photoexcited ZnPc-PDI results in formation of a long-lived charge separated state (ZnPc.+-PDI.-/Mg2+) in which PDI.- forms a complex with Mg2+. PMID- 16041427 TI - Novel catalytic effects in ester aminolysis in chlorobenzene. AB - The mechanism of glyme catalyzed ester aminolysis in chlorobenzene should be modified by including a new reaction pathway that shows a first-order dependence on the concentration of the phase transfer catalyst and a second-order dependence on butylamine. PMID- 16041428 TI - Shape and size control of Ag2Se nanocrystals from a single precursor [(Ph3P)3Ag2(SeC{O}Ph)2]. AB - Monodispersed Ag2Se nanocubes and faceted nanocrystals have been synthesized by hexadecylamine (HDA) induced thermolysis of [(PPh3)3Ag2(SeC{O}Ph)2] in a mixture of TOP (tri-n-octyl phosphine) and HDA in the temperature range 95-180 degrees C. PMID- 16041429 TI - Axial bis(terpyridoxy)phosphorus(V) porphyrin: modulation of PET and EET by Zn2+ or Cd2+ ions. AB - A novel phosphorus(V) porphyrin bearing two ptp[4'-(4-phenyloxy)-2,2'ratio6',2'' terpyridine] groups was prepared and modulation of the intramolecular PET (lambda(ex)= 566 nm) and PET --> EET (lambda(ex)= 300 nm) processes was studied from ptp to phosphorus(V) porphyrin by Zn2+ or Cd2+ ions. PMID- 16041430 TI - Structure and reactivity of a new anionic N-heterocyclic carbene silver(I) complex. AB - The efficient synthesis of aN-benzoyliminoimidazolium ylide provides access to a new type of N-hetereocyclic anionic carbene, from which air stable Ag(I) and Cu(II) complexes and a catalytically active Rh(I) complex were formed. PMID- 16041431 TI - Photocatalytic overall water splitting under visible light by TaON and WO3 with an IO3-/I- shuttle redox mediator. AB - Photocatalytic water splitting into H2 and O2 under visible-light irradiation (lambda > 420 nm) is demonstrated using the oxynitride Pt-TaON for H2 evolution and a Pt-WO3 catalyst for O2 evolution in an IO3-/I- shuttle redox-mediated system. PMID- 16041432 TI - Catalytic phosphorylation using a bifunctional imidazole derived nucleophilic catalyst. AB - A bifunctional catalyst containing a polyether backbone and a nucleophilic imidazole moiety has been prepared that demonstrates cooperative catalysis in the presence of added group 1 and 2 salts for the phosphorylation of alcohols. PMID- 16041433 TI - Sterically-controlled regioselective para-substitutions of aniline. AB - Introduction of sterically demanding 1-isopropyl-2-methylpropyl or triisopropylsilyl groups at the nitrogen of aniline allows high-yielding regioselective para-substitution to be achieved using a lithiation/substitution sequence. PMID- 16041434 TI - The electrochemically-tuneable interactions between flavin-functionalised C60 derivatives and 2,6-diethylamidopyridine. AB - We report the electrochemically-tuneable interactions between flavin functionalised C60 derivatives and a diamidopyridine derivative. PMID- 16041435 TI - Single-crystalline Sb-doped SnO2 nanowires: synthesis and gas sensor application. AB - The synthesis of semiconducting Sb-doped SnO2 nanowires in mass production by an in situ doping approach are reported, and the ethanol sensing results demonstrated that Sb-doped SnO2 nanowires have a promising application for the fabrication of gas sensors with low resistance, and quick response and recovery times. PMID- 16041436 TI - A preliminary observation of additive thermodynamic contribution of pendant arms to the complexation of calixarene derivatives with mercury(II). AB - An additive thermodynamic contribution of pendant arms to the complexation of calixarene derivatives with mercury(II) in acetonitrile is for the first time demonstrated. PMID- 16041437 TI - Fabrication of polyimide nanotubes and carbon nanotubes containing magnetic iron oxide in confinement. AB - Polyimide nanotubes with tunable wall thickness were fabricated by a precursor impregnation method using an AAO template, and carbon nanotubes containing magnetic iron oxide were obtained using ferric chloride-embedded polyimide precursor by a carbonization process. PMID- 16041438 TI - A microporous scandium terephthalate, Sc2(O2CC6H4CO2)3, with high thermal stability. AB - A scandium terephthalate with isolated ScO6 octahedra and fully-linked carboxylate groups is prepared hydrothermally and possesses a novel hybrid framework structure with high thermal stability and a pore volume for N2 adsorption of 0.26 cm(3) g(-1) at 77 K. PMID- 16041439 TI - 2,2-dimethyl cyclopentanones by acid catalyzed ring expansion of isopropenylcyclobutanols. A short synthesis of (+/-)-alpha-cuparenone and (+/-) herbertene. AB - 2,2-dimethyl cyclopentanones are readily prepared by acid catalyzed ring expansion of isopropenylcyclobutanols; the method allows ready access to the family of sesquiterpenes cuparanes and herbertanes, as demonstrated by the synthesis of (+/-)-alpha-cuparenone and the direct precursor of (+/-)-herbertene. PMID- 16041440 TI - Second generation N-heterocyclic carbene-Pt(0) complexes as efficient catalysts for the hydrosilylation of alkenes. AB - A new class of benzimidazolylidene carbene-Pt(0) complexes was developed and used to efficiently catalyse the hydrosilylation of alkenes. PMID- 16041441 TI - Interpenetrated networks from a novel nanometer-sized pseudopeptidic ligand, bridging water, and transition metal ions with cds topology. AB - The combination of a new pseudopeptidic ligand, transition metal ions, and bridging water molecules results in the formation of [M(mu-TBG)(mu H2O)(H2O)2].2H2O (M: Cu, Co and H2TBG: terephthaloylbisglycine); both compounds show rare two-fold interpenetrated three-dimensional cds-nets and reversible loss of coordinated and lattice water molecules. PMID- 16041442 TI - Highly regio-, chemo- and diastereoselective synthesis of oxa-bridged spirocycles: a novel observation of reverse selectivity. AB - The reverse regio- and diastereoselectivities are observed between the reactions involving 5- and 6-membered-ring cyclic carbonyl ylide dipoles with alpha methylene ketones. A mild catalytic route to synthesize spirocyclic systems with high regio-, chemo- and diastereoselectivities is described. PMID- 16041443 TI - A new route to metallacycloalkanes. AB - Reaction of the bis-alkenyl complex cis-[Pt(PPh3)2(CH2CH2CH=CH2)2] with Grubbs 1st generation catalyst gives, in high yield, the metallacycloalkene cis [Pt(PPh3)2(CH2CH2CH=CHCH2CH2)], which can be hydrogenated to the metallacycloalkane cis-[Pt(PPh3)2(CH2)6]. PMID- 16041444 TI - Exclusive transition state stabilization in the supramolecular catalysis of Diels Alder reaction by a uranyl salophen complex. AB - Whereas the parent uranyl salophen is catalytically inactive, its phenyl derivative effectively catalyses with turnover the reaction of benzoquinone with 1,3-cyclohexadiene, while showing no appreciable affinity towards reactants and product. PMID- 16041445 TI - Non-steady-state living polymerization: a new route to control cationic ring opening polymerization (CROP) of oxetane via an activation chain end (ACE) mechanism at ambient temperature. AB - Well-defined polyoxetane with low polydispersivity has been synthesized via a novel living polymerisation process using 3-phenoxypropyl 1,4-dioxanium hexafluoroantimonate (3-PPD) as a model of a living "monomeric polyoxetane" initiator, in 1,4-dioxane at 35 degrees C. PMID- 16041446 TI - [Economic impact of chronic ischemic cardiopathy treatment in Brazil. The challenge of new cardiovascular technology inclusion]. PMID- 16041447 TI - [Annual cost of ischemic heart disease in Brazil. Public and private perspective]. AB - OBJECTIVE: To estimate the annual cost of coronary artery disease (CAD) management in Public Health Care System (SUS) and HMOs values in Brazil. METHODS: Cohort study, including ambulatory patients with proven CAD. Clinic visits, exams, procedures, hospitalizations and medications were considered to estimate direct costs. Values of appointments and exams were obtained from the SUS and the Medical Procedure List (LPM 1999) reimbursement tables. Costs of cardiovascular events were obtained from admissions in public and private hospitals with similar diagnoses-related group classifications in 2002. The price of medications used was the lowest found in the market. RESULTS: The 147 patients (65 +/- 12 years old, 63% men, 69% hypertensive, 35% diabetic and 59% with previous AMI) had an average follow-up of 24 +/- 8 months. The average estimated annual cost per patient was R$ 2,733.00, for the public sector, and R$ 6,788.00, for private and fee-for-service plans. Expenses with medications (R$ 1,154.00) represented 80% and 55% of outpatient costs, and 41% and 17% of total expenses, in public and non public sectors, respectively. The occurrence of cardiovascular event had a great impact (R$ 4,626.00 vs. R$ 1,312.00, in SUS, and R$ 13,453.00 vs. R$ 1,789.00, for HMOs, p<0.01) on the results. CONCLUSION: The average annual cost of CAD management was high, being the pharmacological treatment the main determinant of public costs. Such estimates may subsidize economical analyses in this area, and foster related healthcare policies. PMID- 16041448 TI - [Cost analysis of the treatment of acute decompensated heart failure. Levosimendan versus dobutamine]. AB - OBJECTIVE: To assess whether the treatment with levosimendan is more expensive than the usual one with dobutamine, since price of medications does not usually represent the greatest expense in the treatment of cardiac decompensation. METHODS: The cost of treatment of 18 inpatients with cardiac decompensation, 9 of which treated with dobutamine (dobuta group) and 9 with levosimendan (levo group), was compared. Groups were similar concerning age, sex, functional class and cardiac function. RESULTS: Treatment costs were similar for both groups. In the levo group, the costs with the drug were higher than in the dobuta group, but those related to the length of stay in intensive care unit and to the material used during admission were lower. Levo-drug: R$ 5,414.00; material: R$ 399.90; hospital daily rates: R$ 5,061.20; professional honorarium: R$ 3,241.80; total costs: R$ 14,117.00. Dobuta-drug: R$ 2,320.10; materials: R$ 1,665.70; hospital daily rates: R$ 6,261.90; professional honorarium: R$ 3,894.30; total costs: R$ 14,142.00. CONCLUSION: Despite the higher price of levosimendan, the global cost of the treatment was similar for patients who were treated either with dobutamine or levosimendan. Patients who were treated with levosimendan had a shorter length of stay in intensive care unit. PMID- 16041449 TI - [Postprandial lipemia: influence of aging]. AB - OBJECTIVE: To investigate the behavior of postprandial lipemia assessed by means of repeated measurements of triglyceride levels in healthy individuals aged from 20 to 50 years, divided into the following 3 age groups: GI--from 20 to 30 years; GII--from 31 to 40 years; and GIII--from 41 to 50 years. METHODS: Triglyceride levels were measured in 3 conditions: after a 12-hour fast, and 2 and 6 hours after a standard meal containing 40 g of fat. RESULTS: The repeated-measures analysis of triglyceride levels showed a distinct behavior of the age groups throughout the 6 hours. The younger participants (GI) had a reduction in the triglyceride levels in the sixth hour; the elderly (GIII) had increasing values in the sixth hour; and those in the intermediate age group (GII) maintained their triglyceride levels, when comparing the second and sixth hours of blood collection. The differences in behavior were significant (P=0.01). CONCLUSION: In a healthy adult population sample, aging influences the postprandial lipemia behavior. PMID- 16041450 TI - [Cardiovascular risk factors and mortality. Long-term follow-up (up to 20 years) in a preventive program carried out by occupational medicine]. AB - OBJECTIVE: The decrease in mortality due to cardiovascular diseases (CVD) has been achieved in the United States and the same decrease has been happening in developing countries, thanks to favorable changes in lifestyle and risk factors. Despite such consideration, the volume of information available on the distribution and behavior of that type of disease and its risk factors in Brazil is still little. METHODS: The assessment of changes in total cholesterol (TC), levels of blood pressure (BP), body mass index (BMI) and smoking was carried out, besides the occurrence of fatal (F) and non-fatal (NF) cardiovascular events, under dietary and behavioral intervention and long-term follow-up (up to 20 years) in a closed group, consisting of 621 electric power company workers of both sexes, with average age of 29.1 +/- 7.1 years old, varying from 15 to 59 years old. Actuarial curves were obtained to analyze the F and NF cardiovascular events. RESULTS: The TC mean showed significant reduction due to dietary guidance. The smoking habit decreased significantly with behavioral changes. The mean of the BPs decreased significantly with a better detection and hygienic measures, and the adhesion by definite hypertensive people to the treatment showed an index of 56.6%. On the other hand, the BMI showed an expressive and gradual increase. The probability for the individuals to remain free of any cardiovascular event was of 98.1%, whereas for the fatal events, it was 99.2%. CONCLUSION: Those results prove that initiatives directed towards the prevention must be priorities, aiming at modifying the morbimortatility rates of CVD. PMID- 16041451 TI - [Anthropometric indexes of obesity as an instrument of screening for high coronary risk in adults in the city of Salvador--Bahia]. AB - OBJECTIVE: To compare some anthropometric indexes of obesity and identify among them which one best discriminates the high coronary risk (HCR). METHODS: A cross section study, with sample consisting of 968 adults, between 30 and 74 years old, being 391 (40.4%) men. Many Receiver Operating Characteristic (ROC) curves were obtained and compared to areas under them among the conicity index (C index), body mass index (BMI), waist-hip circumference ratio (WHCR), waist circumference (WC) and HCR. The sensitivity and specificity to identify and compare the best cut-off point among the many indexes of obesity to discriminate the HCR were also identified. A confidence interval of 95% was used. RESULTS: The largest area under ROC curve was found between the C index and the HCR, in individuals of male sex, 0.80 (0.74-0.85), significantly differing from the other indexes of obesity. In women, the largest area found under the ROC curve was 0.76 (0.71-0.81), being equal between C, WHCR and HCR indexes. CONCLUSION: Those results show that C and WHCR indexes are the best indexes of obesity to discriminate HCR. WC has intermediate discriminatory power and the BMI was the least suitable anthropometric index of obesity to discriminate HCR. Those data suggest that the indexes of abdominal obesity are better to discriminate HCR than the indexes of general obesity. PMID- 16041452 TI - [Weaning from mechanical ventilation by using pressure support or T-tube ventilation. Comparison between patients with and without heart disease]. AB - OBJECTIVE: To assess cardiorespiratory variables during weaning from mechanical ventilation by using the T-tube and pressure support techniques, and to compare them in groups of patients with and without heart disease. METHODS: To assess the following parameters of 20 patients (57 +/- 15 years) undergoing weaning from mechanical ventilation: oxygenation; CO2 elimination; respiratory and heart rates; tidal and minute volumes; blood pressure; and electrocardiographic alterations. Data were recorded by using both techniques at the following times: zero, 15, and 30 minutes, and after a 30-minute interval. The patients were divided into 2 groups, with heart disease (n=11) and without heart disease (n=9), and then compared. RESULTS: The pressure support ventilation showed significantly more elevated oxygenation and CO2 elimination values, and reduced respiratory rate as compared with those of the T-tube ventilation. No difference was found in regard to blood pressure and heart rate. More patients with heart disease had alterations in the ST segment [7 (64%) patients versus 2 (22%)] and arrhythmias [3 (27%) versus 1 (11%)], as compared with patients without heart disease. A lower frequency of tachycardia was observed in those with heart disease. CONCLUSION: When comparing pressure support ventilation with T-tube ventilation, a better response was observed in the measurements of the respiratory and oxygenation parameters when using pressure support ventilation. No significant difference was observed in the measurements of cardiovascular parameters. In both weaning techniques, patients with heart disease had tachycardia less frequently, more alterations in the ST segment, and a greater tendency towards the occurrence of arrhythmias. PMID- 16041453 TI - [Surgical strategy for transposition of the great arteries with intact ventricular septum after the neonatal period]. AB - OBJECTIVE: To assess the surgical results in patients with transposition of the great arteries and intact ventricular septum undergoing surgery after the neonatal period. METHODS: From January 1998 to March 2004, 121 children with transposition of the great arteries with intact ventricular septum were treated, 29 (24%) of whom after the neonatal period. Selection for surgical treatment was based on echocardiographic assessment by use of the calculation of left ventricular mass and configuration of the ventricular septum. Of the 29 children, 12 were selected for primary anatomic correction, 12 for 2-stage correction after surgical preparation of the left ventricle, and 5 underwent atrial correction. RESULTS: In the group undergoing primary anatomic correction, one (8.3%) patient died at the hospital due to sepsis. In the group undergoing 2-stage correction, 5 patients underwent slow preparation with correction 3-6 months after the first stage, 4 of whom died after the first stage. This fact caused a change in our protocol, with adoption of the rapid preparation technique in the other 7 patients, of whom all achieved the second stage. Of the 8 children undergoing the second stage, one died at the hospital and another died later. Late clinical evolution of the children in both groups is excellent. CONCLUSION: The echocardiographic selection allowed a safe choice of the best surgical approach for patients with transposition of the great arteries and intact ventricular septum after the neonatal period. PMID- 16041454 TI - [Maximal heart rate in exercise tests on treadmill and in a cycloergometer of lower limbs]. AB - OBJECTIVE: To compare, retrospectively, the values of maximum heart rate (MHR) and the decrease of the heart rate at the first minute of recovery, which were obtained in an exercise test (ET) performed in two different ergometers and at different moments. METHODS: Sixty individuals (from 29 to 80 years old), submitted to cardiopulmonary ET in a cycle of lower limbs (CLL) in our laboratory and who had previous ET (up to 36 months) in a treadmill (TRM) in other laboratories, under identical conditions of medications of negative chronotropic action. RESULTS: MHR was similar in CLL: 156 +/- 3 and TRM: 154 +/- 2 bpm (p=0.125), whereas dHR was higher in CLL: 33 +/- 2, EST: 26 +/- 3 bpm (mean +/- standard error of the mean) (p<0.001). In hemodynamic variables studied, the systolic blood pressure and the double product were higher in the ET-CLL (p<0.001). The electrocardiogram (ECG) was similar in both ETs, except due to more frequent supraventricular arrhythmias in CLL. CONCLUSION: a) With some diligence from the examiner and previous knowledge of MHR in a previous ET it is possible to obtain high levels of MHR in an ET-CLL; b) interrupting the MHR-based ET forecast through equations tends to lead to sub-maximum efforts; c) dHR differs in active and passive recoveries; d) new ways to analyze the HR behavior under exercise, which is not only the value of MHR, are necessary to characterize an ET as maximum. PMID- 16041455 TI - [Coarctation of the aorta in infants under one year of age. An analysis of 20 years of experience]. AB - OBJECTIVE: A review of experience with techniques of correction used, in the last 20 years, in children younger than one year old. METHODS: In the period from 1978 to 1998, 148 patients (pt) with coarctation of the aorta (CoAo), under one year of age, with or without associated intracardiac defects, were submitted to surgery. Median age 50 days, 92 female pt (62.1%). The average weight was 4,367 +/- 1,897 gr. The average follow-up was 1,152 +/- 1,462 days. The population was divided in 3 groups: Group I, isolated CoAo: 74 pt (50%); Group II, CoAo and interventricular communication (IVC): 41 pt (27.7%) and Group III, CoAo with complex intracardiac malformations: 33 pt (22.3%). RESULTS: The total mortality was of 43 patients (29%). In patients younger than 30 days, the mortality was 53%, p=0.009, DR=4.5, between 31 and 90 days, 14.7%, p=0.69, and over 91 days, 15%, p=0.004. The probability of actuarial survival of the whole population was 67% at 5 and 10 years. Thirty-six patients (24.3%) had recoarctation, from which 18 patients (50%) were younger than 30 days, DR=6.35. The incidence of recoarctation was with Waldhausen technique in 4 patients (10%) and with the classic termino-terminal technique in 19 patients (26%) p=0.03, and isthmusplastic operation in 6 patients (37.5%). The patients younger than 30 days showed a relative risk for recoarctation de DR=6.35. The probability of actuarial survival, free of coarctation repair, at 5 and 10 years was of 69% with Waldhausen's technique and 63% with the classic termino-terminal technique. CONCLUSION: Patients younger than 30 days showed increased mortality and recoarctation risk. Waldhausen's technique in patients older than 30 days showed effective. The classic termino-terminal technique did not show to be a good option in all age ranges, being imperative to carry out more radical technical variations, such as the extended termino-terminal. PMID- 16041456 TI - [Lipid profile and risk factors for cardiovascular diseases in medicine students]. AB - OBJECTIVE: To analyze the lipid profile and its correlation with risk factors for cardiovascular diseases (CVD) among medicine students. METHODS: We assessed 153 students, regardless of sex, with age between 18 and 31 years old, submitted to lipid profile analysis, including serum levels of total cholesterol (TC), fraction of cholesterol of low (LDLc), high (HDLc) and very low density (VLDLc) lipoproteins and triglycerides (TG), besides of life habits and anthropometric data. Statistic analysis was employed, including the test of Mann Whitney, chi square test, Pearson's correlation and multivariate analysis, by adopting a significance level for a value of p<0.05. RESULTS: Sedentary lifestyle (43.1%) and familial history for CVD, particularly hypertension (74.5%), were distinguished. The lipid profile showed desirable, although altered levels of TC, LDLc and TG were detected in 11.8%, 9.8% and 8.5% from the students, respectively, and reduced levels of HDLc in 12.4% of them. Women showed significantly reduced values for LDLc and high values for HDLc compared to men's (p=0.031 and p<0.0001, respectively). There was a significant association between lipid profile and, preferably, body mass index (BMI), sedentary lifestyle, ingestion of alcohol, contraceptive use, familial antecedents of cerebrovascular accident and dyslipidemia. CONCLUSION: Familial history for CVD, sedentary lifestyle and contraceptive use among medicine students showed frequent and associated to the lipid profile, as well as ingestion of alcohol and BMI. Although with desirable lipid profile, regardless of sex, higher levels of LDLc and reduced levels of HDLc in male sex provide a disadvantage to men compared to women. PMID- 16041457 TI - [Eikenella corrodens infective endocarditis]. AB - The HACEK microorganisms (Haemophilus spp, Actinobacillus actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens, and Kingella kingae) account for 3% of the cases of endocarditis. They have the following similar clinical and microbiological properties: are Gram-negative bacilli, more easily isolated in aerobic media; their cultures require prolonged incubation time for growing (mean, 3.3 days); and may be considered part of normal flora of upper respiratory tract and oropharynx. The following characteristics have been identified in endocarditis caused by the HACEK microorganisms: insidious clinical findings; difficult diagnosis due to the fastidious nature of the microorganisms; and negative cultures. The Eikenella corrodens endocarditis was first described in 1972. That microorganism continues to be a rare etiological agent. We report the case of a female patient with native valve, who had Eikenella corrodens infective endocarditis. PMID- 16041458 TI - [Right atrial aneurysm associated to fetal hydrops: diagnosis through fetal echocardiography]. AB - Right atrium aneurysms are entities which are rarely reported in cardiologic practice, especially in intrauterine life, and may be mistaken with pericardial effusion and Ebstein's anomaly. We show a review of the literature and illustrate with a case of prenatal diagnosis of right atrium aneurysm running through with hydropsy signs. PMID- 16041459 TI - [Prevailing right ventricular myocardiopathy for previous myocarditis or arrhythmogenic dysplasia?]. AB - A clinical case of a 10-year-old male patient is reported. His dilated and prevailing right ventricular myocardiopathy shows diagnostic difficulties between previous myocarditis etiology and arrhythmogenic dysplasia. As the elements are not pathognomonic of one or other cause, the increase of cardiac enzymes in subacute stage maybe tends to the supposition of previous myocarditis. Hence, the questioning that many cases labeled as arrhythmogenic dysplasia can truly correspond to the possibility of evolutional myocarditis. The controversial clinic management is disputable. PMID- 16041460 TI - [Therapeutics in clinical cardiovascular practice. Experiences and evidence]. PMID- 16041461 TI - [Thrombolytic therapy in prosthetic valve thrombosis]. PMID- 16041462 TI - A practical guide to the management of menopausal symptoms in breast cancer patients. AB - Breast cancer is the most frequently diagnosed cancer in Canadian women. As a result of increased screening and improved treatment, more women are becoming long-term breast cancer survivors. However, due to either their treatment or prolonged survival, many of these women now have to face the consequences of premature menopause and prolonged estrogen deprivation. Hormone replacement therapy/estrogen replacement therapy (HRT/ERT) has, in the past, been recommended to healthy women at menopause not only for relief of short-term menopausal changes, particularly hot flashes, but also for its benefits on bone density, fracture reduction, and genitourinary symptoms. Recent studies have demonstrated that not only is HRT associated with an increased risk of developing breast cancer, but it also has been shown to increase the risk of recurrence in those with a breast cancer history. Until the safety of HRT/ERT in breast cancer patients can be more fully clarified, it would be wise to develop alternative strategies for the management of menopausal symptoms in these patients. This paper will discuss nonestrogen-based therapies for hot flashes, osteoporosis, and genitourinary symptoms, with emphasis on efficacy and safety in breast cancer survivors. PMID- 16041463 TI - Systematic review of specialist palliative day-care for adults with cancer. AB - GOAL OF WORK: To inform future practice, research and policy in specialist palliative day-care by systematically reviewing the evidence for how the structure and process of this form of care relate to outcomes for adults with cancer. PATIENTS AND METHODS: Medical, nursing and social science computerized databases were searched up until December 2003 for studies of palliative day-care reporting information on service structure, care processes or outcomes including symptom control, quality of life, social and psychological support, and patient and relative satisfaction with care. Qualitative and quantitative studies were assessed for methodological quality and graded, and the findings synthesized into the review. MAIN RESULTS: Twelve observational studies were found that showed the difficulties of evaluating a service already operating and of recruiting a vulnerable population of patients as they deteriorated. Most services are nurse led, but varied in the facilities, staff mix, care models, activities and places they offered. Patients attending seemed a selected group of those already receiving palliative care who were mostly white, aged over 60 years and retired, with needs for emotional and social support and pain control. There were insufficient studies to provide conclusive evidence of improved symptom control or health-related quality of life, but all qualitative studies found patients valued the social support and opportunity to take part in activities that day care provided. CONCLUSIONS: There is evidence for high satisfaction among patients selected into day-care, but not yet sufficient to judge whether this improves symptom control or health-related quality of life. Further research should explore selection and access into care, the most effective models of care, its cost, and potential benefits for relatives and carers. Comparison with models of care for older adults and those with mental illness would also be informative. PMID- 16041464 TI - Support group for cancer patients. Does it improve their physical and psychological wellbeing? A pilot study. AB - In order to evaluate a structured support intervention programme, symptoms and quality of life (QOL) were studied in 61 consecutive cancer patients with different tumour diagnoses in relation to intervention and follow-up. The majority of the patients were female. The Edmonton symptom assessment scale (ESAS), the Norwegian fatigue questionnaire and the hospital anxiety and depression scale (HADS) were used. Data were analysed according to ANOVA and Tukey's honest significant difference (HSD) test. ESAS sickness score (P=0.0001), depression (P=0.0001), anxiety (P=0.0001) and QOL (P=0.0009) improved and the improvements in depression and anxiety were still significant after 3 months (P=0.02 respectively). Aspects of fatigue also improved significantly in 7 of 11 questions after the intervention (P< or =0.04) but these improvements were not observed after 3 months. Anxiety according to the HADS questionnaire improved significantly after the intervention (P=0.0006). The majority of the patients highly appreciated the possibility of sharing thoughts and troubles with others with similar experiences (59%). The programme was therefore found to improve QOL, and physical and psychological functions. Randomized studies in relation to immunological changes and follow-up are in progress. PMID- 16041465 TI - Intraventricular haemorrhage from a renal cell carcinoma pituitary metastasis. AB - Symptomatic pituitary metastasis and intraventricular haemorrhage from a cerebral metastasis are exceptional events in the natural history of a renal cell carcinoma. We report the first case of a metastatic renal cell carcinoma to the pituitary gland presenting with intraventricular haemorrhage. The origin of intraventricular haemorrhage and its association with renal cell carcinoma pituitary metastasis are discussed. PMID- 16041466 TI - Haemorrhagic central neurocytoma of the fourth ventricle. PMID- 16041467 TI - The risk of rebleeding after external lumbar drainage in patients with untreated ruptured cerebral aneurysms. AB - BACKGROUND: Does continuous external lumbar CSF drainage before aneurysm repair in patients with aneurysmal subarachnoid hemorrhage increase the risk of rebleeding? METHOD: The study population, consisting of 18 patients treated by External Lumbar Drainage (ELD) after SAH before aneurysm repair, was compared with an independent control group of 324 SAH patients treated in another clinic. Control patients were selected for not being treated for the ruptured aneurysm yet and not having undergone any form of CSF drainage during the exposure time in the case patients. We calculated hazard ratios with the Cox regression model, adjusted for age and clinical condition on admission and hydrocephalus. FINDINGS: The cox regression analysis shows a non-adjusted hazard ratio of 2.1 (95% CI 0.8 to 5.3) in the model with 5 rebleedings in 18 patients. Adjustment for age, clinical condition on admission and hydrocephalus did not alter the hazard ratio estimate importantly in either analyses. CONCLUSIONS: An increased risk of rebleeding by external lumbar drainage in the acute phase after aneurysmal SAH could not be confirmed, but the data are too imprecise to rule out an increased risk. The potential benefits of early drainage should be weighed against the risks if the aneurysm is not occluded before or early after the start of drainage. PMID- 16041468 TI - Resolution of superior oblique myokymia following microvascular decompression of trochlear nerve. PMID- 16041469 TI - The predictive value of external continuous lumbar drainage, with cerebrospinal fluid outflow controlled by medium pressure valve, in normal pressure hydrocephalus. AB - BACKGROUND: Although sporadic studies have described temporary external cerebrospinal fluid (CSF) lumbar drainage as a highly accurate test for predicting the outcome after ventricular shunting in normal pressure hydrocephalus (NPH) patients, a more recent study reports that the positive predictive value of external lumbar drainage (ELD) is high but the negative predictive value is deceptively low. Therefore, we conducted a prospective study in order to evaluate the predictive value of a continuous ELD, with CSF outflow controlled by medium pressure valve, in NPH patients. METHOD: Twenty-seven patients with presumed NPH were admitted to our department and CSF drainage was carried out by a temporary (ELD) with CSF outflow controlled by a medium pressure valve for five days. All patients received a ventriculoperitoneal shunt using a medium pressure valve based upon preoperative clinical and radiographic criteria of NPH, regardless of ELD outcome. Clinical evaluation of gait disturbances, urinary incontinence and mental status, and radiological evaluation with brain CT was performed prior to and after ELD test, as well as three months after shunting. FINDINGS: Twenty-two patients were finally shunted and included in this study. In a three-month follow-up, using a previously validated score system, overall improvement after permanent shunting correlated well to improvement after ELD test (Spearman's rho = 0.462, p = 0.03). When considering any degree of improvement as a positive response, ELD test yielded high positive predictive values for all individual parameters (gait disturbances 94%, 95% CI 71%-100%, urinary incontinence 100%, 95% CI 66%-100%, and mental status 100%, 95% CI 66% 100%) but negative predictive values were low (< 50%) except for cognitive impairment (85%, 95% CI 55%-98%). CONCLUSION: This study suggests that a positive ELD-valve system test should be considered a reliable criterion for preoperative selection of shunt-responsive NPH patients. In case of a negative ELD-valve system test, further investigation of the presumed NPH patients with additional tests should be performed. PMID- 16041470 TI - Hardware-related complications of deep brain stimulation: a ten year experience. AB - OBJECTIVE: To analyse the occurrence of hardware-related complications in patients with deep brain stimulation (DBS), over a long period of time. METHOD: All patients operated on with DBS at our institution between 1993 and 2002 were followed with respect to adverse events related to the implanted hardware. RESULTS: One hundred and nineteen consecutive patients underwent 139 procedures with implantation of 161 electrodes. The minimum follow-up was 12 months. The follow-up time was 540 electrode-years. The rate of hardware-related complications per electrode-year was 4.3%. In total, 17 patients (15%) had 23 hardware-related complications. These included 8 electrode breakages, 4 electrode migrations, 2 stimulator migrations, 3 erosions, 2 erosions and infections, 2 infections and 2 cases of stimulator malfunction. The majority of these complications occurred during the first four years in our experience. CONCLUSIONS: DBS is a life-long therapy that requires a life-long follow-up. Increased experience and adaptation of surgical technique are the main determinants for avoidance of hardware-related complications. PMID- 16041471 TI - Infarction of the right hemisphere in a patient with antiphospholipid antibody syndrome. AB - Hemicraniectomy as a surgical treatment for intracranial pressure following large ischemic lesions is widely practiced in selected patients. The antiphospholipid antibody syndrome (APS), a disorder characterized by recurrent arterial and venous thrombosis, is a very rare cause of space occupying ischemic lesions. We present a case of a 35 year old female diagnosed with APS who initially presented with small ischemic lesions and within days developed a massive near-total infarction of the right hemisphere. Because of central nervous system, skin and systemic manifestations Sneddon's syndrome and catastrophic antiphospholipid antibody syndrome (CAPS) remained a possible diagnoses. Sneddon's syndrome is a non-inflammatory occlusive arteriopathy of small and medium size arteries predominantly of the skin and brain, whereas the catastrophic antiphospholipid antibody syndrome is characterized by acute multi-organ system thrombosis of small and large vessels. In addition to the diagnostic criteria for APS a heterozygous factor V Leiden mutation was found in this patient, which may be a contributing risk factor for cerebral ischemia. When considering invasive decompressive procedures the neurosurgeon has to be aware of the poor prognosis of some forms of APS with systemic manifestations. PMID- 16041472 TI - Now and later? The sticky question of lymph node management in patients receiving preoperative chemotherapy. PMID- 16041473 TI - Sentinel lymph node biopsy for cutaneous melanoma of the head and neck. AB - BACKGROUND: Lymph node status is the most important prognostic factor for patients with cutaneous melanoma. Sentinel lymph node biopsy (SLNB) is now the standard of care for staging clinically node-negative patients. It is accurate with low morbidity, yet SLNB for head and neck melanoma is challenging because of unpredictable lymphatic drainage and risk of complications. METHODS: A retrospective analysis of prospectively collected data identified patients with cutaneous melanoma of the head and neck > or =.76 mm. Sentinel lymph nodes were identified by using a standardized protocol of preoperative lymphoscintigrams, intraoperative blue dye injections, and handheld gamma probes. Clinical, surgical, and pathologic data were collected and analyzed. RESULTS: A sentinel lymph node was removed in 41 (94%) of 44 patients. Seven (17%) of 41 had at least 1 positive sentinel lymph node. Three of seven had primary tumors <1 mm (two of the three were not ulcerated). The sites of lymphatic drainage of the primary lesion were discordant, with historical anatomically predicted sites in 24.4% of cases. None of the 34 patients with negative SLNB has had a nodal recurrence (false-negative rate, 0%; sensitivity and negative predictive value, 100%). The mean follow-up is 22.4 months (range, <1-69 months). Seven (17%) of 41 patients had minor complications. CONCLUSIONS: SLNB in the head and neck area is challenging; however, combined preoperative, intraoperative, and histological techniques produce a sensitive procedure with a high negative predictive value. The lack of false-negative results obviates the need for prophylactic neck dissections. PMID- 16041474 TI - Ontogenetic development of the gastrointestinal microbiota in the marine herbivorous fish Kyphosus sydneyanus. AB - Molecular techniques were used to investigate the composition and ontogenetic development of the intestinal bacterial community in the marine herbivorous fish Kyphosus sydneyanus from the north eastern coast of New Zealand. Previous work showed that K. sydneyanus maintains an exclusively algivorous diet throughout post-settlement life and passes through an ontogenetic diet shift from a juvenile diet which is readily digestible to an adult diet high in refractory algal metabolites. Terminal restriction fragment length polymorphism (T-RFLP) analysis was used to investigate the relationship between bacterial community structure and fish size. Bacterial diversity was higher in posterior gut sections than anterior gut sections, and in larger fish than in smaller fish. Partial sequencing of bacterial 16S rDNA genes PCR amplified and cloned from intestine content samples was used to identify the phylogenetic affiliation of dominant gastrointestinal bacteria. Phylogenetic analysis of clones showed that most formed a clade within the genus Clostridium, with one clone associated with the parasitic mycoplasmas. No bacteria were specific to a particular intestinal section or size class of host, though some appeared more dominant than others and were established in smaller fishes. Clones closely related to C. lituseburense were particularly dominant in most intestine content samples. All bacteria identified in the intestinal samples were phylogenetically related to those possessing fermentative type metabolism. Short-chain fatty acids in intestinal fluid samples increased from 15.6 +/- 2.1 mM in fish <100 mm to 51.6 +/- 5.5 mM in fish >300 mm. The findings of this study support the hypothesis that the ontogenetic diet shift of K. sydneyanus is accompanied by an increase in the diversity of intestinal microbial symbionts capable of degrading refractory algal metabolites into short-chain fatty acids, which can then be assimilated by the host. PMID- 16041475 TI - Improved fatty acid and leukotriene pattern with a novel lipid emulsion in surgical patients. AB - OBJECTIVE: We assessed the effects of a novel lipid emulsion with reduced content of n-6 fatty acids (FA), increased share of MUFA and n-3 FA and supplemental vitamin E on fatty acid and leukotriene pattern in surgical patients. METHODS: In a double-blind, randomized study 33 patients received isonitrogenous, isocaloric TPN over 5 postoperative days following major abdominal surgery. 19 patients received the new SMOFlipid 20% and 14 patients a standard soybean oil emulsion (Lipovenoes 20%, both Fresenius Kabi), each 1.5 g fat/kg body weight (BW)/d. Routine lipid biochemistry, plasma tocopherol, fatty acid pattern in plasma phospholipids, as well as leukotriene (LT) release in leukocytes were assessed. Additionally, fatty acid pattern in leukocyte and platelet phospholipids were analysed, but results are not presented. RESULTS: On day 6, plasma alpha tocopherol (34.2 +/- 10.3 vs. 17.6 +/- 2.9 micromol/L) and, in plasma PL, total n 3 FA were higher (11.1 +/- 1.9 vs. 4.9 +/- 0.9 mol%; p < 0.05) and total n-6 FA lower (23.8 +/- 2.2 vs. 31.8 +/- 1.7 mol%; P < 0.05); the ratio n-3/n-6 FA being elevated (0.5 +/- 0.1 vs. 0.2 +/- 0.0 p < 0.05) with SMOFlipid compared to the soybean oil emulsion. The shares of EPA (3.3+/-1.0 vs. 0.4+/-0.2 mol%; p<0.05) and DHA (6.9 +/- 1.8 vs. 3.7 +/- 0.8 mol%; p < 0.05) were highly increased but that of arachidonic acid (AA) was unchanged with SMOFlipid while the ratio EPA/AA was increased (0.7 +/- 0.2 vs. 0.1 +/- 0.0 p < 0.05). LTB(5) release was enhanced on day 6 (8.1 +/- 5.3 vs. 1.8 +/- 3.8 pmol/10(7) PMN, p < 0.05) and liberation of LTB(4) was lowered, yet not significantly with SMOFlipid (124.0 +/- 51.2 vs. 152.1 +/- 68.8 pmol/10(7) PMN). Length of hospital stay was significantly shorter with SMOFlipid (13.4 +/- 2.0 vs. 20.4 +/- 10.0 days, p < 0.05). CONCLUSION: Treatment with the new emulsion SMOFlipid is well tolerated and modulates FA and leukotriene pattern suggesting favourable anti-inflammatory effects and further clinical benefits. PMID- 16041476 TI - Isolation of uncultivated anaerobic thermophiles from compost by supplementing cell extract of Geobacillus toebii in enrichment culture medium. AB - Several researchers have reported that microorganisms can be cultivated only in the presence of other microorganisms. We suggest that a portion of uncultivated microorganisms might be cultivated in the presence of cellular components released from bacteria in their natural environments. In this study, the cell extract of Geobacillus toebii was used to enrich uncultivated thermophiles from compost. In the process of enrichment cultures, cell extract supplementation apparently changed the community composition. This change was monitored by PCR DGGE targeting 16S rRNA gene. Five novel groups of microorganisms (similarity of 16S rRNA gene to the closest relative <96%) were specifically isolated from enrichment cultures by using cell extract-supplemented culture media. Their growth was found to be dependent on the addition of extract of G. toebii. Putting these findings together, we suggest that the extracts of bacteria could be one of the growth factors in the thermal ecosystem with a possibility of extending other ecological niches. PMID- 16041477 TI - Oxidation of thiosulfate to tetrathionate by an haloarchaeon isolated from hypersaline habitat. AB - A novel, extremely halophilic, neutrophilic archaeon was isolated from a mixed sediment sample from different hypersaline lakes in Kulunda steppe (Altai, Russia) at 4 M NaCl with acetate and thiosulfate as substrates. The enrichment culture developed in two phases. During the first phase, a rapid growth of heterotrophic, red-colored, polymorphic rods occurred with the concomitant oxidation of thiosulfate to tetrathionate. The latter was subsequently oxidized to sulfate during a second, slower phase by extremely halophilic, chemolithoautotrophic bacteria belonging to the gamma subdivision of the Proteobacteria. The archaeal strain HG 1 was isolated from the first phase of the enrichment culture using acetate as substrate. It was able to oxidize thiosulfate to tetrathionate during heterotrophic growth with acetate-a property not yet demonstrated for any of the known haloarchaea. The presence of tetrathionate synthase, the enzyme responsible for thiosulfate oxidation, was detected in strain HG 1. The activity was associated with membranes and depended specifically on Cl-, in contrast to the similar activity in extremely halophilic sulfur oxidizing Gammaproteobacteria from the same enrichment, which was soluble and demanded both Na+ and Cl- . Strain HG 1 was identified as a member of the genus Natronorubrum. PMID- 16041478 TI - BioMagResBank databases DOCR and FRED containing converted and filtered sets of experimental NMR restraints and coordinates from over 500 protein PDB structures. AB - We present two new databases of NMR-derived distance and dihedral angle restraints: the Database Of Converted Restraints (DOCR) and the Filtered Restraints Database (FRED). These databases currently correspond to 545 proteins with NMR structures deposited in the Protein Databank (PDB). The criteria for inclusion were that these should be unique, monomeric proteins with author provided experimental NMR data and coordinates available from the PDB capable of being parsed and prepared in a consistent manner. The Wattos program was used to parse the files, and the CcpNmr FormatConverter program was used to prepare them semi-automatically. New modules, including a new implementation of Aqua in the BioMagResBank (BMRB) software Wattos were used to analyze the sets of distance restraints (DRs) for inconsistencies, redundancies, NOE completeness, classification and violations with respect to the original coordinates. Restraints that could not be associated with a known nomenclature were flagged. The coordinates of hydrogen atoms were recalculated from the positions of heavy atoms to allow for a full restraint analysis. The DOCR database contains restraint and coordinate data that is made consistent with each other and with IUPAC conventions. The FRED database is based on the DOCR data but is filtered for use by test calculation protocols and longitudinal analyses and validations. These two databases are available from websites of the BMRB and the Macromolecular Structure Database (MSD) in various formats: NMR-STAR, CCPN XML, and in formats suitable for direct use in the software packages CNS and CYANA. PMID- 16041479 TI - Linear analysis of carbon-13 chemical shift differences and its application to the detection and correction of errors in referencing and spin system identifications. AB - Statistical analysis reveals that the set of differences between the secondary shifts of the alpha- and beta-carbons for residues i of a protein (Deltadelta13C(alpha)i - Deltadelta13C(beta)i) provides the means to detect and correct referencing errors for 1H and 13C nuclei within a given dataset. In a correctly referenced protein dataset, linear regression plots of Deltadelta13C(alpha)i, Deltadelta13C(beta)i, or Deltadelta1H(alpha)i vs. (Deltadelta13C(alpha)i - Deltadelta13C(beta)i) pass through the origin from two directions, the helix-to-coil and strand-to-coil directions. Thus, linear analysis of chemical shifts (LACS) can be used to detect referencing errors and to recalibrate the 1H and 13C chemical shift scales if needed. The analysis requires only that the signals be identified with distinct residue types (intra residue spin systems). LACS allows errors in calibration to be detected and corrected in advance of sequence-specific assignments and secondary structure determinations. Signals that do not fit the linear model (outliers) deserve scrutiny since they could represent errors in identifying signals with a particular residue, or interesting features such as a cis-peptide bond. LACS provides the basis for the automated detection of such features and for testing reassignment hypotheses. Early detection and correction of errors in referencing and spin system identifications can improve the speed and accuracy of chemical shift assignments and secondary structure determinations. We have used LACS to create a database of offset-corrected chemical shifts corresponding to nearly 1800 BMRB entries: 300 with and 1500 without corresponding three-dimensional (3D) structures. This database can serve as a resource for future analysis of the effects of amino acid sequence and protein secondary and tertiary structure on NMR chemical shifts. PMID- 16041480 TI - Sensitivity enhancement in NMR of macromolecules by application of optimal control theory. AB - NMR of macromolecules is limited by large transverse relaxation rates. In practice, this results in low efficiency of coherence transfer steps in multidimensional NMR experiments, leading to poor sensitivity and long acquisition times. The efficiency of coherence transfer can be maximized by design of relaxation optimized pulse sequences using tools from optimal control theory. In this paper, we demonstrate that this approach can be adopted for studies of large biological systems, such as the 800 kDa chaperone GroEL. For this system, the 1H-15N coherence transfer module presented here yields an average sensitivity enhancement of 20-25% for cross-correlated relaxation induced polarization transfer (CRIPT) experiments. PMID- 16041481 TI - Bacterial expression and one-step purification of an isotope-labeled heterotrimeric G-protein alpha-subunit. AB - Heterologous expression systems are often employed to generate sufficient quantities of isotope-labeled proteins for high-resolution NMR studies. Recently, the interaction between the prodomain region of subtilisin and an active, mutant form of the mature enzyme has been exploited to develop a cleavable affinity tag fusion system for one-step generation and purification of full-length soluble proteins obtained by inducible prokaryotic expression. As a first step towards applying high-resolution NMR methods to study heterotrimeric G-protein alpha subunit (G(alpha)) conformation and dynamics, the utility of this subtilisin prodomain fusion system for expressing and purifying an isotope-labeled G(alpha) chimera (approximately 40 kDa polypeptide) has been tested. The results show that a prodomain fused G(alpha) chimera can be expressed to levels approaching 6-8 mg/l in minimal media and that the processed, mature protein exhibits properties similar to those of G(alpha) isolated from natural sources. To assay for the functional integrity of the purified G(alpha) chimera at NMR concentrations and probe for changes in the structure and dynamics of G(alpha) that result from activation, 15N-HSQC spectra of the GDP/Mg2+ bound form of G(alpha) obtained in the absence and presence of aluminum fluoride, a well known activator of the GDP bound state, have been acquired. Comparisons of the 15N-HSQC spectra reveals a number of changes in chemical shifts of the 1HN, 15N crosspeaks that are discussed with respect to expected changes in the protein conformation associated with G(alpha) activation. PMID- 16041482 TI - Error estimation and global fitting in transverse-relaxation dispersion experiments to determine chemical-exchange parameters. AB - Off-resonance effects can introduce significant systematic errors in R2 measurements in constant-time Carr-Purcell-Meiboom-Gill (CPMG) transverse relaxation dispersion experiments. For an off-resonance chemical shift of 500 Hz, 15N relaxation dispersion profiles obtained from experiment and computer simulation indicated a systematic error of ca. 3%. This error is three- to five fold larger than the random error in R2 caused by noise. Good estimates of total R2 uncertainty are critical in order to obtain accurate estimates in optimized chemical exchange parameters and their uncertainties derived from chi2 minimization of a target function. Here, we present a simple empirical approach that provides a good estimate of the total error (systematic + random) in 15N R2 values measured for the HIV protease. The advantage of this empirical error estimate is that it is applicable even when some of the factors that contribute to the off-resonance error are not known. These errors are incorporated into a chi2 minimization protocol, in which the Carver-Richards equation is used fit the observed R2 dispersion profiles, that yields optimized chemical exchange parameters and their confidence limits. Optimized parameters are also derived, using the same protein sample and data-fitting protocol, from 1H R2 measurements in which systematic errors are negligible. Although 1H and 15N relaxation profiles of individual residues were well fit, the optimized exchange parameters had large uncertainties (confidence limits). In contrast, when a single pair of exchange parameters (the exchange lifetime, tau(ex), and the fractional population, p(a)), were constrained to globally fit all R2 profiles for residues in the dimer interface of the protein, confidence limits were less than 8% for all optimized exchange parameters. In addition, F-tests showed that quality of the fits obtained using tau(ex), p(a) as global parameters were not improved when these parameters were free to fit the R2 profiles of individual residues. Finally, nearly the same optimized global tau(ex), p(a) values were obtained, when the 1H and 15N data sets for residues in the dimer interface, were fit independently; the difference in optimized global parameters, ca. 10%, was of marginal significance according to the F-test. PMID- 16041483 TI - High-resolution aliphatic side-chain assignments in 3D HCcoNH experiments with joint H-C evolution and non-uniform sampling. AB - We describe an efficient NMR triple resonance approach that correlates, at high resolution, protein side-chain and backbone resonances. It relies on the combination of two strategies: joint evolution of aliphatic side-chain proton/carbon coherences using a backbone N-H based HCcoNH reduced dimensionality (RD) experiment and non-uniform sampling (NUS) in two indirect dimensions. A typical data set containing such correlation information can be acquired in 2 days, at very high resolution unfeasible for conventional 4D HCcoNH-TOCSY experiments. The resonances of the aliphatic side-chain protons are unambiguously assigned to their attached carbons through the analysis of the 'sum' and 'difference' spectra. This approach circumvents the tedious process of manual resonance assignments using HCcH-TOCSY data, while providing additional resolving power of backbone N-H signals. A simple peak-list based algorithm has been implemented in the IBIS software for rapid automated backbone and side-chain assignments. PMID- 16041484 TI - Managing the solvent water polarization to obtain improved NMR spectra of large molecular structures. AB - In large molecular structures, the magnetization of all hydrogen atoms in the solute is strongly coupled to the water magnetization through chemical exchange between solvent water and labile protons of macromolecular components, and through dipole-dipole interactions and the associated "spin diffusion" due to slow molecular tumbling. In NMR experiments with such systems, the extent of the water polarization is thus of utmost importance. This paper presents a formalism that describes the propagation of the water polarization during the course of different NMR experiments, and then compares the results of model calculations for optimized water polarization with experimental data. It thus demonstrates that NMR spectra of large molecular structures can be improved with the use of paramagnetic spin relaxation agents which selectively enhance the relaxation of water protons, so that a substantial gain in signal-to-noise can be achieved. The presently proposed use of a relaxation agent can also replace the water flip-back pulses when working with structures larger than about 30 kDa. This may be a valid alternative in situations where flip-back pulses are difficult to introduce into the overall experimental scheme, or where they would interfere with other requirements of the NMR experiment. PMID- 16041485 TI - Protein energetic conformational analysis from NMR chemical shifts (PECAN) and its use in determining secondary structural elements. AB - We present an energy model that combines information from the amino acid sequence of a protein and available NMR chemical shifts for the purposes of identifying low energy conformations and determining elements of secondary structure. The model ("PECAN", Protein Energetic Conformational Analysis from NMR chemical shifts) optimizes a combination of sequence information and residue-specific statistical energy function to yield energetic descriptions most favorable to predicting secondary structure. Compared to prior methods for secondary structure determination, PECAN provides increased accuracy and range, particularly in regions of extended structure. Moreover, PECAN uses the energetics to identify residues located at the boundaries between regions of predicted secondary structure that may not fit the stringent secondary structure class definitions. The energy model offers insights into the local energetic patterns that underlie conformational preferences. For example, it shows that the information content for defining secondary structure is localized about a residue and reaches a maximum when two residues on either side are considered. The current release of the PECAN software determines the well-defined regions of secondary structure in novel proteins with assigned chemical shifts with an overall accuracy of 90%, which is close to the practical limit of achievable accuracy in classifying the states. PMID- 16041486 TI - Solution structure of the HMG-box domain in the SSRP1 subunit of FACT. PMID- 16041487 TI - 1H, 13C and 15N assignments for the II-III loop region of the skeletal dyhydropyridine receptor. PMID- 16041488 TI - NMR assignment of the gpU tail protein from lambda bacteriophage. PMID- 16041489 TI - NMR assignment of the periplasmic domain of peptidoglycan-associated lipoprotein (Pal) from Haemophilus influenzae. PMID- 16041490 TI - NMR assignment of the R2 domain of pneumococcal choline binding protein A (CbpA). PMID- 16041491 TI - NMR assignment of the six zinc fingers of MTF-1 in the free and DNA-bound states. PMID- 16041492 TI - NMR assignment of the reduced and oxidized forms of the human ADAP hSH3-1 domain. PMID- 16041494 TI - Region-dependent role of the mucous/glycocalyx layers in insulin permeation across rat small intestinal membrane. AB - The regional difference in the contribution of the mucous/glycocalyx layers in rat small intestine, as a diffusional or enzymatic barrier, to the absorption of insulin was investigated by in vitro studies. The mucous/glycocalyx layers from the duodenum, the jejunum, and the ileum in rat were successfully removed without damaging membrane integrity, by exposing them to a hyaluronidase solution in situ. In an in vitro transport experiment, the apparent permeability coefficient (P(app)) of insulin for the hyaluronidase-pretreated group was significantly increased compared to the PBS-pretreated (control) group in all small intestinal regions, and the P(app) of insulin in both PBS- and hyaluronidase-pretreated groups increased in the following order: duodenum < jejunum < ileum. On the other hand, irrespective of small intestinal regions, the P(app) of FD-4 and of antipyrine, respectively the passive para- and transcellular permeation marker, exhibited no significant differences between PBS- and hyaluronidase-pretreated group. In addition, a significant amount of insulin was degraded in the mucous/glycocalyx layers compartment removed by hyaluronidase pretreatment, and the degradation activity in the mucous/glycocalyx layers showed regional differences in the following order: duodenum > jejunum > ileum. These findings suggest that, irrespective of small intestinal regions, the mucous/glycocalyx layers contributed to insulin permeation predominantly as an enzymatic barrier, and not as a diffusional barrier. Furthermore, the variation of the enzymatic activities in the mucous/glycocalyx layers and in the brush-border membrane would be one factor that accounts for the regional differences in the transport of insulin. PMID- 16041495 TI - The Alu insertion in the CLCN5 gene of a patient with Dent's disease leads to exon 11 skipping. AB - Alu sequences are short, interspersed elements that have generated more than one million copies in the human genome. They propagate by transcription followed by reverse transcription and integration, causing mutations, recombination, and changes in pre-mRNA splicing. We have recently identified a 345-bp long Alu Ya5 element inserted in codon 650 within exon 11 of the chloride channel ClC-5 gene (CLCN5) of a patient with Dent's disease. A microsatellite pedigree analysis indicated that the insertion occurred in the germline of the maternal grandfather. Dent's disease is an X-linked renal tubular disorder characterized by low-molecular-weight proteinuria, hypercalciuria, nephrolithiasis, and nephrocalcinosis. Here, we found, by RT-PCR amplification of RNA extracted from the patient's blood and subsequent DNA sequencing, that the Alu insertion led to an aberrant splicing of the CLCN5 pre-mRNA that skipped exon 11. Using the ESE finder and RESCUE-ESE Web interfaces, we identified two high-score exonic splicing enhancer (ESE) sequences in the site of insertion. The functional significance of these ESE motifs is suggested by our observation that these sequences are highly conserved among mammal CLCN5 genes. Therefore, we suggest that the Alu insertion causes exon skipping by interfering with splicing regulatory elements. The altered splicing would predict a truncated ClC-5 protein that lacks critical domains for sorting and chloride channel function. PMID- 16041496 TI - Genetic diversity and new therapeutic concepts. AB - The differences in medicinal drug responses among individuals had been known for quite some time. Some patients exhibit a life-threatening adverse reaction while others fail to show an expected therapeutic effect. Intermediate responses between the above two extreme cases are also known. In fact, it has been recently reported that approximately 100,000 deaths and more than 2 million hospitalizations annually in the United States are due to properly prescribed medications. This interindividual variability could be due in part to genetically determined characteristics of target genes or drug metabolizing enzymes. This has now been substantiated by a variety of studies. We know that "one size fits all" is not correct. Therefore, the application of pharmacogenetic concepts to clinical practice is an excellent goal in the postgenomic era. The successful completion of the human genome project provided necessary molecular tools, such as high-throughput SNP genotyping, HapMap, and microarray, that can be applied to develop proper therapeutic options for individuals. Recently, there have been considerable scientific, corporate, and policy interest in pharmacotherapy. However, identification of causal variations in a target gene is only a starting point, and the progress in this rapidly developing field is slower than expected. One major drawback could be due to the multigene determinant of drug response that requires a genome-wide screening. Additionally, application of pharmacogenetic knowledge into clinical practice requires a high level of accuracy, precision (risk/benefit ratio), and strict regulations. This is because the pharmacogenetic approach raises several ethical, moral, and legal questions. It is also necessary that both health professionals and the general public must be urgently educated. Despite these limitations, translation of pharmacogenomic data into clinical practice would certainly provide better opportunities to increase the safety and efficacy of medicine in the future. PMID- 16041497 TI - Is the auditory sensory memory sensitive to visual information? AB - The mismatch negativity (MMN) component of auditory event-related brain potentials can be used as a probe to study the representation of sounds in auditory sensory memory (ASM). Yet it has been shown that an auditory MMN can also be elicited by an illusory auditory deviance induced by visual changes. This suggests that some visual information may be encoded in ASM and is accessible to the auditory MMN process. It is not known, however, whether visual information affects ASM representation for any audiovisual event or whether this phenomenon is limited to specific domains in which strong audiovisual illusions occur. To highlight this issue, we have compared the topographies of MMNs elicited by non speech audiovisual stimuli deviating from audiovisual standards on the visual, the auditory, or both dimensions. Contrary to what occurs with audiovisual illusions, each unimodal deviant elicited sensory-specific MMNs, and the MMN to audiovisual deviants included both sensory components. The visual MMN was, however, different from a genuine visual MMN obtained in a visual-only control oddball paradigm, suggesting that auditory and visual information interacts before the MMN process occurs. Furthermore, the MMN to audiovisual deviants was significantly different from the sum of the two sensory-specific MMNs, showing that the processes of visual and auditory change detection are not completely independent. PMID- 16041498 TI - Cutaneous reflexes during rhythmic arm cycling are insensitive to asymmetrical changes in crank length. AB - The neural control of a movement depends upon the motor task performed. To further understand the neural regulation of different variations of the same type of movement, we created three dissimilar bilateral rhythmic arm cycling tasks by unilaterally manipulating crank length (CL). Modulation in the amplitude and sign of cutaneous reflexes was used as an index of neural control. Neurologically intact subjects performed three bilateral cycling trials at approximately 1 Hz with the ipsilateral crank arm at one of three different lengths. Cutaneous reflexes were evoked during each trial with trains (5 x 1.0 ms pulses at 300 Hz) of electrical stimulation delivered to the superficial radial nerve at the ipsilateral wrist. EMG recordings were made bilaterally from muscles acting at the shoulder, elbow, and wrist. Analysis was conducted after phase-averaging contingent upon the timing of stimulation in the movement cycle. CL variation created an asymmetrical cycling pattern and produced significant changes in the range of motion at the ipsilateral shoulder and elbow. Background EMG amplitude in muscles of the contralateral arm generally increased significantly as CL decreased. Therefore at a given phase in the movement cycle, the background EMG was different between the three cycling trials. In contrast, cutaneous reflex amplitudes in muscles of both arms were similar at each phase of the movement cycle between the different CLs trials at both early and middle latencies. This was particularly evident in muscles ipsilateral to nerve stimulation. We suggest that variations of arm cycling that primarily yield significant changes in the amplitude of muscle activity do not require significant task-specific change in neural control. PMID- 16041499 TI - Early adaptations in somatosensory cortex after focal ischemic injury to motor cortex. AB - In response to a lesion, intact regions of cortex in both hemispheres undergo adaptive changes in network function. For example, changes in excitability and intracortical inhibition in primary motor cortex (M1) were reported after lesioning contralateral or ipsilateral brain regions. Close interactions exist between M1 and primary somatosensory cortex (S1) within one hemisphere. Therefore, we hypothesized that lasting modifications would occur in S1 excitability after lesioning ipsilateral M1. Imaging of intrinsic optical signals (IOS, at 570 nm) was used to investigate the evolution of the somatosensory cortical response evoked by contralateral median nerve stimulation during the first hour after a photothrombotic lesion to M1 (caudal motor cortex) of the rat (n=10). Control rats (n=6) received no lesion. Perfusion was monitored by Laser speckle imaging and the extent of the resulting lesion was determined histologically. Control animals did not show evidence for reduced perfusion, infarction, or changes in IOS. M1 infarction led to a significant increase in evoked response amplitude, duration, and area of activation, and a shortening of latencies. These parameters reached a plateau around 50 min after ischemia. These results indicate S1 hyperexcitability after M1 injury. Whether these adaptations contribute to functional deficits or play a role in recovery, remains to be determined. PMID- 16041501 TI - Vestibulo-collic reflex (VCR) in mice. AB - The vestibulo-collic reflex (VCR) attempts to stabilize head position in space during motion of the body. Similar to the better-studied vestibulo-ocular reflex, the VCR is subserved by relatively direct, as well as indirect pathways linking vestibular nerve activity to cervical motor neurons. We measured the VCR using an electromagnetic technique often employed to measure eye movements; we attached a loop of wire (head coil) to an animal's head using an adhesive; then the animal was gently restrained with its head free to move within an electromagnetic field, and was subjected to sinusoidal (0.5-3 Hz) or abrupt angular acceleration (peak velocity approximately 200 degrees/s). Head rotation opposite in direction to body rotation was assumed to be driven by the VCR. To confirm that the compensatory head movements were in fact vestibular in origin, we plugged the horizontal canal unilaterally and then retested the animals 2, 8 and 15 days after the lesion. Two days after surgery, the putative VCR was almost absent in response to abrupt or sinusoidal rotations. Recovery commenced by day 8 and was nearly complete by day 15. We conclude that the compensatory head movements are vestibular in origin produced by the VCR. Similar to other species, there are robust compensatory mechanisms that restore the VCR following peripheral lesions. PMID- 16041500 TI - Deceleration affects anticipatory and reactive components of triggered postural responses. AB - Understanding the physiological and psychological factors that contribute to healthy and pathological balance control in man has been made difficult by the confounding effects of the perturbations used to test balance reactions. The present study examined how postural responses were influenced by the acceleration deceleration interval of an unexpected horizontal translation. Twelve adult males maintained balance during unexpected forward and backward surface translations with two different acceleration-deceleration intervals and presentation orders (serial or random). "SHORT" perturbations consisted of an initial acceleration (peak acceleration 1.3 m s(-2); duration 300 ms) followed 100 ms later by a deceleration. "LONG" perturbations had the same acceleration as SHORT perturbations, followed by a 2-s interval of constant velocity before deceleration. Surface and intra-muscular electromyography (EMG) from the leg, trunk, and shoulder muscles were recorded along with motion and force plate data. LONG perturbations induced larger trunk displacements compared to SHORT perturbations when presented randomly and larger EMG responses in proximal and distal muscles during later (500-800 ms) response intervals. During SHORT perturbations, activity in some antagonist muscles was found to be associated with deceleration and not the initial acceleration of the support surface. When predictable, SHORT perturbations facilitated the use of anticipatory mechanisms to attenuate early (100-400 ms) EMG response amplitudes, ankle torque change and trunk displacement. In contrast, LONG perturbations, without an early deceleration effect, did not facilitate anticipatory changes when presented in a predictable order. Therefore, perturbations with a short acceleration deceleration interval can influence triggered postural responses through reactive effects and, when predictable with repeated exposure, through anticipatory mechanisms. PMID- 16041502 TI - Mucositis prevention by improved dental care in acute leukemia patients. AB - GOALS OF WORK: The aim of the present study was to evaluate the effects of the intensive dental care protocol in preventing oral complications in acute leukemia patients. PATIENTS AND METHODS: Thirty-four patients hospitalized for induction remission therapy for acute leukemia were randomly assigned to one of two groups, whether to receive intensive dental care protocol or not. The intensive dental care group of patients received dental treatment and plaque and calculus removal prior to chemotherapy and supervised oral hygiene measures during chemotherapy. The limited dental care group of patients did not receive prechemotherapy dental care. Groups were comparable in age, sex, and antineoplastic treatment received. Patients were examined after admission to the hospital; at the initiation of the chemotherapy; and 7, 14, 21, and 28 days after initiation of therapy. Positive data about subjective difficulties were taken by anamnesis. Oral hygiene index (OHI) and gingival index (GI) were used to assess the periodontal status of the patients. The severity of mucositis was evaluated according to WHO classification. MAIN RESULTS: The results of this study pointed out lower mean values of GI and lower mean values of mucositis score in the intensive dental care group of patients during the whole period of examination. Although the differences in mean values were not statistically significant on most of the examination days, intensive dental care group of patients developed less severe and less painful oral complications compared to the limited dental care group of patients. CONCLUSION: We conclude that proper dental care and preventive measures both before and during chemotherapy can be beneficial to these patients. PMID- 16041503 TI - Late sequelae of radiotherapy in adults. AB - Around 3-5 million cancer patients receive radiotherapy worldwide every year. Improved cure rates have led to physicians being confronted with a range of late radiation sequelae, broadly of two types-late effects on normal tissues (LENT) and radiation-induced second malignancies (RISM). These late changes affect the function and quality of life of cancer survivors, increase with increasing radiation dose and show a relationship with time. Overall risks for LENT and RISM are 5-10% and 2-10%, respectively, and the potential benefits of radiotherapy outweigh these risks. PMID- 16041504 TI - [Salivary duct carcinoma]. AB - This tutorial focuses on salivary duct carcinoma (SDC), a rare, high grade neoplasm mainly of major salivary glands. The clinical course of these tumors is characterised by extended local disease, early distant metastasis, and poor outcome. The morphology of SDC is reminiscent of breast ductal carcinomas and may occasionally cause diagnostic problems. In spite of mimicry with ductal carcinoma in situ of the breast and an in situ component, that is evident in most tumors by immunohistology with antibodies directed against high molecular weight cytokeratins (Ck), SDC is always an invasive carcinoma. By immunohistology, most tumors show reactivity with antibodies directed against Ck 7, Ck 8/18 and Ck 19 whereas a morphologically indistinguishable subgroup expresses Ck 5/6 in tumor cells in addition to residual basal epithelia. Carcinoembryonic antigen, GCDFP-15 and androgen receptor are other helpful markers in routine diagnosis of SDC. Prostate-specific antigen is detectable in some cases. Abnormal p53 expression seems to indicate an adverse prognosis. Expression of c-erbB2, the over expression of which is associated with a poor prognosis, may form the basis for a targeted therapeutic approach for selected cases of SDC. PMID- 16041505 TI - Biotechnological production and application of vitamin E: current state and prospects. AB - Tocochromanols (tocopherols and tocotrienols) are important lipophilic antioxidants for animals and humans. Their biological activity is expressed as vitamin E activity. This article describes the current need for vitamin E production, and compares different strategies to engineer the vitamin E content in photosynthetic bacteria and plants, with a focus on oilseed as target tissues. The current status of biotechnological advances in tocochromanol pathway engineering is summarized, and current limitations in our understanding of the tocochromanol biosynthetic pathway are discussed. PMID- 16041506 TI - [Acute dyspnoea]. AB - Asthma, chronic obstructive pulmonary diseases, interstitial lung diseases and myocardial dysfunction are the leading diagnoses of patients with dyspnoea. Acute dyspnoea appears suddenly or within a few hours and is caused by a relatively limited number of conditions some of which can be life-threatening. Therefore, it is of utmost importance to identify the cause of dyspnoea immediately. Once found, the disorder is treated according to the guidelines for the underlying disease. PMID- 16041508 TI - Surgical emergency for inflammatory bowel disease. PMID- 16041507 TI - A comparative study of the expression of Wnt-1, WISP-1, survivin and cyclin-D1 in colorectal carcinoma. AB - BACKGROUND AND AIMS: It is well accepted that activation of Wnt signalling occurs in colorectal carcinoma (CRC), but the correlation amongst the various proteins involved in primary tumours are still unclear. The expression of the inducer of this pathway, Wnt-1, and the downstream effectors, WISP-1, cyclin-D1 and survivin proteins, was compared in a series of CRC tissues with the apparently normal adjacent tissues to determine the relationship of these proteins. PATIENTS AND METHODS: Formalin-fixed, paraffin-embedded tissue samples of 47 CRCs surgically resected at the Kuala Lumpur Hospital (KLH) between 1999 and 2000 were used. Immunohistochemical staining with monoclonal antibodies against cyclin-D1 and survivin and polyclonal antibodies against Wnt-1 and WISP-1 was performed. Results of immunohistochemistry were analysed for correlation between biomolecules and histopathological data of the patients. RESULTS: Of the 47 CRCs, 26 (55.3%), 15 (31.9%), 5 (10.6%) and 28 (59.6%) of the tumours exhibited positivity for Wnt-1, WISP-1, cyclin D1 and survivin, respectively. A lower percentage of the 40 apparently normal adjacent tissues were found to be positive for Wnt-1 (7, 17.5%), WISP-1 (+/-5, 12.5%) and survivin (13, 32.5%), but cyclin D1 was not detected in any of them. Interestingly, the total scores of Wnt-1, WISP-1 and survivin were significantly higher in CRC tissues (p=0.001, 0.034 and 0.044, respectively). Using the Spearman rank correlation test, a positive linear relationship was found between total Wnt-1 score with total WISP-1 score (rho=0.319, p=0.003) and total survivin score (rho=0.609, p=or<0.001). The expression of WISP-1 in the CRC tissues was found to be positively correlated with patients older than 60 years old (p=0.011). In addition, nuclear cyclin-D1 expression was found to be associated with poorly differentiated CRC tissues (p<0.001, Table 5) and right-sided CRC tumour (p=0.019, Table 6). Total WISP-1 score was associated with well-differentiated CRC tissues (p=0.029). CONCLUSIONS: Overexpression and interplay between Wnt-1, WISP-1, survivin and cyclin-D1 may play a role in tumorigenesis, possibly by promoting cell cycle checkpoint progression, accelerating cell growth and inhibiting apoptosis. Our data may provide useful information towards the search for potent therapeutic targets towards the development of novel treatment strategies for CRC. PMID- 16041509 TI - Automatic motor cortex activation for natural as compared to awkward grips of a manipulable object. AB - It has been suggested that, relative to natural objects, man-made object representations in the brain are more specifically defined by functional properties that reflect how an object is used and/or what it is used for (Warrington and Shallice 1984). We recorded 123-channel event-related potentials (ERP) in healthy participants during a mental rotation task involving a manipulable (hammer) and a non-manipulable (church) object. Both stimuli had standard and mirror-image versions rotated in four different orientations, resulting for the manipulable object in some natural and some awkward grips. Using spatial cluster analysis, time periods were determined during which the ERP maps differed between stimulus conditions. Specific maps appeared for natural versus awkward grips with the manipulable object at a very early stage (60-116 ms) as well as during a later stage (180-280 ms). Source estimations derived from the topographic data indicated that during the second time window the left motor cortex was significantly activated in the case of natural grips. We argue that the motor programs that are semantically associated with the object are automatically activated when it is presented in graspable orientations. PMID- 16041510 TI - Hand preshaping in Parkinson's disease: effects of visual feedback and medication state. AB - Previous studies in our laboratory examining pointing and reach-to-grasp movements of Parkinson's disease patients (PDPs) have found that PDPs exhibit specific deficits in movement coordination and in the sensorimotor transformations required to accurately guide movements. We have identified a particular difficulty in matching unseen limb position, sensed by proprioception, with a visible target. In the present work, we further explored aspects of complex sensorimotor transformation and motor coordination using a reach-to-grasp task in which object shape, visual feedback, and dopaminergic medication were varied. Normal performance in this task requires coordinated generation of appropriate reach, to bring the hand to the target, and differentiated grasp, to preshape the hand congruent with object form. In Experiment 1, we tested PDPs in the off-medication state. To examine the dependence of subjects on visual feedback and their ability to implement intermodal sensory integration, we required them to reach and grasp the target objects in three conditions: (1) Full Vision, (2) Object Vision with only the target object visible and, (3) No Vision with neither the moving arm nor the target object visible. PDPs exhibited two types of deficits. First, in all conditions, they demonstrated a generalized slowing of movement or bradykinesia. We consider this an intensive deficit, since it involves largely a modulation of the gain of specific task parameters: in this case, velocity of movement. Second, they were less able than controls to extract critical proprioceptive information and integrate it with vision in order to coordinate the reach and grasp components of movement. These deficits which involve the coordination of different inputs and motor components, we classify as coordinative deficits. As in our previous work, the PDPs' deficits were most marked when they were required to use proprioception to guide their hand to a visible target (Object Vision condition). But even in the full-vision condition, their performance only became fully accurate when both the target and effector (hand) were simultaneously visible. In Experiment 2, PDPs were tested on their dopaminergic replacement therapy. Dopaminergic treatment significantly ameliorated the bradykinesia of the PDPs, but produced no changes in the hand preshaping deficiencies of PDPs. These results suggest that adequate treatment of the PDPs may more readily compensate for intensive, than coordinative deficits, since the latter are likely to depend on specific and time-dependent neural interdependencies that are unlikely to be remediated simply by increasing the gain of a pathway. PMID- 16041511 TI - Habituation reduces novelty-induced FOS expression in the striatum and cingulate cortex. AB - Exposure to novel environments is known to induce c-fos expression in the cingulate cortex and dorsomedial striatum. This study examined the effects of 1, 3, and 7 days of exposure to sequential exposure to three novel environments. It was hypothesized that the ability of novel environment shuttling (NES) would induce c-fos expression as indicated by cells displaying FOS-like immunoreactivity (FLI) in the striatum and cingulate cortex which would decrease with repeated exposure. NES elicited FLI in both the cingulate cortex and striatum when compared to home-cage controls and this effect was attenuated with repeated exposure. Behavioral measures of activity decreased with repeated exposure suggesting that the decrease in FLI may be due to either a reduction in motor activity or increasing familiarity with the environments. PMID- 16041512 TI - The effects of landmarks on the performance of delayed and real-time pointing movements. AB - Converging lines of evidence suggest that the presence of non-target landmarks affects the performance of delayed target-directed movements (e.g., Diedrichsen et al. 2004; Sheth and Shimojo 2004). In the present experiment, we examined the effects of non-target landmarks on the accuracy and precision of delayed and immediate target-directed pointing movements. In our experiment, the landmarks were present just prior to and during the presentation of the target; they were never present during the execution of the movement. Absolute errors were significantly reduced when the landmarks were available during target presentation for both delayed and immediate action conditions. In contrast, the presence of landmarks improved the precision of delayed but not immediate movements (as indexed by the variable error). The locus of this "landmark benefit" appears to be in the encoding of target position since landmarks were never available after target offset. We suggest that, when available, information provided by landmarks is used to improve the accuracy of the estimation of target location. Since the targets were presented for only 100 ms, it is apparent that the spatial information available from landmarks can be quite rapidly used to estimate target position. Further, with respect to the precision of movements, we suggest that the presence of landmarks serves to improve the stability of the estimation of target position. This particular reliance on landmark information becomes more critical as the movement is delayed. PMID- 16041513 TI - What is the coordinate frame utilized for the generation of express saccades in monkeys? AB - The latencies of saccades to suddenly appearing eccentric targets can have a bimodal distribution, with an early, express peak, and a late, regular peak (Fischer and Boch 1983, Brain Res 260: 21-26). Express saccades usually are a product of learning. The purpose of this study was to determine whether this learning is specific to the relative position of the target in space, the orbital position of the eye, or the vector of the saccade to be produced. Further, it was asked whether and how the frequency with which express saccades are generated is influenced by the immediately preceding saccadic vector and the familiarity of the targets. To this end, rhesus monkeys were trained to make saccadic eye movements to single targets and to two sequential targets that appeared at various positions relative to the head, relative to the initial fixation spot and relative to each other. The results show that the frequency with which express saccades are generated is determined by the saccadic vector that has to be generated and not by the relative position of a target in space, the orbital position of the eye, the immediately preceding saccadic vector, or the familiarity of the targets. PMID- 16041514 TI - Dynamics and directionality of the vestibulo-collic reflex (VCR) in mice. AB - The vestibulo-collic reflex (VCR) stabilizes the head in space by excitation of neck muscles that oppose head rotation. Recently, the mouse vestibulo-ocular reflex (VOR) has been characterized so that genetic manipulations of the vestibular system can be examined. We have characterized the dynamics and directionality of the VCR in mice restrained at the neck so that studies of vestibular system genetics may include comparisons to normal VCR in addition to VOR. Head rotations were measured in darkness with a three-dimensional search coil system during whole body rotations. The VCR in four C57BL/6 mice was present in pitch, roll, and yaw directions with an overall average gain of 0.28. Phase was accurately compensatory to oppose head rotation across a wide range of frequencies from 0.02 Hz to 2.0 Hz. Compensatory head rotations were greatest in the direction opposing the applied stimulus and weak or absent in other directions. Constant velocity rotations about horizontal axes elicited head velocity modulation and bias similar to that observed in the VOR. We conclude that the VCR of mice is similar to that in other mammals. PMID- 16041515 TI - Construction and one-step purification of Bacillus kaustophilus leucine aminopeptidase fused to the starch-binding domain of Bacillus sp. strain TS-23 alpha-amylase. AB - The starch-binding domain of Bacillus sp. strain TS-23 alpha-amylase was introduced into the C-terminal end of Bacillus kaustophilus leucine aminopeptidase (BkLAP) to generate a chimeric enzyme (BkLAPsbd) with raw-starch binding activity. BkLAPsbd, with an apparent molecular mass of approximately 65 kDa, was overexpressed in Escherichia coli M15 cells and purified to homogeneity by nickel-chelate chromatography. Native PAGE and chromatographic analyses revealed that the purified fusion protein has a hexameric structure. The half life for BkLAPsbd was 12 min at 70 degrees C, while less than 20% of wild-type enzyme activity retained at the same heating condition. Compared with the wild type enzyme, the 60% decrease in the catalytic efficiency of BkLAPsbd was due to a 91% increase in K (m) value. Starch-binding assays showed that the K (d) and B (max) values for the fusion enzyme were 2.3 microM and 0.35 micromol/g, respectively. The adsorption of the crude BkLAPsbd onto raw starch was affected by starch concentration, pH, and temperature. The adsorbed enzyme could be eluted from the adsorbent by 2% soluble starch in 20 mM Tris-HCl buffer (pH 8.0). About 49% of BkLAPsbd in the crude extract was recovered through one adsorption-elution cycle with a purification of 11.4-fold. PMID- 16041516 TI - Quality assurance challenge 2. AB - We would like to invite you to participate in the Analytical Challenge, which is a series of puzzles designed to entertain and challenge our readers. The special ABC feature 'Analytical Challenge' has established itself as a truly unique quiz series, with a new scientific puzzle published every second month. Readers can access the complete collection of published problems with their solutions on the ABC homepage at www.springeronline.com/journal/00216. Test your knowledge and tease your wits in diverse areas of analytical and bioanalytical chemistry by viewing this collection. Quality assurance is the topic of the present challenge. And please note: there is a prize to be won (a Springer book from our catalogue up to a value of euros 75,-). Please read on... PMID- 16041517 TI - Gaseous nitrogen oxide repressed benzo[a]pyrene-induced human lung fibroblast cell apoptosis via inhibiting JNK1 signals. AB - Benzo[a]pyrene (B[a]P) is present in environmental pollution and cigarette smoke. B[a]P has been shown to induce apoptosis in hepatoma cells, human B cells, human ectocervical cells, macrophages, and rat lungs. Nitrogen oxides (NOx) are the other important indoor and outdoor air pollutants. Many studies have indicated that NO gas causes lung tissue damage both by its oxidative properties and free radicals. In our previous study we demonstrated that NO gas induced proliferation of human lung fibroblast MRC-5 cells. In this study we showed that NO gas inhibits B[a]P-induced MRC-5 cells apoptosis by cell cycle analysis. Western blot data revealed that NO gas increased the expressions of anti-apoptosis proteins (Bcl-2 and Mcl-1) and decreased the expression of apoptosis proteins (Bax, t-Bid, cytochrome c, FasL, and caspases) after B[a]P treatment. We further clarified that B[a]P-induced MRC-5 cell apoptosis via JNK1/FasL and JNK1/p53 signals. In conclusion, NO gas inhibited B[a]P-induced MRC-5 cells apoptosis via inhibition of JNK1 apoptosis pathway and induction of Bcl-2 and Mcl-1 anti-apoptosis pathway. PMID- 16041518 TI - [Munchausen syndrome and chronic otitis media]. AB - In this case report an artificial disorder with symptoms of chronic otitis media and progressive hearing loss is described. This represents a rare case of Munchausen syndrome. The difficulties in diagnosis and therapy are shown by the development of the disease, produced by self damaging activities. PMID- 16041519 TI - Communication of end-of-life decisions in European intensive care units. AB - OBJECTIVE: To examine end-of-life (EOL) practices in European ICUs: who makes these decisions, how they are made, communication of these decisions and questions on communication between the physicians, nurses, patients and families. DESIGN: Data collected prospectively on EOL decisions facilitated by a questionnaire including EOL decision categories, geographical regions, mental competency, information about patient wishes, and discussions with patients, families and health care professionals. SETTING: 37 European ICUs in 17 countries. PATIENTS: ICU physicians collected data on 4,248 patients. RESULTS: 95% of patients lacked decision making capacity at the time of EOL decision and patient's wishes were known in only 20% of cases. EOL decisions were discussed with the family in 68% of cases. Physicians reported having more information about patients' wishes and discussions in the northern countries (31%, 88%) than central (16%, 70%) or southern (13%, 48%) countries. The family was more often told (88%) than asked (38%) about EOL decisions. Physicians' reasons for not discussing EOL care with the family included the fact that the patient was unresponsive to therapy (39%), the family was unavailable (28%), and the family was thought not to understand (25%). CONCLUSIONS: ICU patients typically lack decision-making capacity, and physicians know patients' wishes in only 20% of EOL decisions. There were regional differences in discussions of EOL decisions with families and other physicians. In European ICUs there seems to be a need to improve communication. PMID- 16041520 TI - Successful use of parenteral ivermectin in an immunosuppressed patient with disseminated strongyloidiasis and septic shock. PMID- 16041521 TI - Elevated carboxyhemoglobin associated with sodium nitroprusside treatment. AB - OBJECTIVE: To report four cases of carboxyhemoglobinemia associated with high doses of sodium nitroprusside after cardiac transplant in children. PATIENTS: Four children in the pediatric care unit of a university hospital aged 6 months-4 years. Carboxyhemoglonemia developed at levels of 5.5-7.7% in patients receiving high doses of sodium nitroprusside (7-16 microg/kg per minute and no other medication that could caused elevated carboxyhemoglobin). One patient died, and three recovered with no sequelae after discontinuation of sodium nitroprusside. CONCLUSIONS: High doses of sodium nitroprusside can induce carboxyhemoglobinemia in children after heart transplant, probably by inducing hemeoxygenase, with no other secondary effects. PMID- 16041522 TI - Effects of short vs. prolonged mechanical ventilation on antioxidant systems in piglet diaphragm. AB - OBJECTIVE: Prolonged controlled mechanical ventilation (MV) is known to induce diaphragmatic oxidative stress that seems to be an important factor reducing force-generating capacity. To better understand the cellular mechanisms involved, this work examined the effect of short vs. prolonged MV on antioxidant defense in the diaphragm. DESIGN AND SETTING: Prospective, randomized, controlled animal study in a university laboratory. METHODS: Eleven piglets (15-20 kg) were assigned to one of two groups: a long-MV group (n=6) ventilated for 3 days or a short-MV group (n=5) ventilated for 3 h. Force frequency curves of the transdiaphragmatic pressure (Pdi) were obtained in vivo by phrenic nerve pacing. Oxidative stress was evaluated by thiobarbituric reactive substance (TBARs) content and the enzymatic antioxidant activity of both superoxide dismutase (SOD) and glutathione peroxidase (GPx) in samples of diaphragm. RESULTS: Pdi decreased in the long-MV group by 30-35% over the 3 days at all frequencies compared to the short-MV group. Diaphragm TBARs content was significantly higher and SOD activity lower in long-MV animals than in short-MV animals after 72 h. GPx activity tended to be lower in diaphragms from long-MV animals, but this difference was not significant. CONCLUSIONS: This study shows that 3 days of MV in piglets is associated with a decrease in antioxidant activity which could emphasize oxidative stress and both contribute to the diaphragm dysfunction caused by MV. PMID- 16041523 TI - Diaphragm antioxidant system in controlled mechanical ventilation in piglets: short term vs. prolonged mechanical ventilation response. PMID- 16041524 TI - Evaluation of the Glucometer Elite XL device for screening for neonatal hypoglycaemia. AB - To prevent persistent neurodevelopment and physical growth deficits in neonatal care, it is mandatory to determine blood glucose levels as quickly and precisely as possible, preferably using micro-methods. However, most commercially available instruments have not been validated and approved for this purpose. The aim of this study was to validate the Glucometer Elite XL, a newly developed device for point-of-care testing (POCT). In samples from 869 newborn infants, glucose levels were simultaneously measured by the Glucometer Elite XL in whole blood and by an accepted clinical laboratory method in haemolysed blood using the ECA 2000 device. An acceptable method agreement was found between the POCT and the ECA 2000 method (mean difference 0.013 mmol/l, SD 0.69). As determined by regression analysis (Passing-Bablok), the slope was 1.086 with a y-intercept of -0.4 mmol/l ( r =0.959, P <0.05). The differences between measurement pairs of both assays versus the haematocrit were negligible. With a cut-off for hypoglycaemia at 2.6 mmol/l glucose in haemolysed blood, the sensitivity of the POCT device was 0.63 and specificity was 0.98. Raising the cut-off of the Glucometer Elite XL to 3.2 mmol/l, the sensitivity and specificity incremented to 1.0 and 0.89, respectively. CONCLUSION: The Glucometer Elite XL instrument can be recommended for point-of-care blood glucose measurement in newborn infants if its character as a screening method is taken into account. To compensate deviating results, we advise to shift its cut-off for hypoglycaemia recognition to a safe threshold of 3.2 mmol/l. However, hypoglycaemia has to be confirmed by a valid glucose measurement in the clinical laboratory. PMID- 16041525 TI - Pilomatricoma in childhood: a retrospective study from three European paediatric centres. AB - Pilomatricoma is characterised as a common, slowly growing benign cutaneous tumour that appears generally within the first decades of life. The clinical diagnosis is frequently missed, especially by the paediatrician unfamiliar with these tumours. We present the experience gained in three European tertiary care paediatric centres with the treatment of pilomatricoma and also current data on the aetiology, clinical presentation and management. A retrospective study was carried out in 83 patients suspected for pilomatricoma during a 7-year period (1996-2002) at the departments of Paediatric Surgery of the Children's University Hospital "Federico II", Naples, Hospital "San Bortolo", Vicenza and "Aghia Sophia" Children's Hospital, Athens. The age range was from 10 months to 17 years, median age 8 years. All patients were treated by surgical excision and all specimens were examined by histopathological assessment. The follow-up varied from 5 months to 6 years. The correct diagnosis was made preoperatively in 68 patients (82%). The female/male ratio was 2:1. The sites of occurrence were the head (47.5%), especially in the periorbital region, the neck (9%), the upper limbs (35.5%), the inferior limbs (4%) and the thorax (4%). Each patient exhibited a single pilomatricoma except for two patients who had multiple lesions (2.4%). One of them had Steinert disease (myotonic dystrophy). No recurrences were observed during the follow-up period. CONCLUSION: Pilomatricoma is one of the most common cutaneous adnexal neoplasms in children. Surgical excision including clear margins and its overlying skin in most cases is the treatment of choice. The recurrence as well as malignant evolution is rare. PMID- 16041526 TI - Temporomandibular joint destruction in mucolipidosis type III necessitating gastrostomy insertion. AB - Mucolipidosis III is a genetically heterogeneous lysosomal disorder characterised by progressive symptoms and signs, the commonest being skeletal pain due to bony destruction. We describe a patient who developed severe destruction of the temporomandibular joints leading to difficulties with speech and feeding, necessitating gastrostomy insertion. CONCLUSION: Temporomandibular joint involvement has not been previously reported in mucolipidosis III. PMID- 16041527 TI - Comprehensive MR angiography of the lower limbs: a hybrid dual-bolus approach including the pedal arteries. AB - The purpose of this study was to include the pedal vasculature into the coverage of peripheral multistation magnetic resonance angiography (3DceMRA). A total of 216 patients suffering from peripheral vascular disease were examined with a modified hybrid dual-bolus technique. The cruropedal arteries were acquired first with two sagittal slabs and time-resolved 3D sequences. Then the aortofemoral vessels were visualized using the bolus-chase technique and a second contrast injection. Interventional procedures were performed in 104 patients, and in 69 of those, the cruropedal vessels were also examined with digital subtraction angiography (iaDSA). Using 3DceMRA, the cruropedal arteries were displayed with both excellent and good quality in 95% (205/216 cases), and without any venous overlay in 94% (203/216 cases). The aortofemoral vessels were not jeopardized by the first contrast injection. With iaDSA as the standard of reference, observed sensitivity of 3DceMRA was found in ranges from 80% (29%, 99%) to 100% (86%, 100%) for assessing significant stenoses, and observed specificity ranged between 93% [80%, 98%] and 100% (82%, 100%). In conclusion, hybrid dual-bolus 3DceMRA significantly reduces the limitations of standard single-bolus 3DceMRA in anatomic coverage and temporal resolution of the cruropedal arteries, thus providing high-quality images of the entire peripheral vasculature. PMID- 16041528 TI - 3D CT protocol in the assessment of the esophageal neoplastic lesions: can it improve TNM staging? AB - The purpose of this prospective observational study was the evaluation of the usefulness of MPR reconstructions and virtual endoscopy in the study of the esophageal carcinoma. Thirty-nine patients with esophageal cancer proved by means of endoscopy, underwent preoperative TNM staging with dynamic CT of the chest and abdomen with the aid of 3D rendering. Twenty-six patients underwent surgery, and the CT results were compared with histopathologic findings. In staging the T parameter, the CT with 3D reconstructions and virtual endoscopy, showed a sensitivity of 92% and an accuracy of 88%. In staging lymph nodes, the sensitivity in our study was 85%, the specificity 58%, and the accuracy 69%. Our protocol of the study of the esophageal cancer with 3D CT and virtual endoscopy, demonstrated a high concordance with the surgical and pathologic findings. The 3D reconstructed images were very helpful to the surgeons regarding preoperative planning. We performed an observational enquiry, and although this was a small study, it has, however, confirmed that the 3D imaging of the esophagus represents a valuable advantage to conventional imaging. Further studies with a larger number of patients are needed to prove its superiority to traditional CT imaging of the esophagus. PMID- 16041529 TI - Value of high spatial and high temporal resolution magnetic resonance angiography for differentiation between idiopathic and thromboembolic pulmonary hypertension: initial results. AB - Differentiation between different forms of pulmonary hypertension (PH) is essential for correct disease management. The goal of this study was to elucidate the clinical impact of high spatial resolution MR angiography (SR-MRA) and time resolved MRA (TR-MRA) to differentiate between patients with chronic thromboembolic PH (CTEPH) and idiopathic pulmonary arterial hypertension (IPAH). Ten PH patients and five volunteers were examined. Twenty TR-MRA data sets (TA 1.5 s) and SR-MRA (TA 23 s) were acquired. TR-MRA data sets were subtracted as angiography and perfusion images. Evaluation comprised analysis of vascular pathologies on a segmental basis, detection of perfusion defects, and bronchial arteries by two readers in consensus. Technical evaluation comprised evaluation of image quality, signal-to-noise ratio (SNR) measurements, and contrast-media passage time. Visualization of the pulmonary arteries was possible down to a subsegmental (SR-MRA) and to a segmental (TR-MRA) level. SR-MRA outperformed TR MRA in direct visualization of intravascular changes. Patients with IPAH predominantly showed tortuous pulmonary arteries while in CTEPH wall irregularities and abnormal proximal-to-distal tapering was found. Perfusion images showed a diffuse pattern in IPAH and focal defects in CTEPH. TR-MRA and SR MRA resulted in the same final diagnosis. Both MRA techniques allowed for differentiation between IPAH and CTEPH. Therefore, TR-MRA can be used in the clinical setting, especially in dyspneic patients. PMID- 16041530 TI - Activation of heterotrimeric G-proteins independent of a G-protein coupled receptor and the implications for signal processing. AB - Heterotrimeric G-proteins are key transducers for signal transfer from outside the cell, mediating signals emanating from cell-surface G-protein coupled receptors (GPCR). Many, if not all, subtypes of heterotrimeric G-proteins are also regulated by accessory proteins that influence guanine nucleotide binding, guanosine triphosphate (GTP) hydrolysis, or subunit interactions. One subgroup of such accessory proteins (activators of G-protein signaling; AGS proteins) refer to a functionally defined group of proteins that activate selected G-protein signaring systems in the absence of classical G-protein coupled receptors. AGS and related proteins provide unexpected insights into the regulation of the G protein activation-deactivation cycle. Different AGS proteins function as guanine nucleotide exchange factors or guanine nucleotide dissociation inhibitors and may also influence subunit interactions by interaction with GBgamma. These proteins play important roles in the generation or positioning of signaling complexes and of the regulation of GPCR signaling, and as alternative binding partners for G protein subunits. Perhaps of even broader impact is the discovery that AGS proteins provide a foundation for the concept that heterotrimeric G-protein subunits are processing signals within the cell involving intrinsic cues that do not involve the classical signal input from a cell surface GPCR. PMID- 16041531 TI - Gemcitabine, vinorelbine and prednisone for refractory or relapsed aggressive lymphoma, results of a phase II single center study. AB - The optimum therapy for patients with relapsed or refractory aggressive non Hodgkin's lymphomas (NHL) not qualifying for platinum-based and/or high-dose chemotherapy is not known. We conducted a prospective phase II study evaluating a regimen consisting of gemcitabine (1 g/m2, days 1 and 8), vinorelbine (30 mg/m2, days 1 and 8) and prednisone (100 mg/day, days 1-8) (GVP) given every 21 days. Fifteen patients with a median age of 68 years and a median of three previous therapies were enrolled. Diagnoses included B lymphoblastic (n=1), diffuse large B cell (n=10), anaplastic large T cell (n=2) and peripheral T-cell NHL (n=2). The median international prognostic index score was 3 (six patients with a score of 4 or 5). Five patients achieved a complete remission and three patients a partial remission. The median overall survival was 13.8 months, and the median time to next treatment was 4.4 months. Haematological toxicities of World Health Organisation grades 3/4 were leucopenia in 58%, thrombocytopenia in 33% and anaemia in 17% of all courses. Three patients had grade 3 infections. There was no treatment-related mortality. GVP shows substantial activity in poor prognosis relapsed or refractory aggressive lymphomas and is generally well tolerated, but haematological toxicity is dose limiting. PMID- 16041532 TI - Apoptosis: mechanisms and relevance in cancer. AB - Apoptosis or programmed cell death is a process with typical morphological characteristics including plasma membrane blebbing, cell shrinkage, chromatin condensation and fragmentation. A family of cystein-dependent aspartate-directed proteases, called caspases, is responsible for the proteolytic cleavage of cellular proteins leading to the characteristic apoptotic features, e.g. cleavage of caspase-activated DNase resulting in internucleosomal DNA fragmentation. Currently, two pathways for activating caspases have been studied in detail. One starts with ligation of a death ligand to its transmembrane death receptor, followed by recruitment and activation of caspases in the death-inducing signalling complex. The second pathway involves the participation of mitochondria, which release caspase-activating proteins into the cytosol, thereby forming the apoptosome where caspases will bind and become activated. In addition, two other apoptotic pathways are emerging: endoplasmic reticulum stress induced apoptosis and caspase-independent apoptosis. Naturally occurring cell death plays a critical role in many normal processes like foetal development and tissue homeostasis. Dysregulation of apoptosis contributes to many diseases, including cancer. On the other hand, apoptosis-regulating proteins also provide targets for drug discovery and new approaches to the treatment of cancer. PMID- 16041533 TI - Viscosity-mediated motion coupling between pairs of trichobothria on the leg of the spider Cupiennius salei. AB - Arachnids and insects use long, thin hairs as motion sensors to detect signals contained in the movement of the surrounding air. These hairs often form groups with a small spacing of tens to hundreds of micrometers between them. For air oscillation frequencies of biological interest, the potential exists for viscosity-mediated coupling among hairs in a group affecting their response characteristics. Even a small diameter hair can, in principle, affect the flow field around it and the dynamics of the hairs in its neighborhood. The viscosity mediated coupling between a pair of hairs is investigated here both experimentally and theoretically. The conditions for the existence of the coupling effect, and its magnitude as a function of relevant parameters, are determined. In the range of biologically relevant frequencies (30-300 Hz), viscous coupling between pairs of hairs is only very small in the case of the spider Cupiennius salei. Theoretical analysis points to the relatively large spacing between hairs (20 to 50 hair diameters) and the tuning of the hairs to the above-mentioned frequencies to explain the practical absence of coupling. PMID- 16041534 TI - Modulation of memory and visuospatial processes by biperiden and rivastigmine in elderly healthy subjects. AB - RATIONALE: The central cholinergic system is implicated in cognitive functioning. The dysfunction of this system is expressed in many diseases like Alzheimer's disease, dementia of Lewy body, Parkinson's disease and vascular dementia. In recent animal studies, it was found that selective cholinergic modulation affects visuospatial processes even more than memory function. OBJECTIVE: In the current study, we tried to replicate those findings. In order to investigate the acute effects of cholinergic drugs on memory and visuospatial functions, a selective anticholinergic drug, biperiden, was compared to a selective acetylcholinesterase inhibiting drug, rivastigmine, in healthy elderly subjects. METHODS: A double blind, placebo-controlled, randomised, cross-over study was performed in 16 healthy, elderly volunteers (eight men, eight women; mean age 66.1, SD 4.46 years). All subjects received biperiden (2 mg), rivastigmine (3 mg) and placebo with an interval of 7 days between them. Testing took place 1 h after drug intake (which was around Tmax for both drugs). Subjects were presented with tests for episodic memory (wordlist and picture memory), working memory tasks (N-back, symbol recall) and motor learning (maze task, pursuit rotor). Visuospatial abilities were assessed by tests with high visual scanning components (tangled lines and Symbol Digit Substitution Test). RESULTS: Episodic memory was impaired by biperiden. Rivastigmine impaired recognition parts of the episodic memory performance. Working memory was non-significantly impaired by biperiden and not affected by rivastigmine. Motor learning as well as visuospatial processes were impaired by biperiden and improved by rivastigmine. CONCLUSIONS: These results implicate acetylcholine as a modulator not only of memory but also of visuospatial abilities. PMID- 16041535 TI - Phenotypic studies on dopamine receptor subtype and associated signal transduction mutants: insights and challenges from 10 years at the psychopharmacology-molecular biology interface. AB - BACKGROUND: Mutants with targeted gene deletion ('knockout') or insertion (transgenic) of D1, D2, D3, D4 and D5 dopamine (DA) receptor subtypes are complemented by an increasing variety of double knockout and transgenic 'knockout' models, together with knockout of critical components of DA receptor signalling cascades such as G alpha(olf)[G gamma7], adenylyl cyclase type 5, PKA [RIIbeta] and DARPP-32. However, it is increasingly recognised that these molecular techniques have a number of inherent limitations. Furthermore, there are poorly understood methodological factors that contribute to inconsistent phenotypic findings between laboratories. OBJECTIVE: This review seeks to document the impact of DA receptor subtype and related transduction mutants on our understanding of the behavioural roles of these entities, primarily at the level of unconditioned psychomotor behaviour. METHODS: It includes ethologically based and orofacial movement studies in our own laboratories, since these are the only studies to systematically compare each of the D1, D2, D3, D4 and D5 receptor and DARPP-32 signal transduction 'knockouts'. DISCUSSION: There is a particular emphasis on identifying methodological factors that might influence phenotypic effects and account for inconsistencies. The findings are offered empirically to (1) specify the extent of phenotypic diversity among individual DA receptor subtypes and transduction components and (2) indicate relationships between D1, D2, D3, D4 and D5 receptor subtype proteins, associated G alpha(i)/G alpha(s)/G alpha(olf)[G gamma7]-adenylyl cyclase type 5-PKA [RIIbeta]-DARPP-32 signalling cascades and behaviour. The findings are also offered heuristically as a base for such phenotypic comparisons at additional levels of behaviour so that a yet more complete phenotypic profile might emerge. PMID- 16041536 TI - Marked differences in the IGF system that are associated with migration in comparable populations of Gujaratis living in Sandwell, UK, and Gujarat, India. AB - AIMS/HYPOTHESES: We previously reported independent links between the IGF system and the development of impaired glucose tolerance and cardiovascular risk. This study tests the hypothesis that the lifestyle change which accompanies population migration, with attendant increases in cardiovascular risk, is reflected by changes in the IGF system. MATERIALS AND METHODS: We compared a specific Gujarati community in Sandwell, UK (n=205), with people still resident in the same villages of origin near Navsari, India (n=246). We performed anthropometry and measured fasting plasma insulin, IGF-I, insulin-like growth factor binding protein (IGFBP)-1 and IGFBP-3. RESULTS: Daily calorie intake, BMI and WHR were significantly higher in UK Gujaratis than in Indian Gujaratis. IGFBP-1 was significantly lower in UK migrants (mean 29.5 [95% CI 25.9-33.0] vs 56.5 [50.6 62.5] microg/l; F=48.4, p<0.001). Conversely, fasting insulin, IGFBP-3 and IGF-I were all higher in UK Gujaratis (mean IGF-I 145.9 [138.1-153.6]ng/ml in UK Gujaratis and 100.9 [94.6-107.3] ng/ml in Navsari Gujaratis; F=76.6, p<0.001). These differences were still apparent when adjustment was made for BMI by location for IGF-I (F=57.4, p<0.001) and IGFBP-3 (F=5.7, p=0.02), but were no longer apparent for IGFBP-1 and insulin. At the population level, the decrease in IGFBP-1 for a given increase in insulin was significantly smaller in UK Gujaratis, suggesting greater hepatic insulin resistance in this group. CONCLUSIONS/INTERPRETATION: Environmental factors have profound effects on circulating IGF system components and on the relationship between IGFBP-1, IGF-I and related metabolic variables. This may have long-term implications for the development of worsening glucose tolerance and cardiovascular disease. PMID- 16041537 TI - Comparison of thoracoscopic drainage with open thoracotomy for treatment of paediatric parapneumonic empyema. AB - The purpose of this study was to compare the efficacy of thoracoscopic drainage with open drainage of empyema in children. A retrospective case history audit was done of children presenting to a single major paediatric centre who underwent thoracoscopy drainage and decortication or open decortication and drainage (thoracotomy) between January 2000 and September 2002. Time to resolution of infection, duration of intercostal catheter (ICC) drainage, postoperative morphine requirements, and length of hospital admission were compared as primary measures of outcome. Thirty-three patients, 17 male and 16 female, aged between 1 month and 21 years were included in the study. Median age at surgery was 2.6 years. The location of the empyema was right-sided in 17 patients and left-sided in 16 patients. The empyema was drained by thoracoscopy in 11 patients, and 22 patients underwent thoracotomy. Two patients had thoracoscopy converted to thoracotomy for late-stage disease requiring greater surgical access. There were no differences between treatment groups with respect to duration of ICC drainage (p=0.6), duration of fever (p=0.6), length of stay (p=0.9), or postoperative morphine use (p=0.2). However, overall pain scores were lower in the thoracoscopy group, particularly on days 2 and 3; this approached statistical significance (p=0.07). This study has demonstrated that thoracoscopic drainage is an effective procedure for treating empyema in children. It is less invasive than open thoracotomy and is associated with less patient discomfort and less severe pain as measured by objective pain scores. We advocate thoracoscopic drainage for the majority of patients with empyema, except for those with advanced disease. PMID- 16041538 TI - Primary pulmonary hydatid cysts in children-a report of three cases. AB - Three children with a total of six primary pulmonary hydatid cysts, all of whom underwent surgical management, are presented. Of these six hydatid cysts, two were treated with thoracotomy with cyst enucleation and capittonage, three with thoracoscopy-assisted minithoracotomy with enucleation and capittonage, and one with limited resection. PMID- 16041539 TI - Reimplantation of oesophageal bronchus following a type III oesophageal atresia repair. AB - Oesophageal atresia can be associated with communicating bronchopulmonary foregut malformations, the most common being the oesophageal bronchus. Lung resection may be mandatory, but conservative treatment correcting the abnormal airway has been proposed, raising the difficult issue of perioperative ventilatory support. The authors report a case of successful reimplantation of oesophageal bronchus revealed after surgical cure of an oesophageal atresia with tracheo-oesophageal fistula. PMID- 16041540 TI - Characterization of a C3-like cDNA in a coral: phylogenetic implications. AB - C3, C4, and C5 are thiolester-containing proteins (TEPs) of vertebrate complement. The identification of the molecular origin of the TEP family, and more specifically the ancestor protein of complement components C3, C4, and C5, remains a fundamental question. The prevailing paradigm suggests that duplication and divergence of these proteins occurred after the deuterostome split in phylogeny. It is believed that the ancestor of thiolester-containing complement proteins was alpha-2-macroglobulin (A2M)-like, a noncomplement-related protein. Here we describe a C3-like cDNA from a gorgonian coral, Swiftia exserta. This study provides evidence for the origins of a complement-related C3-like gene in the Precambrian period, predating both protostomes and deuterostomes. Furthermore, one may speculate that complement-like opsonic reactions were evolving at the earliest stages of metazoan evolution. This calls for a reassessment of the present concepts concerning the origins and evolution of TEPs. PMID- 16041541 TI - Interferon-alpha/beta upregulate IL-15 expression in vitro and in vivo: analysis in human hepatocellular carcinoma cell lines and in chronic hepatitis C patients during interferon-alpha/beta treatment. AB - Type I interferon (IFN) possesses antiviral and antitumor activities and also having an immune regulatory effect, activating cellular immune response and upregulating several cytokines. Recent study has shown that type I IFN upregurates the dendritic cell production of IL-15 capable of activating natural killer cells and CD8+ memory T lymphocytes. However, it is still unknown if type I IFN induces IL-15 production in non-immune cells and if type I IFN affects IL 15 production in vivo. The present study investigated the effect of type I IFNs on IL-15 expression in hepatocellular carcinoma (HCC) cell lines in vitro and in patients with chronic hepatitis C in vivo. When three HCC cell lines, Huh7, HepG2, and JHH4 were cultured in vitro, IFN upregulation of IL-15 expression was observed at both the mRNA and protein levels. In experiments using Huh7 cells, upregulation of IL-15 expression occurred within 24 h of the start of IFN stimulation, and both IFN-alpha and -beta dose-dependently increased IL-15 production in the range from 100 U/ml to 10,000 U/ml of concentration. IFN-beta showed stronger activity in IL-15 production induction in vitro than IFN-alpha. For in vivo examination, sera were obtained from 21 chronic hepatitis C patients treated with IFN and 29 healthy individuals, and the serum IL-15 level was quantified by ELISA. The serum IL-15 level of chronic hepatitis C patients before IFN treatment was similar to that of the healthy controls and significantly increased only during the IFN administration period. These results confirm that IFN-alpha/beta induce IL-15 production and also suggest that IL-15 may be associated with type I IFN-induced immune response. PMID- 16041543 TI - Tailgut cyst initially misdiagnosed as ovarian tumor. AB - INTRODUCTION: Tailgut cyst (retrorectal cystic hamartoma) is an uncommon congenital lesion that arises from remnants of the embryonic post-anal gut. It is usually benign and located in the retrorectal/presacral space. CASE REPORT: A 45 year-old woman presented with lower abdominal discomfort, urinary frequency and right-sided pelvic mass. Digital rectal examination and computerized tomography scan demonstrated a multicystic complex mass occupying the region of the right ovary and extending into the retrorectal/presacral space. The initial diagnosis was neoplasm of the right ovary. At laparotomy, the mass was lying free from apparently normal internal genital organs. Excision of the mass from the right pelvis and retrorectal/presacral space and total abdominal hysterectomy were performed. Microscopic examination revealed that the wall of the cystic mass consists of a lining epithelium composed of columnar and squamous epithelium and a stroma composed of fibrous tissue containing scattered discontinuous bundles of smooth muscle fibers. Based on these histological findings, the final diagnosis was tailgut cyst. CONCLUSION: Tailgut cyst is an uncommon entity that should be included in the differential diagnosis of retrorectal/presacral mass. The treatment of choice is complete surgical excision. PMID- 16041544 TI - Tracing Mississippi River influences in estuarine food webs of coastal Louisiana. AB - The Breton Sound estuary in southern Louisiana receives large amounts of Mississippi River water via a controlled diversion structure at the upstream end of the estuary. We used stable isotopes to trace spatial and seasonal responses of the downstream food web to winter and spring introductions of river water. Analysis of delta13C, delta15N, and delta34S in the common local consumers such as grass shrimp (Palaemonetes sp.), barnacles (Balanus sp.), and small plankton feeding fish (bay anchovies, Anchoa mitchilli) showed that the diversion was associated with two of the five major source regimes that were supporting food webs: a river regime near the diversion and a river-influenced productive marsh regime farther away from the diversion. Mixing models identified a third river influenced source regime at the marine end of the estuary where major natural discharge from the Bird's Foot Delta wraps around into estuarine waters. The remaining two source regimes represented typical estuarine conditions: local freshwater sources especially from precipitation and a brackish source regime representing higher salinity marine influences. Overall, the Mississippi River diversion accounted for 75% of food web support in the upper estuary and 25% in the middle estuary, with influence strongest along known flow pathways and closest to the diversion. Isotopes also traced seasonal changes in river contributions, and indicated increased plant community productivity along the major flow path of diversion water. In the Breton Sound estuary, bottom-up forcing of food webs is strongly linked to river introductions and discharge, occurring in spatial and temporal patterns predictable from known river input regimes and known hydrologic circulation patterns. PMID- 16041545 TI - Resolving temporal variation in vertebrate diets using naturally occurring stable isotopes. AB - Assessments of temporal variation in diets are important for our understanding of the ecology of many vertebrates. Ratios of naturally occurring stable isotopes in animal tissues are a combination of the source elements and tissue specific fractionation processes, and can thus reveal dietary information. We review three different approaches that have been used to resolve temporal diet variation through analysis of stable isotopes. The most straightforward approach is to compare samples from the same type of tissue that has been sampled over time. This approach is suited to address either long or short-term dietary variation, depending on sample regime and which tissue that is sampled. Second, one can compare tissues with different metabolic rates. Since the elements in a given tissue have been assimilating during time spans specific to its metabolic rate, tissues with different metabolic rates will reflect dietary records over different periods. Third, comparisons of sections from tissues with progressive growth, such as hair, feathers, claws and teeth, will reveal temporal variation since these tissues will retain isotopic values in a chronological order. These latter two approaches are mainly suited to address questions regarding intermediate and short-term dietary variation. Knowledge of tissue specific metabolic rates, which determine the molecular turnover for a specific tissue, is of central importance for all these comparisons. Estimates of isotopic fractionation between source and measured target are important if specific hypotheses regarding the source elements are addressed. Estimates of isotopic fractionation, or at least of differences in fractionation between tissues, are necessary if different tissues are compared. We urge for more laboratory experiments aimed at improving our understanding of differential assimilation of dietary components, isotopic fractionation and metabolic routing. We further encourage more studies on reptiles and amphibians, and generally more studies utilizing multiple tissues with different turnover rates. PMID- 16041546 TI - On the discriminability of hROIs, human visually selected regions-of-interest. AB - Many studies have tried to answer an important question: is it possible to predict human visually selected regions-of-interest (hROIs)? hROIs are defined as the loci of eye fixations and they can be analyzed by their spatial distribution over the visual stimulus and their temporal ordering. We used a simplified set of geometrical spatial kernels and linear filter models as bottom-up conspicuity operators that produce algorithmically selected regions-of-interest, aROIs. As a direct approach we measured the ability of these aROIs to predict human scanpaths. The level of prediction is measured by two similarity indices: Sp for spatial similarity and Ss for temporal ordering similarity. At the same time we assessed the discriminability of the hROI loci, in terms of conspicuity, with respect to non-selected (not of interest) regions of an image. We prove that this discrimination is possible and further correlates with the positional similarity index Sp. Other human scanpath experimental conditions are presented in parsing diagrams and discussed. A general top-down/bottom-up scanpath model is finally formulated. PMID- 16041547 TI - Cytochrome P450 mRNA expression in peripheral blood lymphocytes as a predictor of enzyme induction. AB - OBJECTIVE: Previous reports have supported the concept that messenger ribonucleic acid (mRNA) concentrations for cytochrome P450 (CYP) enzymes in peripheral blood mononuclear cells may be predictive of systemic enzyme activity. We investigated whether changes in mRNA expression for CYP1A2,CYP2C19, CYP2D6 and CYP3A4 in peripheral blood lymphocytes (PBLs) may serve as surrogate markers for changes in CYP enzyme activity following the administration of rifampin. METHODS: On day 1 and day 9 of the study, 12 healthy volunteers were administered caffeine 100 mg, debrisoquine 10 mg and omeprazole 40 mg orally, along with midazolam 0.025 mg/kg intravenously. Blood samples and urine were collected for 8 h after drug administration. The subjects took rifampin 300 mg (n = 6) or 600 mg (n = 6) daily on days 2-8. Total RNA was isolated from PBLs on day 1 and day 9, and mRNA expression for the CYP enzymes and hGAPDH were determined by means of quantitative, real-time, reverse-transcriptase polymerase chain reaction. CYP1A2 activity was estimated by calculating the plasma paraxanthine to caffeine AUC ratio (caffeine metabolic ratio; CMR), CYP2C19 activity by the 2-h omeprazole hydroxylation index (HI), CYP2D6 activity by the urinary debrisoquine recovery ratio (DBRR) and CYP3A4 activity by midazolam clearance. RESULTS: Median midazolam clearance (0.362 to 0.740 l/kg/h), omeprazole HI (0.752 to 0.214), CMR (0.365 to 0.450) and DBRR (0.406 to 0.479) all changed significantly following rifampin, consistent with the expected enzyme induction. CYP1A2,CYP2D6 and CYP3A4 mRNA content were measurable in all samples. CYP2C19 mRNA was inconsistently detectable. There were no significant correlations between changes in enzyme activity and mRNA expression by Spearman's rank order correlation. CONCLUSION: The results do not support the use of mRNA expression assays for CYP1A2, CYP2C19, CYP2D6 and CYP3A4 enzymes in PBLs as surrogates for quantifying changes in systemic enzyme activity in the setting of enzyme induction. PMID- 16041548 TI - Epidermoid cyst of the brain stem symptomatic in childhood. AB - BACKGROUND: Epidermoid cysts may remain asymptomatic for a long period of time due to their slowly growing pattern corresponding with the normal human skin turnover time and due to soft and light cyst content. They do not cause compression of neural and vascular structures initially that is why almost all of the cases in the literature are diagnosed during adulthood. METHODS: We report here an epidermoid cyst in childhood, which was located in the medulla oblongata, unusually and atypically with liquefied cyst content. The liquefaction may occur due to an intrauterine or early childhood infection. The reported case also suffered previously a severe respiratory infection. Although the cyst is located in and around a highly eloquent neural area, plasticity of the brain stem prevented neurological deterioration due to this very slow growing extraaxial mass lesion. The ordinary cyst content found in the center of the cyst cavity during the operation suggested that the same ordinary material, which was previously at the periphery, ran to get liquefied in time. CONCLUSION: We suggest that the symptoms of this present case appeared very early due to liquefaction of the cyst content with compression and displacement of the brain stem caudally. The recent infection process may predispose the pathological condition. PMID- 16041549 TI - [Quantitative evaluation of thermosensitivity in patients with mandibular fractures]. AB - AIM: The temperature and pain thresholds of 30 patients with mandibular fractures were evaluated with a specially developed measuring device in a prospective study to examine neurosensory disturbances of the inferior alveolar nerve (IAN). PATIENTS AND METHODS: Measurements in 30 patients with 45 mandibular fractures were done before, immediately after, and over the course of half a year after operation. Outcomes of the thermosensitivity measurement (TSM) were compared to the results of the two point discrimination (2-PD) test, sharp/blunt test (SBT), and questioning the patients about their subjective feelings (SF). RESULTS: Post traumatically, neurosensory testing revealed up to 2.8 degrees C (mean) increased thresholds as compared with the intact side. By contrast, post operative diagnostics clearly revealed up to 8.1 degrees C (mean) increased thresholds. Compared with the clinical sensory tests, the post-therapeutic (3-6 months postoperative) measurements with the TSM device proved a resensitization and threshold decrease by up to 3 degrees C. We found more exact and reproducible results of the TSM in comparison to those of the 2-PD and SBT for the evaluation of sensory disturbances of the IAN. Close agreements between the results of the TSM and the SF could be detected. CONCLUSION: Because of its sensitivity and accuracy in the quantitative detection of temperature and pain thresholds, the TSM is particularly suitable for evaluating neurosensory deficits and monitoring their progression. PMID- 16041550 TI - Subependymal giant cell astrocytoma and concordant expression in a disease of variable penetrance. PMID- 16041551 TI - Success of endoscopic third ventriculostomy in children less than 2 years of age. AB - Current literature reveals different opinions about the effectiveness of endoscopic third ventriculostomy in the treatment of hydrocephalus in children less than 2 years of age. Performing a retrospective evaluation of our own experience in this age group, we aimed to contribute to the growing data on the controversial issues related to this procedure in children. In a series of 97 endoscopic third ventriculostomy procedures, 25 were performed in children less than 2 years of age as an initial treatment for hydrocephalus. A retrospective analysis of our data revealed that the overall success rate of endoscopic third ventriculostomy in this age group was 56%. However, analysis of the results in subgroups with different etiologies of hydrocephalus showed that the success rate of the procedure was 83% in patients with defined anatomic obstruction, 66.6% in post-hemorrhagic hydrocephalus, 50% in infection related hydrocephalus and 41.6% in hydrocephalus accompanied by myelomeningocele. This article considers our data and the features of endoscopic third ventriculostomy procedure in this age group, with a detailed review of the literature. In our experience, the success of endoscopic third ventriculostomy is etiology related rather than age-dependent. We suggest that there are no grounds for denying children younger than 2 years this chance for a shunt-free life. PMID- 16041552 TI - Modified radical lymphadenectomy (D1.5) for T2-3 gastric cancer. AB - BACKGROUND: The operative mortality in gastric cancer surgery has been reported to be higher with D2 lymphadenectomy than with D1 in the West. The modified radical lymphadenectomy (D1.5) may be safer than D2 under these circumstances. This study was aimed to determine whether D1.5 would deteriorate long-term survival as compared with D2. METHOD: Since the concept of the extent of lymphadenectomy varied among the surgeons, 461 patients who underwent curative gastrectomy for T2-4 gastric adenocarcinoma were retrospectively categorized into three groups according to the surgeon: D1 with dissection along the left gastric and common hepatic arteries (D1.5); lymphadenectomy between D1.5 and D2; D2 or more extended dissection. RESULTS: No differences were found in the survival rates among the three groups within each of the T2a, T2b, and T3 categories. According to a multivariate analysis using Cox's proportional hazard model, the classification according to the surgeons had no survival impact (p>0.8). CONCLUSION: D1.5 lymphadenectomy resulted in a survival rate that was almost equal to that of D2. The use of D1.5 instead of D2 can be an attractive option to be compared with D1 in future trials. PMID- 16041553 TI - Risk factors influencing the early outcome results after laparoscopic repair of perforated duodenal ulcer and their predictive value. AB - BACKGROUND AND AIMS: Clear patient selection criteria and indications for laparoscopic repair of perforated duodenal ulcers are necessary. The aims of our study are to report the early outcome results after operation and to define the predictive values of risk factors influencing conversion rate and genesis of suture leakage. PATIENTS/METHODS: Sixty nonrandomly selected patients operated on laparoscopically in a tertiary care academic center between October 1996 and May 2004 for perforated duodenal ulcers were retrospectively analyzed. The primary outcome measures included the duration of symptoms, shock, underlying medical illness, ulcer size, age, Boey score, and the collective predictive value of these variables for conversion and suture leakage rates. RESULTS: Laparoscopic repair was completed in 46 patients (76.7%). Fourteen patients (23.3%) underwent conversion to open repair. Eight patients (13.3%) had postoperative complications. Suture leakage was confirmed in four patients (6.7%). Hospital stay was 7.8+/-5.3 days. There was no mortality. Patients with an ulcer perforation size of >8 mm had a significantly increased risk for conversion to open repair (p<0.05): positive predictive value (PPV) 75%, sensitivity 27%, specificity 98%, and negative predictive value (NPV) 85%. The significance of ulcer perforation size was confirmed by a stepwise logistic regression test (p=0.0201). All patients who developed suture leakage had acute symptoms for >9 h preoperatively (p<0.001): PPV 31%, specificity 84%, sensitivity 100%, and NPV 100%. Conversions happened with surgeons whose previous experience involved 1.8+/ 2.3 cases compared to 3.9+/-2.9 cases in successful laparoscopic repair (p=0.039, t test). CONCLUSIONS: Ulcer perforation size of >8 mm is a significant risk factor influencing the conversion rate. An increase in the suture leakage rate is predicted by delayed presentation of >9 h. PMID- 16041554 TI - Distribution of the vanilloid (capsaicin) receptor type 1 in the human stomach. AB - Vanilloid receptor type 1 (TRPV1) is expressed in a capsaicin-sensitive and peptide-containing sub-population of primary sensory nerves that in the rat stomach seems involved in regulation of chlorhydropeptic secretion and gastroprotection. Our aim was to identify which cell types express TRPV1 in the human stomach in order to gain a better insight in the role of this receptor in the regulation of HCl secretion. Immunohistochemistry, by using three different commercially available anti-capsaicin antibodies, in situ hybridisation and Western blot analysis were performed on fragments surgically obtained from the gastric body on the large curvature. TRPV1 labelling was found in the parietal cells at the level of intra-cytoplasmatic granules matching mitochondrial features and distribution. Immunolabelled neurons and nerve fibres were also seen, the latter numerous in the submucosa and mucosa and often ending close to the parietal cells. TRPV1 presence was confirmed by Western blot analysis and in situ hybridisation. TRPV1 presence in nerve structures and parietal cells suggests the possibility of a combined effect of both neuronal and epithelial TRPV1 on chlorhydropeptic secretion. The presumed TRPV1 mitochondrial location inside parietal cells is in favour of the existence of a local pathway of auto regulation of HCl secretion. Therefore, TRPV1 might modulate chlorhydropeptic secretion in the human stomach through more complex pathways than previously thought. PMID- 16041555 TI - Aneurysm of an aberrant right subclavian artery successfully excluded by a thoracic aortic stent graft with supra-aortic bypass of three arch vessels. AB - An aberrant right subclavian artery (ARSA) arising from a left-sided aortic arch is the fourth most common aortic arch anomaly. Aneurysmal dilatation of the ARSA requires treatment because of the associated risk of rupture. We present a case where supra-aortic bypass of the arch vessels was performed to facilitate exclusion of the aneurysm by a thoracic aortic stent graft. PMID- 16041556 TI - Treatment of neuroendocrine cancer metastatic to the liver: the role of ablative techniques. AB - Carcinoid tumors and islet cell neoplasms are neuroendocrine neoplasms with indolent patterns of growth and association with bizarre hormone syndromes. These tumors behave in a relatively protracted and predictable manner, which allows for multiple therapeutic options. Even in the presence of hepatic metastases, the standard of treatment for neuroendocrine malignancy is surgery, either with curative intent or for tumor cytoreduction, i.e., resection of 90% or more of the tumor volume. Image-guided ablation, as either an adjunct to surgery or a primary treatment modality, can be used to treat neuroendocrine cancer metastatic to the liver. Image-guided ablative techniques, including radiofrequency ablation, alcohol injection, and cryoablation, can be used in selected patients to debulk hepatic tumors and improve patient symptoms. Although long-term follow-up data are not available, the surgical literature indicates that significant ablative debulking may improve patient survival. In this review, we discuss metastatic neuroendocrine disease and its treatment options, especially image-guided ablative techniques. PMID- 16041557 TI - A check on the memory deficit hypothesis of obsessive-compulsive checking. AB - A number of recent studies have challenged the hypothesis that patients with obsessive-compulsive disorder (OCD) display global memory deficits. An alleviated form of the memory deficit hypothesis posits that OCD patients share deficits to vividly recall memory episodes. According to the latter view, checking rituals can be understood as counter-productive coping strategies to "enrich" memory episodes in order to make them more distinctive. A source memory task was administered to 27 OCD (17 checkers) and 51 healthy participants. Along with confidence judgments, a remember-know procedure was employed to assess whether OCD patients display problems with conscious/vivid recollection. Patients with or without checking compulsions did not exhibit differences to controls on source memory accuracy and meta-memory. Patients forgot more self-generated items, which, however, was related to comorbid depressive but not OCD symptoms. Findings challenge the ubiquity of memory deficits in OCD. To account for the inconclusive pattern of results in the literature, it is suggested that patients mistrust their memories and adopt checking rituals only when perceived responsibility is inflated. PMID- 16041558 TI - Association of personality disorders with Type A and Type B alcoholics. AB - Personality disorders frequently occur as comorbid disorder in alcohol-dependent subjects. Antisocial personality was described as an important characteristic in Cloninger's Type 2 and Babor's Type B subjects. The impact of other personality disorders on these alcoholism typologies, their pathogenesis and prognosis is, however, still unclear. The present study investigated the prevalence of personality disorders in 237 (194 males) detoxified alcohol-dependent patients after subtyping this sample according to Babor's Type A/B following the criteria suggested by Schuckit et al. (1995). Personality disorders were assessed with the SCID-II (DSM-IV). In all, 160 patients (68 %) could be classified as Type A, and 77 (32 %) as Type B. Type B subjects were younger, had an earlier onset,more alcohol intake and a more severe course of alcohol dependence. Type B patients had significantly more often any cluster A and B personality disorder, and significantly specifically more often a borderline, antisocial and avoidant personality disorder. There were no statistical differences concerning the other personality disorders. In summary, the Type A/B dichotomy using the criteria of Schuckit et al. (1995) was replicated successfully. Differences concerning cluster B personality disorder prevalence of Type B subjects demonstrated that these subjects are significantly more often affected from borderline and antisocial personality disorder. The impact of other personality disorders does not play a substantial role in subtyping alcoholics. PMID- 16041559 TI - Atypical depressive syndromes in varying definitions. AB - BACKGROUND: Atypical depression (AD) exhibits distinct patterns of gender,bipolar II disorder, genetic, and neuro-biological measures. Using prospective data from a community sample, this paper identifies criteria (and correlates) for an AD syndrome that maximizes the association with female sex and bipolar-II. METHODS: The Zurich cohort study is composed of 591 subjects selected from a population based cohort of young adults in the canton of Zurich in Switzerland, screened in 1978 and followed with six interviews through 1999. Seven definitions of atypical depression were tested, using varying combinations of vegetative symptoms and mood reactivity. RESULTS: The atypical definitions using 2 of 3 (fatigue, overeating, oversleeping) or 2 of 2 (overeating, oversleeping) vegetative symptoms showed the strongest association with gender, bipolarity, and family history of mania. The 2/3 definition was chosen for further analysis due to its high sensitivity for identifying these characteristics. This syndrome had cumulated weighted prevalence of 16.4% (males 9.7%, females 23%); when associated with major depressive episodes, 8.2% (males 3.2%, females 15.1%). AD patients were characterized by high treatment rates, severity, and work impairment, early age of onset and long illness. AD was comorbid with social phobia, binge eating, neurasthenia, migraine headache, and subjective cognitive impairment. PMID- 16041560 TI - Anthropometric characteristics of the public arch and proper function of the defense mechanisms against hernia formation. PMID- 16041561 TI - Porcelain heart: a case of massive myocardial calcification. AB - Reports of massive myocardial calcification are limited mainly to case reports and this type intracardiac calcifications have been usually identified postmortem. We present a very interesting case of massive calcification of the left ventricular myocardium, interventricular and interatrial septae causing restrictive physiology and coronary artery obstruction in a 46-year-old Turkish woman. The diagnosis of myocardial calcification was suspected from chest X-ray and confirmed with computerized tomography and cardiac catheterization. The findings of the chest computed tomography were very similar to those previously reported cases of endomyocardial fibrosis (EMF) with massive calcification of the left ventricle. PMID- 16041562 TI - MRI demonstration of acute myocardial infarction due to posttraumatic coronary artery dissection. AB - A case of left ventricular lateral wall myocardial infarction in the distribution of circumflex coronary artery (LCX) was demonstrated by magnetic resonance imaging in a 55-year-old woman. Dissection of the proximal LCX due to blunt chest trauma was followed by percutaneous coronary artery stenting. MR (magnetic resonance) imaging of myocardial infarction is reviewed. PMID- 16041563 TI - Detection of small vessels with electron beam computed tomographic angiography using 1.5 and 3 mm collimator protocols. AB - OBJECTIVES: To evaluate the effect of scanner collimation on the ability to detect small cardiac vessels using electron beam CT coronary angiography (EBA). MATERIALS AND METHODS: EBA scans from 40 patients who underwent study on two separate occasions with 3 mm (initial scan) and 1.5 mm (follow-up scan) collimation protocols were analyzed. Vessels of <2 mm in diameter were identified. RESULTS: The 1.5 mm collimation allowed 3-D visualization of 129 vessels<2 mm in diameter, while 3 mm collimation only allowed visualization of 89 vessels (p<0.001). The right coronary artery branches and distal LAD segments though were not displayed satisfactorily in almost half of the 3-D studies with either protocol. CONCLUSIONS: There was significant improvement in detection of small cardiac vessels with a 1.5 mm collimation EBA protocol compared to a 3 mm protocol. Both protocols though were insufficient for reliable visualization of the right coronary artery branches and distal LAD segments. PMID- 16041564 TI - Receptor-mediated transport and deposition of complement component C3 into developing chicken oocytes. AB - Immunological resistance of the chick embryo is dependent upon IgG present in the yolk of the layed egg. Here we show that complement factor 3 (C3), a key component of the humoral complement system, is a yolk component of chicken eggs. C3 is transported into oocytes by LR8-mediated endocytosis. LR8 also binds and transports other major yolk components such as vitellogenin, very-low-density lipoprotein, and alpha(2)-macroglobulin. Expression studies of LR8 during chicken development and oocyte maturation, in combination with studies on the uptake of individual yolk components, suggest the following model for oocyte maturation in the chicken: all oocytes present in the ovary contain high levels of LR8 mRNA and protein long before the onset of oocyte maturation. Selected oocytes gain access to yolk precursors, and LR8 binds, internalizes, and deposits the major yolk components in the ratio of their relative abundance in the accessible pool. PMID- 16041565 TI - Bacterial signal peptide recognizes HeLa cell mitochondrial import receptors and functions as a mitochondrial leader sequence. AB - Phage display was used to identify new components of the mammalian mitochondrial receptor complex using Tom20 as a binding partner. Two peptides were identified. One had partial identity (SMLTVMA) with a bacterial signal peptide from Toho-1, a periplasmic protein. The other had partial identity with a mitochondrial inner membrane glutamate carrier. The bacterial signal peptide could carry a protein into mitochondria both in vivo and in vitro. The first six residues of the sequence, SMLTVM, were necessary for import but the two adjacent arginine residues in the 30-amino-acid leader were not critical for import. The signal peptides of Escherichia coli beta-lactamase and Bacillsus subtilis lipase could not carry proteins into mitochondria. Presumably, the Toho-1 leader can adopt a structure compatible for recognition by the import apparatus. PMID- 16041566 TI - Circulating serotonin in vertebrates. AB - The role of circulating serotonin is unclear and whether or not serotonin is present in the blood of non-mammalian species is not known. This study provides the first evidence for the presence of serotonin in thrombocytes of birds and three reptilian species, the endothermic leatherback sea turtle, the green sea turtle and the partially endothermic American alligator. Thrombocytes from a fresh water turtle, American bullfrog, Yellowfin tuna, and Chinook salmon did not contain serotonin. Serotonin is a vasoactive substance that regulates skin blood flow, a major mechanism for endothermic body temperature regulation, which could explain why circulating serotonin is present in warm-blooded species. The temperature sensitivity of human blood platelets with concomitant changes in serotonin content further supports a link between circulating serotonin and thermoregulation. Phylogenetic comparison of the presence of circulating serotonin indicated an evolutionary divergence within reptilian species that might coincide with the emergence of endothermy. PMID- 16041567 TI - Nucleotide analogues as probes for DNA polymerases. AB - Transmission of the genetic information from the parental DNA strand to the offspring is crucial for the survival of any living species. In nature, all DNA synthesis in DNA replication, recombination and repair is catalyzed by DNA polymerases and depends on their ability to select the canonical nucleobase pair from a pool of structurally similar building blocks. Recently, a wealth of valuable new insights into DNA polymerase mechanisms have been gained through application of carefully designed synthetic nucleotides and oligonucleotides in functional enzyme studies. The applied analogues exhibit features that differ in certain aspects from their natural counterparts and, thus, allow investigation of the involvement and efficacy of a chosen particular aspect on the entire complex enzyme mechanism. This review will focus on a depiction of the efforts that have been undertaken towards the development of nucleotide analogues with carefully altered properties. The different approaches will be discussed in the context of the motivation and the problem under investigation. PMID- 16041568 TI - The silence of the ribosomal RNA genes. AB - Over the past decade emerging evidence has indicated that epigenetic factors control and regulate nuclear processes. The genes encoding ribosomal RNA (rRNA) represent an ideal model to study how epigenetics and chromatin can modulate gene expression. The reason for this is that in each cell, the rRNA genes exist in two distinct types of chromatin structure: an 'open' one corresponding to transcriptionally active genes and a 'closed' one representing the silent genes. Recent studies indicate that an epigenetic network mediates the transcriptional state of rDNA. Interplay of DNA methylation, histone modification and chromatin remodeling activities establishes silencing at the rDNA locus in higher eukaryotes as well as at the underdominant genes in hybrid cells. The aim of this review is to summarize current knowledge about the active and silent states of rRNA genes and of nucleolar organizing regions and to analyze the mechanisms involved in the establishment and inheritance of rDNA silencing. PMID- 16041570 TI - Efficient induction of ginsenoside biosynthesis and alteration of ginsenoside heterogeneity in cell cultures of Panax notoginseng by using chemically synthesized 2-hydroxyethyl jasmonate. AB - Chemically synthesized 2-hydroxyethyl jasmonate (HEJA) was for the first time employed to induce the ginsenoside biosynthesis and to manipulate the product heterogeneity in plant cell cultures. The dose response and timing of HEJA elicitation were investigated in cell suspension cultures of Panax notoginseng. The optimal concentration and timing of HEJA addition for both cell growth and ginsenoside accumulation was identified to be 200 microM added on day 4. It was interestingly found that HEJA could stimulate ginsenosides biosynthesis and change their heterogeneity more efficiently than methyl jasmonate (MJA), i.e., the total ginsenoside content and the Rb/Rg ratio increased about 60 and 30% with HEJA elicitation than that by MJA, respectively. The activity of Rb1 biosynthetic enzyme, i.e., UDPG-ginsenoside Rd glucosyltransferase (UGRdGT), was also higher in the former case. A maximal production titer of ginsenoside Rg1, Re, Rb1, and Rd was 47.4+/-4.8, 52.3+/-4.4, 190+/-18, and 12.1+/-2.5 mg/l with HEJA elicitation, which was about 1.3-, 1.3-, 1.7-, and 2.1-fold than that using MJA, respectively. Early signal events in plant defense response, including oxidative burst and jasmonic acid (JA) biosynthesis, were also examined. Levels of H2O2 and NO in medium and L-phenylalanine ammonia lyase activity in cells were not affected by addition of MJA and HEJA. On the other hand, the JA content in cells was increased with external jasmonates elicitation, and it was inhibited with the addition of JA biosynthesis inhibitors. The results suggest that oxidative burst might not be involved in the jasmonates-elicited signal transduction pathway, and MJA and HEJA may induce the ginsenoside biosynthesis via induction of endogenous JA biosynthesis and key enzymes (such as UGRdGT) in the ginsenoside biosynthetic pathway of P. notoginseng cells. The information is useful for hyperproduction of plant-specific heterogeneous products. PMID- 16041571 TI - Global physiological understanding and metabolic engineering of microorganisms based on omics studies. AB - Through metabolic engineering, scientists seek to modify the metabolic pathways of living organisms to facilitate optimized, efficient production of target biomolecules. During the past decade, we have seen notable improvements in biotechnology, many of which have been based on metabolically engineered microorganisms. Recent developments in the fields of functional genomics, transcriptomics, proteomics, and metabolomics have changed metabolic engineering strategies from the local pathway level to the whole system level. This article focuses on recent advances in the field of metabolic engineering, which have been powered by the combined approaches of the various "omics" that allow us to understand the microbial metabolism at a global scale and to develop more effectively redesigned metabolic pathways for the enhanced production of target bioproducts. PMID- 16041572 TI - Binding of S-layer homology modules from Clostridium thermocellum SdbA to peptidoglycans. AB - S-layer homology (SLH) module polypeptides were derived from Clostridium josui xylanase Xyn10A, Clostridium stercorarium xylanase Xyn10B, and Clostridium thermocellum scafoldin dockerin binding protein SdbA as rXyn10A-SLH, rXyn10B-SLH, and rSdbA-SLH, respectively. Their binding specificities were investigated using various cell wall preparations. rXyn10A-SLH and rXyn10B-SLH bound to native peptidoglycan-containing sacculi consisting of peptidoglycan and secondary cell wall polymers (SCWP) prepared from these bacteria but not to hydrofluoric acid extracted peptidoglycan-containing sacculi (HF-EPCS) lacking SCWP, suggesting that SCWP are responsible for binding with SLH modules. In contrast, rSdbA-SLH interacted with HF-EPCS, suggesting that this polypeptide had an affinity for peptidoglycans but not for SCWP. The affinity of rSdbA-SLH for peptidoglycans was confirmed by a binding assay using a peptidoglycan fraction prepared from Escherichia coli cells. The SLH modules of SdbA must be useful for cell surface engineering in bacteria that do not contain SCWP. PMID- 16041573 TI - Methane production and microbial community structure in single-stage batch and sequential batch systems anaerobically co-digesting food waste and biosolids. AB - Anaerobic co-digestion of food waste and biosolids was carried out in sequential batch and single-stage batch systems in four treatments. Methane yield, which was used as a functional process parameter, differed between treatments, with the single-stage batch system generating lower volumes than the sequential batch systems. Volatile fatty acid (VFA) concentrations and pH in the leachate also differed between treatments. VFA concentrations were highest and methane generation yields lowest in the single-stage batch system in comparison to the sequential batch systems. The anaerobic microbial community structure of the domains Archaea and Bacteria, determined by denaturing gradient gel electrophoresis, differed between treatments and was correlated to a number of environmental parameters such as pH, VFA concentration and methane generation rate. Methane generation rate was significantly correlated to the community structure of Bacteria but not Archaea. This indicated that the substrates that are produced by acetogens (Bacteria) are important for the growth and community structure of the methanogens (Archaea). Community structure of Archaea changed over time, but this had no observable effect on functional ability based on methane yields. Microbial diversity (H') was shown to be not important in developing a functionally successful anaerobic microbial community. PMID- 16041574 TI - Recovery of pyruvic acid from biotransformation solutions. AB - The aim of this investigation was to separate pyruvic acid of biotransformation solutions from lactic acid through complex extraction. For this purpose, complex extraction was investigated from model solutions. Tri-n-octanylamine (TOA) was used as the extractant. The effects of various diluents, the stoichiometry of pyruvic acid to TOA, and the initial pH of the aqueous phase on the extraction process were investigated in this study. The effects of sodium hydroxide (NaOH) and trimethylamine (TMA) on the back extraction process were also studied, respectively. The optimal conditions attained from the model solutions proved efficient on the biotransformation solutions of different concentrations. A total recovery of 71-82% of pyruvic acid was obtained, whereas 89-92% of lactic acid was removed. The purity of pyruvic acid reached 97% after the removal of TMA by a simple distillation. PMID- 16041575 TI - Isolation of a bacterium that degrades urethane compounds and characterization of its urethane hydrolase. AB - A bacterium which degrades urethane compounds was isolated and identified as Rhodococcus equi strain TB-60. Strain TB-60 degraded toluene-2,4-dicarbamic acid dibutyl ester (TDCB) and accumulated toluene diamine as the degradation product. The enzyme which cleaves urethane bond in TDCB was strongly induced by acetanilide. The purified enzyme (urethane hydrolase) was found to be homogeneous on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The molecular weight was estimated to be 55 kDa. The optimal temperature and pH were 45 degrees C and 5.5, respectively. The enzyme hydrolyzed aliphatic urethane compound as well as aromatic ones. The activity was inhibited by HgCl(2), p chrolomercuribenzoic acid, and phenylmethylsulfonyl fluoride, suggesting that cysteine and/or serine residues play an important role in the activity. The enzyme catalyzed the hydrolysis of anilides, amides, and esters as well as TDCB. It was characterized as a novel amidase/esterase, differing in some properties from other known amidases/esterases. PMID- 16041576 TI - Rapid identification of target genes for 3-methyl-1-butanol production in Saccharomyces cerevisiae. AB - Extracellular conditions determine the taste of fermented foods by affecting metabolite formation by the micro-organisms involved. To identify targets for improvement of metabolite formation in food fermentation processes, automated high-throughput screening and cDNA microarray approaches were applied. Saccharomyces cerevisiae was cultivated in 96-well microtiter plates, and the effects of salt concentration and pH on the growth and synthesis of the fusel alcohol-flavoured substance, 3-methyl-1-butanol, was evaluated. Optimal fermentation conditions for 3-methyl-1-butanol concentration were found at pH 3.0 and 0% NaCl. To identify genes encoding enzymes with major influence on product formation, a genome-wide gene expression analysis was carried out with S. cerevisiae cells grown at pH 3.0 (optimal for 3-methyl-1-butanol formation) and pH 5.0 (yeast cultivated under standard conditions). A subset of 747 genes was significantly induced or repressed when the pH was changed from pH 5.0 to 3.0. Expression of seven genes related to the 3-methyl-1-butanol pathway, i.e. LAT1, PDX1, THI3, ALD4, ILV3, ILV5 and LEU4, strongly changed in response to this switch in pH of the growth medium. In addition, genes involved in NAD metabolism, i.e. BNA2, BNA3, BNA4 and BNA6, or those involved in the TCA cycle and glutamate metabolism, i.e. MEU1, CIT1, CIT2, KDG1 and KDG2, displayed significant changes in expression. The results indicate that this is a rapid and valuable approach for identification of interesting target genes for improvement of yeast strains used in industrial processes. PMID- 16041577 TI - Biotechnological production and applications of phytases. AB - Phytases decompose phytate, which is the primary storage form of phosphate in plants. More than 10 years ago, the first commercial phytase product became available on the market. It offered to help farmers reduce phosphorus excretion of monogastric animals by replacing inorganic phosphates by microbial phytase in the animal diet. Phytase application can reduce phosphorus excretion by up to 50%, a feat that would contribute significantly toward environmental protection. Furthermore, phytase supplementation leads to improved availability of minerals and trace elements. In addition to its major application in animal nutrition, phytase is also used for processing of human food. Research in this field focuses on better mineral absorption and technical improvement of food processing. All commercial phytase preparations contain microbial enzymes produced by fermentation. A wide variety of phytases were discovered and characterized in the last 10 years. Initial steps to produce phytase in transgenic plants were also undertaken. A crucial role for its commercial success relates to the formulation of the enzyme solution delivered from fermentation. For liquid enzyme products, a long shelf life is achieved by the addition of stabilizing agents. More comfortable for many customers is the use of dry enzyme preparations. Different formulation technologies are used to produce enzyme powders that retain enzyme activity, are stable in application, resistant against high temperatures, dust free, and easy to handle. PMID- 16041578 TI - Functional genomics of an anaerobic aromatic-degrading denitrifying bacterium, strain EbN1. AB - Nitrate-reducing bacteria of the recently recognized Azoarcus/Thauera group within the Betaproteobacteria contribute significantly to the biodegradation of aromatic and other refractory compounds in anoxic waters and soils. Strain EbN1 belongs to a distinct cluster (new genus) and is the first member of this phylogenetic group, the genome of which has been determined (4.7 Mb; one chromosome, two plasmids) by [Rabus R, Kube M, Heider J, Beck A, Heitmann K, Widdel F, Reinhardt R (2005) The genome sequence of an anaerobic aromatic degrading denitrifying bacterium, strain EbN1. Arch Microbiol 183:27-36]. Ten anaerobic and four aerobic aromatic-degradation pathways were recognized on the chromosome, with the coding genes mostly forming clusters. Presence of paralogous gene clusters (e.g. for anaerobic ethylbenzene degradation) suggests an even broader degradation spectrum than previously known. Metabolic versatility is also reflected by the presence of multiple respiratory complexes and is apparently controlled by an extensive regulatory network. Strain EbN1 is unique for its capacity to degrade toluene and ethylbenzene anaerobically via completely different pathways. Bioinformatical analysis of their genetic blueprints and global expression analysis (DNA-microarray and proteomics) of substrate-adapted cells [Kuhner S, Wohlbrand L, Fritz I, Wruck W, Hultschig C, Hufnagel P, Kube M, Reinhardt R, Rabus R (2005) Substrate-dependent regulation of anaerobic degradation pathways for toluene and ethylbenzene in a denitrifying bacterium, strain EbN1. J Bacteriol 187:1493-1503] indicated coordinated vs sequential modes of regulation for the toluene and ethylbenzene pathways, respectively. PMID- 16041579 TI - Complication rates of diagnostic cerebral arteriography in children. AB - BACKGROUND: Cerebral arteriography (CA) remains the gold standard in delineating both intra- and extracranial vascular anatomy. Most data relating to the safety of CA are drawn from studies of adult patients in whom the practicalities of the procedure, range of potential pathologies and comorbid factors are different from those in children. OBJECTIVE: To evaluate the current local and neurological complication rates of paediatric CA in the setting of a tertiary level children's hospital in the UK. MATERIALS AND METHODS: Data from patients who had undergone CA between January 1998 and July 2003 were reviewed. The medical, anaesthetic and nursing records, and angiography reports were reviewed for all identified patients. The following parameters were extracted and entered into a proforma: gender, age, ethnicity, diagnosis, cerebrovascular diagnosis, referral source, date of CA, number of vessels catheterized and local and neurological complications. RESULTS: A total of 176 CA studies were undertaken in 150 patients (median age 7.3 years, range neonate to 19 years; 83 males, 67 females) during the 5.5-year study period. The majority of referrals originated from the neurology (58%) and neurosurgery services (27.8%). No neurological complications or deaths occurred. Local complications occurred in eight children (4.5%). Five children had a groin haematoma and two had bleeding at the puncture site. A single child had a reduced pedal pulse distal to the site of catheterization, but Doppler imaging was normal. CONCLUSIONS: CA has a continuing role in the evaluation of cerebrovascular pathologies in children. Neurological complications are rare and local complications are not uncommon (around 5%), but are not usually serious. PMID- 16041580 TI - Multidetector CT angiography of pediatric vascular malformations and hemangiomas: utility of 3-D reformatting in differential diagnosis. AB - BACKGROUND: Vascular malformations can be difficult to diagnose and classify. Accurate classification is important because treatments and prognosis vary based on the type of lesion. Diagnosis is based on a combination of clinical features with a variety of imaging techniques, including US, MRI/MRA, CT, and conventional angiography. OBJECTIVE: We hypothesized that imaging features seen on 3-D reformatted images obtained with multidetector CT angiography (CTA) would aid in differential diagnosis of types of vascular anomalies. MATERIALS AND METHODS: We retrospectively reviewed CT scans of 11 patients with vascular lesions and pathologically proven diagnoses in which 3-D reformatting was obtained. RESULTS: The 3-D images accurately diagnosed hemangiomas and lymphangiomas in all cases, in contrast to diagnosis by clinical criteria and planar CT, which was difficult or inaccurate. The 3-D CTA did not aid in the distinction between venous malformations and arteriovenous malformations (AVM), which appeared similar. CONCLUSIONS: Our preliminary observations suggest that volume-rendered reformatting is helpful in categorizing clinically significant vascular head and neck lesions, resulting in more diagnostic value than planar CT imaging alone. In particular, 3-D CTA might allow accurate differentiation of hemangiomas from AVM, and of lymphangiomas from other types of lesions, which was, in our series, not possible using clinical examination or conventional planar CT angiography. PMID- 16041581 TI - Comparison of quiet breathing and controlled ventilation in the high-resolution CT assessment of airway disease in infants with cystic fibrosis. AB - BACKGROUND: Respiratory motion and low lung volumes limit the quality of HRCT examinations in infants and young children. OBJECTIVE: To assess the effects of respiratory motion and lung inflation on the ability to diagnose airway abnormalities and air trapping (AT) using HRCT in infants with cystic fibrosis (CF). MATERIALS AND METHODS: HRCT images of the lungs were obtained at four anatomical levels in 16 sedated children (age 2.4+/-1.1 years, mean+/-SD) with CF using controlled ventilation at full lung inflation (CVCT-I), at resting end exhalation (CVCT-E), and during quiet breathing (CT-B). Two blinded reviewers independently and then by consensus scored all images for the presence or absence of bronchiectasis (BE), bronchial wall thickening (BWT), and AT. RESULTS: Of the 64 images evaluated, BE was identified in 19 (30%) of the CVCT-I images compared to 6 (9%) of the CVCT-E images (P=0.006) and 4 (6%) of the CT-B images (P=0.044). AT was seen in 29 (45%) of the CVCT-E images compared to 14 (22%) of the CVCT-I images (P=0.012) and 12 (19%) of the CT-B images (P=0.012). There were no significant differences in the detection of BWT among the three methods. SUMMARY: In infants with CF, fully inflating the lung improved the ability to diagnose early BE, and obtaining motion-free images at end exhalation enhanced the detection of AT. PMID- 16041582 TI - Vesiculated alpha-tocopheryl succinate enhances the anti-tumor effect of dendritic cell vaccines. AB - Alpha tocopheryl succinate (alpha-TOS) is a non-toxic vitamin E analog under study for its anti-cancer properties. In an earlier study, we showed that alpha TOS, when used in combination with non-matured dendritic cells (nmDC) to treat pre-established tumors, acts as an effective adjuvant. In this study, we have used vesiculated alpha-TOS (Valpha-TOS), a more soluble form of alpha-TOS that is relevant for clinical use, in combination with dendritic cells to treat pre established murine tumors. We demonstrate that Valpha-TOS kills tumor cells in vitro and inhibits the growth of pre-established murine lung carcinoma (3LLD122) as effectively as alpha-TOS. The combination of Valpha-TOS plus non-matured or TNF-alpha-matured DC is more effective at inhibiting the growth of established tumors than Valpha-TOS alone. We also observed that Valpha-TOS induces expression of heat shock proteins in tumor cells and that co-incubation of non-matured DC with lysate derived from Valpha-TOS-treated tumor cells leads to DC maturation evidenced by up-regulation of co-stimulatory molecules and secretion of IL-12p70. This study therefore demonstrates the immunomodulatory properties of Valpha-TOS that may account for its adjuvant effect when combined with DC vaccines to treat established tumors. PMID- 16041583 TI - Reciprocal translocations: a trap for cytogenetists? AB - We report four cases of subjects with phenotypic abnormalities and mental retardation associated with apparently balanced translocations, two inherited and two de novo, which showed, by molecular analysis, a hidden complexity. All the cases have been analyzed with different molecular techniques, including array CGH, and in two of them the translocation breakpoints have been defined at the level of base pairs via studies in somatic hybrids containing single derivative chromosomes. We demonstrated that all the translocations were in fact complex rearrangements and that an imbalance was present in three of them, thus accounting for the phenotypic abnormalities. In one case, a Prader-Willi subject, we were not able to determine the molecular cause of his phenotype. This study, while confirming previous data showing unexpected complexity in translocations, further underscores the need for molecular investigations before taking for granted an apparently simple cytogenetic interpretation. PMID- 16041584 TI - Heightened stress response in primary fibroblasts expressing mutant eIF2B genes from CACH/VWM leukodystrophy patients. AB - Childhood ataxia with central nervous system hypomyelination (CACH), also called vanishing white matter (VWM) leukoencephalopathy, is a fatal genetic disease caused by mutations in eukaryotic initiation factor 2B (eIF2B) genes. The five subunits eIF2B factor is critical for translation initiation under normal conditions and regulates protein synthesis in response to cellular stresses. Primary fibroblasts from CACH/VWM patients and normal individuals were used to measure basal eIF2B activity as well as global protein synthesis and ATF4 induction in response to stress in the endoplasmic reticulum. We show that although the cells expressing mutant eIF2B genes respond normally to stress conditions by reduced global translation rates, they exhibit significantly greater increase in ATF4 induction compared to normal controls despite equal levels of stress and activity of the upstream eIF2alpha kinase. This heightened stress response observed in primary fibroblasts that suffer from minor loss of basal eIF2B activity may be employed as an initial screening tool for CACH/VWM leukodystrophy. PMID- 16041585 TI - Molecular cloning of LILRC1 and LILRC2 in the mouse and the rat, two novel immunoglobulin-like receptors encoded by the leukocyte receptor gene complex. AB - We report the molecular cloning of two novel single-member receptor families with homology to LILR/CD85, PIR, and gp49: LILRC1 in the rat and the mouse, and LILRC2 in the rat. LILRC1 and LILRC2 both have two extracellular Ig-like domains and a cytoplasmic tail devoid of any known signaling motifs. The transmembrane regions of LILRC1 and LILRC2 contain an arginine residue, a common feature in receptors that associate with activating adaptor proteins. Rat and mouse LILRC1 are orthologs sharing 81.5% amino acid identity. LILRC2 represents a distinct receptor family, 47.9% identical to LILRC1. No murine LILRC2 ortholog was detected in genome or expressed sequence tag sequence databases. By radiation hybrid mapping, the rat Lilrc1 and Lilrc2 loci were localized to the leukocyte receptor gene complex (LRC) on chromosome 1, and the mouse Lilrc1 locus was mapped to the LRC on chromosome 7. Moreover, the mouse and rat Lilrc1 loci were localized to similar positions within the LRC. As shown by RT-PCR, rat LILRC1 was expressed by B cells, neutrophils, and a macrophage cell line. Transcription of LILRC2 was detected in T cells, B cells, neutrophils, and macrophages. PMID- 16041586 TI - Leber's hereditary optic neuropathy and the pupil. PMID- 16041587 TI - Periorbital necrotising fasciitis after minor trauma. AB - INTRODUCTION: Necrotising fasciitis or streptococcal gangrene is a rare and often fatal soft tissue infection usually affecting the limbs and trunk. Facial involvement is exceedingly rare due to the excellent blood supply of this region. METHODS: We report a case of initially misdiagnosed streptococcal gangrene of the eyelids precipitated by minor trauma and which progressed despite intensive medical therapy. RESULTS: A 53-year-old man with a history of alcohol abuse developed rapidly increasing left-sided periorbital oedema, erythema and skin vesicles soon after sustaining a laceration to his left upper lid. It was initially treated as herpes zoster ophthalmicus complicated by a secondary bacterial cellulitis. Bacterial cultures grew group A beta haemolytic Streptococcus pyogenes. Despite 8 days of high-dose parenteral antibiotic therapy and oral acyclovir, characteristic blisters formed and necrosis of the periorbital skin and subcutaneous tissues ensued. Surgical debridement was performed and the fasciitis rapidly resolved. CONCLUSION: Physicians and ophthalmologists must be aware of the risk factors, although rare, for periorbital necrotising fasciitis and the cardinal signs that differentiate this condition from common non-necrotising preseptal cellulitis. Prompt recognition and early surgical debridement are crucial in limiting the morbidity and mortality from severe forms of this disease. PMID- 16041588 TI - Massive hemorrhagic retinal detachment during radial optic neurotomy. AB - BACKGROUND: We encountered obvious arterial bleeding from the incision site during radial optic neurotomy (RON) in a 55-year-old woman with central retinal vein occlusion (CRVO) and report the findings herein. CASE REPORT: The patient initially demonstrated a retinal hemorrhage and macular edema due to a left CRVO. Her corrected visual acuity was 0.5 OS. Since her symptoms did not improve and were aggravated, even though she received an oral anticoagulant drug orally, RON was performed 2 months after the onset of symptoms. During surgery, obvious arterial bleeding started from the RON incision site and it was not arrested although the infusion bottle was fully elevated. However, a subsequent infusion of liquid perfluoro-carbon successfully stopped the bleeding by direct pressure. Hemorrhagic retinal detachment occurred in the nasal quadrant and postoperative vitreous hemorrhage was treated surgically. The hemorrhagic retinal detachment was gradually absorbed and replaced by fibrous scar tissue. Two years after the surgery, no expansion of retinal detachment has been observed and corrected visual acuity is being maintained at 0.2 OS. CONCLUSION: We conclude that RON for CRVO carries a risk of arterial bleeding and that infusion of liquid perfluoro carbon seems to be effective to arrest arterial bleeding from the RON site. PMID- 16041589 TI - Ocular pulse amplitude after trabeculectomy. AB - BACKGROUND: The ocular pulse amplitude (OPA) is the difference between the minimum and maximum values of the pulsatile intraocular pressure (IOP) wave contour. The OPA depends on ocular perfusion and IOP, which are both affected by a trabeculectomy (TE). The aim of this study was to investigate how the OPA changes after TE and whether an early change in OPA can be used as a prognostic marker for a successful long-term outcome. METHODS: Fourteen consecutive patients (26-84 years old) with medically uncontrolled primary open-angle or pseudoexfoliation glaucoma were included in the study. IOP and OPA were measured with a dynamic contour tonometer before and after TE on days -1, +1, +7, +14, +21, +28, +42, +56, +70, and +84. The OPA of the contralateral eye was used to control for variations in systemic haemodynamics. TE was regarded as successful if a persistent drop in IOP of at least 20% without the use of IOP-lowering treatment was achieved. Data were analysed using receiver operating characteristic curves, Kaplan-Meier survival curves and Mann-Whitney two-sample analysis. RESULTS: Five out of 14 TEs had an entirely successful outcome. The other 9 patients required additional interventions such as suturolysis, needling of subconjunctival scar tissue and antimetabolite injections during the 3-month period after the TE. On the first day after surgery, OPA decreased in 12 patients and increased in 2 patients compared with the preoperative measurements. In the 5 patients with a successful long-term outcome, OPA dropped by 3.38+/-1.79 mmHg (mean+/-SE), whereas the initial OPA drop in those cases that required additional interventions was 0.62+/-1.81 mmHg only (p<0.01). IOP dropped by 13.10+/-2.14 mmHg in the successful group and by 5.84+/-2.51 mmHg in the unsuccessful group (p=0.19). Kaplan-Meier estimates of survival showed that patients with an initial OPA drop of more than 2.0 mmHg had a significantly better chance of an entirely uncomplicated 3-month outcome after TE than patients with an initial OPA drop of less than 2.0 mmHg (log rank p<0.01). CONCLUSION: This pilot study indicates that an early drop in OPA of more than 2.0 mmHg after TE may be a good prognostic parameter for successful long-term control of IOP. PMID- 16041590 TI - Impact of 2-deoxy-2[F-18]fluoro-D-glucose Positron Emission Tomography on the management of patients with advanced melanoma. AB - OBJECTIVES: Accurate staging of patients with melanoma is vital to guide appropriate treatment. 2-Deoxy-2-[F-18]fluoro-D-glucose (FDG)-positron emission tomography (PET) has been reported to be a sensitive and specific technique for the staging of advanced melanoma, however, few studies provide information regarding its impact on patient management. METHODS: We retrospectively reviewed the FDG-PET scan results of 92 patients with melanoma who had 126 scans performed over a six-year period. These patients were seen at the specialist melanoma clinic at our Institution, and 84 patients (92%) had stage III or IV disease. FDG PET scan results were correlated with computed tomography (CT) scans and other imaging when available, and with clinical follow-up of a minimum of three to six months. The impact of FDG-PET scans on patient management was also assessed. RESULTS: On a lesion-by-lesion analysis, FDG-PET had a sensitivity of 92%, a specificity of 88%, and an accuracy of 91%. FDG-PET correctly affected the clinical decision-making process in 40 of 126 patient studies (32%), particularly assisting in the selection of patients for surgery. CONCLUSION: FDG-PET has an important role in guiding the management of patients with advanced melanoma, particularly when surgery is contemplated. PMID- 16041591 TI - Comparison of [14C]FMAU, [3H]FEAU, [14C]FIAU, and [3H]PCV for monitoring reporter gene expression of wild type and mutant herpes simplex virus type 1 thymidine kinase in cell culture. AB - PURPOSE: To assess the optimal reporter probe/reporter gene combination for monitoring herpes simplex virus type 1 thymidine kinase (HSV1-tk) gene expression, we compared the cellular uptake of 1-(2'-fluoro-2'-deoxy-D arabinofuranosyl)-5-methyluracil (FMAU), 2'-fluoro-2'-deoxyarabinofuranosyl-5 ethyluracil (FEAU), 2'-fluoro-2'-deoxy-beta-D-arabinofuranosyl-5-iodouracil (FIAU) and penciclovir (PCV) in both HSV1-tk and HSV1-sr39tk expressing cells. PROCEDURES: For stably transfected cell studies, C6 rat glioma cells, C6 HSV1-tk transfectant, C6 mutant HSV1-sr39tk transfectant, rat Morris hepatoma cells (MH3924A), and MH3924A HSV1-tk transfectant cells were used. For adenoviral infection studies, C6 rat glioma cells were exposed to serial titers of AdCMV HSV1-tk, AdCMV-HSV1-sr39tk, or AdCMV-fluc for 24 hours. These cells were incubated with [(14)C]FMAU, [(3)H]FEAU, [(14)C]FIAU, and [(3)H]PCV, and cellular uptake of radioactivity was measured. RESULTS: [(3)H]FEAU exhibited the highest or second highest accumulation and the most selectivity regardless of the mode of gene transfer for both HSV1-tk and mutant HSV1-sr39tk reporter genes. CONCLUSION: This combination of high accumulation and high selectivity for both HSV1-tk and HSV1-sr39tk makes suitably radiolabeled FEAU a promising candidate as a radiotracer for imaging HSV1-tk/HSV1-sr39tk gene expression in living subjects. PMID- 16041592 TI - Higher lesion conspicuity for SENSE dynamic MRI in detecting hypervascular hepatocellular carcinoma: analysis through the measurements of liver SNR and lesion-liver CNR comparison with conventional dynamic MRI. AB - The aim of our study was to compare the signal-to-noise ratio (SNR) of liver parenchyma and the contrast-to-noise ratio (CNR) of hypervascular hepatocellular carcinoma (HCC) between conventional and SENSE dynamic MRI. Thirty-one consecutive patients who were strongly suspected of having HCC were enrolled in our study. The subjects consisted of 20 men and 11 women aged 52 years to 79 years (mean 66.8 years). Dynamic MRI was performed for each patient, with SENSE (SENSE MRI) and without SENSE (conventional MRI) on separate days. For the quantitative analysis, the liver SNR and the lesion-liver CNR of 25 hypervascular HCCs detected on both conventional and SENSE dynamic MRI were measured. The liver SNR of the arterial phase and the portal venous phase was 84.1+/-24.7 and 104.7+/ 34.3, respectively, in conventional MRI, while it was 62.9+/-19.5 and 44.5+/ 18.2, respectively, in SENSE MRI. SENSE MRI showed a statistically significantly lower SNR than conventional MRI (P<0.01). The lesion-liver CNR was 26.3+/-15.9 in conventional MRI and 39.0+/-19.6 in SENSE MRI. The lesion-liver CNR in SENSE MRI was significantly higher than in conventional MRI (P<0.01). The SNR in SENSE MRI is significantly lower than in conventional MRI, although the lesion CNR is significantly higher than in conventional MRI. PMID- 16041593 TI - The vertically orientated fibular collateral ligament: a secondary sign of anterior cruciate ligament rupture on magnetic resonance imaging. PMID- 16041594 TI - Gender differences in renal tubular taurocholate transport. AB - The bile acids filtered through the glomeruli nearly completely escape urinary excretion due to an efficient tubular reabsorption process. Reabsorption is mediated by the sodium-dependent bile acid transporter ASBT, which is localized in the brush border membranes of proximal tubular cells. The purpose of the present study was to assess whether tubular taurocholate transport is regulated by sex hormones. Clearance studies and studies on proximal tubular cells freshly isolated from rat kidneys were performed. The studies with the isolated proximal tubular cells revealed a cell to bath 3H-taurocholate accumulation ratio of 5.63+/-0.28 in male and of 3.67+/-0.43 in female rats (p<0.01). This difference in cellular taurocholate uptake was corroborated by the clearance studies, which showed a 3H-taurocholate clearance of 133.9+/-28.1 in male rats and of 262.0+/ 45.4 microl/min x 100 g b.w. in female rats (p<0.05). Testosterone treatment of female rats did not significantly alter the cell to bath 3H-taurocholate accumulation ratio. However, the cellular taurocholate accumulation significantly decreased, by 61.6+/-10.1%, following ethinylestradiol treatment of male rats. Ovariectomy, chemical castration of female rats with buserelin or treatment of female rats with the estrogen receptor antagonist ICI 182780 did not affect taurocholate uptake, but treatment of ovariectomized rats with ethinylestradiol decreased the taurocholate accumulation ratio by 53.7+/-15.8%. By determination of serum bile acids the possibility was excluded that this change was an indirect effect of cholestasis induced by ethinylestradiol. This study demonstrates gender differences in the renal handling of taurocholate in rats that may be related to an inhibitory effect of estrogens on taurocholate transport in proximal tubular cells. Since the ASBT protein content of the proximal tubular cells was found not to be different between male and female rats, a nongenomic mechanism may underly this estrogen effect. PMID- 16041595 TI - Effects of cinnarizine on calcium and pressure-dependent potassium currents in guinea pig vestibular hair cells. AB - In vestibular hair cells, K+ currents induced by rises in hydrostatic pressure have recently been demonstrated. These currents are inhibited by charybdotoxin, a blocker of Ca2+-dependent K+ channels. On the other hand, cinnarizine is a blocker of voltage-gated Ca2+ currents in hair cells and is used as a drug in conditions with vestibular vertigo. Our aim was to test in patch-clamp experiments (conventional whole-cell mode) whether cinnarizine, by reducing Ca2+ influx, inhibited Ca2+ and pressure-sensitive K+ currents in vestibular type-II hair cells of guinea pigs. A quantitatively similar inhibition of K+ currents was evoked by extracellular Ca2+ removal, cinnarizine (0.5 microM), and the L-type Ca2+ channel blocker nifedipine (3 microM). Cinnarizine abrogated increases of K+ currents induced by increases in the hydrostatic pressure (from 0.2 to 0.5 cm H2O). At a higher concentration (1 microM), cinnarizine elicited K+ current inhibitions larger than those elicited by Ca2+ removal. Moreover, it reduced K+ currents in the absence of Ca2+, in contrast to nifedipine. However, charybdotoxin abolished these effects of cinnarizine. We thus conclude that cinnarizine inhibits, by two mechanisms, pressure-induced currents that are sensitive to charybdotoxin and Ca2+. It reduces Ca2+ influx and exerts a Ca2+ independent inhibition, with a lower IC50 than that required for Ca2+ channel blockade. These two actions may importantly contribute to its therapeutic effects. PMID- 16041596 TI - Comparison of information on the pharmacokinetic interactions of Ca antagonists in the package inserts from three countries (Japan, USA and UK). AB - OBJECTIVE: Ca antagonists are one of the most popular classes of drugs used to treat hypertension and angina. These drugs may interact with either CYP3A4 or MDR 1 substrates, with the degree of interaction differing with each drug. We carried out a literature search to examine and compare the extent to which crucial pharmacokinetic (PK) information is included in package inserts (PIs) in Japan, USA and the UK. METHODS: A MEDLINE search from 1966 to November 2004 was undertaken with the aim of identifying studies on clinical PK drug interactions between seven Ca antagonists that are available in three countries and three CYP3A4 inhibitors (erythromycin, itraconazole and cimetidine), a CYP3A4 inhibitory food, grapefruit juice (GFJ) and the MDR-1 substrate, digoxin. The current PIs for Ca antagonists were obtained from the website of the regulatory authorities or the electronic Medicines Compendium. RESULTS: Of all possible combinations of seven Ca antagonists with three CYP3A4 inhibitor drugs, drug interaction information was available in the literature on nine combinations: Seven of these were listed in the USA PIs, two in the UK PIs, and none in the Japanese PIs. Interaction studies with GFJ were reported for every Ca antagonist; PIs in the USA provided quantitative data for four of these interactions, whereas UK PIs provided quantitative data for only one of the interactions and Japanese PIs provided no quantitative information. The PK data of co-medication of digoxin with Ca antagonists have been reported for every Ca antagonists. The USA PIs provided quantitative data for five Ca antagonists, whereas the UK PIs provided quantitative data for three Ca antagonists and Japanese PIs provided no quantitative data. CONCLUSION: The literature search revealed that PIs in the USA provided a great deal of quantitative information on PK interactions between Ca antagonists and other drugs or GFJ. In contrast, PIs in the UK and Japan did not provide sufficient information. We conclude that crucial quantitative information on these drug interactions should be incorporated in PIs, especially in Japan and the UK, as a means of assisting healthcare providers. PMID- 16041597 TI - The pharmacokinetics and pharmacodynamics of atovaquone and proguanil for the treatment of uncomplicated falciparum malaria in third-trimester pregnant women. AB - OBJECTIVE: To investigate the pharmacokinetics, safety and efficacy of the recommended 3-day treatment regimen of Malarone in third-trimester pregnant women with acute uncomplicated falciparum malaria. METHODS: Twenty-six pregnant women in their third trimester (gestational age: 24-34 weeks) with acute uncomplicated Plasmodium falciparum malaria who fulfilled the enrollment criteria were recruited from the antenatal clinics of Mae Sot Hospital, Tak Province, Thailand, (n = 8) and the Tropical Diseases Research Centre, Ndola, Zambia (n = 18). Patients were treated with four Malarone tablets (GlaxoSmithKline: each tablet contains 250 mg atovaquone and 100 mg proguanil) once daily for 3 consecutive days. Blood samples were taken for pharmacokinetic investigations of atovaquone, proguanil, and cycloguanil up to 288 h (day 14) after the last dose. Urine samples were collected for the evaluation of proguanil and cycloguanil 0-8, 8-16, 16-24 and 24-48 h after the last dose. Efficacy assessments included the clinical and parasitological evaluation of mothers and newborns. Adverse events were evaluated at each visit to the antenatal clinics. RESULTS: Malarone appeared to be effective and well tolerated when used for the treatment of falciparum malaria in pregnant women. All patients showed prompt clinical improvement and the disappearance of parasitaemia after treatment. There were no serious adverse effects or unexpected adverse effects and no stillbirths or spontaneous abortions. The plasma concentration-time profiles of atovaquone and proguanil in most cases were best characterised by the two-compartment open model with zero order input with/without absorption lag time and first-order elimination. There were no significant differences in any of the pharmacokinetic parameters of atovaquone, proguanil or cycloguanil between patients from Thailand and Zambia. For atovaquone, a Cmax of 1.33-8.33 microg/ml was reached at 2.0-9.3 h after the last dose on day 2. V/F, CL/F and t(1/2beta) were 6.9-39.5 l/kg, 83-384 ml/h/kg, and 57.8-130.8 h, respectively. The Cmax and t(max) values for proguanil versus cycloguanil were 383-918 versus 0-129 ng/ml and 3.3-8.6 versus 3-12 h, respectively. V/F, CL/F, and t(1/2beta) values for proguanil were 10.7-34.0 l/kg, 431-1,662 ml/h/kg and 11.2-30.3 h. The CL(R-CG), t(1/2z), (CG), proguanil/cycloguanil metabolic ratios, AUC ratios for proguanil to cycloguanil (AUC(PG/CG)) were 107.2-1,001 ml/h/kg, 5-95 ml/h/kg, 7.8-20.7 h, 5-57, and 4.7 20.2, respectively. CONCLUSION: The pharmacokinetics of atovaquone and cycloguanil appeared to be influenced by the pregnancy status, resulting in an decrease in the Cmax and AUC of approximately twofold. PMID- 16041598 TI - Steady-state pharmacokinetics of atazanavir given alone or in combination with saquinavir hard-gel capsules or amprenavir in HIV-1-infected patients. AB - OBJECTIVE: The aim of this pilot study was to examine the pharmacokinetics of atazanavir (ATV) when given in combination with amprenavir (APV) or saquinavir hard-gel capsules (SQV) to human immunodeficiency virus (HIV)-positive patients. METHODS: Included in the study were 34 HIV-infected patients enrolled in the ATV Early Access Program, who were treated with unboosted ATV alone (group 1) or with the double protease inhibitor combinations, ATV plus APV (group 2) or ATV plus SQV (group 3). ATV was given at a daily dose of 400 mg q.d. with the morning meal with SQV 1200 mg per day or APV 1200 mg per day. Serial blood samples for steady state ATV pharmacokinetics were collected before the morning dose and at 1, 2, 3, 6, 8 and 24 h post-dosing. ATV plasma concentrations were measured using a high performance liquid chromatography method with ultraviolet detection. RESULTS: Of the patients, 12 received ATV as a single protease inhibitor; 12 received ATV in combination with APV; and 10 in combination with SQV. Geometric mean (coefficient of variation) ATV C(trough) was 110 ng/ml (2.38), 86 ng/ml (0.84) and 149 ng/ml (2.01) in groups 1, 2 and 3, respectively. ATV C(trough) in both double protease inhibitor combination regimens was not significantly different from that as a single protease inhibitor [geometric mean ratio (GMR): 0.77; 95% confidence interval (CI): 0.38-1.58, P=not significant for group 2 versus group 1 and 1.34, 0.40-4.49, P=not significant, for group 3 versus group 1). Patients treated with ATV plus APV had a 40.2% lower ATV C(max) and a 30.8% smaller ATV AUC than the reference group treated with unboosted ATV alone: both these differences were statistically significant (GMR, 95% CI: 0.59, 0.41-0.85, P=0.005 and 0.69, 0.48 0.99, P=0.056, respectively). No difference was observed for either C(max) or AUC between the group treated with ATV plus SQV and the reference group (GMR, 95% CI: 0.78, 0.47-1.30, P=not significant and 1.24, 0.73-2.10, P=not significant, respectively). CONCLUSION: ATV pharmacokinetics does not seem to be influenced by the concomitant administration of SQV, whereas APV significantly lowers plasma ATV levels. PMID- 16041599 TI - MDMA (ecstasy) pharmacokinetics in a CYP2D6 poor metaboliser and in nine CYP2D6 extensive metabolisers. PMID- 16041600 TI - Embryonic and postnatal development of masticatory and tongue muscles. AB - This review summarizes findings concerning the unique developmental characteristics of mouse head muscles (mainly the masticatory and tongue muscles) and compares their characteristics with those of other muscles. The developmental origin of the masticatory muscles is the somitomeres, whereas the tongue and other muscles, such as the trunk (deep muscles of the back, body wall muscles) and limb muscles, originate from the somites. The program controlling the early stages of masticatory myogenesis, such as the specification and migration of muscle progenitor cells, is distinctly different from those in trunk and limb myogenesis. Tongue myogenesis follows a similar regulatory program to that for limb myogenesis. Myogenesis and synaptogenesis in the masticatory muscles are delayed in comparison with other muscles and are not complete even at birth, whereas the development of tongue muscles proceeds faster than those of other muscles and ends at around birth. The regulatory programs for masticatory and tongue myogenesis seem to depend on the developmental origins of the muscles, i.e., the origin being either a somite or somitomere, whereas myogenesis and synaptogenesis seem to progress to serve the functional requirements of the masticatory and tongue muscles. PMID- 16041602 TI - Trace analysis of polycyclic aromatic hydrocarbons in suspended particulate matter by accelerated solvent extraction followed by gas chromatography-mass spectrometry. AB - An analytical procedure based on extraction by accelerated solvent extraction (ASE) followed by gas chromatography-mass spectrometry (GC/MS) analysis has been developed for the determination of particulate polycyclic aromatic hydrocarbons (PAHs) from large-volume water samples (20 L). The effect of temperature and number of cycles on the efficiency of ASE was investigated: the best results were obtained by using a temperature of 100 degrees C and one static cycle. A mixture of hexane/acetone 1:1 (v/v) was used as extraction solvent. Mean total method recovery under optimized conditions was 85%. The developed methodology was applied to the analysis of suspended particulate matter from Lake Maggiore waters (north of Italy). Mean PAH concentrations in suspended particulate matter from Lake Maggiore ranged from 0.2 ng L(-1) for anthracene to 18.7 ng L(-1) for naphthalene. PMID- 16041601 TI - No correlation between the p38 MAPK pathway and the contractile dysfunction in diabetic cardiomyocytes: hyperglycaemia-induced signalling and contractile function. AB - Besides the classical cardiovascular diseases, high levels of blood glucose directly interfere with cardiomyocytes. The mechanisms responsible for this have not yet been explored in detail. This study aims to determine if hyperglycaemia has any impact on prominent signalling molecules and on the contractile function of cardiomyocytes. Freshly isolated cardiomyocytes from adult rats were treated with various concentrations of glucose. Formed free radicals were measured by DCF fluorescence. TGFbeta expression and p38 MAP-kinase (MAPK) activation were measured by Western blotting. The contractile efficiency was determined by measurement of the maximal amount of cell shortening. Glucose (30 mM) caused an increase in formation of radicals, phosphorylation of p38 MAPK, and TGFbeta expression. Under conditions of low viscosity (1 cp), contractile responses to hyperglycaemia (15 mM) were not altered in contrast to control. However, enhancement of viscosity (400 cp) effected a limitation of contractile function. The responsiveness to beta-adrenoceptor stimulation did not change. Neither inhibition of p38 MAPK with SB 202190 (1 microM) nor inhibition of reactive oxygen species with vitamin C did alter these measured functional parameters. Diabetes mellitus directly influences the activation degree of prominent signalling molecules and the contractile function of adult ventricular cardiomyocytes, which results in facilitating in the development of diabetic cardiomyopathy. PMID- 16041603 TI - In vitro monitoring of GTPase activity and enzyme kinetics studies using capillary electrophoresis. AB - A capillary electrophoresis (CE)-based method for the in vitro detection and monitoring of nucleotide-triphosphatase activity is described. This robust and reproducible method was used to investigate GTPase activity of a recombinant protein construct containing the catalytic domain of Human SEPT4/Bradeion beta (GST-rDGTPase). This example application demonstrates that the CE technique can replace classical radioactive methods for GTPase activity assays and may be used as a routine analytical tool. Enzyme kinetics of GST-rDGTPase was studied and yielded the following kinetic parameters: v(max) = 1.7 microM min(-1) +/- 0.1, Km = 1.0 mM +/- 0.3, and apKcat = 9 x 10(-3) s(-1). In addition the effect of co factors such as Mg2+ and Mn2+ on the catalytic activity was investigated. The described analytical method was also shown to be useful to analyze diphosphated and triphosphated forms of other nucleotides. PMID- 16041604 TI - Detection of transferrin isoforms in human serum: comparison of UV and ICP-MS detection after CZE and HPLC separations. AB - Two methods for separation of transferrin (Tf) sialoforms, capillary electrophoresis (CE) and high performance liquid chromatography (HPLC) with conventional UV absorbance detection, have been investigated and compared. First, conditions affecting the separation of the Tf isoforms by capillary zone electrophoresis and HPLC were carefully optimized. The use of 15 mmol L(-1) borate buffer (pH 8.4) containing 3 mmol L(-1) diaminobutane (DAB) as additive enabled good separation of the Tf isoforms by CE (75 cm x 50 microm i.d. fused silica capillary) at 25 kV. In HPLC, a gradient of ammonium acetate (from 0 to 250 mmol L(-1) in 45 min) buffered at pH 6 (Tris-HCl) proved suitable for separation of Tf isoforms on a Mono-Q HR 5/5 anion-exchange column. On-line specific detection of the iron associated with the different Tf isoforms, after Fe saturation, by inductively coupled plasma mass spectrometry (ICP-MS) was studied in detail to compare its analytical performance with UV detection. For both CE and HPLC an octapole reaction system (ORS) ICP-MS instrument was used to minimize polyatomic interferences on the (56)Fe major isotope. Limits of detection of the different isoforms were in the range of 0.02-0.04 micromol L(-1) Tf for HPLC-ICP (ORS)-MS. This hybrid technique proved more selective and reliable detection of transferrin isoforms with 2, 3, 4, 5, and 6 sialic acid residues (S(2), S(3), S(4), S(5), and S(6)) in real serum samples. Interesting results from iron speciation of Tf in serum from healthy individuals and from pregnant women are given. PMID- 16041605 TI - A phase response curve for circannual rhythm in the varied carpet beetle Anthrenus verbasci. AB - We know that entrainment, a stable phase relationship with an environmental cycle, must be established for a biological clock to function properly. Phase response curves (PRCs), which are plots of phase shifts that result as a function of the phase of a stimulus, have been created to examine the mode of entrainment. In circadian rhythms, single-light pulse PRCs have been obtained by giving a light pulse to various phases of a free-running rhythm under continuous darkness. This successfully explains the entrainment to light-dark cycles. Some organisms show circannual rhythms. In some of these, changes in photoperiod entrain the circannual rhythms. However, no single-pulse PRCs have been created. Here we show the PRC to a long-day pulse superimposed for 4 weeks over constant short days in the circannual pupation rhythm in the varied carpet beetle Anthrenus verbasci. Because the shape of that PRC closely resembles that of the Type 0 PRC with large phase shifts in circadian rhythms, we suggest that an oscillator having a common feature in the phase response with the circadian clock, produces a circannual rhythm. PMID- 16041606 TI - Biological monitoring of carbon disulphide and phthalate exposure in the contemporary rubber industry. AB - OBJECTIVES: We studied the range in urinary levels of 2-thiothiazolidine-4 carboxyl acid (TTCA), a metabolite of CS2 and phthalic acid (PA), a common metabolite of phthalates, across factories and departments in the contemporary rubber manufacturing industry. METHODS: Spot urine samples from 101 rubber workers employed in nine different factories were collected on Sunday and during the workweek on Tuesday, Wednesday and Thursday at approximately 4 pm. In total, 386 urine samples were successfully analyzed. RESULTS: Levels of both biomarkers increased significantly by a factor 2 (paired t-test P-value <0.05) during the working week as compared to the Sunday biomarker levels with absolute increases of approximately 70 microg/l and 5 micromol/mol creatinine for PA and TTCA, respectively. Levels in both biomarkers did not differ markedly between working days. Increases seemed to be restricted to specific factories and/or departments (e.g. molding and curing). CONCLUSIONS: The results of this study demonstrate that rubber workers in the contemporary rubber industry are exposed to phthalates and low levels of CS2 ( approximately 0.05 ppm) as measured by PA and TTCA, respectively. Exposures to both compounds are largely driven by specific circumstances in factories. Therefore, when estimating exposures to phthalates and CS2 detailed information should be collected on the type and amount of phthalate containing ester plasticizers, dithiocarbamates and thiurams used. Preferably, personal exposure data should be collected. In this case, biological monitoring seems a reasonable approach. However, in the case of PA attention should be given to individual background levels as this could lead to a substantial overestimation of the occupational contribution to total phthalate exposure. PMID- 16041607 TI - Contribution of combined colposcopy and cytology in cervical pathology. AB - AIM: The regular Papanicolaou (Pap) smear is the cornerstone of women's preventive healthcare. The introduction of the regular Pap smear as a screening tool for cervical cancer has markedly decreased the number of deaths from cervical cancer. During the past decade, however, the rate of death from cervical cancer has remained relatively static. This screening method is known to have a high percentage of false negative results. To improve the detection of cervical lesions using the Pap smear in screening, a number of adjunct procedures have been developed. The purpose of this study is to evaluate the utility of a magnified chemoluminescent screening examination (Colposcopy) combined with the Pap smear in detecting cervical abnormalities. METHODS: We investigated a cohort of 58 subjects who have been forwarded for colposcopic evaluation due to referral cytology suggestive of persistent inflammatory process not otherwise specified, and cervical intraepithelial or invasive neoplasia, in Chania Colposcopic clinic. RESULTS: The higher the lesion detected by cytology, the more severe the corresponding colposcopic impression viewed. CONCLUSION: The data presented here are in harmony with previous reports and share our experience in a Regional Community Hospital Colposcopy Clinic. An integrated cytology-colposcopy program facilitates the assessment and identification of women harboring cervical pathological conditions. PMID- 16041608 TI - Cysteine protease falcipain 1 in Plasmodium falciparum is biochemically distinct from its isozymes. AB - Falcipains form a class of papain-like cysteine proteases found in Plasmodium falciparum. This group of proteases has been suggested to be promising targets for anti-malarial chemotherapy. Despite being the first falcipain to be identified, the physiological role(s) of falcipain 1 (fp1) remains a mystery. Its suggested functions include haemoglobin degradation, erythrocytic invasion and oocyst production. In this study, the procurement of the gene coding for fp1 and its soluble expression in a heterologous host, Escherichia coli, have enabled further enzyme characterization. The recombinant fp1 protease was found to be unlike falcipain 2 (fp2A) in being more active at neutral pH than at acidic pH against the Z-LR-AMC fluorogenic substrate, suggesting a probable localization in the cytosol and not in the food vacuole. Interestingly, a common cysteine specific inhibitor, E64, did not inhibit fp1 activity, indicating dissimilar biochemical characteristics of fp1 from the other falcipains. This may be explained by computational analysis of the primary structures of the falcipain isozymes, as well as that of papain. The analysis revealed that Tyr61 (papain numbering), which is correspondingly absent in fp1, might be an important residue involved in E64 substrate binding. PMID- 16041609 TI - Rare causes of gastric outlet obstruction in children. AB - Gastric outlet obstruction (GOO) presenting beyond the newborn period is a relatively rare condition, when infantile hypertrophic pyloric stenosis (IHPS) is excluded. This report describes the clinical features, evaluation and management of 18 patients with GOO which was not caused by IHPS. The sex, age, and main presenting features were recorded on admission. Hemoglobin estimation, serum chemistry and blood gas analysis were also assessed in these patients. The diagnosis was confirmed with gastrointestinal barium, B ultrasound, gastroscopy, or at surgery. Some of them underwent gastroscopy with biopsy sampling for rapid urease test or histologic examination with Giemsa stains for Helicobacter pylori. Thirteen patients had peptic ulcer disease, of which six were successfully treated with medications. While the other seven needed operative management which included Heineke-Mikulicz pyloroplasty, gastroduodenostomy or gastrojejunostomy, respectively. All the patients who had Heineke-Mikulicz pyloroplasty or gastrojejunostomy developed bile reflux gastritis that could be alleviated with omeprazol. None of the patients developed dumping syndrome, failing to thrive, or anemia. Three children with antral diaphragm recovered without any complications by diaphragm excision with Heineke-Mikulicz pyloroplasty. The pylorus was compressed by fibrotic band in one 30-month boy, and he recovered uneventfully with Heineke-Mikulicz pyloroplasty during 10-year follow-up. Gastric outlet obstruction secondary to ingestion of sulfuric acid was noted in one patient, and he was successfully treated with gastroduodenostomy. Our data suggests that the satisfactory results could be expected in children with GOO with different therapeutic modalities based on the specific cause and degree of obstruction. PMID- 16041610 TI - Gemcitabine (GEM) plus oxaliplatin, folinic acid, and 5-fluorouracil (FOLFOX-4) in patients with advanced gastric cancer. AB - BACKGROUND AND AIMS: oxaliplatin in combination with folinic acid (FA) and infusional 5-fluorouracil (5-FU) has shown significant anti-tumor activity in gastric cancer patients (FOLFOX). Previous studies have shown that gemcitabine (GEM), a new fluorinated anti-metabolite, enhances the individual anti-tumor activity of either 5-FU or oxaliplatin. We have therefore designed a multi-center phase II trial in order to test a novel GEM+FOLFOX-4 regimen in patients with metastatic gastric cancer. METHODS: we enrolled 36 patients, 28 males and 8 females, with an average age of 64.4 years (range 37-78), who received bi-weekly treatment with GEM (1,000 mg/m2 on day 1), levo-FA (100 mg/m2 on days 1 and 2), a 5-FU (400 mg/m2) bolus injection followed by 22-h continuous infusion (800 mg/m2) on days 1 and 2, and oxaliplatin 85 mg/m2 in a 4-6 h intravenous (i.v.) infusion before the second FUFA administration on day 2. RESULTS: the most frequent side effect was grade 1-2 hematological toxicity and late sensorial neurotoxicity. Two patients developed hypersensitivity to oxaliplatin while another developed an aseptic eosinophilic pneumonitis. Two patients refused to continue the treatment after two cycles of chemotherapy and were lost at the follow-up. Among the remaining 34 patients four achieved a complete response, 15 a partial response, 12 had a stable disease and three progressed. CONCLUSIONS: these results may grant the rationale to evaluate this multi-drug combination in randomized phase III trials in advanced gastric cancer. PMID- 16041611 TI - A phase I study to determine the effect of tamoxifen on the pharmacokinetics of a single 250 mg oral dose of gefitinib (IRESSA) in healthy male volunteers. AB - OBJECTIVES: To determine the effect of tamoxifen on the pharmacokinetics of a single 250 mg oral dose of gefitinib (IRESSA) in healthy volunteers. METHODS: An open-label, single-center, phase I study in healthy male volunteers. Each volunteer received a single 250 mg oral dose of gefitinib on day 1. On days 11 14, oral loading doses of 60 mg tamoxifen were administered, followed by 20 mg tamoxifen for a further 16 days to maintain steady-state exposure. On day 24, volunteers received a second single 250 mg oral dose of gefitinib. The last dose of tamoxifen was given on day 30. Pharmacokinetic and safety assessments were conducted throughout the trial. RESULTS: A total of 18 volunteers were recruited. The presence of tamoxifen did not have a clinically significant effect on the primary variables AUC and Cmax of gefitinib, nor on the secondary variables AUC(0 t), tmax, t1/2, and lambdaz. The geometric least square mean values for AUC were 3,407.6 versus 3,397.9 ng.h/ml in the absence and presence of tamoxifen, respectively (90% CL 0.894, 1.112) and for Cmax were 110.8 versus 103.6 ng/ml, respectively (90% CL 0.786, 1.111). The combination of gefitinib with tamoxifen was generally well tolerated by the volunteers. There were no serious adverse events and no volunteer discontinued the study due to an adverse event. NCI-CTC grade 1/2 drug-related adverse events were observed in seven volunteers, including loose stools and skin events associated with gefitinib, and lethargy and headache, flushing, and dizziness associated with tamoxifen. CONCLUSIONS: This study suggests that tamoxifen has no significant effect on the pharmacokinetics, tolerability, or safety of a single 250 mg oral dose of gefitinib. Therefore, in clinical investigations of this combination, no dose adjustment of gefitinib is indicated. PMID- 16041612 TI - Habitat complexity and sex-dependent predation of mosquito larvae in containers. AB - Studies in aquatic systems have shown that habitat complexity may provide refuge or reduce the number of encounters prey have with actively searching predators. For ambush predators, habitat complexity may enhance or have no effect on predation rates because it conceals predators, reduces prey detection by predators, or visually impairs both predators and prey. We investigated the effects of habitat complexity and predation by the ambush predators Toxorhynchites rutilus and Corethrella appendiculata on their mosquito prey Aedes albopictus and Ochlerotatus triseriatus in container analogs of treeholes. As in other ambush predator-prey systems, habitat complexity did not alter the effects of T. rutilus or C. appendiculata whose presence decreased prey survivorship, shortened development time, and increased adult size compared to treatments where predators were absent. Faster growth and larger size were due to predator mediated release from competition among surviving prey. Male and female prey survivorship were similar in the absence of predators, however when predators were present, survivorship of both prey species was skewed in favor of males. We conclude that habitat complexity is relatively unimportant in shaping predator prey interactions in this treehole community, where predation risk differs between prey sexes. PMID- 16041613 TI - Edge effects and intraguild predation in native and introduced centipedes: evidence from the field and from laboratory microcosms. AB - Human alteration of habitat has increased the proportion of forest edge in areas of previously continuous forest. This edge habitat facilitates invasion of exotic species into remaining fragments. The ability of native species to resist invasion varies and may depend on intrinsic variables such as dispersal and reproductive rates as well as external factors such as rate of habitat change and the density of populations of introduced species in edge habitat. We examined the distributional and competitive relationships of two members of the class Chilopoda, Scolopocryptops sexspinosus, a centipede native to the eastern US, and Lithobius forficatus, an exotic centipede introduced from Europe. We found that L. forficatus was most abundant in edge habitat and S. sexspinosus was most abundant in the interior habitat at our field sites. Although L. forficatus was present in habitat interiors at 11 of 12 sites, there was no correlation between fragment size and numbers of L. forficatus in interior habitat. The native centipede was rarely found occupying fragment edges. We used laboratory microcosms to examine potential competitive interactions and to indirectly assess prey preferences of the two species. In microcosms both species consumed similar prey, but the native centipede, S. sexspinosus, acted as an intraguild predator on the introduced centipede. Native centipedes were competitively superior in both intraspecific and interspecific pairings. Our results suggest that intraguild predation may aid native centipedes in resisting invasion of introduced centipedes from edge habitat. PMID- 16041614 TI - Fine root chemistry and decomposition in model communities of north-temperate tree species show little response to elevated atmospheric CO2 and varying soil resource availability. AB - Rising atmospheric [CO2] has the potential to alter soil carbon (C) cycling by increasing the content of recalcitrant constituents in plant litter, thereby decreasing rates of decomposition. Because fine root turnover constitutes a large fraction of annual NPP, changes in fine root decomposition are especially important. These responses will likely be affected by soil resource availability and the life history characteristics of the dominant tree species. We evaluated the effects of elevated atmospheric [CO2] and soil resource availability on the production and chemistry, mycorrhizal colonization, and decomposition of fine roots in an early- and late-successional tree species that are economically and ecologically important in north temperate forests. Open-top chambers were used to expose young trembling aspen (Populus tremuloides) and sugar maple (Acer saccharum) trees to ambient (36 Pa) and elevated (56 Pa) atmospheric CO2. Soil resource availability was composed of two treatments that bracketed the range found in the Upper Lake States, USA. After 2.5 years of growth, sugar maple had greater fine root standing crop due to relatively greater allocation to fine roots (30% of total root biomass) relative to aspen (7% total root biomass). Relative to the low soil resources treatment, aspen fine root biomass increased 76% with increased soil resource availability, but only under elevated [CO2]. Sugar maple fine root biomass increased 26% with increased soil resource availability (relative to the low soil resources treatment), and showed little response to elevated [CO2]. Concentrations of N and soluble phenolics, and C/N ratio in roots were similar for the two species, but aspen had slightly higher lignin and lower condensed tannins contents compared to sugar maple. As predicted by source-sink models of carbon allocation, pooled constituents (C/N ratio, soluble phenolics) increased in response to increased relative carbon availability (elevated [CO2]/low soil resource availability), however, biosynthetically distinct compounds (lignin, starch, condensed tannins) did not always respond as predicted. We found that mycorrhizal colonization of fine roots was not strongly affected by atmospheric [CO2] or soil resource availability, as indicated by root ergosterol contents. Overall, absolute changes in root chemical composition in response to increases in C and soil resource availability were small and had no effect on soil fungal biomass or specific rates of fine root decomposition. We conclude that root contributions to soil carbon cycling will mainly be influenced by fine root production and turnover responses to rising atmospheric [CO2], rather than changes in substrate chemistry. PMID- 16041615 TI - Patterns, causes and consequences of regional variation in the ecology and life history of a reef fish. AB - Many species vary in their ecology across their geographic ranges in response to gradients in environmental conditions. Such variation, which can influence life history traits and subsequent demography of populations, usually occurs over large spatial scales. However, describing and understanding the causes of such variation is difficult precisely because it occurs over such large spatial scales. In this study, we document spatial variation in the ecology of a common reef fish, Stegastes beebei, in the Galapagos Islands and test a number of potential causal mechanisms. The pattern resembles that seen in latitudinal variation: individuals are larger, occur in higher densities, and live longer in the coldest region of the islands than those in the warmest region. However, in this system, demography varies among regional populations separated by <150 km. Preferred nutritious algae are more available in the cold region and comprise a greater proportion of the diet of fish in this region. Per gram reproductive effort appears to be strongly related to temperature, despite differences in the gross magnitude and timing of reproduction in different regions. A model of reproductive output suggests that fish in the warmest region are allocating a greater proportion of available energy to reproduction, resulting in apparent regional life history tradeoffs. Our data suggest that regional demographic differences in S. beebei may be driven by a combination of variation in food availability and an environmentally mediated life history tradeoff. PMID- 16041616 TI - [Quality of life of patients with squamous cell carcinoma of the oral cavity]. AB - OBJECTIVE: The aim of this study was to assess the changes in quality of life during and after treatment in patients with cancer of the oral cavity. PATIENTS AND METHODS: In the period between October 1999 and September 2000, 57 patients of the Department of Craniomaxillofacial Surgery, University of Cologne, underwent surgery, radiation therapy or the combination of both for the treatment of cancer of the oral cavity. Before, during and after the therapy their quality of life was measured with two psychometric scales. RESULTS: The average loss of quality of life in the female group was less than in the male group. Younger patients suffered more than older ones did. All patients had a loss of quality of life 3 months after the beginning of the therapy. The biggest decrease was in the group of patients treated with combined therapy, and the lowest loss in the radiated group. During the assessment period of 9 months, there were significant differences between all three groups. The size of the tumor did not show any influence on the reduction of quality of life. Patients with cancer of the tongue or maxilla showed more loss of quality of life than patients with tumors located in other regions of the oral cavity. CONCLUSION: Location of the tumor, age, gender of the patient, and type of therapy influenced the quality of life, while the size of the tumor did not. PMID- 16041618 TI - Application of endoscopy to the surgical management of craniopharyngiomas. AB - OBJECTIVE: The purpose of this study was to evaluate the efficacy of a minimally invasive/endoscopic approach to craniopharyngiomas (CPGs) given that the surgical aim was a complete excision of the tumor as a single stage procedure. The endoscope can be used with both a subfrontal and a transsphenoidal approach. METHODS: This study is a retrospective review of 36 operative patients who were seen by one surgeon. All patients had attempted complete excision. RESULTS: Patients were divided into three groups according to their preoperative status. Those in group 1 had no previous treatment and fared well, although all developed postoperative diabetes insipidus. Those in group 2 had previous surgery only and also did well with repeat surgery. Those in group 3 had been treated previously with surgery and radiotherapy and, apart from a single exception, did poorly. CONCLUSION: The endoscopic, minimally invasive approach is versatile and effective in the surgical management of both first-time and repeat cases of CPG. It offers superior visualization of surrounding neurovascular structures and allows a more complete resection of tumor. It can be used for a cranial or a nasal approach to these tumors. Sadly, it failed to prevent postoperative endocrinopathy. PMID- 16041617 TI - Reduced 123I-BMIPP uptake implies decreased myocardial flow reserve in patients with chronic stable angina. AB - PURPOSE: Long-chain fatty acid (LCFA) is the main energy source for normal myocardium at rest, but in ischemic myocardium, the main energy substrate shifts from LCFA to glucose. 123I-BMIPP is a radiolabeled LCFA analog. In chronic stable angina without previous infarction, we suppose that reduced 123I-BMIPP uptake is related to the substrate shift in myocardium with decreased myocardial flow reserve (MFR). The purpose of this study was to relate 123I-BMIPP uptake to rest myocardial blood flow (MBF), hyperemic MBF, and MFR assessed with 15O-water positron emission tomography (PET). METHODS: We enrolled 21 patients with chronic stable angina without previous infarction, all of whom underwent 123I-BMIPP single-photon emission computed tomography (SPECT) and 15O-water PET. The left ventricle was divided into 13 segments. In each segment, rest MBF and hyperemic MBF were measured by PET. 123I-BMIPP uptake was evaluated as follows: score 0=normal, 1=slightly decreased uptake, 2=moderately decreased uptake, 3=severely decreased uptake, and 4=complete defect. 123I-BMIPP uptake was compared with rest MBF, hyperemic MBF, and MFR. RESULTS: The numbers of segments with 123I-BMIPP scores 0, 1, 2, 3, and 4 were 178, 40, 25, 24, and 0, respectively. The rest MBFs for scores 0, 1, 2, and 3 were 0.93+/-0.25, 0.86+/-0.21, 0.97+/-0.30, and 0.99+/ 0.37 ml/min/g, respectively. The hyperemic MBFs for scores 0, 1, 2, and 3 were 2.76+/-1.29, 1.84+/-0.74, 1.37+/-0.39, and 1.08+/-0.40 ml/min/g, respectively. The MFRs for scores 0, 1, 2, and 3 were 3.01+/-1.38, 2.20+/-0.95, 1.44+/-0.22, and 1.10+/-0.26, respectively. As 123I-BMIPP uptake declined, hyperemic MBF and MFR decreased. CONCLUSION: In chronic stable angina without previous infarction, reduced 123I-BMIPP uptake implies decreased MFR. PMID- 16041619 TI - Enhancement of natural killer cells and increased survival of aging mice fed daily Echinacea root extract from youth. AB - In spite of Echinacea-based products being among the best-selling herbs in the world to date, to allay assorted ailments, the debate is still on-going with respect to the efficacy of ingesting the herb intermittently, continuously, or only at the beginning of an affliction. We sought, therefore, to find out if mice, receiving dietary Echinacea daily, throughout life, from youth until late middle-age, demonstrated any longevity/survival differences, and/or any differences in their various populations of immune/ hemopoietic cells. Sustained and/or high levels of these cells are crucial for longevity. Some mice were maintained on a regular chow diet to which was added Echinacea purpurea daily (2 mg/mouse), from puberty (7 week) until just beyond 13 months of age (late middle age in mice). Control mice, identically housed and maintained, received identical chow without the herb. Mice consuming untreated diet had a 79% survival by 10 months of age, while those consuming Echinacea daily in the diet were still 100% alive by 10 months. At approximately 13 months of age, mice consuming untreated diet had a 46% survival rate while those consuming Echinacea, were 74% alive at this time. Moreover, the key immune cells, acting as the first line of defense against developing neoplasms in mice and humans, i.e., natural killer (NK) cells, were significantly elevated in absolute number both in their bone marrow production site, as well as in the major organ to which they traffic and function, i.e., the spleen. The cells of the myeloid/granulocyte lineages remained steadfastly at control levels in both the bone marrow and spleen in Echinacea-consuming mice. Thus, it appears that regular intake of Echinacea may indeed be beneficial/prophylactic, if only for the reason that it maintains in an elevated state, NK cells, prime elements in immunosurveillance against spontaneous-developing tumors, a phenomenon which increases in frequency with progressive aging. PMID- 16041620 TI - Age-dependent increase of prolyl-4-hydroxylase domain (PHD) 3 expression in human and mouse heart. AB - The hypoxia-inducible factor (HIF)-1 is a master transcriptional activator of oxygen-regulated genes involved in energy metabolism, angiogenesis, and erythropoiesis. HIF-1 is composed of the two subunits HIF-1alpha and HIF-1beta (also called ARNT). The destruction of HIF-1alpha in the presence of oxygen is initiated by prolyl-4-hydroxylation. In human cells three closely related prolyl hydroxylases (PHDs) have been identified. An age-dependent decrease in HIF-1alpha expression was reported previously in brain, liver and kidney, which may be associated with a reduced adaptation to hypoxia as found in aged animals and humans. We have determined the expression of HIF-1alpha and the PHDs in human atrial trabeculae under normoxic and hypoxic conditions, in samples of human left ventricles as well as in heart extracts from female mice of different age (5 up to 16 months). With increasing age we found a decreased expression of HIF-1alpha, which correlated to an increased PHD3 expression in mouse and human heart. PHD3 was the most prominent HIF modifying hydroxylase found in human heart samples. Additionally, we found a strong ischemia/hypoxia-inducibility of PHD3 compared to PHD1 and PHD2 in atrial trabeculae. These data may explain the previously reported reduction of HIF-1alpha and HIF-1 target genes such as the vascular endothelial growth factor in ageing tissue. PMID- 16041621 TI - Regulation of p70S6k, GSK-3beta, and calcineurin in rat striated muscle during aging. AB - In this study we compared the content and phosphorylation levels of several molecules believed to regulate muscle hypertrophy and fiber type changes in the extensor digitorum longus (EDL), soleus, diaphragm, and heart of adult (6 months), aged (30 months), and very aged (36 months) Fischer 344 x Brown Norway rats. With aging, the mass of the EDL and soleus decreased significantly (approximately 38% and approximately 36%, respectively), the diaphragm's mass remained unchanged while the mass of the heart increased (approximately 35%). Western blotting demonstrated that calcineurin (CnA), the 70-kDa ribosomal S6 kinase (p70(S6k)), glycogen synthase kinase-3beta (GSK-3beta), and the phosphorylated forms of GSK-3beta and p70(S6k) (p-GSK-3beta(Ser9) and p p70(S6kThr389)) were regulated differently with aging and between muscle types. Total p70(S6k), GSK-3beta, and p-GSK-3beta(Ser9) decreased in the aged-atrophic EDL and soleus while p-p70(S6kThr389) increased. Although total p70(S6k) content diminished in the continuously active diaphragm, phosphorylation of p70(S6k )remained unchanged. Conversely, the expression of GSK-3beta and p-GSK 3beta(Ser9) increased in the diaphragm. With aging, the amount of p p70(S6kThr389) decreased approximately 56% in the heart while p-GSK-3beta( Ser9) increased approximately 193%. Interestingly, CnA content remained unchanged in the diaphragm, increased approximately 204% in the EDL, and decreased approximately 30% and approximately 65% with aging in the soleus and heart, respectively. These results indicate remarkable differences in the regulation of molecules thought to govern protein synthesis and changes in contractile protein expression. PMID- 16041622 TI - Age- and gender-related alterations of the number and clonogenic capacity of circulating CD34+ progenitor cells. AB - The aim of this study was to evaluate the peripheral representation and the clonogenic capacity of CD34(+) progenitor cells from 130 healthy subjects (80 females and 50 males) ranging in age from 16 to 100 years. We demonstrated that the absolute number of circulating CD34(+) cells progressively and significantly decreased with advancing age, with a 2-fold reduction in subjects aged more than 80 years. The number of granulocyte-macrophagic (CFU-GM), erytroid (BFU-E), and mixed (CFU-GEMM) colonies which developed from the number of CD34(+) purified cells per ml, progressively and significantly decreased with advancing age. The reduction of both CD34(+) cell number and clonogenic capacity during aging was statistically significant in males but not in females. When evaluated on a per cell bases, a significant age-related decrease in the number of CFU-GM colonies was observed in female but not in male subjects. Our study demonstrates the influence of gender on age-related alterations of the number and clonogenic capacity of CD34(+) cells in the peripheral blood. This evidence deserves particular consideration for the future planning of stem cell therapy in age associated debilitating diseases. PMID- 16041623 TI - Age-related susceptibility of naive and memory CD4 T cells to apoptosis induced by IL-2 deprivation or PHA addition. AB - The increased age-associated incidence of infectious and cancer diseases has been related to the alteration of immune functioning found in the elderly (immunosenescence). The reduction of naive T cells, which determine an impaired ability to mount immune responses to new antigens, represents a hallmark of the aging process. The aim of this study was to evaluate the susceptibility to apoptosis of purified naive and memory CD4(+) T cells from peripheral blood of healthy people ranging in age from 20 to 98 years. Two mechanisms of T cell elimination by apoptosis have been evaluated: cytokine deprivation and activation induced cell death. After Interleukin-2 deprivation, the percentage of naive and memory CD4(+) apoptotic cells significantly increased with donor age concomitantly with a reduction of Bcl-2 expression and an increase of intracellular content of reactive oxygen species. After phytohemagglutinin addition, the percentage of apoptotic cells, the expression of CD95, and the intracellular reactive oxygen species, were not significantly correlated with age both in naive and memory CD4(+) T cells. Our data demonstrate the existence of functional alterations of naive and memory T cell populations during ageing. These alterations are mainly related to the mechanism of the apoptotic event rather than to the type of cell population involved (naive or memory). The alterations of naive and memory T cells may have implications in the age-related susceptibility to diseases. PMID- 16041624 TI - Is aging the price for memory? AB - Aging (senescence) is apparent in animals that possess long-lived postmitotic cells but is negligible in primitive species, such as hydras and other Cnidarians, all of whose cells are constantly renewed by cell division. This repetitive mitotic activity precludes the progressive intracellular accumulation of damaged biomolecules and organelles, which are obvious concomitants of aging in neurons and other long-lived cells of higher animals. We assume that the development of long-lived postmitotic cells, now found in the overwhelming majority of species, represented a useful evolutionary change. Probably, of particular importance was the evolution of long-lived neurons, which are required for long-term memory. However, the appearance of long-lived postmitotic cells not only increased fitness, but also gave rise to the aging process. PMID- 16041625 TI - "My involvement in aging research was just a series of coincidences" An interview with Kenichi Kitani. Interview by Sataro Goto. PMID- 16041626 TI - Second IRCHAL conference and the communication of biogerontology to health care personnel. PMID- 16041627 TI - Immunohistochemical study of a membrane skeletal molecule, protein 4.1G, in mouse seminiferous tubules. AB - Protein 4.1 families have recently been established as potential organizers of an adherens system. In the adult mouse testis, protein 4.1G (4.1G) localized as a line pattern in both basal and adluminal compartments of the seminiferous tubules, attaching regions of germ cells and Sertoli cells. By double staining for 4.1G and F-actin, their localizations were shown to be different, indicating that 4.1G was localized in a region other than the basal and apical ectoplasmic specializations, which formed the Sertoli-Sertoli cell junction and Sertoli spermatid junction, respectively. By electron microscopy, immunoreactive products were seen exclusively on the cell membranes of Sertoli cells, attaching to the various differentiating germ cells. The immunolocalization of cadherin was identical to that of 4.1G, supporting the idea that 4.1G may be functionally interconnected with adhesion molecules. In an experimental mouse model of cadmium treatment, in which tight and adherens junctions of seminiferous tubules were disrupted, the 4.1G immunostaining in the seminiferous tubules was dramatically decreased. These results indicate that 4.1G may have a basic adhesive function between Sertoli cells and germ cells from the side of Sertoli cells. PMID- 16041628 TI - JNK/ERK-AP-1/Runx2 induction "paves the way" to cartilage load-ignited chondroblastic differentiation. AB - Chondro-osteogenesis and subsequently skeletal morphology are greatly influenced by mechanical loads. The exact mechanism(s) by which mechanical stimuli are transduced in chondrocytes remains obscure and appears to be equally complex with similar signal transducing systems. Here we investigated whether and to what extent the MAPK (JNK/ERK)-AP-1/Runx2 signaling pathways are engaged in this phenomenon, and assessed their involvement in the functional biology of articular cartilage. For this purpose, 14-day-old female Wistar rats were divided into 2 groups: the first group was fed hard diet (simulating physiologic temporomandibular joint (TMJ) loading), while the second group was fed soft diet (reduced TMJ loading). On day 21 (experiment initiation day - weaning day), biopsies from condyles of both groups were obtained after 6, 12 and 48 h of functional TMJ loading. Immunohistochemical methodology was employed to evaluate the expression levels of pc-Jun, c-Fos, JNK2, p-JNK, p-ERK and Runx2 due to alteration in functional load. Our data demsonstrate that the protein levels of all the aforementioned molecules were markedly increased in animals fed with the hard diet, throughout the experimental procedure. These results indicate that functional cartilage loading induces the AP-1 and Runx2 transcription factors through the JNK and ERK MAPK cascades. In as much as the above signaling mediators/effectors are considered to be crucial in the differentiation/maturation process of cartilage tissue, we pose that functional mechanical loading of condylar cartilage serves to "fine tune" chondroblastic differentiation/maturation. PMID- 16041629 TI - Randomized controlled trial of vaginal misoprostol versus vaginal misoprostol and isosorbide dinitrate for termination of pregnancy at 13-29 weeks. AB - When compared to the use of 12 hourly 200 microg vaginal misoprostol on its own, the addition of a single dose of 5 mg did not significantly increase the abortion or delivery rate in pregnancy termination at 13-29 weeks gestation. All patients were given intravenous syntocinon at 30 mU/min from the first dose of misoprostol onwards. PMID- 16041630 TI - Abnormal basement membrane in the inner ear and the kidney of the Mpv17-/- mouse strain: ultrastructural and immunohistochemical investigations. AB - The loss of the function of the peroxisomal Mpv17-protein and associated imbalanced radical oxygen species (ROS) homeostasis leads to an early onset of focal segmental glomerulosclerosis and sensorineural deafness associated with severe degeneration of cochlear structures. An excessive enlargement of basal laminae of the stria vascularis capillaries and glomeruli indicates numerous changes in their molecular composition. The basement membrane (BM) of the glomeruli and the stria vascularis are simultaneously affected in early stages of the disease and the lamination, splitting of the membrane and formation of the "basket weaving" seen at the onset of the disease in the kidney are similar to the ultrastructural alterations characteristic for Alporta9s syndrome. The progressive alteration of the BMs is accompanied by irregularity in the distribution of the collagen IV subunits and by an accumulation of the laminin B2(gamma1) in the inner ear and B(beta1) in the kidney. Since Mpv17 protein contributes to ROS homeostasis, further studies are necessary to elucidate downstream signaling molecules activated by ROS. These studies explain the cellular responses to missing Mpv17-protein, such as accumulation of the extracellular matrix, degeneration, and apoptosis in the inner ear. PMID- 16041631 TI - Lidocaine 4% cream compared with lidocaine 2.5% and prilocaine 2.5% or dorsal penile block for circumcision. AB - This study evaluated the efficacy and safety of lidocaine 4% cream (LMX4), compared with lidocaine 2.5% and prilocaine 2.5% (EMLA) or dorsal penile block (DPNB) for analgesia during circumcision. Healthy, term males (n = 54), younger than 1 week old undergoing circumcision were randomly assigned to open-label pretreatment with LMX4, EMLA, or DPNB. Heart rate (HR; beats per minute [bpm]), respiratory rate (RR; breaths/minute), and arterial oxygen saturation as measured by pulse oximetry (Sp O2; %) were monitored at baseline, and during drug application, circumcision, and recovery. Mean differences were compared using the general linear model. At the end of drug application, mean HR for infants receiving LMX4 (146 bpm; standard error of mean [SEM], 8.0 bpm) was lower than that for DPNB (176 bpm; SEM, 8.3 bpm; p < 0.05). No significant difference in mean HR was observed between treatments during circumcision. Mean RR was higher during circumcision for EMLA compared with LMX4 (p < 0.05) and DPNB (p < 0.05). At lysis, mean RR was significantly lower in DPNB than LMX4 and EMLA. The number of Sp O2 samples was too small for comparison. Three infants (one receiving LMX4 and two receiving EMLA) experienced local reactions (p = 0.54). No adverse effects were observed with DPNB. No difference in analgesic efficacy was observed between treatments according to HR. Differences in RR may reflect a varying level of analgesia. The safety profile was similar for all treatments. LMX4 is an effective analgesic for newborn circumcision. PMID- 16041632 TI - Neonatal ergot poisoning: a persistent iatrogenic illness. AB - Ergot toxicity in the newborn usually manifests itself as respiratory depression, cyanosis, oliguria, and seizures. Death is usually caused by respiratory failure. A limited number of neonatal cases have been reported worldwide, and almost all cases involved confusion of maternal methylergonovine with neonatal vitamin K. Previous case reports provided little information regarding the effectiveness and dosing of antidotal therapy, especially sodium nitroprusside. A full-term male infant was inadvertently given methylergonovine instead of naloxone at birth. Several hours later, he required intubation for respiratory failure. Peripheral perfusion, ventilation, and renal function improved rapidly with nitroprusside infusion, and he was extubated on the third hospital day. Even asymptomatic newborns should be transferred to a neonatal intensive care unit for close observation after methylergonovine administration because toxicity can be life threatening. Rapid recognition of the therapeutic error, ventilatory support, and prompt administration of sodium nitroprusside should lead to a good outcome. PMID- 16041633 TI - The influence of race on fetal outcome. AB - The purpose of this study was to examine racial differences in fetal death by gestational age among South Carolinians during 1999 to 2000 in an attempt to identify sociodemographic risk factors that may contribute to the refinement of care protocols. This historical cohort design appended the 1999 to 2000 South Carolina Vital Records fetal death file (N = 944) to the birth file (N = 99, 726) to perform secondary data analysis. Inclusion criteria were maternal South Carolina residency, delivery of a singleton in South Carolina, and racial identification as white or black. Independent associations between race (black or white), gestational age categories of deliveries (fetal death or live birth), and maternal sociodemographic characteristics (maternal age and prenatal care) were examined using chi2 analysis. Black and white fetal mortality rates (FMRs) were examined. Logistic regression was used to control for confounding variables. The study sample consisted of 63.4% white and 36.6% black for all deliveries; 40.89% white and 59.11% black for fetal deaths. There was a significant racial difference in gestational age categories, initiation of prenatal care, maternal age, and fetal death rates (p = 0.0001). The black FMR was nearly 2.5 times greater than the white rate (p < 0.0001; 95% confidence interval, 2.2 to 2.9). There was an independent association between race and gestational age at fetal death (p = 0.0001) as well as race and maternal age. Results did not remain statistically significant after controlling for confounding variables. In this study, we identified several factors associated with an increased risk of fetal death. However, after controlling for gestational age, the odds of fetal death among blacks were no longer significantly elevated in comparison to those of whites. Future studies that examine racial disparities should consider gestational age in analytic models. Identifying racial demographics and behavioral risks may contribute to refinements in care protocols for high-risk mothers in an effort to reduce fetal mortality. PMID- 16041634 TI - Timing of umbilical cord separation and neonatal intensive care unit practices. AB - We investigated the effects of antibiotic and corticosteroid treatment of the mother close to birth and postnatal neonatal intensive care unit (NICU) practices such as phototherapy, parenteral nutrition, and antibiotic treatment of the infant on umbilical cord detachment time. The impact of dry cord care on risk of omphalitis in NICU patients was also investigated. Cord separation time was longer in preterm babies. Antenatal antibiotic and corticosteroid use did not affect cord detachment time. The use of postpartum antibiotics, parenteral nutrition, and phototherapy delayed cord separation. Among these, antibiotic treatment was the most effective factor. Dry cord care did not increase the risk of omphalitis in NICU patients in our series. PMID- 16041635 TI - Sterile water gastric drip in extremely low birthweight premature infants: a randomized trial. AB - This study was performed to test the hypothesis that sterile water gavage drip (SWGD) used in the fluid management of extremely low birthweight (ELBW) infants will decrease the incidence of hypernatremia. Secondary hypotheses included decreased hyperkalemia, hyperglycemia, and hyperbilirubinemia. Sixty ELBW infants were randomized before 36 hours of age to receive SWGD (up to 30 mL/kg/d) and intravenous fluid or conventional intravenous fluid management. SWGD was well tolerated in 89% of the infants. No difference was seen in the incidence of hypernatremia, hyperkalemia, hyperglycemia, or hyperbilirubinemia. A significant reduction in the incidence of treated patent ductus arteriosus (PDA) was noted in the study group (36% versus 69%; relative risk, 0.52; 95% confidence interval, 0.30 to 0.90; p = 0.02). SWGD may provide an alternative means of safely administering free water to the ELBW infant. The observed reduction in treated PDA requires further investigation. PMID- 16041636 TI - Meconium mineral content in small for gestational age neonates. AB - The mineral concentration of meconia of small for gestational age (SGA) newborns were compared with those of appropriate for gestational age (AGA) newborns of similar gestational ages (GA) to determine whether differences may provide clues of possible nutritional deficits of SGA infants, given that levels of meconium minerals could indicate the use of minerals by the fetus and the sufficiency of the maternal supply of minerals. Twenty-one SGA and 24 AGA newborns were included. Eleven SGA and 15 AGA were < or = 35 weeks GA. Ten SGA and nine AGA infants were > or = 36 weeks GA. All meconia from each neonate was processed and assayed for iron, zinc, copper, manganese, calcium, magnesium, and phosphorus. In the < or = 35-week subgroups, the SGA infants had lower meconium iron and manganese concentrations than that of the AGA. Among > or = 36-week newborns, SGA infants had a higher birthweight-adjusted copper concentration than AGA infants, but no differences were observed for the remaining elements. Lower iron and manganese meconium in < or = 35-week SGA infants may reflect either a greater use or a decreased maternal supply. The higher birthweight-adjusted meconium copper in the > or = 36-week SGA infants may be due to a comparatively reduced fetal use or increased maternal supply. These data may assist in clarifying potential mechanisms affecting intrauterine growth and/or potential nutrient deficits in the neonatal period. PMID- 16041637 TI - Ultrasound screening for congenital pyelectasis in term pregnancies. AB - The purpose of this study was to compare the role of antenatal ultrasound screening for congenital pyelectasis at term with postnatal screening. An ultrasonographic prospective study of 231 fetuses of low-risk pregnancies was performed, evaluating the renal pelvis at term (38 to 42 weeks gestation) and after birth (first and third months of life). Cut-off points used were 9 mm antenatally, and 10 and 12 mm, respectively postnatally, all in an anteroposterior view. A sensitivity of 20% and a specificity of 93.4% was obtained when comparing the antenatal versus the third-month scan, whereas when we compared the sonograms from the first versus the third month of life, we found a sensitivity of 60% and a specificity of 89.7%, with a positive predictive value of 14.7% in the first case and 25% in the second case. A p value < 0.001 was obtained when comparing the sensitivities; a p value of 0.2 was reported when comparing the specificities. Data on a comparative screening of the renal pelvis measured antenatally at term and postnatally do not exist. Although it is extremely feasible, the measurement of the fetal renal pelvis during the routine ultrasound scan at term of gestation seems of no benefit in terms of sensitivity and positive predictive values in identifying infants affected by congenital pyelectasis. Because of the low values of sensitivity obtained in our study, a screening program for the renal pelvis dilation both in fetuses and newborns cannot be recommended. PMID- 16041638 TI - Perinatal outcomes in growth-restricted twins compared with age-matched growth restricted singletons. AB - The objective of this study was to compare the perinatal outcomes in growth restricted (IUGR) twins with a matched group of IUGR singletons. We performed a retrospective cohort study evaluating the perinatal outcomes in IUGR twins matched for gestational age at delivery to the next four IUGR singletons (1:4) in our perinatal database. The perinatal outcomes evaluated included rates of uncorrected perinatal mortality (PMR), admission to the neonatal intensive care unit (NICU), respiratory distress syndrome, grades 3 or 4 intraventricular hemorrhage, necrotizing enterocolitis, periventricular leukomalacia, and length of NICU stay. A cohort of twins with IUGR were matched with a cohort of IUGR singletons and compared for perinatal outcomes. A p value < 0.05 was considered significant. During the study period, we identified 99 IUGR twins meeting our inclusion criteria. These were matched with 396 singletons with IUGR. The mean gestational age at delivery for the twins was 34 +/- 3.8 weeks and 35 +/- 3.5 weeks for the singletons ( p = 0.11). The PMR in twins was significantly higher than in singletons (OR, 2.2; 95% confidence interval, 1.1 to 5.7). Perinatal morbidity was similarly significantly higher in IUGR twins. Compared with age matched singletons, twins with IUGR have higher perinatal mortality and morbidity rates. PMID- 16041639 TI - Atrial flutter following placement of an umbilical venous catheter. AB - Atrial flutter is a rare complication of the insertion of an umbilical venous catheter. We report a neonate who had an umbilical venous catheter placed within the first 24 hours of life and developed a hemodynamically stable tachyarrhythmia. The arrhythmia was successfully managed with synchronized cardioversion, with no further complications. The presentation and management are discussed. Physicians should be watchful for such potentially life-threatening complications from the placement of indwelling lines. PMID- 16041640 TI - Fetal and neonatal mortality among twin gestations in a Canadian population: the effect of intrapair birthweight discordance. AB - The purpose of this study was to examine the association between intrapair birthweight discordance and fetal and neonatal mortality. This was a historical cohort study, using the Statistics Canada linked birth-infant death file. Twin births in Canada (excluding Ontario) for the period 1986-1997 were studied. Outcome measures were fetal and neonatal mortality. Among 59,034 twin births, 53% had 0 to 9% birthweight difference; 30% had 10 to 19% discordance; 11% had 20 to 29% discordance; and 6% had > or = 30% discordance. Maternal age, parity, sex discordance, and gestational age were important factors affecting birth weight discordance. Fetal death rates were 0.95, 1.26, 3.14, and 11.10%, respectively, in infants with a birthweight discordance of 0 to 9, 10 to 19, 20 to 29, and > or = 30%. Corresponding figures for neonatal death rates were 1.90, 2.32, 3.05, and 8.86%, respectively. Adjustment for important confounding variables including the actual birthweight and gestational age substantially reduced the birthweight discordance-related risk of fetal and neonatal mortality, but the results remained statistically significant and clinically important. The birthweight discordance-related risk of fetal and neonatal mortality was higher in smaller twins than in larger twins and higher in infants with gestational age > or = 37 weeks than those < 37 weeks. Birthweight discordance is an important risk factor for fetal and neonatal mortality, and the birthweight discordance-related risk of fetal and neonatal mortality is higher in smaller twins than in larger twins and higher in term twins than in preterm twins. PMID- 16041641 TI - The Neuroprotection of puerarin against cerebral ischemia is associated with the prevention of apoptosis in rats. AB - Previous work has shown that puerarin (Pur), extracted from the dried root of Pueraria lobata (Wild) Ohwi, increases cerebral blood flow in dogs and attenuates cerebral and spinal cord injury resulting from ischemia and reperfusion in rats and rabbits. The present study further demonstrates the neuroprotective effects of Pur on cerebral ischemic injury in rats and the mechanisms underlying the protective effects. Male Sprague-Dawley rats were subjected to middle cerebral artery occlusion (MCAo) for 50 min followed by reperfusion for 24 h. Pur (50, 100 mg/kg, i.p) was administered at the onset of MCAo. Twenty-four hours after reperfusion, neurological deficits were evaluated in Pur- and vehicle-treated rats. The infarct volume and edema ratios were assessed from stained brain slices. The results showed that Pur (100 mg/kg) markedly decreased the infarct volume by 34 % ( P < 0.01) in cerebral cortex and improved the neurological functions ( P < 0.05) after MCAo. Furthermore, flow cytometric analysis of annexin-V and PI labeling cells showed that the percentages of apoptosis and necrosis in the dorsolateral cortex were significantly reduced by 38.6 % and 28.5 % ( P < 0.01 and P < 0.05) following treatment with Pur (100 mg/kg) in MCAo rats. Caspase-3 activity, a biochemical marker of apoptosis, was significantly inhibited after treatment with Pur in the dorsolateral cortex. In agreement with this result, the expression of the X-chromosome-linked inhibitor of apoptosis protein (XIAP) was obviously up-regulated after administration of Pur (100 mg/kg), while caspase-3 gene was down-regulated in the dorsolateral cortex. These results suggest that the neuroprotection of puerarin against cerebral ischemia is associated with anti-apoptosis. PMID- 16041642 TI - Preserved pharmacological activity of hepatocytes-treated extracts of valerian and St. John's wort. AB - The two herbal extracts valerian (Valeriana officinalis L.) and St. John's wort (Hypericum perforatum L.) were studied for their metabolic changes upon incubation with freshly prepared rat hepatocytes and subsequently analysed phytochemically as well as pharmacologically in vitro. Quantitative HPLC analysis of valerian extracts revealed considerable metabolic activities with regard to sesquiterpenes and iridoids. The amount of acetoxyvalerenic acid decreased 9 fold, while that of hydroxyvalerenic acid correspondingly increased 9-fold due to O-deacetylation. The valepotriates didrovaltrate, isovaltrate and valtrate decreased 2-, 18- and 16-fold, respectively. However, the binding affinities of the incubated extracts to the benzodiazepine and picrotoxin binding site of the GABA (A) receptor were quite similar to those of the non-incubated extracts. Neither valerenic acids nor valepotriates exhibited any significant effect on the two binding sites when tested as single compounds. Therefore, either other constituents represent the active ones or multiple compounds are necessary for the observed inhibitory and allosteric effects at the GABA (A) receptor. Extracts of St. John's wort were less potently metabolised than valerian. The amount of pseudohypericin and the main flavonoids (hyperoside, rutin, isoquercitrin, quercitrin, quercetin and I3,II8-biapigenin) slightly decreased during the 4-h incubation period. Both the antagonist effect at the corticotropin-releasing factor (CRF) type 1 receptor and the binding inhibition at the 5-HT transporter were attenuated during the metabolic treatment. The reduced antagonist effect correlates with the decreasing amount of pseudohypericin known to be a CRF (1) receptor antagonist. In conclusion, the incubation of plant extracts with freshly prepared rat hepatocytes represents a useful approach to study the pharmacological action of metabolised plant extracts. The consistent pharmacological activity of both valerian and St. John's wort is concordant with the known clinical efficacy of pharmacological activities. PMID- 16041643 TI - Inhibition of PDGF beta-receptor tyrosine phosphorylation and its downstream intracellular signal transduction in rat aortic vascular smooth muscle cells by kaempferol. AB - Kaempferol, a flavonoid present in human diet and plants, has been known to show cardiovascular protection via its anti-oxidant activity. In this study, we have investigated the effect of kaempferol on the proliferation of primary cultured rat aortic vascular smooth muscle cells (VSMCs). Kaempferol significantly inhibited 50 ng/mL platelet-derived growth factor (PDGF)-BB-induced proliferation and [3H]-thymidine incorporation into DNA at concentrations of 5, 20 and 50 microM without any cytotoxicity. Kaempferol also inhibited the c-fos mRNA expression induced by PDGF-BB concentration-dependently. In addition, consistent with the inhibition of cell proliferation and c-fos mRNA expression, kaempferol inhibited the PDGF beta-receptor (Rbeta) phosphorylation in a concentration dependent manner. Accordingly, the downstream signal transductions of PDGF-Rbeta such as ERK1/2, Akt and PLC-gamma1 phosphorylation were also inhibited by kaempferol in the same pattern. These findings suggest that, in addition to its anti-oxidant activity, the cardiovascular protective effect of kaempferol may be mediated, at least in part, by the suppression of VSMC proliferation, which is due to the inhibition of PDGF-Rbeta tyrosine phosphorylation and its downstream intracellular signal transduction. PMID- 16041644 TI - 14-deoxyandrographolide as a platelet activating factor antagonist in bovine neutrophils. AB - 14-Deoxyandrographolide (14-DAP) is a labdane diterpene isolated from Andrographis paniculata with previously reported calcium channel blocking activity. Its potential platelet activating factor (PAF) antagonistic activity in bovine neutrophils was assessed. 14-DAP, in concentrations between 10-100 microM, reduced the extracellular acidification rate and the intracellular alkalinization in a dose-dependent manner. In addition, 14-DAP reduced PAF-induced calcium flux in the presence of extracellular calcium, and tyrosine phosphorylation of a 44 kDa protein corresponding to the MAPK(ERK1). However, 14-DAP reduced the 3H-PAF binding with a Ki of 7.8 x 10 (- 9)M, and a Hill slope of 0.63, suggesting that there is more than one binding site for 14-DAP. We concluded that 14-DAP is an effective antagonist of PAF-mediated processes in bovine neutrophils, probably by virtue of its calcium channel blocking property. PMID- 16041645 TI - 5-HT3 receptor blocking activity of arylalkanes isolated from the rhizome of Zingiber officinale. AB - Different extracts (ethanolic, hexane, aqueous) of ginger (rhizomes of Zingiber officinale) and the essential oil were tested using [14C]guanidinium influx into N1E-115 cells and the isolated rat ileum in order to identify their activity in inhibiting 5-HT3 receptor function. The hexane extract proved to be the most active and yielded upon bioassay-guided fractionation nine constituents: [6]-, [8]-, [10]-gingerols, [6]- and [8]-shogaols which were previously shown as active in vivo against cytotoxic drug-induced emesis; [4]-gingerol, [6]-gingerdiol, diacetyl-[6]-gingerdiol and [6]-dehydrogingerdione have not been previously tested for anti-emetic or 5-HT3 receptor antagonistic effects. Even though the latter four compounds are only minor constituents, their identification contributed towards the characterisation of a structure-activity relationship of this class of compounds. The order of potency for the nine constituents in the N1E-115 cell system was [6]-gingerdiol approximately diacetyl-[6]-gingerdiol approximately [6]-dehydrogingerdione approximately [6]-shogaol > or = [8]-shogaol approximately [8]-gingerol > [10]-gingerol > or = [6]-gingerol > [4]-gingerol. PMID- 16041646 TI - Myricetin as the active principle of Abelmoschus moschatus to lower plasma glucose in streptozotocin-induced diabetic rats. AB - The antihyperglycemic action of myricetin, purified from the aerial part of Abelmoschus moschatus (Malvaceae), was investigated in streptozotocin-induced diabetic rats (STZ-diabetic rats). Bolus intravenous injection of myricetin decreased the plasma glucose concentrations in a dose-dependent manner in STZ diabetic rats. Myricetin at the effective dose (1.0 mg/kg) significantly attenuated the increase of plasma glucose induced by an intravenous glucose challenge test in normal rats. A stimulatory effect of myricetin on glucose uptake of the soleus muscles isolated from STZ-diabetic rats was obtained in a concentration-dependent manner from 0.01 to 10.0 micromol/L. The increase of glucose utilization by myricetin was further characterized using the enhancement of glycogen synthesis in isolated hepatocytes of STZ-diabetic rats. These results suggest that myricetin has an ability to enhance glucose utilization to lower plasma glucose in diabetic rats lacking insulin. PMID- 16041647 TI - In vivo biodistribution of ginkgolide B, a constituent of Ginkgo biloba, visualized by MicroPET. AB - The in vivo dynamic behavior of ginkgolide B (GB), a terpene lactone constituent of the Ginkgo biloba extracts, in the living animal was visualized by positron emission tomographic (PET) imaging using a GB analogue labeled with the positron emitter (18)F. The in vivo imaging studies, combined with ex vivo dissection experiments, reveal that GB exists in 2 forms in the body: the original GB with its lactone rings closed and a second form with one of the rings open. The original GB in plasma is taken up rapidly by various organs including the liver, the intestine and possibly the stomach. Consequently, in plasma, the proportion of the ionized form of GB increases dramatically with time. Thereafter the ratio between the 2 forms appears to shift slowly towards equilibrium. The results suggest that more attention needs to be focused on in vivo dynamics between the 2 forms of GB. PMID- 16041648 TI - Fucan inhibits Chinese hamster ovary cell (CHO) adhesion to fibronectin by binding to the extracellular matrix. AB - In recent years, sulfated fucans have emerged as an important class of natural biopolymers. In this study, the anti-adhesive activity of a fucan from the brown seaweed Spatoglossum schroederi was analyzed using tumorigenic cells: wild-type Chinese hamster ovary cells (CHO-K1) and the mutant type deficient in xylosyltransferase (CHO-745). Fibronectin (FN) was used as substrate for cell attachment. For both cell types, this fucan has shown a dose-dependent anti adhesive effect, reaching saturation at around 400 mug/mL. This effect was abolished by desulfation of the fucan. In addition, this polymer exhibited the highest inhibitory effect in comparison to other sulfated polysaccharides. The fucan was biotinylated and used as a probe to identify its action sites. Biotinylated fucan was detected in the extracellular matrix environment by confocal microscopy and flow cytometric analysis, but not at the cell surface. The results suggest that the fucan shows anti-adhesive activity by binding directly to FN, and blocking FN sites that are recognized by cell surface ligands, possibly the integrin family. PMID- 16041649 TI - Carvacrol and eugenol differentially stimulate intracellular Ca2+ mobilization and mitogen-activated protein kinases in Jurkat T-cells and monocytic THP-1 cells. AB - Essential oils are a major active constituent found in many Chinese medicinal herbs. Here, we demonstrate that two components of essential oils, carvacrol and eugenol, dose-dependently trigger intracellular Ca2+ mobilization in Jurkat T cells and THP-1 monocytic cells. Both carvacrol and eugenol are also capable of stimulating the active phosphorylation of the p38 subgroup of mitogen-activated protein kinases (MAPKs) in both cell types. However, carvacrol selectively activated the ERK subgroup in Jurkat T-cells, and stimulated the JNK subgroup in THP-1 monocytic cells. Eugenol treatment was not linked to ERK or JNK activation in either cell type. EC50 values for the induction of Ca2+ mobilization and MAPK activation were around 10 - 30 microM for both carvacrol and eugenol. Our results suggest that these essential oil components may act as effective agents to modulate the functions of immuno-responsive cells via different intracellular signaling pathways. PMID- 16041650 TI - Effects of Evodia rutaecarpa and rutaecarpine on the pharmacokinetics of caffeine in rats. AB - To assess possible herb-drug interactions, rutaecarpine (an herbal ingredient of Evodia rutaecarpa; 25 mg/kg/day, p. o.), the ethanol extract of Evodia rutaecarpa (1 g/kg/day, p. o.), and an herbal preparation of Evodia rutaecarpa (Wu-Chu-Yu Tang; 1 g/kg/day) were individually pretreated daily for three consecutive days in rats and on the fourth day caffeine was administered (2 mg/kg, i. v.). Caffeine concentrations in blood, brain and bile were concurrently measured by microdialysis coupled to a liquid chromatographic system. Pharmacokinetic data were calculated by a non-compartmental model. The results indicate that the caffeine crosses the blood-brain barrier and goes through hepatobiliary excretion. The caffeine level was significantly decreased by the pretreatment of rutaecarpine, the extract of Evodia rutaecarpa and herbal preparation Wu-Chu-Yu Tang. This finding should be very important whenever herb-drug interactions would be possible for a herbal remedy. PMID- 16041651 TI - Antihepatitis activity (anti-HBsAg and anti-HBeAg) of C19 homolignans and six novel C18 dibenzocyclooctadiene lignans from Kadsura japonica. AB - Bioassay-directed fractionation of the EtOAc extract of Kadsura japonica has led to the isolation of six new C18 dibenzocyclooctadiene lignans, schizanrins I, J, K, L, M, N, along with four known C19 homolignans, taiwanschirins A, B, C, and heteroclitin F. The elucidations of the new structures were based on spectral analysis. Bioassay evaluation against human type B hepatitis revealed that taiwanschirins A and B showed strong activity for anti-HBsAg and a medium effect for anti-HBeAg at 25 microg/mL (12.9 and 11.9 microM for taiwanschirins A and B, respectively). PMID- 16041652 TI - Brosimacutins J-M, four new flavonoids from Brosimum acutifolium and their cytotoxic activity. AB - Four new flavonoids, brosimacutins J-M (1 - 4), were isolated from the bark of Brosimum acutifolium Huber together with a known flavan, brosimine A (5). The structures of compounds 1-4 were elucidated by spectroscopic means. 27 constituents of this plant including compounds 1-5 were evaluated for their cytotoxic activity against murine leukemia P388 cells. Although no compounds tested had any reversal effect on vincristine resistance, brocimacutins J-M (1-4) were cytotoxic to vincristine-resistant P388 cells (IC50 4.4 - 19 microg/mL). PMID- 16041653 TI - New cytotoxic biflavonoids from Selaginella delicatula. AB - Five new biflavonoids, robustaflavone 7,4',4'''-trimethyl ether, robustaflavone 4',4'''-dimethyl ether, 2,3-dihydroamentoflavone 7,4',7''-trimethyl ether, 2,3 dihydroamentoflavone 7,4'-dimethyl ether, and 2'',3''-dihydroisocryptomerin 7 methyl ether, together with six known compounds have been isolated from the aerial parts of Selaginella delicatula. The structures of these new compounds were determined through spectral analyses. Among the isolates, robustaflavone 4',4'''-dimethyl ether, 2,3-dihydroamentoflavone 7,4'-dimethyl ether, and alpha tocopheryl quinone exhibited cytotoxicities (ED50 values < 4 microg/mL) against P 388 and/or HT-29 cell lines in vitro. PMID- 16041654 TI - Five new cytotoxic triterpenoid saponins from the roots of Symplocos chinensis. AB - Five new triterpenoid saponins, named symplocososides G-K, were isolated from the roots of Symplocos chinensis. Their structures were elucidated by spectral and chemical methods as symplocososide G, 3beta-O-{[beta- D-glucopyranosyl(1- >2)][alpha-L-arabinofuranosyl(1-->4)]-beta-D-(3-O-acetyl)-glucuronopyranosyl} 21beta- O-[(2 Z)-3,7-dimethyl-2,6-octadienoyl]-22 alpha-O-(2-methylbutanoyl)-R1 barrigenol, symplocososide H, 3beta-O-{[beta-D-glucopyranosyl(1-->2)][alpha-L arabinofuranosyl(1-->4)]- beta-D-(3-O-acetyl)-glucuronopyranosyl}-21beta-O [(2E)3,7-dimethyl-2,6-octadienoyl]-22alpha-O-(2-methylbutanoyl)-R1-barrigenol, symplocososide I, 3beta-O-{[beta-D-glucopyranosyl(1-->2)][ alpha-L arabinofuranosyl(1-->4)]-beta-D-(3- O-acetyl-6-O-methyl)-glucuronopyranosyl} 21beta-O-[(2 Z)3,7-dimethyl-2,6-octadienoyl]-22alpha-O-(2-methylbutanoyl)-R1 barrigenol, symplocososide J, 3 beta-O-{[ beta-D-glucopyranosyl(1-->2)][alpha-L arabinofuranosyl(1-->4)]-beta-D-(3- O-acetyl)-glucuronopyranosyl}-21beta-O-[(2 Z)3,7-dimethyl-2,6-octadienoyl]-22alpha-O-benzoyl-R1-barrigenol, and symplocososide K, 3beta-O-{[beta-D-glucopyranosyl (1-->2)][alpha-L arabinofuranosyl(1-->4)]- beta-D-(3-O-acetyl-6-O-methyl)-glucuronopyranosyl} 21beta-O-[(2Z)3,7-dimethyl-2,6-octadienoyl]-22alpha-O-benzoyl-R1-barrigenol. Symplocososides G-K showed significant cytotoxicity against cancer cell lines KB, HCT-8, Bel-7402, BGC-823 and A549 with IC50 values ranging from 0.82 microM to 5.09 microM, except for symplocososide I against cancer cell lines KB, BGC-823, A549 and symplocososide K against cancer cell line BGC-823 with IC50 values >10.00 microM. PMID- 16041655 TI - Five new oleanolic acid glycosides from Achyranthes bidentata with inhibitory activity on osteoclast formation. AB - Bioassay-directed fractionation of a butanol-soluble fraction of methanol extract of the root of Achyranthes bidentata resulted in the isolation of 5 new oleanolic acid glycosides 1-5, namely, 18-(beta-D-glucopyranosyloxy)-28-oxoolean-12-en 3beta-yl 3-O-(beta-D-glucopyranosyl)-beta-D-glucopyranosiduronic acid methyl ester (1), achyranthoside C dimethyl ester (2), achyranthoside C butyl dimethyl ester (3), achyranthoside E dimethyl ester (4), and achyranthoside E butyl methyl ester (5), together with 10 known compounds. Their structures were established on the basis of spectroscopic interpretation and chemical methods. All the oleanolic acid glycosides inhibited the formation of osteoclast-like multinucleated cells (OCLs) induced by 1alpha,25(OH)2D3 in a co-culture assay system. PMID- 16041656 TI - Cytotoxicities of xanthones and cinnamate esters from Hypericum hookerianum. AB - 5-Hydroxy-2-methoxyxanthone (1), 2-hydroxy-3-methoxyxanthone (2), trans-kielcorin (3), 4-hydroxy-3-methoxyphenyl ferulate (4) and 3beta-O-caffeoylbetulinic acid (5) were isolated from Hypericum hookerianum. Compounds 1-5 were tested against the growth of three human tumor cell lines, MCF-7, NCI-H460 and SF-268. Compounds 4 and 5 exhibited significant inhibitory activity effects against all three; GI50 values for 4 were 15.1 +/- 1.6, 18.7 +/- 2.3 and 15.9 +/- 2.7 and for 5 12.2 +/- 2.4, 19.6 +/- 2.3 and 24.3 +/- 2.5. Compound 3 was less active with GI50 values of 55.1 +/- 2.3, 49.7 +/- 3.0 and 40.5 +/- 1.5, while 1 and 2 exhibited only weak effects. Compounds 4 and 5 were moderately effective in influencing the mitogenic response to human lymphocytes to hemoagglutinin, with IC values of 26.1 +/- 3.6 and 40.8 +/- 4.9, respectively. PMID- 16041657 TI - Antimitotic properties of pterocarpans isolated from Platymiscium floribundum on sea urchin eggs. AB - This study reports the antimitotic effects on sea urchin eggs of five known pterocarpans: (+)-3,10-dihydroxy-9-methoxypterocarpan, (+)-3,9 dimethoxypterocarpan [(+)-homopterocarpin], (+)-2,3,9-trimethoxypterocarpan, (+) 3,4-dihydroxy-9-methoxypterocarpan [(+)-vesticarpan] and (+)-3-hydroxy-9 methoxypterocarpan [(+)-medicarpin], isolated from the trunk of Platymiscium floribundum, a native tree from Brazil. All tested compounds showed strong activity in this assay, with 2,3,9-trimethoxypterocarpan being the most active (log IC50 of -8.10 +/- 0.02; -7.91 +/- 0.01; -7.97 +/- 0.02 M for first and third cleavages, and blastulae stages, respectively). These data suggest that the 2 methoxy substituent can be an important pharmacophoric unit. PMID- 16041658 TI - Isolation of cupanioside, a novel cytotoxic and antibacterial long-chain fatty alcohol glycoside from the bark of Cupania glabra. AB - The crude dichloromethane extract from the stem bark of Cupania glabra (Sapindaceae), showed in vitro cytotoxic activity against Hep G2, MDA-MB-231, Hs 578T, MCF-7, and PC-3 cells, and antibacterial activity against Bacillus cereus, Staphylococcus aureus, and Escherichia coli. Bioactivity-directed fractionation led to isolation of the new 1-O-[2'',3'',4''-tri-O-acetyl-alpha-L-rhamnopyranosyl (1-->2)-beta- D-glucopyranosyl]-hexadecanol (cupanioside) as the cytotoxic agent. The structure was elucidated by analysis of 2D-NMR spectra. PMID- 16041660 TI - Immune hemolytic anemia-a primer. PMID- 16041661 TI - Autoimmune pathogenesis and autoimmune hemolytic anemia. AB - Autoimmune hemolytic anemia (AIHA) is an autoimmune disorder in which autoantibodies are directed against an individual's own red blood cells (RBCs), leading to enhanced clearance through Fc receptor (FcR)-mediated phagocytosis. Although there is a large literature relating to clinical aspects of AIHA, relatively little work addresses how IgG autoantibodies are actually produced against RBC autoantigens. This review will first discuss the current understanding of autoimmunity in general and then focus on the knowledge of the immunopathogenic mechanisms responsible for autoantibody production in AIHA. Both human and animal studies will be discussed. Understanding theses mechanism is vital for developing antigen-specific immunotherapies to treat the disease. PMID- 16041662 TI - Treatment of autoimmune hemolytic anemia. AB - The appropriate therapy of autoimmune hemolytic anemia (AIHA) is dependent on the correct diagnosis and classification of this family of hemolytic disorders. Although the majority of cases are warm AIHA, there are several distinct types of cold AIHA and a number of drug-induced etiologies of AIHA, which must be investigated to determine if stopping a drug will induce a remission. In warm AIHA, corticosteroids are standard, followed by consideration of splenectomy in recalcitrant cases. If steroids and splenectomy are insufficient, other forms of immunosuppressive therapy are typically initiated. In cold AIHA, keeping the patient warm in often sufficient, but therapy directed at an underlying lympholiferative disorder may be helpful. Brisk hemolysis, inadequate responses to therapy, and worsening anemia require transfusion therapy. Although the pretransfusion workup is made difficult by the presence of the autoantibody, transfusion services can usually provide blood safe for transfusion by excluding underlying alloantibodies. When transfusion is urgently required and compatible blood cannot be located, incompatible blood may be provided as a life-saving measure. Communication between the transfusion service and the hematologist is critical to assess the risks in these settings. Hemoglobin-based oxygen carriers may provide an important bridging therapy in the future. Requests for "least incompatible" blood do not enhance transfusion safety and often result in unnecessary delays. PMID- 16041663 TI - The changing spectrum of drug-induced immune hemolytic anemia. AB - Drug-induced immune hemolytic anemia (DIIHA) occurs rarely. To date, about 100 drugs have been implicated in causing DIIHA and/or a positive direct antiglobulin test (DAT). The most common drugs associated with DIIHA in the 1970s were methyldopa and penicillin; currently, they are cefotetan and ceftriaxone. Drug antibodies fall into two types: drug-independent ("autoantibodies") and drug dependent ("penicillin type" or "immune complex type"); some patients have combinations of these antibodies. Some drugs cause nonimmunologic protein adsorption onto drug-treated red blood cells (RBCs). This is known to be the cause of positive indirect antiglobulin tests and is suspected to be a cause of positive DATs. This mechanism may be associated with hemolytic anemia. Twelve cephalosporins have been reported to cause DIIHA; five (primarily cefotetan and ceftriaxone) have been associated with fatalites. Patients with DIIHA due to cefotetan may only have received one dose of the drug prophylactically with surgery. Antibodies to cefotetan react to very high titers against drug-treated RBCs (and at lower titers against untreated RBCs without and/or with drug present). Patients with ceftriaxone-induced DIIHA have received the drug previously; reactions in children often occur minutes after ceftriaxone administration. Antibodies to ceftriaxone are only of the "immune complex type." PMID- 16041664 TI - Immune hemolysis associated with transplantation. AB - Immune hemolysis is one of the adverse effects that can occur following hematopoietic cell or solid organ transplantation. Understanding the clinical settings and the various causes of immune hemolysis is necessary for prompt diagnosis and appropriate management. One of the important causes is the passenger lymphocyte syndrome, which occurs following minor ABO blood group incompatibility between donor and recipient. Hemolysis in this syndrome is often modest in severity but may be severe and even life-threatening. Major ABO blood group incompatibility is also associated with hemolysis, although this is relatively unusual and generally not severe. Autoimmune hemolytic anemia is a relatively common late complication of allogeneic transplantation and carries significant risk of mortality. Also, alloantibodies may be produced by engrafted cells of the donor's immune system or by residual cells of the patient's immune system following hematopoietic cell transplantation. Hemolysis may occur after solid organ transplantation, particularly as part of the passenger lymphocyte syndrome. PMID- 16041665 TI - Immune hemolytic anemia associated with negative routine serology. AB - Immune hemolytic anemia can occur in patients who have no antibodies detectable by routine procedures (direct [DAT] and indirect [IAT] antiglobulin tests). DAT negative autoimmune hemolytic anemias (AIHAs) represent 5% to 10% of all AIHAs. Three causes have been identified: (1) small numbers of red blood cell (RBC) bound IgG molecules below the threshold of the DAT; (2) IgA and IgM autoantibodies; and (3) low-affinity autoantibodies. Antibody-independent cytotoxic events caused by natural killer (NK) cells have also been implicated. DATs are sometimes found to be positive when tested by reference laboratories, due to poor technique in reading antiglobulin tests in hospital laboratories. Hemolytic transfusion reactions also can occur when no alloantibodies are detectable by routine procedures. In some cases antibodies can be detected by special serologic procedures (such as the Polybrene test); in other instances phenotypically matched RBCs survive well and a specific antigen can be shown to be involved, suggesting a specificity (like anti-C) that is undetectable by any technique. Antibodies other than to blood group antigens, such as human leukocyte antigen (HLA) antibodies, may sometimes be involved. PMID- 16041666 TI - Pathophysiology of hemolytic transfusion reactions. AB - Hemolytic transfusion reactions (HTR) are systemic reactions provoked by immunologic red blood cell (RBC) incompatibility. Clinical and experimental observations of such reactions indicate that they proceed through phases of humoral immune reaction, activation of phagocytes, productions of cytokine mediators, and wide-ranging cellular responses. HTR have many features in common with the systemic inflammatory response syndrome (SIRS). Knowledge of the pathophysiologic mechanisms in HTR suggest that newer biological agents that target complement intermediates or proinflammatory cytokines may be effective agents in the treatment of severe HTRs. PMID- 16041667 TI - Red blood cell alloimmunization in pregnancy. AB - Red blood cell (RBC) alloimmunization in pregnancy continues to occur despite the widespread use of both antenatal and postpartum Rhesus immune globulin (RhIG), due mainly to inadvertent omissions in administration as well as antenatal sensitization prior to RhIG given at 28 weeks' gestation. Additional instances are attributable to the lack of immune globulins to other RBC antigens. Evaluation of the alloimmunized pregnancy begins with the maternal titer. Once a critical value [32 for anti-Rh(D) and other irregular antibodies; 8 for anti-K and -k] is reached, fetal surveillance using serial Doppler ultrasound measurements of the peak velocity in the fetal middle cerebral artery (MCA) is standard. In the case of a heterozygous paternal phenotype, amniocentesis can be performed to detect the antigen-negative fetus that requires no further evaluation. MCA velocities greater than 1.5 multiples of the median necessitate cordocentesis, and if fetal anemia is detected, intrauterine transfusion therapy is initiated. A perinatal survival of greater than 85% with normal neurologic outcome is now expected. Future therapies will target specific immune manipulations in the pregnant patient. PMID- 16041669 TI - Immunoaffinity separation of plasma proteins by IgY microbeads: meeting the needs of proteomic sample preparation and analysis. AB - Separation of complex protein mixtures that have a wide dynamic range of concentration, such as plasma or serum, is a challenge for proteomic analysis. Sample preparation to remove high-abundant proteins is essential for proteomics analysis. Immunoglobulin yolk (IgY) antibodies have unique and advantageous features that enable specific protein removal to aid in the detection of low abundant proteins and biomarker discovery. This report describes the efficiency and effectiveness of IgY microbeads in separating 12 abundant proteins from plasma with an immunoaffinity spin column or LC column. The protein separation and sample preparation process was monitored via SDS-PAGE, 2-DE, LC-MS/MS, or clinical protein assays. The data demonstrate the high specificity of the protein separation, with removal of 95-99.5% of the abundant proteins. IgY microbeads against human proteins can also selectively remove orthologous proteins of other mammals such as mouse, rat, etc. Besides the specificity and reproducibility of the IgY microbeads, the report discusses the factors that may cause potential variations in protein separation such as protein-protein interactions (known as "Interactome"), binding and washing conditions of immunoaffinity reagents, etc. A novel concept of Seppromics is introduced to address methodologies and science of protein separation in a context of proteomics. PMID- 16041670 TI - Do we want our data raw? Including binary mass spectrometry data in public proteomics data repositories. AB - With the human Plasma Proteome Project (PPP) pilot phase completed, the largest and most ambitious proteomics experiment to date has reached its first milestone. The correspondingly impressive amount of data that came from this pilot project emphasized the need for a centralized dissemination mechanism and led to the development of a detailed, PPP specific data gathering infrastructure at the University of Michigan, Ann Arbor as well as the protein identifications database project at the European Bioinformatics Institute as a general proteomics data repository. One issue that crept up while discussing which data to store for the PPP concerns whether the raw, binary data coming from the mass spectrometers should be stored, or rather the more compact and already significantly processed peak lists. As this debate is not restricted to the PPP but relates to the proteomics community in general, we will attempt to detail the relative merits and caveats associated with centralized storage and dissemination of raw data and/or peak lists, building on the extensive experience gained during the PPP pilot phase. Finally, some suggestions are made for both immediate and future storage of MS data in public repositories. PMID- 16041671 TI - PRIDE: the proteomics identifications database. AB - The advent of high-throughput proteomics has enabled the identification of ever increasing numbers of proteins. Correspondingly, the number of publications centered on these protein identifications has increased dramatically. With the first results of the HUPO Plasma Proteome Project being analyzed and many other large-scale proteomics projects about to disseminate their data, this trend is not likely to flatten out any time soon. However, the publication mechanism of these identified proteins has lagged behind in technical terms. Often very long lists of identifications are either published directly with the article, resulting in both a voluminous and rather tedious read, or are included on the publisher's website as supplementary information. In either case, these lists are typically only provided as portable document format documents with a custom-made layout, making it practically impossible for computer programs to interpret them, let alone efficiently query them. Here we propose the proteomics identifications (PRIDE) database (http://www.ebi.ac.uk/pride) as a means to finally turn publicly available data into publicly accessible data. PRIDE offers a web-based query interface, a user-friendly data upload facility, and a documented application programming interface for direct computational access. The complete PRIDE database, source code, data, and support tools are freely available for web access or download and local installation. PMID- 16041672 TI - Plasma Proteome Database as a resource for proteomics research. AB - Plasma is one of the best studied compartments in the human body and serves as an ideal body fluid for the diagnosis of diseases. This report provides a detailed functional annotation of all the plasma proteins identified to date. In all, gene products encoded by 3778 distinct genes were annotated based on proteins previously published in the literature as plasma proteins and the identification of multiple peptides from proteins under HUPO's Plasma Proteome Project. Our analysis revealed that 51% of these genes encoded more than one protein isoform. All single nucleotide polymorphisms involving protein-coding regions were mapped onto the protein sequences. We found a number of examples of isoform-specific subcellular localization as well as tissue expression. This database is an attempt at comprehensive annotation of a complex subproteome and is available on the web at http://www.plasmaproteomedatabase.org. PMID- 16041673 TI - Recent advances in blood-related proteomics. AB - Blood is divided in two compartments, namely, plasma and cells. The latter contain red blood cells, leukocytes, and platelets. From a descriptive medical discipline, hematology has evolved towards a pioneering discipline where molecular biology has permitted the development of prognostic and diagnostic indicators for disease. The recent advance in MS and protein separation now allows similar progress in the analysis of proteins. Proteomics offers great promise for the study of proteins in plasma/serum, indeed a number of proteomics databases for plasma/serum have been established. This is a very complex body fluid containing lipids, carbohydrates, amino acids, vitamins, nucleic acids, hormones, and proteins. About 1500 different proteins have recently been identified, and a number of potential new markers of diseases have been characterized. Here, examples of the enormous promise of plasma/serum proteomic analysis for diagnostic/prognostic markers and information on disease mechanism are given. Within the blood are also a large number of different blood cell types that potentially hold similar information. Proteomics of red blood cells, until now, has not improved our knowledge of these cells, in contrast to the major progresses achieved while studying platelets and leukocytes. In the future, proteomics will change several aspects of hematology. PMID- 16041674 TI - SPS' Digest: the Swiss Proteomics Society selection of proteomics articles. AB - Despite the consolidation of the specialized proteomics literature around a few established journals, such as Proteomics, Molecular and Cellular Proteomics, and the Journal of Proteome Research, a lot of information is still spread in many different publications from different fields, such as analytical sciences, MS, bioinformatics, etc. The purpose of SPS' Digest is to gather a selection of proteomics articles, to categorize them, and to make the list available on a periodic basis through a web page and email alerts. PMID- 16041675 TI - A novel strategy using MASCOT Distiller for analysis of cleavable isotope-coded affinity tag data to quantify protein changes in plasma. AB - A novel strategy consisting of cleavable Isotope-Coded Affinity Tag (cICAT) combined with MASCOT Distiller was evaluated as a tool for the quantification of proteins in "abnormal" patient plasma, prepared by pooling samples from patients with acute stroke. Quantification of all light and heavy cICAT-labelled peptide ion pairs was obtained using MASCOT Distiller combined with a proprietary software. Peptides displaying differences were selected for identification by MS. These preliminary results show the promise of our approach to identify potential biomarkers. PMID- 16041676 TI - Web-based data warehouse on gene expression in human colorectal cancer. AB - Based on biomedical literature databases, we tried a first step for constructing a gene expression "data warehouse" specific to human colorectal cancer (CRC). Results of genome-wide transcriptomic research were available from 12 studies, using various technologies, namely, SAGE, cDNA and oligonucleotide arrays, and adaptor-tagged amplification. Three studies analyzed CRC cell lines and nine studies of human samples. The total number of patients was 144. Out of 982 up- or down-regulated genes, 863 (88%) were found to be differentially expressed in a single study, 88 in two studies, 22 in three studies, 7 in four studies, and only 2 genes in six studies. Eight large-scale proteomics studies were published in CRC, using 2-D-, SDS- or free-flow electrophoresis, involving only 11 patients. Out of 408 differentially expressed proteins, 339 (83%) were found to be differentially expressed only in a single study, 16 in three studies, 10 in four studies, 3 in five, and 1 in eight studies. Confirmation at proteome level of results obtained with large-scale transcriptomics studies was possible in 25%. This proportion was higher (67%) for reproducing proteome results using transcriptomics technologies. Obviously, reproducibility and overlapping between published gene expression results at proteome and transcriptome level are low in human CRC. Thus, the development of standardized processes for collecting samples, storing, retrieving, and querying gene expression data obtained with different technologies is of central importance in translational research. PMID- 16041677 TI - Third-trimester ultrasound dating algorithms derived from pregnancies conceived with artificial reproductive techniques. AB - OBJECTIVE: To derive an accurate formula for ultrasound estimation of gestational age in late pregnancy. METHODS: A database of 123 singleton pregnancies conceived by artificial reproductive techniques with third-trimester ultrasound measurements was studied. Biometry variables included the fetal head circumference (HC) and femur length (FL). The dataset was equally divided into a derivation sample and a target sample. To derive the equations of best fit, regression analysis was used, with true menstrual age as the dependent variable and fetal biometry measurements as independent variables. The formulae were tested on the target set and the menstrual age estimates were compared with the actual menstrual age. Clinical performance was estimated in terms of systematic and random errors, absolute errors and their 95% CI values. RESULTS: The menstrual ages at time of scanning ranged from 26 to 41 weeks, with a mean of 33 weeks. The best performing derived formula was a combination of HC and FL. This had a random error of 7.5 days and prediction errors within a 95% confidence limit of -13 to +17 days. With FL only, the random error was 8.2 days, whereas using the HC only yielded a random error of 9.4 days. CONCLUSIONS: Ultrasound estimation of gestational age in late pregnancy is better than indicated by older publications. It is more accurately estimated by combining the HC with the FL than by using the FL measurements alone. PMID- 16041678 TI - Fetal growth in Peru: comparisons with international fetal size charts and implications for fetal growth assessment. AB - OBJECTIVES: To compare fetal biometry measurements obtained in a Peruvian population with reference fetal size charts obtained in Peruvian and non-Peruvian populations. METHODS: Fetal biometry measurements collected prospectively in 195 uncomplicated pregnancies were included in the presented analysis. At 20, 24, 28, 32, 36 and 38 weeks' gestation, fetal head circumference, abdominal circumference and femur diaphysis length were measured. Fetal biometry measurements were compared with fetal size charts obtained from another Peruvian and two non Peruvian populations from North America and Europe. RESULTS: When compared with ultrasound-based reference fetal size charts obtained from North American and European populations, fetuses from the studied population appeared to grow more slowly with advancing gestational age. This trend was not observed when a Peruvian population, similar to the one studied here, was used as a reference. CONCLUSIONS: The results suggest that fetal growth in this Peruvian population may not be adequately assessed by using reference charts obtained from other populations and have implications for the use of growth standards in antenatal management. PMID- 16041679 TI - Doppler assessment of fetal aortic isthmus blood flow in two different sonographic planes during the second half of gestation. AB - OBJECTIVE: To compare the reliability of Doppler blood flow measurements of the fetal aortic isthmus (AoI) according to whether the sampling plane is obtained from the traditional longitudinal aortic arch (LAA) view or the more recently described three vessels and trachea (3VT) view of the fetal upper mediastinum. METHODS: Doppler blood flow measurements of pulsatility index (PI), resistance index (RI), peak systolic (PSV), end-diastolic (EDV) and time-averaged maximum (TAMXV) velocities were performed in the AoI of 40 fetuses between 24 and 36 weeks of gestation. All measurements were sampled in two different sonographic planes of the AoI: the LAA view, at a few millimeters beyond the origin of the left subclavian artery, and the 3VT view, just before the V-shaped junction of the aortic and ductal arches. All scans were performed by the same observer. The reliability of Doppler blood flow measurements was assessed by calculating intraclass correlation coefficients (ICCs) and limits of agreement between the two different sonographic sites evaluating the AoI. RESULTS: Mean values of PI, RI, PSV, EDV and TAMXV were similar in the LAA and 3VT views. The PI and vascular velocities were reliably measured from both sonographic sites. ICCs for variability of measurements were 0.78, 0.63, 0.63, 0.60 and 0.55 for PI, RI, PSV, EDV and TAMXV, respectively. Limits of agreement revealed minimal disagreement between the two sites of evaluation of the AoI for all measurements. CONCLUSIONS: On the basis of our observations, Doppler blood flow measurements across the fetal AoI can be reliably obtained from both the 3VT and the traditional LAA sonographic views. Since the transverse upper thoracic 3VT plane is achievable in most fetal positions, Doppler study of the AoI appears to be easier than expected. PMID- 16041680 TI - Monoamniotic twins discordant for anencephaly managed conservatively with good outcomes: two case reports and a review of the literature. AB - Monoamniotic twin pregnancy discordant for anencephaly (MATDA) is a rare occurrence with only seven prior reported cases. Selective termination has been advocated in managing discordant monoamniotic twins. We report two cases managed expectantly with good outcomes and review other previously reported cases. The first case was a primigravid woman diagnosed with MATDA at 18 weeks. She was managed expectantly until 32 + 5 weeks when a Cesarean section was performed for preterm labor. The surviving female infant weighed 1610 g. The second case was a multigravid woman who was diagnosed with MATDA at 17 + 5 weeks and was managed as an outpatient. An emergency Cesarean section was performed at 31 weeks for non reassuring monitoring and the surviving male infant weighed 1790 g. In both cases, the survivors were discharged home in good condition. A review of these two cases and those in the literature suggests that expectant management should be considered among management options for this rare condition. PMID- 16041681 TI - Papillary adenofibroma of the cervix: a case report. AB - Adenofibroma is an extremely rare benign biphasic neoplasm that is classified into the mixed epithelial and mesenchymal tumor group. It typically affects the endometrium, but may occur in the cervix or in an extrauterine location. Preoperative diagnosis of this tumor is usually difficult. We describe the case of a 55-year-old woman with papillary cervical adenofibroma, which appeared as a cervical mass containing multiple cystic components on transvaginal ultrasound. This lesion appears to be clinically and histologically benign but must be differentiated from malignant lesions of the uterus, particularly from adenosarcoma, which can be suggestive of adenofibroma. Accurate diagnosis of these benign tumors permits appropriate counseling of patients. PMID- 16041682 TI - DISQ 2: Endometrial thickness in asymptomatic women. PMID- 16041683 TI - Prenatal diagnosis of fetal genital prolapse. PMID- 16041684 TI - Fetal growth: towards an international standard. PMID- 16041685 TI - Sequential segmental analysis in complex fetal cardiac abnormalities: a logical approach to diagnosis. PMID- 16041686 TI - Dilution of reporter gene with stuffer DNA does not alter the transfection efficiency of polyethylenimines. AB - BACKGROUND: Polyethylenimines (PEIs) are among the most efficient non-viral gene transfer agents developed so far. However, transfections with these polymers were shown to require a very high copy number of plasmid DNA per cell to achieve gene expression. Here, we investigate whether it is possible to reduce the amount of plasmid DNA while keeping a high transfection efficiency. METHODS: Transfection experiments were performed under various conditions in order to study the interdependence between the amount of reporter DNA, the amine-to-phosphate ratio and the transfection efficiency. RESULTS: When suboptimal amounts of linear PEI 22 kDa/DNA complexes were used for transfection, a severe reduction in reporter gene expression was observed. On the other hand, for optimal amounts of PEI/DNA complexes more than half of the reporter gene can be replaced by carrier DNA or polyglutamic acid without substantially decreasing the transfection efficiency of the polymer both in cultured cells and after systemic administration in mice. When used under the same in vitro experimental conditions, the lipospermine DOGS, but not the monocationic lipid DOTAP, gave similar results. CONCLUSIONS: Taken together, our data suggest that the activity of compounds with endosome-buffering capacities, such as PEIs and lipospermines, requires a threshold amount of transfection agent. In addition, our results indicate that, in many gene transfer situations, it will be possible to lower the dose of active plasmid thus reducing costs and the risk of immune stimulation triggered by bacterial DNA. PMID- 16041688 TI - Ultrasound screening for gestational trophoblastic disease. PMID- 16041689 TI - Sustained elevation of neutrophils in rats induced by lentivirus-mediated G-CSF delivery. AB - BACKGROUND: Patients with severe chronic and cyclic neutropenia, characterized by neutrophil numbers <500 cells/microl, are treated daily with recombinant granulocyte colony-stimulating factor (G-CSF). As an alternative delivery approach we investigated the ability of lentivirus vectors to provide sustained G CSF expression. METHODS: Fischer rats were injected intramuscularly (IM) with vesicular stomatitis virus G (VSV-G)-pseudotyped lentivirus pRRL-CMV-G-CSF-SIN that encoded rat G-CSF cDNA regulated by the human cytomegalovirus (CMV) promoter and incorporated a self-inactivating (SIN) construct in the 3' long terminal repeat (LTR). Control rats received normal saline or lentivirus encoding the enhanced green fluorescent protein (eGFP). Rats were serially monitored for blood cell production and tissues assayed for provirus distribution. RESULTS: Rats receiving a single IM injection of lentivirus exhibited elevated neutrophil counts for 14 months. Virus administration of 6 x 10(7) infectious units induced sustained levels of neutrophil production having a mean +/- standard deviation (SD) of 5650 +/- 900 cells/microl and rats that received a 10-fold lower dose of virus showed mean neutrophil counts of 3340 +/- 740 cells/microl. These were significantly higher than the mean of control animals receiving saline or control lentivirus (1,760 +/- 540 cells/microl, P < 0.0001). White blood cell (WBC) counts were significantly elevated in treated over control animals (P < 0.0001). Hematocrits (P > 0.3), lymphocytes (P > 0.2) and platelets (P > 0.1) were not significantly different between control and treated animals. Genomic DNA from muscle at the injection sites was positive for provirus, whereas lung, spleen, liver, kidney and non-injected muscle samples were all negative, suggesting lack of virus spread. CONCLUSIONS: These studies indicate that lentivirus vectors administered IM provide sustained, therapeutic levels of neutrophils and suggest this approach to treat patients with severe and cyclic neutropenia. PMID- 16041690 TI - Evidence that DL-3-aminobutyric acid and acibenzolar-S-methyl induce resistance against bacterial head rot disease of broccoli. AB - Head rot of broccoli caused by Pseudomonas marginalis (Brown) Stevens and P. fluorescens Migula is a major disease in Brittany (France). To date, no accession with a satisfactory field resistance has been identified, and available pesticides are not effective in controlling the disease. The aim of this study was to test whether acibenzolar-S-methyl (ASM), DL-3-aminobutyric acid (BABA) and potassium phosphonate (K(2)HPO(3)), known to induce resistance against various diseases, can help protect broccoli against head rot. The susceptible broccoli F1 hybrids Marathon and Shogun were grown in a greenhouse until head formation. They were then sprayed with ASM (0.23 mM AI), BABA (20 mM AI) or potassium phosphonate (37.41 mM AI) until runoff. In one experiment, heads from treated plants were excised, inoculated (10(4) cfu ml(-1)) and incubated in Magenta GA7 vessels. In another experiment, heads were inoculated on treated living plants. Disease ratings were made 5 days after inoculation. Antibiotic- and water-treated plants served as controls. Results obtained showed that, on excised treated heads, potassium phosphonate was not protective and disease scores were comparable with those of the water control. BABA- and ASM-treated excised heads were poorly, but significantly, protected. On whole plants with heads attached, the latter two compounds were much more effective. ASM-induced resistance increased in effectiveness over 8 days after inoculation, whilst that induced by BABA decreased. This result suggests that testing disease resistance inducers on excised broccoli heads is not accurate. ASM and BABA may offer alternative methods for controlling head rot of broccoli. PMID- 16041691 TI - Up-regulated expression of ADAM17 in human colon carcinoma: co-expression with EGFR in neoplastic and endothelial cells. AB - The ADAM17 metalloproteinase (a disintegrin and metalloprotease 17) controls epidermal growth factor receptor (EGFR) activation through regulated shedding of EGFR ligands. With the advent of new therapeutic options targeting EGFR signalling in colon carcinoma, it was decided to determine ADAM17 status in relation to clinico-pathological parameters and EGFR status. To this end, a series of 39 colon carcinomas were analysed. Immunohistochemistry and immunofluorescence were used to localize ADAM17, EGFR, and the activated forms of EGFR. The activated form of ADAM17 was assessed in primary cancers and colon cell lines by immunoblotting. ADAM17 and EGFR mRNA levels were assessed by quantitative RT-PCR. Chromogenic in situ hybridization (CISH) was used to quantify the HER1 gene. ADAM17 was strongly expressed in all tumours, by both neoplastic and endothelial cells. It was expressed both as a pro- and as an active form in tumours and colonic cancer cell lines. ADAM17 mRNA was up regulated in 90% of colon carcinomas relative to the paired normal mucosa, whatever the tumour grade or stage. When present, activated EGFR was co-expressed with ADAM17 by colon carcinomas, although at a variable level among tumour cells, and by endothelial cells. EGFR mRNA was overexpressed in 77% of colon carcinomas compared with the paired normal mucosa. One case showed high-level amplification of HER1. In conclusion, this study is the first demonstration that ADAM17 is overexpressed in human primary colon carcinoma, whatever the tumour stage and differentiation and whatever the level of EGFR expression. Its co-expression with EGFR, in both neoplastic and endothelial cells, suggests a role for ADAM17 in tumour growth and angiogenesis. PMID- 16041692 TI - Airway proteoglycans are differentially altered in fatal asthma. AB - It has been suggested that airway remodelling is responsible for the persistent airway obstruction and decline in lung function observed in some asthmatic patients. The small airways are thought to contribute significantly to this functional impairment. Proteoglycans (PGs) are important components of the extracellular matrix (ECM) in the lungs. Besides controlling biophysical properties of the ECM, they play important roles in the regulation of some cytokines. Increased subepithelial PG deposition in the airways of mild asthmatics has been reported. However, there are no data on the PG content in small airways in asthma. This study has compared the content and distribution of PGs in large and small airways of patients who died of asthma with those in control lungs. Immunohistochemistry and image analysis were used to determine the content of lumican, decorin, biglycan, and versican in large (internal perimeter >6 mm) and small (internal perimeter < or =6 mm) airways of 18 patients who had died of asthma (A) and ten controls (C). The results were expressed as PG area (microm2)/epithelial basement membrane length (microm). The main differences between asthmatics and controls were observed in the small airways. There was a significant decrease in decorin and lumican contents in the external area of small airways in asthmatics (decorin: A = 1.05 +/- 0.27 microm, C = 3.97 +/- 1.17 microm, p = 0.042; lumican: A = 1.97 +/- 0.37 microm, C = 5.66 +/- 0.99 microm, p = 0.002). A significant increase in versican content in the internal area of small and large airways in asthmatics was also observed (small: A = 7.48 +/- 0.84 microm, C = 5.16 +/- 0.61 microm, p = 0.045; large: A = 18.38 +/- 1.94 microm, C = 11.90 +/- 2.86 microm, p = 0.028). The results show that PGs are differentially expressed in the airways of fatal asthma and may contribute to airway remodelling. These data reinforce the importance of the small airways in airway remodelling in asthma. PMID- 16041693 TI - Chromosomal imbalances in wood dust-related adenocarcinomas of the inner nose and their associations with pathological parameters. AB - Comparative genomic hybridization (CGH) was used to screen 42 wood dust-related sinonasal adenocarcinomas for chromosomal alterations. The tumour collection comprised 39 papillary-tubular cylinder cell adenocarcinomas (PTCCs; six cases G1, 23 G2, and ten G3), two alveolar goblet cell adenocarcinomas (AGCs), and one signet ring cell adenocarcinoma (SRC), according to the Kleinsasser and Schroeder classification. Copy number changes were detected in 41 tumours (97.6%). The one carcinoma without imbalances was a PTCC-G1. DNA gains were most frequently seen on chromosomes 12p (83%), 7q (74%), 8q (71%), and 20q (71%), 11q (61%), 22 (59%), and 1q (52%). Pronounced overrepresentations suggestive of high copy amplifications were detected on 8q (15 cases, 36%), 7q (six cases, 14%), 20q (five cases, 12%), 13q14 (three cases, 7%), 1q22, 5p, 12p and 20 (two cases, 5% each), and 2q24, 3q13, 3q22, 7p, 14q12, and 16q13 (one case, each 2%). Frequent chromosomal losses occurred at 5q (81%), 18q (76%), 4 (74%), 8p (61%), 9p (60%), 6q and 17p (52% each), and 3p, 13q, and 21 (50% each). There was a quantitative as well as a qualitative increase of alterations from PTCC-G1 to PTCC-G2 and finally PTCC-G3, confirming the usefulness of histopathological grading. While PTCC-G1 carried only a few alterations, namely gains on chromosomes 17 and 7 as well as losses of 4q and 13q, PTCC-G2 already carried many of the above-mentioned alterations, while PTCC-G3 showed significantly more gains of 7q, 8q, and 12p, and losses of 8p and 17p. Additionally, the latter subgroup was particularly prone to carry pronounced DNA gains. These data provide further evidence for a recurrent pattern of chromosomal imbalances in sinonasal adenocarcinomas and highlight distinct aberrations that are associated with tumour differentiation and progression. PMID- 16041694 TI - Protective role of heat shock protein 27 in gastric mucosal injury. AB - Heat shock protein 27 (HSP27) mediates cytoprotective effects through its function as a molecular chaperone and through the phosphorylation-dependent stabilization of actin filaments. The role of HSP27 in gastric ulcer formation and healing is, however, unknown. The expression of HSP27 was studied in human gastric tissue specimens obtained from patients with gastric ulcers and from healthy Helicobacter pylori-negative individuals, who received low-dose aspirin, rofecoxib, and the combination in a prospective study. The susceptibility of the gastric mucosa to indomethacin-induced lesions was further studied in transgenic mice overexpressing human HSP27 (Tg(huHSP27)) and compared with wild-type mice (Wt). The expression of HSP27, COX-1, and COX-2 was investigated in Tg(huHSP27) mice and Wt mice by immunohistochemistry and western blot analysis. While no specific changes in HSP27 expression were found following exposure of healthy human gastric mucosa to oral administration of aspirin or refocoxib, chronic gastric ulcers showed strong HSP27 expression at the ulcer base and margins. Here it was expressed by granulation tissue and regenerating surface epithelium. In Tg(huHSP27) mice, overexpression of HSP27 led to a significant decrease of indomethacin-induced erosions and ulcers compared with Wt mice. COX-1 and COX-2 levels did not change. HSP27 is involved in chronic gastric ulcer repair mechanisms in humans, while overexpression of human HSP27 in gastric epithelial cells in mice reduces the susceptibility to non-steroidal anti-inflammatory drug (NSAID)-induced gastric ulceration. This indicates that HSP27 expression is critical for mucosal protection in the stomach. PMID- 16041695 TI - BIC and miR-155 are highly expressed in Hodgkin, primary mediastinal and diffuse large B cell lymphomas. AB - In a previous study we demonstrated high expression of the non-coding BIC gene in the vast majority of Hodgkin's lymphomas (HLs). Evidence suggesting that BIC is a primary microRNA transcript containing the mature microRNA-155 (miR-155) as part of a RNA hairpin is now accumulating. We therefore analysed HL cell lines and tissue samples to determine whether miR-155 is also expressed in HL. High levels of miR-155 could be demonstrated, indicating that BIC is processed into a microRNA in HL. Most non-HL subtypes were negative for BIC as determined by RNA ISH. However, in diffuse large B cell lymphoma (DLBCL) and primary mediastinal B cell lymphoma (PMBL), significant percentages of positive tumour cells were observed in 12/18 and 8/8 cases. A higher proportion of tumour cells were positive for BIC in DLBCL with activated B cell-like phenotype than in DLBCL with germinal centre B cell-like phenotype. Differential BIC expression was confirmed by qRT-PCR analysis. Northern blot analysis showed expression of miR-155 in all DLBCL and PMBL derived cell lines and tissue samples analysed. In summary, we demonstrate expression of primary microRNA BIC and its derivative miR-155 in HL, PMBL and DLBCL. PMID- 16041696 TI - Mortality and F1 progeny of the lesser grain borer, Rhyzopertha dominica (F), on wheat treated with diatomaceous earth: effects of rate, exposure period and relative humidity. AB - A series of experiments at 25 (+/-1) degrees C were conducted in which different application rates of diatomaceous earth (DE) formulation Protect-It at two levels of relative humidity, 40 and 55%, and at three exposure periods were evaluated for control of Rhyzopertha dominica (F). Test insects were placed in vials containing 40 g of soft winter wheat mixed with 0.25, 0.50, 1.00, 1.50 and 2.00 g DE kg(-1). At all rates except 0.25 g kg(-1), mortality was significantly different from that in the control at the relevant exposure period. After each exposure interval, dead and live insects were counted and removed, and the vial containing wheat was then returned to the appropriate humidity chamber for 8 weeks until F(1) adults emerged. F(1) progeny production was significantly different from the control group at all rates. Mortalities for 1, 2 and 3 weeks exposure were found to be 47 (+/-5)%. Despite the fact that mortality increased with increasing rate, total mortality was not achieved even at the highest rate of DE. F(1) progeny production decreased with increasing rate for both RH conditions, and containment of population for both RH conditions was achieved at 1.00 g DE kg(-1). For each exposure period, F(1) progeny production of R dominica decreased with increasing rate of DE, and population suppression was achieved at 1.00 g DE kg(-1) for all exposure intervals. In summary, Protect-It resulted in reduced F(1) adult progeny and containment of population was achieved at 1.00 g DE kg(-1) at which rate mortality was 77%. PMID- 16041697 TI - Quantitative study on selective stacking of zwitterions in large-volume sample matrix by moving reaction boundary in capillary electrophoresis. AB - The paper advanced the theoretical procedures for quantitative design on selective stacking of zwitterions in full capillary sample matrix by a cathodic direction moving reaction boundary (MRB) in capillary electrophoresis (CE) under control of electroosmotic flow (EOF). With the procedures, we conducted the theoretical computations on the selective stacking of two test analytes of L histidine (His) and L-tryptophan (Trp) by the MRB created with 30 mM pH 3.0 formic acid-NaOH buffer and 2-80 mM sodium formate. The results revealed the following three predictions. At first, the MRB cannot stack His and Trp plugs if less than 12.5 mM sodium formate is used to form the MRB and prepare the sample matrix. Second, the MRB can stack His and/or Trp sample plugs completely if higher than 50 mM sodium formate is chosen to form the MRB. Third, the MRB can only focus His plug completely, but stack Trp plug partially if 20-50 mM sodium formate is used; this implied the complete MRB-induced selective stacking to His rather than Trp. All the three predictions were quantitatively proved by the experiments. With great dilution of sample matrix and control of EOF, controllable, simultaneous and MRB-induced selective stacking and separation of zwitterions were achieved. The theoretical results hold evident significances to the quantitative design of selective stacking conditions and the increase of detection sensitivity of zwitterions in CE. In addition, the control of EOF by cetyltrimethylammonium bromide (CTAB) can evidently improve the stacking efficiency to both His and Trp. PMID- 16041698 TI - "In-gel patch electrophoresis:" a new method for environmental DNA purification. AB - Most of the microorganism species are largely untapped and could represent an interesting reservoir of genes useful for biotechnological applications. Unfortunately, a major difficulty associated with the methods used to isolate environmental DNA is related to the contamination of the extracted material with humic substances. These polyphenolic compounds inhibit the DNA processing reactions and severely impede cloning procedures. In this work, we describe a rapid, simple, and efficient method for the purification of genomic DNA from environmental samples: we added a chromatography step directly embedded into an agarose gel electrophoresis. This strategy enabled the DNA extraction from various environmental samples and it appeared that the purity grade was compatible with digestion by restriction enzymes and polymerase chain reaction (PCR) amplifications. PMID- 16041699 TI - Detection of single-base mutation by affinity capillary electrophoresis using a DNA-polyacrylamide conjugate. AB - We have developed an affinity capillary electrophoresis (ACE) method for detection of gene point mutations using a DNA-polyacrylamide conjugate as a pseudostationary affinity phase. In this study, the target DNA was prepared by mixing two PCR products: the wild type of K-ras gene and its codon 12 point mutant. The ligand DNA was designed to be complementary to codons 11 and 12 of the wild type. The target DNA was denatured by the addition of formamide and by heating at 95 degrees C for 5 min, and then electrophoretically separated by difference in affinity to the pseudoimmobilized ligand DNA. The method successfully separated a mixture of the wild-type DNA and each of six codon 12 point mutants by the same ligand DNA. The limit of mutation detection was determined by mixing the wild-type DNA with decreasing concentrations of the mutant DNA. The lowest level of detection was 10% mutant DNA in a background of the wild type. The practicability of this method has been confirmed using a colorectal carcinoma cell line. This study is the first demonstration of detection of gene point mutation in polymerase chain reaction (PCR) products using ACE, and opens up a new possibility of CE-based gene diagnosis. PMID- 16041700 TI - Modified Hadamard transform microchip electrophoresis. AB - Sensitivity is a crucial point in the development applications for medicine or environmental samples in which the analytes are present in the nanomolar range. Besides further technical development of detection systems, the multiplex sample injection technique can be applied for enhancing the signal-to-noise ratio. Hadamard transform is easily applied to microchip electrophoresis due to the fact that sample injection is generally achieved through cross, double-tee, or tee injector structures. This paper reports the first demonstration of a modified Hadamard transform electrophoresis on a microchip by using an amperometric detector. Contrary to the previous Hadamard applications, the resolution (number of points per unit of time) of electropherograms obtained is independent of the number of injections. PMID- 16041701 TI - Nanoparticle-filled capillary electrophoresis for the separation of long DNA molecules in the presence of hydrodynamic and electrokinetic forces. AB - We report the analysis of long DNA molecules by nanoparticle-filled capillary electrophoresis (NFCE) under the influences of hydrodynamic and electrokinetic forces. The gold nanoparticle (GNP)/polymer composites (GNPPs) prepared from GNPs and poly(ethylene oxide) were filled in a capillary to act as separation matrices for DNA separation. The separations of lambda-DNA (0.12-23.1 kbp) and high molecular-weight DNA markers (8.27-48.5 kbp) by NFCE, under an electric field of 140 V/cm and a hydrodynamic flow velocity of 554 microm/s, were accomplished within 5 min. To further investigate the separation mechanism, the migration of lambda-DNA was monitored in real time using a charge-coupled device (CCD) imaging system. The GNPPs provide greater retardation than do conventional polymer media when they are encountered during the electrophoretic process. The presence of interactions between the GNPPs and the DNA molecules is further supported by the fluorescence quenching of prelabeled lambda-DNA, which occurs through an energy transfer mechanism. Based on the results presented in this study, we suggest that the electric field, hydrodynamic flow, and GNPP concentration are the three main determinants of DNA separation in NFCE. PMID- 16041702 TI - Analyses of phenolic compounds by microemulsion electrokinetic chromatography. AB - In this study, a microemulsion electrokinetic chromatography (MEEKC) method was developed to analyze and detect 13 phenolic compounds (syringic acid, p-cumaric acid, vanillic acid, caffeic acid, gallic acid, 3,4-dihydroxybenzoic acid, 4 hydroxybenzoic acid, (+)-catechin, (-)-epigallocatechin, (-)-epicatechin gallate, (-)-epigallocatechin gallate, (-)-epicatechin, and (-)-gallocatechin), which are present in many plant-derived foods. The effects of cosurfactant, organic modifier, and oil were examined in order to optimize the separation of these phenolic compounds. The amounts of cosurfactant (cyclohexanol) and organic modifier (acetonitrile) were determined as the major influence on the separation selectivity, while the type of oil partially affected the separation resolution of the phenolic compounds. A highly efficient MEEKC separation method was achieved within 14 min by using a microemulsion solution of pH 2.0 containing 2.89% w/v SDS, 1.36% w/v heptane, 7.66% w/v cyclohexanol, and 2% w/v ACN. Furthermore, the present work could demonstrate that the nature of the oil phase has a significant influence on the separation selectivity of phenolic compounds. PMID- 16041703 TI - Poly(methylmethacrylate) and Topas capillary electrophoresis microchip performance with electrochemical detection. AB - A capillary electrophoresis (CE) microchip made of a new and promising polymeric material: Topas (thermoplastic olefin polymer of amorphous structure), a cyclic olefin copolymer with high chemical resistance, has been tested for the first time with analytical purposes, employing an electrochemical detection. A simple end-channel platinum amperometric detector has been designed, checked, and optimized in a poly-(methylmethacrylate) (PMMA) CE microchip. The end-channel design is based on a platinum wire manually aligned at the exit of the separation channel. This is a simple and durable detection in which the working electrode is not pretreated. H(2)O(2) was employed as model analyte to study the performance of the PMMA microchip and the detector. Factors influencing migration and detection processes were examined and optimized. Separation of H(2)O(2) and L ascorbic acid (AsA) was developed in order to evaluate the efficiency of microchips using different buffer systems. This detection has been checked for the first time with a microchip made of Topas, obtaining a good linear relationship for mixtures of H(2)O(2) and AsA in different buffers. PMID- 16041704 TI - A family of single-isomer positively charged cyclodextrins as chiral selectors for capillary electrophoresis: mono-6A-butylammonium-6A-deoxy-beta-cyclodextrin tosylate. AB - A novel single-isomer positively charged beta-cyclodextrin (beta-CD), mono-6(A) butylammonium-6(A)-deoxy-beta-cyclodextrin tosylate (BuAM-beta-CD), has been synthesized, characterized, and used for the enantioseparations of alpha-hydroxy acids, carboxylic acids, and ampholytic analytes by capillary electrophoresis in acidic aqueous background electrolytes. The effective mobilities of all studied analytes decreased with increasing concentration of CD. Satisfactory resolutions were obtained for alpha-hydroxy acids over a wide range of chiral selector concentration. The optimum CD concentration was lower than 5 mM for the carboxylic acids, while higher than 20 mM for alpha-hydroxy acids. Inclusion complexation in combination with ion pair interaction seemed to account for the chiral discrimination process. The hydrogen bonding may provide secondary contribution for the chiral resolution of alpha-hydroxy acids. In addition, BuAM beta-CD was further proved to be an effective chiral selector for anionic analytes by the baseline enantioseparation of a six-acid mixture within 20 min. PMID- 16041705 TI - Isoelectric focusing in serial immobilized pH gradient gels to improve protein separation in proteomic analysis. AB - We previously demonstrated the separation of proteins by isoelectric focusing (IEF) over pH 4-8 immobilized pH gradients (IPGs) over 54 cm (Poland et al., Electrophoresis 2003, 24, 1271). Here we show that similar results can be conveniently achieved using commercially available IPGs of appropriate pH ranges positioned end-on-end in series during electrophoresis, which we term "daisy chain IEF". Proteins efficiently electrophorese from one IPG to another during IEF by traversing buffer-filled porous bridges between the serial IPGs. A variety of materials can function as bridges, including paper, polyacrylamide gels or even IPGs. The quality of two-dimensional (2-D) protein patterns is not apparently worse than that generated by conventional IEF using the same individual IPGs. A major advantage of this method is that sample is consumed efficiently, without the requirement for preliminary steps, such as chamber IEF. This advantage is pronounced when working with extremely limited sources of samples, such as with clinical biopsies or cellular subfractions. The present study was limited by the commercial availability of suitable pH gradients. Proteomics analyses could be further improved if commercial vendors would manufacture IPGs with suitable pH ranges to achieve high resolution (approximately 100 cm) IEF separation of proteins in one electrophoretic step over the pH range 2-12. PMID- 16041706 TI - Separation of DNA fragments for fast diagnosis by microchip electrophoresis using programmed field strength gradient. AB - We evaluated a novel strategy for fast diagnosis by microchip electrophoresis (ME), using programmed field strength gradients (PFSG) in a conventional glass double-T microfluidic chip. The ME-PFSG allows for the ultrafast separation and enhanced resolving power for target DNA fragments. These results are based on electric field strength gradients (FSG) that use an ME separation step in a sieving gel matrix poly-(ethylene oxide). The gradient can develop staircase or programmed shapes FSG over the time. The PFSG method could be easily used to increase separation efficiency and resolution in ME separation of specific size DNA fragments. Compared to ME that uses a conventional and constantly applied electric field (isoelectrostatic) method, the ME-PFSG achieved about 15-fold faster analysis time during the separation of 100 bp DNA ladder. The ME-PFSG was also applied to the fast analysis of the PCR products, 591 and 1191 bp DNA fragments from the 18S rRNA of Babesia gibsoni and Babesia caballi. PMID- 16041707 TI - Peak shape modeling by Haarhoff-Van der Linde function for the determination of correct migration times: a new insight into affinity capillary electrophoresis. AB - Among the different experimental strategies available in capillary electrophoresis (CE) to determine binding parameters, affinity capillary electrophoresis (ACE) has been the most widely embraced due to its easiness of implementation and of data handling. Ligand-substrate binding constants are thus directly derived from the substrate migration time shifts resulting from the variation of ligand concentration introduced in a background electrolyte. Classically, the substrate migration time is measured on top of the electrophoretic peak, assuming symmetrical peak shape. Depending on both substrate and ligand concentrations that may be required to meet detection sensitivity or complexation conditions, zonal migrations in ACE may, however, produce triangular peak shape, most often due to pronounced electromigration dispersion (EMD), and this may result in positively or negatively erroneous migration time assessments. In this work, EMD distorted triangular peak shapes obtained in the course of host-guest complexation studies were fitted with the Haarhoff-Van der Linde function, allowing better estimation of migration time. The model systems studied were those of beta-cyclodextrin and naproxen, 2 naphthalenesulfonate, or 1-adamantanecarboxylate. The impact of this correction on binding isotherms and binding constant evaluation was exemplified. Furthermore, in situations where the substrate concentration injected by far overtakes that of the ligand in the electrolyte, the interest in this peak shape correction was discussed in connection with the question of whether the free ligand concentration can be still considered equal to the ligand concentration introduced, a question that still remains under debate nowadays. PMID- 16041708 TI - Dye-free protein molecular weight markers. AB - Protein molecular weight markers are widely used in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Here, we describe novel protein molecular weight markers in which a prestaining procedure is no longer needed. Green fluorescent protein (GFP) is stable and resistant to denaturing agents/conditions. Various histidine-tagged GFP fusion proteins were overexpressed in Escherichia coli and purified by metal affinity chromatography. The minimal amount of each protein marker needed for analysis in SDS-PAGE and Western blot under visible light was 62.5 and 125 ng, respectively. Under ultraviolet (UV) ray, the minimal amount of each protein marker needed for analysis in SDS-PAGE and Western blot was half of those amounts used under visible light, respectively. Collectively, the accuracy, sensitivity, ease, economy, and flexibility of our strategy may reinforce the application of GFP in molecular biology. PMID- 16041709 TI - Poly(methacrylic acid-ethylene glycol dimethacrylate) monolith in-tube solid phase microextraction applied to simultaneous analysis of some amphetamine derivatives in urine by capillary zone electrophoresis. AB - A method based on in-tube solid-phase microextraction and capillary zone electrophoresis (CZE) was proposed for simultaneously determining four amphetamines (amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine, and 3,4-methylenedioxymethamphetamine) in urine. A poly(methacrylic acid-ethylene glycol dimethacrylate) monolithic capillary column, which can provide sufficient extraction efficiency, was introduced for the extraction of amphetamines from urine samples. The hydrophobic main chains and acidic pendant groups of the monolithic column make it a superior material for extraction of basic analytes from aqueous matrix. After extraction, the samples were analyzed by CZE. The best separation was achieved using a buffer composed of 0.1 M disodium hydrogen phosphate (adjusted to pH 4.5 with 1 M hydrochloric acid) and 20% methanol v/v, with a temperature and voltage of 25 degrees C and 20 kV, respectively. By applying electrokinetic injection with field-amplified sample stacking, detection limits of 25-34 microg/L were achieved. Excellent method of reproducibility was found over a linear range of 0.1-5 mg/L. Determination of these analytes from abusers' urine sample was also demonstrated. PMID- 16041710 TI - Retention behavior of proteins in size-exclusion electrochromatography with a low voltage electric field perpendicular to the liquid phase streamline. AB - A novel preparative size-exclusion electrochromatography with an oscillatory low voltage electric field perpendicular to the liquid phase streamline (pSEEC) was proposed with a column design of rectangular cross-section. The column of 12 cm length was packed with Sephadex G-75, and the retention behavior of bovine serum albumin (BSA) and myoglobin (Myo) was extensively investigated under various conditions. The results indicated that the partition coefficient of a charged protein increased significantly on increasing the current strength as well as the difference between its pI and pH. The partition coefficient also increased on decreasing the mobile phase conductivity. For the gel-excluded protein like BSA, the concentration polarization (CP) on the gel surface induced by the protein electromigration was the main reason for the increased retention. For a gel permeable protein like Myo, both the CP and electrophoretic migration in the solid phase contributed to its increased retention. Further results exhibited that the polarization would be offset by diffusion, because the accumulated protein would flux back to the bulk liquid phase. Therefore, when the electrically induced mass flux was equal to the diffusion flux, the partition coefficient did not change with a further increase of the oscillatory current cycle. Finally, pSEEC was compared with SEC in the separation of protein mixtures of BSA/Myo as well as BSA/Myo/lysozyme. The results showed much better resolutions of the protein mixtures in pSEEC with the column as short as 12 cm. The potential of pSEEC for preparative protein separation was therefore demonstrated. PMID- 16041711 TI - Efficacy and fate of glyphosate on Swedish railway embankments. AB - The herbicide glyphosate, N-(phosphonomethyl)glycine, as Spectra (240 g AI litre( 1) SL; Monsanto Europe AB), RoundUp (360 g AI litre(-1) SL; Monsanto) and RoundUp Bio (360 g AI litre(-1) SL; Monsanto), have been used for weed control on Swedish railway embankments since 1986. This article summarizes results from studies of the weed effect and behaviour of glyphosate for the period 1984-2003. Studies on a railway embankment with a range of application rates showed excellent weed control at 5 litre ha(-1) of RoundUp Bio. The appearance of glyphosate and its metabolite AMPA [(aminomethyl)phosphonic acid] in the embankment, eg mobility and persistence, was also studied. Mobility was low in most cases, the main proportion of both glyphosate and AMPA being found in the upper 30-cm layer although minor amounts penetrated to lower depths. The 50% disappearance time of glyphosate was generally <5 months in railway embankments but cases with longer persistence were found. Transport to the groundwater was observed for glyphosate and AMPA in groundwater pipes along tracks. Downward transport appears to be dependent on the application rate, which should not exceed 3 litre ha(-1) of RoundUp Bio to avoid groundwater contamination. A lower rate of glyphosate mixed with a low rate of another herbicide may achieve acceptable weed control and be environmentally safer. PMID- 16041712 TI - Effect of freezing and thawing on microbial activity and glyphosate degradation in two Norwegian soils. AB - Little research has been done on pesticide dissipation in cold climates and there is a need to focus on the influence of climate on pesticide degradation in soil. Glyphosate, N-(phosphonomethyl)glycine, is a herbicide frequently used for controlling perennial weeds through application after harvest and was used as a model compound for this study. The effect of freeze-thaw activity on the availability of glyphosate in soil, and consequently its mineralization by soil microorganisms, was studied through laboratory incubations of repacked soil cores treated with 14C-labelled glyphosate and subjected to different freeze-thaw treatments. Winter simulation regimes applied were constant thaw (+5 degrees C), constant freezing (-5 degrees C), unstable conditions with short fluctuations (24 h of -5 degrees C followed by 24 h of +5 degrees C), and long duration fluctuations (3 weeks of -5 degrees C followed by 3 weeks of +5 degrees C). Distribution of 14C-glyphosate was followed during the incubation through measurements of the mineralized fraction (14CO2), soil water fraction, KOH extractable fraction, and non-extractable fraction. Microbial parameters used to characterize the soils were estimates of size of microbial biomass, overall microbial activity and microbial diversity. The constant freezing treatment exhibited the lowest amount of glyphosate mineralization. The constant thawed treatment and the treatments with fluctuating temperature exhibited significantly increased mineralization. These results were in accordance with the observed concentration of glyphosate in soil water; the higher the activity, the lower the concentration. The amount of glyphosate extractable with KOH and the resulting non-extractable fraction, however, were not significantly affected by soil type or temperature regime. The glyphosate mineralization pattern was comparable with the overall microbial activity in the soils. Observed different levels of diversity might explain some of the difference in total glyphosate mineralization between soils. PMID- 16041713 TI - Functional expression of the peptide transporter PEPT2 in the mammalian enteric nervous system. AB - The peptide transporter PEPT2 mediates transmembrane uptake of small peptides. So far, its expression has not been evidenced in the gastrointestinal tract. We have investigated peptide transport activity in the neuromuscular layers of the gastrointestinal tract by using the fluorescent tracer-dipeptide beta-Ala-Lys Nepsilon-7-amino-4-methyl-coumarin-3-acetic acid (Ala-Lys-AMCA). Whole-mount preparations from mouse, rat, and guinea pig stomach and small and large intestine were incubated with Ala-Lys-AMCA in the presence or absence of the uptake-inhibitors L-histidine, D-phenylalanyl-L-alanine (D-Phe-Ala), glycyl-L sarcosine (Gly-Sar), glycyl-L-glutamine (Gly-Gln), benzylpenicillin, and cefadroxil. Fluorescence microscopy revealed that Ala-Lys-AMCA specifically accumulated in both ganglionic layers of the enteric nervous system (ENS) in all regions and species studied. This could be inhibited by Gly-Sar, D-Phe-Ala, Gly Gln, and cefadroxil, but not by free histidine and benzylpenicillin, indicating uptake via PEPT2. Accordingly, dipeptide uptake was completely abolished in PEPT2 deficient mice. Reverse transcriptase-polymerase chain reaction analysis detected a PEPT2-specific transcript in extracts from the ganglionic ENS layers of mouse small and large intestine, further proving that enteric dipeptide transport activity is specifically mediated via PEPT2. The cellular site of dipeptide uptake was immunohistochemically localized to enteric glial cells and tissue resident macrophages. In addition, dipeptide uptake occurred in a neurochemically defined subset of neurons in the guinea pig ENS. Our results constitute the first functional evidence for dipeptide transport activity in the ENS. PEPT2-mediated dipeptide transport in enteric glia could contribute to the clearance of neuropeptides in the ENS. In addition, the fluorophore-coupled dipeptide uptake via PEPT2 is a novel vital marker for glial cells in the ENS. PMID- 16041714 TI - Synapse composition and organization following chronic activity blockade in cultured hippocampal neurons. AB - Activity plays multiple roles in the expression of synaptic plasticity, and has been shown to regulate the localization of both neurotransmitter receptors and downstream signaling machinery. However, the role of activity in central synapse formation and organization is incompletely understood. Some studies indicate that synapse formation can occur in the absence of synaptic activity, while others indicate that activity is required for synapse maintenance and receptor recruitment. In addition, the effects of long-term blockade of transmission generally, rather than blockade of specific receptors, on postsynaptic protein complement has been poorly characterized. In order to address the role of activity in synapse formation and postsynaptic specialization, we used tetanus toxin to chronically cleave VAMP2 and inhibit SNARE-mediated neurotransmitter release in cultured hippocampal neurons. Although these neurons are deficient in synaptic release, they are of normal size and morphology. In addition, both excitatory and inhibitory synapses form along their processes with normal density. These synapses have a remarkably similar cellular and molecular organization compared to controls, and are capable of recruiting postsynaptic scaffolding proteins, GABA, and glutamate receptors. Subcellular enrichment of synaptic proteins into specialized domains also appears intact. These data indicate that global activity inhibition is insufficient to disrupt central synapse formation or organization. PMID- 16041715 TI - Developmental modifications of olivocerebellar topography: the granuloprival cerebellum reveals multiple routes from the inferior olive. AB - Correct function of neural circuits depends on highly organized neuronal connections, refined from less precise projections through synaptic elimination, collateral regression, or neuronal death. We examined regressive phenomena that define olivocerebellar topography during maturation from Purkinje cell polyinnervation to monoinnervation. We used bilateral retrograde tracing to determine the source of olivocerebellar afferents to posterior vermis lobules VII VIII in a model of retained immature Purkinje cell polyinnervation, the granuloprival cerebellum. In controls, labelled neurons were found only in the contralateral inferior olive (ION) clustered in a small ventromedial locus that is congruent with known olivocerebellar topography. In granuloprival animals, olivary labelling appeared more dispersed and was present in homologous ipsilateral regions. Double-labelled neurons were never seen. Retrograde tracing following unilateral olivocerebellar transection in adult granuloprival rats revealed: 1) the origin of the normal (remaining) path projecting through the contralateral inferior peduncle was more localized than in irradiated nonpedunculotomized rats, 2) a small double-crossed path, and 3) a projection that ascends the peduncle ipsilateral to the ION of origin, part of which crosses the midline within the cerebellum. Electrophysiological and immunohistochemical assessment in the neonatal cerebellum revealed that transcommissural paths are not present during development but sprout within the irradiated cerebellum. Therefore, the olivocerebellar projection in the granuloprival rat, as a model of the immature path, shows parasagittal organization similar to that of controls in its normally crossed path but possesses additional abnormal projections. Thus, maturation of olivocerebellar topography involves removal of whole developmental paths to define laterality plus synapse elimination within largely predefined parasagittal zones. PMID- 16041716 TI - Development of putative GABAergic neurons in the appendicularian urochordate Oikopleura dioica. AB - Studying the developing brain of urochordates can increase our understanding of brain evolution in the chordate lineage. To begin addressing regional patterns of neuronal differentiation in appendicularian urochordates, we examined the development of putative GABAergic neurons in Oikopleura dioica using GABA immunohistochemistry and in situ hybridization for the GABA-synthesizing enzyme GAD. First, we assessed the developmental dynamics of neuron number and organization in the cerebral and caudal ganglia. We then identified and mapped the positions of putative GABAergic neurons using confocal microscopy. We found GAD mRNA-positive and GABA-immunopositive neurons in the first brain nerves and the cerebral and caudal ganglia, but not in the caudal nerve cord. In both ganglia GAD mRNA-positive and GABA-immunopositive neurons are found in the same characteristic intraganglionic locations. The differentiation of these GABAergic markers occurs first in the first brain nerves and the cerebral ganglion and then with a several-hour delay in the caudal ganglion. In all three structures GAD mRNA expression appears 2-3 hours prior to GABA expression. In general, GABA is expressed by the same number of neurons as express GAD. Several discrepancies suggest differential regulation of the GABAergic phenotype in different neurons, however. Our results show that the GABAergic phenotype has a stereotyped pattern of expression along the anteroposterior axis of the CNS. Given recent genome sequencing and developmental patterning gene studies in this species, the GABAergic neurons in O. dioica provide a good model for assessing, at the invertebrate-vertebrate transition, the molecular mechanisms that specify the GABAergic phenotype. PMID- 16041717 TI - Connexin45 mediates gap junctional coupling of bistratified ganglion cells in the mouse retina. AB - Direction selectivity, a key feature of visual perception, originates in the retina and is transmitted by bistratified ganglion cells that, in the rabbit retina, exhibit a particular coupling pattern. We intracellularly labeled ganglion cells in different transgenic mouse lines, allowing a morphological classification of bistratified ganglion cells, an analysis of their coupling pattern, and the molecular identification of the connexins responsible for the coupling. Based on dendritic characteristics including co-fasciculation with the dendrites of cholinergic starburst amacrine cells, we were able to distinguish three types of bistratified ganglion cells. Two of these co-fasciculate with starburst amacrine cells and exhibit a specific homologous coupling pattern. Connexin45 (Cx45) appears to be the major component of the gap junctional channels because tracer coupling is absent in Cx45-deficient animals whereas it persists in Cx36-deficient animals. It is speculated that the transjunctional voltage dependence of Cx45 channels could support the transmission of direction selectivity. PMID- 16041718 TI - Definition and novel connections of the entopallium in the pigeon (Columba livia). AB - The avian entopallium (E) is the major thalamorecipient zone, within the telencephalon, of the tectofugal visual system. Because of discrepancies concerning the structure of this nuclear mass in pigeons, and in light of recent evidence concerning entopallial projections in other avian species, we here redefine and chart some novel entopallial projections in the pigeon by using a combination of cytochrome oxidase (CO) activity, calcium binding protein immunohistochemistry (CBPi), normal histology, and tract tracing. We show that 1) E is defined by the accurate overlap of CO activity and the dense terminations of thalamic (rotundal) efferents; 2) the perientopallium (Ep), E's overlying belt region, receives a relatively sparse rotundal input and is a major source of projections to wider regions of the hemisphere; and 3) E can be subdivided into internal (Ei) and external (Ex) portions on the basis of normal histology, CBPi, and differential projections. Thus, Ei, but not Ex, makes a reciprocal connection with a distinct nucleus in the ventrolateral mesopallium and is a major source of projections to the lateral striatum. These findings suggest the necessity for a revision of the original proposal of a strictly serial flow of visual information through the entopallial complex and further regions of the hemisphere and also require a modification of the long-standing view that E is comparable to only one specific lamina (IV) of extrastriate visual cortex of mammals. Rather, E appears to be composed of a variety of neuronal types possibly equivalent to those in several neocortical laminae. PMID- 16041719 TI - Novel insect orcokinins: characterization and neuronal distribution in the brains of selected dicondylian insects. AB - Orcokinins are a family of myotropic neuropeptides identified in various decapod crustaceans and recently in a cockroach. Their presence in the crustacean nervous system and hemolymph suggests that they act as hormones and as locally acting neuromodulators. To provide further evidence for the existence of orcokinins in insects, we identified a novel orcokinin-related peptide in the locust Schistocerca gregaria and used an antiserum against Asn13-orcokinin for immunostaining in the brains of selected dicondylian insects, including a silverfish, three polyneopteran species (a cockroach and two locusts), and three endopterygote species (a moth, a bee, and a fly). As analyzed by MALDI-TOF spectrometry and nanoelectrospray Q-TOF, the locust orcokinin is a novel tetradecapeptide with striking sequence similarity to crustacean orcokinins. Orcokinin immunostaining was widespread and occurred in similar patterns in the brain of the silverfish and the polyneopteran species. Prominent immunostaining was detected in the optic lobe, especially in the medulla and in the accessory medulla, in local interneurons of the antennal lobe, and in extrinsic and intrinsic mushroom-body neurons. All parts of the central complex and many other areas of the brains were densely stained. In the silverfish, the cockroach, and the locusts, processes in the corpora cardiaca showed orcokinin immunoreactivity, suggesting that orcokinins also serve a hormonal role. In contrast to the case in polyneopteran species, immunostaining was completely lacking in the brains of the honeybee, fruitfly, and sphinx moth. This indicates that orcokinins either are modified considerably or may be completely absent in the brains of endopterygote insects. PMID- 16041721 TI - Sulcotrione versus atrazine transport and degradation in soil columns. AB - A soil column experiment under outdoor conditions was performed to monitor the fate of 14C-ring-labelled sulcotrione, 2-(2-chloro-4-mesylbenzoyl)cyclohexane-1,3 dione and atrazine, 6-chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine, in water leachates and in the ploughed horizon of a sandy loam soil. Two months after treatment, the cumulative amounts of herbicide residues leached from the soil were 14.5% and 7% of the applied radioactivity for sulcotrione and atrazine, respectively. Maximum leachate concentrations for each herbicide were observed during the first month following application: 120 and 95 microg litre(-1) for sulcotrione and atrazine respectively. After 2 weeks, 78% of the sulcotrione and atrazine was extractable from the soil, whereas after two months only 10 and 4%, respectively, could be extracted. The maximum sulcotrione content in the first 10 cm of soil was identical with that of atrazine. For both molecules, the content of non-extractable residues was low, being around 15%. Sulcotrione seems to be more mobile than atrazine but the consequences for water contamination are similar since lower doses are used. PMID- 16041722 TI - Environmental fate of herbicides trifluralin, metazachlor, metamitron and sulcotrione compared with that of glyphosate, a substitute broad spectrum herbicide for different glyphosate-resistant crops. AB - The introduction of crops resistant to the broad spectrum herbicide glyphosate, N (phosphonomethyl)glycine, may constitute an answer to increased contamination of the environment by herbicides, since it should reduce the total amount of herbicide needed and the number of active ingredients. However, there are few published data comparing the fate of glyphosate in the environment, particularly in soil, with that of substitute herbicides. The objective of this study is to compare the fate of glyphosate in three soils with that of four herbicides frequently used on crops that might be glyphosate resistant: trifluralin, alpha,alpha,alpha-trifluoro-2,6-dinitro-N,N-dipropyl-p-toluidine, and metazachlor, 2-chloro-N-(pyrazol-1-ylmethyl)acet-2',6'-xylidide for oilseed rape, metamitron, 4-amino-4,5-dihydro-3-methyl-6-phenyl-1,2,4-triazin-5-one for sugarbeet and sulcotrione, 2-(2-chloro-4-mesylbenzoyl)cyclohexane-1,3-dione for maize. The distribution of herbicides between the volatilized, mineralized, extractable and non-extractable fractions was studied, along with the formation of their metabolites in laboratory experiments using 14C-labelled herbicides, over a period of 140 days. The main dissipation pathways were mineralization for glyphosate and sulcotrione, volatilization for trifluralin and non-extractable residues formation for metazachlor and metamitron. The five herbicides had low persistence. Glyphosate had the shortest half-life, which varied with soil type, whereas trifluralin had the longest. The half-lives of metazachlor and sulcotrione were comparable, whereas that of metamitron was highly variable. Glyphosate, metazachlor and sulcotrione were degraded into persistent metabolites. Low amounts of trifluralin and metamitron metabolites were observed. At 140 days after herbicide applications, the amounts of glyphosate and its metabolite residues in soils were the lowest in two soils, but not in the third soil, a loamy sand with low pH. The environmental advantage in using glyphosate due to its rapid degradation is counterbalanced by accumulation of aminomethylphosphonic acid specifically in the context of extensive use of glyphosate. PMID- 16041723 TI - Comparative repellency of 38 essential oils against mosquito bites. AB - The mosquito repellent activity of 38 essential oils from plants at three concentrations was screened against the mosquito Aedes aegypti under laboratory conditions using human subjects. On a volunteer's forearm, 0.1 mL of oil was applied per 30 cm2 of exposed skin. When the tested oils were applied at a 10% or 50% concentration, none of them prevented mosquito bites for as long as 2 h, but the undiluted oils of Cymbopogon nardus (citronella), Pogostemon cablin (patchuli), Syzygium aromaticum (clove) and Zanthoxylum limonella (Thai name: makaen) were the most effective and provided 2 h of complete repellency. From these initial results, three concentrations (10%, 50% and undiluted) of citronella, patchouli, clove and makaen were selected for repellency tests against Culex quinquefasciatus and Anopheles dirus. As expected, the undiluted oil showed the highest protection in each case. Clove oil gave the longest duration of 100% repellency (2-4 h) against all three species of mosquito. PMID- 16041724 TI - Antimicrobial activity of some Ganoderma species from Nigeria. AB - The crude n-hexane:diethyl ether, chloroform:acetone and methanol extracts of four species of Ganoderma (Ganoderma colossum (Fr.) C. F. Baker, G. resinaceum Boud., G. lucidum (cf.) (Curtis) P. Karst. and G. boninense (cf.) Pat.), from Nigeria, were tested for antimicrobial activity. The three solvent extracts of all the species of Ganoderma were active against Pseudomonas syringae and Bacillus subtilis, whereas none of the extracts were active against Cladosporium herbarum. Preliminary thin layer chromatography chemical tests on these extracts of Ganoderma showed that they contained compounds that stained blue-violet and blue or green when sprayed with anisaldehyde-sulphuric acid or Dragendorff, respectively. The profile of compounds in the extracts showed some variation among the four species. PMID- 16041725 TI - Aged garlic extract improves endothelial function in men with coronary artery disease. AB - Endothelium-dependent vasodilation is impaired and predicts the risk of a coronary event in patients with coronary artery disease (CAD). Oxidant stress and increased systemic inflammation may contribute to this endothelial dysfunction. Aged garlic extract (AGE) contains antioxidant compounds and increases nitric oxide production and decreases the output of inflammatory cytokines from cultured cells. The aim of this study was to test the effect of treatment with AGE on brachial artery flow mediated endothelium-dependent dilation (FMD) and circulating markers of oxidative stress and systemic inflammation. The trial included 15 men with angiographically proven CAD in a randomized, placebo controlled, cross-over design with 2-week treatment and washout periods. During AGE supplementation, FMD increased (44%) significantly (p = 0.04) from the baseline and mainly in men with lower baseline FMD. Levels of FMD at the end of AGE treatment were significantly (p = 0.03) higher compared with the corresponding levels at the end of placebo treatment when the variation in baseline body weight was taken into account. Markers of oxidant stress (plasma oxidized low density lipoprotein and peroxides), systemic inflammation (plasma C reactive protein ad interleukin-6) and endothelial activation (VCAM-1) did not change significantly during the study. These data suggest that short-term treatment with AGE may improve impaired endothelial function in men with CAD treated with aspirin and a statin. Whether improvement in endothelial function decreases the risk of future cardiovascular events remains to be determined. PMID- 16041726 TI - Secondary metabolites from Chamaedora tepejilote (Palmae) are active against Mycobacterium tuberculosis. AB - Due to the emergence of multi-drug resistant Mycobacterium tuberculosis strains, there is an urgent need to search for new antimycobacterial drugs. The present work describes the bioactivity-guided fractionation of Chamaedora tepejilote hexane extract which led to the isolation and the characterization of squalene, farnesol, methylic ester of hexadecanoic acid, beta-sitosterol and ursolic acid. Activity against Mycobacterium tuberculosis H37Rv, determined by radiorespirometric Bactec assay, showed ursolic acid, squalene and farnesol to produce a M. tuberculosis growth inhibition of 99% at a concentration of 100 microg/mL. PMID- 16041727 TI - Comparison of the antiinflammatory effects of Drosera rotundifolia and Drosera madagascariensis in the HET-CAM assay. AB - The antiinflammatory effects of ethanol and aqueous extracts from Drosera rotundifolia and from Drosera madagascariensis were compared in vivo in the HET CAM assay. Both extracts from D. rotundifolia and the ethanol extract from D. madagascariensis showed remarkable efficacy at doses of 500 microg/pellet. The inhibition of the inflammation by the extracts was stronger than that by 50 microg hydrocortisone/pellet. In contrast, there was only a very weak effect observed at a dose of 500 microg/pellet of the water extract from D. madagascariensis. The chemical analyses of the extracts showed that the effect cannot be attributed to naphthoquinones, but might be due to flavonoids. Ellagic acid obviously plays an important role in the antiangiogenic effect of the Drosera extracts. PMID- 16041728 TI - Garlic natural health products exhibit variable constituent levels and antimicrobial activity against Neisseria gonorrhoeae, Staphylococcus aureus and Enterococcus faecalis. AB - The composition of 19 garlic natural health products (NHPs) and fresh garlic extracts were determined, as was their antibacterial activity. The 19 NHPs and 5 fresh garlic extract standards were analysed for their principal active constituents. They were also extracted for 5, 10 or 15 min in water to fresh garlic equivalents of 200 mg/mL. The extract's minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) against three indicator microorganisms (Neisseria gonorrhoeae, Staphylococcus aureus and Enterococcus faecalis) were determined by the broth microdilution method. While 47% of the aqueous garlic NHP extracts exhibited activity against N. gonorrhoeae, only 16% of the aqueous extracts inhibited S. aureus or E. faecalis at all three timepoints. Generally, products with high antimicrobial activity contained higher levels of garlic constituents with comparable activity to fresh garlic extracts, while products with marginal antibacterial activity often contained lower concentrations of constituents than their product labels indicated. Different extraction times affected antibacterial activity only against N. gonorrhoeae and tended to be correlated with levels of allicin. Thus, many extracts showed discrepancies in both composition, allicin:alliin ratio and antimicrobial activity, raising concerns as to standards of preparation and quality control for these products. PMID- 16041729 TI - Site-specific detection of S-nitrosylated PKB alpha/Akt1 from rat soleus muscle using CapLC-Q-TOF(micro) mass spectrometry. AB - Protein Kinase Balpha(PKBalpha, or Akt1) is believed to play a crucial role in programmed cell death, cancer progression and the insulin-signaling cascade. The protein is activated by phosphorylation at multiple sites and subsequently phosphorylates and activates eNOS. Free cysteine residues of the protein may capture reactive, endogenously produced nitric oxide (NO) as S-nitrosothiols. Site-specific detection of S-nitrosylated cysteine residues, usually at low stoichiometry, has been a major challenge in proteomic research largely due to the lack of mass marker for S-nitrosothiols that are very labile under physiologic conditions. In this report we describe a sensitive and specific MS method for detection of S-nitrosothiols in PKB alpha/Akt1 in rat soleus muscle. PKB alpha/Akt1 was isolated by immunoprecipitation and 2D-gel electrophoresis, subjected to in-gel tryptic digestion, and cysteinyl nitrosothiols were reacted with iodoacetic acids [2-C(12)/C(13) = 50/50] under ascorbate reduction conditions. This resulted in the production of relatively stable carboxymethylcysteine (CMC) immonium ions (m/z 134.019 and m/z 135.019) within a narrow argon collision energy (CE = 30 +/- 5 V) in the high MS noise region. In addition, free and disulfide-linked cysteine residues were converted to carboxyamidomethylcysteines (CAM). Tryptic S-nitrosylated parent ion was detected with a mass accuracy of 50 mDa for the two CMC immonium ions at the triggered elution time during capillary liquid chromatography (LC) separation. A peptide containing Cys(296) was discriminated from four co-eluting tryptic peptides under lock mass conditions (m/z 785.8426). S-nitrosothiol in the tryptic peptide, ITDFGLBKEGIK (B: CAM, [M + 2H](2+) = 690.86, Found: 690.83), is believed to be present at a very low level, since the threshold for the CMC immonium trigger ions was set at 3 counts/s in the MS survey. The high levels of NO that are produced under stress conditions may result in increased S-nitrosylation of Cys(296) which blocks disulfide bond formation between Cys(296) and Cys(310) and suppresses the biological effects of PKB alpha/Akt1. With the procedures developed here, this process can be studied under physiological and pathological conditions. PMID- 16041730 TI - Optimized sample preparation for isotopic analyses of CO2 in air: systematic study of precision and accuracy dependence on driving variables during CO2 purification process. AB - A systematic analysis of efficiency, reproducibility and accuracy of cryogenic purification of CO(2) from air samples for isotopic analyses is presented. The technical characteristics of the cryogenic line are given in detail. To study the cryogenic process, three different operating parameters are considered: flow rate of the gas entering the line, pressure of the gas in the line, and CO(2)-trap shape. Experimental results demonstrate that efficiency, reproducibility and accuracy strongly depend on the CO(2)trap shape. Moreover, a dependence of reproducibility and accuracy on the flow rate of the gas is found, but not on its pressure. High precision (< or =0.02 per thousand for delta(13)C and < or =0.05 per thousand for delta(18)O) and good accuracy (<0.09 per thousand for delta(13)C and <0.14 per thousand for delta(18)O) is achieved after applying the N(2)O correction. PMID- 16041731 TI - A randomized double-blind placebo-controlled trial of a standardized extract of Matricariae recutita, Foeniculum vulgare and Melissa officinalis (ColiMil) in the treatment of breastfed colicky infants. AB - OBJECTIVE: The aim of this randomized, double-blind, placebo-controlled trial was to investigate the effectiveness and side effects of a phytotherapeutic agent with Matricariae recutita, Foeniculum vulgare and Melissa officinalis in the treatment of infantile colic. METHODS: 93 breastfed colicky infants were enrolled, the diagnosis was made according to Wessel's criteria. After a 3 day observation period, the infants were randomly divided into two groups, one treated with phytotherapeutic agent (PA) and the other with placebo twice a day for 1 week. Crying time and side effects were recorded. RESULTS: 88 infants completed the trial: 41 in the PA group and 47 in the control. The daily average crying time for the PA was 201.2 min/day (SD 18.3) at the baseline and 76.9 min/day (SD 23.5) at the end of the study; for the placebo it was 198.7 min/day (SD 16.9) and 169.9 min/day (SD 23.1) (p < 0.005). Crying time reduction was observed in 85.4% subjects for the PA and in 48.9% subjects for the placebo (p < 0.005). No side effects were reported. CONCLUSION: The present study shows that colic in breastfed infant improves within 1 week of treatment with an extract based on Matricariae recutita, Foeniculum vulgare and Melissa officinalis. PMID- 16041732 TI - Effect of pumpkin seed (Cucurbita pepo) protein isolate on the activity levels of certain plasma enzymes in CCl4-induced liver injury in low-protein fed rats. AB - The effects of pumpkin seed (Cucurbita pepo) protein isolate on the activity levels of lactate dehydrogenase (LD), alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) against carbon tetrachloride (CCl4)-induced acute liver injury in low-protein fed rats were investigated. A group of male Sprague-Dawley rats maintained on a low-protein diet for 5 days were divided into three subgroups. Two subgroups were injected with carbon tetrachloride and the other group with an equivalent amount of olive oil. Two hours after CCl4 intoxication one of the two subgroups was administered with pumpkin seed protein isolate. All three subgroups of rats were maintained on the low-protein diet for the duration of the investigation. Groups of rats from the different subgroups were killed at 24, 48 and 72 h after their respective treatments. After 5 days on the low-protein diet the activity levels of all four enzymes were significantly higher than their counterparts on a normal balanced diet. CCl4 intoxication resulted in significant increases in the activity levels of all four enzymes investigated. The administration of pumpkin seed protein isolate after CCl4 intoxication resulted in significantly reduced activity levels of all four enzymes. It is concluded that pumpkin seed protein isolate administration was effective in alleviating the detrimental effects associated with protein malnutrition. PMID- 16041733 TI - Stimulation of lymphocyte proliferation and inhibition of nitric oxide production by aqueous Urtica dioica extract. AB - The immunomodulatory and antiinflammatory activities of aqueous Urtica dioica extract were investigated for their effect on the mitogenic response of murine splenocytes and nitric oxide production by murine peritoneal macrophages in vitro. It was found that this extract stimulated the proliferation of T lymphocytes and suppressed NO production in lipopolysaccharide-stimulated macrophages without affecting cell viability. PMID- 16041734 TI - Clinical evaluation of the traditional chinese prescription Chi-Ju-Di-Huang-Wan for dry eye. AB - BACKGROUND: Dry eye is a very common ocular disease characterized by eye dryness and photophobia. It often influences the patient's normal life. This study evaluated the therapeutic effects of the Chinese herb, Chi-Ju-Di-Huang-Wan, for treatment of dry eye patients. It is one of the most commonly prescribed Chinese herbs used for eyes, however, there are no scientific reports about its effect. METHODS: This study included 80 dry eye patients. The patients were randomly divided into two groups. The experimental group was treated with topical eye drops and Chi-Ju-Di-Huan-Wan. The control group was given topical eye drops and placebo. All patients received tear film examinations including Schirmer's test, Rose Bengal test, fluorescein stain test, break up time test and questionnaires. RESULTS: The results of the tear film tests were analysed by two independent t tests between the two groups. No significant differences between the two groups according to the Schirmer's test were found. There was a significant difference in the Rose Bengal test at week 2 and tear break up time at week 4 (p = 0.04, 0.04, individually). CONCLUSION: Chi-Ju-Di-Huang-Wan is an effective stabilizer of tear film and decreases the abnormality of corneal epithelium. It provides an alternative choice for dry eye treatment. PMID- 16041735 TI - Inhibition of hepatitis B virus by an aqueous extract of Agrimonia eupatoria L. AB - Inhibition of HBsAg release against hepatitis B virus (HBV) was investigated in an aqueous extract prepared from the aerial parts (stems and leaves) of Agrimonia eupatoria. The inhibitory effect on HBsAg secretion was footed using aqueous extracts of Agrimonia eupatoria at four different temperatures (37 degrees C 45 degrees C, 55 degrees C and 60 degrees C), and the extract prepared at 60 degrees C was found to have the greatest effect. The inhibitory activity of Agrimonia eupatoria extracts on HBsAg secretion varied over the growing season and was the highest at mid-July. This inhibitory activity was also shown with the aqueous extracts of two other species of the genus Agrimonia: A. pilosa and A. coreana pilosella. These results suggest that some plants of the genus Agrimonia contain potential antiviral activity against HBV. PMID- 16041736 TI - Avilamycin did not play a role in the discontinuation of evernimicin as a clinical drug candidate. PMID- 16041737 TI - Screening of Korean medicinal plants for lipase inhibitory activity. AB - The pancreatic lipase inhibitory activity of the aqueous ethanol extracts obtained from 19 medicinal plants was evaluated in vitro by a continuous monitoring pH-Stat technique using tributyrin as a substrate. Of the extracts tested, those of Juniperus communis (bark) and Illicium religiosum (wood) exhibited the strongest activity with an IC50 value of 20.4 and 21.9 microg/mL, respectively. PMID- 16041738 TI - Essential oil composition of Piper guineense and its antimicrobial activity. Another chemotype from Nigeria. AB - The analysis of the essential oil of Piper guineense from Nigeria presents a new chemotype of constituents different from earlier reports with the absence of the usual myristicin. Ishwarane, a common constituent of Aristolochia indica and Bixa orellana, was also isolated from the essential oil of the fruit. The essential oil inhibited the growth of Pseudomonas aeruginosa UCH 655 strain at 5 mg/mL on which standard antibiotic drugs were ineffective. PMID- 16041739 TI - Efficacy and tolerance of atrisin in degenerative and inflammatory joint disorders. AB - In a randomized multi-centre clinical trial the efficacy and tolerance of a herbal product Atrisin three capsules/day was evaluated in 65 patients (31 male, 34 female) suffering from osteoarthritis and rheumatoid arthritis, during a period of 2 months. Assessment of pain and functional disability were made on a 10 cm horizontal visual analogue scale. The severity of osteoarthritis was evaluated by Lequesne's index.Spontaneous pain showed significant improvement. Similarly there was progressive and significant reduction in the Lequesne's functional index. At the completion of the study patients taking Atrisin were using less NSAIDs. Atrisin was tolerated well by the patients and there were no adverse cardiovascular or gastric effects reported. There was feeling of general well-being and compliance by the patients. PMID- 16041740 TI - Fragmentation reactions of deprotonated peptides containing proline. The proline effect. AB - The collision-induced dissociation (CID) fragmentation reactions of a variety of deprotonated peptides containing proline have been studied in detail using MS(2) and MS(3) experiments, deuterium labelling and accurate mass measurements when necessary. The [M--H--CO(2)](-) (a(2)) ion derived from H-Pro-Xxx-OH dipeptides shows an unusual fragmentation involving loss of C(2)H(4); this fragmentation reaction is not observed for larger peptides. The primary fragmentation reactions of deprotonated tripeptides with an N-terminal proline are formation of a(3) and y(1) ions. When proline is in the central position of tripeptides, a(3), y(2) and y(1) ions are the primary fragmentation products of [M--H](-), while when the proline is in the C-terminal position, a(3)and y(1) ions are the major primary products. In the latter case, the a(3) ion fragments primarily to the ''b(2) ion; further evidence is presented that the ''b(2) ions have a deprotonated oxazolone structure. Larger deprotonated peptides having at least two amino acid residues N terminal to proline show a distinct preference for cleavage of the amide bond N terminal to proline to form, mainly, the appropriate y ion. This proline effect is compared and contrasted with the similar proline effect observed in the fragmentation of protonated peptides containing proline. PMID- 16041741 TI - Morphing into cancer: the role of developmental signaling pathways in brain tumor formation. AB - Morphogens play a critical role in most aspects of development, including expansion and patterning of the central nervous system. Activating germline mutations in components of the Hedgehog and Wnt pathways have provided evidence for the important roles morphogens play in the genesis of brain tumors such as cerebellar medulloblastoma. In addition, aberrant expression of transforming growth factor-beta (TGF-beta) superfamily members has been demonstrated to contribute to progression of malignant gliomas. This review summarizes our current knowledge about the roles of morphogens in central nervous system tumorigenesis. PMID- 16041742 TI - Brain as a paradigm of organ growth: Hedgehog-Gli signaling in neural stem cells and brain tumors. AB - The Hedgehog-Gli (Hh-Gli) signaling pathway is essential for numerous events during the development of many animal cell types and organs. In particular, it controls neural cell precursor proliferation in dorsal brain structures and regulates the number of neural stem cells in distinct embryonic, perinatal, and adult niches, such as the developing neocortex, the subventricular zone of the lateral ventricle of the forebrain, and the hippocampus. We have proposed that Hh Gli signaling regulates dorsal brain growth during ontogeny and that its differential regulation underlays evolutionary change in the morphology (size and shape) of dorsal brain structures. It is also critically involved in sporadic brain tumorigenesis--as well as several other human cancer--suggesting that tumors derive from stem cells or progenitors maintaining an inappropriate active Hh-Gli pathway. Importantly, we and others have demonstrated that human sporadic tumors from the brain and other organs require sustained HH-GLI signaling for sustained growth and survival. Modulating HH-GLI signaling thus represents a novel rational avenue to treat, on one hand, brain degeneration and injury by inducing controlled HH-GLI-mediated regeneration and growth, and on the other hand, to combat cancer by blocking its abnormal activity in tumor cells. PMID- 16041743 TI - New concepts for regio- and stereoselective bis- and triscyclopropanations of C60. AB - The synthesis of isomerically multiple adducts of C(60) with a defined three dimensional structure is still one of the most challenging tasks of exohedral fullerene chemistry. The inherent regioselectivity of successive additions of addends such as malonates to the fullerene's [6,6]-double bonds is only moderate. In most cases difficult-to-isolate mixtures of regioisomers are obtained. The regioselectivity can be significantly improved if multifunctional addends able to undergo two or more additions are allowed to react with C(60). Preorganization and minimization of strain energy within the addend skeleton reduce the number of sterically allowed addition patterns. Improved concepts for highly regio- and stereoselective bis- and triscyclopropanations of C(60) are described. Two examples of the bisadditions with complete regioselectivity leading to trans-2- and cis-2 are presented. Here, the two malonate binding sites are linked by rigid tetraphenylporphyrin and calix-[4]-arene spacers. Selective trisadditions were achieved with the easy-to-synthesize and easy-to-modify tripodal addends 5-7, where the malonates are held together by a focal aryl moiety. Another very elegant approach for bis- and trisadditions involves cyclo-[n]-alkylmalonates. Selection between addition patterns with and without rotational axes is possible by choosing the right combinations of the flexible alkyl chains connecting the malonates. If alkyl chains of identical lengths are used bis- and trisadducts such as 19-21 and 25 with rotational symmetry are formed with high regioselectivity. These addition patterns are avoided if cyclo-[n]-malonates containing alkyl chains of different lengths are employed. In this case adducts such as 26 and 27 with C(s)-symmetry are formed. The use of the chiral cyclo-[3] malonate 28 allows for the regio- and stereoselective synthesis of the enantiomerically pure e,e,e-trisadducts 29 and 30 containing an inherently chiral addition pattern with C(3)-symmetry. PMID- 16041744 TI - Enzyme inhibitors as chemical tools to study enzyme catalysis: rational design, synthesis, and applications. AB - Carefully designed molecules that are intimately related to the reaction mechanism of enzymes are often highly selective and potent inhibitors that serve as extremely useful chemical probes for understanding the reaction mechanism and structure of enzymes. This article describes the design, synthesis, and applications of specific inhibitors of two mechanistically distinct groups of enzymes, ATP-dependent amide ligases and Ser- and Thr-hydrolases. Our strategy is based on the premise that stable analogues of the transition state (transition state analogues) are highly potent inhibitors that serve as good mechanistic probes, and that a key structure of a good inhibitor of one enzyme is also utilized for the inhibitors of other enzymes that share the same chemistry in their catalyzed reactions, irrespective of the degree of structural similarity and evolutionary link between the enzymes. According to these principles, we designed and synthesized a series of phosphinate- and sulfoximine-based transition-state analogue inhibitors of glutathione synthetase, gamma glutamylcysteine synthetase and asparagine synthetase. For the second group of enzymes, we synthesized a gamma-monofluorophosphono glutamate analogue for mechanism-based affinity labeling of gamma-glutamyltranspeptidase and fluorescent phosphonic acid esters for the active-site titration of lipase. These inhibitors were used successfully as ligands for detailed kinetic analyses, X-ray crystallography, and mass analysis of the enzymes to identify the key amino acid residues responsible for catalysis and substrate recognition in the transition state. PMID- 16041745 TI - Toward the development of new medicinal leads with selectivity for protein kinase C isozymes. AB - Tumor promoters such as phorbol esters bind strongly to protein kinase C (PKC) isozymes to induce their activation. Since each PKC isozyme is involved in diverse biological events in addition to tumor promotion, the isozymes serve as promising therapeutic targets. Tumor promoters bind to the C1A and/or C1B domain of conventional (alpha, betaI, betaII, and gamma) and novel PKC isozymes (delta, epsilon, eta, and theta). As these C1 domains play differential roles in PKC activation and their translocation in cells, the development of agents with binding selectivity for individual C1 domains is a pressing need. For this purpose, we established a synthetic C1 peptide library of all PKC isozymes. The library enabled us to identify indolactam-V (1) as a promising lead compound. Our diverse structure-activity studies on 1 indicated that the position of the hydrophobic substituent on the indole ring dominates the PKC isozyme- and C1 domain-selective binding rather than conformation of the nine-membered lactam. Moreover, we suggested that the indole ring of 1 could be involved in the CH/pi interaction with Pro-11 of the C1B domain of PKCdelta. This invaluable information will lead to the structural optimization of the PKCdelta ligand as exemplified by the design and synthesis of naphtholactam-V8 (21). PMID- 16041746 TI - Natural products as tools for studies of ligand-gated ion channels. AB - Ligand-gated ion channels, or ionotropic receptors, constitute a group of membrane-bound proteins that regulate the flux of ions across the cell membrane. In the brain, ligand-gated ion channels mediate fast neurotransmission. They are crucial for normal brain function and involved in many diseases in the brain. Historically, natural products have been used extensively in biomedical studies and ultimately as drugs or leads for drug design. In studies of ligand-gated ion channels, natural products have been essential for the understanding of their structure and function. In the following a short survey of natural products and their use in studies of ligand-gated ion channels is given. PMID- 16041747 TI - Anatomy in basic surgical training. PMID- 16041748 TI - Reduced undergraduate medical science teaching is detrimental for basic surgical training. PMID- 16041749 TI - Medical science teaching needs stronger support from universities and a better career structure for anatomists. PMID- 16041750 TI - Changing times, changing training: anatomy teaching in basic surgical training. PMID- 16041751 TI - Introduction: unexpected roles for morphogens in the development and regeneration of the CNS. PMID- 16041752 TI - Morphogens and cell survival during development. AB - The notion of "morphogens" is an important one in developmental biology. By definition, a morphogen is a molecule that emanates from a specific set of cells that is present in a concentration gradient and that specifies the fate of each cell along this gradient. The strongest candidate morphogens are members of the transforming growth factor-beta (TGF-beta), Hedgehog (Hh), and Wnt families. While these morphogens have been extensively described as differentiation inducers, some reports also suggest their possible involvement in cell death and cell survival. It is frequently speculated that the cell death induction that is found associated with experimental removal of morphogens is the manifestation of abnormal differentiation signals. However, several recent reports have raised controversy about this death by default, suggesting that cell death regulation is an active process for shaping tissues and organs. In this review, we will present morphogens, with a specific emphasis on Sonic Hedgehog, a mammalian member of the Hh family, not as a positive regulators of cell differentiation but as key regulators of cell survival. PMID- 16041753 TI - Role of morphogens in brain growth. AB - During the last century, experiments on the chick embryo established that the ballooning expansion of the early forebrain and midbrain vesicles is dependent on the underlying axial (notochordal) mesoderm. Transient separation of the early midbrain primordium from the notochord causes subsequent collapse of both midbrain and forebrain (telencephalic) vesicles, accompanied by pronounced folding of the neural epithelium. More recent experiments have shown that vesicle collapse is caused by defective Sonic Hedgehog (Shh) signaling from the notochord and floor plate. Separation of the notochord from the brain causes loss of ventral Shh expression, resulting in reduced cell proliferation and increased cell death in the expanding neural epithelium, and culminating in vesicle collapse. These experiments are reviewed here, and set in the context of other studies illustrating the wide range of molecular and cellular processes that cause abnormal brain morphogenesis when perturbed. We also speculate that variation in the regulation of signaling pathways such as Hedgehog may have played a significant part in generating rapid morphogenetic changes during the evolution of the vertebrate brain. PMID- 16041754 TI - Morphogens in motion: growth control of the neural tube. AB - The entire vertebrate nervous system develops from a simple epithelial sheet, the neural plate which, along development, acquires the large number and wide variety of neuronal cell types required for the construction of a functional mature nervous system. These include processes of growth and pattern formation of the neural tube that are achieved through complicated and tightly regulated genetic interactions. Pattern formation, particularly in the vertebrate central nervous system, is one of the best examples of a morphogen-type of function. Cell cycle progression, however, is generally accepted to be dependent on cell-intrinsic factors. Recent studies have demonstrated that proliferation of neural precursors is also somehow controlled by secreted signaling molecules, well-known by their role as morphogens, such as fibroblast growth factor (FGF), vertebrate orthologs of the Drosophila wingless (Wnt), hedgehog (Hh), and transforming growth factor beta (TGF-beta) families, that in turn regulate the activity of factors controlling cell cycle progression. In this review we will summarize the experimental data that support the idea that classical morphogens can be reused to regulate proliferation of neural precursors. PMID- 16041755 TI - Morphogen signaling at the vertebrate growth cone: a few cases or a general strategy? AB - Axon navigation relies on the competence of growth cones to sense and interpret attractive and repulsive guidance cues present along their trajectory. For most neurons, this process is mediated by a limited number of conserved families of ligand-receptor signaling systems, including Ephrin/Eph, Netrins/DCC-Unc5, Slits/Robo, and Semaphorins/Plexin-Neuropilin. Recent studies have demonstrated that some neurons respond also to well-known secreted signaling molecules, best known for their roles as morphogens, such as BMP7, SHH, FGF8, and Wnt. Thus, retina ganglion cell axon navigation is influenced by FGF, SHH, and possibly BMP signaling. Similarly, commissural neurons in the spinal cord respond sequentially to the activity of BMP, SHH, and Wnt to extend toward and away from their intermediate target, the floor plate. The data that support this conclusion will be summarized and how morphogens may signal at the growth cone will be discussed. PMID- 16041756 TI - Morphogens and synaptogenesis in Drosophila. AB - During development and adult life synapses are remodeled in response to genetic programs and environmental cues. This synaptic plasticity is thought to be the basis of learning and memory. The larval neuromuscular junction of Drosophila is established during embryogenesis and grows during larval development to accommodate muscle growth and maintain synaptic homeostasis. This growth is dependent on bidirectional communication between the motoneuron and the muscle fiber. The best-characterized retrograde signaling pathway is defined by Glass bottom boat (Gbb), a morphogen of the transforming growth factor-beta (TGF-beta) superfamily. Gbb acts as a muscle-derived retrograde signal that activates the TGF-beta pathway presynaptically. This pathway includes the type II receptor Wishful thinking, type I receptors Thick veins and Saxophone, and the second messenger Smads Mothers against dpp (Mad) and Medea. Mutations that block this pathway result in small synapses that are morphologically aberrant and severely impaired functionally. An emerging anterograde signaling pathway is defined by Wingless, a morphogen of the Wnt family that acts as a motoneuron-derived anterograde signal required for both pre- and postsynaptic development. In the absence of Wingless the neuronal microtubule cytoskeleton regulator Futsch is down-regulated and synaptic growth impaired. Some of these morphogens have conserved roles in mammalian synaptogenesis, and genetic analysis suggests that additional signaling molecules are required for synaptic growth at the Drosophila neuromuscular junction. PMID- 16041757 TI - CNS regeneration: a morphogen's tale. AB - Tissue regeneration will soon become an avenue for repair of damaged or diseased tissues as stem cell niches have been found in almost every organ of the vertebrate body including the CNS. In addition, different animals display an array of regenerative capabilities that are currently being researched to dissect the molecular mechanisms involved. This review concentrates on the different ways in which CNS tissues such as brain, spinal cord and retina can regenerate or display neurogenic potential and how these abilities are modulated by morphogens. PMID- 16041758 TI - Mechanisms of morphogen movement. AB - Morphogens are defined as signaling molecules that are produced locally, yet act directly at a distance to pattern the surrounding field of cells in a concentration-dependent manner. In recent years many laboratories have devoted their attention to how morphogens actually reach distant cells. Several models have been proposed, including diffusion in the extracellular space and planar transcytosis. A combination of genetic, developmental, and cell-biological approaches have been taken to tackle this issue. I will present the models and discuss the types of experiments that have been designed to test them. It stands out that most of the work has been carried out in Drosophila. Morphogens contribute to patterning of the vertebrate nervous system, and the same signaling molecules have recently been shown to play important, possibly instructive, roles in axon guidance. Little, if anything, is known about the movement of morphogens in the context of nervous system development. The long-standing tradition of biophysical studies on diffusion in the brain extracellular space, along with the sophisticated in vitro culture systems developed in neurobiology laboratories, may provide new tools and ideas to test these models in a new context. PMID- 16041759 TI - Mechanisms of Hedgehog gradient formation and interpretation. AB - Morphogens are molecules that spread from localized sites of production, specifying distinct cell outcomes at different concentrations. Members of the Hedgehog (Hh) family of signaling molecules act as morphogens in different developmental systems. If we are to understand how Hh elicits multiple responses in a temporally and spatially specific manner, the molecular mechanism of Hh gradient formation needs to be established. Moreover, understanding the mechanisms of Hh signaling is a central issue in biology, not only because of the role of Hh in morphogenesis, but also because of its involvement in a wide range of human diseases. Here, we review the mechanisms affecting the dynamics of Hh gradient formation, mostly in the context of Drosophila wing development, although parallel findings in vertebrate systems are also discussed. PMID- 16041760 TI - Role of morphogens in neural crest cell determination. AB - The neural crest is a transient, migratory cell population found in all vertebrate embryos that generate a diverse range of cell and tissue derivatives including, but not limited, to the neurons and glia of the peripheral nervous system, smooth muscle, connective tissue, melanocytes, craniofacial cartilage, and bone. Over the past few years, many studies have provided tremendous insights into understanding the mechanisms regulating the induction and migration of neural crest cell development. This review highlights the surprising and perhaps unexpected roles for morphogens in these distinct processes. A comparison of studies performed in several different vertebrates emphasizes the requirement for coordination between multiple signaling pathways in the induction and migration of neural crest cells in the developing embryo. PMID- 16041761 TI - Signaling at the vertebrate synapse: new roles for embryonic morphogens? AB - The formation of synapses is critical for functional neuronal connectivity. The coordinated assembly at both sides of the synapse is fundamental for the proper apposition of the neurotransmitter release machinery on the presynaptic neuron and the clustering of neurotransmitter receptors and ion channels on the receptive postsynaptic cell. This process requires bidirectional communication between the presynaptic neuron and its postsynaptic target, another neuron, or muscle fiber. Extracellular signals such as WNT, TGF-beta, and FGF factors are emerging as key target-derived signals required for the initial stages of synaptic assembly. Studies in invertebrates are also providing new insights into the function of these signals in synaptic growth and homeostasis. During early embryonic patterning, WNT, TGF-beta, and FGF factors function as typical morphogens in a concentration-dependent manner to regulate cell fate decisions. This mode of action raises the provocative idea that these same morphogens might also provide a coordinate system for axons to establish the distance to their targets during axon guidance and synapse formation. PMID- 16041762 TI - Planar cell polarity and a potential role for a Wnt morphogen gradient in stereociliary bundle orientation in the mammalian inner ear. AB - The planar cell polarity (PCP) pathway, a noncanonical Wnt signaling pathway, is crucial for embryonic development in all animals as it is responsible for the regulation of coordinated orientation of structures within the plane of the various epithelia. In the mammalian cochlea, one of the best examples of planar polarity in vertebrates, stereociliary bundles located on mechanosensory hair cells within the sensory epithelium are all uniformly polarized. Generation of this polarity is important for hair cell mechanotransduction and auditory perception as stereociliary bundles are only sensitive to vibrations in their single plane of polarization. We describe the two step developmental process that results in the generation of planar polarity in the mammalian inner ear. Furthermore, we review evidence for the role of Wnt signaling, and the possible generation of a Wnt gradient, in planar polarity. PMID- 16041763 TI - New designer drug 2,5-dimethoxy-4-ethylthio-beta-phenethylamine (2C-T-2): studies on its metabolism and toxicological detection in rat urine using gas chromatography/mass spectrometry. AB - Studies are described on the metabolism and the toxicological analysis of the phenethylamine-derived designer drug 2,5-dimethoxy-4-ethylthio-beta phenethylamine (2C-T-2) in rat urine using gas chromatography/mass spectrometry (GC/MS) after enzymatic cleavage of conjugates, liquid-liquid extraction and derivatization. The structures of 14 metabolites were assigned tentatively by detailed interpretation of their mass spectra. Identification of these metabolites indicated that 2C-T-2 was metabolized by sulfoxidation followed by N acetylation and either hydroxylation of the S-ethyl side chain or demethylation of one methoxy group, O-demethylation of the parent compound followed by N acetylation and sulfoxidation, deamination followed by reduction to the corresponding alcohol followed by partial glucuronidation and/or sulfation or by oxidation to the corresponding acid followed either by partial glucuronidation or by degradation to the corresponding benzoic acid derivative followed by partial glucuronidation. Furthermore, 2C-T-2 was metabolized by N-acetylation of the parent compound followed either by O-demethylation and sulfoxidation or by S dealkylation, S-methylation and sulfoxidation. The authors' systematic toxicological analysis (STA) procedure using full-scan GC/MS after acid hydrolysis, liquid-liquid extraction microwave-assisted acetylation allowed the detection of an intake of a dose of 2C-T-2 in rat urine, which corresponds to a common drug users' dose. Assuming similar metabolism, the described STA procedure should be suitable for proof of an intake of 2C-T-2 in human urine. PMID- 16041764 TI - Lipoxygenase inhibitors from natural plant sources. Part 2: medicinal plants with inhibitory activity on arachidonate 12-lipoxygenase, 15-lipoxygenase and leukotriene receptor antagonists. AB - The metabolism of arachidonic acid can be catalysed by either one of two enzyme families: the cyclooxygenases or the lipoxygenases. The lipoxygenase enzymes are classed into several subcategories including 5-, 12- and 15-lipoxygenases. The 5 lipoxygenase pathway has been the major focus of study due to the pronounced pro inflammatory role of leukotrienes and the approval of 5-lipoxygenase inhibitors and leukotriene receptor antagonists for the clinical treatment of asthma. Although less well characterized, the 12-lipoxygenase as well as the 15 lipoxygenase pathway may also play an important role in the progression of human diseases such as cancer, psoriasis and atherosclerosis. The present review article summarizes the findings from an extensive literature search on plants that have been assessed for 12- and 15-lipoxygenase inhibitory activity as well as for leukotriene receptor antagonistic properties. The results are presented in a tabular format, and a discussion about promising plant species and natural compounds as well as relevant in vitro assays are included in this article. PMID- 16041765 TI - Flavonoids from the leaves of Litsea japonica and their anti-complement activity. AB - Four flavonoids, epicatechin (1), afzelin (2), quercitrin (3), and tiliroside (4), were isolated from the leaves of Litsea japonica (Thunb.) Jussieu (Lauraceae). The structures of compounds were identified by comparing their chemical and spectral data with those previously reported. The flavonoids (1-4) were tested for their anti-complement activity against classical pathway of complement system. Compounds 2-4 showed inhibitory activity against complement system with IC50 values of 258, 440, and 101 microm, respectively, whereas 1 was inactive. For the evaluation of the structure-activity relationship of 5,7 dihydroxyflavones, myricitrin (5) from Juglans mandshurica also tested for it's anti-complement activity and is inactive in this assay system. Furthermore, compounds 2, 3, and 5 were hydrolyzed with naringinase to give kaempferol (2a), quercetin (3a), and myricetin (5a), and these were also tested for their activity. Of the three aglycones, 2a exhibited anti-complement activity with an IC50 value of 730 microM, while 3a and 5a were inactive. The inhibitory potencies of 2, 2a, 3, 3a, 5, and 5a against complement activity increased in inverse proportion to number of free hydroxyls on B-ring of 5,7-dihydroxyflavone. Of the compounds tested, 4 showed the most potent inhibitory activity against the complement system. PMID- 16041766 TI - Free radical scavenging, enzyme inhibitory constituents from antidiabetic Ayurvedic medicinal plant Hydnocarpus wightiana Blume. AB - Hydnocarpus wightiana is advocated in traditional Indian medicine to possess strong antidiabetic activity. In the course of identifying bioactive fractions from Indian medicinal plants we observed that acetone extract of the seed hulls of H. wightiana possess strong free radicals (DPPH and ABTS) scavenging, alpha glucosidase and moderate N-acetyl-beta-D-glucosaminidase inhibitory activities. Further fractionation of the extract led to the isolation of hydnocarpin, luteolin and isohydnocarpin in substantial yields. All the compounds showed strong ABTS scavenging property. However, only luteolin could display strong DPPH scavenging activity. Furthermore, all the three compounds also showed varying degrees of alpha-glucosidase and N-acetyl-beta-D-glucosaminidase inhibitory activity, luteolin being the superior. The kinetics of alpha-glucosidase inhibition by these compounds showed that acetone extract inhibits the enzyme in competitive manner however, luteolin and isohydnocarpin showed mixed-type inhibition. This is the first report assigning hydnocarpin and isohydnocarpin free radical scavenging, alpha-glucosidase and N-acetyl-beta-D-glucosaminidase inhibitory properties and luteolin as N-acetyl-beta-D-glucosaminidase inhibitor. This study suggests that presence of amphiphilic antioxidant molecules along with enzyme inhibitory activities in the acetone extract of H. wightiana seed hulls may be responsible for the antidiabetic properties as advocated in traditional medicine. PMID- 16041767 TI - Central nervous system activity of yangambin from Ocotea duckei Vattimo (Lauraceae) in mice. AB - This work presents behavioral effects of yangambin isolated from the leaves of Ocotea duckei on open field, rota rod, barbiturate sleeping time, forced swimming and elevated plus maze test in mice. Yangambin was intraperitoneally administered to male mice at single doses of 12.5, 25 and 50 mg/kg. The results showed that yangambin in the doses of 25 and 50 mg/kg decreased the locomotor activity and the number of rearing. However, no change was observed in the rota rod test between the yangambin groups as compared to the control group. Reduction on the sleep latency and a prolongation of the sleeping time induced by pentobarbital was observed only with the yangambin dose of 50 mg/kg. In the forced swimming test, yangambin (25 and 50 mg/kg) increased the immobility time. Yangambin, in the doses of 25 and 50 mg/kg, decreased the number of entries and the time of permanence in the open arms of the elevated plus maze test. However, this effect can not be related to anxiogenic effects, but to a decrease in locomotor activity. The results showed that yangambin presents a depressant activity in the open field, forced swimming and pentobarbital sleeping time tests. These effects probably were not due to peripheral neuromuscular blockade, since there was no alteration on the rota rod test. Also, no anxiolytic effect was observed after the treatment with yangambin. PMID- 16041768 TI - Anti-HIV-1 activities in extracts from some medicinal plants as assessed in an in vitro biochemical HIV-1 reverse transcriptase assay. AB - An in vitro HIV-1 reverse transcriptase (RT) assay was used for screening of anti HIV activity of extracts obtained from some Kenyan medicinal plants. The assay utilises [3H]-methyl thymidine triphosphate (dTTP) as the enzyme substrate and polyadenylic acid.oligodeoxythymidylic acid [poly(rA).p(dT)(12-18)] as the template-primer dimmer. This assay was optimised and standardised with respect to the various experimental parameters in a microtiter plate methodology. The assay was then applied to test for potential antiviral activities of several Kenyan medicinal plant extracts and the concentrations producing 50% inhibition (IC50) of the HIV-1 RT were determined. This assay is described in this report and results obtained with some of the extracts are presented. PMID- 16041769 TI - Bioassay-guided isolation of antimicrobial benzopyrans and phloroglucinol derivatives from Hypericum species. AB - Recently the crude methanol extracts of six Hypericum species were analysed against a panel of microorganisms and it was found that H. caprifoliatum Cham. & Schlecht., H. myrianthum Cham. & Schlecht. and H. polyanthemum Klotzsch ex Reichardt were the most active. This paper reports the activity of the hexane, chloroform and methanol fractions of these plants as well as the activity of some isolated compounds against Staphylococcus aureus determined by bioautographic procedures. PMID- 16041770 TI - Dietary Garcinia cambogia does not modify skin properties of mice with or without excessive sucrose intake. AB - The influence of 3.3% Garcinia cambogia extract on the properties of mouse skin with or without 10% sucrose water loading was investigated. Mice (7-week-old) were given free access to a control diet or a diet containing Garcinia cambogia extract. They were also given water alone or both water and sucrose water. Their skin was compared by both biochemical and histological methods. The collagen and triacylglycerol contents were not significantly different among the four groups. Similarly, electron microscopy revealed no differences in the thickness of the dermis layer or the subcutaneous tissue layer. Mice given the diet containing Garcinia cambogia tended to have a reduced total number of adipocytes, but not significantly. These results suggest that Garcinia cambogia supplementation for at least 4 weeks does not induce a negative effect on skin properties in mice irrespective of excessive sucrose intake. PMID- 16041771 TI - Hystological and pharmacological study of Thymus piperella (L.). AB - Histological and pharmacological assays have been carried out with methanol, hexane, dichloromethane and butanol extracts of Thymus piperella (L.) leaves. All the extracts were considered innocuo in the toxicity test. Methanol and also hexane, dichlorometane and butanol fractions, inhibited significantly the contractions induced by acetylcholine in isolated rat ileum in a concentration dependent manner and the hexane extract was the most potent. However, the methanol extract did not modify the contractile effect of noradrenaline and histamine on isolated rat aorta and guinea-pig trachea respectively at the assayed dose (10, 100, 200 microg/mL). These results contribute to explain in part the use of this plant in folk medicine. In addition, morphological and histological structures characteristic of this species have been described for the first time. PMID- 16041772 TI - Structure-NMR chemical shift relationships for novel functionalized derivatives of quinoxalines. AB - (1)H, (13)C and (15)N NMR chemical shifts for a variety of novel quinoxalines were determined by different 2D methods and were calculated using the GIAO DFT approach. Comparison with experimental data shows good correlations in the case of (1)H, (13)C and (15)N chemical shifts. Different combinations of basis sets were tested. In non-polar solvents quinoxalines exist as dimers owing to strong hydrogen bonding. Calculations for dimers improve the correlation between experiment and theory. Additive empirical methods for estimating chemical shifts have drawbacks and have to be used with a great care for this type of compound. PMID- 16041773 TI - Characterization of polyphenols from plant materials through their silylation and 29Si NMR spectroscopy-line assignment through 29Si, 13C spin-spin couplings. AB - The lines in (29)Si NMR spectra of silylated polyphenols and some other compounds are difficult to assign owing to the absence of couplings with protons outside the silyl group. The assignment can be derived through small (n)J((29)Si, (13)C) couplings (n > 1). Using a previously described method for measurements of these couplings, the assignment procedure is demonstrated here on three examples of trimethylsilylated phenols: 7-hydroxyflavone, ferulic acid, and quercetin. In some cases the procedure can be used to identify carbon atoms to which the siloxy groups are attached. PMID- 16041774 TI - 6/7Li NMR study of the Li1-zNi1+zO2 phases. AB - A series of Li1-zNi1+zO2 materials have been synthesised by the coprecipitation route. An X-ray diffraction study was carried out on these materials using the Rietveld method to determine the departure from the ideal stoichiometry z, which ranges from 0 to 0.138. The actual Li/Ni ratio was also checked by chemical analyses using inductively coupled plasma (ICP) for each sample. The stoichiometric sample (z approximately 0) was obtained using a 15% Li excess. (6/7)Li NMR results from LiNiO2 (z approximately 0) show that the asymmetric shape of the NMR signal is due to anisotropy. Calculations give evidence that the paramagnetic dipolar interaction from the electron spins carried by Ni is anisotropic but does not completely explain the experimental anisotropy. (6)Li MAS NMR (magic angle spinning NMR) experiments and temperature standardisation NMR measurements unambiguously assign the isotropic position at +726 ppm. The static-echo NMR spectra of the non-stoichiometric Li1-zNi1+zO2 phases also exhibit an asymmetric shape whose width increases with the departure from the ideal stoichiometry z. (6/7)Li static and MAS NMR show that the 2zNi(2+) ions thus formed modify the dipolar interaction within the materials and also affect the Fermi contact interaction, since a distribution of Li environments is observed using (6)Li NMR for non-stoichiometric samples. PMID- 16041775 TI - Complete 1H and 13C NMR spectral assignment of symmetric and asymmetric bis spiropyran derivatives. AB - (1)H and (13)C NMR spectra of symmetric and asymmetric bis-spiropyrans, Series 1 3, were completely assigned. Especially, the (1)H assignment of asymmetric spiropyrans was achieved by utilizing (1)H-(1)H COSY and nOe experiments. All of the carbons in the dye molecules were investigated through a combination of heteronuclear 2D-shift correlation spectroscopy (HETCOR), together with an attached proton test (APT). PMID- 16041776 TI - Complete assignments of 1H and 13C NMR data for ten phenylpiperazine derivatives. AB - Ten phenylpiperazine derivatives were designed and synthesized. The first complete assignments of (1)H and (13)C NMR chemical shifts for these phenylpiperazine derivatives were achieved by means of 1D and 2D NMR techniques, including (1)H-(1)H COSY, HSQC and HMBC spectra. PMID- 16041777 TI - A great pioneer of clinical science remembered: introduction to the special issue in honor of Paul E. Meehl. AB - In this special issue, the seminal contributions to clinical psychology of Paul E. Meehl, who passed away in 2003, are commemorated. The nine articles comprising this special issue chronicle Meehl's remarkable intellectual biography and examine his influence on diverse domains of psychology, including the clinical versus actuarial prediction debate, the cognitive activity of the clinician, personality assessment and trait theory, the etiology of schizophrenia, the shortcomings of statistical significance testing, and the use of metascientific methods to evaluate competing models of human nature. These articles illustrate not only Meehl's legendary brilliance but also his pivotal role in forcing clinical psychologists to think more clearly and incisively about their subject matter. PMID- 16041778 TI - Paul Everett Meehl: the cumulative record. AB - In this brief biographical sketch of Paul Meehl, the authors examine the "cumulative record" of his scholarship and mentorship. This record sheds light on why Meehl is widely regarded as one of the most influential clinical psychologists of the 20th century, as well as on Meehl's remarkable intellectual life. Time has proven that Meehl's writings are exceptional in their quality, influence, breadth, and depth. In addition, Meehl's cumulative record raises important questions regarding the reinforcement contingencies in major research oriented psychology departments. PMID- 16041779 TI - Remembering and honoring Paul Meehl. AB - The author commemorates Paul Meehl by briefly presenting some of his cardinal contributions to clinical psychology, including the Minnesota Multiphasic Personality Inventory (MMPI), rational-emotive behavior therapy, and clinical versus statistical prediction. He also describes a few of his personal contacts with him. Meehl modeled a way of thinking, and thinking about thinking, that should be useful to all clinical psychologists. PMID- 16041780 TI - Clinical versus statistical prediction: the contribution of Paul E. Meehl. AB - The background of Paul E. Meehl's work on clinical versus statistical prediction is reviewed, with detailed analyses of his arguments. Meehl's four main contributions were the following: (a) he put the question, of whether clinical or statistical combinations of psychological data yielded better predictions, at center stage in applied psychology; (b) he convincingly argued, against an array of objections, that clinical versus statistical prediction was a real (not concocted) problem needing thorough study; (c) he meticulously and even-handedly dissected the logic of clinical inference from theoretical and probabilistic standpoints; and (c) he reviewed the studies available in 1954 and thereafter, which tested the validity of clinical versus statistical predictions. His early conclusion that the literature strongly favors statistical prediction has stood up extremely well, and his conceptual analyses of the prediction problem (especially his defense of applying aggregate-based probability statements to individual cases) have not been significantly improved since 1954. PMID- 16041781 TI - The ethical implications of Paul Meehl's work on comparing clinical versus actuarial prediction methods. AB - Paul E. Meehl's work comparing statistical versus actuarial prediction-and the large body of research that followed by others on the same topic-was mainly theoretical and empirical. Meehl also suggested that this work led to a "practical" conclusion, which was quite strong. The author argues that, in addition, it leads to an ethical conclusion, equally strong. Whether the scientific findings are combined with an overarching ethical principle that the best predictions possible should be made for clients, or whether these findings are framed as delineating what can be done for clients-and that clinicians ought not to attempt to do what they cannot-the conclusion is the same. Whenever statistical prediction rules (SPR's) are available for making a relevant prediction, they should be used in preference to intuition. Any modification of these rules should be systematic and subject to the same type of evaluation originally used to assess the SPR's themselves. It is even possible to develop near-optimal rules in new situations. Providing service that assumes that clinicians "can do better" simply based on self-confidence or plausibility in the absence of evidence that they can actually do so is simply unethical. PMID- 16041782 TI - In praise of clinical judgment: Meehl's forgotten legacy. AB - Although Paul E. Meehl demonstrated the limits of informal aggregation of data and prognostication by presumed experts, he remained convinced that clinical experience confers expertise of some kind. The authors explore this forgotten side of Meehl's legacy by reconsidering the validity of clinical judgment in its natural context, everyday clinical work. Three domains central to clinical practice are examined: diagnosis, interpretation of meaning, and intervention. It is argued that a more sanguine picture of clinical expertise emerges when the focus shifts from prediction at high levels of inference to (a) judgments at a moderate level of inference, (b) contexts for which clinical training and experience are likely to confer expertise, and (c) conditions that optimize the expression of that expertise (e.g., use of instruments designed for expert observers). The authors conclude by examining domains in which clinical judgment could prove useful in knowledge generation (e.g., hypothesis generation, identification of falsifying instances, item development). PMID- 16041783 TI - Essential Paul Meehl lessons for personality assessment psychology. AB - The author presents four essential Meehl lessons for personality assessment. First, Meehl's particular form of the integration of science and practice is described. Second, by outlining Meehl's Hedonic Capacity conjecture, Meehl's contribution to the inclusion of personality individual differences in generating the full clinical picture and in planning treatment is recognized. The third Meehl lesson is on the nature and importance of theory in test development and application programs. The fourth Meehl lesson is a more general epistemological lesson for psychology. Meehl's role in destroying the fantasy of an easy methodological formula for a scientific psychology is described. His program of taxometric research is shown to be an example of demanding greater material implications from theory. Meehl's corroboration index is described and contrasted with the p value of statistical significance testing. PMID- 16041784 TI - Paul E. Meehl's influence on experimental psychopathology: fruits of the nexus of schizotypy and schizophrenia, neurology, and methodology. AB - Paul E. Meehl made numerous contributions to clinical science and a hallmark of many of these contributions was their integrative nature. Meehl's positions on complex topics, especially one such as schizophrenia, were reflective of input from a variety of disciplines and levels of analysis. In this essay the authors focus on Meehl's uniquely rich contribution to our understanding of schizophrenia through his theoretical model of schizotypy, his abiding interest in exploring neurologically based indicators of schizophrenia liability and encouragement to others to pursue such indicators, and his passion for rigorous research methodology. Meehl's contributions in each of these areas continue to influence the direction and research strategies used in experimental psychopathology to illuminate the fundamental nature of schizophrenia. These contributions have also shaped inquiry into many other psychopathological entities. PMID- 16041785 TI - Meehl on metatheory. AB - This article proffers a critical survey of Paul E. Meehl's thoughts on the philosophy of science, as well as constructive criticisms of his views on corroboration of scientific theories. Using examples from clinical psychology and allied domains, six major topics are addressed: (a) the nature of theoretical constructs, (b) statistical diagnosis of clinical categories, (c) detection of hypothesized taxa, (d) null-hypothesis significance testing, (e) complexified hypothesis testing, and (f) cliometric theory appraisal. Through numerous examples, it is shown that Paul Meehl had a superb ability to recognize, articulate, and clarify hidden complexities, and other underappreciated obscurities in important working concepts. PMID- 16041786 TI - Why Paul Meehl will revolutionize the philosophy of science and why it should matter to psychologists. AB - Paul Meehl was a person of remarkable genius who made many seminal contributions to psychology and other fields. One of his most important, but less widely known potential contributions is the codevelopment, and the extension and elaboration of meta-science, or the science of science. Meta-science involves applying more rigorous methods (than are usually used) to the study of episodes in the history of science, or the historical track record, in order to address long-standing questions in the philosophy and history of science, aid in the selection of optimally effective methodology, and assist the scientist in higher level and complex integrative judgments (e.g., theory evaluation). Psychologists, given their methodological sophistication, can be major contributors to meta-scientific efforts and to the development of this field. PMID- 16041787 TI - Some remarks on characterization and application of stationary phases for RP-HPLC determination of biologically important compounds. AB - Biologically active compounds such as vitamins, steroids, nucleosides, peptides and proteins play a very important role in coordinating living organism functions. Determination of those substances is indispensable in pathogenesis. Their complex structure and physico-chemical properties cause many analytical problems. Chromatography is the most common technique used in pharmaceutical and biomedical analysis. The interaction between analyte and stationary phase plays a major role in the separation process. The structure of the packing has a significant influence on the results of the separation process. Various types of spectroscopic techniques, such as nuclear magnetic resonance spectroscopy, infrared spectroscopy, fluorescence spectroscopy and photoacoustic spectroscopy can be useful tools for the characterization of packings. Surface area measurements, elemental analysis, thermal analysis and microcalorimetric measurements are also helpful in this field. Part of the paper contains a description of chromatographic tests used for the determination of column properties. The description of the possibilities of surface characterization is not complete, but is focused on the most popular techniques and practical chromatographic tests. All the presented methods made possible the design and quality control of a new generation stationary phases, which are the future of high-performance liquid chromatography. PMID- 16041788 TI - A simple and sensitive assay for the quantitative analysis of paclitaxel and metabolites in human plasma using liquid chromatography/tandem mass spectrometry. AB - A sensitive and specific LC-MS/MS assay for the determination of paclitaxel and its 3'p- and 6-alpha-hydroxy metabolites is presented. A 200 microL plasma aliquot was spiked with a 13C6-labeled paclitaxel internal standard and extracted with 1.0 mL tert-butylmethylether. Dried extracts were reconstituted in 0.1 M ammonium acetate-acetonitrile (1:1, v/v) and 25 microL volumes were injected onto the HPLC system. Separation was performed on a 150 x 2.1 mm C18 column using an alkaline eluent (10 mm ammonium hydroxide-methanol, 30:70, v/v). Detection was performed by positive ion electrospray followed by tandem mass spectrometry. The assay quantifies a range for paclitaxel from 0.25 to 1000 ng/mL and metabolites from 0.25 to 100 ng/mL using 200 microL human plasma samples. Validation results demonstrate that paclitaxel and metabolite concentrations can be accurately and precisely quantified in human plasma. This assay is now used to support clinical pharmacologic studies with paclitaxel. PMID- 16041789 TI - Influence of implant/abutment joint designs on abutment screw loosening in a dental implant system. AB - The objective of this study was to investigate the influence of implant/abutment joint designs on abutment screw loosening in a dental implant system, using nonlinear dynamic analysis of the finite element method (FEM). This finite element simulation study used two dental implant systems: the Ankylos implant system (Degusa Dental, Hanau, German) with a taper joint (taper joint-type model), and the Branemark implant system (Nobel Biocare, Gothenburg, Sweden) with an external hex joint (external hex joint-type model). The nonlinear dynamic analysis was performed using three-dimensional finite element analysis. In comparing the movement of the taper type-joint model and external hex type-joint model, it was found that the external hex type-joint model had greater movement than the taper type-joint model. The external hex joint-type model showed rotation movement, whereas the movement of the taper joint-type model showed no rotation. It was concluded that the nonlinear dynamic analysis used in this study clearly demonstrated the differences in rotation of components in dental implant systems with taper or external hex joints. PMID- 16041790 TI - Effect of glass dissolution products on the detection of proteins by silver staining. AB - The influence of glass dissolution on the silver staining of proteins was investigated by reacting glass microspheres of varying chemical durability in boiling Laemmli sample buffer (LSB) for up to 5 min. All three of the investigated glass compositions leached Na+ ions to varying degrees during boiling in LSB, thereby causing an increase in the pH of the sample buffer. The LSB supernatant from the dissolution tests was mixed with unreacted LSB containing human serum albumin (HSA) and standard one-dimensional SDS-PAGE was performed. Silver staining was then used to visualize protein bands within the gel. The 30 Na2O.70 SiO2 glass exhibited pronounced degradation as shown by scanning electron microscopy. Further experiments employing solutions of neat LSB and reacted LSB (i.e., LSB containing glass dissolution products) mixed at varying ratios demonstrated the apparent significance of sample pH in affecting the inhibition of silver staining. The cause of this behavior may be due to an interference with the fixation stage of the staining protocol, thereby resulting in the loss of protein in subsequent rinsing stages. PMID- 16041791 TI - Evaluation of polymerization of light-curing hybrid composite resins. AB - The quality of polymerization of hybrid composite resins was tested to explore their feasibility for dental restorations. For this, microhardness, polymerization shrinkage, the coefficient of thermal expansion, and surface morphology were evaluated during or after light curing in conjunction with the thermocycling process. Each product had different microhardness values. The repeated thermal stimulus has no specific effect on the change of microhardness. The difference of microhardness between the thermocycled specimens and specimens stored only in distilled water was minor. The measured microhardness had a linear correlation with the filler content (vol %) of the tested specimens. The polymerization shrinkage had rapidly increased only during the light curing, and then it reached a plateau. Among the specimens, Z250 showed the least amount of shrinkage for all tested thicknesses. Regardless of the product, the shrinkage values increased as the specimens became thick. The coefficient of thermal expansion of the control specimens ranged between 42 and 55 microm/degrees C in the temperature range of 30-80 degrees C. The coefficient showed an inverse correlation with the filler content. Through the thermocycling process, Palfique Estelite showed randomly propagating cracks on the surface. Larger fillers showed a more apparent detachment than the smaller fillers. PMID- 16041792 TI - The medium-term results of treatment with hydroxyapatite implants. AB - Although the short-term results of implants with synthetic hydroxyapatite (HA), a bioactive material, have been favorable, few reports have been published concerning medium-term outcomes for this therapy. The authors recently analyzed data supplied by 37 medical facilities nationwide concerning the outcomes of synthetic HA implants [Bonfil (BF); Mitsubishi Materials Corporation, Tokyo] in 138 patients followed up for at least 5 years after treatment. When the data were analyzed, the patients were divided into two groups: the disease site-filling group (62 cases where bone defects created by disease were filled with synthetic HA) and the donor site-filling group (76 cases where bone defects created by bone donation were filled with synthetic HA). In the disease site-filling group, synthetic HA was used in combination with autologous bone or autogenous bone marrow donated from the same patient in 77% of cases. In the donor site-filling group, only synthetic HA was used in most cases. The average follow-up period after implantation was 7.9 years in the disease site-filling group and 9.1 years in the donor site-filling group. Therapy was rated radiographically as "very effective" or "effective" in 81% of patients in the disease site-filling group and in 89% of patients in the donor site-filling group. The therapy was not rated as "ineffective" in any case from either group. These results suggest that synthetic HA may serve as a very useful substitute for cancellous bone if used carefully, with its initial strength taken into account. PMID- 16041793 TI - Biomechanical properties of different segments of human umbilical cord vein and its value for clinical application. AB - No satisfactory effects have been obtained with the use of synthetic blood vessels (diameter <6 mm) as substitutes for human small arteries or veins for the purpose of clinical vascular reconstruction. Therefore, blood vessels of human origin, for example, umbilical cord blood vessels, with their wide availability, still should be considered. However, little information on biomechanical properties of human umbilical cord blood vessels is available. The objective was to provide a theoretical basis for the clinical application of umbilical cord veins as optional material for small-caliber grafts. This was a nonrandomized, noncontrolled in vitro study. The experiment was conducted in the Laboratory of Medical Biomechanics, Yunyang Medical College. Umbilical cord veins of 20 normal fetuses of spontaneous labor were collected by the Department of Obstetrics and Gynecology, Taihe Hospital in Shiyan City, Hubei Province. The fetuses aged 37-40 weeks, and the parturients were 20-30 years old. Umbilical cord veins of the 20 fetuses were used and the placental ends were treated as proximal ends while the fetal ends as distal ends. The fetal ends were divided into three segments: proximal, middle, and distal segments. The relationship between pressure of umbilical cord veins segments and the diameters was measured on the biomechanical experiment stand for soft tissues, and then the elastic modulus was calculated. The materials were transversely extracted, refrigerated, and sliced up before HE staining. The geometrical morphology indexes were measured by a computer image analysis system (Leica-Q500IW). The main outcome measures were: incremental elastic modulus (E(inc)), pressure-strain elastic modulus (E(p)), volume elastic modulus (E(v)), diameter, and wall thickness of the veins. E(inc), E(p), and E(v) of umbilical cord veins of proximal, middle, and distal segments increased with the pressure elevated. The three kinds of elastic modulus of proximal segments (E(inc): 26.98 +/- 3.21, E(p): 16.58 +/- 2.12, E(v): 8.31 +/- 2.35) were all lower than those of distal segments (E(inc): 33.20 +/- 4.21, E(p): 119.45 +/- 2.87, E(v): 9.71 +/- 1.32) (F = 95.74-126.52, p < 0.05), and a tendency to increase was shown from proximal segments to distal segments. Media thickness [(0.30 +/- 0.05)] mm, (0.24 +/- 0.03) mm] and the diameters [(3.07 +/- 0.12) mm, (2.30 +/- 0.13) mm] decreased gradually from proximal to distal segments (F = 12.76, p < 0.01). It is feasible to use umbilical cord veins as substitutes for the transplantation of small-caliber arteries in terms of basic biomechanical properties. On vascular grafting, different segments of umbilical cord veins should be chosen cautiously so that the biomechanical characteristics of umbilical cord vein grafts could be in accordance with those of host to increase the long-term patency rate of transplanted blood vessels. PMID- 16041794 TI - Endothelial cell reaction on a biological material. AB - The successful clinical application of materials should involve detailed investigations on interaction between them and tissue with which they will contact. We examined herein the behavior of endothelial cells (ECs) on a collagen material, using histological and immunohistochemical methods. We used isolated human umbilical cord vein cells (HUVECs) identified by means of endothelial specific antibodies. Cells were seeded in a standard density on a collagen membrane (Lycoll, Resorba, Nuernberg, Germany) and on gelatin-coated, control plastic surfaces, after two passages. These were then maintained for periods of 1, 7, or 14 days. The cells adhered, spread, and proliferated, and within 24 h started forming a subconfluent monolayer. We observed that the cultured cells expressed integrins (alpha5beta1 and alpha(v)beta3) and synthesized fibronectin. After 14 days, we could observe a confluent layer of ECs. We could conclude that the collagen material supported growth and attachment of endothelial cells. In addition, the attachment seemed to be most related to the fibronectin synthesized by the cells and to its highly expressed receptor (the alpha5beta1 integrin); even though this is not the only protein related to this adhesion, we observed that our cultured HUVECs did not synthesize vitronectin. PMID- 16041795 TI - Biocompatibility and degradation of poly(DL-lactic-co-glycolic acid)/calcium phosphate cement composites. AB - Injectable calcium phosphate (Ca-P) cement materials exhibit favorable osteocompatible behavior but are resorbed slowly because of a lack of a bone ingrowth-enabling macroporosity. In this study, poly(DL-lactic-co-glycolic acid) (PLGA) microparticles (average size 66 +/- 25 microm) were incorporated into Ca-P cement to obtain a macroporous Ca-P cement scaffold after PLGA hydrolysis in vivo. Preset PLGA/Ca-P cement composite discs of various weight ratios (0/100, 15/85, 30/70, and 50/50) were implanted subcutaneously and in cranial defects in rats for 12 weeks. Histological analysis revealed that all macropores in the PLGA containing composites (average pore size 73 +/- 27 microm) were filled with fibrous tissue and blood vessels (subcutaneous implants) and/or bone (cranial implants). Histologically, bone formation appeared most abundant and most consistent in the 30/70 PLGA/Ca-P cement composites. Histomorphometrical evaluation revealed a significant increase in defect fill in the 15/85 and 30/70 PLGA/Ca-P cement composites. Finally, subcutaneous and cranial 50/50 PLGA/Ca-P cement composites had degraded to a large extent, without adequate replacement by bone in the cranial implants. Therefore, we conclude that PLGA/Ca-P cement composites enable tissue ingrowth and show excellent osteocompatibility in weight ratios of 15/85 and 30/70 PLGA/Ca-P cement. In this model, 30/70 PLGA/Ca-P cement composites showed the most favorable biological response. PMID- 16041796 TI - Characterization of polyelectrolyte complexes between chondroitin sulfate and chitosan in the solid state. AB - Chondroitin sulfate (ChS) was used to form polyelectrolyte complexes with chitosan (ChI), and its potential as a colon-targeted drug carrier was investigated. In order to determine the optimal conditions for the formation of a stable polyelectrolyte complex, the formation of ChS/ChI complexes was examined at two different pH values with various weight ratios, or at a fixed molar ratio of ChS/ChI of 1/2 under various pH conditions. The molar compositions of the various ChS/ChI complexes were quantitated with the use of solid-state 13C CP MAS NMR. The equivalent molar ratios of the complexes ranged from 0.47 to 0.54, in agreement with the data determined by elemental analysis. The fact that these values were close to 0.5 suggests that most of the --OSO3- and the --COO- groups on ChS formed strong electrostatic interactions with the --NH3+ groups on ChI, obeying a simple stoichiometric reaction between two oppositely charged moieties. Similar compositions of the complexes were obtained under most conditions tested; however, different strengths of the interactions between the two polysaccharides were noted from measurements of the water-associated transition and thermal degradation temperatures and the degree of ChS dissolution. FTIR and 13C NMR clearly showed H-bond formation at low pH, indicating that in addition to the varying degrees of electrostatic interaction, H bonding may be involved in complex formation. The highest degradation temperature, as determined by thermal gravimetric analysis, and the lowest ChS sol fraction, as measured by gel permeation chromatography, were observed with the complex prepared at pH 5, with a 1:1 mole ratio of the two opposite charges in feed. This complex also exhibited the highest water-associated transition temperature, as determined by differential scanning calorimetry. Furthermore, the swelling behavior of these complexes was pH dependent; this is a property that can potentially be exploited to control drug release from these complexes under specific pH conditions. PMID- 16041797 TI - In vivo biostability of polyether polyurethanes with polyethylene oxide surface modifying end groups; resistance to biologic oxidation and stress cracking. AB - Polyethylene oxide (PEO) on polymer surfaces has been reported to reduce cellular adhesion, a very desirable property for cardiac pacing leads. A Shore 80A polyether polyurethane with up to 6% PEO surface-modifying end groups (SME) was evaluated for its chronic in vivo biostability. In a short-term (12 week) screening test, strained samples appeared to develop the same surface oxidation as unmodified polymer, but did not produce visible cracking > or =500x, prompting a longer-term study. By the time the longer-term study was initiated, most of the PEO SME had disappeared from the starting material's surface. After 1 year in vivo, surface oxidation, shallow surface cracking, and environmental stress cracking (ESC) developed on highly strained samples to the point of failure, so that there was no significant difference between the SME polymer and its control (the same polymer without SME). No further change was seen for up to 2 years of implantation. Unstrained PEO SME polymer developed shallow surface cracking, but no ESC up to 2 years of implantation. Thus, PEO SME slightly delayed, but did not stop biodegradation, and under unstrained conditions, has no adverse effect on biostability. PMID- 16041798 TI - Heterogeneous expression and regulation of hippocampal prostaglandin E2 receptors. AB - Although prostaglandin E2 (PGE2) has been shown to be critical to hippocampal synaptic signaling and neuronal survival, it is still not clear which subtypes of PGE2 receptors (EPs) are expressed and how these EPs are regulated in the hippocampus. To address these questions, the expression of the EPs was profiled in the hippocampus. Messenger RNAs and proteins of the four receptors, EP 1-4, were detected both in the hippocampus and in the neocortex. EP 2 and EP 3 appeared in greater abundance, whereas EP 1 and EP 4 were barely detectable. EP 1, EP 2 and EP 4 were mainly colocalized with synaptophysin, suggesting the presence of EP 1, EP 2, and EP 4 in presynaptic terminals. It appeared that interleukin-1 beta increased the expression of EP 2 and EP 4 mRNAs. A blockade of synaptic transmission with either tetrodotoxin or MK-801 plus 6,7 dinitroquinoxaline-2,3-dione (DNQX) for 6 hr increased EP 3 and EP 4 mRNA, whereas high K(+) (90 mM) or 4-aminopyridine enhanced EP 2 and EP 4. The EP 1 level did not change significantly under these conditions. The expressions of EP 2, EP 4, and EP 3 were further elevated or reduced in neurons treated with high K(+) for 24 hr. However, mRNA of EP 3 was down-regulated in neurons treated with tetrodotoxin or MK-801 plus DNQX for 24 hr. In addition, both EP 2 and EP 4 mRNAs were up-regulated within 4 hr after high-frequency stimulation associated with long-term potentiation induction in hippocampal slices. Our results indicate that the four EPs are heterogeneously expressed in the hippocampus, and their expression is differentially regulated by neuronal activities, suggesting that EPs may actively participate in hippocampal synaptic transmission and plasticity. PMID- 16041799 TI - Loss of dopaminergic neurons by the induction of inducible nitric oxide synthase and cyclooxygenase-2 via CD 40: relevance to Parkinson's disease. AB - A glial reaction associated with up-regulation of inflammatory molecules has been suggested to play an important role in dopaminergic neuron loss in Parkinson's disease (PD). Among inflammatory molecules, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) have been focused upon as key factors in the pathogenesis. However, the mechanism of how these molecules are induced in PD brains is not clearly understood. We focused on CD 40, which is expressed on neural cells and could be implicated in the neuroinflammation by inducing inflammatory molecules. We showed that both iNOS and COX-2 were up-regulated in microglia and astrocytes by CD 40 stimulation in association with a low dose of interferon-gamma (IFN-gamma) in vitro. Selective loss of dopaminergic neurons was induced by costimulation with CD 40 and IFN-gamma in mesencephalic cultures, which was protected by selective inhibitors of iNOS and/or COX-2. We also found in CD 40-stimulated astrocytes an increase of a low-affinity IgE receptor CD 23, which is known to induce iNOS expression. Together these data suggest that up regulated iNOS and COX-2 via the CD 40 pathway may lead to dopaminergic neuron loss and may participate in the neuroinflammaory pathway of PD. PMID- 16041800 TI - Evidence for macrophage-mediated myelin disruption in an animal model for Charcot Marie-Tooth neuropathy type 1A. AB - Charcot-Marie-Tooth neuropathy type 1A (CMT 1 A) is the most common inherited neuropathy in humans and is mostly caused by a 1.5-Mb tandem duplication of chromosome 17 comprising the gene for the peripheral myelin protein 22-kDa (PMP 22). Although there are numerous studies on the functional role of PMP 22, the mechanisms of myelin degeneration under PMP 22-overexpression conditions have not yet been fully understood. We have shown previously that in mouse mutants hetero- or homozygously deficient for two other myelin components, P0 and C x 32, respectively, immune cells contribute to the demyelinating neuropathy. To test this possibility for PMP 22 overexpression, we investigated a putative mouse model for CMT 1 A, i.e., the mouse strain C 6 1 mildly overexpressing human PMP 22 in peripheral nerves. Electron microscopic and electrophysiologic investigations revealed that this mouse strain develops pathologic features similar to those found in CMT 1 A patients. A novel finding, however, was the upregulation of CD8- and F4/80-positive lymphocytes and macrophages, respectively, in peripheral nerves. The observation that macrophages enter endoneurial tubes of the mutants and obviously phagocytose morphologically normal myelin strongly suggests that the myelin degeneration is mediated at least partially by these phagocytic cells. By gene array technology and quantitative RT PCR of peripheral nerve homogenates from PMP 22 mutants, monocyte chemoattractant protein-1 (MCP-1; cc l2) could be identified as a putative factor to attract or activate macrophages that attack myelin sheaths in this model of CMT 1 A. PMID- 16041801 TI - Variations in extracellular levels of dopamine, noradrenaline, glutamate, and aspartate across the sleep--wake cycle in the medial prefrontal cortex and nucleus accumbens of freely moving rats. AB - We used intracerebral microdialysis coupled with electrophysiologic recordings to determine relative changes in the concentrations of several neurotransmitters in the medial prefrontal cortex and nucleus accumbens of freely moving rats during waking, slow-wave sleep, and rapid eye movement (REM) sleep. The concentrations of noradrenaline, dopamine, glutamate, and aspartate in 2-min dialysate samples were analyzed by capillary electrophoresis combined with laser-induced fluorescence detection. Changes in glutamate and aspartate concentrations were found only in the nucleus accumbens, in which a decrease was obtained during both slow-wave sleep and REM sleep compared to waking. A progressive reduction in the release of noradrenaline was observed from waking to REM sleep in both structures. In contrast, dopamine concentrations were higher during waking and REM sleep compared to that during slow-wave sleep. The latter results demonstrate that contrary to the findings of earlier electrophysiologic studies carried out on ventral tegmental area dopaminergic neurons, changes in the release of dopamine in projection areas occur across the sleep-wake cycle. The elevated levels of dopamine during waking and REM sleep in the medial prefrontal cortex and the nucleus accumbens could result from changes during these two states in afferent modulation at the level of cell bodies or at the level of dopaminergic terminals. PMID- 16041803 TI - A systematic review of prevalence studies of dementia in Parkinson's disease. AB - Substantial variation in the prevalence of dementia in Parkinson's disease (PDD) has been reported. The aim of this study was to review systematically and critically previous studies of the prevalence of PDD using PubMed to search the literature. Studies focusing on PD and PDD, as well as those examining on the epidemiology of dementia subtypes, were included. Predefined inclusion and exclusion criteria were used and the quality of the studies included was rated. Articles were included if: (1) the proportion of PDD among patients with either PD or dementia was reported in an original study; (2) patients had been subjected to prospective clinical examination; and (3) strategies to include all subjects with either PD or dementia within the community or hospital clinics within a geographical area were employed. Twelve studies of the prevalence of PD or PDD (1,767 patients included) and 24 prevalence studies of dementia subtypes (4,711 patients included) met the inclusion criteria. In the PD/PDD studies, the proportion (mean and 95% confidence interval) with PDD in PD was 24.5% (17.4 31.5). There were significant methodological variations between studies and in the four studies that matched the quality criteria most closely, the rate of PDD was 31.1% (20.1-42.1). The prevalence of PDD was estimated as 0.5% in subjects 65 years or older. The percentage of PDD among those with dementia was 3.6% (3.1 4.1), with an estimated prevalence of PDD of 0.2% in subjects aged 65 years or older. Despite methodological variation, this systematic review suggests that 24 to 31% of PD patients have dementia, and that 3 to 4% of the dementia in the population would be due to PDD. The estimated prevalence of PDD in the general population aged 65 years and over is 0.2 to 0.5%. PMID- 16041802 TI - Changes in the amounts of chondroitin sulfate proteoglycans in rat brain after neonatal hypoxia-ischemia. AB - Chondroitin sulfate proteoglycans have been shown to participate in the pathogenesis of neuronal damages in the injured adult central nervous system (CNS). Upregulated expression of chondroitin sulfate proteoglycans has been reported around the injured sites and depletion of these chondroitin sulfate proteoglycans brings about increased axonal regeneration in the injured adult CNS. To examine if chondroitin sulfate proteoglycans are also involved in the pathologic process of hypoxia-ischemia in the neonatal brain, expressions of three chondroitin sulfate proteoglycans, neurocan, phosphacan, and neuroglycan C, were examined in rat brains after neonatal hypoxia-ischemia. Hypoxic-ischemic rats were produced by ligating the right carotid artery of 7-day-old rats, followed by 8% oxygen exposure. Western blot analysis revealed that in contrast to injured adult CNS, the amount of neurocan was reduced 24 hr after hypoxia in the neonatal hypoxic-ischemic cerebral hemisphere. The amounts of phosphacan and neuroglycan C were also reduced significantly 24 hr after hypoxia at the right injured cortex compared to those at the left cortex. Surprisingly, the immunohistologic staining for phosphacan was conversely intensified both at 24 hr and 8 days after hypoxia at the infarcted area. In addition, the habenula and fascicules retroflexus in the right cerebral hemisphere degenerated and became intensely immunostained with the anti-phosphacan antibody shortly after hypoxia. Hypoxic-ischemic insult may unmask phosphacan epitopes at the injured sites, resulting in intensified immunostaining. Because intensified immunostaining for neurocan and neuroglycan C was not observed, unmasking seems to be specific to phosphacan among these three chondroitin sulfate proteoglycans. PMID- 16041804 TI - FK 506 reduces tissue damage and prevents functional deficit after spinal cord injury in the rat. AB - We examined the efficacy of FK 506 in reducing tissue damage after spinal cord injury in comparison to methylprednisolone (MP) treatment. Rats were subjected to a photochemical injury (T8) and were given a bolus of MP (30 mg/kg), FK 506 (2 mg/kg), or saline. An additional group received an initial bolus of FK 506 (2 mg/kg) followed by daily injections (0.2 mg/kg intraperitoneally). Functional recovery was evaluated using open-field walking, inclined plane tests, motor evoked potentials (MEPs), and the H-reflex response during 14 days postoperation (dpo). Tissue sparing and glial fibrillary acidic protein (GFAP), biotinylated tomato lectin LEC, cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and interleukin 1 beta (IL-1 beta) immunoreactivity were quantified in the injured spinal cord. FK 506-treated animals demonstrated significantly better neurologic outcome, higher MEP amplitudes, and lower H-wave amplitude compared to that of saline-treated rats. In contrast, administration of MP did not result in significant differences with respect to the saline-treated group. Histologic examination revealed that tissue sparing was largest in FK 506-treated compared to saline and MP-treated animals. GFAP and COX-2 reactivity was decreased in animals treated with FK 506 compared to that in animals given MP or saline, whereas IL-1 beta expression was similarly reduced in both FK 506- and MP-treated groups. Microglia/macrophage response was reduced in FK 506 and MP-injected animals at 3 dpo, but only in MP-treated animals at 7 dpo with respect to saline injected rats. Repeated administrations of FK 506 improved functional and histologic results to a greater degree than did a single bolus of FK 506. The results indicate that FK 506 administration protects the damaged spinal cord and should be considered as potential therapy for treating spinal cord injuries. PMID- 16041805 TI - Decreased density of muscarinic receptors in the superior temporal gyrusin schizophrenia. AB - Recent studies have indicated that muscarinic receptors are involved in the pathophysiology in schizophrenia, particularly in cognitive deficits. The superior temporal gyrus (STG) is an area that has also been strongly implicated in the pathophysiology of schizophrenia. Therefore, in this study, we investigated the binding density of two muscarinic antagonists, [(3)H]pirenzepine and [(3)H]AF-DX 384, in the STG of schizophrenia patients compared with controls. A significant decrease (44% in the superficial layers and 48% in the deep layers, P<0.01) in binding density of [(3)H]pirenzepine was observed in schizophrenia patients, which suggested a reduction of muscarinic M1 and M4 receptor densities in the STG of schizophrenia patients. A tendency toward decreased [(3)H]AF-DX 384 binding density (34%, P=0.09) was also observed in schizophrenia patients compared with controls. Because of the positive correlation between [(3)H]pirenzepine and [(3)H]AF-DX 384 binding, and, insofar as both ligands have high affinities for the M4 receptor, the involvement of M4 receptor alteration is also suggested in the STG in schizophrenia. These results suggest that changes of the muscarinic receptors M1 and M4 might contribute to the STG pathology in schizophrenia. PMID- 16041806 TI - Continuous operation of foamed emulsion bioreactors treating toluene vapors. AB - Continuous operation of a new bioreactor for air pollution control called the foamed emulsion bioreactor (FEBR) has been investigated. The effect of several liquid feeding strategies was explored. The FEBR exhibited high and steady toluene removal performance (removal efficiency of 89%-94%, elimination capacity of 214-226 g/m3h at toluene inlet concentration of 1 g/m3) for up to 360 h, when 20% of the culture was replaced every 24 h by a nutrient solution containing 4 g/L of potassium nitrate as a nitrogen source. This feeding mode supported a high cell activity measured as INT reduction potential and active cell growth without being subject to nitrogen limitation. In comparison, operating the FEBR with the liquid in a closed loop (i.e., batch) resulted in a significant decrease of both the removal efficiency of toluene and INT reduction activity. Operation with feeding active cells resulted in stable and effective treatment, but would require a significant effort for mass culture preparation. Therefore, the continuous process with periodically feeding nutrients was found to be the most practical and effective operating mode. It also allows for stable operation, as was shown during removal of low concentration of toluene or after pollutant starvation. Throughout the study, INT reduction measurements provided insight into the process. INT reduction activity data proved that under normal operating conditions, the FEBR performance was limited by both the kinetics and by mass transfer. Overall, the results illustrate that engineered gas-phase bioreactors can potentially be more effective than conventional biofilters and biotrickling filters for the treatment of air pollutants such as toluene. PMID- 16041807 TI - Tangential flow microfiltration and ultrafiltration for human influenza A virus concentration and purification. AB - Large scale purification of viruses and viral vectors for gene therapy applications and viral vaccines is a major separation challenge. Here tangential flow microfiltration and ultrafiltration using flat sheet membranes has been investigated for concentration of human influenza A virus. Ultrafiltration membranes with molecular weight cutoffs of 100 and 300 kDa as well as 0.1, 0.2 and 0.45 microm microfiltration membranes have been tested. The results indicate that use of 300 kDa membranes not only concentrate the virus particles but also lead to a significant removal of host cell proteins and DNA in the permeate. Tangential flow filtration may be used to fractionate virus particles. Human influenza A virus particles are spherical with an average size of 100 nm. Use of a 0.1 microm membrane leads to passage of virus particles less than 100 nm into the permeate and an increase of larger particles in the retentate. These results suggest that control of the transmembrane pressure, membrane pore size and pore size distribution could enable isolation of intact virus particles from damaged virions. Isolation of the virus particles of interest from viral fragments and other particulate matter could result in simplification of subsequent purification steps. Larger pore size membranes such as 0.45 microm that allow the passage of all virus particles may be used to remove host cell fragments. In addition virus particles attached to these fragments will be removed. Careful selection of membrane morphology and operating conditions will be essential in order to maximize the benefit of tangential flow filtration steps in the purification of viral products from cell cultures. PMID- 16041808 TI - Homo-/heterotrinuclear mixed-valent oxo-centered iron/nickel clusters-Mossbauer studies on internal electron-exchange processes. AB - In a one-pot reaction of N-(5-methylthiazole-2-yl)-thiazole-2-carboxamide HL2 (3) with iron(II) acetate in air, the homotrinuclear heteroleptic mixed-valent oxo centered iron cluster [Fe(II)Fe(III)O(L2)3(OAc)3] (4) was formed. Exchange of iron(II) in 4 by nickel(II) afforded the heteronuclear cluster [Ni(II)Fe(III)O(L2)3(OAc)3] (6). To obtain crystals suitable for X-ray structure analyses, in 4 and 6, the OAc- co-ligands were exchanged by OBz- ligands to give [Fe(II)Fe2(III)O(L2)3(OBz)3] (5) and [Ni(II)Fe(III)O(L2)3(OBz)3] (7). The complexes 5 and 7 are isostructural and made up of three ditopic, tridentate ligands (L2)- and three bridging benzoate co-ligands, which fix the three metal ions in the corners of a triangle with an mu3-O2- ion in the center. The mixed valent character of 4-7, their intramolecular electron-exchange processes, and their redox properties were studied by variable-temperature Mossbauer spectroscopy and cyclic voltammetry. PMID- 16041809 TI - A small cavity with reactive internal shell atoms spanned by four {As(W/V)9}-type building blocks allows host-guest chemistry under confined conditions. AB - The reaction of [H2As(III)W18O60]7- with VO2+ and SO4(2-) ions in aqueous solution leads to a V(IV)/V(V) mixed-valence cluster anion containing the {As4M40O140}-type cryptand which has a high formation tendency. An important result is that it exhibits a new type of reactive internal cavity shell. The correspondingly obtained compound Na(NH4)20[{(V(IV)O(H2O))(V(IV)O)2(SO4)2}{(As(III)W9O33)2(As(III)W7.5V(V)1.5O31)2( O2)4}] x 40 H2O (1), which can also be synthesized from a precursor with the preorganized cryptand, was characterized by elemental and thermogravimetric analyses (determination of crystal water content), redox titrations (determination of the number of V(IV) centers), electronic absorption as well as vibrational spectra, single-crystal X-ray structure analysis (including bond valence sum calculations), and magnetic susceptibility measurements. The relatively small central cavity--formed by the linking of four {AsM9}-type lacunary units (M = W/V) by four WO6 octahedra--allows positioning of a variety of cationic as well as anionic "guests" under confined conditions according to a new approach: replacement of some of the W by V atoms leads to high reactivity of the internal cavity shell as a result of relatively weak VO bonds compared to the WO bonds. This allows an interesting "encapsulation chemistry" with new options. In the present case the cavity contains besides an arrangement of three V(IV) centers, two sulfate groups that replace O atoms of the {AsM9} units as well as an interesting hydrogen bond situation. PMID- 16041810 TI - Forster-type energy transfer along a specified axis. PMID- 16041811 TI - Hierarchically structured carbon: synthesis of carbon nanofibers nested inside or immobilized onto modified activated carbon. PMID- 16041812 TI - Paramagnetic platinum-rhodium octamers bridged by halogen ions to afford a quasi 1D system. PMID- 16041813 TI - A DNA-protein nanoengine for "on-demand" release and precise delivery of molecules. PMID- 16041814 TI - Supramolecular click chemistry with a bisammonium-C60 substrate and a ditopic crown ether host. PMID- 16041816 TI - Hexakis(trimethylsilyl)tetrahedranyltetrahedrane. PMID- 16041815 TI - Highly enantioselective Cu-catalyzed conjugate additions of dialkylzinc reagents to unsaturated furanones and pyranones: preparation of air-stable and catalytically active Cu-peptide complexes. PMID- 16041817 TI - Dithiaethyneporphyrin: an atypical [18]triphyrin(4.1.1) frame for contracted porphyrins. PMID- 16041818 TI - Low-spin state structure of [Fe(chloroethyltetrazole)6](BF4)2 obtained from synchrotron powder diffraction data. AB - The complex [Fe(teec)6](BF4)2 (teec = chloroethyltetrazole) shows a two-step complete spin-crossover transition in the temperature range 300-90 K. Time resolved synchrotron powder diffraction experiments have been carried out in this temperature range, and crystal structure models have been obtained from the powder patterns by using the parallel tempering technique. Of these models, the low-spin state structure at 90 K has been refined completely with Rietveld refinement. Its structural characteristics are discussed in relation to the high spin state model and other spin-crossover compounds. The complex shows a remarkable anisotropic unit-cell parameter contraction that is dependent on the applied cooling rate. In addition, the possible important implications for the interpretation of spin-crossover behavior in terms of structural changes are discussed. PMID- 16041819 TI - Synthesis, photophysics, and photoresponse of fullerene-based azoaromatic dyads. AB - The synthesis and photophysical characterization of a series of fullerene-based, donor-acceptor dyads is presented, along with a description of their behavior as single molecular components in photovoltaic cells. The spectroscopic and photophysical properties of the dyads, investigated by steady-state fluorescence spectroscopy, pico- and nanosecond transient optical spectroscopy and time resolved electron paramagnetic resonance (EPR) spectroscopy, revealed that the dyads undergo multiple-step energy transfer from the donor singlet excited state to the fullerene triplet excited state, which in turn decays to the donor triplet state. The inefficient formation of a charge-separated state, both in solution and in the solid state, translates into a poor photovoltaic performance of dyads 2 b-4 b if compared to that of dyad 1 b, in which photoinduced electron transfer is operative in the solid state. In addition, the results of the photophysical investigation suggested that the performance of the solar cells was also limited by the low-lying donor triplet excited state that acts as a photoexcitation energy sink. PMID- 16041820 TI - New fragmentation processes of pyridin-2(1H)-ones upon electron impact ionization. PMID- 16041821 TI - Characteristic pH and electrolyte concentration dependences of 2-aminopyridine derivatized oligosaccharides (N-glycans) in sonic-spray ionization mass spectrometry. AB - The intensities of ion signals from neutral oligosaccharides (N-glycans) derivatized with 2-aminopyridine (PA) were analyzed by ion trap mass spectrometry with a sonic-spray ionization (SSI) source, in both positive- and negative-ion modes, while varying the pH and concentration of ammonium acetate buffer solution. Two characteristic results are reported and discussed. The first characteristic is the pH dependence of the ion intensities; on increasing the solution pH from 4.3 to 8.6, positive ion intensities increase and negative ion intensities decrease. The second characteristic concerns the dependence of ion intensities on electrolyte concentration; on increasing the electrolyte concentration, the SSI efficiency for the PA N-glycans first increases and then decreases. Assuming that the SSI mechanism essentially conforms to the statistical charging model and the charge residue model, a new model that focuses a great deal of attention on the counter (electrolyte) ion distribution surrounding the solvated analyte (PA N-glycan) is proposed, in particular to rationalize the characteristic pH dependence. PMID- 16041822 TI - A rapid sample preparation method for mass spectrometric characterization of N linked glycans. AB - A rapid method for analysis of glycans of glycoproteins is presented. This method comprised deglycosylation, sample cleanup and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis of glycans. The enzymatic deglycosylation of N-linked glycoproteins was enhanced in terms of speed and reproducibility using an enzyme-friendly surfactant. The released glycans were desalted using a micro-scale solid phase extraction (SPE) device packed with a hydrophilic interaction chromatography (HILIC) sorbent. Hydrophilic glycans were well retained by SPE, while salts and surfactants were removed from the sample. The glycans were eluted using 25-50 microL of solvent and analyzed directly without derivatization using MALDI-MS. MALDI quadrupole time-of-flight (Q-Tof) instrumentation was utilized for glycan profiling and structure characterization by tandem mass spectrometry (MS/MS). The presented method allows sensitive analysis of glycans benefiting from optimized deglycosylation reactions and efficient sample cleanup. PMID- 16041823 TI - Mass spectrometric investigation of gaseous YbH, YbO and YbOH molecules. AB - The high-temperature gaseous molecules YbH, YbO and YbOH have been identified and their thermochemistry investigated by the Knudsen effusion mass spectrometry technique coupled with a controlled pressure gas inlet system. Solid ytterbium monosilicide and disilicide samples were made to react in the Knudsen cell with H2(g) and H2(g)/O2(g); in these conditions, several gaseous species (Yb, YbO, YbH, YbOH, SiO, SiO2, H2O) were formed under equilibrium conditions. The temperature dependences of the partial pressures of the observed gaseous molecules were analyzed to derive the Yb--X bond energies (X = H, O, OH). Selected values are D0o(Yb--H) = 179.4 +/- 2.0 kJ mol(-1), D0o(Yb--O) = 384 +/- 10 mol(-1) and D0o(Yb--OH) = 322 +/- 12 kJ mol(-1), and Delta(at)H0o(YbOH) = 746 +/- 12 kJ mol(-1). Density functional theory (DFT) calculations were also performed. Experimental and computational results are discussed and compared to previous data when available. The SiO/SiO2 high-temperature gaseous equilibrium was also observed. PMID- 16041824 TI - Analysis of vitamin E derivatives in serum using coordinated ion spray mass spectrometry. AB - A method for the extraction and analysis of tocopherols from serum using coordinated ion spray (CIS) mass spectrometry was developed and tested. The tocopherols were extracted from serum and analyzed by direct infusion into the mass spectrometer, bypassing the need for a liquid chromatography step. CIS is a method for improving the ionization efficiency of non-polar compounds by adding metal ions to the electrospray solvent. The non-polar analytes appear as metal adducts in the resulting mass spectrum. Silver was used as the metal ion for the CIS, causing analyte masses to be increased by 107 and 109 Da from the two main silver isotopes. Vitamin E succinate was added to the samples before extraction and was used as an internal standard to compensate for any variations in the extraction efficiency or mass spectrometric response. alpha-Tocopherol and an ether-linked analogue known as alpha-TEA were analyzed in concentrations from 1.25-40 microg/mL (1.9-60 pg consumed). The response curve was constructed by comparing the response of the analytes to the internal standard and gave linear results with r2 values greater than 0.98. This new method was shown to be sensitive, reproducible, fast and required very small amounts of analyte. PMID- 16041825 TI - Application of electrospray ionization with Fourier transform ion cyclotron resonance mass spectrometry for structural screening of core oligosaccharides from lipopolysaccharides of the bacteria Proteus. AB - Electrospray ionization with Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICRMS) was used for screening and structural elucidation of core oligosaccharides isolated from lipopolysaccharides of bacteria of the genus Proteus. Mass spectra allowed the determination of the molecular masses with high accuracy and the estimation of the chemical heterogeneity of the samples. They did not, however, provide sufficient information to identify structural details of the branched oligosaccharides. Therefore, various fragmentation techniques for determining such details were examined. Infrared multiphoton dissociation tandem mass spectrometry (IRMPD-MS/MS) experiments in negative ion mode resulted in cleavage between the structurally conserved inner core region and the variable outer core region. Positive ion capillary skimmer dissociation mass spectra showed numerous fragment ion peaks, including those corresponding to the subsequent cleavage of the glycosidic linkages starting from the non-reducing end of the oligosaccharide. Despite their complexity, these mass spectrometric studies allowed confirmation of previously determined Proteus lipopolysaccharide core structures, and identification of new related structures in other strains of these bacteria. PMID- 16041826 TI - High-precision oxygen and carbon isotope analysis of very small (10-30 microg) amounts of carbonates using continuous flow isotope ratio mass spectrometry. PMID- 16041827 TI - Study of the factors influencing oxazolone ring formation from alpha-benzamido methyl esters: differences between electrospray and atmospheric pressure chemical ionization. PMID- 16041829 TI - Stability of decisional role preference over the course of cancer therapy. AB - Cancer patients vary in their preferred level of involvement in medical decision making, and responding to patients' desired level of involvement is a key element of good medical care. While the literature has clearly demonstrated heterogeneity among cancer patients' preferences, less is known about how the preferences of any given patient may change over time. This longitudinal study compared cancer patients' preferences for involvement in medical decision making from the time of diagnosis to the time of completion of therapy. Data from 729 cancer patients with mixed diagnoses were analyzed. Most patients reported a change in preferred level of involvement over time, and multivariate analysis demonstrated that patients tend to prefer a decreasing level of involvement over time (p<0.0001). Stability of patients' preferences was also associated with type of cancer, but not with other sociodemographic characteristics. The results from this study highlight the importance of reevaluating patients' preferences for involvement in medical decision making throughout the course of cancer therapy, as such preferences are likely to change. PMID- 16041830 TI - Contribution of three-dimensional volume contrast imaging to the sonographic assessment of the fetal uterus. AB - OBJECTIVE: To investigate the contribution of volume contrast imaging (VCI) in assessing the fetal uterus in normally developed female fetuses. METHODS: The pelvis of 38 normal female fetuses was examined at 20-22 and 32-34 weeks' gestation using both conventional two-dimensional (2D) ultrasound and VCI on the same transverse or oblique longitudinal view of the fetal pelvis. Two experienced sonographers evaluated the ability of both techniques to image the fetal uterus. Results were compared by kappa index to evaluate the interobserver variability. RESULTS: A clear picture of the fetal uterus was obtained in 50% and 82-87% of the cases at 20-22 weeks' gestation and in 80-85% and 95-100% of the cases at 32 34 weeks' gestation using conventional 2D ultrasound and VCI, respectively. There was moderate to good agreement of uterus visualization between the two observers, with kappa values ranging from 0.43 to 0.65. The lower level of agreement was obtained for conventional 2D ultrasound during the second trimester. CONCLUSIONS: Our results suggest that VCI may be successfully applied to prenatal ultrasonography of the fetal pelvis anatomy. By enhancing the contrast between the intrapelvic organs, VCI provides a clearer picture of the fetal uterus. PMID- 16041831 TI - Factors influencing the success of conservative treatment of interstitial pregnancy. AB - OBJECTIVE: To identify demographic, morphological and biochemical characteristics of interstitial pregnancies that are associated with a successful outcome of conservative treatment (expectant management or medical treatment with methotrexate). METHODS: Over a period of 6 years all clinically stable women with a certain ultrasound diagnosis of interstitial ectopic pregnancy were managed either expectantly (no interventions) or medically (systemic or local methotrexate injection). The outcome of conservative treatment was compared to a number of diagnostic parameters, which were recorded at the initial visit. The treatment was classified as successful if serum human chorionic gonadotropin (hCG) level declined below 20 IU/L without the need for any additional interventions, such as the administration of methotrexate or surgery. RESULTS: A total of 42 interstitial pregnancies were diagnosed during the study period. Out of 35 women included in the final analysis 7 (20%) were managed expectantly and 28 (80%) received either local (n = 23) or systemic methotrexate (n = 5). There were no significant differences in the success rates of expectant treatment and treatment with either systemic or local methotrexate. The initial median serum hCG was significantly lower in women with successful conservative management (3216 IU/L vs. 15 900 IU/L; P < 0.05) but there were no other significant differences between cases with successful and failed treatment. CONCLUSION: The measurement of serum hCG at the initial visit may be used to predict the likelihood of successful conservative treatment of interstitial pregnancy. PMID- 16041832 TI - Use of lipoplex-induced nuclear factor-kappaB activation to enhance transgene expression by lipoplex in mouse lung. AB - BACKGROUND: Although lipofection-induced TNF-alpha can activate nuclear factor kappaB (NF-kappaB), which, in turn, increases the transgene expression from plasmid DNA in which any NF-kappaB responsive element is incorporated, no attempts have been made to use such biological responses as NF-kappaB activation against a vector to enhance vector-mediated gene transfer. METHODS: A lipoplex composed of N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium and cholesterol liposome and plasmid DNA encoding firefly luciferase under the control of the cytomegalovirus immediate early promoter (pCMV-Luc) was intravenously injected into mice. Luciferase activity as well as NF-kappaB activation in the lung were evaluated. Then, a novel plasmid DNA, pCMV-kappaB-Luc, was constructed by inserting 5 repeats of NF-kappaB-binding sequences into the pCMV-Luc. RESULTS: NF kappaB in the lung was activated by injection of the lipoplex and its nuclear localization was observed. An injection of lipopolysaccharide 30 min prior to the lipofection further activated NF-kappaB. At the same time, the treatment significantly increased the transgene expression by lipoplex, suggesting a positive correlation between expression and NF-kappaB activity. Based on these findings, we tried to enhance the lipoplex-based transgene expression by using NF kappaB activation. The lipoplex consisting of pCMV-kappaB-Luc showed a 4.7-fold increase in transgene expression in the lung compared with that with pCMV-Luc. CONCLUSIONS: We demonstrated that NF-kappaB activation by lipoplex can be used to enhance lipoplex-mediated transgene expression by inserting NF-kappaB-binding sequences into plasmid DNA. These findings offer a novel method for designing a vector for gene transfer in conjunction with biological responses to it. PMID- 16041833 TI - The polygenetically inherited metabolic syndrome of WOKW rats is associated with insulin resistance and altered gene expression in adipose tissue. AB - BACKGROUND: Wistar Ottawa Karlsburg W (RT1u) rats (WOKW) develop a complete metabolic syndrome closely resembling the human disease. The aim of this study was to characterize the phenotype of adipose tissue in WOKW rats with regard to adipocyte metabolism, insulin resistance, and gene expression and thus to define the phenotype more precisely. METHODS: Glucose metabolism, insulin sensitivity, and gene expression of key adipocyte genes, including adiponectin, interleukin 6 (Il6), 11 beta-hydroxysteroid dehydrogenase (11beta Hsd), peroxisome proliferator activated receptor gamma (Ppar gamma), forkhead box O1 (Foxo1), glucose transporter 4 (Glut4), CCAAT/enhancer binding protein (C/ebp alpha), and fatty acid synthase (Fasn) were characterized in adipocytes from epididymal and subcutaneous fat depots of 28-week-old male WOKW rats and Dark Agouti (DA) controls. RESULTS: WOKW rats display decreased insulin-stimulated glucose uptake and decreased insulin sensitivity during lipogenesis and lipolysis in isolated adipocytes. The severe insulin resistance predominantly in epididymal adipose tissue of WOKW rats is associated with a 10-fold decrease in adipocyte adiponectin gene expression, decreased Ppar gamma, but increased Foxo1 gene expression compared to DA rats. CONCLUSIONS: Insulin resistance in adipose tissue is associated with altered adipocyte gene expression in WOKW rats, additionally completing the picture of the metabolic syndrome in this animal model. This fact not only qualifies the WOKW rat for further detailed analysis of genetic determinants of metabolic syndrome but also highlights its suitability for pharmacological research. PMID- 16041834 TI - Design, synthesis, and preliminary evaluation of gabapentin-pregabalin mutual prodrugs in relieving neuropathic pain. AB - As a part of a program for the development of specific analgesics in relieving neuropathic pain, the purpose of the present study was to investigate a new concept that involves the conjugation of two drugs, gabapentin and pregabalin, as mutual prodrugs using a chemical modification approach. A series of gabapentin pregabalin diester compounds were synthesized using linear or branch bis-hydroxyl linkers. Their pharmacological properties for treating neuropathic pain were investigated in a rat model of chronic sciatic nerve constriction injury (CCI). In-vivo evaluation demonstrated that 1a and 1b composed of two gabapentin molecules as well as 3a composed of gabapentin and pregabalin with the short linear linker, were effective in reversing tactile allodynia in CCI rats. Compounds with longer or side-branched linkers showed lower efficiencies and severe adverse effects. PMID- 16041835 TI - Synthesis of some floctafenine derivatives of expected anti inflammatory/analgesic activity. AB - New derivatives related to floctafenine were synthesized and representative examples were screened for their anti-inflammatory and analgesic activities. All compounds tested were found to exhibit anti-inflammatory and analgesic activities and some were more potent than the references, floctafenine and indomethacin, in carragenan-induced rat's paw edema. None of the tested compounds showed an ulcerogenic effect on the p-benzoquinone-induced writhing test in mice. PMID- 16041836 TI - Convenient synthesis of fused heterocyclic 1,3,5-triazines from some N-acyl imidates and heterocyclic amines as anticancer and antioxidant agents. AB - N-Acyl imidates (2), reacting with 5-amino pyrazole (3), 2-aminobenzimidazole (4), 3-amino-1,2,4-triazole (5), 3,5-diamino-1,2,4-triazole (6), and 5 aminotetrazole (7) give pyrazolo[1,5-a][1,3,5]triazine (8), benzo[4,5]imidazo[1,2 a][1,3,5]triazine (9), [1,2,4]triazolo [2,3-a][1,3,5]triazine (10), [1,2,4]tri azolo[2,3-a][1,3,5]triazin-5-ylamine (12), and tetrazolo-[1,5-a][1,3,5]triazine (14) derivatives, respectively. The synthesized compounds were characterized on the basis of IR, (1)H-NMR, (13)C-NMR, and mass spectral data and elemental analyses results. Five of the newly synthesized compounds, 8a, 9a, 10a, 12a, and 14a, were selected by National Cancer Institute and screened for their anticancer activity against three cancer cell lines MCF7, NCI-H460, and SF-268, where 12a exhibited moderate anti-proliferation potential. 12a was, thus, further tested for anticancer activity against 60 human cancer cell lines and showed moderate growth inhibition potency. 12a showed a high growth inhibitory activity against A498 renal cancer cell line. All of the newly synthesized compounds 8-10, 12 and 14 were tested for their antioxidant capacity where they exhibited very high activity, even higher than the widely used reference antioxidants butylated hydroxytoluene and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox). Compound 12a also showed the highest antioxidant activity. PMID- 16041837 TI - A note on the antitubercular activities of 1-aryl-5-benzylsulfanyltetrazoles. AB - A set of 32 1-phenyl-5-benzylsulfanyltetrazoles substituted on the phenyl ring as well as on the benzyl moiety was synthesized. The compounds were evaluated for in vitro antimycobacterial activity against Mycobacterium tuberculosis. The activity against M. tuberculosis becomes higher with increasing electron-accepting properties of the substituents on the phenyl ring. On the other hand, any substitution on the benzylic moiety decreases the activity. PMID- 16041838 TI - Synthesis of some new 2-(6-methoxy-2-naphthyl)- 5-aryl-1,3,4-oxadiazoles as possible non-steroidal anti-inflammatory and analgesic agents. AB - The synthesis of some new 2-(6-methoxy-2-naphthyl)-5-aryl-1,3,4-oxadiazoles (4a k) has been described. Ethyl-6-methoxy-2-naphthoate (1) yielded on treatment with hydrazine hydrate to 6-methoxy-2-naphthoic acid hydrazide (2). Compound 2 reacted with substituted aromatic carboxylic acids (3a-k) in phosphorus oxychloride yielded 2-(6-methoxy-2-naphthyl)-5-aryl-1,3,4-oxadiazoles (4a-k). Newly synthesized compounds were characterized by IR, (1)H-NMR and mass spectral data. All the compounds were screened for their anti-inflammatory and analgesic activity. Compound 4j exhibited promising anti-inflammatory and analgesic activities. PMID- 16041839 TI - Chromatin modifications as targets for new anticancer drugs. AB - Chromatin proteins undergo diverse posttranslational modifications, esp. acetylation and methylation, that contribute to the control of transcriptional processes. The result of these modifications in its various states is called the histone code. This review presents an overview of those modifications of chromatin proteins that affect the side chains of lysines and arginines and define variations of the chromatin acetylome and methylome. The relevant enzymes are presented and the feasibility to influence their activity by inhibition or activation is discussed. The manipulation of these enzymes is an exciting strategy towards an increased understanding of their role in the functionality of a cell. Additionally, this may lead to new approaches for the treatment of diseases that are based on a dysregulation of transcription, especially cancer. PMID- 16041840 TI - An open-label study of the effects of bupropion SR on fatigue, depression and quality of life of mixed-site cancer patients and their partners. AB - This preliminary study investigated whether bupropion sustained release (SR) improved symptomatic fatigue, depression and quality of life in cancer patients and caregiver quality of life. The sample consisted of a prospective open case series of 21 cancer patients, with fatigue and with or without depression at moderate to severe levels, referred for psychiatric assessment from a tertiary care cancer centre. Both patient symptom ratings and caregiver ratings were measured before and after 4 weeks of treatment with the maximally tolerated dose of bupropion in the range of 100-300 mg per day. At trial completion, significant improvement was found for symptoms of fatigue and depression. Subjects were divided into two groups: depressed and non-depressed (based on a cut-off score of 17 on the Hamilton Depression Rating Scale). Both groups reported improvement for fatigue and depressive symptoms. Depressed subjects and their caregivers did not experience any change in quality of life, while the non-depressed subjects and their caregivers reported improvements. Results from this small group of patients suggest that bupropion may have potential as an effective pharmaceutical agent for treating cancer-related fatigue. A randomized, placebo-controlled trial with this medication is indicated. PMID- 16041841 TI - Cancer-related health worries and psychological distress among older adult, long term cancer survivors. AB - While long-term survivors (5 years+) do not face the stressors of diagnosis and treatment, they continue to face the uncertainties that survivorship brings: recurrence, other cancers, late effects of treatment, and the potential of a shortened life expectancy. This research focuses on the cancer-related health worries of older adult, long-term cancer survivors, the factors that predict these worries, and their link to traditional measures of psychological distress. Specifically, a model is proposed that identifies the personal (including race and gender) and illness/treatment characteristics of survivors that are significantly associated with cancer-related health worries and their effects on anxiety and depression. Descriptive and multivariate analyses of a random sample of 321 long-term survivors in a major cancer center tumor registry are used to address these issues. About one-third of survivors continue to report worries about recurrence, worries about a second cancer, and worries that symptoms they experience may be from cancer. The regression analyses show that cancer-related health worries is a significant predictor of both depression (beta=0.36) and anxiety (beta=0.21). Race is a significant predictor; being African American is related to fewer cancer-related health worries (beta=-0.22). Having more symptoms during treatment is also a predictor of having more cancer-related health worries (beta=0.20). The most consistent predictor of psychosocial distress is dispositional optimism/pessimism, with more optimistic individuals reporting fewer cancer-related health worries (beta=-0.27), lower levels of both anxiety (beta=-0.16) and depression (beta=-0.23). Overall, for many older adult, long term survivors, the legacy of cancer continues in terms of cancer-related health worries. In spite of these, for most survivors, their quality of life is not dramatically compromised either physically or psychologically. PMID- 16041842 TI - The pharmacokinetics of a novel anti-tumor agent, beta-elemene, in Sprague-Dawley rats. AB - beta-Elemene, a natural sesquiterpene, is a novel anti-tumor agent. The pharmacokinetics of beta-elemene after single i.v. administration have been investigated in male SD rats. beta-Elemene was administered at three doses (50, 75 and 100 mg/kg) and a full pharmacokinetic profile was obtained. beta-Elemene was metabolized extensively and eliminated rapidly. High permeability of beta elemene through the blood-brain barrier was found following i.v. administration based on the brain/plasma ratio. Following i.v. administration, the drug was eliminated primarily as metabolite and minimally as unchanged drug. Cumulative fecal, biliary and urinary excretion of beta-elemene in rats was 0.61%, 0.06% and 0.003% of the administered dose (75 mg/kg) at 32 h after administration, respectively. These results indicate that biotransformation may be the main elimination passage of beta-elemene. The metabolism of beta-elemene was extensive and the structure of the metabolite (M1) in rat bile was determined by GS/MS and NMR analysis. PMID- 16041843 TI - Endocrine-related resources from the National Institutes of Health. PMID- 16041845 TI - The NIH policy on enhancing public access to publications resulting from NIH funded research:can we streamline the process for our authors? PMID- 16041846 TI - CPR: are we doing it wrong? Some say just doing chest compressions might be easier and more reliable. And there are questions about where the defibrillator fits in. PMID- 16041848 TI - Dieting with a French accent. It's not true that French women don't get fat, but the best-selling diet book is fun and, who knows, may help some people lose a few pounds. PMID- 16041847 TI - Nicotine: it may have a good side. It gets people hooked on cigarettes, but researchers hope that nicotine and related compounds will have therapeutic uses. PMID- 16041849 TI - Should you be taking aspirin? It depends. PMID- 16041850 TI - By the way, doctor. I'm confused. I've read your articles about heart disease being caused by inflammation. If the COX-2 drugs are anti-inflammatory, why do they seem to make heart disease worse, not better? PMID- 16041852 TI - Medicaid prescription limit could hurt HIV patients. PMID- 16041851 TI - Global. WHO: access to safe blood is limited. PMID- 16041853 TI - Supreme Court. State laws will not protect medical marijuana users. PMID- 16041854 TI - HIV & drugs. Meth use develops stronger link to HIV risk. PMID- 16041855 TI - Limitations period trips up needlestick injury claim. PMID- 16041857 TI - Activism. Activists' claim of false arrest survives dismissal. PMID- 16041856 TI - Officers get immunity after disclosing inmate's serostatus. PMID- 16041858 TI - Ethics. AIDS drug experiments on foster children violated rules. PMID- 16041859 TI - [Diabetes mellitus in childhood and adolescence. 1. Specific management by a multidisciplinary team]. AB - The management and follow up of diabetes in youth is a multidisciplinary challenge due to both short and long term objectives. Awareness of the feelings and problems faced by the families is critical. The experience of our team has started in the 1960s and is briefly described and updated in this article. PMID- 16041860 TI - [Diabetic nephropathy]. AB - Diabetic nephropathy is a constantly increasing pathology in western countries. The trend is more pronounced in type 2 diabetic patients than in type 1 diabetic patients. Among individuals with type 2 diabetes, kidney disease is often multifactorial. This paper reviews recent developments in the pathophysiology, epidemiology and treatment of diabetic nephropathy. PMID- 16041861 TI - [Hypertension and diabetes]. AB - Hypertension frequently accompanies diabetes mellitus, as it is present in 50% of diabetic patients. Hypertension can sometimes preceed diabetes. In type 2 diabetes, insulin resistance plays a major role in the hypertensive risk. In type 1 diabetes, nephropathy is often noted as soon as hypertension is present. Both hypertension and diabetes increase the risk for cardiovascular and renal complications. For their prevention, first of all, modification of the diet with increasing exercise must be proposed, associated to antihypertensive agents with a blood pressure target lower than 130/80 mmHg. Renin-angiotensin blockers constitute the main drug therapy in such patients associated with diuretics or betablocker if angina pectoris is present or even calcium channel blocker when large arteries abnormalities exist. A frequent evaluation of the cardiovascular risk is required together with research of renal dysfunction or microproteinuria. PMID- 16041862 TI - Pectus excavatum surgically repaired using sternum realignment and splint techniques in a young cat. AB - Successful surgical repair of pectus excavatum deformities has so far only been reported in very young animals. This case report describes an alternative technique for repairing pectus excavatum in a young cat. The cat had shown moderate deformity with slight respiratory impairment when young and had experienced clinical problems with age. A sternum realignment technique involving a pin associated with an external splint was used. This alternative technique was a safe and efficient procedure in the five-month-old cat. PMID- 16041863 TI - Effect of deguelin on UVB-induced skin carcinogenesis. AB - Nonmelanoma skin cancer afflicts more than one million people in the U.S. annually, highlighting the need for more effective preventive regimens. We have investigated the ability of deguelin, a plant-derived rotenoid with cancer chemopreventive activity, to inhibit UVB-induced skin carcinogenesis with the SKh 1 mouse model. Topically-applied deguelin significantly inhibited the multiplicity of UVB-induced skin tumors, indicating potential as a human skin cancer chemopreventive agent. Mechanistic studies to determine the potential of deguelin to block a number of established UVB-induced molecular events yielded negative results [including UVB-induced AP-1 DNA binding, c-fos and TNFalpha mRNA induction, arachidonic acid release and UVB-induced phosphorylation of mTOR (Ser2448), akt (Ser473) and erk (Thr202/Tyr204)]. These results are of interest as they contradict a major hypothesis for the mode of action of deguelin, i.e., a general down regulation of signal transduction based on inhibition of NADH dehydrogenase and depletion of ATP levels. In the current work, however, deguelin was found to activate 5' AMP-activated kinase (AMPK), a protein that acts as a cellular energy sensor. This is the first report of a chemopreventive agent having this effect and suggests a possible role for AMPK in cancer chemoprevention. PMID- 16041864 TI - Reassessing carbamazepine in the treatment of bipolar disorder: clinical implications of new data. AB - This monograph summarizes the proceedings of a roundtable meeting convened to discuss the role of carbamazepine in the treatment of bipolar disorder, in light of new data and the recent indication of carbamazepine extended-release capsules (CBZ ERC) for use in the treatment of acute manic and mixed episodes. Two lectures were presented, followed by a panel discussion among all 6 participants. A summary of the two pivotal trials of CBZ ERC and their pooled data along with other relevant data is presented first. Next, historical trends of carbamazepine and the agent's use in acute mania, bipolar depression, and maintenance are reviewed, emphasizing clinical implications of efficacy, safety, tolerability, and drug interactions. Finally, the panel discussion provides recommendations for the use of carbamazepine in different phases of the illness, taking into account adverse effects and drug-drug interactions. Panel discussants agree that current data confirm the utility of CBZ ERC as an effective treatment for acute manic and mixed episodes in bipolar disorder. Carbamazepine may also prove to be an option for maintenance treatment. Tolerability of the drug is related to dose and titration, and overall safety limitations regarding carbamazepine usage are comparable to other medications. For some patients, the main challenges to use of carbamazepine may be common drug-drug interactions and increased side effects related to aggressive introduction during treatment of acute manic and mixed episodes. Thus, carbamazepine may be a lower priority option for patients who are taking multiple medications, such as elderly individuals with medical comorbidity, due to the potential for drug interactions. Important benefits of carbamazepine include the low propensity toward weight gain and evidence of good tolerability with long-term treatment. (At present there are no available data from long-term, placebo-controlled studies evaluating the effects of carbamazepine or CBZ ERC on weight.) Thus, carbamazepine may be a good option for patients who are concerned about weight gain or who are intolerant of or respond poorly to other medications. Further efforts are needed to update physicians on the use of carbamazepine relative to other medications in the treatment of bipolar disorder. PMID- 16041865 TI - Motion contrast classification is a linearly nonseparable problem. AB - Sensitivity to image motion contrast, that is, the relative motion between different parts of the visual field, is a common and computationally important property of many neurons in the visual pathways of vertebrates. Here we illustrate that, as a classification problem, motion contrast detection is linearly nonseparable. In order to do so, we prove a theorem stating a sufficient condition for linear nonseparability. We argue that nonlinear combinations of local measurements of velocity at different locations and times are needed in order to solve the motion contrast problem. PMID- 16041866 TI - Edgeworth-expanded gaussian mixture density modeling. AB - Instead of increasing the order of the Edgeworth expansion of a single gaussian kernel, we suggest using mixtures of Edgeworth-expanded gaussian kernels of moderate order. We introduce a simple closed-form solution for estimating the kernel parameters based on weighted moment matching. Furthermore, we formulate the extension to the multivariate case, which is not always feasible with algebraic density approximation procedures. PMID- 16041867 TI - Predicting stimulant medication response in ADHD: evidence from an integrated profile of neuropsychological, psychophysiological and clinical factors. AB - There have been significant advances in understanding the neurobiology of Attention-Deficit/Hyperactivity Disorder (ADHD) and it is timely to examine the ability of biological and psychological markers to predict medication response in this disorder. We evaluated prediction of medication response in adolescent ADHD using neuropsychological testing and psychophysiological measures of central and autonomic function. Fifty ADHD adolescents participated in pre- and post stimulant medication testing. Separately ranked performance in auditory oddball and visual Working Memory (WM) tasks determined 20 "responders" and 20 "non responders" with 10 "neutrals" excluded from the discriminant function analyses (DFA). For both oddball and WM performance rankings, the two groups did not differ in age, sex, or handedness. However, responders did have higher levels of symptomatology than non-responders at baseline. Pre-stimulant medication psychophysiology variables were used as predictors in each DFA. Oddball performance correctly classified 85.0% of responders and 95.0% of non-responders. Better response was associated with increased resting beta power (left posteriorly), delayed oddball target N1 (frontally), decreased oddball target P2 (left posteriorly) and decreased WM distractor P3 (right frontally). Working memory performance classified 80.0% of responders and 90.0% of non-responders, with a broadly similar profile of psychophysiological predictors. These finding indicate the value of integrating neuropsychological and psychophysiological data in predicting medication response in ADHD. PMID- 16041868 TI - Immune-based therapy shows promise. PMID- 16041869 TI - Rare strain update in New York. PMID- 16041870 TI - The insulin-like growth factor system in cancer prevention: potential of dietary intervention strategies. AB - The insulin-like growth factor (IGF) system is related to proliferation and tumor growth, and high levels of circulating IGF-I are thought to be a risk factor for several types of cancer. This review summarizes the epidemiologic evidence for an association between circulating IGF-I and cancer risk as well as the experimental evidence for a causal relation between the endocrine IGF system and tumor growth. The potential for dietary intervention to alter the IGF system and thereby cancer risk is supported by several lines of evidence. Postulated mechanisms of action are as follows: (a) reduction of levels of circulating IGF-I, which will decrease activation of the IGF-I receptor and subsequent signaling pathways; (b) increasing local IGF binding proteins, which may have IGF-dependent effects through obstruction of IGF interaction with local IGF-I receptor as well as IGF independent effects; and (c) interference with estrogens and estrogen receptor action, which may have direct (and possibly synergistic) effects on IGF signaling. An overview is given of the epidemiologic studies on dietary determinants of circulating IGF-I. Examples of dietary factors, such as dairy protein, lycopene, and phytoestrogens, are used to illustrate the potential mode of action of dietary interventions that may act on the IGF system. In conclusion, the IGF system has every potential to serve as an intermediate for cancer (chemo)prevention studies. On the short term, more research initiatives aimed at the effects of specific food components or dietary strategies on the IGF system both in animal models and in humans are warranted. PMID- 16041871 TI - 2005 resource guide. PMID- 16041872 TI - New glucocorticoids on the horizon: repress, don't activate! AB - Glucocorticoids are highly effective drugs; their immunosuppressive and antiinflammatory actions are used in the treatment of many rheumatic and other inflammatory diseases. However, their use is sometimes considerably limited by numerous adverse reactions. For this reason, great efforts have been made in recent years to develop innovative glucocorticoids or glucocorticoid receptor ligands that have improved therapeutic effect/adverse reaction ratio. We summarize the position and critically discuss the following products that are currently under development: (1) selective glucocorticoid receptor agonists (SEGRA or dissociating glucocorticoids); (2) nitrosteroids; and (3) long circulating liposomal glucocorticoids. Finally, we describe the state of research on membrane-bound glucocorticoid receptors as possible further targets for specific glucocorticoid actions. PMID- 16041873 TI - Borne for another. PMID- 16041874 TI - Legislating advance directives for the terminally ill: the living will and durable power of attorney. PMID- 16041875 TI - Involuntary sterilization of mentally retarded minors in Nebraska. PMID- 16041876 TI - Abortion--child capable of being born alive. Rance v. Mid-Downs Health Authority. PMID- 16041877 TI - Assisted conception and clinical practice: whose freedom is it? PMID- 16041878 TI - Child--medical treatment--court's inherent jurisdiction--treatment contrary to clinical judgment: Re J (a minor)(medical treatment). PMID- 16041879 TI - The Irish abortion controversy. PMID- 16041880 TI - AIDS, prisoners, and the law. PMID- 16041881 TI - The right to found a family. PMID- 16041882 TI - The reluctant survivor. PMID- 16041883 TI - The reluctant survivor. PMID- 16041884 TI - In vitro veritas. PMID- 16041885 TI - T.B. or not T.B.? PMID- 16041886 TI - [The 21st world congress of the International Association of Angiologists: review of the materials]. PMID- 16041887 TI - Soviet Union admits to abuses of psychiatry. PMID- 16041888 TI - Fixation of mast cells. PMID- 16041889 TI - How much do doctors use quality of life information in primary care? Testing the trans-theoretical model of behaviour change. AB - This study aimed to find out whether General Practitioners (GPs) use quality of life (QOL) information in primary care, to explore their reasoning and to assess any barriers to use. A second purpose was to see whether the Trans-Theoretical Model (TTM) of behaviour change could be applied to doctor's use of QOL information in primary care. A representative, cross-sectional sample of 800 GPs was approached in a national postal survey; 280 (38%) provided qualitative and quantitative information. Most GPs said that QOL was interesting and important. Users had seen more information and scales, and were more aware of its uses; only 8% had ever used formal standardised questionnaires. The main barriers to implementation were a shortage of time and information, and experience with using QOL assessment. A sizable minority wanted to know more. Seventy-one percent would use QOL to monitor treatment effectiveness. The five stages of the TTM were used to identify whether GP's were predisposed to use QOL information from their knowledge levels and values. While there was some support for the model, the TTM did not sensitively and reliably discriminate between users and non-users on all variables, and so has limited value here. PMID- 16041890 TI - Assessing a conceptual framework of health-related quality of life in a HIV/AIDS population. AB - With the recognition of health-related quality of life (HRQOL) as an important outcome in the course of HIV-disease, it is important to gain a better understanding of the complex relationships among the various factors that influence it. This study assesses a conceptual framework of HRQOL, consisting of disease status, socio-economic status (SES), behavioral variables, symptom status, role-specific functional status and HRQOL, among a multiethnic, economically disadvantaged population of individuals living with HIV/AIDS. Self report data were collected from 348 patients receiving care at a large HIV/AIDS care center, serving residents of a large metropolitan area. The relationships between the study variables were examined using structural equation modeling. Results indicated that the hypothesized framework provided a well-fitted solution to the data, chi2(44df) = 57.62], p = 0.08 and root mean square error of approximation = 0.03, 90% confidence interval 0.01; 0.05. This framework suggests that health-related variables fall along a continuum, beginning with disease status and ending in generic HRQOL. In addition, the framework suggests that behavioral factors (i.e., smoking status, alcohol consumption, and illicit drug use) and SES exert significant effects along this continuum and should be carefully considered when analyzing and interpreting HRQOL data. PMID- 16041891 TI - Nature and correlates of SF-12 physical and mental quality of life components among low-income HIV adults using an HIV service center. AB - OBJECTIVES: This study describes health-related quality of life (HRQOL) among low income HIV adults using an HIV service center, compares participants' scores to US published norms for the general population and persons with chronic conditions, and examines relationships between patient characteristics, nonadherence, and HRQOL. METHODS: A cross-sectional survey of individuals on antiretroviral therapy was conducted. The Medical Outcomes Study SF-12 was used to assess HRQOL. Medication nonadherence was assessed using the 9-item Morisky Adherence Scale. Data also were collected on social support, CD4 cell count, and time since diagnosis. RESULTS: Approximately 84% of the 86 participants were male, 50% were white, and 39% were black. The mean +/- SD Physical component summary (PCS-12) score of 41.0 +/- 12.5 and Mental component summary (MCS-12) score of 41.9 +/- 11.0 were lower than US general population norms (p < 0.001). PCS-12 scores were similar to those of patients with other chronic conditions. Respondents reported lower MCS-12 scores than patients with hypertension and diabetes (p < 0.006). Employment and higher social support had positive associations with PCS-12 scores and nonadherence had a negative association with MCS-12 scores (p < 0.05). CONCLUSIONS: HRQOL in this sample of low-income HIV adults was comparable to other HIV populations. Identifying strategies for increasing social support and medication adherence for economically disadvantaged persons with HIV/ AIDS may improve their HRQOL. PMID- 16041892 TI - Abuse, HIV status and health-related quality of life among a sample of HIV positive and HIV negative low income women. AB - The assessment of a person's quality of life as it relates to health, HIV status and intimate partner violence (IPV) among women has been limited in its scope of investigation. Consequently, little is known about the adjusted and combined effects of IPV and HIV on women's health status and QOL. 445 women (188 HIV + 257 HIV -) residing in an urban low income area were interviewed regarding current IPV experiences (no IPV, IPV more than 1 year ago, IPV in last year), HIV status (positive and negative), use of illicit drugs, and presence of instrumental social support. Health-related QOL (HRQOL) was measured using the MOS-HIV. Stratified bivariate analyses demonstrate that living with HIV or having experienced IPV in the past year was significantly associated with poorer levels of HRQOL. Multiple logistic regression models indicate a robust negative relationship between the experience of IPV in the past year, living with HIV, use of illicit drugs and a protective effect of social support on women's reported HRQOL. The results of the bivariate and multivariate analyses provide evidence that there are independent and adjusted detrimental associations of the experience of IPV and living with HIV with women's HRQOL. As HRQOL is a good indicator of physical and mental health, these findings should alert health care and other service providers to their responsibility to screen and treat women experiencing intimate partner violence and living with HIV. PMID- 16041893 TI - Sleep quality and health-related quality of life in HIV-infected African-American women of childbearing age. AB - A descriptive, correlational design was used to examine the associations of sleep quality and stage of illness with health-related quality of life (HRQOL) in HIV infected African-American women. Participants were recruited from 12 health clinics and AIDS service organizations (ASO) in Georgia, North Carolina, and South Carolina. The sample consisted of 144 African-American women who ranged in age from 20 to 48 years (m = 34.8, SD = 6.8). The Pittsburgh Sleep Quality Index (PSQI) and the Medical Outcomes Short-Form Health Survey (SF-36) were administered. Participants were categorized as good sleepers (PSQI global score < 7) or poor sleepers (PSQI global score > or = 7) using the median global sleep quality score. Differences in HRQOL between good and poor sleepers, as measured by the SF-36, were tested using MANOVA. Good sleepers scored significantly higher (p < 0.0001) for each SF-36 quality of life dimension and the mental and physical health summary scores. Multiple regression analysis indicated that sleep quality is associated with HRQOL, independent of the individual's stage of illness, more so with mental HRQOL than with physical HRQOL. The results suggest that treatment for poor sleep quality should be a primary concern for the treatment of HIV infection and a means for improving HRQOL. PMID- 16041894 TI - Comparison of preference-based utilities of the 15D, EQ-5D and SF-6D in patients with HIV/AIDS. AB - OBJECTIVE: This article compares preference-based utilities from the multiattribute utility instrument 15D with those derived from the EQ-5D and the Short Form 36 (SF-6D) in patients with HIV/AIDS. In particular, we wanted to examine if the finer descriptive system of the 15D would result in better discriminative capacity or responsiveness. METHODS: In a prospective observational study of 60 Norwegian patients with HIV/AIDS from two hospitals, the authors compared scores, assessed associations with disease staging systems, and assessed test-retest reliability and responsiveness of the instruments. RESULTS: On average, the 15D gave higher utility scores than the other two measures, the mean utility scores were: 15D--0.86, SF-6D--0.73, and EQ-5D Index- 0.77. Test-retest reliability was acceptable for all measures, with intraclass correlation coefficients between 0.78 and 0.94. The correlation between scores of the 3 scales was substantial (p = 0.74-0.80). There was no major difference in responsiveness between the measures. CONCLUSIONS: The different measures gave different utility values in this sample of patients with HIV/AIDS, although many of the measurement properties were similar. There was no evidence for better discriminative capacity or responsiveness for the 15D, than for the two other multiattribute measures. PMID- 16041895 TI - The relationship between lipodystrophy-associated body changes and measures of quality of life and mental health for HIV-positive adults. AB - OBJECTIVES: To investigate the relationship between lipodystrophy-specific symptom severity and wellbeing. METHODS: HIV-positive adult patients with body fat redistribution (lipodystrophy syndrome) associated with antiretroviral therapy reported their total non-lipodystrophy symptoms and side effects and completed measures assessing body fat changes (yielding Atrophy, Hypertrophy and Total Lipodystrophy scores), mental health and quality of life. Effects of total symptom complex and lipodystrophy severity on quality of life and mental health were analyzed using Spearman's rho correlations. Logistic regression analyses were utilized to determine the relative-odds of depression produced by overall symptom count and lipodystrophy score increments. RESULTS: Mean ratings for Hypertrophy and Atrophy corresponded to 'very mild' and 'mild' degrees of severity, respectively. The total symptom complex was associated with ratings for most of the mental health and quality of life measures. Patient-perceived body image scores were the sole study variable responsive to lipodystrophy severity ratings. In comparison to reference norms, a pronounced degree of body image impairment was evident. CONCLUSION: Although responsive to the total symptom profile, psychosocial measures typically utilized for evaluating quality of life and mental health status in HIV disease lacked sensitivity and specificity for measuring the consequences of lipodystrophy-associated fat distribution changes alone. Lipodystrophy severity did impact negatively on body image. PMID- 16041896 TI - Improvements in health-related quality of life following a group intervention for coping with AIDS-bereavement among HIV-infected men and women. AB - BACKGROUND: AIDS-related bereavement is a severe life stressor that may be particularly distressing to persons themselves infected with HIV. Increasing evidence suggests that psychological health is associated with disease progression, HIV-related symptoms, and mortality. PURPOSE: This study assessed change in health-related quality of life among HIV + persons following a group intervention for coping with AIDS-related loss. METHODS: The sample included 235 HIV + men and women of diverse ethnicities and sexual orientations who had experienced an AIDS-related loss within the previous 2 years. Participants were randomly assigned to a 12-week cognitive-behavioral bereavement coping group intervention or offered individual psychotherapy upon request. Quality of life was assessed at baseline and 2 weeks after the intervention. RESULTS: Participants in the group intervention demonstrated improvements in general health-related and HIV-specific quality of life, while those in the comparison remained the same or deteriorated. Effect sizes indicated that the majority of change occurred in women. CONCLUSION: This bereavement group aimed at improving coping with grief also had a positive impact on health-related quality of life among HIV + men and women, and suggests that cognitive-behavioral interventions may have a broad impact on both emotional and physical health. PMID- 16041897 TI - Assessing quality of life in adult cancer survivors (QLACS). AB - This article describes development of a quality of life measure designed to assess issues relevant to long-term cancer survivors. In-depth semi-structured interviews were conducted with 58 long-term cancer survivors to identify domains most relevant to long-term survivors (> or = 5 years post-diagnosis). Self-report items were developed from these interviews and administered to a second sample of 242 long-term survivors. Domains and items were selected from the item pool by a combination of factor analysis and criterion-based item selection. Five cancer specific domains were identified (appearance concerns, financial problems, distress over recurrence, family-related distress, and benefits of cancer) along with seven generic QOL domains (negative feelings, positive feelings, cognitive problems, sexual problems, physical pain, fatigue, and social avoidance). Cronbach's alpha was 0.72 or greater for each domain. Correlations between domain scores and criterion measures were 0.72 or higher in all but one generic domain (social avoidance), but somewhat lower on cancer-specific domains. The new multidimensional measure has good internal consistency and validity and is appropriate for comparisons between cancer and non-cancer populations, as well as long-term follow-up of cancer patients. PMID- 16041898 TI - Proxy ratings of health related quality of life in patients with hepatocellular carcinoma. AB - The present study prospectively assessed consistency of ratings of health-related quality of life between patients with hepatocellular carcinoma and two types of proxy raters, family caregivers and oncologist care providers. Patients and proxies completed the Functional Assessment of Cancer Therapy-Hepatobiliary (FACT Hep) at baseline prior to treatment (82 patients plus proxies), at 3-months (32 patients plus proxies), and 6-months follow up (16 patients plus proxies) after diagnosis. Patient and proxy demographic data and patient medical data were also collected. At baseline, significant intra-class correlations (ICC) were found for patient--caregiver ratings of physical, social/family, and functional well-being, additional concerns (symptoms, side effects), and overall health related quality of life (HRQL); for patient--care provider ratings of physical well-being; and for caregiver--care provider ratings of physical and emotional well-being. At 3 months follow-up, the most significant number of ICCs were recorded across all three rater pairs. Significant consistencies were found for overall HRQL, and all subscale scores except for social and family well-being. The fewest significant ICCs were found at the 6-month follow-up, and were for patient--caregiver ratings of physical and functional well-being, and additional concerns; and for caregiver -care provider ratings on overall HRQL. These results suggest that family caregivers are adequate proxies of patient physical and functional well-being and additional concerns throughout the patient's illness, more so than oncologist care providers. Further research is warranted with larger samples. PMID- 16041899 TI - Validation of the care notebook for measuring physical, mental and life well being of patients with cancer. AB - To measure patients' QOL in the daily practice of clinical oncology, we developed and tested the Care Notebook. This instrument has 24 questions expressed in single words or short phrases to make it more acceptable to patients. The Care Notebook, EORTC QLQ-C30 and FACIT-Sp-12 were administered to 249 outpatients with cancer. Construct validity was investigated by cluster analysis and multitrait scaling analysis. The results showed that three scales (physical well-being, mental well-being, and life well-being) could explain 55% of the variance in scores. The life well-being scale could be divided into subscales of Daily Functioning, Social Functioning, and Subjective QOL. Multitrait scaling analysis confirmed convergent and discriminant validity of these scales and subscales. Internal consistency and test-retest reliability were favorable. Differences in Care Notebook scores were also consistent with differences in performance status rating (known-groups validity), and Care Notebook scores correlated with EORTC QLQ-C30 and FACT-Sp-12 scores (concurrent validity). The Care Notebook allows clinical oncologists to easily collect valid and reliable QOL information of physical, mental, and life well-being repeatedly and with minimal burden on patients. PMID- 16041900 TI - Proxy assessment of quality of life in pediatric clinical trials: application of the Health Utilities Index 3. AB - BACKGROUND: With increased cure rates, pediatric oncology protocols increasingly seek to document the impact of treatment on patients' disease, symptoms, and functional capacity. PROCEDURE: Nurses as proxy respondents used the Health Utilities Index 3 (HUI3) to assess the health-related quality of life (HRQL) in twenty-five patients (age 6 years or older) enrolled on a frontline protocol for leukemia. HRQL observations (n = 70) were made at three different time points to coincide with high-dose methotrexate therapy. Additionally, the proxy respondents evaluated the ease of use of the instrument and the data quality. RESULTS: As patients' health status declined, the number of unassessable HRQL items increased. These missing data made scoring cumbersome and precluded calculation of the overall HRQL scores for nearly 50% of the patients. CONCLUSIONS: Use of the provider proxy-assessed HUI3 in pediatric cancer trials may result in a high proportion of missing data. Trials may benefit more from the use of HRQL measures that consider the acuity of the child's illness, domains specific and sensitive to both disease and treatment, and items that can be proxy-assessed independent of input from parent or patient. Evaluations that combine child self-reports with both parent and provider reports may ultimately provide the most reliable and comprehensive perspective on children's quality of life. PMID- 16041901 TI - Quality of life profile--adolescent version: assessing the relationship of covariates to scale scores using structural equation modeling. AB - This study investigates the effects of covariates on adolescent quality of life (QOL) using structural equation modeling (SEM). The Quality of Life Profile- Adolescent Version, a generic 54 item self-reported questionnaire, was administered to 363 adolescents aged between 10 and 18 years who were enrolled in 20 secondary schools within metropolitan Perth, the capital of Western Australia. Previous results based on a second-order SEM provided preliminary support for measuring adolescent QOL in terms of physical health, psychological, social, environment and opportunities for growth and development. The second-order model is further enhanced in this study by incorporating covariates age, presence of a chronic condition, amount of control and perception of health. The amount of control the adolescent perceived had a significant positive impact on QOL, whereas poorer reports of health had a negative impact. However, the presence of a chronic condition was found to have little effect on QOL, while the modest inverse relationship between age and QOL was not statistically significant. PMID- 16041902 TI - Development and pilot-testing of a health-related quality of life chronic generic module for children and adolescents with chronic health conditions: a European perspective. AB - Health-related quality of life (HRQOL) assessment in children and adolescents with chronic health conditions is increasingly considered as a relevant topic. The aim of the EU-funded DISABKIDS project is to develop, test, and implement European instruments for the assessment of HRQOL of children and adolescents with disabilities and their families. The current paper describes the development and pilot testing of a chronic generic HRQOL measure. Using literature searches, expert consulting and focus groups with children/adolescents and their families, items of the instruments were developed and translated into the respective languages. A pilot test with 360 children and adolescents was conducted. Children and adolescents (8-12, 13-16 years) with different chronic health conditions (asthma, epilepsy, diabetes, arthritis, atopic dermatitis, cerebral palsy, and cystic fibrosis) as well as their families were included. Data were analysed according to predefined psychometric and content criteria. Psychometric analyses resulted in a 56-item chronic generic HRQOL questionnaire with six domains ('Medication', 'Physical', 'Emotion', 'Independence', 'Social Inclusion', 'Social Exclusion') with acceptable internal consistency. PMID- 16041903 TI - Health-related quality of life of mothers of children with leukemia in Japan. AB - OBJECTIVE: The study compared the health-related quality of life (HRQOL) of Japanese mothers of children with leukemia to that of mothers of children without leukemia. METHOD: We used the Medical Outcomes Study 36-item Short-Form (SF-36) to measure the HRQOL of 97 mothers of children (average age 6.2, range 0-14) with leukemia diagnosed between 1999 and 2000, and compared their scores to those of 240 mothers of children without leukemia matched to the children with leukemia. MAIN RESULTS: Of the eight unadjusted domain scores of the SF-36, five were significantly low among mothers of children with leukemia compared to mothers of children without leukemia: 'role-physical functioning' (RP), 'general health perception' (GH), 'vitality' (VT), 'social functioning' (SF) and 'mental health' (MH). Their SF-36 domain scores, when adjusted for demographic and clinical factors were also significantly low in RP, GH, VT, SF, RE and MH. The MH and SF scores in mothers of children with leukemia requiring hospital care were, respectively, approximately 20 points (1 standard deviation (SD), p < 0.0001) and 30 points (1.5 SD, p < 0.0001) lower than that of mothers of children without leukemia. CONCLUSION: Mothers of children with leukemia requiring hospital care have poor HRQOL, particularly with regard to mental health and social functioning, and are at a greater risk for depression. These results suggest that the current system for treating leukemic diseases of children in Japan should also include close monitoring of mothers' mental health, and provision of appropriate treatment and psycho-social support. PMID- 16041904 TI - Health state preference scores of children with spina bifida and their caregivers. AB - Cost-effectiveness evaluations of interventions to prevent or treat spina bifida require quality of life information measured as preference scores. Preference scores of caregivers also may be relevant. This study tested whether the preference scores of children with spina bifida and their caregivers would decrease as disability in the child increased. Families of children aged 0-17 with spina bifida (N = 98) were identified using a birth defect surveillance system in the state of Arkansas. Primary caregivers of children with spina bifida identified other families with an unaffected child (N = 49). Preference scores for child health states were determined using the Health Utilities Index--Mark 2 (HUI2). Caregiver preference scores were determined using the Quality of Well Being (QWB) scale. Children with spina bifida were categorized into three disability levels according to the location of the child's lesion. Mean preference scores declined for both affected children and the primary caregiver as disability in the child increased. In multivariate analysis, the preference score of the child was a significant and positive predictor of the primary caregiver's preference score. A more modest association was found for caregiver health preference scores by lesion location. The findings can inform cost effectiveness evaluations of interventions to treat or prevent spina bifida. PMID- 16041905 TI - EQ-5D in a general population survey--a description of the most commonly reported EQ-5D health states using the SF-36. AB - The importance of studying health-related quality of life in the general population has increasingly been emphasized. From a public health perspective, this benefits the identification of population inequalities in health status. One of the currently most popular instruments is the EQ-5D. Evaluations of the EQ-5D generally focus on the overall preference-based index. As this index has a built in value, exploration of the information from the underlying health states is also important. In this study, the ten most commonly reported EQ-5D health states are described using the SF-36. Data collected in 1999 by questionnaires mailed to a random sample aged 20-74 in south-eastern Sweden were used (n = 9489). Almost 43% reported the best possible EQ-5D health state and 78% were accounted for by three EQ-5D health states. The EQ-5D health state classification was largely reflected by the SF-36, with the EQ-5D items mobility, usual activities, pain/discomfort and anxiety/depression tapping most clearly on the SF-36 scales physical functioning, role limitations due to physical health problems, bodily pain, and mental health, respectively. However, within the same level of EQ-5D (i.e., moderate problems) there was a rather large variation of SF-36 scale scores, particularly regarding the EQ-5D item pain/discomfort and the SF-36 scale BP. PMID- 16041906 TI - On loss of activity and independence, adaptation improves life satisfaction in old age--a qualitative study of patients' perceptions. AB - The purpose was to improve the understanding of factors are perceived by elderly people as important for their life satisfaction, during and after rehabilitation. Fifteen persons aged 80-94 years were interviewed while in hospital and on two follow-up occasions after discharge. Assessment of motor function using the General Motor Function assessment scale was used for descriptive purposes. Three themes emerged as important for life satisfaction: activity, independence and adaptation. Activity and independence were considered significant for life satisfaction. Basic activity preferences were related to care of one's own body and to social contacts. Control and influence over help and services were regarded as important. Different strategies for adaptation to the consequences of disease were used: reorganisation, interaction with caregivers, mental adaptation and mental activities (used as pastime and escape). Those with declined motor functions limited their activity preferences. A key finding was that pleasant past memories were actively recalled in an effort to achieve current life satisfaction. This adaptation strategy created a sense of life satisfaction, however with a potential risk for concealing dissatisfaction with conditions that might otherwise be correctable. Strategies for improving life satisfaction among old people in rehabilitation are suggested. PMID- 16041907 TI - Is a family equal to the sum of its parts? Estimating family-level well-being for cost-effectiveness analysis. AB - Cost-effectiveness (CEA) analysis of health interventions focuses primarily on individual-level costs and benefits. However, health interventions intended for individuals often have implications for other members of an individual's family and the overall functioning of the family. While researchers have assessed the effects of health interventions on multiple family members, CEA has not routinely incorporated measures of effectiveness or costs from multiple family members. We could imagine, however, that CEA might over- or under-estimate the effects of health interventions if they are limited to individuals targeted by the intervention. Family-level CEA would consider the well-being of and costs borne by multiple family members in response to an individual-level intervention, and potentially lead to more appropriate resource allocation. Using the health plan as a working frame of reference, we explore conceptual issues related to estimating effectiveness at the level of the family for use in CEA. PMID- 16041908 TI - Is a family equal to the sum of its parts: a comment. PMID- 16041909 TI - The impact of non-traumatic hip and knee disorders on health-related quality of life as measured with the SF-36 or SF-12. A systematic review. AB - OBJECTIVES: The purpose of this review is to summarize the available evidence on the impact of non-traumatic hip or knee disorders on health-related quality of life (HRQL), as measured with the Short Form 36 Health Survey (SF-36) or Short Form 12 Health Survey (SF-12), by comparing this with data from reference populations. METHODS: Studies were identified by an electronic search of the MEDLINE, PsychInfo and Cinahl databases. Studies with the following features were included: study population included patients with non-traumatic hip or knee disorders, the SF-36 or SF-12 was used as an outcome measure and mean scores on these HRQL measures were presented. Using mean HRQL scores from the selected studies and scores from reference populations, z-scores were computed. Pooled estimates were computed for subgroups of studies with similar patients in similar settings. RESULTS: A total of 40 studies met the inclusion criteria. Patients with non-traumatic hip and knee disorders scored up to 2.5 standard deviations (SDs) below reference population values, especially on the physical aspects of HRQL. Social and mental aspects were up to 1 SD below reference population values, especially in patients in clinical settings. CONCLUSIONS: The impact of non-traumatic hip or knee disorders on HRQL is substantial, especially on the physical aspects of HRQL. PMID- 16041910 TI - A model of subjective quality of life for outpatients with schizophrenia and other psychoses. AB - This study investigated predictors of quality of life among persons with schizophrenia and other psychoses. On the basis of previous research, it was hypothesised that objective life circumstances, self-variables, psychopathology, activity level, satisfaction with daily activities, and satisfaction with medical care would be determinants of quality of life. 134 persons were investigated, and the analysis was based on Structural Equation Modelling (SEM). A model with two quality of life variables, General quality of life and Security, fitted the data best. A self-variable, Sense of self, and Satisfaction with daily activities, Psychopathology, and Satisfaction with medical care served as mediators and explained the variation in the quality of life variables. External to the mediators, and related to these, were Activity level, Educational level, and Independent living. In conclusion, a self-variable showed the strongest association with quality of life, but two aspects that should be feasible to influence in mental health care, daily activity and medical care, contributed to the subjects' self-rated quality of life. These results render a somewhat optimistic view on how to accomplish a better self-perceived quality of life among people with severe mental illness. PMID- 16041911 TI - Health-related quality of life and health utility for the institutional elderly in Taiwan. AB - OBJECTIVE: To explore the health-related quality of life (HRQOL) and health utility in an institutional elderly population. METHODS: Four hundred sixty-five elderly persons living in long-term care institutions in Taiwan were interviewed using Taiwan's abbreviated version of the World Health Organization Quality of Life (WHOQOL-BREF), rating scale (RS) and the Time-Trade-Off (TTO) utility measurement. RESULTS: The WHOQOL-BREF showed acceptable internal consistency (alpha range: 0.75-0.80 across domains) and validity. The sexual activity facet had the lowest response rate. Educational level, number of chronic diseases, physical performance, and number of caregivers had significant (p < 0.05) impacts on the domain scores of the WHOQOL-BREF. Physical performance had the strongest impact on the physical domain (R2 = 0.40) and accounted for significant percentages of the variance on the other three domains (R2 = 0.06-0.13). The mean RS score (score 0-100) was 61.3 +/- 16.2 (mean +/- SD). The mean TTO utility (score 0-1) was 0.92 +/- 0.22. CONCLUSIONS: Results indicate that the WHOQOL BREF, excepting the sexual activity item, is useful for evaluating HRQOL of conscious elderly in institutions. The validity of TTO utility for studying the institutionalized elderly needs further evaluation. PMID- 16041912 TI - The European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-c30): validation of English version in Singapore. AB - OBJECTIVE: This study aimed to validate the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30, English version 3.0) in Singaporean cancer patients. METHODS: In a cross-sectional study, a heterogeneous sample of cancer patients (n = 57) self-administered a questionnaire containing the QLQ-C30, the Short Form 36 Health Survey (SF-36) and assessing health and sociodemographic status. Construct validity was assessed by testing a priori hypotheses that QLQ-C30 scales would be moderately or strongly correlated with SF-36 scales measuring similar dimensions of health-related quality of life (HRQoL) and that subjects reporting mild symptoms would have better HRQoL scores than those reporting severe symptoms. Internal consistency reliability was assessed using Cronbach's alpha. RESULTS: Strength of Spearman's correlations between the QLQ-C30 and SF-36 scales assessing similar dimensions of HRQoL ranged from 0.35 to 0.67. Subjects with mild symptoms had better scores than those with severe symptoms for all six QLQ-C30 HRQoL scales (p < 0.05 for five scales, Mann-Whitney U tests). Cronbach's alpha ranged from 0.19 for the cognitive functioning scale to 0.91 for the global QoL scale. CONCLUSION: This study provides preliminary evidence for the validity and reliability of the EORTC QLQ-C30 in English-speaking Singaporean cancer patients. PMID- 16041913 TI - The Chinese Medical Interview Satisfaction Scale-revised (C-MISS-R): development and validation. AB - BACKGROUND: The Medical Interview Satisfaction Scale (MISS) measures patient consultation satisfaction. We validated a translated short-form of the original MISS on Hong Kong Chinese women with breast cancer. METHODS: The four highest scoring MISS Cognitive and Affective sub-scales items were administered in Chinese (C-MISS) to 224 female out patients. Phrasing revisions (C-MISS-R) were tested on 82 women. Random split tested factor structure stability. Convergent, divergent and criterion validation against other instruments was performed on 405 women. RESULTS: Two-factors accounted for 61.7% of variance but factor loadings differed from the original. Phrasing revision increased Affective sub-scale item item correlations exceeding 0.3-64%. Two factors matching those of the original MISS, accounting for 65.2% of variance, explained 36.6% (Cognitive) and 28.8% (Affective) of variance, respectively. Alpha was 0.84,0.74 and 0.83 for the Cognitive and Affective sub-scales and total respectively. Correlations of difficulties with treatment decision making (r = -0.298), Self-efficacy (r = 0.194), optimism (r = 0.33), psychological morbidity (r = -209), marital status, education and age indicated acceptable validity. Test-retest reliability was 0.410. DISCUSSION: The Chinese MISS-Revised (C-MISS-R) has suitable factor structure and psychometric properties for use in consultation studies among Chinese female populations. Further validation is needed for males. PMID- 16041914 TI - Validation of the English version of the KINDL generic children's health-related quality of life instrument for an Asian population--results from a pilot test. AB - OBJECTIVES: To evaluate the psychometric properties of the KINDL questionnaire in an Asian population. METHODS: Consecutive patients with diabetes mellitus (DM) and healthy subjects were recruited to complete the English KINDL questionnaire. The inclusion criteria for patients were age 8-16 years, English-speaking, diagnosed with DM and absence of co-morbid conditions. RESULTS: Thirty children with DM (mean age: 10.7 +/- 1.35 years; 11 M) and 39 healthy subjects (mean age: 10.6 +/- 1.23 years, 17M) completed the child version whereas 31 adolescents with DM (mean age: 14.5 +/- 1.48 years; 15M) and 32 healthy subjects (mean age: 14.3 +/- 0.87 years, 16M) completed the adolescent version. Overall, children with DM reported better HRQoL than healthy children. Although this appeared counter intuitive, several explanations are possible: (1) the development of resilience to the disease over time, (2) our subjects are well-managed, (3) response shift, (4) the provision of high quality medical care, (5) compared to normal children, diabetic subjects and their family pay greater attention to health issues. The reliability coefficients were (overall, scales): KINDL-Kid DM (0.79, 0.44-0.65), KINDL-Kid Healthy (0.71,0.60-0.80), KINDL-Kiddo DM (0.77, 0.37-0.74) and KINDL Kiddo Healthy (0.84, 0.21-0.79). CONCLUSIONS: The KINDL questionnaire appeared promising for use in Asian children. However, further validation in a sample more representative of the general population is required. PMID- 16041915 TI - The management of fatigue in depressed patients. AB - Three quarters of patients who respond to treatment with the newer antidepressants still complain of fatigue. Fatigue is one of the most common and disturbing residual symptoms of depression. Increased serotonin activity in certain areas of the brain contributes to fatigue. It can be counteracted by dopaminergic agents which, interestingly, are enhanced by exercise. Specific steps that can be used to address residual fatigue include cognitive interventions based on those used to address somatoform disorders; graded aerobic exercise; dose reduction or discontinuation of fatigue-inducing antidepressants; and the prescription of such medications as dopaminergic antidepressants (bupropion), stimulants, thyroid preparations, and modafinil. PMID- 16041917 TI - Augmentation of standard antidepressants with atypical antipsychotic agents for treatment-resistant major depressive disorder. AB - Treatment-resistant depression (TRD) is not uncommon; 29% to 46% of patients with depression who are treated with antidepressants fail to respond fully. Dr. Papakostas discusses various augmentation strategies, focusing particular on a possible role for the newer atypical antipsychotics. Their complex neuropharmacological actions suggest such efficacy. One chart review and a number of open-label studies have been encouraging, but 2 double-blind, placebo controlled-studies have offered contradictory results. Considering the side effect profiles of these drugs, a careful risk/benefit assessment is always required. While routinely used for psychotic depression and panic reactions in depressive disorders, their value as augmenters in TRD awaits further study. PMID- 16041916 TI - Genetics and psychopharmacology: prospects for individualized treatment. AB - This article provides a clear and succinct description of the components of inheritance, such as trait transmission, genetic variability, and gene interaction. Genetic sequences constitute the prime focus of pharmacogenetic studies. Variations in drug-metabolizing enzyme systems tend to be monogenic, whereas the pharmacologic effects of medications appear to be polygenic, i.e., complex phenotypes shaped by the interaction of genes and environment. Translated into clinical terms, a history of a good response to a drug in a close relative of a patient is presumed to predict a good response to the same medication by the patient. This seems to hold for antidepressants, antipsychotics, and lithium, but the evidential studies generally have meaningful limitations. Bit by bit, information about the relationship between particular genetic formations and the effectiveness of these medications as well as their side effects, is appearing. The authors cite a number of examples, one such being an association between impaired antidepressant activity and the short allele of SLC6A4. This research promises to strengthen the accuracy, effectiveness, safety, and cost of our psychopharmacological practices. PMID- 16041918 TI - Study designs and outcomes in antidepressant clinical trials. AB - Clinical trials of antidepressants are difficult to design and conduct. In fact, more than half of all recent clinical trials of commonly used antidepressants failed to show statistical superiority for the drug over placebo. This is not necessarily because of the ineffectiveness of the antidepressant, but rather because of an increased response to placebo. Factors that may contribute to these findings remain elusive. Using data from U.S. Food and Drug Administration (FDA) Summary Basis of Approval (SBA) reports and from studies conducted by our group, we reviewed methodological factors used in clinical trials of antidepressants. The 2 most notable factors affecting positive trials are (1) the inclusion of patients with more severe depression, and (2) the use of a flexible-dose design; these may yield results identifying true antidepressant-placebo differences. Severely ill patients with depression respond well to antidepressants but poorly to placebo. Flexible dosing paradoxically reduces the response to placebo without augmenting the response to the antidepressant. All of these findings suggest that the use of placebo is mandatory when assessing new antidepressants. PMID- 16041919 TI - Long-term benefits of early pharmacologic treatment in Alzheimer's disease. Interview by Frederic Flach. PMID- 16041920 TI - [Prevalence of Listeria spp. in dairy farm and evaluation of antibiotic resistance of isolates]. AB - The study was performed to estimate the prevalence and antimicrobial sensitivities of Listeria spp. in raw milk, feaces end environmental samples isolated from 10 dairy in Molise Region. A total of 454 samples were collected, which comprised 40 raw milk, 40 animal faeces and 374 environmental samples. Listeria monocytogenes was never isolated from raw milk specimens; one was isolated from faeces speciments and two were isolated from environmental samples. All isolates were resistant to two or more of antimicrobial agents tested (cephalotin, ampicillin, tetracycline, co-trimoxazole, erytromicin, clindamycin, gentamicin, oxacillin). One isolate of L. monocytogenes was susceptible to all antimicrobial agents tested except oxacillin. This study indicates that faeces, equipments and environment are important reservoirs of Listeria spp. in dairy farm, and can represent potential source of contamination of raw milk. However, the contamination of milk, and the risk of infection, can be effectively eliminated by pasteurisation process. PMID- 16041921 TI - [The influence of professional degree on the knowledge of HIV, HBV and HCV infections in dentistry practice]. AB - The knowledge and preventive practices toward the risk of infection with HIV, HBV and HCV were evaluated in a sample of 254 dentists and odontostomatologies in Palermo, by answering to an anonymous questionnaire sent by mail. Overall, most of the participants showed a good knowledge of the transmission pattern of blood borne viruses as well as good practice for personal hygiene (washing of hands, routine use of gloves, etc.). However 80.8% answered to recap needles after use and more than 40% have had at least one needle injury during the last year in their occupational setting. Moreover, nearly 24% of the interviewed declared to have not made vaccination against Hepatitis B virus infection. Statistical analysis showed a significant correlation between better knowledge, safer practice and different teaching training suggesting that odontostomatology degree might be more appropriate for a better training in preventive medicine for blood borne pathogens. Data also suggest the need of continuous worker education to reduce occupational blood exposures in dentistry. PMID- 16041922 TI - [A vaccination campaign for gypsy children in Rome]. AB - Several studies conducted in Rome have shown low levels of vaccination coverage in gypsy communities. Thus a vaccination campaign targeting to 2400 gypsy children aged 0-13 years, present in 32 settlements in Rome, was conducted in 2002. The campaign was related to vaccinations required and recommended in Italy: diphtheria, tetanus, hepatitis B, pertussis (whooping cough), haemophilus influenzae and measles. In the majority of cases it was decided to carry out the vaccinations directly in the gypsy settlements, in accordance with the methods of pulse immunisation. In the case of small settlements a strategy for reorienting the population to vaccination centres was adopted. Around 2000 children were vaccinated, equivalent to 80% of the paediatric population present during the period. The number of children who have never been vaccinated has decreased from 40% prior to the campaign to 9% after the third week. Vaccination coverage in medium- and small-sized settlements (<200 inhabitants) after the campaign shows values of over 70%; in the large settlements, more modest increases have been recorded and coverage has rarely exceeded 50%. This experience has highlighted the importance of networking between public healthcare institutions and non profit organisations. The mobilisation of a wide range of competences has thus enabled the attainment of a high level of effectiveness. PMID- 16041923 TI - [Epidemiological survey on smoking habit among young students in Sardinia. A cross sectional study]. AB - The study reports the prevalence of cigarette smoking among 11401 high school Sardinian students. The prevalence of smokers (40.2%) significantly differs between gender (41.1% males and 38.4% females). Males have an early initiation of smoking with an evident addictive effect by age. 54.3% are daily smokers and 21.4% smoke 15 or more cigarettes per day. More than 50% smoke to look grown-up and to be accepted by the group. Besides age (OR=1.10; 95%CI: 1.06-1.15), other factors are associated with smoke: low education level of father (OR=1.08; 95%CI: 1.02-1.15), no maternal support (OR = 1.73; 95%CI: 1.17-2.54), to have at least one smoker cohabitant (OR=1.66; 95%CI: 1.54-1.80) and alcohol drinking (OR=3.46; 95%CI: 3.04-3.93). The smokers' knowledge on smoke topics significantly differ from non smokers. Our results suggest the need of community preventive interventions, diversified for specific target populations, to modify the students' behaviours so that they respect their own health and that of their fellow citizens. PMID- 16041924 TI - [Monitoring occupational exposure to volatile anaesthetics in the operating theatre: environmental and biological measurements]. AB - Concentrations of nitrous oxide (N2O) and isoflurane were measured in environmental and urinary samples from subjects occupationally exposed to volatile anaesthetics in operating theatres in a hospital in northern Italy. The aim was to establish whether: an automatic analyzer (Bruel & Kjaer 1302 spectrometer) can be used for fixed position sampling ("anaesthetist zone" and "surgeon/instrument nurse zone"); periodic monitoring of anaesthetics will reduce exposure; exposure to N2O and isoflurane is within legal limits; exposure differs between anaesthetists and surgeons/instrument nurses. Exposure to anaesthetics was monitored twice at six-month intervals. In the first test time spent in the operating theatre was noted and exposure levels were measured automatically. In the second test levels were monitored with passive personal sampling devices. Environmental concentrations of N2O determined by the spectrometer were correlated to urinary levels. Urinary levels of N2O calculated from the regression line were the same as those obtained with the personal samplers. Environmental and urinary levels of N2O decreased significantly from the first to second test. In the second sampling 70% of subjects had levels of exposure to N2O and isoflurane within prescribed environmental limits (50 ppm for N2O and 0.5 ppm for isoflurane). At the first test anaesthetists had significantly higher levels of exposure to N2O than surgeons/instrument nurses. The survey demonstrated that: fixed position sampling data related to time spent in the operating theatre can be used to gauge individual exposure levels; exposure levels decrease after tests following implementation of preventive measures; monitoring needs to be repeated because exposure levels often exceed legal limits; occupational exposure decreases when pollution in the anaesthetic zone is reduced. PMID- 16041925 TI - [Health conditions of immigrant women in Italy]. AB - The number of immigrant women in Italy has increased from 260,000 in 1991 to at least 750,000 in 2003. This article describes the health situation of these women, in particular it deals with reproductive health. Immigrant women are generally young, in good health and they go to the health services mainly for pregnancy, delivery, spontaneous and induced abortion. Forty-eight per cent of acute hospital admissions and 56 per cent of day hospital admissions in 2002 were related to reproduction. Among foreign citizens, the induced abortion rate is three times higher than that reported among Italians, while the risk of spontaneous abortion is similar (97 per thousand and 101 per thousand, respectively). In general, the data show that immigrant women in Italy live in deprived social conditions, which can influence their reproductive choices and their access to health services. In order to take account of their particular needs, it is necessary to modify the health services and plan public health interventions especially for the prevention of induced abortion. PMID- 16041926 TI - [Public food service in Milan city and Hinterland: GMP application (Part 1)]. AB - Food service establishments are recognized as a critical sector concerning foodborne diseases occurrence, that is associated to contributing factors such as the anticipated preparation of meals that are often highly handled, and long-time distributed. A survey has been planned to evaluate the application of HACCP plan, in order to select a statistically representative sample of food services (restaurant, pizza-shop, bar, ..) in two Milan area' Public Health Units (PHU). During the inspections a proper check-list has been filled up in order to give a conformity evaluation about the global situation and about three specific sections: hygiene of food-handlers, procedures control, temperatures management. The food services have been found satisfactory in 9/106 and 5/54 cases in Milan City and in hinterland, respectively; among the two areas, highly significant differences have been revealed about temperatures management (68% and 28% unsatisfactory, respectively). In Milan City restaurants provided with HACCP plan scores are significantly different from unprovided restaurants scores (global and the three sections' evaluation); in Milan hinterland differences between provided and unprovided HACCP plan restaurants regard temperature management scores only. Useful suggestions to improve the quality of surveillance activity come from complex and heterogeneous findings shown in this study. PMID- 16041927 TI - [Public food service in Milan city and Hinterland: food-handlers training level (Part 2)]. AB - The prevention and control of foodborne diseases play an important role in public health and the responsibility of food-handlers is of great concern: their training on food safety is particularly required in the critical sector of public food service establishments. The food-handlers knowledge has been evaluated by a questionnaire filled up in two Public Health Units (PHU) in Milan City and Milan hinterland. Only 11.5% of food-handlers turned at to be good trained; the wrong answers are concentrated on the section titled "temperatures management" (68 and 59% in Milan City and in Milan hinterland, respectively). Only for 31.4% of food handlers the attendance to training courses is well documented, although the knowledge level seems to be partially influenced by training activity: exclusively in Milan City food-handlers an highly significant difference has been found among "trained" and "no-trained" workers in correct answers about temperatures management (p= 0.0046). The frequency of exact answers isn't associated to the satisfaction level revealed during inspection of restaurants. Focusing the attention on the insufficient efficacy of training courses and the lacking connection between knowledge and behaviour, a critical revision of training activity quality seems to be necessary. In future some professional training activities in the field should be promoted. PMID- 16041928 TI - [Domestic injuries in childhood. Results of a survey carried out in a Sicilian area]. AB - A retrospective investigation was performed with an anonymous questionnaire administered to the mothers of 2208 children aged between 3 and 11 years, randomly selected from the residents of three target areas: a country town, a residential and a popular quarter of a chief town (Catania, Sicily). Of the 2208 mothers in the three areas, 1151 completed the questionnaire with an overall response rate of 52.12%. In the last 6 months a large number of children (17.29%) suffered at least one accident at home. Predominating injuries were falls (71.86%) followed by cutting or stab-wounds (13.57%). The more frequent lesions were contusions (42.13%), wounds (29.95%) and fractures (14.22%). Almost the half of children victims of injuries required hospital care (44.72%). The majority of domestic accidents occurred in the bathroom (17.10%), in the kitchen (15.10%) and in the bedroom (10.55%). Fractures and wounds were more frequent among children aged 6-11 and 3-5 years, respectively. The highest frequency of domestic accidents among children of all ages living in the popular quarter (20.89%) and in the country town (19.18%) compared with those living in the residential quarter (11.67%) (chi2= 12,768, P<0.01) could be related to their lower socio economic condition. PMID- 16041929 TI - [What the general practitioner should know about heart failure. Control of pump failure]. PMID- 16041930 TI - [Computerized tomography leads to erroneous diagnostic follow-up. "Stroke" came from the lung]. PMID- 16041931 TI - [Why does the Mediterranean diet work only in the Mediterranean area? "Perhaps foods are spoiled by industry!"]. PMID- 16041932 TI - [Borrelia infection diagnostic challenge. Manage when suspected also without serologic findings!]. PMID- 16041933 TI - [New therapies lower risk of recurrence. Hope for healing in breast carcinoma? (interview by Dr. Judith Neumaier)]. PMID- 16041934 TI - [Diving medicine is more than an inspection of the ear]. PMID- 16041935 TI - [Diving fitness for scuba divers--what the primary care physician should know]. AB - The diving fitness medical examination serves to show and reveal medical conditions that are a contraindication for diving or to evaluate the risk of preexisting conditions. For this reason, it should never have the character of a certification given as a matter of courtesy. Fitness to dive is given if the candidate is healthy and when there are no pathological findings. Even with deviations from the norm, diving is still possible, but with restrictions. Important organ systems for the diving fitness examination are the cardiovascular system, the respiratory organs and the ears. In addition, adequate eyesight is important. The German Society of Diving and Hyperbaric Medicine (GTOUM) has drawn up recommendations on the examination of scuba divers to assist the physician (www.gtuem.org). PMID- 16041936 TI - [Medical aspects of diving in the tropics]. AB - Scuba diving vacations in tropical surroundings belong to the repertoire of most divers. In addition to carefully making travel plans and taking care of the necessary vaccinations and appropriate malaria prophylaxis, the following points also must be observed. The flight itself affects diving safety. In particular, a too short time interval between diving and the return flight can lead to decompression problems. Because most of the diving areas are reached by ship, many divers need a prophylaxis against motion sickness. Moreover, external otitis occurs more frequently while diving in the tropics. Finally, there is potential danger from the sea inhabitants, primarily from scorpion fishes, Portuguese Man of-Wars, box jellyfishes as well as cone snails. PMID- 16041937 TI - [Injuries caused by pressure differences while diving]. AB - Barotraumas are caused by pressure differences. As described by Boyle's Law, barotraumas develop during the descent phase of diving (and much more rarely during the ascent). The most frequently affected are the ears and paranasal sinuses, in addition to the facial skin and eyes. The most important preventive measure is performing pressure compensation in the affected body cavities. Barotrauma is treated symptomatically. PMID- 16041938 TI - [Decompression injuries]. AB - A decompression accident occurs during uncontrolled dive ascent with diving equipment. Through the rapid decrease in the surrounding pressure, gas bubbles form in the blood and tissues. Depending upon the mechanism of onset, the decompression illness (DCI) is classified as decompression sickness (DCS) or arterial gas embolism (AGE). The therapy consists of administering, as quickly as possible, 100% oxygen as well as a volume substitution. The treatment is continued in a recompression chamber. PMID- 16041939 TI - [Problems in the deep: the isopression phase]. AB - Fundamentally, accident mechanisms during the isopression phase of diving are primarily dependent upon the partial pressures of the respiratory gases. An increased nitrogen partial pressure leads to compressed-air intoxication; an increased oxygen partial pressure while diving with oxygen-enriched gas mixtures can trigger an oxygen-induced convulsion. Elevated pCO2 can be provoked by inadequate breathing and/or physical exertion at greater diving depths. Through an adjusted diving behavior and observation of the limits, these problems could be easily avoided. PMID- 16041940 TI - [Blood pressure limits for beginning and goal of antihypertensive therapy (3). What is not proven but plausible?]. PMID- 16041941 TI - [Observational data in elderly pain patients show that fear of opioid therapy is unfounded]. PMID- 16041942 TI - [Heart failure after acute myocardial infarct. Early aldosterone blockade increases survival rate]. PMID- 16041943 TI - [Neurohumoral systems escape control. Heart failure symptoms even once--long-term high risk prognosis]. PMID- 16041944 TI - [Sequelae of health care reform. New retrospective data wave rolls toward physicians]. PMID- 16041946 TI - [Fear of retrospective data initiative. Bavarian physicians stream into the medication program]. PMID- 16041945 TI - [Case values in new EBM. Many general practitioners lose up to 35 percent income]. PMID- 16041947 TI - [Ezetimib and simvastatin. Cooperation at receptors of cholesterol metabolism]. PMID- 16041948 TI - [Goal in hypercholesterolemia: lowering LDL cholesterol level by 50%]. PMID- 16041949 TI - [Meteorism and flatulence. Inflated by wind]. PMID- 16041951 TI - [New paths to highly effective tetracyclines]. PMID- 16041952 TI - [Important research on the controversy over the prion hypothesis]. PMID- 16041953 TI - [The history and status of transplantation medicine]. PMID- 16041954 TI - [Principles of immunosuppression]. PMID- 16041955 TI - [The pharmacology of immunosuppressives]. PMID- 16041956 TI - [Macromolecular immunosuppressive agents]. PMID- 16041958 TI - [Immunomodulators for treatment of rheumatoid arthritis]. PMID- 16041957 TI - [Development of oral and parenteral drug forms]. PMID- 16041959 TI - [Topical calcineurin inhibitors in dermatology]. PMID- 16041960 TI - [Immunosuppressive treatment following kidney transplantation]. PMID- 16041961 TI - [The problem of drug interactions with immunosuppressive agents]. PMID- 16041962 TI - Bloodless open heart surgery: simple and safe. AB - OBJECTIVES: The use of blood or blood products is routine in cardiac surgery, but is associated with various complications. Aware of this, we have always tried to avoid the use of blood products whenever possible. In this study we sought to evaluate the results of this policy. METHODS: The records of 1505 adult patients who underwent coronary (732) or valve (773) surgery under cardiopulmonary bypass (CPB) in 2002 and 2003 were reviewed retrospectively. Of these, 1058 were male (70.3%) and the mean age was 62.1+/-11.4 years. Mean weight was 68.5+/-10.2 kg and body surface area was 1.7+/-0.2 m2, corresponding to a blood volume of 4119.9+/-593.6 ml. Preoperative hematocrit (Hct) was 40.6+/-4.2% and the prothrombin index was 87.0+/-17.4%. A bloodless prime of the bypass circuit was used for all patients with Hct > or =36%. The prime volume was reduced to the minimum possible. Plasma was used when coagulation was deficient. All blood remaining in the CPB circuit was reinfused at the end of the procedure, either in the operating room or in the ICU. Shed mediastinal blood was retransfused in the first 6 hours in the ICU. RESULTS: Operative mortality was 0.7% for coronary and 0.5% for valve patients. Blood or blood products were not used in 77.3% of the patients (88.7% of coronary and 66.5% of valve patients). Blood and/or plasma was initially added to the prime in 18.2% of cases and during CPB in 11%. Hct was 28.9+/-4.0% after initiation and 28.8+/-3.9% after discontinuation of CPB. The number of units (300 cc) of blood used was 0.25.57 per patient (1.09+/-0.73 per patient transfused). The number of units (300 cc) of plasma used was 0.24+/-0.72. Reoperation for bleeding was required in 2.4% of the patients. CONCLUSIONS: This blood-sparing policy is simple, effective and safe, resulting in low mortality and morbidity rates. More than three quarters of the patients did not require blood or blood products. Additional measures are possible to further decrease the use of blood products. PMID- 16041963 TI - Bloodless surgery. PMID- 16041964 TI - Non-invasive coronary artery evaluation with multidetector computed tomography. AB - OBJECTIVES: To evaluate the diagnostic accuracy of 16-slice multidetector spiral computed tomography in the non-invasive detection of coronary artery disease compared to invasive coronary angiography. DESIGN: Prospective study. PATIENTS: We studied 35 patients, 21 men and 14 women, mean age 63.6+/-13.8 years, who underwent elective invasive coronary angiography one week prior to undergoing 16 slice multidetector computed tomography. METHODS: We used a Toshiba Aquilion 16 slice system. Helical acquisition was performed with breath holding after injection of 140 ml of a contrast agent into a peripheral vein. Cross-sectional images were reconstructed with a slice thickness of 0.5 mm or 1 mm. We set 15 anatomical segments for the coronary tree and the evaluation was performed independently for both techniques. Coronary artery lesions with over 50% reduction in diameter were considered significant. Findings from both techniques were evaluated using ROC curve analysis. RESULTS: It was possible to evaluate 87.7% of the coronary tree segments; heavy calcification, enability to perform breath holding, motion artefacts and stents made evaluation of the other segments impossible. Overall sensitivity and specificity were 85.9% and 98.2% respectively with a 95 % confidence interval; positive predictive value was 87% and negative predictive value 97% For the different anatomical segments, sensitivity and specificity were: proximal 88.9% and 97.8%; medial 90.9% and 95.8%; distal 86.7% and 99.1% respectively. CONCLUSIONS: Multidetector computed tomography is a promising non-invasive technique for detecting or ruling out significant coronary lesions. PMID- 16041965 TI - Impact of cardiac rehabilitation on quality of life and depression after acute coronary syndrome. AB - OBJECTIVE: To evaluate the impact of cardiac rehabilitation programs (CRP) on quality of life (QL) and depression at one-year follow-up after acute coronary syndrome (ACS). METHODS: Two hundred and three patients, without previous cardiological follow-up and referred to the outpatient clinic after hospitalization for ACS between 9/1/2001 and 12/31/2002, were randomly allocated to a 12-week, 3-times-a-week CRP or standard cardiology follow-up only, after giving their informed consent. Baseline population characteristics were recorded and, in patients with more than 4 years' education, QL and depression were evaluated during hospitalization and at one-year follow-up by self-reported responses to the SF-36 QL survey and the Beck Depression Inventory (BDI) respectively. Analysis was by intention-to-treat. RESULTS: 31 patients randomized to CRP, mean age 56 years (31-80), 84% male (group A) and 95 patients, mean age 58 years (33-86), 83 % male, allocated to standard follow-up (group B), responded to the questionnaires. There were no significant differences between the two groups in any of the variables evaluated by the questionnaires during hospitalization. At one-year evaluation, the average BDI score was lower in the CRP group (8 vs. 11, p = 0.05). The prevalence of depressive symptoms (37.5 vs. 56.1%) or severe depression (3.1% vs. 12.2%) was also lower, in this group, although not significantly. SF-36 found significant differences in the evaluation of Vitality (average 62 points in A vs. 47 in B, p < 0.02) and General Health (57 points in A vs. 46 points in B, p < 0.02); there were no differences in the other parameters evaluated. When the physical and mental health components were analyzed, a significant difference favoring CRP was found in mental health (70.6 vs. 56.9, p = 0.02) but not in physical health (62.9 in A vs. 56.4 in B, NS). CONCLUSIONS: Despite the small sample size, significant differences favorable to CRP after ACS were found. After one-year follow-up, patients referred to CRP have a better BDI score; the Vitality and General Health parameters, as well as the mental health component evaluated by SF-36, are also significantly improved after CRP. PMID- 16041966 TI - Evaluation of glucose metabolism profile in coronary disease. Prospective study in hospitalized patients. AB - BACKGROUND: In 1995, it was estimated that 4% of the adult population had a diagnosis of diabetes mellitus. The trend is for growing prevalence, with a prediction of 300 million individuals with diabetes diagnosed in 2025. There seems to be a large number of asymptomatic individuals with undiagnosed disorders of glucose metabolism. These disorders, whether diabetes mellitus or conditions considered as prediabetic, including impaired glucose tolerance or impaired fasting glucose, represent an important risk factor for coronary disease and worsen the prognosis of established disease. The aim of this study was to evaluate the prevalence of glucose metabolism disorders in individuals admitted to hospital with coronary disease and to determine whether a significant number of these patients had previously undiagnosed diabetes or prediabetic conditions. METHODS: We conducted a prospective study of 44 consecutive patients admitted to the Cardiology Department with a diagnosis of coronary disease, whether previously established or established at the time of admission. We measured morning plasma glucose in all patients, after at least eight hours fasting, and all except those with a previous diagnosis of diabetes underwent an oral glucose tolerance test with 75 g of glucose before hospital discharge. For classification of glucometabolic state, we used the values defined by the World Health Organization (1999). RESULTS: The mean age of our cohort was 64.2+/-13.6 years. The most prevalent disorder was diabetes, with 19 patients (43.2%). Of these, 12 patients (27.3%) had a previous diagnosis of diabetes and 7 patients (15.9%) were unaware that they had the disease. We identified isolated impaired glucose tolerance in 11 patients (25%), isolated impaired fasting glucose in one patient (2.3%) and combined impaired glucose tolerance and impaired fasting glucose in another (2.3%). The remaining 12 patients (27.3%) revealed normal glycemia values. CONCLUSIONS: Glucose metabolism disorders including diabetes and impaired glucose tolerance have a high prevalence in coronary patients. This population includes a significant number of asymptomatic patients with undiagnosed diabetes or undetected prediabetic conditions. A systematic evaluation of the glucometabolic state of individuals with coronary disease during hospitalization, using an oral glucose tolerance test, may enable earlier identification of these disorders and implementation of measures to reduce their future impact. PMID- 16041968 TI - Left ventricular dysfunction after acute myocardial infarction--the impact of cardiovascular risk factors. AB - INTRODUCTION: Left ventricular (LV) systolic function is an important prognostic factor in coronary heart disease. Left ventricular ejection fraction (LVEF) should be assessed in all patients after acute myocardial infarction (AMI). Although reperfusion therapy has been found effective in the reduction of complications of AMI, LVEF impairment is a common consequence of an acute coronary event. The aim of this study was to estimate the incidence of LVEF depression after ST-elevation myocardial infarction (STEMI) and to evaluate the effect of previous cardiovascular risk factors on the risk of LV dysfunction. METHODS: One hundred and forty-seven consecutive patients with a first STEMI were included in this study. Most patients were male (70.7%) and mean age was 60.7 years. LVEF was assessed by echocardiography (using the single-plane area-length method and automatic border detection). LV systolic function was considered depressed when ejection fraction was less than 45 %. The chi-square test was used in the statistical analysis to compare proportions and a logistic regression model was fitted to assess the independent effect of each variable. RESULTS: Incidence of LV dysfunction was 55.8% in STEMI patients. No association was found between gender or age and LVEF impairment. The proportion of patients with diabetes was higher in the impaired LVEF group than in normal LVEF patients (44.7% vs. 31.7%, p = 0.12); the prevalence of smoking was also higher in patients with LV dysfunction (46.9% vs. 33.8%, p = 0.11). On the other hand, dyslipidemia was less common in patients with depressed LV function (35.4% vs. 56.9%, p = 0.01). Hypertension was not associated with impaired LVEF. After adjustment for ST-elevation location and number of vessels with critical stenosis, diabetes and smoking were associated with a significantly higher risk of LVEF impairment (diabetes: OR = 3.73, 95% CI 1.25-11.16; smoking: OR = 3.9, 95% CI 1.37-11.07) and dyslipidemia with a significantly lower risk of LV dysfunction (OR: 0.37, 95% CI 0.15-0.88). CONCLUSIONS: In STEMI patients, previous cardiovascular risk factors have a significant impact on the likelihood of LV dysfunction and hence could influence long-term prognosis. PMID- 16041967 TI - Ventricular tachycardia ablation in patients with coronary heart disease: beyond the reentry circuit. AB - INTRODUCTION: Patients with coronary heart disease and left ventricular dysfunction are at increased risk for the development of ventricular tachycardia (VT) related to areas of myocardial fibrosis. Although the mechanism and the circuit of this arrhythmia are well understood, little is known about the triggers that precipitate VT episodes. Purkinje fiber potentials may be responsible for idiopathic VT, and recent studies have related them to polymorphic VT and ventricular fibrillation. METHODS: Between January 2002 and December 2003, we performed ablation in 10 patients with coronary heart disease, left ventricular systolic dysfunction and VT refractory to pharmacological therapy. All patients had implantable cardioverter-defibrillators. Electroanatomical activation and voltage mapping (CARTO) and electrophysiological criteria (premature activation during VT, pace mapping, and presence of diastolic potentials) were used to define scar regions, slow conduction areas and the reentry circuit isthmuses. RESULTS: Spike potentials were recorded in the scars of three patients. These potentials were almost fused with the ventricular electrogram during sinus rhythm, and were more premature during VT, probably reflecting local activation of Purkinje fibers. During ablation, we were able to dissociate the spike from the ventricular electrogram, thus terminating the VT. In the cases with conduction recovery, ventricular; ectopic beats recurred, preceded by a spike and degenerating into short runs of VT. The ablation strategy was not modified since persistence of the VT required the isthmus. CONCLUSION: The results suggest that residual Purkinje fibers may be present in scar regions and that the activity of these fibers may trigger VT in pre-established circuits. PMID- 16041969 TI - Cholesterol pericarditis--relapsing pericardial effusion in a patient with rheumatoid arthritis. AB - Cholesterol pericarditis is an uncommon form of pericardial disease, of unknown pathophysiology, that is characterized by chronic relapsing, usually large, pericardial effusions that are distinctive due to a high level of cholesterol. Usually it is idiopathic, but it can be associated with various systemic diseases such as hypothyroidism, rheumatoid arthritis and tuberculosis, among others. Its clinical course is usually indolent and complications such as cardiac tamponade and chronic constrictive pericarditis are relatively rare. However, the need for surgery for complete treatment has been reported in at least 10 % of cases. When rheumatoid arthritis is the underlying cause, this outcome is more frequent among those with an acute episode of pericarditis during the course of the disease. We report the case of a 61-year-old female rheumatoid arthritis patient, who presented with heart failure due to a large pericardial effusion and was successfully treated by a surgical approach. PMID- 16041970 TI - Thermoregulation and aging. AB - Worldwide climate changes, bringing long-lasting higher and lower extremes of temperature, are leading to an increased mortality rate in elderly people due to cardiocirculatory dysfunction. The ability of young adults to make physiological adjustments when exposed to low or high temperatures decreases with age. The purpose of this literature review is to assess the dysfunctions occurring in the elderly as a result of their reduced responsiveness to thermal stress. Physiological responses to cold environment induce a reversible plasma water shift from the intravascular to the interstitial and intracellular level due to peripheral vasoconstriction. This phenomenon occurs on exposure to cold water or air and even during facial cooling by a cold wind. Above-normal temperatures induce a reduction in intravascular water due to sweating. Older people are less able to cope with the increased viscosity of blood and are thus more liable to suffer a cardiocirculatory accident. PMID- 16041971 TI - Studies of harm. Clinical applications. AB - When a clinician institutes a therapy, he/she has to consider, besides its efficacy, the harm a substance that is foreign to the body may cause. Nowadays, with the number of drugs available, more and more iatrogenic events are occurring, to the extent that the term 'iatroepidemic' has been coined to describe the phenomenon. Harm can be assessed by experimental studies, but this design raises ethical as well as financial issues, so the best evidence is gathered from observational data, mainly case-control and cohort studies. In previous articles we discussed the methodology to evaluate therapeutic benefit. In this article we aim to review designs for determining harm. PMID- 16041972 TI - Multiple left ventricular masses in a patient with asymptomatic ventricular arrhythmia and cutaneous follicular hamartoma. PMID- 16041973 TI - Three-dimensional reconstruction of mitral prosthesis flow. PMID- 16041974 TI - [Finally, is it worth the effort to be a member of the Swiss Veterinary Association?]. PMID- 16041975 TI - [Kick injuries of veterinarians during examination and treatment of horses: a retrospective study in Switzerland]. AB - The risk of injury to veterinarians treating horses is high, particularly when performing painful procedures. The present retrospective study investigated this risk, including the frequency and severity of kick injuries, by submitting questionnaires to 700 veterinary practitioners in Switzerland. Evaluation of 216 completed questionnaires revealed that the frequency of kick injuries was affected by the professional experience of the veterinarian, veterinarians being an owner of a horse, the season, the type of procedure, the percentage of equine patients in the practice population, the treatment location (stall, pasture) and chemical restraint of the patient. Severe injuries incurred by veterinarians in recent years were few in number and the result of a number of unfortunate circumstances, which included hurried procedures due to time restraints and unforeseen events that frightened the patient. The risk factors derived from the full statistical model are the percentage of equine patients in the practice population, the treatment location, the type of procedure and chemical restraint of the patient. Using this information, it should be feasible to reduce the risk of injuries. Sedation of horses undergoing painful procedures, choosing a safe treatment location and caution on the part of the veterinarian are advised to reduce the number of injuries. PMID- 16041976 TI - [Prevalence of gastric lesions in the pars proventricularis in finishing pigs at slaughter in Switzerland]. AB - A cross-sectional study was conducted at two large abattoirs in the winter 2003/04 to determine the prevalence of gastric lesions in Swiss finishing pigs. The stomachs of 1897 pigs from 107 slaughter groups were examined for macroscopic epithelial lesion of the pars proventricularis. Severe hyperceratosis, erosion, ulceration and constriction at the transition between oesophagus and stomach were described as gastric lesions. Lesions were observed in 36% of all animals, with slaughter group prevalence ranging from 0% to 94%. Ulceration or constriction was present in four percent of the examined pigs. Associations between gastric lesions, farm management data, and data obtained at ante and post mortem examinations conducted at the slaughterhouse were analysed. A significantly higher prevalence of gastric lesions was found in pigs originating from conventional housing systems compared to pigs raised on farms with 'animal friendly' housing systems providing straw bedding and outdoor exercise to pigs. In addition, significantly higher prevalence was observed in pigs originating from herds with clinical signs of tail biting and peritarsitis. PMID- 16041977 TI - [Suspected adverse drug reactions in 2004]. AB - We received 62 reports of suspected adverse events (SARs) for the year 2004. Their number and repartition according to affected animals and active substances were comparable with the previous year. The distributors or manufacturers submitted most of the declarations, but the proportion submitted by practicing veterinarians is slowly growing. 72% of the declarations dealt with adverse reactions in small animals (cats and dogs), followed by cattle and horses. Antiparasitics, anti-inflammatory drugs and immunologicals were the most frequently mentioned therapeutic classes, which are listed here according to the international ATCvet classification. A report from abroad about a fatality following injection of an antibiotic preparation for cattle prompted Swissmedic to review the security of this medication also sold on the Swiss market. It was decided to modify the package insert to warn about the danger of inadvertent self injection in humans, to reduce the risk of similar accidents in Switzerland. We hope that the pharmacovigilance system will enjoy an increasing awareness by the practicing veterinarians. PMID- 16041978 TI - [Extraction of the incisors in a dwarf rabbit]. AB - The dwarf rabbit often suffers from dental malocclusion, particularly on the level of the incisors. A too short maxillary jawbone in these brachycephalic animals is the cause. The treatment of first intention consists in reducing the overgrowth using a diamond bur, but like the problem repeats quickly, an alternative consists in extracting the 6 incisors. PMID- 16041979 TI - Residual stress in hydroxyapatite coating: nonlinear analysis and high-energy synchrotron measurements. AB - The thermal deposition of hydroxyapatite (HA) on titanium alloy substrate (Ti-6A1 4V) leads to a structure that has very good osseointegration properties. However, clinical failures have been occasionally reported at the interface between substrate and coating. Lifetime is the main parameter in such prostheses; therefore, in order to improve their quality, it is necessary to evaluate the level of stresses near the interface. The high-energy synchrotron radiation combines the advantages of a bulk analysis and reduced volume of the gauge. The objective of our study was to calculate the residual stress using a nonlinear finite-element model and to measure residual stress level near the interface, in the hydroxyapatite coating and in titanium alloy substrate with a nondestructive and high-resolution experiment. The high-energy synchrotron radiation of the BM16 beam-line at ESRF (Grenoble-France) was used with a resolution of down to 10 micrometers. The experimental measurements validate the results found by means of nonlinear finite-element analysis of the plasma spraying induced stress. PMID- 16041980 TI - Interactive simulation of needle insertion models. AB - A novel interactive virtual needle insertion simulation is presented. The simulation models are based on measured planar tissue deformations and needle insertion forces. Since the force-displacement relationship is only of interest along the needle shaft, a condensation technique is shown to reduce the computational complexity of linear simulation models significantly. As the needle penetrates or is withdrawn from the tissue model, the boundary conditions that determine the tissue and needle motion change. Boundary condition and local material coordinate changes are facilitated by fast low-rank matrix updates. A large-strain elastic needle model is coupled to the tissue models to account for needle deflection and bending during simulated insertion. A haptic environment, based on these novel interactive simulation techniques, allows users to manipulate a three-degree-of-freedom virtual needle as it penetrates virtual tissue models, while experiencing steering torques and lateral needle forces through a planar haptic interface. PMID- 16041981 TI - Simulation of lung function evolution after heart-lung transplantation using a numerical model. AB - A morphometry-based computational model for expiratory flow in humans was used to study the unusual configuration of the maximum expiratory flow-volume (MEFV) curve associated with alterations in lung function after heart-lung transplantation (HLT). The postoperative MEFV curve showed a peak, followed by a gently sloping plateau over the midvolume range, ending in a knee where the flow suddenly fell, instead of the usual observed uniform decrease in expiratory flow. We have tested several hypotheses about the relationship between the pattern of changes in the configuration of the MEFV curve and pathological changes in the airway mechanics through computer simulations. Principally, effects of lung denervation and airway obstruction, associated with the development of bronchiolitis obliterans in the lung periphery, have been investigated. The calculated curves are similar in appearance to the measured postoperative flow volume curves and confirm reliability of the earlier hypotheses. We conclude that the plateau-knee configuration of the MEFV curve can result from flow limitation in one of the first airway generations, that this flow limitation coupled with an increase in peripheral airway resistance results in plateau shortening, and that flows exceeding predicted values during the second part of expiration may be produced by lung denervation. Additionally our results demonstrate that airways larger than the transitional and respiratory bronchioles can be involved in pulmonary function deterioration observed in patients affected with obliterative bronchiolitis. Our findings indicate that the computational model, based on a symmetrical dichotomous branching structure of the bronchial tree, along with pathological data, can be employed to evaluate the effects of heterogeneous changes in the lung periphery. Index Terms-Airway mechanics, forced expiration, lung transplantation, mathematical modeling, maximal expiratory flow-volume curve. PMID- 16041982 TI - Detection of repolarization alternans with an implantable cardioverter defibrillator lead in a porcine model. AB - Mechanistic links have been suggested between repolarization alternans (RPA) and the onset of ventricular tachycardia (VT) and/or fibrillation. Endocardial detection of RPA may, therefore, be an important step in future device-based treatments of arrhythmias. Here, we investigate if RPA could be detected during acute ischemia using an implantable cardioverter defibrillator (ICD) lead (tip to distal coil) located in the right ventricular apex. In 18 pigs, the right coronary (n = 10) or left anterior descending coronary (n = 8) artery was occluded for 10 min using a balloon catheter, followed by reperfusion for 30 min, and re-occlusion for 30 min. RPA magnitude, computed using the modified moving average (MMA) method, showed a sharp increase in all 18 animals, from a mean baseline level of 1.9 +/- 1.3 mV to 3.0 +/- 1.3 mV during first occlusion (p < 0.001). RPA magnitude showed a prominent increase in 10 animals during re occlusion, from a mean baseline level of 1.7 +/- 1.0 mV to 3.3 +/- 1.5 mV (p < 0.001). The protocol was terminated during the first two stages of occlusion and reperfusion for the remaining 8 animals due to the occurrence of ventricular fibrillation (VF). These results confirm that RPA increases under ischemic conditions and that it is possible to detect and track RPA dynamics with an ICD lead that is positioned in a clinically realistic location. Such an approach may be useful in formulating improved arrhythmia detection and control algorithms. PMID- 16041983 TI - Application of higher order statistics techniques to EMG signals to characterize the motor unit action potential. AB - The electromyographic (EMG) signal provides information about the performance of muscles and nerves. At any instant, the shape of the muscle signal, motor unit action potential (MUAP), is constant unless there is movement of the position of the electrode or biochemical changes in the muscle due to changes in contraction level. The rate of neuron pulses, whose exact times of occurrence are random in nature, is related to the time duration and force of a muscle contraction. The EMG signal can be modeled as the output signal of a filtered impulse process where the neuron firing pulses are assumed to be the input of a system whose transfer function is the motor unit action potential. Representing the neuron pulses as a point process with random times of occurrence, the higher order statistics based system reconstruction algorithm can be applied to the EMG signal to characterize the motor unit action potential. In this paper, we report results from applying a cepstrum of bispectrum based system reconstruction algorithm to real wired-EMG (wEMG) and surface-EMG (sEMG) signals to estimate the appearance of MUAPs in the Rectus Femoris and Vastus Lateralis muscles while the muscles are at rest and in six other contraction positions. It is observed that the appearance of MUAPs estimated from any EMG (wEMG or sEMG) signal clearly shows evidence of motor unit recruitment and crosstalk, if any, due to activity in neighboring muscles. It is also found that the shape of MUAPs remains the same on loading. PMID- 16041984 TI - MEG forward problem formulation using equivalent surface current densities. AB - We present a formulation for the magnetoencephalography (MEG) forward problem with a layered head model. Traditionally the magnetic field is computed based on the electric potential on the interfaces between the layers. We propose to express the effect of the volumetric currents in terms of an equivalent surface current density on each interface, and obtain the magnetic field based on them. The boundary elements method is used to compute the equivalent current density and the magnetic field for a realistic head geometry. We present numerical results showing that the MEG forward problem is solved correctly with this formulation, and compare it with the performance of the traditional formulation. We conclude that the traditional formulation generally performs better, but still the new formulation is useful in certain situations. PMID- 16041985 TI - Time-frequency characterization of interdependencies in nonstationary signals: application to epileptic EEG. AB - For the past decades, numerous works have been dedicated to the development of signal processing methods aimed at measuring the degree of association between electroencephalographic (EEG) signals. This interdependency parameter, which may be defined in various ways, is often used to characterize a functional coupling between different brain structures or regions during either normal or pathological processes. In this paper, we focus on the time-frequency characterization of the interdependency between signals. Particularly, we propose a novel estimator of the linear relationship between nonstationary signals based on the cross correlation of narrow band filtered signals. This estimator is compared to a more classical estimator based on the coherence function. In a simulation framework, results show that it may exhibit better statistical performances (bias and variance or mean square error) when a priori knowledge about time delay between signals is available. On real data (intracerebral EEG signals), results show that this estimator may also enhance the readability of the time-frequency representation of relationship and, thus, can improve the interpretation of nonstationary interdependencies in EEG signals. Finally, we illustrate the importance of characterizing the relationship in both time and frequency domains by comparing with frequency-independent methods (linear and nonlinear). PMID- 16041986 TI - Monte Carlo simulation of light-tissue interaction: three-dimensional simulation for trans-illumination-based imaging of skin lesions. AB - Three-dimensional, voxel-based, and wavelength-dependent skin lesion models are developed and simulated using Monte Carlo techniques. The optical geometry of the Nevoscope with trans-illumination is used in the simulations for characterizing the lesion thickness. Based on the correlation analysis between the lesion thickness and the diffuse reflectance, optical wavelengths are selected for multispectral imaging of skin lesions using the Nevoscope. Tissue optical properties reported by various researchers are compiled together to form a voxel library. Tissue models used in the simulations are developed using the voxel library which offers flexibility in updating the optical properties and adding new media types into the models independent of the Monte Carlo simulation code. PMID- 16041987 TI - Ultrawideband microwave breast cancer detection: a detection-theoretic approach using the generalized likelihood ratio test. AB - Microwave imaging has been suggested as a promising modality for early-stage breast cancer detection. In this paper, we propose a statistical microwave imaging technique wherein a set of generalized likelihood ratio tests (GLRT) is applied to microwave backscatter data to determine the presence and location of strong scatterers such as malignant tumors in the breast. The GLRT is formulated assuming that the backscatter data is Gaussian distributed with known covariance matrix. We describe the method for estimating this covariance matrix offline and formulating a GLRT for several heterogeneous two-dimensional (2-D) numerical breast phantoms, several three-dimensional (3-D) experimental breast phantoms, and a 3-D numerical breast phantom with a realistic half-ellipsoid shape. Using the GLRT with the estimated covariance matrix and a threshold chosen to constrain the false discovery rate (FDR) of the image, we show the capability to detect and localize small (<0.6 cm) tumors in our numerical and experimental breast phantoms even when the dielectric contrast of the malignant-to-normal tissue is below 2:1. PMID- 16041988 TI - A programmable microsystem using system-on-chip for real-time biotelemetry. AB - A telemetry microsystem, including multiple sensors, integrated instrumentation and a wireless interface has been implemented. We have employed a methodology akin to that for System-on-Chip microelectronics to design an integrated circuit instrument containing several "intellectual property" blocks that will enable convenient reuse of modules in future projects. The present system was optimized for low-power and included mixed-signal sensor circuits, a programmable digital system, a feedback clock control loop and RF circuits integrated on a 5 mm x 5 mm silicon chip using a 0.6 microm, 3.3 V CMOS process. Undesirable signal coupling between circuit components has been investigated and current injection into sensitive instrumentation nodes was minimized by careful floor-planning. The chip, the sensors, a magnetic induction-based transmitter and two silver oxide cells were packaged into a 36 mm x 12 mm capsule format. A base station was built in order to retrieve the data from the microsystem in real-time. The base station was designed to be adaptive and timing tolerant since the microsystem design was simplified to reduce power consumption and size. The telemetry system was found to have a packet error rate of 10(-3) using an asynchronous simplex link. Trials in animal carcasses were carried out to show that the transmitter was as effective as a conventional RF device whilst consuming less power. PMID- 16041989 TI - A closed-loop electrical stimulation system for cardiac cell cultures. AB - An integrated electrical stimulation and recording system was designed for closed loop control and analysis of cardiac cultures on planar microelectrode arrays. Stimulated action potentials from HL-1 clonal myocyte cultures were digitized, stimulation artifacts were removed using nulling and filtering methods, and analysis was performed to determine stimulation efficacy in real time. Results of this analysis were used to determine future stimulation waveform parameters such as polarity, amplitude, pulse duration, and rate or pattern. Algorithms were designed utilizing real-time analysis and control to maintain a desired electrophysiological response of the culture, such as an arbitrary capture fraction value. This paper presents the hardware and software design of the stimulus pulse circuitry, artifact extraction, analysis, and control components of the system. Applications of this technology include the study of cardiac cell physiology, improving the speed and accuracy of traditional open-loop stimulation protocols, pharmacological screening, and improving the performance of biosensors based on sensing electrical activity in cardiac cultures. PMID- 16041990 TI - The development and test of a device for the reconstruction of 3-D position and orientation by means of a kinematic sensor assembly with rate gyroscopes and accelerometers. AB - In this paper, we propose a device for the Position and Orientation (P&O) reconstruction of human segmental locomotion tasks. It is based on three mono axial accelerometers and three angular velocity sensors, geometrically arranged to form two orthogonal terns. The device was bench tested using step-by-step motor-based equipment. The characteristics of the six channels under bench test conditions were: crosstalk absent, non linearity < +/- 0.1% fs, hysteresis < 0.1% fs, accuracy 0.3% fs, overall resolution better than 0.04 deg/s, 2 x g x 10(-4). The device was validated with the stereophotogrammetric body motion analyzer during the execution of three different locomotion tasks: stand-to-sit, sit-to stand, gait-initiation. Results obtained comparing the trajectories of the two methods showed that the errors were lower than 3 x 10(-2) m and 2 deg during a 4s of acquisition and lower than 6 x 10(-3) m and 0.2 deg during the effective duration of a locomotory task; showing that the wearable device hereby presented permits the 3-D reconstruction of the movement of the body segment to which it is affixed for time-limited clinical applications. PMID- 16041991 TI - Design and performance issues of RF coils utilized in ultra high field MRI: experimental and numerical evaluations. AB - In this paper, two TEM resonators were evaluated experimentally and numerically at 8 tesla (T) (340 MHz for 1H imaging). The coils were constructed to be 21.2-cm long (standard) and 11-cm long (a proposed less claustrophobic design). The experimental evaluation was done on a single cadaver using an ultra high field, 8 T, whole-body magnet. The numerical modeling was performed using an in-house finite difference time domain packagethat treats the coil and the load (anatomically detailed human head model) as a single system. The coils were tested with quadrature excitation at different coil alignment positions with respect to human head. For head imaging at 8 T, the overall numerical and experimental results demonstrated that when compared to the longer coil, the shorter coil provides superior signal-to-noise ratio, coil sensitivity, and excite field in the biological regions that lie within both of the coils' structures. A study of the RF (excite/receive fields) homogeneity showed variations in the performance of both coils that are mostly dependant on the region of interest and the position of coil with respect to the head. As such, depending on the application, the shorter coil could be effectively utilized. PMID- 16041992 TI - Communication scheduling to minimize thermal effects of implanted biosensor networks in homogeneous tissue. AB - A network of biosensors can be implanted in a human body for health monitoring, diagnostics, or as a prosthetic device. Biosensors can be organized into clusters where most of the communication takes place within the clusters, and long range transmissions to the base station are performed by the cluster leader to reduce the energy cost. In some applications, the tissues are sensitive to temperature increase and may be damaged by the heat resulting from normal operations and the recharging of sensor nodes. Our work is the first to consider rotating the cluster leadership to minimize the heating effects on human tissues. We explore the factors that lead to temperature increase, and the process for calculating the specific absorption rate (SAR) and temperature increase of implanted biosensors by using the finite-difference time-domain (FDTD) method. We improve performance by rotating the cluster leader based on the leadership history and the sensor locations. We propose a simplified scheme, temperature increase potential, to efficiently predict the temperature increase in tissues surrounding implanted sensors. Finally, a genetic algorithm is proposed to exploit the search for an optimal temperature increase sequence. PMID- 16041993 TI - Impedance characterization and modeling of electrodes for biomedical applications. AB - A low electrode-electrolyte impedance interface is critical in the design of electrodes for biomedical applications. To design low-impedance interfaces a complete understanding of the physical processes contributing to the impedance is required. In this work a model describing these physical processes is validated and extended to quantify the effect of organic coatings and incubation time. Electrochemical impedance spectroscopy has been used to electrically characterize the interface for various electrode materials: platinum, platinum black, and titanium nitride; and varying electrode sizes: 1 cm2, and 900 microm2. An equivalent circuit model comprising an interface capacitance, shunted by a charge transfer resistance, in series with the solution resistance has been fitted to the experimental results. Theoretical equations have been used to calculate the interface capacitance impedance and the solution resistance, yielding results that correspond well with the fitted parameter values, thereby confirming the validity of the equations. The effect of incubation time, and two organic cell adhesion promoting coatings, poly-L-lysine and laminin, on the interface impedance has been quantified using the model. This demonstrates the benefits of using this model in developing better understanding of the physical processes occurring at the interface in more complex, biomedically relevant situations. PMID- 16041994 TI - Band-tunable and multiplexed integrated circuits for simultaneous recording and stimulation with microelectrode arrays. AB - Two thin-film microelectrode arrays with integrated circuitry have been developed for extracellular neural recording in behaving animals. An eight-site probe for simultaneous neural recording and stimulation has been designed that includes on chip amplifiers that can be individually bypassed, allowing direct access to the iridium sites for electrical stimulation. The on-probe amplifiers have a gain of 38.9 dB, an upper-cutoff frequency of 9.9 kHz, and an input-referred noise of 9.2 microV rms integrated from 100 Hz to 10 kHz. The low-frequency cutoff of the amplifier is tunable to allow the recording of field potentials and minimize stimulus artifact. The amplifier consumes 68 microW from +/- 1.5 V supplies and occupies 0.177 mm2 in 3 microm features. In vivo recordings have shown that the preamplifiers can record single-unit activity 1 ms after the onset of stimulation on sites as close as 20 microm to the stimulating electrode. A second neural recording array has been developed which multiplexes 32 neural signals onto four output data leads. Providing gain on this array eliminates the need for bulky headmounted circuitry and reduces motion artifacts. The time-division multiplexing circuitry has crosstalk between consecutive channels of less than 6% at a sample rate of 20 kHz per channel. Amplified, time-division-multiplexed multichannel neural recording allows the large-scale recording of neuronal activity in freely behaving small animals with minimum number of interconnect leads. PMID- 16041995 TI - Statistical encoding model for a primary motor cortical brain-machine interface. AB - A number of studies of the motor system suggest that the majority of primary motor cortical neurons represent simple movement-related kinematic and dynamic quantities in their time-varying activity patterns. An example of such an encoding relationship is the cosine tuning of firing rate with respect to the direction of hand motion. We present a systematic development of statistical encoding models for movement-related motor neurons using multielectrode array recordings during a two-dimensional (2-D) continuous pursuit-tracking task. Our approach avoids massive averaging of responses by utilizing 2-D normalized occupancy plots, cascaded linear-nonlinear (LN) system models and a method for describing variability in discrete random systems. We found that the expected firing rate of most movement-related motor neurons is related to the kinematic values by a linear transformation, with a significant nonlinear distortion in about 1/3 of the neurons. The measured variability of the neural responses is markedly non-Poisson in many neurons and is well captured by a "normalized Gaussian" statistical model that is defined and introduced here. The statistical model is seamlessly integrated into a nearly-optimal recursive method for decoding movement from neural responses based on a Sequential Monte Carlo filter. PMID- 16041996 TI - Simulation analysis of conduction block in unmyelinated axons induced by high frequency biphasic electrical currents. AB - Nerve conduction block induced by high-frequency biphasic electrical currents is analyzed using a lumped circuit model of the unmyelinated axon based on Hodgkin Huxley equations. Axons of different diameters (5-20 microm) can not be blocked completely when the stimulation frequency is between 2 kHz and 4 kHz. However, when the stimulation frequency is above 4 kHz, all axons can be blocked. At high frequency a higher stimulation intensity is needed to block nerve conduction. The larger diameter axon has a lower threshold intensity for conduction block. The stimulation waveform in which the pulsewidth changes with frequency is more effective in blocking nerve conduction than the waveform in which the pulsewidth is fixed. The activation of potassium channels, rather than inactivation of sodium channels, is the possible mechanism underlying the nerve conduction block of the unmyelinated axon. This simulation study further increases our understanding of axonal conduction block induced by high-frequency biphasic currents, and can guide future animal experiments as well as optimize stimulation waveforms that might be used for electrical nerve block in clinical applications. PMID- 16041997 TI - Accessing ampli-tonotopic organization of rat auditory cortex by microstimulation of cochlear nucleus. AB - Auditory brainstem implants (ABI) that electrically stimulate the surface of cochlear nucleus have been clinically used for the rehabilitation of deaf patients with bilateral vestibular schwannomas. The change of pitch perception with an active electrode location is not as clear in ABIs as in cochlear implants, a factor which might play a role in poorer speech performance in ABIs. The objective of present work was to develop an animal ABI model that could provide physiological data for future ABI development and optimization. The experimental system included a penetrating microelectrode array for microstimulation of the cochlear nucleus and a surface microelectrode array for mapping evoked potentials over the auditory cortex. We first obtained tone-evoked cortical activation patterns, which represented a place code of the frequency and intensity of test tones, i.e., the ampli-tonotopic organization, and compared the patterns with those evoked by cochlear nuclear microstimulation. Our experimental results demonstrated that microstimulation of both the dorsal and ventral cochlear nucleus (DCN and VCN) could access the cortical ampli-tonotopic organization as acoustic stimuli did. We also found that the cortical dynamic range was wider for the DCN than VCN stimulation and for the low-frequency than for the high-frequency pathway. The present results have great implications for improved ABI performance. PMID- 16041998 TI - An automated approach for analyzing D-periods in collagen fibril images. AB - This paper considers an approach for analyzing fibrillar collagen structures based on fundamental concepts of pattern recognition. It focuses on the quantitative comparison between collagen structural data (electron-optical data) and chemical data. Theoretical models in the form of sequence-generated histograms are used as reference for extracting and analyzing the structural unit in images from collagen fibrils. In this respect, collagen provides a valuable model system for studying the chemical basis of ultrastructure, as well as detecting the alterations in collagen fibril structure produced by a disorder. Application examples are presented and the results are compared with biochemical studies. PMID- 16041999 TI - A high-throughput 3-D composite dielectrophoretic separator. AB - Dielectrophoresis has great potential to offer a range of diverse fields from bioprocessing to clinical medicine, but is hampered by low throughput rates to the micrometer scale of the electrodes required to generate highly nonuniform fields. Here we describe a novel approach to electrode construction, using a drilled laminated structure to form channels bearing electrodes 30 microm across and 150 microm apart. Since these electrodes appear along all sides of the drilled bore, the trapping efficiency is improved over conventional devices. We have developed and demonstrated a separator capable of sorting a 50:50 mixture of viable and nonviable yeast cells into an 86:14 mixture at 25 mLhr(-1). PMID- 16042000 TI - Processing of pulse oximeter data using discrete wavelet analysis. AB - A wavelet-based signal processing technique was employed to improve an implantable blood perfusion monitoring system. Data was acquired from both in vitro and in vivo sources: a perfusion model and the proximal jejunum of an adult pig. Results showed that wavelet analysis could isolate perfusion signals from raw, periodic, in vitro data as well as fast Fourier transform (FFT) methods. However, for the quasi-periodic in vivo data segments, wavelet analysis provided more consistent results than the FFT analysis for data segments of 50, 10, and 5 s in length. Wavelet analysis has thus been shown to require less data points for quasi-periodic data than FFT analysis making it a good choice for an indwelling perfusion monitor where power consumption and reaction time are paramount. PMID- 16042001 TI - Automatic grading of retinal vessel caliber. AB - New clinical studies suggest that narrowing of the retinal blood vessels may be an early indicator of cardiovascular diseases. One measure to quantify the severity of retinal arteriolar narrowing is the arteriolar-to-venular diameter ratio (AVR). The manual computation of AVR is a tedious process involving repeated measurements of the diameters of all arterioles and venules in the retinal images by human graders. Consistency and reproducibility are concerns. To facilitate large-scale clinical use in the general population, it is essential to have a precise, efficient and automatic system to compute this AVR. This paper describes a new approach to obtain AVR. The starting points of vessels are detected using a matched Gaussian filter. The detected vessels are traced with the help of a combined Kalman filter and Gaussian filter. A modified Gaussian model that takes into account the central light reflection of arterioles is proposed to describe the vessel profile. The width of a vessel is obtained by data fitting. Experimental results indicate a 97.1% success rate in the identification of vessel starting points, and a 99.2% success rate in the tracking of retinal vessels. The accuracy of the AVR computation is well within the acceptable range of deviation among the human graders, with a mean relative AVR error of 4.4%. The system has interested clinical research groups worldwide and will be tested in clinical studies. PMID- 16042002 TI - An autocorrelation-based time domain analysis technique for monitoring perfusion and oxygenation in transplanted organs. AB - In designing an implantable sensor for perfusion monitoring of transplant organs the ability of the sensor to gather perfusion information with limited power consumption and in near real time is paramount. The following work was performed to provide a processing method that is able to predict perfusion and oxygenation change within the blood flowing through a transplanted organ. For this application, an autocorrelation-based algorithm was used to reduce the acquisition time required for fast Fourier transform (FFT) analysis while retaining the accuracy inherent to FFT analysis. In order to provide data proving that the developed method is able to predict perfusion as accurately as FFT two experiments were developed isolating both periodic and quasi-periodic cardiac frequencies. It was shown that the autocorrelation-based method was able to perform comparably with FFT (limited to a sampling frequency of 300 Hz) and maintain accuracy down to acquisition times as low as 4 s in length. PMID- 16042003 TI - Speaker normalization for chinese vowel recognition in cochlear implants. AB - Because of the limited spectra-temporal resolution associated with cochlear implants, implant patients often have greater difficulty with multitalker speech recognition. The present study investigated whether multitalker speech recognition can be improved by applying speaker normalization techniques to cochlear implant speech processing. Multitalker Chinese vowel recognition was tested with normal-hearing Chinese-speaking subjects listening to a 4-channel cochlear implant simulation, with and without speaker normalization. For each subject, speaker normalization was referenced to the speaker that produced the best recognition performance under conditions without speaker normalization. To match the remaining speakers to this "optimal" output pattern, the overall frequency range of the analysis filter bank was adjusted for each speaker according to the ratio of the mean third formant frequency values between the specific speaker and the reference speaker. Results showed that speaker normalization provided a small but significant improvement in subjects' overall recognition performance. After speaker normalization, subjects' patterns of recognition performance across speakers changed, demonstrating the potential for speaker-dependent effects with the proposed normalization technique. PMID- 16042004 TI - Induced current bio-impedance technique for monitoring cryosurgery procedure in a two-dimensional head model using generalized coordinate systems. AB - In the noninvasive bio-impedance technique, small amplitude currents are applied to the body and the developing potentials on its surface are measured. This noninvasive technique is used to monitor physiological and pathological processes, which alter the values or the spatial distribution of the electrical impedance inside the human body. A possible application of the bio-impedance technique is monitoring brain cryosurgery procedure--a surgical technique that employs freezing to destroy undesirable tissues. A numerical solver was developed to evaluate the ability of an induced-current bio-impedance system to monitor the growth of the frozen tissue inside the head in simulation. The forward-problem bio-impedance solver, which is based on the finite volume method in generalized two-dimensional (2-D) coordinate systems, was validated by a comparison to a known analytical solution for body-fitted and Cartesian meshing grids. The sensitivity of the developed surface potential to the ice-ball area was examined using a 2-D head model geometry, and was found to range between 0.8 x 10(-2) and 1.68 x 10(-2) (relative potential difference/mm2), depending on the relative positioning of the excitation coil and the head. The maximal sensitivity was achieved when the coil was located at the geometrical center of the model. PMID- 16042005 TI - Design and initial evaluation of an implantable sonomicrometer and CW Doppler flowmeter for simultaneous recordings with a multichannel telemetry system. AB - We have developed a sonomicrometer and continuous wavelength (CW) Doppler flowmeter for a multichannel telemetry system. These developments will enable us to measure ventricular dimension and coronary artery blood velocity, which are valuable parameters to characterize sudden cardiac death (SCD) in ambulatory animal models of ventricular arrhythmias. The design goals for the sensors were accuracy, low power consumption, small size and compatibility with each other. The circuits were designed successfully and tested simultaneously in vivo. The CW Doppler flowmeter draws 9 mA and the sonomicrometer draws 28 mA on a 5-V supply. The ability to measure heart dimension and blood velocity will add significantly to our understanding of the sequence of events leading up to spontaneous sudden cardiac death. PMID- 16042006 TI - Application of RNA interference to root-knot nematode genes encoding esophageal gland proteins. AB - Plant parasitic nematodes have been, so far, refractory to transformation or mutagenesis. The functional analysis of nematode genes relies on the development of reverse genetic tools adapted to these obligate parasites. Here, we describe the application of RNA interference (RNAi) to the root-knot nematode Meloidogyne incognita for the knock-down of two genes expressed in the subventral esophageal glands of the nematode and potentially involved in parasitism, the calreticulin (Mi-crt) and the polygalacturonase (Mi-pg-1) genes. Incubation in 1% resorcinol for 4 h induced double-stranded RNA uptake through the alimentary track of the nematodes and led to up to 92% depletion of Mi-crt transcripts. Timecourse analysis of the silencing showed different temporal patterns for Mi-crt and Mi-pg 1. The silencing of Mi-crt was optimal 20 h after soaking, whereas the silencing of Mi-pg-1 was optimal 44 h after soaking. For the two genes, the silencing effect was highly time-limited, since no transcript depletion was detectable 68 h after soaking. PMID- 16042007 TI - Functional analysis of pathogenicity proteins of the potato cyst nematode Globodera rostochiensis using RNAi. AB - RNA interference (RNAi) has been used widely as a tool for examining gene function and a method that allows its use with plant-parasitic nematodes recently has been described. Here, we use a modified method to analyze the function of secreted beta-1,4, endoglucanases of the potato cyst nematode Globodera rostochiensis, the first in vivo functional analysis of a pathogenicity protein of a plant-parasitic nematode. Knockout of the beta-1,4, endoglucanases reduced the ability of the nematodes to invade roots. We also use RNAi to show that gr ams-1, a secreted protein of the main sense organs (the amphids), is essential for host location. PMID- 16042008 TI - The Pseudomonas syringae avrRpt2 gene contributes to virulence on tomato. AB - In order to cause disease on plants, gram-negative phytopathogenic bacteria introduce numerous virulence factors into the host cell in order to render host tissue more hospitable for pathogen proliferation. The mode of action of such bacterial virulence factors and their interaction with host defense pathways remain poorly understood. avrRpt2, a gene from Pseudomonas syringae pv. tomato JL1065, has been shown to promote the virulence of heterologous P. syringae strains on Arabidopsis thaliana. However, the contribution of avrRpt2 to the virulence of JL1065 has not been examined previously. We show that a mutant derivative of JL1065 that carries a disruption in avrRpt2 is impaired in its ability to cause disease on tomato (Lycopersicon esculentum), indicating that avrRpt2 also acts as a virulence gene in its native strain on a natural host. The virulence activity of avrRpt2 was detectable on tomato lines that are defective in either ethylene perception or the accumulation of salicylic acid, but could not be detected on a tomato mutant insensitive to jasmonic acid. The enhanced virulence conferred by the expression of avrRpt2 in JL1065 was not associated with the suppression of several defense-related genes induced during the infection of tomato. PMID- 16042009 TI - Analysis of promoters recognized by HrpL, an alternative sigma-factor protein from Pantoea agglomerans pv. gypsophilae. AB - HrpL, an alternative sigma factor, activates the transcription of the Hrp regulon by its binding to a common "hrp box" promoter. Based on computational techniques, the hrp box previously was defined as a consensus bipartite cis element, 5' GGAACC-N(15-16)-CCACNNA-3'. The present report combines a quantitative in vivo assay for measuring Hrp promoter activity with site-specific mutagenesis to analyze the effect of consensus and nonconsensus nucleotides on promoter activity. The analysis was carried out with Hop effectors of the tumorigenic bacterium Pantoea agglomerans pv. gypsophilae, in which HrpL is indispensable for gall formation. Mutational analysis indicates that the hrp box consensus can be divided into crucial and noncrucial nucleotides. The first 5 nucleotides (nt) of the--35 consensus motif (GGAAC) and the 3 nt of the--10 motif (ACNNA) are crucial, whereas other consensus and adjacent nonconsensus nucleotides exert a significant effect on the promoter's strength. With spacing of 13 or 17 nt between the two motifs, significant activity was still retained. Gel shift assays indicated that deletion of GG from the--35 consensus motif eliminated HrpL binding, whereas mutations in the--10 consensus motif or modification of the spacing, which eliminates promoter activity, did not elicit any effect. The degeneracy in Hrp promoters of four hrp and type III effector genes of P agglomerans pv. gypsophilae indicated significant differences in promoter activity, whereas increasing the promoter strength of the Hop effector, HsvG, resulted in overexpression of gall formation. PMID- 16042010 TI - Growth deficiency of a Xanthomonas oryzae pv. oryzae fur mutant in rice leaves is rescued by ascorbic acid supplementation. AB - Xanthomonas oryzae pv. oryzae causes bacterial leaf blight, a serious disease of rice. A mutation was isolated in the ferric uptake regulator (fur) gene of X. oryzae pv. oryzae and it was shown to result in the production of siderophores in a constitutive manner. The fur mutant is hypersensitive to the metallo-antibiotic streptonigrin, a phenotype that is indicative of intracellular free-iron overload, and also exhibits a slow growth phenotype on rich medium. The fur mutant is virulence deficient, hypersensitive to hydrogen peroxide, and exhibits reduced catalase activity. Exogenous supplementation with ascorbic acid (an antioxidant) rescues the growth deficiency of the fur mutant in rice leaves. The virulence deficiency of the X. oryzae pv. oryzae fur mutant is proposed to be due, at least in part, to an impaired ability to cope with the oxidative stress conditions that are encountered during infection. PMID- 16042011 TI - The zinc uptake regulator Zur is essential for the full virulence of Xanthomonas campestris pv. campestris. AB - Zur is a regulator of the high-affinity zinc uptake system in many bacteria. In Xanthomonas campestris pv. campestris 8004, a putative protein encoded by the open reading frame designated as XC1430 shows 42% amino acid similarity with the Zur of Escherichia coli. An XC1430-disrupted mutant 1430nk was constructed by homologous suicide plasmid integration. 1430nk failed to grow in rich medium supplemented with Zn2+ at a concentration of 400 microM and in nonrich medium supplemented with Zn2+ at a concentration of 110 microM, whereas the wild-type strain grew well in the same conditions. In rich medium with 400 microM Zn2+, 1430nk accumulated significantly more Zn2+ than the wild-type strain. 1430nk showed a reduction in virulence on the host plant Chinese radish (Raphanus sativus L. var. radiculus Pers.) and produced less extracellular polysaccharide (EPS) than did the wild-type strain in the absence of added zinc. These results revealed that XC1430 is a functional member of the Zur regulator family that controls zinc homeostasis, EPS production, and virulence in X. campestris pv. campestris. PMID- 16042012 TI - Global patterns of gene regulation associated with the development of ectomycorrhiza between birch (Betula pendula Roth.) and Paxillus involutus (Batsch) Fr. AB - The formation of ectomycorrhizal (ECM) root tissue is characterized by distinct morphological and developmental stages, such as preinfection and adhesion, mantle, and Hartig net formation. The global pattern of gene expression during these stages in the birch (Betula pendula)-Paxillus involutus ECM association was analyzed using cDNA microarrays. In comparison with nonsymbiotic conditions, 251 fungal (from a total of 1,075) and 138 plant (1,074 in total) genes were found to be differentially regulated during the ECM development. For instance, during mantle and Hartig net development, there were several plant genes upregulated that are normally involved in defense responses during pathogenic fungal challenges. These responses were, at later stages of ECM development, found to be repressed. Other birch genes that showed differential regulation involved several homologs that usually are implicated in water permeability (aquaporins) and water stress tolerance (dehydrins). Among fungal genes differentially upregulated during stages of mantle and Hartig net formation were homologs putatively involved in mitochondrial respiration. In fully developed ECM tissue, there was an upregulation of fungal genes related to protein synthesis and the cytoskeleton assembly machinery. This study highlights complex molecular interactions between two symbionts during the development of an ECM association. PMID- 16042013 TI - Characterization of the Xanthomonas campestris pv. campestris lipopolysaccharide substructures essential for elicitation of an oxidative burst in tobacco cells. AB - The lipopolysaccharides (LPS) of gram-negative bacteria are essential for perception of pathogens by animals and plants. To identify the LPS substructure or substructures recognized by plants, we isolated water-phase (w)LPS from different Xanthomonas campestris pv. campestris mutants and analyzed their sugar content and ability to elicit an oxidative burst in tobacco cell cultures. The different wLPS species are characterized by lacking repetitive subunits of the O antigen, the complete O-antigen, or even most of the core region. Because loss of lipid A would be lethal to bacteria, pure lipid A was obtained from X. campestris pv. campestris wild-type wLPS by chemical hydrolysis. The elicitation experiments with tobacco cell cultures revealed that LPS detection is dependent on the bioavailability of the amphiphilic wLPS, which can form micelles in an aqueous environment. By adding deoxycholate to prevent micelle formation, all of the tested wLPS species showed elicitation capability, whereas the lipid A alone was not able to trigger an oxidative burst or calcium transients in tobacco cell cultures. These results suggest that the LPS substructure recognized by tobacco cells is localized in the inner core region of the LPS, consisting of glucose, galacturonic acid, and 3-deoxy-d-manno-oct-2-ulosonic acids. Although lipid A alone seems to be insufficient to induce an oxidative burst in tobacco cell cultures, it cannot be ruled out that lipid A or the glucosamine backbone may be important in combination with the inner core structures. PMID- 16042014 TI - Quorum sensing regulates exopolysaccharide production, motility, and virulence in Pseudomonas syringae. AB - The N-acyl homoserine lactone (AHL)-mediated quorum-sensing system in the phytopathogen Pseudomonas syringae pv. syringae requires the AHL synthase AhlI and the regulator AhlR, and is additionally subject to regulation by AefR. The contribution of quorum sensing to the expression of a variety of traits expected to be involved in epiphytic fitness and virulence of P syringae were examined. Both an aefR- mutant and an ahlI- ahlR- double mutant, deficient in AHL production, were significantly impaired in alginate production and had an increased susceptibility to hydrogen peroxide compared with the wild-type strain. These mutants were hypermotile in culture, invaded leaves more rapidly, and caused an increased incidence of brown spot lesions on bean leaves after a 48-h moist incubation. Interestingly, an aefR- mutant was both the most motile and virulent. Like the wild-type strain, the AHL-deficient mutant strains incited water-soaked lesions on bean pods. However, lesions caused by an ahlI- ahlR- double mutant were larger, whereas those incited by an aefR- mutant were smaller. In contrast, tissue maceration of pods, which occurs at a later stage of infection, was completely abolished in the AHL-deficient mutants. Both the incidence of disease and in planta growth of P syringae pv. tabaci were greatly reduced in transgenic tobacco plants that produced AHL compared with wild-type plants. These results demonstrate that quorum sensing in E syringae regulates traits that contribute to epiphytic fitness as well as to distinct stages of disease development during plant infection. PMID- 16042015 TI - Role of trehalose transport and utilization in Sinorhizobium meliloti--alfalfa interactions. AB - Genes thuA and thuB in Sinorhizobium meliloti Rm1021 code for a major pathway for trehalose catabolism and are induced by trehalose but not by related structurally similar disaccharides like sucrose or maltose. S. meliloti strains mutated in either of these two genes were severely impaired in their ability to grow on trehalose as the sole source of carbon. ThuA and ThuB show no homology to any known enzymes in trehalose utilization. ThuA has similarity to proteins of unknown function in Mesorhizobium loti, Agrobacterium tumefaciens, and Brucella melitensis, and ThuB possesses homology to dehydrogenases containing the consensus motif AGKHVXCEKP. thuAB genes are expressed in bacteria growing on the root surface and in the infection threads but not in the symbiotic zone of the nodules. Even though thuA and thuB mutants were impaired in competitive colonization of Medicago sativa roots, these strains were more competitive than the wild-type Rml021 in infecting alfalfa roots and forming nitrogen-fixing nodules. Possible reasons for their increased competitiveness are discussed. PMID- 16042016 TI - Live-cell imaging of rhabdovirus-induced morphological changes in plant nuclear membranes. AB - Potato yellow dwarf virus (PYDV) and Sonchus yellow net virus (SYNV) belong to the genus Nucleorhabdovirus. These viruses replicate in nuclei of infected cells and mature virions accumulate in the perinuclear space after budding through the inner nuclear membrane. Infection of transgenic Nicotiana benthamiana 16c plants (which constitutively express green fluorescent protein (GFP) targeted to endomembranes) with PYDV or SYNV resulted in virus-specific patterns of accumulation of both GFP and membranes within nuclei. Using immunolocalization and a lipophilic fluorescent dye, we show that the sites of the relocalized membranes were coincident with foci of accumulation of the SYNV nucleocapsid protein. In contrast to the effects of PYDV and SYNV, inoculation of 16c plants with plus-strand RNA viruses did not result in accumulation of intranuclear GFP. Instead, such infections resulted in accumulation of GFP around nuclei, in a manner consistent with proliferation of the endoplasmic reticulum. We propose that the relocalization of GFP in 16c plants can be used to study sites of rhabdovirus accumulation in live cells. This study is the first to use live-cell imaging to characterize the effects of rhabdoviruses on plant nuclear membranes. PMID- 16042017 TI - Visualization of interactions between a pathogenic and a beneficial Fusarium strain during biocontrol of tomato foot and root rot. AB - The soilborne fungus Fusarium oxysporum f. sp. radicis-lycopersici causes tomato foot and root rot (TFRR), which can be controlled by the addition of the nonpathogenic fungus F. oxysporum Fo47 to the soil. To improve our understanding of the interactions between the two Fusarium strains on tomato roots during biocontrol, the fungi were labeled using different autofluorescent proteins as markers and subsequently visualized using confocal laser scanning microscopy. The results were as follows. i) An at least 50-fold excess of Fo47over F. oxysporum f. sp. radicis-lycopersici was required to obtain control of TFRR. ii) When seedlings were planted in sand infested with spores of a single fungus, Fo47 hyphae attached to the root earlier than those of F. oxysporum f. sp. radicis lycopersici. iii) Subsequent root colonization by F. oxysporum f. sp. radicis lycopersici was faster and to a larger extent than that by Fo47. iv) Under disease-controlling conditions, colonization of tomato roots by the pathogenic fungus was significantly reduced. v) When the inoculum concentration of Fo47 was increased, root colonization by the pathogen was arrested at the stage of initial attachment to the root. vi) The percentage of spores of Fo47 that germinates in tomato root exudate in vitro is higher than that of the pathogen F. oxysporum f. sp. radicis-lycopersici. Based on these results, the mechanisms by which Fo47 controls TFRR are discussed in terms of i) rate of spore germination and competition for nutrients before the two fungi reach the rhizoplane; ii) competition for initial sites of attachment, intercellular junctions, and nutrients on the tomato root surface; and iii) inducing systemic resistance. PMID- 16042018 TI - The late blight resistance locus Rpi-bib3 from Solanum bulbocastanum belongs to a major late blight R gene cluster on chromosome 4 of potato. AB - Late blight, caused by Phytophthora infestans, is one of the most devastating diseases in cultivated potato. Breeding of new potato cultivars with high levels of resistance to P. infestans is considered the most durable strategy for future potato cultivation. In this study, we report the identification of a new late blight resistance (R) locus from the wild potato species Solanum bulbocastanum. Using several different approaches, a high-resolution genetic map of the new locus was generated, delimiting Rpi-blb3 to a 0.93 cM interval on chromosome 4. One amplification fragment length polymorphism marker was identified that cosegregated in 1,396 progeny plants of an intraspecific mapping population with Rpi-blb3. For comparative genomics purposes, markers linked to Rpi-blb3 were tested in mapping populations used to map the three other late-blight R loci Rpi abpt, R2, and R2-like also to chromosome 4. Marker order and allelic conservation suggest that Rpi-blb3, Rpi-abpt, R2, and R2-like reside in the same R gene cluster on chromosome 4 and likely belong to the same gene family. Our findings provide novel insights in the evolution of R gene clusters conferring late-blight resistance in Solanum spp. PMID- 16042019 TI - Profiling of wheat class III peroxidase genes derived from powdery mildew attacked epidermis reveals distinct sequence-associated expression patterns. AB - A cDNA library was constructed from leaf epidermis of diploid wheat (Triticum monococcum) infected with the powdery mildew fungus (Blumeria graminis f. sp. tritici) and was screened for genes encoding peroxidases. From 2,500 expressed sequence tags (ESTs), 36 cDNAs representing 10 peroxidase genes (designated TmPRX1 to TmPRX10) were isolated and further characterized. Alignment of the deduced amino acid sequences and phylogenetic clustering with peroxidases from other plant species demonstrated that these peroxidases fall into four distinct groups. Differential expression and tissue-specific localization among the members were observed during the B. graminis f. sp. tritici attack using Northern blots and reverse-transcriptase polymerase chain reaction analyses. Consistent with its abundance in the EST collection, TmPRX1 expression showed the highest induction during pathogen attack and fluctuated in response to the fungal parasitic stages. TmPRX1 to TmPRX6 were expressed predominantly in mesophyll cells, whereas TmPRX7 to TmPRX10, which feature a putative C-terminal propeptide, were detectable mainly in epidermal cells. Using TmPRX8 as a representative, we demonstrated that its C-terminal propeptide was sufficient to target a green fluorescent protein fusion protein to the vacuoles in onion cells. Finally, differential expression profiles of the TmPRXs after abiotic stresses and signal molecule treatments were used to dissect the potential role of these peroxidases in multiple stress and defense pathways. PMID- 16042020 TI - GuaB activity is required in Rhizobium tropici during the early stages of nodulation of determinate nodules but is dispensable for the Sinorhizobium meliloti-alfalfa symbiotic interaction. AB - The guaB mutant strain Rhizobium tropici CIAT8999-10T is defective in symbiosis with common bean, forming nodules that lack rhizobial content. In order to investigate the timing of the guaB requirement during the nodule formation on the host common bean by the strain CIAT899-10.T, we constructed gene fusions in which the guaB gene is expressed under the control of the symbiotic promoters nodA, bacA, and nifH. Our data indicated that the guaB is required from the early stages of nodulation because full recovery of the wild-type phenotype was accomplished by the nodA-guaB fusion. In addition, we have constructed a guaB mutant derived from Sinorhizobium meliloti 1021, and shown that, unlike R. tropici, the guaB S. meliloti mutant is auxotrophic for guanine and induces wild type nodules on alfalfa and Medicago truncatula. The guaB R. tropici mutant also is defective in its symbiosis with Macroptilium atropurpureum and Vigna unguiculata but normal with Leucaena leucocephala. These results show that the requirement of the rhizobial guaB for symbiosis is found to be associated with host plants that form determinate type of nodules. PMID- 16042021 TI - George Combe--phrenologist, philosopher, psychologist (1788-1858). PMID- 16042022 TI - When is category specific in Alzheimer's disease? AB - Mixed findings have emerged concerning whether category-specific disorders occur in Alzheimer's disease. Factors that may contribute to these inconsistencies include: ceiling effects/skewed distributions for control data in some studies; differences in the severity of cognitive deficit in patients; and differences in the type of analysis (in particular, if and how controls are used to analyse single case data). We examined picture naming in Alzheimer's patients and matched elderly healthy normal controls in three experiments. These experiments used stimuli that did and did not produce ceiling effects/skewed data in controls. In Experiment 1, we examined for category effects in individual DAT patients using commonly used analyses for single cases (chi2 and z-scores). The different techniques produced quite different outcomes. In Experiment 2a, we used the same techniques on a different group of patients with similar outcomes. Finally, in Experiment 2b, we examined the same patients but (a) used stimuli that did not produce ceiling effects/skewed distributions in healthy controls, and (b) used statistical methods that did not treat the control sample as a population. We found that ceiling effects in controls may markedly inflate the incidence of dissociations in which living things are differentially impaired and seriously underestimate dissociations in the opposite direction. In addition, methods that treat the control sample as a population led to inflation in the overall number of dissociations detected. These findings have implications for the reliability of category effects previously reported both in Alzheimer patients and in other pathologies. In particular, they suggest that the greater proportion of living than nonliving deficits reported in the literature may be an artifact of the methods used. PMID- 16042023 TI - Information transmission defect identified and localized in language learning impaired children by means of electrophysiology. AB - Children with language processing deficits have various learning impairments and poor scholastic performance. In 3-10% of all children a specific language processing deficit can be identified by the Sound Connecting Sub-Test of the Illinois Test of Psycholinguistic-Abilities (SC-ITPA). These children among which we drew our index group (AS-Group) suffer from the disability to recognize isolated sounds as parts of words. Following linguistic terminology this is known as an auditory sequential sound processing deficit (ASSPD) Eighteen children (AS Group) and 21 controls (C-Group) were subjected to mapped P300 evoked potential analyses of cortical response to acoustic stimulation in the oddball paradigm. The data presented here show that there exists significant relation between the P300 amplitude reduction and ASSPD. The P300 amplitude decrease measured in the AS-Group is due to a reduced information transmission in accordance with Johnson's Triarchic Model of the P300 Amplitude. The cerebral structures involved in poor language processing are localized at the left temporo-parietal cortex. This supports the hypothesis that the underlying neuronal defect of ASSPD is localized in the language center and not in the auditory pathway. The P300 amplitude may serve as electrophysiological tool to identify ASSPD and to quantify the degree of improvement in the course of specific therapy. PMID- 16042024 TI - A visual disorder producing highly selective deletion of recurring letters. AB - A case of unusually selective deletion of recurring letters is described. The disorder was primarily visual, spelling being unaffected. There was no evidence of neglect. Reading was fluent and rapid (not letter-by-letter), though words often appeared as a series of resolving fragments. Selective deletions were entirely confined to adjacent letters, excluding explanation in terms of repetition blindness (Kaniwisher, 1991). Mixing case and font, or increasing inter-letter distance, did not alter rate of recurring letter deletion, provided the task required some form of perceptual grouping. The final 2 experiments establish the pre-lexical nature of the deficit; they show deletion of recurring symbols, for example, when copying Navon-style global forms constructed from same or different Greek letters. This visual deficit is explained using the neural network model developed by Rolls and Deco (2002). PMID- 16042025 TI - Effects of memory consolidation on human hippocampal activity during retrieval. AB - Day-to-day memories undergo transformation from short-term to long-term storage, a process called memory consolidation. Animal studies showed that memory consolidation requires protein synthesis and the growth of new hippocampal synapses within 24 h. To test for effects of memory consolidation in the human, we examined brain activation during the retrieval of information at 10 min and at 24 h following learning using functional magnetic resonance imaging (fMRI), an indirect measure of synaptic activity. Learning instructions were adjusted to yield a comparable retrieval quantity and retrieval quality at 10 min and 24 h after learning. The left hippocampal formation exhibited enhanced activity during the retrieval at the 24 h lag compared to the retrieval at the 10 min lag. Moreover, the activity in the left anterior hippocampal formation showed stronger correlations with retrieval quantity and retrieval quality at the 24 h lag than at the 10 min lag. This suggests that the relation between left anterior hippocampal activity and retrieval success became closer as consolidation progressed. These fMRI results in the human hippocampal formation may correspond to the neurobiological results in the animal hippocampal formation of a strengthening and growth of synaptic connections within 24 h. PMID- 16042026 TI - Unilateral visual cueing and asymmetric line geometry share a common attentional origin in the modulation of pseudoneglect. AB - Numerous factors influence the leftward bias (pseudoneglect) in perceived line midpoint of normal subjects in line bisection tasks. Cues are a potent factor; left and right cues promote shifts in perceived midpoint to the left and right, respectively. Trapezoidal lines have recently been shown to influence perceived line midpoint, displacing it toward the larger side. The present experiments test the hypothesis that the effect of line geometry, like that of unilateral cues, results from an exogenous recruitment of spatial attention. Normal right-handed subjects (N = 60) participated in two experiments employing a tachistoscopic forced-choice line bisection task. Experiment 1 crossed the effect of cue position and cue contrast, and confirmed that a significant interaction could be obtained. Experiment 2 crossed the effect of line geometry and cue position, revealing that line geometry and cue location both significantly influence perceived line midpoint, and produce a significant interaction. According to Additive Factors Logic the finding that spatial cueing interacts with line geometry suggests that both types of stimuli modulate spatial attention at a common site of processing, supporting the conclusion that the effect of line geometry itself derives from asymmetric cueing. An explanation for the interaction is offered that is based on the existence of a hypothesized compressive nonlinearity that maps attentional bias to perceptual error. PMID- 16042027 TI - The "zoom lens" of focal attention in visual search: changes in aging and Alzheimer's disease. AB - Visual search for a target in an array of distractors relies upon flexible shifts between global and local modes of attentional processing. Visual search is slowed in patients with Alzheimer's disease (AD), in part due to an increase in the number and duration of eye fixations made en route to a target (Rosler et al., 2000). This phenomenon may represent a compensatory adaptation to a narrowing of the zone of focal attention, necessitating more shifts of gaze in order to attend to the global workspace. Eye fixations were analyzed in two regions of interest (ROIs, central fixation and peripheral target locations) in 9 patients with mild AD, 9 cognitively intact age-matched control subjects, and 9 young controls, while they searched for a target object in a radial array that contained from 1 to 6 stimuli. Contrasted with young subjects, the search strategy of older controls and, to a greater extent, AD patients showed an increase in the average number and duration of peripheral fixations. Reduced efficiency of visual search in AD may be contributed to by reduced ability to dynamically adjust the attentional zoom, coupled with the inability to disengage attention from peripheral targets. PMID- 16042028 TI - Referential communication in Alzheimer's type dementia. AB - This paper investigates factors that underlie reduced informative content and lack of reference in the discourse of patients with Alzheimer's Type Dementia (DAT). Patients with DAT, fluent aphasics and normal controls were given a referential communication task structured to assess lexical encoding of information, pragmatic/conceptual elaboration of information and effectiveness in establishing reference. The subjects also received standardised aphasia tests (CADL, Holland, 1980; and Cookie Theft Picture Description, Nicholas and Brookshire, 1993). Comparable reduction of lexical encoding of information was found in the discourse of aphasic and DAT participants both on the referential communication task measures and on the standardised evaluation with the Cookie Theft Picture Description test. However, the DAT subjects' discourse on the referential communication task was less efficient in establishing reference than that of the aphasics since the former presented more misunderstandings and required more explicit prompts from the listener. Furthermore, the DAT language on the referential communication task contained confounding and irrelevant information; also, the number of these errors correlated negatively with their referring abilities. Results of the CADL test confirmed that the DAT participants had less communicative effectiveness than their lexical deficit alone predicted. Finally, examination of the performance of individual DAT subjects showed that lexical encoding of information could dissociate from effectiveness in making reference. These findings support the view that difficulty in pragmatic/conceptual elaboration of discourse information content plays a substantial role in the development of reduced information content and lack of reference of DAT "empty speech". These results are discussed in the framework of the hypothesis of early attentional/executive impairment in DAT (Perry and Hodges, 1999). PMID- 16042029 TI - Coverbal gestures in Alzheimer's type dementia. AB - Patterns of conversational gestures were analysed in subjects with Alzheimer's type dementia (DAT), fluent aphasics with a primarily lexical-semantic deficit (FA) and normal subjects. The FA subjects produced twice as many gestures as the normal participants with a normal percentage of gestures that showed semantic features of the lexical items in concurrent speech (iconic). A comparable lexical semantic deficit together with a deficit in conceptual organisation of information corresponded to a normal gesturing rate in the DAT subjects; however, the percentage of iconic gestures was reduced. Gestures were also analysed in four DAT patients whose communicative performance indicated primarily lexical semantic (2 patients) or conceptual deficit (2 patients). In the two DAT patients with lexical-semantic deficit, the gesture pattern was like that of the FA patients; in the other two, the pattern of the DAT group was observed. These results agree with previous findings that DAT "empty" speech corresponds to reduced production of gestures showing semantic features (Glosser et al., 1998). However, the comparison between DAT with primarily lexical-semantic or conceptual deficits indicates that the nature of the cognitive impairment underlying poor information content and lack of reference in DAT discourse constrains the production of conversational gestures by patients with this disease. These findings are at variance with the hypothesis of parallel dissolution of speech and gestures in language disorders after brain damage (Cicone et al., 1979; McNeill, 1992; Glosser et al., 1998). PMID- 16042030 TI - Egocentric disorientation following bilateral parietal lobe damage. AB - Aguirre and D'Esposito (1999) suggested a taxonomy and theoretical framework for understanding topographical disorders. One of the problems they described involved egocentric disorientation, in which deficits are not strictly confined to the topographical sphere but are seen on a wide variety of visuo-spatial paradigms. Here, we report a neuropsychological investigation of MU, a person with egocentric disorientation. To test the usefulness of Aguirre and D'Esposito's framework, we administered tests which were predicted to be easy or difficult for people with egocentric disorientation to show that MU was impaired on tasks sensitive to egocentric disorientation and that he showed adequate performance on tests sensitive to other types of topographical representation. Thus MU showed normal performance on a test of recognition of famous landmarks and he could identify photographs of personally familiar places in his home town, yet he could not say how to get from a recognised building to another place in his environment. His performance fulfils the criteria for egocentric disorientation and fits the predictions derived from Aguirre and D'Esposito's views. PMID- 16042031 TI - Anomic alexia of kanji in a patient with anomic aphasia. AB - The ability to read aloud kanji (logogram) words and to comprehend their meaning was systematically examined to clarify the underlying mechanism of kanji alexia in a patient with anomic aphasia. Confrontation naming, reading aloud and reading comprehension tasks were performed using 110 words from 11 semantic categories written in kanji or kana. Performance in oral reading of kanji words was significantly worse than oral reading of the same words transcribed into kana words. In addition, for kanji words reading aloud was much worse than reading comprehension. Oral reading of kanji words had a significant correlation with naming pictures corresponding to the words, but no correlation with comprehension of kanji words. Qualitative analyses demonstrated that errors in oral reading and naming tasks had many features in common. Our results indicated that some common mechanisms underlie both naming and oral reading of kanji words. We propose calling this type of alexia "anomic alexia of kanji", which should be distinguished from kanji alexia with difficulty in both reading aloud and comprehension. Lesions in our patient were located in the middle part of the left middle temporal gyrus and its subcortical area, which could be important for access to the phonological lexicon from semantics. PMID- 16042032 TI - Age, sex and regional brain volumes predict perceptual-motor skill acquisition. AB - Neural mechanisms of skill acquisition across the lifespan are largely unknown. Neuroimaging in younger adults and research on neurodegenerative disorders suggests that perceptual-motor learning relies on the striatum, cerebellum, and prefrontal cortex. Our objective in this study was to explore whether individual differences in regional brain volumes modify the effects of age and sex on the acquisition of a perceptual-motor skill in healthy adults. The participants (N = 85, age 22-80) performed five 5-trial blocks of a mirror-drawing task on three separate days (for a total of 25 trials). Index of performance was time to completion and number of errors committed. All participants improved with practice, but younger adults performed better than their older counterparts, and women performed better than men. Four brain regions--lateral prefrontal cortex (LPFC), hippocampus (HC), cerebellar hemispheres (Cb) and the caudate nucleus (Cd)--were selected on the basis of the relevant literature and measured on MRI scans. All regional volumes were negatively associated with age, although the magnitude of association differed from LPFC (the strongest) to Cd (the weakest). Larger lateral PFC was associated with better performance on the mirror drawing task, and this link was stronger in the older participants and was strengthened at the later stages of learning. Larger caudate was related to better performance, especially at later learning, among men, but among women the link was evident only during early learning. Thus, mirror-drawing represents a task that, despite its visual-spatial nature, favors women, and may exhibit sexually dimorphic brain substrates. PMID- 16042033 TI - Adjustments of cognitive control in younger and older adults. AB - The effects of aging on behavioral and electrophysiological indices of adjustments of cognitive control were examined in two experiments. Specifically, we considered the effects of aging of patterns of response time and modulations of the event-related brain potentials (ERPs) on sequential trial effects, error related slowing, and local switch costs in two modified Stroop tasks. The behavioral data reveal that sequential trial effects were observed when color, but not word, identification was required and that these effects were similar in younger and older adults; that the degree of error-related slowing was similar for younger and older adults in Experiment 1 and greater in older than younger adults in Experiment 2; that local switch costs in response time were similar in younger and older adults and that the requirement to switch between color and word identification resulted in a dramatic increase in the number of color intrusion errors in older adults in Experiment 2. The ERP data revealed that endogenously generated adjustments of cognitive control were associated with a parietal slow wave that mirrored the behavioral data and was similar in amplitude in younger and older adults and an anterior frontal slow wave that was absent in older adults. The ERP data also revealed that an enhancement of the P3 component and a frontal slow wave that differentiated color switch trials from color non switch and word trials in younger adults were attenuated in older adults. PMID- 16042034 TI - Dissociating age-related changes in cognitive strategy and neural efficiency using event-related fMRI. AB - We used event-related fMRI to measure brain activity while younger and older adults performed an item-recognition task in which the memory-set size varied between 1 and 8 letters. Each trial was composed of a 4-second encoding period in which subjects viewed random letter strings, a 12-second retention period and a 2 second retrieval period in which subjects decided whether a single probe letter was or was not part of the memory set. For both groups, reaction time increased and accuracy decreased with increasing memory set-size. There were minimal age related differences in activation patterns with increasing memory set-size in prefrontal cortex (PFC). Regression analyses of individual subjects' performance and cortical activity indicated that speed and accuracy accounted for considerable variance in dorsal and ventral PFC activity during encoding and retrieval. These results suggest that younger and older adults utilize similar working memory (WM) strategies to accommodate increasing memory demand. They support a model of cognitive slowing in which processing rate is related to neural efficiency. PMID- 16042035 TI - Relating medial temporal lobe volume to frontal fMRI activation for memory encoding in older adults. AB - Neuroimaging research on the brain basis of memory decline in older adults typically has examined age-related changes either in structure or in function. Structural imaging studies have found that smaller medial temporal lobe (MTL) volumes are associated with lower memory performance. Functional imaging studies have found that older adults often exhibit bilateral frontal-lobe activation under conditions where young adults exhibit unilateral frontal activation. As yet, no one has examined whether these MTL structural and frontal-lobe functional findings are associated. In this study, we tested whether these findings were correlated in a population of healthy older adults in whom we previously demonstrated verbal memory performance was positively associated with left entorhinal cortex volume in the MTL (Rosen et al., 2003) and right frontal lobe activation during memory encoding (Rosen et al., 2002). Thirteen, non-demented, community-dwelling older adults participated both in a functional MRI (fMRI) study of verbal memory encoding and structural imaging. MRI-derived left entorhinal volume was measured on structural images and entered as a regressor against fMRI activation during verbal memory encoding. Right frontal activation (Brodmann's Area 47/insula) was positively correlated with left entorhinal cortex volume. These findings indicate a positive association between MTL volume and right frontal-lobe function that may underlie variability in memory performance among the elderly, and also suggest a two-stage model of memory decline in aging. PMID- 16042036 TI - Alzheimer's disease and depression: preclinical comorbidity effects on cognitive functioning. AB - Both Alzheimer's disease (AD) and depression (D) are prevalent disorders in old age and may co-occur in the same individual. The present study examined whether a diagnosis of D in AD has negative effects on cognitive functioning in the preclinical stage of the diseases, as well as at the time when the diagnoses were rendered. Population-based samples of 13 individuals with incident AD and D, 109 incident AD cases without D, and 179 normal older adults were followed over a three-year period. The groups were compared preclinically and at the time of diagnosis on global cognitive functioning using the MMSE total and the specific item scores, as well as the occurrence of depressive symptoms. As expected, there were clear AD-related deficits preclinically, which were exacerbated at follow up. In addition, there were D-related deficits on three MMSE items (i.e., following commands, reading, and writing). The poorer performance on the three MMSE items was linked to an elevation of depressive symptoms. However, D was not associated with greater decline in cognitive functioning over the three-year follow-up period. Thus, although depressive symptoms may result in slight cognitive deficits in preclinical AD, at the time of the dementia diagnosis these effects may be absorbed by the neurodegenerative process. PMID- 16042037 TI - Pilot study of the Olympia oyster Ostrea conchaphila in the San Francisco Bay estuary: description and distribution of diseases. AB - Olympia oysters Ostrea conchaphila have declined markedly during the last century and are a focus of restoration in many embayments, including the San Francisco Bay (SFB) estuary. Oysters were collected from 17 sites in this estuary and nearby Tomales Bay in an effort to characterize diseases that may impact recovery of this species and captive rearing programs. Three diseases/disease agents including a Mikrocytos-like protist (microcell), a haplosporidian and hemic neoplasia were observed from several sites along the western margins of the SFB estuary suggesting a geographic localization of disease presence. Based on fluoresecent in situ hybridization (FISH) assays, the microcell is distinct from M. mackini and Bonamia spp. These data highlight the need for further elucidation of the haplosporidian and for careful health management of a declining species destined for captive rearing and supplementation. PMID- 16042038 TI - Validation of in situ hybridisation and histology assays for the detection of the oyster parasite Marteilia refringens. AB - An in situ hybridisation technique has been developed for the detection of infection in oysters with Marteilia refringens with particular emphasis on light infections or confirmation of suspected cases by means of histology. Although validation of new diagnostic methods is usually achieved by comparison with standard techniques, in our case the sensitivity and specificity of the standard (histology) had not previously been established. Another point to consider is that surveillance and monitoring frequently target populations displaying different levels of prevalence under different field conditions. The objective of our study was to evaluate the sensitivity and specificity values of in situ hybridisation and histology for the detection of M. refringens, based on 3 populations of flat oysters, free of the disease and with mild and high levels of prevalence. A blind assay of 200 individuals from each population was performed using both techniques. Results were analysed by means of the classical approach and latent models (maximum likehood and Bayesian approach). Assumptions and results were found to vary slightly with the different statistical approaches. The more realistic estimate by the Bayesian approach shows a link between the level of prevalence and the sensitivity of the techniques. Values of sensitivity and specificity for histology were 0.7 and 0.99 respectively, and 0.9 and 0.99 respectively in the case of in situ hybridisation. Some uncertainty remains regarding these values because the study does not take into account the severity of infection or the developmental stages of the parasite actually present in each individual. This work provides valuable information with regard to the choice and potential use of those 2 diagnostic methods currently recommended by international standards. PMID- 16042039 TI - Monoclonal antibodies developed for detection of an epizootic virus associated with mass mortalities of cultured scallop Chlamys farreri. AB - Recently, an enveloped, spherical RNA virus was identified as the causative agent of mass mortalities among adult scallop Chlamys farreri, which is cultured on the northern coast of China. Hybridomas were prepared from mice immunized with highly purified virions. Four stable hybridomas secreting monoclonal antibodies (MAbs) of IgG isotype were obtained after screening by means of enzyme-linked immunosorbent assay (ELISA) and immunofluorescence assay (IFA). The specificity of the MAbs to this virus was confirmed by immunogold electron microscopy (IEM). All the selected MAbs recognized epitopes on the envelope spikes of the virions. Subsequently, the MAbs were used for in situ immunofluorescent detection of the virus in Davidson's fixed tissue sections. The results showed that the fluorescent cells were mostly observed in epithelia of different organs, but not in the epithelium of the digestive diverticulae. Cytopathological changes and focal lesions corresponding to virus-positive cells were clearly recognized in the affected epithelia, revealing a potential role of this virus in pathogenesis. PMID- 16042040 TI - Herpes-like virus infection causing mortality of cultured abalone Haliotis diversicolor supertexta in Taiwan. AB - A herpes-like virus is demonstrated for the first time to be associated with high mortality rates in maricultured abalone Haliotis diversicolor supertexta in Taiwan. Histopathology of moribund abalone indicated that the nerve system was the primary target tissue. The lesions were characterised by tissue necrosis accompanied with infiltration of haemocytes. Electron microscopic examination demonstrated viral particles within the degenerated cerebral ganglion cells. The viruses were hexagonal, approximately 100 nm in diameter and had a single coat. Some viral particles contained a dense nucleoid, while others were empty. The ultrastructure and morphogenesis of the virus particles were consistent with those of the herpesvirus described from the oyster Crassostrea virginica. Experimental infection using supernatant collected from minced visceral organs and muscle of moribund abalone induced 100 % mortality through both intramuscular injection and bath treatments. PMID- 16042041 TI - Immunosuppression in progeny of chinook salmon infected with Renibacterium salmoninarum: re-analysis of a brood stock segregation experiment. AB - Female spawner infection level and temperature variation through rearing are sufficient to explain in-hatchery mortality rates and infection levels and smolt to adult return ratios (SARs) of progeny of Renibacterium salmoninarum infected spring chinook salmon. Data from published reports and manuscripts regarding a 1988 brood stock segregation experiment that held progeny of highly infected female spring chinook salmon spawners separate from progeny of other spawners during 16 mo of hatchery rearing are analyzed to test the hypothesis that immunosuppression could account for differences in survival and infection levels between the 2 segregates. Immunosuppression, caused by the presence of the p57 antigen of R. salmoninarum in sufficient concentration within the salmon egg before spawning, can account for differences in infection levels, mortality rates, and SARs for each hatchery raceway in that study. This immunosuppression may be characterized by immunotolerance, or might only affect cell mediated immunity, which appears the most effective defense mechanism against R. salmoninarum infection, as antibody production can result in tissue damaging antibody-antigen complexes. Low-temperature mediated immunosuppression can account for the nearly identical trajectories of infection and mortality between the 2 segregates during the first 8 mo of hatchery rearing. There is no evidence of widespread vertical infection from spawner to progeny, nor is there evidence that brood stock segregation reduces overall mortality. Rather, the suppression of cell-mediated immune mechanisms may condemn progeny of highly infected female spawners to an almost certain eventual premature demise. PMID- 16042042 TI - Quorum sensing signal molecules (acylated homoserine lactones) in gram-negative fish pathogenic bacteria. AB - The aim of the present study was to investigate the production of quorum sensing signals (specifically acylated homoserine lactones, AHLs) among a selection of strains of Gram-negative fish bacterial pathogens. These signals are involved in the regulation of virulence factors in some human and plant-pathogenic bacteria. A total of 59 strains, representing 9 different fish pathogenic species, were tested against 2 AHL monitor bacteria (Agrobacterium tumefaciens NT1 [pZLR4] and Chromobacterium violaceum CV026) in a well diffusion assay and by thin-layer chromatography (TLC). Representative samples were further characterized by high performance liquid chromatography-high resolution mass spectrometry (HPLC-HR-MS). AHLs were produced by all strains of Aeromonas salmonicida, Aeromonas hydrophila, Yersinia ruckeri, Vibrio salmonicida, and Vibrio vulnificus. Some strains of atypical Aeromonas salmonicida and Vibrio splendidus were also positive. Aeromonas species produced N-butanoyl homoserine lactone (BHL) and N-hexanoyl homoserine lactone (HHL) and 1 additional product, whereas N-3-oxo-hexanoyl homoserine lactone (OHHL) and HHL were detected in Vibrio salmonicida. N-3-oxo octanoyl homoserine lactone (OOHL) and N-3-octanoyl homoserine lactone (OHL) were detected in Y. ruckeri. AHLs were not detected from strains of Photobacterium damselae, Flavobacterium psychrophilum or Moritella viscosa. AHLs were extracted from fish infected with Y. ruckeri but not from fish infected with A. salmonicida. In conclusion, the production of quorum sensing signals, AHLs, is common among the strains that we examined. If the AHL molecules regulate the expression of the virulence phenotype in these bacteria, as shown to occur in some bacterial pathogens, novel disease control measures may be developed by blocking AHL-mediated communication and suppressing virulence. PMID- 16042043 TI - Direct identification of Photobacterium damselae subspecies piscicida by PCR-RFLP analysis. AB - Fish pasteurellosis is an infectious disease that affects several teleost species living in temperate marine waters. The pathogen responsible, Photobacterium damselae subspecies piscicida, shows high genetic similarity with P. damselae subsp. damselae, making subspecies discrimination extremely laborious. Here we report for the first time a PCR-RFLP method for the identification of P. damselae subsp. piscicida without prior isolation in pure culture. Genomic sequence information was obtained through cloning and sequencing of RAPD products. Two P. damselae-specific primer pairs were developed and tested on 17 strains of P. damselae subsp. piscicida, 10 strains of P. damselae subsp. damselae, and 6 closely related control species. High sensitivity was achieved in PCR amplification on serially diluted samples (<180 fg of pure bacterial DNA or <10 fg, depending on the amplified fragment). Restriction analysis of PCR products showed a unique digestion profile for all P. damselae subsp. piscicida strains. The same PCR-RFLP method was implemented on total DNA samples extracted from experimentally infected sea bream and sea bass. Positive results were obtained on fish with clear signs of the disease as well as on challenged, but asymptomatic, fish. The method presented here might provide a useful tool for both prevention and rapid diagnosis of fish pasteurellosis. PMID- 16042044 TI - Blood protozoans in elasmobranchs of the family Rajidae from Galicia (NW Spain). AB - Blood smears from 132 skates Raja spp. captured on the continental shelf off Galicia (NW Spain) were examined for blood parasites. The skate species were Raja brachyura (n=60), R. microocellata (n=52) and a total of 20 specimens belonging to R. undulata, R. montagui and another 2 unidentified Raja species, all captured between March 1999 and March 2000. Two blood parasite species were found, Trypanosoma giganteum and Haemogregarina delagei. Of the 132 skates, 16% were infected only by T. giganteum, 17% only by H. delagei, and 5% by both T. giganteum and H. delagei. Both parasites showed highest prevalence in R. brachyura (22% T. giganteum only, 38% H. delagei only, 12% T. giganteum and H. delagei). Mean leucocyte percentages (n=132 fish) were lymphocytes (43%), eosinophils (35%), neutrophils (20%) and monocytes (2%); basophils were not found. As far as we are aware, this eosinophil percentage is the highest reported to date for elasmobranchs. We did not detect any statistically significant differences in leucocyte percentages between infected and uninfected fish, between male and female fish, among species or among weight groups. PMID- 16042045 TI - Population dynamics of the philometrid nematode Margolisianum bulbosum in the southern flounder Paralichthys lethostigma (Pisces: Paralichthyidae) in South Carolina, USA. AB - This is the first report of the philometrid nematode Margolisianum bulbosum Blaylock and Overstreet, 1999 from the southern flounder Paralichthys lethostigma on the east coast of the USA. Observation of adult female worms was used as an indication of the parasite's presence in the fish. Adult females were found only in P. lethostigma >50 mm total length. The overall prevalence was 74%, with a mean intensity of 5 female nematodes per parasitized fish. Infected flounders were found throughout the year with a statistically significant decrease in intensity in the winter months. Neither salinity, water temperature, fish gender nor fish age were found to influence either prevalence or intensity of infection in the flounder. While larvigerous (gravid) females were found throughout the year, the significant decrease in their occurrence during the summer through fall, in concert with an observed decrease in intensity of infection during the winter, indicated that the life cycle of this philometrid species is likely to be annual. PMID- 16042046 TI - Effect of chronic Taura syndrome virus infection on salinity tolerance of Litopenaeus vannamei. AB - Taura syndrome virus (TSV) is one of the most important shrimp viruses affecting farmed shrimp worldwide. After an acute phase during which the likelihood of mortality is elevated, infected shrimp enter a chronic phase during which shrimp appear to resume normal behavior and display no gross signs of infection. This study was designed to determine if chronically TSV-infected shrimp Litopenaeus vannamei are compromised by the infection. Specifically we investigated whether chronically infected shrimp could tolerate a drop in salinity as strongly as uninfected shrimp. The study consisted of 3 trials that compared survival of uninfected and chronically TSV-infected L. vannamei after drops in salinity from 24 ppt to salinities varying from 18 to 0 ppt. Logistic regression detected a significant effect of TSV infection on survival of chronically infected shrimp (p < 0.05). Salinity drops from 24 ppt to 3 and 6 ppt resulted in statistically different survivals (p < 0.05). Survival rates were similar among groups for salinity drops to greater than 6 ppt or less than 3 ppt. Salinities at which 50% of the shrimp died (LC50) were 3.06 ppt for the uninfected and 6.65 ppt for the chronically infected groups. Moreover, histopathological analysis of chronically infected shrimp that were moribund or recently dead showed no signs of having reverted to the acute stage of the disease. These results suggest that chronically infected shrimp are not able to tolerate a salinity drop as strongly as uninfected shrimp. PMID- 16042047 TI - Synthethic peptide used to develop antibodies for detection of polyhedrin from monodon baculovirus (MBV). AB - The use of previously published primers to amplify the monodon baculovirus (MBV) polyhedrin gene sequence by polymerase chain reaction (PCR) from post larvae (PL) of Thai Penaeus monodon resulted in failure. As a result, the putative polyhedrin protein of MBV was isolated from infected PL by homogenization, differential centrifugation and density gradient centrifugation with verification by transmission electron microscopy (TEM). By SDS-PAGE, a single major protein band at 58 kDa was obtained from the putative polyhedrin fraction and this corresponded to a previous report of the molecular weight of polyhedrin from MBV. When used for N-terminal sequence analysis, the putative polyhedrin protein yielded a sequence of 25 amino acids (M F D D S M M M E N M D D L S G D Q K M V L T L A) that did not correspond to the deduced amino acid sequence derived from a previous report of a putative MBV polyhedrin gene amplicon. Despite this, a synthetic peptide of our 25 amino acid sequence (25Pmbv) was conjugated with bovine serum albumin and used as an antigen for antiserum production in mice. Using immunohistochemistry with tissue sections of PL infected with MBV or other viruses, the mouse anti-25Pmbv antiserum showed strong immunoreactivity to occlusion bodies of MBV only. It also showed strong reactivity to the 58 kDa putative polyhedrin protein in Western blots. Altogether, the results suggest that the 58 kDa protein is Thai MBV polyhedrin and that a previously reported MBV polyhedrin gene sequence may represent another protein or polyhedrin from a different variety of MBV. PMID- 16042048 TI - Generation of monoclonal antibodies specific to Hepatopancreatic parvovirus (HPV) from Penaeus monodon. AB - Hepatopancreatic parvovirus (HPV) was isolated from the hepatopancreas (HP) of slow growth Penaeus monodon by urografin gradient centrifugation. The presence of HPV in the fraction was monitored by PCR and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Only 1 major 54 kDa protein band was observed in the strong PCR-positive fractions used to immunize mice for monoclonal antibody production. After cell fusion, the first step in selecting specific antibodies was performed by dot-blot assay with purified HPV viral particles. The second screening step was carried out using Western blots of purified HPV proteins and immunohistochemistry of HPV-infected HP tissue. Four monoclonal antibodies were isolated; these bound to the 54 kDa protein in Western blots and to intranuclear inclusion bodies in tubule epithelial cells of HPV infected prawn tissue by immunohistochemistry. None of the antibodies showed cross-reactivity either to uninfected shrimp tissue or to other shrimp viruses tested. These reagents have potential for use in developing a highly sensitive immunoassay such as sandwich ELISA or a convenient kit for detection of HPV infection. PMID- 16042049 TI - Hereditary spherocytosis is more frequent than expected: what to tell the patient? AB - Modern double beam laser technique allows screening for hereditary spherocytosis in the course of routine hematology. An incidence of 1:150 men and 1:800 women has been determined. The anomaly is symptomless in the majority of the cases. This explains the discrepancy between our values and the incidence of 1:5,000 reported in the literature. The diagnosis of hereditary spherocytosis should be reported to the physician and the patient, as it may be wayleading in case of unexpected, unspecific complications such as anemia, jaundice, cholelithiasis, liver cell damage and iron overload. Regular monitoring of plasma ferritin and glucose is recommended. PMID- 16042050 TI - [Alcohol, pharmaceuticals, illicit drugs and driving]. AB - A great number of clinical, epidemiological, pharmacological and toxicological data on the influence of psychotropics on driving are available. These psychotropics include psycholeptics like ethanol, opioids, psychoanaleptics like cocaine, amphetamines and congeners, psychodysleptics like cannabis, LSD and magic mushrooms. General epidemiology and specific epidemiology for Luxembourg will be outlined. Practical aspects of roadside testing, forensic aspects as well as the place of hair testing in drugs and road safety issues will be discussed. PMID- 16042051 TI - [Limitations and possibilities of applying the positive effects of human/animal relations in the treatment of psychological ailments]. AB - For the 72nd Psychiatric Saar-Lorr-Lux Symposium (Sept. 2004 at the Neuropsychiatric Hospital (CHNP) in Ettelbruck, LUXEMBOURG), Dr. Jean-Marie Spautz, director, brought together professionals of varied disciplines, each involved in Animal Assisted Therapy (AAT). PMID- 16042052 TI - [Dogs and people]. AB - A neuropsychiatrist remembers all the dogs he met in his life. Ten of them, with different characters, their own ways to express affectionate wishes by continuous body language. Not only patients are helped by animals who are their friends but so are doctors. PMID- 16042053 TI - [Increase in the wealth of emergency patients at the Central Hospital of Luxembourg. Who are they? Why do the come?]. AB - As in all European countries, the CHL's Emergency Department is faced with a constant increase in the number of patients. The demographic patient profile in the 2001 survey shows a majority of young males with traumatic problems. Mostly evoked reasons for the increase are the lack of availability of general practitioners and patients' low level of information about the competences of GPs. The article discusses possible solutions to the bottlenecks in Emergency Departments. PMID- 16042054 TI - [Therapeutic and microbiologic aspects of pleural empyema]. AB - The pleural empyema is a severe disease which passes with respiratory insufficiency and general toxic symptoms. The early staged surgical treatment combined with total evacuation of the puss from pleural cavity is important for the successful outcome of the disease. We made retrospective study including 82 patients with pleural (61 with acute and 21 with chronic) empyema. We performed the following operative interventions: thoracocentesis with active aspiration--at 43, VATS--20, decortication and pleurectomy--in 16, thoracoplastic with muscle flap--3. Three patients died. The most common causes for the empyema were Gr (+) coccy. At most of the patients combined operative treatment and antibiotic therapy were the most effective. PMID- 16042055 TI - [Surgical treatment of chronical pulmonary abscesses--contemporary treatment]. AB - Between Jan 1991 and Dec 2000 a total of 67 patients were operated on for chronic primary pulmonary abscesses. They were 57 men and 10 women with mean age of 46.2 (range, 12 to 72) years. The indications for surgery was mainly "delayed closure" following medical therapy and percutaneous tube drainage (3), as well as a massive hemoptysis in 4 cases. CT scanning and brochoscopy were performed in all patients. The majority of them had multiple isolates (51/67) of both aerobic and anaerobic (predominantly Bacteroides sp., Fusobacterium sp., and Peptococci) organisms. The following operations were carried out: lobectomy (52) including 8 decortications, bilobectomy (5), pneumonectomy (4) including two pleuro pneumonectomy, polysegmentectomy (4) and segmental resection (2). The 30-day hospital mortality rate was 1.49% (1 patient died following rethoracotomy for bronchial stump fistula with empyema and polyorganic insufficiency). Major postoperative complications were 6 (8.9%) and included 3 pleural empyema (additional drainage), two rethoracotomy for intrapleural bleeding and one residual pleural cavity, treated by thoracoplasty. Minor postoperative complications (atelectasis, wound infection and prolonged air-leakage) were observed in 9 patients (13.4%). The long-term results (following-up ranged from 6 to 112 weeks) are considered very good. In conclusion, surgery is indicated for patients with significant hemoptysis, suspected malignancy and those with "late healing" abscesses with acceptable postoperative results, although the rate of major postoperative complications remains relatively high. PMID- 16042056 TI - [Videoassisted thoracoscopic treatment of retained fluid collection in the pleural cavity after thoracotomy]. AB - INTRODUCTION: One of the common complications of the thoracotomies is the formation of incapsulated fluid collections in the pleural cavity. There are different invasive methods for their treatment. MATERIALS AND METHODS: The authors shared their experience with the videoassisted thoracoscopic treatment of 19 patients with pleural collections, treated in the clinic for a period of four years (2000-2003 year). RESULTS: VATS at 17 of the patients leads to healing. We performed rethoracotomy at two of the patients because there were no effect of VATS. CONCLUSION: VATS is an alternative of rethoracotomy in the operative treatment of postoperative fluid collections in the pleural cavity. PMID- 16042057 TI - [Papillary and cystic tumor of pancreas--Frantz's tumor]. AB - The aim of the authors is to introduce one rare known pathology which the Frantz's tumor is, his clinical characteristic, clinical signs, diagnostic and treatment. We have observed two separate cases of this illness in Clinic of Surgery (University Hospital "Queen Yoanna") in comparison with evidence of famous world surgeons. We have made an attempt to introduce one disease, which even though not often observed is significant by the fact that affect young women (girls) and only surgical resection is a treatment of choice. PMID- 16042058 TI - [Micrometastases in patients with colorectal cancer: markers for metastatic potential or proofs for residual disease]. AB - It is know that bone marrow micrometastases are an indicator of poor prognosis in patients with epithelial tumors. At the same time little is known if they reflect minimal residual disease or cells distribution and marked metastatic potential. We aimed to perform a clinical study, which though with little gathered clinical material has a direction--investigation of bone marrow in patients with colorectal cancer by the time of surgery, carefully rendering an account of staging and performing curative resection. In intraoperatively done bone marrow biopsy with following investigation for cytoceratin 19 positive cells we found ol presence of metastatic cells in some the studied patients. Finding micrometastases in patients with colorectal cancer is a marker for more agressive course of the disease and for the need to change some of recent methods of treatment. PMID- 16042059 TI - [Uroinfections and the possibilities of therapy with levofloxacin]. AB - The common occurrence, of inflammatory diseases in the urinary tract and the genitals in men, is quite a reason for identification of the agents and looking for some new chances of therapeutic treatment. MATERIALS AND METHODS: For a period of two years (2002-2003) we accomplished 4810 microbiological tests of urine and prostate secretion on patients with uroinfections. The cultures are completed on standard nutrition medium, as the biochemical identification of the strains is accomplished by the systems mini API and "bio Merieux" BBL Crystal, "Becton Dickinson". Meanwhile for a period of one year, 113 patients with different uroinfections are treated and studied. In 16 (14.1%) Levofloxacin (Tavanic-R) is applied i.v. 500mg/daily as in the rest cases 97 (85.9%) Tavanic are applied per oral in different therapeutic doses. RESULTS AND DISCUSSIONS: The most common agent in our study was the E. Coli, as for the year 2003 we read an increase of the infections caused by E.Coli with 3.4% comparing to the previous one (38.5% to 35.1%). The differentiation of patients treated with Levofloxacin in groups is completed as follows: with acute uncomplicated cystitis- pyelonephritis after ESWL--15.9%; with postoperative uroinfections--20.3%; with chronic post-tuberculosis pyelonephritis--4.4%; and with inflammatory diseases of the male genitals--37.2%. PMID- 16042060 TI - [Surgical treatment in complications of renal transplantation]. AB - The increasing number of renal transplantations in Bulgaria and the increased survival rate of renal transplantation patients lead to an increase of the number of complications (vascular, urological, liquid collections) requiring surgical intervention. One hundred and ninety renal transplantation patients have been treated in the Department of Renal Transplantation for the period 1997-2003. Of these 12 were admitted to the Clinic of Vascular Surgery. We observed 36 surgical complications: arterial thrombosis--7, venous thrombosis--1, arterial stenosis- 7, lymphocoele--4, hematoma--4, ureteral fistula--6, ureteral stricture--6, ureterolithiasis--1. We undertook following surgical interventions: vascular reconstruction, thrombectomy, lymphocoele punction, fenestration towards peritoneal cavity, reimplantation techniques (a modo Gregoari, Boeminghausi, Boari), termino-terminal anastomosis with native ureter, uretero-enteric anastomosis, ESWL of ureteral stone. Renal transplantation complications requiring surgical interventions are serious urological problem. The severity, the type (vascular, liquid collection, urological), and the diagnosis of these complications determine patients' prognosis. Urgent surgical interventions lead to rapid recovery of graft function, eradication of the infection, amelioration of blood flow, and improved graft survival. PMID- 16042061 TI - [Intraoperative effect of preoperative embolization of the renal artery in patients with kidney carcinoma]. AB - Aim of the embolization of the renal artery is to provoke ischemia or shut out from blood supply of the kidney tissue including pathologically changed area. As a result of this procedure the affected kidney becomes suitable for surgical removal without significant loss of blood. In this study there are included 116 nephrectomies due to kidney carcinoma. Clinical diagnosis and treatment of the patients are held at the Clinic of Urology of the Alexandrovska Hospital for the period 1990-1995. Embolization of the renal artery is accomplished in 52 subjects (63.41%). The procedure was contraindicated in 10 subjects (8.62%). It was established intraoperatively that the affected kidney had good embolization in 46 subjects (88.46%), in 4 subjects (3.44%) it had excellent embolization, and in 2 subjects (1.72%) there was just peripheral embolization. Authors conclude that: 1) Nevertheless this method is invasive it still has a place in urologists' practice. 2) Preoperative embolization of the renal artery is a devise for surgical removal of the kidney as well as achieving maximum effect. PMID- 16042062 TI - [LUTS in women and possible therapeutic effects of alfa1-adrenoblockers]. AB - Lower Urinary Tract Symptoms (LUTS), including urinary incontinence, are one of the most common problems in women pathology. OBJECTIVE: The objective of the authors is to determine the symptomatic index, to perform an urodynamic assessment and to establish the therapeutic possibilities of the alfa1 adrenoblokers in women with LUTS. MATERIAL AND METHODS: In the course of a three year period (2001-2003) 74 women, ranging in age from 26 to 76 years old (average age of 48.6), have been examined and monitored. The symptomatic index was measured by a questionnaire, developed on the basis of IPSS and BFLUTS. The urodynamic study was performed with a multi-channel system Dantec-Meneut and includes multi-chanel cystrometry, followed by measuring of the pressure and the speed of urinary flow before the treatment and after it. Therapy with the L1 adrenoblocker Doxazosin was applied to 42 (56.7%) of the patients. RESULTS: 51 (68.9%) women were with predominantly irritative symptoms, and 23 (31.1%)--with predominantly obstructive. The urodynamic studies reval: 1. Obstruction of the lower urinary tract--7 (9.4%); 2. Obstruction with detrusor hyperactivity--15 (20.3%); 3. Detrusor hyperactivity--33 (44.6%); 4. Sensory urge-syndrome--6 (8.1%); 5. Disturbed contractility of the urinary bladder--4 (5.4%); 6. With no significant deviations--9 (12.2%) Total disappearance of the symptoms was achieved in 64.3% of the patients with the prescribed therapy, while in 11.9% symptoms have relapsed after discontinuation of the therapy. CONCLUSIONS: LUTS in women is not only a gerontology problem. It affects more and more often women in active and child-bearing age. The therapeutic use of alfa1-adrenoblockers is one alternatives in the treatment of these patients. PMID- 16042063 TI - [Treatment with cefrom (cefpirom) in severe urological infections]. AB - Severe uro-genital infections frequently complicate a wide range of urological conditions. The favorable outcome of these infections depends mainly on the choice of appropriate antimicrobial treatment. Cefrom is bactericidal beta lactamase-resistant IV generation cephalosporine antibiotic. In 2003 we treated 20 patients with severe urinary tract infections with Cefrom. We achieved excellent therapeutic results--urinary tract infections were eradicated in all patients, as shown by the microbiological investigations of urine specimen after the antibacterial course of treatment. The wide antimicrobial spectrum, the good tissue permeability in the organs of the genito-urinary tract and the good tolerability of Cefrom make this antibiotic a drug of choice in patients with acute and complicated urological infections. PMID- 16042064 TI - [The role of polyclonal anti-T-lymphocyte antibodies (ATG) in the kidney transplantation]. AB - The successes in the kidney transplantation are closely connected with the successes of the immunology and the advance of the immunosuppressive therapy. The main task of the modern immunosuppressive therapy is the insurance of medicines with a minimum nephro-toxicity, and the optimum protection regarding the adoptive body against an early reaction of rejection. The polycomponent anti-T-limphocitic antibodies (ATG) give us the opportunity of that. The polycomponent anti-T limphocitic antibodies or the antilimphocitic serum (ALS) are xenogenetic polycomponent antibodies, that are directed to the human T-limphocites, i.e. antitimocitic globuline (ATG). They are received by immunization of rabbits or horses with human thymus limphoid cells. They are received for the first time by horses, and later on by rabbits. The last patent medicines have more powerful effect. In spite of the side effects of the polycomponent ATG, the patent medicine is used for inductive treatment after transplantation of organs and for a treatment of acutely rejection of the graft. We have treated 15 patients after kidney transplantation with ATG for a period of an year in the ward of kidney transplantation in The Clinic of Urology. The patients with transplantation are at the age of 21 to 50 years old, and by sex--11 male and 4 female patients. 9 of them are transplanted from alive donor and 6--from a dead body. We have applied the patent medicine of ATG Timoglobuline in a phial of 5 mg?ml intravenally in a bank of 500 ml physiological solution according to the weight and the blood test of the patients. We have applied a fourfold immunosuppressive therapy: Urbazon, Imuran, CyA(Neoral) +ATG. CONCLUSIONS: ATG is a mixture of monocomponent polyspecific antibodies, which, in spite of its side effects, has been applied successfully in transplantation for more than 30 years. In the course of many years anti-CD3 patent medicine OKT3 has been the only one monocomponent antibody for the treatment of an acute rejection of the graft for inductive therapy in transplanted patients.A chimerical (baziliksimab) and humanized (daklizumab) monocomponent antibodies of the alpha-chain of the receptor for IL-2 (CD25) are used for the prophylaxis of the episodes of acutely rejection of the transplanted kidney during the last 10 years. ATG does not consists of anti-CD25 antibodies. That is why the simultaneously application of ATG with baziliksimab or daklizumab is expected to have an additional effect with a complementary mechanism of efficacy. PMID- 16042065 TI - [Colorectal localization in Crohn's disease--diagnostics and surgical treatment]. AB - During the last years there is an increase in the number of cases with Crohn's disease. According to different statistic data the localizations are arranged as follows depending on the frequency of occurrence: ileocolic segment localization is the first--41%, followed by colorectal localization--30% and small intestinal one--29%. For a period of five years 12 patients with Crohn's disease, including 7 women and 5 men, were treated in the clinic. Regarding the localization the cases were as follows: 7 in the sigma-rectum transition area, 4 in the cecal area and 1 in the rectal area. All patient were operated and they were in a serious acute form of the disease. The operative intervention, which were made, differed in number: Hartman resection of the sigma--4, sigma resection with primary anastomosis--2; left hemicolectomy--1, right hemicolectomy--3, ileotransverzoanastomosis--1, resectio recti anterior--1. The final diagnosis was given after histological examination. PMID- 16042066 TI - [Surgical treatment of creeping infections of the flexor foot tendons in diabetic patients]. AB - The creeping infections of the flexor foot tendons are a common reason for amputation of limbs in diabetic patients. Usually the process starts from the fingers and spreads through their long flexors. METHODS: Of the total of 149 operated patients, 49 underwent selective removal of the m. flexor hallucis longus tendon, and amputation of the 1st finger. The long flexors of the foot fingers were totally removed through a transmethatarsal amputation in 78 patients, and in 22 the long flexor tendons of the 4 lateral fingers were selectively removed through an oblique foot resection. After this type of surgical operations, limb amputation as a result of losing control over the suppurative-gangrenous process, became necessary in 7 patients. CONCLUSIONS: The infection spreading through the long flexors of the foot fingers in diabetic patients is a serious surgical complication that can only be treated surgically. The method of selective resection and removal of the infected long flexors of the foot fingers allows to avoid the limb amputation in most patients. Following this type of surgical interventions the foot biomechanics and the steadiness of the patient is inconsiderably disrupted. PMID- 16042067 TI - [Surgical treatment of leg arterial occlusions (operative technique)]. AB - The surgical technique of the arterial reconstructions to the leg vessels is of critical importance for the outcome of those delicate procedures. The anatomic surgical exposures to the potentially target leg arteries are described. Infrapopliteal reconstructions are performed when popliteal artery is unsuitable for a distal anastomosis. Posterior tibial artery is exposed through a medial approach as an extension of the bellow knee popliteal artery exposure. Anterior tibial artery is accessible through the medial approach only in its ostial section, distal segments of it can be exposed using antero-lateral approach between tibia and fibula. Peroneal artery can also be exposed using medial approach in its proximal third, and for distal exposure resection of the fibula is required. All patients receive venous antibiotic, and Heparin 100 IU/kg or the corresponding doses low-molecular Heparin. All patients with thrombosed by-passes within 30 days undergo reoperation. Suggested is treatment of skin and soft tissue foot lesions. "Failing graft" should be diagnosed and treated properly, since assisted primary patency is higher than secondary patency. Intraoperative fluoroscopy has the potential to provide valuable information before, during and at completion of the procedure. Mortality is mainly due to myocardial infarction. PMID- 16042068 TI - [Radical operative treatment (nephrectomy and nephroureterectomy) in urology. Aspects and opinions]. AB - The review of literature offerred discusses the opinions of different authors about indications and contraindications for radical oncological procedures in cases of renal and ureteral diseases. PMID- 16042069 TI - [Spontaneous stercoral perforation of colon and rectum with contribution of five cases]. AB - The authors report five cases of spontaneous perforation of the large bowel for a five year period. The clinical symptoms were dominated by the feculent peritonitis. There were found no data of previous diseases of the colon during the operations and hystologically. By all five patients was manifested severe constipation before the incident. During the operation the recto sigmoid and the left colon were found plugged by stercolithes with "stone density". A hypothesis of stercoral pathogenesis of the perforations is suggested. Conclusions for the surgical tactics in the conditions of large bowel peritonitis due to benign perforation are done. PMID- 16042070 TI - Synthesis and anti-inflammatory testing of some new compounds incorporating 5 aminosalicylic acid (5-ASA) as potential prodrugs. AB - This work includes the synthesis of 15 final compounds (6a-h and 7b-h) as prodrugs of 5-ASA in the form of the acid itself, esters and amides linked by an amide linkage through a spacer to the endocyclic ring N of nicotinamide. Also, 15 new intermediate compounds were prepared. The target compounds (6b, 6f, 7b, and 7e-h) revealed potent analgesic and anti-inflammatory activities in comparison to sulfasalazine and 5-ASA. In addition, ulcerogenicity, LD50, in vivo and in vitro metabolism of compound 7f were determined. PMID- 16042071 TI - Synthesis of C4-modified acanthoic acid analogs and their biological evaluation as nitric oxide inhibitors. AB - Synthesis and biological evaluation of a series of C4-modified acanthoic acid analogs are reported. Among them, the analog 7 and 10 exhibit potent cellular inhibitory activity in NO inhibition assay. PMID- 16042072 TI - Synthesis and iNOS inhibitory activities of thioflavones. AB - A number of thioflavones has been synthesized and evaluated for their iNOS inhibitory activities. Thiowogonin (6) was obtained from naturally occurring chrysin in 5 steps. Other thioflavones were prepared from the corresponding flavones in a single step by the reaction with Lawesson's reagent. The biological activities of thioflavones were not enhanced by the functional group conversion from carbonyl to thiocarbonyl. Compounds 11 and 13 showed potent NO inhibitory activity at high concentration (40 uM), leading to the possible development of novel neuroprotective agents based on wogonin. PMID- 16042073 TI - Solanoflavone, a new biflavonol glycoside from Solanum melongena: seeking for anti-inflammatory components. AB - A new biflavonol glycoside named as solanoflavone (1) was isolated from aerial part of Solanum melongena. The chemical structure was elucidated as isorhamnetin 3-O-beta-D-glucopyranoside-(4'-->O-->4''')-galangin-3''-O-beta-D-glucopyranoside on the basis of physicochemical and spectroscopic methods, including 2D NMR spectral techniques. PMID- 16042074 TI - Identification of Lactobacillus ruminus SPM0211 isolated from healthy Koreans and its antimicrobial activity against some pathogens. AB - The intestinal microbiota are important to the host with regard to resistance they impart against bacterial infections and their involvement in mediating metabolic functions. Lactic acid producing bacteria such as Lactobacillus play an important physiological role in these matters. The aim of the present study was to isolate Lactobacillus sp. that inhibits enteric pathogens. Initially, 17 isolates from healthy Koreans were collected on Lactobacillus selective medium. Resistance of the isolates to antibiotics including rifampicin, streptomycin, clindamycin and vancomycin was measured. One of the isolate was identified as Lactobacillus ruminus on the basis of bacterial cell morphology, cultural characteristic and biochemical characteristics, 16S rRNA sequence analysis and PCR-RAPD. Antimicrobial activity of the bacterium against Vancomycin Intermediate Resistant Staphylococcus aureus (VISA) and Vancomycin-Resistant Enterococci (VRE) was measured. About 10(4) cells of VISA or VRE were mixed with 1, 5, and 9 mL of L. ruminus SPM 0211 and the final volume was adjusted to 10 mL with brain heart infusion (BHI) broth. The cell suspension was incubated for 3, 6, 9, and 24 h, serially diluted and then plated on BHI agar plates. As numbers of L. ruminus SPM 0211 were increased, viable cell count of VISA and VRE decreased. The strongest antimicrobial activity of SPM 0211 was observed after 9 h incubation in any mixture, almost completely inhibiting the growth of these two bacteria. The results suggest that the freshly isolated L. ruminus SPM 0211 may be used as a pro-biotic microbe that prevents the colonization of enteric pathogens and can thereby promote good gastrointestinal health. PMID- 16042075 TI - The anticoagulant fraction from the leaves of Diospyros kaki L. has an antithrombotic activity. AB - The leaves of Persimmon (Diospyros kaki L.) has long been used for tea in Korea since it was thought to be effective against hypertension. An anticoagulant fraction was purified through gel filtration G-100, hydrophobic, gel filtration G 150, and FPLC, Phenyl superpose column chromatographies. The purified fraction was homogenous and its Mr was estimated 10,000 Da by gel filtration and SDS-PAGE. The purified fraction was sensitive to treatment of subtilisin B, but not to heat and its activity was not changed after periodate oxidation, indicating that the activity was not due to carbohydrates. It delayed thrombin time (TT), activated partial thromboplastin time (APTT), and prothrombin time (PT) using human plasma. TT was more sensitive than APTT and PT, suggesting that the anticoagulant activity may be caused by a degradation or a defect of fibrin or thrombin. It did not cause the hydrolysis of fibrin after incubation. However, it inhibited thrombin-catalyzed fibrin formation with a competitive inhibition pattern. These results indicate that it may be an antithrombotic agent and that it is bound to fibrinogen binding sites of thrombin. PMID- 16042076 TI - Inhibitory effects of furoquinoline alkaloids from Melicope confusa and Dictamnus albus against human phosphodiesterase 5 (hPDE5A) in vitro. AB - Eight furoquinoline alkaloids were purified from two plants belonging to the Rutaceae family. Kokusaginine, skimmianine, evolitrine, and confusameline were purified from Melicope confusa, and haplopine, robustine, dictamine, and gamma fagarine from Dictamnus albus. In this study, the eight furoquinoline alkaloids were examined for inhibitory potency against human phosphodiesterase 5 (hPDE5A) in vitro. DNA encoding the catalytic domain of human PDE5A was amplified from the mRNA of T24 cells by RT-PCR and was fused to GST in an expression vector. GST tagged PDE5A was then purified by glutathione affinity chromatography and used in inhibition assays. Of the eight alkaloids, gamma-fagarine was the most potent inhibitor of PDE5A, and its single methoxy group at the C-8 position was shown to be critical for inhibitory activity. These results clearly illustrate the relationship between PDE5A inhibition and the methoxy group position in furoquinoline alkaloids. PMID- 16042077 TI - Effects of Korean red ginseng extract on cisplatin-induced nausea and vomiting. AB - Ginseng, the root of Panax ginseng C.A. Meyer, is well known as a tonic medicine for restoring and enhancing human health. In traditional medicine, ginseng is utilized for the alleviation of emesis, which includes nausea and vomiting. However, it has not yet been demonstrated whether ginseng exhibits in vivo anti nausea and anti-vomiting properties. In this study, we examined the anti-emetic effect of Korean red ginseng total extract (KRGE) on cisplatin-induced nausea and vomiting using ferrets. Intraperitoneal administration (i.p.) of cisplatin (7.5 mg/kg) induced both nausea and vomiting with one-hour latency. The episodes of nausea and vomiting reached a peak after 1.5 h and persisted for 3 h. Treatment with KRGE via oral route significantly reduced the cisplatin-induced nausea and vomiting in a dose-dependent manner. The anti-emetic effect was 12.7 +/- 8.6, 31.8 +/- 6.9, and 67.6 +/- 4.0% with doses of 0.3, 1.0, and 3.0 g/kg of KRGE, respectively. Pretreatment with KRGE via oral route 1 and 2 h before cisplatin administration also significantly attenuated the cisplatin-induced nausea and vomiting. However this did not occur with a pretreatment 4 h before cisplatin administration. These results are supportive of KRGE being utilized as an anti emetic agent against nausea and vomiting caused by chemotherapy (i.e. cisplatin). PMID- 16042078 TI - G1 phase arrest of the cell cycle by a ginseng metabolite, compound K, in U937 human monocytic leukamia cells. AB - We recently reported that the ginseng saponin metabolite, compound K (20-O-beta-D glucopyranosyl-20(S)-protopanaxadiol, IH901), inhibits the growth of U937 cells through caspase-dependent apoptosis pathway. In this study, we further characterized the effects of compound K on U937 cells and found that, in addition to apoptosis, compound K induced the arrest of the G1 phase. The compound K treated U937 cells showed increased p21 expression; an inhibitory protein of cyclin-cdk complex. The up-regulation of p21 was followed by the inactivation of cyclin D and the cdk4 protein, which act at the early G1 phase, and cyclin E, which acts at the late G1 phase. Furthermore, compound K induced the activation of JNK and the transcription factor AP-1, which is a downstream target of JNK. These findings suggest that the up-regulation of p21 and activation of JNK in the compound K treated cells contribute to the arrest of the G1 phase. PMID- 16042079 TI - Improvement of memory by dieckol and phlorofucofuroeckol in ethanol-treated mice: possible involvement of the inhibition of acetylcholinesterase. AB - Phlorotannins, the polyphonic compounds found in brown Eisenia and Ecklonia algae, have several pharmacologically beneficial effects such as anti inflammation. In addition, our recent data show that these compounds may improve the cognitive functions of aged humans suggesting the potential ability to enhance memory in several neurodegenerative disorders. To examine the experimental hypothesis that two effective components of Ecklonia cava, dieckol and phlorofucofuroeckol (PFF), have memory-enhancing abilities, both were administered orally to mice before a passive avoidance test. The repeated administration of either dieckol or PFF dose-dependently reduced the inhibition of latency by the administration of ethanol. To investigate the mode of memory enhancing actions, the levels of major central neurotransmitters in three different regions (striatum, hippocampus, and frontal cortex) of the mouse brain were measured. The levels of some of the neurotransmitters were significantly changed by ethanol. Both dieckol and PFF altered the levels of some neurotransmitters modified by the ethanol treatment. It is noteworthy that both dieckol and PFF increased the level of acetylcholine, and they exerted anticholinesterase activities. Overall, the memory-enhancing abilities of dieckol and PFF may result from, at least in part, the increment of the brain level of acetylcholine by inhibiting acetylcholinesterase. PMID- 16042080 TI - Influence of naloxone on catecholamine release evoked by nicotinic receptor stimulation in the isolated rat adrenal gland. AB - The present study was designed to investigate the effect of naloxone, a well known opioid antagonist, on the secretion of catecholamines (CA) evoked by cholinergic stimulation and membrane-depolarization in the isolated perfused rat adrenal glands, and to establish its mechanism of action. Naloxone (10(-6) approximately 10(-5) M), perfused into an adrenal vein for 60 min, produced dose- and time-dependent inhibition of CA secretory responses evoked by ACh (5.32 x 10( 3) M), high K+ (5.6 x 10(-2) M), DMPP (10(-4) M) and McN-A-343 (10(-4) M). Naloxone itself also failed to affect the basal CA output. In adrenal glands loaded with naloxone (3 x 10(-6) M), the CA secretory responses evoked by Bay-K 8644, an activator of L-type Ca2+ channels, and cyclopiazonic acid, an inhibitor of cytoplasmic Ca(2+)-ATPase, were also inhibited. In the presence of met enkephalin (5 x 10(-6) M), a well known opioid agonist, the CA secretory responses evoked by ACh, high K+, DMPP, McN-A-343, Bay-K-8644 and cyclopiazonic acid were also significantly inhibited. Taken together, these results suggest that naloxone greatly inhibits the CA secretion evoked by stimulation of cholinergic (both nicotinic and muscarinic) receptors as well as that by membrane depolarization. It seems that these inhibitory effects of naloxone does not involve opioid receptors, but might be mediated by blocking both the calcium influx into the rat adrenal medullary chromaffin cells and the uptake of Ca2+ into the cytoplasmic calcium store, which are at least partly relevant to the direct interaction with the nicotinic receptor itself. PMID- 16042081 TI - Induces vasodilatation of rat mesenteric artery in vitro mainly by inhibiting receptor-mediated Ca(2+)-influx and Ca(2+)-release. AB - The purpose of this study was to investigate the effect of atropine on peripheral vasodilation and the mechanisms involved. The isometric tension of rat mesenteric artery rings was recorded in vitro on a myograph. The results showed that atropine, at concentrations greater than 1 microM, relaxed the noradrenalin (NA) precontracted rat mesenteric artery in a concentration-dependent manner. Atropine induced vasodilatation was mediated, in part, by an endothelium-dependent mechanism, to which endothelium-derived hyperpolarizing factor may contribute. Atropine was able to shift the NA-induced concentration-response curve to the right, in a non-parallel manner, suggesting the mechanism of atropine was not mediated via the (alpha1-adrenoreceptor. The beta-adrenoreceptor and ATP sensitive potassium channel, a voltage dependent calcium channel, were not involved in the vasodilatation. However, atropine inhibited the contraction derived from NA and CaCI2 in Ca(2+)-free medium, in a concentration dependent manner, indicating the vasodilatation was related to the inhibition of extracellular Ca2+ influx through the receptor-operated calcium channels and intracellular Ca2+ release from the Ca2+ store. Atropine had no effect on the caffeine-induced contraction in the artery segments, indicating the inhibition of intracellular Ca2+ release as a result of atropine most likely occurs via the IP3 pathway rather than the ryanodine receptors. Our results suggest that atropine induced vasodilatation is mainly from artery smooth muscle cells due to inhibition of the receptor-mediated Ca(2+)-influx and Ca(2+)-release, and partly from the endothelium mediated by EDHF. PMID- 16042082 TI - Role of neuropeptide Y and proopiomelanocortin in fluoxetine-induced anorexia. AB - Fluoxetine is an anorexic agent known to reduce food intake and weight gain. However, the molecular mechanism by which fluoxetine induces anorexia has not been well-established. We examined mRNA expression levels of neuropeptide Y (NPY) and proopiomelanocortin (POMC) in the brain regions of rats using RT-PCR and in situ hybridization techniques after 2 weeks of administering fluoxetine daily. Fluoxetine persistently suppressed food intake and weight gain during the experimental period. The pair-fed group confirmed that the reduction in body weight in the fluoxetine treated rats resulted primarily from decreased food intake. RT-PCR analyses showed that mRNA expression levels of both NPY and POMC were markedly reduced by fluoxetine treatment in all parts of the brain examined, including the hypothalamus. POMC mRNA in situ signals were significantly decreased, NPY levels tended to increase in the arcuate nucleus (ARC) of fluoxetine treated rats (compared to the vehicle controls). In the pair-fed group, NPY mRNA levels did not change, but the POMC levels decreased (compared with the vehicle controls). These results reveal that the chronic administration of fluoxetine decreases expression levels in both NPY and POMC in the brain, and suggests that fluoxetine-induced anorexia may not be mediated by changes in the ARC expression of either NPY or POMC. It is possible that a fluoxetine raised level of 5-HT play an inhibitory role in the orectic action caused by a reduced expression of ARC POMC (alpha-MSH). PMID- 16042083 TI - Evaluation of transferrin-polyethylenimine conjugate for targeted gene delivery. AB - With the aim to improve the specificity and to reduce the cytotoxicity of polyethylenimine (PEI), we have synthesized the conjugates of the branched PEI (25 kDa) with transferrin. The transferrin-PEI (TP) conjugates with five compositions were synthesized using periodate oxidation method and confirmed by FT-IR spectroscopy and gel permeation chromatography. The free amine contents of TP conjugates, which were able to condense and deliver DNA, increased as the amount of PEI increased. TP/DNA polyplexes were characterized by measuring gel electrophoresis, ethidium bromide fluorescence quenching, particle size and zeta potential of complexes. Complete complexation of the polyplexes was observed above the N/P ratio of 5 in TP/ DNA, and above 3 in PEI/DNA, respectively. The zeta potential of the complexes decreased as the amount of transferrin in TP conjugates increased. Transfection efficiency of TP conjugates was evaluated in HeLa cell and Jurkat cell systems. Among the five compositions of TP conjugates, TP-2 system mediated a higher beta-galactosidase gene expression than PEI system in Jurkat cell which was known to express elevated numbers of transferrin receptors. From the results of the cell viability based on MTT assay, TP conjugates showed lower cytotoxicity compared with the PEI system. We expect that the TP conjugate can be used efficiently as a nonviral gene delivery vector. PMID- 16042084 TI - Polymorphism of doxazosin mesylate. AB - Nine polymorphic modifications of doxazosin mesylate have been obtained by recrystallization in organic solvents under variable conditions. Different polymorphs of doxazosin mesylate were characterized by powder X-ray crystallography diffractometry (PXRD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TG). Transformation of Form 1 and Form 2 was not occurred in three relative humidities (0%, 51%, and 99%) at 20 +/- 0.5 for 30 days. PMID- 16042085 TI - Drug release from xyloglucan beads coated with Eudragit for oral drug delivery. AB - Xyloglucan (XG), which exhibits thermal sol to gel transition, non-toxicity, and low gelation concentration, is of interest in the development of sustained release carriers for drug delivery. Drug-loaded XG beads were prepared by extruding dropwise a dispersion of indomethacin in aqueous XG solution (2 wt.-%) through a syringe into corn oil. Enteric coating of XG bead was performed using Eudragit L 100 to improve the stability of XG bead in gastrointestinal (GI) track and to achieve gastroresistant drug release. Release behavior of indomethacin from XG beads in vitro was investigated as a function of loading content of drug, pH of release medium, and concentration of coating agent. Adhesive force of XG was also measured using the tensile test. Uniform-sized spherical beads with particle diameters ranging from 692 +/- 30 to 819 +/- 50 microm were obtained. The effect of drug content on the release of indomethacin from XG beads depended on the medium pH. Release of indomethacin from XG beads was retarded by coating with Eudragit and increased rapidly with the change in medium pH from 1.2 to 7.4. Adhesive force of XG was stronger than that of Carbopol 943 P, a well-known commercial mucoadhesive polymer, in wet state. Results indicate the enteric coated XG beads may be suitable as a carrier for oral drug delivery of irritant drug in the stomach. PMID- 16042086 TI - WVSMA tackles proposed medicaid cuts. PMID- 16042087 TI - Hard choices. PMID- 16042088 TI - Management of esophageal carcinoma. AB - Esophageal cancer is a devastating disease and among the most lethal malignancies worldwide. Despite advances in chemotherapy and radiation techniques, tumor stage at the time of presentation is the most important predictor of patient survival. Surgical resection offers the only curative treatment currently employed and transhiatal, transthoracic and en bloc esophagectomy are all standard surgical procedures used in the treatment of this disease. The Surgical Oncology Service of the West Virginia University Hospital favors the transhiatal approach since it offers lower morbidity and mortality without compromising survival when compared with transthoracic and en bloc esophagectomies. PMID- 16042089 TI - A rare case of massive gastrointestinal hemorrhage caused by Meckel's diverticulum in a 53-year-old man. AB - A 53-year-old man with a history of lung cancer mestastic to the brain and pelvis presented with hypotension and massive GI bleeding. He had no past history of gastrointestinal disease or bleeding. The patient was found to have an unusual focal source of bleeding--a large ulcerated Meckel's diverticulum with acute hemorrhage. Meckel's diverticulum is the etiology of most gastrointestinal bleeding episodes in the pediatric population; however, bleeding from a Meckel's diverticulum in an adult is rare and only three other cases have been reported. We present this case report and a literature review. PMID- 16042090 TI - Screening for erectile dysfunction in the primary care practice: results of a survey. AB - To evaluate the screening patterns of primary care physicians (PCP) with regards to erectile dysfunction, a study was performed using an Institution Review Board Approved SHIM (Sexual Health Inventory for Men) questionnaire of 131 male patients presenting to the Family Practice Clinic at West Virginia University School of Medicine in Morgantown. A total of 109 surveys were completed and the data were compiled and analyzed with descriptive statistics using SPSS version 10.0 software. Of these patients, over 92% were Caucasian. The average age bracket for the study population was 40-50, and the average number of risk factors for ED, not including age, was 1.65. Forty-one percent of patients had a SHIM score less than or equal to 21, indicative of an element of ED. A total of 84% had primary care physicians, 22% of patients with a PCP were screened for ED. Of those who were screened by their PCP over half were initiated by the patient. As a result of this study, we believe the SHIM instrument should be performed on patients with any identifiable risk factor since effective treatment of ED is available, and ED can be associated with occult cardiac disease. PMID- 16042091 TI - Fracture of the osteoporotic cervical spine from a low-level trauma. AB - Most cervical spine fractures are caused by high-level trauma or by malignancy. Compression-type fractures usually affect the lumbar and thoracic spine below the T8 level and rarely the cervical spine. In the elderly, however, cervical spine compression fractures can occur with relatively minor trauma and are associated with high morbidity and mortality. PMID- 16042092 TI - A diagnostic dilemma: case study of a 36-year-old female with alpha-1-antitrypsin deficiency. AB - Alpha-1-antitrypsin deficiency is a genetic disorder that presents as early-onset emphysema in its most severe form. A high index of clinical suspicion is needed in cases of lung or liver disease of unknown etiology or a suggestive history and physical examination. Low or absent serum levels of alpha-1-antitrypsin levels identify persons with the disease and phenotyping is the confirmatory test. The main goal of management is attempting to prevent or slowing the progression of damage to the lungs. Medical and surgical options for treatment include augmentation therapy that maintains protective levels of AAT in the lung and serum and lung transplantation may be necessary in severe cases. We present this case report of a patient with Alpha-1-antitrypsin deficiency in order to increase awareness of this condition since early diagnosis improves long-term prognosis and reduces the overall cost of therapy. PMID- 16042093 TI - Coalition shares recommendations to help West Virginians live healthier. PMID- 16042094 TI - Be understanding and proactive when discussing patients' questions about payments. PMID- 16042095 TI - Take precautions against mosquitos, dead birds to prevent West Nile, encephalitis viruses. PMID- 16042096 TI - Progress in measles control: Zambia, 1999-2004. PMID- 16042097 TI - Radiographic evaluation of aspirated metallic foil foreign bodies. AB - OBJECTIVES: Aspirated objects generally represent items accessible to children. When metallic candy wrapper aspiration is questioned, radiographic studies may aid diagnosis. An infant with repeated chest radiographs negative for a metallic foreign body was found to have a multi-layer metallic candy wrapper in the left main bronchus. The purpose of this study was to determine whether conventional and dual-energy radiographic techniques exclude the presence of aspirated metallic foil wrappers. METHODS: Single-layer and multi-layer metallic candy wrappers were radiographically studied with conventional and dual-energy radiographic techniques in 3 tissue models. RESULTS: No single-layer metallic samples were detectable with conventional or dual-energy radiography. The multilayer samples were not detectable at less than 8 layers (pulmonary tissue model) or 16 layers (mediastinal model) by either conventional or dual-energy radiography. CONCLUSIONS: Conventional and dual-energy chest radiographic techniques do not reliably exclude the presence of aspirated metallic foil wrappers. PMID- 16042098 TI - Laryngeal electromyography for adult unilateral vocal fold immobility: a survey of the American Broncho-Esophagological Association. AB - OBJECTIVES: Diagnostic and management strategies for adult unilateral vocal fold immobility (UVFI) vary among otolaryngologists. The aim of this study was to determine the current attitudes and practices regarding laryngeal electromyography (LEMG) for the management of adult UVFI within a cohort of subspecialty laryngologists. METHODS: A 19-item instrument focused on diagnosis and management of adult UVFI was mailed to active members (n = 249) of the American Broncho-Esophagological Association (ABEA). The subset of questions related to LEMG is reviewed in this report. Statistical analysis using a chi2 test was performed. RESULTS: The survey response rate was 34% (n = 84), with 8 surveys returned incomplete because of pediatric-limited practices. Of the respondents, 75% (n = 57) rely on LEMG for evaluation of UVFI, whereas 25% (n = 19) do not use LEMG. Of those who use LEMG, 54% place their own electrodes and 44% interpret the LEMG results themselves. Monopolar electrodes are used by 57% (n = 25), bipolar electrodes by 27% (n = 12), and hooked-wire electrodes by 17% (n = 7). Muscles evaluated by LEMG include the thyroarytenoid (100%), cricothyroid (94%), posterior cricoarytenoid (70%), lateral cricoarytenoid (43%), cricopharyngeus (27%), and interarytenoid (17%). The LEMG is performed in an unblinded fashion by most respondents (85%), and many (66%) feel a more accurate result is obtained when clinical information is known. There was no statistically significant difference in use of LEMG, placement of electrodes, and interpretation of LEMG according to percentage of laryngology practice. CONCLUSIONS: The survey results demonstrate congruence among ABEA members in the utility of LEMG in the management of adult UVFI. Some variability was noted in the methods by which LEMG is performed and interpreted. PMID- 16042099 TI - Regenerative medicine of the trachea: the first human case. AB - OBJECTIVES: The objective of the present study was to demonstrate regenerative medicine of the tracheal tissue by using an in situ tissue engineering technique for airway reconstruction. METHODS: Based on the previous successful experimental animal studies, the current regenerative technique was applied to repair of the trachea of a 78-year-old woman with thyroid cancer. A Marlex mesh tube covered by collagen sponge was used as a tissue scaffold. The operative intervention included right hemithyroidectomy, resection of the trachea, and tracheoplasty using the scaffold. The right half of three rings of the trachea was resected, and the scaffold material was sutured to the defect of the trachea. RESULTS: After 2 weeks, the mesh collagen structure of the artificial material could be seen with endoscopy in most of the implanted area. The artificial material was covered with epithelial growth after 2 months. Epithelialization continued to cover the artificial material completely for 2 years without any complications. CONCLUSIONS: The current regenerative technique avoided tracheotomy, a second operation, and deformity. Good epithelialization has been observed on the tracheal luminal surface without any complications for 2 years. Although long term observation is required, regenerative medicine of the tracheal tissue appears feasible for airway reconstruction. PMID- 16042100 TI - Variability of specific airway resistance in patients with laryngeal hemiplegia. AB - OBJECTIVES: This study was designed to analyze whether respiratory flows and specific airway resistance (sRaw) depend on the degree of breathiness and on the position of the paralyzed vocal fold in laryngeal hemiplegia. METHODS: We performed a prospective study involving 55 patients affected by laryngeal hemiplegia. RESULTS: The paralyzed fold was in an intermediate position in 18 cases and in a paramedian position in 37. Breathiness was estimated with the GRBAS scale, and the patients were divided into four groups: B0 (12 patients), B1 (14), B2 (16), and B3 (13). Spirometry was used to measure the flow-volume loop, and body plethysmography was used to measure the sRaw at increasing respiratory frequencies (30 +/- 5, 60 +/- 5, and 90 +/- 5 breaths per minute). The mean inspiratory flows (PIF, FIF50) were lower than predicted (<80%) in all four groups; there was no significant intergroup difference. In all four groups, the mean FEF5o/FIF50 ratio was >1, as is typical of variable extrathoracic obstruction. The mean sRaw values increased with respiratory frequency, and the increase was higher in group B3, although the values varied widely. The frequency dependent increase in the sRaw value was not significantly related to the degree of breathiness, nor to the position of the paralyzed fold. Furthermore, Spearman's coefficient did not reveal any correlation between the sRaw values and inspiratory flows, showing that plethysmography and spirometry explore different aspects of airway function. CONCLUSIONS: Respiratory flows and sRaw are not significantly influenced by either the degree of breathiness or the position of the paralyzed vocal fold. PMID- 16042101 TI - Combined endoscopic transmaxillary-transnasal approach to the pterygoid region, lateral sphenoid sinus, and retrobulbar orbit. AB - OBJECTIVES: The sublabial transmaxillary approach to the pterygoid region was a popular one during the Caldwell-Luc, pre-endoscopic era. It was the procedure of choice for management of lesions of the pterygopalatine space, for internal maxillary artery ligation, and for vidian neurectomy. With the introduction of endoscopic instrumentation and techniques, the Caldwell-Luc procedure is rarely performed today. Also, because vidian neurectomy is performed less frequently, and because internal maxillary artery ligation for severe epistaxis has been replaced with transnasal endoscopic sphenopalatine artery ligation, the sublabial transmaxillary route is rarely used. We have found that combining the use of endoscopes via the sublabial approach and the transnasal endoscopic approach is very helpful for management of extreme lateral lesions of the sphenoid sinus, as well as the pterygoid region and the posterior orbit. METHODS: The records of patients who underwent a combined endoscopic transmaxillary-transnasal approach between 1994 and 2002 were reviewed. Indications for the procedure included extreme lateral sphenoid and pterygoid encephalocele (3 patients), pterygoid mucocele (2 patients), orbital apex lesion (2 patients), and pterygopalatine tumor (2 patients). RESULTS: Nine patients underwent the above-mentioned procedure. Wide and comfortable exposure of the involved region was achieved in all cases. Compared with the transnasal approach, the working distance, working comfort, and maneuverability of instruments were significantly enhanced. There were no major complications related to the approach. Because the maxillary wall opening is very small, infraorbital hypoesthesia is very limited and of short duration. CONCLUSIONS: Combining the transmaxillary and transnasal approaches for endoscopic management of pterygoid, lateral sphenoid, and retrobulbar orbit lesions provides excellent exposure and avoids the limited working angle and surgical struggle that may be associated with the use of the transnasal approach alone. PMID- 16042102 TI - Accuracy of measurements on digital videostroboscopic images of the vocal folds. AB - OBJECTIVES: The reliability of objective measurements on digital laryngeal images was investigated. METHODS: The magnitude of the error of measurement of surface areas by visually tracing the outline was determined for three different areas: the area of the lesion and, during vibration, the glottal area on maximal opening of the vocal folds and the glottal area on maximal closing of the vocal folds. RESULTS: The errors in these areas were 10% to 30%. The results suggest that the error is mainly due to the uncertainty of the real outline of areas with a vague boundary. Correction for differences in magnification between two images (posttherapy and pretherapy) is of importance in about 25% to 65% of cases, depending on the area measured. CONCLUSIONS: Only when the magnification ratio is small (less than about +/- 10% from 1.0) may a correction not be necessary. PMID- 16042103 TI - Midmembranous vocal fold lamina propria proteoglycans across selected species. AB - OBJECTIVES: We examined the proteoglycan (PG) and associated sulfated glycosaminoglycan (GAG) content of the midmembranous vocal fold lamina propria (LP) of humans, dogs, pigs, and ferrets. METHODS: The LP PG levels were assessed indirectly by quantifying the associated sulfated GAGs, and immunohistochemical analyses of specific PGs and/or GAGs (PGs/GAGs) were conducted. RESULTS: Sulfated GAGs constituted approximately (average +/- SEM) 14.7 +/- 2.1 microg per milligram of tissue total protein in the human LP--similar to levels in canine, porcine, and ferret LPs (p > .05). Immunohistochemical analysis identified versican, chondroitin 4- and 6-sulfate, and heparan sulfate in the LP extracellular matrix--PGs/GAGs previously believed to be localized only intracellularly and in the basement membrane. Observations of PG/GAG staining patterns resulted in identification of microstructurally based subdivisions of canine, porcine, and ferret LPs. CONCLUSIONS: The sulfated GAG concentration in human LP was similar to that of dermis. In contrast to the interspecies similarity in LP sulfated GAG levels, immunohistochemical analysis indicated notable interspecies differences in specific PG/GAG distributions. Moreover, spatial variations in the presence of several PGs/GAGs were observed--variations that may be integral in maintaining normal LP physiology. Finally, the noted canine, porcine, and ferret LP subdivisions may yield insight into the adaptation of LP microstructure to the phonatory needs of each species. PMID- 16042104 TI - Use of interferon-alpha in recurrent respiratory papillomatosis: 20-year follow up. AB - OBJECTIVES: The aim of this study was analysis of the results of use of interferon-alpha (IFN-alpha) in patients with recurrent respiratory papillomatosis (RRP) and correlation of the results with human papillomavirus (HPV) type. METHODS: A multicenter prospective series (42 patients from 22 hospitals) yielded 20 years of follow-up of patients with RRP and HPV typing who were treated with IFN-alpha in doses of 3 MU/m2 3 times per week. RESULTS: During long-term follow-up (mean +/- SD, 172 +/- 36.8 months), the rate of event-free survival evaluated by Kaplan-Meier analysis was 42.8%, and the overall survival rate was 82.6%. The HPV typing revealed an association of HPV 11 with a more aggressive disease course (64% of HPV 11 patients versus 24% of HPV 6 patients), a lower incidence of long-term response to IFN-alpha therapy (14% of HPV 11 patients versus 64% of HPV 6 patients), and a higher incidence of malignant transformation and mortality during follow-up (36% and 24%, respectively, of HPV 11 patients versus 0% of HPV 6 patients). CONCLUSIONS: The obtained results revealed maximal effectiveness of IFN-alpha therapy in RRP patients with HPV 6 as compared with HPV 11. The association of HPV 11 with a worse long-term response to IFN-alpha therapy and a higher incidence of malignant transformation and mortality is clinically important and indicates the necessity of HPV typing in RRP patients after the first biopsy. PMID- 16042105 TI - Quality of life following small fenestra stapedotomy. AB - OBJECTIVES: A cross-sectional observational study was designed to evaluate patient satisfaction after stapedotomy. METHODS: Two validated questionnaires, the Psychosocial Impact of Assistive Devices Scale (PIADS) and the Hearing Handicap Inventory (HHI), were used to assess patients for whom audiometric data were available. RESULTS: The response rate was 79%. The PIADS score showed that 96% of patients reported an improvement in quality of life. The HHI score, however, showed that 32% of patients still experienced a marked degree of handicap after surgery. The HHI scores correlated positively with postoperative audiometric data, and the PIADS scores correlated positively with the degree of change in audiometric data. CONCLUSIONS: The results of this study support the role of small fenestra stapedotomy as a primary treatment for otosclerosis. The study also identifies a certain subpopulation of patients with residual handicap who may require further aural rehabilitation. PMID- 16042106 TI - Timing of glottic closure during swallowing: a combined electromyographic and endoscopic analysis. AB - OBJECTIVES: We performed a case series to enhance our understanding of the coupling between neuromuscular events and glottic closure. METHODS: We performed combined flexible video laryngoscopy and electromyography in 4 healthy human subjects. Hooked-wire electrodes were placed in the superior pharyngeal constrictor, longitudinal pharyngeal, cricopharyngeus, thyroarytenoid, genioglossus, suprahyoid, and posterior cricoarytenoid muscles. A flexible endoscope tip was positioned in the oropharyngeal-hypopharyngeal region. The subjects performed multiple trials each of 10-mL normal and super-supraglottic liquid swallows. RESULTS: Arytenoid movement consistently preceded full glottic closure and was associated with cessation of activity of the posterior cricoarytenoid muscle. In 89% of normal swallows, the glottis was partially open in the video frame before bolus passage. The maximum amount of thyroarytenoid electromyographic activity occurred during endoscopic whiteout. When subjects executed a super-supraglottic swallow, early thyroarytenoid activity coincided with arytenoid contact. CONCLUSIONS: The initial medialization of the arytenoids is due to a decrease in motor tone of the posterior cricoarytenoid muscle. Full glottic closure typically occurs late in the process of swallowing, with activation of the thyroarytenoid muscle. Shifting of arytenoid medialization and glottic closure earlier in the super-supraglottic swallow indicates that glottic closure is under significant voluntary control. PMID- 16042107 TI - Office-based laryngoscopic observations of recurrent laryngeal nerve paresis and paralysis. AB - OBJECTIVES: To evaluate the endoscopic criteria of recurrent laryngeal nerve disorders, we performed a retrospective evaluation of videolaryngoscopic recordings from 50 patients with recurrent laryngeal nerve disorders. METHODS: The videolaryngoscopic examination was performed with rigid and flexible endoscopes. The range of motion of three laryngeal structures was assessed: the vocal ligament, the vocal process, and the arytenoid "hump" (mainly the corniculate region). RESULTS: Comparison of movement of these three structures revealed discrepancies. In 16 of 45 patients (36%) rigid endoscopy showed movements of the arytenoid hump associated with absence of any mobility of the vocal process and vocal ligament. In 5 patients the extent of movement of the vocal process and vocal ligament was less than that of the arytenoid hump. Only in 24 of 45 cases were the ratings for the vocal process, vocal ligament, and arytenoid hump identical. The findings of fiberscopy were comparable. CONCLUSIONS: In assessing recurrent laryngeal nerve disorders via laryngoscopy, sole judgment of the arytenoid hump movement can mislead. Our interpretation suggests that visible movement of the mucosa covering the arytenoid and accessory cartilages is not always paralleled by movement of the arytenoid cartilage itself. It was shown that the best criterion to rely on in endoscopy is movement of the vocal process or the vocal ligament. PMID- 16042108 TI - Congenital bilateral vocal cord paralysis and the role of glycine. AB - OBJECTIVES: We sought to modify normal laryngeal constrictor (LC) motoneuron activity to induce a pattern of aberrant LC muscle function that may serve as a model of congenital bilateral vocal cord paralysis. METHODS: Single unit extracellular recordings of functionally identified LC motoneurons were made in anesthetized Sprague-Dawley rats, and the response to both intravenous and iontophoretic application of the glycine antagonist strychnine was studied. RESULTS: The postinspiratory firing pattern of LC motoneurons became inspiratory after intravenous injection of strychnine (4 of 5 rats), but no change was recorded in response to strychnine iontophoresis (7 of 8 rats). CONCLUSIONS: Blockade of glycinergic inhibitory neurotransmission by strychnine, acting above the level of the LC motoneuron, causes LC motoneurons to fire during inspiration rather than after inspiration. This observation suggests that impaired glycine neurotransmission may be an underlying mechanism that explains the clinical manifestations of congenital bilateral vocal cord paralysis. PMID- 16042109 TI - Influence of nutrients on biomass evolution in an upflow anaerobic sludge blanket reactor degrading sulfate-laden organics. AB - This paper describes the effect of the nutrients iron (Fe), nickel (Ni), zinc (Zn), cobalt (Co), and molybdenum (Mo) on biomass evolution in an upflow anaerobic sludge blanket (UASB) reactor metabolizing synthetic sulfate-laden organics at varying operating conditions during a period of 540 days. A bench scale model of a UASB reactor was operated at a temperature of 35 degrees C for a chemical oxygen demand-to-sulfate (COD/SO4(2-)) ratio of 8.59 to 2.0, a sulfate loading rate of 0.54 to 1.88 kg SO4(2-)/m3 x d, and an organic loading rate of 1.9 to 5.75 kg COD/m3 x d. Biomass was characterized in terms of total methanogenic activity, acetate-utilizing methanogenic activity, total sulfidogenic activity, acetate-utilizing sulfidogenic activity, and scanning electron microscopy (SEM). Nickel and cobalt limitation appears to affect the activity of hydrogen-utilizing methane-producing bacteria (HMPB) significantly without having an appreciable effect on the activity of acetate-utilizing methane producing bacteria (AMPB). Nickel and cobalt supplementation resulted in increased availability and, consequently, restoration of biomass activity and process performance. Iron limitation and sulfidogenic conditions resulted in the growth of low-density, hollow, fragile granules that washed out, causing process instability and performance deterioration. Iron and cobalt supplementation indicated significant stimulation of AMPB with slight inhibition of HMPB. Examination of biomass through SEM indicated a population shift with dominance of sarcina-type organisms and the formation of hollow granules. Granule disintegration was observed toward the end of the study. PMID- 16042110 TI - The effect of organic and hydraulic shock loads on the production of soluble microbial products in anaerobic digesters. AB - Anaerobic chemostats were used to investigate the effects of organic and hydraulic shock loads on the production of soluble microbial products (SMP). Production of SMP was found to increase during glucose spikes, reaching up 15% of the influent chemical oxygen demand. These SMP appear to be utilization associated products produced as a result of the temporarily high organic load, and chemical analysis and ultrafiltration experiments revealed that most of these compounds are difficult to identify and that the majority of them are present in the low molecular weight (MW) range. Production of SMP also increased when the hydraulic retention time was reduced from 15 to 3 days, and an increase in DNA concentration in the bulk solution suggested enhanced cell lysis. Although the cause of lysis was not clear, it is believed that most of the SMP produced under such conditions were biomass-associated products following cell death. While the majority of these compounds lay in the low MW range, as much as 35% were found to have MWs greater than 1 kDa. During the period when the anaerobic chemostat was fed no alkalinity and the pH remained lower than 6.5 for more than a week, a slightly higher production of SMP and a shift in the MW distribution towards the production of higher MW SMP was observed. PMID- 16042111 TI - Removal of azo dye from water by magnetite adsorption-Fenton oxidation. AB - The aim of this study is to highlight the possibility of using powder magnetite adsorption-Fenton oxidation as a method for removal of azo dye acid red B (ARB) from water. The adsorption properties of magnetite powder towards ARB were studied. The oxidation of adsorbed ARB and regeneration of magnetite adsorbent at the same time by Fenton reagent (hydrogen peroxide [H2O2] + iron (II) [Fe2+]) in another treatment unit with a smaller volume was also investigated. The efficiency of Fenton oxidation of ARB was compared for the reaction carried out in solution and on magnetite. The magnetic separation method was used to recover magnetite after adsorption or regeneration. The results indicated that the adsorption rate was fast. The capacity was strongly dependent on pH and inorganic anions, and pH 3.8 was optimal for the adsorption of ARB. The adsorption can be described well using the Langmuir model. The oxidation was more efficient for ARB adsorbed on magnetite than in solution. The adsorption capacity of magnetite increased significantly after regeneration, which was the result of an increase in surface area of the adsorbent and change of elemental ratio (oxygen:iron [O:Fe]) on the surface. The maximum adsorption capacity for ARB was 32.4 mg/g adsorbent. PMID- 16042112 TI - Study on the dynamics of algal bloom and its influence factors in Tolo Harbour, Hong Kong. AB - In this paper, the semi-enclosed bay named Tolo Harbour and Channel in Hong Kong, which was frequently attacked by red tides, was used as a case study. Data sets related to marine water quality, river nutrients, and meteorological conditions recorded between 1988 and 1999 were chosen for statistical analysis. A multivariate analysis showed that algal growth, represented by the chlorophyll a concentration, had obvious spatial and temporal variations in the study area. The chlorophyll a concentration had a consistently decreasing trend from the inner part of the Harbour and surface waters to the outer part and bottom waters. The temporal variations had a markedly seasonal variation with high bioproductivity in spring and winter. There were long-term fluctuations in the chlorophyll a concentration with a high-low-high pattern in the study period. Nutrients and hydrological and meteorological conditions were important factors of algal bloom. Besides nitrogen, which was the most critical factor of algal bloom for the whole water body, total phosphorus in the surface waters and phosphate (PO4) and silica (SiO2) in the bottom waters also showed strongly positive or negative correlations with the chlorophyll a level. For the meteorological conditions, global solar radiation was the key factor of massive algal bloom in the study period, while rainfall and wind direction were the most important factors of seasonal variation. PMID- 16042113 TI - The effect of pH, temperature, and molecular size on the removal of dyes from textile effluent using manganese oxides-modified diatomite. AB - This work is an investigation on the use of manganese oxides-modified diatomite (MOMD) for the removal of color from textile wastewaters. The modification of the diatomite was carried out by treatment with manganese oxides; delta-bimessite type resulted. The surface area, pH(ZPC), Fourier transform infrared (FTIR), and scanning electron microscopy of MOMD were studied. The influence of concentration, pH, particle size, and temperature on the adsorption capacities of methylene blue (MB) and hydrolyzed reactive black (RB) and reactive yellow (RY) was investigated. Key thermodynamic parameters such as deltaH degrees, deltaS degrees, and deltaG degrees were also determined. The deltaH degrees of MB and hydrolyzed RB and RY was +94.64, -143.1, and -38.78 kJ/mol, respectively, whereas the deltaG degrees values of -17.68, -27.93, and -23.26 kJ/mol were obtained, indicating a spontaneous process with low activation-energy requirements. The findings further indicated that coulomb interaction, molecular size, and orientation of the dye and its surface charge played an important role in the adsorption and attachment of the dyes to the bimessite layers of MOMD. PMID- 16042114 TI - Disinfection and dewatering of wastewater solids by interstitial vapor generation. AB - Disinfection of wastewater solids (waste activated solids [WAS]) by interstitial vapor generation was investigated. In addition to the magnitude of disinfection, the amount of water removed and cost relative to traditional residuals disinfection processes was also examined. The process of interstitial vapor generation occurs as a result of the rapid heating of liquid in the interstices of the solid-liquid array. Intense heating causes boiling of the slurry liquid, resulting in an expanding vapor front that simultaneously dewaters the wastewater solids and contributes to the destruction of viable pathogenic microorganisms. Objectives of the study were threefold: (1) to validate disinfection of WAS using the interstitial vapor technique; (2) establish the degree of possible drying of the residuals using the techniques; and (3) establish the key operating variables for the process. Results showed a significant reduction in the most probable number of total coliforms and Escherichia coli (E. coli). Specifically, greater than four-log unit reductions were produced for both total coliform and E. coli bacteria. In addition to quantifying the reduction in bacteria, the percent solids were increased from an initial amount of 7.6% (mass basis) to a final solids content greater than 90% using optimal processing conditions. Cost comparisons were also conducted and shown to be quite favorable when compared with traditional disinfection methods such as lime addition. Because of the high level of E. coli reduction achieved, the process of interstitial vapor generation is shown to be capable of converting a class B biosolids into a class A pathogen reduced product. For example, an initial most probable number (MPN) of 1.2 x 10(6) E. coli bacteria were reduced to 19 at the extreme conditions of the process, well below the requirement of an MPN of 1000 for fecal coliform bacteria. Given its ability to disinfect and dewater wastewater solids, the interstitial vapor generation process was found to be a robust and beneficial technique to produce an environmental and publicly acceptable recyclable biosolids resource. PMID- 16042115 TI - Statistical distributions of uncertainty and variability in activated sludge model parameters. AB - All models used in activated sludge design and analysis use parameters to characterize process performance. The values of these parameters are often assumed based on default values recommended in the literature, but to date, no quantitative estimates of the parameter uncertainties have been published. Similarly, little attention has been given to quantifying site-specific parameter variability, even though its occurrence has been observed several times in the literature. In this paper, universal uncertainty distributions of the model parameters from Activated Sludge Model No. 1 are developed from a database of parameter values reported in the literature using Bayesian statistics. Site specific distributions of parameter variability were developed using the same techniques. All parameter distributions developed demonstrated that significant uncertainty and variability exist, which could lead to overdesign or plant failure if not considered during the design process. PMID- 16042116 TI - A combination of ion exchange and electrochemical reduction for nitrate removal from drinking water. Part I: nitrate removal using a selective anion exchanger in the bicarbonate form with reuse of the regenerant solution. AB - The process of selective nitrate removal from drinking water by means of ion exchange was studied. A commercial strong base anion exchanger with triethylammonium (-N+Et3) functional groups was used in the bicarbonate (HCO3-) and carbonate (CO3(2-)) form. The aim of this study was to optimize ion-exchanger regeneration in view of the subsequent electrochemical reduction of nitrates in the spent regenerant solution. The effects of ion-exchanger form, concentration of regenerant solution, and presence of nitrates, chlorides, and sulphates in the regenerant solution were studied. The strong base anion exchanger in HCO3- form that was investigated was able to treat 270 bed volumes of model water solution containing 124 mg dm(-3) nitrates. To achieve adequate regeneration of the saturated anion exchanger, it is necessary to use approximately 30 bed volumes of fresh 1-M sodium bicarbonate (NaHCO3) regenerant solution. The presence of residual 50-mg dm(-3) nitrates in the regenerant solution, treated by electrolysis, resulted in an increase in the dose of regenerant solution to 35 bed volumes and a decrease in the subsequent sorption run of approximately 13%. The volume of applied regenerant solution was high, but the consumption of NaHCO3 for regeneration was low. PMID- 16042117 TI - A combination of ion exchange and electrochemical reduction for nitrate removal from drinking water. Part II: electrochemical treatment of a spent regenerant solution. AB - The process of electrochemical treatment of a solution after strong basic anion exchanger regeneration was studied. The goal of the study was to reduce the nitrate content in the solution to allow its use in a closed loop. Diaphragmless, flow-through cells in a recirculation mode with and without a fluidizing bed of inert particles in the interelectrode space equipped with copper (Cu) cathodes and activated titanium anodes were used. The temperature was maintained at 20 degrees C. To assess the influence of recirculation of the regenerant solution on the quality of the treated water, the effect of the addition of copper ions to the solution, postelectrolysis cathode treatment, and enhanced mass transfer on the electrolysis results with respect to current efficiency and residual nitrate and nitrite concentration were investigated using an artificial solution. On the basis of the experimental results, a laboratory-scale unit for selective nitrate removal was designed and constructed that integrated ion exchange and electrochemical cell to one assembly. The process of recirculation of regenerant solution was tested using groundwater. PMID- 16042118 TI - Effect of alum addition on the performance of submerged membranes for wastewater treatment. AB - The effect of aluminum sulfate (alum) addition on membrane performance was investigated, with a particular focus on membrane fouling. During initial operation, alum was added and the performance monitored. After terminating alum addition, the transmembrane pressure (TMP), which is indicative of membrane resistance to flow or fouling, increased. Accompanying the increase in TMP was an increase in the organic nonsettleable fraction (colloidal + dissolved) content of the mixed liquor and deterioration of permeate quality and floc strength. Permeate polysaccharide concentrations increased significantly, suggesting a preferential binding of solution polysaccharides by alum. Upon reinitiating alum addition, the TMP only partially recovered, indicating some irreversible fouling, while mixed liquor nonsettleable organic material, permeate quality, and floc strength returned to initial levels. These results suggest that direct alum addition to membrane bioreactors can improve membrane performance by reducing the organic fouling material and improving floc structure and strength. It appears that bulk liquid polysaccharides may contribute to irreversible membrane fouling, and this fraction can be efficiently controlled through the alum addition. PMID- 16042119 TI - Decolorization and degradation of dyes with mediated fenton reaction. AB - A mediated Fenton system has been evaluated for decolorization of several types of dyes. The result shows that the Fenton system with a dihydroxybenzoic acid (DHBA) chelator-mediator effectively reduced the color of a diluted solution of Carta Yellow RW liquid, Carta Yellow G liquid, and Cartasol Red 2GF liquid dye to a colorless level after 90 minutes of treatment with 100 microM iron II (Fe[II]), 100 microM DHBA, and 10 mM hydrogen peroxide (H2O2) at room temperature. Our results show that compared to a neat Fenton process, the mediated Fenton decolorization process increased the production, and therefore the effective longevity, of hydroxyl radical (OH) species to increase the decolorization efficiency. Our results suggest that application of this system would also result in an increase in dissolved oxygen (DO) in solution, which in turn would result in reduction of chemical oxygen demand (COD), biochemical oxygen demand (BOD), and total organic carbon (TOC). PMID- 16042120 TI - Of: Determining critical flow conditions for chloride impairment in an effluent dominated, storm-peaking, western U.S. stream, L. D. Duke; E. Erickson, 75, 39 (2003). PMID- 16042121 TI - Dr. Herbert H. Lindner. PMID- 16042122 TI - Evaluation of cyclodextrins modified with dichloro-, dimethyl-, and chloromethylphenylcarbamate groups as chiral stationary phases for capillary electrochromatography. AB - Capillary electrochromatography using cyclodextrins modified with dichloro-, dimethyl-, and chloromethylcarbamate groups were used for the enantiomeric separation of standard analytes. The chiral selector was chemically bonded to aminopropylsilanized silica particles, and these chiral stationary phases (CSPs), mixed with aminopropylsilanized silica (1:1 wt:wt), were packed into 100-microm i.d. fused-silica capillaries. The effect of the type of cyclodextrin, the nature and position of the substituents on the phenyl ring, and the binding mode of cyclodextrin phenylcarbamates onto the silica gel surface on the chiral recognition were studied. Experimental parameters such as organic solvent concentration were varied in order to better understand the mechanism contributing to the chiral recognition of these CSPs. Good enantioseparations were achieved for a racemic flavanone (FLA) and trans-cyclopropanedicarboxylic acid dianilide (CAD). PMID- 16042123 TI - Experimental design applied for the simultaneous analysis of whey proteins and caseins of binary and ternary milk mixtures by capillary electrophoresis. AB - A capillary electrophoresis method for the simultaneous separation of caseins, whey proteins, and para-kappa-casein that appears during the manufacture of cheese was optimized using the response surface methodology. The parameters selected for this study were pH, voltage, and temperature. Under the optimized conditions (30 kV at 20 degrees C with 10 mM phosphate buffer at pH 3) casein proteins alpha(s)-casein; beta-casein, including genetic variants A1, A2, and B, kappa-CN, and para-kappa-CN; and whey proteins alpha-lactalbumin and beta lactoglobulin (A and B variants) were separated. The method was applied successfully to skim milk, to casein precipitated at pH 4.6, and to a model sample containing rennet casein and milk. The milk used was of three types: cow, ewe, and goat. The present procedure can be easily applied to the separation of the major bovine, ovine, and caprine milk proteins in binary and ternary milk mixtures. PMID- 16042124 TI - Evaluation of the effect of somatic cell counts on casein proteolysis in ovine milk cheese by means of capillary electrophoresis. AB - The primary proteolysis of ewe's cheeses made from milk with different somatic cell counts (5CC) included three groups of study: cheeses made with milk with less than 500,000 somatic cells (SC); with milk containing 1 x 10(6) to 1.5 x 10(6) SC, the normal range for ewe's milk; and with milk containing more than 3.106 SC. These were examined by capillary eledrophoresis. Intact caseins as alpha(s1)-CN (I, II, and III), alpha(s2)-CN, and beta-CN (beta, beta1, and beta2) were identified. Some peptides resulting from casein proteolysis as gamma-CN and l-alpha(s1)-CN were also identified. The study reveals that proteolysis increased with ripening time and the extent of casein degradation depended on the content in SC, showing that residual intact casein, both alpha(s1)-CN and beta-CN, decreased as the SCC of milk increased because of their higher proteolytic enzyme levels. The proteolysis pattern indicated that several enzymes were implicated in increased proteolysis in high SCC cheeses. PMID- 16042125 TI - Improved CZE capabilities with new dynamic coatings. AB - Electroosmotic flow (EOF) is one of the fundamental processes affecting both resolution and separation times in capillary zone electrophoresis (CZE). EOF is a function of pH and buffer composition (zeta potential) in bare-silica capillaries at any pH above 3 and runs in the cathodal direction, decreasing the effective separation length for cationic species. On the other hand, the absence of EOF at low pH can significantly increase separation times, particularly for low pl zwitterionic species. This interdependence of pH and EOF limits the ability to design effective separations by capillary zone electrophoresis. The EOTrol family of dynamic coatings (Target Discovery, Inc., Palo Alto, CA, U.S.A.) allows both the magnitude and direction of EOF to be adjusted independently of the buffer composition and pH. Here we report the use of EOTrol in two challenging CZE separations: (1) inorganic cations with small mobility differences, and (2) the rapid separation of organic zwitterions that are only resolvable at low pH. PMID- 16042126 TI - Micellar electrokinetic capillary chromatography analysis of phthalic acid isomers. AB - A capillary electrophoretic method for the analysis and separation of ortho- and para-phthalic acid isomers was developed. The best separation was achieved using micellar electrokinetic capillary chromatography (MECC) performed in a 110 mM borate buffer pH 8.3 also containing 40 mM sodium dodecyl sulfate (SDS) and 20% methanol. The effect of the addition of methanol, acetonitrile, and beta cyclodextrin on the separation of the isomers was investigated. The buffer system developed was shown to provide a dependable method of analysis from which the concentration of each analyte could be reliably determined. PMID- 16042127 TI - Micropreparative separation, fractionation, and peptide mapping of beta lactoglobulin A and B variants by capillary electrophoresis. AB - The methodological aspects for the separation, fractionation, and peptide mapping by free zone capillary electrophoresis (CZE) of beta-lactoglobulin (beta-Lg) variants A and B were established. First, beta-Lg variants A or B were separated and fractionated by CZE. Then, the collected protein fraction was subjected to off-line tryptic digestion. Second, peptide mapping of the tryptic hydrolysates and peptide fraction collection were carried out by CZE. beta-Lg variants were separated and collected using an uncoated capillary (72 cm x 75 microm i.d.) in 0.05 M borate buffer containing 0.1% Tween 20 at pH 8.0 by applying 20 kV. By subjecting the capillary under pressure after a delay time of 15%, the protein was collected in a microvial containing digestion buffer. The most suitable conditions for the tryptic digestion of beta-Lg were established by monitoring the reaction products with CZE. A tryptic hydrolysis with an enzyme-to-substrate ratio (E/S) of 1/20 and incubation for 20 hr at 37 degrees C was found to result in the most suitable conditions. Peptides were separated and collected using an uncoated capillary (120 cm x 75 microm i.d.) in 0.15 M formic acid at pH 2.3 by applying 28 kV. Peptide maps were highly reproducible as shown by coefficients of variation of less than 0.89 and 5.42% for migration times and peak areas, respectively. Moreover, very good resolution of the peptide maps revealed the region in which the aberrant peptides of the beta-Lg variants may be located. PMID- 16042128 TI - [Neurologic disease due to adenovirus infection]. AB - The aim of this study was to assess the prevalence of adenovirusm (ADV) infections in neurological disorders. A total of 108 cerebrospinal fluid (CSF) samples from 79 encephalitis cases, 7 meningitis and 22 other neurological diseases analysed in our laboratory between 2000 and 2002 were studied. Forty nine (47.4%) belonged to immunocompromised patients. Viral genome was detected using nested polymerase chain reaction (Nested-PCR) and ADV genotypes were identified using partial gene sequence analysis of hexon gene. Adenovirus were detected in 6 of 108 (5.5%) CSF samples tested. All of these were from encephalitis cases, 6/79, representing 7.6% of them. No statistically significant differences were observed (p > 0.05) between the immunocompromised and non immunocompromised patients with ADV infection of the central nervous system. Two ADV genotypes (B1 and C) were identified. In conclusion, our results describe the role of ADV in neurologic infections in Argentina. The results contribute to the knowledge of ADV epidemiology, specially in encephalitis. PMID- 16042129 TI - [Group B streptococcus colonization during pregnancy and prevention of early onset of disease]. AB - Group B Streptococcus (GBS) is the most frequent cause of early onset of neonatal sepsis. Case-fatality rate is 6-20% for newborns. Neurological sequel occurs in 30% of survivors. In 1996, the Centers for Disease Control and Prevention (CDC), the American College of Obstetricians and Gynecologists, and the American Academy of Pediatrics recommended that obstetrics providers should adopt either a culture based or a risk-based approach for the prevention of this disease. The aim of this prospective study was to determine the colonization rate of GBS in our population of pregnant women between July 1st 2001 and December 31st 2002, and to introduce a culture-based strategy to prevent early onset neonatal GBS disease. From a population of 1756 pregnant women, 1228 were screened with rectal and vaginal swabs (69.9%). Maternal colonization rate was 1.4% (17 patients). There was one case of early-onset neonatal sepsis consistent with GBS disease (0.6%) in a patient with negative cultures. From the colonized patients, only one presented risk factors. Because most of the colonized women did not present intrapartum risk factors, the results of this study suggest that the culture-based approach should be used for the prevention of early-onset GBS disease in our population. Cost-benefit studies are needed in our country to determine if this prevention strategy is able to be implemented in all the settings of Argentina. PMID- 16042130 TI - [A focus study from a case of human fascioliasis in Neuquen]. AB - An epidemiological focal study was performed in Loncopue, Neuquen, Argentina, in November 2002 to detect the origin of the infection in a human case of fascioliasis confirmed by an indirect-ELISA test, six months before the study. Thirty five individual fecal samples were taken from domestic livestock, and watercress plants and snails were collected from the irrigation ditches connected to a main canal in the surroundings of the patient's house. A new blood sample was taken from the already recovered patient. The patient was still seropositive to Fasciola hepatica antigens. No metacercariae were found in the 222 watercress leaves checked. All the snails collected (n=130) were identified as Lymnaea viatrix and two out of 101 (2%) were infected with F. hepatica larvae. Coprological analysis showed F. hepatica eggs in 100% of goats (10/10), 82% of sheep (9/11) and 86% of bovines (6/7). The number of eggs per gram shed by positive goats (median=20.7, Q1=6.2, Q3=34.5) and sheep (4, 1.8, 13) was significantly higher than in cows (0.3, 0.3, 1.7) (p < 0.01). Local veterinary control programs were apparently not effective in this case. Anthelmintics used and treatment schedule should be revised and small herds raised at households should also be included and treated. PMID- 16042131 TI - Immunochemical and morphometric features of astrocyte reactivity vs. plaque location in Alzheimer's disease. AB - The quantitative relationship between glial fibrillary acidic protein (GFAP) hyper-reactivity and beta-amyloid protein (betaAP) deposition was investigated by double immunoperoxidase labeling of hippocampal and entorhinal cortex sections from five Alzheimer's disease (AD) cases and five age-matched controls. betaAP plaques, which were absent in controls, were found in all AD samples, without significant differences in number or perimeter according to their location among the regions studied. In contrast, the mean number of GFAP (+) cells was significantly greater in the hippocampus than in the entorhinal cortex from AD cases (49 vs.39). Although at lower values (30 vs. 20), predominance of astrocyte hyperplasia in hippocampus as compared with entorhinal cortex was also found in control samples. Concomitant astrocyte hypertrophy, as defined by surface density (Sv) values of GFAP-immunoreactive material exceeding those of control means, affected a similar proportion of cells in the hippocampus (73%) and the entorhinal cortex (74%) from AD cases. Since an increased number of GFAP (+) cells in the hippocampus was not accompanied by an increased number and/or perimeter of neighbouring plaques, such differential hyper-reactivity in samples from AD patients, as well as in those with normal aging, seems to depend partially on the regional location of the involved astrocyte. PMID- 16042132 TI - [Immunoglobulin heavy chain gene rearrangements in the monoclonal gammopathies]. AB - Plasma cell neoplasia occurs as a result of the expansion of an immunoglobulin secreting B-cells clones, known as monoclonal component or M component. Malignant neoplasias include multiple myeloma and Waldenstrom macroglobulinemia, while premalignant conditions comprise monoclonal gammopathies of unknown significance (MGUS). MGUS present a monoclonal component with no signs of multiple myeloma, Waldenstrom macroglobulinemia, primary amyloidosis or other disorders. Pathological, radiological and clinical features are required for the diagnosis. Approximately 25% of patients with MGUS will become multiple myeloma, primary amiloidosis, macroglobulinemia, or other lymphoproliferative disease, which would be a premyelomatous condition. The objective of this study was to determine the clinical implications of immunophenotyping by flow cytometry and of the detection of clonality by molecular biology. A total of 32 patients were studied. Seven of them were diagnosed with multiple myeloma, and 25 with monoclonal gammopathy under study. These 32 patients were divided into four groups, based on their clinical data and flow cytometry outcome. In patients with non-diagnostic flow cytometry detection of immunoglobulin heavy chain gene rearrangements by PCR was performed, and monoclonality was found in 59% of the cases. The study of immunoglobulin heavy chain gene rearrangements by molecular biology allows a more sensitive detection of clonality. PMID- 16042133 TI - [Toxocariasis in children from a subtropical region]. AB - Toxocariasis is present worldwide but people living in areas with sanitary deficiencies are considered at the highest risk of infection, particularly children. The aim of this work was to know clinical and immunological aspects of infantile toxocariasis in a subtropical region in Argentina. For this purpose, 182 children of both sexes, 0-16 years old and with eosinophilia higher than 10%, living in Resistencia City (Northeast Argentina) were studied. Clinical examination, personal and epidemiological data recording, parasitological fecal examination and dosage of Toxocara canis IgG and IgM levels by EIE were performed; all positive sera were confirmed by Western Blot. Out of 182 children, 122 were positive for T. canis-IgG (67.0%); 28.8% lack of potable drinking water at home, 58.8% lack of sewerage facilities, 91.1% referred a close contact with dogs or cats, 30.0% had a history of geophagia and 86.7% lived along streets without pavement. The clinical forms of the infection were: 77.8% asymptomatic, 6.7% ocular larva migrans and 15.5% visceral larva migrans. In 22 children the serological follow up until 18 months after treatment showed 10 children without change in IgG levels, in 11 it decreased but remained high, and in one it increased. There were 19 children with high levels of T. canis-IgM; in 8 it decreased along treatment, in one it remained stable and 10 of them became negative. There was one child considered as a re-infection case. Results highlight the importance that health authorities should assign to this infection in regions with sanitary deficiencies, where toxocariasis is usually not recognized as a relevant public health problem. PMID- 16042134 TI - [Chronic urticaria with alterations of the thyroid function and thyroid peroxidase antibodies]. AB - Chronic urticaria is a frequent pathology, characterized by the presence of hives and/or angioedema lasting longer than 6 weeks. In an important number of patients it behaves as an autoimmune illness, frequently associated with alterations in thyroid function and thyroid antibodies. We herein describe a consecutive series of 70 patients with a diagnosis of chronic urticaria. Seven (10%) had a diagnosis of thyroid illness previous to their first consultation. Thyroid function and thyroid antibodies were studied in the remaining 63 patients by measuring the level of serum thyrotropin and the titer of peroxidase antibodies. Abnormal thyrotropin levels were detected in 11 (17%) patients, who in conjunction with the 7 patients with previous thyroid illness, add up to 18 (26%) with altered thyroid function. From 61 patients who were tested for thyroid peroxidase antibodies, 22 (36%) were positive. Of 57 patients without a diagnosis of previous thyroid disease, in whom both the levels of serum thyrotropin and the presence of thyroid peroxidase antibodies had been studied, 24 (42%) presented at least one altered study. Given the high percentage of thyroid alterations in our series of patients, it seems clinically relevant to study the thyroid function and the presence of thyroid peroxidase antibodies in patients with chronic urticaria. PMID- 16042135 TI - [HLA DQB1 genotyping in latent autoimmune diabetes of adults (LADA)]. AB - Autoimmune diabetes is a complex, multifactorial disease caused by the interaction of genetic and environmental factors. This autoimmune diabetes is commonly manifested in childhood and adolescence with a fast onset (type 1 diabetes, IDDM) and it can occur in adult patients with a slow onset with delayed insulin requirement, (latent autoimmune diabetes in adults, LADA ). Autoimmune diabetes has strong class II HLA association mainly with DQB gene which constitutes the first susceptibility locus. However, association with the 5'INS- VNTR and CTLA-4 genes has been established. In this study, we analysed the polimorphic allele frequencies of DQB HLA gene in 63 LADA patients, 70 IDDM and 79 control subjects. The HLA DQB1 alleles typing was detected through Olerup SSP DQ kit using sequence specific primers. We observed a positive association of *0201-*0302 and *0201-*0201 genotypes in both types of diabetic patients compared to the control group (p < 0.05). Moreover, *0201-*0302 genotype was higher in IDDM than in LADA (p < 0.05). On the other hand, the *0602 protective allele analysis showed a high prevalence in the normal group compared to the diabetic population. In Argentina, the most frequent allele of susceptibility in LADA and IDDM patients was the *0201. Summing up, the finding of an increase in the *0201 allele, both in allelic and genotypic frequencies, allows the characterisation of our population of patients, LADA and IDDM, unlike other populations, in which the most frequent allele is *0302. PMID- 16042136 TI - [Lepidopterism due to the butterfly Hylesia nigricans. Preventive research intervention in Buenos Aires]. AB - A preventive research-intervention against the butterfly Hylesia nigricans was performed after an increase in spontaneous reports of nonspecific dermatitis in El Pato neighbourhood, Berazategui County, Buenos Aires Province. The overall pre intervention incidence was estimated with a semistructured questionnaire carried out during March 2001. The trees in the street were sprayed with Bacillus thuringiensis, and the insecticide effect was evaluated both in the field and in the laboratory (November 2002). The impact was estimated with a post-intervention questionnaire during March 2002. The age distribution, clinical appearance, persistence (11 days) and seasonality (January-February 2001) of the cases were consistent with dermatitis due to H. nigricans. The mortality of larvae 96 h after the spraying was 100%. The incidence rates before and after the intervention were 10.3% y 1.8% respectively. The coordinated action of National, Provincial and District oficials allowed to identify the problem, to design an operational research, and to perform a preventive control strategy transferable to the community. PMID- 16042137 TI - [The attitude of physicians regarding the promotion strategies of the pharmaceutical industry]. AB - Pharmaceutical companies invest large sums of money promoting their products. They use a multifaceted approach to drug promotion, incorporating techniques such as hospital and office detailing by pharmaceutical representatives. Although these practices are commonly used, little has been published about the attitude of physicians concerning their interaction with the pharmaceutical industry. We performed a cross sectional anonymous survey to identify the extent of and attitudes towards the relationship between the physicians and the pharmaceutical industry and its representatives with its impact on the knowledge, attitude and behavior of the physicians. Internists, cardiologists and dermatologists who work in ambulatory settings from private and public hospitals in Buenos Aires city participated in this study, 44% were female, 35% residents, 65% staff physicians, averaging 41 years of age. Of these, 86% receive medical samples frecuently, 39% desk gifts, 19% invitations to congresses and 12% free lunches. Half of the doctors believe that receiving benefits from the pharmaceutical industry has an influence on medical prescription, but only 27% accept this as influential in their own prescriptions. Residents consider, more frequently than others, that these activities affect their decisions, (42% vs. 18% p = 0.007, global 30%). Most of the participants consider appropriate receiving these benefits, although 35% think that they affect the final price of medications. In conclusion, there is a high level of interaction between the pharmaceutical industry and our medical population. Although the latter recognize the influence of these interactions on prescriptions and the elevation of the cost of the final product, they find it appropriate to receive benefits. PMID- 16042138 TI - [Platypnea-orthodeoxia syndrome, atrial septal aneurysm and right hemidiaphragmatic elevation with a right-to-left shunt through a patent foramen ovale]. AB - Platypnea-orthodeoxia is an uncommon syndrome of dyspnea and hypoxemia induced by upright position, which is subsequently relieved by recumbency. The case reported involved a 75-year-old man with a seven days history of dyspnea in the upright position. The chest radiograph and CT scan demonstrated an elevated right hemidiaphragm. A surface echocardiogram with saline solution showed a patent foramen ovale (PFO) and atrial septal aneurysm. A pulmonary arteriogram was performed showing no evidence of pulmonary embolism and normal pulmonary arterial pressures. Later, he also developed dyspnea in the supine position and a transesophageal echocardiogram with contrast demonstrated a large right to left shunt through a PFO. The closure by a catheter-deployed double-umbrella device caused an immediate improvement in the patient's oxygenation. PMID- 16042139 TI - [Calcified chronic pancreatitis in an HIV positive patient]. PMID- 16042140 TI - [Dextrocardia and auricular fibrillation]. PMID- 16042141 TI - [The thyroid as a model for molecular mechanisms in genetic diseases]. AB - Thyroid diseases constitute a heterogeneous collection of abnormalities associated with mutations in genes responsible for the development of thyroid: thyroid transcription factor-1 (TTF-1), thyroid transcriptions factor-2 (TTF-2) and PAX8, or in one of the genes coding for the proteins involved in thyroid hormone biosynthesis such as thyroglobulin (TG), thyroperoxidase (TPO), hydrogen peroxide-generating system (DUOX2), sodium/iodide symporter (NIS), pendrin (PDS), TSH and TSH receptor (TSHr). Congenital hypothyroidism occurs with a prevalence of 1 in 4000 newborns. Patients with this syndrome can be divided into two groups: nongoitrous (dysem/bryogenesis) or goitrous (dyshormonogenesis) congenital hypothyroidism. The dysembryogenesis group, which accounts for 85% of the cases, results from ectopy, agenesis and hypoplasia. In a minority of these patients, the congenital hypothyroidism is associated with mutations in TTF-1, TTF-2, PAX-8, TSH or TSHr genes. The presence of congenital goiter (15% of the cases) has been linked to mutations in the NIS, TG, TPO, DUOX2 or PDS genes. The congenital hypothyroidism with dyshormonogenesis is transmitted as an autosomal recessive trait. Somatic mutations of the TSHr have been identified in hyperfunctioning thyroid adenomas. Another established thyroid disease is the resistance to thyroid hormone (RTH). It is a syndrome of reduced tissue responsiveness to hormonal action caused by mutations located in the thyroid hormone receptor beta (TRbeta) gene. Mutant TRbetas interfere with the function of the wild-type receptor by a dominant negative mechanism. In conclusion, the identification of mutations in the thyroid expression genes has provided important insights into structure-function relationships. The thyroid constitutes an excellent model for the molecular study of genetic diseases. PMID- 16042142 TI - [Platypnea-orthodeoxia syndrome]. AB - Platypnea-orthodeoxia is an uncommon syndrome of dyspnea and hypoxemia induced by upright posture, which is subsequently relieved by recumbency. Traditionally, this condition has been reported in association with pulmonary, hepatic and cardiac diseases, but the mechanism is different in each situation. In presence of an atrial septal defect, a right to left cardiac shunt resulting as a consequence of redirection of the inferior vena cava flow towards the atrial septum and results in postural hypoxemia. In pulmonary shunts, as in hepatopulmonary syndrome and a-v pulmonary fistulas, the mechanism of hypoxemia is related to the preferential circulation to basal areas of both lungs in the upright position. On the other hand, lung diseases affecting basal areas and increasing the alveolar pressure are related to the development of parenchimal pulmonary shunts with hypoxemia during postural changes, since the dead space, the functional shunt and the hypoxic vascular pulmonary constriction are affected. The autonomic neuropathy may attenuate normal sympathetic vasoconstrictor responses during postural changes, resulting in ventilation perfusion mismatching and hypoxemia. Clinical and therapeutic aspects of this sindrome are provided. PMID- 16042143 TI - [Infectious diseases in classical Greece. Disease and contagion]. PMID- 16042144 TI - [Watercress and other dangerous foods]. PMID- 16042145 TI - [Aquiles Roncoroni (1923-2005) was also a public health physician]. PMID- 16042146 TI - [Cyclooxygenase-2 inhibitors and the ethics of research]. PMID- 16042147 TI - [Santiago Ramon y Cajal. An anniversary]. PMID- 16042148 TI - A method for the solvent extraction of low-boiling-point plant volatiles. AB - A new method has been developed for the extraction of volatiles from plant materials and tested on seedling tissue and mature leaves of Arabidopsis thaliana, pine needles and commercial mixtures of plant volatiles. Volatiles were extracted with n-pentane and then subjected to quick distillation at a moderate temperature. Under these conditions, compounds such as pigments, waxes and non volatile compounds remained undistilled, while short-chain volatile compounds were distilled into a receiving flask using a high-efficiency condenser. Removal of the n-pentane and concentration of the volatiles in the receiving flask was carried out using a Vigreux column condenser prior to GC-MS. The method is ideal for the rapid extraction of low-boiling-point volatiles from small amounts of plant material, such as is required when conducting metabolic profiling or defining biological properties of volatile components from large numbers of mutant lines. PMID- 16042149 TI - Chemical identification of the sources of commercial Fructus Chebulae. AB - Twenty-eight commercial samples of Fructus Chebulae were collected from local herbal markets in Taiwan and were determined to have been derived from Terminalia chebula Retz. and Terminalia chebula Retz. var. parviflora Thwaites, which differ markedly in external appearance. Ten tannin-related constituents [gallic acid (1), chebulic acid (2), punicalagin (4), chebulanin (7), corilagin (8), neochebulinic acid (9), ellagic acid (11), chebulagic acid (12), chebulinic acid (13) and 1,2,3,4,6-penta-O-galloyl-beta-D-glucose (14)] were identified and quantified by HPLC. Samples derived from T. chebula. var. parviflora, which are typically round-shaped, generally contained higher concentrations of 13 and 14 but lower levels of 12 compared with those from T. chebula, which are largely oval-shaped. The ratio of the concentration of 14 to that of 4 may serve as a potential parameter for differentiating samples from the two origins (T. chebula, ratio 0.6 +/- 0.3; T. chebula. var. parviflora, ratio 3.4 +/- 2.2). Levels of the three major bioactive constituents 12-14 were found to provide good references for the quality assessment of Fructus Chebulae. The ratio of the concentration of 12 to that of 14 may offer a guideline for determining quality as well as origin of the drug (lower-grade T. chebula, ratio 12.4 +/- 6.0; medium-grade T. chebula, ratio 8.8 +/- 7.9; higher-grade T. chebula, ratio 3.2 +/- 0.8; T. chebula var. parviflora, ratio 1.6 +/- 0.7). PMID- 16042150 TI - Determination of diarylheptanoids from Alpinia officinarum (Lesser Galangal) by HPLC with photodiode array and electrochemical detection. AB - Normal-phase column chromatography followed by semi-preparative reversed-phase HPLC has been used to isolate, from the rhizomes of Alpinia officinarum, five diarylheptanoids identified as 5-hydroxy-7-(4"-hydroxy-3"-methoxyphenyl)-1-phenyl 3-heptanone, 5-methoxy-7-(4"-hydroxy-3"-methoxyphenyl)-1-phenyl-3-heptanone, 7 (4"-hydroxyphenyl)-1-phenylhept-4-en-3-one, 7-(4"-hydroxy-3"-methoxyphenyl)-1 phenyl-hept-4-en-3-one, 1,7-diphenylhept-4-en-3-one. The levels of these five diarylheptanoids in root material were determined quantitatively by HPLC with UV detection and the assay methods so developed were simple, rapid and accurate. Four of the diarylheptanoids could also be detected by HPLC with electrochemical detection (ECD) in the oxidative mode, and ECD was found to have a higher sensitivity than photodiode array detection. PMID- 16042151 TI - Determination of quinolizidine alkaloids in Sophora tonkinensis by HPCE. AB - A simple, rapid and reliable high-performance capillary electrophoresis method has been developed to determine quantitatively the alkaloid content of Sophora tonkinensis, a Chinese herb commonly known as shan-dou-gen. A total of seven quinolizidine alkaloids (cytisine, sophocarpine, matrine, lehmannine, sophoranol, oxymatrine and oxysophocarpine) could be readily separated within 15 min. The running buffer was 50 mM phosphate buffer (pH 2.5) containing 1% hydroxypropyl beta-cyclodextrin and 3.3% isopropanol in water. The applied voltage was 25 kV, the capillary temperature was 25 degrees C, the detection wavelength was 200 nm and scopolamine butylbromide was used as internal standard. The method was used to analyse the chemical constituents of two commercial alternatives to shan-dou gen. The alkaloid constituents of authentic shan-dou-gen gave a specific HPCE electropherogram that could be used to distinguish the drug from potential substitutes. Furthermore, the content of oxymatrine and the total content of the seven quinolizidine alkaloids could be used as quantitative markers in order to assess the quality of S. tonkinensis. PMID- 16042152 TI - A phytochemical study of the quinolizidine alkaloids from Genista tenera by gas chromatography-mass spectrometry. AB - Gas chromatography coupled with mass spectrometry has been used to analyse the alkaloids present in the aerial parts of Genista tenera. Anagyrine, cytisine, N formylcytisine, N-methylcytisine and lupanine were the major compounds, the last two alkaloids being known for their hypoglycaemic activity. Dehydrocytisine, 5,6 dehydrolupanine, rhombifoline, aphylline and thermopsine were the minor alkaloids. The characterisation of the constituents was based on comparison of their Kovats retention indexes and electron impact-mass spectrometric data recorded on-line with those of reference compounds and literature data. PMID- 16042153 TI - Development and validation of a method for the quantitative determination of aflatoxin contaminants in Maytenus ilicifolia by HPLC with fluorescence detection. AB - A method for the quantification of aflatoxins B1, G1, B2 and G2 in the medicinal herb Maytenus ilicifolia was developed and validated. The method used immunoaffinity columns for sample clean-up and HPLC with fluorescence detection without any derivatisation step. The method showed good inter-day accuracy (bias values in the range 4.5-10.7%) and precision (5-16% RSD) when applied to the determination of levels of aflatoxins ranging from 7 to 20 ppb in the plant material. The detection limits for samples of the plant material spiked with aflatoxins were 3.5 ng/g for B1 and G1 and 0.1 ng/g for B2 and G2. The method was successfully applied to commercial samples of Maytenus ilicifolia for the screening of aflatoxin contaminants. PMID- 16042154 TI - Determination of major ginsenosides in Panax quinquefolius (American ginseng) using high-performance liquid chromatography. AB - In order to determine the active ingredients in root extracts of Panax quinquefolius (American ginseng), a gradient HPLC method involving UV photodiode array detection was applied to separate and quantify simultaneously the ginsenosides Rb1, Rb2, Rc, Rd, Re, Rf and Rg1. All ginseng saponins were baseline resolved under the selected conditions, and the detection limits were 1.0 microg/mL or less. The method has been applied to analyse ginsenosides extracted from American ginseng cultivated in both Wisconsin and Illinois. Ginsenosides Re and Rb1 were the two main ginseng saponins in the root. The amounts of Re in 5- and 7-year Illinois-cultivated samples were greater than those found in ginseng cultivated for 3 or 4 years in Wisconsin, whereas the levels of Rb1 were greater in the younger Wisconsin samples. PMID- 16042155 TI - Isolation of secondary metabolites from roots of Plumbago auriculata Lam. by countercurrent chromatography. AB - Centrifugal liquid-liquid partition chromatography presents significant advantages for the separation and purification of plant metabolites owing to the short operational time of the process and the elimination of possible irreversible adsorption of compounds. The crude chloroform extract from roots of Plumbago auriculata was analysed by countercurrent chromatography using hexane:ethyl acetate:methanol:water (40:10:10:2, v/v) as solvent system. The isolation of the naphthoquinones plumbagin and epi-isoshinanolone, the steroids sitosterol and 3-O-glucosylsitosterol, plumbagic and palmitic acids was easily achieved. Naphthoquinones are typical components of Plumbago species and they show interesting biological activities. PMID- 16042156 TI - Circadian rhythm of anti-fungal prenylated chromene in leaves of Piper aduncum. AB - Leaves of Piper aduncum accumulate the anti-fungal chromenes methyl 2,2-dimethyl 2H-1-chromene-6-carboxylate (1) and methyl 2,2-dimethyl-8-(3'-methyl-2'-butenyl) 2H-1-chromene-6-carboxylate (2). The enzymatic formation of 2 from dimethylallyl diphosphate and 1 was investigated using cell-free extracts of the title plant. An HPLC assay for the prenylation reaction was developed and the enzyme activity measured in the protein extracts. The prenyltransferase that catalyses the transfer of the dimethylallyl group to C-2' of 1 was soluble and required dimethylallyl diphosphate as the prenyl donor. In the leaves, the biosynthesis of the prenylated chromene 2 was time-regulated and prenyltransferase activity depended upon circadian variation. Preliminary characterisation and purification experiments on the prenyltransferase from P. aduncum have been performed. PMID- 16042157 TI - Current awareness in phytochemical analysis. PMID- 16042158 TI - Determinants of healthy eating in Canada: an overview and synthesis. AB - This article uses a population health perspective to examine the complex set of interactions among the determinants of healthy eating. An overview of current knowledge on determinants of healthy eating was organized as follows: 1) individual determinants of personal food choices and 2) collective determinants, including a) environmental determinants as the context for eating behaviours and b) public policies as creating supportive environments for healthy eating. A conceptual synthesis of the literature revealed that individual determinants of personal food choice (physiological state, food preferences, nutritional knowledge, perceptions of healthy eating and psychological factors) are necessary, but not sufficient, to explain eating behaviour, which is highly contextual. Collective determinants of eating behaviour include a wide range of contextual factors, such as the interpersonal environment created by family and peers, the physical environment, which determines food availability and accessibility, the economic environment, in which food is a commodity to be marketed for profit, and the social environment, in which social status (income, education and gender) and cultural milieu are determinants of healthy eating that may be working "invisibly" to structure food choice. Policy is a powerful means of mediating multiple environments. There are gaps in our understanding of the process of intervening in macro-level environments and the impact of such interventions on the promotion of healthy eating. Collective determinants of food choice and policy contexts for promoting healthy eating, therefore, require investment in research. Applying a population health promotion lens to understanding the multiple contexts influencing healthy eating provides insight into prioritizing research and action strategies for the promotion of healthy eating. PMID- 16042159 TI - Perceptions of healthy eating: state of knowledge and research gaps. AB - To effectively promote and support healthy eating among Canadians, there needs to be a better understanding of the factors that influence eating behaviours. Perceptions of healthy eating can be considered as one of the many factors influencing people's eating habits. For this review, "perceptions of healthy eating" are defined as the public's and health professionals' meanings, understandings, views, attitudes and beliefs about healthy eating, eating for health, and healthy foods. This article's aim is to review and summarize the literature on the perceptions of healthy eating and to identify the current state of knowledge and key knowledge gaps. Databases, the worldwide web, selected journals and reference lists were searched for relevant papers from the last 20 years. Reviewed articles suggest relative homogeneity in the perceptions of healthy eating despite the studies being conducted in different countries and involving different age groups, sexes and socio-economic status. Perceptions of healthy eating were generally based on food choice. Fruits and vegetables were consistently recognized as part of healthy eating. Characteristics of food such as naturalness, and fat, sugar and salt contents were also important in people's perceptions of healthy eating. Concepts related to healthy eating, such as balance, variety and moderation, were often mentioned, but they were found to be polysemous, conveying multiple meanings. The main gap identified in this review concerns the lack of knowledge available on perceptions of healthy eating. More data are needed on the perceptions of healthy eating in general, on the influence on perceptions of messages from diverse sources such as food companies, and, most important, on the role of perceptions of healthy eating as a determinant of food choice. PMID- 16042160 TI - Determinants of healthy eating in children and youth. AB - This review outlines the state of knowledge and research gaps in the area of determinants of healthy eating among children and youth. The article is structured around individual and collective determinants that affect healthy eating in children and youth. We defined healthy eating as "eating practices and behaviours that are consistent with improving, maintaining and/or enhancing health." Relevant databases were searched for papers published between January 1992 and March 2003 that focussed on children or youth and reported at least one factor relevant to healthy eating. Among collective factors, familial factors and the nature of foods available in the physical environment, including at home, schools and in fast-food establishments, stand out as significant influences on healthy eating in children and youth. The media, particularly television, also have an enormous potential influence and can overshadow familial influences. Individual factors identified include knowledge, attitudes and food preferences; only the latter have been identified as a strong determinant of healthy eating in both children and adolescents. The results of the review identified a significant body of literature in the area of determinants of healthy eating in children and youth; however, very little of this research has taken place in Canada. Only a few determinants, such as economic factors and food security, the content of media nutritional messages, and the issue of flavours, neophobia and food preferences, have undergone some examination by Canadian researchers. Research priorities for Canada in the area of determinants of healthy eating and surveillance of eating behaviours are identified. PMID- 16042161 TI - Determinants of healthy eating in community-dwelling elderly people. AB - Among seniors, food choice and related activities are affected by health status, biological changes wrought by aging and functional abilities, which are mediated in the larger arena by familial, social and economic factors. Determinants of healthy eating stem from individual and collective factors. Individual components include age, sex, education, physiological and health issues, psychological attributes, lifestyle practices, and knowledge, attitudes, beliefs and behaviours, in addition to other universal dietary determinants such as income, social status and culture. Collective determinants of healthy eating, such as accessible food labels, an appropriate food shopping environment, the marketing of the "healthy eating" message, adequate social support and provision of effective, community-based meal delivery services have the potential to mediate dietary habits and thus foster healthy eating. However, there is a startling paucity of research in this area, and this is particularly so in Canada. Using search and inclusion criteria and key search strings to guide the research, this article outlines the state of knowledge and research gaps in the area of determinants of healthy eating among Canadian seniors. In conclusion, dietary self-management persists in well, independent seniors without financial constraints, whatever their living arrangements, whereas nutritional risk is high among those in poor health and lacking in resources. Further study is necessary to clarify contributors to healthy eating in order to permit the development and evaluation of programs and services designed to encourage and facilitate healthy eating in older Canadians. PMID- 16042162 TI - Determinants of healthy eating in Aboriginal peoples in Canada: the current state of knowledge and research gaps. AB - Aboriginal peoples are the original inhabitants of Canada. These many diverse peoples have distinct languages, cultures, religious beliefs and political systems. The current dietary practices of Aboriginal peoples pose significant health risks. Interventions to improve the nutritional status of Aboriginal peoples must reflect the realities of how people make food choices and therefore should be informed by an understanding of contemporary patterns of food procurement, preparation and distribution. Most of the literature documenting the health of Aboriginal peoples is primarily epidemiologic, and there is limited discussion of the determinants that contribute to health status. The majority of studies examining dietary intake in Aboriginal communities do not aim to study the determinants of food intake per se even though many describe differences in food intake across sex, age groups, seasons and sometimes communities, and may describe factors that could have an effect on food consumption (e.g., employment status, level of education, household size, presence of a hunter/trapper/fisher, occupation, main source of income). For these reasons, there are many gaps in knowledge pertaining to the determinants of healthy eating in Aboriginal peoples that must be filled. Given the diversity of Aboriginal peoples, research to address the gaps should take place at both the national level and at a more local level. Research would be important for each of Inuit, Metis and First Nations. PMID- 16042163 TI - Determinants of healthy eating among low-income Canadians. AB - This paper draws on four bodies of literature to consider the determinants of healthy eating for low-income Canadians: a) the social determinants of health; b) socio-economic gradients in diet; c) food security; and d) the sociology of food. Though there is a paucity of data for Canada, it is very likely that, as in other industrialized countries, there are socio-economic gradients in diet such that those who are better off consume healthier diets than those less well-to-do. The available evidence suggests that income affects food intake both directly and indirectly through the dispositions associated with particular social class locations. Thus, there may be both economic and cultural thresholds for some food groups or particular foods in food groups. Understanding these thresholds is especially important in addressing the issues facing those who are the most vulnerable among Canadians with low incomes: the food insecure. The literature reviewed suggests that improved nutrition for low-income Canadians may be difficult to achieve a) in isolation from other changes to improve their lives; b) without improvement in the nutrition of the general population of Canadians; and c) without some combination of these two changes. Four major areas of research need were identified: a) national data on socio-economic gradients in diet; b) sociological research on the interaction of income and class with other factors affecting food practices; c) sociological research on Canadian food norms and cultures; and d) research on the costs of healthy eating. PMID- 16042164 TI - Mental health and eating behaviours: a bi-directional relation. AB - BACKGROUND: Variations in mental health may contribute to or impair healthy eating. The relation between eating and mental health is bi-directional: one's mood or psychological state can affect what and how much one eats, and eating affects one's mood and psychological well-being. Thus, if we want to promote and develop strategies to encourage healthy eating, it is important to understand the connections between mental health and healthy eating. METHODS: To contribute to this understanding, we examine the research on individual differences in how people respond to food, as well as mood, and emotional, social and collective influences on what and how much is eaten; we then examine the implications of these connections for mental health, with a focus on adolescents and adults. Looking at the relation between eating and mental health from the other direction, we review research investigating whether the amount that one eats or particular foods one ingests can make one feel good or bad about oneself. CONCLUSIONS: Overeating and undereating have complex effects, sometimes contributing to improved feelings of well-being and at other times leaving the individual feeling guilty, deprived, depressed and anxious. We attempt to identify both what we know and the gaps in our knowledge. PMID- 16042165 TI - [Boost effect of recombinant IL-4 on protection of Schistosoma japonicum cathepsin B DNA vaccine in mice against the parasite]. AB - OBJECTIVE: To investigate the enhancement effect of IL-4 expression plasmid on cathepsin B DNA vaccine of Schistosoma japonicum (Sj) in mice. METHODS: The recombinant IL-4 plasmid constructed by cloning PCR amplified product of murine IL-4 gene into eukaryotic expression vector pcDNA3.1 was co-injected intramuscularly with Sj cathepsin B expression plasmid DNA to mice as the test group. The other three groups of mice were set up as control including IL-4 expression plasmid, Sj cathepsin B expression plasmid and two vacant vector plasmids. The expression of IL-4 and cathepsin B was visualized by immunohistochemistry. Challenge infection in mice was carried out 3 weeks after the last vaccination and immune protection was assessed by worm and egg reduction rates. RESULTS: The recombinant mIL-4 plasmid and cathepsin B DNA vaccine were expressed in muscular cells of the vaccinated mice. Immunization with cathepsin B DNA plus recombinant mIL-4 plasmid yielded a 43.2 % of worm reduction rate and a 76.6% of egg reduction rate, showing a significant difference (P<0.01, P<0.05) compared with that of cathepsin B DNA vaccine alone. CONCLUSION: As an adjuvant, IL-4 DNA can improve the protective effect of cathepsin B DNA vaccine in mice against S. japonicum infection. PMID- 16042166 TI - [Discovery of a new species of the pentastomid genus Porocephalus (Humboldt, 1811) from Taiwan, China and its pathogenic features]. AB - OBJECTIVE: To describe the morphological characteristics of Porocephalus taiwana sp. nov., discuss its pathogenic features and the method of etiological diagnosis of the new disease. METHODS: Fecal sedimentation concentration was used to collect nymphs from the patient's watery stool for species identification. Clinical information was collected for determining the pathogenic features of the new infection. RESULTS: A new pathogenic pentastomid Porocephalus taiwana sp. nov. is discovered and a new disease, porocephaliasis taiwana, is nominated. With the findings from this case it is proposed that the traditional visceral pentastomiasis should be divided into two subtypes, Encystic and Excystic. According to the pathological features, this case belongs to the excystic visceral pentastomiasis. CONCLUSION: Porocephalus taiwana sp. nov. is a new pathogenic pentastomid infecting humans. Porocephaliais taiwana belongs to a novel type (excystic) of visceral pentastomiasis. PMID- 16042167 TI - [Preliminary study on congenital transmission of Trichinella spiralis in mice]. AB - OBJECTIVE: To study congenital transmission of Trichinella spiralis in mice and observe the protection of anti-Trichinella antibodies from the infected dams to challenge infection. METHODS: According to the gestation (fertilization), the Kunming mice were divided into two groups: the infected group after gestation and the gestated group after infection. New-born mice were cut into small pieces to separate the larvae within 1 day after birth. One-day-old offspring born to normal dams were nursed by the infected dams, slaughtered after 21 days and examined for the larvae. Serum anti-Trichinella antibody level in offspring born to the infected dams was assayed by ELISA at different time after birth, and its immune protection against challenge infection was studied. RESULTS: Out of 6 offspring born to the dams infected at 7 days after fertilization, two were found to be infected. Among other female mice which were first infected with T. spiralis and then gestated, only the offspring born to the dams fertilized at 8 and 22 days after infection were found to be infected, the infection rate of offspring was 20% (2/10) and 25%(2/8) respectively. All larvae recovered from the young were non-encapsulated. The cross-fostering experiment showed that none of 30 offspring born to normal dams were found to be infected. The serum antibody positive rate in 27 offspring born to the infected dams at 1,7,24, and 40 days after birth was 100%, 100%, 77.8% and 14.8%, respectively. The worm reduction rate in the offspring 40 days after birth was 62.0% after challenge infection. The worm reduction rate in mice in which sera from the offspring born to the infected dams were passively transferred was 55.7%, there was a significant difference (P<0.01) compared to the control group. CONCLUSION: A transplacental transmission of T. spiralis is revealed in mice. Anti-Trichinella antibodies from the infected dams may partially protect the young from challenge infection. PMID- 16042168 TI - [Parasitism of Phthirus pubis on eyelashes]. PMID- 16042169 TI - [Comparison of PCR and isoenzyme analysis in identification of Anopheles minimus A and C]. AB - OBJECTIVE: To compare the PCR and isoenzyme methods in identification of Anopheles minimus A and C. METHODS: Samples of An. minimus collected in the field were identified from An. aconitus and An. jeyporiensis by PCR-RFLP after morphological examination, then classified by PCR-ASA. The mosquitoes identified by this method were further analyzed for isoenzyme to observe the difference, which was previously used as a technique to classify An. minimus A and C. RESULTS: PCR method can differentiate An. minimus A and C simply and quickly. For Isoenzyme method, only EST allozyme pattern displayed difference. CONCLUSION: PCR method is better than isoenzyme analysis in differentiating An. minimus A and C. PMID- 16042170 TI - [Gordiacea infection in urinary tract]. PMID- 16042171 TI - [Protective immunity induced by multivalent DNA vaccine of Schistosoma japonicum Mr23 x 10(3) membrane antigen and IL-12 in mice]. AB - OBJECTIVE: To develop multivalent DNA vaccine PV-IL12-Sj23 which co-expresses Sj23 and cytokine IL-12, and investigate its protective efficacy in BALB/c mice against challenge infection. METHODS: On the basis of the reconstructed plasmid PV-IL12-Sj23 and plasmid PV-IL12, blank plasmid PV and plasmid PV-Sj23 only expressing Sj23 were constructed. Fifty BALB/c male mice were divided into five groups, which were immunized intramuscularly with multivalent DNA vaccine PV-IL12 Sj23, plasmid PV-Sj23 expressing Sj23, plasmid PV-IL12 expressing cytokine IL-12, blank plasmid PV and saline, respectively. Each mouse was immunized with 100 microg DNA only once. All the mice were challenged with 40 cercariae at week 4, killed and perfused for collection of worms at week10. The number of recovered worms and eggs in the liver were counted. RESULTS: Blank plasmid PV and plasmid PV-Sj23 expressing Sj23 were successfully constructed. The worm reduction rate in PV-IL12-Sj23 group and PV-Sj23 group was 45.5% and 27.2% (P< 0.05) respectively. The number of eggs in liver tissue was reduced by 58.4% and 33.9% respectively. CONCLUSION: Multivalent DNA vaccine PV-IL12-Sj23 can induce protective immunity against Schistosoma japonicum in BALB/c mice significantly, with a better protective efficacy than the monovalent DNA vaccine PV-Sj23. PMID- 16042172 TI - [Acquisition and analysis of 149 ESTs and 18 novel genes of Schistosoma japonicum]. AB - OBJECTIVE: To acquire and analyze adult stage Schistosoma japonicum (Chinese strain) expressed sequence tags and new genes from an adult S. japonicum cDNA library, and to search new vaccine candidates and drug targets. METHODS: A cDNA library was constructed from adult stage S. japonicum. Clones were selected randomly from the cDNA library and were sequenced. ESTs and new genes were acquired after analysis in GenBank databases by BLAST and other programs. All ESTs and new genes were submitted to GenBank and received accession numbers. RESULTS: 149 ESTs were acquired from a total 382 clones that were randomly selected from the adult S. japonicum cDNA library. All ESTs were successfully submitted to the dbEST at Genbank. Some of them were homologous with sequences of male, female, egg, schistosomula, cercaria and miracidia of S. japonicum. 18 new genes of adult S. japonicum were acquired. Some genes were housekeeping genes and some genes might be interesting as vaccine candidates or drugs targets. CONCLUSIONS: The EST strategy is a rapid, efficient and economical method to acquire ESTs and to discover new genes of adult stage S. japonicum from cDNA libraries. PMID- 16042173 TI - [Myiasis in gastro-intestinal tract]. PMID- 16042174 TI - [Field trial on rapid detection of echinococcosis by dot immunogold filtration assay (DIGFA) with whole blood sample]. AB - OBJECTIVE: To establish a rapid, simple and reliable assay with samples of whole blood for diagnosis and epidemiological study on hydatidosis. METHODS: The dot immunogold filtration assay kit was developed and potato agglutinin was applied to blot blood quickly. RESULTS: Among 1 678 persons from prevalent area, the positive rate of DIGFA was 8.469 while that of image examination was 3.04%. Both DIGFA and image technique showed positive results in 43 cases. 8 cases with positive image but negative DIGFA were followed up for 16 months, which turned out that 3 cases with necrotic hydatid cysts, 2 cases with calcified hydatid cysts and 2 cases with benign hepatic cysts. 99 cases with positive DIGFA but negative image were also followed up for 16 months, 3 pulmonary hydatid cases were confirmed. Among 38 cases proved by operation and histopathology, the positive rate of DIGFA was 89.5%. 52 samples from non-prevalent area all showed negative DIGFA. Another 40 non-hydatidosis cases (10 samples of hepatic hemangioma, 10 of non-parasitic cysts of liver, 10 of primary hepatic carcinoma, 6 of pulmonary tuberculosis, 4 of lung cancer) also showed negative DIGFA. 190 samples were selected randomly and detected blindly by DIGFA with whole blood, DIGFA with serum and ELISA with serum to evaluate their diagnostic effect with no statistical difference (P>0.05). CONCLUSION: The DIGFA kit is rapid, simple and reliable in epidemiological study of hydatid disease, with an advantage of using whole blood sample instead of serum. PMID- 16042175 TI - [DNA/MVA combined immunization: antibody response to Plasmodium falciparum merozoite surface protein 1 in mice]. AB - OBJECTIVE: To explore the effect of DNA/MVA combined immunization in enhancing antibody response to MSP1. METHODS: DNA vaccine and recombined MVA were constructed based on synthesized MSP1 gene (3D7). BALB/c mice were primed with DNA solely or together with GM-CSF expressing plasmid and then boosted with rMVA/ 190. Serum IgG and subtype IgG1 and IgG2a were assayed by ELISA. All mice were challenged with allelic replaced Plasmodium berghei. RESULTS: Antibodies to MSP1 190 were detected after DNA immunization with an end-point dilution titer of 1:2500. When GM-CSF plasmid was added, the antibody end-point dilution titer reached 1:11150, with an increase of 53 and 10 times respectively after MVA boosting. Among them anti-19000 antibodies were prominent, 1/4-1/3 of total IgG in serum. However, when the mice were challenged with Pb-PfM19 no prolonged survival was observed (P>0.05). CONCLUSION: High titer antibodies can be elicited in mice by using codon optimized MSPI gene and DNA/MVA combined immunization. The specificity and protection of these antibodies is being further investigated. PMID- 16042176 TI - [Protective immunity of the recombinant Schistosoma japonicum specific very low density lipoprotein binding protein as a vaccine candidate]. AB - OBJECTIVE: To investigate the protective immunity against Schistosoma japonicum in mice immunized with recombinant specific very low density lipoprotein binding protein (SVLBP) and its potential as vaccine candidate. METHODS: Recombinant SVLBP antigen was over-expressed under IPTG induction and purified by Ni-NTA affinity chromatography. C57BL/6 mice were immunized three times with purified reSVLBP complexed with Freund's adjuvant, at biweekly intervals. Then 35+/-1 cercariae of S. japonicum were given to each mouse by abdominal skin 10 days after the 3rd immunization. 45 days later, all mice were sacrificed to collect adult worms and count liver eggs. serum samples were collected before immunization and after challenge respectively, and were probed the antigen specific antibodies using a panel of ELISAs. RESULTS: The worm burden and the egg deposition in liver tissue were reduced by 33.4% and 47.6% respectively in the immunized group, in comparison with the adjuvant control group (P<0.05). Higher titer (>1:6 400) of total IgG was observed after challenge infection. The vaccinated mice developed significantly higher levels of IgG2a, IgG2b, IgG1 than those of control mice. CONCLUSION: The recombinant tegumental SVLBP antigen could induce partial protection against S. japonicum infection. These data demonstrate the potential of SVLBP as a schistosome vaccine candidate. PMID- 16042177 TI - [Sequence analysis and expression of GRA7 gene of Toxoplasma gondii isolates in Escherichia coli]. AB - OBJECTIVE: To analyze the difference of GRA7 gene of Toxoplasma gondii different isolated strains and express GRA7 in Escherichia coli. METHODS: The GRA7 gene was amplified from genomes of T. gondii isolates by PCR and was cloned into pGEX-4T 1. The recombinant plasmid was transformed into JM109 and sent to be sequenced. The sequence was analyzed with CLUSTALW (an internet tool). The recombinant plasmid was induced by IPTG to express the fusion protein,which was identified by SDS-PAGE and Western blot with positive sera. The protein was purified and used as a diagnostic antigen for ELISA to test serum samples. RESULTS: There was no difference among the sequences of T. gondii GRA7 gene from different isolates. The recombinant plasmid pGEX-4T-1/GRA7 induced by IPTG was expressed in E. coli. It was a GST fusion protein and could react with human and rabbit positive sera analyzed by Western blot. CONCLUSION: The GRA7 gene of T. gondii isolates is highly conservative. The GRA7 is expressed as a recombinant protein in Escherichia coli, which shows an immunoreactivity. PMID- 16042178 TI - [Survival of the third stage larvae of anisakis simplex in various conditions and experimental infection in rats]. AB - OBJECTIVE To observe the characteristics of biology and pathogenicity of the third stage Anisakis simplex larvae (L3). METHODS: The survival time of the L3 in various conditions was observed and the pathological change after experimental infection in rats was examined. RESULTS: The results showed that the L3 frozen at -20 degrees C for 10 - 12 h can be killed. In the temperature range of 4 - 10 degrees C, the L3, can survive for over 8 months. The L3 was very active at 37 degrees C, and was killed at the high temperature over 40 degrees C in a very short time. The ingredients for sashimi cannot kill the L3. The experiment of rats infected by the L3 showed that about 15% -25% of the L3 penetrated into the gastrointestinal wall or migrated into the peritoneal cavity in 2 days. After 3d the L4 was not infectious, and died automatically in 7 - 10 days and could not develop into adults. The animals can be easily infected when the stomach was empty. The pathological study showed that the primary infection was a kind of reaction to foreign body, while that of the re - infection was allergic. CONCLUSION: The L3 has a strong resistance to low temperature and to ingredients, it can be killed by freezing at -20 degrees C in 24 hours. The L3 can not mature in the body of terrestrial mammals but causes pathological change in the stomach and allergy. PMID- 16042179 TI - [AIDS complicated by Clonorchis sinensis and Eimeria sp. infections]. PMID- 16042180 TI - [Development and identification of monoclonal antibodies against the recombinant P38 antigen of Schistosoma japonicum]. AB - OBJECTIVE: To express P38 of Schistosoma japonicum in Escherichia coli BL21 and develop the monoclonal antibodies (McAb) against rSjP38. METHODS: The recombinant plasmid pET32(a)-P38 was transformed into E. coli BL21 (DE3). 1 mmol/L IPTG (isopropyl-beta D-thiogalactopyranoside) was used to induce the expression of the recombinant rSjP38. The rSjP38 was soluble in supernatant after sonication and further purified by His-Ni chromatography. BALB/c mice were immunized with the purified rSjP38 and hybridomas were generated with traditional technique. McAbs were screened by ELISA with limited dilution. The subtype and specificity of McAb were identified by kit and Western blot respectively. RESULTS: Eight hybridoma cell lines secreting monoclonal antibodies to rSjP38 were obtained. The subtype of all the 8 McAbs are IgG1. Western blotting showed that the 8 McAbs reacted strongly and specifically with native antigen (P38) of Schistosoma japonicum. CONCLUSIONS: Eight hybridoma cell lines secreting highly specific McAbs against P38 have been established. PMID- 16042181 TI - [Survey and treatment of pulmonary acariasis among the workers involving in traditional Chinese medicinal materials]. AB - OBJECTIVE: To investigate the prevalence of pulmonary acariasis among the employees working on traditional Chinese medicinal materials and observe the effect of treatment. METHODS: History inquiry, detection of mites in sputum, blood examination for eosinophils and specific antibodies, x-ray chest film were carried out for 327 workers involving in traditional Chinese medicinal materials. Mites were found in sputum in 121 persons who were then treated with metronidazole, twice a day with a daily dosage of 0.8g for seven days as a course of treatment. Two courses were conducted with an interval of 7-10 day. Prevalence and morbidity in different groups of occupation, age, and sex were analyzed. RESULTS: The overall infection rate of mites in sputum was 37.0% (121/327) with an average morbidity of 12.5% (41/327). Among the four types of worker investigated, the highest infection rate (51.8%), and morbidity (18.6%) were in those working in transfer warehouse; the second highest infection rate (40.7%) and morbidity (15.7%) were in employees in factory of Chinese traditional medicine. Both groups showed a significant difference with others (chi2inf=11.36, P< 0.01; chi2inc=11.36, P<0.01). Higher morbidity was found in the middle-aged ones and those with more years of service. No difference was found between males and females (chi2=0.31, P>0.05). After treatment with metronidazole, 88.4% showed negative in sputum examination for mites and the efficacy of the treatment for pulmonary acariasis was 92.3%. CONCLUSIONS: Employees engaged in traditional Chinese medicinal materials are one of the groups at the highest risk of pulmonary acariasis. Metronidazole is effective in treating the infection. PMID- 16042182 TI - [Molecular markers related to antimalarial resistance of Plasmodium falciparum and their applications on the surveillance to drug resistance]. PMID- 16042183 TI - [The choice and analysis of spatial variables in the study of environmental factors of schistosomiasis]. PMID- 16042184 TI - [Observation on hepatic pathology in pigs experimentally infected by Taenia asiatica]. PMID- 16042185 TI - [Sampling survey on the risk factors of soil-transmitted helminth infection in Chongqing]. PMID- 16042186 TI - [Two cases of dermatitis induced by Ornithonyssus bacoti]. PMID- 16042187 TI - [Current status of human parasitic infections and the epidemiological factors in Kunming area]. PMID- 16042188 TI - The two faces of Rex Whistler (1905-1944). PMID- 16042189 TI - From dichoptic to dichotic: historical contrasts between binocular vision and binaural hearing. AB - Phenomena involving vision with two eyes have been commented upon for several thousand years whereas those concerned with hearing with two ears have a much more recent history. Studies of binocular vision and binaural hearing are contrasted with respect to the singleness of the percept, experimental manipulations of dichoptic and dichotic stimuli, eye and ear dominance, spatial localisation, and the instruments used to stimulate the paired organs. One of the principal phenomena that led to studies of dichotic hearing was dichoptic colour mixing. There was similar disagreement regarding whether colours or sounds could be combined when presented to different paired organs. Direction and distance in visual localisation were analysed before those for auditory localisation, partly due to difficulties in controlling the stimuli. Instruments for investigating binocular vision, like the stereoscope and pseudoscope, were invented before those for binaural hearing, like the stethophone and pseudophone. PMID- 16042190 TI - Contrast configuration influences grouping in apparent motion. AB - We investigated whether the same principles that influence grouping in static displays also influence grouping in apparent motion. Using the Ternus display, we found that the proportion of group motion reports was influenced by changes in contrast configuration. Subjects made judgments of completion of these same configurations in a static display. Generally, contrast configurations that induced a high proportion of group motion responses were judged as more 'complete' in static displays. Using a stereo display, we then tested whether stereo information and T-junction information were critical for this increase in group motion. Perceived grouping was consistently higher for same contrast polarity configurations than for opposite contrast polarity configurations, regardless of the presence of stereo information or explicit T-junctions. Thus, while grouping in static and moving displays showed a similar dependence on contrast configuration, motion grouping showed little dependence on stereo or T junction information. PMID- 16042191 TI - Asymmetric mislocalization of a visual flash ahead of and behind a moving objectt. AB - When subjects localize a flash relative to another stationary stimulus, the flash appears displaced in the direction of nearby motion signals (position capture; Whitney and Cavanagh, 2000 Nature Neuroscience 3 954-959). Our previous study had suggested that the position capture is larger for a flash presented ahead of a moving stimulus than for a flash behind it (Watanabe et al, 2003 Perception 32 545-559). In the present study, I investigated the spatial asymmetry of position capture. Experiment 1 demonstrated that asymmetric position capture occurs primarily in a moving-object-centered coordinate. Experiment 2 showed evidence that the asymmetric position capture operates after individuation of single visual objects. Finally, experiment 3 demonstrated that, when attention was reduced with a dual-task procedure, the asymmetric position capture increased. These results suggest that the spatial asymmetry of position capture occurs without attention but the spatial bias can be reduced by attention. Therefore, the underlying mechanism for the asymmetric spatial bias may be different from attentive tracking (Cavanagh, 1992 Science 257 1563-1565) and mislocalization during smooth pursuit (Brenner et al, 2001 Vision Research 41 2253-2259). PMID- 16042192 TI - The rod-and-frame effect: the whole is less than the sum of its parts. AB - Since the discovery of the influence of the tilted frame on the visual perception of the orientation perceived as vertical (VPV), the frame has been treated as a unitary object-a Gestalt. We evaluated the effect of 1-line, 2-line, 3-line, and 4-line (square frame) stimuli of two different sizes, and asked whether the influence of the square frame on VPV is any greater than the additive combination of separate influences produced by the individual lines constituting the frame. We found that, for each size, the square frame is considerably less influential than the additive combination of the influences of the individual lines. The results conform to a mass action rule, in which the lengths and orientations of the individual line components are what matters and the organization of the lines into a square does not-no higher-level Gestalt property is involved in the induction effect on VPV. PMID- 16042193 TI - Perceiving possibilities for action: on the necessity of calibration and perceptual learning for the visual guidance of action. AB - Tasks such as steering, braking, and intercepting moving objects constitute a class of behaviors, known as visually guided actions, which are typically carried out under continuous control on the basis of visual information. Several decades of research on visually guided action have resulted in an inventory of control laws that describe for each task how information about the sufficiency of one's current state is used to make ongoing adjustments. Although a considerable amount of important research has been generated within this framework, several aspects of these tasks that are essential for successful performance cannot be captured. The purpose of this paper is to provide an overview of the existing framework, discuss its limitations, and introduce a new framework that emphasizes the necessity of calibration and perceptual learning. Within the proposed framework, successful human performance on these tasks is a matter of learning to detect and calibrate optical information about the boundaries that separate possible from impossible actions. This resolves a long-lasting incompatibility between theories of visually guided action and the concept of an affordance. The implications of adopting this framework for the design of experiments and models of visually guided action are discussed. PMID- 16042194 TI - Effects of hand orientation and delay on the verbal judgment of haptically perceived orientation. AB - We examined the haptic perception of orientations of a single bar throughout the horizontal plane using a verbal response: participants were to assign a number of minutes to the orientation of a bar defined with respect to the stimulus table. Performance was found to be systematically biased. Deviations were consistent with, yet much smaller than, those resulting from haptic motor matching tasks. The size and direction of the deviations were found to correlate with hand orientation, and not to depend on spatial location per se, suggesting a role for hand-centred reference frames in biasing performance. Delaying the response by 10 s led to a small improvement only of right-hand perceptions, indicating different hemispheric involvement in processes involved in retaining and/or recoding of haptic orientation information. Also the haptic oblique effect was found with the current verbal response. Importantly, it was affected neither by hand orientation nor by delay, suggesting that the oblique effect is independent of the aforementioned deviations in orientation perception. PMID- 16042195 TI - Ambiguous depth planes: perceiving depth from motion. PMID- 16042196 TI - [Enhancement of the protective effect of SjC23 DNA vaccine against Schistosoma japonicum infection by immunostimulatory sequence]. AB - OBJECTIVE: To investigate the effect of immunostimulatory sequence on SjC23 DNA vaccine against Schistosoma japonicum infection. METHODS: SjC23 gene fragment was inserted into pcDNA3. 1-CpG to construct pcDNA3.1-SjC23/CpG. BALB/c mice in 4 groups were immunized intramuscularly 3 times at 2 week intervals, with 100 microg plasmid DNA per injection. Four weeks after the 3rd immunization, all mice were challenged with 45 +/- 1 cercariae of S. japonicum by abdominal skin penetration. After 45 days post-challenge, mice were perfused and the number of recovered worms and of eggs in liver was counted. Blood samples were collected from the tail vein of all mice 2 days before the 1st immunization and before challenge respectively. IgG, IgG1 and IgG2a in sera were detected. Three weeks after the 3rd inoculation, the spleen cells of 2 mice from each group were cultured and stimulated with ConA and recombinant peptide. The supernatant was collected to detect IL-2, IL-4 and IFN-gamma. Simultaneously, the cytotoxic activity was detected with 51Cr release assay in vitro. RESULTS: The worm reduction rate in SjC23 group and SjC23/CpG group was 28.1% and 35.1%, the hepatic egg reduction rate was 21.6% and 26.5%, respectively, compared with the control group. The level of protection in SjC23/CpG group was higher than that in SjC23 group (P<0.05). ELISA results indicated that mice immunized with pcDNA3.1 SjC23 and SjC23/CpG produced specific IgG to rSjC23, while mice immunized with pcDNA3.1 and pcDNA3.1-CpG did not. Mice in SjC23 group and SjC23/CpG group also produced IgG1 and IgG2a antibody isotypes, with the ratio of IgG2a/IgG1 10.1 and 12.2, respectively. In comparison with the control, the level of IL-2 and IFN gamma in mice immunized with pcDNA3.1-SjC23 and pcDNA3.1-SjC23/CpG was augmented. The cytotoxic activity of spleen cells from mice in SjC23/CpG group was augmented from 9.7% to 40.0% compared with that in SjC23 group. CONCLUSION: The study indicates that immunostimulatory sequence appears to increase the level of protection induced by immunization with pcDNA3.1-SjC23 vaccine. PMID- 16042197 TI - [Studies on the association of human leukocyte antigen class II alleles with advanced hepatosplenic schistosomiasis japonica]. AB - OBJECTIVE: To study the association of human leukocyte antigen class II (HLA-II) alleles with genetic susceptibility and resistance to advanced hepatosplenic schistosomiasis japonica. METHODS: The allelic types of HLA-DRB1, DPA1, DQA1 and DQB1 were detected by polymerase chain reaction with sequence-specific primers (PCR-SSP) technique in 46 patients with advanced hepatosplenic schistosomiasis, characterized with extensive liver fibrosis. Another 43 subjects with chronic schistosomiasis were used as control. The statistical significance of differences in allelic frequencies was determined by chi2 test. RESULTS: The frequencies of HLA-DRB1 x 04, DPA1 x 0103, DQA1 x 0601, DQB1 x 0201 in advanced patients were markedly higher than those in control group, while the frequencies of HLA-DQA1 x 0501 and DQB1 x 0601 in control group were higher than those in advanced patients. CONCLUSION: The study indicated that HLA-DRB1 x 04, DPA1 x 0103. DQA1 x 0601 and DQB1 x 0201 showing a positive, statistically significant (P<0.05) association with advanced hepatosplenic schistosomiasis japonica may be the susceptible genes, whereas HLA-DQA1 x 0501 and DQBH1 x 0601 may be more relevant to a resistance to the disease. PMID- 16042198 TI - [Case report: heavy infection of Strongyloides stercoralis complicated with meningioma and cholelithiasis]. PMID- 16042199 TI - [Clinical analysis on hepatic hydatid disease in Yili river valley]. AB - OBJECTIVE: To analyze the clinical diagnosis and treatment of hepatic hydatid disease and its epidemiological characteristics in Yili river valley. METHODS: Retrospective investigation was carried out on 2049 cases collected in 1993-2003. Clinical diagnosis was made by ways of intradermal test, serological test, ultrasound, X-ray, CT and/or MRI, majority of them received surgical operation. RESULTS: Among the 2049 cases, cystic hydatidosis occupied 96% (1965/2049), while 4% (84/2049) were alveolar hydatidosis. 99% (2034/2049) accepted surgery including hepatolobectomy, endocystomy and hydatidostomy in 302 cases (14.7%) without relapses. 754 cases (36.7%) received chemotherapy (praziquantel, albendazole) after surgical operation. The disease distributed in agri-pastoral areas along the valley. Local residents from different minorities had a close contact with dogs, 54% of the cases were females and 48% of the cases were in the group of 25-49 years old. The incidence tends to decline in the years. CONCLUSION: Hydatidosis is still an important health problem in the region. Further practices for improving treatment especially surgical intervention and for epidemiological investigation are needed. PMID- 16042200 TI - [Construction of monovalent and compound nucleic acid vaccines against Toxoplasma gondii with gene encoding p30]. AB - OBJECTIVE: To construct a monovalent gene vaccine pcDNA3.1-p30 and a compound gene vaccine pcDNA3.1-p30-ROP2 and assess the protective effect of the two vaccines against Toxoplasma gondii. METHODS: The sequences encoding p30 and ROP2 were amplified from the genomic DNA of T. gondii RH strain by polymerase chain reaction (PCR) and inserted into eukaryotic vector pcDNA3.1 to construct pcDNA3.1 p30 and pcDNA3.1-p30-ROP2. Mice were injected with the recombinant plasmid to observe the immunoprotectivity of the nucleic acid vaccine by using ELISA for detection of total IgG and observing the survival time after tachyzoites challenge. RESULTS: The recombinant plasmids pcDNA3.1-p30 and pcDNA3.1-p30-ROP2 were constructed. Mice in pcDNA3.1-p30-ROP2 group showed higher IgG (P < 0.05) and survived longer than those in pcDNA3.1-p30 group (P < 0.01) after challenged with T. gondii. CONCLUSION: Compound vaccine of genes from different stages of T. gondii elicits stronger immunoprotectivity in mice than a single gene vaccine. PMID- 16042201 TI - [Cloning and prokaryotic expression of transcriptional co-activator gene of Clonorchis sinensis and functional analysis of the expressed protein]. AB - OBJECTIVE: To construct prokaryotic recombinant plasmids of transcriptional co activator (TC) gene of Clonorchis sinensis, express and purify the recombinant protein and analyze its biological function. METHODS: A pair of primers was designed according to the known sequence of TC gene. The TC gene fragment was amplified by PCR. After purification and digestion with BamH I and Sal I, the TC gene was connected to the prokaryotic expression vectors, pGEX-4T-1 and pET30a(+). By cloning target gene into these vectors, pGEX-4T-1 and pET30a(+), prokaryotic recombinant plasmids of TC gene were constructed and transferred into E. coli BL21. The positive expressed recombinants were detected by SDS-PAGE and Western blotting. Immobilized metal (Ni2+) chelation affinity chromatography was used to purify His-TC produced by the expression of the recombinant protein pET30a(+)-TC. RESULTS: The recombinant plasmids, pGEX-4T-1-TC and pET30a(+)-TC, were constructed successfully. SDS-PAGE testified that the molecular weight of the recombinant protein was correct. Western blot analysis of GST-TC recombinant protein testified that the recombinant protein could be recognized by immunized rabbit serum, which means the protein is GST-immune active and the clone can express recombinant Clonorchis sinensis antigen. After affinity chromatography of the pET-TC protein, there was only one protein band with expected size on the SDS PAGE gel. CONCLUSION: The TC gene was screened from cDNA library of adult Clonorchis sinensis, cloned, expressed and purified. The purified protein of TC gene will be of importance for further research on the biological function of the gene. PMID- 16042202 TI - [Cloning and sequence analysis of Trichomonas vaginalis ferredoxin gene]. AB - OBJECTIVE: To construct a recombinant plasmid containing ferredoxin gene of Trichomonas vaginalis. METHODS: Total DNA was extracted from Trichomonas vaginalis with Chelex-100 method and used as templates for PCR. Primers were designed based on the published sequence of the ferredoxin gene and used to amplify the Trichomonas vaginalis gene using PCR method. The ferredoxin gene obtained by PCR technique was directionally cloned into plasmid pMD-18T simple vector. The constructed recombinant plasmid was transferred into E. coli JM109. The transformants were screened and identified by PCR and restriction analysis. The DNA sequence of the gene was determined by Sanger's method. RESULTS: The size of amplified ferredoxin gene was 306bp. The correct recombinant plasmid was isolated and confirmed by PCR and restriction analysis. The DNA sequence of cloned gene was the same as the published sequence. CONCLUSION: The ferredoxin gene was successfully amplified and cloned into plasmid pMD-18T simple vector. The cloned ferredoxin gene could be used to produce recombinant protein and for study of its function. PMID- 16042203 TI - [Fluctuation in the resistance of Plasmodium falciparum to chloroquine in China]. AB - OBJECTIVE: To investigate whether chloroquine-resistance of Plasmodium falciparum had changed after stopping or reducing the use of chloroquine as an antimalarial in Hainan and Yunnan provinces. METHODS: WHO standard in vitro microtest and 4 week in vivo test were used, assays were carried out in different time after stopping or reducing the use of chloroquine. RESULTS: In vitro test in Hainan indicated that the rate of chloroquine-resistant P. falciparum was 97.9% in 1981, and dropped to 26.7% in 1997 (P<0.01). The mean concentration of chloroquine for complete inhibition of schizont formation was 10.46 +/- 7.14 pmol/microl blood in 1981, decreased to 1.63 +/- 1.47 pmol/microl blood in 1997 (P<0.01). The proportion of samples taken from malaria cases that required high concentration (>6.4 pmol/microl blood) of chloroquine for complete inhibition of schizont formation was 83.3% in 1981 and only 6.7% in 1997 (P<0.01). In the 4-week in vivo test, the rate of chloroquine-resistant P. falciparum decreased from 84.2% in 1981 to 18.4% in 1997 (P<0.01). RIII cases accounted for 53.1% of the total resistant cases in 1981, and for 14.3% in 1997 (P<0.01). In vitro test in Yunnan revealed that the rate of chloroquine-resistant P. falciparum, the mean concentration of chloroquine for complete inhibition of schizont formation and the proportion of samples taken from malaria cases that required >6.4 pmol/microl blood of chloroquine for complete inhibition of schizont formation were 97.4%, 17.2 +/- 12.6 pmol/microl blood and 58.9% in 1981 respectively, and dropped to 70.4% (P< 0.01), 4.0 +/- 3.3 pmol/microl blood (P<0.01) and 16.6% (P<0.01) in 2002 respectively. CONCLUSION: The resistance of P. falciparum to chloroquine declined progressively after its use had been stopped or reduced in Hainan and Yunnan provinces. PMID- 16042204 TI - [Expression and antigenicity analysis of p46000 antigen from newborn larvae of Trichinella spiralis]. AB - OBJECTIVE: To express and analyze p46000 antigen from newborn larvae of Trichinella spiralis. METHODS: p46000 antigen gene was subcloned to the pET28a expression system by PCR. The recombinant transformant was induced by IPTG and the antigenicity was analyzed with ELISA and Western blotting. RESULTS: The molecular weight of the expressed protein was about Mr 48000. ELISA and Western blotting showed that this recombinant protein could be recognized by T. spiralis infected swine serum and rabbit anti-recombinant protein serum, and the rabbit anti-recombinant protein serum could recognize a Mr 46000 protein from newborn larvae of T. spiralis. CONCLUSION: p46000 recombinant antigen from newborn larvae of T. spiralis was expressed which shows a specific antigenicity. PMID- 16042205 TI - [Antigen analysis of Angiostrongylus cantonensis in different developmental stages]. AB - OBJECTIVE: To analyze the difference among antigens of Angiostrongylus cantonensis in different developmental stages and identify dominant diagnostic antigen for angiostrongyliasis. METHODS: Antigens of A. cantonensis in different developmental stages were analyzed by SDS-PAGE and immunoblot. RESULTS: The protein bands of all developmental stages were similar on SDS-PAGE. The Mr 40000, 50000, 66000 and 80000 antigens reacted not only with the sera of rats infected by A. cantonensis but also with the sera of normal rats. The Mr 104000 antigen could be discerned by sera of rats infected with A. cantonensis for 2 weeks. The Mr 32000 antigen could be recognized by sera of rats 2 weeks after infection, and the reaction became stronger with the infection continued. CONCLUSION: The Mr 40000, 50000, 66000 and 80000 antigens might result in the unspecific reaction in the immunodiagnosis of angiostrongyliasis using the crude antigen of A. cantonensis. The Mr 104000 of larva, Mr 33000 of adult females and Mr 32000 of the worms might be used as candidate antigens in early diagnosis and epidemiological survey of angiostrongyliasis. PMID- 16042206 TI - [Study on the biological characteristics of permethrin-resistant and susceptible strains of Culex pipiens pallens]. AB - OBJECTIVE: To compare the biological characteristics of Culex pipiens pallens between permethrin-resistant strain and the susceptible strain. METHODS: In laboratory, the biological characteristics about bloodsucking, reproduction and development of permethrin-resistant and susceptible strains were observed and recorded. Life table of the experimental populations was constituted. RESULTS: The bloodsucking rate of resistant strain was lower than susceptible strain and the difference was significant. The life-span of adults of resistant strain and the developmental duration of eggs, larvae and pupae were longer than susceptible strain and the difference was significant. The death rate of eggs of resistant strain was higher than that of susceptible strain and the difference was significant. The permethrin-resistant strain showed a fitness value of 0.58 relative to the susceptible strain. CONCLUSION: The permethrin resistance of Culex pipiens pallens may result in a disadvantage to its reproduction and development. PMID- 16042207 TI - [Construction of prokaryotic expression vector of the fusion gene IFN-alpha1b/CSP II and expression in E. coli]. AB - OBJECTIVE: To Construct the prokaryotic expression vector of the fusion gene IFN alpha1b/CSP II. METHODS: IFN-alpha1b was amplified from the human genomic DNA by PCR and cloned into prokaryotic expression vector pGEX-4T-1. The recombinant plasmid pGEX-4T-1/IFN-alpha1b was constructed. Circumsporozoite protein II (CSP II) was amplified from the Plasmodium falciparum genomic DNA by PCR and was cloned into the prokaryotic expression vector pGEX-4T-1. The recombinant plasmid pGEX-4T-1/CSPII was constructed. IFN-alpha1b was cut from the recombinant plasmid pGEX-4T-1/IFN-alpha1b digested with BamH I and EcoR I and ligated with the recombinant plasmid pGEX-4T-1/CSP II also digested with BamH I and EcoR I. The recombinant prokaryotic plasmid pGEX-4T-1/IFN-alpha1b/CSP II was constructed. The fusion gene IFN-alpha1b/ CSP II was expressed in E. coli by IPTG. RESULTS: The prokaryotic expression vector pGEX-4T-1/IFN-alpha1b, pGEX-4T-1/CSP II and pGEX-4T 1/IFN-alpha1b/CSP II were identified by PCR, enzyme digestion and gene sequencing. The expressed fusion protein/IFN-alpha1b/CSP I in E. coli was identified by SDS-PAGE and Western blot. CONCLUSION: The prokaryotic expression vector of the fusion gene IFN-alpha1b/CSP II was successfully constructed, which was then expressed in E. coli. PMID- 16042208 TI - [Detection of Pneumocystis carinii DNA in rats by PCR-ELISA]. AB - OBJECTIVE: To establish a PCR-ELISA and evaluate its use in detecting DNA of Pneumocystis carinii (P. c) in rat model. METHODS: SD rats and Wistar rats were used in the experiment. P. c DNA from rat lung tissue and BALF was amplified by PCR. The amplified products were visualized by ethidium bromide (EB) staining after agarose gel electrophoresis or detected by ELISA. The results were compared with that by Giemsa stain. RESULTS: The positive rate in the two species of rats by the two methods was 96.4% and 100% in lung tissue, 96.4% and 100% in BALF, respectively, with no significant difference (P>0.05). Giemsa positive samples were all positive by PCR-ELISA. The negative control group had one positive by ELISA in lung tissue and BALF respectively. CONCLUSION: PCR-ELISA shows a high sensitivity and specificity in detecting the DNA of Pneumocystis carinii, which is a secure and easy use method. PMID- 16042209 TI - [Genetic characteristics of Trichomonas vaginalis]. PMID- 16042210 TI - [Research progress on the adhesion course of Trichomonas vaginalis]. PMID- 16042211 TI - [An introduction to the Internet resources on acarology]. PMID- 16042212 TI - [Cell apoptosis and necrosis in the development of Trichinella spiralis larvae]. PMID- 16042213 TI - [Imaging observation on 47 cases of paragonimiasis westermani]. PMID- 16042214 TI - [Control of anaphylactic shock during puncture of hydatid cyst in liver]. PMID- 16042215 TI - [Effect of feces management and chemotherapy in the control of cattle schistosomiasis in lake region]. PMID- 16042216 TI - Acute partial transverse myelitis with normal cerebral magnetic resonance imaging: transition rate to clinically definite multiple sclerosis. AB - OBJECTIVE: To determine the long-term risk of developing clinically definite multiple sclerosis (CDMS) in patients with acute partial transverse myelitis (APTM) and normal cerebral magnetic resonance imaging (MRI) scans. METHODS: We retrospectively studied 30 consecutive patients with clinical evidence of APTM. Patients with symmetric severe acute transverse myelitis were considered to have complete transverse myelitis and were excluded. All patients underwent spinal and cerebral MRIs, 13 underwent cerebrospinal fluid analysis and 11 patients underwent evoked potential studies. Various other studies were performed to assess for connective tissue disease and causes of APTM other than demyelinating disease. RESULTS: After an average follow-up of 61 months, all laboratory and clinical evidence, including relapse history, indicated that three patients developed lesions on cerebral MRI and could be classified as CDMS by either Poser criteria (two patients) or MacDonald criteria (one patient). Relapses limited to the spinal cord seen clinically were seen in 14/30 (46.6%) patients. Oligoclonal bands were seen in 8/13 (62%) patients; one patient transitioned to CDMS. Unifocal lesions of the cord were seen in 19/30 (63%) patients, multifocal lesions were seen in 8/30 (27%) and 3/30 (10%) had negative MRIs. The three patients who converted to CDMS did so within five years of the onset of myelitis. CONCLUSION: APTM with normal cerebral MRI had a low rate of conversion to CDMS in this long-term study. To date, there have been only a few follow-up studies that have addressed this issue. PMID- 16042217 TI - A comparative audit of anticardiolipin antibodies in oligoclonal band negative and positive multiple sclerosis. AB - It has been suggested that multiple sclerosis (MS) patients with positive anticardiolipin antibodies (ACLA) have some atypical features, including absent oligoclonal bands (OCB) in the cerebrospinal fluid (CSF). Our aim was to compare the frequencies of ACLA and related laboratory and clinical features in OCB negative (OCB-) and positive (OCB+) MS patients. We compared 41 OCB- patients attending a MS Clinic in a tertiary referral center, with 206 OCB+ patients. ACLA, anti-beta2-glycoprotein and other autoantibodies, lupus anticoagulant and coagulation markers were measured. We found a higher frequency of ACLA in OCB- patients, 18/41 versus 33/206 in OCB+ patients (P<0.0001). OCB- patients had more progressive MS than OCB+ subjects. There were no differences in age, sex, Expanded Disability Status Scale (EDSS) score, antiphospholipid syndrome symptoms between the groups. ACLA+ MS patients were more frequently in the OCB- group. Although this may suggest that they represent a special subgroup of MS, no other clinical or laboratory findings distinguish the groups. Although OCB- MS patients may be thought to be less active immunologically, this study shows they have more frequently ACLA than OCB+ patients. OCB- MS patients in our cohort do not appear to have a more benign form of MS, as has previously been suggested. PMID- 16042218 TI - Autoantigen-pulsed dendritic cells constitute a beneficial cytokine and growth factor network in ameliorating experimental allergic encephalomyelitis. AB - Injection of myelin basic protein (MBP)-pulsed dendritic cells (DC) into healthy rats, as we reported before and observed in this study, did not induce clinical experimental allergic encephalomyelitis (EAE), but effectively protected the rats from subsequent EAE induction. The mechanisms by which MBP-pulsed DC mediate immune protection are not completely understood. In the present study, we mainly explored the dynamic change of cytokine and growth factor mRNA expression in spinal cords after subcutaneous injection of MBP-pulsed and unpulsed DC. The expression of interleukin (IL)-1, interferon-gamma and tumour necrosis factor alpha as well as programmed death ligand (PDL)-1, PDL-2, signal transducer and activator of transcription (STAT)4, STAT6, matrix metalloproteinase (MMP)-9 and tissue inhibitor of metalloproteinases (TIMP)-2 was increased on day 0 postimmunization (p.i.). The increase of IL-12 expression was observed on day 7 p.i., while the increase of IL-10 expression mainly occurred on day 14 p.i. Except downregulation of insulin-like growth factor-1, the expression of brain derived neurotrophic factor, ciliary neurotrophic factor, fibroblast growth factor (FGF)-2 and platelet-derived growth factor (PDGF)-B/C as well as nerve growth factor receptor (NGF-R), FGF receptor, PDGF-R-alpha and beta was elevated on day 0 p.i., while the increase of TIMP and NGF was observed on days 0 and 7 p.i. There were no significant differences on MMP-2, spinal cord-derived growth factor and PDGF-A mRNA expression. In line with the suppression of EAE induced by MBP-pulsed DC, the dynamic change of cytokines and growth factors in spinal cords should constitute a beneficial microenvironment against EAE. PMID- 16042219 TI - Monthly brain magnetic resonance imaging scans in patients with clinically isolated syndrome. AB - We investigated if monthly gadolinium (Gd)-enhanced magnetic resonance imaging (MRI) can assist the clinician in anticipating the diagnosis of multiple sclerosis (MS) in the very first few months following a clinically isolated syndrome (CIS). A consecutive series of CIS patients with > or = 3 T2-weighted (T2W) hyperintense brain MRI lesions suggestive of MS were followed up for the first six consecutive months after enrollment with monthly triple-dose Gd enhanced brain MRI scan. MRI conversion to MS was defined by the presence of either > or = 1 new Gd-enhancing lesion or > or = 1 new T2W lesions in the subsequent MRI scan. Sixty patients were included. Of them, 30 (50%) had at least one Gd-enhancing lesion on the baseline MRI scan. After three months, MRI conversion to MS was observed in 80% and 62% of patients based on the appearance of > or = 1 new T2 lesion and > or = 1 new Gd-enhancing lesions, respectively. The presence of > or = 1 new T2W lesion was observed in 90% and 82% of patients who had, at baseline, a Gd-positive MRI scan and dissemination in space based on the new McDonald's criteria, respectively The rate of MRI conversion remained almost stable in the last two MRI scans. Our study suggests that the majority of CIS patients with an abnormal baseline scan showed an MRI conversion to MS after three months. The model of six months as the optimal interval for repeating MRI exam is not supported by the present data. PMID- 16042220 TI - Sequential magnetic resonance imaging follow-up of multiple sclerosis before the clinical phase. AB - There is much evidence of the importance of the preclinical phase of multiple sclerosis (MS). However, apart from a recent report of the incidental discovery of a case of primary progressive MS, there are no data on the sequential magnetic resonance imaging (MRI) follow-up of patients before the clinical phase. We report the incidental discovery of white matter changes on MRI and their follow up in a patient three years before the first neurological event (optic neuritis). A 34-year-old woman presented with headaches and depression after her young daughter had been involved in a car accident and spent two weeks in intensive care. The woman's general practitioner performed a brain MRI, which revealed multiple T2-weighted hypersignals suggesting MS. During the next three years, clinical examination remained normal but we observed new T2 lesions and/or new enhanced T1 lesions after gadolinium infusion on the four successive MRIs. Thirty seven months after the first MRI, the patient developed a right optic neuritis. The diagnosis of MS was made according to space and time dissemination on MRI criteria. We proposed a treatment with Interferon Beta 1a (Avonex). PMID- 16042221 TI - Reappraisal of Lhermitte's sign in multiple sclerosis. AB - Lhermitte's sign (LS) is strongly linked to multiple sclerosis (MS). Our aim is to reassess its frequency, natural history, various characteristics and neuroradiological correlation in a cohort of MS patients attending our specialized MS clinic and to propose a working definition. Consecutive patients with CDMS and normal controls were interviewed using a structured questionnaire. Cervical MRIs were reviewed when available. There were 300 MS patients and 100 normal controls. Forty-one per cent of the patients and none of the controls reported having LS during the course of their illness. In 53% of those who reported LS, it started in the first three years of the illness and began as an isolated symptom in 64% and was polysymptomatic in 36%. In all patients LS was a short-lasting sensation in all patient who experienced it and was mostly stereotyped in individual patients. Characteristics varied widely between patients. Forty-three patients had cervical MRIs; 17 out of 18 patients who reported LS had abnormalities, whereas only 13 out of the 25 with no LS had abnormalities. The results indicate that LS is highly prevalent in MS, is commonly stereotyped in individual patients, has a variable natural course and correlates significantly with cervical MRI abnormalities. A working definition is proposed. PMID- 16042222 TI - Multiple sclerosis in Tayside, Scotland: detection of clusters using a spatial scan statistic. AB - Debate continues over the relative importance of genetic factors over infectious agents in the aetiology of multiple sclerosis (MS). Detection of clusters of MS in space and time in the Tayside region of Scotland, UK would provide valuable evidence for the movement of infectious agents into a genetically susceptible population. A spatial scan statistic was used to detect, locate and provide a robust statistical test of any clusters found, without prior knowledge of their location or size. This was applied to a population-based MS register for the Tayside region of Scotland from 1970 to 1997, allowing for age at symptom onset, gender, population density and social deprivation. There were a total of 772 cases during the study period; an annual incidence of 72 per 100000. The mean age of symptom onset was 35.7 (SD = 10.5) and 73.8% of cases were women. There was a general increase in cases over time probably reflecting gradually better detection and diagnosis. There was a peak around the mid-1990s and some evidence of periodicity. There was a highly significant temporal cluster between 1982 and 1995 (P = 0.002) for the whole region. Additionally, a significant spatial cluster for the time period 1993-1995 was found centred in the rural area south west of Perth (P=0.016). Significant temporal and spatial-temporal clusters are consistent with exogenous factors contributing to the distribution of MS in Tayside, Scotland. PMID- 16042223 TI - Eight-year immunogenicity and safety of interferon beta-1a-Avonex treatment in patients with multiple sclerosis. AB - An open-label extension study of the phase III trial of intramuscular interferon beta-1a (IFNbeta-1a-Avonex) was conducted to evaluate the immunogenicity and safety of IFNbeta-1a-Avonex over six years in patients with relapsing multiple sclerosis (MS). Patients who participated in the pivotal phase III study were offered enrolment; entry was also open to patients who had not participated. All patients received IFNbeta-1a-Avonex 30 microg intramuscularly once weekly for six years, for a treatment duration of up to eight years in patients who received IFNbeta-1a-Avonex in the phase III trial. Serum levels of IFNbeta antibodies were measured every six months using a screening enzyme-linked immunosorbent assay (ELISA) followed by an antiviral cytopathic effect assay to detect neutralizing antibodies (NAbs) in serum samples positive on ELISA. The incidence of adverse events and laboratory test results assessed safety. Of 382 total patients, 218 had participated in the phase III study (103 placebo, 115 IFNbeta-1a-Avonex) and 164 had not participated; 24 of the 164 were IFNbeta-naive. At baseline, 281 patients were negative for IFNbeta antibodies (NAb-). NAbs (titre > or = 20) developed at any time over six years in 5% of these patients. Of 140 patients who had been on IFNbeta-1b-Betaseron, 49 were positive for NAbs (NAb+) at baseline; 11 of 115 who had been on IFNbeta-1a-Avonex were NAb+ at baseline. Thirty-nine of 49 patients who had been on Betaseron and were NAb+ had titres < 100; 36 of these 39 seroconverted to NAb- while on IFNbeta-1a-Avonex, with a median time of approximately six months. Ten patients who had been on Betaseron had NAb titres > or = 100; five remained NAb+ during six years on IFNbeta-1a-Avonex and five seroconverted to NAb-, but only after at least two years. Five patients who had been on IFNbeta-1a-Avonex during the clinical trial were NAb+ with titres < 100 at baseline; four seroconverted to NAb-, with a median time of two to three years. Six patients who had been on IFNbeta-1a-Avonex had NAb titres > or = 100; five of these remained NAb+ at six years. No patient with a NAb titre > 1000 seroconverted to NAb-, whether initially treated with IFNbeta-1a-Avonex or Betaseron. Adverse events were similar to those observed in the pivotal phase III trial. Results from this trial indicated that IFNbeta-1a-Avonex was associated with a low incidence of NAbs and was well tolerated for up to eight years. Further, the results indicate that persistence of NAbs is dependent on titre and IFNbeta product. PMID- 16042224 TI - Disease-modifying drugs in childhood-juvenile multiple sclerosis: results of an Italian co-operative study. AB - OBJECTIVE: Immunomodulatory drugs (IDs) (interferon beta (IFNgamma) and glatiramer acetate (GA)) reduce relapse rate and disease progression in relapsing remitting multiple sclerosis (RRMS) but extensive data are not available on the effectiveness and tolerability of these drugs in childhood or adolescence. The aim of this study was to evaluate the impact of IFNbeta and GA in MS patients treated before 16 years of age. METHODS: A research group (Immunomodulatory Treatment of Early onset MS (ITEMS)) was promoted in Italy to collect a large series of patients affected by clinically definite and RRMS and treated with IDs before 16 years of age. Fifteen centres recognized subjects suitable for inclusion: 76 patients (52 females) were collected with a mean age at onset of 12.4 (SD 2.5) years, a mean disease duration of 18.6 (SD 14.7) and a relapse rate of 3.1 (SD 2.9). RESULTS: Results were evaluated in 65 (45 females) subjects with a pretreatment and a treatment duration >3 months: 38 were treated with IFNbeta 1a once weekly (Avonex), 18 with IFNbeta three times weekly (16 with Rebif, 2 with Betaferon) and nine with GA (Copaxone). The mean pretreatment period was respectively 20, 18 and 9.2 months. The treatment duration lasted respectively 23.3, 40.7 and 33.3 months. The mean annualized relapse rate decreased dramatically during the treatment: from 2.4 to 0.4 in the Avonex group, from 3.2 to 0.8 in the Rebif-Betaferon group and from 2.8 to 0.25 in the GA group. The mean final EDSS scores were respectively (in brackets the initial scores): 1.3 (1.4), 1.6 (1.8) and 0.6 (1.1). In the whole group, the final score was unchanged or reduced in all subjects except eight. Clinical side effects were recorded in 41/65 subjects (mainly in subjects treated with IFNbeta), abnormal laboratory findings were observed in 13/65 subjects: they were transient in most cases. IFNgamma was stopped in six cases: in four because of inefficacy and in two cases because of side effects. CONCLUSIONS: Sixty-five clinically definite MS subjects were treated during childhood or adolescence with IDs. The treatment reduced the relapse rate and the progression of the disease in most cases. Side effects were common in subjects treated with IFNbeta but were well tolerated in most cases. PMID- 16042225 TI - The mechanism of action of methylprednisolone in the treatment of multiple sclerosis. AB - Methylprednisolone plays an important role in the current treatment of multiple sclerosis (MS), particularly in the acute phase of relapse. It acts in various ways to decrease the inflammatory cycle including: dampening the inflammatory cytokine cascade, inhibiting the activation of T cells, decreasing the extravasation of immune cells into the central nervous system, facilitating the apoptosis of activated immune cells, and indirectly decreasing the cytotoxic effects of nitric oxide and tumor necrosis factor alpha. This paper reviews the most recent observations on these mechanisms both to understand the disease mechanism and its treatment. As more becomes known about these mechanisms, it may become possible to design treatment regimes that are more specific towards both the individual and the disease state. PMID- 16042226 TI - MRI results from the European Study on Intravenous Immunoglobulin in Secondary Progressive Multiple Sclerosis (ESIMS). AB - BACKGROUND: Monthly application of high-dose intravenous immunoglobulin (IVIG) to patients with secondary progressive multiple sclerosis (MS) showed no clinical benefit in the European Study on Immunoglobulin in MS (ESIMS). Magnetic resonance imaging (MRI) results may provide insights into the morphologic consequences of such treatment. METHODS: A total of 318 patients (mean age 44 +/- 7 years) were enrolled in 31 European and Canadian centres and treated monthly with 1 g/kg body weight of IVIG or equivalent amounts of albumin 0.1% for 27 months. MRI was performed at baseline and after 12 and 24 months and comprised of conventional dual-echo T2-weighted and T1-weighted scans before and after application of 0.1 mmol/kg Gd-DTPA. RESULTS: Similar to clinical variables, MRI measures at baseline were well comparable between treatment groups except for a somewhat lower mean number of contrast-enhancing lesions and number of active scans in IVIG-treated patients. Over the trial period there was almost no change of the T2-lesion load and the 'black hole' volume in both treatment groups and the cumulative number of contrast-enhancing lesions were similar. There was only a trend for fewer new or enlarged T2-lesions in IVIG patients, which disappeared after correction for the imbalance in the number of contrast-enhancing lesions at baseline. Brain volume in terms of a partial cerebral fraction decreased significantly less with IVIG than placebo treatment (final visit: -0.62 -/+ 0.88% versus -0.88 +/- 0.91%; P=0.009). This difference remained statistically significant with correction for active lesions at baseline (P=0.02) and was seen primarily in male patients and those with an Expanded Disability Status Scale score > or = 6 and no relapses in the two years before the study. CONCLUSION: The absence of significant differences in conventional MRI measures between both treatment groups parallels the negative clinical results of ESIMS. The causes for and possible long-term clinical effects of a lower rate of brain volume loss in IVIG patients should be explored further. PMID- 16042227 TI - Serum MMP-9/TIMP-1 and MMP-2/TIMP-2 ratios in multiple sclerosis: relationships with different magnetic resonance imaging measures of disease activity during IFN beta-1a treatment. AB - Matrix metalloproteinase-9 (MMP-9) is involved in blood-brain barrier (BBB) disruption in active multiple sclerosis (MS), while MMP-2 seems to be associated with the chronic progressive phase of the disease. Recombinant interferon beta-1a (rIFNbeta-1a) is effective in restoring the BBB. We studied the relationships between serum MMP-9, MMP-2, TIMP-1 and TIMP-2 and different magnetic resonance imaging (MRI) measures of disease activity in MS patients during treatment with rIFNbeta-1a. Twenty-one relapsing-remitting (RR) MS patients underwent longitudinally simultaneous blood withdrawals and MRI (before and after standard dose (SD) and triple dose (TD) of gadolinium (Gd)) examinations before and during 48 weeks of rIFNbeta-1a (Rebif 22 mcg three times a week) treatment. Serum MMP-9, MMP-2, TIMP-1 and TIMP-2 were measured, MMP-9 to TIMP-1 and MMP-2 to TIMP-2 ratios were calculated and the numbers of Gd-SD, Gd-TD, new-Gd-SD, new-Gd-TD and new-T2 lesions counted. Serum MMP-9/TIMP-1 ratio (P < 0.0001), as well as the numbers of 'active' lesions (P ranging from 0.0004 to 0.005) decreased during treatment Moreover, serum MMP-9/TIMP-1 ratio proved to be a good positive predictor (estimate = 0.85; P < 0.05) of the numbers of MRI Gd-TD active lesions. These data confirm that serum MMP-9/TMIP-1 ratio may be viewed as a reliable marker and may be predictive of MRI activity in RR MS. PMID- 16042228 TI - The distribution of the magnetic resonance imaging response to glatiramer acetate in multiple sclerosis. AB - We investigated the distribution of the magnetic resonance imaging (MRI)-measured response to glatiramer acetate (GA) treatment in multiple sclerosis (MS) using data from a clinical trial of relapsing-remitting (RR) MS. A fixed and a random effects model were used to quantify the between-patient heterogeneity in treatment response, expressed as new enhancing lesion percentage reduction. In 95% of the patients, lesion reduction due to treatment was estimated to range between -20% and -54%, indicating a rather homogeneous effect of GA on MRI measured disease activity in RRMS. PMID- 16042229 TI - Health-related quality of life in multiple sclerosis: psychometric analysis of inventories. AB - OBJECTIVE: Measuring health-related quality of life (HRQoL) enables attainment of a comprehensive picture of chronic patients' conditions. The aim was to gain insight into HRQoL as viewed by Israeli multiple sclerosis (MS) patients, with an eye to its cross-cultural applicability, using psychometric analysis and a regression-based procedure. METHODS: The reliability and validity of the MSQoL-54 was assessed in 215 Israeli MS patients. Subsequently a novel method in QoL research for assessing the relative importance to patients of different aspects of the construct, comparing these between males and females, was applied. Using factor analysis, the degree to which the MSQoL-54 dimensions are applicable in our population and the existence of a separate dimension of Fatigue were tested. Finally, structural equation modelling (SEM) was used to examine relationships between QoL dimensions. RESULTS: The MSQoL-54 showed reliabilities and validities comparable to those found in other populations. Females attach importance to emotional aspects of QoL, while males are more concerned with physical aspects. The distinction between Emotional and Physical dimensions emerged from SEM, as did a distinction between Fatigue and Energy, the former being primarily Physical, and the latter Emotional. SEM indicated a reciprocal causality between Physical and Emotional dimensions. CONCLUSIONS: Based on internal structure, reliability and validity, the MSQoL-54 is applicable to Israeli MS patients. However, the scale's Social Function dimension is indecisively defined, which requires further study. QoL research in MS should distinguish between emotional and physical Fatigue. Assessing importance of QoL dimensions should inform clinicians in tailoring treatment to the individual patient. PMID- 16042230 TI - Physical activity and multiple sclerosis: a meta-analysis. AB - Using meta-analytic procedures, this study involved a quantitative synthesis of the difference in physical activity among individuals with multiple sclerosis (MS) compared with nondiseased and diseased populations and then examined factors (i.e., moderators) that explain variation in the overall difference in physical activity. We searched MEDLINE, PsycINFO and Current Contents Plus using the key words physical activity, exercise and physical fitness in conjunction with multiple sclerosis; conducted a manual search of bibliographies of the retrieved papers; and contacted study authors about additional studies. Overall, 53 effects were retrieved from 13 studies with 2360 MS participants and yielded a weighted mean effect size (ES) of -0.60 (95% CI = -0.44, -0.77). The weighted mean ES was heterogenous, Q = 1164.11, df = 52, P < 0.0001. There were larger effects with objective versus self-report measures of physical activity, nondiseased versus diseased populations and primary progressive versus relapsing remitting MS. The cumulative evidence suggests that individuals with MS are less physically active than nondiseased, but not diseased, populations. PMID- 16042231 TI - Emotional responses of children and adolescents to parents with multiple sclerosis. AB - The effect of a chronic illness of one parent on children is determined by a complicated interaction of various emotional components. Our focus was on the children's and adolescent's emotional reactions and feelings towards their multiple sclerosis (MS)-affected parents, including: degree of responsibility, obligation and concern, yielding behaviour and active protection, fear and anxiety related to the state of illness, their sense of burden in connection with household tasks and errands, and anger. Fifty-six children, ages 10-18, each having a parent with MS, were examined. The results were compared to a control group of 156 age-matched children with healthy parents. Feelings were examined by means of a questionnaire previously constructed by us. We found that children of parents with MS felt more responsibility and obligation than children of healthy parents. They also exhibited more yielding behaviour, more fear and anxiety related to states of illness, a greater sense of burden and a greater degree of anger. We consider the interaction between the sex of the parent and the sex of the child in connection with these feelings and discuss the implications of the 'parental child' role of these children. PMID- 16042232 TI - The association between cognitive impairment and physical disability in multiple sclerosis. AB - BACKGROUND: The association between cognitive impairment and physical disability was examined in a larger, more representative sample of patients with multiple sclerosis (MS) than in previous studies. METHOD: Two hundred and fifty-three patients attending an MS clinic were assessed with respect to physical disability using the Expanded Disability Status Scale and cognitive impairment using a battery of neuropsychological tests. RESULTS: Physical disability correlated with duration of disease; cognitive impairment did not. Virtually all measures derived from the cognitive battery were significantly correlated with physical disability. Three measures of speeded information processing and one involving delayed recall of verbal material were unique predictors of disability status. The relationship between cognitive impairment and physical disability was equivalent for patients with shorter (< 3 years) versus longer (> 10 years) disease duration. Cognitive impairment correlated with the rate of disability progression as reflected by the progression index. CONCLUSION: Cognitive impairment is more closely associated with physical disability than most previous studies indicate. This relationship appears to be stable throughout the duration of MS, although this conclusion is qualified by the cross-sectional design of the study. Further attention should be paid to cognitive impairment as a possible predictor of the rate of patients' physical decline. PMID- 16042233 TI - A review of stress-relapse interactions in multiple sclerosis: important features and stress-mediating and -moderating variables. AB - Studies do not provide a consensus opinion of the relationship between stress and relapse in relapsing-remitting multiple sclerosis (RRMS). Few studies have defined the critical features of these stressful situations, or examined the role of stress-mediating and -moderating variables. Available evidence indicates that the relationship between life stress and relapse is complex, and is likely to depend on factors such as stressor chronicity, frequency, severity and type, and individual patient characteristics such as depression, health locus of control and coping strategy use. Little is known about how these factors, individually or in combination, are related to MS disease activity. Viral infections are also likely to precipitate relapse in MS, and significant life-stress may further enhance this relationship. The nature and strength of these interrelationships have strong clinical implications. MS patients are particularly vulnerable to a deteriorating cycle of stressful life events, illness episodes and disability. Timely multidisciplinary care interventions aimed at both minimizing psychological distress and physical symptoms may halt this downward reciprocal cycle. Little is known of the pathogenesis of these putative stress-induced changes in disease activity, and almost all stressor studies suffer from some biases or limitations. PMID- 16042234 TI - Motor determinants of gait in 100 ambulatory patients with multiple sclerosis. AB - A prospective analysis of gait and strength parameters was performed in 100 patients diagnosed with MS and pyramidal involvement admitted in a rehabilitation unit The patients were divided into two groups based on their ability to walk in daily life (nonassisted or cane-assisted gait) and into four clinical subgroups depending on associated involvements such as sensory loss or cerebellar ataxia. Twenty healthy subjects were studied as a control group. Gait parameters were evaluated with a Locometre and muscle strength with an isokinetic dynamometer. The results showed that the average velocity and strength of the hamstring and quadriceps were strongly correlated and reduced in the MS group in comparison with the control, and in the cane-assisted group compared with the nonassisted group. Gait velocity tended to be more correlated to hamstring strength in the nonassisted group with a determination coefficient (r2) reaching a value of 0.44 in the sensory subgroup. These findings provide evidence that a correlation between strength reduction and gait impairment is obvious whatever the clinical form in patients with MS. This correlation is higher with hamstrings but may change depending on the disability level and the clinical form. This could be taken into account in the individual assessment of further rehabilitation programmes. PMID- 16042235 TI - Anti-myelin antibodies do not allow earlier diagnosis of multiple sclerosis. AB - This study investigates whether the presence of serum and plasma anti-myelin oligodendrocyte glycoprotein (MOG) and anti-myelin basic protein (MBP) in patients presenting with a clinically isolated syndrome compatible with demyelination (CIS) predicts early conversion to multiple sclerosis (MS). Forty seven patients with CIS (46 with optic neuritis) had anti-MOG and anti-MBP antibodies analysed at baseline, and clinical and magnetic resonance imaging assessments. There was no evidence that the MS status based on either the McDonald or Poser criteria relates to the antibody status. PMID- 16042236 TI - No evidence for MSRV viraemia and glial cell death in acute optic neuritis. PMID- 16042237 TI - Danish guidelines for small-scale constructed wetland systems for onsite treatment of domestic sewage. AB - The Danish Ministry of Environment and Energy has passed new legislation that requires the wastewater from single houses and dwellings in rural areas to be treated adequately before discharge into the aquatic environment. Therefore official guidelines for a number of onsite treatment solutions have been produced. These include guidelines for soakaways, biological sand filters, technical systems as well as different types of constructed wetland systems. This paper summarises briefly the guidelines for horizontal flow constructed wetlands, vertical flow constructed wetlands, and willow systems with no outflow and with soil infiltration. There is still a lack of a compact onsite solution that will fulfil the treatment classes demanding 90% removal of phosphorus. Therefore work is presently being carried out to identify simpler and robust P-removal solutions. PMID- 16042238 TI - How to treat raw sewage with constructed wetlands: an overview of the French systems. AB - The development of vertical flow constructed wetlands treating raw wastewater in France has proved to be very successful over the last 20 years. In view of this a survey was carried out on more than 80 plants in order to study their performance and correct the design if necessary. This study shows that such systems perform well in terms of respecting the goals of both low level outlet COD and SS and nitrification. Pollutant removal performance in relation to the loads handled and the specific characteristics of the plants were investigated. Nitrification is shown to be the most sensitive process in such systems and performance in relation to sizing is discussed. Such systems, if well designed, can achieve an outlet level of 60 mg L(-1) in COD, 15 mg L(-1) in SS and 8 mg L(-1) in TKN with an area of 2 - 2.5 m2.PE(-1). The sludge deposit on the first stage must be removed after about 10-15 years. PMID- 16042239 TI - Constructed wetlands for CSO treatment: an overview of practice and research in Germany. AB - Vertical flow treatment wetlands have been developed as very useful tools for treatment of combined sewage overflow. Several systems have been in operation for over 15 years. Based on recent research work, new technical guidelines now recommend systems with a drained filter of sand 0/2 mm and a throttled outflow. COD, NH4-N and SS removal rates of 85-99% can be expected from this type of filter. SS loadings that are too high and very long or frequent inundation affect the performance adversely. Information for successful long-term operation were derived from various existing plants. PMID- 16042240 TI - Constructed wetlands in UK urban surface drainage systems. AB - This paper presents the outcome of an inventory of planted wetland systems in the UK which are classified according to land use type and are all examples of sustainable drainage systems. The introduction of constructed wetlands to treat surface runoff essentially followed a 1997 Environment Agency for England and Wales report advocating the use of "soft engineered" facilities including wetlands in the context of sustainable development and Agenda 21. Subsequently published reports by the UK Construction Industry Research and Information Association (CIRIA) have promoted the potential benefits to both developer and the community of adopting constructed wetlands and other vegetated systems as a sustainable drainage approach. In addition, the UK Environment Agency and Highways Agency (HA) have recently published their own design criteria and requirements for vegetative control and treatment of road runoff. A case study of the design and performance of a constructed wetland system for the treatment of road runoff is discussed. The performance of these systems will be assessed in terms of their design criteria, runoff loadings as well as vegetation and structure maintenance procedures. The differing design approaches in guidance documents published in the UK by the Environment Agency, CIRIA and HA will also be evaluated. PMID- 16042241 TI - Free water surface wetlands for wastewater treatment in Sweden: nitrogen and phosphorus removal. AB - In South Sweden, free water surface wetlands have been built to treat wastewater from municipal wastewater treatment plants. Commonly, nitrogen removal has been the prime aim, though a significant removal of tot-P and BOD7 has been observed. In this study, performance data for 3-8 years from four large (20-28 ha) FWS wetlands have been evaluated. Two of them receive effluent from WWTP with only mechanical and chemical treatment. At the other two, the wastewater has also been treated biologically resulting in lower concentrations of BOD7 and NH4+-N. The wetlands performed satisfactorily and removed 0.7-1.5 ton N ha(-1) yr(-1) as an average for the time period investigated, with loads between 1.7 and 6.3 ton N ha(-1)yr(-1). Treatment capacity depended on the pre-treatment of the water, as reflected in the k20-values for N removal (first order area based model). In the wetlands with no biological pre-treatment, the k20-values were 0.61 and 1.1 m month(-1), whereas for the other two they were 1.7 and 2.5 m month(-1). P removal varied between 10 and 41 kg ha(-1) yr(-1), and was related to differences in loads, P speciation and to the internal cycling of P in the wetlands. PMID- 16042242 TI - Using a compact combined constructed wetland system to treat agricultural wastewater with high nitrogen. AB - The objectives of this study were to find appropriate conditions for nitrogen removal by a compact combined constructed wetland system and to evaluate the removal rate constant in a tropical climate. This study will present suitable operating conditions for a combined system to treat pig farm wastewater containing high ammonia-nitrogen. Four laboratory-scale combined constructed wetland units (0.5 x 1.0 x 1.0 m3): vertical flow vegetated bed over horizontal flow sand bed, were operated under an average temperature of 24 degrees C. Pig farm wastewater with COD and NH4-N concentration of 1034 and 448 mg/L in average was fed to the system at different HLR from 2 to 8 cm/day. The performance of the system when operated with a vertical flow bed followed by a horizontal flow bed or vice versa did not show a significant difference but under high HLR, nitrogen removal efficiencies were clearly reduced. Nitrobacter and Nitrosomonas were found in a large number in vertical flow beds and the same for denitrifier bacteria in a horizontal flow beds. Removal rate constants for nitrification (kNH4+ -N) were 0.0413 m/d for H-Vmode and 0.0339 m/d for V-H mode. Removal rate constants for denitrification (kNOx-N) were 0.0979 m/d for H-Vmode and 0.0399 m/d for V-H mode, respectively. PMID- 16042243 TI - Constructed wetland attenuation of nitrogen exported in subsurface drainage from irrigated and rain-fed dairy pastures. AB - Nitrogen removal performance is reported for constructed wetlands treating subsurface drainage from irrigated and rain-fed dairy pastures in North Island, New Zealand. Flow-proportional sampling of inflow and outflow concentrations were combined with continuous flow records to calculate mass balances for the wetlands. Drainage flows from the irrigated catchment were 2.5-4 fold higher and N exports up to 5 fold higher per unit area than for the rain-fed catchment. Hydraulic and associated N loadings to the wetlands were highly pulsed, associated with rainfall, soil water status, and irrigation events. Transient pulses of organic nitrogen were an important form of N loss from the rain-fed landscape in the first year, and were very effectively removed in the wetland (> 90%). Median nitrate concentrations of approximately 10 g m(-3) in the drainage inflows were reduced by 15-67% during passage through the wetlands and annual nitrate-N loads by 16-61% (38-31 7 g N m(-2)y(-1)). Generation in the wetlands of net ammoniacal-N and organic-N (irrigated site) partially negated reduction in nitrate-N loads. The results show that constructed wetlands comprising 1-2% of catchment area can provide moderate reductions in TN export via pastoral drainage, but performance is markedly influenced by variations in seasonal loading and establishment/maturation factors. PMID- 16042244 TI - Removal of nitrogen in different wetland filter materials: use of stable nitrogen isotopes to determine factors controlling denitrification and DNRA. AB - Laboratory incubations with varying O2 and NO3 concentrations were performed with a range of filter materials used in constructed wetlands (CWs). The study included material sampled from functioning CWs as well as raw materials subjected to laboratory pre-incubation. 15N-tracer techniques were used to assess the rates of denitrification versus dissimilatory nitrate reduction to ammonium (DNRA), and the relative role of nitrification versus denitrification in producing N2O. The N2O/(N2 + N2O) product ratio was assessed for the different materials. Sand, shell sand, and peat sustained high rates of denitrification. Raw light-weight aggregates (LWA) had a very low rate, while in LWA sampled from a functioning CW, the rate was similar to the one found in the other materials. The N2O/(N2 + N2O) ratio was very low for sand, shell sand and LWA from functioning CWs, but very high for raw LWA. The ratio was intermediate but variable for peat. The N2O produced by nitrification accounted for a significant percentage of the N2O accumulated during the incubation, but was dependent on the initial oxygen concentration. DNRA was significant only for shell sand taken from a functioning CW, suggesting that the establishment of active DNRA is a slower process than the establishment of a denitrifying flora. PMID- 16042245 TI - Treatment of dirty water from dairy farms using a soil-based batch recirculation system. AB - "Dirty water", a wastewater produced on dairy farms, is typically disposed of by application to land with no prior treatment. Pollution can occur if the dirty water reaches a watercourse following an inadequate period of retention in the soil. This paper describes experiments using a novel, soil-based batch recirculation system for pre-treating dirty water prior to land application. Three polythene-lined, vegetated soil-based treatment planes (23 m long, 1 m wide, 0.25 m deep) were constructed. Each treatment plane was supplied with approximately 1 m3 of dirty water which was recirculated until a clear treatment pattern had emerged. Five batches were treated over a six-month period. The soil based treatment system could typically be expected to achieve a 90% removal of key pollutants in approximately two weeks for BODs and NH4-N, and three weeks for MRP and total solids. An exponential trendline gave a good fit to the treatment curves for BOD5, NH4-N and MRP after the first day or two of batch treatment. The data for total solids removal were more variable. Treatment rates were sustained throughout the five runs for BOD5 and NH4-N, indicating no apparent effect of seasonal weather on the treatment process. The apparent progressive slowing of the MRP removal rate throughout the treatment of the five batches may have implications for the sustainable use of this technology for phosphorus control. PMID- 16042246 TI - The performance of vertical flow constructed wetland systems with special reference to the significance of oxygen transfer and hydraulic loading rates. AB - The paper reviews the development of the vertical flow (VF) reed beds/constructed wetlands over the past 20 years. The performance of VF systems (and their use within hybrid systems) is analysed by reference to a number of brief case studies. The oxygen transfer rate (OTR) achieved is absolutely critical to the sizing of the systems. The author reviews the reported OTRs and comments on the existing design equations proposed for calculation of the area of beds. The 1st generation of VF systems used a set of parallel beds that were dosed one at a time in rotation and then rested for a period of days because there was considerable concern (based on early experience) that they would become clogged. In the past 10 years a number of new designs of 2nd generation VF beds have been built which make use of a single bed and hence operate without any resting periods. The hydraulic loading rate and the selection of the bed media, which are critical to the design and hence successful operation of these 2nd generation compact VF beds, are described. It is now possible to produce a very high quality of effluent from VF beds alone sized at 2 m2/pe when treating domestic sewage. PMID- 16042247 TI - Influence of surface layer on hydrology and biology of gravel bed vertical flow constructed wetlands. AB - In France, gravel vertical flow constructed wetlands (gVFCWs) were adapted to treat raw wastewater, which led to important accumulations of matter in filters (organic and mineral). To prevent clogging, large gravel sizes were employed (O 2 6 mm). The aim of this paper was to present the influences of matter accumulation on the hydraulic and biological behaviour of the system. A one-year survey of accumulated matter content and potential respiration activities was completed in three gVFCWs (operating for 3, 4 and 8 years). Cores were sampled into filters. Results showed a vertical stratification of accumulated matter and respiration rates. Dry accumulated matter quantities ranged from 20 kg m(-2) (3 and 4 years operating) to 80 kg m(-2) in the oldest plant (8 years). Potential respiration was larger in the oldest plant (75g O2m(-2)h(-1)) than in the most recent one (15g O2m(-2)h(-1)). Accumulated matter seemed to play a role both on the water retention (enhancing initial percolation time by 5 times) and biological profile (enhancing microfauna's diversity). Contrary to what is generally proposed in the literature, accumulated matter in French gVFCWs seemed to provide better treatment efficiency without leading to surface clogging compared to systems using sand. PMID- 16042248 TI - Sludge reed bed facilities: operation and problems. AB - Short operating periods and problems with dewatering efficiency, vegetation, mineralisation and odour are primarily caused by incorrect construction of the filter, poor capillary connections, an inadequate number of basins, insufficient basin area and overloading during commissioning and during subsequent operation. Dimensioning should be based on the sludge quality including the dewatering qualities and not solely on calculations of the sludge production. Loading after planting depends on the development level of the vegetation. The loading program should ensure that reed establishment is not impeded and should prevent the sludge residue from growing so quickly horizontally and vertically that the reeds cannot colonise the sludge residue. Overloading results in an anaerobic sludge residue with ensuing methane production. Typically, a sludge reed bed facility with a loading period of maximum 5 days, must be built with 10 basins to permit a rest phase of about 40 days. Facilities with 8 basins, where it is possible to load 1 basin for 7 days, will be able to have a 7-week rest phase before the first basin is loaded again. Facilities with for example 8-13 basins and loading period of between 4 and 10 days are able to achieve rest periods of up to 2 to 2(1/2) months, which results in the optimal evapotranspiration and minerilization. The conditions and thus the possibilities vary depending upon the type of sludge. PMID- 16042249 TI - Mineralisation of hazardous organic compounds in a sludge reed bed and sludge storage. AB - Linear alkylbenzene sulphonates (LAS) and nonylphenolethoxylates (NPE) may be detrimental to the environment if spread in inappropriately large concentrations. Mineralisation of LAS and NPE in mesophilic digested sludge was observed during a 9-month monitoring programme where three separate treatment methods were investigated. Sludge was treated in a sludge reed bed under aerobic conditions, by storage in a container under anaerobic conditions, and by storage in a sludge pile turned over mechanically at intervals to improve the oxygen influx. Treatment in a sludge reed bed was shown to be effective. Mineralisation of 98% of LAS and 93% of NPE was observed. Only limited mineralisation occurred on the surface of the sludge stored in a container. A reduction of LAS and NPE of 90% and 43%, respectively, was observed in the sludge which was stored in a pile and frequently turned. PMID- 16042250 TI - Treatment of septage in constructed wetlands in tropical climate: lessons learnt from seven years of operation. AB - In tropical regions, where most of the developing countries are located, septic tanks and other onsite sanitation systems are the predominant form of storage and pre-treatment of excreta and wastewater, generating septage and other types of sludges. The septage is disposed of untreated, mainly due to lack of affordable treatment options. This study presents lessons that have been learned from the operation of pilotscale constructed wetlands (CWs) for septage treatment since 1997. The experiments have been conducted by using three CW units planted with narrow-leave cattails (Typha augustifolia) and operating in a vertical-flow mode. Based on the experimental results, it can be suggested that the optimum solids loading rate be 250 kg TS/m2 yr and 6-day percolate impoundment. At these operational conditions, the removal efficiencies of CW units treating septage at the range of 80-96% for COD, TS and TKN were achieved. The biosolid accumulated on the CW units to a depth of 80 cm has never been removed during 7 years of operation, but bed permeability remained unimpaired. The biosolid contains viable helminth eggs below critical limit of sludge quality standards for agricultural use. Subject to local conditions, the suggested operational criteria should be reassessed at the full-scale implementation. PMID- 16042251 TI - Factors affecting the longevity of sub-surface horizontal flow systems operating as tertiary treatment for sewage effluent. AB - Compared with other forms of domestic wastewater treatment, reed beds require very little in the way of operational and maintenance input. This is even more markedly the case with tertiary treatment systems (than with secondary treatment systems) where the necessary maintenance amounts to only a few days/year. Unfortunately, in practice this frequently results in them receiving little or no attention at all. Tertiary treatment reed beds are not a "fit and forget" solution but they are often treated this way because they are very forgiving and abuse-tolerant. After a number of years a few of these tertiary reed beds have deteriorated to an extent whereby they are close to failing to comply with the regulator's requirements. Severn Trent Water Ltd has recognised this situation and has committed a budget for a programme of reed bed maintenance. This work has started with a survey of the condition of their beds and particularly to determine the common problems and hence the maintenance regime required. Five main problems occurred either singly or in combination namely: (a) sludge deposition; (b) above surface flooding (partially caused by (a), (c), and (d)); (c) inlet flow distributor problems/clogging; (d) outlet collector problems/incorrect level; (e) weed infestation. PMID- 16042252 TI - The effect of a vertical flow filter bed on a hybrid constructed wetland system. AB - Data from 18 sampling wells in Kodijarve horizontal subsurface flow (HSSF) constructed wetland (CW) (South Estonia) is presented and differences in purification efficiencies inside the HSSF CW are calculated. Temporarily anaerobic conditions in the Kodijarve HSSF system did not allow efficient removal of BOD7, NH4-N, Ntot and Ptot. In 2002 a vertical subsurface flow filter was constructed to enhance aeration. The design of the system was based simply on the oxygen demand of the wastewater and on the aeration potential of vertical flow wetlands. The vertical flow system has shown satisfactory results. The purification efficiency of BOD7 in the Kodijarve CW has improved significantly and there has been a slight increase in purification efficiencies of NH4-N and Ntot. On the ohther hand, the removal efficiency of Ptot has decreased significantly. Although, the mass loading rates have increased, mass removal rates of all four parameters have improved significantly. Nevertheless, optimization of the constructed wetland system is essential in order to meet effluent standards during wintertime. PMID- 16042253 TI - Vertical flow constructed wetlands for municipal wastewater and septage treatment in French rural area. AB - This paper presents the purification performance of 20 wastewater treatment plants with vertical reed bed filters (Macrophyltres), built between 1998 and 2003 by SAS Voisin, for communities of between 150 and 1400 PE. The first stage vertical reed bed (directly fed with raw wastewater by intermittent feeding) achieved high removal of SS, BOD and COD (mean respectively 96%, 98%, 92%). The second stage permitted compliance easily with effluent standards (SS < 15 mg/l, BOD < 15 mg/l, COD < 90 mg/l and mean TKN < 10 mg/l). Performance was not significantly influenced by variations of organic and hydraulic load, nor by seasonal variations. Rigorous operation and maintenance were required to obtain optimal performances. Another application of vertical reed beds is the treatment of septage (sludge from individual septic tanks). The results obtained on two sites operating for 2 and 3 years are presented. The first site achieved complete treatment of septage (solid and liquid fraction), the second permitted a pre treatment for co-treatment of percolate with wastewater. PMID- 16042254 TI - Treatments of oil-refinery and steel-mill wastewaters by mesocosm constructed wetland systems. AB - In this study, two types of industrial wastewater, oil-refining and steel milling, were selected for investigating their feasibility of treatment by mesocosm constructed wetland systems. The secondly treated effluents from the wastewater treatment plants were directly discharged into the systems controlled at different flow rates. Three wetland mesocosms were installed in the two industries: mesocosms A and B were in the oil refinery, and mesocosm C was in the steel mill. The substratum media used in wetland systems were sand (mesocosm A) and gravel (mesocosms B and C), while the vegetation types selected were reeds (mesocosms A and B) and mixed species of reeds and cattails (mesocosm C). The flow regimes were controlled as free water surface (FWS) and subsurface flow (SSF) for the sand- and gravel-beds, respectively. According to the experimental results, we found that the system treating oil-refining wastewater performed better than that treating steel-milling wastewater learned by comparing the removal efficiencies of COD, total N and total P. In addition, it was found that for oil-refining wastewater treatments, the SSF wetland system (mesocosm B) performed better than FWS (mesocosm A) wetland system when comparing both of their removal of pollutants and growth of vegetation. Besides, the effluents from these two industrial wetland treatment systems might be reclaimed and reused for boiler water, cooling, cleaning and miscellaneous purposes in industries. Further treatments are required if the constructed wetland effluents are thought about being reused for processing in industries. PMID- 16042255 TI - BTEX degradation in a cold-climate wetland system. AB - A pilot-scale subsurface vertical-flow wetland system was constructed at the former BP Refinery in Casper, Wyoming in order to determine benzene, toluene, ethylbenzene and xylene (BTEX) degradation rates in a cold-climate application. The pilot system, consisting of 4 cells, each dosed at a nominal flow rate of 5.4 cubic metres per day, was operated between August and December 2002. The pilot tested the effects of wetland mulch and aeration on system performance. Areal rate constants (kA values) were calculated based on an assumed three tanks in series (3TIS). The presence of wetland sod and aeration both improved treatment performance. Mean kA values were 244 m/yr for cells without sod or aeration, and improved to 356 m/yr for cells with sod and aeration. Based on the results of the pilot system, a full-scale wetland system (capable of operating at 6,000 m3/day) was started up in May 2003. The full-scale system achieved permit compliance within one week of startup, but is currently being loaded at only 45% of the design hydraulic load, and 15% of the design BTEX mass load, resulting in a mean kA value of approximately 350 m/yr. PMID- 16042256 TI - Potential of a tropical subsurface constructed wetland to remove phenol from pre treated pulp and papermill wastewater. AB - The capacity of a pilot project subsurface flow constructed wetland in the tropics to remove phenol from pre-treated pulp and paper mill wastewater was studied under varying hydraulic retention times (HRT) with batch loading. Initial 15 months results indicate that removal efficiencies for phenol were variable but on average reached 60% at 5-day HRT and 77% at 3-day HRT. It was thought that the longer retention time might have caused oxygen and nutrient deficiencies, which may have reduced removal performance. Although phenol was sometimes not detectable in the wetland outflow, on average values over the experimental period did not meet set national guidelines. In the ongoing study, the impact of varying hydraulic retention time and/or loading rate on the removal of phenols will be evaluated and the main removal process established. PMID- 16042257 TI - Processes in vertical-flow reed beds: nitrification, oxygen transfer and soil clogging. AB - Vertical-flow reed beds (VF) with intermittent feeding are extremely reliable regarding aerobic processes. For a save operation with high nitrification rates and without soil clogging it is essential to preserve aerobic conditions in the filter. The challenge is to keep aerobic conditions in the filter without oversizing the system (economical aspects). It is very difficult to determine the current oxygen content in the filters because it ultimately results from complex interactions of a large number of different influencing parameters such as loading rate, degree of clogging, temperature, and hydraulic behaviour of the reed bed. To gain better knowledge of this complex system, different tests and examinations were carried out over several years. Focusing on the questions of identification and the description of conversion and transport processes (water/gas), a full-scale treatment plant under clogged and non-clogged conditions was investigated in detail. Additionally soil column test were carried out. The results make it possible to describe some of the processes and their interactions in the filter body. Recommendations for a safe and controlled operation can be derived. PMID- 16042258 TI - Root stimulated nitrogen removal: only a local effect or important for water treatment? AB - Plants in constructed wetlands serve as carriers for attached microbial growth. They mainly transfer oxygen and release exsudates to the root zone. In consequence of this an area around the roots, called the rhizosphere exists, in which bacteria are stimulated by root growth. The goals were to ascertain whether stimulating the microbial cenosis only has a local effect on the rhizoplane, and to establish the importance of this stimulation for the water purification process in the root zone. Observations were carried out in a laboratory batch reactor filled with sand and planted with reeds (Phragmites australis). A small section was separated with gauze to avoid root growth outside this zone. The reactor was incubated with an artificial waste water containing a high concentration of ammonium. Samples were taken at intervals of 10 mm away from the gauze. The chemical and physical conditions and enzyme activities in soil sections at different distances from the roots affecting the efficiency of nitrogen removal were characterized. An influence was detectable by several parameters up to a specific root distance. Indirect parameters such as the total bacterial number and the DNA amount seem to be affected up to a distance of 50 mm from the root whereas the oxygen amount and DOC are unaffected at a distance exceeding 20-30 mm. This is an initial indication that improved nitrogen removal is also possible in the wider root surroundings. In view of the average root-to root distance of 35 mm, the root-influenced area could therefore be expanded to the whole rooted zone in a constructed wetland. The influence on bacteria by roots is not just a local effect but may also play an important role in the whole purification process. PMID- 16042259 TI - Apatite as an interesting seed to remove phosphorus from wastewater in constructed wetlands. AB - Intensive use of phosphates has resulted in high P levels in surface waters and therefore eutrophication problems. Over the last decade many studies have revealed the advantage of using specific materials with efficient phosphorus retention capacities. Recent studies state that Ca materials are of particular interest for long-term retention of P, but can induce negative effects. To improve P retention and avoid negative counter-effects we tested the potential of natural apatites. Apatite sorption was evaluated using batch and open reactor experiments. Batch experiments identify sorption mechanisms and the influence of the ionic characteristics of the solution; open reactor experiments evaluate sorption capacities in relation to the ionic composition of the solution and biomass development. In parallel, observation of the material by electron microscopy was used to give more precision information about the mechanisms involved. This work reveals the strong chemical affinity between apatites and phosphorus. Compared to other calcareous materials apatite is better able to maintain low outlet P levels. After more than 550 days feeding, sorption was still present and low P outlet levels were still being obtained when sufficient contact time and calcium content in the solution were ensured. This work demonstrates the advantages of using apatites for phosphorus removal in constructed wetlands. The behaviour of apatite in phosphorus retention is explained and its suitability for use in such extensive systems defined. PMID- 16042260 TI - Retention of Escherichia coli in municipal sewage by means of planted soil filters in two-stage pilot plant systems. AB - Different types of constructed wetlands in a pilot-plant system were fed with identical municipal waste water to compare the sanitisation process in two-stage systems. With combinations of a vertical and a horizontal flow filter an E. coli reduction of 5 log10 with an E. coli influent concentration of 10(7) MPN/100 ml was achieved. Using different filter materials in each stage the total performance of the two-stage system was independent from the sequence of these materials. However, using coarser filter material in the first stage makes the filter less prone to clogging and is thus the preferential option with regard to operational reliability. PMID- 16042261 TI - The role of plant uptake on the removal of organic matter and nutrients in subsurface flow constructed wetlands: a simulation study. AB - Plants in constructed wetlands have several functions related to the treatment processes. It is generally agreed that nutrient uptake is a minor factor in constructed wetlands treating wastewater compared to the loadings applied. For low loaded systems plant uptake can contribute a significant amount to nutrient removal. The contribution of plant uptake is simulated for different qualities of water to be treated using the multi-component reactive transport module CW2D. CW2D is able to describe the biochemical elimination and transformation processes for organic matter, nitrogen and phosphorus in subsurface flow constructed wetlands. The model for plant uptake implemented describes nutrient uptake coupled to water uptake. Literature values are used to calculate potential water and nutrient uptake rates. For a constructed wetland treating municipal wastewater a potential nutrient uptake of about 1.9% of the influent nitrogen and phosphorus load can be expected. For lower loaded systems the potential uptake is significantly higher, e.g. 46% of the nitrogen load for treatment of greywater. The potential uptake rates could only be simulated for high loaded systems i.e. constructed wetlands treating wastewater. For low loaded systems the nutrient concentrations in the liquid phase were too low to simulate the potential uptake rates using the implemented model for plant uptake. PMID- 16042262 TI - Simulation of a subsurface vertical flow constructed wetland for CSO treatment. AB - Constructed wetlands (CWs) have proved to be a highly effective measure to reduce the ecological impact of combined sewer overflows (CSOs) on receiving waters. Due to the stochastic nature of the loading regime and the multitude of environmental influences, assessment of the performance of such plants requires detailed mathematical modelling. A multi-component reactive transport module (CW2D) was applied to simulate the flow, transport and degradation processes occurring in a CW for CSO treatment. CW2D was originally developed to simulate the treatment of municipal wastewater in subsurface flow CWs. Loading and operational conditions in CSO treatment differ fundamentally from the conditions occurring for wastewater treatment. Despite these differences, first results from the simulation of lab-scale experiments show, that the model is generally applicable to this type of plant. Modelling of adsorption, degradation processes, and influent fractionation, however, require further research. PMID- 16042263 TI - In-situ monitoring of microbial biomass in wetland mesocosms. AB - The objective of the present investigation has been to combine tracer principles and a hydrolytic microbial activity assay using fluorescein diacetate to monitor changes in microbial biomass within subsurface flow wetland mesocosms. The mesocosm hydrolytic activity was referenced to activated sludge concentrations treating a typical domestic wastewater at full scale. Microbial biomass activity levels within four laboratory wetland mesocosms treating a synthetic domestic wastewater were routinely monitored over a 21-week period of plant growth and rhizosphere development. Although above ground plant mass and tracer dispersion numbers suggested progressive root zone development, plant growth did not result in any measurable enhancement in microbial activity when compared to a mesocosm operating without plants. Dispersion numbers also suggested a reduction in the mass transport kinetics in these planted mesocosms. In-situ biomass monitoring enabled the assessment of a characteristic response in terms of the steady-state food to microorganism (F/M) ratio that was observed in mesocosms receiving both low and high organic loading. Wetland treatment performance is sensitive to the degree to which bed volume is exploited in terms of wastewater flow to regions of bioactivity. The in-situ reactive tracer technique for mesocosm biomass monitoring provided an assessment of the collective substratum and rhizosphere microbial biomass in direct contact with wastewater contaminants. Thus, in-situ biomass monitoring has application in further understanding of plant function and strategies for plant implementation in wetland research and development. PMID- 16042264 TI - Impact of operational maintenance on the asset life of storm reed beds. AB - This paper reviews the operation of storm reed beds to determine whether the current system of operational maintenance is contributing to premature process failures and if not, to identify other factors of importance. Twelve storm reed beds of the horizontal subsurface flow type, at seven locations in the South Warwickshire area of the United Kingdom, were surveyed. Each survey consisted of a site visit, an interview with the operators in charge and an assessment of the treatment performance based on routine monitoring data. Although some sites suffered from varying degrees of sludge accumulation, surface blinding and/or weed growth, all effluent concentrations remained far below the consent levels. Thorough operational maintenance on a reed bed is proven to be important for the asset life. However, there are other factors or features of a reed bed that play a more pivotal role in premature process failure such as the lack of pretreatment and a premature operation of the storm overflow. PMID- 16042265 TI - Treatment of gully pot liquor containing heavy metals with constructed wetlands in Scotland. AB - The aim of this investigation was to assess the treatment efficiencies for gully pot liquor of 12 experimental vertical-flow constructed wetland filters containing Phragmites australis (Cav.) Trin. ex Steud. (Common Reed) and filter media of different adsorption capacities in a cold climate. Seven of the twelve filters received inflow water spiked with heavy metals. For one year, hydrated copper nitrate and hydrated nickel nitrate were added to sieved gully pot liquor to simulate contaminated primary treated storm water runoff. The inflow concentrations for dissolved copper, nickel and nitrate-nitrogen were approximately 1.0, 1.0 and 1.45 mg/l, respectively, which represent mean loading rates of 0.063 g/m2/d for Filters 2 and 7 to 11, and 0.115 g/m2/d for Filter 12. For these filters receiving metals, an obvious breakthrough of dissolved nickel was recorded after road gritting and salting during winter. Sodium chloride was responsible for nickel leaching. Reductions of copper, nickel, biochemical oxygen demand and suspended solids were frequently insufficient compared to international secondary wastewater treatment standards. Moreover, the overall filtration performance for all filters was similar. PMID- 16042266 TI - The effect of the scale of horizontal subsurface flow constructed wetlands on flow and transport parameters. AB - Horizontal subsurface flow constructed wetlands have proven their efficiency in treating wastewater and removing the pollutants of concern. Treatment efficiency depends on the wastewater residence time, which is a function of the hydraulic loading and the physical conditions of the constructed filter system, which can be described with effective parameters such as: hydraulic conductivity, porosity, dispersivity etc. Because spatial variability is often scale dependent, these effective parameters may be affected by the scale of the system being studied. In this paper the results of tracer experiments in constructed filters using saturated horizontal flow at three scales (small and medium lab scales and full scale system) using the same filter media is reported. Light-weight aggregate (filter media termed Filtralite-P) was used at all scales. Increasing the scale was associated with increasing dispersivity, meanwhile hydraulic conductivity experienced dramatic reduction and variation by increasing the examined scale. Observed changes in the hydraulic parameters indicate that heterogeneity at different scales should be taken into account when the performance of LWA filters are evaluated from small-scale experiments. PMID- 16042267 TI - Phosphorus removal in constructed wetlands: can suitable alternative media be identified? AB - Removal of phosphorous in constructed wetlands is limited by the capacity of the media to adsorb, bind or precipitate the incoming P. To enhance P removal and the life span of constructed wetlands the approach might be to use natural sands rich in calcium or iron, to use an alternative 'artificial' medium with high P-binding capacity, or to establish external P-binding filters after the wetland. Our studies focused on the evaluation of calcium-rich materials potentially useful as P-binding media. The materials tested included calcite products, natural sands and seashells. Tests included assessment of physical and chemical properties of the materials, extractions in P-spiked water at different P concentrations to determine P-binding equilibrium isotherms, and column experiments. In addition, full-scale tests were performed with calcite in an external filter. The result showed that equilibrium isotherm is an indicator of the potential P-sorption capacity of the media, although the value is of limited application for the determination of the binding capacity in full-scale systems. The columns showed that the materials do bind phosphorus. However, the binding capacities are still insufficient for the establishment of external P-removal filter; the volumes of the filters would be too large to be of practical use. PMID- 16042268 TI - Enhancing phosphorus removal in constructed wetlands with ochre from mine drainage treatment. AB - No single end-use has yet been identified that is capable of consuming the projected production of ochre (mainly iron (III) oxides) from mine drainage treatment. However, the high sorption capacity of ochre for phosphorus (up to 26 mg kg(-1)) means that it could be used in constructed wetlands to enhance phosphorus removal. Laboratory batch experiments showed that coarse-grained ochre removes 90% of all phosphorus forms from sewage effluent after 15 minutes of shaking. From a larger-scale experiment, it is estimated that constructed wetlands with an ochre substrate should remove phosphorus from sewage effluent for up to 200-300 years. The suitability of ochre for phosphorus removal is being investigated at the field scale in a wastewater constructed wetland (175 m2 area) in Berwickshire, UK. The hydraulic and treatment performance of the wetland were monitored for 15 months prior to installation at the inlet in November 2003 of a tank containing approximately 1200 kg ochre. Results so far show that improved hydraulic design is required for ochre to increase the mean phosphorus removal efficiency of the system (27 +/- 28%), but potentially toxic metals (Al, Cd, Cr, Cu, Fe, Ni, Pb, Zn) have not been released from the ochre into the wetland outflow. PMID- 16042269 TI - Optimization of low-cost phosphorus removal from wastewater using co-treatments with constructed wetlands. AB - Eighteen wastewater treatment systems were operated for one year to investigate phosphorus (P) removal. Systems paired co-treatment reactors containing iron or calcium drinking water treatment residuals with vertical-flow constructed wetland mesocosms planted with Schoenoplectus tabernaemontani (K.C. Gmel.) Palla. For secondary municipal wastewater, soluble reactive P (SRP) concentrations were reduced from 0.70 to 0.03 mg L(-1) (95%) or 0.01 mg L(-1) (98%) by systems with the calcium or iron co-treatments, respectively (compared to 0.09 mg L(-1) or 87% by controls). Total P (TP) concentrations were reduced from 1.00 to 0.07 mg L(-1) (93%) and 0.05 mg L(-1) (95%) by the same treatments (compared to 0.16 mg L(-1) or 84% by controls). For anaerobically digested dairy wastewater, SRP was reduced from 7.68 to 6.43 mg L(-1) (16%) or 5.95 mg L(-1) (22%) by the systems with calcium or iron, respectively (compared to 7.37 mg L(-1) or 4% by controls). For this wastewater, the TP was reduced from 48.5 to 22.5 mg L(-1) (53%) and 22.7 mg L(-1) (53%) by the same treatments (compared to 24.1 mg L(-1) or 50% by controls) but performance improved substantially with a design modification tested. PMID- 16042270 TI - Wetland to pond treatment gradients. AB - This paper presents information on the hydraulic efficiency of wetlands and ponds, and examines the differences and similarities between degrees of mixing, short-circuiting and dead zones. It reports the pollutant removal performance of over 50 wetlands and ponds over the range of macrophyte cover from 100% (densely vegetated wetlands) to 0% (ponds). Not surprisingly, there exist performance continua, characterized by a smooth transition from fully vegetated to unvegetated systems. Factors influencing the choice of ponds, wetlands or combinations are examined. Wetlands are more efficient, and can produce lower TSS. Both wetlands and ponds can provide BOD, FC and ammonia removal, with ponds being better choices at high loading rates. PMID- 16042271 TI - Exceeding tertiary standards with a pond/reed bed system in Norway. AB - At Vidarasen in Norway sewage from a community consisting of 160 people, including a dairy, a food processing workshop, a bakery and a laundry is treated using a pond/reed bed system. The system consists of sludge settlement, pre treatment surface/vertical-flow constructed wetlands, a 5 m deep enhanced facultative pond, three stabilization ponds, a planted sand filter and finally two horizontal-flow constructed wetlands filled with lightweight aggregate (Filtralite-P). The enhanced facultative pond and the primary stabilization pond are equipped with Flowform-cascades, which provide year-round aeration, rhythmical treatment and mixing of wastewater in the ponds. Treatment performance during the first five years has been high and unaffected by harsh winter conditions. Average phosphorus discharge from the system is 0.25 mg/l with total nitrogen 4 mg/l, total organic carbon (TOC) 5 mg/l and thermo-tolerant coliforms < 100/100 ml. The system is ecologically diverse and supports abundant populations of higher aquatic life such as ducks, amphibians and carp. PMID- 16042272 TI - Sediment deposition in constructed wetland ponds with emergent vegetation: laboratory study and mathematical model. AB - Retention of suspended particles by settling is among the main physical treatment processes in constructed wetland ponds. Laboratory experiments were conducted to study the transport and deposition of suspended particles in the slow, near stagnant flows typical of constructed wetland ponds with emergent vegetation. The presence of stems was found to create a velocity field which is much more uniform than its counterpart without vegetation. This property was used to obtain an approximate mathematical model, for which an analytical solution could be given to describe sediment transport and deposition. The deposition rates predicted by this formula were compared to the data from the above-mentioned laboratory experiments and found to agree closely. PMID- 16042273 TI - Influence of biofilm on removal of surrogate faecal microbes in a constructed wetland and maturation pond. AB - The effect of biofilm on the attenuation of pathogen-sized particles from wastewater was compared for biofilms cultivated in a surface flow constructed wetland (SFW) and maturation pond (MP) The fate of fluorescently labelled microspheres (FLM) as surrogates for viruses (0.1 microm), bacteria (1 microm) and parasitic protozoa (4.5 microm dia) was investigated in microcosms in the presence or absence of biofilms. Rates of FLM removal from suspension were higher in the presence of biofilms for all particle sizes (kd 0.02-0.11 h(-1)) in MP and SFW microcosms with removal efficiency related to particle size and biofilm thickness and structure. Greater removal of 0.1 microm (79-81%), 1 microm FLM (92 96%) and 4.5 microm FLM (up to 98%) from suspension were found for microcosms containing thicker (autotrophic) biofilms grown in the MP or open water zone of the SFW. Lower removal of 43% (0.1 microm), 59% (1 microm) and 84% (4.5 microm) occurred in microcosms containing thinner heterotrophic biofilms from SFW vegetated zones. Providing surfaces for attachment of photosynthetic biofilms offers potential to enhance pathogen removal in open water systems. In vegetated systems, linkage to more oxic openwater zones may allow thicker and 'stickier' epiphytic biofilms to develop, improving pathogen interception and removal. PMID- 16042274 TI - Hydroxylated PCB metabolites in nonhatched fulmar eggs from the Faroe Islands. AB - Thirty-six polychlorinated biphenylols (OH-PCBs) congeners were characterized in Fulmar (Fulmarus glacialis) eggs collected from the Faroe Islands. The seven most abundant congeners were quantified in 19 samples, and the sigmaOH-PCB concentrations ranged between 0.92 and 4.0 ng g(-1) fresh weight (f.w.). These eggs constitute a part of the traditional diet for at least a part of the population on the Faroe Islands and may contribute to the high levels of these contaminants found in the blood of pregnant Faroese women. Because the metabolites are present in the nonhatched fulmar egg, it is concluded that the OH PCBs are transferred to the egg before laying. High levels, 3300-18,000 ng g(-1) l.w., of sigmapolychlorinated biphenyls (PCB) were determined in the fulmar eggs, which are a considerable source for human exposure. The high PCB levels are a source for metabolic formation of hydroxylated PCBs. PMID- 16042275 TI - Recent changes in trophic state of the Baltic Sea along SW coast of Finland. AB - Drawing reliable conclusions on changes in the trophic state of various subareas of the Baltic Sea is problematic, in large part because the monitoring of productivity parameters exhibiting high natural variability is based on sparse sampling. This emphasizes the importance of long-term data sets. Here we present a 30-year chlorophyll alpha data set from the western Gulf of Finland. The trophic state of the study area showed an increasing trend in the 1970s and 1980s manifested mainly as strengthened vernal blooms. This trend did not continue in the 1990s, and the seasonal phytoplankton biomass maxima since then has begun to show some bias toward the late summer. The changes in seasonal maxima of phytoplankton blooms probably reflect i) decreased availability of N suppressing the magnitude of the vernal bloom, which increases the P reserves for the summer, and ii) enhanced internal P loading, which further increases the summer P reserves. PMID- 16042276 TI - Abrupt biological response to hydrologic and land-use changes in Lake Apopka, Florida, USA. AB - Lake Apopka is a shallow, hypereutrophic lake in north-central Florida that experienced an abrupt shift in primary producer community structure (PPCS) in 1947. The PPCS shift was so abrupt anecdotal accounts report that dominant, submersed aquatic vegetation was uprooted by a hurricane in 1947 and replaced by phytoplankton within weeks. Here we propose two hypotheses to explain the sudden shift to phytoplankton. First, hydrologic modification of the drainage basin in the late 1800s lowered the lake level ca. 1.0 m, allowing the ecosystem to accommodate moderate, anthropogenic nutrient enrichment through enhanced production in the macrophyte community. Second, additional hydrologic changes and large-scale agricultural development of floodplain wetlands began in 1942 and altered the pattern and scale of phosphorus loading to the lake that triggered the rapid shift to phytoplankton dominance in 1947. Historic land-use changes and paleolimnological data on biological responses to nutrient loading support these hypotheses. PMID- 16042277 TI - Agricultural capacity and conservation in high biodiversity forest ecosystems. AB - Agricultural development is a leading cause of habitat destruction that increasingly threatens global biodiversity. To help understand the likelihood and implications of agricultural expansion in areas of high conservation importance, this article examines agricultural suitability in forested portions of biodiversity hotspots and tropical wilderness areas, regions with especially rich concentrations of species found nowhere else. The study employs geographic information system technology to examine suitability for six crop categories in selected conservation localities worldwide: those portions of regions containing high biodiversity, protected areas (e.g. national parks) within these regions, and 10-km bands around the protected areas that are dominated by forest. Analyses reveal low suitability for most crop categories under both commercial and subsistence scenarios, with a few exceptions. In most cases, adequate planning can enable the coexistence of agriculture and biodiversity without compromising either. PMID- 16042278 TI - Illicit crops and armed conflict as constraints on biodiversity conservation in the Andes region. AB - Coca, once grown for local consumption in the Andes, is now produced for external markets, often in areas with armed conflict. Internationally financed eradication campaigns force traffickers and growers to constantly relocate, making drug related activities a principal cause of forest loss. The impact on biodiversity is known only in general terms, and this article presents the first regional analysis to identify areas of special concern, using bird data as proxy. The aim of conserving all species may be significantly constrained in the Santa Marta and Perija mountains, Darien, some parts of the Central Andes in Colombia, and between the middle Maranon and middle Huallaga valleys in Peru. Solutions to the problem must address the root causes: international drug markets, long-lasting armed conflict, and lack of alternative income for the rural poor. PMID- 16042279 TI - Nest placement of the giant Amazon river turtle, Podocnemis expansa, in the Araguaia River, Goias State, Brazil. AB - The giant Amazon river turtle (Podocnemis expansa) nests on extensive sand bars on the margins and interior of the channel during the dry season. The high concentration of nests in specific points of certain beaches indicates that the selection of nest placement is not random but is related to some geological aspects, such as bar margin inclination and presence of a high, sandy platform. The presence of access channels to high platform points or ramp morphology are decisive factors in the choice of nesting areas. The eroded and escarped margins of the beaches hinder the Amazon river turtle arriving at the most suitable places for nesting. Through the years, changes in beach morphology can alter nest distribution. PMID- 16042280 TI - Conservation and development in Amazonian extractive reserves: the case of Alto Jurua. AB - Extractive reserves constitute an innovative approach to match conservation and development objectives, which were originally envisaged as part of a land struggle by forest dwellers in Brazil. In spite of the idea's popularity and the attempts to apply the concept to different tropical regions, there has been little analysis of the combined conservation and development performance of extractive reserve programs. We present a detailed analysis of deforestation and demographic and socioeconomic changes in Alto Jurua, the first extractive reserve created in Brazil in 1990. Forest cover has remained fairly stable. Population has declined slightly, with some internal displacements. The cash economy base has shifted from the original rubber production to a diversified portfolio of agriculture and livestock, and there has been a dramatic rise in nonagrarian income. We conclude that the Reserve represents a very dynamic setting with positive conservation and development outcomes during its first decade. PMID- 16042281 TI - The ecological and economic potential of carbon sequestration in forests: examples from South America. AB - Costs of reforestation projects determine their competitiveness with alternative measures to mitigate rising atmospheric CO2 concentrations. We quantify carbon sequestration in above-ground biomass and soils of plantation forests and secondary forests in two countries in South America-Ecuador and Argentina-and calculate costs of temporary carbon sequestration. Costs per temporary certified emission reduction unit vary between 0.1 and 2.7 USD Mg(-1) CO2 and mainly depend on opportunity costs, site suitability, discount rates, and certification costs. In Ecuador, secondary forests are a feasible and cost-efficient alternative, whereas in Argentina reforestation on highly suitable land is relatively cheap. Our results can be used to design cost-effective sink projects and to negotiate fair carbon prices for landowners. PMID- 16042282 TI - Geospatial indicators of emerging water stress: an application to Africa. AB - This study demonstrates the use of globally available Earth system science data sets for water assessment in otherwise information-poor regions of the world. Geospatial analysis at 8 km resolution shows that 64% of Africans rely on water resources that are limited and highly variable. Where available, river corridor flow is critical in augmenting local runoff, reducing impacts of climate variability, and improving access to freshwater. A significant fraction of cropland resides in Africa's driest regions, with 39% of the irrigation nonsustainable. Chronic overuse and water stress is high for 25% of the population with an additional 13% experiencing drought-related stress once each generation. Paradoxically, water stress for the vast majority of Africans typically remains low, reflecting poor water infrastructure and service, and low levels of use. Modest increases in water use could reduce constraints on economic development, pollution, and challenges to human health. Developing explicit geospatial indicators that link biogeophysical, socioeconomic, and engineering perspectives constitutes an important next step in global water assessment. PMID- 16042283 TI - An actor game on implementation of environmental quality standards for nitrogen in a Swedish agricultural catchment. AB - Despite political efforts, diffuse pollution from agriculture continues to be the single largest source of nitrogen (N) emissions into the aquatic environment in many countries and regions. This fact, and the recent enactment of a new Swedish environmental code, led to the design of a study targeted at the evaluation of new N pollution abatement strategies. An actor game was chosen as the key component of the study, with a focus on four major goals: to test the implementability of legally binding environmental quality standards for nitrate concentration in groundwater and N transport to the sea, to find sets of agriculturally feasible and cost-effective measures to decrease N loads, to investigate the possibilities for collective action through negotiated and institutionalized actor cooperation, and to investigate the role of mathematical modeling in environmental N management. Characteristics from the agriculturally dominated catchment of Genevadsan (224 km2) on the southwest coast of Sweden served as the playing field for the actor game. The most noteworthy result from the study was that it appeared to be possible to meet ambitious environmental N standards with less economically drastic measures than anticipated by most of the participants. The actor game was shown to be a good method for learning about the new Swedish environmental code and its application and for gaining deeper insight into the issues of N management. In addition, the actor game functioned as an arena for gaining a more thorough understanding of the views of different stakeholders. PMID- 16042284 TI - Government plans and farmers intentions: a study on forest land use planning in Vietnam. AB - In 1998, the Vietnamese National Assembly approved a Five Million Hectare Reforestation Program, (5MHRP) 1998-2010. It would increase forest cover by some 45%, use barren hills, produce wood, and generate socioeconomic development. It would, however, also put demands on Vietnamese planning. Experiences from a former development project and two case studies from a commune and village are used as a basis for analyzing the planning system. The study describes the objectives and strategies of government and farmers in one commune and one village and analyzes how planning data influence scenarios on future development. Official planning data do not reflect reality but are derived through negotiations. In the commune and village studied, it would be difficult for the 5MHRP to materialize, as most of the forest land, officially not yet used, is actually used for food production. The approach and method used by the study offers alternatives to current planning practices. PMID- 16042285 TI - Environmental threats to buried archaeological remains. AB - The last century's environmental pollution has created health problems, acidification of ground and lakes, and serious damage to our cultural heritage. Outdoor monuments suffer from this pollution, but so do buried archaeological remains. However, research on the deterioration of archaeological artifacts underground has so far been limited, and it is important to draw attention to this neglected field. This article presents results obtained at the Swedish National Heritage Board on the degradation of archaeological objects of bronze and iron and of bones from prehistoric graves, materials of which seem to be most affected by pollutants. The investigation methods, which were employed, are described. Other relevant studies are briefly reviewed. It is obvious that the deterioration rate of archaeological artifacts, especially of inorganic materials, has accelerated in recent years, and that this increased deterioration to a large part can be attributed to anthropogenic pollution. Regions that might be endangered are exemplified. PMID- 16042286 TI - Recent global warming: an artifact of a too-short temperature record? PMID- 16042287 TI - Globalization and soybean expansion into semiarid ecosystems of Argentina. PMID- 16042288 TI - Water management in megacities. PMID- 16042289 TI - Risk of extinction of a rare catfish of Andean groundwater and its priority for conservation. PMID- 16042290 TI - Summing up: a 10-y interdisciplinary research venture on sustainable water management. PMID- 16042291 TI - Determination and confirmation of five phenolic antioxidants in foods by LC/MS and GC/MS. AB - Identification and determination of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), nordihydroguaiaretic acid (NDGA), propyl gallate (PG) and tert-butylhydroquinone (TBHQ) by means of LC/MS and GC/MS were examined. These five phenolic antioxidants were detected as their pseudo-molecular ions [M-H]- by LC/MS using a Shim-pack FC-ODS column with drying gas. Moreover, BHA, BHT and TBHQ were detected based on their mass fragment ions by GC/MS. Decomposition of TBHQ, NDGA and PG during analysis could be prevented by the addition of L ascorbic acid (AsA) to the extraction solvent. All five antioxidants were extracted from nikuman, olive oils, peanut butter, pasta sauce and chewing gum with a mixture of acetonitrile-2-propanol-ethanol (2:1:1) containing 0.1% AsA (AsA mixture), which had been cooled in a freezer and filtered. One part filtrate and 5 parts water were mixed and placed on a Mega-Bond Elut C18 cartridge, except in the case of chewing gum. Lipids in foods were removed on a C18 cartridge by washing with 5 mL of 5% acetic acid, and antioxidants were eluted with 5 mL of AsA mixture. The antioxidants spiked into nikuman, olive oil, peanut butter, pasta sauce and chewing gum were successfully identified and their concentrations determined by LC/MS, and GC/MS with good recoveries. PMID- 16042292 TI - [Analysis of nine kinds of sweeteners in foods by LC/MS]. AB - A simple and rapid method for the simultaneous determination of nine kinds of sweeteners (acesulfame potassium, AK; sucralose, SUC; saccharin, SA; cyclamate, CYC; aspartame, APM; dulcin, DU; glycyrrhizic acid, GA; stevioside, STV; rebaudioside A, REB) in various foods by high-performance liquid chromatography electrospray mass spectrometry (LC/ESI-MS) was developed. The LC separation was performed on a ZORBAX Eclipse XDB-C18 (2.1 mm x 150 mm) with a mobile phase of 5 mmol/L dibutylammonium acetate (DBAA) and acetonitrile-water (8: 2). Mass spectral acquisition was done in the negative ion mode by applying selected ion monitoring (SIM). The sweeteners were extracted from foods with 0.08 mol/L phosphate buffer (pH 7.0)- ethanol (1:1), and the extract was cleaned up on a Sep pak Vac C18 cartridge after the addition of tetrabutylammonium bromide and phosphate buffer (pH 3.0). The recovery of the nine kinds of sweeteners from five kinds of foods fortified at the level 0.01 g/kg, 0.05 g/kg and 0.20 g/kg was 75.7 109.2%, and the between-day SD values were 0.5-10.9%. The quantification limits of AK, SA, CYC, APM and STV were 0.001 g/kg, and those of SUC, DU, GA and REB were 0.005 g/kg. A recovery test from each cleaned-up sample solution was necessary to detect ionization suppression. PMID- 16042293 TI - Detection of genetically modified organisms in foreign-made processed foods containing corn and potato. AB - Investigations of the validity of labeling regarding genetically modified (GM) products were conducted using polymerase chain reaction (PCR) methods for foreign made processed foods made from corn and potato purchased in the Tokyo area and in the USA. Several kinds of GM crops were detected in 12 of 32 samples of processed corn samples. More than two GM events for which safety reviews have been completed in Japan were simultaneously detected in 10 samples. GM events MON810 and Bt11 were most frequently detected in the samples by qualitative PCR methods. MON810 was detected in 11 of the 12 samples, and Bt11 was detected in 6 of the 12 samples. In addition, Roundup Ready soy was detected in one of the 12 samples. On the other hand, CBH351, for which the safety assessment was withdrawn in Japan, was not detected in any of the 12 samples. A trial quantitative analysis was performed on six of the GM maize qualitatively positive samples. The estimated amounts of GM maize in these samples ranged from 0.2 to 2.8%, except for one sample, which contained 24.1%. For this sample, the total amount found by event specific quantitative analysis was 23.8%. Additionally, Roundup Ready soy was detected in one sample of 21 potato-processed foods, although GM potatoes were not detected in any sample. PMID- 16042294 TI - [Investigation of Coxiella burnetii contamination in commercial milk and PCR method for the detection of C. burnetii in egg]. AB - A total of 244 milk samples collected from supermarkets in Tokyo were examined for contamination with Coxiella burnetii. C. burnetii DNA was detected in 131 (53.7%) of the samples by nested PCR. PCR-positive samples were injected into immunosuppressed A/J strain mice. Of the 22 PCR-positive milk samples tested, none resulted in isolation of C. burnetii from the mice. Heat-treatment was sufficient to inactivate C. burnetii in commercial milk. In addition, a PCR detection method for C. burnetii in chicken egg was developed. Egg yolk was added to an equal volume of 1 mol/L of NaCl phosphate buffer and homogenized for removal of protein and lipid. After centrifugal separation, the supernatant was removed, and template DNA in the precipitate was extracted using SDS, proteinase K and NaI. Using such prepared samples, 3.2 x 10(1) C. burnetii particles in 1 g of egg yolk could be detected by nested PCR. All of 200 chicken egg samples collected from supermarkets in Tokyo were negative for C. burnetii by the nested PCR method. PMID- 16042295 TI - [Structural analysis of unknown dyes detected in hajikami (ginger pickled in vinegar)]. AB - Hajikami (ginger pickled in vinegar) is often used as a relish for grilled fish, and it often contains coloring agents. We detected Rose Bengal (R105) and two unknown dyes in some Hajikami by thin layer chromatography. In this study, we tried to characterize these unknown dyes by HPLC with photodiode array detection (PDA-HPLC), liquid chromatography/mass spectrometry (LC/MS) and nuclear magnetic resonance (NMR). PDA-HPLC analysis showed that the spectra of the unknown dyes resembled that of R105, suggesting the molecular structures of these two dyes are similar to that of R105. Furthermore, LC/MS analysis suggested that the these dyes are R105 in which one or two iodines are replaced by hydrogen. Finally, the two dyes were determined by 1H-NMR and 13C-NMR to be 4,5,6,7-tetrachloro-3',6' dihydroxy-2',7'-diiodo spiro[isobenzofuran- 1 (3H),9'-[9H]xanthen]-3-one disodium salt and 4,5,6,7-tetrachloro-3',6'-dihydroxy-2',7'-diiodo spiro[isobenzofuran 1(3H),9'[9H]xanthen]-3-one disodium salt. PMID- 16042296 TI - [Congener-specific analysis of Pcbs in food samples by using GC/MS]. AB - A GC/MS method for congener-specific analysis of polychlorinated biphenyls (PCBs) in food samples was evaluated. There was good agreement between total concentration of PCBs in fish samples (n = 16) measured by using the GC/MS method and the conventional GC-ECD method. The total diet study samples (fishery food, meat, egg, milk, and their products) collected from Osaka in 1982-2001 were analyzed by using the GC/MS method to estimate the temporal trend of daily intake of PCBs from food. The estimated daily intake of total PCBs (sum of tri- to heptaCBs) was in the range of 0.7-4.4 microg/person/day, far below the Japanese provisional acceptable daily intake (250 microg/50 kg-person/day). The dominant congener was 2,2',4,4',5,5'-hexachlorobiphenyl (#153), which accounted for 9-15% of total PCB. The ratio of lower chlorinated congeners decreased from the 1980's to post-2000. The results indicate that the PCB congener profile in foods has gradually changed. PMID- 16042297 TI - [Metals in recycled polyethylene terephthalate and discrimination method for its use]. AB - Metals in recycled polyethylene terephthalate (PET) were analyzed by ICP-MS following microwave digestion with nitric acid. Physically and superclean-like recycled PET contained both Ge and Sb, and sometimes contained Co, P or Si. In contrast, the chemically recycled PET contained only Ge or Sb, and some samples contained Co. The recycled PETs did not contain Pb or Cd. Ge and Sb were catalysts of the polymerization, and the other metals also originated from the PET resin as additives. It was concluded that there is no safety concern about metals in recycled PET. It became clear that the presence of both Ge and Sb could identify products formed using physically or superclean-like recycled PET. According to this discrimination method, about half of the sheet molding products used recycled PET. PMID- 16042298 TI - [Identification and determination of the isomer and subsidiary color in food blue no. 2 (indigo carmine) by LC/MS and HPLC]. AB - CFR and JECFA specify that the total color in FD&C Blue No. 2 (B2; Indigo Carmine, Indigotine, Food Blue No. 2) is not less than 85%, its isomer (B2iso) in B2 is not more than 18%, and its subsidiary color (B2sub) in B2 is not more than 2% (CFR) or 1% (JECFA). Japan's Specifications and Standards for Food Additives, 7th Edition, specifies that the total color in B2 is not less than 85.0% and other color materials in B2 are not detected by paper chromatography. LC/MS and HPLC were employed to identify and determine the main component (B2m) of B2, B2iso, and B2sub. The pseudo molecular ions (B2m and B2iso: [M-2Na+H]-, m/z=421; B2sub: [M-Na]-, m/z=341) of each color material were obtained and identified by LC/MS based on their absorptions and mass spectra. The contents of B2iso and B2sub in B2 samples (certified samples from fiscal year 1998 to fiscal year 2002) were determined by HPLC using calibration curves for the standards of B2m and B2iso. The contents of B2iso in most samples were less than 10%, and the contents of B2sub in all samples were not more than 1%. All of them were within the regulatory limits set by the CFR and JECFA. PMID- 16042299 TI - Malonaldehyde content in some breads and its change during storage. AB - Malonaldehyde (MA) content in plain bread, French bread, and croissants was analyzed. MA was detected in all the analyzed samples. The MA and lipid contents in croissants were higher than those in other breads. Changes in MA content in plain bread, French bread, and croissants stored at 4 degrees C were examined after 0, 2, and 4 days of storage. The MA content in all the bread samples increased during the storage period. The highest increase of MA content was observed in French bread. PMID- 16042300 TI - Modified determination method of total bromine in agricultural products by gas chromatography. AB - In the official method for determination of total bromine in fruit and grain foods, bromine is derivatized with 3-pentanone for GC analysis. Co-existing substances sometimes interfere with measurement of the derivative, though the method is highly selective. In this study, the notification method was modified to reduce impurity peaks by applying 3-hexanone. Samples were alkalized and reduced to ash in an electric furnace. After ashing, samples were oxidized with potassium permanganate solution and derivatized with 3-hexanone. The calibration curve was linear from 0.1 microg/mL up to 5.0 microg/mL. The detection limit (S/N = 10) was 0.1 microg/mL, i.e., 5 microg/g for herb, 2.5 microg/g for grains and 1.0 microg/g for fruits. The recoveries of bromine from fruit, grain foods and herbs added at the levels of 5 to 25 microg/g ranged from 84.2 to 96.9%. The values of relative standard deviation (RSD) were from 1.4 to 6.3%. This method should be useful for routine examination of total bromine in foods. PMID- 16042301 TI - [Survey of histamine content in seafood on the market]. AB - Food poisoning involving histamine has occurred almost every year for 20 years in Tokyo, and is usually due to ingestion of fish with lean meat, such as sardine, mackerel, horse mackerel and so on. Therefore, we were investigated the levels of histamine and 4 non-volatile amines (tyramine, putrescine, cadaverine, spermidine) in 637 samples on the market. The water activity of samples in which histamine was detected at 5 mg/100 g and over was examined. Histamine, tyramine, putrescine, cadaverine and spermidine were detected in 66, 43, 26, 64 and 5 samples, and the detection ranges were 5-340, 5-51, 5-42, 5-180 and 5-8 mg/100 g, respectively. Most of the samples in which histamine was detected were semi-dried round and split sardine. Water activity of 24 samples of semi-dried round and split sardine in which histamine was detected was in the range of 0.68-0.96. PMID- 16042302 TI - [Standardization of the detection methods for genetically modified organisms in ISO]. PMID- 16042303 TI - [Guideline for the establishment of use-by date and best-before date in foods]. PMID- 16042304 TI - [Consideration of detection method and identification for genetically modified foods]. PMID- 16042305 TI - [Food poisoning caused by Zeus siquijorensis frequently occurring in China]. PMID- 16042306 TI - [Control of food-borne botulism in thermally processed low-acid foods packaged in hermetically sealed containers]. PMID- 16042307 TI - X-linked myotubular and centronuclear myopathies. AB - Recent work has significantly enhanced our understanding of the centronuclear myopathies and, in particular, myotubular myopathy. These myopathies share similar morphologic appearances with other diseases, namely the presence of hypotrophic myofibers with prominent internalized or centrally placed nuclei. Early workers suggested that this alteration represented an arrest in myofiber maturation, while other hypotheses implicated either failure in myofiber maturation or neurogenic causes. Despite similarities in morphology, distinct patterns of inheritance and some differences in clinical features have been recognized among cases. A severe form, known as X-linked myotubular myopathy (XLMTM), presents at or near birth. Affected males have profound global hypotonia and weakness, accompanied by respiratory difficulties that often require ventilation. Most of these patients die in infancy or early childhood, but some survive into later childhood or even adulthood. The responsible gene (MTM1) has been cloned; it encodes a phosphoinositide lipid phosphatase known as myotubularin that appears to be important in muscle maintenance. In autosomal recessive centronuclear myopathy (AR CNM), the onset of weakness typically occurs in infancy or early childhood. Some investigators have divided AR CNM into 3 subgroups: 1) an early-onset form with ophthalmoparesis, 2) an early-onset form without ophthalmoparesis, and 3) a late-onset form without ophthalmoparesis. Clinically, autosomal dominant CNM (AD CNM) is relatively mild and usually presents in adults with a diffuse weakness that is slowly progressive and may be accompanied by muscle hypertrophy. Overall, the autosomal disorders are not as clinically uniform as XLMTM, which has made their genetic characterization more difficult. Currently the responsible gene(s) remain unknown. This review will explore the historical evolution in understanding of these myopathies and give an update on their histopathologic features, genetics and pathogenesis. PMID- 16042308 TI - Hematopoietic cell transplantation ameliorates clinical phenotype and progression of the CNS pathology in the mouse model of late onset Krabbe disease. AB - Krabbe disease is a genetic demyelinating disease caused by a deficiency of galactosylceramidase. The majority of cases are of infantile onset with rapid clinical course. A rare late onset form with milder clinical symptoms also exists. The latter form has been reported to respond well to the bone marrow transplantation (BMT) therapy. We tested whether the BMT could be an effective therapy for the mouse model of the late onset form, saposin-A-/- (SAP-A-/-) mice. We used green fluorescent protein transgenic mice as the donors. Chimeric SAP-A-/ mice that received BMT showed very little evidence of neurologic symptoms. At postnatal day 190 when severe demyelination was evident in naive SAP-A-/- mice, demyelination was virtually absent in the brain of chimeric SAP-A-/- mice. Presence of residual enzyme activity, at the time of rapid myelination in SAP-A-/ mice, appears to limit initial inflammatory responses and macrophage infiltration, thereby preventing progression of demyelination in the CNS in SAP-A /- mice. In contrast, the peripheral nerves showed features of hypertrophic neuropathy with hypomyelination and onion bulb formation, suggesting that there are different cellular responses to the BMT in the CNS and PNS. PMID- 16042309 TI - Reinnervation of muscular targets by nerve regeneration through guidance conduits. AB - We established histopathologic and neurophysiologic approaches to examine whether different designs of polycaprolactone-engineered nerve conduits (hollow vs. laminated) could promote nerve regeneration as autologous grafts after transection of sciatic nerves. The assessments included morphometric analysis at the level of sciatic nerve, neuromuscular junction (NMJ) and gastrocnemius muscle, and nerve conduction studies on sciatic nerves. Six months after nerve grafting, the nerve fiber density in the hollow-conduit group was similar to that in the autologous-graft group; the laminated-conduit group only achieved approximately 20% of these values. The consequences of these differences were reflected in nerve growth into muscular targets; this was demonstrated by combined cholinesterase histochemistry for NMJ and immunohistochemistry for nerve fibers innervating NMJ with an axonal marker, protein gene product 9.5. Hollow conduits had similar index of NMJ innervation as autologous grafts; the values were higher than those of laminated conduits. Among the 3 groups there were same patterns of differences in the cross-sectional area of muscle fibers and amplitudes of compound muscle action potential. These results indicate that hollow conduits were as efficient as autologous grafts to facilitate nerve regeneration, and provide a multidisciplinary approach to quantitatively evaluate muscular reinnervation after nerve injury. PMID- 16042310 TI - Progression of cerebral amyloid angiopathy in transgenic mouse models of Alzheimer disease. AB - Cerebral amyloid angiopathy (CAA), the deposition of beta-amyloid (Abeta3) in cerebral vessels, has been implicated as a common cause of hemorrhagic stroke and other forms of vascular disease. CAA is also a frequent concomitant of Alzheimer disease (AD). While the longterm consequences of CAA are well recognized from clinical and pathologic studies, numerous questions remain unanswered regarding the progression of the disease. Examination of CAA in traditional histologic sections does not easily allow for characterization of CAA, particularly in leptomeningeal vessels. In order to approach this topic, we used low magnification imaging of intact, postmortem brains from transgenic mouse models of AD-like pathology to define the spatial and temporal characteristics of CAA in leptomeningeal vessels. Imaging of brains from 10- to 26-month-old animals demonstrated a stereotypical pattern to the development of CAA, with vessels over the dorsal surface of the brain showing an anterior-to-posterior and large-to small vessel gradient of involvement. High magnification imaging revealed that CAA deposition began with a banding pattern determined by the organization of the vascular smooth muscle cells. Further analysis of the pattern of amyloid deposits showed shrinkage and disappearance of the gaps between clusters of amyloid bands, gradually reaching a confluent pattern. These data led to a classification system to describe the severity of CAA deposition and demonstrate the potential of using intact brains to generate maps defining the progression and kinetics of CAA. This approach should lead to more informed analysis of the consequences of evolving therapeutic options for AD on this related form of vascular pathology. PMID- 16042311 TI - Ependymal denudation and alterations of the subventricular zone occur in human fetuses with a moderate communicating hydrocephalus. AB - In mutant rodents, ependymal denudation occurs early in fetal life, preceding the onset of a communicating hydrocephalus, and is a key event in the etiology of this disease. The present investigation was designed to obtain evidence whether or not ependymal denudation occurs in 16- to 40-week-old human fetuses developing a communicating hydrocephalus (n = 8) as compared to fetuses of similar ages with no neuropathologic alterations (n = 15). Sections through the walls of the cerebral aqueduct and lateral ventricles were processed for lectin binding and immunocytochemistry using antibodies against ependyma, astroglia, neuroblasts, and macrophages markers. Anticaveolin was used as a functional marker of the fetal ependyma. The structural and functional molecular markers are differentially expressed throughout the differentiation of the human fetal ependyma. Denudation of the ependyma of the aqueduct and lateral ventricles occurred in all fetuses developing a communicating hydrocephalus, including the youngest ones studied. The denuded surface area increased in parallel with the fetus age. The possibility is advanced that in many or most cases of human fetal hydrocephalus there is a common defect at the ependymal cell lineage leading to ependymal detachment. Evidence was obtained that in hydrocephalic human fetuses a process to repair the denuded areas takes place during the fetal life. In hydrocephalic fetuses, detachment of the ependyma of the lateral ventricles resulted in the (i) loss of the germinal ependymal zone, (ii) disorganization of the subventricular zone and, (iii) abnormal migration of neuroblasts into the ventricular cavity. Thus, detachment of the ependymal layer in hydrocephalic fetuses would not only be associated with the pathogenesis of hydrocephalus but also to abnormal neurogenesis. PMID- 16042312 TI - GluR2 deficiency accelerates motor neuron degeneration in a mouse model of amyotrophic lateral sclerosis. AB - AMPA receptor-mediated excitotoxicity has been implicated in the selective degeneration of motor neurons in amyotrophic lateral sclerosis (ALS). Motor neurons in vitro are particularly vulnerable to excessive AMPA receptor stimulation and one of the factors underlying this selective vulnerability is the presence of a large proportion of Ca2+ -permeable (i.e. GluR2-lacking) AMPA receptors. However, the precise role of GluR2-lacking AMPA receptors in motor neuron degeneration remains to be defined. We therefore studied the impact of GluR2 deficiency on motor neuron death in vitro and in vivo. Cultured motor neurons from GluR2-deficient embryos displayed an increased Ca2+ influx through AMPA receptors and an increased vulnerability to AMPA receptor-mediated excitotoxicity. We deleted the GluR2 gene in mutant SOD1G93A mice by crossbreeding them with GluR2 knockout mice. GluR2 deficiency clearly accelerated the motor neuron degeneration and shortened the life span of mutant SOD1G93A mice. These findings indicate that GluR2 plays a pivotal role in the vulnerability of motor neurons in vitro and in vivo, and that therapies that limit Ca2+ entry through AMPA receptors might be beneficial in ALS patients. PMID- 16042313 TI - Axon and Schwann cell partnership during nerve regrowth. AB - Regeneration of peripheral nerve involves an essential contribution by Schwann cells (SCs) in collaboration with regrowing axons. We examined such collaboration between new axons and Schwann cells destined to reform peripheral nerve trucks in a regeneration chamber bridging transected rat sciatic nerves. There was a highly intimate "dance" between axons that followed outgrowing and proliferating SCs. Axons without SCs only grew short distances and almost all axon processes had associated SC processes. When regeneration chambers were infused through an external access port with local mitomycin, a mitosis inhibitor, SC proliferation, migration and subsequent axon regrowth were dramatically reduced. Adding laminin to mitomycin did not reverse this regenerative lag and indicated that SCs provide more than laminin synthesis alone. Laminin infused alone supplemented endogenous laminin and facilitated first SC then axon regrowth. "Wrong way" misdirected axons were associated with misdirected SC processes and were more numerous in bridges exposed to mitomycin, but were fewer in laminin supplemented bridges. Later, by 21 days, there was myelinated axon repopulation of regenerative bridges but those exposed to mitomycin alone at early time points had substantial impairments in axon investment. Reforming peripheral nerve trucks involves a very close and intimate relationship between axons and SCs that must proliferate and migrate, facilitated by laminin. PMID- 16042314 TI - Staging of sporadic Parkinson disease-related alpha-synuclein pathology: inter- and intra-rater reliability. AB - Sporadic Parkinson disease (sPD) is characterized by alpha-synuclein (alpha-syn) inclusions. The distribution of such inclusions appears to relate to disease progression and severity. We propose and test a simple staging protocol based on the presence of alpha-syn immunoreactivity in 5 paraffin sections that, taken together, contain up to 8 vulnerable brain regions. Six stages of alpha-syn pathology reminiscent for sPD are defined based on the presence or absence of inclusions in the assessed sections. Six observers from 5 different institutions rated 21 cases on the basis of written instructions only. The agreement of the raters was highly significant with a mean error below one stage. Both inter- and intra-rater reliability were also substantial to almost perfect as analyzed by paired comparison between all raters. We propose that the staging procedure for alpha-syn pathology is suitable for application in routine neuropathology and brain banking. Clearly defined stages of alpha-synpathology might aid the comparability between studies and also help to distinguish sPD from other synucleinopathies. PMID- 16042315 TI - Distinct allelic variants of TSC1 and TSC2 in epilepsy-associated cortical malformations without balloon cells. AB - Epilepsy-associated malformations of cortical development (MCDs) comprise a variety of dysplastic and neoplastic lesions of yet undetermined molecular pathology. Histopathologic similarities between MCDs and dysplastic brain lesions in the autosomal inherited neurocutaneous phacomatosis tuberous sclerosis (TSC), which affects the TSC1 and/or TSC2 genes, suggest common pathogenetic mechanisms. Previous studies revealed different alterations of TSC1 and TSC2 in epilepsy associated malformations and glio-neuronal tumors despite histopathologic similarities. In order to examine current clinico-pathologic classification systems of cortical malformations on the molecular level, we carried out a mutational analysis of TSC1 and TSC2 in a series of surgical specimens obtained from patients with FCD without Taylor type balloon cells (FCDIIa; n = 20), architectural dysplasias (FCDI; n = 15), nodular cortical heterotopias (NCH; n = 4), and heterotopic white matter neurons (WMNH; n = 19). In FCDIIa, abundant genomic polymorphisms were detected in TSC2 (intron 4) but no allelic variants observed in exon 17 of TSC1. This allelic distribution pattern is in contrast to findings in FCDI and WMNH but also to those previously reported in FCDIIb (Taylor's balloon cell type). The latter revealed increased frequencies of specific alleles only in TSC1. The determination of characteristic molecular genetic alterations in specific epilepsy-associated malformations will support a comprehensive clinico-pathologic classification system and help to identify molecular pathways with potential pathogenetic relevance. Our work is supported by DFG (SFB TR3 [AJB], DFG Bl 421/1-1 [IB]), BONFOR, and Deutsche Krebshilfe. PMID- 16042316 TI - Abnormal group I metabotropic glutamate receptor expression and signaling in the frontal cortex in Pick disease. AB - Group I metabotropic glutamate receptors (mGluR1) regulate synaptic transmission through the stimulation of phospholipase Cbeta1 (PLCbeta1) and then by the activation of protein kinase C (PKC). Considering these properties, it is conceivable that major cortical functional deficits may be attributed to abnormal mGluR processing and signaling. The present work examines mGluRI expression and signaling in the frontal cortex (area 8) of 3 cases with Pick disease (PiD), a neurodegenerative disease with abnormal phospho-tau accumulation, in comparison with 3 age-matched controls by means of glutamate binding assays, enzymatic activity, gel electrophoresis and Western blotting, solubility and immunoprecipitation assays, and confocal microscopy. Reduced expression levels of PLCbeta1 and reduced PLCbeta1 activity have been found in PiD. The expression levels of the nonrelated phospholipase PLCgamma, a substrate of tyrosine kinase, are also reduced in PiD. This is accompanied by a marked decrease in the expression of cPKCalpha and increased expression of the inner band (76 kDa) of the nPKCdelta doublet at the expense of a decrease of the phosphorylated (active) form (78 kDa). In contrast, L-[3H]glutamate-specific binding to mGluRs is augmented in PiD cases, mainly because of the higher mGluR1 and mGluRs expression levels detected. No modifications in PLCbeta1 solubility have been observed in PiD and no interactions between PLCbeta1 and tau have been demonstrated in diseased and control cases. Moreover, double-labeling immunofluorescence and confocal microscopy have shown no colocalization of phospho-tau (AT8 antibody) and PLCbeta1 in phospho-tau inclusions, including Pick bodies. These results demontrate for the first time abnormal mGluR signaling in the cerebral cortex in PiD and selective vulnerability of phospholipases and PKC to PiD. PMID- 16042317 TI - Sporadic phaeochromocytoma in childhood: clinical and molecular variability. AB - Sporadic phaeochromocytoma is an infrequent tumour during paediatric age and may or may not be associated with specific autosomal dominant inherited cancer syndromes such as multiple endocrine neoplasia type 2 (MEN2), von Hippel-Lindau syndrome (VHL) type 2 or neurofibromatosis (NF) type 1. We report two cases of benign, adrenal, and unilateral phaeochromocytoma that clearly demonstrate the clinical and molecular heterogeneity of this disease during the paediatric period. The first patient presented a characteristic symptomatic form of sporadic phaeochromocytoma. The second patient, an incidental finding, was practically asymptomatic and had a de novo germline point mutation in the VHL gene (Arg167Trp). The frequency of de novo mutations in susceptible genes (especially the VHL gene) in paediatric patients with sporadic phaeochromocytoma and the elevated mortality of these cancer syndromes suggest that screening for mutations should be performed even in cases of non-familial sporadic phaeochromocytoma. PMID- 16042318 TI - Postponing puberty in children with growth hormone deficiency. PMID- 16042319 TI - Gonadotropin releasing hormone analogue therapy in children with isolated growth hormone deficiency: final height benefit from postponing puberty? AB - Although growth hormone (GH) treatment has improved final height prognosis in children with GH deficiency (GHD), adult heights are still disappointing. Final height could be improved by increasing the duration of puberty and in this way increasing total pubertal height gain. Many studies have been published on the effect of gonadal suppression, mostly by gonadotropin releasing hormone (GnRH) analogues, on final height in children with GHD. Because of the different methodologies used in these studies, results are difficult to compare. Both positive and marginal effects on final height have been reported; however, patient numbers are limited. Children with GHD who start puberty at a relatively young age and who have a poor predicted adult height, can benefit from the addition of GnRH analogues. From previous studies, we might conclude that when there is a positive effect, height benefit is marginal. However, additional prospective, randomized controlled trials are needed to further elucidate whether delaying puberty is indicated in children with GHD to improve final height. PMID- 16042320 TI - Kallmann's syndrome: bridging the gaps. PMID- 16042321 TI - Kallmann's syndrome with a novel missense mutation in the KAL1 gene that modifies the major cell adhesion site of the anosmin-1 protein. AB - Kallmann's syndrome (KS) refers to the association of hypogonadic hypogonadism and anosmia or hyposmia. The X-linked form of the disease is due to mutations in the KAL1 gene that encodes for the protein anosmin-1. We studied the KAL1 gene in a patient with KS and his family by PCR amplification and direct sequencing. A novel missense mutation (V263G) that modifies the major cell adhesion site of the anosmin-1 protein was identified. Our results suggest that this reported mutation is responsible for KS and might help to elucidate the function of an important area of the anosmin-1 protein. PMID- 16042322 TI - Gender identity and sex-of-rearing in children with disorders of sexual differentiation. AB - AIM: To compare declared sexual identity to sex-of-rearing in individuals with disorders of sexual differentiation. METHODS: All 84 patients > or =5 years old in a pediatric psychosexual development clinic were assessed for sex-of-rearing and sexual identity. Diagnoses included 1) male-typical prenatal androgen effects but an absent or severely inadequate penis - 45 patients with cloacal exstrophy or aphallia; 2) inadequate prenatal androgens and a Y-chromosome - 28 patients with partial androgen insensitivity (pAIS), mixed gonadal dysgenesis (MGD), hermaphroditism, or craniofacial anomalies with genital ambiguity; 3) inappropriate prenatal androgen effects and a 46,XX karyotype - 11 patients with congenital adrenal hyperplasia (CAH). RESULTS: Of 73 patients with disordered sexual differentiation and a Y-chromosome, 60 were reared female; 26 of the 60 (43%) declared female identity while 32 (53%) declared male identity including 18 (55%) with cloacal exstrophy, six (55%) with MGD, four (40%) with pAIS, one (50%) with aphallia, one (100%) with hermaphroditism, and two (67%) with craniofacial anomalies; two (3%) declined to discuss identity. Nine of 11 patients with CAH and a 46,XX karyotype were reared female and two reared male; six (55%) declared female identity and five (45%) declared male identity. Of 84 total patients, 69 were reared female, but only 32 lived as female, while 29 lived as male; four patients refused to discuss sex-of-living; parents of four patients rejected their declarations of male identity. All 15 patients reared male lived as male including two genetic females. CONCLUSION: Active prenatal androgen effects appeared to dramatically increase the likelihood of recognition of male sexual identity independent of sex-of-rearing. Genetic males with male-typical prenatal androgen effects should be reared male. PMID- 16042323 TI - A long-term outcome study of intersex conditions. AB - CONTEXT: Clinical management of intersex conditions is controversial because the available evidence is limited and conflicting, with no long-term population based studies comparing matched controls. OBJECTIVE: To assess the long-term psychological, sexual and social outcomes of patients with intersex compared with two matched control populations. DESIGN, SETTING AND PARTICIPANTS: Three different aged-matched (18-32 years) patient groups completed a self-administered questionnaire of established quality of life and well-being inventories measuring physical' health, psychological adjustment and sexuality, following a mail-out to all identified patients. The intersex group (n = 50) and the Hirschsprung disease, a congenital disorder, control group (n = 27), were patients who had attended the Royal Children's Hospital, a tertiary centre, for their clinical care. The insulin dependent diabetes mellitus control group was recruited from an adult tertiary hospital. The study was conducted at the hospital-based Murdoch Childrens Research Institute. MAIN OUTCOME MEASURES: Psychological, sexual and social outcomes. RESULTS: The intersex group did not differ from controls on physical or mental health, depression, state anxiety, neuroticism, psychoticism or stressful life events. Intersex participants were satisfied with their overall body appearance, although intersex males were less satisfied than controls with the size (p <0.05) and appearance (p <0.01) of their sex organs. The intersex group was less likely to experience orgasm (p <0.05), tended to experience more pain during intercourse (p = 0.06), had more difficulties with penetration (p <0.01) and were less likely to have sexual activity several times or more a week (p <0.05) than the combined control groups. Intersex participants did not differ from controls in level of sexual desire or enjoyment of sexual activities. CONCLUSIONS: Most patients with intersex had positive psychosocial and psychosexual outcomes, although some problems were reported with sexual activity. These results overall suggest that a model of care including early genital surgery carried out at a centre of excellence with a multidisciplinary team can minimize long-term complication rates. PMID- 16042324 TI - Idiopathic male pseudohermaphroditism: variations in presentation and management. AB - Male pseudohermaphroditism (MPH) is the abnormal development of genitalia in an individual with a 46,XY chromosome complement and testicular tissue. The etiology of MPH is unknown in most cases, which are defined as idiopathic. OBJECTIVE: To analyze the data for cases of idiopathic MPH. PATIENTS AND METHODS: A retrospective study of 29 patients with idiopathic MPH and no uterus. RESULTS: Four patients had a family history of abnormal sexual development and five had low birth weight. The initial manifestations were sexual ambiguity (26), microphallus and hypospadias (2), and primary amenorrhea (1). Basal and/or stimulated testosterone concentrations showed insufficient testosterone secretion in three patients. Genitography showed a vagina in 13 patients. Male genitoplasties were performed on 21 out of the 24 patients reared as males and female genitoplasties on five patients. Histological studies of the gonads of these showed streak gonads in one, normal gonads in one and signs of testicular dysgenesis in three others. Molecular studies on the SRY gene (17) showed no mutation. CONCLUSIONS: Idiopathic male pseudohermaphroditism is a heterogeneous condition, even within families with a history of this condition. We propose a set of guidelines for the management of these patients. PMID- 16042325 TI - Assessment of pubertal development in Egyptian girls. AB - Puberty is a significant event of human growth and maturation associated with marked physiological and psychological changes. The aim of this study was to assess normal pubertal development in Egyptian girls to define normal, precocious and delayed puberty. The present study included a cross-sectional sample of 1,550 normal Egyptian girls of high and middle socioeconomic class living in Cairo. Their ages ranged from 6.5 to 18.5 years. Pubertal assessment was made according to Tanner staging. The mean menarcheal age (MMA) was estimated using probit analysis. Weight and height were measured and body mass index (BMI) was calculated. The mean age at breast bud stage (B2) was 10.71+/-1.6, pubic hair stage (PH2) was 10.46+/-1.36, while axillary hair stage (A2) was 11.65+/-1.62 and MMA was 12.44 years. The mean age at attainment of puberty was compared with those of other Egyptian studies and other populations. Girls of the present study started pubertal development and achieved menarche earlier than those of previous Egyptian studies confirming a secular trend. Differences between the present study and other worldwide studies can be attributed to various genetic, racial, geographical, nutritional, and secular trend factors. PMID- 16042326 TI - Antiepileptic drug-induced osteopenia in ambulatory epileptic children receiving a standard vitamin D3 supplement. AB - Dual energy X-ray absorptiometry (DEXA) is a non-invasive, rapid, accurate and highly reproducible method for the assessment of antiepileptic drug (AED)-induced osteopenia in epileptic children. In this study, we investigated bone mineral density (BMD) using DEXA in 56 epileptic children receiving long-term AED treatment for at least 2 years. All children received AED monotherapy or polytherapy plus a standard vitamin D3 supplement (400 U/day). BMD measurements were made from lumbar spine (L2-L4) regions. Age- and sex-specific BMD SD scores were calculated for each child. Osteopenia was defined as SD scores less than 1.5. There was no significant difference in mean BMD values between epileptic children receiving monotherapy or polytherapy. The results were also compared to the age- and sex-specific BMD SD scores obtained from healthy Turkish children. Only three patients (5%) receiving AED therapy had a BMD SD score less than -1.5. This rate is relatively lower than the rates of previous studies conducted on ambulatory children on long-term AED treatment without vitamin D3 supplementation. PMID- 16042327 TI - Growth hormone improves bone mineral content in children with cystic fibrosis. AB - AIM: Osteoporosis and osteopenia have been reported as common complications of cystic fibrosis (CF); however, little is known about accrual of bone mineral in CF. The goal of our study was to measure bone mineral content (BMC) in non acutely-ill, but poorly growing children with CF, and to determine the relationship between height, lean body mass and BMC. Our second aim was to evaluate the effect of one year of treatment with human recombinant growth hormone (GH) on total body BMC. METHODS: We measured total-body BMC using dual energy X-ray absorptiometry in 32 poorly growing (height < or =10th percentile for age) prepubertal Caucasian children (ages 7 years 6 months-12 years 9 months, 17 M and 15, F) with CF. BMC and lean tissue mass (LTM) were measured at baseline, at 6 months and one year. One half of the children were randomly assigned to receive treatment with GH (GHTX). Results were compared to reference data maintained for healthy children matched for age and ethnicity. Sex steroid and IGF-I levels were also measured. RESULTS: Children with CF exhibited lower total body BMC and LTM than age-, ethnicity- and gender-matched controls. This was still apparent when the data were matched for height and bone age. BMC correlated with height, LTM, and IGF-I levels. Although at baseline the groups were similar, the GHTX group demonstrated significantly greater increase in height, weight, LTM and BMC than the NonTX group. These differences remained despite correction for increase in height CONCLUSION: Our study is the first to evaluate BMC in children with CF and suggests that poor accumulation of bone mineral is a problem. We have further demonstrated that GH treatment improves accumulation of bone mineral. PMID- 16042328 TI - Effective methimazole dose for childhood Graves' disease and use of free triiodothyronine combined with concurrent thyroid-stimulating hormone level to identify mild hyperthyroidism and delayed pituitary recovery. AB - Appropriate methimazole dosing for initial treatment of childhood Graves' disease is uncertain. A retrospective chart review was performed on 5 to 17 year-old children treated for Graves' disease. Patients were divided into two groups depending on initial methimazole dosing: low-dose and high-dose regimens using <0.5 mg/kg/day and >0.5 mg/kg/day, respectively. The low-dose regimen was effective in 5/12 (42%) of patients and the high-dose regimen was effective in 27/33 (82%) of patients (p = 0.016). There was also a statistically significant dose/time interaction for levels of free thyroxine (T4) (p = 0.025). During treatment, 63.3% of diagnosable samples showed unambiguous hyperthyroidism or triiodothyronine (T3) toxicosis, 16.7% elevated free T3 with normal free T4 and T3 levels, indicating borderline hyperthyroidism, and 20% showed thyroid stimulating hormone (TSH) suppression with normal or low levels of free T4 and free T3, indicating delayed recovery of pituitary TSH secretion. Free T3 levels combined with concurrent TSH levels permit differentiation of mild hyperthyroidism from delayed pituitary recovery. PMID- 16042329 TI - True hermaphroditism with characteristics of Klinefelter's syndrome: a rare presentation. AB - True hermaphroditism, a very rare cause of intersex, is usually diagnosed during the newborn period in the course of evaluating ambiguous genitalia. As an exception we report an unusual case of a 14.5 year-old boy with phenotypically near-normal male genitalia and bilaterally descended gonads, who was seen for evaluation of gynecomastia and hematuria. His eunuchoid body habitus and mild mental retardation were compatible with Klinefelter's syndrome. He had a low level of free testosterone (15.2 pmol/l), and high level of estradiol (264.3 pmol/l) for his age. The patient was diagnosed as true hermaphroditism with 46,XX /47,XXY karyotype causing an ovotestis with inguinal uterus hernia in the left scrotum and a dysgenetic testis in the right scrotum. PMID- 16042330 TI - Isolated adrenocorticotropin deficiency in a child with Kabuki syndrome. AB - A 6 year-old Chinese boy with Kabuki syndrome presented with hypoglycemic seizure. He was diagnosed to have isolated adrenocorticotropin deficiency. To our knowledge, this is the first case of Kabuki syndrome with isolated adrenocorticotropin deficiency in the literature. PMID- 16042331 TI - Hemorrhagic pituitary apoplexy in an 18 year-old male presenting as non-ketotic hyperglycemic coma (NKHC). AB - Pituitary apoplexy is an acute clinical event usually caused by hemorrhage or infarction in a pituitary adenoma. We report the unusual case of hemorrhagic pituitary apoplexy in an 18 year-old male with previously undiagnosed type 2 diabetes mellitus who presented with unexplained hyperglycemia (glucose 49.2 mmol/l [887 mg/dl]) and obtundation and in whom an initial diagnosis of non ketotic hyperglycemic coma (NKHC) was made. MRI revealed a heterogeneous mass arising from an expanded sella turcica into the suprasellar cistern. Despite well controlled glucose levels on continuous insulin infusion, dexamethasone, and initiation of bromoergocriptine (parlodel) therapy, the patient's vision and pupillary responses deteriorated acutely. Following emergency transphenoidal surgery, the patient's vision and mental status improved. Data confirmed preoperative panhypopituitarism; serum prolactin was 396 ng/ml (microg/l). Immunostudies demonstrated tumoral labeling for prolactin, but not for ACTH, GH, TSH, LH, FSH, or P53. PMID- 16042332 TI - Secondary diabetes mellitus: late complication of glycogen storage disease type 1b. AB - We report a 22 year-old male with glycogen storage disease type 1 (GSD-1) who developed diabetes mellitus secondary to pancreatic islet beta-cell insufficiency. Diabetes mellitus should be considered among the late complications of GSD-1. The pathogenesis of the conversion, from a disease characterized by hypoglycemia to a disease dominated by hyperglycemia, is discussed. PMID- 16042333 TI - Neonatal hypothyroidism due to maternal vegan diet. PMID- 16042334 TI - Chemical composition of hips essential oils of some Rosa L. species. AB - The chemical composition of the hips essential oils of 9 taxa of Rosa L. was analyzed and compared using the standardized analytical GC and GC/MS methods. The volatile hips oil compositions for these species are presented. All oil samples were dominated by following components: vitispiran (isomer), a-E-acaridial, dodecanoic acid, hexadecanoic acid, docosane (C22), beta-ionone, 6-methyl-5 hepten-2-one, 2-heptanone, heptanal, myristic acid and linolic acid. Statistical analyses of 97 GC peaks of these oils were used to distinguish compositional patterns. There appeared to be correlation between the essential oil patterns and the classification within Rosa L. Cluster analysis of the composition of main components clearly showed two groups, one constituted by R. rugosa Thunb. from the Cinnamomea section, and the other constituted by the remaining taxa from the Caninae section. PMID- 16042335 TI - Flavonol glycosides from distilled petals of Rosa damascena Mill. AB - Flavonol glycosides were extracted from petals of Rosa damascena Mill. after industrial distillation for essential oil recovery and characterized by high performance liquid chromatography-electrospray ionization mass spectrometry. Among the 22 major compounds analyzed, only kaempferol and quercetin glycosides were detected. To the best of our knowledge, the presence of quercetin 3-O galactoside and quercetin 3-O-xyloside has so far not been reported within the genus Rosa. In addition, based on their fragmentation patterns, several acylated quercetin and kaempferol glycosides, some of them being disaccharides, were identified for the first time. The kaempferol glycosides, along with the kaempferol aglycone, accounted for 80% of the total compounds that were quantified, with kaempferol 3-O-glucoside being the predominant component. The high flavonol content of approximately 16 g/kg on a dry weight basis revealed that distilled rose petals represent a promising source of phenolic compounds which might be used as functional food ingredients, as natural antioxidants or as color enhancers. PMID- 16042336 TI - Antimicrobial activity of isopteropodine. AB - Bioassay-directed fractionation for the determination of antimicrobial activity of Uncaria tomentosa, has led to the isolation of isopteropodine (0.3%), a known Uncaria pentacyclic oxindol alkaloid that exhibited antibacterial activity against Gram positive bacteria. PMID- 16042337 TI - 1,4-benzoxazin-3-one, 2-benzoxazolinone and gallic acid from Calceolaria thyrsiflora Graham and their antibacterial activity. AB - Secondary metabolites, DIBOA, HBOA, 7-OH-HBOA, BOA and gallic acid, were isolated and quantified from Calceolaria thyrsiflora Graham, a native medicinal plant of Chile belonging to the Scrophulariaceae family. The highest DIBOA contents were determined in leaves (145 mmol kg(-1) dry wt) and flowers (161 mmol kg(-1) dry wt). Antibacterial activities of DIBOA, HBOA, BOA, gallic acid and infusions of flowers and leaves were determined. The phytomedicinal properties attributed to C. thyrsiflora Graham could be understood on the basis of its antibacterial activity. PMID- 16042338 TI - Bioactivity of biflorin, a typical o-naphthoquinone isolated from Capraria biflora L. AB - Capraria biflora L. (Scrophulariaceae) is a perennial shrub widely distributed in several countries of tropical America. The present work verified the cytotoxic and antioxidant potential of biflorin, an o-naphthoquinone isolated from C. biflora collected in the northeast region of Brazil. The cytotoxicity was tested on three different animal cell models: mouse erythrocytes, sea urchin embryos and tumor cells, while the antioxidant activity was assayed by the thiocyanate method. Biflorin lacked activity on mouse erythrocytes as well as on the development of sea urchin eggs, but strongly inhibited the growth of all five tested tumor cell lines, especially the skin, breast and colon cancer cells with IC50 of 0.40, 0.43 and 0.88 micro/ml for B16, MCF-7 and HCT-8, respectively. Biflorin also showed potent antioxidant activity against autoxidation of oleic acid in a water/alcohol system. PMID- 16042339 TI - Antioxidant activity of anthocyanin glycoside derivatives evaluated by the inhibition of liposome oxidation. AB - Cyanidin-3-glycosides (arabinoside, rutinoside, galactoside and glucoside) and delphinidin-3-rutinoside were examined for their ability to inhibit lipid peroxidation induced either by Fe(II) ions, UV irradiation or 2,2'-azobis(2 amidinopropane) dihydrochloride (AAPH) peroxyl radicals in a liposomal membrane system. The antioxidant abilities of anthocyanins were compared with a water soluble tocopherol derivative, trolox. The antioxidant efficacies of these compounds were evaluated by their ability to inhibit the fluorescence intensity decay of the extrinsic probe 3-[p-(6-phenyl)-1,3,5,-hexatrienyl] phenylpropionic acid, caused by the free radicals generated during peroxidation. All the anthocyanins tested (at concentrations of 15-20 microM) exhibited higher antioxidant activities against Fe(II)-induced peroxidation than UV- and AAPH induced peroxidation, suggesting that metal chelation may play an important role in determining the antioxidant potency of these compounds. It was also found that delphinidin-3-rutinoside had a higher antioxidant activity against Fe(II)-induced liposome oxidation than cyanidin-3-rutinoside, which indicates an important role of the OH group in the B ring of delphinidin-3-rutinoside in its antioxidant action. The antioxidant activity of all the anthocyanins studied was higher than that of trolox in the case of Fe(II)-induced liposome oxidation and was comparable with the action of trolox in the case of UV- and AAPH-induced liposome membrane oxidation. PMID- 16042340 TI - Separation of bioactive biflavonoids from Rheedia gardneriana using chitosan modified with benzaldehyde. AB - This paper shows the influence of benzenic groups on the chitosan surface for the separation of bioactive biflavonoids from Rheedia gardneriana leaves The yield of the biflavonoids using chitin modified with benzaldehyde (CH-Bz) as adsorbent in column chromatography was higher than that achieved with silica gel and chitosan. The presence of benzenic groups decreases the polarity of chitosan and consequently the interaction of hydrogen bonding between phenolic hydroxyl (OH) of biflavonoids and amine groups of the adsorbent. Therefore, the separation of these compounds appears to be the result of hydrophobicity and pi-pi interaction among electrons from the aromatic ring in sorbent and biflavonoid molecules. PMID- 16042341 TI - Insecticidal activity of the essential oil of Ligusticum mutellina roots. AB - The essential oil obtained from roots of different collections of Ligusticum mutellina was tested against 3rd instar armyworms, Pseudaletia unipuncta (Lepidoptera: Noctuidae), for insecticidal activity. The main compounds were isolated and their structures were elucidated using 2D-NMR techniques. Our collections contained dillapiole, ligustilide and myristicin as major compounds. The previously reported sarisan was not present, moreover its occurrence in L. mutellina should be revised based on our findings. PMID- 16042342 TI - Inhibition of functionalized 1,3-dienes against Trypanosoma cruzi. AB - Six functionalized 1,3-dienes were synthesized using cross-coupling reactions, catalyzed by palladium complexes, between alkenylboronic acids and alpha-bromo alpha,beta-unsaturated carbonylic compounds. Their cytotoxicity against epimastigotes of Trypanosoma cruzi and fibroblastic Vero cells was evaluated, using concentrations ranging from 100 microM to 2.5 mM in experiments with three incubation times (4, 8 and 16 h). These tests were performed using MTT [3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] colorimetric bioassays and its further reduction to formazan, according to the viability of the parasites and cells. With the exception of (5E,6E)-5-benzylidene-2-methylundec-6-en-4-one, all compounds were cytotoxic to both Trypanosoma cruzi and Vero cells, however differential values of IC50 were observed for two of these compounds. A possible structure-activity relationship is discussed. PMID- 16042343 TI - Assessment of potential application of binary mixtures of 2,4-d with novel aminophosphonates. AB - A series of new aminoalkane- and aminofluorenephosphonates was synthesized for agrochemical application. The particular compounds had different alkyl substituents at the carbon, nitrogen and phosphorus atoms. Their pesticidal activity was checked by applying various experimental methods. These included the measurements of compounds' potency: to inhibit growth of cucumber and germination of white mustard seeds, to influence on the membrane potential of algae and to damage human erythrocyte membranes resulting in hemolysis. All the aminophosphonates were also used in equimolar binary mixtures with the well-known herbicide 2,4-dichlorophenoxyacetic acid (2,4-D), to check, if using such mixtures, the biological efficiencies found for particular compounds could be enhanced due to interactions between aminophosphonates and 2,4-D. The results demonstrated, that depending on the structural features of the compounds, the final effects differed from antagonistic, through additive to the most promising synergistic ones. However, the type of interaction between 2,4-D and the compounds studied found in different experiments was somewhat different. In order to estimate those effects various statistical methods were used (toxic unit method, isobole method). PMID- 16042344 TI - The possible role of hydroxylation in the detoxification of atrazine in mature vetiver (Chrysopogon zizanioides Nash) grown in hydroponics. AB - The resistance mechanism of vetiver (Chrysopogon zizanioides) to atrazine was investigated to evaluate its potential for phytoremediation of environment contaminated with the herbicide. Plants known to metabolise atrazine rely on hydroxylation mediated by benzoxazinones, conjugation catalyzed by glutathione-S transferases and dealkylation probably mediated by cytochromes P450. All three possibilities were explored in mature vetiver grown in hydroponics during this research project. Here we report on the chemical role of benzoxazinones in the transformation of atrazine. Fresh vetiver roots and leaves were cut to extract and study their content in benzoxazinones known to hydroxylate atrazine, such as 2,4-dihydroxy-2H-1,4-benzoxazin-3(4H)-one (DIBOA), 2,4-dihydroxy-7-methoxy-2H-1,4 benzoxazin-3(4H)-one (DIMBOA) and their mono- and di-glucosylated forms. Identification of benzoxazinones was performed by thin layer chromatography (TLC) and comparison of retention factors (Rf) and UV spectra with standards: although some products exhibited the same Rf as standards, UV spectra were different. Furthermore, in vitro hydroxylation of atrazine could not be detected in the presence of vetiver extracts. Finally, vetiver organs exposed to [14C]-atrazine did not produce any significant amount of hydroxylated products, such as hydroxyatrazine (HATR), hydroxy-deethylatrazine (HDEA), and hydroxy deisopropylatrazine (HDIA). Altogether, these metabolic features suggest that hydroxylation was not a major metabolic pathway of atrazine in vetiver. PMID- 16042345 TI - Photochemical and antioxidant responses in the leaves of Xerophyta viscosa Baker and Digitaria sanguinalis L. under water deficit. AB - In this study, photochemical and antioxidant responses of the monocotyledonous resurrection plant Xerophyta viscosa Baker and the crab grass Digitaria sanguinalis L. under water deficit were investigated as a function of time. Water deficit was imposed by withholding irrigation for 21 d. Gas exchange and chlorophyll a fluorescence analyses indicated that the dehydration treatment caused photoinhibition in both species. The reduction in the photosynthesis rate in both species during water deficit probably contributed to the decline in the photochemical efficiency of PSII and electron transport rate. However, the stomatal conductance of both species did not change during treatment whereas the intercellular CO2 pressure increased after 10 d of water deficit treatment. These observations could be related to nonstomatal limitations. The increasing net transpiration rate of both species may have contributed to leaf cooling because of water limitations. Prolonged water deficit resulted in photosynthetic pigment chlorophyll (a + b) and carotenoids content loss in only D. sanguinalis. Both species especially D. sanguinalis had increased the level of anthocyanin after 15 d of treatment, possibly to prevent the damaging effect of photooxidation. The total SOD activity of D. sanguinalis was significantly different from X. viscosa during the treatment. The total peroxidase activity in D. sanguinalis was significantly higher than in X. viscosa. X. viscosa acclimated to water deficit with no ultimate apparent oxidative damage due to endogenous protective mechanisms of resurrection. In case of D. sanguinalis, water deficit induced considerable stress and possibly caused some oxidative damage, despite the upregulation of protection mechanisms. PMID- 16042346 TI - Effect of salt stress on the production and properties of extracellular polysaccharides produced by Cryptococcus laurentii. AB - The composition, main structural features and molecular properties of exopolysaccharides (EP) produced by Cryptococcus laurentii var. laurentii CCY 17 3-16 under optimal (EPo) and NaCI-stress conditions (EPs) as well as their subfractions isolated by gel chromatography were studied using chemical, FT-IR and NMR spectroscopy methods. The results showed that under stress conditions the yeast produced EP with a lower content of protein and phosphorus. In comparison to EPo, the EPs exhibited a substantially larger proportion of high molecular mass populations. NMR analysis of EPs revealed a higher degree of branching with single xylose side chains of the heteromannan components. The increase of the molecular mass and degree of branching of the macromolecular chains of the heteromannan components might in part be related to the function of EPs to protect the yeast cells from water loss and maintain growth conditions under the salt stress. PMID- 16042347 TI - Beta-carboline and quinoline alkaloids in root cultures and intact plants of Peganum harmala. AB - Alkaloid profiles of root and shoot cultures, seedlings and mature plants were analysed by capillary GLC and GLC-MS. beta-Carboline alkaloids, such as harmine, harmaline dominate in normal and root cultures transformed by Agrobacterium rhizogenes, as well as in roots and fruits of the plant. In shoots, flowers and shoot cultures quinoline alkaloids such as peganine, deoxypeganine, vasicinone and deoxyvasicinone widely replace the beta-carboline alkaloids. In root cultures, the formation of beta-carboline alkaloids can be induced by methyljasmonate and several other elicitors indicating that these alkaloids are part of the reactive chemical defence system of Peganum harmala. PMID- 16042348 TI - Regio- and stereoselective fungal oxyfunctionalisation of limonenes. AB - Selective transformations of limonene by asco- and basidiomycetes were investigated. On the shake flask scale, Penicillium citrinum hydrated R-(+) limonene to a-terpineol [83% regioselectivity (rs), more than 80 mg l(-1) product yield], and Gongronella butleri catalysed the terminal oxidation to yield perillyl alcohol (60% rs, 16 mg l(-1)). On the laboratory bioreactor scale, Penicillium digitatum produced a peak concentration of 506 mg a-terpineol l(-1) in the fed-batch mode, equivalent to a theoretical yield of 67%, and no volatile by-products were found. Fusarium proliferatum transformed R-(+)-limonene enantiospecifically to cis-(+)-carveol (98.6% ee, more than 35 mg l(-1) product yield) and S-(-)-limonene predominantly to trans-(-)-carveol (96.3% ee). Pleurotus sapidus selectively dehydrogenised the accumulating trans-(-)-carveol to the corresponding enantiopure R-(-)-carvone. The results show that a careful selection of strain and bioprocess parameters may improve both the yield and the optical purity of a desired product. PMID- 16042349 TI - Metabolites from endophytes of the medicinal plant Erythrina crista-galli. AB - Erythrina crista-galli (Fabaceae) is used in Argentinean ethnopharmacology as anti-inflammatory medication, narcotic, desinfectant, and for the treatment of wounds. The common name of the tree is "ceibo" or coral tree. The dominating endophytes in E. crista-galli all belong to the genus Phomopsis as identified by microscopic features and the analysis of their ITS sequences. To investigate a possible contribution of Phomopsis spp. to the metabolites found in the plant, twelve different isolates were cultivated in different media. Besides several new metabolites a number of known compounds were detected: mellein, nectriapyrone, 4 hydroxymellein, scytalone, tyrosol, clavatol, mevinic acid, and mevalonolactone. PMID- 16042350 TI - Inhibition of TNF-alpha promoter activity and synthesis by A11-99-1, a new cyclopentenone from the ascomycete Mollisia melaleuca. AB - In a search for inhibitors of the inducible tumor necrosis factor-a (TNF-a) promoter activity and synthesis, a new chlorinated cyclopentenone was isolated from fermentations of the ascomycete Mollisia melaleuca. The structure was determined by a combination of spectroscopic techniques The compound blocked the inducible human TNF-alpha promoter activity and synthesis with IC50-values of 2.5 5 microg/ml (8.1-16.1 microM). Studies on the mode of action of the compound revealed that the inhibition of TNF-a promoter activity is caused by an inhibition of the phosphorylation of the IkappaB protein which prevents the activation of the transcription factor NF-xB. No cytotoxic, antibacterial and antifungal activities could be observed up to 100 microg/ml (323 microM) of the compound. PMID- 16042351 TI - Non-homologous DNA end joining repair in normal and leukemic cells depends on the substrate ends. AB - Double-strand breaks (DSBs) are the most serious DNA damage which, if unrepaired or misrepaired, may lead to cell death, genomic instability or cancer transformation. In human cells they can be repaired mainly by non-homologous DNA end joining (NHEJ). The efficacy of NHEJ pathway was examined in normal human lymphocytes and K562 myeloid leukemic cells expressing the BCR/ABL oncogenic tyrosine kinase activity and lacking p53 tumor suppressor protein. In our studies we employed a simple and rapid in vitro DSB end joining assay based on fluorescent detection of repair products. Normal and cancer cells were able to repair DNA damage caused by restriction endonucleases, but the efficiency of the end joining was dependent on the type of cells and the structure of DNA ends. K562 cells displayed decreased NHEJ activity in comparison to normal cells for 5' complementary DNA overhang. For blunt-ended DNA there was no significant difference in end joining activity. Both kinds of cells were found about 10-fold more efficient for joining DNA substrates with compatible 5' overhangs than those with blunt ends. Our recent findings have shown that stimulation of DNA repair could be involved in the drug resistance of BCR/ABL-positive cells in anticancer therapy. For the first time the role of STI571 was investigated, a specific inhibitor of BCR/ABL oncogenic protein approved for leukemia treatment in the NHEJ pathway. Surprisingly, STI571 did not change the response of BCR/ABL positive K562 cells in terms of NHEJ for both complementary and blunt ends. Our results suggest that the various responses of the cells to DNA damage via NHEJ can be correlated with the differences in the genetic constitution of human normal and cancer cells. However, the role of NHEJ in anticancer drug resistance in BCR/ABL-positive cells is questionable. PMID- 16042352 TI - In vitro micropropagation of Boswellia ovalifoliolata. AB - A protocol for micropropagation of Boswellia ovalifoliolata Bal & Henry (Burseraceae) was developed using cotyledonary nodal explant on Murashige and Skoog modified medium (MS). A comparative study of micropropagation with 6 benzyladenine, kinetin and thidiazuron along with 1-naphthalene acetic acid (0.054 microM) was conducted. The highest shoot multiplication (7.1 +/- 0.2 shoots per node) was achieved in 50 d on MS supplemented with thidiazuron (2.72 microM). Excised shoot cuttings of 3.0 cm were placed on the MS basal medium supplemented with indole-3-acetic acid and indole-3-butyric acid alone and in combinations for rooting. Activated charcoal (100 mg l(-1)) and polyvinylpyrrolidone (40 mg l(-1)) were added to the medium to prevent browning of cultures. The regenerated plantlets have been successfully acclimatized and transferred to soil. PMID- 16042353 TI - Linum mucronatum: organ to organ lignan variations. AB - The percentage of podophyllotoxin (PTOX) and its congener lignans were measured by HPLC in Linum mucronatum ssp. mucronatum (Linaceae) fresh plant organs. The highest amounts of PTOX (0.595 +/- 0.060% g/g dry wt) and 6 methoxypodophyllotoxin (MPTOX) (1.491 +/- 0.125% g/g dry wt) were found in the plant sexual organs. Whereas, the highest levels of beta-peltatin, 5'-demethoxy MPTOX and yatein were found in not developed buds, petals and sepals, respectively. PMID- 16042354 TI - The epidemiology of partialandrogen deficiency in aging men (PADAM). AB - The quest for eternal youth has been prevalent in civilised societies in many cultures for many centuries. Preventing or deferring the disabilities and morbidities associated with aging through judicious pharmacotherapy has become a particularly relevant healthcare target with the rapid and relentless global demographic shift towards an increasingly elderly population in the 21th century. Aging men commonly loose muscle, become frail, have impaired sexual and cognitive functions, low mood, develop osteopenia and/or osteoporosis with increased risk for fractures and gain visceral fat which predisposes to diabetes, dyslipidaemia, and ischemic heart disease. These alterations in body function are reminiscent of states of androgen deficiency in younger patients. Indeed, aging is associated with a progressive age-related but variable decline in sex hormones. This condition has been named partial androgen deficiency in aging men (PADAM) and consists in a gradual decline in sex hormone levels over years resulting in physical and psychological changes as depression, impotence, decreased sex drive, loss of muscle tone or strength and lethargy. In this review we have tried to give a real identity to PADAM and quantify its entitiy, using the power of the epidemiology. PMID- 16042355 TI - Risk factors for androgen decline in older males: lifestyle, chronic diseases and drugs. AB - The aging process in men is accompanied by a progressive decline in serum testosterone levels. In addition to chronological aging, the various illnesses occurring in mid- to late-adult life and the medications used to treat them further contribute to lowering circulating testosterone levels. Any acute severe illness produces hypogonadotrophic hypogonadism; but also chronic illnesses, such as diabetes, cardiovascular disease and hypertension, lifestyle habits, such as tobacco and alcohol intake, and nutritional factors, from malnutrition to obesity, account for and accentuate the age-related decline in serum androgen levels. These issues are reviewed in a clinical perspective. PMID- 16042356 TI - Late-onset hypogonadism in the aging male (LOH): definition, diagnostic and clinical aspects. AB - Late-onset hypogonadism (LOH) is defined by reduced serum testosterone levels (either total testosterone or free testosterone) and the careful exclusion of any form of classical hypogonadism. When the androgen decline associated with advancing age causes detrimental physiological and mental effects, the syndrome is known as symptomatic LOH (SLOH). A detailed medical history and physical examination are the bases of the diagnosis, and should always precede any biochemical investigations. A general screening of men above a certain age for testosterone deficiency is not feasible. Questionnaires may assist in identifying men who suffer from LOH. Common clinical symptoms of SLOH are lethargy, fatigue, decreased sense of well-being, reduced physical and mental activity, diminished libido, increased sweating, depressive mood, reduced muscle and bone mass or even osteoporosis, erectile dysfunction, and mild anemia. When clinical symptoms are present, the laboratory work-up should focus on total testosterone serum levels. Total testosterone levels <200 ng/dl indicate hypogonadism. In cases of testosterone levels between 200 and 400 ng/dl, measurement should be repeated and supplemented by determination of free testosterone, either by appropriate laboratory methods or the calculation of free testosterone index. In case of very low testosterone levels, classical secondary hypogonadism needs to be considered and excluded. For the safety reasons to exclude contraindications of therapy with androgens, and for follow-up investigations during therapy prostate-specific antigen (PSA), hemoglobin and hematocrit are of interest. PMID- 16042357 TI - Hormonal cut-offs of partial androgen deficiency: a survey of androgen assays. AB - While the typical symptomatology of severe hypogonadism in young adults permits the clinical diagnosis of hypoandrogenism, the diagnosis of partial androgen deficiency of the aging male (PADAM), as occurs relatively frequently in elderly males, is much more difficult. This is due to its clinical picture being subtle and aspecific, and to the fact that both clinical and biochemical evidence of androgen deficiency are required for the diagnosis. In the absence of a practical, clinical useful parameter of androgen activity, we have to rely upon bio-active plasma testosterone levels. Although the requirements of androgens in elderly males may be different from those in young, healthy adults, the lower limit of levels observed in the latter is generally used to define biochemical hypoandrogenism. Based on these data, we consider 11 nmol/l of testosterone, 0.225 nmol/l of free testosterone (FT) and 5.3 nmol/l of biotestosterone (bio-T) as the lower normal limits. As even when using the same kits, values for plasma testosterone and SHBG may differ significantly between laboratories, each laboratory should define its own normal values. As to the methodology, neither direct measurement of free testosterone by analog assay, nor the FT index (T/ SHB) can be recommended, only values obtained by dialysis, ammoniumsulfate precipitation or calculation yielding reliable estimates of androgen bio activity. Dialysis and ammoniumsulfate precipitation are however work intensive and not widely used. PMID- 16042358 TI - Effects of androgens on the cardiovascular system. AB - The evidence that men have a greater incidence of coronary artery disease than women of similar age, together with the fact that android fat distribution is associated with a greater incidence of coronary heart disease, have suggested that high testosterone levels are associated with an increased risk for coronary artery disease. The possible causal role of androgens in the development of cardiovascular disease has not been proven and, to date, there are no epidemiological and pathophysiological evidences to support that an hyper androgenic state or androgen replacement is associated with cardiovascular disease in both sexes. Clinical studies have suggested that physiological testosterone supplementation in ageing males has a positive effect upon lipid profile. Additional potential protective cardiovascular effects of androgens may be related to their effect upon endothelial function and vasomotor tone. Few data are currently available on the correlation between plasma testosterone levels and coronary artery disease in men. Cross-sectional studies reported either reduced or similar plasma testosterone levels and/or androgens in patients with coronary artery disease as compared to controls without cardiovascular symptoms. Epidemiological studies addressing the importance of androgen levels upon cardiovascular mortality and morbidity have gathered inconclusive results. Prospective studies found no significant association between plasma testosterone and cardiac events in both sexes, while most cross-sectional studies have repetitively found an association between hypotestosteronemia and cardiovascular morbidity. In conclusion, androgens in general and testosterone in particular may have some protective effects on the cardiovascular system through their metabolic and direct effects upon human vasculature. PMID- 16042359 TI - Testosterone and sexual activity. AB - Male sexual activity is characterized by a synchronization of sexual desire arising in the brain and its transmission to the periphery, resulting in penile tumescence necessary for sexual intercourse. Testosterone (T) has been claimed for so long as a pivotal hormone in regulating male sexual function, acting both at central and peripheral level. We believe that T is indeed the main synchronizer of sexual activity regulating libido and enzymes as nitric oxide synthase (NOS) and phosphodiesterase type 5 (PDE5), which are crucial for the erectile process. In fact, NOS increases cyclic guanosine monophosphate (cGMP) levels, while PDE5 reduces it. Because T positively controls both the initiation and the end of the penile erection, its net effect on erection is null. In fact, penile erections are often present even without T. The main action of T is to timely adjust the erectile process as a function of sexual desire, therefore finalizing erections to sex. PMID- 16042360 TI - New achievement and novel therapeutic applications of PDE5 inhibithors in older males. AB - Erectile dysfunction (ED) increases in prevalence and severity because of aging processes and related organic, iatrogenic and social problems. Decline of testosterone (T) levels is observed with age and also in illnesses with a common basis of endothelial damage. The T deficiency may lead to decreased energy, mood depression, reduction of sexual desire, but no correlation has been reported between T level and severity of ED, which is mainly a neurovascular disease. In facts, inhibition of phosphodiesterase type 5 (PDE5) isoenzyme with sildenafil, tadalafil and vardenafil enhances vasodilatation in the corpus cavernosum and subsequent penile erection. Absolute pharmacological potency of PDE5 inhibitors is similar and non-selectivity defines the side-effects profile, while their elimination half-life explains not only the different duration of action, but also short and long-term tolerability. Efficacy of PDE5 inhibitors in younger patients is greater in respect to older subjects because of associated pathologies and the decline in hypothalamic-pituitary-gonadal function. T is essential in erectile function, controlling the expression and activity of PDE5 and therefore, androgen supplementation improves therapeutic response to PDE5 inhibitors in hypogonadal subjects. Since sexual behavior is a complex interplay of physical, psychological, and social factors, the possible effect of these drugs on androgen levels and brain function need to be deeply investigated. The ubiquitarious distribution of PDE5 and the availability of selective inhibitory molecules foster newer studies in the treatment of heart failure, pulmonary hypertension, inflammation, and depression. This new progress is certainly contributing to a better medical approach to sexuality and quality of life in aging people. PMID- 16042361 TI - Androgens and fertility. AB - Androgens play a pivotal role in the development of the male reproductive tract. The spermatogenesis requires high levels of intratesticular testosterone secreted by the Leydig cells. Testosterone exerts its action through the androgen receptor (AR), which is located both in the cytoplasm and in the nucleus of cells in the target tissue. Severe defects of the AR may result in abnormal male sexual development, while more subtle modifications can be a potential cause of male infertility. Low circulating levels of testosterone can be found in 20-30% of infertile men, but administration of testosterone or gonadotropins does not result in improved sperm production. Abuse of anabolic steroids is a frequent cause of male infertility, and substances such as endocrine disruptors can alter male fertility through an anti androgenic action. PMID- 16042362 TI - Androgens and body composition in the aging male. AB - The relevant age-related changes in male body composition are mainly related to the progressive decrease in the level of circulating anabolic hormones, among which testosterone (T) is rather important. Its decline, between the ages of 35 and 75, is associated to a loss of muscle mass and fibers number, a doubling of fat mass and a decrease in bone mineral density by 0.3% per yr after age 35; thus the relationship between age-related changes in body composition and T bioactivity reflects an important endocrine aspect of the aging male. The assessment of human body composition and in particular the evaluation of fat tissue and its distribution, is currently standardized by the use of dual-energy x-ray absorpiometry (DXA). In the present paper we review the mechanisms through which testosterone may inhibit adipogenesis, restore the myogenic programme enhancing the protein turnover at muscle level and maintain bone mineral density in elderly men. PMID- 16042363 TI - Testosterone, aging and quality of life. AB - Testosterone levels decline over the lifespan. Many symptoms of hypogonadism are similar to age-related changes in older males. A small number of studies have suggested that some of these symptoms may be reversed by testosterone. Among them, one trial has suggested that decline in bioavailable testosterone may be related to the development of functional decline. Testosterone replacement during rehabilitation may improve functional outcomes and plays an important role in the maintenance of sexual related quality of life. The overall improvement in well being and/or health-related quality of life in older males needs to be determined with large placebo-controlled trials. This is particularly important in view of the substantial placebo effect on aging symptomatology. PMID- 16042364 TI - Androgenic-anabolic steroids abuse in males. AB - Androgenic-anabolic steroids (AAS) is an official definition for all male sex steroid hormones, their synthetic derivatives and their active metabolites. AAS are drugs with specific therapeutic indications, yet they are popularly known because of their worldwide non-therapeutic use in a large number of healthy individuals. Doping with AAS has become so widespread in athletics that it affects the outcome of sports contests. Furthermore, AAS non-therapeutic use is increasing particularly among adolescents and females, becoming one of the main causes of iatrogenic diseases due to drug abuse. All physicians must be aware about the large diffusion and side effects related to AAS non-therapeutic use, in order to discover clinical signs of AAS abuse and/or to start adequate preventive and/or therapeutic actions. PMID- 16042365 TI - Associated hormonal declines in aging: DHEAS. AB - DHEA and its sulfate prohormone DHEAS are the most abundant circulating adrenal steroid hormones in humans. DHEA exerts its actions on peripheral target tissues either indirectly, following its conversion to androgens, estrogens or both, or directly, as a steroid hormone interacting with either a nuclear or a membrane receptor. In humans, DHEA shows a characteristic pattern of secretion throughout life. Serum DHEA concentrations decline with advancing age and vary with gender, ethnicity, and environmental factors. Epidemiological studies show an inverse relationship between plasma DHEA(S) levels in men and age-related illnesses, including cardiovascular and metabolic diseases, immune disorders, malignancies, and neurological dysfunction. This has generated great interest on the putative role of DHEA in age-associated illnesses. Administration of DHEA to rats and mice reduces visceral fat accumulation, and improves insulin resistance in experimental models of diet-induced obesity and/or Type 2 diabetes. In addition, recent studies in vitro have shown that DHEA has the capacity to improve endothelial function by increasing nitric oxide (NO) synthesis. Replacement of DHEA in patients with adrenal insufficiency has been shown to exert beneficial effects on well-being, mood, and sexuality. By contrast, in healthy individuals, the physiological age-associated decline in circulating DHEA(S) per se does not justify DHEA supplementation, since the effects of this hormone on metabolic abnormalities, endothelial function in vivo, and cardiovascular events are contradictory. However, these results do not exclude the possibility that DHEA treatment may prove beneficial in specific subgroups of elderly subjects. PMID- 16042366 TI - Somatopause reflects age-related changes in the neural control of GH/IGF-I axis. AB - This paper will focus on the hypofunction of GH/IGF-I axis in aging, as the most impressive example of decreased activity as function of age-related changes in the neural control of somatotroph cells. GH secretion undergoes clear age-related variations that are generally mirrored by IGF-I levels, the best marker of GH status. Given the well known positive influence of GH/IGF-I on body composition, structure functions and metabolism, this paper discusses the potential clinical implications, also taking into account evidence showing that, at least in animals, deficiency in GH/IGF-I is somewhat associated to prolonged life. Although somatopause is likely to contribute to age-related changes in body composition, structure functions and metabolism, we are now in front of the paradox of lifelong GH/IGF-I deficiency or resistance resulting in prolonged life expectancy and GH replacement at advanced age, probably exerting anti-aging effects. This evidence questions whether GH deficiency is or not a beneficial adaptation to aging. By answering this question one is not simply finding new phylosophical paradigm but also the rational basis for anti-aging drug interventions. PMID- 16042367 TI - Associated hormonal decline in aging: is there a role for GH therapy in aging men? AB - Aging is associated with decline in the somatotroph axis, a decline that has been considered to cause many of the catabolic sequelae of normal aging. Consequently, IGF-I levels decline progressively: this mainly reflects the impaired GH secretion, but decline in gonadal sex steroids and malnutrition also play a role. Decreases in GH secretion may partially explain the age-related changes in metabolism, bones, muscles, cardiovascular system, central nervous system, the immune system and sense of well-being. Normal aging and GH deficiency (GHD) share several clinical signs and symptoms: the endocrine pattern of aging is, however, distinct from the decrease of GH and/or IGF-I levels associated with hypopituitarism. Owing to clinical similarities between aging and GHD, the relative GH insufficiency of elderly subjects has been postulated as one important factor contributing to their frailty. However, it is currently unclear whether treatment with exogenous GH, or IGF-I can retard or reverse age-related changes in body structure and function. Several studies enrolling healthy elderly subjects showed that GH treatment was followed by increase of lean mass, decrease of fat mass and improvement of bone turnover, but other studies failed to confirm these data. The treatment with natural or synthetic GH secretagogues, that could represent a physiological approach to restore the GH and IGF-I secretion, requires further investigations. In conclusion, there is not sufficient evidence for a clear therapeutic role of rhGH, IGF-I, and secretagogues in aging. Further studies are needed to evaluate the real benefit of somatotropic treatment in aging men. PMID- 16042368 TI - Pratical aspects of testosterone substitution. AB - Until more data on the efficacy and risks of testosterone (T) treatment of hypogonadal older men become available, the use of T substitution in men in this age group should be carefully considered and benefits vs risks should be discussed with the patient. Generally symptoms of hypogonadism together with low serum T levels are considered indications for treatment. For aging men, short acting preparations may be preferred and levels of serum T are maintained with in the mid adult range. Careful assessment for improvement of symptoms is the essential goal of treatment. Monitoring for manifestations of prostate disease, erythrocytosis and other adverse events of androgen treatment is critical in the management of older men with androgen replacement therapy. PMID- 16042369 TI - Transdermal androgens: pharmacology and applicability to hypogonadal elderly men. AB - Transdermal testosterone (T) gels have proven to be an efficacious means of delivering T to hypogonadal men. Patches, gels and creams have been used in T transdermal formulations. Patches suffered from problems either with adherence or skin irritability while gels have a potential problem of transferability from patients to partner or other family member. At present, gels have been well received by clinicians and users. There is no evidence that transdermal preparations are not suitable for treatment of older hypogonadal men; the rapid off time of both patches and gels may argue allow that transdermal preparations may be a preferential treatment modality for older men with a potential higher risk of developing androgen-responsive prostate cancer. PMID- 16042370 TI - Psychosocial issues of ART in aging male. AB - The role of androgens in human sexuality as regards the mechanism of erection and the pathogenesis of impotence is under debate. In addition, it is difficult to define the psychosocial impact of both hypogonadism and androgen replacement. However, sexual hormones largely influence mood, well-being, and quality of life. For this reason, despite the methodological difficulties of assessment, testosterone replacement has a deep impact on the social, psychological and sexual life of the treated patient. Considering the obvious characteristic of testosterone as an hormone, it appears evident that the endocrinologist is the unique experienced specialist able to diagnose and treat the hypogonadal men, monitoring potential side effects and following the psychosocial issues of androgen therapy. PMID- 16042371 TI - Monitoring androgen replacement therapy: testosterone and prostate safety. AB - The aging of the world population has brought to the forefront of medical practice the diagnosis and treatment of hypogonadism in adult men. There is an increasing interest on the use of testosterone (T) and other androgens to manage men with clinical and biochemical evidence of hypogonadism. Although treatment with T has been used for 70 yr and it is, generally, safe and effective, there are a number of safety issues--ranging from cardiovascular and lipid alterations to hematological changes--that the physician needs to be aware of. Unquestionably, prostate safety constitutes the most important one. No evidence exists that appropriate androgen administration with knowledgeable monitoring carries significant or potentially serious adverse effects on the prostate gland. Men with symptomatic lower urinary obstruction need to be assessed carefully prior to androgen administration. The suspicion of prostate cancer is an absolute contraindication for T use. Recommendations are available for the judicious and safe use of T in aging men. PMID- 16042372 TI - Fixation of the cemented acetabular component in hip arthroplasty. PMID- 16042373 TI - Transpeptidation reactions of a specific substrate catalyzed by the Streptomyces R61 DD-peptidase: the structural basis of acyl acceptor specificity. AB - Bacterial dd-peptidases, the targets of beta-lactam antibiotics, are believed to catalyze d-alanyl-d-alanine carboxypeptidase and transpeptidase reactions in vivo. To date, however, there have been few concerted attempts to explore the kinetic and thermodynamic specificities of the active sites of these enzymes. We have shown that the peptidoglycan-mimetic peptide, glycyl-l-alpha-amino-epsilon pimelyl-d-alanyl-d-alanine, 1, is a very specific and reactive carboxypeptidase substrate of the Streptomyces R61 dd-peptidase [Anderson, J. W., and Pratt, R. F. (2000) Biochemistry 39, 12200-12209]. In the present paper, we explore the transpeptidation reactions of this substrate, where the enzyme catalyzes transfer of the glycyl-l-alpha-amino-epsilon-pimelyl-d-alanyl moiety to amines. These reactions are believed to occur through capture of an acyl-enzyme intermediate by amines rather than water. Experiments show that effective acyl acceptors require a carboxylate group and thus are amino acids and peptides. d(but not l)-amino acids, analogues of the leaving group of 1, are good acceptors. The effectiveness of d-alanine as an acceptor increases with pH, suggesting that the bound and reactive form of an amino acid acceptor is the free amine. Certain glycyl-l(but not d)-amino acids, such as glycyl-l-alanine and glycyl-l-phenylalanine, are also good acceptors. These molecules may resemble the N-terminus of the Streptomyces stem peptides that, presumably, are the acceptors in vivo. The acyl acceptor binding site therefore demonstrates a dual specificity. That d-alanyl-l-alanine shows little activity as an acceptor suggested that, on binding of acceptors to the enzyme, the carboxylate of d-amino acids does not overlap with the peptide carbonyl group of glycyl-l-amino acids. Molecular modeling of transpeptidation tetrahedral intermediates and products demonstrated the likely structural bases for the stereospecificity of the acceptors and the nature of the dual function acceptor binding site. For both groups of acceptors, the terminal carboxylate appeared to be anchored at the active site by interaction with Arg 285 and Thr 299. PMID- 16042374 TI - Transpeptidation reactions of a specific substrate catalyzed by the streptomyces R61 DD-peptidase: characterization of a chromogenic substrate and acyl acceptor design. AB - The Streptomyces R61 dd-peptidase, a functional model for penicillin-binding proteins, catalyzes the hydrolysis and aminolysis of d-alanyl-d-alanine terminating peptides by specific amines. In vivo, this reaction completes bacterial cell wall biosynthesis. For in vitro studies of this enzyme to date, various nonspecific acyl-donor substrates have been employed. Recently, however, a peptidoglycan-mimetic peptide substrate, glycyl-l-alpha-amino-epsilon-pimelyl-d alanyl-d-alanine, has been described that is much more specific for this enzyme. In this paper, we describe the synthesis and kinetic characterization of an analogous thiolester substrate, 3-(N-glycyl-l-cysteinyl)-propanoyl-d-alanyl-d thiolactate, that the enzyme hydrolyzes and aminolyzes very efficiently (k(cat)/K(m) = 1.0 x 10(7) s(-)(1) M(-)(1)). Direct or indirect, by means of a thiol trap, spectrophotometric monitoring of the reactions of this substrate is readily achieved. Deacylation of the enzyme is rate-determining under substrate saturation conditions, and therefore the aminolysis reaction can be directly studied. The results show that d-amino acids and certain Gly-l-Xaa dipeptides and tripeptides may act as acyl acceptors at the active site of the enzyme. d Phenylalanine and Gly-l-Phe were the most effective d-amino acid and dipeptide acceptors, respectively. On the basis of the dual specificity of the active site for acceptors (d-amino acids and Gly-l-Xaa peptides), "dual function" acceptors were designed and synthesized. Two of these, aminomalon-(N-ethyl)amide and aminomalon-(N-phenethyl)amide, were particularly effective. It did seem, however, that the observed rates of reaction of these very effective acceptors may be limited by some common, possibly physical, step. More extended, peptidoglycan like, acceptors were found to be essentially unreactive. The reasons for this counterintuitive behavior are discussed. PMID- 16042375 TI - X-ray structure of the R69D phosphatidylinositol-specific phospholipase C enzyme: insight into the role of calcium and surrounding amino acids in active site geometry and catalysis. AB - Phosphatidylinositol-specific phospholipase Cs (PLCs) are a family of phosphodiesterases that catalyze the cleavage of the P-O bond via transesterification using the internal hydroxyl group of the substrate as a nucleophile, generating the five-membered cyclic inositol phosphate as an intermediate or product. To better understand the role of calcium in the catalytic mechanism of PLCs, we have determined the X-ray crystal structure of an engineered PLC enzyme from Bacillus thuringiensis to 2.1 A resolution. The active site of this enzyme has been altered by substituting the catalytic arginine with an aspartate at position 69 (R69D). This single-amino acid substitution converted a metal-independent, low-molecular weight enzyme into a metal ion-dependent bacterial PLC with an active site architecture similar to that of the larger metal ion-dependent mammalian PLC. The Ca(2+) ion shows a distorted square planar geometry in the active site that allows for efficient substrate binding and transition state stabilization during catalysis. Additional changes in the positions of the catalytic general acid/general base (GA/GB) were also observed, indicating the interrelation of the intricate hydrogen bonding network involved in stabilizing the active site amino acids. The functional information provided by this X-ray structure now allows for a better understanding of the catalytic mechanism, including stereochemical effects and substrate interactions, which facilitates better inhibitor design and sheds light on the possibilities of understanding how protein evolution might have occurred across this enzyme family. PMID- 16042376 TI - The DRY motif as a molecular switch of the human oxytocin receptor. AB - The human oxytocin receptor is known to exhibit promiscuous activity by coupling to both Galpha(q) and Galpha(i) G proteins to activate distinct signaling pathways. A single-amino acid substitution within the highly conserved E/DRY motif at the cytosolic extension of helix 3 [i.e., D136(3.49)N] increased the rate of both basal and agonist-stimulated inositol phosphate (IP(3)) accumulation of the receptor. Furthermore, like for a typical constitutively active receptor, the partial agonist arginine vasopressin behaved as a full agonist for the D136(3.49)N mutant. Subsequently, both oxytocin and arginine vasopressin showed an increased potency in stimulating IP3 accumulation as compared to the wild-type receptor. Very interestingly, our experiments provide strong evidence that the D136(3.49)N mutant inhibits receptor signaling via Galpha(i)-mediated pathways while increasing the activity through the Galpha(q)-mediated pathways. Molecular simulations of the free and OT-bound forms of wild-type OTR and of the D136(3.49)N constitutively active mutant suggest that the receptor portions close to the E/DRY and NPxxY motifs are particularly susceptible to undergoing structural modification in response to activating mutations and agonist binding. Furthermore, computational modeling suggests that the OT-bound form of wild-type OTR is able to explore more states than the OT-bound form of the D136(3.49)N constitutively active mutant, consistent with its G protein promiscuity. Taken together, these observations emphasize the important role of the E/DRY motif not only in receptor activation but also in the promiscuity of G protein coupling. Knowledge of the mechanism of selective G protein coupling could aid drug discovery efforts to identify signaling specific therapies. PMID- 16042377 TI - Molecular-level investigation of the structure, transformation, and bioactivity of single living fission yeast cells by time- and space-resolved Raman spectroscopy. AB - The structure, transformation, and bioactivity of single living Schizosaccharomyces pombe cells at the molecular level have been studied in vivo by time- and space-resolved Raman spectroscopy. A time resolution of 100 s and a space resolution of 250 nm have been achieved with the use of a confocal Raman microspectrometer. The space-resolved Raman spectra of living S. pombe cells at different cell cycle stages were recorded in an effort to elucidate the molecular compositions of organelles, including nuclei, cytoplasm, mitochondria, and septa. The time- and space-resolved measurement of the central part of a dividing yeast cell showed continuous spectral evolution from that of the nucleus to those of the cytoplasm and mitochondria and finally to that of the septum, in accordance with the transformation during the cell cycle. A strong Raman band was observed at 1602 cm(-)(1) only when cells were under good nutrient conditions. The effect of a respiration inhibitor, KCN, on a living yeast cell was studied by measuring the Raman spectra of its mitochondria. A sudden disappearance of the 1602 cm( )(1) band followed by the change in the shape and intensity of the phospholipid bands was observed, indicating a strong relationship between the cell activity and the intensity of this band. We therefore call this band "the Raman spectroscopic signature of life". The Raman mapping of a living yeast cell was also carried out. Not only the distributions of molecular species but also those of active mitochondria in the cell were successfully visualized in vivo. PMID- 16042378 TI - Araneoid egg case silk: a fibroin with novel ensemble repeat units from the black widow spider, Latrodectus hesperus. AB - Araneoid spiders use specialized abdominal glands to manufacture up to seven different protein-based silks/glues that have diverse physical properties. The fibroin sequences that encode egg case fibers (cover silk for the egg case sac) and the secondary structure of these threads have not been previously determined. In this study, MALDI tandem TOF mass spectrometry (MS/MS) and reverse genetics were used to isolate the first egg case fibroin, named tubuliform spidroin 1 (TuSp1), from the black widow spider, Latrodectus hesperus. Real-time quantitative PCR analysis demonstrates TuSp1 is selectively expressed in the tubuliform gland. Analysis of the amino acid composition of raw egg case silk closely aligns with the predicted amino acid composition from the primary sequence of TuSp1, which supports the assertion that TuSp1 represents a major component of egg case fibers. TuSp1 is composed of highly homogeneous repeats that are 184 amino acids in length. The long stretches of polyalanine and glycine alanine subrepeats, which account for the crystalline regions of minor ampullate and major ampullate fibers, are very poorly represented in TuSp1. However, polyserine blocks and short polyalanine stretches were highly iterated within the primary sequence, and (13)C NMR spectroscopy demonstrated that the majority of alanine was found in a beta-sheet structure in post-spun egg case silk. The TuSp1 repeat unit does not display substantial sequence similarity to any previously described fibroin genes or proteins, suggesting that TuSp1 is a highly divergent member of the spider silk gene family. PMID- 16042379 TI - Characterization of conformational changes coupled to ligand photodissociation from the heme binding domain of FixL. AB - Using transient absorption spectroscopy and photoacoustic calorimetry (PAC), we have characterized carbon monoxide photodissociation and rebinding to two forms of the heme domain of Bradyrhizobium japonicum FixL. Transient absorption results for the complete heme domain (FixL residues 140-270) and a truncated heme domain (missing 11 residues on the N-teminal end and 14 amino acid residues on the C terminal end of the full length heme domain) show similar rates for ligand rebinding to the five-coordinate heme domain and the absence of any transient intermediate on a microsecond time scale. Results from PAC studies show that both the truncated and complete heme domains undergo a contraction upon ligand photolysis. In addition, CO photolysis from the complete heme domain gives rise to an intermediate with a lifetime of approximately 150 ns which is absent in the truncated heme domain. We attribute the 150 ns phase to ligand release to the solvent which may be accelerated in the case of the truncated domain. The initial contraction is attributed to changes in the charge distribution due to reorganization of the surface salt bridge formed between Glu182 and Arg227 or possibly to reorientation of Arg206. Changes in the charge distribution may play an important role in communication between the sensor domain and the regulatory domain and thus may be part of the signal transduction pathway. PMID- 16042380 TI - New class of bacterial membrane oxidoreductases. AB - A new class of bacterial multisubunit membrane-bound electron-transfer complexes has been identified based on biochemical and bioinformatic data. It contains subunits homologous to the three-subunit molybdopterin oxidoreductases and four additional subunits, two of which are c-type cytochromes. The complex was purified from the filamentous anoxygenic phototrophic bacterium Chloroflexus aurantiacus, and putative operons for similar complexes were identified in a wide range of bacteria. In most cases, the presence of the new complex is anticorrelated with the cytochrome bc or bf electron-transfer complex, suggesting that it replaces it functionally. This appears to be a widespread yet previously unrecognized protein complex involved in energy metabolism in bacteria. PMID- 16042381 TI - Human carbonic anhydrase III: structural and kinetic study of catalysis and proton transfer. AB - The residue phenylalanine 198 (Phe 198) is a prominent cause of the lower activity of human carbonic anhydrase III (HCA III) compared with HCA II and other isozymes which have leucine at this site. We report the crystal structures of HCA III and the site-directed mutant F198L HCA III, both at 2.1 A resolution, and the enhancement of catalytic activity by exogenous proton donors containing imidazole rings. Both enzymes had a hexahistidine extension at the carboxy-terminal end, used to aid in purification, that was ordered in the crystal structures bound in the active site cavity of an adjacent symmetry-related enzyme. This observation allowed us to comment on a number of possible binding sites for imidazole and derivatives as exogenous proton donors/acceptors in catalysis by HCA III. Kinetic and structural evidence indicates that the phenyl side chain of Phe 198 in HCA III, about 5 A from the zinc, is a steric constriction in the active site, may cause altered interactions at the zinc-bound solvent, and is a binding site for the activation of catalysis by histidylhistidine. This suggests that sites of activation of the proton-transfer pathway in carbonic anhydrase are closer to the zinc than considered in previous studies. PMID- 16042382 TI - Snapshots of a dynamic folding nucleus in zinc-substituted Pseudomonas aeruginosa azurin. AB - Zinc-substituted Pseudomonas aeruginosa azurin folds in two-state equilibrium and kinetic reactions. In the unfolded state, the zinc ion remains bound to the unfolded polypeptide via two native-state ligands (His117 and Cys112). The significantly curved Chevron plot for zinc-substituted azurin was earlier ascribed to movement of the folding-transition state. At low concentrations of denaturant, the transition state occurs early in the folding reaction (low Tanford beta-value), whereas at high-denaturant concentration, it moves closer to the native structure (high Tanford beta-value). Here, we use this movement to track the formation and growth of zinc-substituted azurin's folding nucleus with atomic resolution using protein engineering. The average phi (phi) value for 17 positions (covering all secondary-structure elements) goes from 0.25 in 0 M GuHCl (beta approximately 0.46) to 0.76 in 4 M GuHCl (beta approximately 0.86); a phi value of 1 or 0 indicates native-like or unfolded-like interactions, respectively. Analysis of individual phi-values reveals a delocalized nucleus where structure condenses around a leading density centered on Leu50 in the core. The diffuse moving transition state for zinc-substituted azurin is in sharp contrast to the fixed polarized folding nucleus observed for apo-azurin. The dramatic difference in apparent kinetic behavior for the two forms of azurin can be rationalized as a minor alteration on a common free-energy profile that exhibits a broad activation barrier. PMID- 16042383 TI - Functional mapping of charged residues of the 82-116 sequence in factor Xa: evidence that lysine 96 is a factor Va independent recognition site for prothrombin in the prothrombinase complex. AB - It has been hypothesized that two antiparallel structures comprised of residues 82-91 and 102-116 in factor Xa (fXa) may harbor a factor Va- (fVa-) dependent prothrombin recognition site in the prothrombinase complex. There are 11 charged residues in the 82-116 loop of human fXa (Glu-84, Glu-86, Lys-90, Arg-93, Lys-96, Glu-97, Asp-100, Asp-102, Arg-107, Lys-109, and Arg-115). With the exception of Glu-84, which did not express, and Asp-102, which is a catalytic residue, we expressed the Ala substitution mutants of all other residues and evaluated their proteolytic and amidolytic activities in both the absence and presence of fVa. K96A and K109A activated prothrombin with 5-10-fold impaired catalytic efficiency in the absence of fVa. All mutants, however, exhibited normal activity toward the substrate in the presence of fVa. K109A also exhibited impaired amidolytic activity and affinity for Na(+); however, both fVa and higher Na(+) restored the catalytic defect caused by the mutation. Analysis of the X-ray crystal structure of fXa indicated that Glu-84 may interact by a salt bridge with Lys-109, explaining the lack of expression of E84A and the lower activity of K109A in the absence of fVa. These results suggest that none of the residues under study is a fVa-dependent recognition site for prothrombin in the prothrombinase complex; however, Lys-96 is a recognition site for the substrate independent of the cofactor. Moreover, the 82-116 loop is energetically linked to fVa and Na(+) binding sites of the protease. PMID- 16042384 TI - Crystal structure of Methanobacterium thermoautotrophicum phosphoribosyl-AMP cyclohydrolase HisI. AB - The metabolic pathway for histidine biosynthesis is interesting from an evolutionary perspective because of the diversity of gene organizations and protein structures involved. Hydrolysis of phosphoribosyl-AMP, the third step in the histidine biosynthetic pathway, is carried out by PR-AMP cyclohydrolase, the product of the hisI gene. The three-dimensional structure of PR-AMP cyclohydrolase from Methanobacterium thermoautotrophicum was solved and refined to 1.7 A resolution. The enzyme is a homodimer. The position of the Zn(2+) binding site that is essential for catalysis was inferred from the positions of bound Cd(2+) ions, which were part of the crystallization medium. These metal binding sites include three cysteine ligands, two from one monomer and the third from the second monomer. The enzyme remains active when Cd(2+) is substituted for Zn(2+). The likely binding site for Mg(2+), also necessary for activity in a homologous cyclohydrolase, was also inferred from Cd(2+) positions and is comprised of aspartic acid side chains. The putative substrate-binding cleft is formed at the interface between the two monomers of the dimer. This fact, combined with the localization of the Zn(2+)-binding site, indicates that the enzyme is an obligate dimer. PMID- 16042386 TI - Transient interaction with nanoparticles "freezes" a protein in an ensemble of metastable near-native conformations. AB - It is well-known that adsorption of proteins on interfaces often induces substantial alterations of the protein structure. However, very little is known about whether these conformational changes have any consequence for the protein conformation after desorption from the interface. To investigate this matter, we have selected a protein-particle system in which the enzyme human carbonic anhydrase I (HCAI) alternates between the adsorbed and free state upon interaction with the silica nanoparticles. High-resolution NMR analysis of the protein with the particles present in the sample shows a spectrum that indicates a molten globular-like structure. Removal of particles results in refolding of virtually all HCAI molecules to a fully active form. However, the two-dimensional NMR analysis shows that refolding does not result in a single well-defined protein structure but rather provides an ensemble of protein molecules with near native conformations. A detailed comparative chemical shift analysis of 108 amide signals in (1)H-(15)N HSQC spectra of native and desorbed HCAI reveals that the most profound effects are located at beta-strands in the center of the molecule. The observation of very slow H-D exchange in the central beta-strands of HCAI [Kjellsson, A., Sethson, I., and Jonsson, B. H. (2003) Biochemistry 42, 363-374] in conjunction with our results indicates that the kinetic barriers for conformational rearrangements in the central core of the protein are low in the presence of nanoparticles but are very high under native conditions. PMID- 16042385 TI - Structure of the 1,4-bis(2'-deoxyadenosin-N6-yl)-2R,3R-butanediol cross-link arising from alkylation of the human N-ras codon 61 by butadiene diepoxide. AB - The solution structure of the 1,4-bis(2'-deoxyadenosin-N(6)-yl)-2R,3R-butanediol cross-link arising from N(6)-dA alkylation of nearest-neighbor adenines by butadiene diepoxide (BDO(2)) was determined in the oligodeoxynucleotide 5' d(CGGACXYGAAG)-3'.5'-d(CTTCTTGTCCG)-3'. This oligodeoxynucleotide contained codon 61 (underlined) of the human N-ras protooncogene. The cross-link was accommodated in the major groove of duplex DNA. At the 5'-side of the cross-link there was a break in Watson-Crick base pairing at base pair X(6).T(17), whereas at the 3' side of the cross-link at base pair Y(7).T(16), base pairing was intact. Molecular dynamics calculations carried out using a simulated annealing protocol, and restrained by a combination of 338 interproton distance restraints obtained from (1)H NOESY data and 151 torsion angle restraints obtained from (1)H and (31)P COSY data, yielded ensembles of structures with good convergence. Helicoidal analysis indicated an increase in base pair opening at base pair X(6).T(17), accompanied by a shift in the phosphodiester backbone torsion angle beta P5'-O5'-C5'-C4' at nucleotide X(6). The rMD calculations predicted that the DNA helix was not significantly bent by the presence of the four-carbon cross link. This was corroborated by gel mobility assays of multimers containing nonhydroxylated four-carbon N(6),N(6)-dA cross-links, which did not predict DNA bending. The rMD calculations suggested the presence of hydrogen bonding between the hydroxyl group located on the beta-carbon of the four-carbon cross-link and T(17) O(4), which perhaps stabilized the base pair opening at X(6).T(17) and protected the T(17) imino proton from solvent exchange. The opening of base pair X(6).T(17) altered base stacking patterns at the cross-link site and induced slight unwinding of the DNA duplex. The structural data are interpreted in terms of biochemical data suggesting that this cross-link is bypassed by a variety of DNA polymerases, yet is significantly mutagenic [Kanuri, M., Nechev, L. V., Tamura, P. J., Harris, C. M., Harris, T. M., and Lloyd, R. S. (2002) Chem. Res. Toxicol. 15, 1572-1580]. PMID- 16042387 TI - Probing PrPSc structure using chemical cross-linking and mass spectrometry: evidence of the proximity of Gly90 amino termini in the PrP 27-30 aggregate. AB - Elucidation of the structure of PrP(Sc) continues to be one of the most important and difficult challenges in prion research. This task, essential for gaining an understanding of the basis of prion infectivity, has been hampered by the insoluble, aggregated nature of this molecule. We used a combination of chemical cross-linking, proteolytic digestion, and mass spectrometry (MALDI-TOF and nanoLC ESI-QqTOF), in an attempt to gain structural information about PrP 27-30 purified from the brains of Syrian hamsters infected with scrapie. The rationale of this approach is to identify pairs of specific amino acid residues that are close enough to each other to react with a bifunctional reagent of a given chain length. We cross-linked PrP 27-30 with the amino-specific reagent bis(sulfosuccinimidyl) suberate (BS(3)), obtaining dimers, trimers, and higher order oligomers that were separated by SDS-PAGE. In-gel digestion followed by mass spectrometric analysis showed that BS(3) reacted preferentially with Gly90. A cross-link involving two Gly90 amino termini was found in cross-linked PrP 27 30 dimers, but not in intramolecularly cross-linked monomers or control samples. This observation indicates the spatial proximity of Gly90 amino termini in PrP 27 30 fibrils. The Gly90-Gly90 cross-link is consistent with a recent model of PrP 27-30, based on electron crystallographic data, featuring a fiber composed of stacked trimers of PrP monomers; specifically, it is compatible with cross linking of monomers stacked vertically along the fiber axis but not those adjacent to each other horizontally in the trimeric building block. Our results constitute the first measured distance constraint in PrP(Sc). PMID- 16042388 TI - Mechanism of penetration of Antp(43-58) into membrane bilayers. AB - Antp(43-58) is one of many peptides with basic and aromatic residues capable of crossing cell membranes efficiently in a receptor-independent manner. The basic aromatic motif is responsible for peptide binding to the negatively charged surface of membrane bilayers. However, the mechanism of membrane penetration is unclear. We use high-resolution (1)H solution NMR methods to establish the location of the Antp(43-58) peptide bound to membrane bicelles composed of DMPC, DMPG, and DHPC, and compare it to the location of an Antp(43-58) variant which is not able to cross cell membranes. Two critical tryptophans are substituted with phenylalanine in this variant (W48F and W56F). Additional (31)P and (2)H NMR measurements of membrane bicelles are used to probe the changes in orientation of the lipid headgroups and the changes in the mobility or segmental order of the lipid acyl chains upon peptide binding. We find that Trp48 and Trp56 of Antp(43 58) insert into the hydrophobic core of the membrane and that this induces a change in the orientation of the negatively charged DMPG headgroups. The depth of insertion and the change in lipid orientation are concentration-dependent and argue for an electroporation-like mechanism for membrane penetration. PMID- 16042390 TI - Role of the C-terminus of the high-conductance calcium-activated potassium channel in channel structure and function. AB - The role of ion channels in cell physiology is regulated by processes occurring after protein biosynthesis, which are critical for both channel function and targeting of channels to appropriate cell compartments. Here we apply biochemical and electrophysiological methods to investigate the role of the high-conductance, calcium-activated potassium (Maxi-K) channel C-terminal domain in channel tetramerization, association with the beta1 subunit, trafficking of the channel complex to the cell surface, and channel function. No evidence for channel tetramerization, cell surface expression, or function was observed with Maxi K(1)(-)(323), a construct truncated three residues after the S(6) transmembrane domain. However, Maxi-K(1)(-)(343) and Maxi-K(1)(-)(441) are able to form tetramers and to associate with the beta1 subunit. Maxi-K(1)(-)(343)-beta1 and Maxi-K(1)(-)(441)-beta1 complexes are efficiently targeted to the cell surface and cannot be pharmacologically distinguished from full-length channels in binding experiments but do not form functional channels. Maxi-K(1)(-)(651) forms tetramers and associates with beta1; however, the complex is not present at the cell surface, but is retained intracellularly. Maxi-K(1)(-)(651) surface expression and channel function can be fully rescued after coexpression with its C-terminal complement, Maxi-K(652)(-)(1113). However coexpression of Maxi-K(1)( )(343) and Maxi-K(1)(-)(441) with their respective C-terminal complements did not rescue channel function. Together, these data demonstrate that the domain(s) in the Maxi-K channel necessary for formation of tetramers, coassembly with the beta1 subunit, and cell surface expression resides within the S(0)-S(6) linker domain of the protein, and that structural constraints within the gating ring in the C-terminal region can regulate trafficking and function of constructs truncated in this region. PMID- 16042389 TI - PhosphoThr peptide binding globally rigidifies much of the FHA domain from Arabidopsis receptor kinase-associated protein phosphatase. AB - A net increase in the backbone rigidity of the kinase-interacting FHA domain (KI FHA) from the Arabidopsis receptor kinase-associated protein phosphatase (KAPP) accompanies the binding of a phosphoThr peptide from its CLV1 receptor-like kinase partner, according to (15)N NMR relaxation at 11.7 and 14.1 T. All of the loops of free KI-FHA display evidence of nanosecond-scale motions. Many of these same residues have residual dipolar couplings that deviate from structural predictions. Binding of the CLV1 pT868 peptide seems to reduce nanosecond-scale fluctuations of all loops, including half of the residues of recognition loops. Residues important for affinity are found to be rigid, i.e., conserved residues and residues of the subsite for the key pT+3 peptide position. This behavior parallels SH2 and PTB domain recognition of pTyr peptides. PhosphoThr peptide binding increases KI-FHA backbone rigidity (S(2)) of three recognition loops, a loop nearby, seven strands from the beta-sandwich, and a distal loop. Compensating the trend of increased rigidity, binding enhances fast mobility at a few sites in four loops on the periphery of the recognition surface and in two loops on the far side of the beta-sandwich. Line broadening evidence of microsecond- to millisecond-scale fluctuations occurs across the six-stranded beta-sheet and nearby edges of the beta-sandwich; this forms a network connected by packing of interior side chains and H-bonding. A patch of the slowly fluctuating residues coincides with the site of segment-swapped dimerization in crystals of the FHA domain of human Chfr. Phosphopeptide binding introduces microsecond- to millisecond-scale fluctuations to more residues of the long 8/9 recognition loop of KI-FHA. The rigidity of this FHA domain appears to couple as a whole to pThr peptide binding. PMID- 16042391 TI - Blocking effect and crystal structure of natrin toxin, a cysteine-rich secretory protein from Naja atra venom that targets the BKCa channel. AB - Cysteine-rich secretory proteins (CRISPs) are widespread in snake venoms. Some members of these CRISPs recently have been found to block L-type Ca(2+) channels or cyclic nucleotide-gated ion (CNG) channels. Here, natrin purified from Naja atra venom, a member of the CRISP family, can induce a further contractile response in the endothelium-denuded thoracic aorta of mouse which has been contracted by a high-K(+) solution. Further experiments show it can block the high-conductance calcium-activated potassium (BK(Ca)) channel in a concentration dependent manner with an IC(50) of 34.4 nM and a Hill coefficient of 1.02, which suggests that only a single natrin molecule is required to bind an ion channel to block BK(Ca) current. The crystal structure of natrin displaying two domains in tandem shows its cysteine-rich domain (CRD) has relatively independent flexibility, especially for the C-terminal long loop (loop I) of CRD to participate in the interface of two domains. On the basis of previous studies of CNG channel and L-Ca(2+) channel blockers, and the sequence and structural comparison of natrin and stecrisp, the deviation of the vital loop I of CRD is suggested to contribute to different effects of some CRISPs in protein-protein interaction. PMID- 16042392 TI - Solution structure of the peptidoglycan binding domain of Bacillus subtilis cell wall lytic enzyme CwlC: characterization of the sporulation-related repeats by NMR. AB - Bacillus subtilis CwlC is a cell wall lytic N-acetylmuramoyl-l-alanine amidase that plays an important role in mother-cell lysis during sporulation. The enzyme consists of an N-terminal catalytic domain with C-terminal tandem repeats. The repeats [repeat 1 (residues 184-219) and repeat 2 (residues 220-255)] are termed CwlCr. We report on the solution structure of CwlCr as determined by multidimensional NMR, including the use of 36 (h3)J(NC)'-derived hydrogen bond restraints and 64 residual (1)D(NH) dipolar couplings. Two tandem repeats fold into a pseudo-2-fold symmetric single-domain structure consisting of a betaalphabetabetaalphabeta-fold containing numerous contacts between the repeats. Hydrophobic residues important for structural integrity are conserved between the repeats, and are located symmetrically. We also present NMR analysis of the circularly permuted repeat mutant of CwlCr. Secondary structure content from the chemical shifts and hydrogen bonds derived from (h3)J(NC)' show that the mutant folds into a structure similar to that of the wild type, suggesting that the repeats are exchangeable. This implies that conserved hydrophobic residues are crucial for maintaining the folding of the repeats. While monitoring the chemical shift perturbations following the addition of digested soluble peptidoglycan fragments, we identified two peptidoglycan interaction sites of CwlCr at the edges of the protein symmetrically, and they are located approximately 28 A from each other. PMID- 16042393 TI - Evidence for a mechanism by which omega-3 polyunsaturated lipids may affect membrane protein function. AB - We have calculated the lateral pressure profile from well-converged, experimentally validated, molecular dynamics simulations of hydrated lipid bilayer membranes containing highly polyunsaturated fatty acids. The three simulations, each 30 ns in length, contain omega-3 fatty acids, omega-6 fatty acids, and a mixture of omega-3 fatty acids and cholesterol and were continued from previously published simulations that demonstrated excellent agreement with a wide variety of experimental measurements. We find that the distribution of lateral stress within the hydrophobic core of the membrane is sensitively dependent on the degree of chain unsaturation and on the presence of cholesterol. Replacing omega-3 fatty acids with omega-6 chains, or incorporating cholesterol into the membrane, shifts the repulsive lateral chain pressure away from the lipid/water interface toward the bilayer interior. This may support a previously proposed mechanism by which lipid composition may affect conformational equilibrium for integral membrane proteins. PMID- 16042394 TI - Formation of an antiparallel, intermolecular coiled coil is associated with in vivo dimerization of osmosensor and osmoprotectant transporter ProP in Escherichia coli. AB - Membrane transporter ProP from Escherichia coli senses extracellular osmolality and responds by mediating the uptake of osmoprotectants such as glycine betaine when osmolality is high. Earlier EPR and NMR studies showed that a peptide replica of the cytoplasmic ProP carboxyl terminus (residues D468-R497) forms a homodimeric, antiparallel, alpha-helical coiled coil in vitro stabilized by electrostatic interactions involving R488. Amino acid replacement R488I disrupted coiled-coil formation by the ProP peptide, elevated the osmolality at which ProP became active, and rendered the osmolality response of ProP transient. In the present study, either E480 or K473 was replaced with cysteine (Cys) in ProP, a Cys-less, fully functional, histidine-tagged ProP variant, to use Cys-specific cross-linking approaches to determine if antiparallel coiled-coil formation and dimerization of the intact protein occur in vivo. The Cys at positions 480 would be closer in an antiparallel dimer than those at positions 473. These replacements did not disrupt coiled-coil formation by the ProP peptide. Partial homodimerization of variant ProP-E480C could be demonstrated in vivo and in membrane preparations via Cys-specific cross-linking with dithiobis(maleimidoethane) or by Cys oxidation to cystine by copper phenanthroline. In contrast, these reagents did not cross-link ProP with Cys at position 133 or 241. Cross-linking of ProP with Cys at position 473 was limited and occurred only if ProP was overexpressed, consistent with an antiparallel orientation of the coiled coil in the intact protein in vivo. Although replacement E480C did not alter transporter activity, replacement K473C reduced the extent and elevated the threshold for osmotic activation. K473 may play a role in ProP structure and function that is not reflected in altered coiled-coil formation by the corresponding peptide. Substitution R488I affected the activities of ProP-(His)(6), ProP-E480C, and ProP-K473C as it affected the activity of ProP. Surprisingly, it did not eliminate cross-linking of Cys at position 480, and it elevated cross-linking at position 473, even when ProP was expressed at physiological levels. This suggested that the R488I substitution may have changed the relative orientation of the C-termini within the dimeric protein from antiparallel to parallel, resulting in only transient osmotic activation. These results suggest that ProP is in monomer-dimer equilibrium in vivo. Dimerization may be mediated by C-terminal coiled-coil formation and/or by interactions between other structural domains, which in turn facilitate C terminal coiled-coil formation. Antiparallel coiled-coil formation is required for activation of ProP at low osmolality. PMID- 16042395 TI - Biphasic kinetics of the colchicine-tubulin interaction: role of amino acids surrounding the A ring of bound colchicine molecule. AB - Isotypes of vertebrate tubulin have variable amino acid sequences, which are clustered at their C-terminal ends. Isotypes bind colchicine at different on rates and affinity constants. The kinetics of colchicine binding to purified (unfractionated) brain tubulin have been reported to be biphasic under pseudo first-order conditions. Experiments with individual isotypes established that the presence of beta(III) in the purified tubulin is responsible for the biphasic kinetics. Because the isotypes mainly differ at the C termini, the colchicine binding kinetics of unfractionated tubulin and the beta(III) isotype, cleaved at the C termini, have been tested under pseudo-first-order conditions. Removal of the C termini made no difference to the nature of the kinetics. Sequence alignment of different beta isotypes of tubulin showed that besides the C terminal region, there are differences in the main body as well. To establish whether these differences lie at the colchicine-binding site or not, homology modeling of all beta-tubulin isotypes was done. We found that the isotypes differed from each other in the amino acids located near the A ring of colchicine at the colchicine-binding site on beta tubulin. While the beta(III) isotype has two hydrophilic residues (serine(242) and threonine(317)), both beta(II) and beta(IV) have two hydrophobic residues (leucine(242) and alanine(317)). beta(II) has isoleucine at position 318, while beta(III) and beta(IV) have valine at that position. Thus, these alterations in the nature of the amino acids surrounding the colchicine site could be responsible for the different colchicine-binding kinetics of the different isotypes of tubulin. PMID- 16042396 TI - Translocation of beta-galactosidase mediated by the cell-penetrating peptide pep 1 into lipid vesicles and human HeLa cells is driven by membrane electrostatic potential. AB - The cell-penetrating peptide (CPP) pep-1 is capable of introducing large proteins into different cell lines, maintaining their biological activity. Two possible mechanisms have been proposed to explain the entrance of other CPPs in cells, endosomal-dependent and independent types. In this work, we evaluated the molecular mechanisms of pep-1-mediated cellular uptake of beta-galactosidase (beta-Gal) from Escherichia coli in large unilamellar vesicles (LUV) and HeLa cells. Fluorescence spectroscopy was used to evaluate the translocation process in model systems (LUV). Immunofluorescence microscopy was used to study the translocation in HeLa cells. Enzymatic activity detection enabled us to monitor the internalization of beta-Gal into LUV and the functionality of the protein in the interior of HeLa cells. Beta-Gal translocated into LUV in a transmembrane potential-dependent manner. Likewise, the extent of beta-Gal incorporation was extensively decreased in depolarized cells. Furthermore, beta-Gal uptake efficiency and kinetics were temperature-independent, and beta-Gal did not colocalize with endosomes, lysosomes, or caveosomes. Therefore, beta-Gal translocation was not associated with the endosomal pathway. Although an excess of pep-1 was mandatory for beta-Gal translocation in vivo, transmembrane pores were not formed as concluded from the trypan blue exclusion method. These results altogether indicated that protein uptake both in vitro with LUV and in vivo with HeLa cells was mainly, if not solely, dependent on negative transmembrane potential across the bilayer, which suggests a physical mechanism governed by electrostatic interactions between pep-1 (positively charged) and membranes (negatively charged). PMID- 16042397 TI - Analysis of a novel diacylglycerol kinase from Dictyostelium discoideum: DGKA. AB - Diacylglycerol kinases (DGKs) catalyze the ATP-dependent phosphorylation of diacylglycerols to generate phosphatidic acid and have been investigated in prokaryotic and eukaryotic organisms. Recently, a protein that is significantly similar to human DGK-theta, DGKA, was identified in Dictyostelium discoideum. It has been shown to possess DGK activity when assayed using a medium-chain diacylglycerol, 1,2-dioctanoyl-sn-glycerol (DiC8). A complete understanding of DGK catalytic and regulatory mechanisms, as well as physiological roles, requires an understanding of its biochemical and kinetic properties. This report presents an analysis of these properties for DGKA. The enzyme catalyzes the phosphorylation of DiC8, and another medium-chain DAG, DiC6 (1,2-dihexanoyl-sn glycerol), in a Michaelis-Menten manner. Interestingly, the kinetics of DGKA using physiologically relevant long-chain DAGs was dependent on substrate surface concentration and the detergent that was used. DGKA displayed Michaelis-Menten kinetics with respect to bulk substrate concentration (1,2-dioleoyl-sn-glycerol) in octyl glucoside mixed micelles when the surface substrate concentration was at or below 3.5 mol %. At higher surface concentrations, however, there was a sigmoidal relationship between the initial velocity and bulk substrate concentration. In contrast, DGKA displayed sigmoidal kinetics with respect to bulk substrate concentrations at all surface concentrations in Triton X-100 mixed micelles. Finally, we show the catalytic activity of DGKA was significantly enhanced by phosphatidylserine (PS) and phosphatidic acid (PA). PMID- 16042398 TI - Solid-state nuclear magnetic resonance relaxation studies of the interaction mechanism of antimicrobial peptides with phospholipid bilayer membranes. AB - An 18-residue peptide, KWGAKIKIGAKIKIGAKI-NH(2) was designed to form amphiphilic beta-sheet structures when bound to lipid bilayers. The peptide possesses high antimicrobial activity when compared to naturally occurring linear antimicrobial peptides, most of which adopt an amphipathic alpha-helical conformation upon binding to the lipids. The perturbation of the bilayer by the peptide was studied by static (31)P and (2)H solid-state NMR spectroscopy using POPC and POPG/POPC (3/1) bilayer membranes with sn-1 chain perdeuterated POPC and POPG as the isotopic labels. (31)P NMR powder spectra exhibited two components for POPG/POPC bilayers upon addition of the peptide but only a slight change in the line shape for POPC bilayers, indicating that the peptide selectively disrupted the membrane structure consisting of POPG lipids. (2)H NMR powder spectra indicated a reduction in the lipid chain order for POPC bilayers and no significant change in the ordering for POPG/POPC bilayers upon association of the peptide with the bilayers, suggesting that the peptide acts as a surface peptide in POPG/POPC bilayers. Relaxation rates are more sensitive to the motions of the membranes over a large range of time scales. Longer (31)P longitudinal relaxation times for both POPG and POPC in the presence of the peptide indicated a direct interaction between the peptide and the POPG/POPC bilayer membranes. (31)P longitudinal relaxation studies also suggested that the peptide prefers to interact with the POPG phospholipids. However, inversion-recovery (2)H NMR spectroscopic experiments demonstrated a change in the relaxation rate of the lipid acyl chains for both the POPC membranes and the POPG/POPC membranes upon interaction with the peptide. Transverse relaxation studies indicated an increase in the spectral density of the collective membrane motion caused by the interaction between the peptide and the POPG/POPC membrane. The experimental results demonstrate significant dynamic changes in the membrane in the presence of the antimicrobial peptide and support a carpet mechanism for the disruption of the membranes by the antimicrobial peptide. PMID- 16042399 TI - Electrostatic effects on the stability of discoidal high-density lipoproteins. AB - High-density lipoproteins (HDL) remove cholesterol from peripheral tissues and thereby help to prevent atherosclerosis. Nascent HDL are discoidal complexes composed of a phospholipid bilayer surrounded by protein alpha-helices that are thought to form extensive stabilizing interhelical salt bridges. Earlier we showed that HDL stability, which is necessary for HDL functions, is modulated by kinetic barriers. Here we test the role of electrostatic interactions in the kinetic stability by analyzing the effects of salt, pH, and point mutations on model discoidal HDL reconstituted from human apolipoprotein C-1 (apoC-1) and dimyristoyl phosphatidylcholine (DMPC). Circular dichroism, Trp fluorescence, and light scattering data show that molar concentrations of NaCl or Na(2)SO(4) increase the apparent melting temperature of apoC-1:DMPC complexes by up to 20 degrees C and decelerate protein unfolding. Arrhenius analysis shows that 1 M NaCl stabilizes the disks by deltaDeltaG* approximately equal 3.5 kcal/mol at 37 degrees C and increases the activation energy of their denaturation and fusion by deltaE(a) approximately equal deltaDeltaH* approximately equal 13 kcal/mol, indicating that the salt-induced stabilization is enthalpy-driven. Denaturation studies in various solvent conditions (pH 5.7-8.2, 0-40% sucrose, 0-2 M trimethylamine N-oxide) suggest that the salt-induced disk stabilization results from ionic screening of unfavorable short-range Coulombic interactions. Thus, the dominant electrostatic interactions in apoC-1:DMPC disks are destabilizing. Comparison of the salt effects on the protein:lipid complexes of various composition reveals an inverse correlation between the lipoprotein stability and the salt-induced stabilization and suggests that short-range electrostatic interactions significantly contribute to lipoprotein stability: the better optimized these interactions are, the more stable the complex is. PMID- 16042400 TI - Cyanine dye N744 inhibits tau fibrillization by blocking filament extension: implications for the treatment of tauopathic neurodegenerative diseases. AB - Tau fibrillization is a potential therapeutic target for Alzheimer's and other neurodegenerative diseases. Small molecules capable of both inhibiting aggregation and promoting filament disaggregation have been discovered, but knowledge of their mechanism of action and potential for testing in biological models is fragmentary. To clarify these issues, the interaction between a small molecule inhibitor of tau fibrillization, 3,3'-bis(beta-hydroxyethyl)-9-ethyl 5,5'-dimethoxythiacarbocyanine iodide (N744), and full-length four-repeat tau protein was characterized in vitro using transmission electron microscopy and fluorescence spectroscopy. Analysis of reaction time courses performed in the presence of anionic fibrillization inducers revealed that increasing concentrations of N744 decreased the total filament length without modulating lag time, indicating that filament extension but not nucleation was affected by inhibitor under the conditions that were investigated. Critical concentration measurements confirmed that N744 shifted equilibria at filament ends away from the fibrillized state, resulting in endwise filament disaggregation when it was added to synthetic filaments. Both increasing bulk tau concentrations and filament stabilizing modifications such as pseudophosphorylation and glycation antagonized N744 activity. The results illustrate the importance of mechanism for the design and interpretation of pharmacological studies in biological models of tau aggregation. PMID- 16042401 TI - Thermodynamics of nucleotide binding to actomyosin V and VI: a positive heat capacity change accompanies strong ADP binding. AB - We have measured the energetics of ATP and ADP binding to single-headed actomyosin V and VI from the temperature dependence of the rate and equilibrium binding constants. Nucleotide binding to actomyosin V and VI can be modeled as two-step binding mechanisms involving the formation of collision complexes followed by isomerization to states with high nucleotide affinity. Formation of the actomyosin VI-ATP collision complex is much weaker and slower than for actomyosin V. A three-step binding mechanism where actomyosin VI isomerizes between two conformations, one competent to bind ATP and one not, followed by rapid ATP binding best accounts for the data. ADP binds to actomyosin V more tightly than actomyosin VI. At 25 degrees C, the strong ADP-binding equilibria are comparable for actomyosin V and VI, and the different overall ADP affinities arise from differences in the ADP collision complex affinity. The actomyosin-ADP isomerization leading to strong ADP binding is entropy driven at >15 degrees C and occurs with a large, positive change in heat capacity (DeltaC(P) degrees ) for both actomyosin V and VI. Sucrose slows ADP binding and dissociation from actomyosin V and VI but not the overall equilibrium constants for strong ADP binding, indicating that solvent viscosity dampens ADP-dependent kinetic transitions, presumably a tail swing that occurs with ADP binding and release. We favor a mechanism where strong ADP binding increases the dynamics and flexibility of the actomyosin complex. The heat capacity (DeltaC(P) degrees ) and entropy (DeltaS degrees ) changes are greater for actomyosin VI than actomyosin V, suggesting different extents of ADP-induced structural rearrangement. PMID- 16042402 TI - ATP hydrolysis promotes interactions between the extracellular ends of transmembrane segments 1 and 11 of human multidrug resistance P-glycoprotein. AB - P-glycoprotein (P-gp, ABCB1) actively pumps a broad range of structurally unrelated cytotoxic compounds out of the cell. It has two homologous halves that are joined by a linker region. Each half has a transmembrane (TM) domain containing six TM segments and a nucleotide-binding domain (NBD). Cross-linking studies have shown that the drug-binding pocket is at the interface between the TM domains. The two NBDs interact to form the ATP-binding sites. Coupling of ATP hydrolysis to drug efflux has been postulated to occur by conversion of the binding pocket from a high-affinity to a low-affinity state through alterations in the packing of the TM segments. TM 11 has also been reported to be important for drug binding. Here, we used cysteine-scanning mutagenesis and oxidative cross linking to test for changes in the packing of TM 11 during ATP hydrolysis. We generated 350 double cysteine mutants that contained one cysteine at the extracellular end of TM11 and another cysteine at the extracellular ends of TMs 1, 3, 4, 5, or 6. The mutants were expressed in HEK293 cells and treated with oxidant in the absence or presence of ATP. Cross-linked product was not detected in SDS-PAGE gels in the absence of ATP. By contrast, cross-linked product was detected in mutants M68C(TM1)/Y950C(TM11), M68C(TM1)/Y953C(TM11), M68C(TM1)/A954C(TM11), M69C(TM1)/A954C(TM11), and M69C(TM1)/ F957C(TM11) in the presence of ATP but not with ADP or AMP.PNP. These results indicate that rearrangement of TM11 may contribute to the release of drug substrate during ATP hydrolysis. PMID- 16042403 TI - Aggregation and porin-like channel activity of a beta sheet peptide. AB - Beta sheet peptides (e.g., amyloid beta) are known to form ion channels in lipid bilayers possibly through aggregation, though the channel structure is not clear. We have recently reported that a short beta sheet peptide, (xSxG)(6), forms porin like voltage-gated channels in lipid bilayers [Thundimadathil et al. (2005) Biochem. Biophys. Res. Commun. 330, 585-590]. To account for the porin-like activity, oligomerization of the peptide into a beta barrel-like structure was proposed. In this work, peptide aggregation in aqueous and membrane environments and a detailed study of channel properties were performed to gain insight into the mechanism of channel formation. The complex nature of the channel was revealed by kinetic analysis and the occurrence of interconverting multiple conductance states. Ion channels were inhibited by Congo red, suggesting that the peptide aggregates are the active channel species. Peptide aggregation and fibril formation in water were confirmed by electron microscopy (EM) and Congo red binding studies. Furthermore, oligomeric structures in association with lipid bilayers were detected. Circular dichroism of peptide-incorporated liposomes and peptide-lipid binding studies using EM suggest a lipid-induced beta sheet aggregation. Gel electrophoresis of peptide-incorporated liposomes showed dimeric and multimeric structures. Taken together, this work indicates insertion of (xSxG)(6) as oligomers into the lipid bilayer, followed by rearrangement into a beta barrel-like pore structure. A large peptide pore comprising several individual beta sheets or smaller beta sheet aggregates is expected to have a complex behavior in membranes. A dyad repeat sequence and the presence of glycine, serine, and hydrophobic residues in a repeated pattern in this peptide may be providing a favorable condition for the formation of a beta barrel-like structure in lipid bilayers. PMID- 16042404 TI - Coacervation is promoted by molecular interactions between the PF2 segment of fibrillin-1 and the domain 4 region of tropoelastin. AB - In forming elastic fibers, microfibrils act as the scaffold sites for depositing the elastin precursor tropoelastin. We examined key binding interactions that promote massive tropoelastin association through coacervation. Using a segment of the microfibril protein fibrillin-1, PF2, known to bind full-length tropoelastin, we mapped its interaction site to the N-terminal region of tropoelastin bounded by domains 2 and 18. Precise contact residues between domain 4 of tropoelastin and domain 16 of fibrillin-1 were discovered through a novel combination of transglutaminase cross-linking and mass spectroscopy, with contact sites at residues K38 of tropoelastin and Q669 of fibrillin-1. This is the first report of a role for this region of tropoelastin in microfibril interactions. The addition of PF2 thermodynamically facilitated the coacervation of tropoelastin, resulting in smaller changes in entropy and enthalpy values for the coacervating system. A novel multicomponent in vitro tropoelastin assembly reaction system demonstrated that amassed tropoelastin was spatially and preferentially directed to surfaces coated with PF2 as expected for organized three-dimensional distribution during tissue elastogenesis. This study underscores the role of this part of fibrillin-1 as an anchor point for tropoelastin at the microfibril-elastin junction during the initial stages of elastic fiber assembly. PMID- 16042405 TI - Acyl-coenzyme A binding protein expression alters liver fatty acyl-coenzyme A metabolism. AB - Although studies in vitro and in yeast suggest that acyl-CoA binding protein ACBP may modulate long-chain fatty acyl-CoA (LCFA-CoA) distribution, its physiological function in mammals is unresolved. To address this issue, the effect of ACBP on liver LCFA-CoA pool size, acyl chain composition, distribution, and transacylation into more complex lipids was examined in transgenic mice expressing a higher level of ACBP. While ACBP transgenic mice did not exhibit altered body or liver weight, liver LCFA-CoA pool size increased by 69%, preferentially in saturated and polyunsaturated, but not monounsaturated, LCFA CoAs. Intracellular LCFA-CoA distribution was also altered such that the ratio of LCFA-CoA content in (membranes, organelles)/cytosol increased 2.7-fold, especially in microsomes but not mitochondria. The increased distribution of specific LCFA-CoAs to the membrane/organelle and microsomal fractions followed the same order as the relative LCFA-CoA binding affinity exhibited by murine recombinant ACBP: saturated > monounsaturated > polyunsaturated C14-C22 LCFA CoAs. Consistent with the altered microsomal LCFA-CoA level and distribution, enzymatic activity of liver microsomal glycerol-3-phosphate acyltransferase (GPAT) increased 4-fold, liver mass of phospholipid and triacylglyceride increased nearly 2-fold, and relative content of monounsaturated C18:1 fatty acid increased 44% in liver phospholipids. These effects were not due to the ACBP transgene altering the protein levels of liver microsomal acyltransferase enzymes such as GPAT, lysophosphatidic acid acyltransferase (LAT), or acyl-CoA cholesterol acyltransferase 2 (ACAT-2). Thus, these data show for the first time in a physiological context that ACBP expression may play a role in LCFA-CoA metabolism. PMID- 16042406 TI - A Glu113Ala mutation within a factor VIII Ca2+-binding site enhances cofactor interactions in factor Xase. AB - We recently identified an acidic-rich segment in the A1 domain of factor VIII (residues 110-126) that functions in the coordination of Ca(2+), an ion necessary for cofactor activity [Wakabayashi et al. (2004) J. Biol. Chem. 279, 12677 12684]. Mutagenesis studies showed that replacement of residue Glu113 with Ala (E113A) yielded a factor VIII point mutant possessing increased specific activity as determined by a one-stage clotting assay. Mutagenesis at this site suggested that substitution with relatively small, nonpolar residues was well tolerated, whereas replacement with a number of polar or charged residues appeared detrimental to activity. Ala substitution resulted in the greatest enhancement, yielding an approximately 2-fold increased specific activity. Time course experiments following reaction with thrombin revealed similar rates of activation and inactivation of E113A as observed for the wild type. Results from factor Xa generation assays showed minimal differences in kinetic parameters and factor IXa affinity for E113A and wild-type factor VIIIa when run in the presence of synthetic phospholipid vesicles, whereas factor VIIIa E113A displayed an approximately 4-fold greater affinity for factor IXa compared with factor VIIIa wild type in reactions run on the platelet membrane surface. This latter effect may be attributed, in part, to a 2-fold increased affinity of factor VIIIa E113A for the platelet membrane. Considering that low levels of factors VIIIa and IXa are generated during clotting in plasma, the increased cofactor specific activity observed for E113A factor VIII may result from its enhanced affinity for factor IXa on the physiological membrane. PMID- 16042407 TI - Antifreeze proteins of the beetle Dendroides canadensis enhance one another's activities. AB - Larvae of the beetle Dendroides canadensis produce a family of 13 antifreeze proteins (DAFPs), four of which are in the hemolymph. Antifreeze proteins lower the noncolligative freezing point of water (in the presence of ice) below the melting point, producing a difference between the freezing and melting points termed thermal hysteresis. This activity (THA) is dependent upon DAFP specific activity, concentration, and the presence of enhancers. Enhancers may be low molecular mass enhancers, such as glycerol, or other proteins. The protein enhancers complex with the DAFPs, thereby blocking a larger surface area of the potential seed ice crystal and consequently lowering the freezing point. A yeast two-hybrid screen was performed using certain hemolymph DAFPs as "bait" in an effort to identify endogenous protein enhancers. Among the positive proteins identified as interacting with the bait DAFPs, and confirmed by co immunoprecipitation, were other DAFPs. When pure DAFPs were added to one another, those identified by the yeast two-hybrid screen as interacting with one another exhibited a synergistic enhancement of thermal hysteresis activity. In contrast, those DAFPs which the screen indicated did not interact failed to enhance one anothers' activities. DAFPs-1 and -2 interact and enhance one another. Point mutations of one of the interacting DAFPs (DAFP-2) indicated that both of the two amino acid residues that differ between DAFPs-1 and -2 were required for interaction. Glycerol enhanced the THA of the DAFPs only when DAFPs known to interact were present in the test solution. Addition of glycerol to a test solution containing only one DAFP did not produce enhancement. Therefore, glycerol enhances activity by stimulating interactions between DAFPs. PMID- 16042408 TI - Regulation of peroxisome proliferator-activated receptor alpha by protein kinase C. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) is a nuclear receptor activated by fatty acids, hypolipidemic drugs, and peroxisome proliferators (PPs). Like other nuclear receptors, PPARalpha is a phosphoprotein whose activity is affected by a variety of growth factor signaling cascades. In this study, the effects of protein kinase C (PKC) on PPARalpha activity were explored. In vivo phosphorylation studies in COS-1 cells transfected with murine PPARalpha showed that the level of phosphorylated PPARalpha is increased by treatment with the PP Wy-14,643 as well as the PKC activator phorbol myristol acetate (PMA). In addition, inhibitors of PKC decreased Wy-14,643-induced PPARalpha activity in a variety of reporter assays. Overexpressing PKCalpha, beta, -delta, and -zeta affected both basal and Wy-14,643-induced PPARalpha activity. Four consensus PKC phosphorylation sites are contained within the DNA binding (C-domain) and hinge (D-domain) regions of rat PPARalpha (S110, T129, S142, and S179), and their contribution to receptor function was examined. Mutation of T129 or S179 to alanine prevented heterodimerization of PPARalpha with RXRalpha, lowered the level of phosphorylation by PKCalpha and PKCdelta in vitro, and lowered the level of phosphorylation of transfected PPARalpha in transfected cells. In addition, the T129A mutation prevented PPARalpha from binding DNA in an electromobility shift assay. Together, these studies demonstrate a direct role for PKC in the regulation of PPARalpha, and suggest several PKCs can regulate PPARalpha activity through multiple phosphorylation sites. PMID- 16042409 TI - Heme reduction by intramolecular electron transfer in cysteine mutant myoglobin under carbon monoxide atmosphere. AB - Human myoglobin (Mb) possesses a unique cysteine (Cys110), whereas other mammalian Mbs do not. To investigate the effect of a cysteine residue on Mb, we introduced cysteine to various sites on the surface of sperm whale Mb (K56C, V66C, K96C, K102C, A125C, and A144C) by mutation. The cysteines were inserted near the end of alpha-helices, except for V66C, where the cysteine was introduced in the middle of an alpha-helix. Reduction of the heme was observed for each mutant metMb by incubation at 37 degrees C under carbon monoxide atmosphere, which was much faster than reduction of wild-type metMb under the same condition. Heme reduction did not occur significantly under nitrogen or oxygen atmospheres. The rate constant for heme reduction increased for higher mutant Mb concentration, whereas it did not change significantly when the CO concentration was reduced from 100% CO to 50% CO with 50% O(2). The similarity in the rate constants with different CO concentrations indicates that CO stabilizes the reduced heme by coordination to the heme iron. SDS-PAGE analysis showed that mutant Mb dimers were formed by incubation under CO atmosphere but not under air. These dimers were converted back to Mb monomers by an addition of 2 mercaptoethanol, which showed formation of a Mb dimer through a disulfide bond. The rate constant decreased in general as the heme-cysteine distance was increased, although V66C Mb exhibited a very small rate constant. Since V66 is placed in the middle of an alpha-helix, steric hindrance would occur and prevent formation of a dimer when the cysteine residues of two different V66C Mb molecules interact with each other. The rate constants also decreased for K56C and A144C Mbs presumably because of the electrostatic repulsion during dimer formation, since they are relatively charged around the inserted cysteine. PMID- 16042410 TI - Allosteric models for multimeric proteins: oxygen-linked effector binding in hemocyanin. AB - In many crustaceans, changing concentrations of several low molecular weight compounds modulates hemocyanin oxygen binding, resulting in lower or higher oxygen affinities of the pigment. The nonphysiological effector caffeine and the physiological modulator urate, the latter accumulating in the hemolymph of the lobster Homarus vulgaris during hypoxia, increase hemocyanin oxygen affinity and decrease cooperativity of oxygen binding. To derive a model that describes the mechanism of allosteric interaction between hemocyanin and oxygen in the presence of urate or caffeine, studies of oxygen, urate, and caffeine binding to hemocyanin were performed. Exposure of lobster hemocyanin to various pH values between 7.25 and 8.15 resulted in a decrease of p50. In this pH interval, p50 decreases from 95 to 11 Torr without effectors and from 49 to 6 Torr and from 34 to 5 Torr in the presence of 1 mM urate or caffeine, respectively. Thus, the allosteric effects induced by protons and urate or caffeine are coupled. In contrast, isothermal titration calorimetry did not reveal any differences in binding enthalpy (DeltaH degrees ) for urate or caffeine under either normoxic or hypoxic conditions at different pH values. Despite these apparently conflicting results, they can be explained by the nested MWC model if two different types of modulator binding sites are assumed, an allosteric and a nonallosteric type of site. Simulations of in vivo conditions with this model indicate that the naturally occurring modulator urate is physiologically relevant in H. vulgaris only during hypoxic conditions, i.e., either during environmental oxygen limitation or extensive exercise. PMID- 16042411 TI - Characterization of an Escherichia coli sulfite oxidase homologue reveals the role of a conserved active site cysteine in assembly and function. AB - We report the biochemical and biophysical characterization of YedYZ, a sulfite oxidase homologue from Escherichia coli. YedY is a soluble catalytic subunit with a twin arginine leader sequence for export to the periplasm by the Tat translocation system. YedY is the only molybdoenzyme so far isolated from E. coli with the Mo-MPT form of the molybdenum cofactor. The electron paramagnetic resonance (EPR) signal of the YedY molybdenum is similar to that of known Mo-MPT containing enzymes, with the exception that only the Mo(IV) --> Mo(V) transition is observed, with a midpoint potential of 132 mV. YedZ is a membrane-intrinsic cytochrome b with six putative transmembrane helices. The single heme b of YedZ has a midpoint potential of -8 mV, determined by EPR spectroscopy of YedZ enriched membrane preparations. YedY does not associate strongly with YedZ on the cytoplasmic membrane. However, mutation of the YedY active site Cys102 to Ser results in very efficient targeting of YedY to YedZ in the membrane, demonstrating a clear role for YedZ as the membrane anchor for YedY. Together, YedYZ comprise a well-conserved bacterial heme-molybdoenzyme found in a variety of bacteria that can be assigned to the sulfite oxidase class of enzyme. PMID- 16042412 TI - Papain-like protease 2 (PLP2) from severe acute respiratory syndrome coronavirus (SARS-CoV): expression, purification, characterization, and inhibition. AB - Viral proteases are essential for pathogenesis and virulence of severe acute respiratory syndrome coronavirus (SARS-CoV). Little information is available on SARS-CoV papain-like protease 2 (PLP2), and development of inhibitors against PLP2 is attractive for antiviral therapy. Here, we report the characterization of SARS-CoV PLP2 (from residues 1414 to 1858) purified from baculovirus-infected insect cells. We demonstrate that SARS-CoV PLP2 by itself differentially cleaves between the amino acids Gly180 and Ala181, Gly818 and Ala819, and Gly2740 and Lys2741 of the viral polypeptide pp1a, as determined by reversed-phase high performance liquid chromatography analysis coupled with mass spectrometry. This protease is especially selective for the P1, P4, and P6 sites of the substrate. The study demonstrates, for the first time among coronaviral PLPs, that the reaction mechanism of SARS-CoV PLP2 is characteristic of papain and compatible with the involvement of the catalytic dyad (Cys)-S(-)/(His)-Im(+)H ion pair. With a fluorogenic inhibitor-screening platform, we show that zinc ion and its conjugates potently inhibit the enzymatic activity of SARS-CoV PLP2. In addition, we provided evidence for evolutionary reclassification of SARS-CoV. The results provide important insights into the biochemical properties of the coronaviral PLP family and a promising therapeutic way to fight SARS-CoV. PMID- 16042413 TI - A mechanistic investigation of the thiol-disulfide exchange step in the reductive dehalogenation catalyzed by tetrachlorohydroquinone dehalogenase. AB - Tetrachlorohydroquinone dehalogenase catalyzes the reductive dehalogenation of tetrachloro- and trichlorohydroquinone to give 2,6-dichlorohydroquinone in the pathway for degradation of pentachlorophenol by Sphingobium chlorophenolicum. Previous work has suggested that this enzyme may have originated from a glutathione-dependent double bond isomerase such as maleylacetoacetate isomerase or maleylpyruvate isomerase. While some of the elementary steps in these two reactions may be similar, the final step in the dehalogenation reaction, a thiol disulfide exchange reaction that removes glutathione covalently bound to Cys13, certainly has no counterpart in the isomerization reaction. The thiol-disulfide exchange reaction does not appear to have been evolutionarily optimized. There is little specificity for the thiol; many thiols react at high rates. TCHQ dehalogenase binds the glutathione involved in the thiol-disulfide exchange reaction very poorly and does not alter its pK(a) in order to improve its nucleophilicity. Remarkably, single-turnover kinetic studies show that the enzyme catalyzes this step by approximately 10000-fold. This high reactivity requires an as yet unidentified protonated group in the active site. PMID- 16042414 TI - Atypical recognition of particular DNA sequences by the archaeal chromosomal MC1 protein. AB - The MC1 protein is a chromosomal protein likely involved in the DNA compaction of some methanogenic archaea. This small and monomeric protein, structurally unrelated to other DNA binding proteins, bends DNA sharply. By studying the protein binding to various kinds of kinked DNA, we have previously shown that MC1 is able to discriminate between different deformations of the DNA helix. Here we investigate its capacity to recognize particular DNA sequences by using a SELEX procedure. We find that MC1 is able to preferentially bind to a 15 base pair motif [AAAAACACAC(A/C)CCCC]. The structural parameters of this sequence are characterized by molecular dynamics simulation experiments, and the binding mode of the protein to the DNA is studied by footprinting experiments. Our results strongly suggest that the protein realizes an indirect readout of the DNA sequence by binding to the DNA minor groove. PMID- 16042415 TI - Effect of DNA polymerase beta loop variants on discrimination of O6 methyldeoxyguanosine modification present in the nucleotide versus template substrate. AB - We have examined the mechanism of DNA polymerase beta (pol beta) lesion discrimination using alkylated dNTP versus alkylated DNA template substrates and the pol beta variants R253M and E249K. Both of these amino acid variants are located in the loop region of the palm domain and are known to play a role in pol beta fidelity and discrimination of 3'-azido-3'-deoxythymidine triphosphate substrates. We observed that these variants affect O(6)-methyldeoxyguanosine- (m6G-) modified dNTP discrimination without affecting m6G template translesion synthesis. Under steady-state conditions, the ratio of inherent reactivity values for the m6dGTP substrate relative to the dGTP substrate was greater for both variant polymerases than for wild-type (WT) pol beta. Biochemical assays of translesion synthesis using m6G lesion-containing templates demonstrated no significant differences between the variants and WT. Using N-methyl-N-nitrosourea (MNU-) modified DNA templates in the HSV-tk in vitro assay, no difference among the enzymes in the frequency of alkylation-induced G to A transition mutations was observed. However, differences among the polymerases in the frequency of alkylation-induced C to A transversions were observed, consistent with a mutator tendency for E249K and an antimutator tendency for R253M. We conclude that a specific interaction at the loop of the palm domain is involved in pol beta discrimination of the m6G lesion when present on the incoming dNTP substrate but not when present in the DNA template. Our data support a role for the flexible loop in pol beta error discrimination. PMID- 16042416 TI - Combinatorial selection, inhibition, and antiviral activity of DNA thioaptamers targeting the RNase H domain of HIV-1 reverse transcriptase. AB - Despite the key role played by the RNase H of human immunodeficiency virus-1 reverse transcriptase (HIV-1 RT) in viral proliferation, only a few inhibitors of RNase H have been reported. Using in vitro combinatorial selection methods and the RNase H domain of the HIV RT, we have selected double-stranded DNA thioaptamers (aptamers with selected thiophosphate backbone substitutions) that inhibit RNase H activity and viral replication. The selected thioaptamer sequences had a very high proportion of G residues. The consensus sequence for the selected thioaptamers showed G clusters separated by single residues at the 5'-end of the sequence. Gel electrophoresis mobility shift assays and nuclear magnetic resonance spectroscopy showed that the selected thioaptamer binds to the isolated RNase H domain, but did not bind to a structurally similar RNase H from Escherichia coli. The lead thioaptamer, R12-2, showed specific binding to HIV-1 RT with a binding constant (K(d)) of 70 nM. The thioaptamer inhibited the RNase H activity of intact HIV-1 RT. In cell culture, transfection of thioaptamer R12-2 (0.5 microg/mL) markedly inhibited viral production and exhibited a dose response of inhibition with R12-2 concentrations ranging from 0.03 to 2.0 microg/mL (IC(50) < 100 nM). Inhibition was also seen across a wide range of virus inoculum, ranging from a multiplicity of infection (moi) of 0.0005 to 0.05, with a reduction of the level of virus production by more than 50% at high moi. Suppression of virus was comparable to that seen with AZT when moi or = CAC), where the cloudy solution becomes clear. For cross linked HASE latex, anionic surfactant binds only noncooperatively to the latex and causes it to swell. For cationic surfactant, electrostatic interaction occurs at very low surfactant concentrations, resulting in phase separation. With further increase in surfactant concentration, noncooperative hydrophobic and cooperative hydrophobic interactions dominate the binding at low and high surfactant concentrations, respectively. For anionic and cationic surfactant systems, the CAC is lower than the critical micelle concentration (CMC) of surfactants in water. In addition, counterion condensation plays an important role during the binding interaction between HASE latex and ionic surfactants. In the case of nonionic surfactants, free surfactant micelles are formed in solution due to their relatively low CMC values, and HASE latexes are directly solubilized into the micellar core of nonionic surfactants. PMID- 16042435 TI - Self-organization of the ternary didecyldimethylammonium bromide/octyl-beta-D glucopyranoside/water system. AB - The spontaneous and thermodynamically stable mixed vesicles constituted by a double-chain cationic surfactant with 10 carbon atoms hydrophobic tail, didecyldimethylammonium bromide (di-C(10)DMAB), and a nonionic single-chain surfactant, octyl-beta-d-glucopyranoside (OBG), have been characterized in aqueous media by means of a series of experimental techniques, as well as a theoretical approach. Conductivity data allow for the determination of the concentrations at which the monomer-to-vesicle (CVC) and/or vesicle-to-micelle (CMC) transitions occur. Electrophoretic mobilities, obtained from laser-doppler electrophoresis experiments, permit the determination of zeta-potentials and, from them, the surface charge density of the vesicle aggregates. Cryogenic transmission electron microscopy (cryo-TEM) provides pictures of the vesicles, their size and shape being, thus, determined. Finally, the sensitivity of the emission spectra of some fluorescent probes, such as the cationic TNS and the nonionic PRODAN, to the polarity of the environment, allow for a complete study of different pre- and post-vesicle microdomains, of variable rigidity and micropolarity. This, in turn, yield interesting information about the vesicle surface and bilayer, as well as, about the existence of clusters and/or nanoaggregates prior to the formation of vesicles, as was proposed by us in a previous paper. PMID- 16042436 TI - Specific interactions of poly(4-vinyl phenol) gel with cationic and anionic surfactants. AB - Binding behaviors of ionic surfactants (decyl- and dodecyltrimethylammonium bromide (C(10)TAB, C(12)TAB), sodium decane sulfonate (SDeSo), and sodium dodecyl sulfate (SDS)) to poly(4-vinyl phenol) (P4VPh) gel were investigated to elucidate a specific swelling behavior that has been found for P4VPh gel in aqueous solutions of tetraalkylammonium salts. With increasing cationic surfactant concentration, P4VPh gel significantly deswelled and then remarkably reswelled at a concentration somewhat below the respective cmc values. On the other hand, in the case of the anionic surfactants, the gel only showed a marked swelling at a concentration just below the respective cmc values. A similar charge-specific behavior of the surfactants was also found for the P4VPh dispersion system studied with a UV-vis spectroscopy; namely, in the cationic surfactant-P4VPh systems, the turbidity of the dispersion first increased with increasing the surfactant concentration and then decreased. This result suggests that aggregation of P4VPh particles first occurred and finally the particles were solubilized. A red shift followed by a blue shift observed for a pi-pi absorption of phenol at around 278 nm was also consistent with the aggregation solubilization behavior. In the anionic surfactant-P4VPh system, however, only solubilization of the polymer particle was observed, and the UV peak only showed a blue shift. All these results in the gel and the dispersion systems strongly suggest that the cation-pi interaction is involved in the binding of the cationic surfactants to P4VPh. PMID- 16042437 TI - Micellar solutions of sulfobetaine surfactants in water-ethylene glycol mixtures: surface tension, fluorescence, spectroscopic, conductometric, and kinetic studies. AB - Micellization in water-ethylene glycol (EG) N-dodecyl, N-tetradecyl, and N hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (SB3-12, SB3-14, and SB3-16, respectively) micellar solutions, with the weight percent of EG changing within the range 0-40, was studied by means of surface tension measurements. Information about the influence of the added EG on the aggregation number of the sulfobetaine micelles and on the polarity of the interfacial region of micelles was obtained through fluorescence and spectroscopic measurements. Surface tension measurements also provide information about the dependence of the surface excess concentration, the minimum area per surfactant molecule, the surface pressure at the cmc, and the standard Gibbs energy of adsorption on the added weight percent of the organic solvent. The Gordon parameter of the water-EG mixtures was also estimated by means of surface tension measurements. The thermodynamic and structural changes originated by the presence of EG control the micellar kinetic effects observed in the reaction methyl 4-nitrobenzenesulfonate + Br(-) occurring in the water-EG sulfobetaine micellar solutions. Information about the distribution of bromide ions between the bulk and micellar pseudophases was obtained through conductivity measurements. The kinetic micellar effects were quantitatively explained by using the pseudophase kinetic model. PMID- 16042438 TI - Synthesis and properties of cationic oligomeric surfactants. AB - Three series of new oligomeric cationic surfactants were synthesized. These amphiphiles are trimeric and tetrameric oligomeric quaternary ammonium chlorides, with spacer groups of different lengths separating the individual surfactant fragments. The properties of the compounds, such as Krafft temperatures, surface activity, micellization, viscosifying effects, foaming and solubilizing capacity, are studied. The influence of the degree of oligomerization and of the spacer group on the surfactant properties is discussed, in comparison with the analogous standard monomeric and dimeric ("gemini") surfactants. Typically, the evolution of the properties observed from standard to dimeric surfactants progresses with the trimers and tetramers, resulting for instance in extremely low critical micellization concentrations. PMID- 16042439 TI - Brush-type amphiphilic diblock copolymers from "living"/controlled radical polymerizations and their aggregation behavior. AB - Two brush-type amphiphilic diblock copolymers, poly(poly(ethylene glycol)methyl ether methacrylate-block-polystyrene) (P(PEGMA)-b-PS) and poly(glycidyl methacrylate)-block-poly(poly(ethylene glycol)methyl ether methacrylate) (P(GMA) b-P(PEGMA)) were synthesized, respectively, via consecutive atom-transfer radical polymerizations (ATRPs) and reversible addition-fragmentation chain-transfer (RAFT) polymerizations. The diblock copolymers were characterized by gel permeation chromatography (GPC), (1)H nuclear magnetic resonance (NMR) spectroscopy, and FT-IR spectroscopy. The aggregation behavior of the two amphiphilic diblock copolymers in water was also studied. Scanning electron and transmission electron microscopic images revealed that spherical micelles (40-80 nm in diameter) from self-assembly of the P(PEGMA)-b-PS copolymers and wormlike micelles (60-120 nm in length and 20-30 nm in diameter) from self-assembly of the P(GMA)-b-P(PEGMA) copolymers were prevalent. The spherical P(PEGMA)-b-PS micelles could self-assemble gradually into giant aggregates of several micrometers in diameter. PMID- 16042440 TI - Optical measurement of the deformation of giant lipid vesicles driven by a micropipet electrode. AB - We investigate the deformation of giant lipid vesicles driven by a micropipet electrode by use of differential confocal microscopy. This optical technique provides nanometer depth resolution without mechanical contact and hence prevents large tension or perforation of the soft membrane. For dipalmitoyl phosphatidylcholine (DPPC) membranes in the gel phase, we observed deformations of several hundreds of nanometers when the driving voltage was about 0.1 V. The voltage and frequency responses of the vesicle deformation can be explained by the balance between the electroosmotic force inside the micropipet and the membrane tension. We also used DPPC:cholesterol vesicles to check the validity of this model. In the fluid phase, however, the deformation is independent of the modulation signal because micrometer-scale thermal fluctuations dominate the membrane motion. PMID- 16042441 TI - Fibrous assemblies and water gelation in mixtures of lysine with sodium alkyl sulfates. AB - Thermoresponsive, pH-sensitive fibrous structures and gels are formed in aqueous mixtures of the amino acid lysine with oppositely charged sodium alkyl sulfate surfactants. The formation of these assemblies depends on the chain length of the surfactant, which is varied between 8 and 16, the chirality and degree of protonation of the amino acid, and the molar ratio of these species. Self assembly of the fibers occurs when specific lysine enantiomers are in solution and for pH conditions in which the majority of the amine groups are protonated (i.e., at near-equimolar amounts of HCl and lysine). Racemic mixtures of lysine do not form fibers with sodium dodecyl sulfate. Micelles are the fiber precursors, and the fibers, which can be hundreds of micrometers long, entangle to form gels. With increasing temperature, the gels melt, the fibers dissolve, and a single micellar phase forms. The micelles elongate with decreasing pH when the acid concentration is greater than equimolar with respect to lysine, and they shrink with increasing temperature. PMID- 16042442 TI - Assembly and mechanical properties of phosphorus dendrimer/polyelectrolyte multilayer microcapsules. AB - We report the preparation, characterization, and mechanical properties of polyelectrolyte/phosphorus dendrimer multilayer microcapsules. The shells of these microcapsules are composed either by alternating poly(styrenesulfonate) (PSS) and positively charged dendrimer G4(NH+Et2Cl-)96 or by alternating poly(allylamine hydrochloride) (PAH) and negatively charged dendrimer G4(CH-COO Na+)96. The same multilayers were constructed on planar support to examine their layer-by-layer growth and to measure the multilayer thickness. Surface plasmon resonance spectroscopy (SPR) showed regular linear growth of the assembly upon each bilayer deposited. We probe the mechanical properties of these polyelectrolyte/dendrimer microcapsules by measuring force-deformation curves with the atomic force microscope (AFM). The experiment suggests that they are much softer than PSS/PAH microcapsules studied before. This softening is attributed to an enhanced permeability of the polyelectrolyte/dendrimer multilayer shells as compared with multilayers formed by linear polyelectrolytes. In contrast, Young's modulus of both dendrimer-based multilayers was found to be on the same order as that of PSS/PAH multilayers. PMID- 16042443 TI - Formation of polysulfone colloids for adsorption of natural organic foulants. AB - An ever-present problem in the use of commercial membranes for treatment of drinking water is fouling of the membranes by natural organic matter (NOM). This work describes a new approach to elimination or minimization of membrane fouling by NOM. When a 2% solution of polysulfone in NMP and propionic acid is slowly injected into water, approximately 50 nm polysulfone particles are spontaneously formed, and these hydrophobic particles quickly coagulate into approximately 12 microm diameter aggregates; the formed material has a surface area of approximately 100 m(2)/g and an equivalent "pore" size of 25 nm. When 50 mg/L of the new material is equilibrated with a local drinking water supply, virtually all adsorptive fouling of a 20-kDa molecular weight cutoff ultrafiltration membrane is eliminated. Interestingly, although only a very small percentage of the NOM is removed by adsorption on the polysulfone aggregates, it appears that exactly this small NOM component is responsible for nearly all of the membrane fouling. This paper describes the fabrication and characterization of the new polysulfone adsorbent and offers an hypothesis for the formation of the product via spontaneous emulsification and spinodal decomposition. PMID- 16042444 TI - Cylindrical micelles of alpha-fluorocarbon-omega-hydrocarbon end-capped poly(N acylethylene imine)s. AB - Micelles of ABC block copolymers with varying degrees of polymerization of the B block (n) and constant lengths of the A and C blocks were investigated by small angle X-ray scattering (SAXS), analytical ultracentrifugation (AUC), surface tension measurements, and isothermal titration calorimetry. The copolymers consisting of hydrophilic poly(N-acylethylene imine)s, end-capped with a hydrophobic fluorocarbon and a hydrocarbon block, are polymeric surfactants (gamma = 35 mN/m). They form cylindrical micelles with radii of 3.0 nm (n = 35), 3.8 nm (n = 57), and 4.0 nm (n = 72). Their lengths are about 20 nm. The micelles can be doped with 1,4-diiodoperfluorobutane for the polymers with n = 57 and 72 but not for n = 35. We assume that the doped micelles form distinct fluorocarbon domains, which are able to incorporate selectively the fluorocarbon dopant. The work presented here is a contribution to the development of multicompartment micelles. PMID- 16042445 TI - Heavy-metal remediation by a fungus as a means of production of lead and cadmium carbonate crystals. AB - We show here that reaction of the fungus, Fusarium oxysporum, with the aqueous heavy-metal ions Pb2+ and Cd2+ results in the one-step formation of the corresponding metal carbonates. The metal carbonates are formed by reaction of the heavy-metal ions with CO2 produced by the fungus during metabolism and thus provide a completely biological method for production of crystals of metal carbonates. The PbCO3 and CdCO3 crystals thus produced have interesting morphologies that are shown to arise because of interaction of the growing crystals with specific proteins secreted by the fungus during reaction. An additional advantage of this approach is that the reaction leads to detoxification of the aqueous solution and could have immense potential for bioremediation of heavy metals. Under conditions of this study, the metal ions are not toxic to the fungus, which readily grows after exposure to the metal ions. PMID- 16042446 TI - STM study on quinacridone derivative assemblies: modulation of the two dimensional structure by coadsorption with dicarboxylic acids. AB - We describe the two-dimensional (2D) assemblies of N,N'-dialkyl-substituted quinacridone derivatives on highly orientated pyrolytic graphite observed by scanning tunneling microscopy, and focus our discussion on whether the supramolecular organization can be modulated by the coadsorption of dicarboxylic acids. Our experiments have demonstrated that the quinacridone derivatives can form different 2D nanostructures when coadsorbed with dicarboxylic acids of different length at the liquid/graphite interface. Interestingly, N,N' dihexadecyl-substituted quinacridone derivative alternately takes two different conformations in two columns for its coadsorption with pentadecanedioic acid and form a gridlike structure. It is shown that a cooperative effect of different interactions can be modulated by introducing guest molecule, leading to formation of different self-assembled nanostructures. PMID- 16042447 TI - Geometry-dependent stripe rearrangement processes induced by strain on preordered microwrinkle patterns. AB - A preordered microwrinkle pattern on a metal-capped surface of a soft elastomer is employed to elucidate the elementary buckling phenomenon during strain-induced stripe rearrangement processes. The preordered one-dimensional stripe tends to align perpendicular to the direction of strain reversibly when lateral compressive strain is applied on the substrate at some angle phi with respect to the stripe orientation. For any value of strain, the film surface can be decomposed in domains containing stripes with two different orientations, namely the original and applied strain orientations. As strain is increased, the domains of the second type of stripes progressively grow and invade the whole surface. Interestingly, the domain shapes during growth are composed of parallelogram units that simply depend on phi and stripe wavelength. Moreover, domain growth proceeds in characteristic directions depending on the shape of the domain unit. PMID- 16042448 TI - Intercalation of gaseous thiols and sulfides into Ag+ ion-exchanged aluminum dihydrogen triphosphate. AB - Ag(+) ion-exchanged layered aluminum dihydrogen triphosphate (AlP) with the interlayer distance of 0.85 nm was synthesized by the ion-exchange of proton in triphosphate with Ag(+) ion. The amount of exchanged Ag(+) ion depended on the concentration of AgNO(3) aqueous solution. Ag(+) ion-exchanged AlP adsorbed gaseous thiols and sulfides into the interlayer region. The adsorption amounts of thiols were more than those of sulfides, thiols with one mercapto group > thiol with two mercapto groups > sulfides, and depended on the amount of exchanged Ag(+) ion in the interlayer region. The thiols with one mercapto group were intercalated to expand the interlayer distance of Ag(+) ion-exchanged AlP, whereas there was no expansion in the adsorption of sulfide. In the case of thiol with two mercapto groups, there was observed contraction of the interlayer distance through the bridging with Ag(+) ions of the upper and lower sides of the interlayer region. PMID- 16042449 TI - Modeling of surface heterogeneity of microporous adsorbents with LBET approach. AB - A package of new isotherm equations aimed at examination of pore structure of sub and microporous materials with respect to a surface heterogeneity is proposed. One considers adsorption of small nearly spherical molecules in irregular pores of molecular size. A generalized BET theory is exploited together with a thermodynamic description of the process and handling restrictions for multilayer adsorption (LBET approach). Realistic multivariant relationships linking a surface energy distribution with the pore size are proposed, and exponential distribution of the pore size is accepted. As a result, the heterogeneous adsorption equations (uLBET formulas) are derived, dedicated for irregular microporous systems. It was shown that they may be well-approximated with analytical BET-like formulas (the models of the LBET-type), enabling fast multivariant examination of real microporous materials. Such an examination provides complementary information on adsorption mechanisms, thus, allowing more reliable evaluation of the system parameters. Results of the new model's application to an empirical system (nitrogen adsorption on an activated carbon) are shown. PMID- 16042450 TI - Mixtures of sodium dodecyl sulfate/dodecanol at the air/water interface by computer simulations. AB - Molecular dynamics simulations of monolayers of surfactant mixtures at the air/water interface were performed where the binary mixture was composed of sodium dodecyl sulfate (SDS) and dodecanol molecules. At the same ratio of SDS and dodecanol molecules, two monolayer mixtures were prepared. In the first monolayer, all the dodecanol molecules were placed together in the center of the simulation box, whereas in the second monolayer, those molecules were uniformly distributed in the surface area in such a way that they were far from each other. Simulations of both systems indicate that the dodecanol tails in the first monolayer are straighter and more ordered than those in the second monolayer. From the present results, we observed new insights of how the different molecules should array or distribute at the interface in real systems. Finally, studies of the interfacial water around the different surfactants were also analyzed, showing that they are closer to the polar headgroups of dodecanol than to the SDS headgroups. PMID- 16042451 TI - Nanoscale experimental investigation of particle interactions at the origin of the cohesion of cement. AB - Atomic force microscopy has been used to investigate the force at the origin of the cohesion of cement. The cohesion of cement grains is caused by surface forces acting between calcium silicate hydrate nanoparticles in interstitial electrolytic solution. Direct measurement of the interaction between two calcium silicate hydrate surfaces is performed in air and different aqueous solutions. In dry air, starting with the van der Waals forces, the interaction area between calcium silicate hydrate nanoparticles can be estimated. In electrolytic solution, the evolution of these forces is extensively dependent on both surface and solution chemistry. The roles of the calcium hydroxide concentration, pH, and ionic strength are investigated. The force measurements allow us to confirm the pre-eminence of ionic correlation forces in the cohesion of cement. PMID- 16042452 TI - Colloidal rings in a liquid mixture. AB - We investigate the self-assembly of colloidal particles on microscopic decane droplets in water and show that, by use of paramagnetic colloids, it is possible to assemble ringlike structures that can be controlled with a magnetic field. Moreover, the use of paramagnetic colloids allows us to determine the attractive forces between the colloids located at the three-phase contact line between decane, water, and air. The attractive force is in the femtonewton range and is attributed to capillary interactions due to interface deformations. When the liquid emulsion dries on a glass slide, we observe solid deposits in the form of microscopic rings of varying diameters. PMID- 16042453 TI - Self-assembly of colloidal pyramids in magnetic fields. AB - We study routes toward the construction of 2D colloidal pyramids. We find that magnetic beads may self-assemble into pyramids near a nonmagnetic 1D boundary as long as the number of beads in the pyramid does not exceed 10. We have also found that a strong magnetic field gradient could act as a boundary, thus assisting the self-assembly of magnetic colloids in water, and have observed the formation of stable microscopic pyramids within a certain magnetic field range. Our results indicate that colloidal pyramids can be formed in a number of ways by utilizing external fields. PMID- 16042454 TI - Selective extraction and recovery of cytochrome c by liquid-liquid extraction using a calix[6]arene carboxylic acid derivative. AB - Recently, we reported that a calix[6]arene carboxylic acid derivative can selectively extract the lysine-rich protein cytochrome c by interacting with amino groups on the protein surface. In the present article, quantitative extraction and recovery of cytochrome c using this calix[6]arene carboxylic acid derivative are described. Both adjustment of the pH under acidic conditions and addition of an alcohol are necessary to strip the extracted protein from an organic solution to an aqueous solution. Separation of cytochrome c and lysozyme using the calix[6]arene was achieved under the optimal conditions. In the forward extraction stage, 93% of the cytochrome c was extracted, while lysozyme remained in the solution. In the subsequent stripping stage, the extracted cytochrome c was quantitatively recovered in an aqueous solution. Finally, separation of these proteins, which have similar molecular weights and isoelectric points, was accomplished. PMID- 16042455 TI - Scanning tunneling microscopy investigation of the structure of methanethiolate on Ag(111). AB - Scanning tunneling microscopy (STM) has been used to investigate the structure of the ordered methanethiolate overlayer formed on Ag(111) by reaction at room temperature with dimethyl disulfide. High-resolution images show an ordered structure with three inequivalent atomic-scale protrusions within each ( radical7 x radical7)R19 degrees surface unit mesh which can be reconciled with methanethiolate species on a regular lateral submesh, similar to that proposed in the multilayer ( radical7 x radical7)R19 degrees -S sulfide phase previously reported. STM imaging during dynamic dosing also provides evidence for a significant change in the outermost layer Ag atom density, consistent with a reconstructed surface model. Possible models for this reconstruction are presented and discussed in the light of available information. PMID- 16042456 TI - Hydrogels from a water-soluble zwitterionic polythiophene: dynamics under pH change and biomolecular interactions observed using quartz crystal microbalance with dissipation monitoring. AB - The water-soluble zwitterionic polythiophene, poly(3-((S)-5-amino-5-carboxyl-3 oxapentyl)-2,5-thiophene) hydrochloride (POWT), is a conjugated polyelectrolyte (CPE) with properties well suited for biochip applications. CPEs readily form hydrogels when exposed to water-based buffer solutions or biomolecule solutions. In this work, we used in situ quartz crystal microbalance with dissipation (QCM D) monitoring to collect information on the interaction between POWT films exposed to buffers with different pH and POWT/DNA chains. Our data show that POWT swells significantly when exposed to low-pH buffers, such as pH 4 acetate, this is seen as an increase in thickness and decrease in viscosity obtained via a Voight-based modeling of combined f and D QCM-D measurements. The magnitude of thickness and viscosity change upon changing from a pH 10 carbonate buffer to pH 4 acetate is 100% increase in thickness and 50% decrease in viscosity. The response of the hydrogel under pH change is well correlated with fluorescence data from POWT films on glass. The state of the hydrogel is important during interaction with biomolecules; illustrated by the observation that a swollen CPE hydrogel adsorbs a higher amount of DNA than a compacted one. In agreement with previous results, the QCM-D data confirmed that the POWT/DNA hydrogel sense complementary DNA specifically and with negligible binding of noncomplementary DNA. These results are important for efficient constructions of biochips in water environments using this class of materials. PMID- 16042457 TI - Super-liquid-repellent surfaces prepared by colloidal silica nanoparticles covered with fluoroalkyl groups. AB - A simple and easy method to prepare super-liquid-repellent surfaces is proposed. Sol-gel films were prepared by hydrolysis and condensation of alkoxysilane compounds. Both surface energy and roughness were controlled using colloidal silica particles and fluoroalkylsilane. When the fractional amounts of both colloidal silica and fluoroalkylsilane were optimized in the films, the film surface exhibited repellency to both water and oil. Finally, it was shown that the method proposed here would be applied to a simple one-pot coating for a uniform large area, and be useful for practical use. PMID- 16042458 TI - Kinetics for the collapse of trilayer liquid-crystalline disks from a monolayer at an air-water interface. AB - Unlike surfactants considered in previous studies, when phosphatidylcholine (PC) monolayers collapse at constant surface tension to form a 3D bulk phase, surface area decreases at rates that slow. The different kinetics could result from collapse by a distinct mechanism. Rather than the transfer of molecules all along the interface between the monolayer and bulk phase, PC films can collapse by the folding and subsequent sliding of a bilayer over the monolayer. By this mechanism, molecules can transfer to collapsed trilayers through a locus of constant size. In this article, we use the theory of nucleation and growth to show analytically that during collapse, the area can decrease at rates that decelerate when each individual structure grows through a region of fixed dimensions. We also show that binary films of 30% dihydrocholesterol (dchol) and dipalmitoyl phosphatidylcholine (DPPC), which have previously been shown to form a homogeneous monolayer from which trilayer disks grow through a point, collapse with rates of area decay that slow, in agreement with our analytical expressions. PMID- 16042459 TI - Evaporation from water-ethylene glycol liquid mixture. AB - Evaporation rates were determined for water-ethylene glycol liquid mixtures with different mole fractions, where the evaporation rate expressed as mg min(-1)/area was used because of the presence of two kinds of molecular species. The rate increased with increasing temperature and decreased with increasing mole fraction of ethylene glycol, almost obeying ideal mixing of the two components, although a small positive deviation was observed over the mole fraction from 0 to 0.5 of ethylene glycol at higher temperatures. The activation energy of evaporation was determined from the temperature dependence of the evaporation rate, where the energy was an apparent one because the composition of evaporated species was not determined. The activation energy increased with decreasing temperature and with increasing mole fraction of ethylene glycol, where the energy obeyed the ideal mixing at lower temperatures while it positively deviated at higher temperatures. The evaporation rates were examined by surface tension of the liquid mixture, but any definite relation between them was not found. Both the evaporation rate and the activation energy were found to be determined mainly by the mole fraction in the surface layer from which the evaporation takes place. Finally, the new concept of surface excess was presented, where the surfactant molecules were concentrated and formed a bimolecular layer at a certain distance beneath the air/solution interface. PMID- 16042460 TI - Thermodesorption studies of energetic properties of Ni/MgO-Al2O3 catalysts. Determination of adsorption energy distribution functions. AB - The thermodesorption spectra of hydrogen from coprecipitated catalysts (70-x)NiO xMgO-30Al(2)O(3) (x = 0-50%(wt)) are reported. The catalysts were calcined at 400 degrees C and reduced with H(2) at 20-800 degrees C and for 3 h at 800 degrees C. NiO reduction degree was between 49.3 and 92.1%. The active surface areas changed from 8.4 to 32.4 m(2)/g whereas mean size of nickel crystallites was between 3.7 and 9.7 nm. The TPD spectra were next analyzed in order to determine the adsorption energy distributions functions. To obtain these functions a theoretical model of adsorption/desorption kinetics based on the statistical rate theory (SRT) was applied. This approach allows for determination of the adsorption energy at nonequilibrium conditions as well as at quasiequilibrium conditions. The resulting distribution functions reveal the presence of two main bands of adsorption energy. Some correlation is found between the determined distributions of adsorption energy and the size of nickel crystallites determined using the XRD method. The presence of MgO favors creation of high energy adsorption sites on Ni crystallites. PMID- 16042461 TI - In situ quadrupole mass spectrometry study of atomic-layer deposition of ZrO2 using Cp2Zr(CH3)2 and water. AB - Reactions during the atomic layer deposition (ALD) process of ZrO(2) from Cp(2)Zr(CH(3))(2) and deuterated water as precursors were studied with a quadrupole mass spectrometer (QMS) at 210-440 degrees C. The detected reaction byproducts were CpD (m/z = 67) and CH(3)D (m/z = 17). Almost all (90%) of the CH(3) ligands were released during the Cp(2)Zr(CH(3))(2) precursor pulse because of exchange reactions with the OD-terminated surface, and the rest, during the D(2)O pulse. About 40% of the CpD was released during the metal precursor pulse, and 60%, during the D(2)O pulse. ALD-type self-limiting growth was confirmed from 210 to 400 degrees C. However, below 300 degrees C the growth rate was low. Precursor decomposition affected the film growth mechanism at temperatures exceeding 400 degrees C. PMID- 16042462 TI - Study of pH-sensitive copolymer/phospholipid complexes using the langmuir balance technique: effect of anchoring sequence and copolymer molecular weight. AB - The behavior of three copolymers of N-isopropylacrylamide (NIPAM), methacrylic acid (MAA), and hydrophobic moiety was studied at phospholipid monolayer/subphase interfaces. The hydrophobic moieties, N-terminal dioctadecylamine (DODA) and random octadecylacrylate (ODA), were used as anchoring groups. The interactions between a 1,2-distearoyl-sn-glycero-3-phosphatidylcholine (DSPC) monolayer and the copolymers were studied using the Langmuir balance technique. The effect of subphase pH, distribution of anchors along the copolymer chain, and copolymer molecular weight on the nature of the interactions between the copolymer chains and the DSPC monolayer were investigated. A first-order kinetics model was used to analyze the copolymers adsorption at the DSPC monolayer/subphase interface and allowed the interaction area between the copolymer chains and the DSPC monolayer, A(x), to be determined. The interaction area appears to depend on the subphase pH and the copolymer molecular weight. On decreasing pH, the interaction area of high molecular weight copolymers increases significantly; this is consistent with the copolymer chain phase transition from an extended coil to a collapsed globule while pH is lowered. In the latter conformation, strong hydrophobic attractive interactions between the copolymer chains and the hydrophobic part of the DSPC monolayer favor the copolymer intercalation, which could eventually provoke the phospholipidic layer destabilization or rupture. PMID- 16042463 TI - Modeling of binary adsorption on heterogeneous surfaces characterized by a quasi gaussian adsorption energy distribution. AB - The integral equation (IE) approach coupled with a quasi-Gaussian adsorption energy distribution is used to model the adsorption of single gases and their binary mixture on a heterogeneous solid surface. The adsorbing surface is assumed to be characterized by two, generally different in width, quasi-Gaussian distribution functions, each of them related to a single component of the mixture. The influence of correlations between the distribution functions associated with different components on the corresponding adsorption isotherms and phase diagrams is discussed. In particular, it is demonstrated that a lack of microscopic correlations between the adsorption energies of the components may lead to the formation of an azeotropic mixture. The predictions of the theory are also compared with the results of the grand canonical Monte Carlo (GCMC) simulations carried out for the system studied. PMID- 16042464 TI - Combined surface pressure-interfacial shear rheology study of the effect of pH on the adsorption of proteins at the air-water interface. AB - The effect of pH on the adsorption of catalase and lysozyme at the air-water interface has been studied using a combined surface pressure-interfacial shear rheology technique. The results presented show that the rate of development of interfacial phenomena increases as the pH of the subphase approaches the isoelectric point of the protein under investigation. The development of the measured interfacial rheological parameters is due to an increased rate of cross link formation within the resultant interfacial gel. The formation of the interfacial gels has been modeled using a combination of the Smoluchowski theory for the coagulation of an aerosol or fog and classic rubber elasticity theory. The enhanced rate of cross-link formation at the isoelectric point is a result of an in-surface phase separation brought about by cooperative deionization of the protein molecules near their isoelectric point. Simultaneous measurements of surface pressure and interfacial rheology have enabled us to show that the development of a gel-like interfacial network coincides with observed increases in surface pressure. PMID- 16042465 TI - Thermodynamic and structural properties of phospholipid langmuir monolayers on hydrosol surfaces. AB - Measurements of Langmuir pressure/area isotherms, rheology, grazing incidence X ray diffraction (GIXD), and grazing incidence diffuse X-ray scattering out of the specular plane (GIXOS) have been used to investigate the influence of a hydrosol containing charged mineral nanoparticles on the thermodynamic and structural properties of a DPPC monolayer. The mineral adsorption layer that is formed via electrostatic interaction underneath the lipid layer alters the thermodynamic properties of the phospholipid monolayer in terms of maximal achievable compression, compressibility, and phase behavior. Modifications appear in the latter case as a coolinglike effect. Rheology measurements of the bulk viscoelastic properties revealed a stabilizing effect of the transient bulk network on the surfactant layer. The lipid chain lattice is found to be reorganized and adapted to the internal atomic structure of the mineral particles. A model for the superposition of Bragg rods from the lipid chains and the minerals is applied to separate these scattering contributions. In the vicinity of the mineral particles, the (2) reflection for DPPC on a liquid substrate was found, indicating strongly suppressed fluctuations at the surface. An estimation of the Debye-Waller factor associated with the lipid layer organization is used to quantify the damping of fluctuations within the lipid matrix due to the rigidifying and stabilizing effect of the mineral particles. PMID- 16042466 TI - Nanometer-thick surficial films in oxides as a case of prewetting. AB - Stable, nanometer-thick films are observed to form at the {1120} facets of Bi(2)O(3)-doped ZnO in several bulk-phase stability fields. Electron microscopy shows these surficial films to exhibit some degree of partial order in quenched samples. The equilibrium film thickness, corresponding to the Gibbs excess solute, decreases monotonically with decreasing temperature until vanishing at a dewetting temperature, well below the eutectic. Assuming that perfect wetting occurs at some higher temperature above the eutectic, as is observed on polycrystal surfaces and at grain boundaries in the same system, the adsorption and wetting events in this system illustrate temperature- and composition dependent prewetting. The observation of a second class of thicker films coexisting with nanodroplets and a numerical evaluation of thickness versus temperature elucidate the critical role of volumetric thermodynamic terms in determining film stability and thickness. Analogous temperature-dependent surface films involving adsorbed MoO(3) on Al(2)O(3) were also observed. PMID- 16042467 TI - Application of percolation theory to the drainage of liquid nitrogen from mesoporous silica xerogel Gelsil 50. AB - Previous in situ small-angle neutron scattering studies of nitrogen adsorption and desorption at 78 K on the mesoporous silica xerogels Gelsil 50 and Gelsil 75 revealed the formation of ramified clusters of vapor-filled pores on desorption, which is characteristic for a percolation process. In the present work, we check whether the adsorption/desorption isotherm data for a monolithic sample of Gelsil 50 can be analyzed in terms of a bond-percolation model. Three powder samples were studied too. Percolation probability data are presented and the effects of heterogeneous nucleation, finite size, and surface clusters on drainage from Gelsil 50 are addressed. The mean coordination number was derived. The results of the analysis are discussed with respect to recent theoretical work for interactions of fluids with complex pore systems. The monolithic sample and a powder sample were characterized by small-angle neutron scattering data. PMID- 16042468 TI - Study of frictional properties of a phospholipid bilayer in a liquid environment with lateral force microscopy as a function of NaCl concentration. AB - Friction properties of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) supported planar bilayers deposited on mica were tested in a liquid environment by lateral force microscopy. The presence of these bilayers was detected by imaging and force measurements with atomic force microscopy. To test how the presence of NaCl affects the frictional properties of the phospholipid bilayers, four DMPC bilayers were prepared on mica in saline media ranging from 0 to 0.1 M NaCl. Changes in the lateral vs vertical force curves were recorded as a function of NaCl concentration and related to structural changes induced in the DMPC bilayer by electrolyte ions. Three friction regimes were observed as the vertical force exerted by the tip on the bilayer increased. To relate the friction response to the structure of the DMPC bilayer, topographic images were recorded at the same time as friction data. Ions in solution screened charges present in DMPC polar heads, leading to more compact bilayers. As a consequence, the vertical force at which the bilayer broke during friction experiments increased with NaCl concentration. In addition, the topographic images showed that low-NaCl concentration bilayers recover more easily due to the low cohesion between phospholipid molecules. PMID- 16042469 TI - Polystyrene-block-poly(ethylene oxide) stars as surface films at the air/water interface. AB - Star diblock copolymers containing polystyrene (PS) and poly(ethylene oxide) (PEO) were investigated as surface films at the air/water interface. Both classic and dendritic-like stars were prepared containing either a PS core and PEO corona or the reverse. The investigated polymers, consisting of systematic variations in architectures and compositions, were spread at the air/water interface, generating reproducible surface pressure-area isotherms. All of the films could be compressed to higher pressures than would be possible for pure PEO. For stars containing 20% or more PEO, three distinct regions appeared. At higher areas, the PEO absorbs in pancakelike structures at the interface with PS globules sitting atop. Upon compression, a pseudoplateau transition region appeared. Both regions strongly depended on PEO composition. The pancake area and the pseudoplateau width and pressure increased in a linear fashion with an increasing amount of PEO. In addition, minimum limits of PEO chain length and mass percentage were determined for observing a pseudoplateau. At small areas, the film proved less compressible, producing a rigid film in which PS dominated. Here, the film area increased with both molecular weight and the amount of PS. Comparison with pure linear PS showed the stars spread more, occupying greater areas. Among the stars, the PEO-core stars were more compact while the PS-core stars spread more. The influence of architecture in terms of the core/corona polymers and branching were also examined. The effects of architecture were subtle, proving less important than PEO chain length or mass percentage. PMID- 16042470 TI - Adsorption of liquid mixtures on silicalite-1 zeolite: a density-bottle method. AB - A technique that measures the effective density of a zeolite after adsorption from the liquid phase was developed to measure the absolute amounts of liquid mixtures adsorbed on zeolites without using a nonadsorbing solvent. Since the fugacities of the adsorbing components in solution can be dramatically different with or without the addition of a nonadsorbing solvent, this technique measures mixture isotherms that can be used for analyzing pervaporation through zeolite membranes. A nonideal solution, methanol/acetone, was used as an example to show that its adsorption isotherms on silicalite-1 zeolite at 294 K differ dramatically from those measured with the nonadsorbing solvent method. The methanol/acetone fugacity ratio is different for the two methods because of different concentrations in the liquid phase. Methanol preferentially adsorbs on silicalite-1 at low methanol concentrations and acetone preferentially adsorbs at high methanol concentrations. The density bottle method was used to show that n hexane preferentially adsorbs from n-hexane/3-methylpentane liquid mixtures, and at high n-hexane concentrations, essentially no 3-methylpentane adsorbs, as has been predicted previously by simulations. A larger molecule, 2,2-dimethylbutane, adsorbed so slowly at 294 K that silicalite had only 16% of saturation coverage after 370 h, but it was saturated after 1650 h; at 423 K, saturation was obtained in less than 24 h. PMID- 16042471 TI - Selective determination of surface density of bromide ion through XAFS and its application to verification of a criterion of an ideal mixing of surfactant mixture. AB - The mole fraction of chloride ion of dodecyltrimethylammonium bromide (DTAB) and dodecyltrimethylammonium chloride (DTAC) mixture in the adsorbed film XHC was estimated not only by the thermodynamic analysis of the surface tension data but also by analyzing the Br K-edge jump of the XAFS spectrum under the total reflection condition (TRXAFS method). The phase diagrams of adsorption (PDA) at several surface tensions from the two methods were in good agreement. On the basis of the PDA obtained, it was clearly shown that the criterion of an ideal mixing for the DTAB-DTAC system is not given by the linear relation between the total molality of surfactant mixture m and XHC, m = m0B + (m0C - m0B)XHC, but by the one between m2 and XHC, m2 = (m0B)2 + [(m0C)2 - (m0B)2]XHC. Furthermore, it was demonstrated that the theoretical approach that provides the latter relation draws a distinction between the criteria for an ionic surfactant mixture without a common ion and that for an ionic surfactant mixture with a common ion. PMID- 16042472 TI - Structure and stability of silica particle monolayers at horizontal and vertical octane-water interfaces. AB - Monolayers of silica particles at horizontal and vertical octane-water interfaces have been studied by microscopy. It is found that their structure and stability depend strongly on the particle hydrophobicity. Very hydrophobic silica particles, with a contact angle of 152 degrees measured through the water, give well-ordered monolayers at interparticle distances larger than 5 particle diameters which are stable toward aggregation and sedimentation. In contrast, monolayers of less-hydrophobic particles are disordered and unstable. Two dimensional particle sedimentation has been observed in the case of vertical monolayers. The results have been analyzed with a simple two-particle model considering the sedimentation equilibrium as a balance between the long-range electrostatic repulsion through the oil, the gravity force, and the capillary attraction due to deformation of the fluid interface around particles. The value of the charge density at the particle-octane interface, 14.1 muC/m(2), found for the most hydrophobic particles is reasonable. It drastically decreases for particles with lower hydrophobicity, which is consistent with the order-disorder transition in monolayer structure reported by us before. The pair interactions between particles at a horizontal octane-water interface have been analyzed including the capillary attraction due to undulated three-phase contact line caused by nonuniform wetting (the contact angle hysteresis). The results are in agreement with the great stability of very hydrophobic silica particle monolayers detected experimentally, even at low pH at the point of zero charge of the particle-water interface, and with the aggregated structure of hydrophilic particle monolayers. PMID- 16042473 TI - Nondestructive monitoring of the photochromic state of dithienylethene monolayers by surface plasmon resonance. AB - The use of surface plasmon resonance (SPR) as a nondestructive, nonerasing readout of the isomerization state of a photochromic dithienylethene covalently linked to a chemically modified gold surface was investigated. Four different binding layers were examined: 11-mercaptoundecanol (MUO), an amine-modified 11 mercaptoundecanol (MUO-NH2), dextran, and an amine-modified dextran. The binding of dithienylethene to the modified gold surface and photoisomerization of the photochrome in the bound state were established by FTIR. Solvent effects were measured for every layer tested using ethanol and hexanes. In general, large, easily measurable SPR signal changes could be detected under conditions where photoisomerization of the dithienylethene photochrome was not quenched by the gold plasmon, establishing SPR as a viable form of readout for potential dithienylethene-based optical data storage or processing devices. Dextran-bound photochrome in ethanol exhibited the largest SPR response upon photoisomerization, but is more prone to time-dependent fluctuations resulting from swelling of the dextran layer (caused by slow diffusion of the solvent) than the other layers. Large responses are also provided by MUO-NH2 and MUO, and the signal is much more stable than that for dextran. PMID- 16042474 TI - The surface charging at low density of protonatable surface sites. AB - The point of zero charge (PZC) of a sparingly soluble metal oxide depends on the density of protonatable surface oxygen atoms. The shift in the PZC is due to protonation/deprotonation of water in the regions free of protonatable surface oxygen atoms originating from the solid. The PZC of alumina increases when the density of protonatable surface oxygen atoms increases. In contrast, the PZC of titania is rather insensitive to the density of protonatable surface oxygen atoms. In surfaces of many materials the regions free of protonatable surface oxygen atoms dominate. These materials have a PZC at pH about 4. PMID- 16042475 TI - Partial dewetting of polyethylene thin films on rough silicon dioxide surfaces. AB - The effect of roughness on the dewetting behavior of polyethylene thin films on silicon dioxide substrates is presented. Smooth and rough silicon dioxide substrates of 0.3 and 3.2-3.9 nm root-mean-square roughness were prepared by thermal oxidation of silicon wafers and plasma-enhanced chemical vapor deposition on silicon wafers, respectively. Polymer thin films of approximately 80 nm thickness were deposited by spin-coating on these substrates. Subsequent dewetting and crystallization of the polyethylene were observed by hot-stage optical microscopy in reflection mode. During heating, the polymer films melt and dewet on both substrates. Further observations after cooling indicate that, whereas complete dewetting occurs on the smooth substrate surface, partial dewetting occurs for the polymer film on the rough surface. The average thickness of the residual film on the rough surface was determined by ellipsometry to be a few nanometers, and the spatial distribution of the polymer in the cavities of the rough surface could be obtained by X-ray reflectometry. The residual film originates from the impregnation of the porous surface by the polymer fluid, leading to the observed partial dewetting behavior. This new type of partial dewetting should have important practical consequences, as most real surfaces exhibit significant roughness. PMID- 16042476 TI - Measurement of the streaming potential and streaming current near a rotating disk to determine its zeta potential. AB - Methodology for determining the zeta potential of a disk-shaped sample by both streaming potential and streaming current measurements is presented. Integration of Laplace's equation within one radius of the disk surface revealed that the streaming potential decreased strongly in the surface normal direction. With this solution, the zeta potential can be calculated from measurements of the streaming potential near the surface of the disk provided the position of the working electrode near the disk surface is known. Determining the zeta potential of a disk-shaped sample by means of streaming current measurements required determination of a current collection efficiency because not all the streaming current from a disk flows through the auxiliary electronic current path. While the working electrode near the disk should be pointlike, several possible variants on counter electrode shape and size were explored. Although the current collection efficiency was only a few percent in each case, the measured current was of 10 nA order. The current collection efficiency depended only on system geometry and was independent of a disk's zeta potential and solution concentration. Streaming current measurements of zeta potential on silicon wafers in potassium chloride solutions up to 10 mM agreed well with published values. PMID- 16042477 TI - Preparation and electrochemical behavior of ordered rh adlayers on Pt(100) electrodes. AB - Rhodium adlayers on Pt(100) substrates have been prepared by electrodeposition from dilute Rh(III) acidic solutions. The initially disordered layer is electrochemically annealed by applying a polarization program consisting of high sweep-rate multicycle sequences between 0.05 and 0.78 V(RHE) in 0.1 M H(2)SO(4). In this way, a pseudomorphic Rh monolayer can be prepared on Pt(100) substrates. The degree of order of the electrochemically annealed layer has been evidenced not only through voltammetric experiments but also by means of scanning tunneling microscopy with atomic resolution for iodine-protected adlayers, which show a c(2 x 2) structure. The electrochemically induced ordering of the Rh adlayer appears to be a consequence of the repeated cycles of adsorption/desorption of H and, especially, oxygenated species. Voltammetry in sulfuric acid solutions permits examination of the energetics of H/anions and OH/O adsorption as a function of the Rh coverage. The first monolayer adsorbs both hydrogen and oxygenated species more strongly than the second one. This can be explained through an electronic effect caused by the underlying Pt(100) substrate. PMID- 16042478 TI - Electrochemical and surfaced-enhanced Raman spectroscopic investigation of CO and SCN- adsorbed on Au(core)-Pt(shell) nanoparticles supported on GC electrodes. AB - Core-shell Au-Pt nanoparticles were synthesized by using a seed growth method and characterized by transmission electron microscopy, X-ray diffraction, and UV-vis spectroscopy. Au(core)-Pt(shell)/GC electrodes were prepared by drop-coating the nanoparticles on clean glassy carbon (GC) surfaces, and their electrochemical behavior in 0.5 M H2SO4 revealed that coating of the Au core by the Pt shell is complete. The electrooxidation of carbon monoxide and methanol on the Au(core) Pt(shell)/GC was also examined, and the results are similar to those obtained on a bulk Pt electrode. High quality surface-enhanced Raman scattering (SERS) spectra of both adsorbed CO and thiocyanate were observed on the Au(core) Pt(shell)/GC electrodes. The potential-dependent SERS features resemble those obtained on electrochemically roughened bulk Pt or Pt thin films deposited on roughened Au electrodes. For thiocyanate, the C-N stretching frequency increases with the applied potential, yielding two distinctly different dnu(CN)/dE. From 0.8 to -0.2 V, the dnu(CN)/dE is ca. 50 cm(-1)/V, whereas it is 90 cm(-1)/V above 0 V. The bandwidth along with the band intensity increases sharply above 0 V. At the low-frequency region, Pt-NCS stretching mode at 350 cm(-1) was observed at the potentials from -0.8 to 0 V, whereas the Pt-SCN mode at 280 cm(-1) was largely absent until around 0 V and became dominant at more positive potentials. These potential-dependent spectral transitions were attributed to the adsorption orientation switch from N-bound dominant at the negative potential region to S bound at more positive potentials. The origin of the SERS activity of the particles is briefly discussed. The study demonstrates a new method of obtaining high quality SERS on Pt-group transition metals, with the possibility of tuning SERS activity by varying the core size and the shell thickness. PMID- 16042479 TI - Direct electrochemistry and Raman spectroscopy of sol-gel-encapsulated myoglobin. AB - The direct electrochemistry of myoglobin (Mb) has been observed at a glassy carbon (GC) electrode coated with silica sol-gel-encapsulated Mb film. A well behaved cyclic voltammogram is observed with a midpoint potential (E(1/2)) of 0.25 V vs Ag/AgCl in a pH 7.0 phosphate buffer. This potential, which is pH dependent, is 70-90 mV more negative than the formal potential values obtained by using the spectroeletrochemical titration method at the same pH. Square wave voltametry (SWV) also shows a peak potential of -0.25 V for the reduction of Mb under the same experimental conditions. Both cathodic and anodic peak currents have a linear relationship with the scan rate. The midpoint potential decreases with pH, having a slope of -30 mV/pH. UV-vis and resonance Raman spectroscopic studies reveal that the sol-gel provides a bio-compatible environment where Mb retains a structure similar to its solution form, a 6-coordinated aquomet myoglobin. These results suggest that the silica sol-gel is a useful matrix for studying direct electrochemistry of other heme proteins. PMID- 16042480 TI - Electrochemical impedance spectroscopy of polyelectrolyte multilayer modified gold electrodes: influence of supporting electrolyte and temperature. AB - Electrochemical impedance spectroscopy and cyclic voltammetry are employed to characterize poly(styrenesulfonate)/poly(allylamine hydrochloride) multilayers assembled onto cysteamine-modified gold surfaces. The influence of the supporting electrolyte and temperature on the impedance response is studied because of both its practical interest and the need to test further the capillary membrane model recently developed by Barreira et al. [J. Phys. Chem. B 2004, 108, 17973]. The results obtained are interpreted quite satisfactorily in terms of this model, thus providing additional support to its usefulness for the description of ionic transport through polyelectrolyte multilayers. It is observed that the nature of the supporting electrolyte affects the film resistance and the electrode coverage. The temperature dependence of the diffusion coefficient is shown to follow the Arrhenius law, and the activation energy is estimated as 61 kJ/mol. Experiments with a large number of layers are also included to show that the impedance response of the multilayer then resembles that of a homogeneous membrane. PMID- 16042481 TI - Patterned biomimetic membranes: effect of concentration and pH. AB - Planar-supported lipid bilayers have attracted enormous attention because of their properties as model cell membranes, which can be employed in a variety of fundamental biological studies and medical devices. Furthermore, the development of patterned biological interfaces is of great practical and scientific interest because of their potential applications in the field of biosensors, drug screening, tissue engineering, and medical implants. In this study, mica supported membranes were constructed from biomimetic peptide-amphiphiles and their mixtures with lipidated poly(ethylene glycol) (PEG120) molecules or 1,2 dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) phospholipids using the Langmuir Blodgett technique. The two peptide-amphiphiles used in this study were a fibronectin-mimetic with the PHSRN(SG)(3)SGRGDSP headgroup (referred to as PHSRN GRGDSP) that contains both the primary (GRGDSP) and the synergy (PHSRN) recognition sites for alpha(5)beta(1) integrins and a peptide-amphiphile that mimics a fragment of the N-terminus of the fractalkine receptor (referred to as NTFR). Compression isotherms of the peptide-amphiphiles and their mixtures with PEG120 at the air/water interface were recorded and analyzed to evaluate the extent of miscibility in the two-component LB films. Domain formation in mica supported bilayers constructed from mixtures of peptide-amphiphiles and lipidated PEG120 or DPPC was observed using atomic force microscopy. In PHSRN-GRGDSP/PEG120 mixtures deposited from an aqueous subphase at pH 7, concentration-dependent phase separation was observed on the AFM images. The NTFR/PEG120 and NTFR/DPPC mixtures deposited at pH 10 exhibited extensive lateral phase separation at all mixture compositions, whereas at deposition pH 7 the concentrations of NTFR/DPPC examined here were well mixed. PMID- 16042482 TI - Fluid and air-stable lipopolymer membranes for biosensor applications. AB - The behavior of poly(ethylene glycol) (PEG) conjugated lipids was investigated in planar supported egg phosphatidylcholine bilayers as a function of lipopolymer density, chain length of the PEG moiety, and type of alkyl chains on the PEG lipid. Fluorescence recovery after photobleaching measurements verified that dye labeled lipids in the membrane as well as the lipopolymer itself maintained a substantial degree of fluidity under most conditions that were investigated. PEG densities exceeding the onset of the mushroom-to-brush phase transition were found to confer air stability to the supported membrane. On the other hand, substantial damage or complete delamination of the lipid bilayer was observed at lower polymer densities. The presence of PEG in the membrane did not substantially hinder the binding of streptavidin to biotinylated lipids present in the bilayer. Furthermore, above the onset of the transition into the brush phase, the protein binding properties of these membranes were found to be very resilient upon removal of the system from water, rigorous drying, and rehydration. These results indicate that supported phospholipid bilayers containing lipopolymers show promise as rugged sensor platforms for ligand receptor binding. PMID- 16042483 TI - Interaction of functional dendrimers with multilamellar liposomes: design of a model system for studying drug delivery. AB - Multilamellar liposomes consisting of phosphatidylcholine-cholesterol-dihexadecyl phosphate (19:9.5:1 molar ratio) and dispersed in aqueous or phosphate buffer solutions were interacted with poly(propylene imine) dendrimers which were partially functionalized with guanidinium groups. The remaining toxic external primary amino groups of the dendrimers were reacted with propylene oxide, affording the corresponding hydroxylated derivatives. Microscopic, zeta potential, and dynamic light scattering techniques have shown that liposomal dendrimeric molecular recognition occurs due to the interaction between the complementary phosphate and guanidinium groups. Calcein liposomal entrapment experiments demonstrate a limited leakage, i.e., less than 13%, following liposomes interaction with the modified dendrimers. Calorimetric studies indicate that the enthalpy of the interaction is dependent on the number of guanidinium groups present at the dendrimeric surface and the medium. The process is reversible, and redispersion of the aggregates occurs by adding concentrated phosphate buffer. Two corticosteroid drugs, i.e., betamethasone dipropionate and betamethasone valerate, were encapsulated into the functionalized dendrimers. Drug transport from guanidinylated dendrimers to multilamellar liposomes ranges from 40% to 85%, and it is also dependent on the medium and the degree of dendrimer guanidinylation. PMID- 16042484 TI - Residence time, loading force, pH, and ionic strength affect adhesion forces between colloids and biopolymer-coated surfaces. AB - Exopolymers are thought to influence bacterial adhesion to surfaces, but the time dependent nature of molecular-scale interactions of biopolymers with a surface are poorly understood. In this study, the adhesion forces between two proteins and a polysaccharide [Bovine serum albumin (BSA), lysozyme, or dextran] and colloids (uncoated or BSA-coated carboxylated latex microspheres) were analyzed using colloid probe atomic force microscopy (AFM). Increasing the residence time of an uncoated or BSA-coated microsphere on a surface consistently increased the adhesion force measured during retraction of the colloid from the surface, demonstrating the important contribution of polymer rearrangement to increased adhesion force. Increasing the force applied on the colloid (loading force) also increased the adhesion force. For example, at a lower loading force of approximately 0.6 nN there was little adhesion (less than -0.47 nN) measured between a microsphere and the BSA surface for an exposure time up to 10 s. Increasing the loading force to 5.4 nN increased the adhesion force to -4.1 nN for an uncoated microsphere to a BSA surface and to as much as -7.5 nN for a BSA coated microsphere to a BSA-coated glass surface for a residence time of 10 s. Adhesion forces between colloids and biopolymer surfaces decreased inversely with pH over a pH range of 4.5-10.6, suggesting that hydrogen bonding and a reduction of electrostatic repulsion were dominant mechanisms of adhesion in lower pH solutions. Larger adhesion forces were observed at low (1 mM) versus high ionic strength (100 mM), consistent with previous AFM findings. These results show the importance of polymers for colloid adhesion to surfaces by demonstrating that adhesion forces increase with applied force and detention time, and that changes in the adhesion forces reflect changes in solution chemistry. PMID- 16042485 TI - Self-assembly of a metallosupramolecular coordination polyelectrolyte in the pores of SBA-15 and MCM-41 silica. AB - It is shown that intrinsically stiff chain aggregates of a metallosupramolecular coordination polyelectrolyte (MEPE) can form in the cylindrical nanopores of MCM 41 and SBA-15 silica by self-assembly of its constituents (metal ions and organic ligand). The UV/vis spectra of the resulting MEPE-silica composites exhibit the characteristic metal-to-ligand charge transfer band of the MEPE complex in solution. For the MEPE-silica composite in SBA-15 an iron content of 1.2 wt % was found, corresponding to ca. 10 MEPE chains disposed side by side in the 8 nm wide pores of the SBA-15 matrix. In the case of MCM-41 (pore width < 3 nm), where only one MEPE chain per pore can be accommodated, an iron content of 0.3 wt % was obtained, corresponding to half-filling of the pores. It was also found that MEPE chains spontaneously enter the pores of SBA-15, when a solution of MEPE is exposed to the silica matrix. PMID- 16042486 TI - Ellipsometric and neutron diffraction study of pentane physisorbed on graphite. AB - High-resolution ellipsometry and neutron diffraction measurements have been used to investigate the structure, growth, and wetting behavior of fluid pentane (n C(5)H(12)) films adsorbed on graphite substrates. We present isotherms of the thickness of pentane films adsorbed on the basal-plane surfaces of a pyrolytic graphite substrate as a function of the vapor pressure. These isotherms are measured ellipsometrically for temperatures between 130 and 190 K. We also describe neutron diffraction measurements in the temperature range 11-140 K on a deuterated pentane (n-C(5)D(12)) monolayer adsorbed on an exfoliated graphite substrate. Below a temperature of 99 K, the diffraction patterns are consistent with a rectangular centered structure. Above the pentane triple point at 143.5 K, the ellipsometric measurements indicate layer-by-layer adsorption of at least seven fluid pentane layers, each having the same optical thickness. Analysis of the neutron diffraction pattern of a pentane monolayer at a temperature of 130 K is consistent with small clusters having a rectangular-centered structure and an area per molecule of approximately 37 A(2) in coexistence with a fluid monolayer phase. Assuming values of the polarizability tensor from the literature and that the monolayer fluid has the same areal density as that inferred for the coexisting clusters, we calculate an optical thickness of the fluid pentane layers in reasonable agreement with that measured by ellipsometry. We discuss how these results support the previously proposed "footprint reduction" mechanism of alkane monolayer melting. In the hypercritical regime, we show that the layering behavior is consistent with the two-dimensional Ising model and determine the critical temperatures for layers n = 2-5. PMID- 16042487 TI - Interlayer esterification of layered silicic acid-alcohol nanostructured materials derived from alkoxytrichlorosilane. AB - Layered silicic acid-organic nanohybrid materials consisting of long-chain alkoxy groups attached to thin silica layers have been prepared via esterification of a layered silicic acid-alcohol nanostructured material derived from hexadecoxytrichlorosilane (C(16)H(33)OSiCl(3)). The esterification reaction was performed by heating the layered composite. The detailed characterization of the product heated at 80 degrees C revealed that the interlayer alcohol molecules partly ( approximately 50%) reacted with the interlayer surface silanol groups to form alkoxy groups. Unreacted alcohol molecules were removed by tetrahydrofuran (THF) treatment to form a novel alkoxylated layered silica material. This product retains its structure up to 120 degrees C and has a higher stability in organic solvents if compared with the layered silicic acid-alcohol nanocomposite before esterification, whose structure collapsed over 100 degrees C. Furthermore, various alcohols can be adsorbed into the esterified nanohybrid with the expansion of the interlayer spacing. PMID- 16042488 TI - Colloidal crystallization and transport in stripes and mazes. AB - We study guided crystallization and transport of paramagnetic spheres on top of a magnetic film that arranges its domains into stripes or mazes. In the absence of liquid flow, the paramagnetic spheres are confined within the magnetic domains, and it is shown how the particles self-assemble into several interesting phases depending on the complexity of the domain patterns. We also find that colloids guided through a complex maze exhibit structured patterns that can be controlled by an external magnetic field. The results presented here could help us understand both static and dynamic properties of pattern formation in confined geometries of tunable complexity. PMID- 16042489 TI - Manipulation of the magnetic properties of magnetite-silica nanocomposite materials by controlled Stober synthesis. AB - The paper describes the synthesis and characterization of the magnetic properties of magnetite/silica nanocomposites using a modified Stober method. Magnetite nanoparticles averaging 8-10 nm in diameter and stabilized with oleic acid in toluene were used as the magnetic component of the nanocomposites. SQUID magnetic measurements and ferromagnetic resonance spectroscopy measurements were performed at each stage of the synthesis to understand the properties of the formed composites. Changes of blocking temperature in ZFC/FC SQUID curves correlated with corresponding changes of the resonance field in the ferromagnetic spectra of the sample at each stage of formation. The paper concludes that it is possible to manipulate the magnetic properties of silica/magnetite composite materials by controlling their surface properties and silica coating thickness. PMID- 16042490 TI - Inside-out disruption of silica/gold core-shell nanoparticles by pulsed laser irradiation. AB - Near-infrared (NIR) femtosecond laser irradiation of metallodielectric core-shell silica-gold (SiO(2)-Au) nanoparticles can induce extreme local heating prior to the rapid dissipation of energy caused by the large surface area/volume ratio of nanometer-scale objects. At low pulse intensities, the dielectric silica core is removed, leaving an incomplete gold shell behind. The gold shells with water inside and out still efficiently absorb NIR light from subsequent pulses, showing that a complete shell is not necessary for absorption. At higher pulse intensities, the gold shell itself is melted and disrupted, leading to smaller, approximately 20-nm gold nanoparticles. Spectroscopic measurements show that this disruption is accompanied by optical hole burning of the peak at 730 nm and formation of a new peak at 530 nm. The silica removal and gold shell disruption confirms significant temperature rise of the core-shall nanoparticle. However, the entire process leads to minimal heating of the bulk solution due to the low net energy input. PMID- 16042491 TI - Disk morphology and disk-to-cylinder tunability of poly(acrylic acid)-b poly(methyl acrylate)-b-polystyrene triblock copolymer solution-state assemblies. AB - Disk and cylindrical micellar assemblies were formed through self-organization of poly(acrylic acid)-b-poly(methyl acrylate)-b-polystyrene (PAA-b-PMA-b-PS) amphiphilic triblock copolymers with organic diamines as counterions in water/ tetrahydrofuran (THF) solvent mixtures. The system was investigated by means of transmission electron microscopy and cryogenic transmission electron microscopy. It was found that the assembled-state morphologies could be modified by alteration of the type and concentration of cationic diamine counterion undergoing interaction with the negatively charged, polyelectrolyte PAA corona block, the relative amount of water in the water/THF mixture, and the hydrophobic block chain length. Multivalency of the organic amine counterion was critical for disk formation. It was further demonstrated that a single block copolymer underwent disc-to-cylindrical micellar transitions reversibly with variation in the relative water/THF ratio. The ability to form disks beginning from either THF rich or water-rich solutions indicated that the disk morphology was thermodynamically stable and that THF was important in keeping the micellar structure from becoming kinetically frozen. The nanoassemblies were produced having low size dispersities and were stable for at least one month. Intermediate structures between disks and cylinders were also observed, indicating two distinct kinetic pathways between the two micelle structures. PMID- 16042492 TI - Preparation of highly stable organic steps with a fullerene-based molecule. AB - We report the formation of highly stable Langmuir-Blodgett (LB) organic steps made with a hexa-adduct fullerene-based amphiphile. This amphiphile forms films of excellent quality, with a very low roughness, that are structurally stable: X ray reflectivity spectra recorded on fresh and 12-month-old samples are undiscernible. Such a behavior contrasts with that of more traditional amphiphiles, which are unfortunately well-known for their instability in time. The stability of the films stems, among others, from the spheroidal shape of the constitutive molecules. These experiments show that it is possible to circumvent the major drawback of LB films and to prepare materials more suited for applications. We show that the LB film prepared with this fullerene derivative can successfully be used as thickness gauges for atomic force microscopy or light microscopy studies. PMID- 16042493 TI - Interaction and adhesion properties of polyelectrolyte multilayers. AB - The growth, morphology, and interaction/adhesion properties of supported poly(sodium 4-styrenesulfonate)/poly(allylamine hydrochloride) (PSS/PAH) and DNA/PAH multilayers were investigated by means of surface plasmon resonance spectroscopy, atomic force microscope (AFM) imaging, and AFM-related force measurements. Multilayers were assembled on a prelayer of poly(ethylenimine) (PEI) both with and without drying. SPR results showed a linear growth of the assembly in the case of PSS/PAH multilayers and nonlinear growth for DNA/PAH multilayers. Measurements of forces acting between a bare glass sphere and a multilayer-coated surface indicated repulsive or attractive forces, depending on surface charge, which suggests that, on approach, electrostatic forces dominate. On separation, we observed large pull-off forces in the case of positively charged multilayers and weak pull-off forces in the case negatively charged multilayers. Multiple adhesions and plateau regions observed on separation were interpreted in terms of a bridging of multiple polymer chains between the glass particle and the multilayer and a stretching of the polyelectrolyte loops. The dependence of the pull-off force on the number of deposited layers shows regular oscillations. PMID- 16042494 TI - Poly(oxyethylene) based surface coatings for poly(dimethylsiloxane) microchannels. AB - Control of surface properties in microfluidic systems is an indispensable prerequisite for successful bioanalytical applications. Poly(dimethylsiloxane) (PDMS) microfluidic devices are hampered from unwanted adsorption of biomolecules and lack of methods to control electroosmotic flow (EOF). In this paper, we propose different strategies to coat PDMS surfaces with poly(oxyethylene) (POE) molecules of varying chain lengths. The native PDMS surface is pretreated by exposure to UV irradiation or to an oxygen plasma, and the covalent linkage of POE-silanes as well as physical adsorption of a triblock-copolymer (F108) are studied. Contact angle measurements and atomic force microscopy (AFM) imaging revealed homogeneous attachment of POE-silanes and F108 to the PDMS surfaces. In the case of F108, different adsorption mechanisms to hydrophilic and hydrophobic PDMS are discussed. Determination of the electroosmotic mobilities of these coatings in PDMS microchannels prove their use for electrokinetic applications in which EOF reduction is inevitable and protein adsorption has to be suppressed. PMID- 16042495 TI - Dipping versus spraying: exploring the deposition conditions for speeding up layer-by-layer assembly. AB - Polyelectrolyte film fabrication by successive spraying of polycation and polyanion solutions is described and compared to classic dipping. The poly(styrenesulfonate)/poly(allylamine) system is examined in detail. The influence of various parameters such as spraying time, polyelectrolyte concentration, and effect of film drying during multilayer construction is investigated. It is found that film deposition by spraying is easily controlled and very reliable. The thickness of the multilayers grows linearly with the number of deposition cycles similarly to what is observed when dipping substrates or when polyelectrolyte solutions flow over a surface. The assembly of films is very fast and leads to films with small surface roughness as estimated by atomic force microscopy and X-ray reflectometry. Spray deposition allows achieving regular multilayer growth even under conditions for which dipping fails to produce homogeneous films (e.g., extremely short contact times). Moreover, because drainage constantly removes a certain quantity of the excess material arriving at the surface, one can even skip the rinsing step and, thus, speed up even further the whole buildup process. PMID- 16042496 TI - Packing of simulated friction modifier additives under confinement. AB - Molecular dynamics simulations have been conducted to obtain detailed information regarding molecular structure and packing of surfactant-like "friction modifier" (FM) chains adsorbed to two confining surfaces under sliding conditions. The simulations are interpreted via the density profile, position-dependent fluctuations in the density profile, and parallel, interlayer position correlation functions. Heterogeneous FM structures were obtained. The adsorbed FM chains were found to form semi-ordered monolayers perpendicular to the atomically well-defined surface, with distinct segment positions in the density profile. More fluid layers with broader density profiles and larger fluctuations were formed by some unoriented FM chains oriented preferentially parallel to the surface, between the opposing monolayers. Segment position correlations parallel to the surface in the adsorbed layers were found at separations up to 30 A. These packings persisted for longer than 4 ns under sliding conditions that ranged from 0 to 7.5 m s(-1) (0 to 0.075 A ps(-1)). Monolayer structures were observed to remain unchanged under a wide range of normal pressures, with the corresponding changes in volume occurring in the gap between monolayers or between a monolayer and trapped fluid. PMID- 16042497 TI - Parametric studies of interaction strengths in polymer/CO2 systems: discontinuous molecular dynamics simulations. AB - Discontinuous molecular dynamics simulations are performed on homopolymer/solvent and surfactant/solvent systems. The homopolymer and surfactant molecules are modeled as freely jointed square-well chains. Solvent molecules are modeled as both hard spheres and square-well spheres. We explore how the various interaction parameters affect the types of phase behavior and micellization observed in the homopolymer/solvent and surfactant/solvent systems. Increasing the packing fraction of homopolymers in both hard-sphere solvents and square-well solvents increases the solvent's ability to dissolve homopolymers only when the segment solvent interaction strength exceeds a critical value. Although only upper critical solution temperature (UCST) behavior is observed for homopolymers in hard-sphere solvents, both UCST and lower critical solution temperature (LCST) behavior are observed for homopolymers in square-well solvents, depending upon the interaction strengths and chain length. This indicates that it is necessary to account for the solvent-solvent attraction to model LCST behavior in supercritical CO2. Our simulation results on surfactants in hard-sphere solvents show that it is necessary to account for the interactions experienced by both the head and tail blocks in order to capture the essential features of surfactant/supercritical CO2 systems. PMID- 16042498 TI - Electrophoresis of a charge-regulated sphere normal to a large disk. AB - The electrophoresis of a rigid, charge-regulated, spherical particle normal to a large disk is investigated under the conditions of low surface potential and weak applied electric field. We show that, although the presence of a charged disk does not generate an electroosmotic flow, it affects particle motion appreciably through inducing charge on its surface and establishing an osmotic pressure field. The competition between the hydrodynamic force and the electric force may yields a local extremum in mobility; it is also possible that the direction of particle movement is reversed. In general, if a particle remains at constant surface potential, a decrease in the thickness of double layer has the effect of increasing the electrostatic force acting on it so that its mobility increases. However, this might not be the case for a charged-regulated particle because an excess hydrodynamic force is enhanced. For a fixed separation distance, the influence of a charged disk on mobility may reduce to a minimum if the bulk concentration of hydrogen ion is equal to the dissociation constant of the monoprotic acidic functional groups on particle surface. PMID- 16042499 TI - Joule heating induced transient temperature field and its effects on electroosmosis in a microcapillary packed with microspheres. AB - The Joule heating induced transient temperature field and its effect on the electroosmotic flow in a capillary packed with microspheres is analyzed numerically using the control-volume-based finite difference method. The model incorporates the coupled momentum equation for the electroosmotic velocity, the energy equations for the Joule heating induced temperature distributions in both the packed column and the capillary wall, and the mass and electric current continuity equations. The temperature-dependent physical properties of the electrolyte solution are taken into consideration. The characteristics of the Joule heating induced transient development of temperature and electroosmotic flow fields are studied. Specifically, the simulation shows that the presence of Joule heating causes a noticeable axial temperature gradient in the thermal entrance region and elevates a significant temperature increment inside the microcapillary. The temperature changes in turn greatly affect the electroosmotic velocity by means of the temperature-dependent fluid viscosity, dielectric constant, and local electric field strength. Furthermore, the model predicts an induced pressure gradient to counterbalance the axial variation of the electroosmotic velocity so as to maintain the fluid mass continuity. In addition, under specific conditions, the present model is validated by comparing with the existing analytical model and experimental data from the literature. PMID- 16042500 TI - First-principles analyses and predictions on the reactivity of barrier layers of Ta and TaN toward organometallic precursors for deposition of copper films. AB - We present theoretical studies based on first-principles density functional theory calculations on the mechanisms of chemical vapor deposition of Cu hexafluoracetylacetonato-trimethylvinylsilane (Cu(hfac)(tmvs)) on tantalum surfaces. This process has been used in the past to grow copper films via a disproportionation reaction and was found to exhibit adhesion problems. We show that the Ta surfaces are highly reactive and that the organic ligands in a copper precursor would undergo spontaneous decomposition upon contact with the Ta substrates. This may lead to contamination of the metal surfaces caused by the formation of carbide, fluoride, oxide species, or other fragments of the copper precursor on the barrier layer. We propose a practical solution for these adhesion problems caused by the CVD process by passivating the metal surfaces with N(2) to reduce their activity toward the precursor. Our extensive first principles molecular dynamics simulations under typical deposition conditions predict that, for properly passivated TaN surfaces, only the copper atoms are firmly adsorbed on the surface, with loose Cu-ligand bonds. The ligands are sufficiently stable on these passivated surfaces, remaining slightly above the surface due to the repulsion between the electron-rich N-layer and the electron rich ligand groups, and subsequently liberated upon the disproportionation reaction. PMID- 16042501 TI - The potential adverse health effects of dental amalgam. AB - There is significant public concern about the potential health effects of exposure to mercury vapour (Hg(0)) released from dental amalgam restorations. The purpose of this article is to provide information about the toxicokinetics of Hg(0), evaluate the findings from the recent scientific and medical literature, and identify research gaps that when filled may definitively support or refute the hypothesis that dental amalgam causes adverse health effects. Dental amalgam is a widely used restorative dental material that was introduced over 150 years ago. Most standard dental amalgam formulations contain approximately 50% elemental mercury. Experimental evidence consistently demonstrates that Hg(0) is released from dental amalgam restorations and is absorbed by the human body. Numerous studies report positive correlations between the number of dental amalgam restorations or surfaces and urine mercury concentrations in non occupationally exposed individuals. Although of public concern, it is currently unclear what adverse health effects are caused by the levels of Hg(0) released from this restoration material. Historically, studies of occupationally exposed individuals have provided consistent information about the relationship between exposure to Hg(0) and adverse effects reflecting both nervous system and renal dysfunction. Workers are usually exposed to substantially higher Hg(0) levels than individuals with dental amalgam restorations and are typically exposed 8 hours per day for 20-30 years, whereas persons with dental amalgam restorations are exposed 24 hours per day over some portion of a lifetime. This review has uncovered no convincing evidence pointing to any adverse health effects that are attributable to dental amalgam restorations besides hypersensitivity in some individuals. PMID- 16042502 TI - Toxicological aspects of the South American herbs cat's claw (Uncaria tomentosa) and Maca (Lepidium meyenii) : a critical synopsis. AB - Recent exceptional growth in human exposure to natural products known to originate from traditional medicine has lead to a resurgence of scientific interest in their biological effects. As a strategy for improvement of the assessment of their pharmacological and toxicological profile, scientific evidence-based approaches are being employed to appropriately evaluate composition, quality, potential medicinal activity and safety of these natural products. Using this approach, we comprehensively reviewed existing scientific evidence for known composition, medicinal uses (past and present), and documented biological effects with emphasis on clinical pharmacology and toxicology of two commonly used medicinal plants from South America with substantial human exposure from historical and current global use: Uncaria tomentosa (common name: cat's claw, and Spanish: una de gato), and Lepidium meyenii (common name: maca). Despite the geographic sourcing from remote regions of the tropical Amazon and high altitude Andean mountains, cat's claw and maca are widely available commercially in industrialised countries. Analytical characterisations of their active constituents have identified a variety of classes of compounds of toxicological, pharmacological and even nutritional interest including oxindole and indole alkaloids, flavonoids, glucosinolates, sterols, polyunsaturated fatty acids, carbolines and other compounds. The oxindole alkaloids from the root bark of cat's claw are thought to invoke its most widely sought-after medicinal effects as a herbal remedy against inflammation. We find the scientific evidence supporting this claim is not conclusive and although there exists a base of information addressing this medicinal use, it is limited in scope with some evidence accumulated from in vitro studies towards understanding possible mechanisms of action by specific oxindole alkaloids through inhibition of nuclear factor (NF)-kappaB activation. Although controlled clinical studies have demonstrated reduction in pain associated with cat's claw intake in patients with various chronic inflammatory disorders, there is insufficient clinical data overall to draw a firm conclusion for its anti-inflammatory effects. An important observation was that experimental results were often dependent upon the nature of the preparation used. It appears that the presence of unknown substances has an important role in the overall effects of cat's claw extracts is an important factor for consideration. The available animal toxicological studies did not indicate severe toxicity from oral intake of cat's claw preparations but rather were suggestive of a low potential for acute and subacute oral toxicity, and a lack of evidence to demonstrate genotoxic potential and mutagenic activity. Maca is a clear example of a herb with substantial medicinal use in traditional herbal medicine by indigenous cultures in South America since the first recorded knowledge of it in the seventeenth century. The hypocotyls of maca are the edible part of the plant used for nutritional and proposed fertility-enhancing properties. Maca has been described to possess many other medicinal properties in traditional herbal medicine but only a few of them have been well studied scientifically. Published clinical studies of maca seem to be related to its property as a nutrient, for male fertility and for energy. There are inadequate data regarding the precise mechanism of action of maca. Some studies suggest that secondary metabolites found in maca extracts are important constituents responsible for its physiological effects. Maca has been reported in the scientific literature to have a low degree of acute oral toxicity in animals and low cellular toxicity in vitro. An important finding unveiled by this review is the importance of standardisation in quality and additional basic and clinical research to scientifically validate and understand composition, biological activity, safety and risk. Development of a comprehensive pharmacological and toxicological profile through critical evaluation of existing and future experimental data, especially carefully conducted clinical studies would facilitate the scientific evidence-based approach to understanding potential biological effects of these major traditionally based herbals in current global use. PMID- 16042503 TI - Organophosphate-induced delayed polyneuropathy. AB - Organophosphate-induced delayed polyneuropathy (OPIDP) is a rare toxicity resulting from exposure to certain organophosphorus (OP) esters. It is characterised by distal degeneration of some axons of both the peripheral and central nervous systems occurring 1-4 weeks after single or short-term exposures. Cramping muscle pain in the lower limbs, distal numbness and paraesthesiae occur, followed by progressive weakness, depression of deep tendon reflexes in the lower limbs and, in severe cases, in the upper limbs. Signs include high-stepping gait associated with bilateral foot drop and, in severe cases, quadriplegia with foot and wrist drop as well as pyramidal signs. In time, there might be significant recovery of the peripheral nerve function but, depending on the degree of pyramidal involvement, spastic ataxia may be a permanent outcome of severe OPIDP. Human and experimental data indicate that recovery is usually complete in the young. At onset, the electrophysiological changes include reduced amplitude of the compound muscle potential, increased distal latencies and normal or slightly reduced nerve conduction velocities. The progression of the disease, usually over a few days, may lead to non-excitability of the nerve with electromyographical signs of denervation. Nerve biopsies have been performed in a few cases and showed axonal degeneration with secondary demyelination. Neuropathy target esterase (NTE) is thought to be the target of OPIDP initiation. The ratio of inhibitory powers for acetylcholinesterase and NTE represents the crucial guideline for the aetiological attribution of OP-induced peripheral neuropathy. In fact, pre-marketing toxicity testing in animals selects OP insecticides with cholinergic toxicity potential much higher than that to result in OPIDP. Therefore, OPIDP may develop only after very large exposures to insecticides, causing severe cholinergic toxicity. However, this was not the case with certain triaryl phosphates that were not used as insecticides but as hydraulic fluids, lubricants and plasticisers and do not result in cholinergic toxicity. Several thousand cases of OPIDP as a result of exposure to tri-ortho-cresyl phosphate have been reported, whereas the number of cases of OPIDP as a result of OP insecticide poisoning is much lower. In this article, we mainly discuss OP pesticide poisoning, particularly when caused by chlorpyrifos, dichlorvos, isofenphos, methamidophos, mipafox, trichlorfon, trichlornat, phosphamidon/mevinphos and by certain carbamates. We also discuss case reports where neuropathies were not convincingly attributed to fenthion, malathion, omethoate/dimethoate, parathion and merphos. Finally, several observational studies on long-term, low-level exposures to OPs that sometimes reported mild, inconsistent and unexplained changes of unclear significance in peripheral nerves are briefly discussed. PMID- 16042504 TI - Interpretation of analytical toxicology results in life and at postmortem. AB - Interpretation of analytical toxicology results from live patients is sometimes difficult. Possible factors may be related to: (i) the nature of the poison(s) present; (ii) sample collection, transport and storage; (iii) the analytical methodology used; (iv) the circumstances of exposure; (v) mechanical factors such as trauma or inhalation of stomach contents; and (vi) pharmacological factors such as tolerance or synergy. In some circumstances, detection of a drug or other poison may suffice to prove exposure. At the other extreme, the interpretation of individual measurements may be simplified by regulation. Examples here include whole blood alcohol (ethanol) in regard to driving a motor vehicle and blood lead assays performed to assess occupational exposure. With pharmaceuticals, the plasma or serum concentrations of drugs and metabolites attained during treatment often provide a basis for the interpretation of quantitative measurements. With illicit drugs, comparative information from casework may be all that is available. Postmortem toxicology is an especially complex area since changes in the composition of fluids such as blood depending on the site of collection from the body and the time elapsed since death, amongst other factors, may influence the result obtained. This review presents information to assist in the interpretation of analytical results, especially regarding postmortem toxicology. Collection and analysis of not only peripheral blood, but also other fluids/tissues is usually important in postmortem work. Alcohol, for example, can be either lost from, or produced in, blood especially if there has been significant trauma, hence measurements in urine or vitreous humour are needed to confirm the reliability of a blood result. Measurement of metabolites may also be valuable in individual cases. PMID- 16042505 TI - Analytical toxicology: guidelines for sample collection postmortem. AB - The reliability and relevance of any analytical toxicology result is determined in the first instance by the nature and integrity of the specimen(s) submitted for analysis. This article provides guidelines for sample collection, labelling, transport and storage, especially regarding specimens obtained during a postmortem examination. Blood (5 mL) should be taken from two distinct peripheral sites, preferably left and right femoral veins, taking care not to draw blood from more central vessels. Urine (if available), vitreous humour (separate samples from each eye), a representative portion of stomach contents, and liver (10-20 g, right lobe) are amongst other important specimens. A preservative (sodium fluoride, 0.5-2% weight by volume (w/v) should be added to a portion of the blood sample/the sample from one vein, and to urine. Leave a small (10-20% headspace) in tubes containing liquids if they are likely to be frozen. Precautions to minimise the possibility of cross-contamination of biological specimens must be taken, especially if volatile poison(s) may be involved. If death occurred in hospital, any residual antemortem samples should be sought as a matter of urgency. Hair/nail collection should be considered if chronic exposure is suspected, for example, in deaths possibly related to drug abuse. A lock of hair the width of a pen tied at the root end is required for a comprehensive drug screen. The value of providing as full a clinical/occupational/circumstantial history as possible together with a copy of the postmortem report (when available) and of implementing chain-of-custody procedures when submitting samples for analysis cannot be over-emphasised. PMID- 16042506 TI - Bimolecular hydrogen abstraction from phenols by aromatic ketone triplets. AB - Absolute rate constants for hydrogen abstraction from 4-methylphenol (para cresol) by the lowest triplet states of 24 aromatic ketones have been determined in acetonitrile solution at 23 degrees C, and the results combined with previously reported data for roughly a dozen other compounds under identical conditions. The ketones studied include various ring-substituted benzophenones and acetophenones, alpha,alpha,alpha-trifluoroacetophenone and its 4-methoxy analog, 2-benzoylthiophene, 2-acetonaphthone, and various other polycyclic aromatic ketones such as fluorenone, xanthone and thioxanthone, and encompass n,pi*, pi,pi*(CT) and arenoid pi,pi* lowest triplets with (triplet) reduction potentials (E(red)*) varying from about -10 to -38 kcal mol(-1). The 4 methylphenoxyl radical is observed as the product of triplet quenching in almost every case, along with the corresponding hemipinacol radical in most instances. Hammett plots for the acetophenones and benzophenones are quite different, but plots of log k(Q) vs E(red)* reveal a common behavior for most of the compounds studied. The results are consistent with reaction via two mechanisms: a simple electron-transfer mechanism, which applies to the n,pi* triplet ketones and those pi,pi* triplets that possess particularly low reduction potentials, and a coupled electron-/proton-transfer mechanism involving the intermediacy of a hydrogen bonded exciplex, which applies to the pi,pi* ketone triplets. Ketones with lowest charge-transfer pi,pi* states exhibit rate constants that vary only slightly with triplet reduction potential over the full range investigated; this is due to the compensating effect of substituents on triplet state basicity and reduction potential, which both play a role in quenching by the hydrogen-bonded exciplex mechanism. Ketones with arenoid pi,pi* states exhibit the fall-off in rate constant that is typical of photoinduced electron transfer reactions, but it occurs at a much higher potential than would be normally expected due to the effects of hydrogen-bonding on the rate of electron-transfer within the exciplex. PMID- 16042507 TI - Human dignity as a criterion for science policy. PMID- 16042513 TI - Self-assessment in medical practice: a further concern about the conventional research paradigm. PMID- 16042514 TI - Simulation-based training of internal medicine residents in advanced cardiac life support protocols: a randomized trial. AB - BACKGROUND: Internal medicine residents must be competent in Advanced Cardiac Life Support (ACLS) for board certification. PURPOSE: The purpose was to use a medical simulator to assess baseline proficiency in ACLS and determine the impact of an intervention on skill development. METHOD: This was a randomized trial with wait-list controls. After baseline evaluation in all residents, the intervention group received 4 education sessions using a medical simulator. All residents were then retested. After crossover, the wait-list group received the intervention, and residents were tested again. Performance was assessed by comparison to American Heart Association guidelines for treatment of ACLS conditions with interrater and internal consistency reliability estimates. RESULTS: Performance improved significantly after simulator training. No improvement was detected as a function of clinical experience alone. The educational program was rated highly. PMID- 16042515 TI - Reliability and validity of checklists and global ratings by standardized students, trained raters, and faculty raters in an objective structured teaching environment. AB - BACKGROUND: Objective structured teaching exercises (OSTEs) are relatively new in medical education, with few studies that have reported reliability and validity. PURPOSE: To systematically examine the impact of OSTE design decisions, including number of cases, choice of raters, and type of scoring systems used. METHODS: We examined the impact of number of cases and raters using generalizability theory. We also compared scores from standardized students (SS), faculty raters (FR) and trained graduate student raters (TR), and examined the relation between behavior checklist ratings and global perception scores. RESULTS: Generalizability (g) coefficients for checklist scores were higher for SSs than TRs. The g estimates based on SSs' global scores were higher than g estimates for FRs. SSs' checklist scores were higher than TRs' checklist scores, and SSs' global evaluations were higher than FRs' and TRs' global scores. TRs' relative to SSs' global perceptions correlated more highly with checklist scores. CONCLUSIONS: SSs provide more generalizable checklist scores than TRs. Generalizability estimates for global scores from SSs and FRs were comparable. SSs are lenient raters compared to TRs and FRs. PMID- 16042516 TI - Expectations of groups versus pairs of attendings and residents about phone communications and bedside evaluation of hospitalized patients. AB - BACKGROUND: Residents covering attendings' patients must decide when to call the attending and when to evaluate patients at the bedside for new clinical events. Conflicts arise when residents and attendings disagree about these decisions. PURPOSE: Our purpose was to determine differences in expectations between attendings and residents concerning the need for phone communication about, and bedside evaluation of, new patient problems in hospitalized patients. METHODS: Teaching attendings and categorical medical residents rated a series of 15 clinical vignettes regarding the importance of phone communication about the problems depicted in the vignettes and rated a second series of 9 vignettes about the need for bedside evaluation of problems depicted. RESULTS: Mean Likert scores were similar between residents and attendings for 70% of the scenarios. When analyzed for disagreement within hypothetical resident/attending pairs, the survey revealed that disagreement would be expected to occur as often as 33% of the time for phone communication and up to 43% of the time for bedside evaluation. Attendings disagreed with each other 41% of the time. Residents were less likely to say they would call attendings at night than attendings wanted to be called (odds ratio=0.74, p=.04). CONCLUSION: There was substantial disagreement among hypothetical resident--attending pairs regarding need for phone communication and bedside evaluation. Residents and attendings need to discuss their expectations regarding these issues to avoid conflict. PMID- 16042517 TI - Using standardized patient outcome to measure the effect of teaching asthma related patient education and information-giving skills to medical students. AB - BACKGROUND: Patient education and giving information is a core skill that improves patient adherence and medical outcomes. PURPOSE: To evaluate the impact of a teaching intervention on 3rd-year students' competency in patient education and information giving about asthma medication delivery. METHODS: Students (n=81) completed a 1-hr teaching intervention of didactics followed by role playing of asthma patient education scenarios. Using a standardized patient post intervention, patient education and information-giving skills about spacer/metered dose inhalers were scored overall and on a 12-item checklist and compared to a control group (n=70). Students' knowledge was evaluated using a short answer test. RESULTS: The performance of intervention students on overall patient education, 10 of the 12 checklist items, and the test was significantly higher than controls but did not approach competency. CONCLUSIONS: The 1-hr intervention improved clinical performance and knowledge, but students did not become competent. Future studies should investigate how competence in this and other core patient education skills can be successfully achieved. PMID- 16042518 TI - Writing-skills development in the health professions. AB - BACKGROUND: Studies have found that students in the medical professions often lack the writing skills required during their education and career. One contributing factor to this deficiency is that writing tends to be discipline specific, rather than requiring general skills acquired in undergraduate schools. PURPOSE: To determine the extent to which a rigorous writing exercise impacted the quality of students' medical writing based on a specified rubric. METHOD: In the context of a basic science course, we developed 6 weekly writing exercises called Question of the Week, along with a rubric for scoring students' work. The rubric evaluated 6 specific aspects of students' writing including Comprehensiveness/Thoroughness, Accuracy, Conciseness, Logical Organization, Justification of Assertions, and Use of Appropriate Terminology. RESULTS: Except for Justification of Assertions and Accuracy, which did not change, scores for all categories improved between Weeks 1 and 2. Use of Appropriate Terminology was the only category for which scores increased after Week 2. CONCLUSION: The clearest indication of writing development came from students' augmented ability to use medical terminology in appropriate ways. This is an important observation, given that each Question of the Week covered a separate body system, characterized by distinctly different terms and jargon. We concluded that students need much more practice to attain the level of proficiency outlined by our rubric. PMID- 16042519 TI - The nature of the interaction between participants and facilitators in online asynchronous continuing medical education learning environments. AB - BACKGROUND: MDcme.ca offers an online asynchronous continuing medical education (CME) environment for family physicians. The nature of participation in online CME using computer-mediated conferencing (CMC) discussion systems and the characteristics of interaction between participants and facilitators is not well understood. PURPOSE: To examine the association between participant and facilitator participation in online asynchronous CME learning environments. METHODS: We analyzed registration and participation data including participant and facilitator postings to an asynchronous CMC discussion system for CME programs offered through MDcme.ca using frequency counts. Pearson r correlation was used to assess the association between numbers of participants and participant postings, facilitator postings and participant postings, and the number of discussion items accessed and participant postings. RESULTS: MDcme.ca offered 25 programs to 327 registrants, of whom 180 actually participated by accessing course materials and online discussion postings. Pearson r correlations showed a significant association between the number of participants and the number of postings, the number of facilitator postings and the number of participant postings, and the mean number of discussion items accessed by participants and the overall number of participant postings. DISCUSSION: Both the number of facilitator postings and the number of discussion items accessed by participants appeared to be important determinants of the amount of interaction that will occur in asynchronous online CME. Curriculum planners and facilitators of asynchronous CME need to be aware of the importance of fostering interactive, stimulating discussions if the potential of asynchronous learning for physicians is to reach its potential in supporting higher levels of critical, reflective, practice-based learning. PMID- 16042520 TI - Outcomes of a teaching scholars program to promote leadership in faculty development. AB - BACKGROUND: Provision of adequate opportunities for faculty to receive training in teaching skills requires persons with faculty development expertise to provide this training. PURPOSE: In this study, we examined outcomes of a teaching scholars program (TSP) aimed at promoting leaders in faculty development in teaching skills in individual departments and throughout the institution. METHOD: A 3-year TSP, established in 1999, combines monthly meetings and other activities to train faculty representing departments throughout the College of Medicine to provide faculty development in teaching skills. Analysis of program participants' curriculum vitae was conducted to compare preprogram and postprogram faculty development activities. RESULTS: There was substantial increase in program participants' facilitation of teaching skills workshops and development of teaching improvement systems within their home departments. Participants also facilitated an increased number of college-wide workshops. Secondary outcomes included increases in educational leadership positions and education-related presentations and publications. CONCLUSION: The TSP is a successful local model for increasing the resources available for teaching improvement. PMID- 16042521 TI - Face-to-face clinical skills feedback: lessons from the analysis of standardized patient's work. AB - BACKGROUND: Current evidence suggests that trainees are evaluated less stringently when feedback is given face-to-face, limiting its value. PURPOSE: We expected that standardized patients (SPs) would also be more lenient in scoring when they gave immediate feedback to the students. METHODS: Data from 6 clinical skills assessments of students in Years 1 to 4 of medical school were used. Comparisons in scoring were made for the cases on which students received feedback and those where they did not. Analysis was done using a hierarchical linear regression model to test for significant differences. RESULTS: The results indicated no significant difference in scoring for the history and physical exam checklists. There was a small but significant difference on the measure of interpersonal and interviewing skills. CONCLUSIONS: The SPs were trained to avoid stringency, leniency, and restricted range in scoring students. These training methods may be useful for training faculty to give face-to-face feedback. PMID- 16042522 TI - The family medicine clerkship over the past 10 years at Ben Gurion University of the Negev. AB - BACKGROUND: Most medical teaching takes place in the academic hospital setting and students continue to have little exposure to community-based medicine. DESCRIPTION: The Goldman School of Medicine of the Ben-Gurion University of the Negev is a 6-year program. The 6-week family medicine clerkship takes place in the 5th year. It includes experience with home care, patient and family centered care, ongoing comprehensive responsibility, common office clinical skills, and specific organization of practicing in the community. At the end of the clerkship, students are asked to complete anonymous feedback forms to evaluate the clerkship, its formal curriculum, and the teachers. EVALUATION: Over the last 3 years, 157 general course evaluation forms were returned (84.4% compliance). The mean general evaluation and satisfaction score was 3.4 out of 4, ranking it very high compared to other clerkships in our medical school. In the same time period, 176 specific teachers evaluation forms were returned, with a mean score of 4.6 (2=weak; 5=excellent), compared with a mean score of 4.5 in 1,485 teachers evaluation forms from other clinical clerkships. CONCLUSIONS: As the focus of patient care shifts to the community, the focus of medical teaching should follow the same route. We hope that our experience will help in promoting this change and in developing new ideas for teaching family medicine and primary care. PMID- 16042523 TI - Developing and implementing universal guidelines for oral patient presentation skills. AB - BACKGROUND: Oral case presentations are important for patient care and clinical education. Previously published attempts to improve oral presentation skills have been labor intensive and have focused primarily on medical students. DESCRIPTION: We created a multifaceted intervention to improve oral case presentations of medical students and internal medicine residents. Our intervention included a written handout with detailed instructions, pocket cards, model presentations, and multiple teaching sessions. The intervention was developed by consensus, supported by the department of medicine and endorsed by key faculty within the department. In addition to soliciting feedback from students and residents, we evaluated our intervention with prospectively acquired ratings of student oral case presentation skills recorded on a standardized evaluation for the internal medicine clerkship. EVALUATION: Students, residents, and faculty gave positive feedback, although they noted some practical obstacles to effective oral case presentations. After dissemination of the guidelines, 44% of students (42/96) were rated as "excellent" in oral presentation skills on standardized evaluations of the internal medicine clerkship, compared to 30% in the previous academic year (33/111; odds ratio [OR]=1.8, 95% confidence interval=1.04-3.3; p=.036). CONCLUSION: A multifaceted intervention improved medical student oral case presentation skills. Although participants noted barriers that need further attention, we demonstrated modest improvement in student performance. PMID- 16042524 TI - Synchronous distance interactive classroom conferencing. AB - BACKGROUND: New medical schools have been opened in the eastern and southeastern regions of the country. They are also in great need of basic medical science teachers. However, due to security reasons over the past two decades, teachers from the established universities do not desire to travel to these medical schools for lectures. The objective of this study was to develop a synchronous classroom conferencing system to teach basic science courses between two general purpose technology enhanced classrooms of two different universities--Istanbul University (IU) and Istanbul and Harran University (HU), Urfa--located 1,500 miles apart in Turkey. DESCRIPTION: I videostreamed the instructor, content from document camera, Power Point presentations at IU, and the students at both places, IU and HU. In addition, I synchronously broadcast two whiteboards by attaching two mimio devices to the two blackboards in the IU classroom to capture and convert everything written or drawn on them into broadcasting over the intranet. This technique is called "boardcasting," which allows users to stream ink and audio together over the Internet or intranet live. A total of 260 students at IU and 150 students at HU were involved. Off-campus HU students also have asynchronous access to the stored lecture video materials at any time. Midterm and final examinations were administered simultaneously using the same questions at both sites in two universities under the observation of the teaching faculty using the very same system. EVALUATION: This system permitted interaction between the students in the class at IU and remote-campus students at HU and the instructor in real time. The instructors at IU were able to maintain a significant level of spontaneity in using their multimedia materials and electronic whiteboards. The mean midterm and final exam scores of students at both universities were similar. CONCLUSION: The system developed in this study can be used by the medical faculty at the main teaching hospitals to deliver their lectures in real time to the medical students when they are on placement in general practices, ambulatory clinics, and district hospitals in rural areas or to the students of other medical schools. This is an efficacious system in developing countries where availability of high-speed networks and cost of communication are major concerns. PMID- 16042525 TI - Internists training medical residents in pelvic examination: impact of an educational program. AB - BACKGROUND: Pelvic examination is an important component of the primary care of women by internists, but training beyond medical school is rare. DESCRIPTION: We created an internist-run educational program for 1st-year medical residents in pelvic examination. The program consisted of 4 weekly patient-care sessions with 2 to 3 patients seen by each resident each session. Internists supervised each exam and gave real-time feedback and utilized a skills-assessment checklist during the first and last exams of the program to give comprehensive, formative feedback. EVALUATION: We evaluated the program using a self-assessment questionnaire concerning pelvic examination competency and attitudes, which utilized a 5-point Likert scale and was administered prior to, and 3 months after, the program. A total of 37 participants completed the program and reported improvements in competency and a trend toward an increased likelihood of performing exams. CONCLUSIONS: This program, in which internists trained medical residents in pelvic examination, utilized real patients, improved participants self-assessed competency, and may increase the likelihood of residents performing pelvic examination in primary care. PMID- 16042526 TI - Narrative means to humanistic ends. AB - BACKGROUND: Efforts to "rehumanize" medical education through curriculum reform and program development have been numerous and ongoing in recent years. One particularly intriguing contribution has come from the area of narrative studies. It is now common to use literature in general, and physician--patient narratives in particular, both to expand students' understanding of the clinical encounter and to sensitize them to the humanistic aspects of medicine. DESCRIPTION: In this article, we describe the process by which we have introduced key insights from and elements of narrative theory into our 1st-year clinical skills program. Rather than limiting our efforts to the use of literature and to the description of individual narrative encounters, however, we have framed our entire course as an exercise in narrative construction. We refer to this process as "narrative structuring." EVALUATION: A combination of short essays on topics related to the various literary materials utilized in the course, written reports on ethical aspects of the clinical cases presented in the clinical skills sessions, and student journaling are integral components of the evaluation of this course. CONCLUSION: Characterizing our course in terms of narrative structuring serves both to integrate the various elements of our complex curriculum around a common theme and to remind both students and faculty alike of the privileges and responsibilities we share as we participate in the writing of one another's stories. PMID- 16042527 TI - Comedy workshop: an enjoyable way to develop multiple-choice questions. AB - PURPOSE: To describe an innovative method of developing multiple-choice items for a board certification examination. SUMMARY: The development of appropriate multiple-choice items is definitely more of an art, rather than a science. The comedy workshop format for developing questions for a certification examination is similar to the process used by comedy writers composing scripts for television shows. This group format dramatically diminishes the frustrations faced by an individual question writer attempting to create items. The vast majority of our comedy workshop participants enjoy and prefer the comedy workshop format. It provides an ideal environment in which to teach and blend the talents of inexperienced and experienced question writers. CONCLUSION: This is a descriptive article, in which we suggest an innovative process in the art of creating multiple-choice items for a high-stakes examination. PMID- 16042528 TI - Mini-immersion in medical Spanish for family practice residents. AB - BACKGROUND: Residency programs vary widely in types of training to increase cultural competency and sensitivity. Moreover, few empirical studies exist regarding effectiveness of these experiences. The purpose of our study was to offer and evaluate a linguistic mini-immersion, !Espanol Rapido!, required of 8 new family practice interns during their orientation month at the University of North Carolina. DESCRIPTION: The curriculum was based on accelerative learning, a pedagogy that considers all parts of the brain, the paraconscious, and the role of the emotions. EVALUATION: Pretests and posttests and a postimmersion and 6 month follow-up evaluation indicated that the mini-immersion was successful. A simple t test for paired samples showed a significant improvement in interns' comprehension after the 6-day immersion, t(7)=11.399, p<.000. CONCLUSION: This brief experience should be viewed only as a first step in a long-term plan for a comprehensive curriculum to prepare family practice residents as culturally competent practitioners. PMID- 16042533 TI - Re: a study to determine: should conventional amounts of eye muscle surgery for horizontal binocular deviations be changed when oblique muscle weakening procedures are simultaneously performed? PMID- 16042536 TI - Screening for amblyopia in preverbal children with photoscreening photographs: IV. Interobserver variability in photograph grading: origin and method of reduction. AB - BACKGROUND: Earlier detection of childhood vision disorders is a prominent goal of vision screening. The Medical Technology and Innovation (MTI) PhotoScreener addresses this objective. Use of this camera does not require verbal feedback and may be administered early in a child's development. Decreasing the variability in photograph grading results will boost the utility of any photoscreening system. This report aims to understand and to decrease intra- and inter-observer variability in grading photoscreening photographs. METHODS: We dissected the photograph grading process and quantified the intra- and inter-observer agreement using intraclass plot correlation coefficients. We evaluated the outcome of a two grader verification system vs. adjudicated measurements with Receiver Operator Characteristic (ROC) curves. PARTICIPANTS: Data on 955 children under 5 years of age, normal except for refractive error, each with complete photoscreening and eye examination data, culled from two previous studies. MAIN OUTCOME MEASURES: Intra- and inter-observer agreement in measuring bright crescent dimensions and pupillary diameters. Sensitivity and specificity of detection of hyperopia. RESULTS: Measurements of bright crescents are associated with greater variability than are measurements of pupillary diameters. Recognition and omission of light "rim" measurements from photograph grading will result in superior inter-observer agreement. Photograph independent errors increase variability and may be corrected by remeasurement. A verification system in which the most discrepant 5% of measurements are redone results in ROC curves similar to adjudicated dimension. CONCLUSIONS: We conclude: 1) two novices grading photographs can do as well as one expert in most cases; 2) the proposed grading methodology has undergone statistical validation and can be used in other areas of ophthalmology and medicine; and 3) inter-observer variability, one of the limitations of photoscreening photograph grading, can be reduced. For 95% of the photographs, two novices achieve similar true positive and true negative values with or without adjudication. PMID- 16042537 TI - Bagolini striated glasses test and lesions of the optic chiasm. AB - PURPOSE: To examine the usefulness of the Bagolini Striated Glasses Test to identify patients with a lesion of the optic chiasm. SUBJECTS AND METHODS: Twelve patients who were diagnosed with a lesion of the optic chiasm by MRI were examined with the Bagolini Striated Glasses Test. Discrepancies of the test between monocular and binocular conditions were analyzed. Visual acuity, visual field and stereo tests were also administered and assessed. RESULTS: Nine out of 12 cases (75%) showed conflicting results with the Bagolini Striated Glasses Test between monocular and binocular conditions. With monocular testing, essentially normal stripes were observed in each eye. In binocular testing, stripes which were projected upon nasal retina were suppressed bilaterally (bitemporal hemianopia pattern) ("mountain pattern") or monocularly (monocular temporal hemianopia pattern). CONCLUSION: The Bagolini Striated Glasses Test can be used as a simple differential diagnostic test for lesions of the optic chiasm. The "mountain pattern" seen binocularly on this test may be pathognomonic for such lesions, which may otherwise be asymptomatic and/or overlooked. PMID- 16042538 TI - An unusual case of congenital convergent convergent strabismus fixus. AB - BACKGROUND: Convergent strabismus fixus is classified into congenital and acquired types; however, there are only a few detailed case reports of congenital convergent strabismus fixus. Here, we report a rare case of this condition in which the eyeball was shown by imaging diagnosis to be fixed with a hard cord like material in the orbit. To our knowledge, there have been no previous reports of congenital convergent strabismus fixus caused by an abnormal cord-like material that was clearly identifiable by imaging. CASE REPORT: The patient was a 16 year old female who visited our department for consultation regarding restoration of normal appearance of her strabismus. She had undergone strabismus surgery at the age of 3 years, but a forced duction test was strongly positive in all directions, and the eyeball was not mobile at the time. The initial surgery performed was a tenotomy of the medial rectus muscle and inferior rectus muscle and resection of the lateral rectus muscle of the right eye. The eye position and eye movement remained unchanged after surgery. At this time, MRI of the orbital region was performed, and an abnormal cord-like material connecting the posterior eyeball to the orbital wall was detected in the orbit. The cord-like material was pulling strongly on the posterior eyeball, congenitally fixing the eye position internally. CONCLUSION: We experienced a rare case in which a hard cord-like material in the orbit congenitally fixed the position of the eyeball. On MRI, the intensity of the cord-like material was clearly different from those of the muscle and nerve tissues. Considering the linearity of the cord-like material and the results of a forced duction test, the cord-like material probably consisted of fairly hard tissue, similar to bone. PMID- 16042540 TI - 'Nature-inspired' drug-protein complexes as inhibitors of Abeta aggregation. AB - Protein-protein interactions are a regulatory mechanism for a number of physiological and pathological cellular processes. Neurodegenerative diseases, such as AD (Alzheimer's disease), are associated with the accelerated production or delayed clearance of protein aggregates. Hence, inhibition of pathologic protein-protein interactions is a very attractive mechanism for drug development. This review focuses on a novel therapeutic strategy to inhibit the de novo formation of protein aggregates. Inspired by strategies used in Nature and optimized over millions of years of evolution, we have created a bifunctional molecule [SLF (synthetic ligand for FK506-binding protein)-CR (Congo Red)] that is able to block Abeta (amyloid beta) aggregation by borrowing the surface and steric bulk of a cellular chaperone. PMID- 16042541 TI - The production of hydrogen peroxide during early-stage protein aggregation: a common pathological mechanism in different neurodegenerative diseases? AB - By means of an ESR spin-trapping method, we have shown that Abeta (amyloid beta), alpha-synuclein and various toxic forms of the prion protein all appear to generate H2O2 in vitro. A fundamental molecular mechanism underlying the pathogenesis of cell death in several different neurodegenerative diseases could be the direct production of H2O2 during the early stages of protein aggregation. PMID- 16042542 TI - Molecular chaperones as therapeutic targets in amyotrophic lateral sclerosis. AB - Neurodegenerative diseases are characterized by a number of common hallmarks, such as the presence of intracellular aggregates and activation of the apoptotic cell-death pathway. Intracellular chaperones, responsible for protein integrity and structural repair, may play a crucial role in the progression of a disease. In this paper, we aim to summarize our understanding of the role and potential of a particular family of chaperones, the heat-shock proteins, in neurodegeneration, by focusing our discussion on models of motoneuron death. PMID- 16042543 TI - Disease modifying strategies for the treatment of Alzheimer's disease targeted at modulating levels of the beta-amyloid peptide. AB - AD (Alzheimer's disease) is characterized neuropathologically by the presence of amyloid plaques, neurofibrillary tangles and profound grey matter loss. The 'amyloid' hypothesis postulates that the toxic Abeta (amyloid beta) peptide, enzymatically derived from the proteolytic processing of a larger protein called APP (amyloid precursor protein), is one of the principal causative factors of neuronal cell death in the brains of AD patients. As such, methods for lowering Abeta levels in the brain are of significant interest with regard to identifying novel disease modifying therapies for the treatment of AD. In this review, we will review a variety of approaches and mechanisms capable of modulating levels of Abeta. PMID- 16042544 TI - Presenilin 1: more than just gamma-secretase. AB - Presenilin 1 plays a central catalytic role in the gamma-secretase processing of amyloid precursor protein, Notch and many other substrates. However, this core component clearly mediates independently several other physiological roles in the cell/neuron. Besides its involvement in beta-catenin degradation, we discuss here the recent implication of presenilin 1 in the turnover of the intercellular cell adhesion molecule, telencephalin, through a degradation route that bears autophagic characteristics. Activation of the endosomal/lysosomal system in general and autophagic degradation in particular, is finally briefly discussed in the context of neurodegenerative diseases. PMID- 16042545 TI - Synaptic plasticity disruption by amyloid beta protein: modulation by potential Alzheimer's disease modifying therapies. AB - AD (Alzheimer's disease) is characterized by a progressive and devastating mental decline that is usually presaged by impairment of a form of memory dependent on medial temporal lobe structures, including the hippocampus. The severity of clinical dementia correlates positively with the cerebral load of the AD-related protein Abeta (amyloid beta), particularly in its soluble form rather than the insoluble fibrillar Abeta found in amyloid plaques. Recent research in animal models of AD has pointed to a potentially important role for rapid disruptive effects of soluble species of Abeta on neural function in causing a relatively selective impairment of memory early in the disease. Our experiments assessing the mechanisms of Abeta inhibition of LTP (long-term potentiation), a correlate of memory-related synaptic plasticity, in the rodent hippocampus showed that low n oligomers were the soluble Abeta species primarily responsible for the disruption of synaptic plasticity in vivo. Exogenously applied and endogenously generated anti-Abeta antibodies rapidly neutralized and prevented the synaptic plasticity disrupting effects of these very potent Abeta oligomers. This suggests that active or passive immunotherapeutic strategies for early AD should target Abeta oligomers in the brain. The ability of agents that reduce nitrosative/oxidative stress or antagonize stress-activated kinases to prevent Abeta inhibition of LTP in vitro points to a key role of these cellular mechanisms at very early stages in Abeta-induced neuronal dysfunction. A combination of antibody-mediated inactivation of Abeta oligomers and pharmacological prevention of cellular stress mechanisms underlying their synaptic plasticity disrupting effects provides an attractive strategy in the prevention of early AD. PMID- 16042546 TI - Involvement of presenilins in cell-survival signalling pathways. AB - Familial AD (Alzheimer's disease) is a rare autosomal dominant form of AD, associated with clinical and pathological features similar to those identified in the more prevalent sporadic AD cases. The majority of familial AD cases are caused by mutations in either of the highly homologous PS (presenilins), an essential component of the gamma-secretase enzyme complex, or amyloid precursor protein, a gamma-secretase substrate and the precursor of amyloid beta peptides. The observation that PS are absolutely required for gamma-secretase activity, and parallel studies demonstrating that PS interact with several signalling molecules, modulate their stability or regulate their proteolysis, have led to the suggestion that involvement of PS in additional signalling pathways mediating key cellular functions may contribute to the pathogenesis and progression of neurodegeneration. In this paper, we review PS-regulated molecules, their role in cell signalling and possible involvement in neurodegeneration in patients suffering from AD. PMID- 16042547 TI - Evidence that vitamin D3 reverses age-related inflammatory changes in the rat hippocampus. AB - One of the major challenges in neuroscience is to identify the changes which accompany aging and which contribute to the well-documented age-related deterioration in cognitive function. This is a particular challenge in the light of the vast array of reported changes, which include morphological changes like synaptic and perhaps cell loss, alteration in membrane composition and the resultant changes in function of membrane proteins, modulation of the hypothalamo pituitary axis, impaired calcium homoeostatic mechanisms, alteration in enzyme function and decreased neurotransmitter release. In the past few years, evidence suggesting that an aged brain exhibits signs of oxidative stress and inflammatory stress has been accumulating, and recent evidence using microarray analysis has added support to this view. In this paper, we provide evidence to suggest that vitamin D3 acts as an anti-inflammatory agent and reverses the age-related increase in microglial activation and the accompanying increase in IL-1beta (interleukin-1beta) concentration. PMID- 16042548 TI - Expression of normal sequence pathogenic proteins for neurodegenerative disease contributes to disease risk: 'permissive templating' as a general mechanism underlying neurodegeneration. AB - Loci underlying autosomal dominant forms of most neurodegenerative disease have been identified: prion mutations cause Gerstmann Straussler syndrome and hereditary Creutzfeldt-Jakob disease, tau mutations cause autosomal dominant frontal temporal dementia and alpha-synuclein mutations cause autosomal dominant Parkinson's disease. In these cases, the pathogenic mutation is in the protein that is deposited in the diseased tissue and the whole protein is deposited. In Alzheimer's disease, mutations in amyloid precursor protein or in the presenilins cause autosomal dominant disease. These are the substrate and proteases responsible for the production of the deposited peptide Abeta. Thus, in all the cases, the mutations lead to the disease by a mechanism that involves the deposition process. Furthermore, sporadic forms of all these diseases are predisposed by genetic variability at the same loci, implying that the quantity of the normal protein influences the risk of this form of disease. These results show that the amount of pathogenic protein expression is a key factor in determining disease initiation. Recent work on transgenic models of these diseases is consistent with the view that there are two stages of pathogenesis: a concentration-dependent formation of a pathogenic protein oligomer followed by aggregation on to this oligomeric template by a process that is less dependent on the concentration of the protein. PMID- 16042549 TI - Evidence suggesting that Homo neanderthalensis contributed the H2 MAPT haplotype to Homo sapiens. AB - The tau (MAPT) locus exists as two distinct clades, H1 and H2. The H1 clade has a normal linkage disequilibrium structure and is the only haplotype found in all populations except those derived from Caucasians. The H2 haplotype is the minor haplotype in Caucasian populations and is not found in other populations. It shows no recombination over a region of 2 Mb with the more common H1 haplotype. The distribution of the haplotype and analysis of the slippage of dinucleotide repeat markers within the haplotype suggest that it entered Homo sapiens populations between approx. 10000 and 30000 years ago. However, sequence comparison of the H2 haplotype with the H1 haplotype and with the chimp sequence suggests that the common founder of the H1 and H2 haplotypes was far earlier than this. We suggest that the H2 haplotype is derived from Homo neanderthalensis and entered H. sapiens populations during the co-existence of these species in Europe from approx. 45000 to 18000 years ago and that the H2 haplotype has been under selection pressure since that time, possibly because of the role of this H1 haplotype in neurodegenerative disease. PMID- 16042550 TI - Pathophysiology, pleiotrophy and paradigm shifts: genetic lessons from Parkinson's disease. AB - PD (Parkinson's disease) is an aetiologically heterogeneous disorder characterized by a clinical phenotype consisting of resting tremor, rigidity and bradykinesia. Motor symptoms are associated with a progressive loss of dopaminergic neurons, with Lewy body inclusions within surviving neurons. Although heritability studies have shown evidence of familial aggregation, twin studies have provided limited support for a genetic aetiology. Nevertheless, classical linkage methods have nominated 11 regions of the genome and pathogenic mutations have been identified in several genes, including alpha-synuclein, parkin, ubiquitin C-terminal hydrolase L1, oncogene DJ-1, PTEN-induced protein kinase 1 and microtubule-associated protein tau. Most recently, heterozygous mutations in LRRK2 (leucine-rich repeat kinase 2) were found to cause late-onset, autosomal-dominant PD. Despite their consistent clinical phenotype, family members with LRRK2 mutations can have variable alpha-synuclein and tau pathologies. Lrrk2 is a member of the Roc (Ras of complex proteins) family, with Ras GTPase and MAPKKK (mitogen-activated protein kinase kinase kinase) catalytic domains. Thus its discovery highlights vesicle dynamics and secondary-messenger signalling in disease pathophysiology. To diagnose a disease accurately and effectively treat it, requires an understanding of its molecular pathogenesis. Herein, we provide an overview of the genetics of PD, how these discoveries are revolutionizing long-held beliefs and more importantly how this knowledge may be translated into patient therapy. PMID- 16042551 TI - Mechanisms of memory loss in Abeta and tau mouse models. AB - Although memory loss is the central symptom of Alzheimer's disease, the pathophysiological mechanisms leading to dementia are poorly understood. It is difficult to answer this issue with studies in humans and impossible in cultured cells. Therefore animal models are needed to elucidate the molecular mechanisms leading to dementia. The chief neuropathological changes during Alzheimer's disease, namely neurofibrillary tangles and amyloid plaques, have helped us to determine which molecules to focus upon in the animal models, specifically Abeta (amyloid beta) and tau. This paper presents my perspective on what we have learnt about mechanisms of memory loss from Abeta and tau mouse models of Alzheimer's disease. PMID- 16042552 TI - Clinicopathological features of the tauopathies. AB - Developments in molecular neuropathology have led to protein-based classification systems for neurodegenerative disorders. Key proteins include alpha-synuclein, amyloid and tau. Alternative mRNA splicing and post-translational change, induced by a bewildering variety of protein modifying processes such as phosphorylation and ubiquitination, have generated insights into new mechanisms of selective neuronal degeneration. The task now is to bring these developments in protein chemistry to the clinic, to try to determine whether this biochemical diversity can help in explaining the phenotypic variability that is so typical of neurodegeneration in general. In this review, we will explore the clinicopathological diversity of the tau-related disorders with specific reference to three of the most common tauopathies, frontotemporal dementia (familial and sporadic), progressive supranuclear palsy and corticobasal degeneration. PMID- 16042553 TI - The Arf-like GTPase Arl1 and its role in membrane traffic. AB - Small GTP-binding proteins of the Rab and Arf (ADP-ribosylation factor) families play a central role in the membrane trafficking pathways of eukaryotic cells. The prototypical members of the Arf family are Arf1-Arf6 and Sar1, which have well characterized roles in membrane traffic or cytoskeletal reorganization. However, eukaryotic genomes encode additional proteins, which share the characteristic structural features of the Arf family, but the role of these 'Arf-like' (Arl) proteins is less well understood. This review discusses Arl1, a GTPase that is widely conserved in evolution, and which is localized to the Golgi in all species so far examined. The best-characterized effectors of Arl1 are coiled-coil proteins which share a C-terminal GRIP domain, but other apparent effectors include the GARP (Golgi-associated retrograde protein)/VFT (Vps fifty-three) vesicle-tethering complex and Arfaptin 2. As least some of these proteins are believed to have a role in membrane traffic. Genetic analysis in a number of species has shown that Arl1 is not essential for exocytosis, but rather suggest that it is required for traffic from endosomes to the Golgi. PMID- 16042554 TI - A role of SAND-family proteins in endocytosis. AB - Caenorhabditis elegans has recently been used as an attractive model system to gain insight into mechanisms of endocytosis in multicellular organisms. A combination of forward and reverse genetics has identified a number of new membrane trafficking factors. Most of them have mammalian homologues which function in the same transport events. We describe a novel C. elegans gene sand 1, whose loss of function causes profound endocytic defects in many tissues. SAND 1 belongs to a conserved family of proteins present in all eukaryotic species, whose genome is sequenced. However, SAND family has not been previously characterized in metazoa. Our comparison of C. elegans SAND-1 and its yeast homologue, Mon1p, showed a conserved role of the SAND-family proteins in late steps of endocytic transport. PMID- 16042555 TI - Integrin regulation of membrane domain trafficking and Rac targeting. AB - Integrins are crucial regulators of essential cellular processes such as gene expression, cell proliferation and migration. Alteration of these processes is central to tumourigenesis. Integrin signals mediate anchorage dependence of cell growth, while growth of cancer cells is anchorage-independent. Integrins critically regulate Rho family GTPases, that are also involved in cell-cycle progression and oncogenesis. In addition to their effect on GTP loading, integrins independently control the translocation of GTP-bound Rac to the plasma membrane. This step is essential for Rac binding to effectors. Integrins increase membrane affinity for Rac, leading to RhoGDI dissociation and effector coupling locally, in the vicinity of activated/bound integrins. Integrin-regulated Rac binding sites are within CEMMs (cholesterol-enriched membrane microdomains). Integrins control Rac signalling by preventing the internalization of its binding sites in CEMMs. Integrin regulation of signalling pathways initiated in CEMMs may be important for the spatial control of cell migration and anchorage dependence of cell growth. PMID- 16042556 TI - Insights into biological functions across species: examining the role of Rab proteins in YIP1 family function. AB - The YIP1 family comprises an evolutionarily conserved group of membrane proteins, which share the ability to bind di-prenylated Rab proteins. The biochemical capability of YIP1 family proteins suggests a possible role in the cycle of physical localization of Rab proteins between their cognate membranes and the cytosol. YIP1 is essential for viability in yeast and a deletion of YIP1 can be rescued with the human homologue YIP1A. We have made use of this evolutionary conservation of function to generate a series of mutant alleles of YIP1 to investigate the biological role of Yip1p. Our findings indicate evidence for the participation of Yip1p in both Rab and COPII protein function; at present, we are not able to distinguish between the models that these roles represent, i.e. independent or dependent activities of Yip1p. PMID- 16042557 TI - Membrane curvature and the control of GTP hydrolysis in Arf1 during COPI vesicle formation. AB - The GTP switch of the small G-protein Arf1 (ADP-ribosylation factor 1) on lipid membranes promotes the polymerization of the COPI (coat protein complex I) coat, which acts as a membrane deforming shell to form transport vesicles. Real-time measurements for coat assembly on liposomes gives insights into how the GTPase cycle of Arf1 is coupled in time with the polymerization of the COPI coat and the resulting membrane deformation. One key parameter seems to be the membrane curvature. Arf-GAP1 (where GAP stands for GTPase-activating protein), which promotes GTP hydrolysis in the Arf1-COPI complex is highly sensitive to lipid packing. Its activity on Arf1-GTP increases by two orders of magnitude as the diameter of the liposomes approaches that of authentic transport vesicles (60 nm). This suggests that during membrane budding, Arf1-GTP molecules are progressively eliminated from the coated area where the membrane curvature is positive, but are protected from Arf-GAP1 at the bud neck due to the negative curvature of this region. As a result, the coat should be stable as long as the bud remains attached and should disassemble as soon as membrane fission occurs. PMID- 16042558 TI - RhoGDI-3, a promising system to investigate the regulatory function of rhoGDIs: uncoupling of inhibitory and shuttling functions of rhoGDIs. AB - rhoGDIs (Rho GDP dissociation inhibitors) are postulated to regulate the activity and the localization of small G-proteins of the Rho family by a shuttling process involving extraction of Rho from donor membranes, formation of inhibitory cytosolic rhoGDI/Rho complexes, and delivery of Rho to target membranes. However, the role of rhoGDIs in site-specific membrane targeting or extraction of Rho is still poorly understood. We investigated here the in vivo functions of two mammalian rhoGDIs: the specific rhoGDI-3 and the well-studied rhoGDI-1 (rhoGDI) after structure-based mutagenesis. We identified two sites in rhoGDIs, forming conserved interactions with their Rho target, whose mutation results in the uncoupling of inhibitory and shuttling functions of rhoGDIs in vivo. Remarkably, these rhoGDI mutants were detected at Rho-induced membrane ruffles or protrusions, where they co-localized with RhoG or Cdc42, probably identifying for the first time the site of extraction of a Rho protein by a rhoGDI in vivo. We propose that these mutations act by modifying the steady-state kinetics of the shuttling process regulated by rhoGDIs, such that transient steps at the cell membranes now become detectable. They should provide valuable tools for future investigations of the dynamics of membrane extraction or delivery of Rho proteins and their regulation by cellular partners. PMID- 16042559 TI - A model for Rab GTPase localization. AB - The human genome encodes almost 70 Rab GTPases. These proteins are C-terminally geranylgeranylated and are localized to the surfaces of distinct membrane-bound compartments in eukaryotic cells. This mini review presents a working model for how Rabs achieve and maintain their steady-state localizations. Data from a number of laboratories suggest that Rabs participate in the generation of macromolecular assemblies that generate functional microdomains within a given membrane compartment. Our data suggest that these complex interactions are important for the cellular localization of Rab proteins at steady state. PMID- 16042560 TI - Mechanism and role of localized activation of Rho-family GTPases in growth factor stimulated fibroblasts and neuronal cells. AB - Rho-family GTPases regulate various aspects of cell function by controlling cytoskeletal changes; however, their spatial regulation within the cells remains largely unknown. To understand this regulation, we have studied the spatiotemporal activity of Rho-family GTPases in migrating cells and growth factor-stimulated cells by using probes based on the principle of fluorescence resonance energy transfer. In migrating fibroblasts and epithelial cells, the level of RhoA activity is high both at the contractile tail and at the leading edge, whereas Rac1 and Cdc42 activities are high only at the leading edge. In cells stimulated with epidermal growth factor or nerve growth factor, activities of Rac1 and Cdc42 were transiently elevated in a broad area of the plasma membrane, followed by a localized activation at nascent lamellipodia. In contrast, on epidermal growth factor stimulation, RhoA activity decreased diffusely at the plasma membrane. Notably, RhoA activity persisted at the tip of growth factor-induced membrane ruffles and, in agreement with this finding, RhoA is required for membrane ruffling. These observations suggest that the activities of Rho-family GTPases are elaborately regulated in a time- and space-dependent manner to control cytoskeletal changes and that the basic mechanism of controlling cell shape via Rho-family GTPases is common to various cell types. PMID- 16042561 TI - On the action of Brefeldin A on Sec7-stimulated membrane-recruitment and GDP/GTP exchange of Arf proteins. AB - Arf (ADP-ribosylation factor) proteins form a special class of small GTP-binding proteins in that their activation by GDP/GTP exchange is coupled to their recruitment to membranes using a built-in structural mechanism. These coupled processes are stimulated by GEFs (guanine nucleotide-exchange factors) that carry a catalytic Sec7 domain, whose basic mechanism has been uncovered by biochemical and structural studies. Crystal structures of intermediates of the GDP/GTP exchange reaction, from which GDP has not dissociated, notably allowed a movie of the exchange reaction to be reconstituted. They showed that Sec7 domains secure Arf-GDP to membranes before they proceed to nucleotide dissociation, and thus are active participants to the coupling of membrane-recruitment to nucleotide exchange. The drug BFA (Brefeldin A) was used to trap the complex that initiates the exchange reaction, providing a structural basis for its inhibition of Arf and its action on the membrane-recruitment of isolated Sec7 domains. Based on the dissection of this basic mechanism, the survey of reported BFA effects in cells on large multidomain ArfGEFs of the BIG1/2 and GBF1 families shows that the levels and compartmental distribution of BFA-induced recruitment of ArfGEFs to membranes cannot be explained from isolated Sec7 domains acting as independent domains. This leads to the hypothesis that Sec7 activity is inhibited in these ArfGEFs by an intramolecular interaction, which would be released by interaction with a compartment-specific receptor. PMID- 16042562 TI - Localization and function of Arf family GTPases. AB - Arfs are a family of Ras-related GTP-binding proteins that function in the regulation of membrane trafficking and structure. The six mammalian Arf proteins are expressed ubiquitously and so it is anticipated that each will have a distinct localization and function within the cell. It has been assumed that much of this specificity will be defined by determining which regulators of Arfs, the GEFs (guanine nucleotide-exchange factors) and GAPs (GTPase-activating proteins) function with which Arf proteins. Although in vitro assays may indicate Arf preferences for the numerous Arf GEFs and GAPs that have been identified, in the cell the different Arfs, GEFs and GAPs are targeted to specific compartments where they carry out their functions. We have embarked on studies to define regions of the Arf1 and Arf6 proteins that determine their sites of action and specific activities at the Golgi and plasma membrane respectively. Chimaeras were made between Arf1 and Arf6 in order to identify regions of the protein that contributed to targeting and function. Whereas Arf6 is targeted to the plasma membrane through multiple regions along the protein, we have found a Golgi targeting region in Arf1 that is sufficient to target Arf6 to the Golgi complex. PMID- 16042563 TI - ARL1 has an essential role in Trypanosoma brucei. AB - Myristoyl-CoA protein:NMT (N-myristoyl transferase) catalyses the N myristoylation of cellular proteins with a range of functions and is essential for viability in the protozoan parasites, Leishmania major and Trypanosoma brucei. In our investigations to define the essential downstream targets of NMT, we have focused on the ARF (ADP-ribosylation factor) family of proteins, as growth arrest in Saccharomyces cerevisiae mutants with reduced NMT activity correlates with decreased modification of members of this group of proteins. We have identified nine ARF/ARLs (where ARL stands for ARF-like) encoded in the T. brucei and T. cruzi genomes and ten in L. major. The T. brucei ARL1 protein is expressed only in the mammalian bloodstream form of the parasite, in which it is localized to the Golgi apparatus. RNAi (RNA interference) has been used to demonstrate that ARL1 is essential for viability in these infective cells. Before cell death, depletion of ARL1 protein results in disintegration of the Golgi structure and a delay in exocytosis of the abundant GPI (glycosylphosphatidylinositol)-anchored VSG (variant surface glycoprotein) to the parasite surface. PMID- 16042564 TI - Spatiotemporal regulation of the Pak1 kinase. AB - Pak1 (p21-activated kinase 1) is a key regulator of the actin cytoskeleton, adhesion and cell motility. Such biological roles require a tight spatial and kinetic control of its localization and activity. We summarize here the current knowledge on Pak1 dynamics in vivo. Inactive dimeric Pak1 is mainly cytosolic. Localized interaction with the activators Cdc42-GTP and Rac1-GTP stimulates the kinase at the sites of cellular protrusions. Moreover, Pak1 is dynamically engaged into multiprotein complexes forming adhesions to the extracellular matrix. Cutting edge microscopy technologies on living cells are finally shedding light on the intricate spatiotemporal mechanisms regulating Pak1. PMID- 16042565 TI - Function and regulation of RhoE. AB - The three Rnd proteins, Rnd1, Rnd2 and RhoE/Rnd3, are a subset of Rho family proteins that are unusual in that they bind but do not hydrolyse GTP, and are therefore not regulated by the classical GTP/GDP conformational switch of small GTPases. Increased expression of each Rnd protein induces loss of stress fibres in cultured fibroblasts and epithelial cells, acting antagonistically to RhoA, which stimulates stress fibre formation. RhoE is farnesylated and localizes partly on membranes, including the Golgi and plasma membrane, and in the cytosol. RhoE inhibits RhoA signalling in part by binding to the RhoA-activated serine/threonine kinase ROCK I (Rho-associated kinase I), thereby preventing it from phosphorylating its targets. RhoE activity is itself regulated by phosphorylation by ROCK I on multiple sites. RhoE phosphorylation enhances its stability, leading to an increase in RhoE levels. In addition, phosphorylation reduces its association with membranes and correlates with its ability to induce loss of stress fibres. RhoE also acts independently of ROCK to inhibit cell cycle progression, in part by preventing translation of cyclin D1, and to inhibit transformation of fibroblasts by oncogenic H-Ras. RhoE is therefore a multifunctional protein whose localization and actions are regulated by phosphorylation. PMID- 16042566 TI - Targeting of Rab GTPases to cellular membranes. AB - Rab proteins are members of the superfamily of Ras-like small GTPases and are involved in several cellular processes relating to membrane trafficking and organelle mobility throughout the cell. Like other small GTPases, Rab proteins are initially synthesized as soluble proteins and for membrane attachment they require the addition of lipid moiety(ies) to specific residues of their polypeptide chain. Despite their well-documented roles in regulating cellular trafficking, Rab proteins own trafficking is still poorly understood. We still need to elucidate the molecular mechanisms of their recruitment to cellular membranes and the structural determinants for their specific cellular localization. Recent results indicate that Rab cellular targeting might be Rab dependent, and this paper briefly reviews our current knowledge of this process. PMID- 16042567 TI - Compartmentalized signalling of Ras. AB - Ras proteins associate with cellular membranes by virtue of a series of post translational modifications of their C-terminal CAAX sequences. The discovery that two of the three enzymes that modify CAAX proteins are restricted to the endoplasmic reticulum led to the recognition that all nascent Ras proteins transit endomembranes en route to the PM (plasma membrane) and that at steady state N-Ras and H-Ras are highly expressed on the Golgi apparatus. To test the hypothesis that Ras proteins on internal membranes can signal, we developed a fluorescent probe that reports when and where in living cells Ras becomes active. We found that growth factors stimulated rapid and transient activation of Ras on the PM followed by delayed and sustained activation on the Golgi. We mapped one pathway responsible for this activity as involving PLCgamma (phospholipase Cgamma)/DAG (diacylglycerol)+Ca2+/RasGRP1. Using mammalian cells and fission yeast, we have shown that differential localization of activated Ras preferentially activates distinct signalling pathways. In very recent work, we have found that (i) the subcellular localization of K-Ras can be acutely modulated by phosphorylation of its C-terminal hypervariable region by PKC, (ii) among the membranes upon which phosphorylated K-Ras accumulates is the outer mitochondrial membrane and (iii) phosphorylated, internalized K-Ras promotes apoptosis. Thus the signalling output of Ras depends on its subcellular localization. PMID- 16042568 TI - Use of the Ras binding domain of c-Raf for biochemical and live-cell analysis of Ras activation. AB - Small modular GBDs (GTPase-binding domains) derived from GTPase-effector proteins are useful tools for the selective detection of the active GTP-loaded GTPase conformation, be it in biochemical assays or for imaging purposes. Use of GBD probes requires careful consideration of all features of the GDB-GTPase interaction. It is innate to the strong and specific interaction with the GTP loaded GTPase, that GBDs will protect their partner GTPases from GAP (GTPase activating protein) action. This feature is likely to cause an increase in cellular Ras-GTP levels, in particular in leucocytes and other cells with high steady-state Ras-GDP/GTP cycling rates. By the same token, high levels of GBD expression will interrupt GTPase-initiated signalling, with implications for the activation of the very same GTPase since feedback regulatory mechanisms can impinge on this process. PMID- 16042569 TI - Studying the consequences of immediate loss of gene function in the intestine: APC. AB - The use of mouse models to study neoplasia is proving particularly powerful in dissecting the mechanisms underlying disease initiation and progression. However, the majority of these models have been somewhat limited in studying the very early effects of loss of gene function, as tumour initiation relies upon either constitutive loss of gene function or spontaneous somatic loss of function. We have therefore adopted a strategy of using an inducible Cre-lox-based system to analyse the effects of loss of gene function, the use of which is reviewed here for the intestinal tumour suppressor APC (adenomatous polyposis coli). Using this approach, we have conditionally and synchronously inactivated APC in virtually all the epithelial cells of the adult murine small intestine. After 5 days following induction of Cre-mediated recombination, mice show grossly altered crypt/villus architecture. Deficiency in APC perturbs migration, alters the normal programme of differentiation and results in increased proliferation and apoptosis. Microarray analysis reveals the transcriptome to be significantly altered; reflecting both gross phenotypic changes and changes in transcriptional activation. These findings demonstrate that APC is indeed the critical determinant of cell fate in the intestinal epithelium, explaining its role as the cellular 'gatekeeper' in preventing neoplasia. PMID- 16042570 TI - Prospects in NSAID-derived chemoprevention of colorectal cancer. AB - There is strong evidence for an important role for increased COX (cyclo oxygenase)-2 expression and PG (prostaglandin) E2 production in colorectal tumorigenesis. PGE(2) acts through four E-prostanoid receptors (EP1-4). COX-2 has therefore become a target for the potential chemoprevention and therapy of colorectal cancer. However, any therapeutic/preventive strategy has the potential to have an impact on physiological processes and hence result in side effects. General COX (COX-1 and -2) inhibition by traditional NSAIDs (non-steroidal anti inflammatory drugs), such as aspirin, although chemopreventive, has some side effects, as do some conventional COX-2-selective NSAIDs. As PGE2 is thought to be the major PG species responsible for promoting colorectal tumorigenesis, research is being directed to a number of protein targets downstream of COX-2 that might allow the selective inhibition of the tumour-promoting activities of PGE2, while minimizing the associated adverse events. The PGE synthases and E-prostanoid receptors (EP1-4) have therefore recently attracted considerable interest as potential novel targets for the prevention/therapy of colorectal cancer. Selective (and possibly combinatorial) inhibition of the synthesis and signalling of those PGs most highly associated with colorectal tumorigenesis may have some advantages over COX-2-selective inhibitors. PMID- 16042571 TI - The role of the Wnt signalling pathway in colorectal tumorigenesis. AB - Colorectal cancer (CRC) is the second largest cause of cancer-related deaths in Western countries. CRC arises from the colorectal epithelium as a result of the accumulation of genetic alterations in defined oncogenes and tumour suppressor genes. Mutations in the tumour suppressor APC (adenomatous polyposis coli) genes occur early in the development of CRC and lead to the stabilization of the Wnt pathway component beta-catenin and to the constitutive activation of Wnt signalling. Stabilizing mutations of beta-catenin can also lead to its accumulation, qualifying beta-catenin as a proto-oncogene. Here I will summarize the biochemical interactions occurring in Wnt signalling and describe how alterations in Wnt pathway components lead to CRC. PMID- 16042572 TI - The role of the retinoblastoma protein (Rb) in the nuclear localization of BAG-1: implications for colorectal tumour cell survival. AB - Although the retinoblastoma susceptibility gene RB1 is inactivated in a wide variety of human cancers, the retinoblastoma protein (Rb) has been shown to be overexpressed in colon cancers, which is linked to the anti-apoptotic function of the protein. However, the mechanisms by which Rb regulates apoptosis are yet to be fully elucidated. We have established that Rb interacts with the anti apoptotic BAG-1 (Bcl-2 associated athanogene-1) protein, and that a decrease in nuclear localization of BAG-1 is detectable when the interaction between Rb and BAG-1 is disrupted by expression of the E7 viral oncoprotein. Interestingly, although reported as deregulated in colorectal cancers, we have found that BAG-1 expression is also altered in small adenomas, where its localization was found to be predominantly nuclear. In addition, we have established that maintenance of high nuclear BAG-1 in vitro increases the resistance of adenoma-derived cells to gamma-radiation-induced apoptosis. Our work suggests a novel function for Rb, involving modulation of the subcellular localization of BAG-1. We have found predominant nuclear BAG-1 localization in small adenomas, and suggest that BAG-1 may promote colorectal tumour cell survival by making colonic epithelial cells less sensitive to DNA damage. PMID- 16042573 TI - MutYH (MYH) and colorectal cancer. AB - MAP (MutYH-associated polyposis) is a recently described colorectal adenoma and carcinoma predisposition syndrome that is associated with biallelic-inherited mutations of the human MutY homologue gene, MutYH. MutYH is often also termed MYH. MAP tumours display a mutational signature of somatic guanine-to-thymine transversion mutations in the adenomatous polyposis coli and K-ras genes, reflecting the normal role of MutYH in the base excision repair of adenines misincorporated opposite 7,8-dihydro-8-oxoguanine, a prevalent and stable product of oxidative damage to DNA. However, the full genetic pathway of MAP tumorigenesis has not been elucidated. PMID- 16042574 TI - Epigenetic events in the colorectum and in colon cancer. AB - Colon cancers arise from benign neoplasms and evolve into adenocarcinomas through a stepwise histological progression sequence, proceeding from either adenomas or hyperplastic polyps/serrated adenomas. Genetic alterations have been associated with specific steps in this polyp-adenocarcinoma sequence and are believed to drive the histological progression of colon cancer. Recently, epigenetic alterations, which include CGI (CpG island) DNA methylation, have been shown to occur in colon polyps and colon cancer. The aberrant methylation of genes appears to co-operate with the genetic alterations to drive the initiation and progression of colon polyps to colon cancer. CGI DNA methylation is an epigenetic mechanism that represses gene transcription in normal cellular processes, but it becomes excessive and aberrant in many neoplasms. The aberrant DNA methylation affects CpG-rich regions, called CGIs, in the 5' region of genes and results in transcriptional silencing through effects on transcription factor binding and associated changes in chromatin structure. These hypermethylated genes are not only probable pathogenic events affecting colon-cancer formation, but also neoplasm-specific molecular events that may be useful as molecular markers for colon tumours. Furthermore, aberrant DNA methylation of tumour-suppressor genes may occur secondary to a genetic predisposition or to a field-cancerization effect in the colon and may be useful as molecular markers for the risk of developing colon cancer. PMID- 16042575 TI - Functional consequences of DNA mismatch repair missense mutations in murine models and their impact on cancer predisposition. AB - Mutations in MMR (DNA mismatch repair) genes underlie HNPCC (hereditary non polyposis colon cancer) and also a significant proportion of sporadic colorectal cancers. MMR maintains genome stability and suppresses tumour formation by correcting DNA replication errors and by mediating an apoptotic response to DNA damage. Analysis of mouse lines with MMR missense mutations demonstrates that these MMR functions can be separated and allows the assessment of their individual roles in tumour suppression. These studies in mice indicate that, although the increased mutation rates caused by MMR defects are sufficient to drive tumorigenesis, both functions co-operate in tumour suppression. PMID- 16042576 TI - Relationship between the role of the adenomatous polyposis coli protein in colon cancer and its contribution to cytoskeletal regulation. AB - A unique feature of colon cancer is that truncation mutations in the APC (adenomatous polyposis coli) gene are common to most tumours. The high penetrance of APC mutations, especially in gut epithelium, supports the idea that APC may be involved in a number of the processes that govern the normal maintenance of this tissue: differentiation, migration, proliferation and apoptosis. Indeed, APC is involved in the regulation of beta-catenin and it also is an important regulator of the cytoskeleton. Thus mutations in APC lead to the accumulation of beta catenin, which causes changes in differentiation, and they also produce changes in cytoskeletal organization, which results in altered cell migration and disrupted mitotic spindles. The function of APC in cytoskeletal organization is related to its effect on microtubules and F-actin. Depleting APC from cultured cells leads to changes in cytoskeletal organization. In addition, N-terminal fragments of APC, like those commonly found in tumours, compromise cell migration in Dictyostelium and in early developing chicken embryos. Consistent with the idea that such dominant effects are normally balanced by interactions within the full-length molecule, protein interactions of N-terminal fragments expressed in tumour cells can be altered by binding to C-terminal regions of APC commonly lost in tumours. This review summarizes effects of APC on the cytoskeleton and discusses how these functions of APC may contribute to its role in cancer. PMID- 16042577 TI - Pro-inflammatory mediator leukotriene D4 induces transcriptional activity of potentially oncogenic genes. AB - The inflammatory mediator LTD4 (leukotriene D4) is present at high levels in many inflammatory conditions, and areas of chronic inflammation have an increased risk for subsequent cancer development. We demonstrate here that following LTD4 stimulation, beta-catenin is translocated to the nucleus, triggering the transcriptional activity of the TCF (T-cell factor)/LEF (lymphoid enhancer factor) family of transcription factors. These events are dependent on phosphoinositide-3 kinase activation and glycogen synthase kinase inhibition. Our data suggest that, similar to Wnt signalling, LTD4 increases free beta-catenin and targets it to the nucleus. PMID- 16042578 TI - Molecular targets of nitric-oxide-donating aspirin in cancer. AB - Nitric-oxide-donating aspirin (NO-ASA), consisting of ASA (aspirin) plus an -ONO2 moiety linked to it via a molecular spacer, is a new drug for cancer prevention. NO-ASA seems to overcome the low potency and toxicity of traditional ASA. The ONO2 moiety is responsible for releasing NO, and it appears to be required for biological activity. In studies in vitro, NO-ASA inhibits the growth of colon, pancreatic, prostate, lung, skin, leukaemia and breast cancer cells, and is up to 6000-fold more potent than traditional ASA. This effect is owing to cell kinetics [inhibition of proliferation, induction of apoptosis (multiple criteria) and blocking the G1 to S cell-cycle transition] and cell signalling [inhibition of Wnt signalling (IC50=0.2 microM), inhibition of NF-kappaB (nuclear factor kappaB) activation (IC50=7.5 microM), inhibition of nitric oxide synthase-2 expression (IC50=48 microM), inhibition of MAPK (mitogen-activated protein kinase) signalling (IC50=10 microM) and induction of cyclo-oxygenase-2 at approx. 10 microM]. In studies in vivo, NO-ASA inhibits intestinal carcinogenesis in Min mice (tumour multiplicity was reduced by 59% after 3 weeks, with no effect in control animals and no side effects) and in the N-nitrosobis(2-oxopropyl)amine model of pancreatic cancer, where there was an 89% reduction in NO-ASA (3000 p.p.m. in the diet)-treated animals (P<0.001). There was no statistically significant effect by traditional ASA at equimolar doses. Our data indicate that NO-ASA is a highly promising agent for the prevention and/or treatment of cancer. PMID- 16042579 TI - Design of a MUC1-based cancer vaccine. AB - The epithelial type 1 transmembrane mucin MUC1 is long-established as a marker for monitoring recurrence of breast cancer, and beyond its diagnostic marker qualities, it is a promising target for immunotherapeutic strategies to treat cancer by active specific immunization. The mucin is able to break tolerance and to induce humoral immune responses in healthy subjects and in cancer patients, but the response is generally weak. These natural responses to tumour-associated MUC1 glycoforms indicate that antibody reactivities are more directed to glycopeptide than to non-glycosylated peptide epitopes. To overcome the weak immunogenicity of heavily O-glycosylated MUC1, the question of whether O-linked glycans remain intact during processing in the MHC class II pathway was addressed. Attempts were made to define site-specific O-glycosylation and the structural requirements for efficient endosomal proteolysis by cathepsin L in dendritic cells. A fraction of glycopeptides survive the processing machinery, and have the capacity to bind to MHC class II and to activate sub-populations of glycopeptide-specific helper T-cell clones as a prerequisite for strong and long lasting immune responses to MUC1-positive tumours. Moreover, studies on clusters of sequence-variant repeats, which are interspersed in the repeat domain of MUC1 at high frequency, have revealed that a limited set of concerted amino-acid replacements (Asp-Thr0-Arg1-Pro10 to Glu-Ser0-Arg1-Ala10) contributes considerably to increased peptide flexibility and to under-glycosylation of sequence-variant repeats which in concert modify immunological features of the mucin. Peptides and glycopeptides with the immunodominant DTR (Asp-Thr-Arg) or with the variant ESR (Glu-Ser-Arg) motif, and highly immunogenic peptides of the degenerate repeats that flank the repeat domain are currently evaluated as potential targets in multi-epitopic adjuvant-based vaccine strategies for their capacity to induce cytotoxic T-cell responses. PMID- 16042580 TI - Methylation of the ESR1 CpG island in the colorectal mucosa is an 'all or nothing' process in healthy human colon, and is accelerated by dietary folate supplementation in the mouse. AB - ESR1 is frequently silenced by CGI (CpG island) methylation, both in human colorectal tumours and, in an age-dependent manner, in healthy mucosa. It is not clear, however, whether methylation of individual cytosines occurs randomly within the epithelial genome, or preferentially within individual cells as an 'all-or-nothing' phenomenon. CGI methylation can be quantified in human DNA residues recovered from faecal samples. We used bisulphite genomic sequencing of human DNA from this source and from a colorectal cancer cell line (SW48) to show that the ESR1 CGI is methylated in an allele-specific manner. This provides support for the 'all or none' mechanism for methylation of this gene, and shows how age-dependent methylation of the ESR1 CGI leads rapidly to silencing of the gene within the cells, and hence the colonic crypt within which it occurs. Preliminary studies with a rodent model suggest the rate of age-dependent methylation of ESR1 is modifiable by dietary folate. PMID- 16042581 TI - Cannabinoids and cancer: potential for colorectal cancer therapy. AB - Despite extensive research into the biology of CRC (colorectal cancer), and recent advances in surgical techniques and chemotherapy, CRC continues to be a major cause of death throughout the world. Therefore it is important to develop novel chemopreventive/chemotherapeutic agents for CRC. Cannabinoids are a class of compounds that are currently used in the treatment of chemotherapy-induced nausea and vomiting, and in the stimulation of appetite. However, there is accumulating evidence that they could also be useful for the inhibition of tumour cell growth by modulating key survival signalling pathways. The chemotherapeutic potential for plant-derived and endogenous cannabinoids in CRC therapy is reviewed. PMID- 16042582 TI - DNA damage hypersensitivity in cells lacking BRCA2: a review of in vitro and in vivo data. AB - Since the discovery of the tumour suppressor BRCA2 (encoded by breast-cancer susceptibility gene 2), cells lacking the fully functional protein have consistently been found to show increased sensitivity to a variety of DNA damaging agents, particularly those that cross-link DNA. In this short review, we will bring together these findings and discuss them in the light of our recent in vivo data in the mouse small intestine, which suggests that deletion of cells lacking Brca2 is necessary to avoid the development of potentially tumorigenic clones in this tissue, a system that may be less effective in the mammary glands of humans with germline mutations in BRCA2. PMID- 16042583 TI - Phenotype associated with recessively inherited mutations in DNA mismatch repair (MMR) genes. AB - The MMR (DNA mismatch repair) system helps to maintain the integrity of the genome. This involves eliminating base-base mismatches and insertion/deletion loops, which can lead to microsatellite instability, as seen in tumour cells. Hereditary non-polyposis colon cancer is the result of dominant mutations in MMR genes, such as MLH1, MSH2 and MSH6. More recently there have been case reports of biallelic mutations in the MMR genes MLH1, MSH2 and PMS2. These result in a distinct autosomal recessive cancer predisposition syndrome. The syndrome is characterized by childhood haematological malignancies, brain tumours and the presence of cafe au lait patches. Second primaries occur frequently in this condition, and survival into adulthood is rare. PMID- 16042584 TI - Action of a library of O-glycosylation inhibitors on the growth of human colorectal cancer cells in culture. AB - O-glycosylation is thought to play a significant role in the regulation of cell growth. However, only limited information is available, and few specific and selective inhibitors have been found. We have synthesized a library of O glycosylation inhibitors based on benzyl-O-N-acetyl-D-galactosamine. These inhibitors were tested with an established series of human colorectal cancer cell lines, which model the adenoma-carcinoma sequence. Cancer cells were incubated with the inhibitors, and examined for cell growth patterns, and cellular and subcellular glycosylation using a range of lectins with confocal microscopy. The specificity of O-glycan inhibition was confirmed for the library, relative to other forms of glycosylation. All inhibitors tested resulted in smaller cell yields. However, a differential effect on O-glycosylation was detected using the lectins showing variation of localization at a subcellular level in the various cell lines. Further differential action of the inhibitor library was observed for apoptosis and on the cell cycle with the cell lines tested. This work demonstrates that O-glycosylation is closely involved in the regulation of cell growth in colorectal cancer cells and that the generation of a library of low molecular-mass inhibitors offers a valuable means of examining this regulation at the molecular level. PMID- 16042585 TI - Is COX-2 a 'collateral' target in cancer prevention? AB - NSAIDs (non-steroidal anti-inflammatory drugs) prevent colon and other cancers. The fact that NSAIDs inhibit the eicosanoid pathway prompted mechanistic drug developmental work focusing on COX (cyclo-oxygenase) and its products. The increased prostaglandin E2 levels and the overexpression of COX-2 in colon and many other cancers provided the rationale for clinical trials with COX-2 inhibitors for cancer prevention or treatment. However, one COX-2 inhibitor has been withdrawn from the market because of cardiovascular side effects, and there are concerns about a class effect. Evidence suggests that COX-2 may not be the only, or the ideal, target for cancer prevention; for example, COX-2 is not expressed in human aberrant crypt foci, the earliest recognizable pre-malignant lesion in the colon; COX-2 is expressed in less than half of the adenomas; in vitro data show that NSAIDs do not require the presence of COX-2 to prevent cancer; in familial adenomatous polyposis, the COX-2 inhibitor, celecoxib, had a modest effect, which was weaker than that of a traditional NSAID; and COX-2 specific inhibitors have several COX-2-independent activities, which may account for part of their cancer-preventive properties. The multiple COX-2-independent targets, and the limitations of COX-2 inhibitors, suggest the need to explore targets other than COX-2. PMID- 16042586 TI - DNA mismatch repair status may influence anti-neoplastic effects of butyrate. AB - HNPCC (hereditary non-polyposis colon cancer) is an autosomal-dominant disorder characterized by early-onset CRC (colorectal cancer). HNPCC is most often associated with mutations in the MMR (mismatch repair) genes hMLH1, hMSH2, hMSH6 or hPMS2. The mutator phenotype of a defective MMR system is MSI (microsatellite instability), which also occurs in approx. 15-25% of sporadic CRC cases, where it is associated with the hypermethylation of the promoter region of hMLH1. Dietary factors, including excessive alcohol consumption, ingestion of red meat and low folate intake, may increase the risk of MSI high tumour development. In contrast, aspirin may suppress MSI in MMR-deficient CRC cell lines. Butyrate, a short-chain fatty-acid end product of carbohydrate fermentation in the colon, shares a number of anti-neoplastic properties with aspirin, including inhibiting proliferation and inducing apoptosis of CRC cells. Recent in vitro studies suggest that physiological concentrations of butyrate (0.5-2 mM) may have more potent anti neoplastic effects in CRC cell lines deficient in MMR, but mechanisms for such a differential response remain to be established. PMID- 16042587 TI - DMBT1 expression and glycosylation during the adenoma-carcinoma sequence in colorectal cancer. AB - The gene DMBT1 (deleted in malignant brain tumour-1) has been proposed to play a role in brain and epithelial cancer, but shows unusual features for a classical tumour-suppressor gene. On the one hand, DMBT1 has been linked to mucosal protection, whereas, on the other, it potentially plays a role in epithelial differentiation. Thus its function in a particular tissue is of mechanistic importance for its role in cancer. Because the former function requires secretion to the lumen and the latter function may depend on its presence in the extracellular matrix, we decided to investigate DMBT1 expression, location and its mode of secretion during malignant transformation in colorectal cancer. Using human colorectal PC/AA cell lines and tissue sections from individual patients, we have examined the expression of DMBT1 and its glycosylation in the adenoma carcinoma sequence leading to the adenocarcinoma phenotype. PMID- 16042588 TI - The importance of colonic butyrate transport to the regulation of genes associated with colonic tissue homoeostasis. AB - The transition from normality to malignancy in colorectal cancer is characterized by alterations in the expression of genes associated with the maintenance of tissue homoeostasis. Butyrate, a product of microbial fermentation of dietary fibre in the colon, is known to regulate a number of genes associated with the processes of proliferation, differentiation and apoptosis of colonic epithelial cells, and, hence, homoeostasis of colonic tissue. We have shown previously that the transport of butyrate into colonocytes is of fundamental importance to butyrate's regulatory ability, and therefore sought to assess the expression profile of butyrate-responsive genes in colon cancer tissue, where the expression of the colonic luminal-membrane butyrate transporter, MCT1 (monocarboxylate transporter 1), is significantly down-regulated. In the present paper, we first employed microarray analysis to assess global changes in butyrate-responsive genes using HT29 human colon carcinoma cells treated with butyrate. There was consistency in the butyrate response of selected genes in two other human colonic cell lines (HCT116 and AA/C1) using quantitative real-time PCR. Furthermore, we report that expression levels of selected butyrate-responsive genes involved in the processes of proliferation, differentiation and apoptosis, are deregulated in colon cancer tissue, correlating with decreased expression of MCT1. These findings support our hypothesis that a reduction in MCT1 expression, and hence butyrate transport, can lead to a reduction in the intracellular butyrate levels required to regulate gene expression. Collectively, our results highlight the important contribution of butyrate transport to the maintenance of tissue homoeostasis and disease prevention. PMID- 16042589 TI - Regulation of tetrapyrrole biosynthesis in higher plants. AB - Plant tetrapyrroles are the most abundant biomolecules on the earth and are cofactors of many apoproteins essential for plant function. The four end-products sirohaem, chlorophyll, haem and phytochromobilin are synthesized by a common branched pathway, which is tightly regulated to ensure a continuous supply to the cognate apoproteins. This may induce strong competition between different branches of the pathway for common substrates. In addition, the intermediates, which are phototoxic, must not be allowed to accumulate in the cell. The major control points are during the synthesis of the initial precursor, ALA (5 aminolaevulinic acid), and at the branch points for the insertion of metal ions into the porphyrin macrocycle. A recent study has also suggested that tetrapyrroles are involved in the communication between the chloroplast and the nucleus, strengthening the necessity for tight regulation. However, intermediates remain difficult to quantify mainly due to their low content and the different properties of the intermediates. In this paper, we summarize the regulation of this pathway and we detail why it is important to have an accurate method for the determination of tetrapyrroles in plants. PMID- 16042590 TI - Pantothenate biosynthesis in higher plants. AB - Pantothenate (vitamin B5) is a water-soluble vitamin essential for the synthesis of CoA and ACP (acyl-carrier protein, cofactors in energy yielding reactions including carbohydrate metabolism and fatty acid synthesis. Pantothenate is synthesized de novo by plants and micro-organisms; however, animals obtain the vitamin through their diet. Utilizing our knowledge of the pathway in Escherichia coli, we have discovered and cloned genes encoding the first and last enzymes of the pathway from Arabidopsis, panB1, panB2 and panC. It is unlikely that there is a homologue of the E. coli panD gene, therefore plants must make beta-alanine by an alternative route. Possible candidates for the remaining gene, panE, are being investigated. GFP (green fluorescent protein) fusions of the three identified plant enzymes have been generated and the subcellular localization of the enzymes studied. Work is now being performed to elucidate expression patterns of the transcripts and characterize the proteins encoded by these genes. PMID- 16042591 TI - Flavocytochrome P450 BM3: an update on structure and mechanism of a biotechnologically important enzyme. AB - Since its discovery in the 1980s, the fatty acid hydroxylase flavocytochrome P450 (cytochrome P450) BM3 (CYP102A1) from Bacillus megaterium has been adopted as a paradigm for the understanding of structure and mechanism in the P450 superfamily of enzymes. P450 BM3 was the first P450 discovered as a fusion to its redox partner--a eukaryotic-like diflavin reductase. This fact fuelled the interest in soluble P450 BM3 as a model for the mammalian hepatic P450 enzymes, which operate a similar electron transport chain using separate, membrane-embedded P450 and reductase enzymes. Structures of each of the component domains of P450 BM3 have now been resolved and detailed protein engineering and molecular enzymology studies have established roles for several amino acids in, e.g. substrate binding, coenzyme selectivity and catalysis. The potential of P450 BM3 for biotechnological applications has also been recognized, with variants capable of industrially important transformations generated using rational mutagenesis and forced evolution techniques. This paper focuses on recent developments in our understanding of structure and mechanism of this important enzyme and highlights important problems still to be resolved. PMID- 16042592 TI - Flavoenzyme catalysed oxidation of amines: roles for flavin and protein-based radicals. AB - Amines are a carbon source for the growth of a number of bacterial species and they also play key roles in neurotransmission, cell growth and differentiation, and neoplastic cell proliferation. Enzymes have evolved to catalyse these reactions and these oxidoreductases can be grouped into the flavoprotein and quinoprotein families. The mechanism of amine oxidation catalysed by the quinoprotein amine oxidases is understood reasonably well and occurs through the formation of enzyme-substrate covalent adducts with TPQ (topaquinone), TTQ (tryptophan tryptophylquinone), CTQ (cysteine tryptophylquinone) and LTQ (lysine tyrosyl quinone) redox centres. Oxidation of amines by flavoenzymes is less well understood. The role of protein-based radicals and flavin semiquinone radicals in the oxidation of amines is discussed. PMID- 16042593 TI - Tetrahydrofolate biosynthesis and distribution in higher plants. AB - One-carbon transfer reactions are mediated by H4F (tetrahydrofolate), a soluble coenzyme (vitamin B9) that is synthesized de novo by plants and microorganisms, and absorbed from the diet by animals. H4F synthesis in plants is quartered between the plastids, the cytosol and the mitochondria, a spatial distribution that is not observed in the other organisms and that suggests a complex intracellular traffic. Also, the activity of H4F synthesis fluctuates during plant growth, depending on the tissue and the developmental stage of the seedling, thus illustrating the flexibility of one-carbon metabolism in these organisms. This paper will focus on our recent knowledge about H4F synthesis in the plant cell and will briefly describe the activity of the pathway during the growth and development of the seedling. PMID- 16042594 TI - Mechanistic and inhibition studies of chorismate-utilizing enzymes. AB - The shikimate biosynthetic pathway is utilized in algae, higher plants, bacteria, fungi and apicomplexan parasites; it involves seven enzymatic steps in which phosphoenolpyruvate and erythrose 4-phosphate are converted into chorismate. In Escherichia coli, five chorismate-utilizing enzymes catalyse the synthesis of aromatic compounds such as L-phenylalanine, L-tyrosine, L-tryptophan, folate, ubiquinone and siderophores such as yersiniabactin and enterobactin. As mammals do not possess such a biosynthetic system, the enzymes involved in the pathway have aroused considerable interest as potential targets for the development of antimicrobial drugs and herbicides. As an initiative to investigate the mechanism of some of these enzymes, we showed that the antimicrobial effect of (6S)-6 fluoroshikimate is the result of irreversible inhibition of 4-amino-4 deoxychorismate synthase by 2-fluorochorismate. Based on this study, a catalytic mechanism for this enzyme was proposed, in which the residue Lys-274 is involved in the formation of a covalent intermediate. In another study, Yersinia enterocolitica Irp9, which is involved in the biosynthesis of the siderophore yersiniabactin, was for the first time biochemically characterized and shown to catalyse the formation of salicylate from chorismate via isochorismate as a reaction intermediate. A three-dimensional model for this enzyme was constructed that will guide the search for potent inhibitors of salicylate formation, and hence of bacterial iron uptake. PMID- 16042595 TI - Biophysical tools to monitor enzyme-ligand interactions of enzymes involved in vitamin biosynthesis. AB - Knowledge of biomolecular interactions is of importance to our understanding of biological processes such as enzyme catalysis and inhibition. Biophysical techniques enable sensitive detection and accurate characterization of binding and are therefore powerful tools in enzymology and rational drug design. The applications of NMR spectroscopy and isothermal titration calorimetry to study enzyme-ligand interactions will be discussed. Recent work on ketopantoate reductase, which catalyses an important step on the biosynthetic pathway to vitamin B5, is used to illustrate the potential of this approach. PMID- 16042596 TI - Studies on thiamine diphosphate-dependent enzymes. AB - The 3-deaza analogue of TPP (thiamine diphosphate), a close mimic of the ylid intermediate, has been synthesized and is an extremely potent inhibitor of a variety of TPP-dependent enzymes, binding much more tightly than TPP itself. Results using deazaTPP complexed with the E1 subunit of PDH (pyruvate dehydrogenase) have led to a novel proposal about the mechanism of this enzyme. The 2-substituted forms of deazaTPP, which mimic other intermediates in the catalytic mechanism, can also be synthesized and 2-(1-hydroxyethyl)deazaTPP is also an extremely potent inhibitor of PDC (pyruvate decarboxylase). Attachment of such 2-substituents is expected to be a way to introduce selectivity in the inhibition of various TPP-dependent enzymes. PMID- 16042597 TI - Crystal structure of DMGO provides a prototype for a new tetrahydrofolate-binding fold. AB - The crystal structure of DMGO (dimethylglycine oxidase) from Arthrobacter globiformis in complex with folate compounds has revealed a novel THF (tetrahydrofolate)-binding fold [Leys, Basran and Scrutton (2003) EMBO J. 22, 4038-4048]. This fold is widespread among folate-binding proteins. The crystal structures of aminomethyltransferase (T-protein), YgfZ and TrmE all reveal similar THF-binding folds despite little similarity in sequence or function. The THF-binding site is highly specific for reduced folate compounds and most members of this fold family enhance the nucleophilic character of the THF N10 position. PMID- 16042598 TI - Structures and reaction mechanisms of riboflavin synthases of eubacterial and archaeal origin. AB - The biosynthesis of one riboflavin molecule requires one molecule of GTP and two molecules of ribulose 5-phosphate as substrates. GTP is hydrolytically opened, converted into 5-amino-6-ribitylamino-2,4(1H,3H)-pyrimidinedione by a sequence of deamination, side chain reduction and dephosphorylation. Condensation with 3,4 dihydroxy-2-butanone 4-phosphate obtained from ribulose 5-phosphate leads to 6,7 dimethyl-8-ribityllumazine. The dismutation of 6,7-dimethyl-8-ribityllumazine catalysed by riboflavin synthase produces riboflavin and 5-amino-6-ribitylamino 2,4(1H,3H)-pyrimidinedione. A pentacyclic adduct of two 6,7-dimethyl-8 ribityllumazines has been identified earlier as a catalytically competent reaction intermediate of the Escherichia coli enzyme. Acid quenching of reaction mixtures of riboflavin synthase of Methanococcus jannaschii, devoid of similarity to riboflavin synthases of eubacteria and eukaryotes, afforded a compound whose optical absorption and NMR spectra resemble that of the pentacyclic E. coli riboflavin synthase intermediate, whereas the CD spectra of the two compounds have similar envelopes but opposite signs. Each of the compounds could serve as a catalytically competent intermediate for the enzyme by which it was produced, but not vice versa. All available data indicate that the respective pentacyclic intermediates of the M. jannaschii and E. coli enzymes are diastereomers. Whereas the riboflavin synthase of M. jannaschii is devoid of similarity with those of eubacteria and eukaryotes, it has significant sequence similarity with 6,7 dimethyl-8-ribityllumazine synthases catalysing the penultimate step of riboflavin biosynthesis. 6,7-Dimethyl-8-ribityllumazine synthase and the archaeal riboflavin synthase appear to have diverged early in the evolution of Archaea from a common ancestor. PMID- 16042599 TI - Isoprenoid biosynthetic pathways as anti-infective drug targets. AB - IPP (isopentenyl diphosphate) and DMAPP (dimethylallyl diphosphate) serve as the universal precursors for the biosynthesis of isoprenoids. Besides the well-known mevalonate pathway, the existence of a second biosynthetic pathway conducive to IPP and DMAPP formation through 1-deoxy-D-xylulose 5-phosphate and 2C-methyl-D erythritol 4-phosphate was discovered approx. 10 years ago in plants and certain eubacteria. It is now known that this pathway is widely distributed in the bacterial kingdom including major human pathogens, such as Mycobacterium tuberculosis and Helicobacter pylori. The pathway is also essential in the malaria vector Plasmodium falciparum. During the last few years, the genes, enzymes, intermediates and mechanisms of the biosynthetic route have been elucidated by a combination of comparative genomics, enzymology, advanced NMR technology and crystallography. The results provide the basis for the development of novel anti-infective drugs. PMID- 16042600 TI - Covalent cofactor attachment to proteins: cytochrome c biogenesis. AB - Haem (Fe-protoporphyrin IX) is a cofactor found in a wide variety of proteins. It confers diverse functions, including electron transfer, the binding and sensing of gases, and many types of catalysis. The majority of cofactors are non covalently attached to proteins. There are, however, some proteins in which the cofactor binds covalently and one of the major protein classes characterized by covalent cofactor attachment is the c-type cytochromes. The characteristic haem binding mode of c-type cytochromes requires the formation of two covalent bonds between two cysteine residues in the protein and the two vinyl groups of haem. Haem attachment is a complex post-translational process that, in bacteria such as Escherichia coli, occurs in the periplasmic space and involves the participation of many proteins. Unexpectedly, it has been found that the haem chaperone CcmE (cytochrome c maturation), which is an essential intermediate in the process, also binds haem covalently before transferring the haem to apocytochromes. A single covalent bond is involved and occurs between a haem vinyl group and a histidine residue of CcmE. Several in vitro and in vivo studies have provided insight into the function of this protein and into the overall process of cytochrome c biogenesis. PMID- 16042601 TI - Biodiversity of cytochrome P450 redox systems. AB - P450s (cytochrome P450 mono-oxygenases) are a superfamily of haem-containing mono oxygenase enzymes that participate in a wide range of biochemical pathways in different organisms from all of the domains of life. To facilitate their activity, P450s require sequential delivery of two electrons passed from one or more redox partner enzymes. Although the P450 enzymes themselves show remarkable similarity in overall structure, it is increasingly apparent that there is enormous diversity in the redox partner systems that drive the P450 enzymes. This paper examines some of the recent advances in our understanding of the biodiversity of the P450 redox apparatus, with a particular emphasis on the redox systems in the pathogen Mycobacterium tuberculosis. PMID- 16042602 TI - Inhibition of 7,8-diaminopelargonic acid aminotransferase by amiclenomycin and analogues. AB - Cis and trans stereoisomers of amiclenomycin, a natural L-amino acid antibiotic, have been prepared using unequivocal routes. By using 1H NMR spectroscopy, the configuration of the six-membered ring of natural amiclenomycin was shown to be cis and not trans as originally proposed. Amiclenomycin and some synthetic analogues with the cis configuration irreversibly inactivate DAPA AT (7,8 diaminopelargonic acid aminotransferase), an enzyme involved in biotin biosynthesis, by forming an aromatic PLP (pyridoxal-5'-phosphate)-inhibitor adduct that is tightly bound to the active site. The following kinetic parameters for the inactivation of Escherichia coli DAPA AT by amiclenomycin were derived: K(I)=2 microM and k(inact)=0.4 min(-1). The structure of the aromatic adduct formed upon inactivation was confirmed by UV-visible spectroscopy, X-ray crystal structure determination and MS. Because Mycobacterium tuberculosis DAPA AT is a potential drug target, this enzyme was cloned, overexpressed and purified to homogeneity for biochemical characterization. PMID- 16042603 TI - Vitamin B12: chemistry and biochemistry. AB - Vitamin B12, the 'antipernicious anaemia factor', is required for human and animal metabolism. It was discovered in the late 1940s and its unique corrin ligand was revealed approx. 10 years later by X-ray crystallography. The B12 coenzymes are cofactors in various important enzymatic reactions and are particularly relevant in the metabolism of anaerobic microorganisms. Microorganisms are the only natural sources of the B12-derivatives, whereas most spheres of life (except for the higher plants) depend on these cobalt corrinoids. PMID- 16042604 TI - Anaerobic synthesis of vitamin B12: characterization of the early steps in the pathway. AB - The anaerobic biosynthesis of vitamin B12 is slowly being unravelled. Recent work has shown that the first committed step along the anaerobic route involves the sirohydrochlorin (chelation of cobalt into factor II). The following enzyme in the pathway, CbiL, methylates cobalt-factor II to give cobalt-factor III. Recent progress on the molecular characterization of this enzyme has given a greater insight into its mode of action and specificity. Structural studies are being used to provide insights into how aspects of this highly complex biosynthetic pathway may have evolved. Between cobalt-factor III and cobyrinic acid, only one further intermediate has been identified. A combination of molecular genetics, recombinant DNA technology and bioorganic chemistry has led to some recent advances in assigning functions to the enzymes of the anaerobic pathway. PMID- 16042605 TI - Aerobic synthesis of vitamin B12: ring contraction and cobalt chelation. AB - The aerobic biosynthetic pathway for vitamin B12 (cobalamin) biosynthesis is reviewed. Particular attention is focused on the ring contraction process, whereby an integral carbon atom of the tetrapyrrole-derived macrocycle is removed. Previous work had established that this chemically demanding step is facilitated by the action of a mono-oxygenase called CobG, which generates a hydroxy lactone intermediate. This mono-oxygenase contains both a non-haem iron and an Fe-S centre, but little information is known about its mechanism. Recent work has established that in bacteria such as Rhodobacter capsulatus, CobG is substituted by an isofunctional protein called CobZ. This protein has been shown to contain flavin, haem and Fe-S centres. A mechanism is proposed to explain the function of CobZ. Another interesting aspect of the aerobic cobalamin biosynthetic pathway is cobalt insertion, which displays some similarity to the process of magnesium chelation in chlorophyll synthesis. The genetic requirements of cobalt chelation and the subsequent reduction of the metal ion are discussed. PMID- 16042606 TI - Biotin synthase mechanism: an overview. AB - Biotin synthase, a member of the 'radical SAM' (S-adenosylmethionine) family, converts DTB (dethiobiotin) into biotin. The active form of the Escherichia coli enzyme contains two (Fe-S) centres, a (4Fe-4S) and a (2Fe-2S). The (4Fe-4S)2+/+ mediates the electron transfer required for the reductive cleavage of SAM into methionine and a DOA* (deoxyadenosyl radical). Two DOA*, i.e. two SAM molecules, are consumed to activate the positions 6 and 9 of DTB. A direct transfer of isotope from the labelled substrate into DOAH (deoxyadenosine) has been observed with 2H, although not quantitatively, but not with tritium. The source of the sulphur introduced to form biotin is still under debate. We have shown that the (2Fe-2S)2+ cluster can be reconstituted in the apoenzyme with S2- and Fe2+. When S2- was replaced by [34S2-], [35S2-] or Se2-, biotin containing mostly the sulphur isotopes or selenium was obtained. This leads us to favour the hypothesis that the (2Fe-2S) centre is the sulphur donor, which may explain the absence of turnover of the enzyme. DTBSH (9-mercaptodethiobiotin), which already contains the sulphur atom of biotin, was shown to be an alternative substrate of biotin synthase both in vivo and with a crude extract. When this compound was tested with a well-defined in vitro system, the same turnover of one and similar reaction rates were observed for DTB and DTBSH. We postulate that the same intermediate is formed from both substrates. PMID- 16042607 TI - Molecular and thermodynamic bioenergetics. AB - Studies of biological sciences can be approached in two ways: reductively, as in molecular biology, or holistically, as in systems biology. In this paper, I illustrate my views on approaches to bioenergetics through the analysis of molecular energy transduction and of general thermodynamics relationships in systems biology. The future lies with the second as the first is nearing completion. PMID- 16042608 TI - Is a third proton-conducting pathway operative in bacterial cytochrome c oxidase? AB - Despite the existence of several three-dimensional structures of cytochrome c oxidases, a detailed understanding of pathways involved in proton movements through the complex remains largely elusive. Next to the two well-established pathways (termed D and K), an additional proton-conducting network ('H-channel') has been proposed for the beef heart enzyme. Yet, our recent mutational studies on corresponding residues of the Paracoccus denitrificans cytochrome c oxidase provide no clues that such a pathway operates in the prokaryotic enzyme. PMID- 16042609 TI - Terminal oxidases of cyanobacteria. AB - The respiratory chain of cyanobacteria appears to be branched rather than linear; furthermore, respiratory and photosynthetic electron-transfer chains co-exist in the thylakoid membrane and even share components. This review will focus on the three types of terminal respiratory oxidases identified so far on a genetic level in cyanobacteria: aa3-type cytochrome c oxidase, cytochrome bd-quinol oxidase and the alternative respiratory terminal oxidase. We summarize here their genetic, biochemical and biophysical characterization to date and discuss their interactions with electron donors as well as their physiological roles. PMID- 16042610 TI - Ion translocation by the Escherichia coli NADH:ubiquinone oxidoreductase (complex I). AB - The energy-converting NADH:ubiquinone oxidoreductase, also known as respiratory complex I, couples the transfer of electrons from NADH to ubiquinone with the translocation of ions across the membrane. It was assumed that the complex exclusively works as a proton pump. Recently, it has been proposed that complex I from Klebsiella pneumoniae and Escherichia coli work as Na+ pumps. We have used an E. coli complex I preparation to determine the type of ion(s) translocated by means of enzyme activity, generation of a membrane potential and redox-induced Fourier-transform infrared spectroscopy. We did not find any indications for Na+ translocation by the E. coli complex I. PMID- 16042611 TI - Structure-function relationships in mitochondrial complex I of the strictly aerobic yeast Yarrowia lipolytica. AB - The obligate aerobic yeast Yarrowia lipolytica has been established as a powerful model system for the analysis of mitochondrial complex I. Using a combination of genomic and proteomic approaches, a total of 37 subunits was identified. Several of the accessory subunits are predicted to be STMD (single transmembrane domain) proteins. Site-directed mutagenesis of Y. lipolytica complex I has provided strong evidence that a significant part of the ubiquinone reducing catalytic core resides in the 49 kDa and PSST subunits and can be modelled using X-ray structures of distantly related enzymes, i.e. water-soluble [NiFe] hydrogenases from Desulfovibrio spp. Iron-sulphur cluster N2, which is related to the hydrogenase proximal cluster, is directly involved in quinone reduction. Mutagenesis of His226 and Arg141 of the 49 kDa subunit provided detailed insight into the structure-function relationships around cluster N2. Overall, our findings suggest that proton pumping by complex I employs long-range conformational interactions and ubiquinone intermediates play a critical role in this mechanism. PMID- 16042612 TI - Ubiquinone reduction in the photosynthetic reaction centre of Rhodobacter sphaeroides: interplay between electron transfer, proton binding and flips of the quinone ring. AB - This review is focused on reactions that gate (control) the electron transfer between the primary quinone Q(A) and secondary quinone Q(B) in the photosynthetic reaction centre of Rhodobacter sphaeroides. The results on electron and proton transfer are discussed in relation to structural information and to the steered molecular dynamics simulations of the Q(B) ring flip in its binding pocket. Depending on the initial position of Q(B) in the pocket and on certain conditions, the rate of electron transfer is suggested to be limited either by the quinone ring flip or by the charge-compensating proton equilibration between the surface and the buried Q(B) site. PMID- 16042613 TI - Rewiring photosynthesis: engineering wrong-way electron transfer in the purple bacterial reaction centre. AB - The purple bacterial reaction centre uses light energy to separate charge across the cytoplasmic membrane, reducing ubiquinone and oxidizing a c-type cytochrome. The protein possesses a macroscopic structural two-fold symmetry but displays a strong functional asymmetry, with only one of two available membrane-spanning branches of cofactors (the so-called A-branch) being used to catalyse photochemical charge separation. The factors underlying this functional asymmetry have been the subject of study for many years but are still not fully understood. Site-directed mutagenesis has been partially successful in rerouting electron transfer along the normally inactive B-branch, allowing comparison of the kinetics of equivalent electron transfer reactions on the two branches. Both the primary and secondary electron transfer steps on the B-branch appear to be considerably slower than their A-branch counterparts. The effectiveness of different mutations in rerouting electron transfer along the B-branch of cofactors is discussed. PMID- 16042614 TI - Role of electron-transfer quenching of chlorophyll fluorescence by carotenoids in non-photochemical quenching of green plants. AB - NPQ (non-photochemical quenching) is a fundamental photosynthetic mechanism by which plants protect themselves against excess excitation energy and the resulting photodamage. A discussed molecular mechanism of the so-called feedback de-excitation component (qE) of NPQ involves the formation of a quenching complex. Recently, we have studied the influence of formation of a zeaxanthin chlorophyll complex on the excited states of the pigments using high-level quantum chemical methodology. In the case of complex formation, electron-transfer quenching of chlorophyll-excited states by carotenoids is a relevant quenching mechanism. Furthermore, additionally occurring charge-transfer excited states can be exploited experimentally to prove the existence of the quenching complex during NPQ. PMID- 16042615 TI - Channelrhodopsins: directly light-gated cation channels. AB - Phototaxis and photophobic responses of green algae are mediated by rhodopsins with microbial type chromophores, i.e. all-trans-retinal in the ground state. The green alga Chlamydomonas reinhardtii was recently completely sequenced and the EST (expressed sequence tag) database was made public. We and others detected overlapping partial cDNA sequences that encode two proteins which we termed channelopsins (Chops). The N-terminal half of chop1 (approximately 300 of 712 amino acids) comprises hypothetical seven-transmembrane segments with sequence similarity to the proton pump bacteriorhodopsin and the chloride pump halorhodopsin. Even though the overall sequence homology is low, several amino acids are conserved that define the retinal-binding site and the H+-transporting network in BR (bacteriorhodopsin). Expression of Chop1, or only the hydrophobic core, in Xenopus laevis oocytes, enriched with retinal, produced a light-gated conductance (maximum at approx. 500 nm), which shows characteristics of a channel [ChR1 (channelrhodopsin-1)] that is selectively permeable for protons. Also ChR2 (737 amino acids) is an ion channel that is switched directly by light and also here the hydrophobic N-terminal half of the protein is sufficient to enable light sensitive channel activity. The action spectrum is blue-shifted (maximum at approx. 460 nm) with respect to ChR1. In addition to protons, ChR2 is permeable to univalent and bivalent cations. We suggest that ChRs are involved in phototaxis of green algae. We show that heterologous expression of ChR2 is useful to manipulate intracellular pCa or membrane potential of animal cells, simply by illumination. PMID- 16042616 TI - Active membrane transport and receptor proteins from bacteria. AB - A general strategy for the expression of bacterial membrane transport and receptor genes in Escherichia coli is described. Expression is amplified so that the encoded proteins comprise 5-35% of E. coli inner membrane protein. Depending upon their topology, proteins are produced with RGSH6 or a Strep tag at the C terminus. These enable purification in mg quantities for crystallization and NMR studies. Examples of one nutrient uptake and one multidrug extrusion protein from Helicobacter pylori are described. This strategy is successful for membrane proteins from H. pylori, E. coli, Enterococcus faecalis, Bacillus subtilis, Staphylococcus aureus, Microbacterium liquefaciens, Brucella abortus, Brucella melitensis, Campylobacter jejuni, Neisseria meningitides, Streptomyces coelicolor and Rhodobacter sphaeroides. PMID- 16042617 TI - NMR and fluorescence spectroscopy approaches to secondary and primary active multidrug efflux pumps. AB - Multidrug efflux pumps are found in all major transporter families. Along with a lack of three-dimensional structure information, the mechanism of drug recognition, energy coupling with drug translocation and the catalytic cycle are so far not understood. In the present study, we present first data of a fluorescence-based assay to study the pH-gradient-mediated activity of the multidrug antiporter EmrE, by co-reconstitution with the light-driven proton pump bacteriorhodopsin. In addition to biochemical approaches, the emerging technique, solid-state NMR, can be used for the investigation of these transporters. A number of experiments based on MAS (magic angle sample spinning) NMR are available to provide data on protein structure and dynamics, drug binding and protein-lipid interactions. However, these experiments dictate a number of constraints with respect to sample preparation that will be discussed for proteins from the SMR (small multidrug resistance transporter) family. In addition, 2H-NMR is used to probe protein mobility of Lactococcus lactis ABC transporter, LmrA. PMID- 16042618 TI - Subunit movement in individual H+-ATP synthases during ATP synthesis and hydrolysis revealed by fluorescence resonance energy transfer. AB - F-type H+-ATP synthases synthesize ATP from ADP and phosphate using the energy supplied by a transmembrane electrochemical potential difference of protons. Rotary subunit movements within the enzyme drive catalysis in either an ATP hydrolysis or an ATP synthesis direction respectively. To monitor these subunit movements and associated conformational changes in real time and with subnanometre resolution, a single-molecule FRET (fluorescence resonance energy transfer) approach has been developed using the double-labelled H+-ATP synthase from Escherichia coli. After reconstitution into a liposome, this enzyme was able to catalyse ATP synthesis when the membrane was energized. PMID- 16042619 TI - Structural features and nucleotide-binding capability of the C subunit are integral to the regulation of the eukaryotic V1Vo ATPases. AB - V-ATPases (vacuolar ATPases) are responsible for acidification of intracellular compartments and, in certain cases, proton transport across the plasma membrane of eukaryotic cells. They are composed of a catalytic V1 sector, in which ATP hydrolysis takes place, and the Vo sector, which functions in proton conduction. The best established mechanism for regulating the V-ATPase activity in vivo involves reversible dissociation of the V1 and Vo domains, in which subunit C is intimately involved. In the last year, impressive progress has been made in elucidating the structure of the C subunit and its arrangement inside the V ATPase. Nucleotide occupancy by subunit C, followed by conformational changes of this subunit has shed light on the mechanism of V-ATPase regulation. PMID- 16042620 TI - A study of the horseradish peroxidase catalytic site by FTIR spectroscopy. AB - Vibrational changes in the catalytic site of horseradish peroxidase were investigated by FTIR (Fourier-transform infrared) spectroscopy in the 1000-2500 cm(-1) range. Difference spectra were generated by photolysis of the haemII-CO compound at different pH/pD values. The spectra report on the fine structure around the catalytic site and show vibrational changes of protein backbone, amino acid residues and cofactors. Assignments of the FTIR vibrations can be made based upon known crystal structures, comparisons with absorption frequencies and extinction coefficients of model amino acids and cofactors, effects of H2O/2H2O exchange and changes of pH/pD. Concomitant with the photolysis of the CO ligand are changes due to haem and protein vibrations, predominant among which are arginine and histidine residue vibrations. PMID- 16042621 TI - AHR1B, a new functional aryl hydrocarbon receptor in zebrafish: tandem arrangement of ahr1b and ahr2 genes. AB - The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor that regulates gene expression following activation by TCDD (2,3,7,8 tetrachlorodibenzo-p-dioxin) or a variety of other synthetic and natural compounds. Previous studies have identified two AHR genes, AHR1 and AHR2, in zebrafish (Danio rerio), a widely used model species for studying vertebrate development and an emerging model in developmental toxicology. Zebrafish AHR2 binds TCDD with high affinity, is transcriptionally active and has a major role in mediating the developmental toxicity of TCDD. Zebrafish AHR1 lacks the ability to bind TCDD and activate transcription, and has no known function. In the present study, we report a new zebrafish AHR, designated AHR1B, which shares 34% amino acid sequence identity with AHR1 (AHR1A). The ahr1b gene resides on chromosome 22, adjacent to ahr2, whereas the ahr1a gene is located on chromosome 16. AHR1B is expressed in embryos as early as 24 hours post-fertilization and increases through the next 2 days, but expression is not inducible by TCDD. In contrast with the previously identified AHR1A, in vitro-expressed AHR1B protein exhibits specific, high-affinity binding of [3H]TCDD. Furthermore, AHR1B is able to activate the transcription of a reporter gene under the control of AHR response elements with an efficacy comparable with that of AHR2, but with a higher EC50. We speculate that AHR1B may have a physiological role, such as in embryonic development, whereas AHR2 mediates the response to xenobiotics. PMID- 16042622 TI - An AU-rich stem-loop structure is a critical feature of the perinuclear localization signal of c-myc mRNA. AB - In eukaryotic cells, several mRNAs including those of c-myc and c-fos are localized to the perinuclear cytoplasm and associated with the cytoskeleton. The localization element of c-myc mRNA is present within its 3'UTR (3'-untranslated region) but the precise nature of this signal has remained unidentified. Chemical/enzymatic cleavage with RNases (ribonucleases) and lead have identified single-stranded and double-stranded regions in RNA transcripts of nucleotides 194 280 of the c-myc 3'UTR. Combined with computer predicted structure these results indicate that this region folds so that part of it forms a stem-loop structure. A mutation, that has been previously shown to prevent localization, leads to a different secondary RNA structure in this region as indicated by altered cleavage patterns. Competitive gel-retardation assays, using labelled transcripts corresponding to nucleotides 205-280 of c-myc 3'UTR, and fibroblast extracts revealed that the stem-loop region was sufficient for RNA-protein complex formation. In situ hybridization studies in cells transfected with reporter constructs, in which all or parts of the region corresponding to this stem-loop were linked to beta-globin, indicated that this region was sufficient for localization and that deletion of the nucleotides corresponding to the proposed upper-stem or terminal loop prevented localization. Our hypothesis is that an AU rich stem-loop structure within nt 222-267 in the c-myc 3'UTR forms the perinuclear localization signal. Bioinformatic analysis suggests that this signal shares features with 3'UTRs of other localized mRNAs and that these features may represent a conserved form of signal in mRNA localization mechanisms. PMID- 16042623 TI - Opiate detoxification: what are the goals? PMID- 16042624 TI - Time for a change: putting the Transtheoretical (Stages of Change) Model to rest. PMID- 16042625 TI - When popularity outstrips the evidence: comment on West (2005). PMID- 16042626 TI - Theoretical tools for the industrial era in smoking cessation counselling: a comment on West (2005). PMID- 16042627 TI - Weighing the pros and cons of changing change models: a comment on West (2005). PMID- 16042628 TI - Another nail in the coffin of the transtheoretical model? A comment on West (2005). PMID- 16042629 TI - A premature obituary for the transtheoretical model: a response to West (2005). PMID- 16042631 TI - India: alcohol and public health. PMID- 16042632 TI - "Addiction's" conversation with Malcolm H. Lader. PMID- 16042633 TI - International Center for Alcohol Policies (ICAP)'s latest report on alcohol education: a flawed peer review process. PMID- 16042634 TI - ICAP's latest report on alcohol education -- a flawed process. PMID- 16042635 TI - Fingers burnt: a comment on Foxcroft (2005). PMID- 16042636 TI - Research dissemination: the challenge to ensure quality. PMID- 16042637 TI - What to expect from a 'social aspects' organization, and what to expect from school-based alcohol education. PMID- 16042638 TI - The feasibility of smoking reduction: an update. AB - AIM: To update conclusions of a previous review of smoking reduction on the extent to which (1) smokers spontaneously reduce their smoking, (2) smokers who try to quit and fail return to smoking less, (3) smokers can substantially reduce and maintain reductions via pharmacological and behavioral treatments and (4) smokers compensate when they reduce. METHOD: Qualitative systematic review. DATA SOURCES: Systematic computer searches and other methods. STUDY SELECTION: Published and unpublished studies of smokers not trying to stop smoking. We located 13-26 studies for each of the four aims. DATA EXTRACTION: The first author entered data with confirmation by second author. DATA SYNTHESIS: Due to the heterogeneity of methods and necessity of extensive recalculation, a meta analysis was not feasible. RESULTS: Few daily smokers spontaneously reduce. Among those who try to stop smoking and relapse, some return to reduced smoking but whether they maintain this reduction is unclear. Nicotine replacement (and perhaps behavior therapies) can induce smokers not interested in quitting to make significant reductions in their smoking and maintain these over time. Some compensatory smoking occurs with reduction but significant declines in smoke exposure still occur. CONCLUSIONS: These results indicate that reduction is feasible when aided by treatment. Whether reduction should be promoted will depend on the effect of reduction on health outcomes and future cessation. PMID- 16042640 TI - Comparative study of the effectiveness of slow-release morphine and methadone for opioid maintenance therapy. AB - AIMS: Slow-release morphine may represent a much-needed new pharmacological treatment for opioid dependence. DESIGN: In a 14-week randomized, double-blind, double-dummy, cross-over study oral slow-release morphine was compared with methadone as a treatment for opioid dependency. During two study periods, each consisting of a 1-week titration and a 6-week fixed-dose treatment phase, medication was administered daily under supervised conditions. SETTING: The study was carried out at the Addiction Clinic, Department of Psychiatry, Medical University Vienna. PARTICIPANTS: Sixty-four subjects (56 males, eight females) with opioid dependence participated in the trial. MEASUREMENTS: Efficacy was evaluated on the basis of retention, use of illicit substances based on urinalysis, extent of drug cravings, withdrawal symptoms and general wellbeing. Safety was assessed on the basis of adverse events and clinical and physical examination. Demographic and baseline characteristics were assessed using the European Addiction Severity Index. FINDINGS: Fifty-five patients (86%) completed the study, with a mean methadone dose of 85 mg and a mean slow-release morphine dose of 680 mg. No significant differences in retention or use of illicit substances (opioids, benzodiazepines, cocaine) were observed, irrespective of treatment group or medication. However, patients receiving slow-release morphine had significantly lower depression (P < 0.001) and anxiety scores (P = 0.008) and fewer physical complaints (P < 0.001). CONCLUSIONS: Oral slow-release morphine is as effective as methadone in the treatment of opioid dependency, with comparable safety and tolerability and a greater benefit on patient wellbeing. Greater pharmaceutical diversity represents a modern development in mainstream medicine. Slow-release morphine might represent a future treatment option that will improve long-term outcomes for this target group. PMID- 16042639 TI - A multi-center randomized trial of buprenorphine-naloxone versus clonidine for opioid detoxification: findings from the National Institute on Drug Abuse Clinical Trials Network. AB - AIMS: The clinical effectiveness of buprenorphine-naloxone (bup-nx) and clonidine for opioid detoxification in in-patient and out-patient community treatment programs was investigated in the first studies of the National Institute of Drug Abuse Clinical Trials Network. DESIGN: Diagnostic and Statistical Manual version IV (DSM IV)-diagnosed opioid-dependent individuals seeking short-term treatment were randomly assigned, in a 2 : 1 ratio favoring bup-nx, to a 13-day detoxification using bup-nx or clonidine. METHODS: A total of 113 in-patients (77 bup-nx, 36 clonidine) and 231 out-patients (157 bup-nx, 74 clonidine) participated. Supportive interventions included appropriate ancillary medications and standard counseling procedures guided by a self-help handbook. The criterion for treatment success was defined as the proportion of participants in each condition who were both retained in the study for the entire duration and provided an opioid-free urine sample on the last day of clinic attendance. Secondary outcome measures included use of ancillary medications, number of side effects reported and withdrawal and craving ratings. FINDINGS: A total of 59 of the 77 (77%) in-patients assigned to the bup-nx condition achieved the treatment success criterion compared to eight of the 36 (22%) assigned to clonidine, whereas 46 of the 157 (29%) out-patients assigned to the bup-nx condition achieved the treatment success criterion, compared to four of the 74 (5%) assigned to clonidine. CONCLUSION: The benefits of bup-nx for opioid detoxification are supported and illustrate important ways in which clinical research can be conducted in community treatment programs. PMID- 16042641 TI - Impact of Federal drug law enforcement on the supply of heroin in Australia. AB - AIMS: To conduct an empirical investigation of the efficacy of law enforcement in reducing heroin supply in Australia. Specifically, this paper addresses the question of whether heroin purity levels in the Australian Capital Territory (ACT) could be predicted by heroin seizures at the national level by the Australian Federal Police (AFP) in the preceding year. DESIGN: We considered two forms of evidence. First, a Bayesian Markov Chain Monte Carlo (MCMC) change-point model was used to discover (a) if there was a substantial increase in heroin seizures by the AFP, (b) when the increase began and (c) whether it occurred after increased funding to the Australian Federal Police for the purpose of drug law enforcement. Second, standard time-series methods were used to ascertain whether fluctuations in heroin seizure weights or the frequency of large-scale seizures after the aforementioned changes in seizure levels predicted fluctuations in heroin purity levels in the ACT after autocorrelation had been removed from the purity series. FINDINGS: A Bayesian MCMC change-point model supported the hypothesis that heroin seizures rapidly increased about a year before the estimated decline in heroin purity and after the increased funding of AFP. The autoregression models suggested that 10-20% of the variance in the residuals of the heroin purity series was predicted by appropriately lagged residuals of the seizure-number and log-weight series, after autocorrelation had been removed. CONCLUSION: The overall results are consistent with the hypothesis that large-scale heroin seizures by the AFP reduce street-level heroin supply a year or so later, although the short-term dynamics suggest an 'opponent' response to residual fluctuations in seizures. To our knowledge, this is first time a connection has been identified between large-scale heroin seizures and street level supply. PMID- 16042642 TI - The impact of borderline personality disorder on 12-month outcomes for the treatment of heroin dependence. AB - AIMS: To determine the effects of borderline personality disorder (BPD) on 12 month treatment retention and outcomes for the treatment of heroin dependence. DESIGN: Longitudinal cohort study. Setting Sydney, Australia. PARTICIPANTS: A cohort of 495 heroin users enrolled in the Australian Treatment Outcome Study. FINDINGS: Criteria for BPD were met by 45% of the cohort. At baseline there were no differences in heroin use, but the BPD group had higher levels of polydrug use, crime, needle risk-taking, more injection-related health problems, higher levels of overdose, poorer psychological health and more extensive suicide histories. At 12 months there was no group difference in the cumulative number of treatment days received, but the BPD group had enrolled in more different treatment episodes. Within both groups reductions had occurred in drug use and drug-related problems, with no differences in heroin use, polydrug use or global physical health at 12 months. After taking into account the effects of treatment on outcome, however, BPD was associated independently with a higher level of needle sharing [odds ratio (OR) 3.21], more injection-related health problems (OR 1.90), a higher likelihood of heroin overdose (OR 1.92), poorer global psychological health (OR 2.43), higher levels of current major depression (OR 3.19) and a higher likelihood of attempted suicide (OR 3.89). While BPD participants showed similar reductions in heroin and other drug use to other patients at 12 months, they continued to exhibit higher levels of risk and harm across a range of outcomes. CONCLUSIONS: Screening would appear to be warranted to identify a group who may overtly respond to treatment in terms of drug use per se, but remain at substantially greater risk. PMID- 16042643 TI - Levo-alpha-acetylmethadol (LAAM) versus methadone: treatment retention and opiate use. AB - AIMS: To compare the effects of levo-alpha-acetylmethadol (LAAM) and methadone maintenance (MM) on treatment retention and abstinence from opiate use. DESIGN: A two-group experimental design with patients randomly assigned (2 : 1 LAAM : MM) to receive LAAM (three doses per week) or methadone (daily dosing). SETTING: A community clinic in Los Angeles, California. PARTICIPANTS: A total of 315 patients seeking LAAM or methadone maintenance. INTERVENTION: LAAM or methadone maintenance, plus ancillary services available to all patients. LAAM and methadone dose levels varied according to clinical judgement. Electrocardiograms were administered to LAAM patients monthly. MEASUREMENTS: Treatment status at 26 week follow-up and number of days retained in treatment, weekly clinical urine tests and 26-week research urine test. FINDINGS: LAAM and methadone patients did not differ on treatment retention. LAAM patients were less likely to test positive for opiate use during treatment (40% versus 60%) and at 26-week follow up (39.8% versus 60.2%). Benefits of LAAM were confined to patients (n = 204) still in treatment at 26 weeks (33% positive in patients receiving LAAM and 61% in patients receiving methadone). No adverse events, cardiological or otherwise, were observed with LAAM administration. CONCLUSIONS: LAAM is an effective medication for the treatment of opiate dependence with clinical advantages due not only to the reduction of opiate use but also to the alternate-day dosing schedule. LAAM may be more effective than methadone in promoting abstinence from opiate use among patients for whom LAAM is an acceptable alternative to methadone. PMID- 16042644 TI - Five-year trends in patterns of drug use among people who use stimulants in dance contexts in the United Kingdom. AB - AIMS: To describe and evaluate trends in the use of stimulant drugs over a 5-year period using an under-studied data collection method. DESIGN: Repeated-measures cross-sectional survey. SETTING AND PARTICIPANTS: Annual magazine-based survey targeting people who use stimulant drugs in dance contexts. MEASUREMENTS: Life time use prevalence (ever used), age of first use, current use prevalence (any use within the last month) and extent of use within the last month (number of days used) for a range of stimulant drugs. Additional measures of quantity of ecstasy used were also collected. FINDINGS: Trends in life-time and current prevalence over time have been detected and comparisons made between different stimulant drugs. Evidence is obtained of broad stability in patterns of stimulant use in respect of age of first use and frequency of use among ongoing users. Despite an apparent reduction in the current prevalence of ecstasy use, the proportion of heavy users (usually >4 pills per session) has more than doubled between 1999 and 2003. CONCLUSIONS: This purposively sampled population study has yielded time trend data broadly consistent with other indicators, where they exist, and also has demonstrable potential to identify new drug trends. Further comparisons of purposive samples and randomly formed samples are needed. PMID- 16042645 TI - Alcohol and coronary heart disease risk--is there an unknown confounder? AB - AIMS: To evaluate whether confounding by several known or suspected coronary heart disease risk factors are likely to explain the lower coronary heart disease risk among light alcohol drinkers compared with never-drinkers. DESIGN: A population-based cross-sectional study. METHODS: Hypertension, body mass index (BMI), diabetes, depression, sleep disturbances, smoking, physical activity, life satisfaction, psychological distress, trait anxiety, independent and dependent life events, length of working hours, job control, job strain and effort-reward imbalance were compared between never-drinkers and light drinkers (<70 g of alcohol per week). Data on 41,099 participants (6,222 men, 34,877 women) were derived from two ongoing cohort studies, the '10-Town Study' and 'Finnish Hospital Personnel Study', in Finland in 2000-02. RESULTS: Of the 16 comparisons under study, seven showed significant differences between never-drinkers and light drinkers. Five of the differences favoured never-drinkers and two showed a disadvantage. The latter were low BMI and low leisure-time physical activity, both more common among never-drinkers than among light drinkers. In contrast, smoking, sleep disturbances, trait anxiety, effort-reward imbalance and dependent life events were less common among never-drinkers than among light drinkers. CONCLUSIONS: None of the risk factors studied was a likely candidate for an unknown confounder. PMID- 16042646 TI - Alcohol price elasticities in control and license states in the United States, 1982-99. AB - AIM: The demand for alcohol has been demonstrated repeatedly to be sensitive to price changes. However, estimated price elasticities vary by study region and over time. One explanation for these variations might be that different countries or parts of countries have had different alcohol control systems. The hypothesis addressed in this study was that a regulated market leads to higher transaction costs associated with purchasing alcohol, which in turn increases the full price of the beverages (the nominal cash price plus transaction costs). As a result, the cash price of alcohol represents a smaller part of the full price in a highly regulated market. Assuming that customers respond primarily to changes in full price, the demand for alcohol should be less sensitive to changes in cash price where regulation is stricter. This study examined whether variations in price elasticities were a function of the different regulatory systems in control and license states in the United States during the period 1982-99. DATA AND METHODS: Time-series cross-sectional analyses (in 50 states over 18 years) were conducted. Elasticities were estimated using a multiplicative model based upon first differences of time-series within states. Disposable income and other socio demographic variables were used as control variables. All data were obtained from archival sources. FINDINGS: The demand for spirits and beer were significantly more sensitive to price changes in license states than in control states. The estimated price elasticity for wine sales was also somewhat larger in license states, but not significantly so. CONCLUSION: The lower price elasticities for spirits and beer in the control states support the hypothesis that customers respond primarily to changes in the full price of alcohol. PMID- 16042647 TI - European Union scientific production on alcohol and drug misuse (1976-2000). AB - BACKGROUND: Alcohol and drug misuse is a social and health phenomenon of great relevance in the European Union (EU). One indicator of scientific production in a given area is the analysis of publications included in bibliographic databases. Scientific production on alcohol and drug misuse was analysed in EU member countries, and comparisons were made between countries. METHODS: Analysis of articles on alcohol and drug misuse published during the period 1976-2000 by institutions based in a country of the EU, indexed by PsycINFO. RESULTS: A total of 4825 citations was retrieved. Great Britain published 38.6%, while Sweden, Germany and Spain accounted for a further 30%. The articles dealt with drug and alcohol usage (12.8%), substance abuse (53.5%) and drug and alcohol rehabilitation (34.5%). The articles were published in 13 different languages, more than three-quarters being in English. Spanish was the second language, and was followed by French, German, Dutch and Italian. The articles were published in 521 different journals, and 62 of these published more than 10 articles. The journals publishing most were Addiction, Alcohol and Alcoholism and Drug and Alcohol Dependence. Sixty-eight per cent of the articles were signed by more than one author, and the index of collaboration, between 1996 and 2000, was 3.24. DISCUSSION AND CONCLUSIONS: PsycINFO is useful for making comparisons between countries, because it includes the name and country of the institution. The number of publications in the EU on alcohol and drug misuse increased over the quarter-century analysed. The most used language was English, as it also is for PsycINFO as a whole, and a tendency towards its increased use was observed. Classification of the articles by subject by the Classification Code is too general, and makes it difficult to distinguish between the areas it proposes. Production tends to be concentrated in journals dealing specifically with drug dependence and psychiatry. The index of collaboration is similar to that found in other scientific areas. PMID- 16042648 TI - When smokers are resistant to change: experimental analysis of the effect of patient resistance on practitioner behaviour. AB - AIMS: In the field of motivational interviewing, practitioner confrontational behaviour has been associated with lower levels of patient behaviour change. We set out to explore whether resistance to change among smokers affects practitioner confrontational and other behaviours. DESIGN: Experimental manipulation of levels of patient resistance in a role play. SETTING: The study was conducted at the start of a 2-day health behaviour change workshop. PARTICIPANTS: Thirty-two practitioners who had registered for the workshop. INTERVENTION: The practitioners were assigned randomly to interview a standardized patient (actor) who portrayed a smoker who had been briefed to display either high or low levels of resistance to change. MEASUREMENTS: Interviews were audiotaped and transcribed. Practitioners and standardized patients completed interview ratings at the end of each interview. After listening to each taped interview practitioners were assigned a global score for confrontation, empathy and expert instructional style. Interviews were then submitted to a qualitative analysis. FINDINGS: Higher levels of practitioner confrontational behaviour were observed in the high resistance group. This was evident both from the global scores (median 2 versus 0, P = 0.001) and the qualitative analysis. Global scores for empathy and expert instruction were not significantly different. Qualitative analysis also suggests a pervasive negative impact on other practitioner behaviours. CONCLUSIONS: Higher patient resistance probably leads to an increase in confrontational and other negative behaviours in health professionals attempting to promote behaviour change. This challenges important assumptions about the influence of practitioner behaviour on patient behaviour and subsequent health-related outcomes. PMID- 16042649 TI - Antisocial personality disorder is associated with increased severity of gambling, medical, drug and psychiatric problems among treatment-seeking pathological gamblers. AB - AIMS: To evaluate systematically whether pathological gamblers with antisocial personality disorder (ASPD) experience increased severity of gambling, medical, psychiatric, substance use and psychosocial problems compared to pathological gamblers without ASPD. PARTICIPANTS, DESIGN AND MEASUREMENTS: Pathological gamblers (n = 237) entering an out-patient treatment study for pathological gambling completed the ASPD section of the Structured Clinical Interview for Diagnostic and Statistical Manual version IV (DSM-IV) Personality Disorders, California Psychological Inventory-Socialization Scale, Addiction Severity Index (ASI), Brief Symptom Inventory (BSI) and gambling questionnaires. SETTING: Pathological gambling research clinic. FINDINGS: Thirty-nine (16.5%) pathological gamblers met DSM-IV diagnostic criteria for ASPD. Compared to pathological gamblers without ASPD, pathological gamblers with ASPD were younger, more likely to be male and divorced/separated, and had fewer years of education. They also began gambling earlier in life, reported increased severity of gambling, medical and drug problems, and scored higher on the paranoid ideation, somatization and phobic anxiety subscales of the BSI. Further, logistic regression identified male gender, history of illicit drug use and severity of gambling and medical problems as independent predictors of ASPD. CONCLUSIONS: These results underscore the importance of assessing a wide range of behaviors and personality indices, including ASPD, among treatment-seeking pathological gamblers. PMID- 16042650 TI - The role of supply reduction in an evidence-based illicit drug strategy: comments on Smithson et al. (2005). PMID- 16042651 TI - Classic texts revisited. Re: Similarities in Animal and Human Drug-Taking Behavior by Roland R. Griffiths, George E. Bigelow, Jack E. Henningfield, in: Advances in Substance Abuse, Volume 1, JAI Press Inc., 1980, pp. 190. PMID- 16042654 TI - Review: what do we mean by GERD?--definition and diagnosis. AB - Gastro-oesophageal reflux disease (GERD) is a condition in which the gastro oesophageal reflux provokes symptoms or complications. Since the majority of patients with heartburn do not have mucosal breaks, expressions such as 'endoscopy-negative reflux disease' (ENRD), 'non-erosive reflux disease' (NERD), or even 'reflux-like dyspepsia' and 'functional heartburn' are frequently employed despite the lack of consensus concerning their exact meaning. Moreover, definition of a disease does not mean that precise diagnostic criteria exist. Diagnostic approaches to GERD differ considerably between primary and secondary care. The primary care physician's role is to decide, on the basis of symptoms and clinical examination, if it is likely that the patient has some serious problem which requires urgent investigation and intervention. In practice, a symptom-based diagnosis can often be made reliably because heartburn and regurgitation are very specific for GERD. The secondary care physician has to make a full evaluation of an already highly-selected patient and, as far as possible, to make a comprehensive, accurate diagnosis, using whatever investigative tools are required. However, there is no 'gold standard' for the diagnosis of GERD and 24-hour pH monitoring lacks sensitivity in NERD. Recently, impedance-pH monitoring has been introduced and promising results have been reported. However, this new technology needs further validation and technical improvement before being employed in routine clinical investigation. PMID- 16042655 TI - Review article: gastro-oesophageal reflux disease--the extent of the problem. AB - There have been little review data on the period prevalence of heartburn symptoms in the community. We analysed reflux symptom data from a test-and-treat trial that evaluated community Helicobacter pylori levels and added these to data identified from a literature search. We found 31 papers reporting on 77,671 subjects who provided information on the frequency of heartburn symptoms in the community. Pooled data from these papers suggested that, in Western populations, 25% had symptoms at least once per month, 12% had symptoms at least weekly and 5% had heartburn daily. Reflux symptoms were less common in East Asian populations with only 7% having heartburn at least once per month. Heartburn is a significant problem in Western societies. PMID- 16042656 TI - Review article: gastro-oesophageal reflux disease--the health economic implications. AB - For the vast majority of patients with gastro-oesophageal reflux disease appropriate care involves the management of symptoms with lifestyle advice and drugs. However, there is dissension about the appropriate use of endoscopy, whether drugs should be stepped up or down according to potency, how long drugs should be used for, the role of lifestyle advice, and, related to this, the role of patients' lifestyle choices. This exploration of the economics of gastro oesophageal reflux disease reviews its cost burden to the UK, assesses published economic models for their strengths and weaknesses and examines current recommendations for gastro-oesophageal reflux disease management from a socioeconomic perspective. Drugs prescribed predominantly for dyspepsia cost the UK National Health Service a projected pound sterling 625 million in 2004, 7% of the primary care prescribing budget. When general practitioners consultations, endoscopies, over-the-counter sales and sickness absences are included the UK cost rises to pound sterling 1.5 billion: approximately half of this cost can be ascribed to gastro-oesophageal reflux disease. Emphasis upon regular review and stepping down treatment (while maintaining adequate symptom relief) is both clinically appropriate and resource efficient. Other cost-effectiveness issues largely lack objective answers because investment in treatment for gastro oesophageal reflux disease depends upon how much more, at the margin, society wishes to invest for further but diminishing symptom relief. PMID- 16042657 TI - Review article: Helicobacter pylori and gastro-oesophageal reflux disease. AB - Epidemiological studies demonstrate a negative association between Helicobacter pylori infection and gastro-oesophageal reflux disease and its complications. This might represent a protective effect because of the tendency for H. pylori infection to lower gastric acid secretion with advancing age. However, studies of the effect of H. pylori eradication on gastro-oesophageal reflux disease have failed to show any worsening of gastro-oesophageal reflux disease symptoms. Several interactions between H. pylori and proton-pump inhibitor therapy used to treat gastro-oesophageal reflux disease need to be considered. Helicobacter pylori infection improves the control of gastric acidity by proton-pump inhibitors and this produces a small advantage in clinical control of reflux disease. The infection prevents rebound acid hypersecretion occurring when proton pump inhibitor therapy is discontinued. However, concerns have been expressed that the body gastritis induced by proton-pump inhibitor therapy in H. pylori infected subjects might increase the risk of gastric cancer. At present, it is unclear whether H. pylori should be eradicated in gastro-oesophageal reflux disease patients. PMID- 16042658 TI - Review article: quality-of-life issues in gastro-oesophageal reflux disease. AB - The purpose of this review is to explore issues relating to quality of life in gastro-oesophageal reflux disease, examining the range of generic and disease specific instruments available, their applicability and limitations and to overview the effect of gastro-oesophageal reflux disease on quality of life. Whereas instruments have been developed to assist researchers, there is a paucity of reliable instruments for pragmatic use in the clinical setting. The situation is complicated because there is not necessarily a direct correlation between endoscopic findings and symptom severity and non-erosive reflux disease is now recognized as an important manifestation of gastro-oesophageal reflux disease. However, quality-of-life instruments are useful in evaluating the impact of therapies and interventions, although these are limited, particularly in surgical interventions. Impaired quality of life now forms part of a definition of gastro oesophageal reflux disease, the impact of which goes beyond the symptoms alone. While the symptoms themselves have a negative effect on sufferers' lives, there are secondary effects caused by impaired physical, emotional and social functioning on productivity. Non-gastrointestinal problems caused by gastro oesophageal reflux disease also impair quality of life. There is an ongoing need to develop instruments which truly measure the impact of gastro-oesophageal reflux disease and which are readily interpretable to the individual patient and clinician. PMID- 16042659 TI - Review article: role of pepsin and bile in gastro-oesophageal reflux disease. AB - Gastro-oesophageal reflux disease is defined as the presence of symptoms or lesions that can be attributed to the reflux of gastric contents into the oesophagus. Aspiration and prolonged monitoring studies in humans have shown that reflux of gastric contents is comprised of both acid and non-acid components, in healthy as well as diseased people. Methods to monitor the non-acid component of the refluxate are described in detail. Experimental models suggest that synergism between acid and pepsin and conjugated bile acids have the greatest damaging potential for oesophageal mucosa, although unconjugated bile acids may be caustic at a more neutral pH. Human studies are compatible with a synergistic action between acid and duodenogastric reflux in inducing lesions. During prolonged monitoring studies, typical gastro-oesophageal reflux disease symptoms are more related to acid reflux events than to non-acid reflux events. However, symptoms that persist during acid-suppressive therapy are often related to non-acid reflux events. The therapeutic options for the non-acid component of the refluxate, including acid suppression, prokinetics, baclofen and surgery, are discussed. PMID- 16042660 TI - Review article: the long-term use of proton-pump inhibitors. AB - More than 15 years after the launch of omeprazole in 1988, proton-pump inhibitors remain central to the management of acid-suppression disorders and are unchallenged with regard to their efficacy and popularity among doctors and patients. They are considered safe despite early concerns about the possibility of an association with cancer and gastric atrophy; current concerns about long term proton-pump inhibitor therapy are centred mainly on a possible association with fundic gland polyps and between Helicobacter pylori and gastric atrophic changes. Long-term proton-pump inhibitor usage accounts for the majority of the total proton-pump inhibitor usage. Long-term usage is difficult to define and most patients take proton-pump inhibitors non-continuously. Data indicate that a substantial proportion of long-term users do not have a clear indication for their therapy and there is thus room for reduction or rationalization of treatment. Overall, on-demand therapy is more cost-effective than continuous therapy and should be considered wherever possible. PMID- 16042661 TI - Review article: how valuable are proton-pump inhibitors in establishing a diagnosis of gastro-oesophageal reflux disease? AB - Diagnostic studies for gastro-oesophageal reflux disease (endoscopy, pH testing) have significant limitations. A short trial of acid suppression with a proton pump inhibitor is often used in clinical practice as a diagnostic test for gastro oesophageal reflux disease. If there is a significant improvement in symptoms, the patient is considered to have gastro-oesophageal reflux disease. Several studies have evaluated the administration of proton-pump inhibitors as a diagnostic test for gastro-oesophageal reflux disease. A recent meta-analysis found that response to a proton-pump inhibitor had limited value as a diagnostic test. Likelihood ratios for a positive test ranged from 0.45 to 1.86. The positive predictive value of the test ranged from 0.17 to 0.90 and the negative predictive value from 0.17 to 1, depending on the gold standard against which comparisons were made. These results may be explained by the inadequacy of the gold standard against which the proton-pump inhibitor test is compared or by the inability of acid suppression to differentiate between gastro-oesophageal reflux disease, peptic ulcer disease or dyspepsia. Although a trial of acid suppression may have pragmatic value in clinical practice, the proton-pump inhibitor test is not an accurate test for the diagnosis of gastro-oesophageal reflux disease. PMID- 16042662 TI - Review article: extraoesophageal manifestations of gastro-oesophageal reflux disease. AB - Although recent studies suggest that gastro-oesophageal reflux disease may frequently contribute to ear, nose and throat and respiratory diseases, the cause and-effect relationship is far from proven. The review will address this controversial topic emphasizing recent literature raising concerns about the credibility of this association and our tests to make this diagnosis. The author believes these extraoesophageal symptoms suspected to be secondary to gastro oesophageal reflux disease are an unresolved issue, but selective use of aggressive proton-pump inhibitor therapeutic trials may help to resolve this problem in our individual patients. PMID- 16042663 TI - Metameta-analysis. PMID- 16042664 TI - Assessment of the effect of a single oral dose of telithromycin on sotalol induced qt interval prolongation in healthy women. AB - AIMS: Telithromycin belongs to ketolides, a new class of macrolide antibiotics. Macrolides are known to have the potential to prolong QT interval duration. Previous studies have shown that telithromycin did not induce significant QT interval prolongation in healthy subjects compared with placebo. The main objective of this study was to demonstrate the absence of amplification of QT interval prolongation induced by sotalol, when telithromycin and sotalol were co administered. The secondary objective was to correlate the QT interval changes induced by the study drugs to plasma concentrations during the elimination phase. METHODS: Twenty-four women received sotalol (160 mg) together with placebo or telithromycin (800 mg) in a two-period, double-blind, randomized study. Electrocardiograms were recorded at rest. Comparison of maximal corrected QT interval (QTc(max)) with sotalol in the presence or absence of telithromycin was performed. The relation between sotalol concentration and QTc was studied using linear regression. RESULTS: Mean difference (95% CI) between QTc(max) with sotalol-placebo and QTc(max) with sotalol-telithromycin was -15.5 ms (-27.7 to 3.2 ms). QTc(max) interval prolongation was lower (P < 0.05) with sotalol telithromycin than with sotalol-placebo, in relation to decreased sotalol plasma concentrations. Regression analysis showed that the relationship between sotalol plasma concentration and QTc interval duration was not modified by telithromycin co-administration. CONCLUSION: Our results do not support a potential synergistic effect on QT interval prolongation between sotalol and telithromycin. The decrease of mean QTc interval in subjects taking telithromycin and sotalol may be explained by a decrease of sotalol concentration. PMID- 16042665 TI - The central nervous system effects, pharmacokinetics and safety of the NAALADase inhibitor GPI 5693. AB - AIM: The aim was to assess the central nervous system (CNS) effects, pharmacokinetics and safety of GPI 5693, an inhibitor of a novel CNS-drug target, NAALADase which is being evaluated for the treatment of neuropathic pain. METHODS: This was a double-blind, placebo-controlled, exploratory study in healthy subjects receiving oral GPI 5693 single ascending doses of 100, 300, 750, 1125 mg with a placebo treatment randomly interspersed. An open-label, parallel extension examined the effects of food and sex on the pharmacokinetics of 750, 1125 and 1500 mg doses. Blood samples were collected for pharmacokinetic and biochemical/haematological safety analysis, vital signs, ECG and adverse event checks were performed regularly up to 48 h postdose. Postdose CNS effects were assessed using eye movements, adaptive tracking, electroencephalography (EEG), body sway and Visual Analogue Scales (VAS). RESULTS: CNS effects were mainly observed after the 1125 mg dose, showing a significant decrease of adaptive tracking performance, VAS alertness and VAS mood, and an increase of EEG occipital alpha and theta power. Gastro-intestinal (GI) adverse effects were frequent at higher doses. No clinically significant changes in vital signs or ECG were noted during any of the treatments. The therapeutically relevant concentration range (950-11 100 ng ml(-1)) as determined from animal experiments was already reached after the 300 mg dose. C(max) after the 300 mg and 750 mg dose was 2868 and 9266 ng ml(-1) with a t(1/2) of 2.54 and 4.78 h, respectively. Concomitant food intake (with the 750 mg and 1125 mg doses) reduced C(max) by approximately 66% and AUC by approximately 40%. With concomitant food intake, the dose-normalized C(max) also decreased significantly by -5.6 (CI: -2.6 to -8.7) ng ml(-1) mg(-1). The pharmacokinetic variability was largest after the 300 mg and 750 mg dose, resulting in a SD of approximately 50% of the C(max). CONCLUSION: NAALADase inhibition with GPI 5693 was safe and tolerable in healthy subjects. Plasma concentrations that were effective in the reversal of hyperalgesia in the chronic constrictive injury animal model of neuropathic pain were obtained at doses of 300, 750 and 1125 mg in the fasted state. Comcomitant food intake reduced C(max) and AUC. CNS effects and GI AEs increased in incidence over placebo only at the 1125 mg dose. PMID- 16042666 TI - Inappropriate drug prescribing in older adults: the updated 2002 Beers criteria- a population-based cohort study. AB - BACKGROUND: In 1997, Beers and colleagues developed explicit criteria for potentially inappropriate drug prescribing in ambulatory older adults aged 65 years and over. Several studies have been performed to estimate the prevalence of inappropriate drug prescribing based on these criteria. In 2002, the criteria were updated. AIMS: To examine the extent and trend of inappropriate drug prescribing to ambulatory older adults in the Netherlands between 1997 and 2001, according to the 1997 and the updated Beers criteria. METHODS: Data were retrieved from the Integrated Primary Care Information (IPCI) project, a general practice research database with data from computer-based patient records of a group of 150 general practitioners in the Netherlands. All subjects aged 65 and over were included. Prescriptions were classified as inappropriate if they fulfilled the Beers criteria of prescriptions that generally should be avoided in older adults because of a high risk of adverse effects, while also considering dose and comorbidity. RESULTS: Between 1997 and 2001, the 1-year risk of receiving at least one inappropriate drug prescription for older adults ranged between 16.8% (95% CI: 16.3-17.3%) and 18.5% (18.3-18.7%) according to the 1997 criteria and between 19.1% (18.6-19.6%) and 20.0% (19.5-20.5%) according to the updated Beers criteria. According to the updated criteria, the most frequently prescribed inappropriate drugs were nitrofurantoin, long-acting benzodiazepines, amitriptyline, promethazine and cimetidine. Temazepam and zolpidem were mostly prescribed in supratherapeutic dose. Conventional NSAIDs in persons with a history of gastric/duodenal ulcer were the most frequently prescribed contra indicated drugs. CONCLUSIONS: Prescribing potentially inappropriate prescriptions to ambulatory older people in the Netherlands is substantial. Compared with other studies using the 1997 Beers criteria, inappropriate prescribing to the elderly is lower than in the USA but similar to Finland. Despite a restrictive medication policy and a growing attention for medication surveillance in Europe, inappropriate drug prescribing is still a substantial problem. PMID- 16042667 TI - Off-label prescribing to children: attitudes and experience of general practitioners. AB - AIM: To identify experience with and attitudes towards paediatric off-label prescribing in primary care. METHOD: A prospective questionnaire survey was sent to a sample of Scottish primary care practices (346 doctors in 80 general practices located throughout Scotland). RESULTS: Two hundred and two (58%) completed questionnaires were returned. Over 70% of GPs admitted to being familiar with the concept, and 40% to knowingly prescribing off-label. The most important sources of paediatric prescribing information were the British National Formulary (81%), personal experience (71%) and previous prescription notes (45%). The most common reason given by GPs for off-label prescribing was prescribing for a younger age than recommended, although prescribing data confirm that age is the least important and dose the most important reason for such prescribing. When asked to comment upon different causes for off-label prescribing, 80% of respondents expressed appropriate awareness of and concern for the described scenarios. Over 97% of GPs ranked development of paediatric formulations and clearer dosage information more highly than clinical trials as a means to reducing off-label prescribing. CONCLUSIONS: Despite high levels of off-label prescribing in primary care in the UK, the majority of GPs claimed to be familiar with the concept, although less than half were aware of this common practice. A clear disparity between perceived and actual reasons for off-label prescribing was noted, possibly due to a reliance on personal experience, colleague experience or previous patient prescription notes as a guide to prescribing. PMID- 16042668 TI - Statin use after acute myocardial infarction: a nationwide study in Denmark. AB - AIMS: To study outpatient statin use after first acute myocardial infarction (AMI) in Denmark between 1995 and 2002 and to determine the predictors of statin use. METHODS: This is a nationwide population-based study using administrative registries. Patients with first AMI between 1995 and 2002 older than 30 years of age and alive 6 months after discharge (n = 45 219) were identified through the National Patient Registry. The statins purchased by these patients within 6 months after discharge were determined using the Registry of Medicinal Product Statistics, a nationwide prescription database. RESULTS: Statin use following AMI increased from 13% in 1995 to 61% in 2002. In 2002, 81% of patients aged 30-64 years used statins. Older patients used fewer statins, but use increased more among patients aged 75-84 years: from 1.3% to 43%. Use in elderly patients did not differ according to gender in 2000-02, but young men used more than younger women. In 2000-02, patients with diabetes (odds ratio (OR): 0.84; 95% confidence interval (CI): 0.74-0.95) and with heart failure (OR: 0.70; 95% CI: 0.64-0.76) were undertreated; this trend was present throughout the period. CONCLUSIONS: In this nationwide study, younger patients after AMI had high statin use in 2002, but high-risk patients such as those with diabetes and heart failure were still being undertreated. PMID- 16042669 TI - Meta-analysis of the influence of MDR1 C3435T polymorphism on digoxin pharmacokinetics and MDR1 gene expression. AB - AIMS: Studies revealing conflicting results of the functional significance of MDR1 exon 26 C3435T SNP on the disposition of digoxin in different ethnic groups led us to perform a meta-analysis on published data investigating the influence of C3435T SNP on the pharmacokinetics of digoxin and the expression of MDR1. METHODS: Meta-analysis was performed on data from published studies investigating the influence of MDR1 C3435T SNP on digoxin pharmacokinetics, as well as MDR1 expression in Caucasian and Japanese populations. The following outcomes were included: exposures to digoxin measured by area under the concentration-time curve and maximum concentration, the mean intestinal MDR1 mRNA expression and P gp expression in the absence of digoxin administration. RESULTS: The overall results of the meta-analysis in Caucasian and Japanese subjects suggested no major influence of the C3435T SNP on exposure levels of digoxin as determined by AUC(0-4 h) or AUC(0-24 h) although C(max) values for digoxin were lower in wild type (CC) subjects compared with subjects harbouring TT genotypes. Subgroup analysis by ethnic populations showed the oral availability of digoxin to be lower in wild-type Caucasian populations compared with wild-type Japanese subjects. No causal relationships were detected between the C3435T SNP and MDR1 mRNA or protein expression. CONCLUSIONS: Our meta-analysis of available studies indicates that the synonymous MDR1 C3435T SNP does not affect the pharmacokinetics of digoxin and the expression of MDR1 mRNA. Future studies should focus on the impact of MDR1 haplotypes on the pharmacokinetics of MDR1 substrates rather than the C3435T SNP alone. PMID- 16042670 TI - Inhibition of the metabolism of brotizolam by erythromycin in humans: in vivo evidence for the involvement of CYP3A4 in brotizolam metabolism. AB - AIMS: To obtain in vivo evidence for the involvement of cytochrome P450 (CYP) 3A4 in the metabolism of brotizolam. METHODS: Fourteen healthy male volunteers received erythromycin 1200 mg day(-1) or placebo for 7 days in a double-blind randomized crossover manner. On the 6th day they received a single oral 0.5-mg dose of brotizolam, and blood samplings were performed for 24 h. RESULTS: Erythromycin treatment significantly increased the peak plasma concentration (P < 0.05), total area under the plasma concentration-time curve (P < 0.01), and elimination half-life (P < 0.01) of brotizolam. CONCLUSIONS: The present study provides in vivo evidence for the involvement of CYP3A4 in brotizolam metabolism. PMID- 16042671 TI - Kidneys contribute to the extrahepatic clearance of propofol in humans, but not lungs and brain. AB - AIMS: The principal site for the metabolism of propofol is the liver. However, the total body clearance of propofol is greater than the generally accepted hepatic blood flow. In this study, we determined the elimination of propofol in the liver, lungs, brain and kidneys by measuring the arterial-venous blood concentration at steady state in patients undergoing cardiac surgery. METHODS: After induction of anaesthesia, propofol was infused continuously during surgery. For measurement of propofol concentration, blood samples were collected from the radial and pulmonary artery at predetermined intervals. In addition, blood samples from hepatic and internal jugular vein were collected at the same times in 19 patients in whom a hepatic venous catheter was fitted and the other six in whom an internal jugular venous catheter was fitted, respectively. In six out of 19 patients fitted with a hepatic venous catheter, blood samples from the radial artery and the renal vein were also collected at the same time, when the catheter was inserted into the right renal vein before insertion into the hepatic vein. RESULTS: Hepatic clearance of propofol was approximately 60% of total body clearance. The hepatic extraction ratio of propofol was 0.87 +/- 0.09. There was no significant difference in the concentration of propofol between the radial, pulmonary arteries and internal jugular vein. However, a high level of propofol extraction in the kidneys was observed--the renal extraction ratio being 0.70 +/- 0.13. CONCLUSIONS: We have demonstrated substantial renal extraction of propofol in human. Metabolic clearance of propofol by the kidneys accounts for almost one third of total body clearance and may be the major contributor to the extrahepatic elimination of this drug. PMID- 16042672 TI - Pharmaceutical care of patients with heart failure. AB - AIM: The aim of this study was to investigate the impact of a pharmacist-led pharmaceutical care programme, involving optimization of drug treatment and intensive education and self-monitoring of patients with heart failure (HF) within the United Arab Emirates (UAE), on a range of clinical and humanistic outcome measures. METHODS: The study was a randomized, controlled, longitudinal, prospective clinical trial at Al-Ain Hospital, Al-Ain, UAE. Patients were recruited from the general medical wards and from cardiology and medical outpatient clinics. HF patients who fulfilled the entrance criteria, and had no exclusion criteria present, were identified for inclusion in the study. After recruitment, patients were randomly assigned to one of two groups: intervention group or control group. Intervention patients received a structured pharmaceutical care service while control patients received traditional services. Patient follow-up took place when patients attended scheduled outpatient clinics (every 3 months). A total of 104 patients in each group completed the trial (12 months). The patients were generally suffering from mild to moderate HF (NYHA Class 1, 29.5%; Class 2, 50.5%; Class 3, 16%; and Class 4, 4%). RESULTS: Over the study period, intervention patients showed significant (P < 0.05) improvements in a range of summary outcome measures [AUC (95% confidence limits)] including exercise tolerance [2-min walk test: 1607.2 (1474.9, 1739.5) m.month in intervention patients vs. 1403.3 (1256.5, 1549.8) in control patients], forced vital capacity [31.6 (30.8, 32.4) l.month in the intervention patients vs. 27.8 (26.8, 28.9) in control patients], health-related quality of life, as measured by the Minnesota living with heart failure questionnaire [463.5 (433.2, 493.9) unit.month in intervention patients vs. 637.5 (597.2, 677.7) in control patients; a lower score in this measure indicates better health-related quality of life]. The number of individual patients who reported adherence to prescribed medications was higher (P < 0.05) in the intervention group (85 vs. 35), as was adherence to lifestyle advice (75 vs. 29) at the final assessment (12 months). There was a tendency to have a higher incidence of casualty department visits by intervention patients, but a lower rate of hospitalization. CONCLUSIONS: The research provides clear evidence that the delivery of pharmaceutical care to patients with HF can lead to significant clinical and humanistic benefits. PMID- 16042673 TI - Do nonsteroidal anti-inflammatory drugs affect the risk of developing ovarian cancer? A meta-analysis. AB - AIM: The relationship between the use of nonsteroidal anti-inflammatory drugs (NSAIDs), including aspirin, and the risk of ovarian cancer has been controversial. This study examines the strength of this association by conducting a detailed meta-analysis of the studies published in peer-reviewed literature on the subject. METHODS: A comprehensive search for articles published up to April 2004 was performed, reviews of each study were conducted and data were abstracted. Prior to meta-analysis, the studies were evaluated for publication bias and heterogeneity. Pooled relative risk estimates (RR) and 95% confidence intervals (CIs) were calculated. RESULTS: Ten reports (six case-control and four cohort studies), published between 1998 and 2004, were identified. There was no evidence of an association between aspirin use and ovarian cancer risk either assuming a random-effects model (RR = 0.92, 95% CI 0.80, 1.06), or a fixed effects model (RR = 0.93, 95% CI 0.81, 1.06). Similarly, we did not find evidence of an association between non-aspirin NSAID use and ovarian cancer, both on the basis of a random-effects model (RR = 0.86, 95% CI 0.68, 1.08), and on the basis of a fixed-effects model (RR = 0.88, 95% CI 0.76, 1.01). When the analyses were stratified into subgroups, there was no evidence that study design substantially influenced the estimate of effects. Furthermore, our analysis did not show decreasing risks with increasing frequency or duration of use, features often associated with causal relationships. CONCLUSIONS: Our meta-analysis findings do not support that NSAID use plays a role in the chemoprevention of ovarian cancer. Future research should examine potential relationships between specific NSAIDs and ovarian cancer, taking into account the possible biases that may have affected this meta-analysis. PMID- 16042674 TI - Angiotensin II receptor blocker-induces blunted taste sensitivity: comparison of candesartan and valsartan. AB - AIMS: To compare the degree of taste disturbance by candesartan and valsartan. METHODS: Candesartan cilexetil (4 mg day(-1)), valsartan (40 mg day(-1)), or vehicle was given to subjects (n = 8) for 13 days in a randomized, placebo controlled, three-way crossover design with a 14-days washout period. Gustometry by filter-paper test and electrogustometry were performed before and at the end of each trial. Plasma renin activity and zinc concentrations in serum and saliva were measured. RESULTS: Detection thresholds of four basic tastes (sweet, salty, sour and bitter) by paper-disc test and electrogustometry were significantly worsened and plasma renin activity was elevated after the test, while the effects of two drugs did not significantly differ. These drugs did not affect zinc concentrations. CONCLUSION: Both candesartan and valsartan similarly alter taste sensitivity after the repeated dosing of the drug. PMID- 16042675 TI - Coadministration of gemfibrozil and itraconazole has only a minor effect on the pharmacokinetics of the CYP2C9 and CYP3A4 substrate nateglinide. AB - BACKGROUND AND AIMS: Gemfibrozil, and particularly its combination with itraconazole, greatly increases the area under the plasma concentration-time curve [AUC(0, infinity)] and response to the cytochrome P450 (CYP) 2C8 and 3A4 substrate repaglinide. In vitro, gemfibrozil is a more potent inhibitor of CYP2C9 than of CYP2C8. Our aim was to investigate the effects of the gemfibrozil itraconazole combination on the pharmacokinetics and pharmacodynamics of another meglitinide analogue, nateglinide, which is metabolized by CYP2C9 and CYP3A4. METHODS: In a randomized crossover study with two phases, nine healthy subjects took 600 mg gemfibrozil and 100 mg itraconazole (first dose 200 mg) twice daily or placebo for 3 days. On day 3, they ingested a single 30-mg dose of nateglinide. Plasma nateglinide and blood glucose concentrations were measured for up to 12 h. RESULTS: During the gemfibrozil-itraconazole phase, the AUC(0, infinity) and C(max) of nateglinide were 47% (range 23-74%; P < 0.0001) and 30% (range - 8% to 104%; P = 0.0146) higher than during the placebo phase, respectively, but the t(max) and t1/2 of nateglinide remained unchanged. The combination of gemfibrozil and itraconazole had no effect on the formation of the M7 metabolite of nateglinide but impaired its elimination. The blood glucose response to nateglinide was not significantly changed by coadministration of gemfibrozil and itraconazole. CONCLUSIONS: The combination of gemfibrozil and itraconazole has only a limited influence on the pharmacokinetics of nateglinide. This is in marked contrast to the substantial effect of this combination on the pharmacokinetics of repaglinide. The findings suggest that in vivo gemfibrozil, probably due to its metabolites, is a much more potent inhibitor of CYP2C8 than of CYP2C9. PMID- 16042676 TI - Profound weight loss associated with reboxetine use in a 44-year-old woman. AB - We report a case of significant weight loss experienced by a 44-year-old Caucasian woman treated with reboxetine. She was treated with this drug at 12 mg daily for a total duration of 11 months. During the corresponding period her body mass index (BMI) decreased from a baseline of 21.4 kg m(-2) to a low of 16.8 kg m(-2). Withdrawal of the drug led to a full recovery of her BMI. The strongest evidence linking reboxetine to this woman's weight loss laid in the fact that the re-introduction of the drug subsequently caused a similar negative impact in her BMI. PMID- 16042677 TI - A distance-learning programme in pharmacovigilance linked to educational credits is associated with improved reporting of suspected adverse drug reactions via the UK yellow card scheme. AB - AIMS: The effect of a distance-learning package linked to educational credits on the rate and quality of spontaneous adverse drug reaction (ADR) reporting by general practitioners (GPs) and pharmacists in Wales was investigated. METHODS: In April 2000, 477 GPs and 261 pharmacists enrolled in the 12 month programme. RESULTS: The number and quality of yellow card reports improved compared with those of a control region in England (Northern Region). CONCLUSIONS: We conclude that an educational initiative in drug safety linked to incentives may be associated with a significant but perhaps short-lived improvement in the rate and quality of ADR reporting. PMID- 16042678 TI - Effect of pregnancy on a measure of FMO3 activity. PMID- 16042680 TI - Lymphoma and malignant vasovagal syndrome. PMID- 16042681 TI - Much ado about nothing: silent brain lesions in T-ALL. PMID- 16042682 TI - Diagnostic criteria for monoclonal B-cell lymphocytosis. AB - Very low levels of circulating monoclonal B-cell subpopulations can now be detected in apparently healthy individuals using flow cytometry. We propose the term 'monoclonal B-cell lymphocytosis' (MBL) to describe this finding. The aim of this document is to provide a working definition of MBL for future clinical, epidemiological and laboratory studies. We propose that the detection of a monoclonal B-cell population by light chain restriction is sufficient to define this condition in individuals not meeting the diagnostic criteria for other B lymphoproliferative disorders. The majority of individuals with MBL will have cells that are indistinguishable from chronic lymphocytic leukaemia (CLL). However, this blood cell clonal expansion of CD5+ or CD5- B-lymphocytes is age dependent and immunophenotypic heterogeneity is common. Longitudinal studies are required to determine whether MBL is a precursor state to CLL or other B lymphoproliferative disease in a situation analogous to a monoclonal gammopathy of undetermined significance and myeloma. Future studies of MBL should be directed towards determining its relationship to clinical disease, particularly in individuals from families with a genetic predisposition to developing CLL. PMID- 16042683 TI - Perinatal stroke--risk factors and management. AB - Stroke is an uncommon but increasingly recognised cause of mortality and long term neurological morbidity in children. A significant number of these events appear to be caused by thromboembolic disease and, as with other childhood thrombotic problems, the incidence of central nervous system events appears highest during the neonatal period. In contrast to peripheral arterial and venous thrombotic problems, it is likely that a proportion of cerebral thromboembolic events occur either in utero or perinatally and reflect different risk factors from those occurring in older infants and children. The pathophysiology of perinatal stroke is complex and in many cases is likely to be multifactorial. It is now recognised that risk factors may relate to both maternal and placental problems as well as fetal and neonatal disorders. Large prospective studies of perinatal stroke are currently lacking and efforts to define the relative contribution from each of these areas are at an early stage. The complex nature of these disorders requires collaboration between a number of different disciplines including obstetrics, fetal medicine, pathology, neonatology and neurology. Of particular interest to haematologists is the possible impact of prothrombotic abnormalities in the pathophysiology of these events and also the potential for the use of antithrombotic agents in both management and prevention. PMID- 16042684 TI - Immunotherapy in multiple myeloma--possibility or probability? AB - In a small number of patients with multiple myeloma (MM), long-term disease-free survival has been achieved by harnessing the immune phenomenon, 'graft-versus tumour' effect, induced by allogeneic haemopoietic stem cell transplantation. This has prompted many investigators to examine ways in which a patient's own immune system can be more effectively directed against their disease, with the ultimate aim of tumour eradication. In this review we assess the current understanding of immunobiology in MM, and how the different components of the immune system, such as dendritic cells, T cells and natural killer cells, may be harnessed using in-vitro and in-vivo priming techniques. We look at the clinical immunotherapy trials reported to date and whether, in light of the current information, immunotherapy for MM is an achievable goal. PMID- 16042685 TI - Conventional second-line salvage chemotherapy regimens are not warranted in patients with malignant lymphomas who have progressive disease after first-line salvage therapy regimens. AB - This study aimed to determine the outcome of patients with relapsed or refractory lymphoma who have an inadequate response to first-line salvage therapy (1 degrees ST) and who subsequently receive a second-line salvage regimen (2 degrees ST) with the intention of ultimately proceeding to high-dose therapy. The outcome of 57 patients [Hodgkin's Lymphoma 17, histologically-aggressive non-Hodgkin's Lymphoma (NHL) 26, histologically-indolent NHL 14] who received more than one modality of conventional-dose salvage therapy was analysed. Sixteen patients had a partial response (PR) to 1 degrees ST, but subsequently received 2 degrees ST because the PR was judged to be inadequate (iPR) because of persisting disease bulk or marrow infiltration. Of these 16 patients, 10 (63%) continued to respond to 2 degrees ST. Of the 15 patients who had stable disease following 1 degrees ST, 5 (33%) responded to 2 degrees ST. Only one of the 24 (4%) with progressive disease (PD) following 1 degrees ST, responded to 2 degrees ST. 25 of the 57 patients ultimately underwent stem cell transplantation. The 2-year progression free survival (PFS) and the 3-year overall survival (OS) for all patients was 24% and 31%, respectively. Long-term survival was highly dependent on response to 1 degrees ST (P = 0.0001); in patients with PD following 1 degrees ST, the PFS and OS at 3 years was only 4%. This analysis indicates that patients with malignant lymphomas, who have PD on 1 degrees ST, are not rescued by subsequent salvage regimens. They should either be treated palliatively or novel approaches should be explored. PMID- 16042686 TI - Effects of imatinib and interferon on primitive chronic myeloid leukaemia progenitors. AB - Imatinib has impressive activity against chronic myeloid leukaemia (CML), but does not appear to completely eradicate the disease. Although responses to interferon-alpha (IFN) are slower and less dramatic than those to imatinib, they can be durable even after discontinuation of the drug. Unlike imatinib, the specific mechanisms responsible for IFN's clinical activity in CML are unknown. We found that IFN induced a G1 cell cycle arrest, as well as terminal differentiation, of the CML cell line KT-1 and CML CD34+ cells from clinical specimens. Myeloid growth factors augmented the antileukaemic activity of IFN, and neutralising antibodies directed against myeloid growth factors inhibited IFN's antileukaemic activity. We next directly compared the effects of imatinib and IFN against differentiated and primitive CML progenitors from newly-diagnosed patients. Although less active against CML granulocyte-macrophage colony forming units than imatinib, IFN was significantly more toxic to primitive CML progenitors responsible for the maintenance of long-term cultures. Imatinib and IFN appear to have divergent effects on CML progenitors at different stages of maturation, with imatinib more active against differentiated CML progenitors and IFN more active against primitive CML progenitors. The different target cells for these agents may explain the disparities in the kinetics and durability of their clinical responses. At least part of the clinical effect of IFN in CML appears to result from its ability to differentiate primitive CML progenitors. PMID- 16042687 TI - Serological identification of adult T-cell leukaemia-associated antigens. AB - Adult T-cell leukaemia (ATL) is a peripheral T-cell neoplasm caused by human T cell leukaemia virus type I (HTLV-I). Several clinical observations suggest that some tumour-associated antigens in ATL may be recognised by the immune system. In this study, we performed the serological screening of an expression library to identify ATL-associated antigens by using materials from a unique ATL patient with long-term stable disease. Among five distinct genes isolated, serine/arginine protein kinase 1 (SRPK1), which has been reported to have a restricted normal tissue distribution, was found to be overexpressed in most acute type ATL samples, but not in chronic type ATL or in normal peripheral blood mononuclear cells by real-time reverse transcription polymerase chain reaction. Interestingly, the overexpression of SRPK1 in aggressive types of ATL was more exclusively observed at the protein level than at the mRNA level. Autologous antibody to SRPK1 was confirmed in the ATL patient using Western blot analysis with plasma, but not detected in asymptomatic HTLV-I carriers or in healthy volunteers. These results indicate that SRPK1 may be useful for the development of therapeutic and diagnostic methods for patients with ATL. PMID- 16042688 TI - Clonal granulocytes in polycythaemia vera and essential thrombocythaemia have shortened telomeres. AB - Summary The purpose of this study was to evaluate telomere length in peripheral blood granulocytes and mononuclear cells collected from 22 women with polycythaemia vera (PV) and essential thrombocythaemia (ET). PV and ET are chronic myeloproliferative diseases whose heterogeneity of stem cell origin and clonal development has been established through analysis of X-chromosome inactivation patterns. The results from clonality assay and determination of telomere length show that only clonal granulocytes have shortened telomeres. PMID- 16042689 TI - Influence of the intensity of the conditioning regimen on the characteristics of acute and chronic graft-versus-host disease after allogeneic transplantation. AB - The graft-versus-host disease (GVHD) characteristics of 150 consecutive patients undergoing reduced intensity conditioning allogeneic (allo-RIC) transplants and 88 patients undergoing myeloablative conditioning regimen were analysed. All patients received the same GVHD prophylaxis and peripheral blood stem cells from a human leucocyte antigen identical sibling. The cumulative incidence of acute GVHD (aGVHD) was 67% and 44% in the myeloablative and allo-RIC regimen groups, respectively (P < 0.001), and was 39% vs. 29%, respectively (P = 0.043), for grades 2-4 aGVHD. Only conditioning type (myeloablative versus allo-RIC) significantly influenced the incidence of aGVHD in multivariate analysis: Hazard ratio (HR) = 2.16 [95% confidence interval (CI): 1.52-3.07], P < 0.0001. The cumulative incidence of chronic GVHD (cGVHD) was 63% and 71% among myeloablative and allo-RIC patients respectively (P = 0.084). This trend was because of the higher incidence of limited cGVHD, but not extensive cGVHD among allo-RIC recipients [HR = 3.3 (95% CI: 1.42-8.08), P = 0.0017]. Moreover, among patients who developed cGVHD, the cumulative incidence of limited cGVHD was significantly lower in the myeloablative group than in the allo-RIC group (7% vs. 25%, P = 0.007). Duration of immunosuppression was shorter among allo-RIC patients (35.5% vs. 68.8% required systemic immunosuppression 36 months after transplant, P = 0.028). Although prospective controlled trials are required to further evaluate the effect of the conditioning regimen on GVHD, our results suggest that RIC modifies the incidence and characteristics of both acute and cGVHD after allogeneic transplantation, and decreases the immunosuppression requirements in long-term follow up when compared with myeloablative conditioning. PMID- 16042690 TI - Transient familial haemophagocytic lymphohistiocytosis reactivation post-CD34 haematopoietic stem cell transplantation. AB - Familial haemophagocytic lymphohistiocytosis (FHLH) is a genetic disorder caused by defective lymphocyte cytotoxicity, resulting in impaired lymphocyte homeostasis and macrophage infiltration of solid tissues and bone marrow, with extensive haemophagocytosis. It is invariably fatal unless treated by allogeneic haematopoietic stem cell transplantation (HSCT). In a retrospective analysis of 11 cases of FHLH, transplanted in one centre between January 1999 and December 2003, it was found that host T cell expansion occurred early after HSCT in a setting of a viral infection (cytomegalovirus and Epstein-Barr virus respectively) in two cases who received T cell-depleted HSCT. Transient recurrence of clinical and biological manifestations of FHLH was observed, despite evidence for donor cell engraftment. Secondary development of donor T cells led to stable mixed chimaerism and sustained remission of FHLH. Detection of host-derived T cells soon after HSCT in a patient with FHLH should thus not mistakenly be taken as a manifestation of graft rejection. PMID- 16042691 TI - Sirolimus in combination with tacrolimus and corticosteroids for the treatment of resistant chronic graft-versus-host disease. AB - Chronic graft-versus-host disease (cGVHD) remains a major cause of morbidity and mortality in haematopoietic transplant recipients. Sirolimus is a macrocyclic triene antibiotic with immunosuppressive, antifungal and antitumour properties, that has activity in the prevention and treatment of acute GVHD. We conducted a phase II trial of sirolimus combined with tacrolimus and methylprednisolone in patients with steroid-resistant cGVHD. Thirty-five patients who developed GVHD after day 100 post-transplant were studied. Six patients had a complete response and 16 a partial response with an overall response rate of 63%. Major adverse events related to the combination of tacrolimus and sirolimus were hyperlipidaemia, renal dysfunction and cytopenias. Four patients had thrombotic microangiopathy (TMA) and 27 (77%) had infectious complications. The median survival for the whole group was 15 months. A significantly better outcome was observed in patients with a platelet count > or = 100 x 10(9)/l, as well as in those with true chronic manifestations of GVHD compared to those with acute GVHD beyond day 100. Controlled trials comparing this approach with alternative strategies to determine which can best achieve the goal of GVHD-free survival are warranted. PMID- 16042692 TI - Identification of APOBEC3B as a potential target for the graft-versus-lymphoma effect by SEREX in a patient with mantle cell lymphoma. AB - Recently, a graft-versus-lymphoma (GVL) effect has been shown in patients allografted for mantle cell lymphoma (MCL), but the antigenic targets of this response remain unclear. We screened an MCL cDNA expression library with sera at GVL response after allogeneic haematopoietic stem cell transplantation and isolated five genes including APOBEC3B. Antibody responses to APOBEC3B were correlated with clinical response. Furthermore, APOBEC3B-specific T-cell response was induced with one of the HLA-A*0201 binding peptides. In addition, APOBEC3B mRNA appeared to be upregulated in some MCL. These findings warrant further investigation of APOBEC3B as a candidate gene eliciting an effective immune response against MCL. PMID- 16042693 TI - Environmental risk factors for inhibitor development in children with haemophilia A: a case-control study. AB - This case-control study investigated the interactions between genetic and environmental factors and inhibitor development in 108 children with haemophilia A exclusively treated with recombinant factor VIII (FVIII). Sixty patients with inhibitors were compared with 48 inhibitor-free controls. Family history of inhibitors and null mutations in the FVIII gene were more prevalent in cases than in controls (20% vs. 2%, P = 0.001 and 83% vs. 64%, P = 0.04, respectively). On the other hand, there was no difference between cases and controls for such putative risk factors of inhibitor development as amniocentesis/villocentesis, premature/caesarean birth, breast-feeding, treatment during infections/vaccinations, surgical procedures and central nervous system bleeding. A trend was found for an increased risk of inhibitor development in children first treated at a young age (<11 months); however, this was not confirmed after adjusting for genetic factors. The implementation of prophylaxis was evaluated as a putative risk factor in a subgroup of 25 cases: seven who started prophylaxis prior to inhibitor development and 18 potentially eligible for prophylaxis because they were inhibitor-free up to the age of 35 months (i.e. the upper limit of the age range at prophylaxis onset in cases and the median age at prophylaxis onset in controls). Patients who started prophylaxis had a lower inhibitor risk than those treated on demand (adjusted odds ratio 0.2, 95% confidence interval 0.06-0.9). The protective effect on inhibitor development shown by prophylaxis may represent an additional advantage prompting its use in haemophilic children. PMID- 16042694 TI - AML1B transcriptional repressor function is impaired by the Flt3-internal tandem duplication. AB - Fms-like tyrosine kinase 3 (Flt3) is a type III receptor tyrosine kinase. The internal tandem duplication (ITD) of the juxtamembrane region of this receptor is the most prevalent mutation in acute myeloid leukaemia (AML). The silencing mediator of retinoic and thyroid hormone receptors (SMRT) co-repressor recruits histone deacetylases (HDAC) and mediates transcriptional repression by interacting with various transcription factors. We recently reported that Flt3 ITD interferes with the transcriptional and biological action of promyelocytic leukaemia zinc finger transcriptional repressor by dissociating it from SMRT. In this study, we aimed to clarify whether the repressional activity of other well known oncoproteins, such as AML1/Runx1 (AML1), is also affected by Flt3-ITD. We verified that the repression activity of AML1B, the isoform of AML1, is dependent on HDAC activity by using HDAC inbitor trichostatin A in GAL4 reporter assays. Mammalian two-hybrid assays demonstrated that this protein interacts with SMRT. Furthermore, this AML1B-SMRT interaction was disrupted by the overexpression of Flt3-ITD, leading to the reduction of AML1B repression activity. Additionally, we showed AML1B repression target, p21 (WAF1/CIP1), was aberrantly expressed in Flt3 ITD stably expressed BaF3 cells. Taken together, Flt3-ITD disrupts transcriptional repressor functions resulting in aberrant gene regulation in leukaemic cells. PMID- 16042695 TI - Health-related quality of life in children with sickle cell disease: child and parent perception. AB - Health-related quality of life (HRQL) is an outcome that may be used to measure the impact of sickle cell disease on the child and their family but has not been routinely assessed in this disease. The objective of this study was to describe the HRQL of children with sickle cell disease as reported by the parent and the child, to compare the relationship between the two, and to determine the association of parent, child and disease characteristics on HRQL. Ninety-five parents completed the Child Health Questionnaire (CHQ)-Parent Form28 and 53 children completed the CHQ-Child Form87. Compared with the child report, parents reported worse HRQL in the overall perception of health, physical functioning, behaviour and self esteem domains of HRQL (P < 0.005). Parent and child reports of HRQL correlated strongly in assessment of the impact of bodily pain (r = 0.58) on HRQL and moderately in physical functioning (r = 0.44), behaviour (r = 0.45), general health (r = 0.44), self esteem (r = 0.40) and changes in health (r = 0.33) domains. Disease status, neurobehavioral co-morbidities, and parent education were associated with the HRQL of the child. Both the parent and child perspectives are needed to fully understand the impact of sickle cell disease on the HRQL of the child and effect of this disease on the family. PMID- 16042696 TI - Sildenafil therapy in patients with sickle cell disease and pulmonary hypertension. AB - Pulmonary hypertension is a frequent complication of sickle cell disease that is associated with haemolysis, impaired nitric oxide bioavailability and high mortality. We sought to evaluate the safety and efficacy of selective pulmonary vasodilators and antiproliferative agents in this at-risk population. After optimising sickle cell disease therapy to stabilise haemoglobin and fetal haemoglobin levels, we evaluated the safety and efficacy of sildenafil in 12 patients with sickle cell disease and pulmonary hypertension. Sildenafil therapy (mean duration 6 +/- 1 months) decreased the estimated pulmonary artery systolic pressure [50 +/- 4 to 41 +/- 3 mmHg; difference 9 mmHg, 95% confidence interval (CI): 0.3-17, P = 0.043] and increased the 6-min walk distance (384 +/- 30 to 462 +/- 28 m; difference 78 m, 95% CI: 40-117, P = 0.0012). Transient headaches occurred in two patients and transient eye-lid oedema in four patients. No episodes of priapism occurred in the three men in the study; two of them were on chronic exchange transfusions and one had erectile dysfunction. IN CONCLUSION: (1) sickle cell disease patients with anaemia and pulmonary hypertension have significant exercise limitation; (2) the 6-min walk distance may be a valid endpoint in this population; (3) therapy with sildenafil appears safe and improves pulmonary hypertension and exercise capacity. Additional phase I studies in males with sickle cell disease followed by phase II/III placebo controlled trials evaluating the safety and efficacy of sildenafil therapy in sickle cell disease patients with pulmonary hypertension are warranted. PMID- 16042697 TI - A novel 33.3 kb deletion (- -KOL) in the alpha-globin gene cluster: a brief report on deletional alpha-thalassaemia in the heterogeneous eastern Indian population. AB - We have detected, in three unrelated eastern Indian individuals, a hitherto unreported alpha zero deletion, - -KOL, in the heterozygous state, encompassing the embryonic zeta2-globin and the duplicated alpha-globin genes extending from c. 1150 bp upstream of the zeta2 globin gene to c. 960 bp downstream of the theta1 gene. Other deletions present in 120 unrelated, eastern Indian, putative alpha-thalassaemia patients are -3.7 kb (16.25%), -4.2 kb (5%) and - -SEA (3.33%). PMID- 16042698 TI - Different fatherhood as a possible explanation for difficult pedigrees in coagulation defects: only 'mater semper certa est'. PMID- 16042700 TI - Percentage of CD34 cells in a minimum 500-cell count in bone marrow trephines of patients with chronic myeloid leukaemia provides the best correlation with aspirate blast count. PMID- 16042701 TI - The Y/C1584 mutation of von Willebrand factor in type 2M von Willebrand disease: frequency and clearance of von Willebrand factor. PMID- 16042702 TI - Transfusion related acute lung injury (TRALI). PMID- 16042705 TI - How should we advise patients about the chemoprevention of prostate cancer? PMID- 16042706 TI - A proposal for a new classification for operative procedures for stress urinary incontinence. PMID- 16042707 TI - Renal transplantation and manpower issues. PMID- 16042708 TI - Is premature ejaculation all in the mind? PMID- 16042709 TI - Selecting therapy for maintaining sexual function in patients with benign prostatic hyperplasia. AB - In the first of these mini-reviews the selection of therapy for the maintenance of sexual function in patients with BPH is outlined, along with an explanation of how altered regulation of neurotransmitters, especially noradrenaline, may underlie the syndrome of LUTS and sexual dysfunction. Other mini-reviews outline the current status of robotic surgery to treat renal and adrenal disorders, and its future applications, and the potential use of the nitric oxide/cGMP pathway as a potential target to treat BOO associated with benign prostatic enlargement. Finally, the capacity to be creative in academic departments is extolled as a core property of academicians, and its surfacing described as having the potential to revitalize individuals and departments. PMID- 16042710 TI - Robotic renal and adrenal surgery: present and future. PMID- 16042711 TI - Targeting bladder outlet obstruction from benign prostatic enlargement via the nitric oxide/cGMP pathway? PMID- 16042712 TI - The vital role of creativity in academic departments. PMID- 16042713 TI - Phosphodiesterase type 5 inhibitors for erectile dysfunction. AB - The cyclic nucleotide signalling pathway mediates the smooth-muscle relaxing effects of nitric oxide necessary for normal erectile function. Down-regulation of this pathway is central to the pathophysiology of many forms of erectile dysfunction (ED), which is often associated with other chronic diseases (e.g. hypertension, type 2 diabetes mellitus) and treatments (e.g. certain drugs, radical prostatectomy). Conversely, selective inhibition of the enzyme that catalyses the degradation of cGMP (phosphodiesterase type 5, PDE-5) promotes erectile responses to sexual stimulation. The successful launch and commercialization of the selective PDE5 inhibitor (PDE5I) sildenafil transformed the treatment of ED, not only by providing an effective, well tolerated oral ED therapy, but also by fostering greater candour about the problem among men. Sildenafil is highly effective in promoting erectile responses across a wide spectrum of severity and causes of ED, including patients with ED that is often refractory to treatment. The recent advent of vardenafil, which has the highest in vitro potency of all available PDE5Is, and tadalafil, which has a prolonged half-life that may enable couples to have sexual activity with less planning, represent further advances. Other PDE5Is offering further potential improvements are under active investigation. PMID- 16042714 TI - Carbonic anhydrase IX and the future of molecular markers in renal cell carcinoma. AB - The use of carbonic anhydrase IX as a promising molecular marker in RCC is described by authors from Los Angeles, who discuss the promise that molecular markers hold to improve diagnosis, staging, treatment, surveillance and survival of patients with RCC. There is a whole range of new treatments being introduced in the management of metastatic renal cancer. The use of VEGF-targeted therapy has particular importance, especially as it has a strong genetically linked rationale for its potential success in this area. Authors from the USA show that substantial clinical activity has been reported in initial clinical trials. In prostate cancer, drugs targeting microtubules, such as taxanes, have already been introduced clinically, and their success has received widespread attention. A new group of drugs, the epothilones, have similar but not identical binding properties to microtubules, and authors from the USA describe how they have shown activity in hormone-refractory prostate cancer, and are moving to phase III testing. PMID- 16042715 TI - Therapy targeted at vascular endothelial growth factor in metastatic renal cell carcinoma: biology, clinical results and future development. PMID- 16042716 TI - Multidetector computed tomography vs magnetic resonance imaging for defining the upper limit of tumour thrombus in renal cell carcinoma: a study and review. AB - OBJECTIVE: To compare the findings of multidetector computed tomography (CT) with surgical pathology and magnetic resonance imaging (MRI), to determine the accuracy of delineating the superior extent of inferior vena cava (IVC) thrombotic involvement in renal cell cancer (RCC). PATIENTS AND METHODS: A prospective database was examined of 11 patients (median age 65 years, range 45 77) being assessed for suspected IVC extension of RCC tumour thrombus with both multidetector CT and MRI. All had pathology confirming RCC, and eight of those undergoing surgery had pathological confirmation of tumour thrombus extent. All images were analysed originally, then re-analysed by two independent radiologists, an experienced urologist and a urological trainee unaware of the original reports and other imaging results, with a final determination on tumour thrombus level by consensus. RESULTS: The multidetector CT results were completely accurate when compared with surgical specimens and were in agreement with MRI on all but one occasion, where MRI determined the renal vein to be clear when it was involved on CT and at surgery, giving MRI an accuracy of seven of eight samples. CONCLUSIONS: Whilst there were few patients and further studies are needed, multidetector CT was comparable with MRI in determining tumour thrombus level. More importantly, in the eight patients with surgical pathological confirmation, multidetector CT was accurate in all. Ultimately, it may replace MRI as the 'gold standard' for imaging to delineate the upper limit of tumour thrombosis in RCC. PMID- 16042717 TI - Epothilones and the next generation of phase III trials for prostate cancer. PMID- 16042718 TI - Tumour markers for managing men who present with metastatic prostate cancer and serum prostate-specific antigen levels of <10 ng/mL. AB - OBJECTIVE: To define immunohistochemical features of the primary cancers that might help in the differential diagnosis and monitoring of treatment in men presenting with metastatic prostate cancer and low serum levels of prostate specific antigen (PSA), who can be difficult to diagnose and manage. PATIENTS AND METHODS: Paraffin blocks of prostate biopsies were obtained for 33 patients presenting with untreated metastatic prostate cancer and serum PSA levels of <10 ng/mL. Sections were immunostained for PSA, prostatic acid phosphatase (PAP), prostate-specific membrane antigen (PSMA), androgen receptor (AR), chromogranin A and CD 56. RESULTS: The combined Gleason scores were 8-10 in 25 men (76%) and 6 or 7 in the other eight (24%). Morphologically, there were no neuroendocrine features. PSA immunostaining was equivocal in 12 (36%) cases and in a further 19 (58%) was strong but focal and could be missed on biopsy sampling. PSMA was expressed in 90% of cases, and staining was widely distributed in nine of the 12 in which PSA staining was equivocal. There was strong AR expression in 30 (91%) cases and it was present in areas where PSA was absent. CONCLUSION: In this patient group, immunohistochemical assessments of PSMA and AR are potentially useful as diagnostic markers. PMID- 16042719 TI - Age-specific reference levels of serum prostate-specific antigen and prostate volume in healthy Arab men. AB - OBJECTIVE; To determine age-specific reference ranges for serum prostate-specific antigen (PSA) concentration and prostate volumes in a population of healthy Arab men. SUBJECTS AND METHODS: Blood samples were taken from 396 healthy Arab men (from Kuwait and Oman) aged 15-79 years and from across the social spectrum. Men aged >40 years had a digital rectal examination and transrectal ultrasonography of the prostate to determine prostate volume. The serum PSA level was measured using commercial kits, and age-specific ranges for PSA levels and prostate volume determined. RESULTS: The serum PSA ranges (ng/mL) for each age range in Arab men were: 40-49 years, 0-0.9; 60-69, 0-2.7; 70-79, 0-5.5 ng/mL; the respective prostate volumes were 8-22, 9-30 and 10-33 mL. The serum PSA level and prostate volume correlated with age (P < 0.001). Arab men had lower serum PSA levels and prostate volumes than those reported for Caucasians, but similar to those reported for Asians (Japanese and Chinese). CONCLUSION: These results indicate that Arab men have lower PSA levels and prostate volumes than Caucasians. The levels are slightly lower than those reported in the Japanese and, as in the Japanese, low PSA levels and small prostate volumes might be related to the low incidence of clinical prostate cancer in Arab men. PMID- 16042720 TI - Analysis of peripheral blood for prostate cells after autologous transfusion given during radical prostatectomy. AB - OBJECTIVES: To determine if cells expressing prostate-specific antigen (PSA) can be detected in blood collected by a cell-saver during radical prostatectomy (RP) or in the peripheral blood after intraoperative autotransfusion (IAT). PATIENTS AND METHODS: In all, 112 men with clinical T1c-T2 prostate cancer undergoing RP were prospectively assessed. A cell-saver system was used in each to collect blood from the surgical field after prostate manipulation. IAT was given based on clinical indications. Standardized peripheral blood samples were collected from patients before RP, in the recovery room afterward, and at 3-5 weeks after surgery. A reverse-transcriptase-polymerase chain reaction assay for PSA mRNA was used to detect prostate cells in cell-saver and peripheral blood samples. Patients were followed after surgery with PSA measurements to assess biochemical failure. RESULTS: PSA-expressing cells were detected in 88% of cell-saver reservoir and 13% of preoperative blood samples. No PSA-expressing prostate cells were detected in any peripheral blood samples collected 3-5 weeks after surgery. Analysis of data with 40 months of follow-up showed IAT was not an independent predictor of biochemical failure in multivariate analysis. CONCLUSIONS: Although IAT blood contains PSA-expressing cells, none could be detected 3-5 weeks after surgery. IAT during RP was not associated with a greater risk of biochemical failure. PMID- 16042721 TI - New perioperative management reduces bleeding in radical retropubic prostatectomy. AB - OBJECTIVE: To describe the effect of modifications to radical retropubic prostatectomy (RRP, known to be associated with severe bleeding) on blood loss in a retrospective analysis comparing RRPs by one experienced surgeon before and after the changes. PATIENTS AND METHODS: The new method comprised reducing the intravenously applied volume, using a peridural catheter and maintaining a 25-30 degrees Trendelenburg position. The difference in haemoglobin before and after RRP was analysed before the changes (group 1) and after (group 2). If transfusions were required the haemoglobin value was corrected, whereby 1 mL of erythrocyte concentrate increased the patient's haemoglobin by 0.03 g/L. RESULTS: Assessment was possible in 201 of 234 patients, 110 from group 1 and 91 from group 2. The mean transfusion-corrected difference in haemoglobin was 53 g/L in group 1 (20% transfusion rate) and 35.2 g/L in group 2 (1.09% transfusion rate; P > 0.001). The median intravenous volume applied was 5.96 L in group 1 and 3.49 L in group 2 (P < 0.001). The complication rate did not differ between the groups. CONCLUSION: This new method minimizes the intraoperative blood loss during RRP; transfusions are only necessary in rare cases and the complication rate remained unaltered. PMID- 16042722 TI - Do all patients with high-grade prostatic intraepithelial neoplasia on initial prostatic biopsy eventually progress to clinical prostate cancer? AB - OBJECTIVE: To assess the clinical outcome of patients with a diagnosis of high grade prostatic intraepithelial neoplasia (PIN) on initial prostatic biopsy, with a minimum of 5 years of follow-up, as such patients are at greater risk of having prostate cancer on subsequent biopsy. PATIENTS AND METHODS: Between November 1992 and October 1998, 21 patients were identified as having PIN on their initial transrectal ultrasonography-guided prostate biopsy. None of these patients had a focus of cancer on the initial biopsy. Their medical data were reviewed retrospectively to determine the natural history of PIN in these patients. Patients who were not identified as having cancer were followed every 6-12 months with prostate-specific antigen (PSA) testing and digital rectal examinations (DRE). RESULTS: A mean (range) of 7 (2-8) cores were taken at initial biopsy; the mean age of the patients was 63 (53-77) years and mean PSA level 9.1 (4.9-17.6) ng/mL. Six patients had an abnormal DRE at presentation. A mean of 8 (7-10) cores were obtained on the second biopsy; six patients were diagnosed with cancer, with a mean Gleason score of 6 (5-7), while three were diagnosed with persistent PIN. These three patients had a third prostate biopsy which showed cancer of Gleason score 6 in one and benign prostatic hyperplasia in two. After a mean follow-up of 72.2 (60-84) months, none of the remaining 12 patients was diagnosed with clinically significant cancer. Five of these patients went on to a third prostate biopsy, with no evidence of cancer. One patient died from unrelated causes during this period. CONCLUSION: This study affirms our current practice of following patients with PIN conservatively if a second or third subsequent prostate biopsy is negative. Whether PIN is a premalignant lesion or merely a lesion associated with cancer needs to be addressed in multicentre studies with a follow-up of > 10 years. PMID- 16042723 TI - Increasing the number of biopsy cores improves the concordance of biopsy Gleason score to prostatectomy Gleason score. AB - OBJECTIVE: To evaluate taking more biopsy cores for predicting the radical prostatectomy (RP) Gleason score compared with the biopsy Gleason score, as although random sextant biopsies are the standard for a tissue diagnosis of prostate cancer, and taking more biopsies increases the detection rate, it is uncertain whether taking more cores improves the prediction of the RP Gleason score. PATIENTS AND METHODS: We analysed retrospectively 404 patients from three centres (Seattle 162, Washington 107 and Chicago 135) who had RP for prostate cancer. Six, eight or 10 biopsies were taken based on the physician's preference and the patient's characteristics. RESULTS: Before RP, 158 (39%) patients had six, 65 (16%) had eight and 181 (45%) had 10 biopsy cores taken. The accuracy of the Gleason sum of the three groups was 65/158 (41%), 26/65 (40%) and 104/181 (57.5%), respectively (P < 0.004, 10-core vs six-core). However, when comparing the Gleason score separately (i.e. 4 + 3 is not equal to 3 + 4), the accuracy of the three groups was 48/158 (30%), 20/65 (31%), and 95/181 (52.5%), respectively (P < 0.001, 10-core vs six core). CONCLUSIONS: Taking more biopsy cores improves the accuracy of the biopsy Gleason score in predicting the final Gleason score at RP; the predictive accuracy of the final Gleason score may be increased from 41% to 58% by increasing the number of biopsies from six to 10. PMID- 16042724 TI - Serum thyroid-stimulating hormone is elevated in men with Gleason 8 prostate cancer. AB - OBJECTIVE: To measure the levels of serum thyroid- stimulating hormone (TSH) in men with prostate cancer, as those with a Gleason score of > or = 8 are at high risk of skeletal metastases (and should be considered for bone scintigraphy at diagnosis), and because the structural integrity of the skeleton depends on constant remodelling controlled by many local and systemic factors, including TSH, an important regulator of this process. PATIENTS AND METHODS: We evaluated 51 men referred for treatment of localized prostate cancer and 10 with biopsy confirmed benign prostatic hypertrophy. Serum TSH was determined with a chemoluminescent immunoassay and a commercially available instrument (Immulite, Diagnostic Products Corporation, Los Angeles). RESULTS: There was significant variation in TSH levels with Gleason score (P = 0.004); men with Gleason 8 tumours had the highest serum TSH levels. Because serum TSH levels increase with age, we used a multivariate analysis of variance with both age and Gleason score as covariates. The effect of Gleason score on TSH level was significant (P = 0.036) and independent of the effect of age (P = 0.392). CONCLUSION: We propose that the high serum TSH levels in men with Gleason 8 prostate cancer is a result of the elaboration of TSH by cancer cells. Bone mineral density in the face of normal levels of thyroid hormone depends on an intact response to TSH, which ordinarily suppresses both osteoblast and osteoclast differentiation, thereby exerting control over bone remodelling. However, with abnormally high TSH levels this process may become deranged, promoting the development of bone metastases. If TSH production by prostate cancer cells could be suppressed, the incidence of bone metastases might be reduced. PMID- 16042725 TI - Inguinal hernia repair with polypropylene mesh during radical retropubic prostatectomy: an easy and practical approach. AB - OBJECTIVE: To report the results of the simultaneous inguinal hernia repair during radical retropubic prostatectomy (RRP) with the preperitoneal tension-free Stoppa technique, using a polypropylene mesh. PATIENTS AND METHODS: During 855 consecutive RRPs, 40 (5%) patients (median age 66.9 years, range 52-81) with 49 inguinal hernias had a simultaneous inguinal hernioplasty. The RRP was performed according to the Walsh modified technique. After the prostate and seminal vesicles were removed and the urethrovesical anastomosis completed, a polypropylene mesh of maximum size 15 x 7.5 cm and a small slit on its medial side was then created and placed in the preperitoneal space, embracing the spermatic cord and covering the myopectinal orifice. Preoperative risk factors, e.g. constipation, pulmonary disease or urinary obstructive symptoms, were collected retrospectively from the files. Complications after surgery, including wound infection, pelvic collections, urinary fistula and recurrence of the hernia, were assessed. RESULTS: Preoperative risk factors for hernia development were identified in 23 (58%) patients; three had recurrent hernias. With a median 23.1 months of follow-up period two (4%) hernias recurred. There were no complications after surgery. CONCLUSION: Preperitoneal hernia repair with polypropylene mesh is safe, effective and practical. The procedure simultaneous with RRP gave a 96% success rate and with no significant increase in operating time or additional complications. PMID- 16042726 TI - An office-based immunodiagnostic assay for detecting urinary nuclear matrix protein 52 in patients with bladder cancer. AB - OBJECTIVE: To report the rapid (5 min) and simple detection of a nuclear matrix protein (NMP) in the urine of patients with bladder cancer, using a newly developed office-based dot-enzyme-linked immunosorbent assay (ELISA). PATIENTS AND METHODS: Western blot and specific immunoglobulin-G antibody were used to identify the urinary NMP marker. Urine samples from 149 patients with bladder cancer and 72 controls were evaluated using the developed dot-ELISA. The initial responses of 43 patients treated by irradiation were followed using the assay. RESULTS: The NMP marker was identified in the urine of patients with bladder cancer at 52 kDa (NMP-52) by Western blot. The dot-ELISA detected the urinary NMP 52 marker in 92% of patients with squamous cell carcinoma, 98% with transitional cell carcinoma, and all six of those with adenocarcinoma of the bladder, with a specificity of 94%. The positive and negative predictive values (97% and 94%, respectively) and efficiency (96%) of the dot-ELISA were high. In addition, the NMP-52 tumour marker was not detected in the urine of patients who showed a response after radiotherapy. CONCLUSION: Detecting the urinary NMP-52 marker using dot-ELISA would be helpful in the rapid diagnosis and follow-up of patients with bladder cancer. PMID- 16042727 TI - Detrusor myectomy: long-term results with a minimum follow-up of 2 years. AB - OBJECTIVES: To assess the long-term results of detrusor myectomy, which has obvious theoretical advantages over enterocystoplasty for refractory detrusor overactivity (DO), and for which the early results have been encouraging. PATIENTS AND METHODS: The medical records were reviewed of 30 consecutive patients (median age 33 years, range 10-62) who had a detrusor myectomy between November 1992 and April 2002 in our unit. Twenty-four patients (80%) had idiopathic DO (six males and 18 females) and six (20%) had neurogenic DO (four males and two females). The median (range) follow-up was 79 (28-142) months. All patients were confirmed to have DO on urodynamics before surgery and 26 (87%) had urodynamics afterward. RESULTS: Nineteen (79%) of those with idiopathic DO and two with neurogenic DO showed a continued overall improvement. The cystometric capacity improved in 80% of patients after surgery, whilst the detrusor pressure at maximum flow and the bladder contractility index decreased in 60% and 78% of the patients, respectively. Ten patients (45%) had to start clean intermittent self-catheterization after surgery. CONCLUSIONS: Detrusor myectomy is successful in approximately 80% of patients with idiopathic DO, although detrusor contractility is affected in most and almost half of the patients required clean intermittent self-catheterization afterward. This procedure should be offered as an alternative to enterocystoplasty as it is less morbid and does not preclude subsequent surgery if required. However, further evaluation of this technique is required in neuropathic patients. PMID- 16042728 TI - Are conventional pressure-flow measurements dependent upon filled volume? AB - OBJECTIVE: To determine, in a prospective study, whether detrusor pressure (p(det.Qmax)) and maximum urinary flow rate (Q(max)) measurements obtained after filling to maximum cystometric capacity (MCC) differ from those obtained with filling restricted to average voided volume (V(void)), as standard protocols for pressure flow studies (PFS) mandate bladder filling until the subject has a strong desire to void, which aids standardization but further divorces the test from real-life experience. PATIENTS AND METHODS: After calculating the appropriate sample size, 84 patients attending for PFS with an adequately completed 3-day frequency-volume chart were recruited. Each underwent two consecutive PFS with filling to MCC and average V(void) in a random order, and measurements of p(det.Qmax) and Q(max) were compared. For men, the agreement for a diagnosis of obstruction between the tests was also assessed. RESULTS: Complete data were obtained from 76 (90%) of the patients, with a mean (range) age of 64 (20-94) years. The mean (sd) difference between MCC and average V(void) was 134 (113) mL (P < 0.01). There were no significant differences between estimates of Q(max), at - 0.1 (3) mL/s (P = 0.75), and of p(det.Qmax), at - 1 (13) cmH(2)O (P = 0.91), obtained within each patient. For men there was 91% agreement (32 of 35) in the classification of obstruction. CONCLUSIONS: Restriction of filling to the average V(void) during PFS allows a closer approximation to normal voiding and results in no clinically relevant change to the value of standard pressure-flow measurements or alters individual classification of obstruction. PMID- 16042729 TI - Validation of a patient-administered questionnaire to measure the severity and bothersomeness of lower urinary tract symptoms in uncomplicated urinary tract infection (UTI): the UTI Symptom Assessment questionnaire. AB - OBJECTIVE: To develop and validate a self-administered questionnaire to assess the 'severity' and 'bothersomeness' of the most frequently reported signs and symptoms of uncomplicated urinary tract infection (uUTI). SUBJECTS AND METHODS: The UTI Symptoms Assessment questionnaire (UTISA) is a 14-item instrument asking about the severity and bothersomeness of seven key uUTI symptoms. It was developed after comprehensive literature and data review and administration in draft form to a sample of 30 women with uUTI. The final questionnaire was completed by 276 women with uUTI who participated in a noncomparative clinical trial of ciprofloxacin. The women completed the questionnaire in electronic format at baseline (before the first dose of ciprofloxacin once-daily), at 3-h and 8-h intervals until all UTI symptoms were resolved, and at the test-of-cure visit. Baseline scores on the King's Health Questionnaire (KHQ) were used to assess convergent and divergent validity; responses to the Global Rating of Change (GRC) were used to assess both responsiveness and the 'minimally important difference'. Discriminant validity and responsiveness were assessed by comparing UTISA scores with a clinical evaluation of UTI symptoms performed by the investigator at baseline and at the test-of-cure visit. RESULTS: The UTISA was found to comprise three four-item domains named 'urination regularity', 'problems with urination', and 'pain associated with UTI'. Two questions asking about haematuria loaded on a fourth factor. The three domains were homogeneous (with high inter-item correlations) and internally consistent. Convergent validity was shown by high correlations between similar UTISA and KHQ domains (all r(s) > 0.40), and divergent validity by small correlations between unlike domains (all r(s) < 0.15). In general, the UTISA domains showed excellent discriminant validity, with scores on selected domains discriminating between women with different clinical evaluations. The responsiveness of the UTISA was also excellent, with high correlations between changes in domain scores and the clinical evaluation and GRC items. Symptom improvement was highest in the first 3 h, leading to greater responsiveness and minimally important difference during this period. However, the UTISA could detect even small subsequent changes. CONCLUSION: The three-domain UTISA has excellent psychometric properties and it is likely to prove an excellent tool for assessing uUTI outcome from a patient's perspective, both in research and clinical settings. PMID- 16042730 TI - A novel midstream urine-collection device reduces contamination rates in urine cultures amongst women. AB - OBJECTIVE: To evaluate a novel urine-collection device (UCD) that automatically collects a midstream urine (MSU) sample, and compare contamination rates to those of the conventional MSU sampling method, as the contamination of urine samples for microbiological analysis in women leads to diagnostic ambiguity and unnecessary costs, and may result in part from an incorrect collection procedure. PATIENTS AND METHODS: In all, 2823 women from four centres, most from antenatal clinics, were randomized to two urine-collection methods: conventional MSU collection and collection with a novel MSU UCD (the Whiz, JBOL Ltd, Oxford, UK). Semi-quantitative growth and user acceptability were compared between the collection methods. RESULTS: MSU samples collected with the UCD had significantly fewer mixed growth samples (9% vs 14%, P = 0.001; 36% relative reduction), significantly fewer heavy mixed growth samples (1.2% vs 3.0%, P = 0.004; 60% relative reduction) and required significantly fewer re-tests (11% vs 16%, P = 0.002; 31% relative reduction). There were more samples with clinically insignificant growth than the conventional MSU group (86% vs 82%, P = 0.005). Those using the UCD preferred it to the conventional method (67.5%) and experienced significantly less spillage during sample collection (27% vs 46%, P = 0.001; relative reduction 41%). CONCLUSION: The UCD reduced contamination rates in urine samples and improved the predictive value of the urine culture in a manner acceptable to patients and staff. PMID- 16042731 TI - How do urinary diaries of women with an overactive bladder differ from those of asymptomatic controls? AB - OBJECTIVE: To quantify clinically important differences in common diary variables between asymptomatic controls and women with symptoms of overactive bladder (OAB), controlling for the effects of age and race. PATIENTS, SUBJECTS AND METHODS: The 24-h urinary diaries of 49 women with symptoms of OAB were compared to those of age- and race-matched asymptomatic controls. Control subjects did not have a physical examination. RESULTS: The 49 patients with OAB symptoms had a median (range) age of 51 (20-85) years, a body mass index of 25 (17-46) kg/m(2) and a parity of 2 (0-5). The median number of voids was significantly greater in women with OAB than asymptomatic controls (P < 0.001). The median value for mean voided volume was significantly lower in women with OAB than asymptomatic controls (P = 0.014). There was no difference in the maximum voided volume, total voided volume, daytime or night-time diuresis rates, voids per litre intake, or total fluid intake. CONCLUSIONS: This preliminary study suggests that a median reduction of three voids/24 h and an increase of 70 mL in the mean voided volume might be clinically important goals in therapeutic trials for treating OAB symptoms. This remains to be confirmed by further studies linking improvements in quality of life and the overall impression of bladder health with these quantitative diary variable changes. PMID- 16042732 TI - A comparison of the effect of 1.5% glycine and 5% glucose irrigants on plasma serum physiology and the incidence of transurethral resection syndrome during prostate resection. AB - OBJECTIVE: To examine changes in the pathophysiology and frequency of the transurethral resection (TUR) syndrome with two irrigation fluids, as variable amounts of irrigation fluid are absorbed during TUR of the prostate (TURP), and although polar solutes are required to prevent an effect on diathermy, the solutes may have effects when absorbed. PATIENTS AND METHODS: Between December 2001 and March 2003, 250 patients were included in a prospective randomized trial comparing glycine 1.5% with 5% glucose irrigation fluids. We measured blood loss, fluid absorption, temperature change, biochemistry including a glycine assay, and peri-operative symptoms. Blood samples were taken immediately before and immediately, 5 and 24 h after TURP. Irrigating fluid absorption during TURP was measured with 1% ethanol as a marker and breath ethanol measurements. Operative details were recorded, including the type of anaesthesia (with or with no sedation), resection time and weight of resected tissue. Peri-operative symptoms were documented prospectively. TUR syndrome was defined as a serum sodium level of < or = 125 mmol/L with two or more associated symptoms or signs of TUR syndrome. RESULTS: Five (2%) patients had TUR syndrome; all five were irrigated with glycine, although this difference was not statistically significant (P = 0.06). Of the five men, three had hypotension, four were tired, one was nauseous, two had parasthesia, two had 'uneasiness', one had blurred vision and two were confused; none had chest pain. There was a large variation between the groups in the level of glycine assayed immediately after TURP; a high glycine level was associated with the TUR syndrome (P = 0.01). There was no difference between the groups in levels of sodium, potassium, urea, creatinine, osmolality, calcium, haematocrit, albumin serum levels or peri-operative blood loss (defined as a change from before to after TURP in haemoglobin level, accounting for transfusions). CONCLUSIONS: An increase in serum glycine was associated with TUR syndrome; there were large variations in the amounts of glycine absorbed, reaching levels many times the upper limit of normal. In other studies, glycine was reportedly toxic, and that the levels recorded were many times the upper limit of normal may have both immediate and long-term effects. PMID- 16042733 TI - The management of penile fracture based on clinical and magnetic resonance imaging findings. AB - OBJECTIVE: To present our experience with repairing penile fracture, based on clinical and magnetic resonance imaging (MRI) findings. PATIENTS AND METHODS: Between December 2002 and October 2004, 14 men (19-64 years old) presented to our centre with a penile fracture. Two patients had urethral bleeding. MRI was used before surgery in all patients, and the repair comprised a localized longitudinal penile incision in 13 men. This incision was designed according to the tunical tear site and size already depicted by MRI. One case was managed conservatively, as MRI confirmed an intercavernosal haematoma with no tunical tear. The follow-up was 4-21 months. RESULTS: The tear involved one corpus cavernosum in 11 patients; two were associated with urethral injury. The course after repair was uneventful in all men; the follow-up showed no erectile dysfunction in any. The patients reported neither pain nor penile curvature during erection. CONCLUSION: MRI is a simple and informative investigation for evaluating and documenting a penile fracture, and it improves the management plan. PMID- 16042734 TI - Outcome from percutaneous nephrolithotomy in patients with spinal cord injury, using a single-stage dilator for access. AB - OBJECTIVE: To present our experience of percutaneous nephrolithotomy (PCNL) for treating urolithiasis in patients with spinal cord injury (SCI) using a single stage dilator for percutaneous access. PATIENTS AND METHODS: A prospective database of patients with SCI having PCNL using the single-stage dilator was assessed, analysing patient data, stone-free rates, morbidity and the follow-up outcome. RESULTS: In all, 26 patients had 54 PCNLs on 32 kidneys; 20 had unilateral and six bilateral stone disease; there were many staghorn calculi (24/54). Major complications occurred in three of 54 PCNLs (6%). The complete stone-clearance rate was 87% for PCNL alone, rising to 29 of 32 kidneys (91%) or 24 of 26 patients (92%) with adjuvant procedures. A further three kidneys required no further treatment and were monitored, having residual fragments of < or = 2 mm. CONCLUSIONS: PCNL has a high success rate and acceptable complication rate compared to extracorporeal shock-wave lithotripsy, and remains a valid first line treatment option for kidney stones in patients with SCI. PMID- 16042735 TI - Multimodal management of urolithiasis in renal transplantation. AB - OBJECTIVE: To report the largest single series of renal transplant patients (adults and children) with urolithiasis, assess the risk factors associated with urolithiasis in renal transplant recipients, and report the outcome of the multimodal management by endourological and open procedures. PATIENTS AND METHODS: The records of all patients undergoing renal transplantation between 1977 and 2003 were reviewed. In all, 2085 patients had a renal transplant at our centre and 21 (17 adults and four children) developed urinary tract calculi. Their mode of presentation, investigations, treatments, complications and outcomes were recorded. Investigations included one or more of the following; ultrasonography (US), plain abdominal X-ray, intravenous urography, nephrostogram and computed tomography. Management of these calculi involved extracorporeal shock wave lithotripsy (ESWL), flexible ureteroscopy and in situ lithotripsy, percutaneous nephrolithotomy (PCNL), open pyelolithotomy and open cystolitholapaxy. RESULTS: Thirteen patients had renal calculi, seven had ureteric calculi and one had bladder calculi. The incidence of urolithiasis was 21/2085 (1.01%) in the series. Urolithiasis was incidentally discovered on routine US in six patients, six presented with oliguria or anuria, including one with acute renal failure, four with a painful graft, three with haematuria, one with sepsis secondary to obstruction and infection and in one, urolithiasis was found after failure to remove a stent. Ten patients (63%) had an identifiable metabolic cause for urolithiasis, two by obstruction, two stent-related, one secondary to infection and in six no cause was identifiable. Thirteen required more than one treatment method; 13 (69%) were treated by ESWL, eight of whom required multiple sessions; eight required ureteric stent insertion before a second procedure and four required a nephrostomy tube to relieve obstruction. Two patients had flexible ureteroscopy and stone extraction, three had a PCNL and one had open cystolithotomy. PCNL failed in one patient who subsequently had successful open pyelolithotomy. All patients were rendered stone-free when different treatments were combined. CONCLUSIONS: The incidence of urolithiasis in renal transplant patients is low. There is a high incidence of metabolic causes and therefore renal transplant patients with urolithiasis should undergo comprehensive metabolic screening. Management of these patients requires a multidisciplinary approach by renal physicians, transplant surgeons and urologists. PMID- 16042736 TI - Asymptomatic bacteriuria in men with orthotopic ileal neobladders: possible relationship to nocturnal enuresis. AB - OBJECTIVE: To assess prospectively the incidence with time of asymptomatic bacteriuria in patients with orthotopic ileal neobladders, and the possible effect on neobladder function. PATIENTS AND METHODS: In all, 47 patients (mean age 52.7 years, sd 8.7, range 31-68) with uncomplicated orthotopic ileal neobladders were prospectively evaluated. With no antibiotic manipulation, consecutive urine cultures were assessed monthly. Continence was assessed by direct information from the patients at each follow-up visit. RESULTS: Overall, 797 samples were cultured from the 47 patients (mean 17.6, sd 7.1). There was a steady decrease in the incidence of positive cultures, from 74.5%, to 35.6% and 6.7% at 1, 6 and 18 months, respectively. While there was persistently sterile urine in only eight patients (17%), 32 had occasional and seven had persistent bacteriuria. Escherichia coli was the commonest organism (76.6%) followed by Klebsiella pneumonia (15.7%); 54% of E. coli and 38% of K. pneumonia infections were sensitive to nitrofurantoin. Diurnal continence was achieved in 98% of the patients at 6 months after surgery. There was a gradual decrease in the frequency of nocturnal enuresis (NE) with time, from 87%, to 42%, 28% and 27% at 1, 6, 12 and 18 months, respectively. There was a significant correlation between the presence of bacteriuria and NE during the first 6 months, but it was not sustained after that. The age of the patients was also related significantly to the incidence of NE; at 6 months, only one of 18 men aged < or = 50 years had NE, while 19 of 29 aged > 50 years had (P < 0.001). At 1 year all patients aged < or = 50 years were nocturnally continent, while half of those aged > 50 years had NE (P = 0.001). CONCLUSIONS: Ileal neobladders are associated with a high incidence of asymptomatic bacteriuria during the first year after surgery. There was spontaneous clearance of bacteriuria with time, with no antimicrobial manipulation. Soon after surgery there was a significant association between bacteriuria and NE. The effect of antimicrobials on patients with NE should be evaluated. PMID- 16042737 TI - Vesicostomy revisited: the best treatment for the hostile bladder in myelodysplastic children? AB - OBJECTIVE: To evaluate the effects of vesicostomy on the urinary tract of myelodysplastic children in whom conservative bladder management with clean intermittent catheterization (CIC) has failed to preserve upper and lower urinary tract function. PATIENTS AND METHODS: Sixteen children with myelodysplasia underwent vesicostomy. Indications included worsening hydronephrosis, vesico ureteric reflux (VUR), recurrent urinary tract infections (UTIs), and increasing renal insufficiency despite CIC and/or difficulty with CIC. The mean (range) age at vesicostomy was 36.5 (9-82) months and the follow-up 7.4 (2-16) years. RESULTS: Hydronephrosis resolved or improved in 12 of 14 children, the incidence of UTI decreased to one or fewer per year in 10, VUR resolved or improved in nine, and renal function improved or stabilized in six of seven patients. One patient initially presented with renal insufficiency and subsequently required dialysis despite vesicostomy. Complications occurred in three of 15 children, and included stomal stenosis and bladder calculi. The vesicostomy was closed in six patients after a mean of 4.4 (1.5-9) years. Four of these patients required concomitant bladder augmentation. CONCLUSIONS: Vesicostomy in myelodysplastic children is effective in preventing and/or resolving the deleterious consequences of a 'hostile' bladder. The procedure is uncomplicated, well tolerated, reversible and should be considered in managing children in whom conservative management by CIC has failed. PMID- 16042738 TI - Inguinal hernia in female infants: a cue to check the sex chromosomes? AB - OBJECTIVES: To review the clinical presentation of complete androgen insensitivity syndrome (CAIS) and assess the current practice of considering the diagnosis of CAIS in female infants presenting with inguinal hernia. PATIENTS AND METHODS: AIS arises from target tissue resistance to the actions of androgens due to molecular abnormalities in the androgen receptor. Patients with CAIS are born with normal external female genitalia, and although inguinal hernias are uncommon in female infants, they are a well-known presentation of CAIS. Such patients were identified from the Cambridge Intersex Database and details of presentation, presence and laterality of inguinal hernia and contents, and family history of CAIS, were recorded. A questionnaire detailing different indications for considering CAIS in female infants with a hernia was distributed to members of the British Association of Paediatric Surgeons and the British Society for Paediatric Endocrinology and Diabetes. RESULTS: More than half of patients with CAIS presented with inguinal hernia, of which half were bilateral and a third contained gonads. Completed questionnaires were returned by 87 surgeons and 64 endocrinologists, and most of the surgeons and endocrinologists would consider CAIS in all female infants with a hernia. Bilateral hernias, hernias containing gonads and a family history of CAIS would prompt clinicians to consider the diagnosis. CONCLUSION: Most clinicians agreed that CAIS should be considered in all female infants with inguinal hernia, as this is the commonest mode of presentation in childhood. Macroscopic inspection of the internal genital structures coupled, perhaps, with gonadal biopsy is recommended. Fluorescence in situ hybridization offers a rapid and reliable method to check the sex chromosomes. Liaison between the paediatric surgeon and endocrinologist is essential in management of infants with CAIS. PMID- 16042739 TI - Nocturnal enuresis at 7.5 years old: prevalence and analysis of clinical signs. AB - OBJECTIVE: To determine the prevalence of nocturnal enuresis (NE) in a large cohort of children at 7.5 years old, and to examine the frequency of variables such as gender, severity, associated elimination problems, and clinical signs within the identified group. SUBJECTS AND METHODS: Of an original cohort of 13 971 infants alive at 12 months, 11 251 who were still active in the Avon Longitudinal Study of Parents and Children (ALSPAC) survey, were followed at 91 months. The mother or main carer was given a questionnaire which asked, amongst other items, about the presence and frequency of bedwetting, other elimination problems, and signs related to the wetting behaviour; 8269 (73.5%) questionnaires were returned and 8151 contained information on the frequency of bedwetting. RESULTS: In all, 1260 children (15.5%) at 7.5 years wet the bed, but most wet once or less a week, and only 215 (2.6%) met the Diagnostic and Statistical Manual of Mental Disorders (fourth edition) criteria of NE (wetting at least twice a week). A higher prevalence was reported in boys than girls and 266 children (3.3%) had both daytime wetting and bedwetting, with 189 (2.3%) having both daytime soiling and bedwetting. Daytime urgency increased with severity of bedwetting and occurred in 28.9% of children with NE. CONCLUSION: At 7.5 years old the incidence of bedwetting is high, but only 2.6% of this large population based sample wet at a frequency meeting the definition of NE. Although a small percentage of children had both daytime wetting and bedwetting, the evidence suggests that these are discrete problems. Amongst children with NE, indicators of bladder overactivity were present, supporting the view of heterogeneity and the importance of individual assessment in deciding on appropriate treatment. PMID- 16042740 TI - Efficacy of tolterodine as a first-line treatment for non-neurogenic voiding dysfunction in children. AB - OBJECTIVE: To assess the effect of antimuscarinic treatment with tolterodine combined with behavioural modification as a first-line treatment, before invasive investigation, in children with non-neurogenic voiding dysfunction but no obvious anatomical or neurogenic cause. PATIENTS AND METHODS: The study comprised 44 children presenting with voiding dysfunction (30 girls and 14 boys, mean age 7 years, range 5-14); all had a noninvasive evaluation consisting of a history, urine analysis, renal and bladder ultrasonography and physical examination, with specific emphasis on the voiding pattern. Anticholinergic treatment with tolterodine (1 mg twice daily) was started in all patients; they were also informed about conservative management, including timed voiding, double voiding and relaxation of the pelvic floor during voiding. At the start and after 3 months, the dysfunctional voiding symptom score (DVSS) was completed twice by all patients. RESULTS: For all patients the mean (sd) DVSS was 14.0 (2.67) and 6.68 (3.67) before and after treatment, respectively; the difference was statistically significant (P < 0.001). The mean scores for girls and boys, respectively, were 13.8 (2.79) and 14.5 (2.44) before and 6.43 (3.79) and 7.50 (3.34) after treatment. CONCLUSION: Tolterodine combined with behavioural modification for dysfunctional voiding in children with no neurological or anatomical abnormality can be recommended as a first-line treatment before invasive evaluation. Additionally, the DVSS appears to provide accurate and objective data for monitoring the effect of treatment in such children. PMID- 16042741 TI - The src-family kinase inhibitor PP2 suppresses the in vitro invasive phenotype of bladder carcinoma cells via modulation of Akt. AB - OBJECTIVE: To evaluate PP2 as a modulator of the cadherin/catenin complex in late stage bladder carcinoma cells, and to assess its potential invasion-suppressor activity in this model. MATERIALS AND METHODS: A panel of five human bladder carcinoma cells, characterizing late-stage disease, was used to determine the concentration for 50% inhibition of PP2 in cell-proliferation assays. Modulation of cadherin/catenin expression by PP2 was determined in Western blot analysis, with an assessment of the activation status of mitogen-activated protein kinase and Akt signalling pathways. Altered invasive capacity linked to these variables was determined in standard in vitro invasion assays. RESULTS: PP2 elicited concentration-dependent growth inhibition in all bladder cell lines within the panel, with growth suppression recorded at 10-35 micromol/L PP2. Distinct morphological changes were recorded in cell lines exposed to PP2, accompanied by up-regulation of plakoglobin expression in a subset of lines. Exposure of cells to PP2 resulted in inactivation of Akt in all cells and a concomitant reduction in in vitro invasive capacity. CONCLUSIONS: These results show that PP2 inhibits bladder carcinoma cell growth and can modulate plakoglobin expression in a subset of cell lines. In addition, PP2 can suppress the in vitro invasive capacity of bladder carcinoma cells by modulating the activation status of Akt. PMID- 16042742 TI - Sildenafil inhibits the formation of superoxide and the expression of gp47 NAD[P]H oxidase induced by the thromboxane A2 mimetic, U46619, in corpus cavernosal smooth muscle cells. AB - OBJECTIVE: To assess the effect of sildenafil on superoxide formation and p47(phox) (the active subunit of NADPH oxidase) expression in cultured corpus cavernosal smooth muscle cells (CVSMCs). MATERIALS AND METHODS: CVSMCs derived from rabbit penis were incubated with U46619 (thromboxane A2 analogue) with or without sildenafil for 1 or 16 h at 37 degrees C. Superoxide dismutase inhibitable superoxide formation was assessed using the reduction of ferricytochrome c measured spectrophotometrically, and gp47(phox) assessed using Western blot analysis. The role of NAD[P]H oxidase and cGMP was further studied by using specific inhibitors of each. RESULTS: Superoxide formation was significantly greater in cells incubated with U46619 after 1 and 16 h incubation than in controls, an effect blocked by NADP(H) oxidase inhibitors. These effects of U46619 were inhibited by sildenafil (1 and 10 nmol/L), which in turn were negated by the guanylyl cyclase inhibitor, ODQ; 10 nmol/L sildenafil inhibited p47phox expression induced by U46619. CONCLUSIONS: Sildenafil is a potent inhibitor of superoxide formation in CVSMCs. This effect is mediated through the inhibition of PDE-5 which in turn augments the inhibitory action of the NO-cGMP axis on NAD[P]H oxidase expression and activity. This mechanism constitutes a new pharmacological action of sildenafil, consolidates the potential role of superoxide in ED, and indicates that thromboxane A(2) may be an important mediator of intrapenile oxidative stress. PMID- 16042743 TI - Loss of ryanodine receptor calcium-release channel expression associated with overactive urinary bladder smooth muscle contractions in a detrusor instability model. AB - OBJECTIVE: To investigate the changes in spontaneous bladder smooth muscle contractions that occur during detrusor instability (DI), and to test the possibility that altered function or expression of ryanodine receptors (RyRs) could account for the increased bladder contractions. MATERIALS AND METHODS: After 8 weeks of partial bladder outlet obstruction, DI was confirmed in female experimental rats by filling cystometry. Muscle strips were dissected from freshly isolated bladders, and isometric tension recorded in strips from DI and normal bladders. The contractions were recorded during electrical stimulation or exposure to various agents. Western blot analysis was used to determine RyR expression in DI and normal bladder muscle. RESULTS: In DI bladder muscle, spontaneous contractile activity persisted in the presence of blockers for known neurotransmitter receptors in the bladder wall. The RyR blocker ryanodine significantly increased the spontaneous contractile frequency in normal bladder strips, but failed to affect spontaneous contractions in DI muscle. Caffeine inhibited spontaneous contractile activity in both the DI and normal strips. After administering the l-type Ca(2+) channel antagonist nimodipine, the myogenic contractile activity was abolished in normal strips; in contrast, in DI strips, the amplitude of contractions was reduced but the frequency of contractions was unchanged. Western blot analysis showed that RyR expression was lower in DI muscle than in normal bladder muscle. CONCLUSION: These results provide the first characterization of a loss of regulation of spontaneous contractile activity by RyRs in DI muscle associated with a significant decrease in RyR expression. RyRs in normal detrusor muscle act as negative-feedback regulators of spontaneous contractile activity, presumably by releasing Ca(2+) that activates Ca(2+) dependent K(+) channels to decrease contractility. This mechanism might be weakened in DI muscle, resulting in spontaneous contractile overactivity. PMID- 16042744 TI - Unseen forces at AUA 2005? PMID- 16042745 TI - A simple modification to the Albarran deflector enhances endoscopic control. PMID- 16042746 TI - Cystoscopic removal of a JJ stent using a suture 'lasso'. PMID- 16042748 TI - Analysis of HER2 expression in primary urinary bladder carcinoma and corresponding metastases. PMID- 16042749 TI - The role of urinary urgency and its measurement in the overactive bladder symptom syndrome: current concepts and future prospects. PMID- 16042750 TI - Laparoscopy for impalpable testes. PMID- 16042752 TI - C-reactive protein is significantly associated with prostate-specific antigen and metastatic disease in prostate cancer. PMID- 16042753 TI - The incidence and treatment of lymphoceles after radical retropubic prostatectomy. PMID- 16042754 TI - Simplified orthotopic ileocaecal pouch (Mainz pouch) for bladder substitution. PMID- 16042756 TI - A novel alternative splicing form of excitatory amino acid transporter 1 is a negative regulator of glutamate uptake. AB - Abstract EAAT1 is a major glutamate transporter in the CNS and is required for normal neurotransmission and neuroprotection from excitotoxicity. In the present study, we have identified a novel form of the human EAAT1, named here as EAAT1ex9skip, which lacks the entire exon 9. Quantitative PCR analysis indicates that this variant is expressed throughout the CNS, both in grey matter and axonal tracts, at levels ranging between 10% and 20% of the full-length EAAT1 form. When expressed in HEK293 cells, EAAT1ex9skip mRNA is translated into a truncated protein localized in the endoplasmic reticulum. EAAT1ex9skip has no functional glutamate uptake activity but instead, exerts a dominant negative effect over full-length EAAT1 function. In turn, co-expression of full-length EAAT1 and EAAT1ex9skip variants reduces the insertion of the former into the plasma membrane. Together, these results indicate that the EAAT1ex9skip splice variant is a negative regulator of full-length EAAT1 function in the human brain. PMID- 16042757 TI - Glioma cells under hypoxic conditions block the brain microvascular endothelial cell death induced by serum starvation. AB - Angiogenesis is one of essential components for the growth of neoplasms, including malignant gliomas. However, tumor vascularization is often poorly organized and marginally functional due to tumor structural abnormalities, inducing regional or temporal hypoxic conditions and nutritional shortages in tumor tissues. We investigated how during angiogenesis migrating endothelial cells survive in these hypoxic and reduced nutritional conditions. Human brain microvascular endothelial cells (HBMECs) underwent apoptosis and necrosis after serum withdrawal. This endothelial cell death was blocked by recombinant VEGF protein or the culture medium of U251 glioma cells exposed to hypoxia (H-CM). Hypoxic treatment increased vascular endothelial growth factor (VEGF) and tumor necrosis factor alpha (TNF-alpha) expression in U251 glioma cells. H-CM activated nuclear factor-kappaB (NFkappaB) protein and increased the gene expression of antiapoptotic factors including Bcl-2, Bcl-X(L), survivin and X-chromosome-linked inhibitor of apoptosis protein (XIAP) in endothelial cells. The survival activity of H-CM for endothelial cells was abolished by two kinds of VEGF inhibitors {Cyclopeptidic VEGF inhibitor and a VEGF receptor tyrosine kinase inhibitor (4 [(4'-chloro-2'-fluoro) phenylamino]-6, 7-dimethoxyquinazoline)} or NFkappaB inhibitors (ALLN and BAY 11-7082). These VEGF inhibitors did not block the activation of NFkappaB induced by H-CM in endothelial cells. On the contrary, TNF alpha antagonist WP9QY enhanced the survival activity of H-CM for endothelial cells and blocked NFkappaB activation induced by H-CM under serum-starved conditions. Taken together, our data suggest that both the secretion of VEGF from glioma cells and activation of NFkappaB in endothelial cells induced by TNF-alpha are necessary for endothelial cell survival as they increase the expression of antiapoptotic genes in endothelial cells under conditions of serum starvation. These pathways may be one of the mechanisms by which angiogenesis is maintained in glioma tissues. PMID- 16042758 TI - Functions and pathophysiological roles of phospholipase D in the brain. AB - Ten years after the isoforms of mammalian phospholipase D (PLD), PLD1 and 2, were cloned, their roles in the brain remain speculative but several lines of evidence now implicate these enzymes in basic cell functions such as vesicular trafficking as well as in brain development. Many mitogenic factors, including neurotransmitters and growth factors, activate PLD in neurons and astrocytes. Activation of PLD downstream of protein kinase C seems to be a required step for astroglial proliferation. The characteristic disruption of the PLD signaling pathway by ethanol probably contributes to the delay of brain growth in fetal alcohol syndrome. The post-natal increase of PLD activities concurs with synapto- and myelinogenesis in the brain and PLD is apparently involved in neurite formation. In the adult and aging brain, PLD activity has antiapoptotic properties suppressing ceramide formation. Increased PLD activities in acute and chronic neurodegeneration as well as in inflammatory processes are evidently due to astrogliosis and may be associated with protective responses of tissue repair and remodeling. ARF-regulated PLD participates in receptor endocytosis as well as in exocytosis of neurotransmitters where PLD seems to favor vesicle fusion by modifications of the shape and charge of lipid membranes. Finally, PLD activities contribute free choline for the synthesis of acetylcholine in the brain. Novel tools such as RNA interference should help to further elucidate the roles of PLD isoforms in brain physiology and pathology. PMID- 16042759 TI - Variation in gene expression patterns in effusions and primary tumors from serous ovarian cancer patients. AB - BACKGROUND: While numerous studies have characterized primary ovarian tumors, little information is available regarding expression patterns of metastatic sites of this cancer. To define sets of genes that distinguish primary and metastatic ovarian tumors, we used cDNA microarrays to characterize global gene expression patterns in 38 effusions (28 peritoneal, 10 pleural) and 8 corresponding primary ovarian tumors, and searched for associations between expression patterns and clinical parameters. RESULTS: We observed multidimensional variation in expression patterns among the cancers. Coordinate variation in expression of genes from two chromosomal regions, 8q and 19q, was seen in subsets of the cancers indicating possible amplifications in these regions. A set of 112 unique genes of known function was differentially expressed between primary tumors and effusions using supervised analysis. Relatively few differences were seen between effusions isolated from the pleural and peritoneal cavities or between effusions from patients diagnosed with stage III and stage IV cancers. A set of 84 unique genes was identified that distinguished high from lower grade ovarian cancers. The results were corroborated using immunocytochemistry, mRNA in situ hybridization, and immunoblotting. CONCLUSION: The extensive variation in expression patterns observed underscores the molecular heterogeneity of ovarian cancer, but suggests a similar molecular profile for ovarian carcinoma cells in serosal cavities. PMID- 16042760 TI - Serum biomarkers in interstitial lung diseases. AB - The use of biomarkers in medicine lies in their ability to detect disease and support diagnostic and therapeutic decisions. New research and novel understanding of the molecular basis of the disease reveals an abundance of exciting new biomarkers who present a promise for use in the everyday clinical practice. The past fifteen years have seen the emergence of numerous clinical applications of several new molecules as biologic markers in the research field relevant to interstitial lung diseases (translational research). The scope of this review is to summarize the current state of knowledge about serum biomarkers in interstitial lung diseases and their potential value as prognostic and diagnostic tools and present some of the future perspectives and challenges. PMID- 16042761 TI - Severely disabling chronic pain in young adults: prevalence from a population based postal survey in North Staffordshire. AB - BACKGROUND: Severely disabling chronic pain in the adult population is strongly associated with a range of negative health consequences for individuals and high health care costs, yet its prevalence in young adults is less clear. METHODS: All adults aged 18-25 years old registered with three general practices in North Staffordshire were invited to complete a postal questionnaire containing questions on pain within the last 6 months, pain location and duration. Severity of chronic pain was assessed by the Chronic Pain Grade. Severely disabling chronic pain was defined as pain within the last six months that had lasted for three months or more and was highly disabling-severely limiting (Grade IV). RESULTS: 858 responses from 2,389 were received (adjusted response = 37.0%). The prevalence of any pain within the previous six months was 66.9% (95%CI: 63.7%, 70.1%). Chronic pain was reported by 14.3% (95%CI: 12.0%, 16.8%) of respondents with severely disabling chronic pain affecting 3.0% (95%CI: 2.0%, 4.4%) of this population. Late responders were very similar to early responders in their prevalence of pain. Cross-checking the practice register against the electoral roll suggested register inaccuracies contributed to non-response. CONCLUSION: Pain is a common phenomenon encountered by young adults, affecting 66.9% of this study population. Previously observed age-related trends in severely disabling chronic pain in older adults extend to younger adults. Although a small minority of younger adults are affected, they are likely to represent a group with particularly high health care needs. High levels of non-response in the present study means that these estimates should be interpreted cautiously although there was no evidence of non-response bias. PMID- 16042762 TI - Validation of the Italian version of the "Mood Disorder Questionnaire" for the screening of bipolar disorders. AB - The study measured the accuracy of the Italian version of the Mood Disorder Questionnaire (MDQ) as a screening instrument for bipolar disorders in a psychiatric setting. METHODS: 154 consecutive subjects attending the Division of Psychiatry of the University of Cagliari (Italy), were screened for bipolar disorders using the Italian translation of the MDQ, and diagnostically interviewed with the Structured Clinical Interview for DSM-IV Axis I Disorders (SCID) by physicians. RESULTS: On the basis of the SCID: 51 (33.1%) received a diagnosis of bipolar or schizoaffective bipolar type disorders, 63 (40.9%) were diagnosed as having at least one psychiatric disorder in Axis I (other than bipolar or schizoaffective bipolar type disorders), whilst 40 (25.9%) were unaffected by any type of psychiatric disorder. MDQ showed a good accuracy for bipolar or schizoaffective bipolar type disorders: the cut-off 4 had sensitivity 0.90 and specificity 0.58; the cut-off 5 had sensitivity 0.84 and specificity 0.70; and the cut-off 6 had sensitivity 0.76 and specificity 0.86. The accuracy for bipolar II disorders was sufficient but not excellent: the cut-off 4 had sensitivity 0.80 and specificity 0.45; the cut-off 5 had sensitivity 0.70 and specificity 0.55; and the cut-off 6 had sensitivity 0.55 and specificity 0.65. CONCLUSION: Our results seem to indicate a good accuracy of MDQ, and confirm the results of recent surveys conducted in the USA. Moreover the instrument needs to be validated in other settings (e.g. in general practice). PMID- 16042763 TI - Level of positive mental health in the European Union: results from the Eurobarometer 2002 survey. AB - BACKGROUND: Few epidemiological studies have focused on the occurrence of positive mental health, and those comparing several countries practically non existent. This study presents comparative findings of positive mental health in 11 EU countries or regions, based on the Eurobarometer 2002 (autumn) survey. METHOD: The sample (n = 10,878) represents the general population, aged 15 or over, of 11 European countries or regions (all old EU Member States except Denmark, Greece, Ireland, Finland and Great Britain which had to be excluded because of poor response rate, less than 45%). The method of opinion survey was applied using face-to-face interviews. The Energy and Vitality Index (EVI) from the SF-36 questionnaire was use as measure of positive mental health. RESULTS: Overall, there were between-country differences in the gender- and age-adjusted EVI mean scores. In general, poorer mental health was found in women, older age groups, those in poor economic position and those experiencing weak social support. CONCLUSION: Methodological biases cannot be fully excluded, and thus, one has to take the presented results with certain caution, especially when comparing the results from the different countries. On the other hand, the results on the determinants of positive mental health are in concordance with most previous studies. PMID- 16042764 TI - Nonparametric tests for differential gene expression and interaction effects in multi-factorial microarray experiments. AB - BACKGROUND: Numerous nonparametric approaches have been proposed in literature to detect differential gene expression in the setting of two user-defined groups. However, there is a lack of nonparametric procedures to analyze microarray data with multiple factors attributing to the gene expression. Furthermore, incorporating interaction effects in the analysis of microarray data has long been of great interest to biological scientists, little of which has been investigated in the nonparametric framework. RESULTS: In this paper, we propose a set of nonparametric tests to detect treatment effects, clinical covariate effects, and interaction effects for multifactorial microarray data. When the distribution of expression data is skewed or heavy-tailed, the rank tests are substantially more powerful than the competing parametric F tests. On the other hand, in the case of light or medium-tailed distributions, the rank tests appear to be marginally less powerful than the parametric competitors. CONCLUSION: The proposed rank tests enable us to detect differential gene expression and establish interaction effects for microarray data with various non-normally distributed expression measurements across genome. In the presence of outliers, they are advantageous alternative approaches to the existing parametric F tests due to the robustness feature. PMID- 16042765 TI - Medicines and vaccines for the world's poorest: is there any prospect for public private cooperation? AB - This paper reviews the current status of the global pharmaceutical industry and its research and development focus in the context of the health care needs of the developing world. It will consider the attempts to improve access to critical drugs and vaccines, and increase the research effort directed at key public health priorities in the developing world. In particular, it will consider prospects for public-private collaboration. The challenges and opportunities in such public-private partnerships will be discussed briefly along with a look at factors that may be key to success. Much of the focus is on HIV/AIDS where the debate on the optimal balance between intellectual property rights (IPR) and human rights to life and health has been very public and emotive. PMID- 16042766 TI - Assessing influenza-related mortality: comment on Zucs et al. PMID- 16042767 TI - Radiation induced osteogenic sarcoma of the maxilla. AB - BACKGROUND: Radiation induced sarcoma arise as a long term complication of radiation treatment for other benign or malignant conditions. They are of very rare occurrence in jaw bones and are even rarer in maxilla. CASE PRESENTATION: Here we report a case of radiation induced sarcoma in a patient treated for squamous cell carcinoma of buccal mucosa with radiation who developed osteosarcoma of maxillary bone after six years. The patient was treated successfully with surgery. CONCLUSION: What should be the best treatment of radiation induced sarcoma is still debatable; however, surgery offers the best chance of cure. Role of reradiation and adjuvant chemotherapy needs to be further evaluated. PMID- 16042768 TI - Risk of cardio-respiratory abnormalities in preterm infants placed in car seats: a cross-sectional study. AB - BACKGROUND: Little is known about the factors that predispose to the occurrence and severity of cardio-respiratory symptoms during the placement of a prematurely born infant in a car seat. The impact of gestational age, weight at discharge and infant's pre-existing cardio-respiratory status (in the supine position) on cardio-respiratory function during pre-discharge testing in a car seat (semi upright position) has not been investigated. METHODS: The cardio-respiratory function of 42 preterm neonates with gestational age 24 to 35 weeks and discharge weight 1790 to 2570 grams were monitored for 45 minutes before, during, and after placement in a car seat. The occurrence of periodic breathing, apnea, bradycardia, or decreased oxygen saturation (SaO2) was analyzed. RESULTS: Prior to the car seat testing, 15 (35.7%) infants displayed one or more abnormalities of cardio-respiratory function. During the car seat testing, 25 (59.6%) infants had periodic breathing, 33 (78.2%) had oxygen saturation <90%, 14 (33.3%) had bradycardia less than 80 beats per minute, and 35 (83.3%) had a combination of these symptoms. Infants, both with and without pre-existing cardio-respiratory abnormalities, had an almost equal probability (80% vs. 83.3%) for the development of cardio-respiratory symptoms during placement in the car seat. Weight at discharge ([less than or equal to] 2,000 grams) but not the gestational age (<28 weeks or [greater than or equal to] 28<37 weeks), was associated with either increased episodes of oxygen desaturation or the combination of cardio respiratory symptoms that were seen during the placement of these infants in the car seat. Repositioning from the car seat to the supine position showed normalization of cardio-respiratory function in the majority (83%) of the tested infants. None of the tested clinical factors were associated with the severity of the cardio-respiratory symptoms. CONCLUSION: Pre-discharge testing of the cardio respiratory function of preterm infants during placement in a car seat is important for the prevention of cardio-respiratory symptoms during their transportation. However, the high risk for developing cardio-respiratory symptoms will require the consideration of an alternative mode of safe home transportation for preterm infants; especially those with a discharge weight less than 2,000 grams. PMID- 16042769 TI - An encoded N-terminal extension results in low levels of heterologous protein production in Escherichia coli. AB - BACKGROUND: The tdk gene (encoding deoxythymidine kinase) of the gamma proteobacterium Xenorhabdus nematophila has two potential translation start sites. The promoter-distal start site was predicted to be functional based on amino acid sequence alignment with closely related Tdk proteins. However, to experimentally determine if either of the two possible start codons allows production of a functional Tdk, we expressed the "long-form" (using the promoter proximal start codon) and "short-form" (using the promoter-distal start codon) X. nematophila tdk genes from the T7 promoter of the pET-28a(+) vector. We assessed Tdk production and activity using a functional assay in an Escherichia coli tdk mutant, which, since it lacks functional Tdk, is able to grow in 5 fluorodeoxyuridine (FUdR)-containing medium. RESULTS: Short-form Tdk complemented the E. coli tdk mutant strain, resulting in FUdR sensitivity of the strain. However, the E. coli tdk mutant expressing the long form of tdk remained FUdR resistant, indicating it did not have a functional deoxythymidine kinase enzyme. We report that long-form Tdk is at least 13-fold less abundant than short-form Tdk, the limited protein produced was as stable as short-form Tdk and the long form transcript was 1.7-fold less abundant than short-form transcript. Additionally, we report that the long-form extension was sufficient to decrease heterologous production of a different X. nematophila protein, NilC. CONCLUSION: We conclude that the difference in the FUdR growth phenotype between the E. coli tdk mutant carrying the long-or short-form X. nematophila tdk is due to a difference in Tdk levels. The lower long-form protein level does not result from protein instability, but instead from reduced transcript levels possibly combined with reduced translation efficiency. Because the observed effect of the encoded N terminal extension is not specific to Tdk production and can be overcome with induction of gene expression, these results may have particular relevance to researchers attempting to limit production of toxic proteins under non-inducing conditions. PMID- 16042770 TI - Evidence that 17alpha-estradiol is biologically active in the uterine tissue: antiuterotonic and antiuterotrophic action. AB - BACKGROUND: 17alpha-Estradiol has been considered as the hormonally inactive isomer of 17beta-estradiol. Recently, nongenomic (smooth muscle relaxation) and genomic (light estrogenic activity) effects of 17alpha-estradiol have been reported, but no reports have yet determined its possible antiestrogenic activity. Therefore, this study investigated: the nongenomic action of 17alpha estradiol on uterine contractile activity and its potential agonist-antagonist activity on uterine growth. METHODS: Uterine rings from rats were isometrically recorded. Different concentrations (0.2-200 microM) of 17alpha-estradiol were tested on spontaneous contraction and equimolarly compared with 17beta-estradiol. To examine the mechanism of 17alpha-estradiol action, its effect was studied in presence of beta2-antagonist (propranolol), antiestrogens (tamoxifen and ICI 182,780) or inhibitors of protein synthesis (cycloheximide) and transcription (actinomycin D). Moreover, contractions induced by high potassium (KCl) solution or calcium in depolarized tissues by KCl-calcium free solution were exposed to 17alpha-estradiol. Collaterally, we performed an uterotrophic assay in adult ovariectomized rats measuring the uterine wet weight. The administration for three days of 0.3 microM/day/Kg 17beta-estradiol was equimolarly compared with the response produced by 17alpha-estradiol. Antiuterotrophic activity was assayed by administration of 0.3 microM/day/Kg 17beta-estradiol and various doses ratios (1:1, 1:3, 1:5, and 1:100) of 17alpha-estradiol. RESULTS: The estradiol isomers elicited an immediate relaxation, concentration-dependent and reversible on spontaneous contraction. 17alpha-Estradiol presented lower potency than 17beta estradiol although it did not antagonize 17beta-estradiol-induced relaxation. Relaxation to 17alpha-estradiol was not inhibited by propranolol, tamoxifen, ICI 182,780, cycloheximide or actinomycin D. The KCl contractions were also sensitive to 17alpha-estradiol-induced relaxation and calcium contractions in depolarized tissues were markedly prevented by 17alpha-estradiol, implying a reduction of extracellular calcium influx through voltage-operated calcium channels (VOCCs). Uterotrophic assay detected significant increase in uterine weight using 17alpha estradiol, which was significantly minor as compared with 17beta-estradiol. 17alpha-Estradiol, at all doses ratios, significantly antagonized the hypertrophic response of 17beta-estradiol. CONCLUSION: 17alpha-Estradiol induces a relaxing effect, which may be independent of the classical estrogen receptor, nongenomic action, apparently mediated by inactivation of VOCCs. 17alpha Estradiol is also a weak estrogen agonist (uterotrophic response); likewise, 17alpha-estradiol may act as an antiestrogen (antiuterotrophic response). The overall data document a nongenomic relaxing action and a novel antiestrogenic action of 17alpha-estradiol, which are relevant in estrogen-mediated uterine physiology. PMID- 16042771 TI - Exhaled 8-isoprostane in childhood asthma. AB - BACKGROUND: Exhaled breath condensate (EBC) is a non-invasive method to assess airway inflammation and oxidative stress and may be useful in the assessment of childhood asthma. METHODS: Exhaled 8-isoprostane, a stable marker of oxidative stress, was measured in EBC, in children (5-17 years) with asthma (13 steroid naive and 12 inhaled steroid-treated) and 11 healthy control. RESULTS: Mean exhaled 8-isoprostane concentration was significantly elevated in steroid-naive asthmatic children compared to healthy children 9.3 (SEM 1.7) vs. 3.8 (0.6) pg/ml, p < 0.01. Children on inhaled steroids also had significantly higher 8 isoprostane levels than those of normal subjects 6.7 (0.7) vs. 3.8 (0.6) pg/ml, p < 0.01. Steroid-naive asthmatics had higher exhaled nitric oxide (eNO) than those of controls 28.5 (4.7) vs. 12.6 (1.5) ppb, p < 0.01. eNO in steroid-treated asthmatics was similar to control subjects 27.5(8.8) vs. 12.6(1.5) ppb. Exhaled 8 isoprostane did not correlate with duration of asthma, dose of inhaled steroids or eNO. CONCLUSION: We conclude that 8-isoprostane is elevated in asthmatic children, indicating increased oxidative stress, and that this does not appear to be normalized by inhaled steroid therapy. This suggests that 8-isoprostane is a useful non-invasive measurement of oxidative stress in children and that antioxidant therapy may be useful in the future. PMID- 16042772 TI - Zollinger-Ellison syndrome associated with neurofibromatosis type 1: a case report. AB - BACKGROUND: Neurofibromatosis type 1 is an autosomal dominant neurocutaneous disorder with characteristic features of skin and central nervous system involvement. Gastrointestinal involvement is rare, but the risk of malignancy development is considerable. Zollinger-Ellison syndrome is caused by gastrin secreting tumors called gastrinomas. Correct diagnosis is often difficult, and curative treatment can only be achieved surgically. CASE PRESENTATION: A 41-year old female affected by neurofibromatosis type 1 presented with a history of recurrent epigastric soreness, diarrhea, and relapsing chronic duodenal ulcer. Her serum fasting gastrin level was over 1000 pg/mL. An abdominal CT scan revealed a 3 x 2-cm, well-enhanced mass adjacent to the duodenal loop. She was not associated with multiple endocrine neoplasia type 1. Operative resection was performed and gastrinoma was diagnosed by immunohistochemical staining. The serum gastrin level decreased to 99.1 pg/mL after surgery, and symptoms and endoscopic findings completely resolved without recurrences. CONCLUSION: Gastrinoma is difficult to detect even in the general population, and hence symptoms such as recurrent idiopathic peptic ulcer and diarrhea in neurofibromatosis type 1 patients should be accounted for as possibly contributing to Zollinger-Ellison syndrome. PMID- 16042773 TI - Antenatal screening for Group B Streptococcus: a diagnostic cohort study. AB - BACKGROUND: A range of strategies have been adopted to prevent early onset Group B Streptococcal (EOGBS) sepsis, as a consequence of Group B Streptococcal (GBS) vertically acquired infection. This study was designed to provide a scientific basis for optimum timing and method of GBS screening in an Australian setting, to determine whether screening for GBS infection at 35-37 weeks gestation has better predictive values for colonisation at birth than screening at 31-33 weeks, to examine the test characteristics of a risk factor strategy and to determine the test characteristics of low vaginal swabs alone compared with a combination of perianal plus low vaginal swabs per colonisation during labour. METHODS: Consented women received vaginal and perianal swabs at 31-33 weeks gestation, 35 38 weeks gestation and during labour. Swabs were cultured on layered horse blood agar and inoculated into selective broth prior to analysis. Test characteristics were calculated with exact confidence intervals for a high risk strategy and for antenatal screening at 31-33 and 35-37 weeks gestation for vaginal cultures alone, perianal cultures alone and combined low vaginal and perianal cultures. RESULTS: The high risk strategy was not informative in predicting GBS status during labour. There is an unequivocal benefit for the identification of women colonised with GBS during labour associated with delaying screening until 36 weeks however the results for method of screening were less definitive with no clear advantage in using a combined low vaginal and perianal swabbing regimen over the use of a low vaginal swab alone. CONCLUSION: This study can contribute to the development of prevention strategies in that it provides clear evidence for optimal timing of swabs. The addition of a perianal swab does not confer clear benefit. The quantification of advantages and disadvantages provided in this study will facilitate communication with clinicians and pregnant women alike. PMID- 16042774 TI - Expression profiles of hydrophobic surfactant proteins in children with diffuse chronic lung disease. AB - BACKGROUND: Abnormalities of the intracellular metabolism of the hydrophobic surfactant proteins SP-B and SP-C and their precursors may be causally linked to chronic childhood diffuse lung diseases. The profile of these proteins in the alveolar space is unknown in such subjects. METHODS: We analyzed bronchoalveolar lavage fluid by Western blotting for SP-B, SP-C and their proforms in children with pulmonary alveolar proteinosis (PAP, n = 15), children with no SP-B (n = 6), children with chronic respiratory distress of unknown cause (cRD, n = 7), in comparison to children without lung disease (n = 15) or chronic obstructive bronchitis (n = 19). RESULTS: Pro-SP-B of 25-26 kD was commonly abundant in all groups of subjects, suggesting that their presence is not of diagnostic value for processing defects. In contrast, pro-SP-B peptides cleaved off during intracellular processing of SP-B and smaller than 19-21 kD, were exclusively found in PAP and cRD. In 4 of 6 children with no SP-B, mutations of SFTPB or SPTPC genes were found. Pro-SP-C forms were identified at very low frequency. Their presence was clearly, but not exclusively associated with mutations of the SFTPB and SPTPC genes, impeding their usage as candidates for diagnostic screening. CONCLUSION: Immuno-analysis of the hydrophobic surfactant proteins and their precursor forms in bronchoalveolar lavage is minimally invasive and can give valuable clues for the involvement of processing abnormalities in pediatric pulmonary disorders. PMID- 16042775 TI - Methodology and measurement properties of health-related quality of life instruments: a prospective study of patients undergoing breast reduction surgery. AB - BACKGROUND: Breast hypertrophy is associated with clinically important morbidity. A prospective study was conducted to assess the change in health-related quality of life (HRQL) following breast reduction mammoplasty. This paper describes the measurement properties of each of the HRQL questionnaires used. METHODS: The reliability, responsiveness, and the construct validity of each HRQL instrument (the Health Utilities Index Mark 2 (HUI2) and Mark 3 (HUI3) and the Breast Reduction Assessment Value and Outcomes (BRAVO) instruments) were assessed. The BRAVO instruments are a set of separate instruments including the Short Form-36 (SF-36), the Multidimensional Body Self Relations Questionnaire Appearance Assessment (MBSRQ-AS), and the Breast Related Symptoms Questionnaire (BRSQ). RESULTS: The HUI2, the HUI3, the MBSRQ-AS, and the breast severity symptom (BSS) score from the BRSQ all demonstrated good test-retest reliability. The SF-36 physical component summary, the MBSRQ-AS, and the BSS score demonstrated high responsiveness. The SF-36 mental component summary and the HUI3 had a moderate effect size and the HUI2 had a small effect size. All of the changes in scales are correlated in the same direction except for the SF-36 physical component summary and the SF-36 mental component summary. CONCLUSION: All four instruments were found to be reliable and responsive. These instruments can be used in similar clinical settings to evaluate the change in patients' HRQL. PMID- 16042776 TI - Inhibition of chemokine expression in rat inflamed paws by systemic use of the antihyperalgesic oxidized ATP. AB - BACKGROUND: We previously showed that local use of periodate oxidized ATP (oATP, a selective inhibitor of P2X7 receptors for ATP) in rat paw treated with Freund's adjuvant induced a significant reduction of hyperalgesia Herein we investigate the role of oATP, in the rat paws inflamed by carrageenan, which mimics acute inflammation in humans. RESULTS: Local, oral or intravenous administration of a single dose of oATP significantly reduced thermal hyperalgesia in hind paws of rats for 24 hours, and such effect was greater than that induced by diclofenac or indomethacin. Following oATP treatment, the expression of the pro-inflammatory chemokines interferon-gamma-inducible protein-10 (IP-10), mon ocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) within the inflamed tissues markedly decreased on vessels and infiltrated cells. In parallel, the immunohistochemical findings showed an impairment, with respect to the untreated rats, in P2X7 expression, mainly on nerves and vessels close to the site of inflammation. Finally, oATP treatment significantly reduced the presence of infiltrating inflammatory macrophages in the paw tissue. CONCLUSION: Taken together these results clearly show that oATP reduces carrageenan-induced inflammation in rats. PMID- 16042777 TI - Psychometric properties of the CDC Symptom Inventory for assessment of chronic fatigue syndrome. AB - OBJECTIVES: Validated or standardized self-report questionnaires used in research studies and clinical evaluation of chronic fatigue syndrome (CFS) generally focus on the assessment of fatigue. There are relatively few published questionnaires that evaluate case defining and other accompanying symptoms in CFS. This paper introduces the self-report CDC CFS Symptom Inventory and analyzes its psychometric properties. METHODS: One hundred sixty-four subjects (with CFS, other fatiguing illnesses and non fatigued controls) identified from the general population of Wichita, Kansas were enrolled. Evaluation included a physical examination, a standardized psychiatric interview, three previously validated self-report questionnaires measuring fatigue and illness impact (Medical Outcomes Survey Short-Form-36 [MOS SF-36], Multidimensional Fatigue Inventory [MFI], Chalder Fatigue Scale), and the CDC CFS Symptom Inventory. Based on theoretical assumptions and statistical analyses, we developed several different Symptom Inventory scores and evaluated them on their ability to differentiate between participants with CFS and non-fatigued controls. RESULTS: The Symptom Inventory had good internal consistency and excellent convergent validity. A Total score (all symptoms), Case Definition score (CFS case defining symptoms) and Short Form score (6 symptoms with minimal correlation) differentiated CFS cases from controls. Furthermore, both the Case Definition and Short Form scores distinguished people with CFS from fatigued subjects who did not meet criteria for CFS. CONCLUSION: The Symptom Inventory appears to be a reliable and valid instrument to assess symptoms that accompany CFS. It is a positive addition to existing instruments measuring fatigue because it allows other dimensions of the illness to be assessed. Further research is needed to confirm and replicate the current findings in a normative population. PMID- 16042778 TI - Utility of a thematic network in primary health care: a controlled interventional study in a rural area. AB - BACKGROUND: UniNet is an Internet-based thematic network for a virtual community of users (VCU). It supports a virtual multidisciplinary community for physicians, focused on the improvement of clinical practice. This is a study of the effects of a thematic network such as UniNet on primary care medicine in a rural area, specifically as a platform of communication between specialists at the hospital and doctors in the rural area. METHODS: In order to study the effects of a thematic network such as UniNet on primary care medicine in a rural area, we designed an interventional study that included a control group. The measurements included the number of patient displacements due to disease, number of patient hospital stays and the number of prescriptions of drugs of low therapeutic utility and generic drug prescriptions by doctors. These data were analysed and compared with those of the control center. RESULTS: Our study showed positive changes in medical practice, reflected in the improvement of the evaluated parameters in the rural health area where the interventional study was carried out, compared with the control area. We discuss the strengths and weaknesses of UniNet as a potential medium to improve the quality of medical care in rural areas. CONCLUSION: The rural doctors had an effective, useful, user-friendly and cheap source of medical information that may have contributed to the improvement observed in the medical quality indices. PMID- 16042779 TI - Rank-invariant resampling based estimation of false discovery rate for analysis of small sample microarray data. AB - BACKGROUND: The evaluation of statistical significance has become a critical process in identifying differentially expressed genes in microarray studies. Classical p-value adjustment methods for multiple comparisons such as family-wise error rate (FWER) have been found to be too conservative in analyzing large screening microarray data, and the False Discovery Rate (FDR), the expected proportion of false positives among all positives, has been recently suggested as an alternative for controlling false positives. Several statistical approaches have been used to estimate and control FDR, but these may not provide reliable FDR estimation when applied to microarray data sets with a small number of replicates. RESULTS: We propose a rank-invariant resampling (RIR) based approach to FDR evaluation. Our proposed method generates a biologically relevant null distribution, which maintains similar variability to observed microarray data. We compare the performance of our RIR-based FDR estimation with that of four other popular methods. Our approach outperforms the other methods both in simulated and real microarray data. CONCLUSION: We found that the SAM's random shuffling and SPLOSH approaches were liberal and the other two theoretical methods were too conservative while our RIR approach provided more accurate FDR estimation than the other approaches. PMID- 16042780 TI - Creating an "enabling environment" for taking insecticide treated nets to national scale: the Tanzanian experience. AB - INTRODUCTION: Malaria is the largest cause of health services attendance, hospital admissions and child deaths in Tanzania. At the Abuja Summit in April 2000 Tanzania committed itself to protect 60% of its population at high risk of malaria by 2005. The country is, therefore, determined to ensure that sustainable malaria control using insecticide-treated nets is carried out on a national scale. CASE DESCRIPTION: Tanzania has been involved for two decades in the research process for developing insecticide-treated nets as a malaria control tool, from testing insecticides and net types, to assessing their efficacy and effectiveness, and exploring new ways of distribution. Since 2000, the emphasis has changed from a project approach to that of a concerted multi-stakeholder action for taking insecticide-treated nets to national scale (NATNETS). This means creating conditions that make insecticide-treated nets accessible and affordable to all those at risk of malaria in the country. This paper describes Tanzania's experience in (1) creating an enabling environment for insecticide treated nets scale-up, (2) promoting the development of a commercial sector for insecticide-treated nets, and (3) targeting pregnant women with highly subsidized insecticide-treated nets through a national voucher scheme. As a result, nearly 2 million insecticide-treated nets and 2.2 million re-treatment kits were distributed in 2004. CONCLUSION: National upscaling of insecticide-treated nets is possible when the programme is well designed, coordinated and supported by committed stakeholders; the Abuja target of protecting 60% of those at high risk is feasible, even for large endemic countries. PMID- 16042781 TI - Dynamin-dependent NMDAR endocytosis during LTD and its dependence on synaptic state. AB - BACKGROUND: The N-methyl-D-aspartate (NMDA)-type glutamate receptor expressed at excitatory glutamatergic synapses is required for learning and memory and is critical for normal brain function. At a cellular level, this receptor plays a pivotal role in triggering and controlling synaptic plasticity. While it has been long recognized that this receptor plays a regulatory role, it was considered by many to be itself immune to synaptic activity-induced plasticity. More recently, we and others have shown that NMDA receptor-mediated synaptic responses can be subject to activity-dependent depression. RESULTS: Here we show that depression of synaptic transmission mediated by NMDA receptors displays a state-dependence in its plasticity; NMDA receptors are resistant to activity-induced changes at silent and recently-silent synapses. Once synapses transition to the active state however, NMDA receptors become fully 'plastic'. This state-dependence is identical to that shown by the alpha-amino-3-hydroxy-5-methylisoxazole-4 propionic acid (AMPA) receptor. Furthermore, the down-regulation of NMDAR mediated responses during synaptic depression is prevented by disruption of dynamin-dependent endocytosis. CONCLUSION: NMDA receptor-mediated synaptic responses are plastic in a state-dependent manner. Depending on the plasticity state in which a synapse currently resides, NMDA receptors will either be available or unavailable for down-regulation. The mechanism underlying the down regulation of NMDA receptor-mediated synaptic responses is endocytosis of the NMDA receptor. Other potential mechanisms, such as receptor diffusion along the plane of the membrane, or changes in the activity of the channel are not supported. The mechanisms of AMPA receptor and NMDA receptor endocytosis appear to be tightly coupled, as both are either available or unavailable for endocytosis in the same synaptic states. Endocytosis of NMDA receptors would serve as a potent mechanism for metaplasticity. Such state-dependent regulation of NMDAR endocytosis will provide fundamental control over downstream NMDA receptor-dependent plasticity of neuronal circuitry. PMID- 16042782 TI - Children's acceptance of milk with xylitol or sorbitol for dental caries prevention. AB - BACKGROUND: Xylitol, a polyol sugar, has been shown to reduce dental caries when mixed with food or chewing gum. This study examines the taste acceptability of xylitol in milk as a first step toward measuring the effectiveness of xylitol in milk for the reduction of dental caries in a public health program. METHODS: Three different types of milk (Ultra High Temperature (UHT), powder and evaporated) were tested for acceptability by 75 Peruvian children (25 per milk group, ages 4 to 7 years). Each group evaluated xylitol and sorbitol in one type of milk. In the first phase, each child was presented with a tray of four plastic cups containing 50 ml of milk with 0.021 g/ml xylitol, 0.042 g/ml xylitol, 0.042 g/ml sorbitol or no sugar. Each child was asked to taste the samples in a self selected order. After tasting each sample, the child placed the milk cup in front of one of three cartoon faces (smile, frown or neutral) representing the child's response to the taste of each sample. In the second phase, the child was asked to rank order the milk samples within each category (smile, frown or neutral). Ranks within categories were then combined to obtain a rank ordering for all the test samples. RESULTS: The ranking from best to worst for the samples across categories (UHT, powder, evaporated) was xylitol (0.0.042 g/ml), sorbitol (0.042 g/ml), xylitol (0.021 g/ml) and milk alone (Friedman's ANOVA). Xylitol and sorbitol were preferred over milk alone, and xylitol (0.042 g/ml) was preferred to sorbitol (0.042 g/ml)(p < .05 sign test). CONCLUSION: Milk sweetened with xylitol is well accepted by Peruvian children ages 4-7 years. PMID- 16042783 TI - Comparing independent microarray studies: the case of human embryonic stem cells. AB - BACKGROUND: Microarray studies of the same phenomenon in different labs often appear at variance because the published lists of regulated transcripts have disproportionately small intersections. We demonstrate that comparing studies by intersecting lists in this manner is methodologically flawed by reanalyzing three studies of the molecular signature of "stemness" in human embryonic stem cells. There are only 7 genes common to all three published lists, suggesting disagreement. RESULTS: Carefully reanalyzing all three together from the raw data we detect 111 genes upregulated and 95 downregulated in all three studies. The upregulated list was subject to rtRTPCR analysis and 75% of the genes were confirmed. CONCLUSION: Our findings show that the three studies have a substantial core of common genes, which is missed if only the published lists are examined. Combined analysis of multiple experiments can be a powerful way to distil coherent conclusions. PMID- 16042784 TI - A role for the Tec family kinase ITK in regulating SEB-induced interleukin-2 production in vivo via c-jun phosphorylation. AB - BACKGROUND: Exposure to Staphylococcal Enterotoxin B (SEB), a bacterial superantigen secreted by the Gram-positive bacteria Staphyloccocus aureus, results in the expansion and eventual clonal deletion and anergy of Vbeta8+ T cells, as well as massive cytokine release, including Interleukin-2 (IL-2). This IL-2 is rapidly secreted following exposure to SEB and may contribute to the symptoms seen following exposure to this bacterial toxin. The Tec family kinase ITK has been shown to be important for the production of IL-2 by T cells stimulated in vitro and may represent a good target for blocking the production of this cytokine in vivo. In order to determine if ITK represents such a target, mice lacking ITK were analyzed for their response to SEB exposure. RESULTS: It was found that T cells from mice lacking ITK exhibited significantly reduced proliferative responses to SEB exposure in vitro, as well as in vivo. Examination of IL-2 production revealed that ITK null mice produced reduced levels of this cytokine in vitro, and more dramatically, in vivo. In vivo analysis of c-jun phosphorylation, previously shown to be critical for regulating IL-2 production, revealed that this pathway was specifically activated in SEB reactive Vbeta8+ (but not non-reactive Vbeta6+) T cells from WT mice, but not in Vbeta8+ T cells from ITK null mice. However, toxicity analysis indicated that both WT and ITK null animals were similarly affected by SEB exposure. CONCLUSION: These data show that ITK is required for IL-2 production induced by SEB in vivo, and may regulate signals leading IL-2 production, in part by regulating phosphorylation of c-jun. The data also suggest that perturbing T cell activation pathways leading to IL-2 does not necessarily lead to improved responses to SEB toxicity. PMID- 16042785 TI - Application of Affymetrix array and Massively Parallel Signature Sequencing for identification of genes involved in prostate cancer progression. AB - BACKGROUND: Affymetrix GeneChip Array and Massively Parallel Signature Sequencing (MPSS) are two high throughput methodologies used to profile transcriptomes. Each method has certain strengths and weaknesses; however, no comparison has been made between the data derived from Affymetrix arrays and MPSS. In this study, two lineage-related prostate cancer cell lines, LNCaP and C4-2, were used for transcriptome analysis with the aim of identifying genes associated with prostate cancer progression. METHODS: Affymetrix GeneChip array and MPSS analyses were performed. Data was analyzed with GeneSpring 6.2 and in-house perl scripts. Expression array results were verified with RT-PCR. RESULTS: Comparison of the data revealed that both technologies detected genes the other did not. In LNCaP, 3,180 genes were only detected by Affymetrix and 1,169 genes were only detected by MPSS. Similarly, in C4-2, 4,121 genes were only detected by Affymetrix and 1,014 genes were only detected by MPSS. Analysis of the combined transcriptomes identified 66 genes unique to LNCaP cells and 33 genes unique to C4-2 cells. Expression analysis of these genes in prostate cancer specimens showed CA1 to be highly expressed in bone metastasis but not expressed in primary tumor and EPHA7 to be expressed in normal prostate and primary tumor but not bone metastasis. CONCLUSION: Our data indicates that transcriptome profiling with a single methodology will not fully assess the expression of all genes in a cell line. A combination of transcription profiling technologies such as DNA array and MPSS provides a more robust means to assess the expression profile of an RNA sample. Finally, genes that were differentially expressed in cell lines were also differentially expressed in primary prostate cancer and its metastases. PMID- 16042786 TI - Phase I dose-escalating study of docetaxel in combination with 5-day continuous infusion of 5-fluorouracil in patients with advanced gastric cancer. AB - BACKGROUND: Published data suggests that docetaxel combined with 5-fluorouracil (5-FU) may have synergistic activity in treating advanced gastric cancer. We performed a phase I study of docetaxel and 5-FU to determine the maximum tolerated dose (MTD), the recommended dose for phase II studies, and the safety of this combination. METHODS: Eligible patients had recurrent and/or metastatic advanced gastric cancer with normal cardiac, renal and hepatic function. Traditional phase I methodology was employed in assessing dose-limiting toxicity (DLT) and MTD. On day 1 every 3 weeks, docetaxel 75 mg/m2 (fixed dose) was infused over 1-h, followed immediately by 5-FU as a 5-day continuous infusion. RESULTS: Dose escalation schema was as follows: dose level (DL) 1 (5-FU 250 mg/m2/day), 2 (500), 3 (750), and 4 (1000). Three patients were enrolled on DL1, without DLT. On DL2, 1 DLT (grade 3 stomatitis) was developed in first 3 patients, and this cohort was expanded to 6 patients. Three patients had been enrolled on DL3. Because two out of 3 patients had DLTs, the MTD was reached at DL3. CONCLUSION: The recommended phase II dose of this combination is 75 mg/m2 docetaxel on day 1 immediately followed by a 5-day continuous infusion of 5-FU 500 mg/m2/day. PMID- 16042787 TI - PDZ domain-binding motif of human T-cell leukemia virus type 1 Tax oncoprotein is essential for the interleukin 2 independent growth induction of a T-cell line. AB - BACKGROUND: Human T-cell leukemia virus type 1 (HTLV-1) is the etiologic agent of adult T-cell leukemia (ATL), whereas HTLV type 2 (HTLV-2), is not associated with ATL or any other leukemia. HTLV-1 encodes the transforming gene tax1, whose expression in an interleukin (IL)-2-dependent T-cell line (CTLL-2) induces IL-2 independent growth. RESULTS: In this study, we demonstrated that IL-2-independent growth induction by Tax1 was abrogated by mutations of the PDZ domain-binding motif (PBM) at the Tax1 C-terminus. HTLV-2 Tax2, which shares 75% amino acid identity with Tax1 but does not have a PBM, was not able to induce IL-2 independent growth of CTLL-2. CONCLUSION: Our results suggest that Tax1, through interaction with PDZ domain protein(s) induces IL-2-independent growth, which may be a factor in multi-step leukemogenesis caused by HTLV-1. PMID- 16042789 TI - A simplified search strategy for identifying randomised controlled trials for systematic reviews of health care interventions: a comparison with more exhaustive strategies. AB - BACKGROUND: It is generally believed that exhaustive searches of bibliographic databases are needed for systematic reviews of health care interventions. The CENTRAL database of controlled trials (RCTs) has been built up by exhaustive searching. The CONSORT statement aims to encourage better reporting, and hence indexing, of RCTs. Our aim was to assess whether developments in the CENTRAL database, and the CONSORT statement, mean that a simplified RCT search strategy for identifying RCTs now suffices for systematic reviews of health care interventions. METHODS: RCTs used in the Cochrane reviews were identified. A brief RCT search strategy (BRSS), consisting of a search of CENTRAL, and then for variants of the word random across all fields (random$.af.) in MEDLINE and EMBASE, was devised and run. Any trials included in the meta-analyses, but missed by the BRSS, were identified. The meta-analyses were then re-run, with and without the missed RCTs, and the differences quantified. The proportion of trials with variants of the word random in the title or abstract was calculated for each year. The number of RCTs retrieved by searching with "random$.af." was compared to the highly sensitive search strategy (HSSS). RESULTS: The BRSS had a sensitivity of 94%. It found all journal RCTs in 47 of the 57 reviews. The missing RCTs made some significant differences to a small proportion of the total outcomes in only five reviews, but no important differences in conclusions resulted. In the post-CONSORT years, 1997-2003, the percentage of RCTs with random in the title or abstract was 85%, a mean increase of 17% compared to the seven years pre-CONSORT (95% CI, 8.3% to 25.9%). The search using random$.af. reduced the MEDLINE retrieval by 84%, compared to the HSSS, thereby reducing the workload of checking retrievals. CONCLUSION: A brief RCT search strategy is now sufficient to locate RCTs for systematic reviews in most cases. Exhaustive searching is no longer cost-effective, because in effect it has already been done for CENTRAL. PMID- 16042788 TI - Prediction of the general transcription factors associated with RNA polymerase II in Plasmodium falciparum: conserved features and differences relative to other eukaryotes. AB - BACKGROUND: To date, only a few transcription factors have been identified in the genome of the parasite Plasmodium falciparum, the causative agent of malaria. Moreover, no detailed molecular analysis of its basal transcription machinery, which is otherwise well-conserved in the crown group of eukaryotes, has yet been reported. In this study, we have used a combination of sensitive sequence analysis methods to predict the existence of several parasite encoded general transcription factors associated with RNA polymerase II. RESULTS: Several orthologs of general transcription factors associated with RNA polymerase II can be predicted among the hypothetical proteins of the P. falciparum genome using the two-dimensional Hydrophobic Cluster Analysis (HCA) together with profile based search methods (PSI-BLAST). These predicted orthologous genes encoding putative transcription factors include the large subunit of TFIIA and two candidates for its small subunit, the TFIIE beta-subunit, which would associate with the previously known TFIIE alpha-subunit, the TFIIF beta-subunit, as well as the p62/TFB1 subunit of the TFIIH core. Within TFIID, the putative orthologs of TAF1, TAF2, TAF7 and TAF10 were also predicted. However, no candidates for TAFs with classical histone fold domain (HFD) were found, suggesting an unusual architecture of TFIID complex of RNA polymerase II in the parasite. CONCLUSION: Taken together, these results suggest that more general transcription factors may be present in the P. falciparum proteome than initially thought. The prediction of these orthologous general transcription factors opens the way for further studies dealing with transcriptional regulation in P. falciparum. These alternative and sensitive sequence analysis methods can help to identify candidates for other transcriptional regulatory factors in P. falciparum. They will also facilitate the prediction of biological functions for several orphan proteins from other apicomplexan parasites such as Toxoplasma gondii, Cryptosporidium parvum and Eimeria. PMID- 16042790 TI - CD8+ T lymphocytes in lung tissue from patients with idiopathic pulmonary fibrosis. AB - BACKGROUND: Several studies have implicated a role of inflammation in the pathogenesis of lung damage in idiopathic pulmonary fibrosis (IPF). Parenchymal lung damage leads to defects in mechanics and gas exchange and clinically manifests with exertional dyspnea. Investigations of inflammatory cells in IPF have shown that eosinophils, neutrophils and CD8+ TLs may be associated with worse prognosis. We wished to investigate by quantitative immunohistochemistry infiltrating macrophages, neutrophils and T lymphocytes (TLs) subpopulations (CD3+, CD4+ and CD8+) in lung tissue of patients with IPF and their correlation with lung function indices and grade of dyspnoea. METHODS: Surgical biopsies of 12 patients with IPF were immunohistochemically stained with mouse monoclonal antibodies (anti-CD68 for macrophages, anti-elastase for neutrophils, and anti CD3, anti-CD4, anti-CD8 for CD3+TLs, CD4+TLs, and CD8+TLs respectively). The number of positively stained cells was determined by observer-interactive computerized image analysis (SAMBA microscopic image processor). Cell numbers were expressed in percentage of immunopositive nuclear surface in relation to the total nuclear surface of infiltrative cells within the tissue (labeling Index). Correlations were performed between cell numbers and physiological indices [FEV1, FVC, TLC, DLCO, PaO2, PaCO2 and P(A-a)O2)] as well as dyspnoea scores assessed by the Medical Research Council (MRC) scale. RESULTS: Elastase positive cells accounted for the 7.04% +/- 1.1 of total cells, CD68+ cells for the 16.6% +/- 2, CD3+ TLs for the 28.8% +/- 7, CD4+ TLs for the 14.5 +/- 4 and CD8+ TLs for the 13.8 +/- 4. CD8+TLs correlated inversely with FVC % predicted (rs = -0.67, p = 0.01), TLC % predicted (rs = -0.68, p = 0.01), DLCO % predicted (rs = -0.61, p = 0.04), and PaO2 (rs = -0.60, p = 0.04). Positive correlations were found between CD8+TLs and P(A-a)O2 (rs = 0.65, p = 0.02) and CD8+TLs and MRC score (rs = 0.63, p = 0.02). Additionally, CD68+ cells presented negative correlations with both FVC % predicted (rs = -0.80, p = 0.002) and FEV1 % predicted (rs = -0.68, p = 0.01). CONCLUSION: In UIP/IPF tissue infiltrating mononuclear cells and especially CD8+ TLs are associated with the grade of dyspnoea and functional parameters of disease severity implicating that they might play a role in its pathogenesis. PMID- 16042792 TI - Multidrug resistance-associated protein 1 (MRP1) mediated vincristine resistance: effects of N-acetylcysteine and Buthionine sulfoximine. AB - BACKGROUND: Multidrug resistance mediated by the multidrug resistance-associated protein 1 (MRP1) decreases cellular drug accumulation. The exact mechanism of MRP1 involved multidrug resistance has not been clarified yet, though glutathione (GSH) is likely to have a role for the resistance to occur. N-acetylcysteine (NAC) is a pro-glutathione drug. DL-Buthionine (S,R)-sulfoximine (BSO) is an inhibitor of GSH synthesis. The aim of our study was to investigate the effect of NAC and BSO on MRP1-mediated vincristine resistance in Human Embryonic Kidney (HEK293) and its MRP1 transfected 293MRP cells. Human Embryonic Kidney (HEK293) cells were transfected with a plasmid encoding whole MRP1 gene. Both cells were incubated with vincristine in the presence or absence of NAC and/or BSO. The viability of both cells was determined under different incubation conditions. GSH, Glutathione S-Transferase (GST) and glutathione peroxidase (GPx) levels were measured in the cell extracts obtained from both cells incubated with different drugs. RESULTS: N-acetylcysteine increased the resistance of both cells against vincristine and BSO decreased NAC-enhanced MRP1-mediated vincristine resistance, indicating that induction of MRP1-mediated vincristine resistance depends on GSH. Vincristine decreased cellular GSH concentration and increased GPx activity. Glutathione S-Transferase activity was decreased by NAC. CONCLUSION: Our results demonstrate that NAC and BSO have opposite effects in MRP1 mediated vincristine resistance and BSO seems a promising chemotherapy improving agent in MRP1 overexpressing tumor cells. PMID- 16042793 TI - Use of insecticide-treated nets (ITNs) following a malaria education intervention in Piron, Mali: a control trial with systematic allocation of households. AB - BACKGROUND: Insecticide-treated nets (ITNs) reduce malaria morbidity and mortality, but use is limited. A barrier to ITN use may be lack of knowledge regarding malaria transmission and prevention. This study is a controlled trial comparing ITN use and malaria knowledge levels between households in Piron, Mali, undertaken in 2003. METHODS: Households received net impregnation services either with or without antecedent education. The main outcome measure was ITN use, defined as impregnation of at least one of the household's existing bednets with insecticide during the study. Knowledge about malaria and prevention practices was assessed pre- and post- educational intervention. Results were analysed by household and by individual. RESULTS: Forty-nine percent (34/70) of households who received the educational component impregnated their nets in comparison to 35% (22/62) of households who did not (OR = 1.6 CI = 0.8-3.3, P = 0.19). In individual analysis, ITN use was significantly greater in participants who had received the educational intervention (48%) vs. individuals who did not (33%, OR = 1.9, P = 0.012). Knowledge levels about malaria significantly increased for each individual pre- versus post- educational intervention (average change score = 2.13, standard deviation = 1.97, t = -17.78, P < 0.001), although there was no difference found between educational (change score = 2.14) and control groups (change score = 2.12). CONCLUSION: It is possible to educate individuals about malaria and to implement net impregnation services with limited resources. Greater accessibility to net-impregnation services is necessary but not sufficient to increase ITN use. PMID- 16042794 TI - Application of four dyes in gene expression analyses by microarrays. AB - BACKGROUND: DNA microarrays are widely used in gene expression analyses. To increase throughput and minimize costs without reducing gene expression data obtained, we investigated whether four mRNA samples can be analyzed simultaneously by applying four different fluorescent dyes. RESULTS: Following tests for cross-talk of fluorescence signals, Alexa 488, Alexa 594, Cyanine 3 and Cyanine 5 were selected for hybridizations. For self-hybridizations, a single RNA sample was labelled with all dyes and hybridized on commercial cDNA arrays or on in-house spotted oligonucleotide arrays. Correlation coefficients for all combinations of dyes were above 0.9 on the cDNA array. On the oligonucleotide array they were above 0.8, except combinations with Alexa 488, which were approximately 0.5. Standard deviation of expression differences for replicate spots were similar on the cDNA array for all dye combinations, but on the oligonucleotide array combinations with Alexa 488 showed a higher variation. CONCLUSION: In conclusion, the four dyes can be used simultaneously for gene expression experiments on the tested cDNA array, but only three dyes can be used on the tested oligonucleotide array. This was confirmed by hybridizations of control with test samples, as all combinations returned similar numbers of differentially expressed genes with comparable effects on gene expression. PMID- 16042795 TI - A case report: Pavlovian conditioning as a risk factor of heroin 'overdose' death. AB - BACKGROUND: The authors present a case illustrating a mechanism leading directly to death which is not rare but has received little attention. CASE PRESENTATION: The case was evaluated by autopsy, investigation of morphine concentration in the blood, and clinical data. The heroin dose causing the 'overdose' death of a young man who had previously been treated a number of times for heroin addiction did not differ from his dose of the previous day taken in the accustomed circumstances. The accustomed dose taken in a strange environment caused fatal complications because the conditioned tolerance failed to operate. The concentration of morphine in the blood did not exceed the level measured during earlier treatment. CONCLUSION: These results are in line with the data in the literature indicating that morphine concentrations measured in cases of drug related death do not differ substantially from those measured in cases where the outcome is not fatal. A knowledge of the conditioning mechanism can contribute to prevention of fatal cases of a similar type. The harm reduction approach places great stress on preventive intervention based on data related to drug-related death. PMID- 16042796 TI - Psychophysiological effects of a web-based stress management system: a prospective, randomized controlled intervention study of IT and media workers [ISRCTN54254861]. AB - BACKGROUND: The aim of the present study was to assess possible effects on mental and physical well-being and stress-related biological markers of a web-based health promotion tool. METHODS: A randomized, prospectively controlled study was conducted with before and after measurements, involving 303 employees (187 men and 116 women, age 23-64) from four information technology and two media companies. Half of the participants were offered web-based health promotion and stress management training (intervention) lasting for six months. All other participants constituted the reference group. Different biological markers were measured to detect possible physiological changes. RESULTS: After six months the intervention group had improved statistically significantly compared to the reference group on ratings of ability to manage stress, sleep quality, mental energy, concentration ability and social support. The anabolic hormone dehydroepiandosterone sulphate (DHEA-S) decreased significantly in the reference group as compared to unchanged levels in the intervention group. Neuropeptide Y (NPY) increased significantly in the intervention group compared to the reference group. Chromogranin A (CgA) decreased significantly in the intervention group as compared to the reference group. Tumour necrosis factor alpha (TNFalpha) decreased significantly in the reference group compared to the intervention group. Logistic regression analysis revealed that group (intervention vs. reference) remained a significant factor in five out of nine predictive models. CONCLUSION: The results indicate that an automatic web-based system might have short-term beneficial physiological and psychological effects and thus might be an opportunity in counteracting some clinically relevant and common stress and health issues of today. PMID- 16042797 TI - The effect of prior statin use on 30-day mortality for patients hospitalized with community-acquired pneumonia. AB - BACKGROUND: Recent studies suggest that HMG-CoA reductase inhibitors ("statins") may have beneficial effects for patients at risk for some types of infections. We examined the effect of prior outpatient use of statins on mortality for patients hospitalized with community-acquired pneumonia. METHODS: A retrospective cohort study conducted at two tertiary teaching hospitals. Eligible subjects were admitted with a diagnosis of, had a chest x-ray consistent with, and had a discharge ICD-9 diagnosis of pneumonia. Subjects were excluded if they were "comfort measures only" or transferred from another acute care hospital. Subjects were considered to be on a medication if they were taking it at the time of presentation. RESULTS: Data was abstracted on 787 subjects at the two hospitals. Mortality was 9.2% at 30-days and 13.6% at 90-days. At presentation 52% of subjects were low risk, 34% were moderate risk, and 14% were high risk based on the pneumonia severity index. In the multivariable regression analysis, after adjusting for potential confounders including a propensity score, the use of statins at presentation (odds ratio 0.36, 95% confidence interval 0.14-0.92) was associated with decreased 30-day mortality. DISCUSSION: Prior outpatient statin use was associated with decreased mortality in patients hospitalized with community-acquired pneumonia despite their use being associated with comorbid illnesses likely to contribute to increased mortality. Confirmatory studies are needed, as well as research to determine the mechanism(s) of this protective effect. PMID- 16042798 TI - Risk of high blood pressure in salt workers working near salt milling plants: a cross-sectional and interventional study. AB - BACKGROUND: Workers working close to salt milling plants may inhale salt particles floating in the air, leading to a rise in plasma sodium, which, in turn, may increase the blood pressure and the risk of hypertension. METHODS: To test the above hypothesis, occupational health check-up camps were organized near salt manufacturing units and all workers were invited for a free health examination. The workers who worked with dry salt in the vicinity of salt milling plants were defined as "non-brine workers," while those working in brine pans located far away from milling plants were defined as "brine workers." Blood pressure (BP) was measured during each clinical examination. In all, 474 non brine workers and 284 brine workers were studied. RESULTS: Mean systolic blood pressure of non-brine workers (122.1 +/- 13.3 mm Hg) was significantly higher than that of brine workers (118.8 +/- 12.8 mm Hg, p < 0.01). Mean diastolic blood pressure of non-brine workers (71.5 +/- 10.4 mm Hg) was significantly higher than that of brine workers (69.7 +/- 9.4 mm Hg, p = 0.02). The prevalence of hypertension was significantly higher in non-brine workers (12.2%) than in brine workers (7.0%, p = 0.02). Nineteen salt workers were monitored while they used face masks and spectacles, for six days. Systolic, as well as diastolic, blood pressure of these workers began declining on the third day and continued to decline on the fourth day, but remained stationary up to the sixth day. The concentration of salt particles in the breathing zone of these workers was 376 mg/m3 air. CONCLUSION: Inhalation of salt particles in non-brine workers may be an occupational cause of increased blood pressure. PMID- 16042799 TI - CGH-Profiler: data mining based on genomic aberration profiles. AB - BACKGROUND: CGH-Profiler is a program that supports the analysis of genomic aberrations measured by Comparative Genomic Hybridisation (CGH). Comparative genomic hybridisation (CGH) is a well-established, molecular cytogenetic method that allows the detection of chromosomal imbalances in entire genomes. This technique is widely used in routine molecular diagnostics. Typically, chromosomal imbalances are described in a complex syntax based on the International Standard for Cytogenetic Nomenclature (ISCN). This semantic description of chromosomal imbalances hinders a large-scale statistical analysis across different experiments, e.g. for finding aberration patterns associated with a particular disease type or state. RESULTS: CGH-Profiler circumvents the semantic ISCN description by importing data from different CGH system vendors and by directly transferring the data into a table format that is readily accessible for subsequent statistical analysis. CGH-profiler comes with different consistency checks, calculates various statistics and automatically assigns a median copy number ratio to each chromosomal band. Import of CGH profiles from different CGH system vendors is already supported; its extension to other systems can be readily achieved through Perl scripts.CGH profiler can also be used to analyse comparative expressed sequence hybridisation (CESH) data. CESH reveals gene expression patterns according to chromosomal locations in a similar manner as CGH detects chromosomal imbalances. CONCLUSION: CGH-Profiler is a useful tool for processing of CGH and CESH data. PMID- 16042800 TI - GObar: a gene ontology based analysis and visualization tool for gene sets. AB - BACKGROUND: Microarray experiments, as well as other genomic analyses, often result in large gene sets containing up to several hundred genes. The biological significance of such sets of genes is, usually, not readily apparent. Identification of the functions of the genes in the set can help highlight features of interest. The Gene Ontology Consortium 1 has annotated genes in several model organisms using a controlled vocabulary of terms and placed the terms on a Gene Ontology (GO), which comprises three disjoint hierarchies for Molecular functions, Biological processes and Cellular locations. The annotations can be used to identify functions that are enriched in the set, but this analysis can be misleading since the underlying distribution of genes among various functions is not uniform. For example, a large number of genes in a set might be kinases just because the genome contains many kinases. RESULTS: We use the Gene Ontology hierarchy and the annotations to pick significant functions and pathways by comparing the distribution of functions in a given gene list against the distribution of all the genes in the genome, using the hypergeometric distribution to assign probabilities. GObar is a web-based visualizer that implements this algorithm. The public website for GObar 2 can analyse gene lists from the yeast (S. cervisiae), fly (D. Melanogaster), mouse (M. musculus) and human (H. sapiens) genomes. It also allows visualization of the GO tree, as well as placement of a single gene on the GO hierarchy. We analyse a gene list from a genomic study of pre-mRNA splicing to demonstrate the utility of GObar. CONCLUSION: GObar is freely available as a web-based tool at http://katahdin.cshl.org:9331/GO2 and can help analyze and visualize gene lists from genomic analyses. PMID- 16042801 TI - Socioeconomic factors and adolescent pregnancy outcomes: distinctions between neonatal and post-neonatal deaths? AB - BACKGROUND: Young maternal age has long been associated with higher infant mortality rates, but the role of socioeconomic factors in this association has been controversial. We sought to investigate the relationships between infant mortality (distinguishing neonatal from post-neonatal deaths), socioeconomic status and maternal age in a large, retrospective cohort study. METHODS: We conducted a population-based cohort study using linked birth-death certificate data for Missouri residents during 1997-1999. Infant mortality rates for all singleton births to adolescent women (12-17 years, n = 10,131; 18-19 years, n = 18,954) were compared to those for older women (20-35 years, n = 28,899). Logistic regression was used to estimate adjusted odds ratios (OR) and 95% confidence intervals (CI) for all potential associations. RESULTS: The risk of infant (OR 1.95, CI 1.54-2.48), neonatal (1.69, 1.24-2.31) and post-neonatal mortality (2.47, 1.70-3.59) were significantly higher for younger adolescent (12 17 years) than older (20-34 years) mothers. After adjusting for race, marital status, age-appropriate education level, parity, smoking status, prenatal care utilization, and poverty status (indicated by participation in WIC, food stamps or Medicaid), the risk of post-neonatal mortality (1.73, 1.14-2.64) but not neonatal mortality (1.43, 0.98-2.08) remained significant for younger adolescent mothers. There were no differences in neonatal or post-neonatal mortality risks for older adolescent (18-19 years) mothers. CONCLUSION: Socioeconomic factors may largely explain the increased neonatal mortality risk among younger adolescent mothers but not the increase in post-neonatal mortality risk. PMID- 16042802 TI - Computational tradeoffs in multiplex PCR assay design for SNP genotyping. AB - BACKGROUND: Multiplex PCR is a key technology for detecting infectious microorganisms, whole-genome sequencing, forensic analysis, and for enabling flexible yet low-cost genotyping. However, the design of a multiplex PCR assays requires the consideration of multiple competing objectives and physical constraints, and extensive computational analysis must be performed in order to identify the possible formation of primer-dimers that can negatively impact product yield. RESULTS: This paper examines the computational design limits of multiplex PCR in the context of SNP genotyping and examines tradeoffs associated with several key design factors including multiplexing level (the number of primer pairs per tube), coverage (the % of SNP whose associated primers are actually assigned to one of several available tube), and tube-size uniformity. We also examine how design performance depends on the total number of available SNPs from which to choose, and primer stringency criterial. We show that finding high multiplexing/high-coverage designs is subject to a computational phase transition, becoming dramatically more difficult when the probability of primer pair interaction exceeds a critical threshold. The precise location of this critical transition point depends on the number of available SNPs and the level of multiplexing required. We also demonstrate how coverage performance is impacted by the number of available snps, primer selection criteria, and target multiplexing levels. CONCLUSION: The presence of a phase transition suggests limits to scaling Multiplex PCR performance for high-throughput genomics applications. Achieving broad SNP coverage rapidly transitions from being very easy to very hard as the target multiplexing level (# of primer pairs per tube) increases. The onset of a phase transition can be "delayed" by having a larger pool of SNPs, or loosening primer selection constraints so as to increase the number of candidate primer pairs per SNP, though the latter may produce other adverse effects. The resulting design performance tradeoffs define a benchmark that can serve as the basis for comparing competing multiplex PCR design optimization algorithms and can also provide general rules-of-thumb to experimentalists seeking to understand the performance limits of standard multiplex PCR. PMID- 16042803 TI - L-glutamine therapy reduces endothelial adhesion of sickle red blood cells to human umbilical vein endothelial cells. AB - BACKGROUND: We have previously demonstrated that therapy with orally administered L-glutamine improves nicotinamide adenosine dinucleotide (NAD) redox potential of sickle red blood cells (RBC). On further analysis of L-glutamine therapy for sickle cell anemia patients, the effect of L-glutamine on adhesion of sickle RBC to human umbilical vein endothelial cells (HUVEC) was examined. METHODS: The first part of the experiment was conducted with the blood samples of the 5 adult sickle cell anemia patients who had been on L-glutamine therapy for at least 4 weeks on a dosage of 30 grams per day compared to those of patient control group. In the second part of the experiment 6 patients with sickle cell anemia were studied longitudinally. Five of these patients were treated with oral L-glutamine 30 grams daily and one was observed without treatment as the control. t-test and paired t-test were used for determination of statistical significance in cross sectional and longitudinal studies respectively. RESULTS: In the first study, the mean adhesion to endothelial cells with the autologous plasma incubated cells were 0.97 +/- 0.45 for the treated group and 1.91 +/- 0.53 for the nontreated group (p < 0.02). Similarly with lipopolysaccharide (LPS) incubated cells the mean adhesion to endothelial cells were 1.39 +/- 0.33 for the treated group and 2.80 +/- 0.47 for the untreated group (p < 0.001). With the longitudinal experiment, mean decrease in the adhesion to endothelial cells was 1.13 +/- 0.21 (p < 0.001) for the 5 treated patients whereas the control patient had slight increase in the adhesion to endothelial cells. CONCLUSION: In these studies, oral L-glutamine administration consistently resulted in improvement of sickle RBC adhesion to HUVEC. These data suggest positive physiological effects of L glutamine in sickle cell disease. PMID- 16042804 TI - Cardiac conduction abnormalities and congenital immunodeficiency in a child with Kabuki syndrome: case report. AB - BACKGROUND: Since it's recognition in 1981, a more complete phenotype of Kabuki syndrome is becoming evident as additional cases are identified. Congenital heart defects and a number of visceral abnormalities have been added to the typical dysmorphic features originally described. CASE REPORT: In this report we describe the clinical course of a child diagnosed with Kabuki syndrome based on characteristic clinical, radiological and morphologic features who died of a cardiac arrhythmia at 11-months of age. This infant, however, had abnormal pulmonary architecture and alterations in his cardiac conduction system resulting in episodes of bradycardia and asystole. This child also had an immunological phenotype consistent with common variable immunodeficiency. His clinical course consisted of numerous hospitalizations for recurrent bacterial infections and congenital hypogammaglobulinemia characterized by low serum IgG and IgA but normal IgM levels, and decreased antibody levels to immunizations. T-, B- and NK lymphocyte subpopulations and T-cell function studies were normal. CONCLUSION: This child may represent a more severe phenotype of Kabuki syndrome. Recurrent infections in a child should prompt a thorough immunological evaluation. Additionally, electrophysiology testing may be indicated if cardiopulmonary events occur which are not explained by anatomic defects. PMID- 16042805 TI - Gait dynamics in mouse models of Parkinson's disease and Huntington's disease. AB - BACKGROUND: Gait is impaired in patients with Parkinson's disease (PD) and Huntington's disease (HD), but gait dynamics in mouse models of PD and HD have not been described. Here we quantified temporal and spatial indices of gait dynamics in a mouse model of PD and a mouse model of HD. METHODS: Gait indices were obtained in C57BL/6J mice treated with the dopaminergic neurotoxin 1-methyl 4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, 30 mg/kg/day for 3 days) for PD, the mitochondrial toxin 3-nitropropionic acid (3NP, 75 mg/kg cumulative dose) for HD, or saline. We applied ventral plane videography to generate digital paw prints from which indices of gait and gait variability were determined. Mice walked on a transparent treadmill belt at a speed of 34 cm/s after treatments. RESULTS: Stride length was significantly shorter in MPTP-treated mice (6.6 +/- 0.1 cm vs. 7.1 +/- 0.1 cm, P < 0.05) and stride frequency was significantly increased (5.4 +/- 0.1 Hz vs. 5.0 +/- 0.1 Hz, P < 0.05) after 3 administrations of MPTP, compared to saline-treated mice. The inability of some mice treated with 3NP to exhibit coordinated gait was due to hind limb failure while forelimb gait dynamics remained intact. Stride-to-stride variability was significantly increased in MPTP-treated and 3NP-treated mice compared to saline-treated mice. To determine if gait disturbances due to MPTP and 3NP, drugs affecting the basal ganglia, were comparable to gait disturbances associated with motor neuron diseases, we also studied gait dynamics in a mouse model of amyotrophic lateral sclerosis (ALS). Gait variability was not increased in the SOD1 G93A transgenic model of ALS compared to wild-type control mice. CONCLUSION: The distinct characteristics of gait and gait variability in the MPTP model of Parkinson's disease and the 3NP model of Huntington's disease may reflect impairment of specific neural pathways involved. PMID- 16042806 TI - Pilot survey of oral health-related quality of life: a cross-sectional study of adults in Benin City, Edo State, Nigeria. AB - BACKGROUND: Oral health studies conducted so far in Nigeria have documented prevalence and incidence of dental disease using traditional clinical measures. However none have investigated the use of an oral health-related quality of life (OHRQoL) instrument to document oral health outcomes. The aims of this study are: to describe how oral health affects and impacts quality of life (QoL) and to explore the association between these affects and the oral health care seeking behavior of adults in Benin City, Edo State, Nigeria. METHODS: A cross-sectional survey recruited 356 adults aged 18-64 years from two large hospital outpatient departments and from members of a university community. Closed-ended oral health questionnaire with "effect and impact" item-questions from OHQoL-UK instrument was administered by trained interviewers. Collected data included sociodemographic, dental visits, and effects and impact of oral health on QoL. Univariate and bivariable analyses were done and a chi-square test was used to test differences in proportions. Multivariable analyses using ANOVA examined the association between QoL factors and visits to a dentist. RESULTS: Complete data was available for 83% of the participants. About 62% of participants perceived their oral health as affecting their QoL. Overall, 82%, 63%, and 77% of participants perceived that oral health has an effect on their eating or enjoyment of food, sleep or ability to relax, and smiling or laughing, respectively. Some 46%, 36%, and 25% of participants reported that oral health impact their daily activities, social activities, and talking to people, respectively. Dental visits within the last year was significantly associated with eating, speech, and finance (P < 0.05). The summary score for the oral health effects on QoL ranged from 33 to 80 with a median value of 61 (95% CI: 60, 62) and interquartile range of 52-70. Multivariable modeling suggested a model containing only education (F = 6.5, pr>F = 0.0111). The mean of effects sum score for those with secondary/tertiary education levels (mean = 61.8; 95% CI: 60.6, 62.9) was significantly higher than those with less than secondary level of education (mean = 57.2; 95% CI: 57.2, 60.6). CONCLUSION: Most adults in the study reported that oral health affects their life quality, and have little/no impact on their quality of life. Dental visits within the last year were associated with eating, speech, and finance. PMID- 16042807 TI - RNA amplification for successful gene profiling analysis. AB - The study of clinical samples is often limited by the amount of material available to study. While proteins cannot be multiplied in their natural form, DNA and RNA can be amplified from small specimens and used for high-throughput analyses. Therefore, genetic studies offer the best opportunity to screen for novel insights of human pathology when little material is available. Precise estimates of DNA copy numbers in a given specimen are necessary. However, most studies investigate static variables such as the genetic background of patients or mutations within pathological specimens without a need to assess proportionality of expression among different genes throughout the genome. Comparative genomic hybridization of DNA samples represents a crude exception to this rule since genomic amplification or deletion is compared among different specimens directly. For gene expression analysis, however, it is critical to accurately estimate the proportional expression of distinct RNA transcripts since such proportions directly govern cell function by modulating protein expression. Furthermore, comparative estimates of relative RNA expression at different time points portray the response of cells to environmental stimuli, indirectly informing about broader biological events affecting a particular tissue in physiological or pathological conditions. This cognitive reaction of cells is similar to the detection of electroencephalographic patterns which inform about the status of the brain in response to external stimuli. As our need to understand human pathophysiology at the global level increases, the development and refinement of technologies for high fidelity messenger RNA amplification have become the focus of increasing interest during the past decade. The need to increase the abundance of RNA has been met not only for gene specific amplification, but, most importantly for global transcriptome wide, unbiased amplification. Now gene-specific, unbiased transcriptome wide amplification accurately maintains proportionality among all RNA species within a given specimen. This allows the utilization of clinical material obtained with minimally invasive methods such as fine needle aspirates (FNA) or cytological washings for high throughput functional genomics studies. This review provides a comprehensive and updated discussion of the literature in the subject and critically discusses the main approaches, the pitfalls and provides practical suggestions for successful unbiased amplification of the whole transcriptome in clinical samples. PMID- 16042808 TI - Adrenal metastasis as first presentation of hepatocellular carcinoma. AB - BACKGROUND: Metastases from hepatocellular carcinoma (HCC) can be found in the lung and adrenal gland. We report case of a patient who presented with adrenal metastasis as the first clinical manifestation of HCC. CASE PRESENTATION: A patient was referred for surgical treatment for a tumor in retro-peritoneal space. The computerized tomography (CT) scan revealed a mass originating from the left adrenal gland. The patient underwent left adrenalectomy and the exploration of abdominal cavity did not reveal any other palpable lesions. Histologically, the resected lesion was a poorly differentiated metastatic tumor from HCC. Seven months later patient was readmitted complaining of cachexia, icterus, and significant weight loss. CT scan revealed hyperdense lesions of the liver. CONCLUSION: HCC may have atypical presentations like in present case. Fine needle aspiration/tru-cut biopsy might be useful in the investigation of an accidentally discovered adrenal mass regardless of the size and can lead to the detection of a primary tumor. PMID- 16042809 TI - Total blood lymphocyte counts in hemochromatosis probands with HFE C282Y homozygosity: relationship to severity of iron overload and HLA-A and -B alleles and haplotypes. AB - BACKGROUND: It has been reported that some persons with hemochromatosis have low total blood lymphocyte counts, but the reason for this is unknown. METHODS: We measured total blood lymphocyte counts using an automated blood cell counter in 146 hemochromatosis probands (88 men, 58 women) with HFE C282Y homozygosity who were diagnosed in medical care. Univariate and multivariate analyses of total blood lymphocyte counts were evaluated using these variables: sex; age, transferrin saturation, and serum ferritin concentration at diagnosis; units of blood removed by phlebotomy to achieve iron depletion; and human leukocyte antigen (HLA)-A and -B alleles and haplotypes. RESULTS: The mean age at diagnosis was 49 +/- 14 years (range 18 - 80 years) in men and 50 +/- 13 years (range 22-88 years) in women. The correlations of total blood lymphocyte counts with sex, age, transferrin saturation, and serum ferritin concentration at diagnosis, and units of blood removed by phlebotomy to achieve iron depletion were not significant at the 0.05 level. Univariate analyses revealed significant associations between total blood lymphocyte counts and presence of the HLA-A*01, -B*08, and -B*14 alleles, and the A*01-B*08 haplotype. Presence of the A*01 allele, B*08 allele, or A*01-B*08 haplotype were associated with a lower total blood lymphocyte count, whereas presence of the B*14 allele was associated with a greater total blood lymphocyte count. There was an inverse association of total blood lymphocyte count with units of phlebotomy to achieve iron depletion, serum ferritin concentration, and with presence of the A*01-B*08 haplotype. CONCLUSION: We conclude that there is a significant inverse relationship of total blood lymphocyte counts and severity of iron overload in hemochromatosis probands with HFE C282Y homozygosity. The presence of the HLA-A*01 allele or the -B*08 allele was also associated with significantly lower total blood lymphocyte counts, whereas presence of the -B*14 allele was associated with significantly higher total blood lymphocyte counts. In univariate and multivariate analyses, total blood lymphocyte counts were significantly lower in probands with the HLA-A*01 B*08 haplotype than in probands without this haplotype. PMID- 16042810 TI - Intracardiac thrombus in Behcet's disease: two case reports. AB - Intracardiac thrombus in Behcet's disease is an extremely rare manifestation. We report two such cases. A 20-year-old man presented with dyspnoea, cough and haemoptysis. Right heart thrombus associated with pulmonary artery aneurysm and thromboembolism was identified by helical CT and transoesophageal echocardiography. The second case was a 29-year-old male admitted for fever and chest pain. A diagnosis of right atrial thrombosis associated with pulmonary embolism and hyperhomocysteinemia was made. Due to the absence of haemodynamic compromise, medical management consisting of immunosupressive and anticoagulation therapy was adopted which resulted in complete dissolution of the thrombus with dramatic clinical improvement in both cases of clinical status. CONCLUSION: intracardiac thrombus is a rare complication of Behcet's disease. As shown in our patients, medical treatment should be considered as the first line. PMID- 16042811 TI - Conjugative plasmid pAW63 brings new insights into the genesis of the Bacillus anthracis virulence plasmid pXO2 and of the Bacillus thuringiensis plasmid pBT9727. AB - BACKGROUND: Bacillus cereus, Bacillus anthracis and Bacillus thuringiensis belong to the genetically close-knit Bacillus cereus sensu lato group, a family of rod shaped Gram-positive bacteria. pAW63 is the first conjugative plasmid from the B. cereus group to be completely sequenced. RESULTS: The 71,777 bp nucleotide sequence of pAW63 reveals a modular structure, including a 42 kb tra region encoding homologs of the Type IV secretion systems components VirB11, VirB4 and VirD4, as well as homologs of Gram-positive conjugation genes from Enterococcus, Lactococcus, Listeria, Streptococcus and Staphylococcus species. It also firmly establishes the existence of a common backbone between pAW63, pXO2 from Bacillus anthracis and pBT9727 from the pathogenic Bacillus thuringiensis serovar konkukian strain 97-27. The alignment of these three plasmids highlights the presence of well conserved segments, in contrast to distinct regions of high sequence plasticity. The study of their specific differences has provided a three point reference framework that can be exploited to formulate solid hypotheses concerning the functionalities and the molecular evolution of these three closely related plasmids. This has provided insight into the chronology of their divergence, and led to the discovery of two Type II introns on pAW63, matching copies of the mobile element IS231L in different loci of pXO2 and pBT9727, and the identification on pXO2 of a 37 kb pathogenicity island (PAI) containing the anthrax capsule genes. CONCLUSION: The complete sequence determination of pAW63 has led to a functional map of the plasmid yielding insights into its conjugative apparatus, which includes T4SS-like components, as well as its resemblance to other large plasmids of Gram-positive bacteria. Of particular interest is the extensive homology shared between pAW63 and pXO2, the second virulence plasmid of B. anthracis, as well as pBT9727 from the pathogenic strain B. thuringiensis serovar konkukian strain 97-27. PMID- 16042812 TI - Too much or too little step width variability is associated with a fall history in older persons who walk at or near normal gait speed. AB - BACKGROUND: Decreased gait speed and increased stride time, stride length, double support time, and stance time variability have consistently been associated with falling whereas step width variability has not been strongly related to falls. The purpose was to examine the linear and nonlinear associations between gait variability and fall history in older persons and to examine the influence of gait speed. METHODS: Gait characteristics and fall history were obtained in 503 older adults (mean age = 79; 61% female) participating in the Cardiovascular Health Study who could ambulate independently. Gait characteristics were recorded from two trials on a 4 meter computerized walkway at the subject's self-selected walking speed. Gait variability was calculated as the coefficient of variation. The presence of a fall in the past 12 months was determined by interview. The nonlinear association between gait variability and fall history was examined using a simple three level classification derived from the distribution of the data and from literature based cut-points. Multivariate logistic regression was used to examine the association between step width variability (extreme or moderate) and fall history stratifying by gait speed (1.0 m/s) and controlling for age and gender. RESULTS: Step length, stance time, and step time variability did not differ with respect to fall history (p > .33). Individuals with extreme step width variability (either low or high step width variability) were more likely to report a fall in the past year than individuals with moderate step width variability. In individuals who walked > or = 1.0 m/s (n = 281), after controlling for age, gender, and gait speed, compared to individuals with moderate step width variability individuals with either low or high step width variability were more likely to have fallen in the past year (OR and 95% CI 4.38 [1.79-10.72]). The association between step width variability and fall history was not significant in individuals who walked < 1.0 m/s (n = 224). CONCLUSION: Extreme (either too little or too much) step width variability is associated with falls in the past year in older persons who walk at or near normal gait speed and not in older persons who walk slowly (< 1.0 m/s). PMID- 16042813 TI - Quinolinic acid selectively induces apoptosis of human astrocytes: potential role in AIDS dementia complex. AB - There is evidence that the kynurenine pathway (KP) and particularly one of its end products, quinolinic acid (QUIN) play a role in the pathogenesis of several major neuroinflammatory diseases, and more particularly AIDS dementia complex (ADC). We hypothesized that QUIN may be involved in astrocyte apoptosis because: 1) apoptotic astrocytes have been observed in the brains of ADC patients, 2) ADC patients have elevated cerebrospinal fluid QUIN concentrations, and 3) QUIN can induce astrocyte death. Primary cultures of human fetal astrocytes were treated with three pathophysiological concentrations of QUIN. Numeration of apoptotic cells was assessed using double immunocytochemistry for expression of active caspase 3 and for nucleus condensation. We found that treatment of human astrocytes with QUIN induced morphological (cell body shrinking) and biochemical changes (nucleus condensation and over-expression of active caspase 3) of apoptosis. After 24 hours of treatment with QUIN 500 nM and 1200 nM respectively 10 and 14% of astrocytes were undergoing apoptosis. This would be expected to lead to a relative lack of trophic support factors with consequent neuronal dysfunction and possibly death. Astroglial apoptosis induced by QUIN provides another potential mechanism for the neurotoxicity of QUIN during ADC. PMID- 16042814 TI - Information theory-based algorithm for in silico prediction of PCR products with whole genomic sequences as templates. AB - BACKGROUND: A new algorithm for assessing similarity between primer and template has been developed based on the hypothesis that annealing of primer to template is an information transfer process. RESULTS: Primer sequence is converted to a vector of the full potential hydrogen numbers (3 for G or C, 2 for A or T), while template sequence is converted to a vector of the actual hydrogen bond numbers formed after primer annealing. The former is considered as source information and the latter destination information. An information coefficient is calculated as a measure for fidelity of this information transfer process and thus a measure of similarity between primer and potential annealing site on template. CONCLUSION: Successful prediction of PCR products from whole genomic sequences with a computer program based on the algorithm demonstrated the potential of this new algorithm in areas like in silico PCR and gene finding. PMID- 16042815 TI - Brands, costs and registration status of antimalarial drugs in the Kenyan retail sector. AB - BACKGROUND: Although an important source of treatment for fevers, little is known about the structure of the retail sector in Africa with regard to antimalarial drugs. This study aimed to assess the range, costs, sources and registration of antimalarial drugs in the Kenyan retail sector. METHODS: In 2002, antimalarial drug registration and trade prices were established by triangulating national registration lists, government gazettes and trade price indices. Data on registration status and trade prices were compared with similar data generated through a retail audit undertaken among 880 randomly sampled retailers in four districts of Kenya. RESULTS: Two hundred and eighteen antimalarial drugs were in circulation in Kenya in 2002. These included 65 "sulfur"-pyrimethamine (sulfadoxine-pyrimethamine and sulfalene-pyrimethamine (SP), the first-line recommended drug in 2002) and 33 amodiaquine (AQ, the second-line recommended drug) preparations. Only half of SP and AQ products were registered with the Pharmacy and Poisons Board. Of SP and AQ brands at district level, 40% and 44% were officially within legal registration requirements. 29% of retailers at district level stocked SP and 95% stocked AQ. The retail price of adult doses of SP and AQ were on average 0.38 and 0.76 US dollars, 100% and 347% higher than trade prices from manufacturers and importers. Artemether-lumefantrine, the newly announced first-line recommended antimalarial drug in 2004, was found in less than 1% of all retail outlets at a median cost of 7.6 US dollars. CONCLUSION: There is a need to ensure that all antimalarial drugs are registered with the Pharmacy and Poisons Board to facilitate a more stringent post-marketing surveillance system to ensure drugs are safe and of good quality post registration. PMID- 16042863 TI - Total knee replacement for posttraumatic degenerative arthritis of the knee. AB - OBJECTIVE: To evaluate the results of total knee arthroplasty (TKA) in patients with posttraumatic degenerative arthritis due to a previous fracture around the knee. METHODS: We analyzed the results of 15 TKAs, performed from 1997 to 2003, in 15 patients with post-traumatic degenerative arthritis due to a previous fracture around knee. There were 3 women and 12 men with an average age of 58 years (range, 31-76 years). The time from fracture to arthroplasty averaged 8.2 years (range, 2-27 years). Internal fixation had previously been performed in 8 patients resulting in retained hardware. At the time of arthroplasty a femoral fracture malunion was present in two knees. Lateral retinacular release (4 knees), extensor mechanism realignment (1 knee) or medial collateral ligament reconstruction (1 knee) were needed at the time of arthroplasty. RESULTS: Follow up averaged 35 months (range, 12-73 months). No patient was lost for follow-up. According to the Knee Society Score scale, the mean preoperative knee score was 37 (range, 10-70) and functional score was 41 (range, 0-60). They were improved significantly to a mean of 84 (range, 10-100) and 76 (range, 20-100) points, respectively at the latest follow-up. The mean knee arc of motion were improved from 84 degree preoperation to 94 degree at the latest follow-up. Postoperative manipulation under anesthesia for poor motion was carried out in 4 knees. No knee had aseptic loosening that required subsequent revision. Two knees developed superficial infection and were treated with debridement. It subsequently recovered with the retention of components. CONCLUSIONS: Significant improvement in function and relief of pain has been achieved in patients with previous fractures undergoing subsequent TKA. However, this procedure is technically demanding and patients are at increased risk for restricted motion and need more care following TKA. This study suggests that the outcome of TKA may be improved further by making special efforts to restore limb alignment, to ensure correct component positioning, and to manage soft tissue balance. PMID- 16042816 TI - The influence of methylphenidate on the power spectrum of ADHD children - an MEG study. AB - BACKGROUND: The present study was dedicated to investigate the influence of Methylphenidate (MPH) on cortical processing of children who were diagnosed with different subtypes of Attention Deficit Hyperactivity Disorder (ADHD). As all of the previous studies investigating power differences in different frequency bands have been using EEG, mostly with a relatively small number of electrodes our aim was to obtain new aspects using high density magnetoencephalography (MEG). METHODS: 35 children (6 female, 29 male) participated in this study. Mean age was 11.7 years (+/- 1.92 years). 17 children were diagnosed of having an Attention Deficit/Hyperactivity Disorder of the combined type (ADHDcom, DSM IV code 314.01); the other 18 were diagnosed for ADHD of the predominantly inattentive type (ADHDin, DSM IV code 314.0). We measured the MEG during a 5 minute resting period with a 148-channel magnetometer system (MAGNES 2500 WH, 4D Neuroimaging, San Diego, USA). Power values were averaged for 5 bands: Delta (D, 1.5-3.5 Hz), Theta (T, 3.5-7.5 Hz), Alpha (A, 7.5-12.5 Hz), Beta (B, 12.5-25 Hz) and Global (GL, 1.5-25 Hz).). Additionally, attention was measured behaviourally using the D2 test of attention with and without medication. RESULTS: The global power of the frequency band from 1.5 to 25 Hz increased with MPH. Relative Theta was found to be higher in the left hemisphere after administration of MPH than before. A positive correlation was found between D2 test improvement and MPH-induced power changes in the Theta band over the left frontal region. A linear regression was computed and confirmed that the larger the improvement in D2 test performance, the larger the increase in Theta after MPH application. CONCLUSION: Main effects induced by medication were found in frontal regions. Theta band activity increased over the left hemisphere after MPH application. This finding contradicts EEG results of several groups who found lower levels of Theta power after MPH application. As relative Theta correlates with D2 test improvement we conclude that MEG provide complementary and therefore important new insights to ADHD. PMID- 16042864 TI - Effect of Shenfu injection on nuclear factor-kappaB during myocardial ischemia/reperfusion injury in rats. AB - OBJECTIVE: To investigate effects of Shenfu injection on the concentrations of plasma tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), activity of Nuclear Factor kappa B (NF-kappaB) and heart tissue ultrastructure during myocardial ischemia/reperfusion (I/R) injury in rats and its potential mechanism. METHODS: Myocardial ischemia/reperfusion (I/R) was produced by ligation and release of the left anterior descending coronary artery. Ischemia lasted for 30 min and reperfusion for 60 min. Twenty-four healthy male SD rats weighing 230-280 g were randomly divided into three groups (n = 8, each): Group I (Sham-operation group); Group II (I/R group); Group III (Shenfu group), in which Shenfu injection (10 ml/kg) was intraperitoneally injected 30 min before ischemia in animals with I/R. The plasma concentrations of IL-6 and TNF-alpha were measured by ELISA, and the heart was harvested for determination of NF-kappaB levels by Ecl-western blot analysis. Electron microscopy was used to study its ultrastructure. RESULTS: After reperfusion, NF-kappaB binding activity in myocardial nuclei and the plasma concentrations of IL-6 and TNF-alpha were significantly increased in Group II, compared with Group I (P < 0.01), and they were markedly reduced in Group III, compared with Group II (P < 0.01). In addition, electron microscopic examination showed more serious injury of the myocardium ultrastructure in Group II, while in Group III the myocardial ultrastructure was similar to normal state. CONCLUSIONS: Shenfu injection inhibits NF-kappaB activity in I/R myocardium and leads to down regulation of proinflammatory cytokine expression, which might be one of the molecular mechanisms of Shenfu injection in cardioprotection. PMID- 16042865 TI - Analysis of closed abdominal injury in pregnant women. AB - OBJECTIVE: To explore the characteristics of closed abdominal injury in pregnancy women and its treatment. METHODS: The clinical data of 37 pregnancy patients with closed abdominal injury treated in our hospital from June 1993 to June 2003 were collected and analyzed. RESULTS: All the 37 patients were treated with operation. Among them 2 early pregnancy patients with intestinal rupture and 1 patient with retroperitoneal hematoma were treated under laparoscope; in other 34 pregnancy patients laparotomy was performed. Of the 34 patients 8 used cesarean section because premature separation of placenta and enlarged womb interrupted the management of intra-abdominal organ injury. In the 37 patients 33 (89.1%) were cured, 4 (10.8%) die, postoperative complication rate was 16.2% (6/37). Two patients (5.4%) suffered from abdominal cavity infection, 3 (8.1%) from pulmonary infection, and 1 (2.7%) had multi-organ failure. CONCLUSIONS: For pregnancy patients with closed abdominal injury, besides obsteric diseases intra-abdominal injury should be given much attention. Accurate diagnosis and timely treatment can gain the time to save the life of both mother and fetus. PMID- 16042866 TI - Effect of nitric oxide-induced tyrosine phosphorylation of calcium-activated potassium channel alpha subunit on vascular hyporesponsiveness in rats. AB - OBJECTIVE: To study the effect of nitric oxide-induced tyrosine phosphorylation of large-conductance calcium-activated potassium (BK(Ca)) channel alpha subunit on vascular hyporesponsiveness in rats. METHODS: A total of 46 Wistar rats of either sex, weighing 250 g +/- 20 g, were used in this study. Models of vascular hyporesponsiveness induced by hemorrhagic shock (30 mm Hg for 2 hours) in vivo and by L-arginine in vitro were established respectively. The vascular responsiveness of isolated superior mesenteric arteries to norepinephrine was observed. Tyrosine phosphorylation of BK(Ca) alpha subunit was evaluated with methods of immunoprecipitation and Western blotting. RESULTS: In the smooth muscle cells of the superior mesenteric arteries, the expression of BK(Ca) alpha subunit tyrosine phosphorylation increased following hemorrhagic shock, and L arginine could induce BK(Ca )channel alpha subunit tyrosine phosphorylation in a time- and dose-dependent manner. L-NAME (Nomega-nitro-L-arginine-methyl-ester), a nitric oxide synthetase inhibitor, could partly restore the decreased vasoresponsiveness of the superior mesenteric arteries after hemorrhagic shock in rats. Down-regulating the protein tyrosine phosphorylation with genistein, a widely-used special protein tyrosine kinase inhibitor, could partly improve the decreased vasoresponsiveness of the superior mesenteric arteries induced by L arginine in vitro, while up-regulating the protein tyrosine phosphorylation with Na(3)VO(4), a protein tyrosine phosphatase inhibitor, could further decrease the nitric oxide-induced vascular hyporesponsiveness, which could be partly ameliorated by 0.1 mmol/L tetrabutylammonium chloride (TEA), a selective BK(Ca )inhibitor at this concentration. CONCLUSIONS: Nitric oxide can induce the tyrosine phosphorylation of BK(Ca) alpha subunit, which influences the vascular hyporesponsiveness in hemorrhagic shock rats or induced by L-arginine in vitro. PMID- 16042867 TI - Monocyte chemoattractant protein-1 level in serum of patients with acute spinal cord injury. AB - OBJECTIVE: To detect the concentration of monocyte chemoattractant protein-1 (MCP 1) in the serum of patients with incomplete spinal cord injury and evaluate its relation with the pathologic classification of the spinal cord injury. METHODS: MCP-1 concentration in the serum of patients with incomplete spinal cord injury (iSCI), single spine compression and healthy subjects were detected by ELISA, respectively in the present study and the magnetic resonance imaging data of these patients were studied at the same time on a blind base. RESULTS: Serum level of MCP-1 in iSCI patients was 428 pg/ml +/- 11 pg/ml by ELISA, which was higher than both that of the patients with single spine compression and of controls, with the concentration of 184 pg/ml +/- 21 pg/ml and 124 pg/ml +/- 15 pg/ml, respectively. There was significant difference between any two groups (P < 0.01). iSCI patients with normal MRI showed a lower serum level of MCP-1 as 312 pg/ml+/- 30 pg/ml. Pathological classification of spinal cord edema and hematoma corresponded to 390 pg/ml +/- 16 pg/ml and 508 pg/ml+/- 24 pg/ml in the concentration of MCP-1. CONCLUSIONS: MCP-1 may induce secondary inflammatory response by recruiting inflammatory cells to the injury site and thus affect the prognosis of spinal cord injury. PMID- 16042868 TI - Expression and effect of Caspase-3 in neurons after tractive spinal cord injury in rats. AB - OBJECTIVE: To investigate Caspase-3 expression and its role in neuronal apoptosis. METHODS: The T13-L2 spinal cord of rats was injured by traction after the amplitude of P1-N1 wave, monitored by a cortical somatosensory evoked potential (CSEP) monitor, decreased to seventy percent of that before operation. Then rats were killed in 6 h, 1 d, 4 d, 7 d, 14 d and 21 d respectively after operation. Flow cytometer terminal deoxynucleotldyl transferease-mediated biotinylated deoxynuridine triphosphate nick end labeling (TUNEL), Caspase-3 activity assay and immunohistochemical method were applied to investigate Caspase 3 expression in the spinal cord tissue and to study neuronal apoptosis in rats. RESULTS: After spinal cord injury, apoptotic cells detected by flow cytometry and TUNEL-positive cells were significantly more, and positive immunohistochemical staining of Caspase-3 and Caspase-3 activity were significantly higher in Group injury than in Groups control and laminectomy, respectively (P > 0.05, P > 0.01). Similar trend of changes was noticed in apoptotic cells, TUNEL-positive cells and positive immunohistochemical staining of Caspase-3, all of which reached their respective peak 7 days after operation. Caspase-3 activity reached its peak, however, 4 days postoperatively. CONCLUSIONS: Increased expression and activity of Caspase-3 protein in neurons after tractive spinal cord injury is the biochemical signal of early spinal cell apoptosis. It is of great significance for understanding the mechanism of spinal cord injury. PMID- 16042869 TI - Pyrroloquinoline quinone enhances regeneration of transected sciatic nerve in rats. AB - OBJECTIVE: To investigate the effect of pyrroloquinoline quinone (PQQ) on nerve regeneration of transected sciatic nerve in animal models. METHODS: Forty SD rats weighing 220-240 g were randomized into a PQQ group (n = 20) and a control group (n = 20). Each animal underwent sciatic nerve transection operation. After the operation, PQQ 0.5 ml (250 microg/Kg) was injected at the operation site in the PQQ group, while the same volume of normal saline was delivered in the control group. Nerve functional evaluation, electrophysiological index recording were carried out according to the experimental design. Newly generated nerve specimens were harvested 12 weeks postoperatively for morphological studies. RESULTS: In the PQQ group there was a good nerve regeneration and the sciatic nerve function, sciatic nerve function index, electrophysiological index and morphological appearance were superior to the control group (P < 0.05). CONCLUSIONS: PQQ has a remarkable effect in enhancing nerve regeneration of transected peripheral nerve. PMID- 16042870 TI - Unilateral external fixator in the treatment of lower third humeral shaft fractures. AB - OBJECTIVE: To retrospectively analyze the effect of unilateral external fixators in the treatment of lower third humeral shaft fractures. METHODS: From October 1997 to October 2003, 33 patients aged 15 -70 years (average 31 years) with lower third humeral shaft fractures were treated with unilateral external fixators. There were 9 spiral fractures (type A1), 1 oblique fracture (type A2), 3 transverses fractures (type A3) and 20 comminuted fractures (11 type B1, 9 type B2) according to AO classification. Fifteen cases were treated with open reduction and limited internal fixation and fixation with external fixators, 10 cases treated with open reduction and fixation with external fixators, and 8 cases treated with closed reduction and fixation with external fixators. Nerve exploration was undertaken in 9 cases with preoperative radial nerve injury. External fixators were removed after bone healing. The average follow-up was 18 months with a range from 8 to 24 months. RESULTS: The time of bone healing ranged 11-22 weeks (average 14 weeks). The latest follow-up showed the functions of 9 cases of preoperative radial nerve injury and of 2 cases postoperative radial nerve injury and the function of elbow were recovered to normal. There were only 7 cases of superficial infection at pin hole, which was subsided by using oral antibiotics and pin-hole care with mild disinfectants. CONCLUSIONS: Fixation with unilateral external fixators combined with open reduction and limited internal fixation has a good effect in the treatment of lower third humeral shaft fractures. PMID- 16042871 TI - Application of ulinastatin in severe craniocerebral injuries. AB - OBJECTIVE: To assess the therapeutic effect of ulinastatin on severe craniocerebral injuries and to explore its mechanism. METHODS: There were 87 cases of severe brain injury in this series and they were either treated by ulinastatin (treatment group, 41 cases) or not (control group, 46 cases) besides routine managements. We estimated C-reactive protein, interleukin-6, superoxide dismutase, and endothelin from plasmas of all the cases on the 1st, 3rd, 5th, and 7th day after injury. RESULTS: C-reactive protein level rose on the 1st and 3rd day after injury in the two groups, but descended in treatment group on the 5th and 7th day and was significantly lower than that in control group (P < 0.01). No significant difference was found for interleukin-6 in two groups during 1-5 days after injury, but on the 7th day, it decreased significantly in treatment group than control one (P < 0.01). Superoxide dismutase was higher in treatment group than control one in 5-7 days after injury (P < 0.01). Endothelin elevated on the 1st day after injury but dropped afterwards in the two groups, in which the level in treatment group was lower than that in control one. The incidence of gastrointestinal hemorrhage was lower in treatment group than control one (P < 0.01). CONCLUSIONS: Ulinastatin has the function of protecting cerebral tissue, reducing the incidence of gastrointestinal hemorrhage, improving hepatic and renal function and prognosis. PMID- 16042872 TI - Octyl-a-cyanoacrylate adhesive in the treatment of tibial transverse fracture in rabbits. AB - OBJECTIVE: To observe the effect of octyl-a-cyanoacrylate upon bone healing and its degradation in vitro after middle tibial transverse fracture in rabbitsi and to establish treatment of higher efficacy with the application of octyl-a cyanoacrylate. METHODS: Middle tibial transverse fracture model of New Zealand rabbits was established. In the experimental group, internal fixation with 2 mm Kirschner wires was performed and the broken ends were fixed with octyl-a cyanoacrylate. In the control group, only internal fixation with 2 mm Kirschner wires was conducted. Animals were killed at preset time intervals of 2, 4, 6, 8, 10 and 12 weeks postoperatively and samples were harvested. RESULTS: Two weeks after operation, clear fracture lines were observed in both the experimental and the control groups. Fibrous soft tissue connection was noted between the broken ends and there was soft tissue adhesion around the fracture site. There was no callus formation and the broken ends were surrounded by adhesive soft tissues. Obvious external callus formation was confirmed at 8 weeks after operation in both groups with partial disappearance of fracture lines. Ten and twelve weeks after the operation, fracture lines disappeared completely and there was obvious external callus formation and bone union. In the fourth week, fibrous cells and chondrocytes were found to grow into the colloid and surround it at the 6th week. The adhesive material was degraded and gradually absorbed at the 8th week. Chondrification was observed. CONCLUSIONS: Two weeks after fixation for tibial fracture in rabbits, octyl-a-cyanoacrylate begins in vivo degradation. Chondrocytes and fibrocytes gradually grow into the degradation area and surround the adhesive material, which broke into pieces at 8 weeks. Complete degradation and disappearance of the adhesive material is present between 10 and 12 weeks. No barrier effect hampering fracture healing is noted. PMID- 16042873 TI - Dynamic change of serum protein S100b and its clinical significance in patients with traumatic brain injury. AB - OBJECTIVE: To analyze the dynamic change of serum protein S100b in patients with traumatic brain injury and its clinical value in assessing brain damage. METHODS: According to Glasgow coma scale (GCS), 102 cases of traumatic brain injury were divided into mild brain injury group (GCS > or = 13, n = 31, Group A), moderate brain injury group (8 < GCS < 13, n = 37, Group B) and severe brain injury group (GCS < or = 8, n = 34, Group C). Serial S100b concentrations were analyzed by enzyme-linked immunosorbent assay (ELISA) in blood samples taken on admission, 12 h, 24 h, 48 h, 72 h and 7 days after traumatic brain injury. RESULTS: The severe brain injury group showed significantly higher concentration of serum S100b, with earlier increase and longer duration, than the mild and moderate brain injury groups. The patients with higher S100b exhibited lower GCS scores and poor clinical prognosis. The increase in S100b could emerge before clinical image evidence indicated so. CONCLUSIONS: Serum S100b can be used as a sensitive index for assessment and prediction of traumatic brain injury severity and prognosis. PMID- 16042874 TI - Comparative study on cerebral injury after open heart surgery in patients with congenital and rheumatic heart disease. AB - OBJECTIVE: To comparatively study the different effects of open heart surgery on brain tissues of patients with congenital and rheumatic heart disease. METHODS: Forty patients with congenital heart disease (CHD, CHD group, n = 20) or rheumatic heart disease (RHD, RHD group, n = 20) underwent on-pump (cardiopulmonary bypass, CPB) heart-beating open heart surgery. Blood samples before CPB, and 20 minutes, 1 hour, 24 hours and 7 days after CPB were collected, and the levels of neuron-specific enolase (NSE) and protein S-100b in the plasma were determined with enzyme-linked immunoadsorbent assay (ELISA), respectively. All the patients were examined with electroencephalogram (EEG) before and 1 week after operation. The changes of NSE, S-100b and EEG compared to verify the difference of postoperative cerebral injury between CHD cases and RHD cases. RESULTS: The plasma level of S-100b increased significantly 20 minutes after CPB and was still higher than the preoperative level at 24 hours after operation in both groups (P < 0.01). The plasma level of NSE increased more significantly in the CHD group than in the RHD group 20 minutes after CPB and it returned to the normal level 24 hours after CPB in the CHD group but remained at a high level in the RHD group (P < 0.01). The levels of NSE and S-100b returned to the normal levels on the 7th day after CPB. Abnormal EEG was found in 75% of the patients in the CHD group and 60% in the RHD group. CONCLUSIONS: On-pump heart-beating open heart surgery can cause certain cerebral injury in the patients with CHD or RHD. The injury was more severe and recovered more quickly in the CHD group than in the RHD group. PMID- 16042875 TI - Double electrodes simultaneous stimulation and implantation technique in deep brain stimulation. AB - Posttraumatic tremor is often one of the causes of disability in head injury patients. Usually, pharmacotherapy for this type of tremor is not effective. Since early 1970s, surgical ablation of the ventral thalamus has been used to treat various types of tremor. Nowadays, deep brain stimulation (DBS) confirms its efficacy in alleviating different forms of tremor, including posttraumatic tremor. Such therapy has been reported achieving around 80% success rate in the treatment of posttraumatic tremor. These successful results suggest that the application of DBS therapy can be considered as one of the alternative treatments for minimizing the tremor occurring from different pathologies. PMID- 16042876 TI - [Challenges regarding the treatment of viral hepatitis B]. PMID- 16042877 TI - [Molecular cloning and identification of a novel gene and its relationship to hepatocellular carcinoma]. PMID- 16042878 TI - [A study of the dosage and efficacy of entecavir for treating hepatitis B virus]. AB - OBJECTIVE: To evaluate the antiviral activity and safety of entecavir in patients with chronic HBV infection as a preliminarily step in selecting 0.1 mg or 0.5 mg as a better dosage for a further large scale clinical trial. METHODS: This was a randomized, double-blinded, placebo-controlled and dose-ranging trial of entecavir usage in 212 patients with chronic HBV infection. The patients were randomly assigned to 3 groups: 0.1 mg entecavir (69), 0.5 mg entecavir (72) and, placebo (71) groups and treated for 28 days. The patients were then followed for 56 days without treatment. RESULTS: The proportion of subjects who achieved the primary endpoint at day 28, with their HBV DNA level decreased >2 log or undetectable, was significantly greater in the entecavir 0.1 mg and 0.5 mg dose groups compared with the placebo group (P < 0.01 for both comparisons). The mean change from baseline in HBV DNA levels at day 28 was greater for entecavir 0.1mg and 0.5 mg groups compared with the placebo group (both P < 0.01). The mean change from baseline in HBV DNA levels at day 28 for entecavir 0.5 mg group was greater than that of the entecavir 0.1 mg group (P < 0.01). During the 56-day post-dosing follow-up phase, the entecavir 0.5 mg group was associated with greater and more sustained suppression of viral replication than the entecavir 0.1 mg group (P < 0.01). There were no clinically meaningful differences in the incidence of any adverse events between the entecavir dosing and the placebo groups. CONCLUSION: Entecavir at both 0.1 mg and 0.5 mg doses demonstrated superior antiviral activity compared with a placebo. Since the entecavir 0.5 mg dose appears to have greater antiviral activity than the 0.1 mg dose and with a comparable safety and tolerability profile, the 0.5 mg entecavir dose could be used in further trials. PMID- 16042879 TI - [Influences of the genotypes of HBV and HBeAg regarding their response to PEG-IFN in chronic hepatitis B patients]. AB - OBJECTIVE: To study the effects of genotypes of HBV and HBeAg on the response to PEG-interferon alpha (PEG-IFN) in chronic hepatitis B (CHB) patients. METHODS: PCR-RFLP and S gene sequencing were conducted in 42 CHB patients. RESULTS: The sustained response (SR) rates were 66.7% in genotype B and 27.3% in genotype C group. The P value was 0.039 by the Pearson Chi-square test, while it was 0.06 by the Fisher's exact test. The results suggested a trend that patients with genotype B HBV compared to genotype C had better SR to PEG-IFN therapy, although the difference was not significant. Results also showed that SR rate in patients with HBeAg-negative CHB (7/8 87.5%) was significantly higher than that in HBe+ CHB patients (8/21 38.1%, P < 0.05). CONCLUSION: Our results indicate that HBV genotype and HBeAg, especially the later, are main factors for predicting PEG-IFN therapy response in CHB patients. PMID- 16042880 TI - [Pedunculated (exophytic) hepatocellular carcinoma--a report of two cases]. PMID- 16042881 TI - [Hepatitis B virus genotypes and alanine aminotransferase levels in HBeAg negative patients with chronic hepatitis B and liver cirrhosis]. AB - OBJECTIVE: To investigate genotypes of the hepatitis B virus (HBV) and alanine aminotransferase (ALT) levels of HBeAg negative patients with chronic hepatitis B and liver cirrhosis. METHODS: HBV serological markers and ALT levels were detected in 62 patients with chronic hepatitis B and 41 cases with liver cirrhosis, using enzyme linked absorbent immunoassays and an enzyme method, respectively. A polymerase chain reaction of S region was used for HBV genotyping. RESULTS: Of the 62 patients with chronic hepatitis B, 21 (33.9%) were HBeAg negative, and 41 (66.1%) HBeAg positive. Among 41 cases with liver cirrhosis, 28 (68.3%) were HBeAg negative, and 13 (31.7%) HBeAg positive. Of these 62 patients with chronic hepatitis B, 53 (85.5%) were infected with HBV genotype C, and 9 (14.5%) with genotype B. Thirty-nine (95.1%) of the 41 patients with liver cirrhosis were infected with genotype C, and 2 (4.9%) with genotype B. The proportion of HBeAg negative chronic hepatitis B patients with ALT level > 40 U/L was lower than that of the HBeAg positive group (47.6% and 85.4%, respectively) (P < 0.01). The percentage of ALT levels > 40 U/L of the negative patients with liver cirrhosis was also lower as compared to that of the HBeAg positive patients, but there was no statistical difference between the two groups, because of the small sample size (P > 0.05). CONCLUSION: The proportion of HBeAg negative patients is high in the group of chronic hepatitis B and liver cirrhosis. These patients have relatively low ALT levels, and mainly have HBV genotype C infection. PMID- 16042882 TI - [Clinical investigation of famciclovir in chronic hepatitis B patients irresponsive to alpha interferon treatment]. AB - OBJECTIVES: To evaluate the efficacy and safety of famciclovir on the decreasing levels of serum HBV-DNA and ALT and HBeAg/antiHBe seroconversion in chronic hepatitis B patients irresponsive to 3 months treatment with alpha interferon. METHODS: Two hundred and nineteen patients with chronic HBV infection, defined as positive HBsAg, HBeAg and HBV DNA, were enrolled and randomly half-and- half put into famciclovir and placebo groups. The two groups received either famciclovir 500 mg tid or a placebo treatment for 24 weeks, and then were followed-up for another 24 weeks with no treatment. RESULTS: At the end of 24 weeks, the log value of HBV DNA dropped from 6.54+/-1.26 to 5.70+/-2.03 in the famciclovirt group and were elevated from 6.30+/-1.32 to 6.51+/-1.65 in the placebo group (P < 0.01). The rate of cases with persistence HBV DNA dropped 2 log of quantity in the famciclovir group and was 28.28% (28/99); it was 9.47% (9/95) in the placebo group (P < 0.01). Those with persistence negative HBV DNA was 28.28% (28/99) in the flamciclovir treated group and 14.74% (14/95) in the placebo group (P < 0.05). Those persistently being HBeAg negative were 7.69% (7/91) in the famciclovir treated group and 3.33% (3/90) in the placebo group (P > 0.05). The HBeAg/antiHBe seroconversion was 4.40% (4/91) in the famciclovir group and 2.22% (2/90) in the placebo group (P > 0.05). The percentage of cases with normal of ALT level was 15.15% in the famciclovir group and 6.35% in the placebo group (P < 0.05). CONCLUSION: Famciclovir is effective in inhibiting HBV DNA replication and in decreasing serum ALT levels. The rate of HBeAg/antiHBe seroconversion in the famciclovir treated group was similar to that of the placebo group. Famciclovir was well tolerated without severe adverse effects during our treatment. PMID- 16042883 TI - [Immune response enhanced by genes encoding IFN-alpha 8 and T alpha 1 co inoculated with HBV DNA vaccine]. AB - OBJECTIVES: To evaluate if the humoral immune response of hepatitis B DNA vaccine pVAX1-S2S could be enhanced by Talpha1 and/or IFNa expression plasmid co inoculated. METHODS: The following mammalian expression recombinant plasmids were constructed: the plasmid pVAX1-S2S expressing hepatitis B surface antigen S2S, the plasmid pVAX1-T/I co-expressing thymosin a and IFNalpha, the plasmid pVAX1 I/S2S co-expressing IFNalpha and S2S. These plasmids were inoculated intramuscularly into several BALB/c mice groups in different combinations. In the co-immunization group 1 (pVAX1-I/S2S), each mouse was inoculated with 100 microg of pVAX1-I/S2S; in the co-immunization group 2 (pVAX1-S2S) each mouse was co inoculated with pVAX1-S2S and 50 microg of pVAX1-TI; in the control group each mouse was inoculated with 100 microg of pVAX1-S2S. All the immunizations were boosted at 2 and 4 week intervals; then the serum samples were collected to detect the anti-HBs and anti-preS2 strengths. RESULTS: 3, 5 and 8 weeks after the first inoculation, the positive rates of anti-HBs were 12.5%, 12.5%, 62.5% respectively in the co-immunization group 1 and 25%, 50%, 50% in the co immunization group 2, while those in the control group were 0, 25%, 37.5%. The titers of anti-preS2 in co-immunization group 2 was 5 times higher than those in the other two groups. CONCLUSION: The data shows that Talpha1 and/or IFNalpha expression plasmid co-inoculated with pVAX1-S2S might act as an adjuvant to enhance the humoral immune response induced by pVAX1-S2S. PMID- 16042884 TI - [Establish a transgenic mice model harboring structural genes of hepatitis C virus]. AB - OBJECTIVES: To establish an animal model of HCV transgenic mice to elucidate the pathogenesis of hepatitis C virus infection and function of the viral structural proteins. METHODS: Structural gene of HCV were amplified and recombined into eukaryotic expression vectors, pcDNA4HisMax and pMT/BiP/V5-His A, after their expressive activity was confirmed to detect the structural protein in the transfected COS7 and S2 cells by Western blot. The fertilized expression element, which contained CMV or pMT promoter, structural gene of HCV and polyadenylation signal sequence, was microinjected into 1736 C57BL/6 mouse fertilized ova. The ova were then replanted into the oviducts of 69 pseudopregnant recipient mice. RESULTS: Twenty-five recipient mice were impregnated and later produced 105 newborns; 49 of them died from unknown causes and 57 survived. After the specific HCV structural genes were identified by PCR and Southern blot hybridization, 26 founders were obtained; among them 10 were stable expression mice and 16 were the inducible ones. The rate of founders developed from implanted embryos was only 1.50%. Through hybridization with normal mice, 58 hybrid mice have been obtained at present. CONCLUSION: Two kinds of different transgenic mice of HCV were developed; one is of stable expression, and the other is inducible. This transgenic mice model may create an opportunity for studying the function of the structural gene of HCV and elucidate its pathogenicity. PMID- 16042885 TI - [Molecular mechanism of HCV NS5A on p53's inhibition of AFP expression in hepatocellular carcinoma cells]. AB - OBJECTIVE: To explore hepatitis C virus (HCV) non-structural protein 5A (NS5A)'s influence on inhibition of AFP expression executed by p53 protein and its possible molecular mechanism. METHODS: Plasmid transfection and MEIA were employed to observe p53's inhibitive effect on AFP expression of Huh7 cells and the HCV NS5A's influence on p53 function. Western blot was employed to find out if HCV NS5A affects p53 protein expression and GST pull down assay was applied to examine the interaction between HCV NS5A and p53. RESULTS: The AFP concentration in the supernatant of the culture of the Huh7 cells transfected with pRc/CMV was (14322+/-2412) ng/ml, and that of the Huh7 cells transfected with pCNS5A was (13843+/-3218) ng/ml; no significant difference existed between these two groups (t = 1.42, P > 0.05). After transfection with pC53-NS3, the AFP level was decreased to (10 241+/-1326) ng/ml, and in comparison to the above two groups it had a statistically significant difference (t values were 2.41 and 2.38, P < 0.05). When co-transfected with pCNS5A and pC53-NS3, the AFP expression (14582+/ 1238) ng/ml returned to the level of pRc/CMV transfected, and there was a remarkably significant difference between this and that of the pC53-NS3 transfected cells (t = 3.12, P < 0.01). HCV NS5A had no function on the p53 protein expression with Western blot experiment. In the GST pull down assay, an HCV NS5A protein band was found after GST-p53 was added, but not detected with GST only. CONCLUSION: We found that p53 has an inhibitive function on the AFP expression in Huh7 cells and HCV NS5A minimized this p53 function. HCV NS5A did not affect p53 protein expression, but was able to form a complex with p53, by which HCV NS5A inactivated this p53 function. PMID- 16042886 TI - [Dynamic evolution of MMP-2 gene expression and its enzymatic activities in experimental liver fibrosis]. AB - OBJECTIVES: To explore the dynamic changes and interactions between MMP-2 and TIMP-2 during experimental liver fibrosis. METHODS: Wistar rats were randomly allocated into a normal group and a model group. To induce liver fibrosis, rats were injected intraperitoneally with dimethylnitrosamine (DMN) three consecutive times in the first week, then two consecutive times per week, totally for 6 weeks. In the normal control group, rats were injected with saline by the same method as the model group. Animals were sacrificed 1, 4, 10, 17, 28, 42, 56 days after starting DMN injections. Conventional histological examinations of the livers were performed with hematoxylin and eosin and Masson staining. The fibrosis was classified into 0 to 4 stages. Hydroxyproline content was determined after liver tissues were hydrolyzed in HCl at 160 degree C for 2 hrs and then measured with spectrometry at 560 nm wavelength. mRNA levels of MMP-2 and TIMP-2 were determined by semi-quantitive RT-PCR. Gelatinase activity of MMP-2 was examined by zymography using gelatin substrate. RESULTS: In the model group the hepatic MMP-2 mRNA expression started to increase 10 days after DMN administration and remained at a much higher level than in the normal group throughout the study period, while TIMP-2 mRNA expression started to be lower than in the normal group 17 days after DMN administration and reached the lowest level on the 28th day. Then it rapidly rebounded and remained higher than that in the normal group from the 42nd day to the end of the study period. TIMP-2/MMP-2 began to be lower by several days than that of the normal group after DMN administration through the remaining study period. Zymography showed that the enzymatic activities of both latent MMP-2 and active MMP-2 were increased during the process of liver fibrosis. CONCLUSION: In liver fibrosis, MMP-2 expression increases, while TIMP-2 expression relatively decreases. The enzymatic activities of MMP-2 increase as the liver fibrosis develops. PMID- 16042887 TI - [Correlation between HBsAg, HCV expression in cancerous and pericancer tissues of hepatocellular carcinoma and liver fibrosis markers in serum]. AB - OBJECTIVE: To study the correlation between HBsAg and HCV in hepatocellular carcinoma (HCC) pericarcinomatous tissues and serum liver fibrosis markers. METHODS: The patterns of HBsAg and HCV in 100 cases of HCC and their surrounding liver tissues were studied with paraffin sections using immunohistochemistry techniques. Hyaluronic acid (HA), procollagen III peptide (PIIIP), collagen IV (CIV), and laminin (LN) were detected by radioimmunoassay. RESULTS: The levels of HA, PIIIP, CIV and LN in the HBV and HCV coinfection group were the highest among the four groups. The levels of HA, PIIIP, CIV and LN in the groups not infected by HBV and HCV were the lowest among the four groups. HBV and HCV expressions were positively correlated with HA, LN and CIV and their Spearman's rank correlation coefficients were 0.60, 0.45, 0.46, respectively (P < 0.01). CONCLUSION: Liver cancer development follows a sequential trend of chronic hepatitis, liver cirrhosis, and liver cancer. In the tissues of liver cancer with virus infection background, the serum marker level of hepatic fibrosis is obviously higher than those without virus infection background. On the one hand, virus infection is one of the reasons causing liver cancer; and on the other hand, longstanding viremia will aggravate pathological changes of liver tissues. Therefore antivirus treatment of hepatitis is of significance for the prognosis of liver cancer. PMID- 16042888 TI - [Pathological studies of tissue damage in experimental rabbit liver cancer induced by energy controllable steep pulses]. AB - OBJECTIVE: To evaluate the efficacy of energy controllable steep pulses (ECSP) in the treatment of rabbit VX2 cancer implanted in livers. METHODS: A tumor model was successfully established using 30 rabbits. ECSP were applied to liver cancer in half of these rabbits and the rest were used as controls. After exposure to ECSP, tissues were obtained and subjected by routine HE and transmission electron microscopic (TEM) observation. The survival time of the animals and the statuses of each group were recorded. RESULTS: From pathological observations, ECSP showed effectively destructive action compared with that of the unexposed group. A clear borderline can be seen between necrotic cancer and its surrounding normal tissue. Irreversible cell changes were present under TEM. The survival periods of the experimental and control group were 83.1 days and 39.0 days respectively, and there was a significant difference between the two groups (Z = -2.943, P < 0.01). CONCLUSION: ECSP can effectively treat rabbit VX2 cancer implanted in the liver; also it is safe for its surrounding normal tissues. ECSP can be a useful method for local treatment of liver cancer. PMID- 16042889 TI - [Screening of binding proteins to interferon-alpha promoter DNA by phage display technique]. AB - OBJECTIVE: To investigate the interferon alpha regulation mechanisms by screening binding proteins of interferon alpha promoter by phage display. METHODS: PCR product of interferon-alpha promoter was incubated with a phage display cDNA library that expressed a library of human liver proteins on the surface of bacteriophage T7. Unbound phages were washed off and the phages bound to the interferon alpha promoter were amplified. Positive plaques were amplified by PCR and cloned into a pGEM-Teasy vector in order to perform DNA sequence analysis and subsequent computer blasting analysis. RESULTS: Positive phage-displayed proteins binding with interferon alpha promoter were enriched after five rounds of biopanning. We found that the following proteins were relevant to interferon alpha: mitochondrial ribosomal protein, chromosome clone, fibrinogen A alpha polypeptide, enoyl coenzyme A hydratase short chain, eukaryotic translation elongation factor 1 alpha, PI-3-kinase-related kinase SMG-1-like, xeroderma pigmentosum C group, and Homo sapien activity-dependent neuroprotector (ADNP). CONCLUSION: Many proteins with different functions could bind with interferon alpha promoter. PMID- 16042890 TI - [The heterogeneity of HCV quasi-species in the hypervariable region and its relationship to interferon therapy]. PMID- 16042891 TI - [Evaluation of early stage diffused liver lesions with MR functional diffusion weighted imaging--an experimental study]. AB - OBJECTIVE: To investigate the value of Diffusion-Weighted Imaging (DWI) in the diagnosis of early stage liver diffuse lesions. METHODS: Diethylnitrosamine (DEN) was used to induce liver lesions in rats. Sequential DWI studies were performed on the livers from 1 to 14 weeks after DEN was administered through drinking water. Comparing studies with a blank control group was set and pathohistological examinations of the livers were performed. RESULTS: No obvious routine MRI morphological change was found in either group during this period, but DWI demonstrated heterogeneous changes in the test group at the cirrhosis stage. There was no significant alteration of the apparent diffusion coefficient (ADC) value in the control group during this period (P > 0.05). The ADC values of the test group began to decline from the fifth week. Until the tenth week, the ADC value of the test group decreased drastically and when b = 300 s/mm2 statistic, the results showed an obvious difference between the two groups. There were also differences between the ADC values at the 10th, the 9th and the 1st weeks of the test group (P < 0.05). When b = 600 s/mm2 and 1000 s/mm2, significant differences were found after the sixth week between the two groups (P < 0.01). The main pathohistological liver change in the test group during the 1 to 4 week period after DEN was administered was swelling of hepatocytes; during the 5 to 8 week period it was fibrous tissues hyperplasia, and in the 9 to 14 week period it was cirrhotic nodule formation. CONCLUSION: MR functional DWI could detect liver diffuse lesions earlier than conventional MR imaging. Measurement of ADC value may be of use in early diagnosis of liver diffuse diseases and for monitoring the changes of the lesions. PMID- 16042892 TI - [Effects of gamma-interferon on gene expression of collagen I, III and on the tissue inhibitor of metalloprotenase 1 in HSC-T6 cells]. AB - OBJECTIVE: To observe the in vitro effects of gamma-interferon (IFNgamma) on gene expression of collagen I (Col I), III (Col III) and tissue inhibitor of metalloprotenase 1 (TIMP1) of HSC-T6 cells. METHODS: Cultured HSC-T6 cells were exposed to IFNgamma at concentrations of 0.1, 1, 10, 10(2), 10(3), 10(4), 2.5 x 10(5), 5 x10(5) U/ml for 48 hours. 4,5-simethylthiazaoly colormetric assay was used to evaluate the effect of IFNgamma on HSC-T6 cell proliferation. After incubating with IFNgamma (1 U/ml, 10(2) U/ml and 10(4) U/ml) for 48 hours, HSC-T6 cells were harvested to detect Col I, Col III and TIMP1 steady state mRNA levels by quantitative reverse-transcription polymerase chain reaction (RT-PCR). RESULTS: The Col I, Col III and TIMP1 mRNA levels of the control group were 2.86+/-0.21, 2.00+/-0.23 and 3.90+/-0.81, respectively. Col I and Col III mRNA levels in HSC-T6 cells treated by different concentrations of IFNgamma were lower than that of the controls (P < 0.01). There was no significant difference in TIMP1 mRNA levels between IFNgamma groups and controls. CONCLUSION: IFNgamma suppresses expression of Col I and Col III whereas it has no effect on TIMP1 mRNA expression. The antifibrotic mechanism of IFNgamma may be partly due to its down regulation of Col I and Col III mRNA levels in HSC-T6 cells. PMID- 16042893 TI - [The changes of ROS and mitochondria membrane potential in HepG2 cells on the pressure of cisplatin]. AB - OBJECTIVE: To explore the changes and significance of the level of reactive oxygen species (ROS) and mitochondria membrane potential (Delta Psi) in HepG2 cells under the stress of cisplatin (CDDP). METHODS: HepG2 cells were incubated with CDDP. The changes in the level of ROS were determined by a probe (2,7 dichloro fluorescein-ciactate, DCFH-DA) and the changes of Delta Psi were reflected as changes of intensities of fluorescence seen under a laser scan microscope using a probe (rhodamine-123). All these changes in cells at 0 h, 24 h, 48 h, 72 h, 120 h, 168 h were dynamically observed. RESULTS: The level of ROS was much higher after the CDDP treatment than the non-treated, and the increase lasted for 24 h and 48 h. Then it started to decrease at 72 h, gradually returning to normal level at 120 h. Under the selective pressure of CDDP, the fluorescence intensity of rhodamine-123 in HepG2 cells was decreasing at 24 h and 48 h, then gradually started to increase at 72 h. There were no such changes in the cells of the controls. CONCLUSION: The changes of ROS and Delta Psi in HepG2 cells under the pressure of CDDP suggest that the cells change themselves adapting to such pressures. PMID- 16042894 TI - [Durability of HBeAg seroconversion in lamivudine treatment of chronic hepatitis B patients]. AB - OBJECTIVE: To investigate the factors which may affect the rate of HBeAg seroconversion and its durability after long-term lamivudine therapy in chronic hepatitis B patients. METHODS: 81 patients were treated in a phase III clinical trial with lamivudine 100 mg daily for up to 5 years. The mean period of treatment was (48.84+/-10.52) months (range: 16 approximately 60 months). When HBeAg seroconversion occurred in the patients, which was defined as loss of HBeAg and detection of anti-HBe antibody, HBV DNA level less than 10 mEq/ml more than two times (once every 3 months), the lamivudine treatment was stopped and they were followed-up for another 6 approximately 12 months. The HBV DNA level was detected using Branched DNA assay (Chiron). The HBV markers were detected using IMX assay (Abbott). HBV genotyping was performed using type-specific PCR. The data were analyzed using logistic multivariant analysis. RESULTS: (1) The distribution of HBV genotypes was as follows: type B, 17 (20.97%), type C, 62 (76.54%), and type B+C, 2 (2.47%). (2) 26 patients achieved HBeAg seroconversion (32.10%). The annual seroconversion rates were 16.05% (13/81) in the 1st year, 19.75% (16/81) in the 2nd, 27.16 % (22/81) in the 3rd, 28.40% (23/81) in the 4th and 32.10% (26/81) in the 5th year. Four patients had a reappearance of HBeAg and an elevation of HBV DNA. Therefore the stability ratio was 84.62% (22/26). The mean baseline ALT and HBV DNA levels in those who were seroconvered were (104.8+/ 86.3) U/L and (940.1+/-1123.7) mEq/ml, respectively. Mean baseline ALT and HBV DNA of non-seroconverters were (48.3+/-46.9) U/L and (2152.3+/-3063.5) mEq/ml. There was a significant difference between the two groups shown by Kruskal-Wallis Test (P < 0.05). Analysis by logistic multivariate analysis showed that the rate of HBeAg seroconversion and its durability rate correlated with a high baseline ALT. In contrast, a relatively low seroconversion rate and durability rate was observed in patients with high baseline HBV DNA. The durability rate also correlated with additional lamivudine treatment after HBeAg seroconversion. CONCLUSION: Continuation of lamivudine therapy for more than 6 months after HBeAg seroconversion might increase the durability of response. PMID- 16042895 TI - [Some strategies for the long-term treatment of chronic hepatitis B]. PMID- 16042896 TI - [Management of HBeAg negative chronic hepatitis B]. PMID- 16042897 TI - [Antiviral treatment of HBeAg positive chronic hepatitis B]. PMID- 16042898 TI - [Inhibitory effects of a novel trimeric of beta peptide (beta3) on migration and invasion of hepatocellular carcinoma cells]. PMID- 16042899 TI - [The sensitivity improvement of HBx+HepG2 to CDDP with PD98059]. PMID- 16042900 TI - [Effects of hyperbaric oxygen with free-radical antagonists on the expression and activity of matrix metalloproteinase-2 in rat livers]. PMID- 16042901 TI - [Effects of selective cyclooxygenase-2 inhibitor NS-398 on proliferation and cell cycle progression of hepatocarcinoma cells]. PMID- 16042902 TI - [The characterization value of feridex-enhanced MR imaging for focal hepatic lesions in reference to their histological pathology]. PMID- 16042903 TI - [Expression of IL-10 in the livers of rats with chronic alcoholic liver diseases]. PMID- 16042904 TI - [Effects of herbal compound 861 on the ultrastructure and cytoskeleton protein expression of hepatic stellate cells]. PMID- 16042905 TI - [A report from the Third National Symposium on Liver Failure and Artificial Livers]. PMID- 16042906 TI - [L-carnitine in the treatment of liver diseases]. PMID- 16042907 TI - Diagnosis and treatment of irritable bowel syndrome: state of the art. AB - Irritable bowel syndrome (IBS) is a common functional gastrointestinal disorder with a wide variety of presentations that may include abdominal pain, bloating, diarrhea, constipation, or alternating bowel habits. Symptom-based criteria and a limited medical evaluation are used for diagnosis. The heterogeneity of IBS presenting symptoms, together with the pathophysiology of the disorder, is unclear, making treatment challenging. Treatment strategies are focused on specific symptoms, potential underlying disorders in stress responsiveness, and predisposing psychological features. Although only two medications, tegaserod for constipation-predominant IBS and alosetron for diarrhea-predominant IBS, are specifically indicated, a wide variety of treatment options are available and are discussed in this review. PMID- 16042908 TI - The role of gender and biological sex in irritable bowel syndrome. AB - Irritable bowel syndrome (IBS), which is characterized by abdominal pain or discomfort that is associated with altered bowel function (diarrhea, constipation, or alteration between the two), is one of several gastrointestinal motility disorders. IBS affects up to one in five North Americans, mostly women. The reason(s) this disorder is reported more often by women than men, and the role of gender and biological sex in the prevalence, pathophysiology, symptom presentation, impact on quality of life, diagnosis, and response to treatment, are poorly understood. The purpose of this article is to review the evidence surrounding the roles of gender and biological sex in IBS. PMID- 16042909 TI - Symptom overlap and comorbidity of irritable bowel syndrome with other conditions. AB - Irritable bowel syndrome (IBS) is one of several highly prevalent, multi-symptom gastrointestinal motility disorders that have a wide clinical spectrum and are associated with symptoms of gastrointestinal dysmotility and visceral hypersensitivity. Symptom overlap and comorbidity between IBS and other gastrointestinal motility disorders (eg, chronic constipation, functional dyspepsia, gastroesophageal reflux disease), with gastrointestinal disorders that are not related to motility (eg, celiac disease, lactose intolerance), and with somatic conditions (eg, fibromyalgia, chronic fatigue syndrome), are frequent. The clinical associations and pathophysiologic links between IBS and these disorders continue to be explored. This review discusses overlapping symptoms and comorbidity of IBS with select gastrointestinal and non-gastrointestinal disorders and attempts to identify commonalities among these conditions. PMID- 16042910 TI - New insights into the pathophysiology of irritable bowel syndrome: implications for future treatments. AB - Irritable bowel syndrome (IBS) is a multifactorial disorder characterized by abdominal pain and altered bowel habits. Chronic symptoms may occur due to changes in gastrointestinal motor function, enhanced perception of gut stimuli, and psychosocial factors. Recent data suggest that abnormal processing of afferent signals occurs in IBS patients. A newly recognized causative factor in a subset of IBS patients is post-infectious IBS. Altered transport of intestinal gas and bowel distention may contribute to abdominal discomfort, pain, and bloating. Changes in gut microflora have also been reported, but data remain scant. Advances have been made in our understanding of serotonin signaling and metabolism in IBS patients, in part due to the introduction of specific receptor agonists and antagonists. Finally, exciting data are emerging on genetic alterations that may contribute to the pathophysiology and treatment of IBS. Increasingly novel mechanisms are being identified that should aid in better understanding of the complex pathophysiology of IBS and developing new therapies. PMID- 16042911 TI - Sickness and satiety: physiological mechanisms underlying perceptions of nausea and stomach fullness. AB - The pathophysiology of nausea and the physiological mechanisms underlying perceptions of stomach emptiness and fullness are not clearly understood, but several potentially important factors have been identified. Gastric dysrhythmias are believed to contribute to the subjective experience of nausea and may also be involved with perceptions of stomach emptiness, hunger, and even dyspepsia symptoms like bloating and early satiety. Normal gastric neuromuscular function is more evident in the absence of nausea and is also thought to be related to feelings of satiety or comfortable stomach fullness. Autonomic and endocrine influences may also play a critical role in the pathophysiology of nausea and abnormal perceptions of stomach emptiness or fullness. Achieving a better understanding of the gastric neuromuscular and neurohormonal influences on perceptions arising from the viscera may prove invaluable in the development of novel treatments for such conditions as unexplained nausea, functional dyspepsia, and obesity. PMID- 16042913 TI - Dietary intake and the risk of GERD. PMID- 16042914 TI - Nasogastric versus nasojejunal feeding in severe acute pancreatitis. PMID- 16042912 TI - Frontiers in functional dyspepsia. AB - Functional dyspepsia (FD) refers to unexplained pain or discomfort in the upper abdomen and is commonly seen in gastroenterology practice. The underlying pathophysiologic mechanisms associated with FD are unclear, although traditionally, delayed gastric emptying, visceral hypersensitivity to acid or mechanical distention, and impaired gastric accommodation have been implicated as putative physiologic disturbances. It also remains uncertain whether FD and irritable bowel syndrome are different presentations of the same disorder. Recent data on pathophysiologic mechanisms of FD have focused on postprandial motor disturbances (accelerated gastric emptying, antral-fundic incoordination, and abnormal phasic contractions), alterations of neurohormonal mechanisms in response to a meal, and previous acute infection. Pharmacologic therapies for FD may be guided by these novel mechanisms, as current available therapeutic options are limited. Novel prokinetics and gastric accommodation modulators, visceral analgesics, and agents targeting the neurohormonal response to food ingestion are the next therapeutic frontiers in FD. This review summarizes traditional knowledge and more recent advances in the pathophysiology of FD and potential therapeutic opportunities. PMID- 16042915 TI - Herbal supplements. AB - Herbal dietary supplements are taken by many individuals, often without the knowledge of their clinicians. Herbals have many pharmacologic and medicinal properties but are unregulated with respect to manufacturing purity, safety, and efficacy. Although clinicians often dismiss the utility of herbal therapy, many randomized controlled trials of herbal use in gastrointestinal and liver diseases have been conducted. There is no evidence for benefit from herbal supplements in the treatment of liver disease, but sufficient evidence exists for their use in functional gastrointestinal symptoms to warrant further investigation. PMID- 16042916 TI - Clinical implications of oxidative stress and antioxidant therapy. AB - Oxidative stress occurs when there is an imbalance between generation of reactive oxygen species and inadequate antioxidant defense systems. Oxidative stress can cause cell damage either directly or through altering signaling pathways. Oxidative stress is a unifying mechanism of injury in many types of disease processes, including gastrointestinal diseases. For example, in alcoholic liver disease, reactive oxygen species have been detected through direct spin-trapping techniques and through indirect markers, such as products of lipid peroxidation. A host of antioxidants have protected against liver injury in animal models of alcoholic liver disease. Similarly, in inflammatory bowel disease, oxidative stress has been postulated to play a role in disease initiation and progression, and antioxidant therapy, such as green tea polyphenols and gene therapy with superoxide dismutase, has a markedly attenuated disease. Downregulation of specific detoxification genes may play a role in the pathogenesis of inflammatory bowel disease, especially in ulcerative colitis. Oxidative stress is postulated to play a sustaining role in acute and chronic pancreatitis. Antioxidant supplementation has been used with some success in the treatment of chronic pancreatitis. This review covers recent findings related to oxidative stress in liver disease, inflammatory bowel disease, and pancreatitis. PMID- 16042917 TI - Management of small intestinal bacterial overgrowth. AB - Similar to that of all mammals, the human gastrointestinal tract is colonized by 100 trillion bacteria shortly after birth. Remarkably, in the open-tube arrangement of the intestine, this bacterial population is tightly compartmentalized to the distal gut. Contamination of the small intestine with colonic bacterial flora or small intestinal bacterial overgrowth (SIBO) has been understood previously as a complication of uncommon conditions associated with obvious intestinal stasis. However, SIBO has also been found in 78% to 84% of patients with the common condition of irritable bowel syndrome (IBS). In this paper, the diagnostic and treatment approaches to SIBO are reconsidered within the larger framework of the patient with IBS. PMID- 16042918 TI - Gastrointestinal complications of bariatric surgery. AB - The current epidemic of obesity and its myriad comorbidities represents one of the true major public health crises in the United States today. Data reflecting durable weight loss achieved through surgical treatment of obesity demonstrate impressive results heretofore unattainable with less aggressive modalities. However, these outcomes are accompanied by expected side effects and complications, which require specific knowledge to diagnose accurately and a specialized skill-set to manage precisely. With the ever-increasing demand for bariatric surgery, the number of ensuing complications will also rise, requiring the expertise of a gastroenterologist for both diagnosis and treatment. Thus, the gastroenterologist must develop a keen understanding of bariatric surgical anatomy and physiology, as well as the expected postoperative side effects and potential complications experienced by the post-bariatric patient. PMID- 16042919 TI - Nutritional support of the obese patient. AB - Overweight and obesity is the most common chronic disease in the United States and is associated with an increased risk for morbidity and mortality. For the hospitalized patient, the mechanical, metabolic, and inflammatory physiologic changes induced by obesity necessitate additional considerations for care. Calculation of nutritional requirements is problematic and challenging due to difficulty in measuring body composition and energy expenditure. Provision of selective hypocaloric feeding in this population may be particularly beneficial in reducing complications of hyperglycemia, fluid overload, and reduction in fat mass. Clinical care should incorporate a team approach that addresses the special nutritional and metabolic needs of this population. PMID- 16042921 TI - Introduction to flap movement: reconstruction of five similar nasal defects using different flaps. AB - BACKGROUND: There are several options for closure of a given surgical defect after tumor extirpation is confirmed. Flap reconstruction is one of these options. OBJECTIVE: The purpose of this article is to introduce the three basic types of flap movement: advancement, rotation, and transposition. METHODS: Five similar defects located on the nasal sidewall were repaired, each using a different flap design. RESULTS: The optimal flap design for a given defect on a particular patient is based on the answers to a series of questions: Where is the available tissue reservoir? How can tissue be mobilized from the reservoir to cover the defect? How do the resulting tension vectors affect critical structures? Where are the final incision lines? CONCLUSION: Many factors must be evaluated before determining a method of reconstruction. Flap reconstruction requires a thorough understanding of anatomy and tissue movement. PMID- 16042922 TI - Advancement flaps: a basic theme with many variations. AB - BACKGROUND: The advancement flap involves the linear advancement of tissue in one direction. Despite its straightforwardness and simple concept, it can be used to close a variety of defects, ranging from small defects on the scalp or extremities to large, complicated defects involving cosmetic units on the face. OBJECTIVES: To provide a basic and useful review for the indications, advantages, disadvantages, and techniques for the use of advancement flaps in the reconstruction of defects in dermatologic surgery. MATERIALS AND METHODS: We performed a literature search for articles discussing advancement flaps and compiled a brief review of our findings. RESULTS: The movement of the advancement flap must be balanced by the blood supply of the flap. The excision of Burow's triangles along various aspects of the advancement flap can increase movement and improve cosmesis of the flap. The types of advancement flaps discussed include the single advancement flap, double advancement flap, A-T flap (O-T flap), Burow's triangle flap (Burow's wedge flap), crescenteric advancement flap, island pedicle flap (V-Y flap), helical rim advancement flap, and facelift flap. CONCLUSION: Advancement flaps are versatile and useful basic flaps for repairing defects. PMID- 16042923 TI - The island pedicle flap. AB - BACKGROUND: Surgical defects following the extirpation of cutaneous neoplasia may present challenges to the dermatologic surgeon. There are many repair options for an individual defect, including second-intention healing, primary closure, skin graft, and skin flap closure. The island pedicle flap is a random pattern advancement flap well suited to reconstruct a variety of small- to intermediate sized soft tissue wounds. OBJECTIVE: To review the utility of the island pedicle flap in reconstructive dermatologic surgery and to detail the operative technique to achieve reproducible functional and esthetic results. METHODS: The method of flap reconstruction is presented. A variety of defects repaired with this technique are reviewed. RESULTS: The subcutaneous island pedicle flap yielded excellent functional and cosmetic repair of the presented defects with minimal operative morbidity. Secondary to a well-preserved central vascular pedicle, complications are infrequent. CONCLUSIONS: The random pattern island pedicle flap is a versatile and robust flap used to repair a variety of soft tissue wounds in a single-stage procedure with reproducible operative outcomes. PMID- 16042924 TI - Rotation flaps. AB - BACKGROUND: Rotation flaps are arcuate repairs that redistribute tension vectors and recruit adjacent and/or distant tissue laxity. Rotation allows for the closure of wounds that cannot be repaired along a single tension vector. A rich and evolving literature details the evolution of rotation as an elegant method of repair for surgical wounds. OBJECTIVE: The goal of this article is to understand the basic principles and proper execution of tissue rotation for the repair of facial operative wounds, with special attention given to the concept of pivotal restraint and with a step-by-step regional approach. METHODS AND MATERIALS: A review of the literature of dermatologic surgery, plastic surgery, and otolaryngology leads to a detailed understanding of rotation flap design and execution. RESULTS: Proper rotation flap design allows for the closure of large and complex wounds that will not close along one motion while minimizing tension vectors that affect adjacent free margins. CONCLUSIONS: The concept and execution of rotation are integral to the practice of dermatologic surgery. Proper design and undermining are essential to create an adequately sized flap and to free pivotal restraint to facilitate wound closure. In many cases, the arc of a rotation flap may be hidden within a natural cosmetic boundary, allowing for an elegant and minimally visible reconstruction. PMID- 16042925 TI - Transposition flaps in cutaneous surgery. AB - BACKGROUND: Transposition flaps are powerful reconstructive tools that are frequently called on in cutaneous reconstruction. Transposition flaps tap into adjacent areas that may have relative laxity while redirecting the vectors of tension during closure. OBJECTIVE: As with the closure of any surgical defect, the goal is to achieve the best possible functional and esthetic result. MATERIALS AND METHODS: Manual manipulation is used to "feel" for areas of relative laxity and test the effects of various tension vectors on adjacent structures. Every effort should be made to avoid distortion of the free margins of structures such as the nose and eyelids. Consideration must be taken to best camouflage incision lines within existing lines or creases, at the junction of cosmetic units, or at least parallel to lines of relaxed skin tension. CONCLUSIONS: A complete knowledge of the possible variations and modifications of transposition flaps can help fine-tune the execution of the flap to provide the patient with the best possible result. Good surgical technique and proper wound eversion through meticulous suture placement also help tremendously in consistently attaining esthetically pleasing results. At the surgical bedside, an artistic eye should meet the science of cutaneous biomechanics. PMID- 16042926 TI - Reconstructive utility of the bilobed flap: lessons from flap successes and failures. AB - BACKGROUND: Although the bilobed transposition flap's utility in the reconstruction of difficult facial surgical wounds is often praised, the complicated design of the flap can produce unacceptable esthetic results if the nuances of flap design and execution are not properly understood. OBJECTIVE: To review the historical development of the bilobed flap and to offer contemporary suggestions on how to manipulate the flap's design to improve the flap's performance. METHODS: Illustrative case examples of bilobed flap successes and failures are appropriately and thoroughly examined. RESULTS: The proper use of the bilobed flap completely depends on understanding the design modifications of the flap that can impact the flap's final appearance. CONCLUSIONS: The bilobed flap, with its complicated design and requirement of meticulous operative technique, introduces tremendous opportunities for esthetic misadventure. Nonetheless, the flap, when properly constructed, deserves its reputation as the reconstructive flap of choice for many difficult wounds, particularly those wounds located on the distal nose. PMID- 16042927 TI - Staged cheek-to-nose and auricular interpolation flaps. AB - BACKGROUND: Staged interpolation flaps are priceless options in skin cancer reconstruction. Their value lies in their flexibility, reach, reliability, and ability to repair distant, complex facial defects. Familiar interpolation flaps to dermatologic surgeons include the paramedian forehead flap, cheek-to-nose interpolation flaps, and auricular staged flaps. OBJECTIVE: In this special reconstructive issue, the paramedian forehead flap is discussed separately. This article highlights the cheek-to-nose and auricular interpolation flaps as applied to skin cancer defects. Design considerations, anatomic basis, execution, and the distinctions of each repair are presented. MATERIALS AND METHODS: Patients with facial defects from Mohs micrographic surgery serve to illustrate the surgical techniques of each repair. RESULTS: With meticulous planning and thoughtful execution, cheek-to-nose and auricular staged flaps are capable of restoring both function and cosmesis. Several surgical stages are necessary, and an adequate supporting infrastructure is essential for an optimal outcome. CONCLUSION: Skin cancer patients with complex facial wounds from Mohs micrographic surgery may be assured of the highest possible cure rate. Further, their esthetic and functional reconstructive goals may be achieved with staged flaps for the nose and ear. PMID- 16042928 TI - Paramedian forehead flap reconstruction for nasal defects. AB - BACKGROUND: The paramedian forehead flap is the ultimate reconstructive method for repair of extensive nasal defects. Changes in technique have resulted in the evolution of the modern-day forehead flap, which is a streamlined, efficient, reliable flap that can be counted on to provide superior function and cosmesis in the reconstruction of large nasal defects. OBJECTIVE: Consistent success in the execution of a forehead flap hinges on a careful stepwise approach to the defect, the patient, and the surgical technique. Characterization of these steps was undertaken to assist the surgeon in achieving consistent post- operative results. METHODS: The process of executing a paramedian forehead flap beginning with preoperative assessment through the intraoperative procedure and culminating in the postoperative care is elucidated and discussed. RESULTS: Through thoughtful planning and correct execution of technique, very large nasal defects are reconstructed, with excellent functional and esthetic results. Specific examples illustrate the range of approaches that can be used to address a variety of nasal tissue loss. CONCLUSION: With careful attention to the reconstruction of all components of a nasal defect, a forehead flap can restore virtually any large nasal defect with excellent functional and cosmetic results. The skill sets that help optimize the process of nasal reconstruction are important to acquire. With careful planning and surgical finesse, forehead flaps can often result in nearly imperceptible restoration of the nose. PMID- 16042929 TI - Repair of defects on nasal sebaceous skin. AB - BACKGROUND: Reconstructive procedures performed on sebaceous nasal skin are prone to partial flap necrosis, scar spread and inversion, and tissue mismatch. An ideal repair would optimize vascular integrity, minimize closure tension, and use adjacent tissue. OBJECTIVE: The purpose of this article is to describe a flap design and dynamics that permit satisfactory reconstruction of small- to medium sized defects on nasal sebaceous tissue. METHODS: A modified advancement flap is described that may be used on central and off-midline defects of the nasal tip. RESULTS: Use of the modified advancement flap resulted in good cosmetic results with few adverse postsurgical events. CONCLUSIONS: The modified advancement flap satisfies the requirements of a hardy blood supply, minimization of closure tension, and use of adjacent tissue. The surgical results are predictable and rarely associated with complications. PMID- 16042930 TI - Grafts in dermatologic surgery: review and update on full- and split-thickness skin grafts, free cartilage grafts, and composite grafts. AB - BACKGROUND: Skin grafting has evolved in the past centuries to encompass numerous well-established reconstruction techniques that are uniquely able to restore structure, function, and cosmesis to a variety of surgical wounds. OBJECTIVE: To provide a detailed overview of the general principles of skin grafting geared for the dermatologist and the dermatologic surgeon. METHODS: Comprehensive review of the literature. RESULTS: A summary of the different applications and techniques of full- and split-thickness skin grafts, free cartilage grafts, and composite grafts is presented. Indications, advantages, disadvantages, techniques, and complications are discussed in depth. CONCLUSION: Skin grafting is a dynamic and versatile method of cutaneous reconstruction that can be accomplished successfully with a thorough understanding of the principles and techniques of grafting. PMID- 16042931 TI - Update on the use of antibiotics in cutaneous surgery. AB - BACKGROUND: Antibiotics are often prescribed in perioperative settings, including dermatologic surgery. Given the continued evolution in the breadth and complexity of cutaneous procedures performed and inevitable localized or distant infections that occasionally occur, it seems prudent to periodically evaluate findings and recommendations from the literature regarding the use of antibiotics in cutaneous surgery. MATERIALS AND METHODS: Literature review from English-language sources from the past 30 years, especially focusing on sources from the past 5 to 10 years. Data were examined for a variety of cutaneous surgical procedures, routes of antibiotic administration, and consideration of both cutaneous and distant infections. RESULTS: The literature suggests that, for most routine skin procedures, antibiotic use is probably not warranted for the prevention of surgical wound infection, endocarditis, and late prosthetic joint infections. During prolonged Mohs procedures, delayed repairs, grafts, takedowns of interpolation flaps, or any procedure that breaches a mucosal surface, the evidence is less clear, and decisions should be made on a case-by-case basis. Topical antibiotics are probably overused, although silver sulfadiazine may have an undeserved negative reputation among dermatologists. Systemic prophylactic antibiotics for laser resurfacing and liposuction appear not to be routinely necessary, although patients with known prior herpes infection likely should receive antiviral prophylaxis. The overall reported infection rates and infectious complications remain low in dermatologic surgery, and antibiotic therapy may be prescribed excessively or inappropriately as a result. CONCLUSION.: Antibiotics continue to be widely used, and through varying routes of administration, in the setting of dermatologic surgery. Prudent use of these agents is indicated in high-risk patients, certain anatomic locations, and the presence of overt infection. Additional studies may help clarify the most appropriate indications, and in which patient populations, in the future. PMID- 16042932 TI - Use of hyaluronidase in the treatment of granulomatous hyaluronic acid reactions or unwanted hyaluronic acid misplacement. AB - BACKGROUND: In the past, reactions or misplacement of soft tissue fillers has been fraught with anxiety because time has been the main thrust for improvement in spite of ancillary treatments. Hyaluronidase is an enzyme that dissolves hyaluronic acid in the skin and also assists in the management of granulomatous foreign-body reactions to hyaluronic acid. These reactions may be caused by allergy to the material or immunologic response to the protein contaminants in the hyaluronic acid preparations. Dissolution of material in erroneous placement of material and in allergic reactions can be a time saver and a deterrent to patient dissatisfaction. OBJECTIVE: To evaluate the use of hyaluronidase in the treatment of both allergic reactions and the erroneous misplacement of hyaluronic acid in the skin. METHODS: A case of persistent granulomatous reaction to injectable hyaluronic acid and a case of hyaluronic acid erroneous misplacement with their successful subsequent treatments using intracutaneous hyaluronidase are reported, along with illustrative examples of hyaluronidase use. RESULTS: The use of hyaluronidase reduced the patient discomfort within 24 to 48 hours, deterring any patient anxiety or patient dissatisfaction. CONCLUSIONS: Hyaluronidase has a place in the treatment of allergic reactions to hyaluronic acid [corrected] and in the erroneous misplacement of the material. PMID- 16042933 TI - Long-pulsed dye laser treatment for facial telangiectasias and erythema: evaluation of a single purpuric pass versus multiple subpurpuric passes. AB - BACKGROUND AND OBJECTIVE: Subpurpuric treatments with the pulsed dye laser can be effective for treatment of vascular lesions, although less so than when purpuric fluences are used. Increased efficacy may be achieved by performing multiple passes at the time of treatment. We performed a split-face bilateral paired comparison of multiple low-fluence subpurpuric passes compared with a single high fluence purpuric pass in the treatment of facial telangiectasias. MATERIALS AND METHODS: Nine patients were included in the study. One cheek was chosen to be treated with four passes of a nonpurpuric fluence, and the contralateral cheek was treated with a single purpuric pass. Reductions in vessel density, diameter, arborization, and background erythema were evaluated 3 weeks after treatment. RESULTS: We found a 43.4% reduction in surface area covered by telangiectasias on the cheek treated with a single purpuric pass compared with 35.9% on the cheek treated with four subpurpuric passes. The purpuric fluences produced greater reduction in vessel diameter and arborization, whereas the subpurpuric protocol was more effective in reducing background erythema. Purpuric fluences were also noted to produce more significant edema and transient hyperpigmentation in one patient. CONCLUSION: The multipass subpurpuric approach to treatment with the pulsed dye laser is both cosmetically acceptable and effective, although purpuric treatments may be required to effectively eliminate larger-caliber, more highly networked vessels. PMID- 16042934 TI - Comparison of a 1,064 nm laser and a 1,320 nm laser for the nonablative treatment of acne scars. AB - BACKGROUND: There have been many reports of the use of nonablative lasers for the treatment of acne scars. OBJECTIVE: To evaluate the ability of the 1,064 nm neodymium:yttrium-aluminum-garnet (Nd:YAG) laser to treat acne scars and compare it with that of the 1,320 nm Nd:YAG laser. METHODS: Twelve patients with Fitzpatrick skin types I to III were randomly selected to have half of the face or back treated with the Lyra 1,064 nm Nd:YAG laser (Laserscope Corporation, San Jose, CA, USA) and the other half with the CoolTouch II 1,320 nm Nd:YAG laser (ICN Pharmaceuticals, Inc., Costa Mesa, CA, USA). Three treatments at 4-week intervals were performed. Patients were evaluated by photographic and profilometric methods before and 6 months after the last treatment. RESULTS: Immediate changes included mild erythema with the 1,064 nm Nd:YAG laser and mild edema and erythema with the 1,320 nm Nd:YAG laser. No long-term adverse changes were seen with either laser system. Using the 1,320 nm system, 42% of the patients had 30 to 40% clinical improvement, 42% had 11 to 29%, and 16% had 10% or less. With the 1,064 nm system, 58% had 30 to 40% clinical improvement and 42% had 11 to 29%. Average improvement in acne scars evaluated by three independent observers was 22% with the 1,320 nm laser compared with 28% with the 1,064 nm laser. The subjects' own grading was 39% with the 1,320 nm laser compared with 37% for the 1,064 nm laser. Prolifometric studies demonstrated comparable improvement, with no statistical difference using either laser. CONCLUSION: These data indicate that both the 1,064 nm laser and the 1,320 nm Nd:YAG laser are safe and effective systems for the nonablative treatment of acne scars, achieving similar improvement. There appears to be a greater response with the 1,064 nm laser system as assessed by the clinical investigators. PMID- 16042935 TI - Effectiveness of photodynamic therapy with topical 5-aminolevulinic acid and intense pulsed light versus intense pulsed light alone in the treatment of acne vulgaris: comparative study. AB - BACKGROUND: Photodynamic therapy (PDT) involves the activation of a photosensitizing agent by light to produce oxygen intermediates that destroy target tissues. Topical 5-aminolevulinic acid (ALA) is converted to protoporphyrin IX, a very potent photosensitizer, which accumulates in human skin, particularly in the epidermis and its appendages. OBJECTIVE: To study the effect of PDT in acne vulgaris using topical ALA and intense pulsed light (IPL). METHODS: Thirteen individuals with varying degrees of acne were treated after a 3 week washout period. Twenty percent ALA hydrochloride (DUSA Pharmaceuticals, Wilmington, MA, USA) was applied to half of the face, and after 3 hours, the whole face was exposed to intense pulsed light (Quantum SR, Lumenis, Santa Clara, CA, USA) using a 560 nm cutoff filter starting at a fluence of 26 J/cm. The procedure was done twice at 2-week intervals, and the patients were clinically evaluated on the second, fourth, and eighth weeks. RESULTS: All patients had no apparent improvement on the second week on both facial halves. In fact, some of the patients developed acute acneiform eruptions on the side treated with ALA. By the fourth week, however, most of the patients had visible improvement of facial acne that was more significant on the ALA-treated side of the face. This persisted until the eighth week post-treatment. On the other hand, the facial half treated with intense pulsed light only showed a return to baseline of their facial acne. CONCLUSION: ALA-IPL are beneficial in the management of acne vulgaris and may be used in combination with other forms of acne treatment or may be an alternative treatment for patients who do not want to take systemic retinoids. PMID- 16042936 TI - Multipass vector (mpave) technique with nonablative radiofrequency to treat facial and neck laxity. AB - BACKGROUND: Redundant facial and neck skin is a major feature of aging and historically has been corrected surgically. Recently, monopolar radiofrequency application has been introduced for nonablative tissue tightening of skin by volumetric heating of the deep dermis. It has been able to improve neck and cheek laxity and periorbital rhytides and to elevate eyebrows. However, questions remain as to the ideal parameters needed to optimize the use of radiofrequency. OBJECTIVE: To determine the safety and report on the efficacy of a radiofrequency application that involves a multipass vector (mpave) technique to target facial and neck skin laxity. METHODS: Twenty-five patients (skin types I to V) with mild to severe facial and neck laxity received one treatment session with monopolar radiofrequency. Treatment parameters, adverse events, and digital photographs were recorded. All patients were treated with a multipass vector technique consisting of four to five passes targeted over areas of skin that would most improve facial laxity. The multipass vector (mpave) treatment approach is described. Energy levels ranged from 62 to 91 J/cm2 per pulse. RESULTS: All patients experienced some immediate erythema and edema, which had completely resolved in most patients within 48 hours. No scarring or dyspigmentation was noted on follow-up at 6 and 12 weeks. Photographic analysis of pre- and post treatment digital images revealed cosmetic improvement in facial and neck laxity in 96% of patients. The majority of patients demonstrated a moderate or better improvement. Stacked pulses in the submental region were shown to reduce fat. CONCLUSIONS: The direct application of monopolar radiofrequency to facial and neck skin using a multipass vector (mpave) treatment approach was safely tolerated in patients of all skin types. Patient satisfaction correlated well with photographic analysis, and the technique was shown to be efficacious for most patients. PMID- 16042937 TI - Measurement of the anatomic position of the corrugator supercilii. AB - OBJECTIVE: To standardize the location of the corrugator supercilii by using known facial landmarks. METHODS: The corrugator supercilii points of origin, insertion, and maximal muscle thickness were measured relative to the nasion and supraorbital notch in 21 cadavers. Comparisons of measurements between male and female cadavers were performed using a two-tailed t-test. RESULTS: The location of the corrugator supercilii was variable but was not significantly different between males and females. From the nasion, the mean point of origin was 4 mm lateral and 6 mm superior, the mean point of insertion was 33 mm lateral and 16 mm superior, and the mean maximal muscle thickness was 16 mm lateral and 10 mm superior. At their point of greatest thickness, most corrugators were found to lie below the upper border of the superciliary arch. CONCLUSIONS: There is variability in the location of the corrugator supercilii that is not gender dependent. When injecting botulinum toxin into the corrugator supercilii, it might be advantageous if the lower forehead skin of the brow is elevated, thus allowing for more precise injection. This may achieve weakness of the targeted muscle with lower doses of botulinum toxin, thus lessening the risk of eyelid and brow ptosis. PMID- 16042938 TI - Micropigmentation: tattooing for medical purposes. AB - BACKGROUND: Micropigmentation, also known widely as tattooing, is being commonly used esthetically to camouflage various medical conditions related to dermatology and plastic surgery. OBJECTIVE: The aim was to review the procedure of tattooing and its various latest medical indications. METHODS: Peer review of the literature on micropigmentation through a MEDLINE search was done to enumerate its various medical indications. RESULTS: The literature review revealed widespread acceptance of micropigmentation for a spectrum of diseases of cosmetic importance, especially in mucosal vitiligo. Micropigmentation is also being used for various medical indications, such as burn scars, alopecia areata, and nipple areola reconstruction. CONCLUSIONS: The procedure is relatively easy, provides permanent camouflage, and is generally devoid of any significant adverse effects. However, a number of infections can be transmitted from one patient to another if the universal precautions for sterilization of instruments used for micropigmentation are not adhered to. PMID- 16042939 TI - Cardiac devices and electromagnetic interference revisited: new radiofrequency technologies and implications for dermatologic surgery. AB - BACKGROUND AND OBJECTIVE: As the number of patients with implantable cardiac devices escalates, inadvertent electromagnetic interference (EMI) by electrosurgery carries increasing concern for patient safety. Given the trend for dermatologic care delivery by nonphysician providers, supervising physicians must carefully consider the adequacy of perioperative evaluation in elective or cosmetic settings. Rapidly evolving technology also calls into question potential EMI of new therapeutic modalities (radiofrequency resurfacing, electrochemotherapy, and endovenous ablation). The most recently published recommendations (1998) for care of dermatologic patients with cardiac devices do not differ significantly from guidelines proposed in 1975. These recommendations, based on complications experienced during noncutaneous surgery, are exceedingly conservative and do not take into consideration the different electrosurgical modalities and varying degrees of EMI risk. In addition, modern cardiac devices have evolved significantly with increased sophistication in protection from EMI. A survey of dermatologic surgeons demonstrated low compliance with these existing guidelines. METHODS, RESULTS, AND CONCLUSIONS: Based on a review of the literature and current electrosurgical and cardiac devices, we discuss the potential impact of conventional dermatologic electrosurgery and new technologies. We propose that new perioperative guidelines are needed to provide appropriate safety, facile implementation, and cost-effective care for patients with modern implantable cardiac devices. PMID- 16042940 TI - Extranasal applications of the bilobed flap. AB - BACKGROUND: The bilobed flap has proven its utility in the reconstruction of distal nasal defects. The mechanics and geometry of the bilobed flap must be clearly understood to achieve these reproducible results. The novelty of the bilobed flap as it is applied to extranasal reconstruction is that the surgeon has some degree of variance in designing the size of the individual lobules of the flap, with a lower risk of secondary tissue distortion. Several variations of the bilobed flap are described, with emphasis on technique and the unique issues involved in its extranasal application. OBJECTIVE: Our experience with the multiple extranasal applications of the bilobed flap is described to demonstrate the usefulness of this reconstructive technique. METHODS: The general technique of the bilobed flap is described, with emphasis on the extranasal variations. Five anatomic locations are presented, with unique considerations and photographs. RESULTS: The bilobed flap as it is applied to extranasal reconstruction allows the surgeon some degree of variance in designing the size of the individual lobules of the flap (compared with the distal nose), with a lower risk of secondary tissue distortion. The flap must be carefully designed to place the incision lines as close to relaxed skin tension lines as possible. A meticulous suture technique is important in camouflaging the complex suture line of this flap, which in no circumstance completely parallels the relaxed skin tension lines. CONCLUSION: We hope to expand the use of this versatile flap to extranasal locations. It can produce the equivalent excellent functional and esthetic results compared with its more common nasal application. PMID- 16042941 TI - Subcutaneous pedicle limberg flap for facial reconstruction. AB - BACKGROUND: A residual dog-ear arising from the traditional Limberg flap transposition sometimes influences the cosmetic outcomes, and the Limberg flap with a subcutaneous pedicle is an innovation applicable to a variety of facial defects and free from the dog-ear. OBJECTIVE: To investigate the outcome of the subcutaneous pedicle Limberg flap for the reconstruction of medium-sized facial skin defects. MATERIALS AND METHODS: From August 2002 to June 2004, the subcutaneous pedicle Limberg flap was designed to repair facial skin defects in 17 patients (19 flaps), and the size of the lesions ranged from 2.0 x 1.9 cm to 5.0 x 4.5 cm. RESULTS: All flaps survived with primary healing postoperatively. The patients were followed up from 1 to 22 months, and functionally and cosmetically satisfactory outcomes were achieved. CONCLUSIONS: This subcutaneous pedicle Limberg flap provides a competitive repair alternative for the treatment of medium-sized skin defects in the face. PMID- 16042942 TI - Porcine sebaceous cyst model: an inexpensive, reproducible skin surgery simulator. AB - BACKGROUND: Surgical simulators are an established part of surgical training and are regularly used as part of the objective structured assessment of technical skills. Specific artificial skin models representing cutaneous pathology are available, although they are expensive when compared with pigskin. The limitations of artificial skin models include their difficulty in representing lifelike cutaneous pathology. OBJECTIVE: Our aim was to devise an inexpensive, reproducible surgical simulator that provides the most lifelike representation of the sebaceous cyst. materials and methods: Pigskin, either pig's feet/trotters or pork belly, was incised, and a paintball was inserted subcutaneously and fixed with cyanoacrylic glue. RESULTS: This model has regularly been used in cutaneous surgical courses that we have organized. Either adding more cyanoacrylic glue or allowing more time for the paint ball to absorb fluid from surrounding tissue can also adjust the degree of difficulty. CONCLUSIONS: The degree of correlation with lifelike cutaneous pathology is such that we recommend that all courses involved in basic skin surgery should consider using the porcine sebaceous cyst model when teaching excision of sebaceous cysts. PMID- 16042943 TI - Repair of a large surgical defect involving the preauricular cheek and sideburn. PMID- 16042944 TI - Central white network: an additional dermoscopic feature for the diagnosis of dermatofibroma. PMID- 16042945 TI - Botulinum toxin a in the treatment of chromhidrosis. AB - BACKGROUND: Chromhidrosis is an uncommon disorder characterized by secretion of colored sweat by apocrine glands, typically localized to the face or axilla. The current treatments available for chromhidrosis are time consuming and frequently ineffective. OBJECTIVE: Our purpose is to demonstrate a novel approach to the treatment of apocrine chromhidrosis. METHODS: We report a case of apocrine chromhidrosis successfully treated with botulinum toxin A (BTX-A; Botox). RESULTS: BTX-A therapy successfully controlled facial chromhidrosis, and the effects were visible at 19 weeks post-treatment. The therapeutic benefits may be attributed to its inhibitory effects on cholinergic stimulation, adrenergic stimulation, and substance P release, although further studies are necessary to elucidate the precise mechanism of action. CONCLUSION: This report demonstrates a new therapeutic approach to patients suffering from chromhidrosis. PMID- 16042946 TI - Postradiation sarcoma: case report and review of the potential complications of therapeutic ionizing radiation. AB - BACKGROUND: Radiation therapy is an acceptable treatment choice for many cutaneous malignancies. A potential late sequela of ionizing radiation is the development of secondary neoplasms within the treatment field. Although there are well-known cutaneous syndromes in which the use of ionizing radiation is contraindicated, in other clinical situations, physicians may not fully realize the risks associated with this therapeutic modality. OBJECTIVE: Through a case report presentation, a potential adverse consequence of radiation therapy will be discussed. A subsequent review of the literature should allow clinicians to better understand the potential risks of therapeutic radiation. METHODS: A case report and review of the literature are provided. RESULTS: A healthy black male with an unusual distribution of lower extremity squamous cell carcinomas in situ developed a malignant fibrous histiocytoma after radiation therapy. CONCLUSION: Postradiation sarcomas are uncommon complications of radiation therapy; however, the significant metastatic capabilities of these tumors demand that clinicians be aware of the potential risks of primary radiation therapy in the treatment of cutaneous tumors. PMID- 16042947 TI - Infiltrative trichilemmal carcinoma treated with 5% imiquimod cream. AB - BACKGROUND: Trichilemmal carcinoma (TLC) is a rare cutaneous neoplasm that occurs in the sun-exposed skin of the elderly. Although the clinical behavior of TLC appears to be relatively indolent, wide excision or Mohs micrographic surgery is the recommended treatment owing to its locally aggressive growth. Recently, imiquimod has become an important part of the armamentarium in the treatment of nonmelanoma skin cancer. OBJECTIVE: This article serves to remind dermatologists that in addition to Mohs micrographic surgery and surgical excision, imiquimod 5% cream may have a role in the treatment of TLC. METHODS: In addition to a review of the literature, we present a case of TLC successfully treated with 5% imiquimod cream. CONCLUSION: Its nonsurgical approach and excellent cosmetic result make imiquimod 5% cream another promising therapeutic option for TLC, even though it is histologically infiltrative. PMID- 16042948 TI - Re: anterior face-lift for correction of middle face aging utilizing a minimally invasive technique. PMID- 16042949 TI - A case of pseudoxanthoma elasticum? PMID- 16042950 TI - Nodular melanoma: controversies and considerations for containment. PMID- 16042951 TI - Comments on postauricular pedicle flaps. PMID- 16042952 TI - Interview with Nancy Sung, Kate Ahlport, and Enriqueta C. Bond, Burroughs Wellcome Fund. PMID- 16042956 TI - Developing the physician-scientist, Part 5: Interview with James R. Gavin III, MD, PhD. PMID- 16042957 TI - Alcohol abuse and acute lung injury: epidemiology and pathophysiology of a recently recognized association. AB - Alcohol is the most commonly used and abused drug in the United States. The deleterious health effects of alcohol can be attributed both to its acute intoxicating effects, which result in temporary impairment of judgment and motor skills, and to its more chronic and toxic effects on the liver, pancreas, heart, and brain, all of which may result in irreversible organ damage. Although recognized for more than a century as a major risk factor for pneumonia, alcohol abuse was until recently perceived to have no significant effects on lung structure and/or function. However, within the past decade, epidemiologic studies have revealed that alcohol abuse independently increases the risk of acute respiratory distress syndrome (ARDS) two- to fourfold in patients with sepsis or trauma and may play a role in ARDS pathogenesis in as many as half of all patients with the syndrome. Although alcohol abuse alone does not cause acute lung injury, it renders the lung susceptible to dysfunction in response to the inflammatory stresses of sepsis, trauma, and other clinical conditions recognized to cause ARDS. Recent investigations in both animal models of chronic ethanol ingestion and in human subjects with a history of alcohol abuse have explored this previously unrecognized connection between alcohol and acute lung injury and have uncovered multiple derangements, which we now characterize as the "alcoholic lung." This review summarizes the epidemiologic association between alcohol abuse and acute lung injury and the recent experimental findings that are unraveling the underlying pathophysiology. PMID- 16042958 TI - Hemoglobin serves to protect Plasmodium parasites from nitric oxide and reactive oxygen species. AB - Our understanding of how the host immune response kills Plasmodium, the causative agent of malaria, is limited and controversial. One widely held belief is that reactive oxygen species are crucial for controlling parasite replication. One of the hallmarks of blood-stage malaria is the cyclic rupture of erythrocytes by the parasite, which releases free hemoglobin into the circulation. We propose that this free hemoglobin, as well as the hemoglobin within the erythrocyte and surrounding the parasite, effectively shields Plasmodium from reactive oxygen species well in excess of those achievable in vivo. PMID- 16042959 TI - Impaired lung vascular endothelial growth factor in extremely premature baboons developing bronchopulmonary dysplasia/chronic lung disease. AB - BACKGROUND: Preterm infants exposed to O2 with mechanical ventilation often develop bronchopulmonary dysplasia (BPD), a form of chronic lung disease (CLD). The pathogenesis of BPD/CLD involves dysmorphic microvasculature and disrupted alveolarization. This may be due to impaired vascular endothelial growth factor (VEGF) and VEGF receptor expression. METHODS: To examine the ontogeny of VEGF and VEGF receptors in baboon lungs from 125 to 185 (term) days gestation and to determine whether exposure to O2 and mechanical ventilation alter these ontogenic profiles, we examined lung specimens from three O2-exposed groups: (1) animals delivered at 125 days gestation and exposed to O2 for 14 days as needed; (2) animals delivered at 140 days gestation and exposed to O2 for 10 days as needed; and (3) animals delivered at 140 days gestation and exposed to 100% O2 for 10 days. Lungs from gestational age-matched controls were also examined at 125, 140, 160, 175, and 185 (term) days. RESULTS: VEGF189 was the most abundant splice variant in the lungs at all stages of development. Extremely premature baboons developing BPD/CLD had higher lung VEGF121 messenger ribonucleic acid (mRNA) expression. However, transcripts for VEGF189, VEGF165, and VEGF receptors (Fms like tyrosine kinase-1 [Flt-1], kinase-insert domain receptor [KDR]/fetal liver kinase-1 [Flk-1], and neuropilin 1) were suppressed in the BPD models. CONCLUSIONS: We conclude that impaired VEGF and VEGF receptor mRNA expression in lungs from extremely premature baboons developing BPD/CLD may contribute to dysmorphic microvasculature and disrupted alveolarization. PMID- 16042960 TI - Patient knowledge improves glycemic control: is it time to go back to the classroom? AB - BACKGROUND: Over the last two decades, pharmaceutical intervention for the treatment of type 2 diabetes has expanded. Studies over this same time demonstrated the benefits of tight glycemic control. Unfortunately, despite the availability of novel therapies, glycemic control remains problematic. Nonpharmacologic interventions need to be explored, including patient empowerment. Improving patient knowledge of diabetes may ultimately improve glycemic control. To test this hypothesis, we compared patients' diabetes knowledge with their glycemic control. METHODS: The Michigan Diabetes Knowledge Test, designed by the University of Michigan, was administered to patients with type 2 diabetes at three University of New Mexico primary care clinics. Patient records were reviewed. The most recent hemoglobin A1c (HbA1c) value was recorded. The data were analyzed using linear regression analysis. RESULTS: Seventy-seven patients completed surveys and had HbA1c values available. Only questions 1 to 14 of the 23-question survey were used because they pertained specifically to type 2 diabetes. HbA1c was inversely correlated with the number of questions answered correctly on the test (r = -.337, p < .003). Using "all subsets" regression, a correct response to questions 1, 3, and 9 specifically correlated with a lower HbA1c (p < .0001). CONCLUSIONS: These results demonstrate that an inverse linear relationship exists between performance on this diabetes test and HbA1c values. Improvement in patient knowledge of diabetes and the importance of treatment may indeed improve glycemic control and ultimately decrease complications. Studies aimed at empowering patients with disease knowledge may help control the ramifications of the growing diabetes epidemic. PMID- 16042961 TI - Effect of diclofenac on experimental pleurodesis induced by tetracycline in rabbits. AB - BACKGROUND AND OBJECTIVE: Pleurodesis is a frequently preferred procedure in thoracic surgery, and many factors may affect the process. We aimed to determine whether the administration of systemic diclofenac sodium diminishes the effectiveness of the pleurodesis induced by intrapleural tetracycline in rabbits. METHODS: Twelve male New Zealand rabbits that received tetracycline 35 mg/kg intrapleurally were allocated into two groups. The first group (diclofenac group, n = 6) received 2 mg/kg diclofenac sodium intramuscularly for 10 days, and the second group (control group, n = 6) received acetaminophen 30 mg/kg orally for 10 days after the pleurodesis procedure. The rabbits were sacrificed after 28 days, and the pleural spaces were assessed grossly for evidence of pleurodesis and microscopically for evidence of fibrosis, inflammation, and collagenization. RESULTS: The mean macroscopic pleurodesis score of the diclofenac group was 2.16 +/- 0.40 compared with 2.83 +/- 0.40 in the control group (p = .027). The mean microscopic pleurodesis score of the diclofenac group was 2. 3 +/- 1.03, whereas it was 3.5 +/- 0.54 in the control group (p = .045). CONCLUSION: The administration of diclofenac sodium for 10 days following tetracycline pleurodesis reduces the effectiveness of pleurodesis in rabbits. PMID- 16042963 TI - Monitoring the organization and dynamics of bovine hippocampal membranes utilizing Laurdan generalized polarization. AB - Organization and dynamics of cellular membranes in the nervous system are crucial for the function of neuronal membrane receptors. The lipid composition of neuronal cells is unique and has been correlated with the increased complexity in the organization of the nervous system during evolution. Previous work from our laboratory has established bovine hippocampal membranes as a convenient natural source for studying neuronal receptors such as the G-protein coupled serotonin1A receptor. In this paper, we have explored the organization and dynamics of bovine hippocampal membranes using the amphiphilic environment-sensitive fluorescent probe Laurdan. Our results show that the emission spectra of Laurdan display an additional red shifted peak as a function of increasing temperature in native as well as cholesterol-depleted membranes and liposomes made from lipid extracts of the native membrane. Interestingly, wavelength dependence of Laurdan generalized polarization (GP) in native membranes indicates the presence of an ordered gel like phase at low temperatures, whereas characteristics of the liquid-ordered phase are observed at high temperatures. Similar experiments performed using cholesterol-depleted membranes show fluidization of the membrane with increasing cholesterol depletion. In addition, results from fluorescence polarization of DPH indicate that the hippocampal membrane is fairly ordered even at physiological temperature. The temperature dependence of Laurdan excitation GP provides a measure of the apparent thermal transition temperature and extent of cooperativity in these membranes. Analysis of time-resolved fluorescence measurements of Laurdan shows reduction in mean fluorescence lifetime with increasing temperature due to change in environmental polarity. These results constitute novel information on the dynamics of hippocampal membranes and its modulation by cholesterol depletion monitored using Laurdan fluorescence. PMID- 16042964 TI - Effects of galanin-like peptide (GALP) on locomotion, reproduction, and body weight in female and male mice. AB - Galanin-like peptide (GALP) has been implicated in the neuroendocrine regulation of both feeding and reproduction. In male rodents and primates, intracerebroventricular (icv) infusions of GALP stimulate luteinizing hormone (LH) release, induce Fos expression in brain areas implicated in feeding and reproduction, and affect food intake and body weight in rodents. In gonad-intact and castrated male rats, icv administration of GALP also stimulates male sexual behavior. While the effects of GALP on male physiology and behavior are well documented, no studies have addressed such a role of GALP in females. We tested the effects of icv GALP infusions on LH release, locomotor activity, motor control, and body weight regulation in adult ovariectomized female mice hormonally primed with estradiol benzoate and progesterone. In addition, sexually experienced male and female mice were treated with GALP and tested for sexual behavior. In females, GALP reduced open-field locomotor activity, the ability to maintain grip on an accelerating rotarod, and 24-h body weight in a dose dependent manner. GALP also increased LH secretion in female mice, an effect that was blocked by pre-treatment with Antide, a gonadotropin-releasing hormone (GnRH) type-1 receptor antagonist. GALP infusions slightly decreased the occurrence of lordosis behavior in female mice and significantly increased the latencies with which females displayed receptivity. Unlike previous reports in male rats, GALP inhibited male sexual behavior in mice. Our data indicate that in female mice, GALP stimulates LH release via GnRH, and decreases body weight, motor control, and locomotor activity via GnRH-independent pathways. Furthermore, our sexual behavior and locomotor findings suggest species-specific differences in the mechanism and/or location of GALP action in the brains of rats and mice. PMID- 16042965 TI - Behavioral specificity of non-genomic glucocorticoid effects in rats: effects on risk assessment in the elevated plus-maze and the open-field. AB - The rapid effects of glucocorticoids on various behaviors suggest that these hormones play a role in rapidly coping with challenging situations. The variety of behaviors affected in different situations raise, however, questions regarding the specificity and roles of glucocorticoids in controlling behavior. To clarify this issue, we assessed the rapid behavioral effects of glucocorticoids in the elevated plus-maze (EPM) and the open-field (OF) tests in male rats. Both tests measure three different kinds of behavioral responses: locomotion, anxiety-like behaviors (central area and open arm exploration in the OF and EPM tests, respectively), and risk assessment (investigating aversive areas in a stretched attend posture). The acute inhibition of glucocorticoid synthesis by metyrapone decreased risk assessment but did not affect locomotion and anxiety-like behaviors. Corticosterone administration increased risk assessment, without affecting locomotion and anxiety-like behaviors. Moreover, plasma corticosterone levels measured immediately after testing strongly correlated with the intensity of risk assessment. The effects of corticosterone were rapid, as occurred even when the hormone was injected 2 min before behavioral testing. In addition, the effect was resistant to protein synthesis inhibition. These data demonstrate that glucocorticoids are able to increase specifically risk assessment behaviors by non-genomic mechanisms in two different, novelty-related, non-social challenging situations. Thus, glucocorticoids appear to rapidly induce specific behavioral adjustments to meet immediate requirements set by the challenge. These data support earlier assumptions on the role of glucocorticoids in coping, and it can be hypothesized that the rapid activation of the HPA-axis may play a role in forming coping responses. PMID- 16042966 TI - The social context of imitation in infancy. AB - Infants increasingly generalize deferred imitation across environmental contexts between 6 and 18 months of age. In three experiments with 126 6-, 9-, 12-, 15-, and 18-month-olds, we examined the role of the social context in deferred imitation. One experimenter demonstrated target actions on a hand puppet, and a second experimenter tested imitation 24h later. When the second experimenter was novel, infants did not exhibit deferred imitation at any age; when infants were preexposed to the second experimenter, all of them did. Imitating immediately after the demonstration also facilitated deferred imitation in a novel social context at all ages but 6 months. Infants' pervasive failure to exhibit deferred imitation in a novel social context may reflect evolutionary selection pressures that favored conservative behavior in social animals. PMID- 16042967 TI - Selective attention and cyclopean motion processing. AB - The effect of diverted selective attention on the induction of the cyclopean motion aftereffect (aftereffect induced from dynamic disparity information) was investigated. The luminance motion aftereffect was examined for comparison. During diverted-attention trials, observers ignored background adapting motion and performed a low-load or high-load rapid serial visual presentation (RSVP) task presented in the center of the motion display. Baseline motion aftereffects were obtained with no diverted attention. The results showed that the cyclopean motion aftereffect, similar to the luminance motion aftereffect, declined only modestly under diverted-attention conditions. Selective attention appears to play a modest role in the visual processing of cyclopean motion. PMID- 16042968 TI - Central and near peripheral retinal contributions to the depth-of-focus using naturalistic stimulation. AB - Although the depth-of-focus (DOF) has been investigated separately in the central retina and in the near retinal periphery, knowledge about their combined relative contribution to overall blur perception has remained unknown. In the present study, the DOF was measured psychophysically with a naturalistic pictorial stimulus as a function of spatial extent across the near retinal periphery under monocular Badal viewing conditions with accommodation paralyzed. The group mean total DOF progressively increased linearly with target size. Based on the individual DOF responses, the group was categorized into two subgroups: a predominantly centrally-driven and a centrally plus peripherally-driven subgroup. The results implicated partial cone pooling of blur information, as well as influence from perceptual, attentional, and optical aspects. However, the subgroup response profiles suggested individual differences in the weighting of the near peripheral blur information at the retinal level, and perhaps at higher level areas of the visual system, involving spatial integration and global attentional processing. PMID- 16042969 TI - Idiosyncratic initiation of saccadic face exploration in humans. AB - Visual processing and subsequent action are limited by the effectiveness of eye movement control: where the eyes fixate determines what part of the visual environment is seen in detail. Visual exploration consists of stereotypical sequences of saccadic eye movements which are known to depend upon both external factors, such as visual stimulus features, and internal cognition-related factors, such as attention and memory. However, how these two factors are balanced is unknown. One determinant might be the familiarity or ecological importance of the visual stimulus being explored. Recordings of saccades for human face stimuli revealed that their exploration was subject to strong individual biases for the initial saccade direction: subjects tended to look first to one particular side. We attribute this to internal factors. In contrast, exploration of landscapes, fractals or inverted faces showed no significant direction bias for initial saccades, suggesting more externally driven exploration patterns. Thus the balance between external and internal factors in scene exploration depends on stimulus type. An analysis of saccade latencies suggested that this individual preference for first saccade direction during face exploration leads to higher effectiveness through automation. The findings have implications for the understanding of both normal and abnormal eye movements. PMID- 16042970 TI - Proceedings of the International Symposium on Peptide Receptors, Montreal, Canada, 2004. PMID- 16042971 TI - Kallikrein-kinin system in hepatic experimental models. AB - The purpose of this brief review is to describe some characteristics of the kallikrein-kinin system (KKS) in the liver. The liver synthesizes kininogens and prekallikrein and the synthesis of both proteins is increased in rats during the acute phase reaction. It is also the main organ to clear tissue as well as plasma kallikrein from the circulation in normal and pathological conditions. Bradykinin (BK), yielded by the kallikrein-kinin system, is a potent arterial hypotensive peptide, but in the liver it induces a portal hypertensive response. The portal hypertensive action of bradykinin is mediated by B2 receptors located on sinusoidal cells of the periportal region and is followed by its hydrolysis by angiotensin-converting enzyme, which is primarily present in the perivenous (centrolobular) region. PMID- 16042972 TI - Bradykinin B2 type receptor activation regulates fluid and electrolyte transport in the rabbit kidney. AB - Bradykinin is an important autacoid produced in the kidney, regulating both renal function and blood pressure. In vivo studies in anesthetized rabbits, revealed that BK induced diuresis (UV), natriuresis (U(Na)V) and was not associated with renal hemodynamic changes. These diuretic and natriuretic effects were blocked by the BK-B2 antagonist HOE-140. BK also inhibits vasopressin (AVP)-stimulated water flow (L(p)) in microperfused rabbit cortical collecting ducts (rCCD), in a concentration-dependent fashion, consistent with its in vivo diuretic effects. BK B1 antagonist Leu8-des-Arg9-BK did not alter the effect of BK on Lp, but HOE-140 completely blocked the inhibitory effects of BK on Lp. While BK did not increase [Ca2+]i in fura-2 loaded freshly microdissected rCCD, BK increased [Ca2+]i in immortalized cultured rCCD cells demonstrating different signaling mechanisms are activated by BK in microdissected versus cultured rCCD. In microperfused rCCD, neither the protein kinase C inhibitor staurosporine nor the phospholipase C (PLC) inhibitor U-73,122 attenuated the BK response arguing against activation of PLC/PKC by BK in rCCD. We conclude: (1) BK induces UV and U(Na)V by a BK-B(2) receptor; (2) BK inhibits AVP-stimulated Lp by a BK-B2 receptor suggesting that its effects on Lp are not via a PLC/PKC; (3) finally, BK raises [Ca2+]i in rCCD cells by a BK-B2 receptor mechanism. PMID- 16042973 TI - Hypotensive effects of hemopressin and bradykinin in rabbits, rats and mice. A comparative study. AB - Hemopressin is a novel vasoactive nonapeptide derived from hemoglobin's alpha chain as recently reported by Rioli et al. [Rioli V, Gozzo FC, Heimann AS, Linardi A, Krieger JE, Shida CS, et al. Novel natural peptide substrates for endopeptidase 24.15, neurolysin, and angiotensin-converting enzyme. J Biol Chem 2003;278(10):8547-55]. In anesthetized male Wistar rats, this peptide exhibited hypotensive actions similar to those of bradykinin (BK) when administered intravenously (i.v.), and was found to be metabolized both in vitro and in vivo by several peptidases, including the angiotensin-converting enzyme (ACE). In this study, these findings were expanded upon by examining: (i) the degradation kinetics following incubation with ACE purified from rabbit lung and (ii) the blood pressure lowering effects of HP and BK injected i.v. or intra-arterially (i.a.) in male rabbits, rats, and mice. Our findings demonstrate that, in vitro, HP and BK are both degraded by ACE, but at different velocity rates. Furthermore, both HP and BK induced transient hypotension in all animals tested, although the responses to HP relative to the administration sites were significantly lower (by 10-100-fold) on an equimolar basis compared to those of BK. In rabbits, the decrease of blood pressure induced by HP (10-100 nmol/kg) did not differ whether it was administered i.v. or i.a., suggesting an absence of pulmonary/cardiac inactivation in contrast to BK (0.1-1 nmol/kg). The in vivo effect of HP was significantly potentiated in rabbits immunostimulated with bacterial lipopolysaccharide (LPS), but was unaffected by both the B2 receptor antagonist HOE 140 (0.1 micromol/kg) and captopril (100 microg/kg), contrary to BK. Therefore, HP acts as a weak hypotensive mediator, which does not activate kinin B2 receptors, but uses a functional site and/or signaling paths appearing to be up-regulated by LPS. PMID- 16042974 TI - Increases of spinal kinin receptor binding sites in two rat models of insulin resistance. AB - An autoradiographic study was conducted to determine whether kinin receptors are altered in the rat spinal cord in two experimental models of chronic hyperglycemia and insulin resistance. Sprague-Dawley rats were given 10% d glucose in their drinking water alone or with insulin (9 mU/kg/min with osmotic pumps) for 4 weeks. Both groups and control rats were treated either with a normal chow diet or with an alpha-lipoic acid-supplemented diet as antioxidant therapy. After 4 weeks of treatment, glycemia, insulinemia, blood pressure, insulin resistance index, the production of superoxide anion in the aorta and the density of B2 receptor binding sites in the dorsal horn were significantly increased in the two models. These effects were prevented or attenuated by alpha lipoic acid. In contrast, B2 receptor binding sites of most spinal cord laminae were increased in the glucose group only and were not affected by alpha-lipoic acid. Results show that chronic hyperglycemia associated with insulin resistance increases B1 and B2 receptor binding sites in the rat spinal cord through distinct mechanisms, including the oxidative stress for the B1 receptor. PMID- 16042975 TI - The spinal basis of opioid tolerance and physical dependence: Involvement of calcitonin gene-related peptide, substance P, and arachidonic acid-derived metabolites. AB - Chronic opioid use in the management of pain is limited by development of analgesic tolerance and physical dependence. The mechanisms underlying tolerance dependence are not entirely clear, however, recent evidence suggests that spinal adaptations leading to increased activity of sensory neuropeptides (calcitonin gene-related peptide (CGRP), substance P) and their downstream signaling messengers derived from metabolism of arachidonic acid: prostaglandins (PG), lipoxygenase (LOX) metabolites, and endocannabinoids, plays an important role in this phenomenon. In this communication we review the evidence implicating these factors in the induction and expression of opioid tolerance and physical dependence at the spinal level. PMID- 16042976 TI - New challenges in the study of the mammalian tachykinins. AB - There is an expanding repertoire of mammalian tachykinins produced by a variety of tachykinin genes, gene splicing events and peptide processing. Novel tachykinin-binding molecules/receptors are proposed, but only, three tachykinin receptors are identified with certainty. The question remains - do more tachykinin receptors exist or is there just the need to reappraise our understanding of the known receptors? The tachykinin NK1 receptor, the preferred receptor for both substance P and the peripheral SP-like endokinins, exists in several tissue-specific conformations and isoforms and may provide some clues. This review addresses recent advances in this exciting field and raises challenging new concepts. PMID- 16042977 TI - Tachykinin peptides and receptors: putting amphibians into perspective. AB - The tachykinins form one of the largest peptide families in nature. In this review, we describe the comparative features of the tachykinin peptides and their receptors, focusing particularly on amphibians. We also summarize our systematic studies of the localization, characteristics, and actions of bufokinin, a toad substance P-related peptide, in its species of origin. In addition, we discuss the establishment of multiple isoforms of the NK1-like receptor in the toad, and their structure, pharmacology and tissue distributions. We conclude that tachykinin peptides and receptors are well conserved in terms of their structures, physiological functions and coupling mechanisms during tetrapod evolution. PMID- 16042978 TI - Effect of neurokinin-1 receptor antagonists on serotoninergic, noradrenergic and hippocampal neurons: comparison with antidepressant drugs. AB - Neurokinin-1 (NK1) receptor antagonists have been reported to possess antidepressant and anxiolytic properties in controlled trials. Since antidepressant and anxiolytic drugs act mainly by enhancing serotonin (5-HT) and norepinephrine (NE) neurotransmission in forebrain areas, the main focus of the present review is to critically examine the electrophysiological effects of NK1 receptor antagonists on serotoninergic and noradrenergic neurons, and then hippocampal neurons. It is concluded that NK1 antagonists increase the firing and burst activity of 5-HT neurons, increase burst activity of NE neurons, and modulate postsynaptic transmission at the hippocampus level. Further research is needed in order to develop more selective ligands for the human NK1 receptor and to gain better knowledge of required brain penetration and optimal pharmacodynamic conditions for their use in patients. PMID- 16042979 TI - Role of serotonin (5-HT) in the antidepressant-like properties of neuropeptide Y (NPY) in the mouse forced swim test. AB - Neuropeptide Y (NPY) is thought to be implicated in depressive disorders. The mouse forced swim test (FST) is an animal model widely used as a predictor of the efficacy of antidepressant drugs. The present study was undertaken to explore the possible contribution of endogenous serotonin (5-HT) systems in the behavioral effects elicited by NPY in this model. The selective serotonin re-uptake inhibitor (SSRI), fluoxetine, was also tested for comparison. 5-HT was depleted prior to testing by the administration of the tryptophan hydroxylase inhibitor p chlorophenylalanine (PCPA; 300 mg/kg, i.p., each day for 3 days; control mice received saline-vehicle over the same period). On the fourth day, mice received NPY (3 nmol, I.C.V.), fluoxetine (16 mg/kg, i.p.) or saline injections before testing in the FST. Both NPY and fluoxetine significantly reduced immobility time in saline-treated control animals. Pre-treatment with PCPA significantly blocked the effects of fluoxetine in the FST, confirming the role of endogenous 5-HT. Similarly, pre-treatment with PCPA also significantly attenuated the anti immobility effects of NPY, thus suggesting a role for 5-HT in the effects of NPY in the FST. Quantitative receptor autoradiography revealed increases in specific [125I][Leu31, Pro34]PYY sites that were sensitive to BIBP3226 (Y1-like sites) in various brain regions. Specific [125I]GR231118 and [125I]PYY(3-36) binding levels were not changed following PCPA treatment, suggesting that depletion of endogenous 5-HT resulted in an apparent increase in the level of Y1 sites in their high-affinity state. Taken together, these results suggest a role for 5-HT related systems in the antidepressant-like properties of NPY. PMID- 16042980 TI - The AT2 receptor--a matter of love and hate. AB - In 1989, the development of specific angiotensin receptor antagonists which distinguish between two angiotensin receptor subtypes (AT1 and AT2) led to a breakthrough in angiotensin research. It turned out, that the AT1 receptor was almost entirely responsible for the "classical" actions of angiotensin II related to the regulation of blood pressure as well as volume and electrolyte balance. However, actions and signal transduction mechanisms coupled to the AT2 receptor remained enigmatic for a long time. The present review summarizes the current knowledge of AT2 receptor distribution, signaling and function with an emphasis on growth/anti-growth, differentiation and the regeneration of neuronal tissue. PMID- 16042981 TI - Angiotensin II-induced increase of T-type Ca2+ current and decrease of L-type Ca2+ current in heart cells. AB - The effect of angiotensin II (Ang II) on the T- and L-type calcium currents (I(Ca)) in single ventricular heart cells of 18-week-old fetal human and 10-day old chick embryos was studied using the whole-cell voltage clamp technique. Our results showed that in both, human and chick cardiomyocytes, Ang II (10(-7)M) increased the T-type calcium current and decreased the L-type I(Ca). The effect of Ang II on both types of currents was blocked by the AT1 peptidic antagonist, [Sar1, Ala8] Ang II (2 x 10(-7)M). Protein kinase C activator, phorbol 12,13 dibutyrate, mimicked the effect of Ang II on the T- and L-type calcium currents. These results demonstrate that in fetal human and chick embryo cardiomyocytes Ang II affects the T- and L-type Ca2+ currents differently, and this effect seems to be mediated by the PKC pathway. PMID- 16042982 TI - The distribution and density of ET-1 and its receptors are different in human right and left ventricular endocardial endothelial cells. AB - Evidence suggests that endocardial endothelial cells (EECs) may play a role in the regulation of cardiac function by releasing ET-1. Furthermore, reports in the literature suggested that differences may exist in peptide receptor distribution between the left and right EECs. In this study, we verified if the distribution and density of ET-1 and its receptors could be different in right as compared to left ventricular EECs, and whether this difference may affect ET-1-induced increase of intracellular calcium. Using immunofluorescence and 3D confocal microscopy, our results showed that in both cell types, the ET(A) receptor is present and is homogeneously distributed throughout the two cell types. The relative density of the ET(A) receptor is similar in both right and left ventricular EECs. The ET(B) receptor is also present in right and left ventricular EECs, however, the relative density of the ET(B) receptor is higher in the nucleus as compared to the cytosol. In addition, the ET(B) receptor density was found to be higher in left EECs as compared to right EECs. In addition, our results showed that ET-1 is present in the cytosol and the nucleus of both types of cells and that the relative density of ET-1 is higher in right as compared to left ventricular EECs. Moreover, using the Fura-2 calcium measurement technique, our results showed that in left ventricular EECs, both ET(A) and ET(B) receptor activation mediated the effect of ET-1 on intracellular calcium, whereas in right ventricular EECs, this effect was solely mediated by the ET(A) receptor. In conclusion, our results showed that ET-1 and its receptors are present in both right and left ventricular EECs. However, the distribution and relative density of ET-1 and its receptors seem to be different in right EECs as compared to left EECs. PMID- 16042983 TI - PTHrP fragments 1-16 and 1-23 do not bind to either the ETA or the ETB endothelin receptors. AB - Because of some isofunctional similarities with endothelin-1 (ET-1), it has been suggested that PTHrP(1-16) and PTHrP(1-23) could interact with osteoblast cells via ETA receptors. To document this interaction, we used the thoracic rat aorta and the guinea-pig lung parenchyma paradigms as ETA and ETB models, respectively. In addition, we also performed a series of competition experiments against [125I]ET-1, using transfected cells expressing the ETA or ETB receptor. So far, no agonistic nor antagonistic activities were observed in the ETA and ETB bioassays with the PTHrP fragments. Furthermore, both fragments were unable to displace [125I]ET-1 bound to cells expressing the ETA or ETB receptor. PMID- 16042984 TI - Development of an efficient strategy for the synthesis of the ETB receptor antagonist BQ-788 and some related analogues. AB - BQ-788 [N-cis-2,6-dimethylpiperidine-1-carbonyl-L-gamma-methylleucyl-D-1 methoxycarbonyltryptophanyl-D-norleucine sodium salt] is a very potent and selective ETB receptor antagonist. The formation of the highly hindered trisubstituted urea functionality in the peptide chain and the carbamination on the indole nitrogen of the tryptophan side chain are major challenges in the synthesis of this particular antagonist. Furthermore, the high cost of the unnatural amino acids in the sequence of BQ-788 and its reported synthesis render this pseudopeptide very expensive to produce. In order to improve the yield and to reduce the number of steps compared to previous reported syntheses, we developed an efficient strategy involving a novel one-pot procedure for the synthesis of a highly hindered trisubstituted urea. Under very mild conditions, the urea was obtained by using triphosgene and sodium iodide. This strategy allowed us to synthesize BQ-788 in seven steps with an overall yield of 53%. We also generalized the use of this powerful methodology by creating some new structural analogues of the cis-2,6-dimethylpiperidine moiety by replacing it with other bulky secondary amines. We evaluated the antagonist properties of those three new analogues of BQ-788 in two bioassays in vitro. These new antagonists were less potent than BQ-788 in an ETB rich preparation and inactive in an ETA rich preparation. PMID- 16042985 TI - Endothelin-1 contributes to the sexual differences in renal damage in DOCA-salt rats. AB - We investigated whether gender differences in renal damage in DOCA-salt hypertension are associated with effects of ovarian hormones and/or endothelin-1 (ET-1). Renal injuries and renal pre-pro-ET-1 mRNA expression were enhanced in male and female ovariectomized (OVX) DOCA rats versus female DOCA rats. Treatment with estrogen plus progesterone or progesterone, but not estrogen alone, attenuated renal damage and pre-pro-ET-1 mRNA expression in OVX DOCA rats. The ETA antagonist BMS182874 greatly ameliorated renal damage in male and OVX DOCA rats. In conclusion, the ovarian hormones have a protective role on the renal structural alterations in female DOCA rats by modulating effects of ET-1, via ETA receptors. PMID- 16042986 TI - Low-affinity state beta1-adrenoceptor-induced vasodilation in SHR. AB - Low-affinity state beta1-adrenoceptor (beta1-AR) was functionally expressed in some blood vessels and was different from beta1, beta2 and beta3-AR. In rat aorta, low-affinity state beta1-AR activation produced an endothelium-independent relaxation which was impaired in spontaneously hypertensive rats (SHRs). In the present work, we investigated whether renin-angiotensin system was involved in this alteration by evaluating the effects of enalapril, an angiotensin converting enzyme (ACE) inhibitor or losartan, an AT1 angiotensin receptor antagonist. Cumulative concentration-response curves to low-affinity state beta1-AR agonists (CGP 12177, cyanopindolol or alprenolol) and to NS 1619, a large conductance Ca2+ activated K+ channels (BK) agonist were performed in denuded aortic rings isolated from control or treated Wistar Kyoto (WKY) rats or SHRs in different experimental conditions. The low-affinity state beta1-AR-mediated aortic vasodilation was impaired in 5 and 12 weeks old SHRs when compared to age-matched WKY. Twelve days enalapril (5 mg/kg/day) or losartan (15 mg/kg/day) treatments reduced systolic blood pressure (SBP) only in 12 weeks old SHRs whereas no significant change was observed in other groups. These treatments improved low affinity state beta1-AR effect only in SHRs groups. In 12 weeks old WKY rats, CGP 12177-induced relaxation was insensitive to glibenclamide, a K(ATP)+ channel blocker, but was reduced by TEA or iberiotoxin, two large conductance Ca2+ activated K+ channel (BK) blockers. The impairment of NS 1619-induced vasodilation in both 5 and 12 weeks old SHRs was restored by enalapril or losartan. These results suggested that improvement of the low-affinity state beta1-AR-mediated vasodilation in 5 and 12 weeks old SHRs could be attributed to enhanced BK channels-induced hyperpolarization in SHRs independently of lowering of SBP. PMID- 16042987 TI - Urotensin II-induced hypotensive responses in Wistar-Kyoto (Wky) and spontaneously hypertensive (Shr) rats. AB - Human urotensin II (hU-II) is a potent vasoactive peptide which modulates some of the functions of the cardiovascular and other systems. The in vivo mechanism of action by which hU-II may influence blood pressure in developmental and pathological conditions, is poorly understood. Herein, the blood pressure effects of hU-II (0.1-10 nmol/kg) injected intravenously (i.v.) were studied on ketamine/xylazine anesthetized male WKY and SHR rats aged 4 and 8 weeks. hU-II elicited dose-dependent decreases in mean arterial pressure in both strains of animals. The hypotensive responses to hU-II were, however, significantly higher in SHR rats, independently of age. Four-week-old SHR rats (which are normotensive) were, however, less responsive than their hypertensive 8-week-old counterparts. A series of pharmacological inhibitors were used to identify putative endogenous (endothelial) factors that might account for the hU-II mediated hypotension in 8-week-old SHR. These include the non-selective nitric oxide synthase inhibitor L-NAME (5 micromol/kg), the non-selective cyclooxygenase inhibitor meclofenamate (16 micromol/kg), the voltage-sensitive and ATP-sensitive K+-channel inhibitors, 4-aminopyridine (5 micromol/kg) and glybenclamide (10 micromol/kg), the cytochrome P450 CYP2C9 inhibitor sulfaphenazole (15 micromol/kg), the cytoskeletal fixation agent phalloidin (15 micromol/kg), the endothelin ETB receptor antagonist BQ-788(35 micromol/kg), the bradykinin B2 receptor antagonist HOE 140 (0.5 micromol/kg), the angiotensin AT2 antagonist PD 123319(10 micromol/kg) and the UT receptor antagonist urantide (10 micromol/kg). These agents were administered i.v. either at 2.5, 10 or 40 min prior hU-II injection (10 nmol/kg). Among these inhibitors, sulfaphenazole and phalloidin were able to reduce hU-II-induced hypotension. This suggests that the vasodepressor effect of hU-II is mediated by UT receptors and relies in part on the release of epoxide related products; increased microvascular permeability may also contribute to the blood pressure lowering effect of hU-II. Since urantide blocks the constrictor effects of hU-II on isolated aorta, but is inactive against the hypotensive action of hU-II in vivo, the results presented in this paper provide, for the first time, evidence for the existence of two different functional sites for hU-II. PMID- 16042988 TI - Development and pharmacological characterization of "caged" urotensin II analogs. AB - Urotensin-II (U-II) is a cyclic 11-amino acid peptide known as a potent mammalian vasoconstrictor. To study some purported intracellular actions of U-II, masked analogs of this peptide, becoming biologically active only upon UV exposure, were developed. Those analogs described as "caged" were derivatized with a photolabile 4,5-dimethoxynitrobenzyl group on the side chain of Lys-8 or Tyr-9. Both caged analogs of U-II showed a major decrease in their affinity towards the UT receptor. Nevertheless, upon UV irradiation, the native and biologically active U II peptide was recovered. Thus, this work describes the development of new "caged" U-II derivatives and demonstrates that vasoactivity of U-II can be controlled by masking and unmasking two key residues. PMID- 16042989 TI - Nociceptin/orphanin FQ inhibits electrically induced contractions of the human bronchus via NOP receptor activation. AB - Nociceptin/orphanin FQ (N/OFQ) has been reported to inhibit neurogenic contractions in various tissues, including guinea pig airways. In the present study, we investigated the ability of N/OFQ to affect cholinergic contractions of human bronchi elicited by electrical field stimulation (EFS). Tissues were obtained from 23 patients undergoing surgery for lung cancer. EFS (20 Hz, 320 mA, 1.5 ms, 10 s) was applied five times every 20 min. Contractions induced by EFS were abolished by either TTX (1 microM) or atropine (1 microM) and concentration dependently (10 nM-1 microM) inhibited by N/OFQ (Emax, 11.5+/-1.8% inhibition). The inhibitory effects of N/OFQ were mimicked by the N/OFQ receptor (NOP) ligand [Arg14, Lys15]N/OFQ which displayed however, higher significant maximal effects (17.7+/-2.9% inhibition, P<0.05). The actions of N/OFQ and [Arg14, Lys15]N/OFQ were not affected by naloxone (1 microM) while prevented by the selective NOP receptor antagonist UFP-101 (10 microM). Moreover, the inhibitory effects of NOP agonists were no longer evident in tissues treated with tertiapin (10 microM), an inhibitor of inward-rectifier potassium channels. In conclusion, the present data demonstrate that N/OFQ inhibited acetylcholine (ACh) release in the human bronchi via NOP receptor activation. This effect may involve stimulation of potassium currents. PMID- 16042990 TI - Correlation between the expression of the histone H4 mRNA variant H4-v.1 and the levels of histone H4-(86-100) and H4-(89-102) (OGP) in various rat tissues and alveolar macrophages. AB - We studied the expression of the osteogenic and antinociceptive C-terminal histone H4-related peptide fragments, H4-(89-102) (OGP) and H4-(86-100), respectively, within various rat tissues and isolated alveolar macrophages (AM) by radioimmunoassay (RIA). OGP was located mainly within the bone marrow, spleen, thymus, and lungs whereas H4-(86-100) was more concentrated within the bone marrow, lymph nodes, spinal cord, pituitaries and thymus. The expression pattern of the two peptides showed similarities with the tissue expression pattern of the histone H4 mRNA variant H4-v.1. In rat AM, OGP and H4-(86-100) levels were significantly stimulated (2.6- and 1.9-fold, respectively) by LPS (1 microg/ml), along with H4-v.1 mRNA (4.1-fold), but not whole histone H4 (1.1-fold) nor total histone H4 mRNA (1.1-fold). The results suggest that H4-v.1 mRNA may play a role in the synthesis of the naturally occurring peptides H4-(86-100) and OGP via the alternative translation product H4-(84-102), but not whole histone H4. PMID- 16042991 TI - Suppression of fat deposition for the life time with gene therapy. AB - Unexpended energy is stored as fat in the body and increased rate of fat accretion culminates in obesity. Obesity increases the risks of many diseases several folds and shortens life span. A progressive deficit in the central feedback effects of leptin, a peptide produced by fat cells and hypothalamus, results in increased weight gain and obesity. This article summarizes our experimental findings to show that a stable increase in leptin availability in the hypothalamus alone with the aid of leptin gene therapy suppresses fat accretion and metabolic hormones for nearly the lifetime of laboratory rodents. Consequently, central leptin gene therapy is a novel modality that offers a viable therapeutic option to reduce fat depots and attendant metabolic sequelae implicated in obesity-related illnesses. PMID- 16042992 TI - The influence of the peptide NAP on Mac-1-deficient mice following closed head injury. AB - A single administration of the neuroprotective peptide NAP was previously shown to protect against death associated with closed head injury (CHI) and enhance recovery of the surviving mice. The protective effect was accompanied by down regulation of the relative mRNA content of the complement receptor 3 (Mac-1, a marker for inflammation) as measured about a month after the injury. In contrast, the mRNA transcripts for activity-dependent neuroprotective protein (ADNP, the NAP containing protein) were shown to increase 29 days post CHI in the injured hemisphere of Mac-1 expressing mice. The present study was set out to investigate: (1) are Mac-1-deficient mice less susceptible to the adverse outcome of traumatic head injury; (2) does NAP treatment affect Mac-1-deficient mice subjected to head injury; and (3) is Mac-1 expression associated with ADNP expression. Results showed that (1) Mac-1-deficient mice were partially protected against death associated with severe head injury as compared to Mac-1 expressing mice. (2) Significant protection against death was observed in NAP-treated mice and an increase in recovery was observed in the NAP-treated Mac-1 mice 4 weeks after injury. (3) ADNP expression did not change in the Mac-1-deficient mice following head injury. Our working hypothesis is that a month following injury, gene expression in the injured brain is altered and competing proteins are expressed such as Mac-1 that is associated with inflammation and ADNP that is associated with neuroprotection. Obviously, this plasticity in gene expression is intimately interwoven with the genetic background of the animal. NAP treatment tilts the balance toward neuroprotection. PMID- 16042993 TI - Peptide and non-peptide G-protein coupled receptors (GPCRs) in skeletal muscle. AB - G-protein coupled receptors (GPCRs) represent a large class of cell surface receptors that mediate a multitude of functions. Over the years, a number of GPCRs and ancillary proteins have been shown to be expressed in skeletal muscle. Unlike the case with other muscle tissues like cardiac and vascular smooth muscle cells, there has been little attempt at systematically analyzing GPCRs in skeletal muscle. Here we have compiled all the GPCRs that are expressed in skeletal muscle. In addition, we review the known function of these receptors in both skeletal muscle tissue and in cultured skeletal muscle cells. PMID- 16042994 TI - The relationship between social anxiety disorder and alcohol use disorders: a critical review. AB - Epidemiological studies have demonstrated a significant co-morbidity between social anxiety disorder (SAD) and alcohol use disorders (AUDs). Despite the fact that many studies have demonstrated strong relationships between SAD and AUD diagnoses, there has been much inconsistency in demonstrating causality or even directionality of the relationship between social anxiety and alcohol-related variables. For example, some studies have showed a positive relationship between social anxiety and alcohol-related variables, while others have shown a negative relationship or no relationship whatsoever. In an attempt to better understand the relationship between social anxiety and alcohol, some researchers have explored potential moderating variables such as gender or alcohol expectancies. The present review reports on what has been found with regard to explaining the high co-morbidity between social anxiety and alcohol problems, in both clinical and non-clinical socially anxious individuals. With a better understanding of this complex relationship, treatment programs will be able to better target specific individuals for treatment and potentially improve the efficacy of the treatments currently available for individuals with co-morbid SAD and AUD. PMID- 16042995 TI - Therapeutic and preventive interventions for postulated vasoactive neuropeptide autoimmune fatigue-related disorders. AB - Major advances have been made in understanding the relatively novel group of vasoactive (vasodilatory) neuropeptides (VNs) in humans. VNs comprise a novel but expanding group of substances having immunoregulation, inflammation modulation, neurotransmitter, neurotrophic, hormonal and metabolic functions. These substances may control gene expression for mRNA for themselves and their receptors. They have complex relationships with gaseous and other neurotransmitters and xenobiotic substances. Theoretical arguments have implicated these substances in autoimmune phenomena resulting in fatigue-related conditions such as chronic fatigue syndrome (CFS), sudden infant death syndrome (SIDS), fibromyalgia (FM) and Gulf War syndrome (GWS) but remain unproven. As well as possibly spontaneous onset, the precipitating causes of VN autoimmune dysfunction are likely to be a combination of genetic predisposition, infection and xenobiotic substances. Therapeutic and preventive possibilities for postulated VN autoimmune conditions will be influenced by the complex patholophysiology underpinning them. Some speculative possibilities are VN substitution/replacement, preservation of biological effect, epigenetic DNA modifications, plasma exchange, anti-cholinesterases, e.g., pyridostigmine, corticosteroids and other drug treatments, thymectomy, intravenous immunoglobulin and anti-idiotype antibodies, and CpG/DNA vaccines. Prevention and treatment of possible VN autoimmune fatigue-related disorders may prove to be important areas for future research and development. PMID- 16042996 TI - The incorporation of iodine in thyroid hormone may stem from its role as a prehistoric signal of ecologic opportunity: an evolutionary perspective and implications for modern diseases. AB - To optimize fitness under conditions of varying Darwinian opportunity, organisms demonstrate tremendous plasticity in their life-history strategies based on their perception of available resources. Higher-energy environments generally promote more aggressive life-history strategies, such as faster growth, larger adult size, greater genetic variation, shorter lifespan, larger brood sizes, and offspring ratio skewed towards the larger-sized gender. While numerous mechanisms regulate life-history plasticity including genetic imprinting, methylation, and growth factors, evidence suggests that thyroid hormone plays a central role. Given the pivotal adaptive role of thyroid hormone, the teleology of its dependence on dietary iodine for production remains unexplained. We hypothesize that iodine may have emerged as a substrate for production of thyroid hormone in prehistoric ecosystems because the former represented a reliable proxy for ecologic potential that enabled the latter to modulate growth, reproduction, metabolic rate, and lifespan. Such a scenario may have existed in early marine ecosystems where ocean-surface vegetation, which concentrates iodine for its antimicrobial and antioxidant properties, formed the basis of the food chain. Teleologic parallels can be drawn to the food-chain accumulation of antimicrobials that also exhibit antioxidant properties and promote adult size, brood size, and offspring quality by modulating central hormonal axes. As each higher species in the food chain tunes its life-history strategy based on iodine intake, the coupling of this functional role of iodine with its value as a resource signal to the next member of the food-chain may promote runaway evolution. Whereas predators in prehistoric ecosystems successfully tuned their life-history strategy using iodine as a major input, the strategy may prove maladaptive in modern humans for whom the pattern of iodine intake is decoupled from resource availability. Iodine acquired through sodium iodide supplementation may independently contribute to some biologic dysfunctions currently attributed to sodium. PMID- 16042997 TI - Ketamine use for reduction of opioid tolerance in a 5-year-old girl with end stage abdominal neuroblastoma. PMID- 16042998 TI - Megestrol acetate-probably less effective than has been reported! PMID- 16043000 TI - Lidocaine intoxication at very small doses in terminally ill cancer patients. PMID- 16043001 TI - Mini-dose titration of the transdermal fentanyl patch--a novel approach by adjusting the area of absorption. PMID- 16043002 TI - Family evaluation of hospice care: results from voluntary submission of data via website. AB - The Family Evaluation of Hospice Care (FEHC) survey is a 61-item questionnaire that surveys family members about care provided to the decedent by the hospice. Hospices submit their data to the National Hospice and Palliative Care Organization (NHPCO), where results are tabulated. For the first two quarters of 2004, a total of 29,292 surveys were tabulated. On average, respondents rated their overall satisfaction with care as 47.1 on a 50-point composite scale of five measures of satisfaction. Opportunities for improvement were identified for attending to family needs for support (18.2% of those surveyed reported at least one unmet need), attending to family needs for communication (10-29%), and coordination of care (22.1%). Surrogate reporting of unmet needs for pain, dyspnea, or emotional support ranged from 5.3% to 9.8%. The FEHC is a useful tool for measuring hospice performance and identifies a number of opportunities for improvement. PMID- 16043003 TI - Prospective assessment of patient-rated symptoms following whole brain radiotherapy for brain metastases. AB - To prospectively assess patient-rated symptoms in patients with brain metastases treated with whole brain radiotherapy, these patients were asked to rate their symptoms on the Edmonton Symptom Assessment Scale (ESAS) before, and 1, 2, 4, 8, and 12 weeks following the radiation treatment. ESAS evaluates pain, fatigue, nausea, depression, anxiety, drowsiness, appetite, sense of well-being, and shortness of breath on a scale of 0-10 (0=absence of symptom and 10=worst possible symptom). Patients with a language barrier or significant cognitive impairment were excluded. The mean difference of ESAS symptoms at each follow up were compared with baseline and P < 0.01 was considered statistically significant. One hundred seventy patients (102 female and 68 male) were included between January 1999 and January 2002. Their median age was 66 years (range 33 84) and the median Karnofsky performance score (KPS) at baseline was 60 (range 20 90). The most common primary cancer sites were lung (99; 58%), breast (32; 19%), gastrointestinal (16; 9%), unknown (14; 8%) and others (9; 6%). One-third had significant weight loss (> or =10% over the last 6 months). All patients were prescribed dexamethasone at varying doses during radiotherapy. The dose fractionations were 20 Gy in 5 fractions, 138 (81%); 30 Gy in 10 fractions, 7 (4%); and others, 25 (15%). The baseline mean +/- SD for ESAS scores were: pain 2.4 +/- 2.8, fatigue 5.3 +/- 2.8, nausea 1.3 +/- 2.2, depression 2.8 +/- 2.7, anxiety 3.6 +/- 3.0, drowsiness 3.5 +/- 2.9, appetite 3.0 +/- 3.2, sense of well being 3.8 +/- 2.7, and shortness of breath 2.3 +/- 2.5. For the entire cohort, after the delivery of palliative radiotherapy for brain metastases, there were statistically significant deteriorations in the mean differences from the baseline for the following ESAS domains: fatigue 1.0 to 1.8; drowsiness 1.2 to 1.8; and appetite 2.2 to 2.4. The data demonstrate that certain parameters of quality of life worsen after whole brain radiotherapy. PMID- 16043004 TI - Quality of life in colorectal cancer patients with colectomy and the validation of the Functional Assessment of Cancer Therapy-Colorectal (FACT-C), Version 4. AB - The purpose of this study was to determine whether the Functional Assessment of Cancer Therapy-Colorectal (the FACT-C), a disease-specific tool for the assessment of colorectal cancer patients' QOL, is a valid assessment tool for measuring QOL changes. Ninety-eight colorectal cancer patients performed the assessment at baseline and 52 of these patients completed the instrument at one month and six months after colectomy. In addition to the FACT-C, the shortened forms of the Profile of Mood States, the Eastern Cooperative Oncology Group Performance Status Rating, Neuroticism Scale in the Eysenck Personality Questionnaire, and Functional Living Index-Cancer were completed. We found convergent and divergent validity and good reliability of the FACT-C. Patients' overall QOL was lower at one month after colectomy and recovered to the pre surgery level at six months after colectomy. PMID- 16043005 TI - How people die in hospital general wards: a descriptive study. AB - To describe how patients die in hospital, 370 patients (age >18 years; in hospital for>24 hours) who died on the general wards of 40 Italian hospitals were assessed. Differences between patients whose death was expected and patients whose death was unexpected were evaluated. Data on treatments and care in proximity of death were collected after interviewing the nurse responsible for the patient within 72 hours of the patient's death, and from clinical and nursing records. For 58% of patients, death was highly expected. Symptom control was inadequate for the most severely ill patients: 75% experienced at least one "severe" symptom (42% pain and 45% dyspnea). Nurses tended to judge patients' global care as "good" or "very good" (76%), in spite of the persistence of symptoms and the scant use of analgesics. Despite some encouraging signs of sensitivity to end-of-life problems, acute inpatient institutions in Italy still deal inadequately with the needs of dying persons. PMID- 16043006 TI - Which mini-mental state exam items can be used to screen for delirium and cognitive impairment? AB - Cognitive impairment is common in palliative care patients, but it is frequently undetected. The clinical consequence is that psychiatric states such as delirium, which often present with cognitive impairment, are inadequately treated. A short and simple questionnaire for screening of cognitive impairment is required for these patients, in order to proceed with more advanced testing if necessary. In this study, we explored the results from two samples of patients (n=290 and n=217) who had completed the Mini-Mental State Examination (MMSE). Cases of cognitive impairment are considered indicated by an MMSE score of less than 24 of the total 30. We found that caseness could be fairly accurately screened by using four of the original 20 MMSE items, and that a six-item questionnaire further greatly improved the discrimination. PMID- 16043007 TI - Validation and clinical application of the german version of the palliative care outcome scale. AB - The Palliative Care Outcome Scale (POS) is a multidimensional instrument covering physical, psychosocial, spiritual, organizational, and practical concerns. This study validated the German version of the POS and used the tool in different palliative care settings in Germany and Austria. Patients and staff were asked to complete the POS three times and evaluate the questionnaire and the translation afterwards. One hundred eighteen patients (44 male, 74 female, mean age 63 years, all suffering from advanced cancer) completed the POS one time, 55 patients two times, and 36 patients three times. Spearman's rho was highly significant for pain, other symptoms, anxiety, and life worthwhile in the first two assessments. The third assessment showed significant correlations for pain, other symptoms, anxiety, and family anxiety. Seventy-seven of 87 patients answered questions regarding the scale and the translation. All questions other than "Over the past 3 days, have you felt good about yourself?" were understandable for patients. Almost half of the staff was undecided whether the tool was reflecting the patients' condition. In contrast, the majority of patients liked it. Thus, the German version of the POS is well accepted by patients and staff and appears to be valid, although there are some areas where the scale would benefit from expansion to more closely capture staff and patient concerns. PMID- 16043008 TI - Depressive symptoms, pain experiences, and pain management strategies among residents of taiwanese public elder care homes. AB - The purpose of this study was to explore depressive symptoms, pain experiences, and pain self-care management strategies among residents of public elder care homes in Taiwan. Random sampling was used to recruit participants (n=200). In this sample, the prevalence of depressive symptoms was 49.0%. Pain prevalence was significantly higher in the depressed group (59.2%) than in the non-depressed group (43.1%). Depressed participants tended to report more severe pain intensity, worst pain, average pain, and more interference with walking than the non-depressed group. Most participants (60.0%) took prescribed medications for dealing with pain. Self was the main information source for pain management strategies. Participants reported severe bouts of pain but used limited self-care pain management strategies. Due to the limited number of health care providers in elder care homes, the authors recommend increasing knowledge about depression, pain, and pain management strategies of both institutional health care staff and residents. PMID- 16043009 TI - Home death--the caregivers' experiences. AB - The objectives of this study were to evaluate caregivers' experiences concerning the care of a terminally ill loved one at home, and to compare the death experiences of caregivers with and without access to homecare programs. The primary caregivers of all the patients who died of cancer 6-18 months before the study period (1999-2001) in the Negev area were contacted. This group included 240 caregivers of patients who died in the home palliative care program and 404 caregivers of patients who died with no access to a home palliative care program. A total of 159 caregivers were interviewed, 76 from the home palliative program and 83 who had no access to a palliative care program. Death at home occurred for 80.3% of patients with access to homecare and 20.5% of those without access. Despite the fact that caring for a loved one at home was a greater financial and emotional burden, there was a greater overall satisfaction with the caring experience of those whose loved ones died at home and had access to the homecare program. Given appropriate professional support systems, home-based care at the end of life is preferable to most caregivers. PMID- 16043010 TI - Video-thoracoscopic surgical pleurodesis in the management of malignant pleural effusion: the importance of an early intervention. AB - Thoracentesis plays an important role in cancer patients with symptomatic effusions, although its effect is short-lived and symptoms recur in almost all patients. Early video-thoracoscopic surgical pleurodesis may provide added benefit to a group of patients with advanced cancer presenting with symptomatic malignant pleural effusion. Seventy-six patients with advanced cancer and pleural effusion due to pulmonary-pleural metastases were recruited. In 51 cases (67.1%), at least one thoracentesis was performed before admission for surgery. Preoperative staging consisted of chest radiograph, CT scan, and blood gas analysis. The mean Karnofsky performance status was about 50. Pleurodesis with talc poudrage was completely successful in all patients, with a morbidity rate of 2.6%. There was no post-operative mortality. Three patients (3.9%) underwent further thoracenteses for recurrence of pleural effusion within two months after the procedure. Early use of talc insufflated by video-thoracoscopic surgery is an effective and relatively safe method for treating pleural effusion, and preventing recurrence, in advanced cancer patients. PMID- 16043011 TI - Compatibility and stability of dexamethasone sodium phosphate and ketamine hydrochloride subcutaneous infusions in polypropylene syringes. AB - The stability of ketamine hydrochloride injection and dexamethasone sodium phosphate injection, when mixed and stored in polypropylene syringes, was studied. Formulations containing ketamine hydrochloride (50 mg or 600 mg) and dexamethasone sodium phosphate (1 mg) in 0.9% sodium chloride injection (to 14 ml) were prepared and stored at 4 degrees C, 23 degrees C, and 37 degrees C, under normal fluorescent light conditions, for 192 hours. The concentrations of the drugs were determined at 0, 2, 4, 8, 24, 48, 96, and 192 hours using a validated high-pressure liquid chromatography method. The pH, color, and visible particles of each solution were also assessed at each time point. All formulations tested maintained more than 98% of the initial concentrations of both drugs, and no degradation products were detected. The solutions remained clear and colorless and the pH varied within 0.05 units throughout 192 hours. The results indicate that, at the concentrations studied, combinations of ketamine hydrochloride and dexamethasone sodium phosphate in 0.9% sodium chloride injection were physically and chemically stable for at least 192 hours (8 days) when stored in polypropylene syringes. PMID- 16043012 TI - Percutaneous vertebroplasty--a technique to treat refractory spinal pain in the setting of advanced metastatic cancer: a case series. AB - We present three cases of severe movement-related spinal pain in patients with advanced metastatic carcinomas successfully treated with percutaneous vertebroplasty (PV). These patients had multi-symptom burden and progressive metastasis. Their movement-related pain was incapacitating and refractory to a variety of more conservative interventions. PV is a minimally invasive technique to stabilize vertebral compression fractures, thereby decreasing spinal pain in this setting. Its use in the setting of advanced cancer with severe movement related pain has not been previously clarified in the palliative care literature. In summary, PV is a technique with a favorable risk: benefit ratio even in the setting of advanced metastatic cancer. The keys to successful utilization of PV in this patient population are careful patient assessment and selection as outlined in the report, in addition to an experienced care team approach. PMID- 16043013 TI - Opioid rotation from morphine to fentanyl in delirious cancer patients: an open label trial. AB - Although recent studies suggest that opioid rotation could be an effective treatment strategy for morphine-induced delirium, there have been no prospective studies to investigate the treatment effects of opioid rotation using fentanyl. The primary aim of this study was to clarify the efficacy of opioid rotation from morphine to fentanyl in symptom palliation of morphine-induced delirium. Twenty one consecutive cancer patients with morphine-induced delirium underwent opioid rotation to fentanyl. Physicians recorded the symptom severity of delirium (the Memorial Delirium Assessment Scale, MDAS), pain, and other symptoms (categorical verbal scale from 0: none to 3: severe) and the Schedule for Team Assessment Scale (STAS) (from 0: none to 4: extreme); and performance status at the time of study enrollment and three and seven days after. Of 21 patients recruited, one patient did not complete the study. In the remaining 20 patients, morphine was substituted with transdermal fentanyl in 9 patients and parenteral fentanyl in 11 patients. Total opioid dose increased from 64 mg oral morphine equivalent/day (Day 0) to 98 mg/day (Day 7), and the median increase in total opioid dose was 42%. Treatment success, defined as an MDAS score below 10 and pain score of 2 or less, was obtained in 13 patients on Day 3 and 18 patients on Day 7. The mean MDAS score significantly decreased from 14 (Day 0) to 6.4 and 3.6 (Days 3 and 7, respectively, P < 0.001). Pain scores significantly decreased from 2.2 (Day 0) to 1.3 and 1.1 on the categorical verbal scale (Days 3 and 7, respectively, P < 0.001); from 2.6 (Day 0) to 1.6 and 1.3 on the STAS (Days 3 and 7, respectively, P < 0.001). Symptom scores of dry mouth, nausea, and vomiting significantly decreased, and performance status significantly improved. Opioid rotation from morphine to fentanyl may be effective in alleviating delirium and pain in cancer patients with morphine-induced delirium. PMID- 16043014 TI - Implementation of a massage therapy program in the home hospice setting. PMID- 16043015 TI - Antioxidant strategies in the treatment of stroke. AB - Excessive production of free radicals is known to lead to cell injury in a variety of diseases, such as cerebral ischemia. In this review, we describe some of the numerous studies that have examined this oxidative stress and the efficiency of antioxidant strategies in focal cerebral ischemia. Besides using genetically modified mice, these strategies can be divided into three groups: (1) inhibition of free radical production, (2) scavenging of free radicals, and (3) increase of free radical degradation by using agents mimicking the enzymatic activity of endogenous antioxidants. Finally, the clinical trials that have tested or are currently testing the efficiency of antioxidants in patients suffering from stroke are reviewed. The results presented here lead us to consider that antioxidants are very promising drugs for the treatment of ischemic stroke. PMID- 16043016 TI - Statin attenuates high glucose-induced and diabetes-induced oxidative stress in vitro and in vivo evaluated by electron spin resonance measurement. AB - An increased oxidative stress may contribute to the accelerated atherosclerosis in diabetic patients. Here we show that 3-hydroxy-3-methylglutaryl CoA reductase inhibitor (statin) attenuates a high glucose-induced and a diabetes-induced oxidative stress through inhibition of vascular NAD(P)H oxidase. Exposure of cultured aortic endothelial cells and smooth muscle cells to a high glucose level (450 mg/dl) for 3 days significantly increased oxidative stress compared with a normal glucose level (100 mg/dl), as evaluated by the staining with 2',7' dichlorofluorescein diacetate and electron spin resonance (ESR) measurement. This increase was completely blocked by the treatment with pitavastatin (5 x 10(-7)M) as well as a NAD(P)H oxidase inhibitor (diphenylene iodonium) or a PKC inhibitor (calphostin C) in parallel with the change of small GTPase Rac-1 activity, a cytosolic regulatory component of NAD(P)H oxidase. Next, using streptozotocin induced diabetic rats, the effect of pitavastatin on oxidative stress was evaluated by in vivo ESR measurements, which is a sensitive, noninvasive method. Administration of pitavastatin (5 mg/kg/day) for 4 days attenuated the increased oxidative stress in diabetic rats to control levels. In conclusion, pitavastatin attenuated a high glucose-induced and a diabetes-induced oxidative stress in vitro and in vivo. Thus, our data may provide a new insight into antioxidative therapy in diabetes. PMID- 16043017 TI - Redox proteomics analysis of oxidatively modified proteins in G93A-SOD1 transgenic mice--a model of familial amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron degenerative disease characterized by the loss of neuronal function in the motor cortex, brain stem, and spinal cord. Familial ALS cases, accounting for 10-15% of all ALS disease, are caused by a gain-of-function mutation in Cu,Zn-superoxide dismutase (SOD1). Two hypotheses have been proposed to explain the toxic gain of function of mutant SOD (mSOD). One is that mSOD can directly promote reactive oxygen species and reactive nitrogen species generation, whereas the other hypothesis suggests that mSODs are prone to aggregation due to instability or association with other proteins. However, the hypotheses of oxidative stress and protein aggregation are not mutually exclusive. G93A-SOD1 transgenic mice show significantly increased protein carbonyl levels in their spinal cord from 2 to 4 months and eventually develop ALS-like motor neuron disease and die within 5-6 months. Here, we used a parallel proteomics approach to investigate the effect of the G93A-SOD1 mutation on protein oxidation in the spinal cord of G93A-SOD1 transgenic mice. Four proteins in the spinal cord of G93A-SOD1 transgenic mice have higher specific carbonyl levels compared to those of non-transgenic mice. These proteins are SOD1, translationally controlled tumor protein (TCTP), ubiquitin carboxyl terminal hydrolase-L1 (UCH-L1), and, possibly, alphaB-crystallin. Because oxidative modification can lead to structural alteration and activity decline, our current study suggests that oxidative modification of UCH-L1, TCTP, SOD1, and possibly alphaB-crystallin may play an important role in the neurodegeneration of ALS. PMID- 16043018 TI - Inhaled glutathione decreases PGE2 and increases lymphocytes in cystic fibrosis lungs. AB - Reduced glutathione (GSH), a major antioxidant and modulator of cell proliferation, is decreased in the bronchoalveolar lavage fluid (BALF) of cystic fibrosis (CF) patients. We previously have shown that GSH inhalation in CF patients significantly increased GSH levels in BALF and improved lung function (M. Griese et al., 2004, Am. J. Respir. Crit. Care Med.169, 822-828). GSH depletion in vitro enhances susceptibility to oxidative stress, increases inflammatory cytokine release, and impairs T cell responses. We therefore hypothesized that an increase in GSH in BALF reduces oxidative stress, decreases inflammation, and modulates T cell responses in lungs of CF patients. BALF from 17 CF patients (median FEV1 67% (43-105%) of predicted) was assessed before and after GSH inhalation for total protein, markers of oxidative stress (8 isoprostane, myeloperoxidase, and ascorbic and uric acid), pattern of protein oxidation, prostaglandin E2 (PGE2), and proinflammatory cytokines. BALF cells were differentiated using cytospin slides, and lymphocytes were further analyzed by flow cytometry. Inhalation of GSH decreased BALF levels of PGE2 and increased CD4+ and CD8+ lymphocytes in BALF significantly but had no effect on markers of oxidative stress. BALF lymphocytes correlated positively with lung function, whereas levels of PGE2 showed an inverse correlation. The patients with the greatest improvement in lung function after GSH treatment also had the largest decline in PGE2 levels. We conclude that GSH inhalation in CF patients increases lymphocytes and suppresses PGE2 in the bronchoalveolar space. Thus, GSH primarily affected the pulmonary immune response rather than the oxidative status in CF patients. The effect of GSH inhalation on PGE2 levels and lymphocytes in CF warrants further investigation. PMID- 16043019 TI - Cytotoxicity and genotoxicity induced by the photochemical alkoxyl radical source N-tert-butoxypyridine-2-thione in L5178Y mouse lymphoma cells under UVA irradiation. AB - The cell-damaging effects of N-tert-butoxypyridine-2-thione (tBuOPT), which generates alkoxyl and thiyl radicals on photolysis, have been investigated in L5178Y mouse lymphoma cells. The UVA irradiation of 2.5 microM tBuOPT inhibits strongly cell growth and cell viability, causes pronounced membrane damage, and induces micronuclei. Without irradiation, tBuOPT does not cause any cell damage at 2.5 microM concentration. The phototoxicity of tBuOPT is effectively inhibited by the radical scavenger glutathione, while the photogenotoxicity (micronuclei induction) is not affected by this strong hydrogen-atom donor. Thus, for the cytotoxicity and genotoxicity different reactive species seems to be responsible. The cytotoxicity is presumably caused by oxyl radicals, which are derived from tert-butoxyl radicals generated by photocleavage of tBuOPT, while in the genotoxicity the less reactive pyridyl-2-thiyl radicals appear to play a role. These results demonstrate that N-alkoxypyridinethiones are useful photochemical sources of oxyl and thiyl radicals to elucidate biological effects caused by these free radicals. PMID- 16043020 TI - Nitration of gamma-tocopherol prevents its oxidative metabolism by HepG2 cells. AB - Gamma-tocopherol (gammaT) is one of the major forms of vitamin E consumed in the diet. Previous reports have suggested increased levels of nitrated gamma tocopherol (5-NO2-gammaT) in smokers and individuals with conditions associated with elevated nitrative stress. The monitoring of 5-NO2-gammaT and its possible metabolite(s) may be a useful marker of reactive nitrogen species generation in vivo. The major pathway for the metabolism of gammaT is the cytochrome P450 dependent oxidation to its water-soluble metabolite gamma-CEHC, which is excreted in urine. In order to determine if 5-NO2-gammaT could be metabolised via the same route and detected in urine we developed a sensitive gas chromatography-mass spectrometry assay for 5-NO2-gamma-CEHC. 5-NO2-gamma-CEHC was synthesised and its structure confirmed by proton nuclear magnetic resonance and mass spectrometry. While gamma-CEHC was abundant in urine from healthy volunteers, as well as patients with coronary heart disease and type 2 diabetes, 5-NO2-gamma-CEHC was undetectable (limit of detection of 5 nM). To understand this observation we examined the uptake and metabolism of gammaT and 5-NO2-gammaT by HepG2 cells. gammaT was readily incorporated into cells and metabolised to gamma-CEHC over a period of 48 hours. In contrast, 5-NO2-gammaT was poorly incorporated into HepG2 cells and not metabolised to 5-NO2-gamma-CEHC over the same time period. We conclude that nitration of gammaT prevents its incorporation into liver cells and therefore its metabolism to the water-soluble metabolite. Whether 5-NO2-gammaT could be metabolised via other pathways in vivo requires further investigation. PMID- 16043021 TI - Cinnamophilin reduces oxidative damage and protects against transient focal cerebral ischemia in mice. AB - Acute neuroprotective effects of cinnamophilin (CINN; (8R, 8'S)-4, 4'-dihydroxy 3, 3'-dimethoxy-7-oxo-8, 8'-neolignan), a novel antioxidant and free radical scavenger, were studied in a mouse model of transient middle cerebral artery (MCA) occlusion. CINN was administered intraperitoneally either 15 min before (pretreatment) or 2 h after the onset of MCA occlusion (postischemic treatment). Relative to vehicle-treated controls, animals pretreated with CINN, at 20-80 mg/kg, had significant reductions in brain infarction by 33-46% and improvements in neurobehavioral outcome. Postischemic administration with CINN (80 mg/kg) also significantly reduced brain infarction by 43% and ameliorated neurobehavioral deficits. Additionally, CINN administration significantly attenuated in situ accumulation of superoxide anions (O2-) in the boundary zones of infarct at 4 h after reperfusion. Consequently, CINN-treated animals exhibited significantly decreased levels of oxidative damage, as assessed by immunopositive reactions for 8-hydroxy-2'-deoxyguanosine (8-OHdG) and 4-hydroxynonenal (4-HNE), and the resultant inflammatory reactions at 24 h post-insult. It is concluded that CINN effectively reduced brain infarction and improved neurobehavioral outcome following a short-term recovery period after severe transient focal cerebral ischemia in mice. The finding of a decreased extent of reactive oxygen species and oxidative damage observed with CINN treatment highlights that its antioxidant and radical scavenging ability is contributory. PMID- 16043022 TI - Intra- and intermolecular oxidation of oxymyoglobin and oxyhemoglobin induced by hydroxyl and carbonate radicals. AB - The mechanism of the reactions of myoglobin and hemoglobin with *OH and CO3*- in the presence of oxygen was studied using pulse and gamma-radiolysis. Unlike *NO2, which adds to the porphyrin iron, *OH and CO3*- form globin radicals. These secondary radicals oxidize the Fe(II) center through both intra- and intermolecular processes. The intermolecular pathway was further demonstrated when BSA radicals derived from *OH or CO3*- oxidized oxyhemoglobin and oxymyoglobin to their respective ferric states. The oxidation yields obtained by pulse radiolysis were lower compared to gamma-radiolysis, where the contribution of radical-radical reactions is negligible. Full oxidation yields by *OH-derived globin radicals could be achieved only at relatively high concentrations of the heme protein mainly via an intermolecular pathway. It is suggested that CO3*- reaction with the protein yields Tyr and/or Trp-derived phenoxyl radicals, which solely oxidize the porphyrin iron under gamma-radiolysis conditions. The *OH particularly adds to aromatic residues, which can undergo elimination of H2O forming the phenoxyl radical, and/or react rapidly with O2 yielding peroxyl radicals. The peroxyl radical can oxidize a neighboring porphyrin iron and/or give rise to superoxide, which neither oxidize nor reduce the porphyrin iron. The potential physiological implications of this chemistry are that hemoglobin and myoglobin, being present at relatively high concentrations, can detoxify highly oxidizing radicals yielding the respective ferric states, which are not toxic. PMID- 16043023 TI - Reduction of antigen-induced respiratory abnormalities and airway inflammation in sensitized guinea pigs by a superoxide dismutase mimetic. AB - Reactive oxygen species have been implicated in the pathogenesis of asthma and, in atopic asthmatics, endogenous superoxide dismutase (SOD) enzyme levels are known to decrease. This suggests that replacing a failed endogenous SOD enzyme system with a mimetic of the endogenous enzyme would be beneficial and protective. In this study we demonstrate that removal of superoxide by the SOD mimetic (SODm) M40403 reduces the respiratory and histopathological lung abnormalities due to ovalbumin (OA) aerosol in a model of allergic asthma-like reaction in sensitized guinea pigs. Both respiratory abnormalities and bronchoconstriction in response to OA challenge are nearly absent in naive animals, while they sharply became severe in sensitized animals. In addition, OA aerosol induced a reduction of MnSOD activity which was paralleled with bronchiolar lumen reduction, pulmonary air space hyperinflation, mast cell degranulation, eosinophil infiltration, bronchial epithelial cell apoptosis, increase in myeloperoxidase activity, malonyldialdehyde production and 8-hydroxy 2'-deoxyguanosine formation in the lung tissue, as well as elevation of PGD2 in the bronchoalveolar lavage fluid. Treatment with M40403 restored the levels of MnSOD activity and significantly reduced all the above parameters. In summary, our findings support the potential therapeutic use of SOD mimetics in asthma and anaphylactic reactions and account for a critical role for superoxide in acute allergic asthma-like reaction in actively sensitized guinea pig. PMID- 16043024 TI - Nitrated lipids decompose to nitric oxide and lipid radicals and cause vasorelaxation. AB - Nitric oxide-derived oxidants such as nitrogen dioxide and peroxynitrite have been receiving increasing attention as mediators of nitric oxide toxicity. Indeed, nitrated and nitrosated compounds have been detected in biological fluids and tissues of healthy subjects and in higher yields in patients under inflammatory or infectious conditions as a consequence of nitric oxide overproduction. Among them, nitrated lipids have been detected in vivo. Here, we confirmed and extended previous studies by demonstrating that nitrolinoleate, chlolesteryl nitrolinoleate, and nitrohydroxylinoleate induce vasorelaxation in a concentration-dependent manner while releasing nitric oxide that was characterized by chemiluminescence-and EPR-based methodologies. As we first show here, diffusible nitric oxide production is likely to occur by isomerization of the nitrated lipids to the corresponding nitrite derivatives that decay through homolysis and/or metal ion/ascorbate-assisted reduction. The homolytic mechanism was supported by EPR spin-trapping studies with 3,5-dibromo-4 nitrosobenzenesulfonic acid that trapped a lipid-derived radical during nitrolinoleate decomposition. In addition to provide a mechanism to explain nitric oxide production from nitrated lipids, the results support their role as endogenous sources of nitric oxide that may play a role in endothelium independent vasorelaxation. PMID- 16043025 TI - Evidence of cardiovascular protection by moderate alcohol: role of nitric oxide. AB - Epidemiological evidence indicates that moderate alcohol consumption reduces the incidence of heart disease. Endothelial nitric oxide synthase (eNOS) is a key regulator of vascular homeostasis and myocardial functions through the controlled production of nitric oxide (*NO). These studies were conducted to determine if the apparent alcohol-associated cardioprotection is mediated, in part, through modulation of the eNOS protein and activity in the cardiovascular system. Rats were fed alcohol and eNOS protein and *NO production were evaluated at the end of 8 weeks. Myocardial and vascular function was assessed ex vivo in a subset of animals. Moderate alcohol improved postischemic myocardial systolic and diastolic function and attenuated the postischemic reduction in coronary vascular resistance. Moderate alcohol also enhanced maximum vascular relaxation by 26 +/- 0.2% and increased plasma *NO production concomitant with a greater than 2.5-fold increase in eNOS protein. Higher levels of alcohol impaired maximum vascular relaxation by 22 +/- 0.1%. These results suggest that moderate alcohol improves postischemic myocardial functions and increases *NO production by vascular endothelium. An increase in *NO may explain, at least in part, the cardioprotective benefits of moderate alcohol consumption. PMID- 16043026 TI - Effect of antioxidant-enriched diets on glutathione redox status in tissue homogenates and mitochondria of the senescence-accelerated mouse. AB - The main purpose of this study was to investigate whether consumption of diets enriched in antioxidants attenuates the level of oxidative stress in the senescence-accelerated mouse (SAM). In separate and independent studies, two different dietary mixtures, one enriched with vitamin E, vitamin C, L-carnitine, and lipoic acid (Diet I) and another diet including vitamins E and C and 13 additional ingredients containing micronutrients with bioflavonoids, polyphenols, and carotenoids (Diet II), were fed for 8 and 10 months, respectively. The amounts of glutathione (GSH) and glutathione disulfides (GSSG) and GSH:GSSG ratios were determined in plasma, tissue homogenates, and mitochondria isolated from five different tissues of SAM (P8) mice. Both diets had a reductive effect in plasma; however Diet I had relatively little effect on the glutathione redox status in tissue homogenates or mitochondria. Remarkably, Diet II caused a large increase in the amount of glutathione and a marked reductive shift in glutathione redox state in mitochondria. Overall, the effects of Diet II were tissue and gender specific. Results indicated that the glutathione redox state in mitochondria and tissues can be altered by supplemental intake of a relatively complex mixture of dietary antioxidants that contains substances known to induce phase 2 enzymes, glutathione, and antioxidant defenses. Whether corresponding attenuations occur in age-associated deleterious changes in physiological functions or life span remains unknown. PMID- 16043027 TI - Catalytically inactive heme oxygenase-2 mutant is cytoprotective. AB - Heme oxygenase (HO) catalyzes the rate-limiting step in heme degradation, producing iron, carbon monoxide, and bilirubin/biliverdin. HO consists of two isozymes: HO-1, which is an oxidative stress-response protein, and HO-2, which is constitutively expressed. HO-2 accounts for most HO activity within the nervous system. Its posttranslational modifications and/or interactions with other proteins make HO-2 a unique regulator of cellular homeostasis. Our previous results revealed that brain infarct volume was enlarged in HO-2 knockout mice. A similar neuroprotective role of HO-2 was shown using primary cortical neurons. To better understand the neuroprotective mechanism of HO-2, we used a catalytically inactive mutant, HO-2H45A, and investigated its cellular effects in response to hemin and hydrogen peroxide-induced cytotoxicity. We observed that HO-2WT overexpression in the HEK293 cell lines became less sensitive to hemin, whereas the inactive mutant HO-2H45A was more sensitive to hemin as compared to control. Interestingly, HO-2WT- and HO-2H45A-overexpressing cells were both protected against H2O2-induced oxidative stress and had less oxidatively modified proteins as compared to control cells. These data indicate that when HO-2 cannot metabolize the prooxidant heme, more cytotoxicity is found, whereas, interestingly, the catalytically inactive HO-2H45A was also able to protect cells against oxidative stress injury. These results suggest the multiplicity of action of the HO-2 protein itself. PMID- 16043028 TI - Resveratrol as an anticancer nutrient: molecular basis, open questions and promises. AB - The polyphenol resveratrol is an anticancer nutrient that was shown to inhibit cancer initiation and promotion [Jang M, Cai L, Udeani GO, Slowing KV, Thomas CF, Beecher CW, et al. Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science 1997;275:218-20]. The absorption, transport and metabolism of resveratrol will be reviewed as well as its actions in multiple pathways involved in the regulation of the cell cycle and the induction of apoptosis. Resveratrol acts as a selective estrogen receptor modulator (SERM) and regulates proteins involved in DNA synthesis and cell cycle, such as p(53) and Rb/E2F, cyclins, cyclin-dependent kinases (CDKs) and their inhibitors. Resveratrol affects the activity of transcriptional factors involved in proliferation and stress responses, such as NF-kB, AP1 and Egr1. Part of these events is mediated by mitogen-activated protein kinases (MAPKs) and tyrosine kinases (e.g., Src) and leads to the modulation of survival and apoptotic factors [e.g., Bcl2 family members, inhibitors of apoptosis (IAPs), ceramide] as well as enzymes involved in carcinogenesis [cyclooxygenases (COXs), nitric oxide synthase (NOS), phase I and II enzymes]. Moreover, resveratrol affects the expression and the activity of cotranscriptional factors such as p(300) and sirtuin 1. Thus, resveratrol potential as an anticancer chemopreventive and chemotherapeutic agent and its implication in the prosurvival versus prodeath pathway induction will be discussed. PMID- 16043029 TI - Uracil misincorporation into DNA of leukocytes of young women with positive folate balance depends on plasma vitamin B12 concentrations and methylenetetrahydrofolate reductase polymorphisms. A pilot study. AB - Changes in the folate and vitamin B12 status in the body influence the extent of uracil misincorporation (UrMis) into DNA, which is one of the biomarkers of genomic stability and, thus, portends a risk of cancer. In our study, the level of UrMis into DNA was evaluated by the comet assay (using the specific DNA repair enzyme, uracil DNA glycosylase) in leukocytes from blood donated by healthy young women with positive folate balance achieved by 4 weeks of folic acid supplementation (400 microg/day). The nutritional status was evaluated on the basis of nine food diaries recorded by the subjects during two winter months. The data were computerized, and the intake of nutrients and micronutrients was estimated using the DIETA 2 program (Food and Nutrition Institute, Warsaw, Poland) linked to recently updated Polish food tables. The plasma folate and vitamin B12 concentration, as well as methylenetetrahydrofolate reductase (MTHFR) polymorphisms, were evaluated to determine their influence on the level of UrMis into DNA. The mean value of B12 intake for all subjects reached 100% of the Polish recommended dietary allowances (RDA), whereas the mean value of folate intake, before folate supplementation, was 50%, suggesting moderate deficiency. Folic acid supplementation brought the folate intake way above the RDA, and plasma folate concentration in each individual was above the deficient range (mean value 14.67 ng/ml). The UrMis did not correlate with the plasma folate concentration, but the level of UrMis was significantly lower in subjects with plasma vitamin B12 concentration above 400 pg/ml (P=.02) only after folic acid supplementation. The concentration of folate in plasma correlated (P1800 counts/s, centered at 490 nm) is observed just slightly to the red of the J aggregate absorption maximum. In the case of cetyltrimethyl-ammonium bromide, increasing surfactant concentrations have only shown to favor solubilization of porphyrin monomers. Evidently, the nature of polar headgroups of surfactants influences the tendency of THPP to aggregate. PMID- 16043041 TI - Single crystal EPR study of VO(II) in magnesium potassium phosphate hexahydrate: a case of two substitutional vanadyl ions. AB - Single crystal EPR study of VO(II)-doped magnesium potassium phosphate hexahydrate has been carried out at room temperature. Single crystal rotations in the three orthogonal planes indicate that the paramagnetic impurity has entered the lattice in place of magnesium ions by replacing the equatorial and axial coordinated water molecules. The spin Hamiltonian parameters obtained for the two substitutional sites are: site 1: g1=1.9800, g2=1.9774, g3=1.9296, A1=7.25 mT, A2=8.09 mT, A3=18.69 mT and site 2: g1=1.9802, g2=1.9765, g3=1.9296, A1=6.82 mT, A2=8.48 mT, A3=18.75 mT, respectively. Superhyperfine, from protons of ligand water molecules, has been observed at certain orientations. It has been concluded that VO bond directions of the two substitutional ions are orthogonal to each other. PMID- 16043042 TI - Synthesis, characterization and photophysical processes of fluorene derivative containing naphthalene nucleus. AB - A novel luminescent compound, 9,9-bis[4'-(beta-naphthyl methacrylate)phenyl]fluorene (F-NMAP) is synthesized by Heck reaction of beta naphthyl methacrylic ester (NMAE) and 9,9-bis(4'-iodophenyl)-fluorene (BIPF). The structure is characterized by MS, 1H NMR, IR, and UV-vis spectroscopy. The photophysical processes of F-NMAP have been carefully investigated by UV-vis absorption and fluorescence spectra. The results show that the compound emits blue and blue-violet light. The luminescence quantum yield is 0.652 in ethanol and the emission spectra exhibit obvious solvent effect. With the increase in polarity of solvents, the fluorescence spectra change obviously and appear blue shift at room temperature. The light emitting can be quenched by both electron donor, N,N-dimethylaniline (DMA), and electron acceptor, dimethylterephthalate (DMTP). On adding gradually, DMA or DMTP into the solution of F-NMAP, the emission intensities of fluorescence are unusually increased. But when the concentration of DMA or DMTP goes beyond a certain scope, the emission intensities of fluorescence are gradually decreased. Simultaneously, the maximum emission peaks of F-NMAP-added DMA are blue-shifted and the maximum emission peaks of F-NMAP-added DMTP are red-shifted, respectively. Moreover, interactions between F-NMAP and fullerene (C60), or carbon nanotubes (CNTs) are also studied by fluorescent quenching where the processes follow the Stern-Volmer equation. PMID- 16043043 TI - DFT studies, vibrational spectra and conformational stability of 4-hydroxy-3 methylacetophenone and 4-hydroxy-3-methoxyacetophenone. AB - The FT-IR and FT-Raman spectra of 4-hydroxy-3-methylacetophenone (HMA) and 4 hydroxy-3-methoxyacetophenone (HMOA) have been recorded. The total energy calculations of HMA and HMOA were tried for various possible conformers. The spectra were interpreted with the aid of normal coordinate analysis based on density functional theory (DFT) using standard B3LYP/6-31G* and B3LYP/6-311+G** methods and basis sets combinations for the most optimized geometries. Normal coordinate calculations were performed with the DFT force field corrected by a recommended set of scaling factors yielding fairly good agreement between observed and calculated frequencies. On the basis of the comparison between calculated and experimental results, assignments of fundamental modes were examined. PMID- 16043044 TI - Vibrational spectra and potential energy distributions for 4,5-dichloro-3 hydroxypyridazine by density functional theory and normal coordinate calculations. AB - The solid phase FT-IR and FT-Raman spectra of 4,5-dichloro-3-hydroxypyridazine have been recorded in the regions 4000-400 cm(-1) and 3500-100 cm(-1), respectively. The spectra were interpreted with the aid of normal coordinate analysis following a full structure optimization and force field calculations based on the density functional theory (DFT) using the standard B3LYP/6-31G* and B3LYP/6-311+G** method and basis set combinations. The DFT force field transformed to natural internal coordinates was corrected by a well-established set of scale factors that were found to be transferable to the title compound. The IR and Raman spectra were predicted theoretically and compared with the experimental spectra. PMID- 16043045 TI - A highly sensitive assay for spectrofluorimetric determination of reduced glutathione using organic nano-probes. AB - In this study, the new nanometer-sized fluorescent particles (1-pyrenemethylamine nanoparticles) have been prepared by reprecipitation method under ultrasonic radiation. These nanoparticles have the potential to overcome problems encountered by organic small molecules by combining the advantages of high photobleaching threshold, high quantum yield, long fluorescence lifetime, good chemical stability, and wide excitation spectral properties. These nanoparticles will be able to be directly used as fluorescent nanoparticles probe without modification. A new fluorimetric method for the determination of reduced glutathione (GSH) has been developed with these nanoparticles. Under optimal conditions, the organic nanoparticles reacted with GSH and o-phthalaldehyde (OPA) to give a highly fluorescent derivative in Na2CO3-HCl buffer (pH=9.0). The fluorescence excitation and emission wavelengths of fluorescent derivative were located at 345 and 400 nm, respectively. The relative fluorescence intensity (RF) was linear in the range of the GSH concentration from 8.0x10(-7) to 1.1x10( 4)moll(-1). Limit of detection of 7.1x10(-8)moll(-1) was achieved for the reduced glutathione. The method was validated and applied to the analysis of three synthetic samples containing reduced glutathione. PMID- 16043046 TI - Spectroscopic modification of the Pippard relation applied for the translational mode in ammonia solid II near the melting point. AB - This study gives our calculation for the frequency shifts 1v partial differentialv partial differentialT(P) and the specific heat C(P) near the melting point in the ammonia solid II. We establish a linear relationship between C(P) and 1v partial differentialv partial differentialT(P) using the Raman frequencies of the translational mode which we calculated in this system. This leads to the validity of the spectroscopic modification of the first Pippard relation in the ammonia solid II near the melting point. From this linear variation of C(P) with the 1v partial differentialv partial differentialT(P) we deduce the slope values of dP(m)dT near the melting point for the fixed pressures of 3.65, 5.02 and 6.57 kbar in the ammonia solid II. They are compared with the experimental dP(m)dT values for this system. PMID- 16043047 TI - Spectroscopic evaluation of manganese(II) complexes derived from semicarbazones and thiosemicarbazones. AB - Manganese(II) complexes having the general composition Mn(L)2X2 [where L=isopropyl methyl ketone semicarbazone (LLA), isopropyl methyl ketone thiosemicarbazone (LLB), 4-aminoacetophenone semicarbazone (LLC) and 4 aminoacetophenone thiosemicarbazone (LLD) and X=Cl-, 1/2SO(4)2-] have been synthesized. All the complexes were characterized by elemental analyses, molar conductance, magnetic moment susceptibility, EI-mass, 1H NMR, IR, EPR and electronic spectral studies. All the complexes show magnetic moments corresponding to five unpaired electrons. The possible geometries of the complexes were assigned on the basis of EPR, electronic and infrared spectral studies. PMID- 16043048 TI - Near IR overtone spectral investigations of cyclohexanol using local mode model- evidence for variation of anharmonicity with concentration due to hydrogen bonding. AB - The near infrared vibrational overtone absorption spectrum of liquid phase cyclohexanol in carbon tetrachloride in different concentrations are examined in the region Deltav=2, 3 and 4. The free and bonded OH local mode mechanical frequency values and anharmonicity values obtained from fitting the overtones are analysed. The observation supports the conclusions drawn from earlier experimental studies on anharmonicity variation of OH-stretching vibrations of alcohols due to intermolecular hydrogen bonding. Our observation is also in agreement with the ab initio calculations on water dimer and trimer. Mechanical anharmonicity of bonded OH-stretching bands tends to increase as a consequence of strong hydrogen bonding at higher concentrations. PMID- 16043049 TI - Theoretical prediction of gas-phase infrared spectra of imidazo[1,2 a]pyrazinediones and imidazo[1,2-a]imidazo[1,2-d]pyrazinediones derived from glycine. AB - Imidazo[1,2-a]pyrazine-3,6-diones and imidazo[1,2-a]imidazo[1,2-d]pyrazine-3,8 diones can be produced by pyrolysis of simple amino acids. While such bicyclic and tricyclic amidines were detected and characterized by IR spectroscopy for some alpha-substituted amino acids, the parent systems composed of glycine fragments are unknown up to now. IR spectra for five amidines derived from glycine were calculated by using different semi-empirical (PM3, AM1, MNDO and MINDO/3), HF, and hybrid DFT (B3LYP, B3P86 and B3PW91) methods in conjunction with 6-31G(d) basis set (for HF and DFT). Vibration frequencies in the experimental IR spectra were predicted based upon the B3LYP data, by correcting the calculated wavenumbers by a scaling factor of 0.959. The behavior of most characteristic bands (nu(CX), nu(NH), etc.) and their shifts with respect to such bands in the spectra of alanine and alpha-aminoisobutyric acid derivatives studied before, are discussed. Performance of the semi-empirical methods was tested, bearing in mind possible future needs for IR spectra predictions for larger molecular systems of similar chemical nature; the use of MINDO/3 and MNDO is recommended. A basis set effect on the B3LYP fundamental vibration frequencies for hexahydroimidazo[1,2-a]pyrazine-3,6-dione was studied by varying Pople basis sets from minimal STO-3G to 6-311++G(d, p). No significant improvements were found beyond the 6-31G(d) basis set, which thus can be recommended to predict IR spectra for the amidines and similar molecules. PMID- 16043050 TI - Infrared spectra of metal-free, N',N-dideuterio, and magnesium porphyrins: density functional calculations. AB - Infrared frequencies and intensities for the metal-free porphyrin (H2Por), N',N dideuterio porphyrin (D2Por), and magnesium porphyrin complex (MgPor) have been calculated at density functional B3LYP level using the 6-31G(d) basis set. Detailed assignments of the NH and NM vibrational bands in the IR spectra have been made on the basis of comparison between the calculated data and the experimental results. The previous empirical controversial assignments for the bands at 1224, 1110 and 771 cm(-1) for metal-free porphyrin are also clearly interpreted. PMID- 16043051 TI - Study for luminescence performance of three methyl xanthine derivatives. AB - In this paper, the low-temperature phosphorescence (LTP), the low-temperature fluorescence (LTF), the paper substrate room-temperature phosphorescence (PS-RTP) and the room fluorescence (RTF) properties of caffeine (CF), theophylline (TP), and theobromine (TB) are investigated and compared, and some rules are found out: their maximal excitation wavelength and emission wavelength are in the range of 270-300 nm and 395-445 nm, respectively. And the PS-RTP characters of lifetime, polarization and quanta yield are also investigated and compared. It is found that their lifetimes of PS-RTP are all in the level of 0.1s. They belong to long life phosphorescence and their PS-RTP spectra are incompletely polarized. PMID- 16043052 TI - Synthesis and spectroscopic study on photochromism of a new thiosemicarbazone compound containing pyrazolone. AB - A new photochromic compound 1,3-diphenyl-4-(4'-fluro)benzal-5-pyrazolone-4-ethyl thiosemicarbazone (DP4FBP-ETSC) was synthesized by direct condensation of 1,3 diphenyl-5-pyrazolone with N4-ethyl thiosemicarbazide. The product was characterized by elemental analysis, IR and 1H NMR spectra. The photochromic properties of the compound were studied using time-dependent fluorescence emission spectra, the UV-vis reflection spectra in the solid state and the UV-vis absorption spectroscopy in liquid. The reaction rate constant of compound was also analyzed. The results show that DP4FBP-ETSC can perform photochromism. PMID- 16043053 TI - VO2+ ions in zinc lead borate glasses studied by EPR and optical absorption techniques. AB - Electron paramagnetic resonance (EPR) and optical absorption spectra of vanadyl ions in zinc lead borate (ZnO-PbO-B2O3) glass system have been studied. EPR spectra of all the glass samples exhibit resonance signals characteristic of VO2+ ions. The values of spin-Hamiltonian parameters indicate that the VO2+ ions in zinc lead borate glasses were present in octahedral sites with tetragonal compression and belong to C4V symmetry. The spin-Hamiltonian parameters g and A are found to be independent of V2O5 content and temperature but changing with ZnO content. The decrease in Deltag( parallel)/Deltag( perpendicular) value with increase in ZnO content indicates that the symmetry around VO2+ ions is more octahedral. The decrease in intensity of EPR signal above 10 mol% of V2O5 is attributed to a fall in the ratio of the number of V4+ ions (N4) to the number of V5+ ions (N5). The number of spins (N) participating in resonance was calculated as a function of temperature for VO2+ doped zinc lead borate glass sample and the activation energy was calculated. From the EPR data, the paramagnetic susceptibility was calculated at various temperatures and the Curie constant was evaluated from the 1/chi-T graph. The optical absorption spectra show single absorption band due to VO2+ ions in tetragonally distorted octahedral sites. PMID- 16043054 TI - Synthesis, spectra and biomimetic properties of copper(II)-copper(II) and copper(II)-zinc(II) binuclear complexes with CuN5 chromophores. AB - X-band electron spin resonance (ESR) and UV-vis spectra of a homobinuclear [(Bipy)2Cu-E-Im-Cu(Bipy)2](BF4)3 and a heterobinuclear [(Bipy)2Cu-E-Im Zn(Bipy)2](BF4)3 complexes, E-Im=2-ethylimidazolate ion have been described as possible models for superoxide dismutase (SOD). Magnetic moment and ESR spectral measurements of the homobinuclear complex have shown an antiferromagnetic exchange interaction. From pH-dependent ESR and UV-vis spectral measurements studies, these complexes have been found to be stable over 8.5-10.5 pH range. These complexes catalyze the dismutation of superoxide (O2-) at biological pH. All the observations indicate that these complexes act as good possible models for superoxide dismutase. PMID- 16043055 TI - Symmetric Euler orientation representations for orientational averaging. AB - A new kind of orientation representation called symmetric Euler orientation representation (SEOR) is presented. It is based on a combination of the conventional Euler orientation representations (Euler angles) and Hamilton's quaternions. The properties of the SEORs concerning orientational averaging are explored and compared to those of averaging schemes that are based on conventional Euler orientation representations. To that aim, the reflectance of a hypothetical polycrystalline material with orthorhombic crystal symmetry was calculated. The calculation was carried out according to the average refractive index theory (ARIT [T.G. Mayerhofer, Appl. Spectrosc. 56 (2002) 1194]). It is shown that the use of averaging schemes based on conventional Euler orientation representations leads to a dependence of the result from the specific Euler orientation representation that was utilized and from the initial position of the crystal. The latter problem can be overcome partly by the introduction of a weighing factor, but only for two-axes-type Euler orientation representations. In case of a numerical evaluation of the average, a residual difference remains also if a two-axes type Euler orientation representation is used despite of the utilization of a weighing factor. In contrast, this problem does not occur if a symmetric Euler orientation representation is used as a matter of principle, while the result of the averaging for both types of orientation representations converges with increasing number of orientations considered in the numerical evaluation. Additionally, the use of a weighing factor and/or non-equally spaced steps in the numerical evaluation of the average is not necessary. The symmetrical Euler orientation representations are therefore ideally suited for the use in orientational averaging procedures. PMID- 16043056 TI - Correlation of observed and model vibrational frequencies for aqueous organic acids: UV resonance Raman spectra and molecular orbital calculations of benzoic, salicylic, and phthalic acids. AB - The aromatic carboxylic acids benzoic, salicylic and phthalic acid were used to study the interaction of soluble organics compounds with metal cations. To accomplish this, we have developed methods for studying the carboxylic acids using UV resonance Raman (UVRR) combined with molecular orbital density functional theory calculations. The pH values of the acid solutions were based on the pK(a)'s for the different acids to examine the neutral and charged species. Deprotonation of the organic acids was detectable down to 10(-4)M using UVRR (two orders of magnitude lower than previous vibrational spectroscopy studies). Limitations to decreasing the concentration lower using the current UVRR facilities are discussed. Two methods were used to calculate the optimized geometry and frequencies of the acids: explicit and continuum solvation. The frequencies from the experimental spectra were then compared to the theoretical results obtained from the two methods. PMID- 16043057 TI - Changes in chemical composition and collagen structure of dentine tissue after erbium laser irradiation. AB - Erbium laser radiation has a great affinity for the water molecule, which is present in quantity in biological hard tissues. The objective of this work is to identify chemical changes by infrared spectroscopy of irradiated dentine by an Er:YAG-2.94 microm laser. The irradiation was performed with fluences between 0.365 and 1.94 J/cm2. For the infrared analysis a Fourier transform infrared spectrometer was used. After the irradiation were observed: loss of water, alteration of the structure and composition of the collagen, and increase of the OH- radical. These alterations can be identified by a decrease in intensity of the water band between 2800-3800 cm(-1), OH- band at 3575 cm(-1) and bands ascribed to organic matrix between 2800-3400 cm(-1) and 1100-1400 cm(-1). PMID- 16043058 TI - The study on photoreflectance characteristic of semi-insulating GaAs surface region by its exposure to 6 MeV electron beam. AB - Photoreflectance measurements were performed to investigate the optical properties in the electron beam irradiation semi-insulating GaAs(e-beam irradiation GaAs) and semi-insulating GaAs(SI-GaAs). A considerable increase of the PR amplitudes has been registered after the e-beam irradiation in comparison with the GaAs. It is that result of a higher electron scattering on the lattice defects created by the e-beam. PMID- 16043059 TI - Synthesis and spectroscopic behavior of highly luminescent Eu(3+) dibenzoylmethanate (DBM) complexes with sulfoxide ligands. AB - In this paper the synthesis, characterization and photoluminescent behavior of the [RE(DBM)3L2] complexes (RE=Gd and Eu) with a variety of sulfoxide ligands; L=benzyl sulfoxide (DBSO), methyl sulfoxide (DMSO), phenyl sulfoxide (DPSO) and p tolyl sulfoxide (PTSO) have been investigated in solid state. The emission spectra of the Eu(3+)-beta-diketonate complexes show characteristics narrow bands arising from the 5D0-->7F(J) (J=0-4) transitions, which are split according to the selection rule for C(n), C(nv) or C(s) site symmetries. The experimental Judd Ofelt intensity parameters (Omega2 and Omega4), radiative (A(rad)) and non radiative (A(nrad)) decay rates, and R02 for the europium complexes have been determined and compared. The highest value of Omega2 (61.9x10(-20)cm2) was obtained to the complex with PTSO ligand, indicating that Eu3+ ion is in the highly polarizable chemical environment. The higher values of the experimental quantum yield (q) and emission quantum efficiency of the emitter 5D0 level (eta) for the Eu-complexes with DMSO, DBSO and PTSO sulfoxides suggest that these complexes are promising Light Conversion Molecular Devices (LCMDs). The lower value of quantum yield (q=1%), for the hydrated complex [Eu(DBM)3H2O], indicates that the luminescence quenching occurs via multiphonon relaxation by coupling with the OH-oscillators from water molecule coordinated to rare earth ion. The pure red emission of the Eu-complexes has been confirmed by (x, y) color coordinates. PMID- 16043060 TI - Study of the interaction of proteins with curcumin and SDS and its analytical application. AB - It is found that protein and sodium dodecyl sulphonate (SDS) can enhance resonance light scattering (RLS) of curcumin (CU). Based on this phenomenon, a new quantitative method for protein in aqueous solution has been developed. In the BR (pH 3.5) buffer, the RLS intensity of CU-SDS system is greatly enhanced by protein. The enhanced RLS is proportional to the concentration of protein in the range of 0.00020-20.0 microgml(-1) for bovine serum albumin (BSA) and 0.00040-1.0 microgml(-1) for human serum albumin (HSA) and their detection limits are 0.16 and 0.041 ngml(-1), respectively. An actual sample is satisfactorily determined. In addition, the interaction mechanism between protein and CU-SDS is also studied by using multi-techniques such as RLS, absorption spectroscopy and fluorescence, zeta potential assay measurement. PMID- 16043061 TI - Qualitative study of ethanol content in tequilas by Raman spectroscopy and principal component analysis. AB - Using Raman spectroscopy, with an excitation radiation source of 514.5 nm, and principal component analysis (PCA) was elaborated a method to study qualitatively the ethanol content in tequila samples. This method is based in the OH region profile (water) of the Raman spectra. Also, this method, using the fluorescence background of the Raman spectra, can be used to distinguish silver tequila from aged tequilas. The first three PCs of the Raman spectra, that provide the 99% of the total variance of the data set, were used for the samples classification. The PCA1 and PCA2 are related with the water (or ethanol) content of the sample, whereas the PCA3 is related with the fluorescence background of the Raman spectra. PMID- 16043062 TI - Vibrational spectra of tris(dmit) complexes of main group metals: infrared, Raman and ab initio calculations. AB - Infrared Fourier Transform investigation of several metal tris-complexes of 1,3 dithiole-2-thione-4,5-dithiolate (dmit) ligand have been recorded within a theoretical-experimental investigation of the vibrational molecular spectra of crystalline [NEt4]2[Sn(dmit)3] and [NEt4][Sb(dmit)3] compounds. For the [Sn(dmit)3]-2 anion we recorded as well the Raman Fourier Transform spectra. Ab initio calculations have been carried with several ECPs, basis sets and methodologies (RHF and DFT) in order to assess family and methodological errors precisely. Geometry optimization and subsequent hessian calculation lead to the vibration frequencies reported. These calculated frequencies and intensities assisted the fundamental, overtones and combinations bands assignments. Remarkable agreement has been found between the experimental geometries and frequencies to those here calculated. Besides the bands traditionally studied for the dmit compounds, as CC and CS stretchings, also the region below 500 cm(-1) was evaluated, allowing to characterize several modes involving angular deformation of the dmit as the MS stretching of these octahedral distorted systems. PMID- 16043063 TI - Fluorescence study of Eu chelates in solution and polymer matrix. AB - The fluorescence spectra of Eu chelates with different neutral ligand in methyl methacryate (MMA) solution were measured and the influence of the neutral ligand was analyzed. The chelate Eu(DBM)3(TOPO)2, which shows the strongest fluorescence intensity in the solution, has been incorporated into the poly(methyl methacryate) (PMMA) matrix. According to the fluorescence emission spectrum, its radiative properties were also predicted. PMID- 16043064 TI - Application of probability circles analysis to the construction of calibration curves for infra red spectroscopy. AB - A new method, based on the presentation of initial data in the form of probability circles (PC), has been developed for constructing a calibration curve, which shows a monotonic dependence with respect to the given concentration. The centre of the reference probability circle is defined by its mean value and the radius of the circle is calculated as the value of the standard deviation of the sampling considered. The comparative probability circle is defined by the same corresponding parameters but rotated, relative to the initial reference circle, by an angle, which is related to the Pearson's correlation coefficient (PCC). The two parameters of the PCC and the statistical proximity factor (PCF), which defines the positions of the centres of the circles relative to each other, can be chosen as statistical parameters for the construction of the desired calibration curve. Experiments realized with the mixture of two liquids (chloroform serves as the basic matrix) and acetone (serves as an additive) confirm the efficiency of this new analytical method and demonstrate a possible increase sensitivity for the detection of lower concentration limit by approximately one order of magnitude. This new approach, which is free from model assumptions, and having very clear geometrical meaning, can be applied for different types of spectra and has many potential applications in the construction of calibration curves for different additives embedded within different matrices. PMID- 16043065 TI - Raman spectroscopic study of the hydrotalcite desautelsite Mg6Mn2CO3(OH)16 x 4H2O. AB - The structure of the hydrotalcite desautelsite Mg6Mn2CO3(OH)16.4H2O has been studied by a combination of Raman and infrared spectroscopy. Three intense Raman bands are observed at 1086, 1062 and 1055 cm(-1). A model based upon the observation of three CO3 stretching vibrations is presented. The CO3 anion may be (a) non-hydrogen bonded, (b) hydrogen bonded to the interlayer water and (c) hydrogen bonded to the brucite-like hydroxyl surface. Two intense bands at 3646 and 3608 cm(-1) are attributed to MgOH and MnOH stretching vibrations. Infrared bands at 3476, 3333, 3165 and 2991 cm(-1) are assigned to water stretching bands. Raman spectroscopy has proven a powerful tool for the study of hydrotalcite minerals. PMID- 16043066 TI - A Raman spectroscopic study of the uranyl sulphate mineral johannite. AB - Raman spectroscopy at 298 and 77K has been used to study the secondary uranyl mineral johannite of formula (Cu(UO2)2(SO4)2(OH)2 x 8H2O). Four Raman bands are observed at 3593, 3523, 3387 and 3234cm(-1) and four infrared bands at 3589, 3518, 3389 and 3205cm(-1). The first two bands are assigned to OH- units (hydroxyls) and the second two bands to water units. Estimations of the hydrogen bond distances for these four bands are 3.35, 2.92, 2.79 and 2.70 A. A sharp intense band at 1042 cm(-1) is attributed to the (SO4)2- symmetric stretching vibration and the three Raman bands at 1147, 1100 and 1090cm(-1) to the (SO4)2- anti-symmetric stretching vibrations. The nu2 bending modes were at 469, 425 and 388 cm(-1) at 77K confirming the reduction in symmetry of the (SO4)2- units. At 77K two bands at 811 and 786 cm(-1) are attributed to the nu1 symmetric stretching modes of the (UO2)2+ units suggesting the non-equivalence of the UO bonds in the (UO2)2+ units. The band at 786cm(-1), however, may be related to water molecules libration modes. In the 77K Raman spectrum, bands are observed at 306, 282, 231 and 210cm(-1) with other low intensity bands found at 191, 170 and 149cm(-1). The two bands at 282 and 210 cm(-1) are attributed to the doubly degenerate nu2 bending vibration of the (UO2)2+ units. Raman spectroscopy can contribute significant knowledge in the study of uranyl minerals because of better band separation with significantly narrower bands, avoiding the complex spectral profiles as observed with infrared spectroscopy. PMID- 16043067 TI - Spectroscopic studies on [(DD18C6H2)(HPA)2](PA)2 and [(DD18C6H2)(DDQ)2](DDQH)2 formed in the reaction of N,N'-dibenzyl-1,4,10,13-tetraoxa-7,16 diazacyclooctadecane with HPA and DDQ. AB - The interaction of the mixed oxygen-nitrogen cyclic base, N,N'-dibenzyl-1,4,10,13 tetraoxa-7,16-diazacyclooctadecane (DD18C6) with pi-acceptors such as picric acid (HPA) and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) has been studied spectrophotometrically in chloroform at 25 degrees C. The results obtained indicate the formation of 1:4 charge-transfer complexes with the general formula (DD18C6)(acceptor)4. The electronic and infrared spectra of charge-transfer complexes along with the (1)H NMR spectra were recorded and discussed. Based on the data obtained, the complexes were formulated as [(DD18C6H2)(HPA)2](PA)2 and [(DD18C6H2)(DDQ)2](DDQH)2. A general mechanism explaining the formation of the DDQ complex has been suggested. PMID- 16043068 TI - Structural and orientational studies of chiral N-[4(1-pyrene)butyroyl] phenylalanine in cast film. AB - Cast films of chiral material N-[4(1-pyrene)butyroyl]-l-phenylalanine (Py-l-Phe) and the racemic modification (the 1:1 mixture of Py-l-Phe and Py-d-Phe) have been investigated in the present study. The microscopic structure, molecular orientation and the aggregation in the film have been studied by using ultraviolet-visible (UV-vis) and infrared (IR) spectroscopy. When a cast film is formed from the chloroform solution at room temperature, the J-aggregates of Py-l Phe and Py-d-Phe are observed by UV-vis spectra. A comparison of Fourier transform infrared (FT-IR) transmission and reflection-absorption (RA) spectroscopy has been applied to reveal the orientation and structural characterization of the cast films. It has been found that the pyrenyl ring in the cast film of Py-l-Phe assumes a nearly vertical orientation with respect to the surface of the solid substrate. And there are two different hydrogen bonding species, cyclic dimer and linear dimer, exist in carboxyl groups in cast films. The detailed analysis of the OH and NH stretching modes of Py-l-Phe and Py-d-Phe allows us to reveal the hydrogen bonds existing in the films. PMID- 16043069 TI - EPR paramaters and defect structure of the trigonal Ti2+ center in ZnS. AB - The axial Ti2+ center in a nearly wholly cubic ZnS crystal is assigned to the Ti2+ ion on the hexagonal site of wurtzite structure caused by stacking faults. On the ground of the assignment, the EPR parameters (zero-field splitting D, g factor g( parallel) and g-anisotropy Deltag=g( parallel)-g( perpendicular)) of the axial Ti2+ center are calculated from the high-order perturbation formulas based on the cluster approach for the EPR parameters of 3d2 ion in trigonal symmetry. From the calculations, the local atom-position parameter u(loc) (which is different from the corresponding parameter u in the host wurtzite structure) and hence the defect structure of the Ti2+ center are estimated. The results (the calculated EPR parameters and the defect structure) are discussed. PMID- 16043070 TI - Growth and vibrational spectroscopic studies of pure and yttrium doped single crystals of trisodium barium pentachloride dihydrate. AB - Single crystals of pure and rare earth doped trisodium barium pentachloride dihydrate have been grown by slow evaporation solution growth. The solubility of both pure and doped forms of Na3BaCl5 x 2H2O has been estimated. Vibrational spectra were recorded to determine the symmetries of molecular vibrations. The observed Raman and infrared bands were assigned and discussed in detail. The optical transparencies of the grown crystals were tested by UV-vis spectrophotometer. PMID- 16043071 TI - Spectroscopic properties of gallium arsenide tetramers: Ga2As2, Ga2As2+ and Ga2As2-. AB - Spectroscopic properties of the low-lying electronic states of Ga2As2 and its ions are studied using the complete active-space self-consistent field (CASSCF) and density function theory (DFT) followed by the coupled-cluster single and double substitutions (including triple excitations) (CCSD(T)) calculations. The stability of low-lying electronic states is examined by computing vibrational frequency. The energies of the ground states and a number of excited electronic states have been computed to predict the spectra of Ga2As2 and its ions. The ionization energy, electronic affinity, and atomization energy are estimated at the CCSD(T)/6-311+G(d) level and compared with the available experimental results. PMID- 16043072 TI - Direct fluorimetric determination of ascorbic acid by the supramolecular system of AA with beta-cyclodextrin derivative. AB - We established a simple and selective fluorimetric method for the determination of ascorbic acid (AA) with the novel mono-[6-N(4-carboxy-phenyl)]-beta cyclodextrin (ACD). The method is based on the fluorescence intensity of ACD decreases as the ACD-AA supramolecular complexes form. The fluorescence intensity is measured at excitation and emission wavelengths of 270 and 352 nm, respectively. Under optimum condition, a linear relationship is obtained between the fluorescence intensity and the concentration of AA in the range of 0.05-8.0 microgml(-1). The detection limit is 0.012 microgml(-1). The method has been applied to the direct analysis of AA in real samples with satisfactory results. PMID- 16043073 TI - Vibrational spectrum of glycine molecule. AB - The infrared and Raman spectra of glycine molecule has been studied in spectral region 400-4000 cm(-1) in solid form as well as in water. The vibrational frequencies for the fundamental modes of the glycine in neutral and its zwitterionic form have also been calculated using AM1 semiempirical method as well as ab initio method with minimal basis set. The reliability of the minimal basis set and AM1 method with higher basis sets, for IR spectra of the neutral glycine conformers were examined. We find that the 6-21G basis set calculation yields structural parameters, rotational constant and dipole moment of glycine conformers, which are very similar to those obtained from extended basis set calculation as well as experimental values. IR frequencies for glycine conformer I are also calculated in water using SCRF=PCM model and compared with experimental values. A comparison between calculated frequencies for neutral glycine, and its zwitterionic form with observed IR and Raman bands have been made. The total energies for gas phase glycine and its zwitterionic form along with those of hydrated forms were also calculated. It is found from the calculations that in the gas phase neutral glycine is more stable as compared to its zwitterionic form. PMID- 16043074 TI - 2,4,5,7-Tetranitrofluorescein in solutions: novel type of tautomerism in hydroxyxanthene series as detected by various spectral methods. AB - Stepwise dissociation and tautomerism of 2,4,5,7-tetranitrofluorescein (TNF) were studied by using vis-spectroscopy in dimethylsulfoxide (DMSO), in aqueous acetone, and in cetyl-trimethylammonium chloride (CTAC) micellar solutions at ionic strength of the bulk phase 4.00M KCl. The pK(a) values in DMSO and 90 mass% (CH3)2CO as well as the 'apparent'pK(a)(a) values of the substance in micellar media were determined spectrophotometrically. The neutral (molecular) form H2R is found to be completely converted into the colorless lactone. Moreover, the lactonic structure, yellow due to 'nitrophenolate' absorption band, predominates also in the case of TNF dianion R2-. Contrary to the unsubstituted fluorescein, and like 2,4,5,7-tetrabromofluorescein (eosin), the monoanion HR- of TNF with lambda(max) 522-525 nm and E(max) approximately (60-62)x10(3) dm(3)mol(-1)cm(-1) exists mainly as a deeply and intensively colored structure with non-ionized carboxylic and ionized hydroxylic group; its fluorescence spectra in various media are registered. In 90% acetone, the Stokes shift is 1.17x10(3)cm(-1), fluorescence lifetime equals 2.3 ns. An extremely expressed trend to dianion lactone formation of R2- ion of TNF is confirmed in the systems studied. For TNF in DMSO, in aqueous acetone, in surfactant micelles, and in trichloromethane extracts of ionic associatiates with N(n-Bu)4+ and N(n-Hept)4+, the deeply colored 'quinon-phenolate' dianion, typical for all hydroxyxanthenes, is not registered at all. The sequence of dissociation of functional groups in solution is confirmed using IR spectroscopy in DMSO. PMID- 16043075 TI - Lanthanum(III) and praseodymium(III) complexes with isatin thiosemicarbazones. AB - Ten new lanthanum(III) and praseodymium(III) complexes of the general formula Na[La(L)2H2O] (Ln=La(III) or Pr(III); LH2=thiosemicarbazones) derived from the condensation of isatin with 4-phenyl thiosemicarbazide, 4-(4-chlorophenyl) thiosemicarbazide, 4-(2-nitrophenyl) thiosemicarbazide, 4-(2-bromophenyl) thiosemicarbazide and 4-(2-methylphenyl) thiosemicarbazide, have been synthesized in methanol in presence of sodium hydroxide. The XRD spectra of the complexes were monitored to verify complex formation. The complexes have also been characterized by elemental analysis, molar conductance, electronic absorption and fluorescence, infrared, far infrared, 1H and 13C NMR spectral studies. Thermal studies of these complexes have been carried out in the temperature range 25-800 degrees C using TG, DTG and DTA techniques. All these complexes decompose gradually with the formation of Ln2O3 as the end product. The Judd-ofelt intensity parameter, oscillator strength, transition probability, stimulated emission cross section for different transitions of Pr3+ for 4-phenyl thiosemicarbazones have been calculated. PMID- 16043076 TI - Vibrational frequencies and structural determination of dicyanodifluorosulfur. AB - The normal mode frequencies and corresponding vibrational assignments of dicyanodifluorosulfur are examined theoretically using the Gaussian03 set of quantum chemistry codes. Each of the vibrational modes was assigned to one of six types of motion predicted by a group theoretical analysis (CN stretch, SC stretch, SF stretch, FSC bend, SCN bend, and CSC bend) utilizing the C(2v) symmetry of the molecule. A set of uniform scaling factors was derived for each type of motion. Predicted infrared and Raman intensities are reported. PMID- 16043077 TI - Electronic structure and biological activity of nucleobases. AB - HeI and HeII photoelectron spectra of 6-chloro-1,3-dimethyluracil have been measured. The assignment of the spectrum was made by comparison with photoelectron spectra of related compounds and by high-level OVGF calculations. The electronic structure changes in substituted nucleobases are discussed on the basis of photoelectron spectroscopic data. The electronic structure data are compared with structure-activity relationships regarding enzyme inhibition and complex formation. The electronic structure data give some insight into the nature of binding between nucleobases' derivatives and different enzymes whose activity they inhibit. PMID- 16043078 TI - Crystal structure, differential scanning calorimetry and vibrational low temperature investigation of C(NH2)3 x HSeO4. AB - The first X-ray and vibrational spectroscopic analysis of a new molecular complex between guanidinium and selenic acid is reported. The crystal of guanidinium hydrogenselenate at room temperature belongs to P2(1)/n space group of the monoclinic system with Z=4, a=8.330A, b=5.109A, c=14.855A and beta=92.65 degrees . Room temperature powder infrared and Raman spectra for the titled complex (1:1) were measured. The observed IR and Raman spectra are in accordance with this crystallographic structure. The differential scanning calorimetric (DSC) experiment on powder samples indicates on continuous phase transition at ca. 160K. To explain in detail the behavior of the crystal during the phase transition the infrared and Raman powder spectra in low temperature range (10 300K) were measured. The temperature dependencies of bands position and intensities for obtained spectra are analysed. PMID- 16043079 TI - Synthesis, proton and 13C NMR and reaction mechanism studies of novel isoindolones derivatives, obtained through TAWERS procedure. AB - A series of novel isoindolone derivatives (13-18) were prepared in goods yields by applying the TAWERS methodology. A general approach for formation of isoindolones from the aza-Wittig reaction of iminophosphoranes with dialdehydes under neutral and mild conditions was realized. Using 1D NMR a [1,3]-hydride migration was detected. The assignment of the structures and conformation behavior of the derivatives was achieved using 1D and 2D NMR (NOESY, DEPT, HMQC, and HMBC). PMID- 16043080 TI - Comment on vibrational spectral investigation on xanthine and its derivatives theophylline, caffeine and theobromine. PMID- 16043081 TI - Technology induced error and usability: the relationship between usability problems and prescription errors when using a handheld application. AB - This paper describes an innovative approach to the evaluation of a handheld prescription writing application. Participants (10 physicians) were asked to perform a series of tasks involving entering prescriptions into the application from a medication list. The study procedure involved the collection of data consisting of transcripts of the subjects who were asked to "think aloud" while interacting with the prescription writing program to enter medications. All user interactions with the device were video and audio recorded. Analysis of the protocols was conducted in two phases: (1) usability problems were identified from coding of the transcripts and video data, (2) actual errors in entering prescription data were also identified. The results indicated that there were a variety of usability problems, with most related to interface design issues. In examining the relationship between usability problems and errors, it was found that certain types of usability problems were closely associated with the occurrence of specific types of errors in prescription of medications. Implications for identifying and predicting technology-induced error are discussed in the context of improving the safety of health care information systems. PMID- 16043082 TI - eLearning techniques supporting problem based learning in clinical simulation. AB - This paper details the results of the first phase of a project using eLearning to support students' learning within a simulated environment. The locus was a purpose built clinical simulation laboratory (CSL) where the School's philosophy of problem based learning (PBL) was challenged through lecturers using traditional teaching methods. THE SOLUTION: a student-centred, problem based approach to the acquisition of clinical skills that used high quality learning objects embedded within web pages, substituting for lecturers providing instruction and demonstration. This encouraged student nurses to explore, analyse and make decisions within the safety of a clinical simulation. Learning was facilitated through network communications and reflection on video performances of self and others. Evaluations were positive, students demonstrating increased satisfaction with PBL, improved performance in exams, and increased self-efficacy in the performance of nursing activities. These results indicate that eLearning techniques can help students acquire clinical skills in the safety of a simulated environment within the context of a problem based learning curriculum. PMID- 16043083 TI - Understanding technology adoption in clinical care: clinician adoption behavior of a point-of-care reminder system. AB - BACKGROUND: Evaluation studies of clinical decision support systems (CDSS) have tended to focus on assessments of system quality and clinical performance in a laboratory setting. Relatively few studies have used field trials to determine if CDSS are likely to be used in routine clinical settings and whether reminders generated are likely to be acted upon by end-users. Moreover, such studies when performed tend not to identify distinct user groups, nor to classify user feedback. AIM: To assess medical residents' acceptance and adoption of a clinical reminder system for chronic disease and preventive care management and to use expressed preferences for system attributes and functionality as a basis for system re-engineering. DESIGN OF STUDY: Longitudinal, correlational study using a novel developmental trajectory analysis (DTA) statistical method, followed by a qualitative analysis based on user satisfaction surveys and field interviews. SETTING: An ambulatory primary care clinic of an urban teaching hospital offering comprehensive healthcare services. 41 medical residents used a CDSS over 10 months in their daily practice. Use of this system was strongly recommended but not mandatory. METHODS: A group-based, semi-parametric statistical modeling method to identify distinct groups, with distinct usage trajectories, followed by qualitative instruments of usability and satisfaction surveys and structured interviews to validate insights derived from usage trajectories. RESULTS: Quantitative analysis delineates three types of user adoption behavior: "light", "moderate" and "heavy" usage. Qualitative analysis reveals that clinicians of distinct types tend to exhibit views of the system consistent with their demonstrated adoption behavior. Drawbacks in the design of the CDSS identified by users of all types (in different ways) motivate a redesign based on current physician workflows. CONCLUSION: We conclude that this mixed methodology has considerable promise to provide new insights into system usability and adoption issues that may benefit clinical decision support systems as well as information systems more generally. PMID- 16043084 TI - Architectures and tools for innovative Health Information Systems: the Guide Project. AB - This paper describes the architecture of the Guide Project, a proposal for innovation of Health Information Systems, putting together medical and organizational issues through the Separation of Concerns paradigm. In particular, we focus on one building block of the architecture: the Guideline Management System handling the whole life cycle of computerized Clinical Practice Guidelines. The communication between the Guideline Management System and the other components of the project architecture is message-based, according to specific contracts that allow an easy integration of the components developed by different parties and, in particular, with legacy systems (i.e. existing electronic patient records). In turn, the Guideline Management System components are organized in a distributed architecture: an editor to formalize guidelines, a repository to store and publish them, an enactment system to implement guidelines instances in a multi-user environment and a reporting system able to completely trace any individual physician's guideline-based decision process. The repository is organized in different levels that can be international, national, regional, down to the specific health care organization, according to the healthcare delivery policy of a country. Different organizations can get Clinical Practice Guidelines from the repository, adapt and introduce them in clinical practice. PMID- 16043085 TI - Using OrgAhead, a computational modeling program, to improve patient care unit safety and quality outcomes. AB - As part of ongoing research to investigate the impact of patient characteristics, organization characteristics and patient unit characteristics on safety and quality outcomes, we used a computational modeling program, OrgAhead, to model patient care units' achievement of patient safety (medication errors and falls) and quality outcomes. We tuned OrgAhead using data we collected from 32 units in 12 hospitals in Arizona. Validation studies demonstrated acceptable levels of correspondence between actual and virtual patient units. In this paper, we report how we used OrgAhead to develop testable hypotheses about the kinds of innovations that nurse managers might realistically implement on their patient care units to improve quality and safety outcomes. Our focus was on unit-level innovations that are likely to be easier for managers to implement. For all but the highest performing unit (for which we encountered a ceiling effect), we were able to generate practical strategies that improved performance of the virtual units that could be implemented by actual units to improve safety and quality outcomes. Nurse managers have responded enthusiastically to the additional decision support for quality improvement. PMID- 16043086 TI - ISO reference terminology models for nursing: applicability for natural language processing of nursing narratives. AB - Natural language processing (NLP) systems have demonstrated utility in parsing narrative texts for purposes such as surveillance and decision support. However, there has been little work related to NLP of nursing narratives. The purpose of this study was to compare the semantic categories of a NLP system (Medical Language Extraction and Encoding [MedLEE] system) with the semantic domains, categories, and attributes of the International Standards Organization (ISO) reference terminology models for nursing diagnoses and nursing actions. All but two MedLEE diagnosis and procedure-related semantic categories mapped to ISO models. In some instances, we found exact correspondence between the semantic structures of MedLEE and the ISO models. In other situations (e.g. aspects of Site or Location), the ISO model was not as granular as MedLEE. For clinical procedure and non-invasive examination, two ISO nursing action model components (Action and Target) mapped to a single MedLEE semantic category. The ISO models are applicable to NLP of nursing narratives. However, the ISO models require additional specification of selected semantic categories for the abstract semantic domains in order to achieve the objective of using NLP to parse and encode data from nursing narratives. Our analysis also suggests areas for extension of MedLEE particularly in regard to represent nursing actions. PMID- 16043087 TI - Impact of CPOE on doctor-nurse cooperation for the medication ordering and administration process. AB - The objective of this study was to analyze the impact of medication ordering and administration functions of CPOE on doctor-nurse communications and cooperation. We performed an extensive analysis of the work situations in several departments of three different hospitals. One of the hospitals is still using paper-based orders, the second one is currently implementing a patient care information system (PCIS), but the analysis was carried out with paper-based orders. The third hospital has a PCIS installed with available medication ordering functions. We used standard methods from cognitive psychology to analyze physicians, and nurses' activity, communications and cooperation. This approach was combined with a usability analysis of both work systems, paper and computer-based. The paper based situation is characterized by a synchronous cooperation with a distributed decision-making where physicians and nurses rely mostly on verbal communications to coordinate their actions; paper order sheets are weakly structured and poorly support the documentation task. In the computer situation, physicians and nurses work in an asynchronous mode, and leave to the system the coordination of their actions. Orders are exhaustively documented but some data may be misinterpreted. Some of these problems are due to usability flaws of the Human Computer Interface. We conclude with recommendations for usability improvement of CPOE systems, combined with recommendations for the organization of doctor-nurse communication when implementing such systems. PMID- 16043088 TI - Differing faculty and housestaff acceptance of an electronic health record. AB - In order to determine whether differences exist between housestaff and faculty physician acceptance of an electronic health record system, we conducted a written survey of attitudes towards new electronic medical record at the University of Illinois at Chicago. We surveyed 330 faculty and housestaff physicians. User acceptance of the EHR was high for both faculty physicians and housestaff. An amount of 88.0% of the housestaff and 64.7% of the faculty preferred the EHR over a paper record. Although both housestaff and faculty acceptance of an EHR was high, housestaff showed greater approval ratings than faculty. Central to acceptance of an EHR is conservation of physician time, including improving system speed, reducing time spent waiting for a computer to become available, and minimizing time spent documenting care. PMID- 16043089 TI - Automatic generation of repeated patient information for tailoring clinical notes. AB - Generating clear, readable, and accurate reports can be a time-consuming task for physicians. Clinical notes, which document patient encounters, often contain a certain set of patient information including demographics, medical history, surgical history, examination results or the current medical condition that is propagated from one clinical note to all subsequent clinical notes for the same patient. To this end, we present a system, which automatically generates this patient information for the creation of a new clinical note. We use semantic patterns and an approximate sequence matching algorithm for capturing the discourse role of sentences, which we show to be a useful feature for determining whether the sentence should be repeated. Our system is shown to perform better than a simple baseline metric using precision/recall results. We believe such a system would allow clinical notes to be more complete, timely, and accurate. PMID- 16043090 TI - Usability of quality measures for online health information: Can commonly used technical quality criteria be reliably assessed? AB - PURPOSE: Many criteria have been developed to rate the quality of online health information. To effectively evaluate quality, consumers must use quality criteria that can be reliably assessed. However, few instruments have been validated for inter-rater agreement. Therefore, we assessed the degree to which two raters could reliably assess 22 popularly cited quality criteria on a sample of 42 complementary and alternative medicine Web sites. METHODS: We determined the degree of inter-rater agreement by calculating the percentage agreement, Cohen's kappa, and prevalence- and bias-adjusted kappa (PABAK). RESULTS: Our un calibrated analysis showed poor inter-rater agreement on eight of the 22 quality criteria. Therefore, we created operational definitions for each of the criteria, decreased the number of assessment choices and defined where to look for the information. As a result 18 of the 22 quality criteria were reliably assessed (inter-rater agreement > or = 0.6). CONCLUSIONS: We conclude that even with precise definitions, some commonly used quality criteria cannot be reliably assessed. However, inter-rater agreement can be improved with precise operational definitions. PMID- 16043091 TI - New frontiers for nursing and health care informatics. AB - Health care and health information have been around since the time of Hippocrates or even before. Through the historical evolution, it is observed that the knowledge and information that were simple and easy to learn and retain by that time, became much more complex. This paper presents a brief reviewing on the evolution of MEDINFO conferences, and how nursing informatics grew up and made itself visible during all these years of IMIA conferences. PMID- 16043092 TI - Biological effects of rAAV-caAlk2 coating on structural allograft healing. AB - Structural bone allografts often fracture due to their lack of osteogenic and remodeling potential. To overcome these limitations, we utilized allografts coated with recombinant adeno-associated virus (rAAV) that mediate in vivo gene transfer. Using beta-galactosidase as a reporter gene, we show that 4-mm murine femoral allografts coated with rAAV-LacZ are capable of transducing adjacent inflammatory cells and osteoblasts in the fracture callus following transplantation. While this LacZ vector had no effect on allograft healing, bone morphogenetic protein signals delivered via rAAV-caAlk2 coating induced endochondral bone formation directly on the cortical surface of the allograft by day 14. By day 28 there was evidence of remodeling of the new woven bone and massive osteoclastic resorption of the cortical surface of the rAAV-caAlk2-coated allografts only. Micro-CT analysis of rAAV-LacZ- vs rAAV-caAlk2-coated allografts after 42 days of healing demonstrated a significant increase in new bone formation (0.67 +/- 0.21 vs 2.49 +/- 0.40 mm(3); P < 0.005). Furthermore, the 3D micro-CT images of femurs grafted with rAAV-Alk2-coated allografts provided the first evidence that complete bridging of bone around a cortical allograft is possible. These results indicate that cell-free, rAAV-coated allografts have the potential to revitalize in vivo following transplantation. PMID- 16043093 TI - Gene-induced chondrogenesis of primary mesenchymal stem cells in vitro. AB - Adult mesenchymal stem cells (MSCs) have the capacity to differentiate into various connective tissues such as cartilage and bone following stimulation with certain growth factors. However, less is known about the capacity of these cells to undergo chondrogenesis when these proteins are delivered via gene transfer. In this study, we investigated chondrogenesis of primary, bone marrow-derived MSCs in aggregate cultures following genetic modification with adenoviral vectors encoding chondrogenic growth factors. We found that adenoviral-mediated expression of TGF-beta1 and BMP-2, but not IGF-1, induced chondrogenesis of MSCs as evidenced by toluidine blue metachromasia and immunohistochemical detection of type II collagen. Chondrogenesis correlated with the level and duration of expressed protein and was strongest in aggregates expressing 10-100 ng/ml transgene product. Transgene expression in all aggregates was highly transient, showing a marked decrease after 7 days. Chondrogenesis was inhibited in aggregates modified to express >100 ng/ml TGF-beta1 or BMP-2; however, this was found to be partly due to the inhibitory effect of exposure to high adenoviral loads. Our findings indicate that parameters such as these are important functional considerations for adapting gene transfer technologies to induce chondrogenesis of MSCs. PMID- 16043094 TI - Improved tissue repair in articular cartilage defects in vivo by rAAV-mediated overexpression of human fibroblast growth factor 2. AB - Therapeutic gene transfer into articular cartilage is a potential means to stimulate reparative activities in tissue lesions. We previously demonstrated that direct application of recombinant adeno-associated virus (rAAV) vectors to articular chondrocytes in their native matrix in situ as well as sites of tissue damage allowed for efficient and sustained reporter gene expression. Here we test the hypothesis that rAAV-mediated overexpression of fibroblast growth factor 2 (FGF-2), one candidate for enhancing the repair of cartilage lesions, would lead to the production of a biologically active factor that would facilitate the healing of articular cartilage defects. In vitro, FGF-2 production from an rAAV delivered transgene was sufficient to stimulate chondrocyte proliferation over a prolonged period of time. In vivo, application of the therapeutic vector significantly improved the overall repair, filling, architecture, and cell morphology of osteochondral defects in rabbit knee joints. Differences in matrix synthesis were also observed, although not to the point of statistical significance. This process may further benefit from cosupplementation with other factors. These results provide a basis for rAAV application to sites of articular cartilage damage to deliver agents that promote tissue repair. PMID- 16043095 TI - Noggin improves bone healing elicited by muscle stem cells expressing inducible BMP4. AB - The aim of this study was to test the hypothesis that a specific antagonist may enhance the efficacy of its corresponding growth factor in a regulated tissue engineering strategy. Our prior research has led to the development of a retroviral vector that enables optimal regulated bone morphogenetic protein 4 (BMP4) expression in vitro and regulated bone formation in vivo with transduced muscle stem cells. However, when implanted in critical-sized calvarial defects, these cells led to residual bone formation without induction or bone overgrowth with induction, even at reduced cell doses. We thus co-implanted the aforementioned cells with stem cells engineered to express Noggin, a specific BMP antagonist. This approach, while preserving our ability to regulate bone regeneration closely, prevented both the basal level bone regeneration and the bone overgrowth and, more importantly, led to the regeneration of bone that more closely resembled normal bone. We believe that this regulatable tissue engineering strategy, enhanced by utilizing a specific antagonist, constitutes a new paradigm for tissue engineering and regenerative medicine. PMID- 16043096 TI - Gene transfer of the Runx2 transcription factor enhances osteogenic activity of bone marrow stromal cells in vitro and in vivo. AB - Marrow stromal cells (MSCs) have the potential to differentiate into multiple mesenchymal cell types. To harness the power of MSCs for bone regeneration, methods must be developed to direct their differentiation selectively to the osteoblast lineage. The objective of this study was to examine the feasibility of using ex vivo Runx2 gene transfer to enhance the osteogenic activity of MSCs. Primary MSCs isolated from C57BL6 mice were transduced with adenoviral vectors encoding beta-galactosidase or Runx2. Cells transduced with Ad-Runx2 expressed Runx2 protein and underwent osteoblast differentiation as measured by increases in alkaline phosphatase activity and mineralization. Time-course studies revealed that Runx2 protein was highest 1 day after transduction and declined below the limits of detection by 15 days. Osteoblast marker mRNA expression paralleled Runx2 levels. In contrast, Runx2-dependent mineralization persisted for the duration of the experiment. To assess in vivo osteogenic activity, Ad-Runx2 transduced and control MSCs were adsorbed to two different carrier scaffolds and subcutaneously implanted into C57BL6 mice. In both cases, MSCs expressing Runx2 formed substantially more bone than cells transduced with control virus. Taken together, these studies indicate that Runx2 gene transfer may be an effective route to enhance the osteogenic potential of MSCs. PMID- 16043097 TI - Characterization of hemodynamic events following intravascular infusion of recombinant adenovirus reveals possible solutions for mitigating cardiovascular responses. AB - Intravascular administration of recombinant adenovirus (rAd) in cancer patients has been well tolerated. However, dose-limiting hemodynamic responses associated with suppression of cardiac output have been observed at doses of 7.5 x 10(13) particles. While analysis of hemodynamic responses induced by small-molecule pharmaceuticals is well established, little is known about the cardiovascular effects of rAd. Telemetric cardiovascular (CV) monitoring in mice was utilized to measure hemodynamic events following intravascular rAd administration. Electrocardiogram analysis revealed a block in the SA node 3-4 min postinfusion, resulting in secondary pacemaking initiated at the AV node. This was associated with acute bradycardia, reduced blood pressure, and hypothermia followed by gradual recovery. Adenovirus-primed murine sera with high neutralizing antibody (nAb) titers could inhibit CV responses, whereas human sera with equivalent nAb titers induced by natural infection were, surprisingly, not inhibitory. Interestingly, repeat dosing within 2-4 h of the primary injection resulted in desensitization, resembling tachyphylaxis, for subsequent CV responses. Last, depletion of Kupffer cells prior to rAd infusion precluded induction of CV responses. These inhibitory effects suggest that rAd interactions with certain cells of the reticular endothelial system are associated with induction of CV responses. Significantly, these studies may provide insight into management of acute adverse effects following rAd systemic delivery, enabling a broadening of therapeutic index. PMID- 16043098 TI - Ex vivo transduction of human dermal tissue structures for autologous implantation production and delivery of therapeutic proteins. AB - Systemic delivery of therapeutic proteins through gene transfer approaches has been carried out mostly by ex vivo transduction of single cells or by direct in vivo injection of an expression vector. In this work an intact miniature biopsy of human dermis (microdermis) is harvested and transduced ex vivo by a viral vector encoding a gene for the therapeutic protein. The microdermis preserves its three-dimensional structure and viability during the ex vivo manipulations. Furthermore, upon transduction with adenoviral and adeno-associated viral vectors the microdermis secretes recombinant human erythropoietin (hEPO). Biochemical analysis of the secreted hEPO showed similarity to the clinically approved recombinant hEPO. Subcutaneous implantation of microdermal hEPO into SCID mice exhibited hEPO secretion in the blood circulation and preserved elevated hematocrit for several months, demonstrating the technology's potential for sustained delivery of protein therapeutics. PMID- 16043099 TI - No evidence for tumorigenesis of AAV vectors in a large-scale study in mice. AB - Six hundred ninety-five mice received adeno-associated virus (AAV) vectors, mostly via portal vein injection. At necropsy, the livers were inspected for tumors, and tissue sections were prepared for histology. We observed only one tumor, a lipoma, resulting in a tumor frequency of 0.14%. This tumor contained fewer vector genomes per total DNA than the surrounding liver tissue, as shown by quantitative PCR. In another mouse we found a macroscopically visible nodule containing lymphocytes. Immunohistochemistry revealed cells not of monoclonal origin, and they contained fewer AAV genomes than the surrounding hepatocytes. There were no macroscopic tumors in 226 control mice. Upon microscopic examination, lymphocytic infiltrates were found in 5% of livers of both control and vector-treated mice; no transgene expression was seen in those infiltrates in AAV-injected animals. Compared to an average frequency of spontaneous liver tumors in C57BL/6 mice (0-10%), and given the absence of high levels of vector DNA in the observed tumor, we conclude that AAV vectors do not predispose these target animals to the formation of liver tumors. PMID- 16043100 TI - Targeting endothelial cells with adenovirus expressing nitric oxide synthase prevents elevation of blood pressure in stroke-prone spontaneously hypertensive rats. AB - Local adenoviral (Ad)-mediated gene transfer to the carotid artery of the stroke prone spontaneously hypertensive rat (SHRSP) is successful in improving endothelial function. Here we explored the potential of systemic delivery of Ad encoding endothelial nitric oxide synthase (AdeNOS) to prevent elevation of blood pressure in the SHRSP using both nontargeted and vector targeting approaches. Systemic administration of nontargeted AdeNOS failed to modify the rise in blood pressure in SHRSP when administered during the 12th week of age (n = 5, P = 0.088, F = 3.0), an effect likely to result from sequestration of Ad by the liver. Rerouting Ad transduction using a bispecific antibody (anti-ACE/anti-Ad capsid, Fab9B9) that blocks Ad binding to the coxsackie and adenovirus receptor and simultaneously retargets AdeNOS to the angiotensin-converting enzyme resulted in efficient eNOS overexpression in the lung vasculature and a sustained hypotensive effect (n = 5, P = 0.007, F = 7.9). This study highlights the importance of vector targeting to achieve therapeutic gain and represents the first such study in cardiovascular gene therapy. PMID- 16043101 TI - Adenoviral-mediated delivery of early growth response factor-1 gene increases tissue perfusion in a murine model of hindlimb ischemia. AB - To test the hypothesis that overexpression of early growth response factor-1 (Egr 1) contributes to the revascularization of ischemic limbs, a constitutively active form of Egr-1 (Egr-1*) was made and evaluated in vitro and in vivo. Analyses of the transduced myocytes revealed significant upregulation of bFGF, PDGF-A, PDGF-B, IGF-II, and TGF-beta1. A coculture assay of the paracrine effects indicated that Ad-Egr-1* promoted proliferation and migration of endothelial cells. When Ad-Egr-1* was injected into the tibialis anterior muscle of mice, followed by explant culture in growth factor-reduced Matrigel, many capillary like structures were observed in the Egr-1* group compared with minimal sprouting from the LacZ group, suggesting an angiogenic potential of Egr-1*. Next we evaluated Ad-Egr-1* in a murine model of hindlimb ischemia. Compared with slow revascularization in the control PBS or LacZ group, a rapid increase in tissue perfusion was observed in the Egr-1* group and the difference in flux ratio was statistically significant at day 7. In the injected muscle, expression of Egr-1*, upregulation of its target genes, and increased number of vessels staining positive for smooth muscle alpha-actin were observed. These results suggest that Egr-1 plays an important role in vascular recovery after occlusion and could be a potential target for therapeutic angiogenesis. PMID- 16043102 TI - Low therapeutic threshold for hepatocyte replacement in murine phenylketonuria. AB - Phenylalanine homeostasis in mammals is primarily controlled by liver phenylalanine hydroxylase (PAH) activity. Inherited PAH deficiency (phenylketonuria or PKU) leads to hyperphenylalaninemia in both mice and humans. A low level of residual liver PAH activity ensures near-normal dietary protein tolerance with normal serum phenylalanine level, but the precise threshold for normal phenylalanine clearance is unknown. We employed hepatocyte transplantation under selective growth conditions to investigate the minimal number of PAH expressing hepatocytes necessary to prevent hyperphenylalaninemia in mice. Serum phenylalanine levels remained normal in mice exhibiting nearly complete liver repopulation with PAH-deficient hepatocytes (<5% residual wild-type liver PAH activity). Conversely, transplantation of PAH-positive hepatocytes into PAH deficient Pah(enu2) mice, a model of human PKU, yielded a significant decrease in serum phenylalanine (<700 muM) when liver repopulation exceeded approximately 5%. These data suggest that restoration of phenylalanine homeostasis requires PAH activity in only a minority of hepatocytes. PMID- 16043103 TI - Defining the pathway for Tat-mediated delivery of beta-glucuronidase in cultured cells and MPS VII mice. AB - We used recombinant forms of human beta-glucuronidase (GUS) purified from secretions from stably transfected CHO cells to compare the native enzyme to a GUS-Tat C-terminal fusion protein containing the 11-amino-acid HIV Tat protein transduction domain for: (1) susceptibility to endocytosis by cultured cells, (2) rate of clearance following intravenous infusion, and (3) tissue distribution and effectiveness in clearing lysosomal storage following infusion in the MPS VII mouse. We found: (1) Native GUS was more efficiently taken up by cultured human fibroblasts and its endocytosis was exclusively mediated by the M6P receptor. The GUS-Tat fusion protein showed only 30-50% as much M6P-receptor-mediated uptake, but also was taken up by adsorptive endocytosis through binding of the positively charged Tat peptide to cell surface proteoglycans. (2) GUS-Tat was less rapidly cleared from the circulation in the rat (t(1/2) = 13 min vs 7 min). (3) Delivery to most tissues of the MPS VII mouse was similar, but GUS-Tat was more efficiently delivered to kidney. Histology showed that GUS-Tat more efficiently reduced storage in renal tubules, retina, and bone. These studies demonstrate that Tat modification can extend the range of tissues corrected by infused enzyme. PMID- 16043104 TI - Viral IL-10-mediated immune regulation in pancreatic islet transplantation. AB - Protection of transplanted pancreatic islet grafts in recipients with autoimmune diabetes depends on the suppression of autoimmune recurrence and allogeneic rejection. The aim of this study was to investigate the efficiency of viral IL-10 gene delivery in the prevention of autoimmune recurrence following islet transplantation. We evaluated the effectiveness of a systemically delivered adeno associated viral vector (AAV vIL-10) carrying viral IL-10 in protecting islet engraftment. We observed significant prolongation of graft survival after treatment with AAV vIL-10 when using islets from donors lacking autoimmunity. We found that the mechanism of vIL-10-mediated protection was associated with suppression of T cell activation and that donor immune cells that were simultaneously transferred with the islet grafts could induce autoimmune recurrence. AAV vIL-10 gene transfer suppressed previously activated T cells and protected grafted islets from autoimmune-mediated destruction. We conclude that vIL-10 can regulate autoimmune activity and that transfer of its gene may have potential for therapeutic islet transplantation. PMID- 16043105 TI - Local administration of an adeno-associated viral vector expressing IL-10 reduces monocyte infiltration and subsequent photoreceptor damage during experimental autoimmune uveitis. AB - Autoimmune posterior uveitis is a chronic, potentially blinding inflammatory disease of the eye. It is commonly treated with immunosuppressive drugs that have adverse long-term effects. Advances in gene transfer techniques have enabled long term, stable transduction of retinal cells following subretinal injection with adeno-associated viral (AAV) vectors. Here we report for the first time that subretinal injection of rAAV-2 encoding murine IL-10 into the retina of C57BL/6 mice significantly decreases the median experimental autoimmune uveitis (EAU) disease severity. This protection is shown to be due to a decrease in the number and activation status of infiltrating monocytes during EAU, as determined by costimulatory molecule expression and nitrotyrosine detection. No differences within splenocyte proliferative responses or serum antibody levels were detected, emphasizing the potential of gene therapy strategies in ameliorating autoimmune responses in local microenvironments without unwanted systemic effects. PMID- 16043106 TI - Road mobility and the risk of road traffic accident as a driver. The impact of medical conditions and life events. AB - STUDY OBJECTIVE: We conducted a longitudinal investigation of the impact of self reported life events and medical conditions on changes in road mobility on the wheel between 2000 and 2002 in order to assess whether these changes would affect the risk of road traffic accident (RTA). METHODS: Data are from a cohort of workers and retirees from the French national gas and electricity companies (the Gazel cohort). In the present study, 10,483 participants were included (7843 men aged 51-61 years and 2640 women aged 46-61 years, in 2000). The link between mobility and the risk of RTA was approximated using data on RTA number during lifetime and reported mobility in 2000. We then compared changes in road mobility between 2000 and 2002 resulting from life events and medical conditions reported to have occurred in the year 2001 or changed when compared to year 2000. We also compared road mobilities in 2000 in order to assess any pre-existing differences before life events and medical conditions. This led to estimation of the effect of road mobility changes on the risk of RTA. RESULTS: Changes in road mobility associated with life events and medical conditions were only found among men. These changes in road mobility were minimal. Ensuing changes in the risk of RTA were estimated to be small (odds-ratios ranged from 0.94 to 1.01). The only life events found to be associated with increased road mobility was an important purchase. Hospitalization, serious RTA, and retiring were associated with reduced road mobility. Concerning medical conditions, men who reported cataract, angina pectoris, diabetes, anxiety and stress, sleep disorder, and depression decreased their road mobility. CONCLUSION: We found no or moderate changes in road mobility resulting from life events and medical conditions, suggesting that results from previous published studies that assessed the impact of life events or medical conditions on RTA were not jeopardized by improper adjustment for road mobility. PMID- 16043107 TI - Work-related traffic crashes: a record linkage study. AB - AIM: To examine the characteristics of work-related traffic crashes involving drivers in New South Wales (NSW), Australia. METHODS: Probabilistic data record linkage were used to merge police crash records and workers compensation data for the period 1998-2002. RESULTS: The record linkage identified 13,124 drivers who were injured or died as a result of work-related traffic crash in New South Wales over the 5-year period. Approximately three quarters of driver casualties occurred during commuting (74.8%) with the rest occurring in the course of work. Male drivers made up around three quarters of these crashes and 93% of those that resulted in a fatality. Transport workers were the most frequent victims of work related crashes while on duty (20.8%), with drivers of heavy trucks representing about half (48%) of all fatalities resulting from on duty work-related crashes. Nearly 1 in 6 male drivers were speeding at the time of the crash (15%, 95% CI 14.2-15.7) compared to less than 1 in 10 female drivers (9%, 95% CI 8.3-9.8) of female drivers. Male drivers were also significantly more likely to be fatigued at the time of the crash 7.6% (95% CI 7.0-8.2) compared to females 4.2% (95% CI 3.7-4.8). No significant difference was observed in the proportion of crashes involving fatigue between on duty and commuting traffic crashes. CONCLUSIONS: The study demonstrates the value of record linkage techniques in addressing some of the limitations of work-related data systems and in providing a more complete picture of the circumstances of occupational road crashes. PMID- 16043108 TI - Responses to comments by Ishag Adam and Mustafa Idriss El Bechir on our paper "Risk factors for malaria infection and anemia for pregnant women in the Sahel area of Bandiagara, Mali" published in Acta Tropica 89 (1) 17-23. PMID- 16043109 TI - The role of interactions in defining the structure of mixed protein-surfactant interfaces. AB - The behaviour of dispersed systems is often controlled by the composition of the interface between the different phases. Because of the inherent differences between proteins and surfactants, when both are present at the interface complex pseudo two-dimensional structures can be formed. This review brings together work that allows these structures to be explained in terms of the interactions involved. The significance of both the interactions and the structures is discussed in terms of the effect on bulk properties in colloidal systems. PMID- 16043110 TI - Measuring long association phases using Biacore. PMID- 16043111 TI - Novel type of general protein assay using a chromogenic and fluorogenic amine reactive probe. AB - We report on a new and simple one-reagent method for general protein assay. It makes use of one of two new reactive labeling reagents presented here (and referred to as pyrylium [Py] labels). These can be applied for both photometric and fluorometric protein assays at near neutral pHs at room temperature. The Py labels undergo a large spectral change on conjugation to the amino group of proteins and typically change their color from blue to red. Therefore, and unlike in other assays, there is no need to separate the unconjugated (blue) label from the red conjugate, which can be determined by direct photometry with a limit of detection of 1.2 microg/ml for human serum albumin. The assay can be extended to fluorometry because the fluorescence of the free Py label is weak (with a quantum yield of <1%) but increases strongly (to >40%) on conjugation. The strong fluorescence of the red conjugates can be determined directly and without interference by the blue (and weakly fluorescent) free label. The fluorometric assay resulted in a limit of detection of 60 ng/ml for bovine serum albumin (BSA). Validation of the fluorescence assay of blood plasma samples spiked with BSA gave recoveries in the range from 91 to 103%. PMID- 16043112 TI - A homogeneous assay for relative affinity of binding proteins using a green fluorescent protein tag and membrane disk. AB - When the association between a ligand immobilized on a membrane disk and a fluorescence-labeled analyte was monitored with a fluorescent microplate reader, the time-dependent increase in fluorescence intensity of the reaction mixture was observed. A novel assay system for the specific interaction based on this phenomenon was designated the homogeneous assay for fluorescence concentrated on membrane (HAFCOM). In this study, streptococcal protein G (SpG) and glycogen binding subunit R5 of protein phosphatase 1 (PPP1R5) tagged by green fluorescent protein (GFP) were used as the fluorescence-labeled analytes, and the affinity change caused by various amino acid substitutions was measured with HAFCOM. From the site-directed mutagenesis of SpG and PPP1R5, it was clarified that (i) the association rate constant of the Lys454Pro/Glu456Gln mutant of SpG to goat immunoglobulin G was almost equivalent to that of the wild-type but its dissociation rate constant was about 2.7 times that of the wild-type and (ii) the amino acid substitutions of Phe180 in PPP1R5 reduced glycogen-binding by 30-50%. Since HAFCOM using the GFP-tagged analyte requires no special chemicals and instruments, this system can easily and economically assay the specific interaction between target protein and ligand. PMID- 16043113 TI - Detecting proton flux across chromatophores driven by F0F1-ATPase using N (fluorescein-5-thiocarbamoyl)-1,2-dihexadecanoyl-sn-glycero-3 phosphoethanolamine, triethylammonium salt. AB - N-(Fluorescein-5-thiocarbamoyl)-1,2-dihexadecanoyl-sn-glycero-3 phosphoethanolamine, triethylammonium salt (F-DHPE) is a lipid fluorescence dye sensitive to pH changes and is used in this study for detecting proton flux through F0F1-ATPase within chromatophores driven by ATP hydrolysis. F-DHPE is easily labeled to the outer surface of chromatophores. In the range of pH 7.0 to 9.0, fluorescence intensity is sensitive to pH changes. The sensitivity is especially great in the range of pH 8.2 to 9.0, so pH 8.6 was chosen as the appropriate experimental condition. It is shown that added ATP not only acts as a fluorescence quencher but also can be hydrolyzed by F0F1-ATPase to pump protons into chromatophores, resulting in fluorescence restoration. A stimulator (NaSO3) and various types of inhibitors (NaN3, 5'-adenylyl imidodiphosphate [AMP-PNP], and N,N'-dicyclohexylcarbodiimide [DCCD]) of F0F1 confirmed that fluorescence restoration is caused by ATP-driven proton flux. When loaded with one antibody (anti-beta antibody) or two antibodies (anti-beta antibody and sheep to rabbit second antibody), F0F1-ATPase exhibits lower proton pumping activities, as indicated by fluorescence restoration. The possible mechanism of the inhibition of antibodies on proton pumping activity is discussed. PMID- 16043114 TI - [Laparoscopic Janeway gastrostomy: an advantageous solution for self-sufficient enteral feeding]. AB - INTRODUCTION: Laparoscopic gastrostomy according to Janeway (LGJ) is an alternative to percutaneous gastrostomy techniques. METHODS: A series of 10 LGJ is reported. The laparoscopic technique involves an isoperistaltic tube of 6-7 cm of length and 10-12 mm of diameter is created by 2 applications of linear stapling and cutting device. The tube is led out, opened and fixed to the fascial and cutaneous planes and a Foley catheter is inserted. RESULTS: Mean operation time was 35 minutes. There was no complication. The LGJ was indicated in 9 patients with tumour of the pharynx and 1 patient with encephalopathy. CONCLUSION: The main drawback of the LGJ is the need of general anaesthesia. The main advantage is the creation by minimal invasive surgery of a permanent gastrostomy equipped with a removable catheter easily changeable by non specialized health professionals, and even by the patient himself. PMID- 16043115 TI - [Reducing the hospital stay after colorectal resection]. AB - INTRODUCTION: The postoperative hospital stay after colorectal resection is about 15 days in France, when some authors have published a postoperative stay of 2 to 5 days. The aim of this work was to obtain a postoperative hospital stay less than 7 days. PATIENTS AND METHODS: Sixty-one patients who underwent a colorectal resection performed by laparotomy were included in the study: 16 right hemicolectomies, 9 left hemicolectomies, 15 sigmoidectomies and 21 anterior resections were performed. The operation was performed through a midline incision extended over the umbilicus in 13 cases, limited below the umbilicus in 22 cases and elective in 26 cases (right transverse in 16 and left iliac fossa in 10 cases). The protocol comprised epidural analgesia or wound infusion with ropivacaine, restricted intravenous fluids, early oral feeding and active mobilisation. RESULTS: The median and mean times of discharge were 6 and 7.3 days respectively; 36 patients (59%) were discharged on postoperative days 3 to 6, 8 patients (13%) on days 7 and 17 (28%) after day 7. A nasogastric tube was necessary in 2 cases (3.3%). Ten (16%) postoperative complications and 3 (5%) readmissions occurred. There were no deaths. CONCLUSION: Although the postoperative stay cannot be reduced in all the cases, a median hospital stay inferior to which is currently observed can easily be obtained by applying some simple and inexpensive means. This is advantageous for the patient, whose recovery is faster, and contributes to reduce the cost, which is of crucial importance today. PMID- 16043116 TI - [Bladder herniation]. AB - Inguinal hernia of the bladder is uncommon. The diagnosis is frequently done during surgery. It can be evoked before surgery when the patient presents with irritative and obstructive lower urinary tract symptoms. Retrograde urethrocystography or computerized tomography confirm the diagnosis. We report a case of bilateral inguinal hernia involving the bladder at the right side in a 87 years old man who had a surgical history of transvesical adenomectomy of the prostate. The diagnosis was done on the retrograde urethrocystography. We performed bilateral herniorrhaphy with herniated bladder reduction. The postoperative history was uneventful. PMID- 16043117 TI - The size of the population of weakly coupled chlorophyll pigments involved in thylakoid photoinhibition determined by steady-state fluorescence spectroscopy. AB - On the basis of experiments with singlet quenchers and in agreement with previous data, it is suggested that a population of energetically weakly coupled chlorophylls may play a central role in photoinhibition in vivo and in vitro. In the present study, we have used steady state fluorescence techniques to gain direct evidence for these uncoupled chlorophylls. Due to the presence of their emission maxima, near 650 nm and more prominently in the 670--675 nm interval both chlorophylls b and a seem to be involved. A straightforward mathematical model is developed to describe the data which allows us to conclude that the uncoupled/weakly coupled population size is in the range of 1--3 molecules per photosystem. PMID- 16043118 TI - When the heat is on: romantic partner responses influence distress in socially anxious women. AB - This study provided a preliminary test of whether socially anxious women and their partners would show more negative social support behavior, especially among those with low relationship satisfaction, and whether this would increase the distress of the socially anxious women. Women with (n=22) and without (n=23) heightened social anxiety were observed interacting with their partners under a social-evaluative threat and support behaviors were coded. Unexpectedly, no differences were found between socially and non-socially anxious women and their partners. Although relationship satisfaction influenced this process, it was the more satisfied women who showed more negative behavior. Additionally, the more positive behaviors the partner exhibited, the greater was the distress reported by socially anxious women, particularly among women whose partners reported high relationship satisfaction. Implications for how these findings might expand theories on interpersonal processes in social anxiety are discussed. PMID- 16043119 TI - Dynamics of calcium fluxes in human platelets assessed in calcium-free medium. AB - Dynamics of changes in cytosolic calcium concentration resulting from facilitation of calcium leakage from the stores and (or) blocking the pathways of its reuptake back into the stores or extrusion out of the cell (or both) have been investigated experimentally. It has been found that: (a) no mechanisms other than the membrane leakage, PMCA or SERCA, are involved in the discharge of calcium stores and calcium extrusion or reuptake; (b) the discharge of calcium stores in the absence of both its extrusion and reuptake back into the stores depends only on membrane leakage, the asymptotic calcium concentration in cytosol depending only on the initial content of the stores and being independent of the leakage; (c) the dynamics of the activity of both PMCA and SERCA depend on the initial rate of calcium influx, the dynamics differing from each other at high initial rates of calcium influx; (d) whereas there is no observable background activity of PMCA, background activity of SERCA is observed. PMID- 16043120 TI - Binding of two zinc finger nuclease monomers to two specific sites is required for effective double-strand DNA cleavage. AB - Custom-designed zinc finger nucleases (ZFNs) are becoming powerful tools in gene targeting-the process of replacing a gene within a genome by homologous recombination. Here, we have studied the DNA cleavage by one such ZFN, DeltaQNK FN, in order to gain insight into how ZFNs cleave DNA and how two inverted sites promote double-strand cleavage. DNA cleavage by DeltaQNK-FN is greatly facilitated when two DeltaQNK-binding sites are close together in an inverted orientation. Substrate cleavage was not first order with respect to the concentration of DeltaQNK-FN, indicating that double-strand cleavage requires dimerization of the FokI cleavage domain. Rates of DNA cleavage decrease as the substrate concentrations increase, suggesting that the DeltaQNK-FN molecules are effectively "trapped" in a 1:1 complex on DNA when the DNA is in excess. The physical association of two ZFN monomers on DNA was monitored by using the biotin pull-down assay, which showed that the formation of DeltaQNK-FN active complex required both binding of the two DeltaQNK-FN molecules to specific DNA sites and divalent metal ions. PMID- 16043121 TI - CXCR3 polymorphisms associated with risk of asthma. AB - The chemokine (C-X-C motif) receptor 3 (CXCR3) gene, on chromosome Xq13, is known to have critical roles in inflammatory and immune responses. In an effort to discover polymorphisms have been implicated in asthma, we investigated the genetic polymorphisms in CXCR3 to evaluate it as a potential candidate gene for a host genetic study of asthma. Statistical analysis revealed that one SNP in intron 1, c.12+234G > A, showed significant association with the risk of asthma development (P = 0.007, OR = 0.81). By subgroup analyses stratified by gender and atopic status, the genetic effect of c.12+234G > A on asthma was more apparent among male atopic subjects (P = 0.0009, OR = 0.61). Our findings suggest that polymorphisms in CXCR3 might be one of the genetic factors for the risk of asthma development, especially in male atopic subjects. CXCR3 variation/haplotype information identified in this study will provide valuable information and insight into strategies for the control of asthma and its subgroup, atopy. PMID- 16043122 TI - CREB expression in cardiac fibroblasts and CREM expression in ventricular myocytes. AB - Activation of gene expression by the cAMP-dependent signaling pathway is regulated by members of the cAMP response element binding protein (CREB) family consisting of CREB, CREM, and ATF-1. It is decisively for the understanding of the heart function as to which type of heart cells expresses CREB and/or CREM. Ventricular myocytes and fibroblasts of young (3 months) and old (24 months) rat hearts were separately investigated to analyse CREB, CREM, and phospho-CREB. Western blot showed CREB expression exclusively in fibroblasts but CREM was predominantly detected in ventricular myocytes. CREB-positive nuclei in heart sections were only revealed in fibroblasts. CREB was activated by forskolin (10 microM), PMA (500 nM), and cyclical mechanical strain (1 Hz, 5% elongation) in fibroblasts. The number of CREB-positive myocytes in old rats was larger than in young rats. But CREB could not be activated by forskolin (10 microM) in all myocytes. Our results suggest that the expression of CREB depends on the cell type and the age of the animal. We discuss that modulation of gene expression as it occurs with a age could be affected by the change within the CREB family members. PMID- 16043123 TI - Highly specific expression of luciferase gene in lungs of naive nude mice directed by prostate-specific antigen promoter. AB - PSA promoter has been demonstrated the utility for tissue-specific toxic gene therapy in prostate cancer models. Characterization of foreign gene overexpression in normal animals elicited by PSA promoter should help evaluate therapy safety. Here we constructed an adenovirus vector (AdPSA-Luc), containing firefly luciferase gene under the control of the 5837 bp long prostate-specific antigen promoter. A charge coupled device video camera was used to non-invasively image expression of firefly luciferase in nude mice on days 3, 7, 11 after injection of 2 x 10(9)PFU of AdPSA-Luc virus via tail vein. The result showed highly specific expression of the luciferase gene in lungs of mice from day 7. The finding indicates the potential limitations of the suicide gene therapy of prostate cancer based on selectivity of PSA promoter. By contrary, it has encouraging implications for further development of vectors via PSA promoter to enable gene therapy for pulmonary diseases. PMID- 16043124 TI - Transport mechanisms for adenosine and uridine in primary-cultured rat cortical neurons and astrocytes. AB - Endogenous adenosine and uridine are important modulators of neural survival and activity. In the present study, we examined transport mechanisms of adenosine and uridine in primary-cultured rat cortical neurons, and compared the results for neurons with those for astrocytes. Reverse transcription-polymerase chain reaction identified the mRNAs for ENT1, ENT2, and CNT2, but not CNT1 and CNT3, in neurons and astrocytes. [3H]Adenosine and [3H]uridine were time-, temperature-, and concentration-dependently taken up into neurons and astrocytes. In kinetic analyses, the uptake of both substrates by neurons and astrocytes consisted of two and one, respectively, saturable transport components. The uptake clearance for both substrates by neurons was greater than that by astrocytes. The relative contribution of the high-affinity major component of both substrates to total uptake was estimated to be approximately 80% in neurons. The uptake of [3H]adenosine and [3H]uridine by both neurons and astrocytes was almost entirely Na+-independent, and sensitive to micro, but not nano, molar concentrations of nitrobenzylmercaptopurine riboside, which are transport characteristics of ENT2. Therefore, it was indicated that adenosine and uridine are more efficiently taken up into neurons than into astrocytes, and ENT2 may predominantly contribute to the transport of the nucleosides as a high-affinity transport system in neurons, as in the case of astrocytes. PMID- 16043125 TI - Deregulated GSK3beta activity in colorectal cancer: its association with tumor cell survival and proliferation. AB - Glycogen synthase kinase 3beta (GSK3beta) reportedly has opposing roles, repressing Wnt/beta-catenin signaling on the one hand but maintaining cell survival and proliferation through the NF-kappaB pathway on the other. The present investigation was undertaken to clarify the roles of GSK3beta in human cancer. In colon cancer cell lines and colorectal cancer patients, levels of GSK3beta expression and amounts of its active form were higher in tumor cells than in their normal counterparts; these findings were independent of nuclear accumulation of beta-catenin oncoprotein in the tumor cells. Inhibition of GSK3beta activity by phosphorylation was defective in colorectal cancers but preserved in non-neoplastic cells and tissues. Strikingly, inhibition of GSK3beta activity by chemical inhibitors and its expression by RNA interference targeting GSK3beta induced apoptosis and attenuated proliferation of colon cancer cells in vitro. Our findings demonstrate an unrecognized role of GSK3beta in tumor cell survival and proliferation other than its predicted role as a tumor suppressor, and warrant proposing this kinase as a potential therapeutic target in colorectal cancer. PMID- 16043126 TI - A novel Gcn5p inhibitor represses cell growth, gene transcription and histone acetylation in budding yeast. AB - Histone acetyltransferases are key chromatin regulators responsible for transcriptional activation and cell cycle progression. We propose a simple yeast based assay to determine the specificity and targets of novel Gcn5p inhibitors. Here, we report the finding of a novel, small molecule, MC1626, which is able to inhibit yeast cell growth, Gcn5p-dependent gene transcription and acetylation of the histone H3 N-terminal tail in vivo. Because HATs misregulation is invariably associated with human diseases, the identification of MC1626 as a novel cell permeable Gcn5p inhibitor suggests that it may be a very useful starting tool for the further development of new molecules to be applied to expression profiling of genes regulated by histone H3 acetylation. In addition, our results demonstrate that MC1626 is a Gcn5p-dependent yeast growth inhibitor. PMID- 16043127 TI - Allosteric modulation of ligand-gated ion channels. AB - Ligand-gated ion channels (LGICs) are cell surface proteins that play an important role in fast synaptic transmission and in the modulation of cellular activity. Due to their intrinsic properties, LGICs respond to neurotransmitters and other effectors (e.g. pH) and transduce the binding of a ligand into an electrical current on a microsecond timescale. Following activation, LGICs open allowing an ion flux across the cell membrane. Depending upon the charge and concentration of ions, the flux can cause a depolarization or hyperpolarization, thus modulating excitability of the cell. While our understanding of LGICs has significantly progressed during the past decade, many properties of these proteins are still poorly understood, in particular their modulation by allosteric effectors. LGICs are often thought as a simple on-off switches. However, a closer look at these receptors reveals a complex behavior and a wide repertoire of subtle modulation by intrinsic and extrinsic factors. From a physiological point of view, this modulation can be seen as an additional level of complexity in the cell signaling process. Here we review the allosteric modulation of LGICs in light of the latest findings and discuss the suitability of this approach to the design of new therapeutic molecules. PMID- 16043128 TI - The neural correlates of anhedonia in major depressive disorder. AB - BACKGROUND: Anhedonia is a relative lack of pleasure in response to formerly rewarding stimuli. It is an important diagnostic feature of major depressive disorder (MDD), and predicts antidepressant efficacy. Understanding its neurobiological basis may help to target new treatments and predict treatment outcomes. Using a novel paradigm, we aimed to explore the correlations between anhedonia severity and magnitude of neural responses to happy and sad stimuli in regions previously implicated in studies of human reward processing and depressive anhedonia. METHODS: Neural responses to happy and sad emotional stimuli (autobiographical prompts and mood congruent facial expressions) were measured using blood oxygen level dependent (BOLD) functional magnetic resonance imaging in twelve MDD individuals with varying degrees of anhedonia. RESULTS: In response to happy stimuli, anhedonia, but not depression severity per se, was positively and negatively correlated with ventromedial prefrontal cortex (VMPFC) and amygdala/ventral striatal activity, respectively. State anxiety independently contributed to a VMPFC-subcortical dissociation of response to happy (but not sad) stimuli, which was similar, but different, to anhedonia. CONCLUSIONS: These findings suggest that anhedonia and state anxiety are associated with dysfunction within neural systems underlying the response to, and assessment of, the rewarding potential of emotive stimuli in MDD, and highlight the importance of employing a symptom-dimension-based approach in the examination of the neurobiology of depression. PMID- 16043129 TI - Studies of humoral immunity to preprohypocretin in human leukocyte antigen DQB1*0602-positive narcoleptic subjects with cataplexy. AB - BACKGROUND: Canine models for narcolepsy have mutations of the hypocretin receptor 2 gene, and preprohypocretin knockout murine lines exhibit narcoleptic like behaviors. Human narcolepsy with cataplexy is associated with human leukocyte antigen DQB1*0602 and reduced hypocretin levels in cerebrospinal fluid, suggesting an autoimmune diathesis. We tested the hypothesis that DQB1*0602 positive narcoleptic subjects with cataplexy have immunoglobulin (Ig)G reactive to human preprohypocretin and its cleavage products. METHODS: Serum samples of 41 DQB1*0602-positive narcoleptic subjects with cataplexy and 55 control subjects were studied, as were 19 narcoleptic and 13 control samples of cerebrospinal fluid. We tested for IgG reactive to preprohypocretin and its major cleavage products (including hypocretin 1 and 2), using immunoprecipitation assays (IP), immunofluorescence microscopy (IF) of Chinese hamster ovarian cells expressing preprohypocretin, and Western blots. RESULTS: There was no evidence for IgG reactive to preprohypocretin or its cleavage products in CSF of subjects with narcolepsy as measured by IPs, Western blots, and IF. Although the IP with CSF and the C-terminal peptide showed significant differences by two methods of comparison, the control subjects had higher counts per minute than narcoleptic subjects, which was opposite to our hypothesis. CONCLUSIONS: The hypothesis that DQB1*0602-positive narcoleptic subjects with cataplexy have IgG reactive to preprohypocretin or its cleavage products was not supported. PMID- 16043130 TI - The Val66Met coding variant of the brain-derived neurotrophic factor (BDNF) gene does not contribute toward variation in the personality trait neuroticism. AB - BACKGROUND: The val66met variant located within the brain-derived neurotrophic factor gene (BDNF) has previously been associated with human neuroticism, a dimension of personality strongly predictive of depressive illness. METHODS: Here we report an attempt to replicate this association using three populations of extreme neuroticism scorers derived from two large English cohorts (n = 88,142 and n = 20,921). On the basis of the current literature, which indicates that an effect of BDNF may only become apparent in those individuals exposed to stress, a gene-environment interaction was also sought. RESULTS: No statistically significant effects were identified, although simulations indicated that the samples held sufficient power to detect a main effect accounting for just .75% of variation and an interaction accounting for 4% of variation. CONCLUSIONS: These data do not support the hypothesis that the val66met BDNF polymorphism contributes toward variation in the human personality trait neuroticism, at least as indexed by the Eysenck Personality Questionnaire. PMID- 16043131 TI - Polymorphisms in the promoter region of the alpha1A-adrenoceptor gene are associated with schizophrenia/schizoaffective disorder in a Spanish isolate population. AB - BACKGROUND: Animal models have implicated the alpha(1)-adrenergic subtypes in cognitive functions relevant to schizophrenia, but no consensus exists with regard to the status of noradrenergic receptor populations in psychiatric patients. We focused on one alpha(1)-adrenergic subtype, the alpha(1A)-adrenergic receptor, and proposed that genetic variants within the regulatory region of this gene (ADRA1A) alter the expression of this receptor, influencing susceptibility toward schizophrenia. METHODS: This study examined this proposal by testing the hypothesis that single nucleotide polymorphisms (SNPs) in the promoter region of the alpha(1A)-adrenergic gene were associated with schizophrenia by performing case-control association analysis on SNPs found in a 5' upstream region, which included the putative promoter region and 5' untranslated region. Our sample consisted of 103 schizophrenia and 14 schizoaffective disorder patients and 176 control subjects. All recruits were from a Spanish population isolate of Basque origin that is characterized by low heterogeneity, which was selected with the intent that it might facilitate the identification of disease-related polymorphisms. RESULTS: A total of eight SNPs (-9625 G/A, -7255 A/G, -6274 C/T, 4884 A/G, -4155 C/G, -2760 A/C, -1873 G/A, and -563 C/T) were confirmed at a rare allele frequency of >5%. Association with schizophrenia and schizoaffective disorder was found for the -563 C/T SNP (p = .0005 for allele and p = .007 for genotype, Bonferroni corrected) and -9625 G/A SNP (p = .02 for allele and p = .03 for genotype, Bonferroni corrected). Significant differences in the 54 haplotypes formed by these eight SNPs were also found between patients and control subjects (p = .008, Bonferroni corrected). CONCLUSIONS: Because of the strength of these results and the location of these SNPs in the regulatory region of this gene, functional studies investigating the possible influence of these SNPs on receptor expression levels in schizophrenia are warranted. PMID- 16043132 TI - Brain regions associated with the expression and contextual regulation of anxiety in primates. AB - BACKGROUND: A key to successful adaptation is the ability to regulate emotional responses in relation to changing environmental demands or contexts. METHODS: High-resolution PET 18fluoro-deoxyglucose (FDG) scanning in rhesus monkeys was performed during two contexts (alone, and human intruder with no eye contact) during which the duration of anxiety related freezing behavior was assessed. Correlations between individual differences in freezing duration and brain activity were performed for each of the two conditions, as well as for the contextual regulation between the two conditions. RESULTS: In both conditions, activity in the basal forebrain, including the bed nucleus of the stria terminalis and the nucleus accumbens were correlated with individual differences in freezing duration. In contrast, individual differences in the ability to regulate freezing behavior between contexts were correlated with activity in the dorsal anterior cingulate cortex, the thalamus and the dorsal raphe nucleus. CONCLUSIONS: These findings demonstrate differences in the neural circuitry mediating the expression compared to the contextual regulation of freezing behavior. These findings are relevant since altered regulatory processes may underlie anxiety disorders. PMID- 16043133 TI - Catechol-O-methyltransferase (COMT) genotypes and working memory: associations with differing cognitive operations. AB - BACKGROUND: Catechol-O-methyltransferase (COMT) is a strong candidate gene for schizophrenia and cognitive functions disrupted in this disorder. This report examines the relation of COMT genotypes to performance on a battery of working memory tests differing in the cognitive operations to be performed on the material. METHODS: A large sample of 402 healthy adults were tested on four working memory tests: Spatial Delayed Response (SDR), Word Serial Position Test (WSPT), N-back, and Letter-Number Sequencing. A subsample (n = 246) was tested on the Wisconsin Card Sorting Test (WCST). A saliva swab was used to obtain DNA from all participants. RESULTS: Letter-Number Sequencing, which requires both storage and manipulation of information, was the only working memory test that showed expected differences among COMT genotypes, with the met/met group showing the best performance and the val/val group the poorest performance. As in previous studies, the met/met group also performed better than the val/val group on the WCST. CONCLUSIONS: COMT genotypes were not associated with performance on tests measuring simple storage, maintenance of temporal order or updating of information in working memory. Genotype differences in Letter-Number Sequencing and WCST suggest that higher-order components of processing (e.g., mental manipulation) are more closely related to this gene. PMID- 16043134 TI - Diffusion tensor imaging in schizophrenia. AB - BACKGROUND: Diffusion tensor imaging (DTI) is a relatively new neuroimaging technique that can be used to examine the microstructure of white matter in vivo. A systematic review of DTI studies in schizophrenia was undertaken to test the hypothesis that DTI can detect white matter differences between schizophrenia patients and normal control subjects. METHODS: EMBASE, PubMed, Medline, and PsychInfo were searched online and key journals were searched manually for studies comparing anisotropy (a measure of white matter integrity) between patients and control subjects. Nineteen articles were systematically reviewed. RESULTS: Though 16 studies found differences, methodological and data differences prevented a meta-analysis. Fourteen studies found reduced anisotropy in patients; two studies found only a loss of normal asymmetry. The region of investigation varied across studies, however, and when the same region (for example, the cingulum) was examined in different studies, as many failed to find a difference as found one. These inconsistencies may be the result of small sample sizes and differences in methodology. CONCLUSIONS: Diffusion tensor imaging has yet to provide consistent findings of white matter abnormalities in schizophrenia. Its potential as a means of examining anatomical connectivity may be realized with the study of larger, more homogenous groups of subjects and with ongoing improvements in image analysis. PMID- 16043135 TI - Evidence for deficit in tasks of ventral, but not dorsal, prefrontal executive function as an endophenotypic marker for bipolar disorder. AB - BACKGROUND: Trait functional abnormalities in BD patients have only been reported in the ventral prefrontal cortex (VPFC). We examined whether deficits in VPFC related inhibitory processes, but not dorsal prefrontal (DPFC) based executive functions, represent an endophenotypic marker for bipolar disorder I (BDI). METHODS: We used the Wisconsin Card Sorting Test (WCST), commonly associated with DPFC function, and the Hayling Sentence Completion Task (HSCT) which engages the VPFC. Performance on these tests of 43 healthy participants was compared to that of 10 remitted BDI patients and 15 of their unaffected offspring. RESULTS: Compared to healthy participants, patients and their offspring made more errors in the HSCT but offspring achieved more categories and made fewer perseverative errors in the WCST. CONCLUSIONS: Impaired response inhibition, predominantly a VPFC related function, may reflect familial predisposition to BDI while deficits in rule attainment, a DPFC based function, may be associated only with the clinical phenotype. PMID- 16043136 TI - The beta-1 adrenergic antagonist, betaxolol, improves working memory performance in rats and monkeys. AB - BACKGROUND: Previous studies have indicated that beta adrenergic receptor stimulation has no effect on the cognitive functioning of the prefrontal cortex (PFC). Blockade of beta-1 and beta-2 receptors in the PFC with the mixed beta 1/beta-2 antagonist, propanolol, had no effect on spatial working memory performance. However, more selective blockade of beta-1 or beta-2 receptors might show efficacy if the two receptors have opposite effects on PFC function. The current study examined the effects of the selective beta-1 antagonist, betaxolol, on working memory in rats and monkeys. METHODS: In rats, betaxolol (.0011-1.11 microg/.5 microL) was infused into the PFC 5 min before delayed alternation testing. Monkeys were systemically injected with betaxolol (.0000011-.11 mg/kg) 2 hours before delayed response testing. RESULTS: Betaxolol produced a dose-related improvement in working memory performance following either direct PFC infusion in rats, or systemic administration in monkeys. However, some aged monkeys developed serious pancreatic problems over the course of this study. CONCLUSIONS: These findings suggest that endogenous activation of the beta-1 adrenergic receptor impairs PFC cognitive function. These results may have therapeutic relevance to post-traumatic stress disorder or other disorders with excessive noradrenergic activity and PFC dysfunction. Pancreatic side effects in aged subjects taking betaxolol warrants further investigation. PMID- 16043137 TI - Normal prefrontal gamma-aminobutyric acid levels in remitted depressed subjects determined by proton magnetic resonance spectroscopy. AB - BACKGROUND: There is growing evidence that the brain gamma-aminobutyric acid (GABA) system is involved in depression. Lowered plasma GABA levels were identified as a traitlike abnormality found in patients with remitted unipolar depression and in healthy first-degree relatives of patients with unipolar depression. Major depressive disorder has been associated with neuroimaging and neuropathological abnormalities in the prefrontal cortex by various types of evidence. As a result, the current study investigates whether GABA levels in the prefrontal cortex differ between unmedicated subjects with remitted major depressive disorder (rMDD) and healthy control subjects. METHODS: Sixteen rMDD subjects and 15 healthy control subjects underwent magnetic resonance spectroscopy. We used a 3 Tesla GE whole body scanner with a homogeneous resonator coil providing a homogenous radiofrequency field and capability of obtaining measurement from the prefrontal cortex. Gamma-aminobutyric acid levels were measured in the ventromedial prefrontal cortex and dorsolateral/anterior medial prefrontal cortex. RESULTS: There was no difference in GABA concentrations between rMDD subjects and healthy control subjects in the ventromedial prefrontal cortex and dorsolateral/anterior medial prefrontal cortex. Secondary analyses provided preliminary evidence for a negative relationship between the glutamate/glutamine (Glx)/GABA ratio and age of onset of major depression in the ventromedial prefrontal cortex. CONCLUSIONS: This result suggests that GABA levels in the prefrontal cortex, if found to be reduced in symptomatic depression, do not represent a persistent characteristic of major depression. Further research is needed to determine brain GABA levels in different brain regions, in different stages of depressive illness, and in different depressive subtypes. PMID- 16043138 TI - Electroconvulsive seizures induce angiogenesis in adult rat hippocampus. AB - BACKGROUND: Electroconvulsive seizure (ECS)-treatment, a model for electroconvulsive therapy (ECT) has been shown to induce proliferation of endothelial cells in the dentate gyrus (DG) of adult rats. Here we quantified the net angiogenic response after chronic ECS-treatment in the molecular layer (ML) of the dentate gyrus. Patients undergoing ECT are routinely oxygenated to prevent hypoxia, a known inducer of angiogenesis. Therefore we also examined the effect of oxygenation on ECS-induced proliferation of endothelial cells. METHODS: Total endothelial cell numbers and vessel length were estimated utilizing design based stereological analysis methods. Endothelial cell proliferation in the DG after ECS with or without oxygenation was assessed using bromodeoxyuridine. RESULTS: The total number of endothelial cells and total vessel length was increased. Oxygenation did not abolish the ECS-induced proliferation of endothelial cells in the DG. CONCLUSIONS: ECS-treatment induces a dramatic increase in endothelial cell proliferation leading to a 30% increase in the total number of endothelial cells. The increase in cell number resulted in a 16% increase in vessel length. These findings raise the possibility that similar vascular growth is induced by clinically administered ECT. PMID- 16043141 TI - Genetic susceptibility to keloid disease: mutation screening of the TGFbeta3 gene. AB - Keloid disease (KD) is a fibroproliferative dermal tumour of unknown aetiology. The increased familial clustering in KD, its increased prevalence in certain races and its presence in identical twins suggest a strong genetic predisposition to keloid formation. Transforming growth factor beta isoforms (TGFbeta) play a central role in wound healing and fibrosis and have been implicated in KD pathogenesis. Recent data has suggested that TGFbeta(3) has an important role in scar formation. There is little known about the genetic variation present within the TGFbeta(3) gene, which contains seven exons and six introns spanning 43,000 base pairs of the human genome. Exons one to seven and the promoter region (1000 bp upstream from exon 1 in the 5'-flanking regions) were screened in 95 Caucasian KD cases and 95 Caucasian controls for the presence of novel mutations using a high throughput DHPLC mutation detection technology. There were no mutations identified in any of the exonic regions, however, multiple nondisease associated mutations were found in the promoter region of the TGFbeta(3) gene. These data demonstrate that there is no association between the exonic and promoter regions of TGFbeta(3) gene and keloid scarring in our cohort of Caucasian patients. PMID- 16043139 TI - Disparity in the temporal appearance of methamphetamine-induced apoptosis and depletion of dopamine terminal markers in the striatum of mice. AB - Methamphetamine (METH) causes damage in the striatum at pre- and post-synaptic sites. Exposure to METH induces long-term depletions of dopamine (DA) terminal markers such as tyrosine hydroxylase (TH) and DA transporters (DAT). METH also induces neuronal apoptosis in some striatal neurons. The purpose of this study is to demonstrate which occurs first, apoptosis of some striatal neurons or DA terminal toxicity in mice. This is important because the death of striatal neurons leaves the terminals in a state of deafferentation. A bolus injection (i.p.) of METH (30 mg/kg) induces apoptosis (TUNEL staining) in approximately 25% of neurons in the striatum at 24 h after METH. However, in contrast to apoptosis, depletion of TH (Western blotting) begins to appear at 24 h after METH in dorsal striatum while the ventral striatum is unaffected. The peak of TH depletion (approximately 80% decrease relative to control) occurs at 48 h after METH. Autoradiographic analysis of DAT sites showed that depletion begins to appear 24 h after METH and peaks at 2 days (approximately 60% depletion relative to control). Histological analysis of the induction of glial fibrillary acidic protein (GFAP) by METH in striatal astrocytes revealed an increase at 48 h after METH that peaked at 3 days. These data demonstrate that striatal apoptosis precedes the depletion (toxicity) of DA terminal markers in the striatum of mice, suggesting that the ensuing state of deafferentation of the DA terminals may contribute to their degeneration. PMID- 16043142 TI - Remodelling the upper eyelid in the management of orbitopalpebral neurofibromatosis. AB - Neurofibromatosis of the orbitopalpebral complex is a debilitating disease. The development of safe craniofacial surgical technique has greatly improved outcome in the surgical palliation of this progressive condition, and various centres have reported their experience. Extensive upper eyelid neurofibroma creates a functional and aesthetic problem, which is inadequately addressed in the literature. The experience of the senior author (DM) representing the treatment of 27 such patients is currently reported. The stigmata of eyelid neurofibromatosis were scored retrospectively by a novel scale applied to standardised photographs at patient presentation. A numeric severity score was given to each of: upper eyelid ptosis, canthal malposition, and oculo-palpebral diastasis. Disease progression and post-operative result were similarly scored at various stages follow-up photography. All patients underwent a full thickness, transverse or transverse-oblique resection of upper eyelid disease, with immediate levator reconstruction and canthopexies as necessary. A mean improvement of 3.33 points was achieved in the majority of patients (median one operation per patient; mean 1.5, range 1-3). Secondary eyelid procedures (n=9 pts) represented tumour debulking, canthopexy, or procedures to deepen the fornix in patients with prostheses. This series demonstrates that a tailored, often radical palpebral remodelling in orbitopalpebral neurofibromatosis may provide functional and aesthetic benefit. PMID- 16043143 TI - Scalp reconstruction with free flaps using the external carotid artery and internal jugular vein as the recipient vessels without the need for interpositional vein grafts or arterio-venous loops. AB - The technique of lengthening of the pedicles of the rectus abdominus and latissimus dorsi free flaps are described in five patients. Lengthening of these pedicles allowed scalp reconstruction using the external carotid artery and internal jugular vein as the recipient vessels without the need for interpositional vein grafts or arterio-venous loops. Post-operative recovery was uneventful in all cases. PMID- 16043144 TI - Nipple migration in a pig using the technique of serial excision. AB - Moving a misplaced nipple-areola complex (NAC) without causing additional scarring has proved to be difficult, especially if it is to be moved medially, laterally or inferiorly. This study investigated the possibility of migrating the NAC without leaving additional scarring by using the technique of serial excision. Two adult female pigs were used. Three pairs of pseudo-areolas of similar size were tattooed around a teat in each pig. Crescentic serial excisions of skin were undertaken at 2 weekly intervals on the upper and middle pairs of pseudo-areolas in each pig and the lower pairs were left as controls. The crescentic excision on the upper set of nipples consisted of skin only. The crescentic excision on the middle set of nipples consisted of 75% skin and 25% tattooed pseudo-areola. Results showed that nipple migration by serial excision is possible. However, each set of nipple pseudo-areola complex (NPAC) behaved differently. When skin only was excised, the NPAC's became distorted and enlarged but when skin and areola was excised, the NPAC maintained its dimensions. Taking our findings into account, an approach to moving a misplaced NAC using this technique in the clinical situation is described. Its use to move a misplaced NAC in two women is demonstrated. PMID- 16043146 TI - Potential blindness: an unusual complication of lateral canthopexy. AB - Lateral canthopexy is a commonly performed procedure in craniofacial and cosmetic surgery. In craniofacial surgery, lateral canthal fixation is performed in conjunction with a wide range of transcranial or subcranial procedures during the process of soft tissue re-suspension. A number of fixation materials have gained popular use. A case of craniofrontonasal dysplasia is reported in which a wire loop canthopexy fixation has become disengaged 3 months after a history of trauma and rotated to present a sharp surface against the sclera. Urgent surgical exploration prevented the apparently imminent complication of globe penetration with associated threat to vision. PMID- 16043147 TI - Horse shoe flap vaginoplasty--a new technique of vaginal reconstruction with labia minora flaps for primary vaginal agenesis. AB - Reconstruction of the vagina has always been a formidable surgical challenge. A variety of procedures with varying degrees of success have been described, but none has proved to be the ideal method. We have done 15 vaginal reconstructions using labia minora flaps. In the first eight patients, we used two labia minora flaps and sutured them together to create a neovagina. In the last eight patients this new technique of vaginoplasty using horse shoe labia minora flap was used. The neovagina is dilated gradually for a period of 3 months and at the end of 3 months the patient has an adequate sized vagina. All were done for vaginal agenesis. Horse shoe flap vaginoplasty in Mayer, Rokitansky-Kuster-Hauser syndrome is a new technique. The entire hairless skin within the labia majora is used to create a neovagina. Both the labia minora along with the prepucial skin of the clitoris is elevated as a single flap thereby increasing the dimension of the Neovagina and also augmenting the blood supply of the flap, making it more reliable and finally the design simplifies the technique of suturing the neovaginal tube. PMID- 16043148 TI - The use of pimonidazole to characterise hypoxia in the internal environment of an in vivo tissue engineering chamber. AB - The distribution of hypoxic cells in an in vivo tissue engineering chamber was investigated up to 28 days post-implantation. METHODS: Arteriovenous loops were constructed and placed into bi-valved polycarbonate chambers containing 2 x 10(6) rat fibroblasts in basement membrane gel (BM gel). Chambers were inserted subcutaneously in the groin of male rats and harvested at 3 (n = 6), 7 (n = 6), 14 (n = 4) or 28 (n = 4) days. Ninety minutes before harvest, pimonidazole (60 mg/kg) was injected intraperitoneally. Chamber tissue was removed, immersion fixed, paraffin embedded, sectioned and stained immunohistochemically using hypoxyprobe-1 Mab that detects reduced pimonidazole adducts forming in cells, where pO2 < 10 mmHg. RESULTS: At 3 days a fibrin clot/BM gel framework filled the chamber. Seeded fibroblasts had largely died. The majority of 3 day chambers did not demonstrate tissue growth from the AV loop nor was pimonidazole binding present in these chambers. In one chamber in which tissue growth had occurred strong pimonidazole binding was evident within the new tissue. In four out of six 7 day chambers a broader proliferative zone existed extending up to 0.4 mm (approximately) from the AV loop endothelium which demonstrated intense pimonidazole binding. The two remaining 7 day chambers displayed even greater tissue growth (leading edge > 0.7 mm from the AV loop endothelium), but very weak or no pimonidazole binding. At 14 and 28 days the fibrin/BM gel matrix was replaced by mature vascularised connective tissue that did not bind pimonidazole. CONCLUSION: Employing a tissue engineering chamber, new tissue growth extending up to 0.4 mm from the AV loop endothelium (chambers < or = 7 days) demonstrated intense pimonidazole binding and, therefore, hypoxia. Tissue growth greater than 0.5 mm from the AV loop endothelium (7-28 days chambers) did not exhibit pimonidazole binding due to a significant increase in the number of new blood vessels and was, therefore, adequately oxygenated. PMID- 16043150 TI - Planning and use of therapeutic mammoplasty--Nottingham approach. AB - Therapeutic mammaplasty, the use of reduction mammaplasty and radiotherapy to surgically treat breast cancer, is an established technique for selected breast cancers and can extend the role of breast conserving surgery. Most frequently described is the use of a wise pattern reduction for tumours that lie within the expected mammaplasty excision. However, mammaplasty techniques can be safely adapted to treat patients with cancers in all areas of the breast. An approach to selection and planning surgery is presented which has evolved from the experience of other units and our first 50 clinical cases over a 3-year period. The outcomes of these 50 cases are found in the accompanying article. Technique will vary depending upon the tumour position. Breast cancers may lie within the normal excision site of a recognised mammaplasty method (scenario A) or outside of the expected excision sites (scenario B). In scenario A, a range of recognised techniques can be performed without adaptation to widely excise the tumour and re shape the breasts. In scenario B the techniques need to be adapted. Three decisions are needed for planning in scenario B; the skin incision, the nipple aereola complex (NAC) pedicle orientation and finally the method of filling the cancer defect. The latter can be achieved by either extending the nipple pedicle or by creating a secondary pedicle within the breast dissection. Either method will move tissue that is normally excised into the cancer defect. For central tumours an inferior pedicle is usually used to both fill the defect and re-create the nipple. PMID- 16043151 TI - The combined free partial vastus lateralis with anterolateral thigh perforator flap reconstruction of extensive composite defects. AB - Myocutaneous (MC) free flaps are useful for many reconstructive indications. Perforator flaps have become standard of care. The anterolateral thigh flap (ALT) donor site is popular. With the ALT flap varying sizes of vastus lateralis (VL) muscle can be harvested as a MC flap. The skin islands of these flaps have a great range of freedom when dissected on their perforator. It was hypothesised that the VL-ALT perforator flap would offer adequate tissue volume combining maximal freedom in planning with minimal donor site morbidity. From November 2001 to February 2003 a free partial VL with ALT perforator flap was used in 11 patients to reconstruct large defects. Indications for adding a muscular component were exposed bone, skull base, (artificial) dura, or osteosynthesis material, open sinuses, and lack of muscular bulk. Flaps were planned as standard ALT flaps, after which three types of dissection were performed: I. true MC flap; II. muscle flap with a skin island on one perforator, which could be rotated up to 180 degrees ; III. chimera skin perforator flap with muscle being harvested on a separate branch from the source vessel or on a side branch of the skin perforator. Mean skin size of the MC-ALT flaps was 131 cm2. Mean muscle part size of the MC-ALT flaps was 268 cm3. Muscular parts were custom designed for all defects. No total or partial flap failures were seen. Colour mismatch was seen in 6 of 8 patients, when skin was used in the facial area in this all white population. Excessive flap bulk was found in 8 of 11 patients at 6 weeks, however, only in 2 of 11 patients after 6 months. Patients were satisfied with the functional result (8 of 11 patients) as well as the cosmetic result of their reconstruction (7 of 11 patients). All less satisfied patients had received their flap for external facial skin reconstruction. Donor site morbidity was minimal. The combined free partial VL with ALT perforator flap proved valuable as a (chimera type) MC flap with maximal freedom of planning to meet specific reconstructive demands and minimal donor site morbidity. PMID- 16043152 TI - A vascularised bone graft from the medial femoral condyle for recurrent failed arthrodesis of the distal interphalangeal joint. AB - A vascularised bone graft from the medial femoral condyle was used to correct a recurrent failed arthrodesis of the index finger distal interphalangeal joint. The flap was based upon the articular branch of the descending genicular artery. Union was confirmed 3 months after surgery. PMID- 16043153 TI - Therapeutic mammaplasty--analysis of 50 consecutive cases. AB - This is a descriptive analysis of 50 consecutive cases of therapeutic mammaplasty, the use of reduction mammaplasty and radiotherapy to treat breast tumours. Breast cancers may lie within the normal excision site of a recognised mammaplasty method (scenario A) or outside of the expected excision sites (scenario B), then requiring a modified mammaplasty. A third group are central tumours requiring removal of the nipple. This series includes 13 scenario A, 27 scenario B and 10 central tumours. Wise pattern skin incision was used in 29 (58%) and vertical in 14 (28%). Seven different pedicles were employed but inferior (11), superior (9) and superio-medial (19) comprised the majority (78%). In the 27 scenario B cases an extension of the nipple aereolar pedicle was used to fill the tumour defect in 19 (70%) and a secondary pedicle in 8 (30%). There were no incomplete excisions of invasive tumour but incomplete excision of DCIS requiring mastectomy occurred in 4 (8%) patients. There are no deaths or recurrences to date (mean follow-up 13 months, range 3-32). Complications have occurred in 8 (16%) with a return to theatre for one complication (2%) and the four patients (8%) requiring mastectomy. No patient has had a delay in adjuvant treatment. Cosmetic outcome was deemed good/excellent in 63%, satisfactory in 33%, poor in 4%. Follow-up is restricted to 3-32 months. PMID- 16043154 TI - Giant basal cell carcinoma of the thoracic wall: a case report and review of the literature. AB - Giant basal cell carcinoma is a rare skin tumour with aggressive biological behaviour, and deep invasion and metastasis have been reported. The authors describe a giant basal cell carcinoma involving the anterior chest wall. The lesion infiltrated the mediastinum, occluding the left brachiocephalic vein. Vascular invasion caused venous occlusion affecting the left upper limb. Neither surgical treatment nor radiotherapy were practicable. PMID- 16043155 TI - Seven reconstructions of the orbital and periorbital region with a contralateral orbitonasolabial flap. AB - Blasius designed the first orbitonasolabial flap in 1842, which has since, been reshaped by many other surgeons. Three years ago, we were confronted with a difficult reconstructive case where ipsilateral local flaps were insufficient to cover a huge defect of the periorbital region. We performed 11 cadaver dissections to determine the type of vascularisation of the orbitonasolabial flap and to define its arc of rotation prior to contralateral use. This contralateral orbitonasolabial flap was subsequently applied to seven cases with satisfactory results. PMID- 16043156 TI - The role of the Lumina intense pulsed light system in the treatment of port wine stains--a case controlled study. AB - The pulsed dye laser has been the treatment of choice for port wine stains over the past 20 years. In the past 5 years there has been increasing discussion of the role of other light treatments, such as the intense pulsed light system in the treatment of port wine stains. These systems use high-energy lamps, which emit noncoherent broad-spectrum light. Cut off filters are applied depending on the treatment modality to limit the wavelengths emitted. We present the results of a 3-year prospective within patient controlled clinical trial using an intense pulsed light system called the Lumina, developed by Lynton Lasers of Cheshire, England. Our aims and objectives were to assess the effectiveness of the system in the treatment of port wine stains in a human model and to record the optimum treatment parameters and the incidence of side effects. Following ethical approval 12 subjects were enrolled into the trial. In order to meet the requirements of the local ethics committee these were all adults with port wine stains located in less visible areas of the body. The results showed that eight of the 12 subjects had some degree of fading of their port wine stain as measured on a percentage scoring system. Of the four who failed to show any response, all had pink port wine stains. It did seem the case that the darker the port wine stain, the better the fading seen. Furthermore, the more distal lesions tended to be less responsive than those situated closer to the head area. However, it is difficult to draw any definitive statistical conclusions due to the small number of patients in the trial. PMID- 16043157 TI - Bone marrow-impregnated collagen matrix for wound healing: experimental evaluation in a microcirculatory model of angiogenesis, and clinical experience. AB - OBJECTIVES: This study aims to investigate the effect of collagen matrix impregnated with bone marrow on wound healing angiogenesis in an effective microcirculatory model and to describe our clinical experience. METHODS: We used a skinfold chamber of original design which visualises microcirculation following wound creation on the dorsal skin of the mouse to establish an in vivo experimental model to estimate angiogenesis. Animals were divided into two groups: a bone marrow group (n = 6) in which bone marrow-impregnated collagen matrix was applied to the wound; and a control group (n = 7), in which collagen immersed in saline was applied, and functional capillary density was quantified during the repair process. RESULTS: The increase rate in functional capillary density during wound healing significantly increased in the bone marrow group on days 3, 5 and 7 after creation of the wound but no significant difference was detected on day 10. A patient with a chronic leg ulcer that had not responded to conventional therapy for 1 year was treated with autogenous bone marrow impregnated collagen matrix and successful wound closure was obtained. CONCLUSION: The present study suggested that collagen matrix impregnated with bone marrow significantly promoted the repair process, especially in the early stage. The features of the treatment, including the possible use of a patient's own cells, simple method, immediate application without any processing procedure and preservation of the inclusive potentiality of bone marrow suspension, offer significant advantages in terms of the anticipated routine clinical use. PMID- 16043158 TI - Synthesis of two bidesmosidic ursolic acid saponins bearing a 2,3-branched trisaccharide residue. AB - The focus of this work was on the synthesis of two bidesmosidic ursolic acid saponins bearing a 2,3-branched trisaccharide residue. Therefore, 3-O-{[beta-D glucopyranosyl-(1-->2)]-[alpha-L-arabinopyranosyl-(1-->3)]-alpha-L arabinopyranosyl}ursolic acid-28-O-[beta-D-glucopyranosyl] ester 1 was concisely synthesized by two strategies in 22% and 41% overall yield, respectively, and another congener 3-O-{[beta-D-xylopyranosyl-(1-->2)]-[beta-D-glucopyranosyl-(1- >3)]-alpha-L-arabinopyranosyl}ursolic acid-28-O-[beta-D-glucopyranosyl] ester 2 was also efficiently prepared in 81% overall yield. The (1)H NMR and (13)C NMR signals of saponin 2 are all consistent with those reported for the natural product. PMID- 16043159 TI - Crystal structure of methyl 3-amino-2,3-dideoxy-beta-D-arabino-hexopyranoside. Stabilization of the crystal lattice by a double network of N-H...O, O-H...N and O-H...O interactions. AB - The structure, conformation and configuration of methyl 3-amino-2,3-dideoxy-beta D-arabino-hexopyranoside were investigated by (1)H NMR, (13)C NMR and IR spectroscopy, as well as by optical rotation. The crystal structure was confirmed by single-crystal X-ray crystallographic analysis at 293 K and R = 0.0434 based on 910 independent reflections. The crystal belongs to the monoclinic system, space group of P2(1) with cell dimensions a = 6.050(1) Angstroms, b = 7.284(1) Angstroms, c = 10.289(2) Angstroms, beta = 104.69(3) degrees, D(c) = 1.341 Mg cm( 3) and V = 438.9(1) Angstroms(3) for Z = 2. Furthermore, the molecule has a typical (4)C(1) chair conformation. Hydrogen bonds between sugar molecules are responsible for stabilizing the crystal lattice. PMID- 16043160 TI - Molecular dynamics investigations of the polysaccharide scleroglucan: first study on the triple helix structure. AB - Explicit solvent molecular dynamics (MD) simulations on the triple helix of the polysaccharide Scleroglucan (Sclg) at two temperatures (273 and 300 K) were carried out. Owing to the complexity of the system, a united-atom force field, based on the properly modified GROMACS parameters, was adopted. To test these parameters for our system, MD simulations of the two disaccharidic units, representing the main chain and the side-chain linkages of the Sclg repeating unit, were performed and the results were compared with the literature data. The simulated triple helix of Sclg retained the main experimentally determined features of the polymer. The residence times of the solvent molecules at 273 and 300 K were analyzed. The results show that the more internal water molecules, interacting with the core of the Sclg triplex are not influenced substantially by changing the temperature, on the contrary the water molecules, interacting with the side-chain glucose residues show more significant differences. These data suggest that the more external water molecules, interacting with the side chain, play a major role in the conformational transition experimentally observed at low temperature. PMID- 16043161 TI - Crystal structure and aqueous solubility of ammonium D-glucarate. AB - Ammonium D-glucarate, NH(4)(C(6)H(9)O(8)) [ammonium D-saccharate, NH(4)-SAC], has been synthesized, and its crystal structure solved by single-crystal X-ray diffraction methods. NH(4)-SAC crystallizes in the monoclinic space group P2(1) (#4) with cell parameters a = 4.8350(4) Angstroms, b = 11.0477(8) Angstroms, c = 16.7268(12) Angstroms, beta = 90.973(1) degrees, V = 894.34(12) Angstroms(3), Z = 3. The structure was refined by full-matrix least-squares on F(2) yielding final R-values (all data) R1 = 0.0353 and R(w)2 = 0.0870. The structure consists of alternating (NH(4))(+) and (C(6)H(11)O(6))(-) layers parallel to the bc plane. An extended network of N-H...O(SAC) and O(SAC)-H...O(SAC) hydrogen bonds provide the 3-D connectivity. The aqueous solubility (S(w)) has been shown to be pH independent at ambient conditions within the range 4.5 < pH < 10 with S(w) = 2.19 M/L, whose value is about a factor of two lower than that of the ammonium isosaccharate analogue. PMID- 16043162 TI - Role of sequence variations in the human ether-a-go-go-related gene (HERG, KCNH2) in the Brugada syndrome. AB - BACKGROUND: Brugada syndrome (BrS) is an inherited electrical disorder associated with a high incidence of sudden death. In a minority of patients, it has been linked to mutations in SCN5A, the gene encoding the pore-forming alpha-subunit of the cardiac Na(+) channel. Other causally related genes still await identification. We evaluated the role of HERG (KCNH2), which encodes the alpha subunit of the rapid delayed rectifier K(+) channel (I(Kr)), in BrS. METHODS AND RESULTS: In two unrelated SCN5A mutation-negative patients, different amino acid changes in the C-terminal domain of the HERG channel (G873S and N985S) were identified. Voltage-clamp experiments on transfected HEK-293 cells show that these changes increase I(Kr) density and cause a negative shift of voltage dependent inactivation, resulting in increased rectification. Action potential (AP) clamp experiments reveal increased transient HERG peak currents (I(peak)) during phase-0 and phase-1 of the ventricular AP, particularly at short cycle length. Computer simulations demonstrate that the increased I(peak) enhances the susceptibility to loss of the AP-dome typically in right ventricular subepicardial myocytes, thereby contributing to the BrS phenotype. CONCLUSION: Our study reveals a modulatory role of I(Kr) in BrS. These findings may provide better understanding of BrS and have implications for diagnosis and therapy. PMID- 16043163 TI - Involvement of BKCa alpha subunit tyrosine phosphorylation in vascular hyporesponsiveness of superior mesenteric artery following hemorrhagic shock in rats. AB - OBJECTIVE: Vascular hyporesponsiveness is a major complication following severe trauma and shock. It plays important roles in the development of shock and seriously interferes with the treatment of shock. The mechanism responsible for the occurrence of vascular hyporesponsiveness has not been fully understood. The purpose of this study was to determine whether the alpha subunit tyrosine sites of large conductance calcium-activated potassium channel (BK(Ca)) could be phosphorylated and whether the phosphorylation of BK(Ca) was closely associated with the activation of BK(Ca) and the development of vascular hyporesponsiveness following hemorrhagic shock in rats. METHODS: A hemorrhagic shock (30 mm Hg for 0.5, 2, 4 h) model of Wistar rats was established. Phosphorylation of tyrosine residues of the BK(Ca) alpha subunit from vascular smooth muscle cells (VSMC) in superior mesenteric arteries (SMA) was detected by immunoprecipitation and Western blotting. BK(Ca) activity was evaluated by cell-attached patch clamping. The vascular responsiveness of SMA to norepinephrine was measured with an isolated organ perfusion system. RESULTS: The level of BK(Ca) alpha subunit tyrosine phosphorylation was increased in a time-dependent manner following hemorrhagic shock, which was mediated by protein tyrosine kinases (PTK) and protein tyrosine phosphatases (PTP). The activation of VSMC BK(Ca) following hemorrhagic shock was inhibited by genistein (2x10(-5) mol/L), the permeable isoflavone PTK inhibitor, and was potentiated by the PTP inhibitor sodium orthovanadate (Na(3)VO(4), 10(-3) mol/L). The decreased vasoresponsiveness following hemorrhagic shock was partly restored by genistein (10(-5) mol/L) or by the BK(Ca)-selective inhibitor tetrabutylammonium chloride (0.1 mmol/L), while it was further decreased by Na(3)VO(4) (10(-5) mol/L). CONCLUSION: The tyrosine residues of BK(Ca) alpha subunit of SMA were phosphorylated following hemorrhagic shock, which was regulated by PTK and PTP and appeared to be related to the activation of BK(Ca) and the development of vascular hyporesponsiveness following hemorrhagic shock. PMID- 16043164 TI - Contemporaneous carrier-state of two or three "proatherosclerotic" variants of APOE, ICAM1, PPARA and PAI-1 genes differentiate CAD patients from healthy individuals. AB - BACKGROUND: Atherosclerosis is the most important cause of coronary artery disease (CAD). Genetic predisposition to CAD is related to polymorphisms of genes encoding products functionally involved in pathogenesis of atherosclerosis. Polymorphisms of genes participating in monocyte adhesion and diapedesis, lipid metabolism and fibrinolysis regulation may be partially responsible for this process. The aim of our study was to assess the polymorphic variants frequencies of ICAM1, APOE, PPARA and PAI-1 genes in CAD patients and healthy blood donors and to find specific arrangement of polymorphic variants which would differentiate both groups. METHODS: We studied 146 CAD patients and 121 healthy blood donors. Polymorphisms in analyzed genes were examined using PCR-RFLP analysis. RESULTS: We found significantly higher frequency of 5G allele of PAI-1 gene in patients than in control subjects (p = 0.038, OR = 1.44). We observed also a considerably higher frequency of contemporaneous carriers of two or three "proatherosclerotic" variants: 1) PPARA and PAI-1, 2) APOE and ICAM1 and 3) PPARA, ICAM1 and PAI-1 in CAD group comparing to control subjects. The number of "proatherosclerotic" variants carriers differentiate studied groups also independently of specific genotype arrangement. CONCLUSION: In conclusion, contemporaneous carrier-state of two or three polymorphic variants within analyzed genes is associated with CAD. PMID- 16043165 TI - The phosphatidylcholine/lysophosphatidylcholine ratio in human plasma is an indicator of the severity of rheumatoid arthritis: investigations by 31P NMR and MALDI-TOF MS. AB - OBJECTIVES: Lipid second messengers, e.g. lysophosphatidylcholines (LPC) are involved in the pathogenesis of inflammatory diseases, for instance, rheumatoid arthritis (RA). Unfortunately, the analysis of LPC in complex mixtures as present in body fluids is still challenging. DESIGN AND METHODS: Matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) was applied for phospholipid (PL) analysis of organic extracts of synovial fluids from patients with RA as well as the corresponding plasma. These data were compared with results obtained by high resolution 31P NMR spectroscopy. RESULTS: Synovial fluids may be replaced by plasma since the analysis of both body fluids gives very similar results. Patients undergoing treatment with TNF-alpha inhibitors (ADALIMUMAB (HUMIRA)) were examined in order to investigate whether the clinically-significant attenuation of disease activity is accompanied by changes of the PL composition of plasma. It will be shown that especially the PC/LPC ratios of plasma represent a reliable measure of inflammation and increase upon therapy. CONCLUSIONS: Since plasma samples are readily available, our approach might be useful to draw conclusions before puncture of the affected joints is necessary and the PC/LPC ratio detected in plasma may serve as an indicator of RA in early stages. PMID- 16043166 TI - Segmentation of object outlines into parts: a large-scale integrative study. AB - In this study, a large number of observers (N=201) were asked to segment a collection of outlines derived from line drawings of everyday objects (N=88). This data set was then used as a benchmark to evaluate current models of object segmentation. All of the previously proposed rules of segmentation were found supported in our results. For example, minima of curvature (i.e. locations along the contour where negative curvature takes an extreme value) were often used as segmentation points. The second point of a pair connected by a segmentation line often depended on more global shape characteristics such as proximity, collinearity, symmetry, and elongation. Based on these results, a framework is presented in which all of the previously proposed (and now empirically validated) segmentation rules or rules for part formation are integrated. PMID- 16043167 TI - When English proposes what Greek presupposes: the cross-linguistic encoding of motion events. AB - How do we talk about events we perceive? And how tight is the connection between linguistic and non-linguistic representations of events? To address these questions, we experimentally compared motion descriptions produced by children and adults in two typologically distinct languages, Greek and English. Our findings confirm a well-known asymmetry between the two languages, such that English speakers are overall more likely to include manner of motion information than Greek speakers. However, mention of manner of motion in Greek speakers' descriptions increases significantly when manner is not inferable; by contrast, inferability of manner has no measurable effect on motion descriptions in English, where manner is already preferentially encoded. These results show that speakers actively monitor aspects of event structure, which do not find their way into linguistic descriptions. We conclude that, in regard to the differential encoding of path and manner, which has sometimes been offered as a prime example of the effects of language encoding on non-linguistic thought, surface linguistic encoding neither faithfully represents nor strongly constrains our mental representation of events. PMID- 16043168 TI - Multi-stage mental process for economic choice in capuchins. AB - We studied economic choice behavior in capuchin monkeys by offering them to choose between two different foods available in variable amounts. When monkeys selected between familiar foods, their choice patterns were well-described in terms of relative value of the two foods. A leading view in economics and biology is that such behavior results from stimulus-response associations acquired through experience. According to this view, values are not psychologically real; they can only be defined a posteriori. One prediction of this associative model is that animals faced for the first time with a new pair of foods learn to choose between them gradually. We tested this prediction. Surprisingly, we find that monkeys choose as effectively between new pairs of foods as they choose between familiar pairs of foods. We therefore, propose a cognitive model in which economic choice results from a two-stage mental process of value-assignment and decision-making. In a follow-up experiment, we find that the relative value assigned to three foods in sessions in which we tested them against each other combine according to transitivity. PMID- 16043169 TI - The influence of visual experience on the ability to form spatial mental models based on route and survey descriptions. AB - The purpose of the present study is twofold: the first objective is to evaluate the importance of visual experience for the ability to form a spatial representation (spatial mental model) of fairly elaborate spatial descriptions. Secondly, we examine whether blind people exhibit the same preferences (i.e. level of performance on spatial tasks) as sighted people in processing the type of perspective that is employed in a spatial description. Early blind, late blind and sighted participants listened to a route and a survey description of two environments. Next, they had to execute a recognition/priming task, a bird flight distance comparison task, and a scale model task. Spatial priming and symbolic distance effects were found for all participants. These findings suggest that early and late blind people can form spatial mental models on the basis of route and survey descriptions. Interestingly, in contrast with sighted people, blind people performed better after listening to a route than a survey description, even when the spatial problems that has to be solved explicitly favor the survey description. It seems that people with active vision build up a spatial mental model more efficiently from a survey description, while people with only visual memories (late blind), similar to people with no visual memories (early blind), build up a spatial mental model more efficiently from a route description. PMID- 16043170 TI - Potential role of nitration and oxidation reactions in the effects of peroxynitrite on the function of beta-adrenoceptor sub-types in the rat. AB - This study examined the hemodynamic responses elicited by the beta-adrenoceptor agonist, isoproterenol (1 and 10 microg/kg, i.v.) before and after administration of (i) peroxynitrite (10 x 10 micromol/kg, i.v.), (ii) the thiol chelator, para hydroxymercurobenzoic acid (pHMBA, 75 micromol/kg, i.v.), and (iii) the electron acceptor, nitroblue tetrazolium (NBT, 10 micromol/kg, i.v.) in pentobarbital anesthetized rats. The tachycardia elicited by the lower dose of isoproterenol was diminished whereas the tachycardia elicited by the higher dose was not attenuated after administration of peroxynitrite. The falls in hindquarter and renal vascular resistances elicited by both doses of isoproterenol were substantially diminished whereas the isoproterenol-induced falls in mesenteric vascular resistance were not changed after administration of peroxynitrite. All of the isoproterenol-induced responses were markedly attenuated after administration of pHMBA or NBT. These findings suggest that the oxidation and/or nitration of beta-adrenoceptors impair the ability of isoproterenol to bind to and/or activate these G protein-coupled receptors. beta1-, beta2- and beta3 adrenoceptors contain extracellular cysteine residues susceptible to oxidation (i.e., disulfide-bridge formation) whereas only the beta1- and beta2 adrenoceptors contain extracellular tyrosine residues susceptible to nitration. These findings also suggest that sustained impairment of beta1- and beta2 adrenoceptor function by peroxynitrite is due to nitration of extracellular tyrosine residues in these receptors. By analogy, beta3-adrenoceptors may not be permanently affected by peroxynitrite because these receptors are devoid of extracellular tyrosine residues. PMID- 16043171 TI - Comparative effects of levobupivacaine and racemic bupivacaine on excitotoxic neuronal death in culture and N-methyl-D-aspartate-induced seizures in mice. AB - We compared the neurotoxic profile of racemic bupivacaine and levobupivacaine in: (i) a mouse model of N-methyl-D-aspartate (NMDA)-induced seizures and (ii) in an in vitro model of excitotoxic cell death. When used at high doses (36 mg/kg) both bupivacaine and levobupivacaine reduced the latency to NMDA-induced seizures and increased seizure severity. However, levobupivacaine-treated animals underwent less severe seizures as compared with bupivacaine-treated animals. Lower doses of levobupivacaine and bupivacaine had opposite effects on NMDA-induced seizures. At doses of 5 mg/kg, levobupivacaine increased the latency to partial seizures and prevented the occurrence of generalized seizures, whereas bupivacaine decreased the latency to partial seizures and did not influence the development of generalized seizures. In in vitro experiments, we exposed primary cultures of mouse cortical cells, containing both neurons and astrocytes, to 100 microM NMDA for 10 min for the induction of excitotoxic neuronal death. This treatment killed 70-80% of the neuronal population, as assessed 24 h after the excitotoxic pulse. In this particular model, both levobupivacaine and bupivacaine were neuroprotective against NMDA toxicity. However, neuroprotection by levobupivacaine was seen at lower concentrations (with respect to bupivacaine) and was maintained at concentrations of 3 mM, which are much higher than the plasma security threshold for the drug in vivo. In contrast, no protection against NMDA toxicity was detected when 3 mM concentrations of bupivacaine were applied to the cultures. Our data show a better neurotoxic profile of levobupivacaine as compared to racemic bupivacaine, and are indicative of a safer profile of levobupivacaine in clinical practice. PMID- 16043172 TI - Novel (E)-alpha-[(1H-indol-3-yl)methylene]benzeneacetic acids as endothelin receptor ligands. AB - Endothelin-1 (ET-1), a peptide of 21 amino acid residues, is the most potent vasoconstrictor substance known and now it is understood to be one of a family of three mammalian vasoactive peptides that also includes ET-2 and ET-3. The endothelins (ETs) affect multiple organ systems and seem to be involved in the pathogenesis of many diseases such as hypertension, pulmonary hypertension, atherosclerosis, apoptosis inhibition and angiogenesis. The ETs exert their effects via activation of two distinct G-protein coupled receptor subtypes termed ET(A) and ET(B). To date a number of ET receptor ligands with good affinity and selectivity is known, nevertheless these compounds belong only to few chemical classes. The aim of this work was the identification of a "hit compound" with novel chemical structure, endowed with reasonable ET affinity and selectivity. Accordingly, a new class of (E)-alpha-[(1H-indol-3-yl)methylene]benzeneacetic acid derivatives (1-23) was synthesized for evaluation of their binding profiles. PMID- 16043173 TI - A method for the determination of minoxidil in hair-regrowth formulations by micellar electrokinetic capillary chromatography. AB - A method based on micellar electrokinetic capillary chromatography (MEKC) was developed for determination of minoxidil in Rogaine and competing products. The original intent of the work was to offer an orthogonal means to HPLC for testing illicit imitations of Rogaine. However, because the patent has since expired, we offer the procedure as a confirmatory measure to HPLC for assay of generic minoxidil products. The MEKC procedure complements an earlier method based on free solution capillary electrophoresis (FSCE), designed to the same end. Validation was carried out on both a Dionex CES-1, which utilizes gravity injection, and a PE-ABI 270HT, which employs vacuum injection. The procedure was validated for both active pharmaceutical ingredient and for minoxidil solutions. The run buffer is pH 7.0, 20 mM sodium phosphate, 20 mM sodium dodecyl sulfate, with 10% isopropanol; the internal standard is dl-tryptophan. The method bears the attributes of simplicity, ease of use, and short analysis time (12 min). It is selective with respect to known process and degradation impurities. High efficiency was achieved on the CES-1, with a plate count exceeding 200,000 for minoxidil at an elution time of 9 min. Although slight differences in performance were noted across the two instruments, results on both were in conformance with modern day validation expectations. Comparison of MEKC with HPLC resulted in slightly higher values for the former, but all results met registration specifications and internal targets. PMID- 16043174 TI - Diurnal nonstress test variations in the human fetus at risk. AB - OBJECTIVE: To compare the results of the nonstress test (NST) performed at 9:00 PM and 9:00 AM on women with high-risk pregnancies. METHOD: The NST was performed 2 h after a meal, at 9:00 AM and 9:00 PM, in a quiet room exposed to daylight, on 80 women with high-risk singleton pregnancies. Each session lasted 20 min. If the NST was nonreactive, the entire biophysical profile was immediately performed. The women's blood pressure was measured before, 10 min within, and at the end of the NST. Women who smoked or had uterine contractions were excluded from the study. RESULTS: Diurnal nonstress test variations were manifested by a higher incidence of reactive NSTs and an increased number of fetal heart accelerations after 9:00 PM (82.5%) than at 9:00 AM (68.8%) (p<0.027 P<0.001). CONCLUSIONS: Evening appointments for fetal assessments, except in emergency conditions, may eliminate the need for additional tests such as the entire biophysical profile because of the decreased incidence of nonreactive NSTs. Evening NSTs would save time and decrease maternal anxiety. PMID- 16043175 TI - Clinical experience and acceptability of the etonogestrel subdermal contraceptive implant. AB - OBJECTIVE: To evaluate efficacy, adverse effects, and user continuation rate of an etonogestrel subdermal single-rod contraceptive implant. METHODS: A total of 417 healthy volunteers of childbearing age were included in this multicenter trial. After implant insertion, the women were followed up during the 3 years of contraceptive action. At each visit, clinical findings, side effects, and bleeding patterns were recorded. Efficacy and continuation rates were analyzed using the Pearl Index and Kaplan-Meier life tables, respectively. RESULTS: The observation period totaled 958.5 woman-years (27.5 months per woman). The Pearl Index score was 0. Side effects were reported by 44.4% of users, but the proportion had decreased to 16.5% by the end of the study. The continuation rate was 61.4%. The most common reason for early discontinuation (in 21.1% of the participants) was menstrual disturbances. CONCLUSIONS: Etonogestrel subdermal contraceptive implants demonstrated high efficacy and an acceptable continuation rate. Counseling potential users explicitly about the side effects will optimize patient success with this long-acting contraceptive. PMID- 16043176 TI - Human papillomavirus (HPV) DNA detection in self-collected urine. AB - OBJECTIVE: Non-invasive sampling of human genitals to identify high-risk individuals with subclinical oncogenic HPV infection remains a challenge. The study was designed to see if self-collected urine can be used as a simple, non invasive sampling for screening HPV, particularly for screening/monitoring general population or young adolescents or infants, if they are to be immunized by HPV vaccines. METHOD: Self-collected urine samples from 100 sexually unexposed college going girls and cervical scrapes from 104 normal healthy sexually active married women were used in this study. Additionally, a group of 55 women were recruited for collecting first urine and later scraped cervical cells to validate urine sampling by directly comparing HPV positivity between the two types of biological specimens. A dry 'paper smear' method for specimen collection and a simple single tube protocol was employed for PCR detection of HPV infection. RESULTS: Out of 100 sexually inexperienced college going girls, only 6 (6%) were positive for HPV infection as revealed by L1 consensus primer and 4 (4%) of them were positive for HPV 16 but none was found positive for HPV 18 DNA. Out of 104 sexually active married women who were cytologically reported as negative by Pap test, 11 (10.5%) were found HPV positive and 7 (6.7%) of them had infection of high-risk HPV type 16. Both urine and later cervical scrapes from a group of 55 women collected as dry 'paper smear' showed perfect matching positivity for HPV between urine and cervical scrape. CONCLUSIONS: The use of urine coupled with its dry collection as 'paper smear' facilitating their easy transport, storage and direct PCR detection of HPV DNA opens up an alternative non-invasive approach for population screening of HPV infection, at least in such cases as children and infants in whom invasive samples are difficult to obtain. PMID- 16043177 TI - Matrix metalloproteinase 9 and relapse in patients with early stage squamous cervical carcinoma. PMID- 16043178 TI - Atosiban and nifedipin for the treatment of preterm labor. AB - OBJECTIVE: To perform a comparison between atosiban (oxytocin antagonist) and nifedipin (calcium channel blocker) for acute treatment of preterm labor and their maternal safety. METHODS: A randomized controlled trial study was performed on 80 pregnant women with preterm labor, between 26 and 34 weeks of pregnancy, in Akbar Abadi Teaching Hospital in Tehran, Iran. 40 women (the atosiban group) were compared with another 40 women (the nifedipin group) for the drugs' efficacy in delaying delivery for more than 48 h in order to undergo steroid therapy, and for more than 7 days or more, and also to assess their maternal safety. The duration between the drugs' administration and delivery were compared. The statistical analysis was performed using the Statistical Package for Social Science (SPSS). RESULTS: There was no statistically significant difference between the two groups in the treatment of preterm labor. Atosiban was effective in 82.5% of cases, and nifedipin in 75% of the cases (p=1.000), for delaying delivery for 48 h. Atosiban was effective in 75% of the cases, and nifedipin in 65% of the cases, for delaying delivery for more than 7 days. The maternal side effects in the atosiban group were 17.5%, and in the nifedipin group they were 40%, which had a statistically significant difference (p=0.027). The duration between treatment and delivery was 29.03+/-16.12 days in the atosiban group and 22.85+/-13.9 days in the nifedipin group with no statistically significant difference (p=0.79). CONCLUSION: Atosiban is an effective and safe drug for the acute treatment of preterm labor with minimal side effects, and it can be an option in the treatment of preterm labor, especially in patients with heart disease and multi-fetal pregnancies. PMID- 16043179 TI - Inflammatory bowel disease and preterm delivery. AB - OBJECTIVE: The present study investigates pregnancy outcome in women with IBD and examines the effect of pregnancy on the severity of IBD. METHOD: A case-control study comparing deliveries by mothers with IBD between January 1988 and January 2005 was performed. For every birth by a mother with IBD, four births by non-IBD mothers were randomly selected and adjusted for ethnicity and year of delivery. RESULT: During the study period there were 48 deliveries to patients with Crohn's disease and 79 deliveries to patients with ulcerative colitis. Higher rates of preterm delivery (<37 weeks) were found among patients with IBD as compared to the controls (odds ratios (OR)=2.2; 95% confidence interval (CI)=1.3-3.8). This association remained significant after adjustment for labor induction and multiple gestations, using the Mantel-Haenszel technique (weighted OR=2.1; 95% CI 1.3-3.5 and weighted OR=2.0; 95% CI 1.2-3.5; P=0.012; respectively). In addition, these patients had higher rates of fertility treatments (OR=2.2; 95% CI=1.1-4.4). Using a multivariate analysis, controlling for maternal age and fertility treatments, preterm delivery was seen to be significantly associated with IBD (adjusted OR=2.0; 95% CI=1.2-3.5). Perinatal outcomes, such as perinatal mortality, low Apgar scores, and congenital malformations, were comparable to the outcomes in the control group. CONCLUSION: Maternal IBD is an independent risk factor for preterm delivery. IBD is not associated with adverse perinatal outcome. PMID- 16043180 TI - Microwave endometrial ablation for menorrhagia in patients with systemic disorders. AB - OBJECTIVE: To evaluate the effectiveness of microwave endometrial ablation (MEA) in the treatment of menorrhagia in patients with severe systemic disease or medical conditions. METHODS: Forty-two menorrhagic women undergoing systemic disorders with failure of medical management were treated with MEA under local or general anesthesia, and were followed-up for 1 year. RESULTS: The women had a mean age of 39.4 years (range, 17-49). The procedure was successfully completed in all patients, and no intraoperative complications occurred. Two cases died of their primary severe medical diseases within 2 months of treatment but these cases were not associated with MEA. Among the remaining 40 patients, 24 (60.0%) had amenorrhea within 12 months. The duration of hospitalization and the amount of blood transfusion were significantly reduced after treatment, and the quality of life of these patients was improved significantly. CONCLUSIONS: MEA is a safe and effective treatment for the management of severe menorrhagia in patients undergoing systemic illness or severe medical conditions. PMID- 16043181 TI - Intravenous iron sucrose complex vs. oral ferrous sulfate for postpartum iron deficiency anemia. PMID- 16043182 TI - Pregnancy in patients with previous successful renal transplantation. AB - OBJECTIVE: To evaluate the risk factors affecting pregnancy, perinatal outcomes, and short-term graft condition in women who underwent renal transplantation. METHOD: Between May 1998 and January 2005, the histories of 20 pregnancies in 17 renal transplant recipients were reviewed retrospectively at the Ministry of Health Aegean Obstetrics and Gynecology Teaching Hospital. RESULT: There were significant associations between high serum creatinine level (>1.5 mg/dL) prior to pregnancy and preterm delivery (P=0.04), and between short interval between transplantation and pregnancy (<2 years) and increased rate of cesarean sections (P=0.04). There were no significant changes in serum creatinine levels during pregnancy in these women, and there were no acute rejection and graft loss during pregnancy or in the 6 months following delivery. CONCLUSION: These findings suggest that, although pregnancy does not adversely affect short-term renal allograft function, the rates of obstetric and perinatal complications are increased. Risk factors present before conception are a short interval between renal transplantation and pregnancy and poor renal function. PMID- 16043183 TI - Digital camera assessment for detection of cervical intraepithelial neoplasia in rural El Salvador. AB - OBJECTIVE: To explore the feasibility of digital photography for primary cervical cancer screening in a low-resource setting in El Salvador. METHODS: Three independent examiners performed Pap test, visual inspection, digital camera assessment and colposcopy on each subject. RESULTS: Lesions were detected in 99 of 504 patients (20%) by visual inspection, 72/504 (14%) by DART and 90/504 (18%) by colposcopic impression. Seven of 504 patients (1.3%) had CIN on histology. Pap detected 2 of 7 subjects (29% sensitivity) (C.I. 4%, 56%), visual inspection detected 5 of 7 (71% sensitivity, C.I. 34%, 95%), digital assessment detected 6 of 7 (86% sensitivity C. I. 45%, 99%), and colposcopic impression detected 5 of 7 (71% sensitivity, C.I. 34%, 95%). CONCLUSION: This small pilot trial demonstrates the potential value and feasibility of performing digital camera assessment of the reproductive tract on women in a developing country setting. PMID- 16043184 TI - Witnessed resuscitation: a concept analysis. AB - The science and practice of resuscitation is recognised and endorsed on an international level, yet for more than a decade it has appeared in the literature alongside words such as witnessing or witnessed to signify the practice of family presence during a resuscitation attempt. This paper explores the meaning of witnessed resuscitation using the process for concept analysis proposed by Rodgers. The term resuscitation is explored, followed by identification of relevant uses of the concept of witnessed resuscitation. The reader is introduced to conceptual variations that challenge the way in which the concept has become associated with family or relatives presence in the resuscitation room of an accident and emergency department. Conceptual clarity is further enhanced through the identification of references, antecedents and consequences of witnessed resuscitation and by providing a model case of the concept that includes its defining attributes. PMID- 16043185 TI - The effect of ezetimibe on serum lipids and lipoproteins in patients with homozygous familial hypercholesterolemia undergoing LDL-apheresis therapy. AB - LDL-apheresis is now commonly used as the only practical treatment for homozygous familial hypercholestreolemia (homozygous FH). However, even when applying apheresis therapy, the use of a drug or drugs is recommended to suppress the rapid rebound of cholesterol, which usually takes place after each apheresis procedure, and keep the LDL-cholesterol level within or near the optimal range for as long as possible. In this study, the usefulness of ezetimibe, a novel cholesterol-lowering drug, in enhancing the efficacy of apheresis therapy was evaluated in six Japanese patients with homozygous FH undergoing LDL-apheresis in combination with atorvastatin or simvastatin. With the exception of one patient, significant decreases in LDL-cholesterol at 2 weeks after each apheresis procedure were obtained during the period from 4 to 12 weeks of treatment, with an average reduction rate of 9.0% and a range of 4.3-12.6%. This corresponds to a suppression of rebound by approximately 36 mg/dl, from 391 to 355 mg/dl on average, in LDL-cholesterol values. Although the effect is not very strong, ezetimibe nevertheless appears to be a useful drug in combination with statins for those with homozygous FH undergoing LDL-apheresis. PMID- 16043186 TI - Guidance of engineered tissue collagen orientation by large-scale scaffold microstructures. AB - The tensile strength and stiffness of load-bearing soft tissues are dominated by their collagen fiber orientation. While microgrooved substrates have demonstrated a capacity to orient cells and collagen in monolayer tissue culture, tissue engineering (TE) scaffolds are structurally distinct in that they consist of a three-dimensional (3-D) open pore network. It is thus unclear how the geometry of these open pores might influence cell and collagen orientation. In the current study we developed an in vitro model system for quantifying the capacity of large scale ( approximately 200 microm), geometrically well-defined open pores to guide cell and collagen orientation in engineered tissues. Non-degradable scaffolds exhibiting a grid of 200 microm wide rectangular pores (1:1, 2:1, 5:1, and 10:1 aspect ratios) were fabricated from a transparent epoxy resin via high-resolution stereolithography. The scaffolds (n=6 per aspect ratio) were surface modified to support cell adhesion by covalently grafting GRGDS peptides, sterilized, and seeded with neonatal rat skin fibroblasts. Following 4 weeks of static incubation, the resultant collagen orientation was assessed quantitatively by small angle light scattering (SALS), and cell orientation was evaluated by laser confocal and scanning electron microscopy. Cells adhered to the struts of the pores and proceeded to span the pores in a generally circumferential pattern. While the cell and collagen orientations within 1:1 aspect ratio pores were effectively random, higher aspect ratio rectangular pores exhibited a significant capacity to guide global cell and collagen orientation. Preferential alignment parallel to the long strut axis and decreased spatial variability were observed to occur with increasing pore aspect ratio. Intra-pore variability depended in part on the spatial uniformity of cell attachment around the perimeter of each pore achieved during seeding. Evaluation of diamond-shaped pores [Sacks, M.S. et al., 1997. J. Biomech. Eng. 119(1), 124-127] suggests that they are less sensitive to initial conditions of cell attachment than rectangular pores, and thus more effective in guiding engineered tissue cell and collagen orientation. PMID- 16043187 TI - Sequential heavy metals extraction from polluted solids: influence of sulfate overconcentration. AB - The effect of sulfate on the chemical partitioning of Cu, Cd, and Pb in solid phases was assessed in this study. Modified BCR sequential extraction, speeded up by focused ultrasound, was systematically applied to various mixtures of typical geochemical solid phases (an artificial goethite spiked with Cu, Cd, and Pb and natural clays), with or without the addition of calcium sulfate. Sulfate was added so that three different concentrations were found in sequential extracts: 0.5, 1, and 1.5 g/L of sulfate. First, the results suggested that the goethite surface adsorption sites for sulfate are limited. Then, significant changes in Cd and Pb fractionation were observed in the presence of sulfate, whereas Cu remained strongly adsorbed on the solid phases. The main modifications observed in all the studied samples were a decrease in metal amounts in the first three fractions to the profit of an increase in the residual fraction. These results suggested that the adsorption of metals onto the studied solids was enhanced by the presence of sulfate. From these considerations, some hypotheses are advanced to describe the behavior of Cu, Cd, and Pb and their adsorption mechanisms on solid phases in sulfate-rich systems. PMID- 16043188 TI - Apparent and partial specific adsorption of 1,10-phenanthroline on mixtures of Ca montmorillonite, activated carbon, and silica gel. AB - The process of 1,10-phenanthroline adsorption at pH 5 on Ca-montmorillonite, activated carbon, and silica gel mixtures was studied as a function of the equilibrium concentration and the composition of the mixture. A model is presented for determining adsorption of the main component (the variable in the system) of the mixture, based on the thermodynamic concept of apparent and partial quantities, in combination with an equation representing total adsorption of the other two adsorbents as a function of the weight fraction of one of them and introducing the concept of mean total adsorption. The partial specific adsorption of orthophenanthroline (OP) on Ca-montmorillonite is strongly influenced by the presence of activated carbon and silica gel. Owing to a phenomenon of cationic exchange, adsorption on the clay is higher at low proportions in the mixture, but the strong effect of carbon and silica gel becomes apparent at increasing amounts of clay in the mixture. The partial specific adsorption of orthophenanthroline on activated carbon and silica gel was determined using a total adsorption equation for the two adsorbents as a function of the weight fraction of one of them and shows behavior inverse to that of adsorption on clay. PMID- 16043189 TI - Effect of temperature and salt on the phase behavior of nonionic and mixed nonionic-ionic microemulsions with fish-tail diagrams. AB - The phase behavior of Brij-56/1-butanol/n-heptane/water is investigated at 30 degrees C with alpha [weight fraction of oil in (oil+water)]=0.5, wherein a 2-->3 ->2 phase transition occurs with increasing W1 (weight fraction of 1-butanol in total amphiphile) at low X (weight fraction of both the amphiphiles in the mixture) and a 2-->1-->2 phase transition occurs at higher X. Addition of an ionic surfactant, sodium dodecylbenzene sulfonate, destroys the three-phase body and decreases the solubilization capacity of the system at different delta (weight fraction of ionic surfactant in total surfactant). A three-phase body appears at alpha=0.25, but not at alpha=0.75 for the single system. No three phase body appears with the mixed system at either alpha value. Increased temperature increases the solubilization capacity of the Brij-56 system; on the other hand, a negligible effect of temperature on the Brij-56/SDBS mixed system has been observed. Addition of salt (NaCl) produces a three-phase body for both single and mixed systems and increases their solubilization capacities. The monomeric solubility of 1-butanol in oil (S1) and at the interface (S1s) has been calculated using the equation hydrophile-lipophile balance plane for both singles and mixed-surfactant systems. These parameters have been utilized to explain the increase in solubilization capacity of these systems in the presence of NaCl. PMID- 16043190 TI - Determination of material models for arterial walls from uniaxial extension tests and histological structure. AB - An approach is proposed that allows the determination of material models from uniaxial tests and histostructural data including fiber orientation of the tissue. A combination of neo-Hookean and Fung-type strain-energy functions is utilized, and inequality constraints imposed on the constitutive parameters are derived providing strict local convexity and preferred fiber orientations. It is shown how the Fung-type model gets a pseudo-structural aspect inherent in the phenomenological model; a correlation between the fiber structure and the parameters of the Fung-type model is explicitly provided. In order to apply the proposed approach, quasi-static uniaxial extension tests of preconditioned prepared strips from the intima, media and adventitia of a human aorta with non atherosclerotic intimal thickening are acquired in axial and circumferential directions; structural information from histological analyses for each aortic tissue are documented. Data reveal a remarkable thickness, load-bearing capacity and stiffness of the intimal samples in comparison with the media and adventitia. Constitutive parameters for each aortic tissue layer are determined by solving the constrained problem using a penalty function method; a new approach for the estimation of appropriate start values is proposed. Finally, the predictivity and efficacy of the material models is shown by comparing model data with data from the uniaxial extension tests and histological image analyses. PMID- 16043191 TI - Why do dusk-active cockchafers detect polarization in the green? The polarization vision in Melolontha melolontha is tuned to the high polarized intensity of downwelling light under canopies during sunset. AB - In the retina of dusk-active European cockchafers, Melolontha melolontha, the linear polarization of downwelling light (skylight or light from the tree canopy) is detected by photoreceptors in upward-pointing ommatidia with maximal sensitivity at 520 nm in the green portion of the spectrum. To date no attempt has been made to answer the question of why these beetles detect polarization in the green. Here we present an atmospheric optical and receptor-physiological explanation of why longer wavelengths are advantageous for the perception of polarization of downwelling light under canopies illuminated by the setting sun. Our explanation focuses on illumination situations during sunset in canopied optical environments, because cockchafers are active at sunset and fly predominantly under canopies during their swarming, feeding, and mating periods. Using three simple atmospheric optical models, we computed the degree of linear polarization, the linearly polarized intensity of downwelling light, the quantum catch, and quantum catch difference between polarization detectors with orthogonal microvilli under canopies illuminated by the setting sun as functions of wavelength and solar zenith angle. Based upon these computations, we show that the green sensitivity of polarization detectors in M. melolontha is tuned to the high polarized intensity of downwelling light in the green under canopies during sunset, an optimal compromise between simultaneous maximization of the quantum catch and the quantum catch difference. We also briefly discuss how green sensitive polarization detectors can function efficiently enough during the pre feeding and egg-laying flights of cockchafers, which always occur prior to sunset and under the sky. PMID- 16043192 TI - A macrophage cell model for pH and volume regulation. AB - A whole-cell model of a macrophage (mphi) is developed to simulate pH and volume regulation during a NH4Cl prepulse challenge. The cell is assumed spherical, with a plasma membrane that separates the cytosolic and extracellular bathing media. The membrane contains background currents for Na+, K+ and Cl-, a Na(+)-K+ pump, a V-type H(+)-extruder (V-ATPase), and a leak pathway for NH4+. Cell volume is controlled by instantaneous osmotic balance between cytosolic and extracellular osmolytes. Simulations reveal that the mphi model can mimic alterations in measured pH(i) and cell volume (Vol(i)) data during and after delivery of an ammonia prepulse, which induces an acid load within the cell. Our analysis indicates that there are substantial problems in quantifying transporter-mediated H+ efflux solely from experimental observations of pH(i) recovery, as is commonly done in practice. Problems stemming from the separation of effects arise, since there is residual NH4+ dissociation to H+ inside the mphi during pH(i) recovery, as well as, proton extrusion via the V-ATPase. The core assumption of conventional measurement techniques used to estimate the H+ extrusion current (I(H)) is that the recovery phase is solely dependent on transporter-mediated H+ extrusion. However, our model predictions suggest that there are major problems in using this approach, due to the complex interactions between I(H), NH3/NH4+ buffering and NH3/NH4+ efflux during the active acid extrusion phase. That is, the conventional buffer capacity-based I(H) estimation must also take into account the perturbation that a prepulse challenge brings to the cytoplasmic acid buffer itself. The importance of this whole-cell model of mphipH(i) and volume regulation lies in its potential for extension to the characterization of several other types of non-excitable cells, such as the microglia (brain macrophage) and the T-lymphocyte. PMID- 16043193 TI - Using LogitBoost classifier to predict protein structural classes. AB - Prediction of protein classification is an important topic in molecular biology. This is because it is able to not only provide useful information from the viewpoint of structure itself, but also greatly stimulate the characterization of many other features of proteins that may be closely correlated with their biological functions. In this paper, the LogitBoost, one of the boosting algorithms developed recently, is introduced for predicting protein structural classes. It performs classification using a regression scheme as the base learner, which can handle multi-class problems and is particularly superior in coping with noisy data. It was demonstrated that the LogitBoost outperformed the support vector machines in predicting the structural classes for a given dataset, indicating that the new classifier is very promising. It is anticipated that the power in predicting protein structural classes as well as many other bio macromolecular attributes will be further strengthened if the LogitBoost and some other existing algorithms can be effectively complemented with each other. PMID- 16043194 TI - Physical exercise increases the expression of TNFalpha and GLUT 1 in muscle tissue of diabetes prone Psammomys obesus. AB - INTRODUCTION: Tumor necrosis factor-alpha (TNFalpha) is a major mediator of insulin resistance. On the other hand, it has been suggested that TNFalpha may facilitate glucose uptake through GLUT 1 expression. We recently found that physical exercise prevented the progression to type 2 diabetes mellitus in diabetes prone Psammomys obesus (sand rat). AIM: The aim of the present study was to characterize the influence of physical exercise on the expression of TNFalpha, its receptor R1 and GLUT 1 in muscle tissue of this animal model. METHODS: Animals were assigned for 4 weeks to four groups: high-energy diet (HC), high energy diet and exercise (HE), low-energy diet (LC), low-energy diet and exercise (LE). TNFalpha, R1 and GLUT 1 expression were analyzed using Western blot technique. RESULTS: None of the animals in the HE group became diabetic while all the animals in the HC group became diabetic. TNFalpha, its receptor (R1) and GLUT 1 expressions were significantly higher in the two exercising groups (LE and HE) and significantly lower in the HC group compared to the control LC group. CONCLUSIONS: Physical exercise augments the expression of TNFalpha, its receptor R1 and the glucose transporter GLUT 1 in muscle tissue. We suggest that this mechanism may improve glucose uptake through pathways parallel and unrelated to insulin signaling that may include MAPK and/or NO. These biochemical processes contribute to the beneficial effects of physical exercise on the prevention of type 2 diabetes mellitus. PMID- 16043195 TI - Life expectancy change in perturbed communities: derivation and qualitative analysis. AB - Pollution, loss of habitat, and climate change are introducing dramatic perturbations to natural communities and affecting public health. Populations in perturbed communities can change dynamically, in both abundance and age structure. While analysis of the community matrix can predict changes in population abundance arising from a sustained or press perturbation, perturbations also have the potential to modify life expectancy, which adds yet another means to falsify experimental hypotheses and to monitor management interventions in natural systems. In some instances, an input to a community will produce no change in the abundance of a population but create a major shift in its mean age. We present an analysis of change in both abundance and life expectancy, leading to a formal quantitative assessment as well as qualitative predictions, and illustrate the usefulness of the technique through general examples relating to vector-borne disease and fisheries. PMID- 16043196 TI - A comprehensive continuous-time model for the appearance of CGH signal due to chromosomal missegregations during mitosis. AB - Aneuploidy, the gain or loss of large regions of the genome, is a common feature in cancer cells. Irregularities in chromosomal copy number caused by missegregations of chromosomes during mitosis can be visualized by cytogenetic techniques including fluorescence in situ hybridization (FISH), spectral karyotyping (SKY) and comparative genomic hybridization (CGH). In the current work, we consider the propagation of irregular copy numbers throughout a cell population as the individual cells progress through ordinary mitotic cell cycles. We use an algebraic model to track the different copy numbers as states in a stochastic process, based on the model of chromosome instability of Gusev, Kagansky, and Dooley, and consider the average copy number of a particular chromosome within a cell population as a function of the cell division rate. We review a number of mathematical models for determining the length of the cell cycle, including the Smith-Martin transition probability model and the 'sloppy size' model of Wheals, Tyson and Diekmann. The program MITOSIM simulates the growth of a population of cells using the aforementioned models of the cell cycle. MITOSIM allows the cell population to grow, with occasional resampling, until the average copy number of a given chromosome in the population reaches a preset threshold signifying a positive copy number alteration in this region. MITOSIM calculates the relationship between the missegregation rate and the growth rate of the cell population. This allows the user to test hypotheses regarding the effect chromosomal aberrations have upon the cell cycle, cell growth rates, and time to population dominance. PMID- 16043197 TI - Genetic polymorphisms and possible gene-gene interactions in metabolic and DNA repair genes: effects on DNA damage. AB - We investigated in a central European population, the association between genetic polymorphisms in several genes coding for xenobiotic metabolizing enzymes (CYP1A1, CYP2E1, EPHX1, GSTP1, GSTM1 and GSTT1) and in DNA repair genes (XPD, XPG, XPC and XRCC1) and the levels of single-strand breaks (SSBs) and SSB endonuclease III sensitive sites (endoIII sites) in peripheral blood lymphocytes. No significant differences in the mean levels of SSBs and endoIII sites after stratification for main confounders and occupational exposure were observed in the studied population. Significantly higher levels of SSBs were observed in individuals bearing the wild-type alleles (AA) (0.75+/-0.51SSB/10(9)Da) and heterozygous (AC) genotypes (0.67+/-0.49SSB/10(9)Da) compared to those with homozygous XPD (CC) genotype (0.43+/-0.28SSB/10(9)Da, P=0.033). A moderate increase in the levels of SSBs was also found in individuals with the homozygous XPG exon 15 wild type (GG) and heterozygous (GC) genotypes in comparison to those with the homozygous (CC) genotype (P=0.066) and in individuals with low activity EPHX1 genotype in comparison to those with high activity genotype. Nevertheless, these differences were not statistically significant. No other significant association was found. When gene-gene interactions were evaluated, a combination of EPHX1 activity genotypes with that of either XPD or XPG significantly (P=0.003 and 0.016, respectively) modulated SSB levels resulting in a three-fold difference between the "protective" and the "adverse" genotype-combinations. Almost three-fold differences in SSB levels were found between the "protective" and the "adverse" genotype-combinations of EPHX1 activity genotype and GSTM1 or GSTT1 genotypes, respectively. In conclusion, our results suggest a relation between markers of genotoxicity and polymorphisms in genes coding for xenobiotic metabolizing and DNA repair enzymes as well as a modulating effect of combinations of these polymorphisms. PMID- 16043198 TI - The preferential mGlu2/3 receptor antagonist, LY341495, reduces the frequency of spike-wave discharges in the WAG/Rij rat model of absence epilepsy. AB - We examined the expression and function of group-II metabotropic glutamate (mGlu) receptors in an animal model of absence seizures using genetically epileptic WAG/Rij rats, which develop spontaneous non-convulsive seizures after 2-3 months of age. Six-month-old WAG/Rij rats showed an increased expression of mGlu2/3 receptors in the ventrolateral regions of the somatosensory cortex, ventrobasal thalamic nuclei, and hippocampus, but not in the reticular thalamic nucleus and in the corpus striatum, as assessed by immunohistochemistry and Western blotting. In contrast, mGlu2/3 receptor signalling was reduced in slices prepared from the somatosensory cortex of 6-month-old WAG/Rij rats, as assessed by the ability of the agonist, LY379268, to inhibit forskolin-stimulated cAMP formation. None of these changes was found in "pre-symptomatic" 2-month-old WAG/Rij rats. To examine whether pharmacological activation or inhibition of mGlu2/3 receptors affects absence seizures, we recorded spontaneous spike-wave discharges (SWDs) in 6-month old WAG/Rij rats systemically injected with saline, the mGlu2/3 receptor agonist LY379268 (0.33 or 1 mg/kg, i.p.), or with the preferential mGlu2/3 receptor antagonist, LY341495 (0.33, 1 or 5 mg/kg, i.p.). Injection of 1mg/kg of LY379268 (1 mg/kg, i.p.) increased the number of SWDs during 3-7 h post-treatment, whereas injection with LY341495 reduced the number of seizures in a dose-dependent manner. It can be concluded that mGlu2/3 receptors are involved in the generation of SWDs and that an upregulation of these receptors in the somatosensory cortex might be involved in the pathogenesis of absence epilepsy. PMID- 16043199 TI - Effect of chronic caffeine intake on choice reaction time, mood, and visual vigilance. AB - The stimulatory effects of acute caffeine intake on choice reaction time, mood state, and visual vigilance are well established. Little research exists, however, on the effects of chronic caffeine ingestion on psychomotor tasks. Therefore, the purpose of this study was to evaluate the effects of 5 days of controlled caffeine intake on cognitive and psychomotor performance. Three groups of 20 healthy males (age=22+/-3 years, mass=75.4+/-7.9 kg, body fat percentage=11.2+/-5.1%) twice completed a battery of cognitive and psychomotor tasks: after 6 days of 3 mg.kg(-1) day(-1) caffeine equilibration (Day 6), and after 5 days of experimental (0 [G0], 3 [G3], or 6 [G6] mg.kg(-1) day(-1)) caffeine intake (Day 11). Groups were randomized and stratified for age, mass, and body composition; all procedures were double-blind. Cognitive analyses involved a visual four-choice reaction time test, a mood state questionnaire, and a visual vigilance task. Experimental chronic caffeine intake did not significantly alter the number of correct responses or the mean latency of response for either the four-choice reaction time or the visual vigilance tasks. The Vigor-Activity subset of the mood state questionnaire was significantly greater in G3 than G0 or G6 on Day 11. All other mood constructs were unaffected by caffeine intake. In conclusion, few cognitive and psychomotor differences existed after 5 days of controlled caffeine ingestion between subjects consuming 0, 3, or 6 mg.kg(-1) day(-1) of caffeine, suggesting that chronic caffeine intake (1) has few perceptible effects on cognitive and psychomotor well-being and (2) may lead to a tolerance to some aspects of caffeine's acute effects. PMID- 16043200 TI - Aging effects on elevated plus maze behavior in spontaneously hypertensive, Wistar-Kyoto and Sprague-Dawley male and female rats. AB - Male and female spontaneously hypertensive (SHR), Wistar-Kyoto (WKY), and Sprague Dawley (SD) rats were assessed at one of two ages (postnatal day 74 or 346) for open field locomotor activity and anxiety-related behavior in the elevated plus maze (EPM). In general, the SHR displayed the least anxiety-related behavior, an effect that was magnified with age. At 11 months of age, the SHR more frequently entered and remained longer in the open arms than either the SD or the WKY strains. EPM behavior of the WKY strain was much less affected by age than that of the SD strain which displayed increased anxiety-related behavior with age. At the younger age, the typical sex effects were apparent; specifically, females exhibited a shorter duration in the closed arms. While the SHR were the most active strain in the EPM at both ages, they were more active in the open field only at the older age. In general, age-related changes in open field activity mirrored those of the EPM. These results provide a more comprehensive illustration of aging-related behavioral changes in male and female SHR, WKY and SD rats. PMID- 16043201 TI - Giant sperm cells with accessory macrotubules in a neuropteran insect. AB - The flagellar axoneme of the atypical spermatozoa (paraspermatozoa) of Mantispa perla (Neuroptera, Planipennia) contains accessory microtubules or rather macrotubules that are 55 nm in diameter and that has a wall consisting of about 40 protofilaments. The sperm tail further contains two giant mitochondrial derivatives, which during spermiogenesis store an electron dense material. The mature spermatozoon has a flattened acrosome and a elliptical nucleus. These giant spermatozoa may furnish nutrients to the functional spermatozoa (euspermatozoa) when they reach the female genital tracts or/and they function in sperm competition filling the spermatheca. PMID- 16043202 TI - P-glycoprotein activity and biological response. AB - P-glycoprotein (P-gp) is a transmembrane drug efflux pump encoded by the MDR-1 gene in humans. Most likely P-gp protects organs against endogenous and exogenous toxins by extruding toxic compounds such as chemotherapeutics and other drugs. Many drugs are substrates for P-gp. Since P-gp is also expressed in the blood brain barrier, P-gp substrates reach lower concentrations in the brain than in P gp-negative tissues. Failure of response to chemotherapy of malignancies can be due to intrinsic or acquired drug resistance. Many tumors are multidrug resistant (MDR); resistant to several structurally unrelated chemotherapeutic agents. Several mechanisms are involved in MDR of which P-gp is studied most extensively. P-gp extrudes drugs out of tumor cells resulting in decreased intracellular drug concentrations, leading to the MDR phenotype. Furthermore, the MDR-1 gene exhibits several single nucleotide polymorphisms, some of which result in different transport capabilities. P-gp functionality and the effect of P-gp modulation on the pharmacokinetics of novel and established drugs can be studied in vivo by positron emission tomography (PET) using carbon-11 and fluorine-18 labeled P-gp substrates and modulators. PET may demonstrate the consequences of genetic differences on tissue pharmacokinetics. Inhibitors such as calcium channel blockers (verapamil), cyclosporin A, ONT-093, and XR9576 can modulate the P-gp functionality. With PET the effect of P-gp modulation on the bioavailability of drugs can be investigated in humans in vivo. PET also allows the measurement of the efficacy of newly developed P-gp modulators. PMID- 16043203 TI - Indole-3-carbinol, but not its major digestive product 3,3'-diindolylmethane, induces reversible hepatocyte hypertrophy and cytochromes P450. AB - Indole-3-carbinol (I-3-C) and 3,3'-diindolylmethane (DIM) have been shown to reduce the incidence and multiplicity of cancers in laboratory animal models. Based on the observation that I-3-C induced hepatocyte hypertrophy when administered orally for 13 weeks to rats, a treatment and recovery study was undertaken to test the hypothesis that the induction of hepatocyte hypertrophy and cytochrome P450 (CYP) activity by I-3-C are adaptive, reversible responses. Additionally, we directly compared the effects of I-3-C to those of its principle metabolite DIM. Rats were treated orally for 28 days with 2 doses of I-3-C (5 and 50 mg I-3-C/kg body weight/day) and DIM (7.5 and 75 mg DIM/kg body weight/day) and then one-half of the animals were not treated for an additional 28 days. Organ weights, histopathology, and the CYP enzyme activities of 1A1/2, 2B1/2, 2C9, 2D6, 2E1, 3A4, and 19 A were measured both after treatment and after recovery. Oral administration of 50 mg I-3-C/kg body weight/day to rats for 28 days significantly increased liver weights and CYP enzyme activities. The effects in males were more pronounced and persistent after recovery than the effects in females. The increased organ weights returned to control values after treatment. Conversely, DIM did not alter liver weights and had no effect on CYP activities after the 28-day treatment. Some changes in CYP activities were measured after the DIM recovery period but the magnitudes of the changes were considered biologically insignificant. The results show that I-3-C, but not DIM, induces reversible adaptive responses in the liver. PMID- 16043204 TI - Long-term protection from SARS coronavirus infection conferred by a single immunization with an attenuated VSV-based vaccine. AB - Although the recent SARS coronavirus (SARS-CoV) that appeared in 2002 has now been contained, the possibility of re-emergence of SARS-CoV remains. Due to the threat of re-emergence, the overall fatality rate of approximately 10%, and the rapid dispersion of the virus via international travel, viable vaccine candidates providing protection from SARS are clearly needed. We developed an attenuated VSV recombinant (VSV-S) expressing the SARS coronavirus (SARS-CoV) spike (S) protein. In cells infected with this recombinant, S protein was synthesized, glycosylated at approximately 17 Asn residues, and transported via the Golgi to the cell surface. Mice vaccinated with VSV-S developed SARS-neutralizing antibody and were able to control a challenge with SARS-CoV performed at 1 month or 4 months after a single vaccination. We also demonstrated, by passive antibody transfer, that the antibody response induced by the vaccine was sufficient for controlling SARS CoV infection. A VSV-vectored SARS vaccine could have significant advantages over other SARS vaccine candidates described to date. PMID- 16043205 TI - Global gene expression analysis of two Streptococcus thermophilus bacteriophages using DNA microarray. AB - A custom microarray was developed to study the temporal gene expression of the two groups of phages infecting the Gram-positive lactic acid bacterium Streptococcus thermophilus. The complete genomic sequence of the virulent cos type phage DT1 (34,815 bp) and the pac-type phage 2972 (34,704 bp) were used for the construction of the microarray. Gene expression was measured at nine time intervals (0, 2, 7, 12, 17, 22, 27, 32 and 37 min) during phage infection and an expression curve was determined for each gene. Each phage gene was then classified into one of the three traditional transcription classes and these data were used to generate the complete transcriptional map of DT1 and 2972. Phage DT1 possesses 18 early genes, 12 middle genes and 12 late-expressed genes whereas 2972 has 16 early, 11 middle and 14 late genes. The trends of the phage gene expression profiles were also confirmed by slot blot hybridizations. Significant differences were observed when comparing the transcriptional maps of DT1 and 2972 with those already available for the S. thermophilus phages Sfi19 and Sfi21. To our knowledge, this report presents the first complete transcription analysis of bacteriophages infecting Gram-positive bacteria using the DNA microarray technology. PMID- 16043206 TI - Processivity factor of KSHV contains a nuclear localization signal and binding domains for transporting viral DNA polymerase into the nucleus. AB - Kaposi's sarcoma-associated human herpesvirus (KSHV) encodes a processivity factor (PF-8, ORF59) that forms homodimers and binds to viral DNA polymerase (Pol 8, ORF9). PF-8 is essential for stabilizing Pol-8 on template DNA so that Pol-8 can incorporate nucleotides continuously. Here, the intracellular interaction of these two viral proteins was examined by confocal immunofluorescence microscopy. When individually expressed, PF-8 was observed exclusively in the nucleus, whereas Pol-8 was found only in the cytoplasm. However, when co-expressed, Pol-8 was co-translocated with PF-8 into the nucleus. Mutational analysis revealed that PF-8 contains a nuclear localization signal (NLS) as well as domains located at the N-terminus and the C-proximal regions that are required for Pol-8 binding. This study suggests that the mechanism that enables PF-8 to transport Pol-8 into the nucleus is the first critical step required for Pol-8 and PF-8 to function processively in KSHV DNA synthesis. PMID- 16043207 TI - N(pro) of classical swine fever virus is an antagonist of double-stranded RNA mediated apoptosis and IFN-alpha/beta induction. AB - Classical swine fever virus (CSFV) protects cells from double-stranded (ds) RNA mediated apoptosis and IFN-alpha/beta induction. This phenotype is lost when CSFV lacks N(pro) (DeltaN(pro) CSFV). In the present study, we demonstrate that N(pro) counteracts dsRNA-mediated apoptosis and IFN-alpha/beta induction independently of other CSFV elements. For this purpose, we generated porcine SK-6 and PK-15 cell lines constitutively expressing N(pro) fused to the enhanced green fluorescent protein (EGFP). The survival of the SK6-EGFP-N(pro) cell line after polyinosinic polycytidylic acid [poly(IC)] treatment was comparable to that of CSFV-infected SK-6 cells and was significantly higher than the survival of the parent cell line. In PK-15 cells, the presence of EGFP-N(pro) prevented the DeltaN(pro) CSFV- and poly(IC)-mediated IFN-alpha/beta production. Importantly, N(pro) also inhibited IFN-alpha and IFN-beta promoter-driven luciferase expression in human cells and blocked IFN-alpha/beta induction mediated by Newcastle disease virus. This establishes a novel function for N(pro) in counteraction of the IFN-alpha/beta induction pathway. PMID- 16043208 TI - The recognition of partially visible natural objects in the presence and absence of their occluders. AB - The visual system is adept at compensating for the missing information in scenes that results from occlusion, but how this is done is not fully understood. In particular, the role of the occluding object in visual processing and its effect on the subsequent recognition of the occluded object is unclear. We report three human behavioral experiments suggesting that the recognition of partially visible objects is facilitated when the missing object information is replaced by an occluder rather than simply removed. Furthermore, we provide EEG evidence suggesting that the processes responsible for facilitated recognition occur relatively early in the visual stream. PMID- 16043209 TI - Lapse resistance in the verbal letter reporting task. AB - Lapses, or misreporting errors, can affect accuracy of threshold measurements. Assumptions about lapse rate, especially in untrained observers, have consequently guided the design of at least one clinical psychophysical test. Lapse rate was assessed using a verbal letter identification paradigm like that used in visual acuity and letter contrast sensitivity testing. Subjects occasionally made slip-of-the tongue errors but spontaneously corrected them. Lapse rate (excluding such errors) was 0-3 errors per 1,536 (average rate of 0.0005). In this common clinical paradigm, in which observers set their reporting pace, and where opportunity to amend responses is available, lapse rate is negligible. PMID- 16043210 TI - Removal of pharmaceuticals and fragrances in biological wastewater treatment. AB - The removal of seven pharmaceuticals and two fragrances in the biological units of various full-scale municipal wastewater treatment plants was studied. The observed removal of pharmaceuticals was mainly due to biological transformation and varied from insignificant (<10%, carbamazepine) to>90% (ibuprofen). However, no quantitative relationship between structure and activity can be set up for the biological transformation. Overall, it can be concluded that for compounds showing a sorption coefficient (K(d)) of below 300 L kg(-1), sorption onto secondary sludge is not relevant and their transformation can consequently be assessed simply by comparing influent and effluent concentrations. The two fragrances (HHCB, AHTN) studied were mainly removed by sorption onto sludge. For the compounds studied, comparable transformation and sorption was seen for different reactor types (conventional activated sludge, membrane bioreactor and fixed bed reactor) as well as for sludge ages between 10 and 60-80 days and temperatures between 12 degrees C and 21 degrees C. However, some significant variations in the observed removal currently lack an explanation. The observed incoming daily load of iopromide and roxithromycin in medium-sized municipal wastewater treatment plants (up to 80,000 population equivalents) is generated by only a small number of patients: the consequences for representative 24h composite sampling are discussed. Generally, the paper presents a method for setting up mass balances for micropollutants over entire wastewater treatment plants, including an estimation of the accuracy of the quantified fate (i.e. removal by sorption and biological transformation). PMID- 16043211 TI - Redox potential characterization and soil greenhouse gas concentration across a hydrological gradient in a Gulf coast forest. AB - Soil redox potential (Eh), concentrations of oxygen (O2) and three greenhouse gases (CO2, CH4, and N2O) were measured in the soil profile of a coastal forest at ridge, transition, and swamp across a hydrological gradient. The results delineated a distinct boundary in soil Eh and O2 concentration between the ridge and swamp with essentially no overlap between the two locations. Critical soil Eh to initiate significant CH4 production under this field conditions was about +300 mV, much higher than in the homogenous soils (about -150 mV). The strength of CH4 source to the atmosphere was strong for the swamp, minor for the transition, and negligible or even negative (consumption) for the ridge. Maximum N2O concentration in the soils was found at about Eh +250 mV, and the soil N2O emission was estimated to account for less than 4% for the ridge and transition, and almost negligible for the swamp in the cumulative global warming potential (GWP) of these three gases. The dynamic nature of this study site in response to water table fluctuations across a hydrological gradient makes it an ideal model of impact of future sea level rise to coastal ecosystems. Soil carbon (C) sequestration potential due to increasing soil water content upon sea level rise and subsidence in this coastal forest was likely limited and temporal, and at the expense of increasing soil CH4 production and emission. PMID- 16043212 TI - Pharmacology and pharmacogenetics of warfarin and other coumarins when used with supplements. PMID- 16043213 TI - Role of stretch-activated channels on the stretch-induced changes of rat atrial myocytes. AB - The role of stretch-activated channels (SACs) on the stretch-induced changes of rat atrial myocytes was studied using a computer model that incorporated various ion channels and transporters including SACs. A relationship between the extent of the stretch and the activation of SACs was formulated in the model based on experimental findings to reproduce changes in electrical activity and Ca(2+) transients by stretch. Action potentials (APs) were significantly changed by the activation of SACs in the model simulation. The duration of the APs decreased at the initial fast phase and increased at the late slow phase of repolarisation. The resting membrane potential was depolarised from -82 to -70 mV. The Ca(2+) transients were also affected. A prolonged activation of SACs in the model gradually increased the amplitude of the Ca(2+) transients. The removal of Ca((2+)) permeability through SACs, however, had little effect on the stretch induced changes in electrical activity and Ca(2+) transients in the control condition. In contrast, the removal of the Na(+) permeability nearly abolished these stretch-induced changes. Plotting the peaks of the Ca((2+)) transients during the activation of the SACs along a time axis revealed that they follow the time course of the Na(i)(+) concentration. The Ca((2+)) transients were not changed when the Na(i)(+) concentration was fixed to a control value (5.4mM). These results predicted by the model suggest that the influx of Na(+) rather than Ca(2+) through SACs is more crucial to the generation of stretch-induced changes in the electrical activity and associated Ca(2+) transients of rat atrial myocytes. PMID- 16043214 TI - The effects of Omega-3 fatty acids on growth regulation of epithelial ovarian cancer cell lines. AB - OBJECTIVE: Omega-3 fatty acids (OM-3FAs) have been shown to possess anti carcinogenic properties. We investigated the effect of OM-3FAs on epithelial ovarian cancer cell lines to determine if a growth suppressive effect existed and to gain a better insight on the potential molecular mechanisms that may be involved. METHODS: Three epithelial ovarian cancer cell lines (SKOV-3 [p53 null], TOV-21G [wt p53] and OVCAR-3 [mutant p53]) and one immortalized ovarian surface epithelial cell line (IOSE-29 [wt p53]) were treated with OM-3FAs and evaluated for cellular proliferation (WST-1 assay), apoptosis (Annexin V-FITC/PI method) and VEGF expression (VEGF ELISA assay). RESULTS: A statistically significant inhibitory effect under the influence of OM-3FAs was detected in all four cell lines. Apoptosis and VEGF down-regulation were either limited or not detected in the p53 null and mutant cell lines, SKOV-3 and OVCAR-3 respectively. Apoptosis and/or VEGF down-regulation was strongly evident in the wt p53 cell lines TOV-21G and IOSE-29. CONCLUSION: These data suggest that, under the influence of OM-3FAs, there are definitive growth suppressive mechanisms at work and that the biologic effects of OM-3FAs may in part be mediated by the p53 status. PMID- 16043215 TI - Impact of surgical approach on the management of macroscopic early ovarian borderline tumors. AB - OBJECTIVE: The aim of this study was to evaluate the impact of the surgical approach on the management and outcomes of patients with early borderline ovarian tumors (BOTs). MATERIAL AND METHODS: We retrospectively reviewed the medical charts of patients with stage Ia to Ic BOT treated surgically between January 1, 1985, and December 31, 2001. We compared patients initially managed by laparoscopy vs. laparotomy in terms of potentially harmful procedures and quality of staging. RESULTS: Of the 118 included patients, 48 (41%) had laparoscopy for initial surgery, 54 (45%) had laparotomy, and 16 (14%) had conversion from laparoscopy to laparotomy. Conservative treatment (57% of patients) was more common with laparoscopy (vs. laparotomy, P < 0.05) and in women older than 44 years (vs. younger than 44 years, P < 0.001). Intraoperative tumor rupture occurred in 9% of patients and was not associated with the surgical approach (P = 0.1). Bag extraction was used in 19 (40%) of the 48 laparoscopy patients. Staging was incomplete in 73% of patients overall. By univariate analysis, better quality of staging was associated with bilateral adnexectomy, age >44 years, laparotomy, hysterectomy, and treatment after 1995. By multivariate analysis, bilateral adnexectomy or hysterectomy was associated with better staging. Mean follow-up was 40 months, during which recurrence and survival rates were similar in the laparoscopy and laparotomy groups. CONCLUSION: Staging of macroscopic early stage BOTs was better in patients requiring radical surgery. After adjustment on disease severity, type of surgical access was not related to staging quality. PMID- 16043216 TI - Randomized controlled trial of a repeat mammography intervention: effect of adherence definitions on results. AB - BACKGROUND: As overall mammography rates approach national goals, mammography promotion efforts must increasingly focus on repeat adherence. This randomized controlled trial examined the effect of two interventions on repeat mammography utilization using various adherence definitions. METHODS: 1,558 women aged 40-63 receiving a mammogram through a federally funded screening program were randomized to three groups: mailed reminder (minimum group); mailed thank you card, newsletters, and reminder (maximum group); no mailings (control). The primary outcome (repeat mammogram) was assessed 13, 15, 18, and 24 months after the qualifying mammogram using administrative data. RESULTS: The proportions receiving a repeat mammogram within 13 months were 0.28, 0.30, and 0.32 for control, minimum, and maximum groups, respectively. The corresponding proportions were 0.38, 0.43, and 0.45 at 15 months; 0.43, 0.49, and 0.51 at 18 months; and 0.47, 0.52, and 0.54 at 24 months. There were no significant differences across study groups at 13 months. The differences between control and maximum subjects at 15, 18, and 24 months were statistically significant. The differences between control and minimum subjects were significant only at 18 months. CONCLUSIONS: The two low-cost mailed interventions evaluated modestly increased repeat mammography utilization. However, effects were not visible until at least 15 months after the qualifying mammogram. PMID- 16043217 TI - Damage mechanics and load failure of fiber-reinforced composite fixed partial dentures. AB - OBJECTIVES: Damage mechanics has been defined as the study of the initiation (initial failure) and accumulation of damage to and including rupture (final failure). This study was designed to evaluate the effect of increasing fiber reinforced composite (FRC) substructure within a standardized fixed partial denture (FPD) model on the failure performance, in terms of damage mechanics. METHODS: The two FRC restorative systems, Targis/Vectris (TV) (Ivoclar Vivadent) and EverStick (ES) (Stick Tech with Gradia, GC Corp.), were used to restore the molar FPD model (1.5 mm axial and 2.0 mm occlusal reduction). Templates were used to standardize substructure designs with 0, 18, 43, and 66% cross-sectional FRC volume fraction (V(FRC)) of fiber substructure. Specimens (n = 5) were homogenized at 29 points and stored for 1 week at 37 degrees C in distilled water. Specimens were luted with calcium hydroxide, then statically loaded until failure. Initial failure (IF), final failure (FF) and the mode of failure were recorded. RESULTS: The lowest mean load to initial failure was 530 N (TV 18%) and the highest was 1208 N (ES 66%). Linear regression analysis calculated the Pearson's correlation coefficient (r) for the interactions between V(FRC) and IF (ES: r = 0.7879, TV: r = 0.6184), V(FRC) and FF (ES: r = 0.912, TV: r = 0.8152), and between IF and FF (ES: r = 0.892, TV: r = 0.7237). Unreinforced specimens universally fractured instantaneously. SIGNIFICANCE: The highest loads to initial and final failure were yielded by specimens with the highest cross-sectional V(FRC). PMID- 16043218 TI - Pure red cell aplasia in B-cell lymphoproliferative disorder treated with rituximab: report of two cases and review of the literature. AB - Pure red cell aplasia (PRCA) is an unusual cause of anemia in patients with chronic lymphoproliferative disorders. Here, we present two cases of PRCA, one associated with chronic lymphocytic leukemia (CLL) and the other with splenic marginal zone lymphoma, in which the PRCA responded dramatically to treatment with rituximab. We then review the literature on PRCA in lymphoma and response to rituximab. PRCA associated with CLL or lymphoma may be another indication for rituximab therapy. PMID- 16043219 TI - 5-Aza-2'-deoxycytidine (decitabine) can relieve p21WAF1 repression in human acute myeloid leukemia by a mechanism involving release of histone deacetylase 1 (HDAC1) without requiring p21WAF1 promoter demethylation. AB - Decitabine is a potent demethylating agent that exhibits clinical activity against myeloid malignancies. Numerous genes silenced by hypermethylation are reactivated by decitabine through a mechanism involving promoter demethylation with subsequent release of histone deacetylases (HDACs) and accumulation of acetylated histones. Recent studies indicating that decitabine also induces regional chromatin remodeling of some unmethylated genes suggest additional mechanisms of action. Decitabine reactivates unmethylated p21WAF1 in some AML cell lines but the possible occurrence of p21WAF1 methylation in AML in vivo has not been studied in detail and decitabine effects on p21WAF1 chromatin remodeling have not been reported. We found that p21WAF1 mRNA was undetectable in 6 of 24 AML patient samples and 4 of 5 AML cell lines but there was no evidence of p21WAF1 promoter methylation. However, decitabine induced p21WAF1 in AML cell lines KG-1 and KG-1a in association with release of HDAC1 and increased acetylated histone H3 at the unmethylated p21WAF1 promoter. Decitabine effects on p21WAF1 histone acetylation and induction were enhanced by the HDAC inhibitor trichostatin A and were independent of wild type p53. Our findings indicate that decitabine can relieve p21WAF1 repression in AML by a mechanism that involves release of HDAC1 without requiring promoter demethylation. Furthermore, our study provides evidence that combined decitabine and HDAC inhibitor treatment can enhance chromatin remodeling and reactivation of an unmethylated tumor suppressor gene. This latter finding is of relevance to the clinical use of these agents in AML as we found the p21WAF1 promoter to be unmethylated in vivo. PMID- 16043220 TI - Recombinogenic activity of 10 chemical compounds in male germ cells of Drosophila melanogaster. AB - The Drosophila melanogaster male germ cells are sensitive to the recombinogenic activity of chemical compounds. In our experiments, we have employed four recessive markers located on the 2nd chromosome: dumpy (dp, 13.0), black (b, 48.5), cinnabar (cn, 57.5), and brown (bw, 104.5); b and cn flank the centromere. Three-day-old larvae, heterozygous for these markers, were treated chronically by oral administration with the test compounds. The chemicals already shown to be positive or negative in the assay systems to test chemical agents in D. melanogaster are three carcinogens (4-nitroquinoline N-oxide, hydroxylamine HCl, and acrylamide), four herbicides (maleic hydrazide, alachlor, trifluralin, and amitrol), and three insecticides (endrin, piperonyl butoxide, and allethrin). In our study, some compounds induced recombinogenic effects in Drosophila premeiotic male germ cells, and comparison of our results with those reported in the literature with the Drosophila wing somatic mutation and recombination assay showed that the somatic cells and the germinal cells have a differential response to the defined compounds. PMID- 16043221 TI - Investigation on the complex of diperoxovanadate with 2-(2'-pyridyl)-imidazole. AB - A novel diperoxovanadate complex NH4[OV(O2)2{2-(2'-pyridyl)-imidazole}] x 4H2O was synthesized in aqueous solution under physiological conditions. The solution structure of the complex was characterized by multinuclear (1H, 13C, 14N, and 51V) as well as multi-dimensional (DOSY and C-H COSY) NMR techniques in the interaction system of NH4VO3/H2O2/2-(2'-pyridyl)-imidazole at room temperature. The crystal structure of the complex was determined at 173K by single-crystal X ray diffraction method. It belongs to the monoclinic space group P21/c with a = 13.048(4), b = 6.984(2), c = 17.814(5) A, beta = 104.695(5), V = 1570.3(8) A3 and Z = 4. The crystal is composed of ammonium ions, {2-(2'-pyridyl) imidazole}oxodiperoxovanadate(V) ions, and water molecules, which are held together by ionic and hydrogen bond forces. The metal atom in the complex is seven-coordinated with a distorted pentagonal bipyramidal geometry. It is the first mononuclear diperoxovanadate complex with a N, N'-chelating biheteroaromatic ligand and its 51V chemical shift is at the highest field among the known mononuclear diperoxovanadate(V) complexes. PMID- 16043222 TI - Complexation of labile aluminium species by chelating resins Iontosorb--a new method for Al environmental risk assessment. AB - The utilization of chelating ion-exchange by the method based on binding strength and kinetic discrimination for aluminium fractionation was studied. Two chelating cellulose resins, Iontosorb Oxin (IO) and Iontosorb Salicyl (IS), were used for the determination of quickly reacting labile aluminium species. The possibilities of aluminium fractionation on these chelating resins were investigated by a solid phase extraction technique. The study of the pH (2.5-6.0) influence on the Al complexation by both resins indicates that at low pH the IS has lower sorption capacity but better adsorptive kinetic properties than IO. The optimal resin complexation time for reactive Al species was experimentally found after aluminium sorption study at pH 4.0 in synthetic solutions containing some inorganic and organic ligands, which simulate the composition of analysed acid soil and water samples. The negative influence of sulphate and iron on the Al complexation by IS resin was found and investigated. The flame atomic absorption spectrometry was used for the aluminium quantification. PMID- 16043223 TI - Kinetic studies of copper-induced oxidation of urate, ascorbate and their mixtures. AB - Urate and ascorbate are the major water-soluble low molecular weight antioxidants in serum. Much attention has been devoted to the effect of these antioxidants on lipoprotein peroxidation in vivo and on their effect on copper-induced peroxidation ex vivo. These studies revealed that urate inhibits ascorbate oxidation in vitro, whereas the effect of ascorbate on urate oxidation has not been systematically studied thus far. The present study addresses mechanistic aspects of the kinetics of copper-induced oxidation of both these antioxidants and their mutual effects in aqueous solutions. We found that: (i) ascorbate becomes oxidized much faster than urate. (ii) Urate inhibits the oxidation of ascorbate but, even in the presence of excess urate, ascorbate becomes oxidized much faster than urate. (iii) Ascorbate, as well as the products of its oxidation (and/or hydrolysis) inhibit the copper-induced oxidation of urate. All these results are consistent with the hypothesis that the rate of ascorbate oxidation is determined by the rate of reoxidation of reduced copper (Cu(I)) to Cu(II) by molecular oxygen, whereas the rate of urate oxidation is governed by the rate of oxidation of urate within a 2:1 urate/copper complex. We think that the mutual effects of urate and ascorbate on each other's oxidation are likely to enhance their inhibitory effect on lipid peroxidation in biologically relevant systems including membranes and lipoproteins. PMID- 16043224 TI - In vivo measurement of bone aluminium: recent developments. AB - A biomarker of aluminium accumulation in the human body can play a valuable role in determining health effects of chronic aluminium exposure, complementing other human and environmental monitoring data. In vivo neutron activation provides such a non-invasive biomarker. To date, the best in vivo neutron activation system used thermalised neutrons from a nuclear reactor at Brookhaven National Laboratory, which suffered only slightly from interference from other elements, primarily phosphorus, and from the disadvantage of restricted accessibility. At McMaster, we use a nuclear reaction on an accelerator to select neutron energy, which eliminates the interferences. Spectral decomposition analysis improved sensitivity. A new 4pi detection system also enhanced sensitivity. Together these improvements yield a minimum detection limit of 0.24 mgAl in a hand, slightly better than at Brookhaven and equivalent to "normal" levels. Further improvements should result from a new irradiation cavity and from using a higher proton current on the accelerator to shorten irradiation times. The system is now ready for pilot human studies. PMID- 16043225 TI - Congenital toxoplasmosis from an HIV-infected woman as a result of reactivation. AB - Congenital toxoplasmosis usually results from acquired infection in non-immune pregnant women. However, severely HIV-infected women with a latent Toxoplasma infection can transmit the parasite as a result of reactivation. We report a case of toxoplasmic reactivation in an HIV-infected woman with moderate immunosuppression resulting in a severe congenital toxoplasmosis. PMID- 16043226 TI - Comparative anatomical assessment of the piglet as a model for the developing human medullary serotonergic system. AB - Because the piglet is frequently used as a model for developmental disorders of the medullary serotonergic (5-HT) system in the human infant, this review compares the topography and developmental profile of selected 5-HT markers between humans in the first year of life and piglets in the first 60 days of life. The distribution of tryptophan hydroxylase-immunoreactive 5-HT neurons in the human infant medulla is very similar, but not identical, to that in the piglet. One notable difference is the presence of compact clusters of 5-HT neurons at the ventral surface of the piglet medulla. While it lacks these distinctive clusters, the human infant medulla contains potentially homologous 5 HT neurons scattered along the ventral surface embedded in the arcuate nucleus. Each species shows evidence of age-related changes in the 5-HT system, but the changes are different in nature; in the human infant, statistically significant age-related changes are observed in the proportional distribution of medullary 5 HT cells, while in the piglet, statistically significant age-related changes are observed in the levels of 5-HT receptor binding in certain medullary nuclei. Analyses of 5-HT receptor binding profiles in selected nuclei in the two species suggest that the equivalent postnatal ages for 5-HT development in piglets and human infants are, respectively, 4 days and 1 month, 12 days and 4 months, 30 days and 6 months, and 60 days and 12 months. Collectively, when certain species differences are considered, these data support the use of the piglet as a model for the human infant medullary 5-HT system. PMID- 16043227 TI - Microdialysis of proteins: performance of the CMA/20 probe. AB - Unlike all other currently available sampling techniques, microdialysis allows the continuous recovery of dialysed fluid from the interstitial space of living tissue. Microdialysis has been extensively used to study small molecules such as neurotransmitters, metabolites and drugs in the brain and other tissues. There has been increasing interest in measuring proteins using microdialysis. Optimizing protein recovery requires slow buffer flow rates, large pore membranes and osmotic balancing. An examination of a widely used commercially available large pore (100kDa MWCO) polymeric microdialysis probe, the CMA/20 (polyethersulfone) over 6 days of continuous microdialysis showed that: (1) published molecular weight cut-off values may not predict the size of proteins that can be recovered. This membrane had an effective molecular weight cut-off values (MWCO) of about 29kDa, (2) protein recoveries decrease over time, (3) small proteins are much less affected than larger ones and (4) there can be significant differences in the recovery of proteins similar in size perhaps due to reaching of a critical size, protein aggregation, shape, surface charge or hydrophobicity. PMID- 16043228 TI - A method for intracochlear drug delivery in the mouse. AB - The confluence of two rapidly emerging research arenas - development of mouse models of human deafness and inner ear drug therapy for treatment and prevention of hearing loss - provides an opportunity for unprecedented approaches to study and treat deafness. Toward such goals, we have developed a method for intracochlear drug delivery in the mouse. The bulla was exposed using a ventral approach and the stapedial artery cauterized. An opening made into the inferior medial aspect of the bulla, where the basal cochlear wall fuses with tympanic bulla, provided direct access to the scala tympani without separately opening the bulla or elevating auditory response thresholds. Cochlear responses, assayed by frequency-specific effects on ABRs and DPOAEs, were stable with infusion (1 microl/h) of an artificial perilymph solution (80 min). The glutamate receptor antagonist, CNQX (100 microM; 175 min), reduced ABR responses without affecting DPOAEs. Salicylate (5mM; 165 min) altered both. Both drugs had greatest effects at high frequencies, but distributed throughout the cochlea and were reversible. The safe delivery of drugs into the cochlea by this approach has immediate application in the study and treatment of various forms of human hearing loss that can be modeled in the mouse. PMID- 16043229 TI - Examining the association between pediatric bipolar disorder and anxiety disorders in psychiatrically referred children and adolescents. AB - BACKGROUND: To examine the association between anxiety (ANX) and bipolar disorders (BPD) in a clinically referred pediatric sample. METHODS: Subjects were consecutively referred youth aged <18 years, with a DSM-III-R/DSM-IV diagnosis of BPD (n=297) or a disruptive behavior disorder (DBD) (n=1100) on structured interview. RESULTS: Bipolar disorder in youth was associated with a significantly increased risk for most of the DSM-IV anxiety disorders and was not specifically linked to any one disorder in particular. CONCLUSIONS: Bipolar disorder in youth is a significant risk factor for anxiety comorbidity. Considering the morbidity and dysfunction as well as the differing therapeutic needs associated with bipolar and anxiety symptomatology, appropriate therapeutic approaches will be needed for the management of youth with this comorbidity. PMID- 16043230 TI - Tumor-necrosis-factor-related apoptosis-inducing-ligand (TRAIL)-mediated death of neurons in living human brain tissue is inhibited by flupirtine-maleate. AB - Neuronal damage mediated by the TRAIL-system might be involved in the pathogenesis of neuroinflammatory diseases of the central nervous system. Here we used an investigator-independent approach to quantify TRAIL-mediated death of total CNS cells and neurons in a living human brain slice culture system, a model which is much closer to the in vivo situation than dissociated cell culture. We observed dose-dependent TRAIL-mediated death of both total human CNS cells and neurons, which was prevented by flupirtine-maleate, a centrally acting analgesic drug with proposed neuroprotective properties. Our data suggest flupirtine maleate as an orally available neuroprotective approach in the course of neuroinflammation. PMID- 16043232 TI - Immediate repair of the tympanic membrane to prevent persistent perforation after intentional removal of long-lasting tubes. AB - OBJECTIVE: Our purpose was to evaluate efficiency of immediate repair of the tympanic membrane perforation after intentional removal of the long-lasting tubes. METHODS: This study was done in 36 ears of 27 patients who had either Goode-T or Paparella-II silicone tube insertion due to chronic OME or ROM and tube removal. Only the ears in which tube removal was performed due to no longer need for middle ear ventilation were included to the study. After removal of the tube, the perforation edges were refreshed and Steri-Strip (3M) patch was adhered on the perforation site. Otoscopic, tympanometric and audiologic data were reported. RESULTS: Mean duration of the tube persistence was 49.58+/-11.94 months. It was found that there were two subgroups in the study group: those under regular follow-up (20 ears), and the ears which were out of regular follow up (16 ears). Mean tube persistence times were 34.10 and 52.11 months in these subgroups, respectively (chi2-test, p=0.056). In six ears (16.67%), persistent perforation (PP) was found. PP rate (PPR) was higher in group-B (25%) than in group-A (10%) (chi2-test, p>0.1). The rest perforation was anteriorly marginal in five of six PP (83.33%). The PPR in the ears in which rest perforation was anteriorly marginal was 35.71% (5/14) while it was 4.54% (1/22) in the ears with central rest perforation (chi2-test, p<0.02). CONCLUSIONS: Even immediate repair of the perforation after removal of the long-lasting tubes resulted in a high PPR. The data in this study documented that this high PPR was associated with type and localisation of rest perforation and tube persistence time. Anteriorly, marginal perforations had about eight times higher risk of PP and longer tube persistence caused higher anterior marginal perforations after tube removal. PMID- 16043233 TI - Phenotypic characterization of a DFNA6 family showing progressive low-frequency sensorineural hearing impairment. AB - Only three autosomal dominant hearing loss loci (DFNA1, DFNA6/14/38 and DFNA54) have been reported to be associated with predominantly low-frequency (<2kHz) sensorineural hearing impairment (LFSNHI). The DFNA6 locus was previously mapped to chromosome 4p16.3. It was showed that WFS1 is located in this region. This study presents a six-generation family from Hungary with nonsyndromic, post lingual, bilateral, symmetric, progressive LFSNHI, that discloses positive linkage to the DFNA6 region. Eleven genetically affected family members have LFSNHI. The HI is started before the age of 25 years. The severity of HI varies from mild to moderate, related to age. Progression was mild but significant at all frequencies causing a flat type audiogram. High-resolution temporal bone CT scan showed normal external, middle and inner ear without any osseus malformations in the temporal bone. Studying genotype-phenotype correlations will enhance our understanding of normal and disturbed hearing process. PMID- 16043234 TI - Effect of age at cochlear implantation on open-set word recognition in Mandarin speaking deaf children. AB - OBJECTIVE: The purpose of this study was to determine whether age at cochlear implantation influences open-set speech perception in children after long-term use of the implant device. METHOD: Twenty-eight congenitally deafened children, receiving implants of Nucleus CI24M devices, were divided into two groups: (1) CI < 3: those who received implants before 3 years of age and (2) CI > 3: those who received implants after 3 years of age. We compared open-set speech perception in CI < 3 and CI > 3 after 4-5 years of device use. Speech perception tests were conducted using the Mandarin Lexical Neighborhood Test (M-LNT). Unpaired t-test was applied for statistical analysis, and p < 0.05 was considered significant. RESULTS: In CI < 3, the average of percent correct was 80.0 +/- 8.8 and 70.5 +/- 9.2% on, respectively, the easy and hard versions of the M-LNT. By contrast, in CI > 3, the average percent correct was 62.5 +/- 19.9 and 59.1 +/- 15.2%, respectively. Regardless of the M-LNT version used, CI < 3 performed significantly better than CI > 3 (easy, p = 0.005 versus hard, p = 0.022). CONCLUSION: The present investigation demonstrated that age at implantation influences open-set speech perception of cochlear implanted children 4-5 years after device connection. Implantation before 3 years of age promotes the development of open-set speech perception abilities in congenitally deafened children. PMID- 16043235 TI - Piperine: researchers discover new flavor in an ancient spice. AB - Studies with animals that are deficient in the vanilloid (capsaicin) receptor TRPV1 have confirmed the pivotal role that TRPV1 has in the development of post inflammatory hyperalgesia, and enhanced TRPV1 expression has been described in various human disorders. Natural products have provided several lead structures for the development of vanilloid ligands. A recent study shows that piperine, the irritant principle in black pepper, is more efficient than capsaicin in the desensitization of human TRPV1, which suggests that this pharmacological aspect of vanilloids can be dissociated from its potency. This finding raises the intriguing possibility that piperine can be used as a chemical template for the design of improved TRPV1 agonists. PMID- 16043236 TI - A cosmid-based system for inserting mutations and foreign genes into the simian varicella virus genome. AB - Simian varicella is a natural varicella-like disease of nonhuman primates. The etiologic agent, simian varicella virus (SVV), is genetically related to varicella-zoster virus (VZV) and SVV infection of nonhuman primates is a useful model to investigate VZV pathogenesis and latency. In this study, we report development of a cosmid-based genetic system to generate SVV mutant viruses. SVV subgenomic DNA fragments (32-38kb) that span the viral genome were cloned into cosmid vectors. Co-transfection of Vero cells with four overlapping cosmid clones representing the entire SVV genome resulted in recombination and generation of infectious virus. SVV mutants were produced by manipulation of one cosmid and substitution into the genetic system. This genetic approach was used to insert a site-specific mutation within the SVV open reading frame 14 which encodes the nonessential glycoprotein C gene. In a subsequent experiment, the green fluorescent protein (GFP) gene was inserted into the SVV genome within ORF 14. These SVV mutants replicate as efficiently as wild-type SVV in cell culture. This cosmid-based genetic system will be useful to investigate the effect of viral mutations on SVV pathogenesis and latency and also to develop and evaluate recombinant varicella vaccines that express foreign antigens. PMID- 16043237 TI - Human diseases reveal novel roles for neural laminins. AB - Extracellular matrix molecules such as laminins have a central role in regulating cell behaviour. However, our understanding of their functions in the mammalian nervous system is incomplete. It is important to establish these functions, both for an understanding of normal development and to devise strategies to enhance repair. Here, we review how insights gained from human diseases caused by genetic mutations in laminins or their receptors have revealed significant and sometimes unexpected roles for laminins in neural stem cells, migrating neurons and myelinating glia, in both the PNS and CNS. PMID- 16043238 TI - Shedding light on membrane proteins. AB - Membrane proteins are a cell's first line of communication with the world that exists just beyond the plasma membrane. These proteins afford the cell a peek at its external environment, signal the cell to adjust its internal chemistry in response to its surroundings, and ensure that the cell's metabolic state is faithfully coupled to the outside world. Because of their importance in cellular communication, membrane proteins have been the focus of intense study at the functional and structural levels. Here, we describe optical techniques that can either passively monitor or actively control the structural rearrangements that take place as these proteins peek at the outside world. Our focus is on ion channels, but the techniques described can be applied to a host of other proteins. PMID- 16043239 TI - Animal models of multiple system atrophy. AB - Multiple system atrophy (MSA) is a fatal neurodegenerative disorder presenting with autonomic failure and motor impairment, primarily comprising L-dopa resistant parkinsonism but occasionally involving cerebellar ataxia. These features result from progressive multisystem neuronal loss that is associated with oligodendroglial alpha-synuclein inclusions. The growing number of animal models for MSA reflects the search for a preclinical test-bed for elucidating MSA pathogenesis and for developing novel therapeutic interventions. Here, the currently available MSA animal models will be reviewed and leads for future research will be identified. PMID- 16043240 TI - Behavioral response to novelty correlates with dopamine receptor availability in striatum of Gottingen minipigs. AB - Behavioral response to novelty in rats has been linked both to dopamine transmission in the ventral striatum, and to propensity to self-administer psychostimulant drugs. In order to probe the relationship between behavioral response to novelty and dopamine systems we have developed a behavioral model for correlation with positron emission tomography (PET) of dopamine transmission in brain of Gottingen minipigs. In the present study, we measured exploration of a novel object by recording the number of contacts, and duration of contact with a novel object, in groups of six male and six female adult minipigs. We hypothesized that these novelty scores would correlate with the amphetamine evoked dopamine release in ventral striatum, measured 2 weeks later in a PET study of the availability of binding sites for the dopamine D2/3 antagonist [11C]raclopride. There were significant correlations between duration of contact with a novel object and the amphetamine-evoked reductions in binding potential (DeltapB) in the left ventral striatum of the 12 animals; Comparison of results by gender revealed that the correlation was driven mainly by the male group, and was not present in the female group. We interpret these results to show that propensity to explore an unfamiliar object is relatively elevated in pigs with low basal occupancy of dopamine D2/3 receptors by endogenous dopamine, and with high amphetamine-induced occupancy of released dopamine in the male pigs. PMID- 16043241 TI - Metabotropic glutamate receptor 1 blockade impairs acquisition and retention in a spatial Water maze task. AB - Metabotropic glutamate receptors, including the mGlu1 receptor, have received considerable attention as potential targets for anxiolytic, antidepressant, antipsychotic and antinociceptive drugs. mGlu1 receptors have also been suggested to play a role in the modulation of cognitive processes, but knowledge is still very limited. In the present study the effects of the selective mGlu1 receptor antagonist 3,4-dihydro-2H-pyrano[2,3]beta-quinolin-7-yl)(cis-4 methoxycyclohexyl)methanone (JNJ16259685, 0.63-10 mg/kg s.c.) on more or less spatially demanding learning and spatial memory (retention and re-acquisition) were investigated in mice performing in a water maze. Selective mGlu1 receptor blockade with JNJ16259685 impaired spatial acquisition processes, irrespective of spatial load, as well as spatial re-acquisition, already at the lowest dose tested (0.63 mg/kg). In contrast, effects on spatial retention performance were relatively mild in mice that had learned to locate the position of the escape platform prior to treatment. Thigmotaxic behaviour and locomotor activity appeared to be unaffected by JNJ16259685. These data suggest that blockade of the mGlu1 receptor primarily affects learning of new information, but leaves retention of spatial information relatively unaffected. Blockade of the mGlu5 receptor with MPEP also impaired spatial learning, although only at the highest dose tested (10 mg/kg). An ex vivo receptor occupancy study in rats revealed that MPEP occupied central mGlu5 receptors with an ED(50) of 2.0 mg/kg one hour after subcutaneous administration. This is 50-150 times higher than the ED(50) reported for JNJ16259685 at central mGlu1 receptors and suggests that one reason why the two compounds cause cognitive effects at different doses might be due to differences in central mGlu receptor occupancy, rather than fundamentally different roles of mGlu1 and mGlu5 receptors in the modulation of cognitive function. PMID- 16043242 TI - Genotoxic, cytotoxic, developmental and survival effects of tritiated water in the early life stages of the marine mollusc, Mytilus edulis. AB - Using an integrated approach linking different levels of biological organisation, the genotoxic, cytotoxic, developmental and survival impact of tritiated water (HTO) were investigated in the embryo-larvae of marine mollusc Mytilus edulis. One-hour-old embryos were exposed to a range of concentrations (0.37-370 kBq ml( 1)) of HTO, which delivered a dose between 0.02 and 21.41 mGy over the exposure period for different end points. Detrimental effects, if any, were monitored at different levels of biological organisation (i.e. DNA, chromosomal, cellular and individual). Genotoxic effects were assessed using molecular and cytogenetic approaches which included analysis of random amplified polymorphic DNA (RAPD), induction of sister chromatid exchanges (SCEs) and chromosomal aberrations (Cabs). Cytotoxic effects were evaluated by determining the proliferative rate index (PRI) of the embryo-larval cells. Developmental and survival effects were also monitored every 24 h up to 72 h. Results in general indicated that HTO significantly increased cytogenetic damage, cytotoxicity, developmental abnormalities and mortality of the embryo-larvae as a function of concentration or radiation dose. The analysis of RAPD profiles also revealed qualitative effects in the HTO exposed population compared to controls. However, while the embryo-larvae showed dose or concentration dependent effects for mortality, developmental abnormalities and induction of SCEs, the dose-dependent effects were not apparent for Cabs and PRI at higher doses. The study contributes to our limited understanding of the impact of environmentally relevant radionuclides on non-human biota and emphasises the need for further investigations to elucidate potentially long term damage induced by persistent, low levels of other radionuclides on commercially and ecologically important species, in order to protect human and ecosystem health. PMID- 16043243 TI - Expression of endothelin-1 system in a pig model of endotoxic shock. AB - Endothelin (ET)-1 is a potent vasoconstrictive peptide and it is involved in the pathogenesis of septic shock. Blockade of ET-1 receptors abolishes the LPS induced pulmonary hypertension and worsens the LPS-dependent systemic hypotension, but the role of ET-1 in sepsis remains uncertain. To determine the role of ET-1 in cardiovascular and respiratory derangement in a porcine model of endotoxemic shock we evaluated ET-1 plasma levels and ET-1 mRNA and protein levels in lung, liver, and heart as well as Endothelin Converting Enzyme-1, ET(A) and ET(B) receptors mRNA in the same tissues. Twelve piglets were randomised to sham operated or to LPS-treated (40 microg/kg/h for 4 h) groups. During the experiment, respiratory and circulatory parameters have been recorded and blood samples collected. At the end of the experiment the animals were sacrificed and tissue samples collected for real-time quantitative PCR and ELISA test. LPS infusion evokes a large increase in ET-1 plasma concentration, and in tissues mRNA levels, associated with an increase in pulmonary arterial pressure, as well as in pulmonary and systemic vascular resistances, and a decrease in stroke volume. LPS infusion caused also a derangement of respiratory mechanics, evidenced by an increase in resistance and a decrease in compliance of the respiratory system. ET(A) and ET(B) receptor mRNA levels were markedly decreased in liver and lung and slightly increased in heart, evidencing that ET receptor subtypes were differentially regulated in the major organs of endotoxin treated pigs. In conclusion our data show the presence of a continuative and differentially regulated stimulating mechanism of ET-1 expression during pig endotoxaemia as well as a fundamental role of ET-1 system in the cardiovascular and respiratory derangement. PMID- 16043244 TI - Seeing spots: complex phase behavior in simple membranes. AB - Liquid domains in model lipid bilayers are frequently studied as models of raft domains in cell plasma membranes. Micron-scale liquid domains are easily produced in vesicles composed of ternary mixtures of a high melting temperature lipid, a low melting temperature lipid, and cholesterol. Here, we describe the rich phase behavior observed in binary and ternary systems. We then discuss experimental challenges inherent in mapping phase diagrams of even simple lipid systems. For example, miscibility behavior varies with lipid type, lipid ratio, lipid oxidation, and level of impurity. Liquid domains are often circular, but can become noncircular when membranes are near critical points. Finally, we reflect on applications of phase diagrams in model systems to rafts in cell membranes. PMID- 16043245 TI - Excess weight at time of presentation of myocardial infarction is associated with lower initial mortality risks but higher long-term risks including recurrent re infarction and cardiac death. AB - OBJECTIVES: To evaluate the influence of elevated body mass index (BMI) on short- and long-term survival following acute myocardial infarction (AMI). BACKGROUND: Recent studies suggest an obesity survival paradox in individuals undergoing percutaneous coronary intervention with better 30-day and 1-year outcomes in obese relative to normal weight patients. We tested a similar obesity paradox hypothesis following acute myocardial infarction. METHODS: Short- and long-term all-cause mortality, and risk of recurrent AMI were evaluated according to BMI status in 894 consecutive survivors of AMI <80 years of age admitted to the Mayo Clinic Coronary Care Unit between January 1, 1988 and April 16, 2001. Normal weight, overweight and obesity were defined as BMI <25, 25-29.9, and >30 kg/m(2), respectively. RESULTS: Overall mortality following hospital discharge was significantly lower in overweight and obese patients and was mostly attributable to lower 6-month mortality (adjusted HR = 0.47, P = 0.01 for BMI >25 kg/m(2)) relative to normal weight patients, while long-term mortality among 6-month survivors was similar in all 3 groups. The risk of recurrent AMI was higher in patients with BMI >25 kg/m(2) (adjusted HR = 2.30, P = 0.01). Overweight and obese patients were significantly more likely to die from cardiac rather than non cardiac causes (P < 0.01). CONCLUSIONS: Following AMI, overweight and obese individuals although paradoxically protected from short-term death have a long term mortality risk that is similar to normal weight individuals. Younger age at the time of initial infarction and fewer non-cardiovascular comorbidities presumably explain the short-lived obesity survival paradox following myocardial infarction. PMID- 16043246 TI - Comparison of different labeling methods for the production of labeled target DNA for microarray hybridization. AB - Different labeling methods were studied to compare various approaches to the preparation of labeled target DNA for microarray experiments. The methods under investigation included a post-PCR labeling method using the Klenow fragment and a DecaLabel DNA labeling kit, the use of a Cy3-labeled forward primer in the PCR, generating either double-stranded or single-stranded PCR products, and the incorporation of Cy3-labeled dCTPs in the PCR. A microarray that had already been designed and used for the detection of microorganisms in compost was used in the study. PCR products from the organisms Burkholderia cepacia and Staphylococcus aureus were used in the comparison study, and the signals from the probes for these organisms analyzed. The highest signals were obtained when using the post PCR labeling method, although with this method, more non-specific hybridizations were found. Single-stranded PCR products that had been labeled by the incorporation of a Cy3-labeled forward primer in the PCR were found to give the next highest signals upon hybridization for a majority of the tested probes, with less non-specific hybridizations. Hybridization with double-stranded PCR product labeled with a Cy3-labeled forward primer, or labeled by the incorporation of Cy3 labeled dCTPs resulted in acceptable signal to noise ratios for all probes except the UNIV 1389a and Burkholderia genus probes, both located toward the 3' end of the 16S rRNA gene. The comparison of the different DNA labeling methods revealed that labeling via the Cy3-forward primer approach is the most appropriate of the studied methods for the preparation of labeled target DNA for our purposes. PMID- 16043247 TI - Developing implantable optical biosensors. AB - Nanobiotechnologists are developing devices that can measure specific enzymes and proteins. These devices are expected to detect single enzyme or protein molecules accurately, providing highly sensitive biosensing applications. A recent study by Strano and co-workers shows that single-walled carbon nanotubes (SWNTs) hold great promise as implantable biosensors. Although most researchers have focused on substrate-oriented biosensors, Strano and colleagues have shown that the inherent fluorescent properties of suspended individual SWNTs can be used for solution-phase beta-D-glucose sensing. PMID- 16043248 TI - Development of upper limb proprioceptive accuracy in children and adolescents. AB - This study was designed to determine differences in the proprioceptively guided movements of children (8-10 years) and adolescents (16-18 years). Participants were blindfolded and asked to actively match passively determined target positions of the elbow joint under three matching conditions. Overall, children were less accurate than adolescents in all matching tasks and utilized different kinematic strategies for making the matching movements. Specifically, children made larger absolute errors and utilized matching movements which, compared to adolescents, were of shorter duration and less irregular in terms of their velocity profiles. An assessment of limb asymmetry was also performed revealing a non-dominant arm matching advantage but only for children and only in the task requiring interhemispheric transfer of a memory-based model of limb position. The proprioceptive differences observed in this study are likely the result of experience-driven refinement in the utilization of somatosensory feedback throughout childhood and into adolescence. PMID- 16043249 TI - Use of hydrogen peroxide in combination with nisin, sodium lactate and citric acid for reducing transfer of bacterial pathogens from whole melon surfaces to fresh-cut pieces. AB - Hydrogen peroxide (2.5%) alone or hydrogen peroxide (1%) in combination with nisin (25 microg/ml), sodium lactate (1%), and citric acid (0.5%) (HPLNC) were investigated as potential sanitizers for reducing Escherichia coli O157:H7 or Listeria monocytogenes populations on whole cantaloupe and honeydew melons. Whole cantaloupes inoculated with E. coli O157:H7 and L. monocytogenes at 5.27 and 4.07 log10 CFU/cm2, respectively, and whole honeydew melons inoculated with E. coli O157:H7 and L. monocytogenes at 3.45 and 3.05 log10 CFU/cm2, respectively, were stored at 5 degrees C for 7 days. Antimicrobial washing treatments were applied to inoculated whole melons on days 0 or 7 of storage and surviving bacterial populations and the numbers transferred to fresh-cut pieces were determined. At days 0 and 7 treatment with HPLNC significantly (p<0.05) reduced the numbers of both pathogens, by 3 to 4 log CFU/cm2 on both types of whole melon. Treatment with HPLNC was significantly (p<0.05) more effective than treatment with 2.5% hydrogen peroxide. While fresh-cut pieces prepared from stored whole melons were negative for the pathogens by both direct plating and by enrichment, fresh-cut pieces from cantaloupe melons treated with 2.5% hydrogen peroxide were positive for both pathogens and pieces from honeydew melons were positive for E. coli 0157:H7. The native microflora on fresh-cut melons were also substantially reduced by HPLNC treatment of whole melons. The results suggest that HPLNC could be used to decontaminate whole melon surfaces and so improve the microbial safety and quality of fresh-cut melons. PMID- 16043250 TI - Use of flow-cytometric analysis to optimize cell banking strategies for production of biopharmaceuticals from mammalian cells. AB - Production of biopharmaceuticals from mammalian cells requires generation of master, working and post-production cell banks of high quality under GMP conditions. An optimal cryopreservation strategy is needed for each new production cell line, particularly with regard to establishing production processes that are completely devoid of serum or even any animal components and to ensuring robust thaw performance for reliable production. Here, we describe a novel strategy employing flow-cytometric (FC) analysis of Annexin V-stained cells for high-throughput characterization of cell banks. Our data show that this method enables predictive evaluation of a cryopreservation strategy as early as 6h after thawing of cells. Furthermore, a broad study is presented characterizing various factors that may influence the quality of serum-free production cell banks from NSO and CHO cell lines. These results demonstrate how FC-based analysis can be used for development of future state-of-the-art cryopreservation strategies. PMID- 16043251 TI - Isolation of promoter for N-methyltransferase gene associated with caffeine biosynthesis in Coffea canephora. AB - N-Methyltransferases (NMTs) catalyze the three SAM dependent sequential methylation of xanthosine, producing caffeine in Coffea species. In the present work, a PCR based genome walking method was adopted to isolate and clone the promoter for the NMT gene. Inspection of the promoter sequence revealed the presence of several motifs important for the regulation of the gene expression. The whole fragment was fused to the beta-glucuronidase (gus) reporter gene and used in Agrobacterium tumefaciens mediated transformation of Nicotiana tabacum. GUS assays proved that the isolated promoter was able to direct the expression of the reporter gene in transgenic tobacco. Based on the promoter sequence, primer was designed and the genomic fragment comprising the promoter and its corresponding gene was amplified and cloned. Sequencing of one of the genomic clones revealed the presence of four exons and three introns in NMT gene. The differences in the restriction pattern among the genomic clones were studied using PCR-RFLP. This is the first report of cloning of the promoter for a gene involved in caffeine biosynthetic pathway and it opens up the possibility of studying the molecular mechanisms that regulate the production of caffeine. PMID- 16043252 TI - A platform for high-throughput expression of recombinant human enzymes secreted by insect cells. AB - Functional genomics and proteomics have been fields of intense investigation, since the disclosure of the sequence of the human genome. To contribute to the assignment of a physiological role to the vast number of coding genes with unknown function, we have undertaken a program to clone, express, purify and determine the catalytic activity of those enzymes predicted to enter the secretory pathway, focusing our efforts on human peptidases. Our strategy to promote high-throughput expression and purification of recombinant proteins secreted by insect cells relies on the expression of the target enzymes with their native leader sequences and on the carboxyl-terminal fusion with a poly histidine tag. Growth of host cells were optimized in 24-well format to achieve highly paralleled culture conditions with production yields comparable to shake flask. The purification was performed by a robotic system in 96-well format using either magnetic beads or minicolumns. In a pilot study using reference peptidases and lipases, the high-throughput approach demonstrated to support the secretion in the insect cell medium of 85% of the sample enzymes. Of them, 66% have been proven to be catalytically active using fluorescent homogeneous assays in 384 well format compatible with the high-throughput screening criteria. The implications of these results are discussed in light of the application of this procedure to genomic-predicted peptidases. PMID- 16043253 TI - siRNA injection induces sequence-independent protection in Penaeus monodon against white spot syndrome virus. AB - White spot syndrome virus (WSSV) is a major disease in crustaceans, particularly shrimp, due to the current intensity of aquaculture practices. Novel strategies including vaccination to control this virus would be highly desirable. However, invertebrates lack a true adaptive immune response system and seem to rely on various innate immune responses. An alternative and more specific approach to counteract WSSV infections in shrimp could be by the exploitation of RNA interference. As long dsRNA molecules induce a general, sequence-independent anti viral immunity in shrimp [Robalino, J., Browdy, C.L., Prior, S., Metz, A., Parnell, P., Gross, P., Warr, G., 2004. J. Virol. 78, 10442-10448], it was investigated whether shorter 21 nt siRNAs with homology to the WSSV vp15 and vp28 genes would give a sequence-specific interference response in the shrimp Penaeus monodon. Vp28 siRNAs as well as nonspecific control gfp siRNAs were able to specifically and efficiently silence their homologous genes in a heterologous baculovirus insect cell expression system. However, in shrimps no such a specific effect was observed. Shrimp injected with vp15 or vp28 siRNAs before WSSV challenge gave a significantly lower mortality rate, but not significantly different when shrimps were injected with gfp siRNA. Thus, large dsRNA molecules as well as siRNAs induce a sequence-independent anti-viral immunity when injected in shrimp. PMID- 16043254 TI - The chlamydial developmental cycle. AB - Intracellular parasitism by bacterial pathogens is a complex, multi-factorial process that has been exploited successfully by a wide variety of organisms. Members of the Order Chlamydiales are obligate intracellular bacteria that are transmitted as metabolically inactive particles and must differentiate, replicate, and re-differentiate within the host cell to carry out their life cycle. Understanding the developmental cycle has been greatly advanced by the availability of complete genome sequences, DNA microarrays, and advanced cell biology techniques. Measuring transcriptional changes throughout the cycle has allowed investigators to determine the nature of the temporal gene expression changes required for bacterial growth and development. PMID- 16043255 TI - A new model validation tool using kernel regression and density estimation. AB - In physiological system modelling for control or decision support, model validation is a critical element. A nonparametric approach for assessing the validity of deterministic dynamic models against empirical data is developed, based on kernel regression and kernel density estimation, yielding visual graphical assessment tools as well as numerical metrics of compatibility between the model and the data. Nonparametric regression has been suggested for assessing a parametric statistical model by constructing a confidence band for the proposed model and then checking whether the nonparametric regression curve lies within the band. However, for deterministic models, there is no confidence band that can be constructed. A reversal of roles is therefore suggested--construct a probability band for the nonparametric regression curve and check whether the proposed model lies within the band. This approach extends the utility of nonparametric regression for model assessment to deterministic models. Weighted kernel density estimation is incorporated to derive a density profile for the regression curve, creating a local graphical validation tool. In addition, the density profile is used to define and compute two numerical measures--average normalized density (AND) and relative average normalized density (RAND), representing global statistical validity measures. These tools are demonstrated using a biomedical system model for agitation-sedation and sedation management control. PMID- 16043256 TI - Computational biomechanical modelling of the lumbar spine using marching-cubes surface smoothened finite element voxel meshing. AB - There is a need for the development of finite element (FE) models based on medical datasets, such as magnetic resonance imaging and computerized tomography in computation biomechanics. Direct conversion of graphic voxels to FE elements is a commonly used method for the generation of FE models. However, conventional voxel-based methods tend to produce models with jagged surfaces. This is a consequence of the inherent characteristics of voxel elements; such a model is unable to capture the geometries of anatomical structures satisfactorily. We have developed a robust technique for the automatic generation of voxel-based patient specific FE models. Our approach features a novel tetrahedronization scheme that incorporates marching-cubes surface smoothing together with a smooth-distortion factor (SDF). The models conform to the actual geometries of anatomical structures of a lumbar spine segment (L3). The resultant finite element analysis (FEA) at the surfaces is more accurate compared to the use of conventional voxel based generated FE models. In general, models produced by our method were superior compared to that obtained using the commercial software ScanFE. PMID- 16043257 TI - Severely diseased lung cancer patients narrate the importance of being included in a helping relationship. AB - Because patients with advanced lung cancer have a poor prognosis, healthcare staff should treat and support them with sensitivity without placing them under necessary strain. A common way of revealing patients' psychological needs is to rely on questionnaires where predefined potential problem areas are examined. Another and less common way of detecting their needs is to focus on the patients' concrete everyday-experiences in their contacts with health care. In this study, 23 consecutive patients with advanced non-small cell lung cancer were asked to describe their experiences in dealing with their healthcare providers. Data were analysed qualitatively by categorising the incidents according to content. It emerged that 'being connected to health care' and being 'acknowledged as a person' were by far the most prominent dimensions. Very few incidents were directly related to 'information'. The results suggest that in oncology it is important to call attention to the fact that the patient-physician relationship cannot be reduced to the communication of information. Other dimensions are worth considering. PMID- 16043258 TI - Promoter methylation of genes in bronchial lavages: a marker for early diagnosis of primary and relapsing non-small cell lung cancer? AB - A prospective screening program, including CT, autofluorescent bronchoscopy, biopsies and bronchial lavage (BL) collection, was initiated with the specific goal of identifying biomarkers for the early detection of non-small cell lung cancer. We report and discuss the results of p16, DAPK, MGMT, FHIT and APC methylation analysis in the 126 first patients: 77 at high risk of cancer and 49 followed up after primary cancer resection. Positive results were found in 49% of BLs, 53% in current smokers and 43% in former smokers. In presence of peripheral tumours, only 38% of BLs were abnormal versus 73% in presence of central tumours, 50% in presence of preneoplasic lesions and 47% in absence of lesions. FHIT methylation was an early event, observed in one-third of the BLs from patients with or without lesions as well as in tumours. APC methylation was a late event observed in 33% of tumours but rarely in BLs. p16 was methylated in 17% of BLs but in 48% of tumours; DAPK in 15% of BL and 22% of tumours. MGMT methylation was rare. Among patients followed up after cancer surgery, 14 were in remission with normalised BL, whereas three had positive BLs and relapsed with a central tumour. Thus, gene methylation in BL might help to detect central tumours but a CT is crucial for peripheral cancer detection. PMID- 16043259 TI - Acquisition of fluorescence and reflectance spectra during routine bronchoscopy examinations using the ClearVu Elite device: pilot study. AB - PURPOSE: Fluorescence flexible bronchoscopy (FFB) has proved to be very useful for detecting carcinoma in situ (CIS) and pre-cancerous lesions of the lung that are generally occult to white light reflectance bronchoscopy (WLRB). However, the increased sensitivity has caused a significant decrease of specificity, resulting in a large number of false positive signals that lead to a significant number of unnecessary biopsies. We have been planning to test a hypothesis that reflectance spectra and fluorescence spectra could be used to distinguish the true positive lesions from the false positive ones. To properly test such hypothesis, several thousand patients will need to be examined to obtain sufficient data from different lung lesions. Towards this goal, we have developed and have been testing a special system (ClearVu Elite ) that facilitates acquisition of both WLRB and FFB spectra during routine bronchoscopy examinations. In this pilot study we examined (1) if such could be used in a practical, routine clinical conditions without affecting commonly used bronchoscopy procedures; (2) if the spectral data obtained from the images are identical to those obtained with fiber optic probes; and (3) if the few malignant and early neoplastic lesions available for this pilot study show any differences from normal lung tissue. METHODS: The study population consisted of 63 patients that were suspicious for presence of lung cancer (19 female and 44 male, smokers and non-smokers). All were examined by diagnostic bronchoscopy using ClearVu Elite during a period of 8 weeks. RESULTS AND CONCLUSIONS: The experimental system provides excellent real time WLRB and FFB that can be switched from one to another mode instantaneously. Acquisition of both white light reflectance spectra and fluorescence spectra is acquired in real time through the images and does not prolong the routine WLRB and FFB procedures. The experimental bronchoscopy procedure is as simple as conventional bronchoscopy, adding on average less than 5 min to the 20 min procedure. The acquired spectra are identical to those obtained by fiber-optic probes. In all of the limited number of malignant and early neoplastic lesions, there were differences found which are sufficiently pronounced to warrant initiation of a large, multicenter study for development of differentiating algorithms of statistical validity. PMID- 16043260 TI - Prognostic significance of Bax and Fas ligand in erionite and asbestos induced Turkish malignant pleural mesothelioma. AB - Environmentally exposed erionite is a potent and unique inducer of malignant pleural mesothelioma (MPM) in Central Anatolia in Turkey. Previous studies have shown that erionite induced MPM has different biological behavior than asbestos induced MPM. Although impaired apoptosis has been implicated in tumor biology, the relationship between the type of environmental exposure and apoptosis has not yet been evaluated in MPM. The purpose of this study was to determine the expression of apoptosis regulating proteins and their prognostic significance in erionite and asbestos induced MPM. Thirty-five patients with MPM (16 erionite and 19 asbestos induced), and 17 patients with adenocarcinoma were comparatively evaluated. Expression of Bcl-2, Bax, Fas and Fas Ligand, were assessed by immunohistochemistry. Bcl-2 and Fas did not stain in almost all specimens. The staining extension of Bax was 13.75 +/- 19.27%, 5.89 +/- 14.51% and 7.38 +/- 14.53% for erionite and asbestos induced MPM and adenocarcinoma, respectively (p = 0.566). The staining extension of Fas Ligand was 26.87 +/- 31.87%, 46.10 +/- 37.30% and 26.47 +/- 23.23% for erionite and asbestos induced MPM, and adenocarcinoma, respectively (p = 0.123). Bax negative patients in erionite group had longer survival than Bax positive patients (18 months versus 14 months) (p = 0.06). Fas Ligand positive patients showed statistically better survival than Fas Ligand negative patients in all MPM group (15 months versus 12 months) (p = 0.05). Although all proteins expressed in similar extension in all samples, Bax staining displayed an inverse relation with survival in erionite group. This may implicate a difference in Bax functioning in erionite induced MPM. However, Fas Ligand may be functionally intact to reduce tumor survival. PMID- 16043261 TI - Oxytocin receptor pattern of expression in primary lung cancer and in normal human lung. AB - In order to assess if oxytocin- and vasopressin-induced mitogenic effects detected on small-cell lung carcinoma (SCLC) cell lines could be transposed on primary SCLC, the aim of the present work was to identify mediators of these mitogenic actions on primary tumours samples. This was addressed on normal human lung tissue, on SCLC and on non-SCLC (NSCLC). Herein, we observe, in normal human lung, that OTR is colocalized with vascular endothelial cells of the lung and is not expressed by lung cells of epithelial nature. We detected mRNA amplification of V1aR, V2R and of a V2R variant. We observed that 86% of SCLC biopsies analyzed expressed at least the OTR and that 71% expressed the OTR, the V1aR and the V2R altogether. Comparatively, 50% of NSCLC biopsies tested expressed at least the OTR and 32% expressed the OTR, the V1aR and the V2R altogether. The occurrence of the V1bR/V3R is of 28 and 18% for SCLC and NSCLC, respectively. Nevertheless, for the SCLC biopsies analyzed in this study, V1bR/V3R expression correlates, in all cases, with the expression of all the other neurohypophysial peptide receptors. Our results suggest that neurohypophysial peptide antagonists may offer promise as a potential new therapeutic modality for the treatment of lung cancer expressing at least one of the neurhypophysial peptide receptor subtypes. PMID- 16043262 TI - Long term follow up and analysis of long term survivors in patients treated with paclitaxel-based concurrent chemo/radiation therapy for locally advanced non small cell lung cancer. AB - PURPOSE: For patients with locally advanced non-small cell lung cancer (LANSCLC), concurrent chemotherapy/radiation therapy (RT) has become the standard of care. Three multi-institutional phase II studies with paclitaxel-based chemotherapeutic regimen given concurrently with RT for patients with LANSCLC were performed from March of 1994 to May of 1997. We sought to determine mature data from this database of patients, as well as to perform analysis of a cohort of patients who have achieved long term survival (LTS) when treated with this regimen. PATIENTS AND METHODS: Database of these patients was analyzed retrospectively upon longer follow up, with median follow up for the three studies being 498 days (range 11 2905 days, average 780 days). Weight loss limitation for the three studies was liberal: weight loss <10% and <15% 3 months preceding diagnosis (LUN-27 and LUN 63, respectively), and no weight loss limitation for LUN-56. RESULTS: The 4-year overall survival (OS) for the three trials was 16.3%, and 2-year progression free survival (PFS) was 25.7%. Statistical analysis of the long term survivors (OS > 4 years) was performed, and performance status (PS) was found to be a significant factor predictive of LTS. PS of 0 compared to 1 yielded a 2.5-fold increased likelihood of LTS (p = .04). There was also a trend (p = .067) for responders (complete or partial response) to yield a five-fold likelihood of LTS compared to non-responders (stable or progressive disease). CONCLUSION: Our results support the efficacy of combined modality therapy (CMT) for patients with LANSCLC even despite our more liberal weight loss eligibility criteria. Furthermore, our analysis indicates that LTS is more likely to be achievable in patients with PS = 0 compared to 1 when treated with CMT for LANSCLC. PMID- 16043263 TI - Identification of mature nocistatin and nociceptin in human brain and cerebrospinal fluid by mass spectrometry combined with affinity chromatography and HPLC. AB - Nocistatin (NST) and nociceptin/orphanin FQ (NCP) are two important bio-peptides derived from the precursor protein prepronociceptin (ppNCP), involved in several central nervous system (CNS) functions including pain transmission. Since the actual form of human NST in CNS is not fully characterized, we studied the structure of NST from human brain tissue and cerebrospinal fluid (CSF) samples. NST and NCP were isolated from human brain and CSF samples by affinity chromatography combined with HPLC. Mass spectrometry was used for the identification and characterization of the peptides. The total NST immunoreactivity was detected as 11.5+/-2.3 pmol/g tissue for the brain and 0.44 pmol/ml for the pooled CSF sample after the HPLC purification by radioimmunoassay. The presence of two different forms of mature nocistatin (NST 17 and NST-30) and a possible N-terminal methionine cleaved NST-29 were confirmed by both radioimmunoassay and mass spectrometry. Affinity chromatography, HPLC and mass spectrometry methods used in this study were highly sensitive and suitable for identification of actual chemical structures and quantification of very small amounts of peptides in biological samples. The present findings may help further for search for new treatment of neuropathic pain, which is often poorly managed by current therapies. PMID- 16043264 TI - Immunohistochemical localization of orexin-B, orexin-1 receptor, ghrelin, GHS-R in the lacrimal gland of normal and diabetic rats. AB - Orexin-B, ghrelin and their receptors play an important role in the regulation of feeding in mammals. The pattern of distribution of orexin-B, orexin-1-receptor (OX1R), ghrelin and growth hormone secretagogue receptor (GHS-R) in the lacrimal gland of normal and diabetic rats has not been reported. Diabetes was induced by a single intraperitoneal injection of streptozotocin (STZ) at 60 mg kg(-1). Forty weeks after the induction of STZ-induced diabetes, normal, age-matched controls and diabetic rats were anesthetized with chloral hydrate (intraperitoneally) and their lacrimal glands removed and processed for immunofluorescence. Orexin-B was observed in the cells localized to the interacinar regions while OX1R was discerned in the nerves innervating the wall of small blood vessels. Ghrelin was also present in a group of cells located in the periacinar regions of the lacrimal glands of normal and diabetic rats. In contrast, GHS-R was observed in the apical region of the ductal cells of the lacrimal glands of both normal and diabetic rats. The pattern of distribution of these orexigenic peptides and their receptors did not significantly change after the onset of diabetes. In conclusion, orexin-B, ghrelin and their receptors are present in the lacrimal glands of both normal and diabetic rats and may play a role in the regulation of lacrimal gland function. PMID- 16043265 TI - Angiotensin II-induced venoconstriction involves both AT1 and AT2 receptors and is counterbalanced by nitric oxide. AB - The venoconstrictor effect of Angiotensin II (Ang II) was investigated in the rat mesenteric venules and portal vein. Mesenteric venules were perfused at a constant rate and reactivity to Ang II (0.1 nmol) was evaluated as changes in the perfusion pressure. Rings of portal vein were mounted in organ baths and curves to Ang II (0.1-100 nmol/L) were generated. In venules, Ang II-contraction (10.6+/ 1.1 mmHg) was abolished by losartan (0.9+/-0.3 mmHg*), reduced by PD 123,319 (5.8+/-0.9 mmHg*), increased by L-NAME (16.5+/-1.8 mmHg*) and not altered by indomethacin. In portal veins, curves to Ang II (-logEC50: 8.9+/-0.1 mol/L) were shifted to the right by losartan (-log EC50: 7.5+/-0.1 mol/L*) and by PD 123,319 (-logEC50: 8.0+/-0.1 mol/L*). L-NAME increased the maximal response to Ang II (Emax: 0.91+/-0.1g versus 1.62+/-0.3g*) and indomethacin had no effect. In conclusion, Ang II induces venoconstriction by activating AT1 and AT2 receptors. Data obtained with L-NAME provide evidence that the basal nitric oxide release from the endothelium of the venous system can modulate the Ang II-induced venoconstriction. PMID- 16043266 TI - [Pleural effusion of vascular origin]. PMID- 16043267 TI - [Idiosyncratic drug-induced agranulocytosis]. AB - BACKGROUND: Agranulocytosis is a life-threatening disorder that frequently occurs as an adverse reaction to drugs. CURRENT DATA: Idiosyncratic drug-induced agranulocytosis is characterized by a neutrophil count <0.5x10(9)/l, in serious forms <0,1x10(9)/l that currently occurs especially in association with antibiotics, antithyroid drugs ant ticlopidine (>60% of the incriminated drugs). The overall incidence of idiosyncratic agranulocytosis ranges from 2.4 to 15.4 cases per million patients exposed to drugs per year. Although patients experiencing idiosyncratic agranulocytosis may be asymptomatic (50%), the severity of the neutropenia usually leads to severe sepsis: fever of unknown origin, septicemia, septic shock or localized documented infections such as sore throat, various cutaneous infections or pneumonia. Nevertheless, the mortality rate of idiosyncratic agranulocytosis is now around 5% with appropriate management. PERSPECTIVES: In the future, management of drug-induced agranulocytosis may include pre-established procedures using in critically situations, broad-spectrum antibiotic therapy and hematopoietic growth factors (G CSF). PMID- 16043268 TI - Guidelines to the United Kingdom Disability Discrimination Act (DDA) 1995 and the Special Educational Needs and Disability Act (SENDA) 2001 with regard to nurse education and dyslexia. AB - This paper concerns the impact of disability legislation on nurse education, nurse educators and student nurses, in relation to academic work and clinical placement, with regard to dyslexia. The two United Kingdom acts considered are the Disability Discrimination Act (DDA), 1995 and the Special Educational Needs and Disability Act (SENDA), 2001, which is an amendment to the DDA. The paper examines and defines the main points of the acts, such as discrimination; less favourable treatment and its justification; reasonable adjustments; making adjustments in advance; disclosure and confidentiality requests; substantial disadvantage; current systems and regulations and concludes by raising issues which require clarification. PMID- 16043269 TI - Modified vaccinia Ankara expressing HIVA antigen stimulates HIV-1-specific CD8 T cells in ELISpot assays of HIV-1 exposed infants. AB - Recombinant modified vaccinia virus Ankara expressing HIV-1 antigens (MVA.HIVA) was used in ELISpot assays to monitor HIV-1-specific T cell responses in infants. Responses to MVA.HIVA and HIV-1 peptides were examined in 13 infected and 81 exposed uninfected infants in Nairobi, Kenya. Responses to MVA.HIVA (38%) and peptide stimulation (38%) were similar in frequency (p=1.0) and magnitude (mean 176 versus 385 HIVSFU/10(6), p=0.96) in HIV-1 infected infants. In exposed uninfected infants, MVA.HIVA detected more positive responses and higher magnitude responses as compared to peptide. MVA.HIVA ELISpot is a sensitive method for quantification of HIV-1-specific CD8+ T cell responses in HIV-1 exposed infants. These results demonstrate the relevance of HIV-1 clade A consensus-derived immunogen HIVA for the viruses currently circulating in Nairobi. PMID- 16043270 TI - Haemolytic activities and adjuvant effect of Astragalus membranaceus saponins (AMS) on the immune responses to ovalbumin in mice. AB - In this study, the haemolytic activities of Astragalus membranaceus saponins (AMS) and its adjuvant potentials on the cellular and humoral immune responses of ICR mice against OVA were evaluated. We determined the haemolytic activity of AMS using 0.5% rabbit red blood cell. AMS showed a slight haemolytic effect, with its haemolytic percent being 0.66% at the concentration of 500 microg/ml. Furthermore, the adjuvant potentials of AMS at three dose levels on the cellular and humoral immune responses of ICR mice against ovalbumin (OVA) were investigated. ICR mice were immunized subcutaneously with OVA 100 microg alone or with OVA 100 microg dissolved in saline containing Alum (200 microg), QuilA (10 and 20 microg) or AMS (50, 100 or 200 microg) on Day 1 and 15. Two weeks later (Day 28), concanavalin A (Con A)-, lipopolysaccharide (LPS)- and OVA-stimulated splenocyte proliferation and OVA-specific antibodies in serum were measured. AMS significantly enhanced the Con A-, LPS-, and OVA-induced splenocyte proliferation in the OVA-immunized mice especially at a dose of 100 microg (P<0.05 or P<0.001). OVA-specific IgG, IgG1 and IgG2b antibody titers in serum were also significantly enhanced by AMS compared with OVA control group (P<0.01 or P<0.001). Moreover, no significant differences (P>0.05) were observed between enhancing effect of AMS and QuilA on the OVA-specific IgG, IgG1 and IgG2b antibody responses to OVA in mice. In conclusion, the results suggest that AMS could be safely used as adjuvant with low or non-haemolytic effect. PMID- 16043271 TI - Immunogenicity of an inactivated hepatitis A pediatric vaccine: three-year post booster follow-up. AB - The persistence of anti-hepatitis A virus antibody concentrations was followed over 3 years in 177 healthy children following primary and booster vaccination with an inactivated hepatitis A vaccine, Avaxim 80 pediatric. Seroconversion rates (post-immunization anti-HAV antibody concentration >or=20 mIU/mL) and geometric mean concentrations (GMC) were estimated for each of three age groups: 18 month--3 years, 4--8 years, and 9--15 years. Only subjects who were initially HAV-seronegative at inclusion (<20 mIU/mL) were analyzed. Follow-up visits at years 1, 2, and 3 involved 177, 149, and 135 children, respectively. A decline in GMCs of about 74% occurred during the first year, from 3,060 to 814 mIU/mL overall, but did not continue during years 2 and 3. All subjects remained seropositive (antibody concentration >or=20 mIU/mL), with overall GMCs of 814, 891, and 924 mIU/mL in years 1--3, respectively. The inactivated hepatitis A study-vaccine resulted in sustained seroprotective antibody concentrations in 100% of these children, without a significant decline in antibiotic concentrations over the 3 years following booster injection, thus demonstrating the long-term protection expected with this vaccine. PMID- 16043272 TI - Zinc and copper uptake by plants under two transpiration rates. Part II. Buckwheat (Fagopyrum esculentum L.). AB - To evaluate the environmental risks of irrigating crops with treated wastewater, a study was undertaken to quantify heavy metal uptake by 4-week old buckwheat (Fagopyrum esculentum L.) plants during 18 days of irrigation with 8 different Cu and Zn solutions under two transpiration rates (TR). At 4 weeks, potted buckwheat plants were transferred into one of the two growth chambers, offering either a high or low vapour pressure deficit (VDP) for, respectively, a high or low TR. Triplicate pots received one of the 8 irrigation treatments containing one of two Zn levels (0 and 25 mg/L) combined with one of four Cu levels (0, 5, 10 and 15 mg/L). Daily TR were measured by weighing the evapo-transpired water lost from the planted pot, less was the evaporation loss measured from triplicate non planted pots. After 0, 6, 12 and 18 days of treatment, the stems and leaves of three randomly selected plants were harvested and after 18 days, the roots were harvested to determine Cu and Zn uptake. The treatments did not affect TR in terms of dry plant mass, indicating the absence of toxic effects. Irrigating with Zn, without Cu, increased dry biomass production, whereas the lowest biomass occurred with 15 and 30 mg/L of Cu with and without 25 mg/L of Zn, respectively, because higher applications of heavy metal significantly reduced soil pH. Plant Cu and Zn uptake increased with TR. With higher levels of Cu, Zn uptake by buckwheat was significantly reduced, while Zn had a slight but non-significant impact on Cu uptake. Previously and in a study exposing wheat plants to the same conditions, Cu significantly increased Zn uptake, while Zn had a slight but insignificant negative effect on Cu uptake. The buckwheat roots contained the greatest levels of Cu and Zn, indicating their role in moderating heavy metal uptake. Also, both Cu and Zn had a synergetic effect on each other in terms of root levels, and a similar observation was made in the earlier similar experiment using wheat plants. Irrigating a buckwheat crop with treated wastewater, with more natural Cu and Zn levels of 0.08 mg/L, could be quite beneficial without endangering the quality of the crop and acidifying the soil pH. The most concentrated experimental solutions contained 300 times more Cu and Zn, to obtain measurable differences. PMID- 16043273 TI - Zinc and copper uptake by plants under two transpiration rates. Part I. Wheat (Triticum aestivum L.). AB - To evaluate the environmental risk of irrigating crops with treated wastewater, an experiment was conducted using two growth chambers, each offering a different vapour pressure deficit (VPD) for high and low transpiration rates (TR), respectively. One of the two sets of 24 pots planted with 6 week old wheat (Triticum aestivum L.), was placed in each growth chamber, and irrigated in triplicates for 20 days with 8 Zn and Cu solutions (0 and 25 mg Zn/L combined with 0, 5, 15 and 30 mg Cu/L). Water losses from planted and non-planted pots served to measure evapo-transpiration and evaporation, respectively. Pots were monitored for Cu and Zn uptake by collecting three plants (shoot and grain)/pots after 0, 10 and 20 days, and roots in each pot after 20 days, and analyzing these plant parts for dry mass, and Cu and Zn levels. Transpiration rate was not affected by any Cu/Zn treatment, but Cu and Zn uptake increase with the time, irrigation solution level and higher TR, with the roots retaining most Cu and Zn, compared to the shoot followed by the grain. For the shoot and grain, Cu had a significant synergetic effect on Zn uptake, when Zn had slight but insignificant antagonistic effects on Cu uptake. For the roots, Cu and Zn had significant synergetic effect on each other. Regression equations obtained from the data indicate that Cu and Zn levels normally found in treated wastewater (0.08 mg/L) are 300 times lower than those used for the most concentrated experimental solutions (30 and 25 mg/L, respectively) and may, on a long term basis, be beneficial rather than toxic to wheat plants and do not acidify soil pH. PMID- 16043274 TI - Spatial variation in the degradation rate of the pesticides isoproturon, azoxystrobin and diflufenican in soil and its relationship with chemical and microbial properties. AB - The extent of within field variability in the degradation rate of the pesticides isoproturon, azoxystrobin and diflufenican, and the role of intrinsic soil factors and technical errors in contributing to the variability, was investigated in sites on sandy-loam and clay-loam. At each site, 40 topsoil samples were taken from a 160 x 60 m area, and pesticides applied in the laboratory. Time to 25% dissipation (DT25) ranged between 13 and 61 weeks for diflufenican, 5.6 and 17.2 weeks for azoxystrobin, and 0.3 and 12.5 weeks for isoproturon. Variability in DT25 was higher in the sandy-loam in which there was also greatest variability in soil chemical and microbial properties. Technical error associated with pesticide extraction, analysis and lack of model fit during derivation of DT25 accounted for between 5.3 and 25.8% of the variability for isoproturon and azoxystrobin, but could account for almost all the variability for diflufenican. Azoxystrobin DT25, sorption and pH were significantly correlated. PMID- 16043275 TI - Weighing in primary-care nurse-patient interactions. AB - This article analyzes the interactions through which primary-care nurses and patients accomplish patient weighing. The analysis is based on videotaped nurse adult patient interactions in clinics in the area of Southern California. Detailed examination of co-participants' naturally situated weighing conduct shows that parties recurrently deliver utterances that go beyond that required to accomplish weight measurement-precisely "where" they "are" within the weighing process shaping how they produce and understand these utterances. Using weighing as a locus of epistemic negotiation and potential affiliation, co-participants interactionally achieve the distribution of weight/weighing knowledge and the character of their social relationship. Confronting their numerical weight results in a social/medical setting, patients can use expansive weighing utterances to claim or demonstrate that they possess pre-existing knowledge regarding weight, asserting independent expertise vis-a-vis nurses and claiming result co-recipiency and co-ownership. Speakers can also use expansive utterances to proffer an interactional opportunity for affiliation, inviting recipients to collaborate in producing a more personalized encounter. Through the acceptance or declination of these invitations, the parties work out "who" they "are" to and for one another. PMID- 16043276 TI - Sensory temporal processing in adults with early hearing loss. AB - This study examined tactile and visual temporal processing in adults with early loss of hearing. The tactile task consisted of punctate stimulations that were delivered to one or both hands by a mechanical tactile stimulator. Pairs of light emitting diodes were presented on a display for visual stimulation. Responses consisted of YES or NO judgments as to whether the onset of the pairs of stimuli was perceived simultaneously or non-simultaneously. Tactile and visual temporal thresholds were significantly higher for the deaf group when compared to controls. In contrast to controls, tactile and visual temporal thresholds for the deaf group did not differ when presentation locations were examined. Overall findings of this study support the notion that temporal processing is compromised following early deafness regardless of the spatial location in which the stimuli are presented. PMID- 16043277 TI - Evidence for unusual spatial location coding in Williams syndrome: an explanation for the local bias in visuo-spatial construction tasks? AB - Individuals with Williams syndrome (WS) display poor visuo-spatial cognition relative to verbal abilities. Furthermore, whilst perceptual abilities are delayed, visuo-spatial construction abilities are comparatively even weaker, and are characterised by a local bias. We investigated whether this differentiation in visuo-spatial abilities can be explained by a deficit in coding spatial location in WS. This can be measured by assessing participants' understanding of the spatial relations between objects within a visual scene. Coordinate and categorical spatial relations were investigated independently in four participant groups: 21 individuals with WS; 21 typically developing (TD) children matched for non-verbal ability; 20 typically developing controls of a lower non-verbal ability; and 21 adults. A third task measured understanding of visual colour relations. Results indicated first, that the comprehension of categorical and coordinate spatial relations is equally poor in WS. Second, that the comprehension of visual relations is also at an equivalent level to spatial relational understanding in this population. These results can explain the difference in performance on visuo-spatial perception and construction tasks in WS. In addition, both the WS and control groups displayed response biases in the spatial tasks. However, the direction of bias differed across the groups. This finding is explored in relation to current theories of spatial location coding. PMID- 16043278 TI - Central control of penile erection: role of the paraventricular nucleus of the hypothalamus. AB - The paraventricular nucleus of the hypothalamus is an integration centre between the central and peripheral autonomic nervous systems. It is involved in numerous functions from feeding, metabolic balance, blood pressure and heart rate, to erectile function and sexual behaviour. In particular, a group of oxytocinergic neurons originating in this nucleus and projecting to extra-hypothalamic brain areas (e.g., hippocampus, medulla oblongata and spinal cord) control penile erection in male rats. Activation of these neurons by dopamine and its agonists, excitatory amino acids (N-methyl-D-aspartic acid) or oxytocin itself, or by electrical stimulation leads to penile erection, while their inhibition by gamma amino-butyric acid (GABA) and its agonists or by opioid peptides and opiate-like drugs inhibits this sexual response. The activation of these neurons is secondary to the activation of nitric oxide synthase, which produces nitric oxide. Nitric oxide in turn causes, by a mechanism that is as yet unidentified, the release of oxytocin in extra-hypothalamic brain areas. Other compounds recently identified that facilitate penile erection by activating central oxytocinergic neurons are peptide analogues of hexarelin, a growth hormone releasing peptide, pro-VGF derived peptides, endogenous peptides that may be released by neuronal nerve endings impinging on oxytocinergic cell bodies, SR 141716A, a cannabinoid CB1 receptor antagonist, and, less convincingly, adrenocorticotropin-melanocyte stimulating hormone (ACTH-MSH)-related peptides. Paraventricular oxytocinergic neurons and similar mechanisms are also involved in penile erection occurring in physiological contexts, namely noncontact erections that occur in male rats in the presence of an inaccessible receptive female, and during copulation. These findings show that the paraventricular nucleus of the hypothalamus plays an important role in the control of erectile function and sexual activity. As the male rat is a model of sexual behaviour and penile physiology, which has largely increased in the last years our knowledge of peripheral and central mechanisms controlling erectile function (drugs that induce penile erection in male rats usually do so also in man), the above results may have great significance in terms of a human perspective for the treatment of erectile dysfunction. PMID- 16043279 TI - Electrophysiological correlates of categorization: P300 amplitude as index of target similarity. AB - Two experiments examined event-related potentials (ERPs) and behavioral correlates of categorizing stimuli varying in perceptual similarity to targets. Participants performed a target-detection task in which non-target stimuli varied in target similarity but occurred with equivalent probability. The stimuli were variations of a schematic human face comprised of eight distinct features: two eyes, two eyebrows, one nose, one mouth, and two ears. Non-target stimuli that were perceptually similar to targets produced larger P300-like neurophysiological responses than did other non-target stimuli. These effects emerged whether participants' target was relatively complex (eight features) or quite simple (zero features). Accordingly, the presence of many constituent elements of a test stimulus does not appear necessary to trigger increases in categorical processing of non-targets that are similar to a target. The data further suggest that the P300 amplitude may be used as a good index of perceptual similarity between target and non-target stimuli. PMID- 16043280 TI - The role of molecular chaperones in mouse sperm-egg interactions. AB - Fertilization is a unique and exquisitely choreographed cellular interaction between the male and female gamete that results in the creation of a genetically unique individual. Despite the fundamental importance of fertilization, there remains a dearth of information about the basic biochemical mechanisms that underpin this process. One of the key issues that remain unresolved is the molecular basis of sperm-egg recognition. From the female perspective, it is well established that the sperm recognition sites reside in the zona pellucida (ZP), an acellular coat that surrounds the oocyte. In contrast, numerous studies into the cognate zona receptors residing on the sperm surface have failed to shed significant light on the biochemical identity of these molecules. Such difficulties may, in part, have arisen because investigations have traditionally been based on the precept that the zona receptor represents a single molecular entity that is constitutively expressed on the sperm surface. While such a view holds obvious appeal, it fails to account for growing evidence that gamete interaction is not mediated by a simple lock-and-key mechanism. In this review, we present a novel hypothesis in which the zona recognition site is portrayed as a multimeric molecular structure that is assembled into a functional complex during a maturation process known as 'capacitation'. Furthermore, we consider the possibility that this previously cryptic complex is assembled and delivered to the outer surface of the sperm plasma membrane through the concerted action of several members of the molecular chaperone family of proteins. PMID- 16043281 TI - Modelling the leaching of Pb, Cd, As, and Cr from cementitious waste using PHREEQC. AB - A leaching model was developed using the United States Geological Survey public domain PHREEQC geochemical package to simulate the leaching of Pb, Cd, As, and Cr from cementitious wastes. The model utilises both kinetic terms and equilibrium thermodynamics of key compounds and provides information on leachate and precipitate speciation. The model was able to predict the leaching of Pb, Cd, As, and Cr from cement in the presence of both simple (0.1 and 0.6M acetic acid) and complex municipal landfill leachates. Heavy metal complexation by the municipal landfill leachate was accounted for by the introduction of a monoprotic organic species into the model. The model indicated Pb and As were predominantly incorporated within the calcium silicate hydrate matrix while a greater portion of Cd was seen to exist as discrete particles in the cement pores and Cr (VI) existed mostly as free CrO4(2-) ions. Precipitation was found to be the dominant mechanism controlling heavy metal solubility with carbonate and silicate species governing the solubility of Pb and carbonate, silicate and hydroxide species governing the solubility of Cd. In the presence of acetic acid, at low pH values Pb and Cd acetate complexes were predominant whereas, at high pH values, hydroxide species dominated. At high pH values, the concentration of As in the leachate was governed by the solubility of Ca3(AsO4)2 with the presence of carbonate alkalinity competing with arsenate for Ca ions. In the presence of municipal landfill leachate, Pb and Cd organic complexes dominated the heavy metal species in solution. The reduction of As and Cr in municipal landfill leachate was crucial for determining aqueous speciation, with typical municipal landfill conditions providing the reduced forms of As and Cr. PMID- 16043282 TI - Effects of sleep deprivation on event-related fields and alpha activity during rhythmic force production. AB - The influence of sleep deprivation (SD) on event-related fields and the distribution of power over the scalp of MEG imaged brain activity was studied during acoustically paced rhythmic force production. At the behavioral level, SD resulted in a reduction of the lag (negative asynchrony) between produced forces and acoustic stimuli at higher movement tempos. Principal component analysis of the accompanying MEG activity showed that auditory- and motor-evoked fields were attenuated after SD and revealed an anterior shift of power towards more frontal channels. These results were interpreted in terms of a change of central processing of afferent sensory input due to SD. PMID- 16043283 TI - COMT Val158Met polymorphism in schizophrenia with obsessive-compulsive disorder: a case-control study. AB - This is the first study of a possible molecular genetic basis for schizophrenia with obsessive-compulsive disorder (OCD). We performed a case-control association study of the catechol-O-methyltransferase (COMT) Val158Met polymorphism in schizophrenia-OCD patients, OCD and healthy controls. One hundred and thirteen schizophrenia-OCD patients, 79 OCD patients and 171 control subjects were genotyped for the Val(158)Met polymorphism in the COMT gene. There was no significant difference in allele and genotype distribution of the COMT gene between schizophrenia-OCD patients and healthy controls. The low-activity Met allele and Met/Met genotype were more frequent in OCD men than in schizophrenia OCD and control individuals. This difference, however, was not statistically significant following correction for multiple comparisons. These results do not support the hypothesis that the COMT Val158Met gene polymorphism is associated with liability to schizophrenia-OCD. PMID- 16043284 TI - Association between acyl-coenzyme A: cholesterol acyltransferase gene and risk for Alzheimer's disease in Chinese. AB - There is a compelling body of evidence indicating an association between cholesterol and Alzheimer's disease (AD). Acyl-coenzyme A: cholesterol acyltransferase 1 (ACAT1), an endoplasmic-reticulum-resident enzyme that catalyses the formation of cholesteryl esters (CEs) from cholesterol and long chain fatty acids, modulates the generation of beta amyloid peptide (Abeta). A single nucleotide polymorphism rs1044925 in the sterol O-acyltransferase 1 (SOAT1), the gene encoding ACAT1, has been reported to be association with an increased risk for sporadic AD (SAD) in European population. In the present study, we examined the association of the SOAT1 rs1044925 polymorphism with SAD in our northern Han-Chinese (107 cases, 118 age and gender-matched controls) sample using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. There was no genotypic (chi(2)=0.030, OR 0.942, 95% CI=0.478 1.857) or allelic (chi(2)=0.021, OR 0.955, 95% CI=0.508-1.794) association between SAD and controls, even when the data were stratified by APOEvarepsilon4 carrier status. Our results indicate that the polymorphism rs1044925 in the 3'UTR of SOAT1 gene does not affect the risk of SAD in the northern Han-Chinese. PMID- 16043285 TI - Morphological evidence for GABA/glycine-cocontaining terminals in synaptic contact with neurokinin-1 receptor-expressing neurons in the sacral dorsal commissural nucleus of the rat. AB - Previous studies have shown that neurons in the sacral dorsal commissural nucleus (SDCN) express neurokinin-1 receptor (NK1R) and can be modulated by the co release of GABA and glycine (Gly) from single presynaptic terminal. These results raise the possibility that GABA/Gly-cocontaining terminals might make synaptic contacts with NK1R-expressing neurons in the SDCN. In order to provide morphological evidence for this hypothesis, the triple-immunohistochemical studies were performed in the SDCN. Triple-immunofluorescence histochemical study showed that some axon terminals in close association with NK1R-immunopositive (NK1R-ip) neurons in the SDCN were immunopositive for both glutamic acid decarboxylase (GAD) and glycine transporter 2 (GlyT2). In electron microscopic dual- and triple-immunohistochemistry for GAD/GlyT2, GAD/NK1R, GlyT2/NK1R, or GAD/GlyT2/NK1R also revealed dually labeled (GAD/GlyT2-ip) synaptic terminals upon SDCN neurons, as well as GAD- and/or GlyT2-ip axon terminals in synaptic contact with NK1R-ip SDCN neurons. These results suggested that some synaptic terminals upon NK1R-expressing SDCN neurons co-released both GABA and Gly. PMID- 16043286 TI - Alteration of striatal [11C]raclopride and 6-[18F]fluoro-L-3,4 dihydroxyphenylalanine uptake precedes development of methamphetamine-induced rotation following unilateral 6-hydroxydopamine lesions of medial forebrain bundle in rats. AB - We studied the positron emission tomography (PET) tracer distributions of ligands for dopamine D1 receptors ([11C]SCH23390) and D2 receptors ([11C]raclopride) and of the dopamine precursor analog 6-[18F]fluoro-L-3,4-dihydroxyphenylalanine ([18F]FDOPA) in the brain after 6-hydroxydopamine (6-OHDA) lesions of the medial forebrain bundle in rats. The number of methamphetamine-induced rotation was higher at 14 days than at 3 days after the 6-OHDA lesions. The brains of 6-OHDA treated rats were analyzed by tissue dissection following i.v. bolus of each tracer at 3 days (acute stage) or 3 weeks (chronic stage) postlesion. [11C]Raclopride, but not [11C]SCH23390, showed higher accumulation in the striatum on the lesion side than on the non-lesion (intact) side both at 3 days and 3 weeks postlesion. On the other hand, lower accumulation of [18F]FDOPA was observed in the striatum on the lesion side at 3 days postlesion and in both the striatum and cerebral cortex on the lesion side at 3 weeks postlesion. Our studies demonstrate that an increase in [11C]raclopride and a decrease in [18F]FDOPA uptake in the denervated striatum is evident even at 3 days after the 6-OHDA lesions when the methamphetamine-induced rotational behavior is not established. PMID- 16043287 TI - The role of Na(V)1.8 sodium channel in the maintenance of chronic inflammatory hypernociception. AB - We previously described an animal model of persistent inflammatory sensitization of nociceptors. In this model the hypernociception persists for more than 30 days after the cessation of 2 weeks of daily intraplantar treatment with prostaglandin E(2) (PGE(2)). The tetrodotoxin-resistant (TTX-R) voltage-gated sodium channel Na(V)1.8 is considered a characteristic of primary afferent nociceptive C fibers and plays an important role in acute hypernociception. In the present study, the relevance of the Na(V)1.8 channel was investigated in this model of persistent mechanical hypernociception in rats. In the PGE(2)-induced persistent hypernociception, but not in the single injection-induced acute hypernociception, the mRNA expression (RT-PCR) of Na(V)1.8 in dorsal root ganglia (DRG) was up regulated. A similar increase of Na(V)1.8 mRNA was observed when DbcAMP was used to induce persistent hypernociception. Four daily intrathecal administrations of oligodeoxynucleotides (ODN) antisense against Na(V)1.8 decreased the mRNA encoding Na(V)1.8 in DRG. The intrathecal administration of ODN antisense prevented the PGE(2)-induced acute hypernociception and significantly reduced ongoing PGE(2)-induced persistent hypernociception. A parallel restoration of the persistent hypernociception and up-regulation of Na(V)1.8 mRNA was observed after the cessation of ODN antisense treatment. These results suggest the participation of Na(V)1.8 channels in the development and maintenance of chronic inflammatory hyperalgesia, and confirm their involvement in the acute inflammatory hypernociception. PMID- 16043288 TI - Comment on: Acupuncture analgesia during surgery: a systematic review by Hyangsook Lee and Edzard Ernst, Pain 114 (2005) 511-517. PMID- 16043289 TI - Microarray analysis of gene expression following the formalin test in the infant rat. AB - Injury and pain experienced by the infant results in immediate changes in pain sensitivity that last into adulthood. These long-term changes are likely initiated by altered gene expression. Here we measured how injury alters gene expression in the lumbar spinal cord early and late in the preweaning period of the rat. The expression of large numbers of genes was altered significantly at 3 days of age, when injury has long-term consequences. The functional classes of altered genes included transcription factors, cell death related and metal ion genes. The intensity of the stimulus in the 3-day-old pups induced changes in different classes of genes. Fewer changes were noted at 21 days of age. The increased expression of transcription factors and decreased expression of genes whose products are protective against cell death are hypothesized to underlie the long-term changes that are seen after injury in the neonate. PMID- 16043290 TI - Heat and mechanical hyperalgesia in mice model of cancer pain. AB - We developed a mouse model of cancer pain to investigate its underlying mechanisms. SCC-7, squamous cell carcinoma (SCC) derived from C3H mice, was inoculated subcutaneously into either the plantar region or thigh in male C3H/Hej mice. Heat and mechanical sensitivity as well as spontaneous behavior were measured at the plantar surface of the ipsilateral hind paw after the inoculation. Inoculated sites were histologically examined, and the expression of capsaicin receptors (TRPV1) was examined in the dorsal root ganglia (DRG) to clarify their potential contribution to pain sensitivity. Inoculation of cancer cells induced marked heat hyperalgesia and mechanical allodynia in the ipsilateral hind paw for two weeks in both plantar- and thigh-inoculation models. Signs of spontaneous pain, such as lifting, licking and flinching of the paw were also observed. However, further growth of the tumor reversed the mechanical allodynia in both plantar- and thigh-inoculation models, and heat hyperalgesia in thigh-inoculation models. Histologically, no infiltration of the tumor cells into the nerve was observed. TRPV1 immunoreactive cells increased in the L5 DRG on day 7, but returned to the control level on day 15 post-inoculation. Intraperitoneal administration of the competitive TRPV1 antagonist capsazepine inhibited hyperalgesia induced by tumor cell-inoculation in either plantar- or thigh inoculated animals. This study indicated that inoculation of SCC resulted in spontaneous pain, heat hyperalgesia and mechanical allodynia. The altered expression of TRPV1 in the DRG may be involved in behavioral changes in this model. PMID- 16043291 TI - Passive coping is a risk factor for disabling neck or low back pain. AB - BACKGROUND: Despite evidence suggesting that coping is an important concept in the study of pain, its role in predicting the development of disabling pain has not been previously studied. To assess the relationship between coping and the development of disabling pain. METHODS: From a random sample of adults, we formed a cohort of individuals with non-disabling neck and/or low back pain (n=571). Participants were followed 6 and 12 months after the index survey. Coping was measured with the Vanderbilt Pain Management Inventory. The Chronic Pain Questionnaire was used to measure the presence of disabling neck and/or low back pain. We used Cox proportional hazards regression analyses to investigate the role of passive coping in the development of disabling pain while controlling for confounders. RESULTS: Passive coping was a strong, independent risk factor for disabling neck and/or back pain. Those using moderate to high levels of passive coping strategies were at an over five-fold increased risk of developing disabling pain (Moderate: HRR=5.19, 95% CI=1.78-15.1; High: HRR=6.80, 95% CI=2.36 19.6). Active coping was not found to be a significant risk factor for disabling neck and/or back pain. CONCLUSION: Passive coping is a strong and independent predictor of disabling neck and/or back pain. This strong relationship identifies passive coping as a marker for risk of disability and can allow for the identification of individuals at risk and in need of intervention to aid in improving their overall adjustment. PMID- 16043292 TI - Effect of persistent monoarthritis of the temporomandibular joint region on acute mustard oil-induced excitation of trigeminal subnucleus caudalis neurons in male and female rats. AB - The effect of persistent inflammation of the temporomandibular (TMJ) region on Fos-like immunoreactivity (Fos-LI) evoked by acute noxious stimulation of the same or opposite TMJ was assessed in male and cycling female rats. Two weeks after inflammation of the TMJ by complete Freund's adjuvant (CFA, 25 microg) the selective small fiber excitant, mustard oil (MO, 20%), was injected into the arthritic or opposite TMJ under barbiturate anesthesia. MO stimulation of the arthritic TMJ increased Fos-LI ipsilateral, but not contralateral, to MO compared to naive subjects in superficial laminae at the trigeminal subnucleus caudalis/upper cervical cord (Vc/C2) junction independent of sex hormone status. Unexpectedly, MO stimulation of the opposite TMJ in arthritic rats also produced a greater Fos-LI response ipsilateral to MO than naive animals. Fos-LI produced in the dorsal paratrigeminal region (dPa5) and Vc/C2 junction after MO stimulation of the normal TMJ was significantly greater in proestrous than diestrous females or male monoarthritic rats. In contrast to naive animals, Fos LI was produced in deep laminae at the Vc/C2 junction ipsilateral to MO in CFA treated animals independent of the site of prior CFA inflammation or sex hormone status. These results indicated that persistent monoarthritis of the TMJ region enhanced the excitability of trigeminal brainstem neurons to subsequent TMJ injury that occurred bilaterally in multiple regions of the lower trigeminal brainstem complex and depended on sex hormone status. PMID- 16043293 TI - Inhibition of nociceptive dural input in the trigeminal nucleus caudalis by somatostatin receptor blockade in the posterior hypothalamus. AB - Somatostatin is a neuromodulator in the central nervous system and is involved in the regulation of metabolic and neuroendocrine functions. Recent experimental and clinical findings point to a role for somatostatin in the central processing of nociception. We studied the effects of somatostatin receptor modulation in the posterior hypothalamic area (PH) of the rat on dural nociceptive input. Somatostatin (10 microg/microl) and the somatostatin antagonist cyclo somatostatin (50 microg/microl) were microinjected into the PH and the effects on responses of neurons in the trigeminal subnucleus caudalis studied. Injection of somatostatin (n=11) did not affect A- and C-fibre responses to dural electrical stimulation, nor was spontaneous activity altered (P>0.05). Injection of cyclo somatostatin (n=10) into the PH reduced A-(-35.5+/-5.8%) and C-fibre (-43.1+/ 7.5%) responses to dural stimulation and resulted in decreased spontaneous activity (-38.1+/-7.3%, P<0.05). Responses to facial thermal stimulation were decreased by 51.2+/-5.8% (n=5). Control injections had no significant effect (n=9). Blockade of somatostatin receptors in the PH has an anti-nociceptive effect on dural and facial input, probably mediated via GABAergic mechanisms. As somatostatin is also involved in hypothalamic regulation of metabolic, neuroendocrine and autonomic functions, somatostatin receptor mechanisms in the PH may play a role in the pathophysiology of primary headache disorders, such as migraine or cluster headache. PMID- 16043294 TI - Multiple types of sensory neurons respond to irritating volatile organic compounds (VOCs): calcium fluorimetry of trigeminal ganglion neurons. AB - Many volatile organic compounds (VOCs) are significant environmental irritants that stimulate somatosensory nerve endings to produce pain and irritation. We measured intracellular calcium in cultured trigeminal ganglion neurons to characterize the cellular mechanisms and chemical structural determinants underlying sensitivity to VOCs. Trigeminal neurons responded to homologous series of alcohols (C4-C7) as well as saturated and unsaturated aldehydes in a concentration dependent manner. Ranked in terms of threshold to recruit neurons by compounds of the same carbon chain length, enaldehyde or = 120) and low IQ (< or = 90) groups. EEG variables at P<.05 were entered into a factor analysis and then the single highest loading variable on each factor was entered into a discriminant analysis where groups were high IQ vs. low.Q. RESULTS: Discriminant analysis of high vs. low IQ was 92.81-97.14% accurate. Discriminant scores of intermediate IQ subjects (i.e. 90 < IQ < 120) were intermediate between the high and low IQ groups. Linear regression predictions of IQ significantly correlated with the discriminant scores (r = 0.818-0.825, P < 10(-6)). The ranking of effect size was EEG phase > EEG coherence > EEG amplitude asymmetry > absolute power > relative power and power ratios. The strongest correlations to IQ were short EEG phase delays in the frontal lobes and long phase delays in the posterior cortical regions, reduced coherence and increased absolute power. CONCLUSIONS: The findings are consistent with increased neural efficiency and increased brain complexity as positively related to intelligence, and with frontal lobe synchronization of neural resources as a significant contributing factor to EEG and intelligence correlations. SIGNIFICANCE: Quantitative EEG predictions of intelligence provide medium to strong effect size estimates of cognitive functioning while simultaneously revealing a deeper understanding of the neurophysiological substrates of intelligence. PMID- 16043404 TI - Evaluating the effects of spatial frequency on migraines by using pattern reversal visual evoked potentials. AB - OBJECTIVE: To clarify the effects of contrast and spatial frequency in patients with migraine by means of pattern-reversal visual evoked potentials (PVEPs). METHODS: PVEPs were obtained from 14 patients who had migraine without aura (MO), 11 patients who had migraine with aura (MA), and 25 age-matched, healthy controls (CO). PVEPs were binocularly recorded with a reversal rate of 1Hz (2 reversal/s) at 3 spatial frequencies (0.5, 1.0 and 4.0 cpd) at high (98%), medium (83%) and low (29%) contrast. N75, P100 and N135 latency and the amplitudes of P50-N75, N75 P100 and P100-N135 were analyzed. RESULTS: Increased amplitude of PVEPs in patients with migraines were revealed at 3 different spatial frequencies in all components. The MO and the MA showed increased amplitudes mostly in high contrasts (98%). These findings were detected more at a high spatial frequency (4.0 cpd) than at a low spatial frequency (0.5 cpd). Increased amplitude with prolonged latency of N135 were found both in MO and MA at 4.0 cpd. CONCLUSIONS: We conclude that pattern stimuli of high contrasts may be particularly effective in uncovering abnormal cortical reactivity which may be modified in the primary and secondary visual cortex in the interictal state of migraine. SIGNIFICANCE: These findings indicate that there is abnormal visual cortex processing in patients with migraine. PMID- 16043405 TI - Is hypertension a predictor for heart failure? A cross cultural comparison over a 10-year period. AB - This was a retrospective cohort study to assess the effect of hypertension (HTN) among Qatari and Asian patients admitted to the Hamad General Hospital in Qatar with heart failure (HF) and to identify risk factors that contribute to the development of HF in HTN patients in the State of Qatar. A total of 20,856 patients were treated during the 10-year period; 8446 were Qataris and 60% were male. Among the total Qatari patients admitted with HF (n=2342), 52.4% had HTN. The incidence of HTN was slightly higher in males than in females (50.4 vs. 49.6%; p<0.001). Significantly more HTN patients had diabetes mellitus (DM) (p<0.001) and hypercholesterolemia (p<0.001). There was also a significant difference between Qatari and Asian HTN patients in respect of their age (p<0.001) and gender (p<0.001). Qatari hypertensive patients were more likely to have DM (p<0.001). HTN and DM were the most common risk factors for HF. PMID- 16043406 TI - The influence of aetiology on inflammatory and neurohumoral activation in patients with severe heart failure: a prospective study comparing Chagas' heart disease and idiopathic dilated cardiomyopathy. AB - Patients with Chagas' cardiomyopathy have the worst prognosis when compared to other aetiologies. It has been suggested that a more intense inflammatory activation could be responsible for this excessive mortality. We studied 35 patients with idiopathic dilated cardiomyopathy (IDC group) and 28 patients with Chagas' heart disease (Chagas' group) and 12 control subjects. We compared plasma tumor necrosis factor alpha (TNF-alpha), soluble TNF-alpha receptor type 1 (sTNF R1), soluble Fas (sFas), interleukin 6 (IL-6), and brain natriuretic peptide type B (BNP) concentrations between the groups. TNF-alpha and IL-6 concentrations were higher in the IDC and Chagas groups as compared to controls (p<0.001 and p=0.001, respectively). sTNF-R1 concentration was higher in IDC after stratification for functional class (p=0.039), and there was a trend toward higher plasma TNF-alpha concentration in the Chagas' group (p=0.092). IL-6 concentration was higher in Chagas than in IDC (p=0.005). Higher IL-6 levels were associated with worse outcome (p=0.03 for Chagas; p=0.003 for IDC). sFas concentration was similar among groups. BNP concentrations were higher in IDC (350 pg/ml) and in Chagas (444.6 pg/ml) as compared to the controls (20.3 pg/ml; p<0.01). Higher BNP levels were associated with death and heart transplantation in both aetiologies. Inflammatory activation in Chagas heart disease differs from IDC and is associated with heart failure severity. PMID- 16043407 TI - Do metoprolol and carvedilol have equivalent effects on diurnal heart rate in patients with chronic heart failure? AB - BACKGROUND: Carvedilol exerted a greater reduction in mortality than metoprolol tartrate in the Carvedilol or Metoprolol European Trial (COMET). However, it is unclear if the degree and time course of beta1-blockade during a 24-h period was similar with each agent at the doses used. Therefore we analyzed 24-h ECG Holter recordings from a study which compared the long-term clinical efficacy of metoprolol tartrate to carvedilol in chronic heart failure patients using the same dosing regimen as in COMET. METHODS AND RESULTS: Fifty-one patients with chronic heart failure with a mean LVEF 26+/-1.8% were randomized in a double blind fashion to receive metoprolol tartrate 50 mg bid or carvedilol 25 mg bid. 24-h ECG monitoring (Holter) was performed at baseline, 12 weeks and 1 year. Adequate quality recordings for analysis were obtained from 43 subjects at baseline, 42 at 12 weeks and 29 subjects at 1 year. Both drugs produced a fall in average 24-h heart rate from baseline at 12 weeks and at 1 year: metoprolol 88+/ 3 to 71+/-2 and 69+/-3 bpm; carvedilol 83+/-3 to 70+/-2 and 70+/-3 bpm respectively (all p<0.001). The pattern of suppression of heart rate during the 24-h period was similar for both drugs. CONCLUSION: Metoprolol tartrate 50 mg bid and carvedilol 25 mg bid had similar effects on 24-h heart rate. This result suggests that the degree of beta1-blockade produced by these two drugs in these doses is comparable and the superior survival effect of carvedilol compared to metoprolol seen in COMET is likely to be due to actions of carvedilol other than beta1-blockade. PMID- 16043408 TI - Left ventricular hypertrabeculation/non-compaction also occurs in nonagenarians. PMID- 16043409 TI - Utilization of evidence-based therapies for heart failure in the institutionalized elderly. AB - OBJECTIVES: Heart failure (HF) is a major source of morbidity and mortality in elderly populations. A significant proportion of the elderly with HF are living in long-term care facilities. Little is known about their management. The aim of this study was to evaluate the use of evidence-based therapies in institutionalized elderly patients with HF. DESIGN, SETTING AND PARTICIPANTS: Retrospective chart review conducted at 15 long-term care facilities in the Capital Health Region (Edmonton, Alberta). Residents > or =65 years of age with HF were identified using a pharmacy database. RESULTS: Overall prevalence of HF was 15% (313/2062 residents). Mean age was 87 years, median duration of residence was 1.8 years. Utilization of ACE-inhibitors, beta-blockers and spironolactone was 51%, 16% and 10%, respectively. Use of these medications was not significantly different between subgroups of those with and without contraindications to the therapies, different advance directive levels, gender or age. Sodium and fluid restricted diets were prescribed in only 11.0% and 3.8% of residents. Weight was not regularly monitored. Influenza and pneumococcal vaccination were administered to 60.4% and 81.2% of the residents. CONCLUSION: The use of evidence-based therapies in institutionalized elderly patients with HF is low, and unexplained by contraindications or advance directives. Efforts to increase the utilization of evidence-based therapies and improve monitoring are warranted. PMID- 16043410 TI - Revisiting Freund's incomplete adjuvant for vaccines in the developing world. AB - The rightful emphasis on collaboration between the public and private sectors in solving the world's health problems, especially those of the poor in the developing world, relies on industry developing new technologies with the support of government and foundations. However, certain products, despite their potential to have a great impact on disease in the developing world, will not be developed by industry because they carry the risk of lawsuits owing to possible severe adverse reactions--risks that are not counterbalanced by potential profit of products that are of limited use in the developed world. Freund's incomplete adjuvant (FIA) is one such example that is better developed in the public sector. In this article, we present the evidence that has led to our interest in FIA, evidence of its potential benefit in vaccines against blood-stage malaria, and the way forward to make safe, effective and affordable vaccines for malaria and other serious diseases in the developing world. PMID- 16043411 TI - Plasmodium parasite proteins and the infected erythrocyte. AB - Erythrocyte modification by malaria proteins is linked to both disease severity and infection. In this issue of Trends in Parasitology, Templeton and Deitsch, and Horrocks and Muhia discuss recent work identifying a host-targeting (HT) signal on malaria proteins. This signal predicts a secretome of 300-400 effectors for the human malaria parasite Plasmodium falciparum, vastly expanding the number of potential vaccine and drug targets. The HT signal seems to be distinct from known cellular transport signals, which suggests that it might be a novel eukaryotic secretion signal. PMID- 16043412 TI - Systems biology in malaria research. AB - A recent publication of genome and expression analyses of the murine parasites Plasmodium chabaudi chabaudi and Plasmodium berghei presents the state of the art in Plasmodium systems biology. By integrating genomics, transcriptomics and proteomics, the authors can classify and annotate genes by their expression profiles and can even detect evidence of posttranscriptional gene silencing in the murine malaria species. PMID- 16043413 TI - Genes, odours and the recognition of parasitized individuals by rodents. AB - Social recognition, whereby animals identify and recognize other individual conspecifics, is a crucial prerequisite for a wide range of social behaviours. There are relationships among social odours (chemical signals), parasite recognition and avoidance that are associated with hormonal, neural and genomic mechanisms in rodents. Rodents use social odours to: (i) distinguish between infected and uninfected individuals; (ii) recognize specific infected individuals; and (iii) avoid and display aversive responses to infected individuals. There are genomic correlates of this parasite recognition and avoidance in which genes expressing the neuropeptide oxytocin have roles. In this article, we provide a framework ("micronet") by which the genetic, hormonal and neural interactions associated with social behaviours and recognition and avoidance of parasitized individuals can be explored. PMID- 16043414 TI - Detection of IL-20 and its receptors on psoriatic skin. AB - The aim of this study was to investigate the expression of interleukin-20 (IL-20) and its receptors on psoriatic skin by immunohistochemical analysis and to evaluate the correlation of CD8-positive T lymphocytes with epidermal proliferation. Overexpression of IL-20 and its receptors was detected in the keratinocytes of the lesional skin of psoriasis and spongiotic dermatitis. The expression pattern of IL-20 spreads throughout the whole layer of epidermis, while IL-19 was expressed in up to three or four layers suprabasally. The serum level of IL-20 in psoriatic patients was significantly lower than that in healthy controls. IL-20 upregulated KGF transcripts on CD8-positive T cells. We hypothesize that overexpression of IL-20 is correlated with keratinocyte proliferation that acts through their receptor complex expressed by keratinocytes themselves. Furthermore, IL-20 can stimulate CD8-positive lymphocytes to produce KGF, which may contribute to sustaining the hyperproliferative status of the keratinocytes. PMID- 16043415 TI - Semi-automatic boundary detection to improve reporting of regional left ventricular function. AB - AIMS: The reporting of regional left ventricular function is based on subjective assessment of endocardial motion and thickening and has a significant learning curve. We hypothesized that the use of an semi-automatic boundary detection system generating images with superimposed moving endocardial borders and a fixed end-diastolic reference border could improve the reporting of regional function. METHODS: We obtained 58 resting contrast images of 15 patients and using a new boundary detection system (Quamus), generated images with superimposed endocardial borders. The contrast images, images with additional Quamus borders and Quamus borders alone were assessed by two level 1 and two level 2 echocardiographers. They scored regional function and results were compared to two level 3 experienced stress echocardiography readers. RESULTS: The addition of borders improved the agreement of level 1 echocardiographers (weighted Kappa increased from 0.55 to 0.64) but did not change for level 2 echocardiographers (0.63 to 0.64) and has the potential to be a useful training tool. PMID- 16043416 TI - The Serbian QOLIE-AD-48: translation, cultural adaptation, and preliminary psychometric evaluation. AB - PURPOSE: The goals of the work described in this article were to translate and adapt into Serbian the Quality of Life in Epilepsy Inventory for Adolescents (QOLIE-AD-48), and to evaluate the psychometric properties of the translation. METHODS: Translation/backtranslation, cultural adaptation, and pretesting were performed first. For validity, internal consistency, and sensitivity testing, the Serbian version was administered to a sample of 67 adolescents with epilepsy. RESULTS: All subscales (Epilepsy Impact, Memory/Concentration, Attitudes toward Epilepsy, Physical Functioning, Stigma, Social Support, School Behavior, and Health Perceptions) contributed significantly to the summary measure. Mean scores for the subscales ranged from 75.4 to 91.3, and showed a tendency to decrease as illness severity increased. Internal consistency reliability (Cronbach's alpha) was 0.92 for the entire questionnaire. CONCLUSIONS: The QOLIE-AD-48 Serbian version is a valid and reliable instrument for health-related quality-of-life assessment in youths with epilepsy. It can be administered in 15 minutes, and can be used in clinical practice, as well as in quality-of-life research. PMID- 16043417 TI - Suppression of obsessive-compulsive symptoms after epilepsy surgery. AB - We report two male patients with medically intractable epilepsy and obsessive compulsive disorder (OCD) symptoms. Both patients experienced remission of obsessive-compulsive symptoms after surgical treatment of epilepsy. Although the surgeries targeted different brain regions, the two patients had in common unilateral anterior cingulate cortex ablation. On the basis of these observations, we discuss the pathophysiology of OCD symptoms, emphasizing the role of corticosubcortical pathways in their genesis. Our data suggest that surgeries that affect neural loops associated with obsessive-compulsive symptoms can lead to an improvement of OCD; however, the structures responsible for this effect cannot be conclusively determined. PMID- 16043418 TI - Use of the Personality Assessment Inventory as an efficacious and cost-effective diagnostic tool for nonepileptic seizures. AB - Video electroencephalographic monitoring (VEEG) is considered the "gold standard" for making the differential diagnosis between epileptic seizures (ES) and nonepileptic seizures (NES), but is a costly, time-consuming procedure and not readily available in all communities. Of the various diagnostic techniques and measures that have been used, the Personality Assessment Inventory (PAI) has shown promise as an effective psychological screening tool to aid in the differential diagnosis of ES/NES. Using VEEG results as the outcome measure, this study examined the diagnostic effectiveness of the PAI in a group of adults with treatment-refractory seizures. Results indicated that, on psychological screening, patients with NES endorse significantly greater functional consequences of their seizure-like episodes than participants with ES. A "NES Indicator" score, calculated from the PAI Somatization subscales, provided a sensitivity of 84% and specificity of 73% for the diagnosis of NES versus ES. The PAI appears to be a useful screening tool prior to hospital admission for VEEG. PMID- 16043419 TI - Angiogenesis and cardioprotection after TNFalpha-inducer-Tolpa Peat Preparation treatment in rat's hearts after experimental myocardial infarction in vivo. AB - The aim of the presented work was to evaluate whether short subcutaneous (s.c.) administration of TNFalpha-inducer-Tolpa Peat Preparation (TPP or TPP batch 0210) modulates the process of ischemic remodeling and spontaneous angiogenesis after experimental myocardial infarction (MI) in rats in vivo. The results obtained using three complementary and correlative methods: histological studies, Proliferating Cell Nuclear Antigen (PCNA) reaction and Lymphocytes Induced Angiogenesis (LIA) test showed a clear pro-angiogenic and cardioprotective effect of TPP administration after experimental MI. TPP batch 0210 should be considered as an angiogenesis stimulating factor and consecutively as a cardioprotective preventing development of ischemic cardiomyopathy after MI in rats. It might possibly be used as an adjunct to conventional therapy of coronary artery disease, including late phase after myocardial infarction or ischemic cardiomyopathy. PMID- 16043420 TI - Characteristics of Flo11-dependent flocculation in Saccharomyces cerevisiae. AB - The FLO11-encoded flocculin is required for a variety of important phenotypes in Saccharomyces cerevisiae, including flocculation, adhesion to agar and plastic, invasive growth, pseudohyphae formation and biofilm development. We present evidence that Flo11p belongs to the Flo1-type class of flocculins rather than to the NewFlo class. Both Flo1-type and NewFlo yeast flocculation are inhibited by mannose. NewFlo flocculation, however, is also inhibited by several other carbohydrates including glucose, maltose and sucrose. These differences have in at least one case been shown to reflect differences in the structure of the carbohydrate-binding site of the flocculins. We report that Flo11p-dependent flocculation is inhibited by mannose, but not by glucose, maltose or sucrose. Furthermore, Flo11p contains a peptide sequence highly similar to one that has been shown to characterise Flo1-type flocculins. Further characterisation of the properties of Flo11p-dependent flocculation revealed that it is dependent on calcium, occurs only at cell densities greater than 1 x 10(8) ml(-1), and only occurs at acidic pH. PMID- 16043421 TI - Voltammetric investigation of cytochrome c on gold coated with a self-assembled glutathione monolayer. AB - The direct, reversible electrochemistry of horse-heart cytochrome c (cyt. c) was realized on a self-assembled glutathione (GSH) monolayer modified Au electrode. The voltammetric responses of cyt. c on GSH/Au electrode were found to be affected by pH during the electrode modification, metal ions and surfactants. Using potassium ferricyanide [K4Fe(CN)6] as a probe, these effects on the voltammetric responses of cyt. c were characterized by electrochemical methods. It was found that the pH during the electrode modification, metallic ions and surfactants changed GSH monolayer's charge state and the conformation on the electrode surface, and resulted in the influence on the voltammetric responses of cyt. c. The experimental results provided us information to understand the mechanism of the interfacial electron transfer of electrode-protein, as well as the electron transfer of cyt. c in life system. PMID- 16043422 TI - Electrochemistry and voltammetry of procaine using a carbon nanotube film coated electrode. AB - A new rapid, convenient and sensitive electrochemical method is described for the determination of procaine in pharmaceutical preparations, based on the unique properties of a multi-wall carbon nanotube (MWNT) thin film. The electrochemical behavior of procaine at the MWNT film-coated glassy carbon electrode (GCE) was investigated in detail, showing that the MWNT-coated GCE exhibits electrocatalytic activity to the oxidation of procaine because of the significant peak current enhancement and the lowering of oxidation overpotential. Furthermore, the mechanism for the oxidation of procaine at the MWNT-coated GCE was also studied. Finally, various experimental parameters such as solution pH value, the amount of MWNT, accumulation conditions and scan rate were optimized for the determination of procaine, and a new method with detection limit of 2 x 10(-7) mol/L was developed for procaine determination. This newly proposed method was successfully demonstrated with procaine hydrochloride injection. PMID- 16043423 TI - Differentiating human keratinocytes are deficient in p53 but retain global nucleotide excision repair following ultraviolet radiation. AB - Terminally differentiating keratinocytes constitute the predominant cell type within the skin epidermis and play an important role in the overall photobiology of human skin following ultraviolet radiation. However, the DNA repair capacity of differentiating keratinocytes is unclear, and little is known regarding how such repair activity is regulated in these cells. We systematically compared the global genomic nucleotide excision repair response of cultured undifferentiated human keratinocytes to those that were allowed to differentiate in 1.2 mM Ca(2+), in some cases supplemented with phorbol ester or Vitamin C. Differentiated cells ceased replication and expressed typical markers of differentiation. Following ultraviolet radiation, keratinocytes that were differentiated up to 12 days removed cyclobutane pyrimidine dimers and pyrimidine(6,4)pyrimidone photoproducts from the global genome as efficiently as undifferentiated cells. However, following the onset of calcium-induced differentiation, basal levels of p53 were nearly undetectable by 12 days of differentiation when global repair activity was unaffected. Following ultraviolet radiation, induction of p53 following ultraviolet radiation was abrogated by 6 days of calcium-induced differentiation. Basal levels of mRNA encoding the DNA damage recognition proteins, XPC and DDB2, were relatively insensitive to differentiation and p53 levels. However, following ultraviolet radiation, inductions of mRNA encoding the DNA damage recognition proteins, DDB2 and XPC, were differentially affected by differentiation. Rapid loss of DDB2 mRNA induction was associated with differentiation, while XPC mRNA induction diminished more slowly with differentiation. These results indicate that human keratinocytes preserve global nucleotide excision repair as well as expression of genes encoding key DNA damage recognition proteins well into the terminal differentiation process, perhaps using mechanisms other than p53. PMID- 16043424 TI - The Mre11/Rad50/Xrs2 complex and non-homologous end-joining of incompatible ends in S. cerevisiae. AB - In Saccharomyces cerevisiae, the Mre11/Rad50/Xrs2 (MRX) complex plays important roles in both homologous and non-homologous pathways of DNA repair. In this study, we investigated the role of the MRX complex and its enzymatic functions in non-homologous repair of DNA ends containing incompatible end structures. Using a plasmid transformation assay, we found that mre11 and rad50 null strains are extremely deficient in joining of incompatible DNA ends. Expression of the nuclease-deficient Mre11 mutant H125N fully complemented the mre11 strain for joining of mismatched ends in the absence of homology, while a mutant of Rad50 deficient in ATP-dependent activities exhibited levels of end-joining similar to a rad50 deletion strain. Although the majority of non-homologous end-joining (NHEJ) products isolated did not contain microhomologies, introduction of an 8bp microhomology at mismatched ends resulted in microhomology-mediated joining in all of the products recovered, demonstrating that a microhomology exerts a dominant effect on processing events that occur during NHEJ. Nuclease-deficient Mre11p was less efficient in promoting microhomology-mediated end-joining in comparison to its ability to stimulate non-microhomology-mediated events, suggesting that Mre11p influences, but is not essential for, microhomology mediated repair. When the linearized DNA was transformed in the presence of an intact homologous plasmid to facilitate gap repair, there was no decrease in NHEJ products obtained, suggesting that NHEJ and homologous repair do not compete for DNA ends in vivo. These results suggest that the MRX complex is essential for joining of incompatible ends by NHEJ, and the ATP-dependent activities of Rad50 are critical for this process. PMID- 16043425 TI - Genetic determinants of upper airway structures that predispose to obstructive sleep apnea. AB - Genetic factors are thought to play an important role in human development. Recent data indicate that obstructive sleep apnea may have a genetic basis. Sleep apnea is a very common disorder with significant cardiovascular and neurophysiologic morbidity. The pathogenesis of sleep apnea is related to a reduction in the size of the upper airway. The reduction in airway size is secondary to increased adipose tissue (enlargement of the parapharyngeal fat pads), alterations in craniofacial structure (reduction in mandibular size) and enlargement of the surrounding soft tissue structures (tongue, lateral pharyngeal walls). Genetic factors are one of the factors that have been proposed to mediate the size of each of these anatomic risk factors for sleep apnea. Recent evidence is accumulating about the genetic loci for these structural risk factors that predispose to the development of obstructive sleep apnea. PMID- 16043426 TI - High performance liquid chromatography-electrospray ionization mass spectrometric determination of tolterodine tartrate in human plasma. AB - A selective and sensitive high performance liquid chromatography-electrospray ionization mass spectrometry method has been developed for the determination of tolterodine tartrate in human plasma. With oxybutynin as internal standard, tolterodine tartrate was extracted from plasma with n-hexane: isopropanol (95:5, v/v). The organic layer was evaporated and the residue was redissolved in mobile phase comprised of acetonitrile-water (10 mM CH3COONH4, pH 3.0)=50:50 (v/v). An aliquot of 10 microl was chromatographically analyzed on a prepacked Shimadzu Shim-pack VP-ODS C18 column (150 mmx2.0 mm I.D.) by means of selected-ion monitoring (SIM) mode mass spectrometry. Standard curves were linear (r=0.9993) over the concentration range of 0.1-30.0 ng/ml and had good accuracy and precision. The within- and between-batch precisions were within 10% relative standard deviation. The limit of detection (LOD) was 0.05 ng/ml. The validated LC ESI-MS method has been used successfully to study tolterodine tartrate pharmacokinetic, bioavailability and bioequivalence in 20 healthy male volunteers. PMID- 16043427 TI - Determination of metformin in human plasma by high-performance liquid chromatography. AB - A simple, selective and sensitive high-performance liquid chromatographic method with spectrophotometric detection was developed for the determination of antihyperglycemic agent metformin in human plasma using a novel sample extraction procedure. Liquid-liquid extraction of metformin and ranitidine (as internal standard) from plasma samples was performed with 1-butanol/n-hexane (50:50, v/v) in alkaline condition followed by back-extraction into diluted acetic acid. Chromatography was carried out using a silica column (250 mmx4.6 mm, 5 microm) under isocratic elution with acetonitrile-40 mM aqueous sodium dihydrogen phosphate (25:75, v/v), pH 6. The limit of quantification (LOQ) was 15.6 ng/ml and the calibration curves were linear up to 2000 ng/ml. The mean absolute recoveries for metformin and internal standard using the present extraction procedure were 98 and 95%, respectively. The intra- and inter-day coefficient of variation and percent error values of the assay method were all less than 8.3%. PMID- 16043428 TI - RhBMP-7 improves survival and eruption in a growing tooth avulsion trauma model. AB - Long-term loss of avulsed and replanted teeth is a frequent clinical problem. Bone morphogenetic protein-7 (BMP 7) induces cementogenesis in periodontitis associated periodontal ligament (PDL) defects. This study's aim was to assess the utility of rhBMP 7 in a tooth avulsion trauma model in growing individuals. Immature primary incisors of 12 minipigs were extracted. PDL and cementum were removed either partially (group 1: 4 mm2 [n=28 teeth]; group 2: 16 mm2 [n=26 teeth]) or totally (group 3 [n=26 teeth]). 500 microg rhBMP 7/g collagen matrix was applied to the teeth from one side while the corresponding teeth on the contralateral side served as controls (split mouth model). After an experimental period of 4 months, microradiography, fluorescence and light microscopy of nondecalcified sections were performed. All teeth of group 1 survived and all teeth of group 3 were lost, whether rhBMP-7 was applied or not. In group 2, nine out of ten teeth survived when rhBMP-7 was applied and four out of ten teeth were lost when rhBMP-7 was not applied. In the presence of rhBMP-7, eruption of teeth in group 2 was significantly improved (difference [median]: 5 mm, P<0.05, n=6). Even though there was a tendency towards increased deposition rates of cementum under rhBMP-7, this difference was not significant (Wilcoxon: P>0.05, ANOVA: P=0.002; n=6/group). In conclusion, rhBMP-7 improved survival rates and eruption of replanted teeth in growing individuals. No adverse effects were seen. Based on the present results, future clinical trials appear to be warranted. PMID- 16043429 TI - A novel binding site in collagen type III for integrins alpha1beta1 and alpha2beta1. AB - Previously identified high affinity integrin-binding motifs in collagens, GFOGER and GLOGER, are not present in type III collagen. Here, we first characterized the binding of recombinant I domains from integrins alpha(1) and alpha(2) (alpha(1)I and alpha(2)I) to fibrillar collagen types I-III and showed that each I domain bound to the three types of collagens with similar affinities. Using rotary shadowing followed by electron microscopy, we identified a high affinity binding region in human type III collagen recognized by alpha(1)I and alpha(2)I. Examination of the region revealed the presence of two sequences that contain the critical GER motif, GROGER and GAOGER. Collagen-like peptides containing these two motifs were synthesized, and their triple helical nature was confirmed by circular dichroism spectroscopy. Experiments show that the GROGER-containing peptide was able to bind both alpha(1)I and alpha(2)I with high affinity and effectively inhibit the binding of alpha(1)I and alpha(2)I to type III and I collagens, whereas the GAOGER-containing peptide was considerably less effective. Furthermore, the GROGER-containing peptide supported adhesion of human lung fibroblast cells when coated on a culture dish. Thus, we have identified a novel high affinity binding sequence for the collagen-binding integrin I domains. PMID- 16043430 TI - Expression profile of the genes for rice cation/H+ exchanger family and functional analysis in yeast. AB - We identified five cation/H(+) exchangers (CAX) from rice, and phylogenetically divided them into two clusters. Gene expression and absolute amounts of mRNA in different organs were analyzed by real-time PCR. OsCAX1a showed high expression in almost all organs. OsCAX1b and OsCAX1c were detected in a limited number of organs and their expression levels were very low. The mRNA levels of OsCAX2 and OsCAX3 varied with the organ. OsCAXs were heterologously expressed in yeast to characterize the ion transport activity. All exchangers, except for OsCAX2, conferred tolerance to calcium. OsCAX1a and OsCAX3 conferred tolerance to manganese. The diversity of expression sites and substrates suggest the broad range function of CAX. PMID- 16043431 TI - Cochleata: getting to the root of legume nodules. AB - The homeotic mutant of Pisum sativum, cochleata, has stipules replaced by alternative leaf components, abnormal flowers and reduced fertility. Although the root system dry weight, root lengths and nodule numbers of cochleata are similar to those of its wild type, the nodulation phenotype of the mutant is unique. The nodules typically dichotomously branch and multiple callus and root structures emerge from their meristems. These nodule-roots incorporate a peripheral vascular bundle of the nodule into their own central vascular cylinder. Both the nodules and roots of the hybrid structures appear functional. Roles for COCHLEATA in development are discussed. PMID- 16043432 TI - The IRREGULAR TRICHOME BRANCH loci regulate trichome elongation in Arabidopsis. AB - The proper control of cell expansion is vital to plant development. It is responsible for shaping individual cells and, together with cell division, it plays a lead role in shaping plant organs. Much of the underlying mechanism by which plant cells expand anisotropically is not understood. We are taking a genetic approach to cell expansion by isolating mutants that affect the branching pattern of Arabidopsis trichomes. Here we report the identification of four new loci that control trichome morphogenesis. These loci were named the IRREGULAR TRICHOME BRANCH (ITB) loci because of the deleterious effects on branch position and length in the mutants. Our analysis of branch expansion in itb mutants shows that the ITB genes act as positive regulators of branch elongation, and that the branch position defects are caused by altered expansion of the trichome stalk. The itb mutations display synergistic effects in double mutant combinations with certain branch number mutations, suggesting that the ITB genes also play key roles in branch initiation. These results demonstrate that the ITB genes are key regulators of anisotropic cell expansion in trichomes. PMID- 16043433 TI - An open randomized study comparing disulfiram and acamprosate in the treatment of alcohol dependence. AB - AIMS: To compare the efficacy of acamprosate (ACP) and disulfiram (DSF) for preventing alcoholic relapse in routine clinical practice. METHODS: One hundred alcoholic men with family members who would encourage medication compliance and accompany them for follow-up were randomly allocated to 8 months of treatment with DSF or ACP. Weekly group psychotherapy was also available. The psychiatrist, patient, and family member were aware of the treatment prescribed. Alcohol consumption, craving, and adverse events were recorded weekly for 3 months and then fortnightly. Serum gamma glutamyl transferase was measured at the start and the end of the study. RESULTS: At the end of the trial, 93 patients were still in contact. Relapse (the consumption of >5 drinks/40 g of alcohol) occurred at a mean of 123 days with DSF compared to 71 days with ACP (P = 0.0001). Eighty-eight per cent of patients on DSF remained abstinent compared to 46% with ACP (P = 0.0002). However, patients allocated to ACP had lower craving than those on DSF (P = 0.002). CONCLUSION: DSF is superior to ACP for preventing relapse in alcohol dependent men with good family support. Further comparisons between these two drugs in different treatment settings and populations are warranted. PMID- 16043434 TI - Female surgeons' alcohol use: a study of a national sample of norwegian doctors. AB - AIMS: This study was designed to describe the alcohol use by female surgeons and the hazards of their drinking habits for them compared with the habits of female doctors from non-surgical specialities, and with those of their male colleagues in surgery, and to identify the variables associated with hazardous drinking. METHODS: The data were collected in 2000 from a representative national sample of 1120 Norwegian doctors. Alcohol use was measured using a modified version of the Alcohol Use Disorders Identification Test. A score of 9 or more was used as an indicator of hazardous drinking. RESULTS: Female surgeons compared with female non-surgeons had tendencies for more frequent moderate alcohol consumption accompanied by more frequent consumption of larger amounts of alcohol, and a significantly higher rate of hazardous drinking (18 vs 7.6%). Being a surgeon (OR = 1.7, 95% CI 1.2-2.4), male (OR = 2.7, 1.7-4.1) and aged 45 years or over (OR = 1.5, 1.1-2.2) were significant predictors of hazardous drinking. With separate gender analyses, being a surgeon was a significant predictor for both females (OR = 2.8, 1.2-6.6) and males (OR = 1.5, 1.0-2.3). CONCLUSION: Female surgeons practising in Norway drink more frequently and more hazardously than other female doctors. There are a number of possible explanations for this. Surgical culture may be an important factor. PMID- 16043435 TI - Attitudes of Swedish general practitioners and nurses to working with lifestyle change, with special reference to alcohol consumption. AB - AIMS: To explore the attitudes of Swedish general practitioners (GPs) and nurses to secondary alcohol prevention (early identification of, and intervention for, alcohol-related problems) and compare it to their attitudes to other important lifestyle behaviours such as smoking, stress, exercise, and overweight. METHODS: An adjusted version of The WHO Collaborative Study Questionnaire for General Practitioners was posted to all GPs and nurses in the County of Skaraborg, Sweden; 68 GPs and 193 nurses responded. RESULTS: The importance of drinking alcohol moderately, counselling skills on reducing alcohol consumption and perceived current effectiveness in helping patients change lifestyle behaviours ranked lower than working with all the other lifestyle behaviours. The nurses rated their potential effectiveness in helping patients change lifestyle higher than that of GPs for all the lifestyle behaviours. Nurses receiving more alcohol related education had more positive attitudes than nurses with less education. For alcohol, the GPs assessed their role adequacy, role legitimacy and motivation higher than that of the nurses. The main obstacles for the GPs to carry out alcohol intervention were lack of training in counselling for reducing alcohol consumption, time constraints, and the fact that the doctors did not know how to identify problem drinkers who have no obvious symptoms of excess consumption. CONCLUSION: GPs and the nurses estimated their alcohol-related competence as lower than working with many other health-related lifestyles. These results can be explained by lack of practical skills, lack of training in suitable intervention techniques, and unsupportive working environments. All these elements must be considered when planning secondary alcohol prevention programs in primary health care. PMID- 16043436 TI - Voluntary ethanol intake increases extracellular acetylcholine levels in the ventral tegmental area in the rat. AB - AIMS: Concurrent use of ethanol and nicotine (tobacco) is often seen in human beings. In previous animal experiments, we have demonstrated that nicotinic acetylcholine receptors, especially alpha-conotoxin MII and mecamylamine sensitive receptors located in the ventral tegmental area may be involved in the stimulatory, dopamine enhancing, and rewarding effects of ethanol in rodents. Ethanol may exert these effects via direct interaction with nicotinic acetylcholine receptors and/or indirectly via enhancement of extracellular acetylcholine levels in the ventral tegmental area. The present experiments investigated a possible indirect effect of ethanol in stimulating the mesoaccumbal dopamine system. METHODS: Neurochemical effects of voluntary ethanol intake on extracellular ventral tegmental acetylcholine and accumbal dopamine levels were measured by means of in vivo microdialysis with a two-probe approach in freely moving rats. RESULTS: Obtained data indicate that voluntary ethanol intake ( approximately 0.7 g/kg/h) leads to an increase of extracellular acetylcholine levels in the ventral tegmental area, and an almost time-locked increase of dopamine levels in the nucleus accumbens. A positive correlation between the ventral tegmental acetylcholine levels and ethanol intake as well as preference was also observed. CONCLUSION: The present results suggest that voluntary ethanol intake enhances extracellular ventral tegmental acetylcholine that may interact with nicotinic acetylcholine receptors, possibly alpha conotoxin MII sensitive receptors, localized in the ventral tegmental area that subsequently may stimulate dopamine overflow in the nucleus accumbens. PMID- 16043437 TI - The perisperm-endosperm envelope in Cucumis: structure, proton diffusion and cell wall hydrolysing activity. AB - BACKGROUND: and Aims The envelope surrounding the embryo in cucurbit seed, which consists of a single layer of live endosperm cells covered by lipid- and callose rich layers, is reported to show semi-permeability and also to act as the primary barrier to radicle emergence. Structure, development and permeability of the envelope and activity of cell wall hydrolases during germination of cucumber and muskmelon seeds were investigated. METHODS: Sections of seeds were stained with aniline blue and Sudan III. Proton diffusion and endo-beta-mannanase activity were detected by tissue printing. A gel-diffusion assay was performed to quantify endo-beta-mannanase activity, while the activity of beta-glucanase was determined with laminarin as the substrate and glucose formation measured using the GOD-POD method. KEY RESULTS: The lipid layer differentiated during seed development in cucumber in the epidermis of a multilayered nucellus, whereas the callose layer appeared to develop outside the endosperm cell layer. Accordingly, the envelope has been called the perisperm-endosperm (PE) envelope. Chloroform treatment of seeds, which resulted in a substantial reduction in Sudan staining of the lipid layer, also enhanced the permeability of the PE envelope to 2,3,5 triphenyltetrazolium chloride. Proton diffusion occurred when the PE envelopes from seeds had their inner surface in contact with bromocresol purple-containing agarose gels, but not when their outer surface was in contact. Substantial endo beta-mannanase activity was present in the caps of the PE envelopes, whereas a marked increase in beta-glucanase activity was observed in radicles prior to germination. CONCLUSIONS: The lipid layer seems to contribute to the semi permeability of the PE envelope. The diffusion of protons might create an acidic environment conducive to the activity of cell wall hydrolases, namely endo-beta mannanase (EC 3.2.1.78) and beta-glucanase [beta(1-->3)glucanohydrolase; EC 3.2.1.6], which, in turn, may play a role in the weakening of the PE envelope necessary for the protrusion of the radicle in cucumber and muskmelon seeds. PMID- 16043438 TI - High genetic diversity and clonal growth in relict populations of Olea europaea subsp. laperrinei (Oleaceae) from Hoggar, Algeria. AB - BACKGROUND AND AIMS: The Laperrine's olive (Olea europaea subsp. laperrinei) is an endemic tree from Saharan massifs. Its populations have substantially regressed since the Pleistocene and are presently distributed in a fragmented habitat. Long-term persistence of this taxon is uncertain and programmes of preservation have to be urgently implemented. To define a conservation strategy, the genetic diversity and breeding system of this tree have to be investigated. METHODS: One hundred and eleven ramets were prospected in the laperrinei populations from the Tamanrasset region, southern Algeria. Genetic polymorphism was revealed at nuclear and chloroplast DNA (cpDNA) microsatellite loci allowing a comparative assessment of the genetic diversity of laperrinei and Mediterranean populations based on bi-parental and maternal markers. Additionally, nuclear microsatellite markers enabled the genotypes to be identified unambiguously. KEY RESULTS: Based on nuclear microsatellite data, the total diversity was high (Ht=0.61) in laperrinei populations and similar to that observed in western Mediterranean populations. A substantial cpDNA diversity (Ht=0.19) was also observed. Genetically identical ramets originated from the same stump (which can cover >80 m2) were identified in each population. Sixteen per cent of genets exhibited more than one ramet. In addition, several cases of somatic mutations were unambiguously revealed in distinct ramets stemming from the same stump. CONCLUSIONS: These data show that highly isolated and small laperrinei populations are able to maintain a high genetic diversity. This supports the existence of relict trees persisting for a very long time (probably since the last humid transition, 3000 years ago). It is proposed that the very long persistence associated with an asexual multiplication of highly adapted trees could be a strategy of survival in extreme conditions avoiding a mutational meltdown due to reproduction in reduced populations. PMID- 16043439 TI - Phenotypic plasticity of growth trajectory and ontogenic allometry in response to density for eucalyptus hybrid clones and families. AB - BACKGROUND: and Aims Response to density is a crucial aspect of the ecology of trees in forests and plantations. Few studies have investigated the genetics of plasticity in response to density for growth traits such as height and circumference through development. METHODS: Two experiments were carried out in the field, the first with full-sib families of Eucalyptus urophylla x E. grandis hybrids, and the second with clones of E. tereticornis x E. grandis hybrids planted across a range of densities (625, 1111 and 2500 trees ha-1). Height, circumference and stem taper were measured through development in both experiments. Variance components were estimated and a repeated measure approach for plasticity and three different methods were used to compare the variance covariance matrix across densities. KEY RESULTS: Genetic variance was significantly different from zero but the density x genotype interaction was significant only for clone experiments at the adult stage. Significant plasticity for three traits in both experiments was found. In the clone experiments, a significant clone x time x density interaction was found, suggesting that plasticity for growth and stem form is under genetic control. In both experiments, density did not affect environmental correlation, which remained high throughout tree development. The impact of density on genetic correlation was marked in the clone experiment, with a reduced value at lower density, but was not observed in the family trial. The differences between clones and family are mainly explained by the distribution of genetic variation within and among genotypes. CONCLUSIONS: The results suggest that plasticity for growth traits and form of tropical Eucalyptus species is under genetic control and that the environment changes genetic co-variation through ontogeny. The findings confirm that a tree population with a narrow genetic basis (represented by clones) is sensitive to a changing environment, whereas a population with a broader genetic basis (full-sib family here) exhibits a more stable reaction. PMID- 16043440 TI - Correlates of Human Herpesvirus-8 DNA detection among adults in Italy without Kaposi sarcoma. AB - BACKGROUND: The presence of Human Herpesvirus-8 (HHV8) DNA is predictive of Kaposi sarcoma (KS) among patients with HIV-associated or iatrogenic immunosuppression. However, correlates of HHV8-DNA detection in the general population remain undefined. METHODS: We assessed correlates of HHV8-DNA detection among Italian adults without KS who had antibodies against HHV8-latent nuclear antigen by immunofluorescence assay. HHV8-K6 DNA sequences were detected in peripheral blood mononuclear cells using TaqMan PCR. RESULTS: Of the 158 subjects 26 (16.5%) had detectable HHV8-DNA [median copies/million cells, 53; (13 2128)]. Adjusted for age, sex, and laboratory, HHV8-DNA was detected more frequently in participants with >7 total residents in the childhood home [OR = 3.7 (1.5-9.1)], >2 younger siblings [OR = 2.6 (1.1-6.5)], and current cardiovascular [OR = 3.6 (1.3-9.7)] or renal [OR = 3.1 (1.2-8.0)] disease. Excluding the participants using immune modulating drugs, HHV8-DNA was more frequent among those with low red blood cells (RBC) [<4.5 10(6)/microl; OR = 5.3 (1.7-16.2)], slightly elevated mean corpuscular volume [>92 microm3/red cell; OR = 2.8 (1.0-7.8)], and mild thrombocytopenia [<151 K/microl; OR = 5.6 (1.9-16.3)]. CONCLUSIONS: Presence of HHV8-DNA in elderly Italians is associated with childhood crowding, low RBCs, and platelets, perhaps indicating roles for early infection and chronic inflammation. These risk factors are the first to be reported for non-immunosuppressed HHV8-seropositive adults. PMID- 16043441 TI - How many genes underlie the occurrence of common complex diseases in the population? AB - BACKGROUND: Most common human diseases are due to complex interactions among multiple genetic variants and environmental risk factors. There is debate over whether variants of a relatively small number of genes, each with weak or modest individual effects, account for a large proportion of common diseases in the population, or whether a large number of rare variants with large effects underlie genetic susceptibility to these diseases. It is not clear how many genes are necessary to account for an appreciable population-attributable fraction of these diseases. METHODS: In this analysis, we estimated the number of disease susceptibility genes needed to account for varying population attributable fractions of a common complex disease, taking into account the genotype prevalence, risk ratios for individual genes, and the model of gene-gene interactions (additive or multiplicative). RESULTS: Very large numbers of rare genotypes (e.g. those with frequencies of 1 per 5000 or less) are needed to explain 50% of a common disease in the population, even if the individual risk ratios are large (RR = 10-20). On the other hand, only approximately 20 genes are usually needed to explain 50% of the burden of a disease in the population if the predisposing genotypes are common (> or = 25%), even if the individual risk ratios are relatively small (RR = 1.2-1.5). CONCLUSIONS: Our results suggest that a limited number of disease susceptibility genes with common variants can explain a major proportion of common complex diseases in the population. Our findings should help focus the search for common genetic variants that provide the most important predispositions to complex human diseases. PMID- 16043442 TI - Increased use of emergency services by older people after health screening. AB - BACKGROUND: evaluation of the 'Keep Well At Home' (KWAH) Project in West London indicated that a programme of screening persons aged 75 and over had not reduced rates of emergency attendances and admissions to hospital. However, coverage of the target population was incomplete. The present analysis addresses 'efficacy' whether individuals who completed the screening protocol as intended did subsequently use Accident & Emergency (A&E) services less often. METHODS: the target population was divided into five groups, depending on whether an individual had completed none, one or both phases of screening, and whether deviations from the protocol related to incomplete coverage or refusal to participate further. We ascertained use of emergency services before screening and for up to 3 years afterwards by linkage of records from KWAH to those of local A&E Departments. Patterns of emergency care were examined as crude rates and, via proportional hazards models, after adjustment for available confounders. RESULTS: there was an increase of 51% (95% CI 22-86%) in the crude rate of emergency admissions in the year after first-phase screening compared with the 12 months before assessment. This was most obvious in individuals deemed at high risk who also underwent the second-phase assessment (adjusted hazard ratio relative to individuals not 'at risk'=2.33; 95% CI 1.59-3.42). CONCLUSIONS: the available data do not allow us to distinguish between several possible explanations for the paradoxical increase in use of emergency services. However, what seem to be sensible policies do not necessarily have their intended effects when implemented in practice. PMID- 16043443 TI - A multi-centre European study of factors affecting the discharge destination of older people admitted to hospital: analysis of in-hospital data from the ACMEplus project. AB - OBJECTIVES: to examine the relationship between seven predictor variables (recorded on Day 3 of hospital admission) and discharge destination in non elective medical patients aged 65+ years. DESIGN: prospective cohort. SETTING: eight centres in six European countries. PREDICTOR VARIABLES: age, gender, living alone, physical function (three categories based on Barthel Index), cognition (Katzman's orientation-memory-concentration test), main body system affected (based on International Classification of Diseases), number of geriatric giants (GGs) involved in the referral (a GG being a problem with falling, mobility, continence or cognition). MAIN OUTCOME MEASURES: discharge destination (by Day 90) in three categories: 'HOMESAME' (return to previous residence), 'INSTIN90' (discharge to alternative residence or still in hospital at 90 days), 'DEADINHO' (death in hospital), RESULTS: in 1,626 patients, discharge destination was HOMESAME in 84.7%, DEADINHO in 8.9% and INSTIN90 in 6.4%. Mean duration of stay was 17.7 days, median 12. Univariate analyses showed a statistically significant relationship between all seven predictor variables and discharge destination. Physical function was the best single predictor with a seven-fold difference in adverse outcome rates between the best and worst categories. On multiple logistic regression, significant predictor variables were as follows. (i) For DEADINHO: physical function, cognition, gender; (ii) for INSTIN90: physical function, living alone, GGs, age, gender. Multiple linear regression identified physical function, GGs and living alone as predictors of loge length of stay. CONCLUSION: case-mix systems to compare risk-adjusted hospital outcome in older medical patients need to incorporate information about physical function, cognition and presenting problems in addition to diagnosis. PMID- 16043444 TI - Vitamin D concentrations among people aged 65 years and over living in private households and institutions in England: population survey. AB - BACKGROUND: vitamin D deficiency among older people results in poor bone and muscle health and an increased risk of fractures. In the UK, government initiatives and the launch of the Osteoporosis Strategy have been in place since 1998, highlighting the importance of adequate levels of vitamin D for its prevention. The aim of this analysis is to assess vitamin D status and examine associations of deficiency with risk factors among older people in England. METHODS: a valid vitamin D sample was obtained from 1,766 informants as part of the Health Survey for England (HSE) 2000, a nationally representative survey of people aged 65 and over living in institutions and private households in England. RESULTS: among both men and women in institutions, the prevalence of vitamin D deficiency was higher and mean serum vitamin D levels were significantly lower than among those in private households. Regression analyses showed that women were more likely to be vitamin D deficient than men (odds ratio (OR) 2.1) and deficiency was associated with limiting longstanding illness (OR 3.57), manual social classes (OR 2.4), poor general health (OR 1.92) and body mass index<25 kg/m2 (OR 2.02), and was 67% more likely among informants in the winter/autumn. Overall, the results show no significant improvements in vitamin D status in comparison to earlier National Diet and Nutrition Survey (NDNS) results. CONCLUSION: vitamin D deficiency exists at worrying levels among those aged 65 years and over. Further action is needed to alert health professionals about the risks related to vitamin D deficiency and extend the provision of prevention and treatment programmes targeted to those in need. PMID- 16043445 TI - Sexuality in older adults: behaviours and preferences. AB - BACKGROUND: while much has been written about adult sexuality, relatively little is available about the sexuality of older people. Available literature often does not discuss specific sexual behaviours and includes predominantly married, better educated, mostly young old. OBJECTIVE: the purpose of this study was to assess a sample of lower-income older adults, about whom there is limited information, to describe a full range of sexual behaviours and to identify the degree to which they are satisfied with their sexual activities. METHODS: subjects were 179 people (60 and older) who were residents of subsidised independent-living facilities, recruited during a lecture or in public areas in the building. Thirteen of 179 were excluded due to age. Most were white (82%), living alone (83%) and female (63%). RESULTS: overall, the majority reported to have had physical and sexual experiences in the past year such as touching/holding hands (60.5%), embracing/hugging (61.7%) and kissing (57%) daily to at least once a month; mutual stroking, masturbation and intercourse were experienced 'not at all' by 82% or more. For all activities except masturbation, participants wanted to participate in sexual activities more often than they did. The most important barrier to sexual activity was lack of a partner. Self-reported health was related to sexual activities wanted, with age also related to some preferences. CONCLUSIONS: most of the elderly surveyed want to maintain a sexual relationship which includes touching and kissing, and they would like to have more sexual experiences than they have accessible. Further studies are needed. PMID- 16043446 TI - Chronic pain as perceived by older people: a qualitative study. AB - BACKGROUND: the practical issues confronting older people who suffer chronic pain may not be tackled in a pain clinic setting and little is known of their strategies for coping. They seem to have little or no information on how to improve the quality of their lives or on resources available to them. AIM: the aim of this study was to ascertain from older people the practical, physical and psychosocial limitations they faced because of chronic pain, and the strategies they used to deal with them. METHOD: a qualitative approach to generating data was chosen using a Grounded Theory approach and unstructured interviews. Sixty three people ranging from 60 to 87 years of age participated in the study. Audio tapes were transcribed verbatim. The material was coded and collapsed into themes. RESULTS: two main themes emerged: (i) the desire for independence and control; and (ii) adaptation to a life with chronic pain. The valuing of independence is in line with previous findings. With only three exceptions none of the participants were certain how or where to get help with practical issues and so they lived in fear of loss of their independence. Several sub-categories formed the theme of adaptation. These were acceptance and non-acceptance, pacing oneself, helping other people, the use of prayer and 'looking good and feeling good'. When independence and control is effective, older people may adapt better to chronic pain. CONCLUSION: understanding chronic pain sufferers from their own perspective may have important clinical implications. The interview data informed the development of a booklet designed to meet the needs of older patients with chronic pain. PMID- 16043447 TI - Kinetic folding mechanism of PDZ2 from PTP-BL. AB - PDZ domains represent a large family of protein-interaction modules associated with a variety of unrelated proteins with different functions. We report a complete characterization of the kinetic folding mechanism of a fluorescent variant of PDZ2 from PTP-BL, investigated under a variety of different experimental conditions. For this purpose, we engineered a fluorescent variant of this protein Y43W (called pseudo-wild-type, pWT43). The results suggest the presence of a high-energy intermediate in the folding of PDZ2, as revealed by a pronounced non-linear dependence of the unfolding rate constant on denaturant concentration. Such an intermediate may or may not be detectable depending on the experimental conditions, giving rise to apparent two-state folding under stabilizing conditions (e.g. in the presence of sodium sulfate). Interestingly, even under these conditions, three-state folding can be restored by selectively destabilizing the native-like rate-limiting barrier by one specific mutation (V44A). Finally, we show that data taken on pWT43 under different experimental conditions (e.g. different pH values from 2.1 to 8.0 or in the presence of a stabilizing salt) and also data on a site-directed conservative mutant can be rationalized in terms of a simple reaction scheme involving a single set of intermediates and transition states. PMID- 16043448 TI - Engineering allosteric protein switches by domain insertion. AB - Domain insertion is proving to be an effective way to construct hybrid proteins exhibiting switch-like behavior. In this strategy, two existing domains, the first exhibiting a signal recognition function and the second containing the function to be modulated, are fused such that the recognition of the signal by the first domain is transmitted to the second domain, thereby modulating its activity. Recent directed evolution experiments indicate that the structural space comprised of the recombination of unrelated protein domains may be rich in switching behavior, particularly when the circular permutation of domains is also employed. This bodes well for potential basic science, sensing and therapeutic applications of molecular switches. PMID- 16043449 TI - Combined pharmacologic treatment with clopidogrel and statin for patients with acute coronary syndrome: is there a survival advantage? PMID- 16043450 TI - Ethanol breaks dormancy of the potato tuber apical bud. AB - Growing potato tubers or freshly harvested mature tubers have a dormant apical bud. Normally, this dormancy is spontaneously broken after a period of maturation of the tuber, resulting in the growth of a new sprout. Here it is shown that in in vitro-cultured growing and maturing tubers, ethanol can rapidly break this dormancy and re-induce growth of the apical bud. The in vivo promoter activity of selected genes during this secondary growth of the apical bud was monitored, using luciferase as a reporter. In response to ethanol, the expression of carbohydrate-storage, protein-storage, and cell division-related genes are rapidly down-regulated in tuber tissue. It was shown that dormancy was broken by primary but not by secondary alcohols, and the effect of ethanol on sprouting and gene expression in tuber tissue was blocked by an inhibitor of alcohol dehydrogenase. By contrast, products derived from alcohol dehydrogenase activity (acetaldehyde and acetic acid) did not induce sprouting, nor did they affect luciferase reporter gene activity in the tuber tissue. Application of an inhibitor of gibberellin biosynthesis had no effect on ethanol-induced sprouting. It is suggested that ethanol-induced sprouting may be related to an alcohol dehydrogenase-mediated increase in the catabolic redox charge [NADH/(NADH+NAD+)]. PMID- 16043451 TI - Transcript enrichment of Nod factor-elicited early nodulin genes in purified root hair fractions of the model legume Medicago truncatula. AB - This article describes an efficient procedure to study Nod factor-induced gene expression in root hairs of the model legume Medicago truncatula. By developing an improved method of fracturing frozen root hairs, it has been possible to obtain a highly purified root hair fraction from M. truncatula seedlings yielding sufficient RNA for real-time quantitative RT-PCR expression analysis. After Nod factor treatment it was possible to detect up to 100-fold increases of MtENOD11 and pMtENOD11-gus transcript levels in root hair RNA. This corresponds to 5-7 fold higher induction levels than for entire root tissue preparations. Furthermore, the use of these enriched RNA samples has revealed for the first time a very significant induction (30-fold) of the MtENOD40 gene in root hairs in response to Nod factors. It is concluded that the rapid and convenient procedure described here will be particularly useful for detecting tissue-specific low level gene expression in root hairs responding to Rhizobium Nod factors or other exogenous signals. PMID- 16043452 TI - GAL4-GFP enhancer trap lines for genetic manipulation of lateral root development in Arabidopsis thaliana. AB - Lateral root development occurs throughout the life of the plant and is responsible for the plasticity of the root system. In Arabidopsis thaliana, lateral root founder cells originate from pericycle cells adjacent to xylem poles. In order to study the mechanisms of lateral root development, a population of Arabidopsis GAL4-GFP enhancer trap lines were screened and two lines were isolated with GAL4 expression in root xylem-pole pericycle cells (J0121), i.e. in cells competent to become lateral root founder cells, and in young lateral root primordia (J0192). These two enhancer trap lines are very useful tools with which to study the molecular and cellular bases of lateral root development using targeted gene expression. These lines were used for genetic ablation experiments by targeting the expression of a toxin-encoding gene. Moreover, the molecular bases of the enhancer trap expression pattern were characterized. These results suggest that the lateral-root-specific GAL4 expression pattern in J0192 is due to a strong enhancer in the promoter of the LOB-domain protein gene LBD16. PMID- 16043453 TI - NaCl alleviates polyethylene glycol-induced water stress in the halophyte species Atriplex halimus L. AB - Atriplex halimus L. is a C4 xero-halophyte species well adapted to salt and drought conditions. To collect information on the physiological impact of low salt levels on their water-stress resistance, seedlings were exposed for 6 d to nutrient solution containing either 0% or 15% polyethylene glycol 10,000 (PEG), in the presence or in the absence of 50 mM NaCl. Similar experiments were performed with one PEG-resistant and one PEG-sensitive selected cell line exposed for 50 d to 0% or 15% PEG on standard Linsmaier and Skoog (LS) medium, on LS medium supplemented with 50 mM NaCl, or on Na+-free medium. NaCl mitigated the deleterious impact of PEG on growth of both whole plants and PEG-sensitive cell lines and improved the ability of stressed tissues to perform osmotic adjustment (OA). Water stress reduced CO2 net assimilation rates quantified in the presence of high CO2 and low O2 levels (A), stomatal conductance and transpiration, but NaCl improved water use efficiency of PEG-treated plants through its positive effect on A values, especially in young leaves. PEG increased the internal Na+ concentration. The resistant cell line accumulated higher concentration of Na+ than the PEG-sensitive one. The complete absence of Na+ in the medium endangered the survival of both cell lines exposed to PEG. Although Na+ by itself contributed only for a small part to OA, NaCl induced an increase in proline concentration and stimulated the synthesis of glycinebetaine in response to PEG in photosynthetic tissues. Soluble sugars were the main contributors to OA and increased when tissues were simultaneously exposed to PEG and NaCl compared with PEG alone, suggesting that Na+ may influence sugar synthesis and/or translocation. PMID- 16043454 TI - The Arabidopsis mutant eer2 has enhanced ethylene responses in the light. AB - By screening for ethylene response mutants in Arabidopsis, a novel mutant, eer2, was isolated which displays enhanced ethylene responses. On a low nutrient medium (LNM) light-grown eer2 seedlings showed a significant hypocotyl elongation in response to low levels of 1-amino-cyclopropane-1-carboxylate (ACC), the precursor of ethylene, compared with the wild type, indicating that eer2 is hypersensitive to ethylene. Treatment with 1-MCP (1-methylcyclopropene), a competitive inhibitor of ethylene signalling, suppressed this hypersensitive response, demonstrating that it is a bona fide ethylene effect. By contrast, roots of eer2 were less sensitive than the wild type to low concentrations of ACC. The ethylene levels in eer2 did not differ from the wild type, indicating that ethylene overproduction is not the primary cause of the eer2 phenotype. In addition to its enhanced ethylene response of hypocotyls, eer2 is also affected in the pattern of senescence and its phenotype depends on the nutritional status of the growth medium. Furthermore, linkage analysis of eer2 suggests that this mutant defines a new locus in ethylene signalling. PMID- 16043455 TI - Relationships between xylem sap constituents and leaf conductance of well-watered and water-stressed maize across three xylem sap sampling techniques. AB - Many different techniques have been used for xylem sap collection, but few direct comparisons of techniques have been conducted and few comparisons have been based on comprehensive analyses of xylem sap. Moreover, the suitability of extraction techniques for use on plants grown under water-stress conditions has not been addressed. Xylem sap was extracted from both well-watered and water-stressed Zea mays plants using three different techniques. The main aim was to determine how the extraction method altered the correlations between sap constituents and stomatal conductance in order to determine which relationships change with extraction technique. A 'root pressure' technique was the simplest method of extracting large volumes of sap, but the low sap delivery rates altered the composition of sap. Two pressurization techniques that varied in the position from which sap was collected were tested. The pressurization techniques allowed for the control of delivery rates that influence sap constituent concentrations. The position from which xylem sap was collected on the plant was also found to be important. All three techniques produced consistent correlations between ABA and chloride delivery rates and changes in stomatal conductance, suggesting that each technique could be applied to identify certain putative xylem-borne signals. PMID- 16043456 TI - Modification of MRI criteria for multiple sclerosis in patients with clinically isolated syndromes. AB - BACKGROUND: The McDonald criteria include MRI evidence for dissemination in space and dissemination in time for the diagnosis of multiple sclerosis in young adult patients who present with clinically isolated syndromes (CIS) typical of the disease. Although a major advance, the criteria have limited sensitivity for making an early diagnosis. OBJECTIVE: To compare the performance of McDonald criteria and modified McDonald criteria for dissemination in space and time for assessing the development of clinically definite multiple sclerosis. METHODS: McDonald criteria were modified using the combination of a less stringent definition for dissemination in space and allowing a new T2 lesion per se after three months as evidence for dissemination in time. Modified and McDonald criteria were applied in 90 CIS patients at baseline and at three month follow up scans. RESULTS: Both criteria were highly specific (>90%) but the modified criteria were more sensitive (77% v 46%) and more accurate (86% v 73%). CONCLUSIONS: These modified criteria should be evaluated in other CIS cohorts. PMID- 16043457 TI - Functional neuroimaging and presenting psychiatric features in frontotemporal dementia. AB - BACKGROUND: Frontotemporal dementia (FTD) is a behavioural syndrome caused by degeneration of the frontal and anterior temporal lobes. Behavioural disturbances include psychiatric features. Whether patients with FTD present with psychiatric features varies with the initial neuroanatomical variability of FTD. OBJECTIVE: To identify presenting psychiatric changes not part of diagnostic criteria of FTD and contrast them with the degree of hemispheric asymmetry and frontal and temporal hypoperfusion on single photon emission computed tomography (SPECT) imaging. METHODS: 74 patients who met consensus criteria for FTD were evaluated at a two year follow up. All had brain SPECT on initial presentation. Results of an FTD psychiatric checklist were contrasted with ratings of regional hypoperfusion. RESULTS: The regions of predominant hypoperfusion did not correlate with differences on FTD demographic variables but were associated with presenting psychiatric features. Dysthymia and anxiety were associated with right temporal hypoperfusion. "Moria" or frivolous behaviour also occurred with temporal lobe changes, especially on the right. The only significant frontal lobe feature was the presence of a peculiar physical bearing in association with right frontal hypoperfusion. CONCLUSIONS: Patients with FTD may present with psychiatric changes distinct from the behavioural diagnostic criteria for this disorder. Early temporal involvement is associated with frivolous behaviour and right temporal involvement is associated with emotional disturbances. In contrast, those with right frontal disease may present with alterations in non verbal behaviour. PMID- 16043458 TI - Crucial roles of mesodermal cell lineages in a murine embryonic stem cell-derived in vitro liver organogenesis system. AB - Recent studies in the field of regenerative medicine have exploited the pluripotency of embryonic stem (ES) cells to generate a variety of cell lineages. However, the target has always been only a single lineage, which was isolated from other differentiated cell populations. In the present study, we selected sublines with a high capability for differentiation to contracting cardiomyocytes and also produced germ-line chimeric mice from a parent ES line. We also succeed in establishing embryoid bodies prepared from the ES cells that differentiated into not only hepatocytes but also at least two mesodermal lineages: cardiomyocytes that supported liver development and endothelial cells corresponding to sinusoids. This allowed the development of an in vitro system using murine ES cells that approximated the events of liver development in vivo. The expression of albumin was significantly higher in cardiomyocytes that had arisen in differentiated ES cells than in those that had not. Our in vitro system for liver organogenesis consists of a blood/sinusoid vascular-like network and hepatocyte layers and shows higher levels of hepatic function, such as albumin production and ammonia degradation, than hepatic cell lines and primary cultures of murine adult hepatocytes. This innovative system will lead to the development of second-generation regenerative medicine techniques using ES cells and is expected to be useful for the development of bioartificial liver systems and drug metabolism assays. PMID- 16043459 TI - Voltage-sensitive and ligand-gated channels in differentiating neural stem-like cells derived from the nonhematopoietic fraction of human umbilical cord blood. AB - Fetal cells with the characteristics of neural stem cells (NSCs) can be derived from the nonhematopoietic fraction of human umbilical cord blood (HUCB), expanded as a nonimmortalized cell line (HUCB-NSC), and further differentiated into neuron like cells (HUCB-NSCD); however, the functional and neuronal properties of these cells are poorly understood. To address this issue, we used whole-cell patch clamp recordings, gene microarrays, and immunocytochemistry to identify voltage gated channels and ligand-gated receptors on HUCB-NSCs and HUCB-NSCDs. Gene microarray analysis identified genes for voltage-dependent potassium and sodium channels and the neurotransmitter receptors acetylcholine (ACh), gamma aminobutyric acid (GABA), glutamate, glycine, 5-hydroxytryptamine (5-HT), and dopamine (DA). Several of these genes (GABA-A, glycine and glutamate receptors, voltage-gated potassium channels, and voltage-gated sodium type XII alpha channels) were not expressed in the HUCB mono-nuclear fraction (HUCB-MC), which served as a starting cell population for HUCB-NSC. HUCB-NSCD acquired neuronal phenotypes and displayed an inward rectifying potassium current (Kir) and an outward rectifying potassium current (I(K+)). Kir was present on most HUCB-NSCs and HUCB-NSCDs, whereas I(K+) was present only on HUCB-NSCDs. Many HUCB-NSCDs were immunopositive for glutamate, glycine, nicotinic ACh, DA, 5-HT, and GABA receptors. Kainic acid (KA), a non-N-methyl-D-asparate (NMDA) glutamate-receptor agonist, induced an inward current in some HUCB-NSCDs. KA, glycine, DA, ACh, GABA, and 5-HT partially blocked Kir through their respective receptors. These results suggest that HUCB-NSCs differentiate toward neuron-like cells, with functional voltage- and ligand-gated channels identified in other neuronal systems. PMID- 16043460 TI - Cartilage engineering from ovine umbilical cord blood mesenchymal progenitor cells. AB - We aimed to determine whether three-dimensional (3D) cartilage could be engineered from umbilical cord blood (CB) cells and compare it with both engineered fetal cartilage and native tissue. Ovine mesenchymal progenitor cells were isolated from CB samples (n=4) harvested at 80-120 days of gestation by low density fractionation, expanded, and seeded onto polyglycolic acid scaffolds. Constructs (n=28) were maintained in a rotating bioreactor with serum-free medium supplemented with transforming growth factor-beta1 for 4-12 weeks. Similar constructs seeded with fetal chondrocytes (n=13) were cultured in parallel for 8 weeks. All specimens were analyzed and compared with native fetal cartilage samples (n=10). Statistical analysis was by analysis of variance and Student's t test (p<.01). At 12 weeks, CB constructs exhibited chondrogenic differentiation by both standard and matrix-specific staining. In the CB constructs, there was a significant time-dependent increase in extracellular matrix levels of glycosaminoglycans (GAGs) and type-II collagen (C-II) but not of elastin (EL). Fetal chondrocyte and CB constructs had similar GAG and C-II contents, but CB constructs had less EL. Compared with both hyaline and elastic native fetal cartilage, C-II and EL levels were, respectively, similar and lower in the CB constructs, which had correspondingly lower and similar GAG levels than native hyaline and elastic fetal cartilage. We conclude that CB mesenchymal progenitor cells can be successfully used for the engineering of 3D cartilaginous tissue in vitro, displaying select histological and functional properties of both native and engineered fetal cartilage. Cartilage engineered from CB may prove useful for the treatment of select congenital anomalies. PMID- 16043461 TI - Determination of thrombopoietin-derived peptides recognized by both cellular and humoral immunities in healthy donors and patients with thrombocytopenia. AB - Thrombopoietin (TPO) is a cytokine that promotes megakaryocytopoiesis and thrombopoiesis and is considered a drug suitable for patients with thrombocytopenia. However, unexpected severe thrombocytopenia has developed in some healthy individuals participating in phase I clinical trials with a pegylated recombinant human megakaryocyte growth factor (PEG-rHuMGDF) that contained the first 163 amino acids of endogenous TPO, which resulted in hampering the further development of clinical trials. Autoimmune responses to PEG rHuMGDF, which cross-reacted with endogenous TPO, were suggested to be involved in this rare but severe adverse event, although the immunogenic epitopes have not yet been determined. To better understand the molecular basis of such autoimmune reactions, we investigated the reactivity of 18 TPO-derived peptides with HLA-A2 binding motifs to plasma and T cells, both from patients with thrombocytopenia (n=24) and from healthy donors (HDs) (n=24). Four peptides, including those possessing amino acids in receptor-binding sites, were preferentially reactive to plasma from at least 20% of the patients, whereas one peptide at position 101-109 was equally reactive to those of the patients and the HDs. Each of the five peptides had the ability to induce peptide-specific cytotoxic T lymphocytes (CTLs) in both groups, albeit with less frequency among the patients. More important, each of these five peptides had the ability to induce HLA-A2 restricted and peptide-specific CTL activity reactive to cells that produce TPO. These results may provide new insights to gain a better understanding of autoimmune reactions to TPO. PMID- 16043462 TI - Aquaporin-3 expression in human fetal airway epithelial progenitor cells. AB - Airway epithelium stem cells have not yet been prospectively identified, but it is generally assumed that both secretory and basal cells have the capacity to divide and differentiate. Previously, we developed a test for progenitor cells of the human airway epithelium, relying on the transplantation of fetal respiratory tissues into immunodeficient mice. In this study, we hypothesized that airway repopulating epithelial progenitors can be marked with surface antigens, and we screened an array of such candidate markers, including lectin ligands, the CD44 and CD166 adhesion molecules, and the aquaporin-3 (AQP3) water channel. We observed that AQP3 is selectively expressed on the surface of basal cells, allowing the separation by flow cytometry of AQP3+ basal cells and AQP3- ciliated and secretory cells. Functional evaluation of sorted cells in vivo showed that AQP3+ cells can restore a normal pseudostratified, mucociliary epithelium as well as submucosal glands. AQP3- cells are also endowed with a similar potential, although faster engraftment suggests their inclusion of more committed progenitors. These results show that stem cell candidates in the human tracheo bronchial mucosa can be positively selected with a novel marker but also, for the first time, that epithelial progenitors exist among both basal and suprabasal cell subsets within the human airway. PMID- 16043463 TI - The molecular perspective: double-stranded DNA breaks. PMID- 16043464 TI - 'Dynamic clamp' in cardiac electrophysiology. PMID- 16043466 TI - Hypodontia in orthodontically treated children. AB - The frequency of hypodontia in orthodontically treated children, both male and female, and the association between tooth type, the upper or lower arch, the affected side and Angle's classification were studied using interviews, oral, study cast and panoramic radiographic examinations of 212 patients with a mean age of 12 years 7 months. A hypodontia frequency of 11.3 per cent was found for the total sample. This was higher than the incidence of hypodontia reported in other studies of orthodontically treated children. The most frequently missing teeth were the maxillary lateral incisors, and maxillary and mandibular second premolars. The missing teeth were more often absent on the right (54.2 per cent) than on the left (45.8 per cent) side, in both males and females. One tooth was absent in 29.2 per cent of patients, two in 58.5 per cent, but seldom three or more. Orthodontic space closure was the treatment of choice in 87.5 per cent of the subjects. PMID- 16043467 TI - Long-term skeletal effects of mandibular symphyseal distraction osteogenesis. An implant study. AB - The purpose of this study was to investigate the long-term skeletal effects of mandibular symphyseal distraction osteogenesis (MSDO) with a tooth- and bone borne distraction device, analysed using the metallic implant method. The study sample comprised 20 patients between 15.8 and 25 years of age, with a mean age of 20.01 +/- 2.25 years at the start of treatment. In 12 subjects, titanium implants were inserted in the mandible to analyse mandibular skeletal changes in the short and long term. A custom-made intraoral, tooth- and bone-borne distractor was designed and used. After a latency period of 7 days, the distractor was activated twice daily, by a total amount of 1 mm. Postero-anterior (PA) cephalograms were obtained at the start of distraction and at the end of consolidation (94.95 +/- 5.79 days after surgery) and follow-up periods (21.5 +/- 4.6 months after consolidation). The data were analysed statistically using paired t-tests. The mean amount of screw activation was 8.10 +/- 1.68 mm. The inter-symphyseal and inter-molar implant distances and the bimolar width significantly increased during the consolidation period (P < 0.001) and were maintained at the end of the follow-up. On the other hand, the bicondylar width was markedly decreased (P < 0.05), while no significant skeletal changes were observed in bigonion and biantigonion widths, inter-ramal implant distance, or inter-ramal and implant angles at the end of the consolidation period. The long-term findings of this study indicate that MSDO provides an efficient and stable non-extraction treatment alternative, mainly by increasing the anterior mandibular skeletal and dental arches. PMID- 16043468 TI - A treatment difficulty index for unerupted maxillary canines. AB - The aim of this study was to produce a treatment difficulty index (TDI) for unerupted maxillary canines. Thirty treated cases, each with an unerupted unilateral maxillary canine, were graded by 14 consultant orthodontists in terms of perceived alignment difficulty, and the four main factors which had contributed to each grade were listed in order of importance. The relationship between the grade and the contributory factors was then examined using regression analysis, and weightings were derived. These were applied to each factor, in order to derive a difficulty score total for each case. Linear regression analysis of difficulty scores against consultant grades produced an R2 value of 54.7 per cent using the original equation and an R2 of 52.3 per cent using values rounded to the nearest half. Both indicated a moderate level of agreement between allocated difficulty grade and calculated difficulty scores. The index provides a useful treatment planning aid for the management of impacted maxillary canines. PMID- 16043469 TI - An in vitro comparison of the shear bond strength of a resin-reinforced glass ionomer cement and a composite adhesive for bonding orthodontic brackets. AB - The shear bond strength (SBS) of a light-cured, resin-reinforced glass ionomer and a composite adhesive in combination with a self-etching primer was compared after different setting times to evaluate when orthodontic wires could be placed. Additionally, the fracture site after debonding was assessed using the Adhesive Remnant Index (ARI). Eighty freshly extracted human premolars were used. Twenty teeth were randomly assigned to each of four groups: (1) brackets bonded with Transbond XT with a Transbond Plus etching primer and debonded within 5 minutes; (2) brackets bonded with Fuji Ortho LC and debonded within 5 minutes; (3) brackets bonded as for group 1 and debonded within 15 minutes; (4) brackets bonded as for group 2 and debonded within 15 minutes. The SBS of each sample was determined with an Instron machine. The mean SBS were, respectively: (1) 8.8 +/- 2 MPa; (2) 6.6 +/- 2.5 MPa; (3) 11 +/- 1.6 MPa and (4) 9.6 +/- 1.6 MPa. Interpolating the cumulative fracture probability by means of a Weibull analysis, the 10 per cent probabilities of fracture for the groups were found to be attained for shear stresses of 6.1, 3.1, 8.3 and 7.1 MPa, respectively. Chi square testing of the ARI scores revealed that the nature of the remnant did not vary significantly with time, but the type of bonding material could generally be distinguished in leaving more or less than 10 per cent of bonding material on the tooth. After debonding, the Transbond system was likely to leave adhesive on at least 10 per cent of the bonded area of the tooth. The present findings indicate that brackets bonded with either Transbond XT in combination with Transbond Plus etching primer and Fuji Ortho LC had adequate bond strength at 5 minutes and were even stronger 15 minutes after initial bonding. PMID- 16043470 TI - Tensile bond strength of brackets after antioxidant treatment on bleached teeth. AB - Various studies have reported a significant reduction in tensile bond strength of brackets when bonding is carried out immediately after bleaching. The purpose of this investigation was to determine the effect of an antioxidant agent on the tensile bond strength values of metal brackets bonded with composite resin to human enamel after bleaching with carbamide peroxide (CP). A total of 80 extracted premolar teeth were randomly divided into three bleaching groups of 10 per cent CP and an unbleached control group. The specimens in group 1 were bonded immediately after bleaching; group 2 were stored in an artificial saliva solution for 7 days after bleaching; group 3 were treated with 10 per cent sodium ascorbate, immediately before bonding, whereas the unbleached specimens in group 4 had no treatment before bonding. Tensile bond strengths were established in MPa. To evaluate the amount of resin left on the enamel surfaces after debonding, the adhesive remnant index (ARI) scores were used. The tensile bond strength data were analyzed with the Kruskal-Wallis test and pairwise comparisons were made by the Mann-Whitney U test at a significance level of P < 0.05. The brackets bonded immediately after bleaching revealed significantly lower tensile bond strengths than those of unbleached enamel (P = 0.000). No statistically significant differences in tensile bond strength were noted when the delayed-bonding (P = 6.000) and antioxidant-treated (P = 0.2757) groups were compared with the control group. The antioxidant treatment immediately after bleaching was effective in reversing the tensile bond strength of brackets. PMID- 16043471 TI - Alterations in gingival dimensions following rapid canine retraction using dentoalveolar distraction osteogenesis. AB - The aim of this clinical prospective study was to evaluate the alterations that occurred in the gingival dimensions of canine teeth following dentoalveolar distraction (DAD) during a 12 month follow-up period.The study sample comprised 36 maxillary canines of 18 growing or adult subjects with a mean age of 16.94 years (13.08-25.58 years) at the start of treatment. Full retraction of the canines was achieved in 10.36 +/- 1.93 days (range 8-14 days) at a rate of 0.8 mm/day using a custom-made intraoral rigid tooth-borne distraction device. Before surgery (pre-DAD), immediately after removal of the device (post-DAD), and at 1, 6, and 12 months post-DAD, the plaque index (PI), gingival index (GI), pocket depth (PD) and width of keratinized gingiva were recorded and the width of attached gingiva was calculated. The alterations in clinical measurements among different evaluation periods were analysed by Friedman and repeated measure ANOVA tests. There were significant differences between pre- and post-DAD for PD measurements for all sites, with the highest at the distal site. The palatal sites likewise showed significant differences at the 1, 6, and 12 month follow-up periods compared with the post-DAD period. The buccal sites showed no significant changes at any time point. The width of keratinized gingiva also showed no significant change during the follow-up period, while the width of attached gingiva was significant only between the pre- and post-DAD periods (P < 0.01). On the basis of the above findings, it could be concluded that DAD is an innovative technique with no unfavourable long-term effects on the gingival tissues of rapidly retracted canine teeth. PMID- 16043472 TI - Ranking facial attractiveness. AB - The first aim of this investigation was to assemble a group of photographs of 30 male and 30 female faces representing a standardized spectrum of facial attractiveness, against which orthognathic treatment outcomes could be compared. The second aim was to investigate the influence of the relationship between ANB differences and anterior lower face height (ALFH) percentages on facial attractiveness. The initial sample comprised standardized photographs of 41 female and 35 male Caucasian subjects. From these, the photographs of two groups of 30 male and 30 female subjects were compiled. A panel of six clinicians and six non-clinicians ranked the photographs. The results showed there to be a good level of reliability for each assessor when ranking the photographs on two occasions, particularly for the clinicians (female subjects r = 0.76-0.97, male subjects r = 0.72-0.94). Agreement among individuals within each group was also high, particularly when ranking facial attractiveness in male subjects (female subjects r = 0.57-0.84, male subjects r = 0.91-0.94). Antero-posterior (AP) discrepancies, as measured by soft tissue ANB, showed minimal correlation with facial attractiveness. However, a trend emerged that would suggest that in faces where the ANB varies widely from 5 degrees, the face is considered less attractive. The ALFH percentage also showed minimal correlation with facial attractiveness. However, there was a trend that suggested that greater ALFH percentages are considered less attractive in female faces, while in males the opposite trend was seen. Either of the two series of ranked photographs as judged by clinicians and non-clinicians could be used as a standard against which facial attractiveness could be assessed, as both were in total agreement about the most attractive faces. However, to judge the outcome of orthognathic treatment, the series of ranked photographs produced by the non-clinician group should be used as the 'standard' to reflect lay opinion. PMID- 16043473 TI - Soft tissue facial angles in Down's syndrome subjects: a three-dimensional non invasive study. AB - The aim of the present study was to obtain quantitative information concerning the three-dimensional (3D) arrangement of the facial soft tissues of subjects with Down's syndrome. The 3D co-ordinates of 50 soft tissue facial landmarks were recorded by an electromechanical digitizer in 17 male and 11 female subjects with Down's syndrome aged 12-45 years, and in 429 healthy individuals of the same age, ethnicity and gender. From the landmark co-ordinates, geometric calculations were obtained of several 3D facial angles: facial convexity in the horizontal plane (upper facial convexity, mid facial convexity including the nose, and lower facial convexity), mandibular corpus convexity in the horizontal plane, facial convexity including the nose, facial convexity excluding the nose, interlabial angle, nasolabial angle, angle of nasal convexity, left and right soft tissue gonial angles. Data were compared with that collected for the normal subjects by computing the z-scores. Facial convexity in the horizontal plane (both in the upper and mid facial third), facial convexity in the sagittal plane and the angle of nasal convexity were significantly (P < 0.05) increased (flatter) in subjects with Down's syndrome than in the normal controls. Both left and right soft tissue gonial angles were significantly reduced (more acute) in the Down's syndrome subjects. Subjects with Down's syndrome had a more hypoplastic facial middle third with reduced nasal protrusion, and a reduced lower facial third (mandible) than reference, normal subjects. PMID- 16043474 TI - Roentgen-cephalometric standards for a Swedish population. A longitudinal study between the ages of 5 and 31 years. AB - This study was performed to establish age- and gender-specific cephalometric normative data for a Swedish population. The material comprised 469 lateral cephalograms from two groups of subjects of Swedish origin between 5 and 31 years of age. All subjects (males and females) were diagnosed as 'normal' according to specified criteria and with no history of orthodontic treatment. Lateral cephalograms and body height measurements were taken at 5, 7, 10 and 13 years of age in one group (group E) (longitudinal follow-up). The same registrations were made in the other group (group U) at 7, 10 and13 years of age (mixed longitudinal type). Those 13 year olds were re-examined at 16, 19 and 31 years of age (longitudinal follow-up). No significant differences between age groups in the two samples were found. Therefore, subjects of the same age were combined. The study is hence regarded as a longitudinal follow-up from 5 to 31 years of age. The subjects were also grouped into dental development stages to widen the applicability of the cephalometric data.The craniofacial distances were constantly larger in males than in females, while no statistical differences as regards angular measurements were seen between genders. Distances as well as angular measurements varied with the different developmental periods. The results clearly verify that facial pattern changes existed during the observation period, with a growth acceleration of most distances between the 13 and 16 year recordings. PMID- 16043475 TI - Validation of two-dimensional measurements of root resorption craters on human premolars after 28 days of force application. AB - The aims of this study were to develop a three-dimensional (3D) mathematical model of a typical root resorption crater and to correlate two-dimensional (2D) surface area measurements to 3D volumetric measurements of root resorption craters created under light and heavy orthodontic forces. Data were obtained from a previous study of 36 first premolars from 16 subjects requiring extraction of these teeth as part of their orthodontic treatment. Buccal tipping forces of 25 or 225 g were applied for an experimental period of 28 days. After extraction, the samples were prepared for scanning electron microscopy (SEM) imaging, image processing and analysis. Surface area (2D) and volumetric (3D) measurements of all craters were obtained. A mathematical analysis of the 2D/3D relationship enabled the determination of an appropriate digital model for the shape, type and dimensions of resorption craters, which was also able to distinguish between a 'hemispheric' model versus a 'layered' model of craters. The results demonstrated that 2D and 3D measurements were strongly correlated (r = 0.991**). Within the light and heavy force groups, the measurements were also strongly correlated (r = 0.978** and r = 0.994**, respectively). For a 28 day experimental period, 2D measurements of root resorption craters were found to be as reliable as 3D measurements. PMID- 16043476 TI - The craniofacial complex in 47, XXX females. AB - A study of the craniofacial complex in four 47, XXX Finnish females, or females with an extra X chromosome, was carried out using cephalometric analysis comprising linear and angular measurements. The lengths of the anterior and posterior cranial bases, the calvarium, mandibular ramus and posterior and upper anterior face heights were found to be significantly shorter than in female controls, while the angles between the foraminal and clival planes, the mandibular plane and cranial base, the maxillary and occlusal planes, the maxillary and mandibular planes and the foraminal and mandibular planes, and also the gonial angle, were significantly enlarged. The present findings of reduced linear measurements, together with the results of studies on the craniofacial complex of 47, XXY and 47, XYY males, suggest dimensional variation between these groups from the promoting effect of an extra Y chromosome and the retarding effect of an extra X chromosome on craniofacial growth. PMID- 16043477 TI - Evolution of flexural rigidity according to the cross-sectional dimension of a superelastic nickel titanium orthodontic wire. AB - The choice of the most suitable orthodontic wire for each stage of treatment requires estimation of the forces generated. In theory, the selection of wire sequences should initially utilize a lower flexural rigidity; thus clinicians use smaller round cross-sectional dimension wires to generate lighter forces during the preliminary alignment stage. This assessment is true for conventional alloys, but not necessarily for superelastic nickel titanium (NiTi). In this case, the flexural rigidity dependence on cross-sectional dimension differs from the linear elasticity prediction because of the martensitic transformation process. It decreases with increasing deflection and this phenomenon is accentuated in the unloading process. This behaviour should lead us to consider differently the biomechanical approach to orthodontic treatment. The present study compared bending in 10 archwires made from NiTi orthodontics alloy of two cross-sectional dimensions. The results were based on microstructural and mechanical investigations. With conventional alloys, the flexural rigidity was constant for each wire and increased largely with the cross-sectional dimension for the same strain. With NiTi alloys, the flexural rigidity is not constant and the influence of size was not as important as it should be. This result can be explained by the non-constant elastic modulus during the martensite transformation process. Thus, in some cases, treatment can begin with full-size (rectangular) wires that nearly fill the bracket slot with a force application deemed to be physiologically desirable for tooth movement and compatible with patient comfort. PMID- 16043478 TI - Enamel loss at bond-up, debond and clean-up following the use of a conventional light-cured composite and a resin-modified glass polyalkenoate cement. AB - The aim of this study was to determine whether there was any difference in the degree of enamel loss at bond-up, debond and enamel clean-up when two different adhesive systems were tested and with four different methods of enamel clean-up. The adhesive systems were 37 per cent o-phosphoric acid with Transbond XT (group 1) and 10 per cent poly(acrylic acid) conditioner with Fuji Ortho LC (group 2). Using flattened enamel specimens, enamel loss at each stage was determined using a planer surfometer. These stages were: prior to treatment, at pumice prophylaxis, following enamel etching or conditioning and following enamel clean up. The four clean-up methods were a high-speed tungsten carbide bur, a slow speed tungsten carbide bur, an ultrasonic scaler and debanding pliers. The results, analysed using non-parametric tests, demonstrated that significantly more enamel loss occurred following the use of 37 per cent o-phosphoric acid than poly(acrylic acid) conditioner (P = 0.001). At debond and prior to clean-up, more adhesive remained on the enamel surface in group 1 than in group 2 (P = 0.005). During the subsequent enamel clean-up and with both adhesive systems, the least enamel loss occurred following the use of the slow-speed tungsten carbide bur and the greatest loss was seen with the ultrasonic scaler or high-speed tungsten carbide bur.Overall, the lowest enamel loss was observed with the poly(acrylic acid) conditioner and Fuji Ortho LC system (group 2) and where enamel clean-up was performed using the slow-speed tungsten carbide bur. PMID- 16043479 TI - A subjective comparison of two lingual bracket systems. AB - The purpose of this prospective, longitudinal study was to compare the influence of two lingual bracket systems on subjective oral comfort, speech, mastication and oral hygiene. Forty-two native speakers of standard German (32 females, 10 males; mean age 27.1 years, standard deviation 12.2) were enrolled and completed a standardized questionnaire directly before insertion of lingual brackets (T0), within 24 hours of bond-up (T1) and 3 months (+/- 1 week) later (T2). Eighteen of the patients were treated with prefabricated brackets (Ormco, seventh generation) (PB group) and 24 with customized brackets (Incognito) (CB group). While no significant intergroup differences were recorded at any of the times with respect to tongue position, conversation pattern, swallowing or oral hygiene, the CB group experienced significantly fewer tongue space restrictions, speech disturbances and impairments in chewing and biting than the PB group at T1 and T2. At T2, pressure sores, reddening or lesions to the tongue were recorded significantly less often in the CB group than in the PB group. This enhanced patient comfort in the CB group was attributed to the smaller dimensions of the customized brackets. This aspect could play a role in attracting more patients to lingual orthodontics in the future. Information given to the patient on the duration and extent of the restrictions associated with lingual orthodontics must be differentiated according to the bracket system used. PMID- 16043480 TI - Mammalian target of rapamycin (mTOR) signaling is required for a late-stage fusion process during skeletal myotube maturation. AB - Skeletal myogenesis is a well orchestrated cascade of events regulated by multiple signaling pathways, one of which is recently characterized by its sensitivity to the bacterial macrolide rapamycin. Previously we reported that the mammalian target of rapamycin (mTOR) regulates the initiation of the differentiation program in mouse C2C12 myoblasts by controlling the expression of insulin-like growth factor-II in a kinase-independent manner. Here we provide experimental evidence suggesting that a different mode of mTOR signaling regulates skeletal myogenesis at a later stage. In the absence of endogenous mTOR function in C2C12 cells treated with rapamycin, a kinase-inactive mTOR fully supports myogenin expression, but causes a delay in contractile protein expression. Myoblasts fuse to form nascent myotubes in the absence of kinase active mTOR, whereas the formation of mature myotubes by further fusion requires the catalytic activity of mTOR. Therefore, the two stages of myocyte fusion are molecularly separable at the level of mTOR signaling. In addition, our data suggest that a factor secreted into the culture medium is responsible for mediating the function of mTOR in regulating the late-stage fusion leading to mature myotubes. Furthermore, taking advantage of the unique features of cells stably expressing a mutant mTOR, we have performed cDNA microarray analysis to compare global gene expression profiles between mature and nascent myotubes, the results of which have implicated classes of genes and revealed candidate regulators in myotube maturation or functions of mature myotubes. PMID- 16043482 TI - Rabphilin localizes with the cell actin cytoskeleton and stimulates association of granules with F-actin cross-linked by {alpha}-actinin. AB - In endocrine cell, granules accumulate within an F-actin-rich region below the plasma membrane. The mechanisms involved in this process are largely unknown. Rabphilin is a cytosolic protein that is expressed in neurons and neuroendocrine cells and binds with high affinity to members of the Rab3 family of GTPases localized to synaptic vesicles and dense core granules. Rabphilin also interacts with alpha-actinin, a protein that cross-links F-actin into bundles and networks and associates with the granule membrane. Here we asked whether rabphilin, in addition to its granule localization, also interacts with the cell actin cytoskeleton. Immunofluorescence and immunoelectron microscopy show that rabphilin localizes to the sub-plasmalemmal actin cytoskeleton both in neuroendocrine and unspecialized cells. By using purified components, it is found that association of rabphilin with F-actin is dependent on added alpha-actinin. In an in vitro assay, granules, unlike endosomes or mitochondria, associate with F-actin cross-linked by alpha-actinin. Rabphilin is shown to stimulate this process. Rabphilin enhances by approximately 8-fold the granule ability to localize within regions of elevated concentration of cross-linked F-actin. These results suggest that rabphilin, by interacting with alpha-actinin, organizes the cell cytoskeleton to facilitate granule localization within F-actin-rich regions. PMID- 16043481 TI - RPGR-ORF15, which is mutated in retinitis pigmentosa, associates with SMC1, SMC3, and microtubule transport proteins. AB - Mutations in the retinitis pigmentosa GTPase regulator (RPGR) gene account for almost 20% of patients with retinitis pigmentosa. Most mutations are detected in alternatively spliced RPGR-ORF15 isoform(s), which are primarily but not exclusively expressed in the retina. We show that, in addition to the axoneme, the RPGR-ORF15 protein is localized to the basal bodies of photoreceptor connecting cilium and to the tip and axoneme of sperm flagella. Mass spectrometric analysis of proteins that were immunoprecipitated from the retinal axoneme-enriched fraction using an anti-ORF15 antibody identified two chromosome associated proteins, structural maintenance of chromosomes (SMC) 1 and SMC3. Using pulldown assays, we demonstrate that the interaction of RPGR with SMC1 and SMC3 is mediated, at least in part, by the RCC1-like domain of RPGR. This interaction was not observed with phosphorylation-deficient mutants of SMC1. Both SMC1 and SMC3 localized to the cilia of retinal photoreceptors and Madin-Darby canine kidney cells, suggesting a broader physiological relevance of this interaction. Additional immunoprecipitation studies revealed the association of RPGR-ORF15 isoform(s) with the intraflagellar transport polypeptide IFT88 as well as microtubule motor proteins, including KIF3A, p150Glued, and p50-dynamitin. Inhibition of dynein function by overexpressing p50 abrogated the localization of RPGR-ORF15 to basal bodies. Taken together, these results provide novel evidence for the possible involvement of RPGR-ORF15 in microtubule organization and regulation of transport in primary cilia. PMID- 16043483 TI - MEF2-dependent recruitment of the HAND1 transcription factor results in synergistic activation of target promoters. AB - HAND proteins are tissue-restricted members of the basic helix-loop-helix transcription factor family that play critical roles in cell differentiation and organogenesis including placental, cardiovascular, and craniofacial development. Nevertheless, the molecular basis underlying the developmental action of HAND proteins remains undefined. Within the embryo, HAND1 is first detected in the developing heart where it becomes restricted to the atrial and left ventricular compartments, a pattern identical to that of the Nppa gene, which encodes atrial natriuretic factor, the major secretory product of the heart. We hereby report that the cardiac atrial natriuretic factor promoter is directly activated by HAND1, making it the first known HAND1 transcriptional target. The action of HAND1 does not require heterodimerization with class I basic helix-loop-helix factors or DNA binding through E-box elements. Instead, HAND1 is recruited to the promoter via physical interaction with MEF2 proteins. MEF2/HAND1 interaction results in synergistic activation of MEF2-dependent promoters, and MEF2 binding sites are sufficient to mediate this synergy. MEF2 binding to DNA is not enhanced in the presence of HAND1. Instead, cooperativity likely results from corecruitment of co-activators such as CREB-binding protein. The related HAND2 protein can also synergize with MEF2. Thus, HAND proteins act as cell-specific developmental co-activators of the MEF2 family of transcription factors. These findings identify a novel mechanism for HAND action in the heart and provide a general paradigm to understand the mechanism of HAND action in organogenesis. PMID- 16043484 TI - Basis for selectivity of cationic antimicrobial peptides for bacterial versus mammalian membranes. AB - Novel cationic antimicrobial peptides typified by structures such as KKKKKKAAXAAWAAXAA-NH2, where X = Phe/Trp, and several of their analogues display high activity against a variety of bacteria but exhibit no hemolytic activity even at high dose levels in mammalian erythrocytes. To elucidate their mechanism of action and source of selectivity for bacterial membranes, phospholipid mixtures mimicking the compositions of natural bacterial membranes (containing anionic lipids) and mammalian membranes (containing zwitterionic lipids + cholesterol) were challenged with the peptides. We found that peptides readily inserted into bacterial lipid mixtures, although no insertion was detected in model "mammalian" membranes. The depth of peptide insertion into model bacterial membranes was estimated by Trp fluorescence quenching using doxyl groups variably positioned along the phospholipid acyl chains. Peptide antimicrobial activity generally increased with increasing depth of peptide insertion. The overall results, in conjunction with molecular modeling, support an initial electrostatic interaction step in which bacterial membranes attract and bind peptide dimers onto the bacterial surface, followed by the "sinking" of the hydrophobic core segment to a peptide sequence-dependent depth of approximately 2.5-8 A into the membrane, largely parallel to the membrane surface. Antimicrobial activity was likely enhanced by the fact that the peptide sequences contain AXXXA sequence motifs, which promote their dimerization, and possibly higher oligomerization, as assessed by SDS-polyacrylamide gel analysis and fluorescence resonance energy transfer experiments. The high selectivity of these peptides for nonmammalian membranes, combined with their activity toward a wide spectrum of Gram-negative and Gram-positive bacteria and yeast, while retaining water solubility, represent significant advantages of this class of peptides. PMID- 16043485 TI - Functional consequences of hypertrophic and dilated cardiomyopathy-causing mutations in alpha-tropomyosin. AB - To study the functional consequences of various cardiomyopathic mutations in human cardiac alpha-tropomyosin (Tm), a method of depletion/reconstitution of native Tm and troponin (Tn) complex (Tm-Tn) in cardiac myofibril preparations has been developed. The endogenous Tm-Tn complex was selectively removed from myofibrils and replaced with recombinant wild-type or mutant proteins. Successful depletion and reconstitution steps were verified by SDS-gel electrophoresis and by the loss and regain of Ca2+-dependent regulation of ATPase activity. Five Tm mutations were chosen for this study: the hypertrophic cardiomyopathy (HCM) mutations E62Q, E180G, and L185R and the dilated cardiomyopathy (DCM) mutations E40K and E54K. Through the use of this new depletion/reconstitution method, the functional consequences of these mutations were determined utilizing myofibrillar ATPase measurements. The results of our studies showed that 1) depletion of >80% of Tm-Tn from myofibrils resulted in a complete loss of the Ca2+-regulated ATPase activity and a significant loss in the maximal ATPase activity, 2) reconstitution of exogenous wild-type Tm-Tn resulted in complete regain in the calcium regulation and in the maximal ATPase activity, and 3) all HCM-associated Tm mutations increased the Ca2+ sensitivity of ATPase activity and all had decreased abilities to inhibit ATPase activity. In contrast, the DCM-associated mutations both decreased the Ca2+ sensitivity of ATPase activity and had no effect on the inhibition of ATPase activity. These findings have demonstrated that the mutations which cause HCM and DCM disrupt discrete mechanisms, which may culminate in the distinct cardiomyopathic phenotypes. PMID- 16043486 TI - Crystal structure of lipoate-protein ligase A from Escherichia coli. Determination of the lipoic acid-binding site. AB - Lipoate-protein ligase A (LplA) catalyzes the formation of lipoyl-AMP from lipoate and ATP and then transfers the lipoyl moiety to a specific lysine residue on the acyltransferase subunit of alpha-ketoacid dehydrogenase complexes and on H protein of the glycine cleavage system. The lypoyllysine arm plays a pivotal role in the complexes by shuttling the reaction intermediate and reducing equivalents between the active sites of the components of the complexes. We have determined the X-ray crystal structures of Escherichia coli LplA alone and in a complex with lipoic acid at 2.4 and 2.9 angstroms resolution, respectively. The structure of LplA consists of a large N-terminal domain and a small C-terminal domain. The structure identifies the substrate binding pocket at the interface between the two domains. Lipoic acid is bound in a hydrophobic cavity in the N-terminal domain through hydrophobic interactions and a weak hydrogen bond between carboxyl group of lipoic acid and the Ser-72 or Arg-140 residue of LplA. No large conformational change was observed in the main chain structure upon the binding of lipoic acid. PMID- 16043487 TI - Vacuolar processing enzyme is essential for mycotoxin-induced cell death in Arabidopsis thaliana. AB - Some compatible pathogens secrete toxins to induce host cell death and promote their growth. The toxin-induced cell death is a pathogen strategy for infection. To clarify the executioner of the toxin-induced cell death, we examined a fungal toxin (fumonisin B1 (FB1))-induced cell death of Arabidopsis plants. FB1-induced cell death was accompanied with disruption of vacuolar membrane followed by lesion formation. The features of FB1-induced cell death were completely abolished in the Arabidopsis vacuolar processing enzyme (VPE)-null mutant, which lacks all four VPE genes of the genome. Interestingly, an inhibitor of caspase-1 abolished FB1-induced lesion formation, as did a VPE inhibitor. The VPE-null mutant had no detectable activities of caspase-1 or VPE in the FB1-treated leaves, although wild-type leaves had the caspase-1 and VPE activities, both of which were inhibited by a caspase-1 inhibitor. gammaVPE is the most essential among the four VPE homologues for FB1-induced cell death in Arabidopsis leaves. Recombinant gammaVPE recognized a VPE substrate with Km = 30.3 microm and a caspase-1 substrate with Km = 44.2 microm, which is comparable with the values for mammalian caspase-1. The gammaVPE precursor was self-catalytically converted into the mature form exhibiting caspase-1 activity. These in vivo and in vitro analyses demonstrate that gammaVPE is the proteinase that exhibits a caspase-1 activity. We show that VPE exhibiting a caspase-1 activity is a key molecule in toxin-induced cell death. Our findings suggest that a susceptible response of toxin-induced cell death is caused by the VPE-mediated vacuolar mechanism similar to a resistance response of hypersensitive cell death (Hatsugai, N., Kuroyanagi, M., Yamada, K., Meshi, T., Tsuda, S., Kondo, M., Nishimura, M., and Hara Nishimura, I. (2004) Science 305, 855-858). PMID- 16043488 TI - Angiomotin regulates endothelial cell-cell junctions and cell motility. AB - We have previously identified angiomotin by its ability to bind to and mediate the anti-angiogenic properties of angiostatin. In vivo and in vitro data indicate an essential role of angiomotin in endothelial cell motility. Here we show that angiostatin binds angiomotin on the cell surface and provide evidence for a transmembrane model for the topology of both p80 and p130 angiomotin isoforms. Immunofluorescence analysis shows that angiomotin co-localized with ZO-1 in cell cell contacts in endothelial cells in vitro and in angiogenic blood vessels of the postnatal mouse retina in vivo. Transfection of p80 as well as p130 angiomotin in Chinese hamster ovary cells resulted in junctional localization of both isoforms. Furthermore, p130 angiomotin could recruit ZO-1 to actin stress fibers. The p130 but not p80 isoform could be coprecipitated with MAGI-1b, a component of endothelial tight junctions. Paracellular permeability, as measured by diffusion of fluorescein isothiocyanate-dextran, was reduced by p80 and p130 angiomotin expression with 70 and 88%, respectively, compared with control. Angiostatin did not have any effect on cell permeability but inhibited the migration of angiomotin-expressing cells in the Boyden chamber assay. We conclude that angiomotin, in addition to controlling cell motility, may play a role in the assembly of endothelial cell-cell junctions. PMID- 16043489 TI - Functional expression of the human HCN3 channel. AB - Hyperpolarization-activated, cyclic nucleotide-gated cation (HCN) channels underlie the inward pacemaker current, termed I(f)/I(h), in a variety of tissues. Many details are known for the HCN subtypes 1, 2, and 4. We now successfully cloned the cDNA for HCN3 from human brain and compared the electrophysiological properties of hHCN3 to the other three HCN subtypes. Overexpression of human HCN3 channels in HEK293 cells resulted in a functional channel protein. Similar to hHCN2 channels, hHCN3 channels are activated with a rather slow time constant of 1244 +/- 526 ms at -100 mV, which is a greater time constant than that of HCN1 but a smaller one than that of HCN4 channels. The membrane potential for half maximal activation V((1/2)) was -77 +/- 5.4 mV, and the reversal potential E(rev) was -20.5 +/- 4 mV, resulting in a permeability ratio P(Na)/P(K) of 0.3. Like all other HCNs, hHCN3 was inhibited rapidly and reversibly by extracellular cesium and slowly and irreversibly by extracellular applied ZD7288. Surprisingly, the human HCN3 channel was not modulated by intracellular cAMP, a hallmark of the other known HCN channels. Sequence comparison revealed >80% homology of the transmembrane segments, the pore region, and the cyclic nucleotide binding domain of hHCN3 with the other HCN channels. The missing response to cAMP distinguishes human HCN3 from both the well cAMP responding HCN subtypes 2 and 4 and the weak responding subtype 1. PMID- 16043490 TI - Genetic inhibition or activation of JNK1/2 protects the myocardium from ischemia reperfusion-induced cell death in vivo. AB - The c-Jun NH2-terminal kinase (JNK) branch of the mitogen-activated protein kinase signaling cascade has been implicated in the regulation of apoptosis in a variety of mammalian cell types. In the heart, disagreement persists concerning the role that JNKs may play in regulating apoptosis, since both pro- and antiapoptotic regulatory functions have been reported in cultured cardiomyocytes. Here we report the first analysis of cardiomyocyte cell death due to JNK inhibition or activation in vivo using genetically modified mice. Three separate mouse models with selective JNK inhibition were assessed for ventricular damage and apoptosis levels following ischemia-reperfusion injury. jnk1-/-, jnk2-/-, and transgenic mice expressing dominant negative JNK1/2 within the heart were each shown to have less JNK activity in the heart and less injury and cellular apoptosis in vivo following ischemia-reperfusion injury. To potentially address the reciprocal gain-of-function phenotype associated with sustained JNK activation, transgenic mice were generated that express MKK7 in the heart. These transgenic mice displayed elevated cardiac c-Jun kinase activity but, ironically, were also significantly protected from ischemia-reperfusion. Mechanistically, JNK inhibited mice showed increased phosphorylation of the proapoptotic factor Bad at position 112, whereas MKK7 transgenic mice showed decreased phosphorylation of this site. Collectively, these results underscore the complexity associated with JNK signaling in regulating apoptosis, such that sustained inhibition or activation both elicit cellular protection in vivo, although probably through different mechanisms. PMID- 16043491 TI - Canonical WNT signaling promotes osteogenesis by directly stimulating Runx2 gene expression. AB - Both activating and null mutations of proteins required for canonical WNT signaling have revealed the importance of this pathway for normal skeletal development. However, tissue-specific transcriptional mechanisms through which WNT signaling promotes the differentiation of bone-forming cells have yet to be identified. Here, we address the hypothesis that canonical WNT signaling and the bone-related transcription factor RUNX2/CBFA1/AML3 are functionally linked components of a pathway required for the onset of osteoblast differentiation. Our findings show that, in bone of the SFRP1 (secreted frizzled-related protein-1) null mouse, which exhibits activated WNT signaling and a high bone mass phenotype, there is a significant increase in expression of T-cell factor (TCF) 1, Runx2, and the RUNX2 target gene osteocalcin. We demonstrate by mutational analysis that a functional TCF regulatory element responsive to canonical WNT signaling resides in the promoter of the Runx2 gene (-97 to -93). By chromatin immunoprecipitation, recruitment of beta-catenin and TCF1 to the endogenous Runx2 gene is shown. Coexpression of TCF1 with canonical WNT proteins resulted in a 2-5 fold activation of Runx2 promoter activity and a 7-8-fold induction of endogenous mRNA in mouse pluripotent mesenchymal and osteoprogenitor cells. This enhancement was abrogated by SFRP1. Taken together, our results provide evidence for direct regulation of Runx2 by canonical WNT signaling and suggest that Runx2 is a target of beta-catenin/TCF1 for the stimulation of bone formation. We propose that WNT/TCF1 signaling, like bone morphogenetic protein/transforming growth factor beta signaling, activates Runx2 gene expression in mesenchymal cells for the control of osteoblast differentiation and skeletal development. PMID- 16043492 TI - Vitamin A inhibits pancreatic stellate cell activation: implications for treatment of pancreatic fibrosis. AB - BACKGROUND AND AIMS: Activated pancreatic stellate cells (PSCs) are implicated in the production of alcohol induced pancreatic fibrosis. PSC activation is invariably associated with loss of cytoplasmic vitamin A (retinol) stores. Furthermore, retinol and ethanol are known to be metabolised by similar pathways. Our group and others have demonstrated that ethanol induced PSC activation is mediated by the mitogen activated protein kinase (MAPK) pathway but the specific role of retinol and its metabolites all-trans retinoic acid (ATRA) and 9-cis retinoic acid (9-RA) in PSC quiescence/activation, or its influence on ethanol induced PSC activation is not known. Therefore, the aims of this study were to (i) examine the effects of retinol, ATRA, and 9-RA on PSC activation; (ii) determine whether retinol, ATRA, and 9-RA influence MAPK signalling in PSCs; and (iii) assess the effect of retinol supplementation on PSCs activated by ethanol. METHODS: Cultured rat PSCs were incubated with retinol, ATRA, or 9-RA for varying time periods and assessed for: (i) proliferation; (ii) expression of alpha smooth muscle actin (alpha-SMA), collagen I, fibronectin, and laminin; and (iii) activation of MAPKs (extracellular regulated kinases 1 and 2, p38 kinase, and c Jun N terminal kinase). The effect of retinol on PSCs treated with ethanol was also examined by incubating cells with ethanol in the presence or absence of retinol for five days, followed by assessment of alpha-SMA, collagen I, fibronectin, and laminin expression. RESULTS: Retinol, ATRA, and 9-RA significantly inhibited: (i) cell proliferation, (ii) expression of alpha-SMA, collagen I, fibronectin, and laminin, and (iii) activation of all three classes of MAPKs. Furthermore, retinol prevented ethanol induced PSC activation, as indicated by inhibition of the ethanol induced increase in alpha-SMA, collagen I, fibronectin, and laminin expression. CONCLUSIONS: Retinol and its metabolites ATRA and 9-RA induce quiescence in culture activated PSCs associated with a significant decrease in the activation of all three classes of MAPKs in PSCs. Ethanol induced PSC activation is prevented by retinol supplementation. PMID- 16043493 TI - Transcript selection and the recruitment of mRNA decay factors for NMD in Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, nonsense-mediated mRNA decay (NMD) requires Upf1p, Upf2p, and Upf3p to accelerate the decay rate of two unique classes of transcripts: (1) nonsense mRNAs that arise through errors in gene expression, and (2) naturally occurring transcripts that lack coding errors but have built-in features that target them for accelerated decay (error-free mRNAs). NMD can trigger decay during any round of translation and can target Cbc-bound or eIF-4E bound transcripts. Extremely low concentrations of the Upf proteins relative to the total pool of transcripts make it difficult to understand how nonsense transcripts are selectively recruited. To stimulate debate, we propose two alternative mechanisms for selecting nonsense transcripts for NMD and for assembling components of the surveillance complex, one for the first (pioneer) round of translation, called "nuclear marking," and the other for subsequent rounds, called "reverse assembly." The model is designed to accommodate (1) the low abundance of NMD factors, (2) the role of nucleocytoplasmic shuttling proteins in NMD, (3) the independent and nonobligate order of assembly of two different subcomplexes of NMD factors, and (4) the ability of NMD to simultaneously reduce or eliminate the synthesis of truncated proteins produced by nonsense transcripts while down-regulating but not completely eliminating functional proteins produced from error-free NMD-sensitive transcripts PMID- 16043494 TI - Protein content of minimal and ancestral ribosome. AB - Minimal genome approaches seek to define the smallest gene complement compatible with modern-type cellular life on Earth. A consensus of computational and experimental approaches indicates that a minimal genome is close to 300 protein coding genes, if a rich medium is provided for cell growth. I relate ribosomal gene content in completely sequenced genomes to ribosomal subunit structure and approximate the protein components of the putative minimal ribosome and the ribosome of the Last Universal Common Ancestor of Life. Both sets contain between 35 and 40 proteins. There is evidence of protein-protein and protein-RNA displacement in the evolution of both ribosomal subunits. PMID- 16043495 TI - Evolvability of the mode of peptide binding by an RNA. AB - The HIV Rev-response element (RRE) RNA binds strongly to two unrelated peptides, the HIV Rev peptide and an RRE-binding aptamer, the RSG-1.2 peptide, at a similar site, but using distinct sets of interactions. In this study, the nucleotide base requirements for the binding of the RRE to the Rev and RSG-1.2 peptides were determined by selection of Rev- and RSG-1.2-binding RRE variants using a bacterial reporter system. As a result, distinct differences in the bases necessary for binding the two peptides were found in the upper stem of the RRE. Strikingly, single nucleotide changes in this region were found to switch the peptide-binding specificity of the RRE from a bifunctional Rev- and RSG-1.2 binding mode to either a Rev-specific or a RSG-1.2- specific mode, demonstrating how an RNA can evolve alternative binding strategies in discrete steps without intermediate loss of function. This evolvability of the mode of peptide binding by an RNA presumably reflects the multidimensionality of conformational space that a given RNA has available for ligand recognition, which may have been utilized in the evolution of RNA-polypeptide complexes. PMID- 16043496 TI - Lead(II) cleavage analysis of RNase P RNA in vivo. AB - The overall conformation of M1 RNA, the catalytic RNA subunit of RNase P in Escherichia coli, was analyzed in vivo and, in the presence of the C5 protein subunit, in vitro by lead(II) acetate probing. The partial cleavage patterns obtained are congruent with previous structure mapping performed in vitro. Most of the known major and minor cleavages in M1 RNA were supported and could be mapped onto a secondary structure model. The data obtained indicate that C5 has only minor effects on the overall structure of the RNA subunit. The similar cleavage patterns obtained in vitro and in vivo furthermore suggest that the intracellular environment does not greatly alter the overall conformation of M1 RNA within the holoenzyme complex. Moreover, our data indicate that M1 RNA in vivo is present in at least two states-the major fraction is bound to tRNA substrates and a minor fraction is substrate free. Finally, both in this and previous work we found that lead(II) probing data from in vivo experiments conducted on longer RNAs (tmRNA and M1 RNA) generally gives superior resolution compared to parallel in vitro experiments. This may reflect the absence of alternative conformers present in vitro and the more natural state of these RNAs in the cell due to proper, co-transcriptional folding pathways and possibly the presence of RNA chaperones. PMID- 16043497 TI - An optimized isolation and labeling platform for accurate microRNA expression profiling. AB - MicroRNAs (miRNAs) are small, noncoding RNAs that regulate gene expression in both plants and animals. miRNA genes have been implicated in a variety of important biological processes, including development, differentiation, apoptosis, fat metabolism, viral infection, and cancer. Similar to protein-coding messenger RNAs, miRNA expression varies between tissues and developmental states. To acquire a better understanding of global miRNA expression in tissues and cells, we have developed isolation, labeling, and array procedures to measure the relative abundance of all of the known human mature miRNAs. The method relies on rapid isolation of RNA species smaller than ~40 nucleotides (nt), direct and homogenous enzymatic labeling of the mature miRNAs with amine modified ribonucleotides, and hybridization to antisense DNA oligonucleotide probes. A thorough performance study showed that this miRNA microarray system can detect subfemtomole amounts of individual miRNAs from <1 mug of total RNA, with 98% correlation between independent replicates. The system has been applied to compare the global miRNA expression profiles in 26 different normal human tissues. This comprehensive analysis identified miRNAs that are preferentially expressed in one or a few related tissues and revealed that human adult tissues have unique miRNA profiles. This implicates miRNAs as important components of tissue development and differentiation. Taken together, these results emphasize the immense potential of microarrays for sensitive and high-throughput analysis of miRNA expression in normal and disease states. PMID- 16043498 TI - Specificity of recognition of mRNA 5' cap by human nuclear cap-binding complex. AB - The heterodimeric nuclear cap-binding complex (CBC) binds to the mono-methylated 5' cap of eukaryotic RNA polymerase II transcripts such as mRNA and U snRNA. The binding is important for nuclear maturation of mRNAs and possibly in the first round of translation and nonsense-mediated decay. It is also essential for nuclear export of U snRNAs in metazoans. We report characterization by fluorescence spectroscopy of the recognition of 5' capped RNA by human CBC. The association constants (K(as)) for 17 mono- and dinucleotide cap analogs as well as for the oligomer m7GpppA(m2') pU(m2')pA(m2') cover the range from 1.8 x 10(6) M(-1) to 2.3 x 10(8) M(-1). Higher affinity for CBC is observed for the dinucleotide compared with mononucleotide analogs, especially for those containing a purine nucleoside next to m7G. The mRNA tetramer associates with CBC as tightly as the dinucleotide analogs. Replacement of Tyr138 by alanine in the CBP20 subunit of CBC reduces the cap affinity except for the mononucleotide analogs, consistent with the crystallographic observation of the second base stacking on this residue. Our spectroscopic studies showed that contrary to the other known cap-binding proteins, the first two nucleotides of a capped-RNA are indispensable for its specific recognition by CBC. Differences in the cap binding of CBC compared with the eukaryotic translation initiation factor 4E (eIF4E) are analyzed and discussed regarding replacement of CBC by eIF4E. PMID- 16043499 TI - Activity-dependent polyadenylation in neurons. AB - Activity-dependent changes in protein synthesis modify synaptic efficacy. One mechanism that regulates mRNA translation in the synapto-dendritic compartment is cytoplasmic polyadenylation, a process controlled by CPEB, the cytoplasmic polyadenylation element (CPE)-specific RNA binding protein. In neurons, very few mRNAs are known CPEB substrates, and none appear to be responsible for the effects on plasticity that are found in the CPEB knockout mouse. These results suggest that the translation of other mRNAs is regulated by CPEB. To identify them, we have developed a functional assay based on the polyadenylation of brain derived mRNAs injected into Xenopus oocytes, a surrogate system that carries out this 3' end processing event in an efficient manner. The polyadenylated RNAs were isolated by binding to and thermal elution from poly(U) agarose and identified by microarray analysis. Selected sequences that were positive for polyadenylation were cloned and retested for polyadenylation by injection into oocytes. These sequences were then examined for activity-dependent polyadenylation in cultured hippocampal neurons. Finally, the levels of two proteins encoded by polyadenylated mRNAs were examined in glutamate-stimulated synaptoneurosomes. These studies show that many mRNAs undergo activity-dependent polyadenylation in neurons and that this process coincides with increased translation in the synapto dendritic compartment. PMID- 16043500 TI - A mutational analysis of U12-dependent splice site dinucleotides. AB - Introns spliced by the U12-dependent minor spliceosome are divided into two classes based on their splice site dinucleotides. The /AU-AC/ class accounts for about one-third of U12-dependent introns in humans, while the /GU-AG/ class accounts for the other two-thirds. We have investigated the in vivo and in vitro splicing phenotypes of mutations in these dinucleotide sequences. A 5' A residue can splice to any 3' residue, although C is preferred. A 5' G residue can splice to 3' G or U residues with a preference for G. Little or no splicing was observed to 3' A or C residues. A 5' U or C residue is highly deleterious for U12 dependent splicing, although some combinations, notably 5' U to 3' U produced detectable spliced products. The dependence of 3' splice site activity on the identity of the 5' residue provides evidence for communication between the first and last nucleotides of the intron. Most mutants in the second position of the 5' splice site and the next to last position of the 3' splice site were defective for splicing. Double mutants of these residues showed no evidence of communication between these nucleotides. Varying the distance between the branch site and the 3' splice site dinucleotide in the /GU-AG/ class showed that a somewhat larger range of distances was functional than for the /AU-AC/ class. The optimum branch site to 3' splice site distance of 11-12 nucleotides appears to be the same for both classes. PMID- 16043501 TI - Conformation of 4.5S RNA in the signal recognition particle and on the 30S ribosomal subunit. AB - The signal recognition particle (SRP) from Escherichia coli consists of 4.5S RNA and protein Ffh. It is essential for targeting ribosomes that are translating integral membrane proteins to the translocation pore in the plasma membrane. Independently of Ffh, 4.5S RNA also interacts with elongation factor G (EF-G) and the 30S ribosomal subunit. Here we use a cross-linking approach to probe the conformation of 4.5S RNA in SRP and in the complex with the 30S ribosomal subunit and to map the binding site. The UV-activatable cross-linker p-azidophenacyl bromide (AzP) was attached to positions 1, 21, and 54 of wild-type or modified 4.5S RNA. In SRP, cross-links to Ffh were formed from AzP in all three positions in 4.5S RNA, indicating a strongly bent conformation in which the 5' end (position 1) and the tetraloop region (including position 54) of the molecule are close to one another and to Ffh. In ribosomal complexes of 4.5S RNA, AzP in both positions 1 and 54 formed cross-links to the 30S ribosomal subunit, independently of the presence of Ffh. The major cross-linking target on the ribosome was protein S7; minor cross-links were formed to S2, S18, and S21. There were no cross-links from 4.5S RNA to the 50S subunit, where the primary binding site of SRP is located close to the peptide exit. The functional role of 4.5S RNA binding to the 30S subunit is unclear, as the RNA had no effect on translation or tRNA translocation on the ribosome. PMID- 16043502 TI - RNA secondary structure prediction by centroids in a Boltzmann weighted ensemble. AB - Prediction of RNA secondary structure by free energy minimization has been the standard for over two decades. Here we describe a novel method that forsakes this paradigm for predictions based on Boltzmann-weighted structure ensemble. We introduce the notion of a centroid structure as a representative for a set of structures and describe a procedure for its identification. In comparison with the minimum free energy (MFE) structure using diverse types of structural RNAs, the centroid of the ensemble makes 30.0% fewer prediction errors as measured by the positive predictive value (PPV) with marginally improved sensitivity. The Boltzmann ensemble can be separated into a small number (3.2 on average) of clusters. Among the centroids of these clusters, the "best cluster centroid" as determined by comparison to the known structure simultaneously improves PPV by 46.5% and sensitivity by 21.7%. For 58% of the studied sequences for which the MFE structure is outside the cluster containing the best centroid, the improvements by the best centroid are 62.5% for PPV and 31.4% for sensitivity. These results suggest that the energy well containing the MFE structure under the current incomplete energy model is often different from the one for the unavailable complete model that presumably contains the unique native structure. Centroids are available on the Sfold server at http://sfold.wadsworth.org. PMID- 16043503 TI - Molecular mimicry: quantitative methods to study structural similarity between protein and RNA. AB - With rapidly increasing availability of three-dimensional structures, one major challenge for the post-genome era is to infer the functions of biological molecules based on their structural similarity. While quantitative studies of structural similarity between the same type of biological molecules (e.g., protein vs. protein) have been carried out intensively, the comparable study of structural similarity between different types of biological molecules (e.g., protein vs. RNA) remains unexplored. Here we have developed a new bioinformatics approach to quantitatively study the structural similarity between two different types of biopolymers--proteins and RNA--based on the spatial distribution of conserved elements. We applied it to two previously proposed tRNA-protein mimicry pairs whose functional relatedness between two molecules has been recently determined experimentally. Our method detected the biologically meaningful signals, which are consistent with experimental evidence. PMID- 16043504 TI - Unorthodox mRNA start site to extend the highly structured leader of retrotransposon Tto1 mRNA increases transposition rate. AB - Retroelement RNAs serve as templates for both translation and reverse transcription into extrachromosomal DNA. DNA copies may be inserted into the host genome to multiply element sequences. This transpositional activity of retroelements is usually restricted to specific conditions, particularly to conditions that impose stress on the host organism. In this work, we examined how the mRNA initiation point, and features of primary and secondary structure, of tobacco retrotransposon Tto1 RNA influence its transpositional activity. We found that the most abundant Tto1 RNA is not a substrate for reverse transcription. It is poorly translated, and its 5'-end does not contain a region of redundancy with the most prominent 3'-end. In contrast, expression of an mRNA with the 5'-end extended by 28 nucleotides allows translation and gives rise to transposition events in the heterologous host, Arabidopsis thaliana. In addition, the presence of extended hairpins and of two short open reading frames in the 5'-leader sequence of Tto1 mRNA suggests that translation does not involve ribosome scanning from the mRNA 5'-end to the translation initiation site. PMID- 16043505 TI - FRET analyses of the U2AF complex localize the U2AF35/U2AF65 interaction in vivo and reveal a novel self-interaction of U2AF35. AB - We have analyzed the interaction between the U2AF subunits U2AF35 and U2AF65 in vivo using fluorescence resonance energy transfer (FRET) microscopy. U2 snRNP Auxiliary Factor (U2AF) is an essential pre-mRNA splicing factor complex, comprising 35-kDa (U2AF35) and 65-kDa (U2AF65) subunits. U2AF65 interacts directly with the polypyrimidine tract and promotes binding of U2 snRNP to the pre-mRNA branchpoint, while U2AF35 associates with the conserved AG dinucleotide at the 3' end of the intron and has multiple functions in the splicing process. Using two different approaches for measuring FRET, we have identified and spatially localized sites of direct interaction between U2AF35 and U2AF65 in vivo in live cell nuclei. While U2AF is thought to function as a heterodimeric complex, the FRET data have also revealed a novel U2AF35 self-interaction in vivo, which is confirmed in vitro using biochemical assays. These results suggest that the stoichiometry of the U2AF complex may, at least in part, differ in vivo from the expected heterodimeric complex. The data show that FRET studies offer a valuable approach for probing interactions between pre-mRNA splicing factors in vivo. PMID- 16043506 TI - Nop53p is required for late 60S ribosome subunit maturation and nuclear export in yeast. AB - We report that Ypl146cp/Nop53p is associated with pre-60S ribosomal complexes and localized to the nucleolus and nucleoplasm. In cells depleted of Nop53p synthesis of the rRNA components of the 60S ribosomal subunit is severely inhibited, with strikingly strong accumulation of the 7S pre-rRNA and a 5' extended form of the 25S rRNA. In cells depleted of Nop53p pre-60S subunits accumulate in the nucleus. However, a heterokaryon assay demonstrated that Nop53p is not transferred between nuclei, indicating that it is not released into the cytoplasm. We conclude that Nop53p is a late-acting factor in the nuclear maturation of 60S ribosomal subunits, which is required for normal acquisition of export competence. The strong accumulation of preribosomes in the Nop53p-depleted strain further suggests that it may participate in targeting aberrant preribosomes to surveillance and degradation pathways. PMID- 16043507 TI - Ribavirin is not a functional mimic of the 7-methyl guanosine mRNA cap. AB - Ribavirin is a guanosine ribonucleoside analog that displays broad-spectrum anti viral activity and is currently used for the treatment of some viral infections. Ribavirin has recently been proposed to also be a mimic of the 7-methyl guanosine cap found at the 5' end of mRNAs. To obtain supporting functional data for this hypothesis, we assessed the ability of ribavirin triphosphate to interfere with the interaction between eIF4E and 7-methyl guanosine capped mRNA. In chemical cross-linking assays, cap-affinity chromatography, and cap-dependent translation assays, ribavirin was unable to function as a cap analog. PMID- 16043508 TI - The bipartite structure of the tRNA m1A58 methyltransferase from S. cerevisiae is conserved in humans. AB - Among all types of RNA, tRNA is unique given that it possesses the largest assortment and abundance of modified nucleosides. The methylation at N(1) of adenosine 58 is a conserved modification, occurring in bacterial, archaeal, and eukaryotic tRNAs. In the yeast Saccharomyces cerevisiae, the tRNA 1 methyladenosine 58 (m(1)A58) methyltransferase (Mtase) is a two-subunit enzyme encoded by the essential genes TRM6 (GCD10) and TRM61 (GCD14). While the significance of many tRNA modifications is poorly understood, methylation of A58 is known to be critical for maintaining the stability of initiator tRNA(Met) in yeast. Furthermore, all retroviruses utilize m(1)A58-containing tRNAs to prime reverse transcription, and it has been shown that the presence of m(1)A58 in human tRNA(3) (Lys) is needed for accurate termination of plus-strand strong-stop DNA synthesis during HIV-1 replication. In this study we have identified the human homologs of the yeast m(1)A Mtase through amino acid sequence identity and complementation of trm6 and trm61 mutant phenotypes. When coexpressed in yeast, human Trm6p and Trm61p restored the formation of m(1)A in tRNA, modifying both yeast initiator tRNA(Met) and human tRNA(3) (Lys). Stable hTrm6p/hTrm61p complexes purified from yeast maintained tRNA m(1)A Mtase activity in vitro. The human m(1)A Mtase complex also exhibited substrate specificity--modifying wild type yeast tRNA(i) (Met) but not an A58U mutant. Therefore, the human tRNA m(1)A Mtase shares both functional and structural homology with the yeast tRNA m(1)A Mtase, possessing similar enzymatic activity as well as a conserved binary composition. PMID- 16043509 TI - Domain interactions within the Ski2/3/8 complex and between the Ski complex and Ski7p. AB - The Ski complex (composed of Ski3p, Ski8p, and the DEVH ATPase Ski2p) is a central component of the 3'-5' cytoplasmic mRNA degradation pathway in yeast. Although the proteins of the complex interact with each other as well as with Ski7p to mediate degradation by exosome, a 3'-exonuclease complex, the nature of these interactions is not well understood. Here we explore interactions within the Ski complex and between the Ski complex and Ski7p using a directed two-hybrid approach combined with coimmunoprecipitation experiments. We also test the functional significance of these interactions in vivo. Our results suggest that within the Ski complex, Ski3p serves as a scaffold protein with its C terminus interacting with Ski8p, and the sub-C terminus interacting with Ski2p, while no direct interaction between Ski2p and Ski8p was found. Ski7p interacts with the Ski complex via its interaction with Ski8p and Ski3p. In addition, inactivating the Ski complex by mutating conserved residues in the DEVH helicase motif of Ski2 did not abrogate its interaction with Ski7p, indicating that Ski2p function is not necessary for this interaction. PMID- 16043510 TI - The role of ribosome recycling factor in dissociation of 70S ribosomes into subunits. AB - Protein synthesis is initiated on ribosomal subunits. However, it is not known how 70S ribosomes are dissociated into small and large subunits. Here we show that 70S ribosomes, as well as the model post-termination complexes, are dissociated into stable subunits by cooperative action of three translation factors: ribosome recycling factor (RRF), elongation factor G (EF-G), and initiation factor 3 (IF3). The subunit dissociation is stable enough to be detected by conventional sucrose density gradient centrifugation (SDGC). GTP, but not nonhydrolyzable GTP analog, is essential in this process. We found that RRF and EF-G alone transiently dissociate 70S ribosomes. However, the transient dissociation cannot be detected by SDGC. IF3 stabilizes the dissociation by binding to the transiently formed 30S subunits, preventing re-association back to 70S ribosomes. The three-factor-dependent stable dissociation of ribosomes into subunits completes the ribosome cycle and the resulting subunits are ready for the next round of translation. PMID- 16043511 TI - Reversible intracellular translocation of KRas but not HRas in hippocampal neurons regulated by Ca2+/calmodulin. AB - The Ras/MAPK pathway regulates synaptic plasticity and cell survival in neurons of the central nervous system. Here, we show that KRas, but not HRas, acutely translocates from the plasma membrane (PM) to the Golgi complex and early/recycling endosomes in response to neuronal activity. Translocation is reversible and mediated by the polybasic-prenyl membrane targeting motif of KRas. We provide evidence that KRas translocation occurs through sequestration of the polybasic-prenyl motif by Ca2+/calmodulin (Ca2+/CaM) and subsequent release of KRas from the PM, in a process reminiscent of GDP dissociation inhibitor-mediated membrane recycling of Rab and Rho GTPases. KRas translocation was accompanied by partial intracellular redistribution of its activity. We conclude that the polybasic-prenyl motif acts as a Ca2+/CaM-regulated molecular switch that controls PM concentration of KRas and redistributes its activity to internal sites. Our data thus define a novel signaling mechanism that differentially regulates KRas and HRas localization and activity in neurons. PMID- 16043512 TI - Identification of FIP200 interaction with the TSC1-TSC2 complex and its role in regulation of cell size control. AB - FIP200 (focal adhesion kinase [FAK] family interacting protein of 200 kD) is a newly identified protein that binds to the kinase domain of FAK and inhibits its kinase activity and associated cellular functions. Here, we identify an interaction between FIP200 and the TSC1-TSC2 complex through FIP200 binding to TSC1. We found that association of FIP200 with the TSC1-TSC2 complex correlated with its ability to increase cell size and up-regulate S6 kinase phosphorylation but was not involved in the regulation of cell cycle progression. Conversely, knockdown of endogenous FIP200 by RNA interference reduced S6 kinase phosphorylation and cell size, which required TSC1 but was independent of FAK. Furthermore, overexpression of FIP200 reduced TSC1-TSC2 complex formation, although knockdown of endogenous FIP200 by RNA interference did not affect TSC1 TSC2 complex formation. Lastly, we showed that FIP200 is important in nutrient stimulation-induced, but not energy- or serum-induced, S6 kinase activation. Together, these results suggest a cellular function of FIP200 in the regulation of cell size by interaction with the TSC1-TSC2 complex. PMID- 16043513 TI - The Rho kinases I and II regulate different aspects of myosin II activity. AB - The homologous mammalian rho kinases (ROCK I and II) are assumed to be functionally redundant, based largely on kinase construct overexpression. As downstream effectors of Rho GTPases, their major substrates are myosin light chain and myosin phosphatase. Both kinases are implicated in microfilament bundle assembly and smooth muscle contractility. Here, analysis of fibroblast adhesion to fibronectin revealed that although ROCK II was more abundant, its activity was always lower than ROCK I. Specific reduction of ROCK I by siRNA resulted in loss of stress fibers and focal adhesions, despite persistent ROCK II and guanine triphosphate-bound RhoA. In contrast, the microfilament cytoskeleton was enhanced by ROCK II down-regulation. Phagocytic uptake of fibronectin-coated beads was strongly down-regulated in ROCK II-depleted cells but not those lacking ROCK I. These effects originated in part from distinct lipid-binding preferences of ROCK pleckstrin homology domains. ROCK II bound phosphatidylinositol 3,4,5P(3) and was sensitive to its levels, properties not shared by ROCK I. Therefore, endogenous ROCKs are distinctly regulated and in turn are involved with different myosin compartments. PMID- 16043514 TI - Mammalian WDR12 is a novel member of the Pes1-Bop1 complex and is required for ribosome biogenesis and cell proliferation. AB - Target genes of the protooncogene c-myc are implicated in cell cycle and growth control, yet the linkage of both is still unexplored. Here, we show that the products of the nucleolar target genes Pes1 and Bop1 form a stable complex with a novel member, WDR12 (PeBoW complex). Endogenous WDR12, a WD40 repeat protein, is crucial for processing of the 32S precursor ribosomal RNA (rRNA) and cell proliferation. Further, a conditionally expressed dominant-negative mutant of WDR12 also blocks rRNA processing and induces a reversible cell cycle arrest. Mutant WDR12 triggers accumulation of p53 in a p19ARF-independent manner in proliferating cells but not in quiescent cells. Interestingly, a potential homologous complex of Pes1-Bop1-WDR12 in yeast (Nop7p-Erb1p-Ytm1p) is involved in the control of ribosome biogenesis and S phase entry. In conclusion, the integrity of the PeBoW complex is required for ribosome biogenesis and cell proliferation in mammalian cells. PMID- 16043515 TI - The p85 regulatory subunit of phosphoinositide 3-kinase down-regulates IRS-1 signaling via the formation of a sequestration complex. AB - Phosphoinositide (PI) 3-kinase is required for most insulin and insulin-like growth factor (IGF) 1-dependent cellular responses. The p85 regulatory subunit of PI 3-kinase is required to mediate the insulin-dependent recruitment of PI 3 kinase to the plasma membrane, yet mice with reduced p85 expression have increased insulin sensitivity. To further understand the role of p85, we examined IGF-1-dependent translocation of p85alpha by using a green fluorescence protein (GFP)-tagged p85alpha (EGFP-p85alpha). In response to IGF-1, but not to PDGF signaling, EGFP-p85alpha translocates to discrete foci in the cell. These foci contain the insulin receptor substrate (IRS) 1 adaptor molecule, and their formation requires the binding of p85 to IRS-1. Surprisingly, monomeric p85 is preferentially localized to these foci compared with the p85-p110 dimer, and these foci are not sites of phosphatidylinositol-3,4,5-trisphosphate production. Ultrastructural analysis reveals that p85-IRS-1 foci are cytosolic protein complexes devoid of membrane. These results suggest a mechanism of signal down regulation of IRS-1 that is mediated by monomeric p85 through the formation of a sequestration complex between p85 and IRS-1. PMID- 16043516 TI - A local mechanism mediates NAD-dependent protection of axon degeneration. AB - Axon degeneration occurs frequently in neurodegenerative diseases and peripheral neuropathies. Important insight into the mechanisms of axon degeneration arose from findings that the degeneration of transected axons is delayed in Wallerian degeneration slow (Wlds) mice with the overexpression of a fusion protein with the nicotinamide adenine dinucleotide (NAD) synthetic enzyme, nicotinamide mononucleotide adenylyltransferase (Nmnat1). Although both Wld(s) and Nmnat1 themselves are functional in preventing axon degeneration in neuronal cultures, the underlying mechanism for Nmnat1- and NAD-mediated axon protection remains largely unclear. We demonstrate that NAD levels decrease in degenerating axons and that preventing this axonal NAD decline efficiently protects axons from degeneration. In support of a local protective mechanism, we show that the degeneration of axonal segments that have been separated from their soma could be prevented by the exogenous application of NAD or its precursor nicotinamide. Furthermore, we provide evidence that such Nmnat1/NAD-mediated protection is primarily mediated by their effects on local bioenergetics. Together, our results suggest a novel molecular pathway for axon degeneration. PMID- 16043517 TI - An essential role for c-FLIP in the efficient development of mature T lymphocytes. AB - Apoptosis-related genes play important roles in thymocyte maturation. We show that cellular FLICE-like inhibitory protein (c-FLIP), a procaspase-8-like apoptotic regulator, plays an essential role in the efficient development of mature T lymphocytes. Mice conditionally lacking c-FLIP in T lymphocytes display severe defects in the development of mature T cells, as indicated by a dramatically reduced number of CD4+ and CD8+ T cells in the spleen and lymph nodes of mutant mice. The impaired T lymphocyte maturation in c-FLIP conditional knockout mice occurs at the single-positive thymocyte stage and may be caused by enhanced apoptosis in vivo. Moreover, although c-FLIP has been implicated in T cell receptor signaling through nuclear factor (NF)-kappaB and Erk pathways, activation of NF-kappaB and Erk in c-FLIP-deficient thymocytes appears largely intact. Collectively, our data suggest that the primary role of c-FLIP in thymocyte maturation is to protect cells from apoptosis. PMID- 16043518 TI - Cellular FLICE-inhibitory protein is required for T cell survival and cycling. AB - Fas-associated death domain (FADD) and caspase-8 are key signal transducers for death receptor-induced apoptosis, whereas cellular FLICE-inhibitory protein (cFLIP) antagonizes this process. Interestingly, FADD and caspase-8 also play a role in T cell development and T cell receptor (TCR)-mediated proliferative responses. To investigate the underlying mechanism, we generated cFLIP-deficient T cells by reconstituting Rag-/- blastocysts with cFLIP-deficient embryonic stem cells. These Rag chimeric mutant mice (rcFLIP-/-) had severely reduced numbers of T cells in the thymus, lymph nodes, and spleen, although mature T lymphocytes did develop. Similar to FADD- or caspase-8-deficient cells, rcFLIP-/- T cells were impaired in proliferation in response to TCR stimulation. Further investigation revealed that cFLIP is required for T cell survival, as well as T cell cycling in response to TCR stimulation. Interestingly, some signaling pathways from the TCR complex appeared competent, as CD3 plus CD28 cross-linking was capable of activating the ERK pathway in rcFLIP-/- T cells. We demonstrate an essential role for cFLIP in T cell function. PMID- 16043519 TI - Tim-2 regulates T helper type 2 responses and autoimmunity. AB - Identification of the T cell immunoglobulin mucin-domain containing (Tim) gene family introduced a new family of cell surface molecules that is involved in the regulation of immune responses. We previously demonstrated that Tim-3 is expressed on terminally differentiated T helper (Th)1 cells, and serves to regulate Th1 immune responses. Here, we describe the identification and function of Tim-2, a novel member of the Tim gene family. In contrast with Tim-3, we demonstrate that Tim-2 is expressed preferentially in differentiated Th2 cells. Blockade of the Tim-2/Tim-2 ligand interaction, by administration of soluble Tim 2 fusion protein (Tim-2 immunoglobulin [Ig]), results in T cell hyperproliferation and the production of Th2 cytokines. Administration of Tim-2 Ig during the induction phase reduces the severity of experimental autoimmune encephalomyelitis, a Th1-mediated autoimmune disease model of multiple sclerosis. We propose that Tim-2, an orthologue of human Tim-1, is critical for the regulation of Th2 responses during autoimmune inflammation. PMID- 16043520 TI - Calcium-sensing soluble adenylyl cyclase mediates TNF signal transduction in human neutrophils. AB - Through chemical screening, we identified a pyrazolone that reversibly blocked the activation of phagocyte oxidase (phox) in human neutrophils in response to tumor necrosis factor (TNF) or formylated peptide. The pyrazolone spared activation of phox by phorbol ester or bacteria, bacterial killing, TNF-induced granule exocytosis and phox assembly, and endothelial transmigration. We traced the pyrazolone's mechanism of action to inhibition of TNF-induced intracellular Ca2+ elevations, and identified a nontransmembrane ("soluble") adenylyl cyclase (sAC) in neutrophils as a Ca2+-sensing source of cAMP. A sAC inhibitor mimicked the pyrazolone's effect on phox. Both compounds blocked TNF-induced activation of Rap1A, a phox-associated guanosine triphosphatase that is regulated by cAMP. Thus, TNF turns on phox through a Ca2+-triggered, sAC-dependent process that may involve activation of Rap1A. This pathway may offer opportunities to suppress oxidative damage during inflammation without blocking antimicrobial function. PMID- 16043521 TI - Multiple organ infection and the pathogenesis of SARS. AB - After >8,000 infections and >700 deaths worldwide, the pathogenesis of the new infectious disease, severe acute respiratory syndrome (SARS), remains poorly understood. We investigated 18 autopsies of patients who had suspected SARS; 8 cases were confirmed as SARS. We evaluated white blood cells from 22 confirmed SARS patients at various stages of the disease. T lymphocyte counts in 65 confirmed and 35 misdiagnosed SARS cases also were analyzed retrospectively. SARS viral particles and genomic sequence were detected in a large number of circulating lymphocytes, monocytes, and lymphoid tissues, as well as in the epithelial cells of the respiratory tract, the mucosa of the intestine, the epithelium of the renal distal tubules, the neurons of the brain, and macrophages in different organs. SARS virus seemed to be capable of infecting multiple cell types in several organs; immune cells and pulmonary epithelium were identified as the main sites of injury. A comprehensive theory of pathogenesis is proposed for SARS with immune and lung damage as key features. PMID- 16043522 TI - The challenge of providing doctors for rural America. PMID- 16043523 TI - The Appalachian Preceptorship: over two decades of an integrated clinical classroom experience of rural medicine and Appalachian culture. AB - There is a need to encourage careers in rural medicine and to prepare potential rural physicians for life in rural communities. The authors describe a program that addresses this need, the Appalachian Preceptorship Program, and report the program's experience from 1985 to 2004. The Appalachian Preceptorship is a four week summer elective conducted by the Department of Family Medicine of East Tennessee State University (ETSU) that offers students clinical preceptorships in rural areas of southern Appalachia. By the conclusion of the 2004 preceptorships, the program had served 225 medical students from 95 medical schools across the country and abroad. The program combines an individual community-based preceptorship with an interactive group instructional block, emphasizes rural medicine, and provides students an understanding of the interface between culture and medicine in southern Appalachia. Follow-up of Appalachian Preceptorship students during the 18-year period studied demonstrates that 82% of the 157 participants who matched before 2004 had selected residencies in primary care, with 60% entering family medicine. Those completing the program were more than three times as likely to practice in a rural community compared with the national average. Fifty-six percent of their practice settings carry multiple rural or underserved designations. The program has helped transform a legislative mandate to train doctors for rural communities into an institutional culture leading to more extensive programs and a greater recognition of ETSU's rural mission. The authors encourage other medical schools to develop combined clinical/classroom electives that reflect their institutional priorities and that can address a wide variety of clinical interests. PMID- 16043524 TI - Country doctors in literature: helping medical students understand what rural practice is all about. AB - Rural family medicine residencies and practices continue to have difficulty attracting applicants and practitioners. Students facing decisions about rural training or practice may be deterred by negative stereotypes or a lack of understanding about rural experience. Renewed efforts to foster students' interest and influence students' intent toward rural practice are sorely needed. The authors report one such innovative strategy that used literary sources, many written by rural physicians, to trigger discussion and reflection among a group of 11 medical students who volunteered in 2004 to participate in a two-day retreat sponsored by The Ohio State University College of Medicine Rural Health Scholars program. Participants first attended a presentation designed to help them understand the relevance of textual study of narratives by and about country doctors to their own experiences (during rural clerkships) in rural practice and as a vehicle for clarifying their concerns and questions. Through small-group study and discussion of excerpts from these texts, participants identified notable characteristics of rural inhabitants and their physicians; distinctive attitudes toward illness and medical care; and stresses and rewards of rural practice. They also wrote poems and essays in response to prompts about rural doctoring. Students used reading and writing as triggers to better comprehend and reflect on intangibles such as the nature of small-town life, relative professional isolation, and the unique aspects of the doctor-patient relationship in rural practice. Quantitative and qualitative evaluations suggest that this literature-based approach was enjoyable and stimulating for students, provided useful insights, and reinforced their interest in rural practice. PMID- 16043525 TI - Long-term retention of graduates from a program to increase the supply of rural family physicians. AB - PURPOSE: To determine the long-term retention of rural family physicians graduating from the Physician Shortage Area Program (PSAP) of Jefferson Medical College. METHOD: Of the 1,937 Jefferson graduates from the classes of 1978-1986, the authors identified those practicing rural family medicine when their practice location was first determined. The number and percent of PSAP and non-PSAP graduates practicing family medicine in the same rural area in 2002 were then identified, and compared to the number of those graduates practicing rural family medicine when they were first located in practice 11-16 years earlier. RESULTS: After 11-16 years, 68% (26/38) of the PSAP graduates were still practicing family medicine in the same rural area, compared with 46% (25/54) of their non-PSAP peers (p = .03). Survival analysis showed that PSAP graduates practice family medicine in the same rural locality longer than non-PSAP graduates (p = .04). CONCLUSIONS: These results are the first to show long-term rural primary care retention that is longer than the median duration. This outcome combined with previously published outcomes show that the PSAP represents the only program that has resulted in multifold increases in both recruitment (eight-fold) and long term retention (at least 11-16 years). In light of recent national recommendations to increase the total enrollment in medical schools, allocating some of this growth to developing and expanding programs similar to the PSAP would make a substantial and long lasting impact on the rural physician workforce. PMID- 16043526 TI - Current events: an important currency. PMID- 16043527 TI - An evaluation of the Rural Medical Education Program of the State University Of New York Upstate Medical University, 1990-2003. AB - PURPOSE: The Rural Medical Education Program (RMED) of the State University of New York (SUNY) Upstate Medical University is a 36-week clinical experience in rural communities for medical students that began in 1989. The authors sought to assess RMED's success in providing a valuable educational experience for students that assists rural communities recruit physicians. METHOD: In 2004, the authors used the Physician Masterfiles of the American Medical Association to compare practice locations of SUNY Upstate graduates who completed RMED with those who did not; surveyed former RMED students to assess their satisfaction with their practice location and the importance of RMED in helping them choose a location; interviewed hospital administrators in communities that have hosted RMED students to understand the impact of RMED on host communities; and compared United States Medical Licensing Examination Step 2 scores of RMED students with those of non RMED students to evaluate educational attainment. RESULTS: A greater percentage of former RMED students practiced in rural locations [22/86 (26%)] than did non RMED students [95/1,307 (7%)]. Ninety-one percent (69/76) of former RMED students were satisfied with their location, and 84% (64/76) believed that RMED was important in helping them choose a location. Hospital administrators viewed the program highly because it helped them recruit physicians and benefitted their medical staff. RMED students had higher adjusted mean Step 2 scores than did non RMED students (212.3 versus 199.1). CONCLUSION: The RMED program has successfully met its goals of providing a valuable educational experience for medical students and assisting rural communities recruit physicians. PMID- 16043528 TI - The impact on rural New Mexico of a family medicine residency. AB - PURPOSE: To determine the impact on rural New Mexico of the large, decentralized University of New Mexico (UNM) family medicine residency. METHOD: A cross sectional study was conducted of all 317 residency's graduates from 1974 to 2004. Location of current practice was correlated with the residents' gender, ethnicity, medical school of origin, and whether most training took place in the urban program or one of three rural programs. The residency's impact on rural communities was assessed. RESULTS: There was no significant gender difference between graduates who went into urban or rural practice. Compared with non minority graduates, a significantly greater percentage of ethnic minority graduates were in rural and urban New Mexico practices and fewer in out-of-state practices. A greater percentage of graduates who had been medical students in New Mexico practiced in both rural and urban New Mexico areas compared with graduates of out of state medical schools. Finally, a greater percentage of graduates from the three rural family medicine residencies remained in the state and practiced in rural areas compared with graduates from the urban program. The graduates' contributions to the school of medicine and to rural New Mexico are described. CONCLUSIONS: Graduates of UNM's family medicine residency have contributed significantly to the state's rural health workforce. Ethnic minority status, graduation from New Mexico's medical school, and training in one of the three rurally based residencies favored in-state and rural retention, while gender had no significant effect. The rural orientation of the residencies offered rural communities economic benefits. PMID- 16043529 TI - Viewpoint: exploring the human interior: the roles of cadaver dissection and radiologic imaging in teaching anatomy. AB - For a variety of reasons, new radiological imaging techniques are supplanting traditional cadaver dissection in the teaching of human anatomy. The authors briefly review the historical forces behind this transition, and then explore the advantages and drawbacks of each approach. Cadaver dissection offers an active, hands-on exploration of human structure, provides deep insights into the meaning of human embodiment and mortality, and represents a profound rite of passage into the medical profession. Radiological imaging permits in vivo visualization, offers physiologic as well as anatomic insights, and represents the context in which contemporary practicing physicians most frequently encounter their patients' otherwise hidden internal anatomy. Despite its important strengths, radiology cannot simply substitute for cadaver dissection, and the best models for teaching gross anatomy will incorporate both cadaver dissection and radiological imaging. PMID- 16043530 TI - A country doctor. PMID- 16043532 TI - Perceived impact of resident work hour limitations on medical student clerkships: a survey study. AB - PURPOSE: To assess medical students' perceptions of the impact of recent Accreditation Council for Graduate Medical Education policies limiting resident work hours on students' clerkship experiences, resident teaching, and quality of patient care. METHOD: In May/June 2003 and May/June 2004, an original questionnaire was administered to 252 medical students completing required clinical rotations at two teaching hospitals to assess students' perceptions of endpoints that might be affected by resident work hours limits. Response data were analyzed to determine statistical significance of differences between the two years studied. RESULTS: Questionnaires were completed by 129 students in 2003 (98%) and 112 students in 2004 (93%), for an overall response rate of 96%. A higher proportion of students perceived limits on work hours in 2004 [46 (41%)] than 2003 [36 (28%), p = .03]. Ratings of resident availability and primary resident's interest in teaching improved in 2004. Otherwise, ratings of the interest, skill, and availability of resident teachers and attending physicians remained stable between 2003 and 2004. Students reported spending similar amounts of time in formal teaching sessions and rated feedback similarly between 2003 and 2004. In 2004, fewer students [28 (25%)] reported considering leaving medicine due to long hours in training than in 2003 [49 (38%), p = .04]. No significant differences in the proportion of students reporting suboptimal care were found [44 (34%) in 2003, 34 (35%) in 2004, p = .57]. CONCLUSION: This small, early study suggests that reductions in resident work hours might be implemented without a significant negative impact upon medical students' self-assessed learning experiences, and that limiting resident work hours may even have a positive impact on medical students. PMID- 16043533 TI - The influence of the causes and contexts of medical errors on emergency medicine residents' responses to their errors: an exploration. AB - PURPOSE: To determine emergency medicine residents' emotional and behavioral responses to their medical errors and examine associations between residents' responses to medical error and perceptions of their training. METHOD: In 2003, 55 residents at two U.S. residency programs were asked to complete questionnaires about their errors and responses to their errors in three domains: emotional response, learning behavior, and disclosure. The questions were a mixture of free text, yes/no responses, and some were rated using a five-point Likert scale. Based on a conceptual framework, the authors constructed scales to describe the various domains and associations between the residents' responses to medical error and perceptions of their training were examined using Somers' D. RESULTS: A total of 43 residents returned questionnaires (80%); 40 of these residents described errors. Thirty-three (83%) residents discussed the error with someone; 27 (71%) with the attending and 10 (28%) with the patient/family. Negative emotions were common: 27 (68%) felt remorse, 21 (53%) guilt, 23 (58%) inadequacy, and 22 (55%) frustration. Residents' negative emotional responses were associated with their personal characteristics [26% (95% CI, 5-47%) association with lack of experience] and residents' perceptions of their training environment: 15% association with job overload (95% CI, -8-38%) and 23% association with lack of institutional support (95% CI, 5-41%). While 32 (81%) residents increased attention to detail, only 2 (5%) increased their use of evidence-based medicine. CONCLUSIONS: Errors committed by emergency medicine residents often resulted in negative emotions, limited constructive system-based improvements, and inadequate disclosure. Negative perceptions of the training environment are associated with negative emotional responses. PMID- 16043534 TI - The role of basic science knowledge and clinical knowledge in diagnostic reasoning: a structural equation modeling approach. AB - PURPOSE: To examine four theories on the role of basic science knowledge and clinical knowledge in diagnostic reasoning. METHOD: In 2000-01, the authors tested the basic science and clinical knowledge and diagnostic performances of 59 family physicians and 184 second- to sixth-year medical students at Maastricht University, The Netherlands. Structural equation modeling was used to analyze the data. Four theoretical models were tested. In the first model only basic science knowledge is involved in diagnostic reasoning; in the second model only clinical knowledge is related to diagnostic reasoning; in the third model, clinical knowledge is related to diagnostic reasoning, but basic science knowledge is integrated in clinical knowledge; and in the fourth model, both basic science knowledge and clinical knowledge independently influence diagnostic reasoning. RESULTS: Forty-four (75%) of the family physicians and 184 (100%) of the students responded. The results indicated that the third model, which is based on the knowledge encapsulation theory, provided the best fit to the data, whereas the models that had directly related basic science knowledge with diagnostic performance did not fit the data adequately. CONCLUSION: The results generally supported the third model by Schmidt and Boshuizen of knowledge encapsulation theory suggesting that basic science knowledge is activated in expert diagnostic reasoning through its relation with clinical knowledge. PMID- 16043535 TI - The relationship of insurance status, hospital ownership, and teaching status with interhospital transfers in California in 2000. AB - PURPOSE: Public hospitals and academic medical centers may admit more poorly insured transfer patients than do other institutions. The authors investigated the relationship of patient insurance status, hospital ownership, and hospital teaching status with interhospital transfers in California. METHOD: In 2003, data were derived from the hospital discharge abstract database for the year 2000 from the California Office of Statewide Health Planning and Development. Hospitals were categorized by ownership and teaching status; patients were categorized as being "good" or "poor" payers depending on the level of expected insurance reimbursement. Descriptive and multivariate analyses were used to assess the number of poor payer transfers admitted by each hospital group. RESULTS: In 2000, there were 58,509 transfer and 2,320,479 direct admissions. All hospital groups admitted a higher percentage of good payer than poor payer transfer patients (85% vs. 15% respectively for all groups combined). Adjusted for total number of admissions and teaching status, the number of poor payer transfer patients admitted to county-owned and University of California hospitals was significantly higher than the statewide average (both p values < .001), while the number admitted to independent teaching hospitals was significantly lower than the statewide average (p < .001). The number of poor payer transfer patients admitted to independent teaching hospitals more closely resembled that of for-profit hospitals than that of University of California teaching hospitals. CONCLUSIONS: In 2000, the likelihood of a hospital admitting a transfer patient appears to have been affected by both the patient's insurance status and the hospital's ownership. In general, good payer patients were more likely to be transferred than were poor payer patients, with poor payer transfer patients more likely to be admitted to publicly owned hospitals. PMID- 16043536 TI - Medical students' perceptions of medical education research and their roles as participants. AB - PURPOSE: To better understand whether medical students perceive medical education research as important to their medical training and whether published opinions about why medical students participate in research are accurate. METHOD: In 2003 04, 896 first- through fourth-year medical students at Kansas City University of Medicine and Biosciences College of Osteopathic Medicine were asked to complete an online eight-item questionnaire by responding Yes or No to each question. Responses were tallied by year of medical training and converted into numbers and percentages. Chi-square analysis was used to compare response rates among first- through fourth-year students and responses between preclinical and clinical students. RESULTS: A total of 524 students (58.5%) completed the questionnaire. A total of 488 (93%) medical students believed medical education research should be conducted to improve their medical training, 477 (91%) did not feel coerced to participate in studies because of faculty members' positions of authority, and 398 (76%) did not believe they would receive better grades, recommendations, and/or other favors. Four hundred sixty-eight (89%) students were not concerned with their confidentiality as study participants, while 326 (62%) wanted special protections. Response rates by year of medical school were not significantly different (p > .05). Responses of preclinical and clinical students for six of the eight questions were significantly different (p < .05). CONCLUSIONS: Medical school decisionmakers should recognize that students value medical education research. Published opinions about why medical students participate in studies are incongruent with medical students' views. Full review of medical education studies by Institutional Review Boards may be unnecessary and inappropriate. PMID- 16043537 TI - Disclosure of sexual orientation by medical students and residency applicants. AB - The authors report on sexual orientation disclosure issues considered by lesbian, gay, and bisexual students and physicians during their medical school and residency selection process. PMID- 16043539 TI - White privilege, color blindness, and services to battered women. AB - White privilege is a system of benefits, advantages, and opportunities experienced by White persons in our society simply because of their skin color. In this article, the authors present the results of a descriptive, exploratory study of White privilege in battered women's shelters in the Deep South. Based on a qualitative analysis, the authors show how White privilege is intricately connected to executive directors' claims of color blindness, the othering of women of color, and viewing White as the norm. The authors conclude the article with implications for service provision to battered women and directions for future research. PMID- 16043540 TI - Domestic violence at the intersections of race, class, and gender: challenges and contributions to understanding violence against marginalized women in diverse communities. AB - This article provides a comprehensive review of the emerging domestic violence literature using a race, class, gender, sexual orientation intersectional analysis and structural framework fostered by women of color and their allies to understand the experiences and contexts of domestic violence for marginalized women in U.S. society. The first half of the article lays out a series of challenges that an intersectional analysis grounded in a structural framework provides for understanding the role of culture in domestic violence. The second half of the article points to major contributions of such an approach to feminist methods and practices in working with battered women on the margins of society. PMID- 16043541 TI - The mediational role of relationship efficacy and resource utilization in the link between physical and psychological abuse and relationship termination. AB - This study examines the roles of physical and emotional abuse and resource utilization, relationship efficacy, and childhood abuse on relationship status (together or separated) in a sample of 69 low-income, nonsheltered battered women. Separate path models were conducted for physical and psychological abuse. Increased physical abuse was related to separated status, increased resource utilization, and decreased efficacy. The effect of physical abuse on status was mediated by resource utilization and efficacy, whereas the effect of psychological abuse on status was partially mediated only by utilization. Increased childhood abuse was associated with together status. Baseline psychological but not physical abuse predicted a longer term separated status thereby suggesting that the effects of psychological abuse may be enduring. PMID- 16043542 TI - Women's experience of violence during stalking by former romantic partners: factors predictive of stalking violence. AB - This study investigated female experiences of physical violence during stalking by a former romantic partner. It aimed to identify factors that were predictive of such stalking violence. Two hundred and twenty female undergraduates who defined themselves as victims of stalking following the dissolution of a romantic relationship completed a short questionnaire. From their responses, 11 predictor variables were considered. These were self-reported relationship experiences of physical and sexual violence, intentional damage to participant's property, partner jealousy, isolation, monitoring, criticism and insults by the former partner, former partner's drug and alcohol abuse, and specific threats of violence while being stalked. The dependent variable in the study was whether stalking violence occurred; 35.9% (79/220) of participants experienced stalking violence. Logistic regression analysis revealed that there were statistically significant independent associations between threats, partner jealousy, and former partner drug abuse and stalking violence. PMID- 16043543 TI - Adult domestic violence in cases of international parental child abduction. AB - This article discusses the Hague Convention on the Civil Aspects of International Child Abduction and its impact on battered mothers and their children seeking safety in the United States. We discuss relevant articles of the convention, the extent to which adult domestic violence is present in cases of international parental abduction, and cases in which battered mothers have contested the forced return of their children to an abusive partner. We conclude with recommended steps needed in research, training, and legislation that may increase the likelihood of safe outcomes for battered mothers and their children. PMID- 16043545 TI - Rape reporting after reforms: have times really changed? AB - Using data from the National Violence Against Women Survey, the authors examine whether rapes committed after reforms were more likely to be reported to police than those committed before reforms. The authors also consider whether the gap between the reporting of simple versus aggravated rape has narrowed. They find that rapes committed after 1990 were more likely to be reported than rapes occurring before 1974. Aggravated rape continues to be more likely to be reported than simple rape, however, and this effect is stable over time. The authors conclude by discussing the implications of these findings for evaluating the success of rape reform statutes. PMID- 16043546 TI - Attitudes toward rape: a comparison between Asian and Caucasian college students. AB - The purpose of this study is to investigate differences in attitudes toward rape between Asian and Caucasian college students. The Attitudes Toward Rape scale was used to measure beliefs about rape in a convenience sample of 169 college students. Three items regarding stranger rape myths were added. Findings suggest that Asian students are more likely than Caucasian students to believe women should be held responsible for preventing rape and to view sex as the primary motivation for rape. Asians also have stronger beliefs than Caucasians do that victims cause the rape and that most rapists are strangers. This research suggests that outreach programs can play an important role in providing information, education, and prevention regarding rape and that males and Asian students should be target populations for such programs. PMID- 16043547 TI - Regulating intimacy: judicial discourse in cases of wife assault (1970 to 2000). AB - This article describes the findings of research into judicial decision making in Ontario courts in cases of intimate violence against women. Judges are condemning the violence, issuing relatively harsh sentences, and arguing that the intimate context of the violence is an aggravating factor. The analysis also reveals that judges often rely on stereotypes and traditional notions of marriage, family, and femininity. As records of decisions, the documents suggest a high level of understanding that wife abuse is a crime. As judicial discourse, they reveal how the justice system regulates intimate relationships and how traditional ideologies persist despite the harsh sentences. PMID- 16043548 TI - TANF workers' responses to battered women and the impact of brief worker training: what survivors report. AB - Battered women (n = 159) report on their experiences with their Temporary Assistance to Needy Families (TANF) case workers. Workers most often ask about physical harm, feelings of fear, and police involvement. They least often create a safety plan, give information about work exemptions, and ask whether the partner had a gun. Women's major reasons for not talking about abuse are that the worker did not ask and a fear of negative outcomes. Workers who attended 1-day training are more likely than untrained workers to discuss the women's fear and physical harm, to help develop a safety plan, and to be viewed as generally helpful. PMID- 16043549 TI - Domestic violence: court case conviction and recidivism. AB - This article examines the effect of convictions for domestic violence or a related charge on the recidivism of batterers. Findings indicate that convictions reduce the likelihood of domestic violence recidivism. The variance explained by the conviction variable is modest; however, conviction remains significant when the batterers' histories of domestic violence charges and other background variables are introduced. The article also explores the associations between batterers' background characteristics and court convictions, as well as the effect of sanctions on the recidivism of convicted batterers. PMID- 16043551 TI - Barriers to domestic violence help seeking: implications for intervention. AB - Data were obtained from the Chicago Women's Health Risk Study, in which 491 abused women were interviewed in public health centers and a hospital. Responses of a subgroup of these women who did not seek help through the identified interventions are examined. Common themes emerge across the help-seeking interventions studied: individual thresholds for the seriousness of the violence, a perceived requirement to end the relationship, and certain specific barriers. The responses provide a glimpse into abused women's reasons for not seeking particular interventions. Implications for public health, mental health, domestic violence counseling agencies, and the police are discussed. PMID- 16043552 TI - Women's resources and use of strategies as risk and protective factors for reabuse over time. AB - Using a longitudinal and ecological approach, we investigated the relationships between women's material and emotional resources and strategies and their ability to stay safe over time in a sample of 406 help-seeking African American women. The multivariate analysis demonstrated that social support served as a protective factor and resistance strategies as risk factors for reabuse during a 1-year period. It also showed an interaction between social support and history of violence such that for participants who had experienced the most severe violence, social support did not serve as a protective factor; however, for the other participants, those with the least amount of social support had a 65% predicted probability of reabuse during the next year, compared to a 20% predicted probability for women reporting the highest level of social support. Policy and programmatic implications of these findings are discussed. PMID- 16043553 TI - Knowledge and attitudes about intimate partner violence among immigrant Latinos in rural North Carolina: baseline information and implications for outreach. AB - To create appropriate intimate partner violence (IPV) services for Latino immigrants, practitioners must be aware of their needs. We conducted interviews with 100 recent Latino immigrants in a rural North Carolina county. Overall, IPV was not perceived to be a problem; however, men and women differed in their perceptions. Men were more likely to agree with IPV myths, and both men and women felt that IPV had a detrimental impact on children. Many did not know about the local domestic violence agency, and knowledge about protective orders was limited. Outreach should emphasize the seriousness of IPV, adapt content for gender-specific audiences, and increase awareness about local resources. PMID- 16043554 TI - Individual, family, and community risk markers for domestic violence in Peru. AB - This study draws on an ecological framework in testing relationships between individual, family, and community characteristics and the likelihood of women experiencing domestic violence in Peru. The sample of 15,991 women was taken from the 2000 Peru Demographic and Health Survey. Logistic regression models revealed that at the individual level, low educational attainment, early union formation, and a violent family background increase a woman's likelihood of abuse. Family level risk markers include cohabitation, large family size, partner alcohol consumption, employment, and a woman's having higher status than her husband. At the community level, living in a noncoastal area and having an urban residence increase the likelihood of abuse. PMID- 16043555 TI - Understanding women's labeling of unwanted sexual experiences with dating partners: a qualitative analysis. AB - Qualitative data are used to examine the process by which women come to label their unwanted sexual experiences with dating partners as sexual abuse or assault, as well as the factors that impede such labeling. Issues of consent, causal attributions, and minimization were found to be central to understanding why women did or did not label. Labeling was generally a gradual process and was most often triggered by social support seeking. Women who did not label typically defined the incident as something less serious than sexual abuse or assault, although some provided definitions indicating that they had felt victimized. PMID- 16043557 TI - Revisiting unwanted sexual experiences on campus: a 12-year follow-up. AB - The current study examines patterns of stability and change in reports of unwanted sexual experiences in one campus community in two similar cohorts of undergraduates studied 12 years apart. A sample of 417 women completed a questionnaire in 2000, and this sample is compared to a sample of 524 women who completed the same questionnaire in 1988. Results indicate a reported decrease in unwanted sexual contact but indicate stability in reported rates of unwanted intercourse. Across forms of unwanted sexual experiences, more patterns of similarity than difference in abuse characteristics, such as relationship to perpetrator and location of the experience, are noted. PMID- 16043558 TI - Perceived causes of physical assault in heterosexual dating relationships. AB - Two studies investigated college students' perceptions of causes of violence in heterosexual dating relationships. Study 1 examined 107 participants' written causal explanations for dating violence. The second study focused on 70 students' ratings of cause in 15 scenarios ending with the man hitting the woman. Relationship and communication problems were primary causes when dating violence was presented in a concrete situation, whereas power and gender socialization were primary causes when presented as a social problem. Participants acknowledge power as an important cause, which suggests feminist structural frameworks in which relationship violence is embedded may lead to more effective prevention education. PMID- 16043559 TI - Sexual abuse of vulnerable young and old women: a comparative analysis of circumstances and outcomes. AB - Aggregated data from 125 substantiated Adult Protective Services cases of sexually abused women were collected during a 5-year period. Women older than 59 years represented 63% of the cases. Regardless of age, the most common types of abuse involved sexualized kissing and fondling and unwelcome sexual interest in the women's body. Most identified perpetrators were older males. Family members were most likely to abuse women living in the community, whereas women living in facilities usually experienced abuse by another resident. Perpetrators were prosecuted and convicted in six cases. About 12% of the women continued to be at risk of further sexual abuse. PMID- 16043560 TI - Trauma exposure and sexual revictimization risk: comparisons across single, multiple incident, and multiple perpetrator victimizations. AB - Although research demonstrates a link between child sexual abuse and sexual revictimization in adolescence or adulthood, less is known about specific mechanisms that increase women's vulnerability to reassault. This study examined experiential and outcome differences between survivors of a single assault, survivors of ongoing abuse by a single perpetrator, and survivors of multiple assaults by different offenders. Multiply victimized women differed from survivors of a single assault or of ongoing abuse on psychological distress, health, and nonsexual trauma variables. Revictimization by new perpetrators was predicted by an earlier age during a first sexual assault and by nonsexual trauma in childhood. PMID- 16043561 TI - What would MacGyver do? The meaning(s) of resistance and survival. AB - Drawing on interviews conducted with victims of New Zealand serial rapist, Malcolm Rewa, this article highlights the limitations of the terms resistance and survival as they are typically applied to women's responses to rape attacks. Although acknowledging that formulations that stress women's abilities to resist and their capacities to survive have been critically important in challenging popular notions of women as passive victims, the women's accounts presented here suggest a need to question whether such formulations are adequate to reflect the complexity and diversity of women's responses to sexual assaults. A preliminary attempt is also made to explore the implications associated with embracing expanded understandings of what resistance and survival might mean. PMID- 16043563 TI - Justice from the victim's perspective. AB - What are the meanings of justice, as seen from the perspective of victims of violent crime? Are victims' visions of justice represented by the conventional legal system? Are they represented by restorative justice? The author engages these questions, drawing on in-depth interviews with 22 victims of violent crime. It is argued that survivors' views of justice do not fit well into either retributive or restorative models. This has implications for current efforts to use restorative models in cases of violence against women. PMID- 16043564 TI - Gendered violence and restorative justice: the views of victim advocates. AB - The use of restorative justice for gendered violence has been debated in the feminist literature for some time. Critics warn that it is inappropriate because the process and outcomes are not sufficiently formal or stringent, and victims may be revictimized. Proponents assert that a restorative justice process may be better for victims than court because it holds offenders accountable and gives victims greater voice. This article presents what victim advocates in two Australian states think about using restorative justice for gendered violence. We find that although victim advocates have concerns and reservations about restorative justice, most saw positive elements. PMID- 16043565 TI - Sita's Trousseau: restorative justice, domestic violence, and South Asian culture. AB - This article focuses on the particular cultural factors that affect South Asian women who are abused and immigrant South Asian women who are abused, in particular, in the restorative justice process. By exploring cultural practices and the icon of Sita, the mythological heroine of the Ramayana, this article demonstrates how the South Asian ideals of womanhood and wifehood help to create a mind-set whereby South Asian women are reluctant to advocate for themselves and are reluctant to leave. Such a condition is contrary to the conditions and abilities assumed by the restorative justice movement for dispute resolution, inside or outside of domestic violence. It is concluded that restorative justice options are ill-suited to application among immigrant South Asian communities for domestic violence cases. PMID- 16043566 TI - Safety conferencing: toward a coordinated and inclusive response to safeguard women and children. AB - To reach out to women from different backgrounds, the battered women's movement needs to place women and their informal supports at the center of a coordinated response. This article shares the views of domestic violence survivors, staff, and supporters on how to create such a coordinated and inclusive response, lays a conceptual foundation for a decision-making forum called safety conferencing, and sets forth guidance for its practice. Safety conferencing is proposed as one means of building the individual and collective strength to reshape connections, make sound choices, and promote the safety of women and children from diverse cultures. PMID- 16043567 TI - Incorporating feminist theory and insights into a restorative justice response to sex offenses. AB - Sex offenses, particularly nonpenetration sex offenses and acquaintance sexual assault, are all too common. Because these crimes reinforce women's fear of crime and restrict spatial and social freedom, it is paramount for the justice system to act affirmatively; however, it does not. This article identifies several failures in the current response to these sex offenses. We describe the research demonstration project, RESTORE, operating in Pima County, Arizona, which uses a restorative justice response as a way of remedying some of those failures. Identifying central feminist insights that guided the development of that project, the article addresses concerns raised by feminists about the use of restorative justice for gendered violence. We conclude that most if not all of these concerns apply to cases of on going domestic violence--cases specifically excluded from the RESTORE program--rather than to cases of acquaintance sexual assault or nonpenetration sex offenses. PMID- 16043569 TI - The voices of battered women in India. AB - This study endeavors to give voice to marginalized, understudied women who were abused in India. The study included 90 women who were abused who were seeking help to address violence perpetrated against them by their husbands. A semistructured interview was used to collect data on abuse characteristics, consequences of abuse, and help-seeking behavior. Findings reveal that the vast majority of participants experienced daily abuse that was extremely severe, with 70% reporting they lived under the threat of murder. More than 85% reported needing medical care for their injuries. Respondents in this study sought help from a range of sources; the most effective help was provided by counseling centers and women's shelters. These findings have important implications for clinical practice and social policy. PMID- 16043570 TI - Do structural inequalities contribute to marital violence? Ethnographic evidence from rural South India. AB - Ethnographic research was conducted in rural communities in Karnataka State, South India, to explore the contexts in which marital violence occurs and the relationships between structural inequalities (gender, caste, and class inequalities) and marital violence. Research highlighted that (a) marital violence is intimately linked to experiences of gender, caste, and class inequalities; (b) women's ability to resist violence hinges on access to economic and social resources; and (c) health care providers need to be actively involved in responding to violence. This study demonstrates the urgent need for violence prevention initiatives, particularly those that address the contribution of structural inequalities. PMID- 16043571 TI - Challenging despair: teaching about women's resistance to violence. AB - In this article, the author describes an approach to teaching about violence against women that balances discussion of violence with information about women's individual and collective resistance. This strategy addresses two concerns about traditional approaches to this topic: that focusing only on victimization disempowers students and that it provides only a partial view of the reality of violence in women's lives. To address these problems, the author integrates discussion of resistance into the class's working definition of violence, assigned readings, guest speakers, and course assignments. The author concludes with a discussion of the positive effects of this approach. PMID- 16043572 TI - Training police officers on domestic violence and racism: challenges and strategies. AB - Teaching about domestic violence and racism is a formidable task that requires the inclusion of material that challenges dominant ideologies about race and gender. A recent training for police officers about domestic violence and racism serves as a good example of how challenging the task can be. Drawing from this 1 day police training, from existing pedagogical literature, and from the authors' collective experiences teaching about these issues, the authors address challenges encountered and propose strategies to mitigate them. The authors propose a multilevel approach to implementing strategies to address these challenges, which includes ongoing involvement during three stages of the training. PMID- 16043573 TI - When advocacy for domestic violence victims backfires. PMID- 16043575 TI - Walking a tightrope: the many faces of violence in the lives of racialized immigrant girls and young women. AB - This article explores a hidden yet pervasive form of violence that marks the lives of young women from racialized immigrant communities in western Canada. It argues for an intersectional analysis that takes into consideration their heightened vulnerability to systemic and institutional forms of violence. Situated at the intersections of race, class, gender, and age, these young women walk a tightrope between the violence of racism they experience from the host and/or dominant society and the pressures to conform imposed from within their communities. Challenging previous culturalist explanations, the article suggests that racism constitutes a significant form of structural violence experienced by these young women. PMID- 16043576 TI - Protective orders in rural and urban areas: a multiple perspective study. AB - This study is one of the first to examine the protective order process, barriers, and outcomes by combining qualitative and quantitative research in rural and urban areas. State statutes suggest the protective order process should be fair, consistent, and accessible to all women in the state. However, study results suggest that (a) the process of obtaining protective orders varies depending on community context; (b) although there are barriers to obtaining and enforcing protective orders regardless of geographic region, rural women appear to have more barriers; and (c) differences exist in victimization experiences, protective order stipulations, violations, and perceived effectiveness among rural and urban women. PMID- 16043577 TI - Restraining orders in California: a look at statewide data. AB - The authors tabulated statewide administrative data for all types of restraining orders. On June 6, 2003, there were 227,941 active restraining orders against adults in California; most were for domestic violence. Rates of restraining orders (i.e., restrained persons) were highest for men, African Americans, and 25 to 34-year-olds. In 72.2% of the orders, a woman was to be protected and a man was to be restrained; in 19.3%, the restrained and protected persons were of the same sex. Although state law prohibits the purchase or possession of a firearm by persons against whom a restraining order is issued, 9.2% of the orders documented no firearm restrictions. PMID- 16043578 TI - Flawed theory and method in studies of prostitution. PMID- 16043579 TI - Prostitution harms women even if indoors: reply to Weitzer. PMID- 16043580 TI - Reply to Weitzer. PMID- 16043583 TI - Children in the crossfire: child custody determinations among couples with a history of intimate partner violence. AB - Although most states mandate considerations of intimate partner violence (IPV) in child custody proceedings, little is known about how often a preexisting history of IPV is effectively presented to the courts in dissolution cases and, when it is, what effect it has on child custody and visitation outcomes. This retrospective cohort study examined the effects of a history of IPV, further categorized by whether substantiation of that history existed and whether the court handling the custody proceedings knew of that history, on child custody and visitation outcomes. The findings from this study highlight several issues of concern regarding the reality of child custody among families with a history of IPV. These include two primary concerns: a lack of identification of IPV even among cases with a documented, substantiated history, and a lack of strong protections being ordered even among cases in which a history of substantiated IPV is known to exist. PMID- 16043584 TI - Child custody mediation in cases of domestic violence: empirical evidence of a failure to protect. AB - Mediation of child custody disputes is mandated in several states. Investigators have averred that victims of domestic violence (DV) are greatly disadvantaged in mediation. The present study empirically evaluated outcomes and found that mediators failed to recognize and report DV in 56.9% of the DV cases. The court's screening form failed to indicate DV in at least 14.7% of the violent cases. Mediation resulted in poor outcomes for DV victims in terms of protections, such as supervised visitation and protected child exchanges. Mediator capacity to focus on the child's best interest was called into question. Child custody mediation should not be mandated in cases of DV. PMID- 16043585 TI - Outcomes of custody and visitation petitions when fathers are restrained by protection orders: the case of the New York family courts. AB - A random sample of custody and visitation petitions filed in New York City Family Courts in 1995 was used to examine outcomes of mothers' Order of Protection (OP) Petitions in relation to parents' custody and visitation petitions. Fathers restrained by OPs were more likely to secure visitation orders (64%) than not. In contrast, 80.8% of fathers' custody petitions were dismissed when they were restrained by OPs. Fathers' custody petitions were most likely to be ordered when mothers' OP petitions were withdrawn. Mothers were most likely to secure custody when their OP petitions were ordered or withdrawn. Courts rarely denied petitions. Those that did not result in court orders were either withdrawn by the petitioner or dismissed by the court (most likely because of failure of the petitioner to appear in court). This pattern has negative implications for battered women who may be vulnerable to pressure or threats from abusive ex partners. PMID- 16043586 TI - Child custody and visitation decisions when the father has perpetrated violence against the mother. AB - This research evaluated the effectiveness of statutes mandating a presumption against custody to a perpetrator of domestic violence (DV) and judicial education about DV. Across six states, the authors examined 393 custody and/or visitation orders where the father perpetrated DV against the mother and surveyed 60 judges who entered those orders. With the presumption, more orders gave legal and physical custody to the mother and imposed a structured schedule and restrictive conditions on fathers' visits, except where there was also a "friendly parent" provision and a presumption for joint custody. The presumption is effective only as part of a consistent statutory scheme. Although 86% of judges had received DV education, they scored no better in knowledge or attitudes. More of their orders gave mothers sole physical custody, and knowledge was associated with maternal custody, yet fewer structured or restricted fathers' visitation. Quality of DV education is more important than statutory mandate. PMID- 16043587 TI - Organic osmolytes as compatible, metabolic and counteracting cytoprotectants in high osmolarity and other stresses. AB - Organic osmolytes are small solutes used by cells of numerous water-stressed organisms and tissues to maintain cell volume. Similar compounds are accumulated by some organisms in anhydrobiotic, thermal and possibly pressure stresses. These solutes are amino acids and derivatives, polyols and sugars, methylamines, methylsulfonium compounds and urea. Except for urea, they are often called ;compatible solutes', a term indicating lack of perturbing effects on cellular macromolecules and implying interchangeability. However, these features may not always exist, for three reasons. First, some of these solutes may have unique protective metabolic roles, such as acting as antioxidants (e.g. polyols, taurine, hypotaurine), providing redox balance (e.g. glycerol) and detoxifying sulfide (hypotaurine in animals at hydrothermal vents and seeps). Second, some of these solutes stabilize macromolecules and counteract perturbants in non interchangeable ways. Methylamines [e.g. trimethylamine N-oxide (TMAO)] can enhance protein folding and ligand binding and counteract perturbations by urea (e.g. in elasmobranchs and mammalian kidney), inorganic ions, and hydrostatic pressure in deep-sea animals. Trehalose and proline in overwintering insects stabilize membranes at subzero temperatures. Trehalose in insects and yeast, and anionic polyols in microorganisms around hydrothermal vents, can protect proteins from denaturation by high temperatures. Third, stabilizing solutes appear to be used in nature only to counteract perturbants of macromolecules, perhaps because stabilization is detrimental in the absence of perturbation. Some of these solutes have applications in biotechnology, agriculture and medicine, including in vitro rescue of the misfolded protein of cystic fibrosis. However, caution is warranted if high levels cause overstabilization of proteins. PMID- 16043588 TI - A modified Hill muscle model that predicts muscle power output and efficiency during sinusoidal length changes. AB - The power output of a muscle and its efficiency vary widely under different activation conditions. This is partially due to the complex interaction between the contractile component of a muscle and the serial elasticity. We investigated the relationship between power output and efficiency of muscle by developing a model to predict the power output and efficiency of muscles under varying activation conditions during cyclical length changes. A comparison to experimental data from two different muscle types suggests that the model can effectively predict the time course of force and mechanical energetic output of muscle for a wide range of contraction conditions, particularly during activation of the muscle. With fixed activation properties, discrepancies in the work output between the model and the experimental results were greatest at the faster and slower cycle frequencies than that for which the model was optimised. Further optimisation of the activation properties across each individual cycle frequency examined demonstrated that a change in the relationship between the concentration of the activator (Ca2+) and the activation level could account for these discrepancies. The variation in activation properties with speed provides evidence for the phenomenon of shortening deactivation, whereby at higher speeds of contraction the muscle deactivates at a faster rate. The results of this study demonstrate that predictions about the mechanics and energetics of muscle are possible when sufficient information is known about the specific muscle. PMID- 16043589 TI - Effects of series elasticity and activation conditions on muscle power output and efficiency. AB - The activation of a muscle depends on the function that it is performing and on the architectural properties of that muscle; the two are inextricably linked. Activation conditions such as activation timing, duration and amplitude can be varied throughout a cyclical movement (such as locomotion) and the length change of the whole muscle tendon unit (MTU) can also be varied. Architecturally, muscles have a range of fibre lengths, maximum force-producing capabilities and stiffness of the series elastic element (SEE). In the present work we use a model to explore the relationship between power output and efficiency of a muscle across a range of contraction conditions. We have also examined the mechanical and energetic effects of changing muscle architecture within the model. Our results indicate that whilst there are clear optimal conditions for achieving maximum power output and maximum efficiency, the design of the muscle allows high levels of both to be achieved over a range of activation conditions. This range changes with both SEE compliance and the amplitude of the cyclical length change. The results suggest that a compliant SEE allows a muscle to function closer to the maximum of both power output and efficiency. In addition, by varying the amplitude of the activation level, the efficiency can theoretically remain unchanged, whilst the power output can be modulated. PMID- 16043590 TI - A year in the thermal life of a free-ranging herd of springbok Antidorcas marsupialis. AB - We used miniature data loggers implanted in the abdominal cavity to measure core body temperatures at 30 min intervals in eight (three males, five females) adult free-ranging springbok Antidorcas marsupialis in their natural habitat, over a period of 11-13 months. The animals were subjected to a nychthemeral range of air temperature that often exceeded 20 degrees C, with an absolute minimum temperature of -6 degrees C and a maximum of 34 degrees C. Abdominal temperature exhibited a low amplitude (approximately 1.2 degrees C) nychthemeral rhythm, with a temperature peak near sunset and a trough shortly after sunrise. The amplitude of the nychthemeral rhythm of body temperature was not correlated with the 24 h range of air temperature. Although mean 24 h body temperatures were positively correlated with corresponding air temperatures, mean daily body temperature increased, on average, by only 0.02 degrees C per 1 degrees C increase in air temperature, so that it was only approximately 0.3 degrees C higher in summer than in winter. Mean monthly body temperatures were strongly positively correlated with photoperiod and, in parallel with changes in the time of sunrise, the times at which the minimum and maximum body temperatures occurred were shifted approximately 1.2 h earlier in summer than in winter. Annual and daily variations in body temperature of springbok, like those of other free-living African ungulates, therefore appear to reflect an endogenous rhythm, entrained by the light:dark cycle, but largely independent of fluctuations in the environmental thermal load. Springbok exhibit remarkable homeothermy and do not employ adaptive heterothermy to survive in their natural environment. PMID- 16043591 TI - The olfactory pathway for individual recognition in the American lobster Homarus americanus. AB - Individual recognition in the lobster Homarus americanus (Milne-Edwards), is based on detection of urine pheromones via chemoreceptors of the lateral antennular flagellum. The specific sensory pathway mediating this recognition is not known. Most of the chemoreceptor cells of this flagellum are found in the unimodal aesthetasc sensilla and project specifically to the glomeruli of the olfactory lobe in the brain. Additional chemoreceptor cells are located among mechanoreceptor cells in bimodal sensilla, including the guard hairs; they do not project to the olfactory lobe. This neuroanatomy suggested that aesthetascs were essential to all complex chemosensory tasks until it was shown that spiny lobsters Panulirus argus can still perform complex food odor discrimination and localization tasks without aesthetascs. Here, we demonstrate that the aesthetascs of H. americanus contain the chemoreceptors necessary for individual recognition of familiar opponents. In contrast to intact and guard hair-shaved animals, lobsters with aesthetascs removed did not recognize previous opponents as shown by second encounters statistically similar in length and aggression to first encounter fights. Non-aesthetasc chemosensory pathways were incapable of rescuing opponent recognition. Subsequent lesion of all remaining chemoreceptor cells (by immersion in distilled water) abolished recognition and renewed fighting. PMID- 16043592 TI - Intraspecific variation of venom injected by fish-hunting Conus snails. AB - Venom peptides from two species of fish-hunting cone snails (Conus striatus and Conus catus) were characterized using microbore liquid chromatography coupled with matrix-assisted laser desorption/ionization-time of flight-mass spectrometry and electrospray ionization-ion trap-mass spectrometry. Both crude venom isolated from the venom duct and injected venom obtained by milking were studied. Based on analysis of injected venom samples from individual snails, significant intraspecific variation (i.e. between individuals) in the peptide complement is observed. The mixture of peptides in injected venom is simpler than that in the crude duct venom from the same snail, and the composition of crude venom is more consistent from snail to snail. While there is animal-to-animal variation in the peptides present in the injected venom, the composition of any individual's injected venom remains relatively constant over time in captivity. Most of the Conus striatus individuals tested injected predominantly a combination of two neuroexcitatory peptides (s4a and s4b), while a few individuals had unique injected-venom profiles consisting of a combination of peptides, including several previously characterized from the venom duct of this species. Seven novel peptides were also putatively identified based on matches of their empirically derived masses to those predicted by published cDNA sequences. Profiling injected venom of Conus catus individuals using matrix-assisted laser desorption/ionization-time of flight-mass spectrometry demonstrates that intraspecific variation in the mixture of peptides extends to other species of piscivorous cone snails. The results of this study imply that novel regulatory mechanisms exist to select specific venom peptides for injection into prey. PMID- 16043593 TI - A concept of dietary dipeptides: a step to resolve the problem of amino acid availability in the early life of vertebrates. AB - The premise that a dietary dipeptide approach will improve the understanding of amino acid utilization in the fastest-growing vertebrate, the teleost fish, was tested by examining the muscle free amino acid (FAA) pool and enzyme activities, in concert with growth response, when dietary amino acids were provided in free, dipeptide or protein molecular forms. We present the first evidence in fish that, in response to a synthetic dipeptide diet, muscle FAA varies as a result of both growth rate and amino acid availability of specific peptides. We demonstrate significantly diminished muscle indispensable FAA (3-10-fold) in rainbow trout alevins fed a dipeptide-based diet compared with a protein-based diet. The dipeptide-based diet did not contain proline, resulting in 10-27-fold less muscle free proline and hydroxyproline in alevins. The response of alevins fed FAA-based or peptide-based diets can be indicative of collagen turnover (Hyp/Pro ratio) and showed significant differences between dietary treatments. Pyrroline-5 carboxylate (P5C) reductase activity was detected, suggesting that P5C may ameliorate proline deficiency, but synthesis from glutamate could not maintain free proline levels in muscle. This finding will provide an impetus to test whether proline is conditionally indispensable in young fish, as in mammals and birds. This study shows that amino acids given entirely as dipeptides can sustain fish growth, result in muscle FAA and enzyme responses in line with dietary levels and identify growth-limiting amino acids. The understanding of these factors necessitates a diet formulation that will improve the accuracy of determining amino acid requirements in the early life stages of vertebrates. PMID- 16043594 TI - Contractile activity of the pectoralis in the zebra finch according to mode and velocity of flap-bounding flight. AB - We studied flying zebra finch (Taeniopygia guttata, N = 12), to provide a new test of a long-standing ;fixed-gear' hypothesis that flap-bounding birds use only intermittent non-flapping phases, instead of variation in muscle activity, to vary mechanical power output in flight. Using sonomicrometry and electromyography, we measured in vivo fascicle length and neuromuscular recruitment in the pectoralis as the birds flew in different flight modes (level, ascending, descending; mean velocity 1.6+/-0.3 m s(-1)) and across velocities in a new, variable-speed wind tunnel (0-12 m s(-1)). Synchronized high-speed digital video (250 Hz) provided a record of wing kinematics. Flight mode had a significant effect upon pectoralis strain, strain rate, fractional shortening and the relative timing of muscle activity (onset, offset and duration). Among flight velocities, we observed significant variation in pectoralis strain, fractional lengthening and shortening, strain rate, relative electromyographic (EMG) amplitude, and EMG duration and offset. In particular, variation in strain rate and relative EMG amplitude indicates that the fixed-gear hypothesis should be rejected. Instead, it appears that zebra finch vary work and power output within wingbeats by modulating muscle contractile behavior and between wingbeats using intermittent bounds. Muscle activity patterns and wing kinematics were similar between free flight and wind tunnel flight at similar speeds. Comparing flights with and without surgically implanted transducers and electrodes, zebra finch exhibited a reduction in maximum velocity (from 14 to 12 m s(-1)) and a significant increase in wingbeat frequency and percent time flapping. This identifies a potential limitation of in vivo flight measurements, and similar studies of bird flight should, therefore, include measurements of the extent to which flight performance is compromised by experimental protocol. PMID- 16043595 TI - Temporal organization of bi-directional traffic in the ant Lasius niger (L.). AB - Foraging in ants is generally organized along well-defined trails supporting a bi directional flow of outbound and nestbound individuals and one can hypothesize that this flow is maximized to ensure a high rate of food return to the nest. In this paper we examine the effect of bottlenecks on the temporal organization of ant flow. In our experiments ants had to cross a bridge to go from their nest to a food source. Two types of bridges were used: one with and one without bottlenecks. Traffic counts show that, in spite of the bottlenecks and the reduction of path width, the volume of traffic and the rate of food return were the same on both bridges. This was due to a change in the temporal organization of the flow: when path width decreases alternating clusters of inbound and outbound ants were observed crossing the bridge. This organization limits the number of head-on encounters and thus allows to maintain the same travel duration as on the wide bridge. A model is proposed to assess in various conditions the importance of the behavioural rules observed at the individual level for the regulation of traffic flow. It highlights how the interplay between the value of the flow and cooperative behaviours governs the formation and size of the clusters observed on the bridge. PMID- 16043596 TI - Mechanical properties of the integument of the common gartersnake, Thamnophis sirtalis (Serpentes: Colubridae). AB - The evolution of the ophidian feeding mechanism has involved substantial morphological restructuring associated with the ability to ingest relatively large prey. Previous studies examining the morphological consequences of macrophagy have concentrated on modifications of the skull and cephalic musculature. Although it is evident that macrophagy requires highly compliant skin, the mechanical properties of the ophidian integument have received limited attention, particularly in the context of feeding. We examined mechanical properties of skin along the body axis in Thamnophis sirtalis (Colubridae). Data were collected from tensile tests and were analyzed using a multivariate analysis of variance (MANOVA) and post-hoc multiple comparison tests. Significant differences in mechanical properties were detected among regions of the body. In general, prepyloric skin is more compliant than postpyloric skin, consistent with the demands of macrophagy. PMID- 16043597 TI - Metabolic responses to cold in subterranean crustaceans. AB - Changes in polyol, sugar and free amino acid (FAA) body contents were investigated in the aquatic, subterranean (i.e. hypogean) crustaceans Niphargus rhenorhodanensis and Niphargus virei and in a morphologically close aquatic, surface-dwelling (i.e. epigean) crustacean Gammarus fossarum acclimated to 12 degrees C, 3 degrees C and -2 degrees C. With decreasing temperature, G. fossarum significantly increased its alanine and glutamine levels, while trehalose body content was found to increase above control levels only at -2 degrees C. N. virei showed moderate increases of alanine and glycine, and no change in trehalose level was observed in this species. By contrast, N. rhenorhodanensis was the only species showing a significant rise in its total FAA pool, mainly explained by alanine, glycine, arginine and glutamine accumulations. This species also gradually increased its trehalose body content with decreasing temperature. Several cold-hardy ectotherms show metabolic responses to cold that are identical to those observed in N. rhenorhodanensis. A previous comparative study showed that the hypogean N. rhenorhodanensis exhibited a survival time (Lt50) at -2 degrees C that was 26.3 times and 2.6 times higher than the hypogean N. virei and the epigean G. fossarum, respectively. Thus, crustacean levels of FAA and trehalose were correlated with their respective cold tolerances. Such differences in metabolic responses to cold in both hypogean organisms were unexpected since they both live in thermally buffered biotopes. Considering the current distribution areas of the two subterranean crustaceans studied, we assume that the cold hardiness found in the hypogean N. rhenorhodanensis could be correlated with its biogeography history during the quaternary glaciations. PMID- 16043598 TI - Flow and conduit formation in the external fluid-transport system of a suspension feeder. AB - To what extent is the development of a fluid-transport system related to flow within the system? Colonies of the bryozoan Membranipora membranacea have a simple external fluid-transport system with three components: the canopy of lophophores (crowns of ciliated tentacles), the edge of the canopy, and chimneys (raised openings in the canopy). The lophophores pump seawater into the colony and capture food particles from the seawater. The chimneys and canopy edge let the water back out of the colony. New chimneys form at the canopy edge as the colony grows. I tested whether there was a correlation between chimney formation and excurrent flow speed at the canopy edge by measuring excurrent flow speeds prior to chimney formation. Excurrent flow speeds were higher in regions that produced chimneys than in regions that did not form chimneys. Observations of changes in chimney shape after anesthetization with MgCl2 suggest that both growth and behavior determine chimney shape. Together, the results suggest that there is a strong correlation between growth and flow in this external fluid transport system, with new chimneys forming at sites of high flow. PMID- 16043599 TI - Aquatic wing flapping at low Reynolds numbers: swimming kinematics of the Antarctic pteropod, Clione antarctica. AB - We studied swimming kinematics of the Antarctic pteropod, Clione antarctica, to investigate how propulsive forces are generated by flexible oscillating appendages operating at low Reynolds numbers (1025) exhibited gliding during the recovery phase of each half stroke. Maximum translational and rotational accelerations of the body occurred at the initiation of each power phase, suggesting that rotational circulation, the acceleration reaction, and wake recapture may all potentially contribute to vertical force production. Individual contributions of these mechanisms cannot, however, be assessed from these kinematic data alone. During recovery phases of each half-stroke, C. antarctica minimized adverse drag forces by orienting the wings parallel to flow and by moving them along the body surface, possibly taking advantage of boundary layer effects. Vertical force production was altered through changes in the hydrodynamic angle of attack of the wing that augmented drag during the power phase of each half-stroke. At higher translational velocities of the body, the inclination of the power phase also became more nearly vertical. These results indicate that, in addition to serotonin-mediated modulation of wingbeat frequency reported previously in Clione, geometric alteration of wingbeat kinematics offers a precise means of controlling swimming forces. PMID- 16043600 TI - Nitric oxide in control of luminescence from hatchetfish (Argyropelecus hemigymnus) photophores. AB - Nitric oxide synthase-like immunoreactivity (NOS-LI IR) was detected by immunohistochemistry in ventral light organs of the mesopelagic fish, Argyropelecus hemigymnus. Strong NOS-LI IR was present in nerve fibres and in other cells central for production or modulation of light: immunoreactive fibres surrounded the photophores, and were also present in the filter area. Filter cells, particularly in the outer layers, showed strong IR throughout the cytoplasm. Pharmacological studies suggested that nitric oxide (NO) modulates adrenaline-stimulated light emission, and that the modulation is correlated to the ability of the light organ to respond to adrenaline. Adrenaline is known to produce two different types of light response in isolated photophores from Argyropelecus: a slow, long-lasting, high intensity response, or a fast and weak response of short duration. Incubation of photophores in the NO donors sodium nitroprusside or S-nitroso-N-acetylpenicillamine prior to adrenaline stimulation reduced the intensity of the strong and long-lasting type of response, but had little or even a potentiating effect on the weakly responding photophores. Hydroxylamine, which is converted to NO if catalase activity is present in the tissue, reduced the duration and the intensity of the adrenaline response in all tested organs. The NOS-inhibitor L-thiocitrulline potentiated the adrenaline response in the weakly responding organs; the weaker the adrenaline effect, the stronger the potentiation caused by L-thiocitrulline. The strongly responding organs were instead inhibited by L-thiocitrulline. The results suggest that NO has an important role in the control of light emission from Argyropelecus hemigymnus photophores. The cGMP analogue dibutyryl cGMP, the guanylate cyclase inhibitor ODQ and the phosphodiesterase inhibitor pentoxiphylline had no effect, indicating that the NO effect does not involve cGMP. PMID- 16043601 TI - Carbon dioxide instantly sensitizes female yellow fever mosquitoes to human skin odours. AB - Female mosquitoes are noted for their ability to use odours to locate a host for a blood meal. Two sensory organs contribute to their sense of smell: the maxillary palps, which measure the level of CO2, and the antennae, which detect other host-released odours. To establish the relative importance and interactions of CO2 and other body emissions in freely flying mosquitoes, we presented female yellow fever mosquitoes Aedes aegypti L. with broad plumes of human skin odour and CO2 at natural concentrations and dilutions thereof in a wind tunnel. 3-D video-recorded flight tracks were reconstructed. Activation, flight velocity, upwind turning and source finding waned quickly as skin odours were diluted, whereas in the presence of CO2 these parameters remained unchanged over more than a 100-fold dilution from exhaled concentrations. Although mosquitoes were behaviourally less sensitive to skin odours than to CO2, their sensitivity to skin odours increased transiently by at least fivefold immediately following a brief encounter with a filament of CO2. This sensitization was reflected in flight velocity, track angle, turning rate upon entering and exiting the broad odour plume and, ultimately, in the source-finding rate. In Ae. aegypti, CO2 thus functions as a ;releaser' for a higher sensitivity and responsiveness to skin odours. The initially low responsiveness of mosquitoes to skin odours, their high sensitivity to CO2, and the sensitization of the olfactory circuitry by CO2 are ecologically relevant, because rapidly fluctuating CO2 levels reliably signal a potential host. Possible mechanisms of the instantaneous sensitization are considered. PMID- 16043602 TI - Air sac PO2 and oxygen depletion during dives of emperor penguins. AB - In order to determine the rate and magnitude of respiratory O2 depletion during dives of emperor penguins (Aptenodytes forsteri), air sac O2 partial pressure (PO2) was recorded in 73 dives of four birds at an isolated dive hole. These results were evaluated with respect to hypoxic tolerance, the aerobic dive limit (ADL; dive duration beyond which there is post-dive lactate accumulation) and previously measured field metabolic rates (FMRs). 55% of dives were greater in duration than the previously measured 5.6-min ADL. PO2 and depth profiles revealed compression hyperoxia and gradual O2 depletion during dives. 42% of final PO2s during the dives (recorded during the last 15 s of ascent) were <20 mmHg (<2.7 kPa). Assuming that the measured air sac PO2 is representative of the entire respiratory system, this implies remarkable hypoxic tolerance in emperors. In dives of durations greater than the ADL, the calculated end-of-dive air sac O2 fraction was <4%. The respiratory O2 store depletion rate of an entire dive, based on the change in O2 fraction during a dive and previously measured diving respiratory volume, ranged from 1 to 5 ml O2 kg(-1) min(-1) and decreased exponentially with diving duration. The mean value, 2.1+/-0.8 ml O2 kg(-1) min( 1), was (1) 19-42% of previously measured respiratory O(2) depletion rates during forced submersions and simulated dives, (2) approximately one-third of the predicted total body resting metabolic rate and (3) approximately 10% of the measured FMR. These findings are consistent with a low total body metabolic rate during the dive. PMID- 16043603 TI - Optical studies of nicotinic acetylcholine receptor subtypes in the guinea-pig enteric nervous system. AB - Nicotinic transmission in the enteric nervous system (ENS) is extensive, but the role of individual nicotinic acetylcholine receptor (nAChR) subtypes in the functional connectivity of its plexuses has been elusive. Using monoclonal antibodies (mAbs) against neuronal alpha3-, alpha4-, alpha3/alpha5-, beta2-, beta4- and alpha7-subunits, combined with radioimmunoassays and immunocytochemistry, we demonstrate that guinea-pig enteric ganglia contain all of these nAChR-subunits with the exception of alpha4, and so, differ from mammalian brain. This information alone, however, is insufficient to establish the functional role of the identified nAChR-subtypes within the enteric networks and, ultimately, their specific contributions to gastrointestinal physiology. We have used voltage-sensitive dyes and a high-speed CCD camera, in conjunction with specific antagonists to various nAChRs, to elucidate some of the distinct contributions of the individual subtypes to the behaviour of enteric networks. In the guinea-pig, the submucous plexus has the extraordinary advantage that it is virtually two-dimensional, permitting optical recording, with single cell resolution, of the electrical activity of all of its neurones. In this plexus, the block of alpha3beta2-, alpha3beta4- and/or alpha7-nAChRs always results in a decrease in the magnitude of the synaptic response. However, the magnitude of the fast excitatory post-synaptic potentials (epsps) evoked by electrical stimulation of a neighbouring ganglion varies from cell to cell, reflecting the differential expression of subunits already observed using mAbs, as well as the strengths of the activated synaptic inputs. At the same time, we observe that submucous neurones have a substantial mecamylamine (Mec)-insensitive (non-nicotinic) component to their fast epsps, which may point to the presence of purinergic or serotonergic fast epsps in this system. In the myenteric plexus, on the other hand, the antagonist-induced changes in the evoked synaptic response vary depending upon the location of the stimulating electrode with respect to the ganglion under study. The range of activity patterns that follows sequential pharmacological elimination of individual subtypes suggests that nAChRs may be capable of regulating the activity of both excitatory and inhibitory pathways, in a manner similar to that described in the central nervous system. PMID- 16043604 TI - Ontogenetic effects on aerobic and anaerobic metabolism during jumping in the American locust, Schistocerca americana. AB - Developing vertebrates increase both their locomotory power output and endurance due to ontogenetic improvements in anaerobic and aerobic metabolic capacities. Do similar patterns hold for insect locomotion, or do longer tracheal lengths create problems for oxygen delivery in older animals? We forced developing American locust grasshoppers (Schistocerca americana) to jump repeatedly and examined the effect of development on power output, endurance, lactate concentration, oxygen consumption and the oxygen sensitivity of jump performance. As previously shown, power outputs, relative leg lengths and leg cuticular content increased with age. A key finding of this study is that both lactate concentration and aerobic metabolic rate of the jumping muscle increase with age, explaining how the increased leg cuticular stiffness can result in increased power output. After two minutes of jumping, grasshoppers rely completely on aerobic ATP production. The rise in mass-specific, active aerobic metabolic rates with age indicates that problems with longer tracheae can be overcome; however, the reduced endurance, higher lactate concentrations and increased oxygen sensitivity of locomotory performance in older animals indicate that larger/older grasshoppers have smaller safety margins for oxygen delivery during hopping. PMID- 16043605 TI - Rethinking evidence-based practice for children's mental health. PMID- 16043607 TI - Functional impairment recovers after episodes of major depression. PMID- 16043608 TI - Review: depression may increase mortality in coronary heart disease. PMID- 16043609 TI - Review: after myocardial infarction, depression and poor prognosis are associated. PMID- 16043610 TI - People diagnosed with schizophrenia retain the diagnosis long term. PMID- 16043611 TI - The Screening Tool for Autism in Two Year Olds can identify children at risk of autism. PMID- 16043612 TI - Middelheim Frontality Score may be useful for differentiating between Alzheimer's disease and frontotemporal dementia. PMID- 16043613 TI - Review: hypericum extracts are safer and lead to fewer adverse effects than older standard antidepressants, but have similar tolerability to SSRIs. PMID- 16043614 TI - Metyrapone is an effective adjuvant treatment for major depression. PMID- 16043615 TI - Bright light treatment an effective addition to antidepressants for non-seasonal depression. PMID- 16043616 TI - Prophylactic mirtazapine may help to prevent post-stroke depression in people with good cognitive function. PMID- 16043617 TI - Cognitive behavioural therapy has short term but not long term benefits in people with residual symptoms of depression. PMID- 16043618 TI - Improving quality of primary care reduces depression and improves quality of life in adolescents. PMID- 16043619 TI - Review: cognitive behaviour therapy is beneficial in children with anxiety disorders. PMID- 16043620 TI - Adding mixed amphetamine salts to divalproex sodium improves ADHD symptoms in children with bipolar disorder and comorbid ADHD. PMID- 16043621 TI - Valproate reduces alcohol consumption in people with comorbid alcohol dependency and bipolar disorder. PMID- 16043622 TI - Lithium may reduce gambling severity in pathological gamblers with bipolar disorder. PMID- 16043623 TI - Adding a nurse-based intervention programme to usual care improves manic symptoms in people with bipolar disorder. PMID- 16043625 TI - Review: pharmacotherapy provides minimal improvements in the neuropsychiatric symptoms of dementia. PMID- 16043624 TI - Glycine and D-serine improve the negative symptoms of schizophrenia. PMID- 16043626 TI - Review: short term psychotherapy is an effective treatment for psychiatric disorders. PMID- 16043627 TI - Review: migrants are at increased risk of developing schizophrenia. PMID- 16043628 TI - Low intelligence increases risk of suicide. PMID- 16043629 TI - Cannabis use increases the risk of young people developing psychotic symptoms, particularly if already predisposed. PMID- 16043630 TI - Intoxicant use increased among Norwegian adolescents between 1992 and 2002, and attempted suicides increased among girls. PMID- 16043631 TI - A mathematical model of dynamic glioma-host interactions: receptor-mediated invasion and local proteolysis. AB - We present a mathematical model of glioma spread based on cellular movement by receptor-mediated haptotaxis, local proteolysis of healthy tissue components by glioma-derived proteinases, malignant proliferative enhancement and host up regulation of specific key extracellular matrix (ECM) components in response to the invading glioma. We subsequently consider the nature of glioma-host interactions as predicted by our model in order to test the hypothesis given in (Knott et al. (1998) that production of adhesive ECM components by the brain in response to the invading glioma may have the counter-intuitive effect of enhancing glioma invasion by assisting haptotactic migration. We suggest that host production of certain adhesive ECM chemicals can have a profound effect on both glioma invasion speed and the character of the glioma-host interface. In particular, we conclude that up-regulation of host ECM production in the vicinity of the glioma may produce a less diffuse glioma, providing clearer demarcation between glioma and healthy tissue, and thus improving the possibility of surgical resection within reasonable bounds. PMID- 16043632 TI - Topological constraints in nucleic acid hybridization kinetics. AB - A theoretical examination of kinetic mechanisms for forming knots and links in nucleic acid structures suggests that molecules involving base pairs between loops are likely to become topologically trapped in persistent frustrated states through the mechanism of 'helix-driven wrapping'. Augmentation of the state space to include both secondary structure and topology in describing the free energy landscape illustrates the potential for topological effects to influence the kinetics and function of nucleic acid strands. An experimental study of metastable complementary 'kissing hairpins' demonstrates that the topological constraint of zero linking number between the loops effectively prevents conversion to the minimum free energy helical state. Introduction of short catalyst strands that break the topological constraint causes rapid conversion to full duplex. PMID- 16043633 TI - Incorporation of non-nucleoside triphosphate analogues opposite to an abasic site by human DNA polymerases beta and lambda. AB - A novel class of non-nucleoside triphosphate analogues, bearing hydrophobic groups sterically similar to nucleosides linked to the alpha-phosphate but lacking the chemical functional groups of nucleic acids, were tested against six different DNA polymerases (polymerases). Human polymerases alpha, beta and lambda, and Saccharomyces cerevisiae polymerase IV, were inhibited with different potencies by these analogues. On the contrary, Escherichia coli polymerase I and HIV-1 reverse transcriptase were not. Polymerase beta incorporated these derivatives in a strictly Mn++-dependent manner. On the other hand, polymerase lambda could incorporate some alkyltriphosphate derivatives with both Mg++ and Mn++, but only opposite to an abasic site on the template strand. The active site mutant polymerase lambda Y505A showed an increased ability to incorporate the analogues. These results show for the first time that neither the base nor the sugar moieties of nucleotides are required for incorporation by family X DNA polymerases. PMID- 16043634 TI - Cloning and characterization of the Schizosaccharomyces pombe homologs of the human protein Translin and the Translin-associated protein TRAX. AB - Translin is a human octameric protein that specifically binds the single-stranded microsatellite repeats d(GT)n and the corresponding transcripts (GU)n. It also binds, with lesser affinities, other single-stranded G-rich DNA and RNA sequences. TRAX is a human protein that bears a homology to Translin and interacts with it. Translin and TRAX have been proposed to be involved in DNA recombination, chromosomal translocation and mRNA transport and translation. Both proteins are highly conserved in eukaryotes, including the fission yeast Schizosaccharomyces pombe, which is amenable to genetic analysis. Here, we report the first study of the S.pombe Translin and TRAX homologs. We have deleted the genes encoding Translin and TRAX in S.pombe and found that the proliferation of the mutant cells was slightly stimulated, suggesting that these genes are not essential for the fission yeast. We have also shown that the S.pombe Translin and TRAX interact. Biochemical analysis of the S.pombe Translin, which was cloned and expressed in Escherichia coli, revealed that it is octameric and that it selectively binds d(GT)n and d(GTT)n microsatellite repeats. However, unlike the human protein, it has much higher affinities for the homologous RNA sequences (GU)n and (GUU)n. These data suggest that the S.pombe Translin is primarily involved in functions related to RNA metabolism. PMID- 16043635 TI - MAO: a Multiple Alignment Ontology for nucleic acid and protein sequences. AB - The application of high-throughput techniques such as genomics, proteomics or transcriptomics means that vast amounts of heterogeneous data are now available in the public databases. Bioinformatics is responding to the challenge with new integrated management systems for data collection, validation and analysis. Multiple alignments of genomic and protein sequences provide an ideal environment for the integration of this mass of information. In the context of the sequence family, structural and functional data can be evaluated and propagated from known to unknown sequences. However, effective integration is being hindered by syntactic and semantic differences between the different data resources and the alignment techniques employed. One solution to this problem is the development of an ontology that systematically defines the terms used in a specific domain. Ontologies are used to share data from different resources, to automatically analyse information and to represent domain knowledge for non-experts. Here, we present MAO, a new ontology for multiple alignments of nucleic and protein sequences. MAO is designed to improve interoperation and data sharing between different alignment protocols for the construction of a high quality, reliable multiple alignment in order to facilitate knowledge extraction and the presentation of the most pertinent information to the biologist. PMID- 16043636 TI - Solution structure of the DNA-binding domain of RPA from Saccharomyces cerevisiae and its interaction with single-stranded DNA and SV40 T antigen. AB - Replication protein A (RPA) is a three-subunit complex with multiple roles in DNA metabolism. DNA-binding domain A in the large subunit of human RPA (hRPA70A) binds to single-stranded DNA (ssDNA) and is responsible for the species-specific RPA-T antigen (T-ag) interaction required for Simian virus 40 replication. Although Saccharomyces cerevisiae RPA70A (scRPA70A) shares high sequence homology with hRPA70A, the two are not functionally equivalent. To elucidate the similarities and differences between these two homologous proteins, we determined the solution structure of scRPA70A, which closely resembled the structure of hRPA70A. The structure of ssDNA-bound scRPA70A, as simulated by residual dipolar coupling-based homology modeling, suggested that the positioning of the ssDNA is the same for scRPA70A and hRPA70A, although the conformational changes that occur in the two proteins upon ssDNA binding are not identical. NMR titrations of hRPA70A with T-ag showed that the T-ag binding surface is separate from the ssDNA binding region and is more neutral than the corresponding part of scRPA70A. These differences might account for the species-specific nature of the hRPA70A-T-ag interaction. Our results provide insight into how these two homologous RPA proteins can exhibit functional differences, but still both retain their ability to bind ssDNA. PMID- 16043637 TI - Accelerated longitudinal decline of aerobic capacity in healthy older adults. AB - BACKGROUND: The ability of older persons to function independently is dependent largely on the maintenance of sufficient aerobic capacity and strength to perform daily activities. Although peak aerobic capacity is widely recognized to decline with age, its rate of decline has been estimated primarily from cross-sectional studies that may provide misleading, overly optimistic estimates of aging changes. METHODS AND RESULTS: To determine longitudinal rate of change in aerobic capacity and the influence of age, gender, and physical activity on these changes, we performed serial measurements of peak treadmill oxygen consumption (peak VO2) in 375 women and 435 men ages 21 to 87 years from the Baltimore Longitudinal Study of Aging, a community-dwelling cohort free of clinical heart disease, over a median follow-up period of 7.9 years. A linear mixed-effects regression model was used to calculate the predicted longitudinal 10-year rate of change in peak VO2, expressed in milliliters per minute, for each age decade from the 20s through the 70s after adjustment for self-reported leisure-time physical activity. A longitudinal decline in peak VO2 was observed in each of the 6 age decades in both sexes; however, the rate of decline accelerated from 3% to 6% per 10 years in the 20s and 30s to >20% per 10 years in the 70s and beyond. The rate of decline for each decade was larger in men than in women from the 40s onward. Similar longitudinal rates of decline prevailed when peak VO2 was indexed per kilogram of body weight or per kilogram of fat-free mass and in all quartiles of self-reported leisure-time physical activity. When the components of peak VO2 were examined, the rate of longitudinal decline of the oxygen pulse (ie, the O2 utilization per heart beat) mirrored that of peak VO2, whereas the longitudinal rate of heart rate decline averaged only 4% to 6% per 10 years, and accelerated only minimally with age. CONCLUSIONS: The longitudinal rate of decline in peak VO2 in healthy adults is not constant across the age span in healthy persons, as assumed by cross-sectional studies, but accelerates markedly with each successive age decade, especially in men, regardless of physical activity habits. The accelerated rate of decline of peak aerobic capacity has substantial implications with regard to functional independence and quality of life, not only in healthy older persons, but particularly when disease-related deficits are superimposed. PMID- 16043638 TI - Pilot study of rapid infusion of 2 L of 4 degrees C normal saline for induction of mild hypothermia in hospitalized, comatose survivors of out-of-hospital cardiac arrest. AB - BACKGROUND: Recent clinical studies have demonstrated that mild hypothermia (32 degrees C to 34 degrees C) induced by surface cooling improves neurological outcome after resuscitation from out-of-hospital cardiac arrest. Results from animal models suggest that the effectiveness of mild hypothermia could be improved if initiated as soon as possible after return of spontaneous circulation. Infusion of cold, intravenous fluid has been proposed as a safe, effective, and inexpensive technique to induce mild hypothermia after cardiac arrest. METHODS AND RESULTS: In 17 hospitalized survivors of out-of-hospital cardiac arrest, we determined the effect on temperature and hemodynamics of infusing 2 L of 4 degrees C cold, normal saline during 20 to 30 minutes into a peripheral vein with a high-pressure bag. Data on vital signs, electrolytes, arterial blood gases, and coagulation were collected before and after fluid infusion. Cardiac function was assessed by transthoracic echocardiography before fluid administration and 1 hour after infusion. Passive (fans, leaving patient uncovered) or active (cooling blankets, neuromuscular blockade) cooling measures were used to maintain mild hypothermia for 24 hours. Infusion of 2 L of 4 degrees C cold, normal saline resulted in a mean temperature drop of 1.4 degrees C 30 minutes after the initiation of infusion. Rapid infusion of fluid was not associated with clinically important changes in vital signs, electrolytes, arterial blood gases, or coagulation parameters. The initial mean ejection fraction was 34%, and fluid infusion did not affect ejection fraction or increase central venous pressure, pulmonary pressures, or left atrial filling pressures as assessed by echocardiography. Passive measures were ineffective in maintaining hypothermia compared with active measures. CONCLUSIONS: Infusion of 2 L of 4 degrees C cold, normal saline is safe and effective in rapidly lowering body temperature in survivors of out-of-hospital cardiac arrest. PMID- 16043639 TI - Real-time magnetic resonance imaging-guided stenting of aortic coarctation with commercially available catheter devices in Swine. AB - BACKGROUND: Real-time MR imaging (rtMRI) is now technically capable of guiding catheter-based cardiovascular interventions. Compared with x-ray, rtMRI offers superior tissue imaging in any orientation without ionizing radiation. Translation to clinical trials has awaited the availability of clinical-grade catheter devices that are both MRI visible and safe. We report a preclinical safety and feasibility study of rtMRI-guided stenting in a porcine model of aortic coarctation using only commercially available catheter devices. METHOD AND RESULTS: Coarctation stenting was performed wholly under rtMRI guidance in 13 swine. rtMRI permitted procedure planning, device tracking, and accurate stent deployment. "Active" guidewires, incorporating MRI antennas, improved device visualization compared with unmodified "passive" nitinol guidewires and shortened procedure time (26+/-11 versus 106+/-42 minutes; P=0.008). Follow-up catheterization and necropsy showed accurate stent deployment, durable gradient reduction, and appropriate neointimal formation. MRI immediately identified aortic rupture when oversized devices were tested. CONCLUSIONS: This experience demonstrates preclinical safety and feasibility of rtMRI-guided aortic coarctation stenting using commercially available catheter devices. Patients may benefit from rtMRI in the future because of combined device and tissue imaging, freedom from ionizing radiation, and the ability to identify serious complications promptly. PMID- 16043640 TI - Plasma oxidized low-density lipoprotein, a strong predictor for acute coronary heart disease events in apparently healthy, middle-aged men from the general population. AB - BACKGROUND: Oxidized LDL (oxLDL) is thought to play a key role in the inflammatory response in the arterial vessel wall. METHODS AND RESULTS: In a prospective, nested, case-control study, the association between plasma oxLDL and risk of an acute coronary heart disease (CHD) event was investigated in men without prevalent CHD or diabetes mellitus at baseline. Subjects came from 2 population-based MONICA/KORA Augsburg surveys conducted in the years 1989-1990 and 1994-1995 with follow-up in 1998 (mean+/-SD follow-up time, 5.6+/-2.6 years). OxLDL was determined by ELISA in 88 men with incident CHD and in 258 age- and survey-matched controls. Hazard ratios (HRs) were estimated from conditional logistic-regression models with matching for age and survey. Baseline mean plasma oxLDL concentrations were significantly higher in subjects who subsequently experienced an event compared with controls (mean+/-SD, 110+/-32 versus 93+/-28 U/L; P< or =0.001). After adjustment for smoking, hypertension, obesity, physical activity, education, and alcohol consumption, the HR for a future CHD event in a comparison of the upper tertile of the oxLDL distribution with the lower tertile was 4.25 (95% confidence interval, 2.09 to 8.63; P<0.001). Plasma oxLDL was the strongest predictor of CHD events compared with a conventional lipoprotein profile and other traditional risk factors for CHD. When both oxLDL and C reactive protein were simultaneously assessed in the same model, they still predicted future CHD events even after multivariable adjustment. CONCLUSIONS: Elevated concentrations of oxLDL are predictive of future CHD events in apparently healthy men. Thus, oxLDL may represent a promising risk marker for clinical CHD complications and should be evaluated in further studies. PMID- 16043641 TI - Cerebral vascular dysfunction in methionine synthase-deficient mice. AB - BACKGROUND: Methionine synthase (MS) catalyzes the folate-dependent remethylation of homocysteine to methionine. We tested the hypothesis that deficiency of MS impairs endothelial function in mice heterozygous for disruption of the Mtr gene, which encodes MS. METHODS AND RESULTS: Plasma total homocysteine was similar in wild-type (Mtr(+/+)) and heterozygous (Mtr(+/-)) mice fed a control diet (4.5+/ 0.3 and 5.3+/-0.4 micromol/L, respectively) and mildly elevated in Mtr(+/+) and Mtr(+/-) mice fed a low-folate (LF) diet (7.5+/-0.7 and 9.6+/-1.2 micromol/L, respectively; P<0.001 versus control diet). Dilatation of cerebral arterioles to the endothelium-dependent dilator, acetylcholine (10 micromol/L) was blunted in Mtr(+/-) mice compared with Mtr(+/+) mice fed the control diet (21+/-4 versus 32+/-4%; P<0.05). Both Mtr(+/+) and Mtr(+/-) mice exhibited impaired dilatation of cerebral arterioles to acetylcholine when they were fed the LF diet (12+/-2 and 14+/-2%, respectively; P<0.01 versus Mtr(+/+) mice fed the control diet). Elevated levels of superoxide and hydrogen peroxide were detected by confocal microscopy in cerebral arterioles of Mtr(+/-) mice fed the control diet and in both Mtr(+/+) and Mtr(+/-) mice fed the LF diet. CONCLUSIONS: These findings demonstrate that defective homocysteine remethylation caused by deficiency of either MS or folate produces oxidative stress and endothelial dysfunction in the cerebral microcirculation of mice. PMID- 16043642 TI - Kruppel-like factor 2 as a novel mediator of statin effects in endothelial cells. AB - BACKGROUND: Although 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) are known to modulate endothelial function, the transcriptional mechanisms underlying these effects are incompletely understood. We hypothesized that Lung-Kruppel-like factor (LKLF/KLF2), a novel and potent regulator of endothelial gene expression, may mediate the downstream effects of statins. Here we report that statin-induced expression of endothelial NO synthase (eNOS) and thrombomodulin is KLF2 dependent. METHODS AND RESULTS: KLF2 mRNA was induced by treatment with multiple statins in a concentration-dependent manner. Multiple lines of evidence suggest that this induction is dependent on inhibition of the Rho pathway and requires de novo transcription. Furthermore, promoter deletion and mutational analyses suggest that mevastatin induced KLF2 promoter activity through a single myocyte enhancer factor binding site. Finally, small-interfering RNA-mediated knockdown of KLF2 strongly attenuated the ability of mevastatin to increase eNOS and thrombomodulin accumulation in endothelial cells. CONCLUSIONS: Taken together, these observations indicate that statin-dependent induction of eNOS and thrombomodulin requires KLF2 and thereby provides a novel molecular target for modulating endothelial function in vascular disease. PMID- 16043643 TI - Overexpression of mitochondrial transcription factor a ameliorates mitochondrial deficiencies and cardiac failure after myocardial infarction. AB - BACKGROUND: Mitochondrial DNA (mtDNA) copy number is decreased not only in mtDNA mutation diseases but also in a wide variety of acquired degenerative and ischemic diseases. Mitochondrial transcription factor A (TFAM) is essential for mtDNA transcription and replication. Myocardial mtDNA copy number and TFAM expression both decreased in cardiac failure. However, the functional significance of TFAM has not been established in this disease state. METHODS AND RESULTS: We have now addressed this question by creating transgenic (Tg) mice that overexpress human TFAM gene and examined whether TFAM could protect the heart from mtDNA deficiencies and attenuate left ventricular (LV) remodeling and failure after myocardial infarction (MI) created by ligating the left coronary artery. TFAM overexpression could ameliorate the decrease in mtDNA copy number and mitochondrial complex enzyme activities in post-MI hearts. Survival rate during 4 weeks of MI was significantly higher in Tg-MI than in wild-type (WT) littermates (WT-MI), although infarct size was comparable. LV cavity dilatation and dysfunction were significantly attenuated in Tg-MI. LV end-diastolic pressure was increased in WT-MI, and it was also reduced in Tg-MI. Improvement of LV function in Tg-MI was accompanied by a decrease in myocyte hypertrophy, apoptosis, and interstitial fibrosis as well as oxidative stress in the noninfarcted LV. CONCLUSIONS: Overexpression of TFAM inhibited LV remodeling after MI. TFAM may provide a novel therapeutic strategy of cardiac failure. PMID- 16043644 TI - Genetic analysis of the interleukin-18 system highlights the role of the interleukin-18 gene in cardiovascular disease. AB - BACKGROUND: Interleukin (IL)-18 plays a key role in atherosclerosis and its complications. The present study investigated the genetic variability of 4 genes of the IL-18 system-IL18, IL18R1, IL18RAP, and IL18BP-in relation to circulating IL-18 levels and cardiovascular mortality. METHODS AND RESULTS: Twenty-two polymorphisms were genotyped in 1288 patients with coronary artery disease prospectively followed up during a median period of 5.9 years. The end point was death from cardiovascular causes (n=142). Baseline IL-18 levels were predictive of cardiovascular deaths occurring during < or =4 years of follow-up (HR=2.96, 95% CI 1.54 to 5.70, P=0.001 for the top compared with the bottom quartile) but not of later deaths. Haplotypes of the IL18 gene were associated with IL-18 levels (P=0.002) and cardiovascular mortality (P=0.006) after adjustment for cardiovascular risk factors. The same haplotype was associated with both a 9% lowering effect on IL-18 levels and a protective effect on risk (HR=0.57, 95% CI 0.36 to 0.92). IL18 haplotypes explained only 2% of IL-18 variability. Adjustment for baseline IL-18 levels abolished the association of haplotypes with cardiovascular risk. The haplotype associated with phenotypes was the only one carrying the minor allele of the IL18/A+183G polymorphism located in the 3'untranslated region and potentially affecting mRNA stability. The other genes of the system were not related to IL-18 levels or cardiovascular outcome. CONCLUSIONS: Variations of the IL18 gene consistently influence circulating levels of IL-18 and clinical outcome in patients with coronary artery disease, which supports the hypothesis of a causal role of IL-18 in atherosclerosis and its complications. PMID- 16043645 TI - Prevalence of cardiovascular disease risk factor clustering among the adult population of China: results from the International Collaborative Study of Cardiovascular Disease in Asia (InterAsia). AB - BACKGROUND: The prevalence of cardiovascular disease (CVD) risk factors has been increasing in China. METHODS AND RESULTS: We examined the prevalence of CVD risk factor clustering among Chinese adults aged 35 to 74 years with data from the International Collaborative Study of Cardiovascular Disease in Asia (InterAsia), a cross-sectional survey of a nationally representative sample (n=14 690) conducted during 2000 to 2001 and compared these data with those of US adults from the National Health and Nutrition Examination Survey of 1999 to 2000. Overall, 80.5%, 45.9%, and 17.2% of Chinese adults had > or =1, > or =2, and > or =3 modifiable CVD risk factors (dyslipidemia, hypertension, diabetes, cigarette smoking, and overweight), respectively. By comparison, 93.1%, 73.0%, and 35.9% of US adults had > or =1, > or =2, and > or =3 of these risk factors, respectively. In a multivariate model including age, sex, and area of residence, the odds ratio (95% confidence interval [CI]) of having > or =1, > or =2, and > or =3 CVD risk factors versus none of the studied risk factors was 2.61 (95% CI, 2.09 to 3.27), 3.55 (95% CI, 2.77 to 4.54), and 4.97 (95% CI, 3.67 to 6.74), respectively, for Chinese adults 65 to 74 years old versus 35 to 44 years old; 3.65 (95% CI, 3.21 to 4.15), 4.67 (95% CI, 4.06 to 5.38), and 5.60 (95% CI, 4.70 to 6.67), respectively, for men compared with women; 1.18 (95% CI, 1.07 to 1.30), 1.34 (95% CI, 1.21 to 1.50), and 1.84 (95% CI, 1.60 to 2.12), respectively, for urban compared with rural residents; and 1.98 (95% CI, 1.76 to 2.22), 2.75 (95% CI, 2.42 to 3.13), and 4.36 (95% CI, 3.68 to 5.18), respectively, for residents of northern compared with southern China. CONCLUSIONS: Clustering of CVD risk factors is common in China. Prevention, detection, and treatment of CVD risk factor clustering should be an important component of a national strategy to reduce the increasing burden of CVD in China. PMID- 16043646 TI - Mechanisms of cardiac depression caused by lipoteichoic acids from Staphylococcus aureus in isolated rat hearts. AB - BACKGROUND: Lipoteichoic acid (LTA) represents a major virulence factor in gram positive sepsis. METHODS AND RESULTS: In the present study we perfused isolated rat hearts for 180 minutes with highly purified LTA from Staphylococcus aureus. A progressive decline of left ventricular contractile function paralleled by the expression of myocardial tumor necrosis factor-alpha (TNF-alpha) mRNA and protein as well as the release of TNF-alpha into the perfusate was observed in LTA perfused hearts. Employment of an anti-TNF-alpha antibody completely prevented the loss in contractile function. When CD14, a prominent pathogen recognition receptor, was blocked by a specific antibody, induction of TNF-alpha mRNA and protein release as well as the associated cardiodepression was diminished in response to LTA. Synthesis of TNF-alpha protein was located to interstitial cells of LTA-challenged hearts as detected by immunohistochemistry. Besides progressive cardiodepression, coronary perfusion pressure (CPP) was moderately increased in LTA-perfused hearts. This was accompanied by the release of thromboxane A2 (TXA2) into the perfusate and the induction of cyclooxygenase (Cox)-2 mRNA and protein in the myocardium. Blocking of TXA2 by the nonspecific Cox inhibitor indomethacin, the thromboxane receptor antagonist daltroban, or the selective Cox 2 inhibitor NS-398 prevented the increase in CPP. CONCLUSIONS: LTA causes cardiac depression by activating myocardial TNF-alpha synthesis via CD14 and induces coronary vascular disturbances by activating Cox-2-dependent TXA2 synthesis. These phenomena may contribute to cardiac depression in gram-positive sepsis. PMID- 16043647 TI - Upregulation of nitric oxide production in vascular endothelial cells by all trans retinoic acid through the phosphoinositide 3-kinase/Akt pathway. AB - BACKGROUND: A natural retinoid all-trans retinoic acid (ATRA) contains various beneficial effects on vasculature, including suppression of neointima formation after balloon injury. However, little is known about the effects of ATRA on vascular endothelial function. We therefore studied its role in nitric oxide (NO) production of vascular endothelial cells (ECs). METHODS AND RESULTS: Human dermal microvascular ECs, human umbilical vein ECs, and SV40-transformed rat lung vascular ECs were incubated with or without ATRA (1 micromol/L) for 48 hours. Their NO production was determined with the use of a fluorescent NO indicator, diaminofluorescein-2 diacetate. ATRA significantly increased their basal as well as acetylcholine-induced NO production. Treatment with Nomega-nitro-L-arginine methyl ester or carboxy-PTIO suppressed their fluorescence. Increase of NO production was also observed by incubation with retinoic acid receptor (RAR) agonist Am580. ATRA-induced NO increase was abolished by coincubation with RAR antagonist LE540. Moreover, the NO increase was completely inhibited by the phosphoinositide 3-kinase (PI3K) inhibitor wortmannin and LY294002. ATRA as well as Am580 enhanced endothelial NO synthase (eNOS) phosphorylation at Ser-1177 as well as Akt phosphorylation at Ser-473 without changing their protein expression. Overexpression of dominant-negative Akt inhibited the eNOS phosphorylation. Moreover, ATRA increased PI3K activity as well as PI3K catalytic subunit p110beta protein expression, which was completely inhibited by LE540 treatment. Real-time polymerase chain reaction analyses demonstrated that ATRA increased PI3K catalytic subunit p110beta mRNA expression without affecting its stability. Finally, ATRA-induced NO increase was observed in COS-1 cells transfected with wild-type eNOS and RARalpha, but not with mutated eNOS whose Ser-1177 was substituted. CONCLUSIONS: ATRA increases NO production by eNOS phosphorylation through RAR-mediated PI3K/Akt pathway activation in vascular ECs and possibly plays beneficial roles in vascular endothelium. Retinoids may therefore be candidates as novel therapeutic agents against vascular disorders with endothelial damage. PMID- 16043648 TI - Current management of severe congenital mitral stenosis: outcomes of transcatheter and surgical therapy in 108 infants and children. AB - BACKGROUND: Severe congenital mitral stenosis (MS) is a rare anomaly that is frequently associated with additional left heart obstructions. Anatomic treatments for congenital MS include balloon mitral valvuloplasty (BMVP), surgical mitral valvuloplasty (SMVP), and mitral valve replacement (MVR), although the optimal therapeutic strategy is unclear. METHODS AND RESULTS: Between 1985 and 2003, 108 patients with severe congenital MS underwent BMVP or surgical intervention at a median age of 18 months (range 1 month to 17.9 years). Anatomic subtypes of MS were "typical" congenital MS in 78 patients, supravalvar mitral ring in 46, parachute mitral valve in 28, and double-orifice mitral valve in 11, with multiple types in approximately 50% of patients. Additional left heart anomalies were present in 82 patients (76%). The first MS intervention was BMVP in 64 patients, SMVP in 33, and MVR in 11. BMVP decreased peak and mean MS gradients by a median of 33% and 38%, respectively (P<0.001), but was complicated by significant mitral regurgitation in 28%. Cross-sectional follow-up was obtained at 4.8+/-4.2 years. Overall, Kaplan-Meier survival was 92% at 1 month, 84% at 1 year, and 77% at 5 years, with 69% 5-year survival during the first decade of our experience and 87% since (P=0.09). Initial MVR and younger age were associated with worse survival. Survival free from failure of biventricular repair or mitral valve reintervention was 55% at 1 year among patients who underwent BMVP and 69% among patients who underwent supravalvar mitral ring resection initially. Among patients who underwent BMVP, survival free from failure of biventricular repair or MVR was 79% at 1 month and 55% at 5 years, with worse outcome in younger patients and those who developed significant postdilation mitral regurgitation. CONCLUSIONS: BMVP effectively relieves left ventricular inflow obstruction in most infants and children with severe congenital MS who require intervention. However, surgical resection is preferable in patients with MS due to a supravalvar mitral ring. Five-year survival is relatively poor in patients with severe congenital MS, with worse outcomes in infants and patients undergoing MVR, but has improved in our more recent experience. Many patients have undergone second procedures for either recurrent/residual MS or mitral regurgitation resulting from dilation-related disruption of the mitral valve apparatus. PMID- 16043649 TI - Coagulation assays. PMID- 16043650 TI - Images in cardiovascular medicine. Detection of cardiac metastasis by positron emission tomography-computed tomography. PMID- 16043651 TI - Images in cardiovascular medicine. Early tissue distribution of bone marrow mononuclear cells after transcoronary transplantation in a patient with acute myocardial infarction. PMID- 16043652 TI - Images in cardiovascular medicine. Giant aneurysm of the right coronary artery compressing the right heart. PMID- 16043653 TI - Letter regarding article by Nasir et al, "Electrocardiographic features of arrhythmogenic right ventricular dysplasia/cardiomyopathy according to disease severity: a need to broaden diagnostic criteria". PMID- 16043654 TI - Letter regarding article by Thambo et al, "Detrimental ventricular remodeling in patients with congenital complete heart block and chronic right ventricular apical pacing". PMID- 16043655 TI - Surgical versus alcohol septal ablation for hypertrophic obstructive cardiomyopathy: the pendulum swings. PMID- 16043656 TI - Our passive lifestyle, our toxic diet, and the atherogenic/diabetogenic metabolic syndrome: can we afford to be sedentary and unfit? PMID- 16043657 TI - Effective interventions to reduce smoking-induced heart disease around the world: time to act. PMID- 16043658 TI - Inhibition of atherogenesis in BLT1-deficient mice reveals a role for LTB4 and BLT1 in smooth muscle cell recruitment. AB - BACKGROUND: It is known that 5-lipoxygenase and its product, leukotriene B4 (LTB4), are highly expressed in several human pathologies, including atherosclerotic plaque. LTB(4) signals primarily through its high-affinity G protein-coupled receptor BLT1, which is expressed on specific leukocyte subsets. BLT1 receptor expression and function on other atheroma-associated cell types is unknown. METHODS AND RESULTS: To directly assess the role of the LTB4-BLT1 pathway in atherogenesis, we bred BLT1(-/-) mice into the atherosclerosis susceptible apoE(-/-) strain. Compound-deficient apoE(-/-)/Blt1(-/-) mice fed a Western-type diet had a marked reduction in plaque formation compared with apoE( /-) controls. Immunohistochemical analysis of atherosclerotic lesions in compound deficient mice revealed a striking decrease in smooth muscle cells (SMCs) and significant decreases in macrophages and T cells. We report here novel evidence of the expression and function of BLT1 on vascular SMCs. LTB4 triggered SMC chemotaxis, which was pertussis toxin sensitive in Blt1(+/+) SMCs and absent in Blt1(-/-) cells, suggesting that BLT1 was the dominant receptor mediating effector functions through a G protein-coupled signaling pathway. Furthermore, BLT1 colocalized with SMCs in human atherosclerotic lesions. CONCLUSIONS: These new findings extend the role of inducible BLT1 to nonleukocyte populations and suggest an important target for intervention to modulate the response to vascular injury. PMID- 16043659 TI - Recent advances in cardiac development with therapeutic implications for adult cardiovascular disease. PMID- 16043660 TI - Contribution of 20-HETE to augmented myogenic constriction in coronary arteries of endothelial NO synthase knockout mice. AB - Previous studies suggested an important role for 20-HETE in the regulation of myogenic responses. Thus, pressure-diameter relationships were investigated in isolated, cannulated coronary arteries (approximately 100 microm) from male endothelial NO synthase knockout (eNOS-KO) and wild-type (WT) mice. All arteries constricted in response to step increases in perfusate pressure from 20 to 100 mm Hg. This constriction was significantly enhanced from 40 to 100 mm Hg in arteries of eNOS-KO compared with those of WT mice. For example, at 60 and 100 mm Hg, respectively, the normalized diameter (expressed as a percentage of the corresponding passive diameter) of arteries of eNOS-KO mice was 10% and 12% smaller than that of WT mice. Removal of the endothelium did not significantly affect the responses of vessels from either strain of mice. However, N methylsulfonyl-12,12-dibromododec-11-enamide (5x10(-6) M), an inhibitor of cytochrome P-450 (CYP)/omega-hydroxylase, significantly attenuated the greater myogenic constriction of arteries from eNOS-KO mice by approximately 12% at each pressure step but did not significantly affect responses of those from WT mice, leading to a comparable myogenic response in the 2 strains. Western blot analysis demonstrated a comparable CYP4A protein content in coronary arteries of the 2 strains of mice. However, production of 20-HETE, measured by fluorescent high performance liquid chromatography assay was approximately 2.7-fold greater in eNOS-KO compared to WT mice. Thus, as a function of eNOS deficiency, the enhanced coronary artery constriction to pressure is attributable to an increased activity of omega-hydroxylase, which, consequently, increases the synthesis of 20-HETE in vascular smooth muscle. PMID- 16043661 TI - Combined angiotensin II type 1 and type 2 receptor blockade on vascular remodeling and matrix metalloproteinases in resistance arteries. AB - We investigated the role of angiotensin II type 1 (AT1) and AT2 receptors, matrix metalloproteinases (MMPs), and extracellular matrix (ECM) components involved in vascular remodeling of resistance arteries induced by angiotensin II (Ang II). Sprague-Dawley rats received Ang II (120 ng/kg per minute SC) +/- the AT1 antagonist losartan (10 mg/kg per day PO), the AT1/AT2 antagonist Sar1-Ile8-Ang II (Sar-Ile; 10 microg/kg per minute SC), or hydralazine (25 mg/kg per day PO) for 7 days. Structure and mechanical properties of small mesenteric arteries were evaluated on a pressurized myograph. Ang II increased growth index (+21%), which was partially decreased by losartan (-11%) and abrogated by Sar-Ile. Hydralazine markedly increased growth index (+32%) despite systolic blood pressure (BP) lowering, suggesting a BP-independent effect of Ang II on vascular growth. Elastic modulus was increased by Sar-Ile compared with Ang II and control. Vascular type I collagen was reduced (P<0.05), whereas fibronectin increased significantly with Sar-Ile. Vascular tissue inhibitor of metalloproteinase-2 binding to MMP-2 was abrogated by Sar-Ile, but MMP-2 activity was significantly increased compared with losartan, Ang II, and controls. Thus, AT1 blockade exerted antigrowth effects and reduced stiffness of small resistance arteries by decreasing nonelastic fibrillar components (collagen and fibronectin). Concomitant AT1/AT2 blockade prevented growth, reduced collagen type I and elastin deposition but increased vascular stiffness, fibronectin, and MMP-2 activity. These results demonstrate opposing roles of AT1 receptors that increase fibronectin and vascular stiffness and AT2 receptors that decrease MMP-2 and increase elastin. Changes in vascular wall mechanics, ECM deposition, and MMP activity are thus modulated differentially by Ang II receptors. PMID- 16043662 TI - Aldosterone stimulates matrix metalloproteinases and reactive oxygen species in adult rat ventricular cardiomyocytes. AB - Matrix metalloproteinases (MMPs), aldosterone, and reactive oxygen species (ROS) are implicated in myocardial remodeling. Although ROS, cytokines, and neurohormones regulate MMP in cardiac fibroblasts, it is unknown whether aldosterone regulates MMP in cardiomyocytes. Therefore, we tested the hypothesis that aldosterone regulates MMP in cultured adult rat ventricular myocytes (ARVMs). ARVMs were treated with aldosterone for 24 hours, and MMP-2 and MMP-9 activities were measured by zymography. Aldosterone (50 nmol/L) increased MMP-2 (43+/-5%) and MMP-9 (55+/-15%; P<0.001 for both) activities. Pretreatment with spironolactone (100 nmol/L) abolished the aldosterone-induced increase in MMP activities. Aldosterone (50 nmol/L; 30 minutes) increased mitogen/extracellular signal-regulated kinase (MEK) (31+/-3%) and extracellular signal-regulated kinase 1/2 (ERK1/2; 41+/-7%; P<0.001 for both) phosphorylation. U0126 (10 micromol/L), an MEK1/2 inhibitor, abolished the aldosterone-induced increase in MMP activities. Aldosterone increased intracellular ROS as assessed by dichlorofluorescein diacetate (27+/-4%; P<0.05). This increase was inhibited by apocynin, an NADPH oxidase inhibitor. Apocynin likewise inhibited aldosterone induced ERK1/2 phosphorylation and the increase in MMP activities. Furthermore, the antioxidants MnTMPyP and N-acetylcysteine inhibited the aldosterone-induced increase in ERK1/2 phosphorylation and MMP activities, respectively. Protein kinase C (PKC) is implicated in the nongenomic effects of aldosterone. To test the role of PKC, ARVMs were pretreated with chelerythrine, a PKC inhibitor. Chelerythrine prevented the aldosterone-induced increase in ERK1/2 phosphorylation and MMP activities. Thus, aldosterone induces MMP activity in ARVM via activation of the mineralocorticoid receptor, PKC, and ROS-dependent activation of the MEK/ERK pathway. NADPH oxidase is a likely source of ROS in this system. PMID- 16043663 TI - Local renal aldosterone system and its regulation by salt, diabetes, and angiotensin II type 1 receptor. AB - CYP11B2 is the enzyme responsible for aldosterone synthesis mainly in the adrenal gland. In this study, we hypothesized that CYP11B2 gene, protein, and aldosterone are produced locally in kidney and regulated by low salt intake, angiotensin II type 1 (AT1) receptor and insulin-deficient diabetes hyperglycemia. We used real time RT-PCR, immunohistochemistry staining, and microdialysis techniques to monitor changes in renal CYP11B2 mRNA and protein and aldosterone production in normal, adrenalectomized, or streptozotocin-induced insulin-deficient diabetic hyperglycemic rats. In normal kidney, CYP11B2 mRNA and protein were localized mainly in the renal cortex and upregulated by angiotensin II and low salt intake. The angiotensin II effect was reversed by AT1 receptor blocker valsartan. Immunohistochemistry staining demonstrated presence of CYP11B2 in glomeruli. Although aldosterone was absent in plasma of adrenalectomized rats, it was present in renal interstitium and tissue. Diabetes increased renal cortical and total kidney CYP11B2 mRNA and protein. Lowering blood glucose with insulin decreased total renal CYP11B2 mRNA and protein. Despite lack of significant changes in blood glucose, valsartan treatment caused significant reduction in renal CYP11B2 mRNA and protein. In presence of diabetes, there was an increase in CYP11B2 immunostaining in glomeruli and proximal tubules. This expression was abrogated with insulin or valsartan treatment. These results demonstrate the presence of all components of local renal aldosterone system. This system is physiologically active because it is regulated by angiotensin II and low salt intake. In insulin-deficient diabetes hyperglycemia rat model, glucose, insulin, and AT1 receptor modulate CYP11B2 expression in the kidney. PMID- 16043664 TI - Cardiovascular risk in relation to alpha-adducin Gly460Trp polymorphism and systolic pressure: a prospective population study. AB - Preliminary evidence from 1 case-control study suggested that in hypertensive patients, the alpha-adducin 460Trp allele might be associated with a 2-fold higher risk of coronary heart disease. In a prospective population study, we investigated whether the alpha-adducin Gly460Trp polymorphism predicted mortality and morbidity. From August 1985 until July 2003, we randomly recruited 2235 Belgian residents. We obtained information on vital status (until July 1, 2004) and the incidence of events via registries and repeat examinations (median 3). In Cox regression, before and after adjustment for other risk factors, we found strong interaction between systolic blood pressure at baseline, analyzed as a continuous variable, and the alpha-adducin polymorphism in relation to total (P=0.01) and cardiovascular mortality (P=0.007) and all cardiovascular (P=0.003), cardiac (P=0.001), and coronary events (P=0.03). The hazard ratio for total mortality associated with the Trp allele relative to GlyGly homozygosity was 2.30 (95% confidence interval, 1.12 to 4.72; P=0.02) in patients with stage-2 systolic hypertension (> or =160 mm Hg) and 0.88 (0.61 to 1.26; P=0.48) in the other participants. For all cardiovascular complications, these estimates were 2.94 (1.28 to 6.74; P=0.01) and 0.83 (0.58 to 1.20; P=0.32), respectively. For all cardiovascular events, the positive predictive value and the attributable risk associated with the Trp allele in patients with stage-2 systolic hypertension were 76.9% and 44.3%, respectively. In conclusion, the alpha-adducin Gly460Trp polymorphism, in combination with systolic blood pressure, is a strong predictor of cardiovascular mortality and morbidity. PMID- 16043665 TI - The ABCDs of treating the most prevalent childhood disease. AB - Dental caries is a potentially preventable infectious disease that, untreated, can cause significant morbidity requiring costly treatment. The Access to Baby and Child Dentistry (ABCD) program increases access to prevention and early treatment for Medicaid-eligible children aged younger than 6 years in Washington State. The program is a partnership of Washington Dental Service Foundation, the University of Washington School of Pediatric Dentistry, the state Medical Assistance Administration, and local stakeholders. Through training in pediatric dental techniques and enhanced reimbursement, ABCD equips and encourages dentists to serve young Medicaid patients. Family counseling increases oral care compliance and reduces no-show rates. Program evaluation demonstrates changing attitudes and behavior among participating families and dentists, with more young, low-income Washington children receiving oral health care. PMID- 16043666 TI - Barriers to care-seeking for children's oral health among low-income caregivers. AB - OBJECTIVES: We identified psychosocial, structural, and cultural barriers to seeking dental care among nonutilizing caregivers of Medicaid-enrolled children. METHODS: We used Medicaid utilization records to identify utilizing and nonutilizing African American and White caregivers of Medicaid-enrolled children in Jefferson County, Kentucky. We conducted 8 focus groups (N=76) with a stratified random sample of responding caregivers; transcripts were qualitatively analyzed. RESULTS: Psychosocial factors associated with utilization included oral health beliefs, norms of caregiver responsibility, and positive caregiver dental experiences. Utilizing groups reported higher education; health beliefs included identifying oral health with overall health and professional preventive dental care with caregiver responsibility for children's overall health. These beliefs may mediate shared structural barriers, including transportation, school absence policies, discriminatory treatment, and difficulty locating providers who accept Medicaid. Expectation of poor oral health among some low-income caregivers was among factors identified with nonutilization. CONCLUSIONS: Disadvantaged caregivers reported multiple barriers to accessing dental care for their children. Providers, Medicaid administrators, and schools must coordinate steps to encourage caregiver-controlled dental care, build trust, and link professional preventive dental care with caregiver responsibility for children's overall health. PMID- 16043667 TI - Preventive dental care and unmet dental needs among low-income children. AB - OBJECTIVES: We examined the ways in which levels of preventive dental care and unmet dental needs varied among subgroups of low-income children. METHODS: Data were drawn from the 2002 National Survey of America's Families. We conducted bivariate and multivariate analyses, including logistic regression analyses, to assess relationships between socioeconomic, demographic, and health factors and receipt of preventive dental care and unmet dental needs. RESULTS: More than half of low-income children without health insurance had no preventive dental care visits. Levels of unmet dental needs among low-income children who had private health insurance coverage but no dental benefits were similar to those among uninsured children. Children of parents whose mental health was rated as poor were twice as likely to have unmet dental needs as other children. CONCLUSIONS: Additional progress toward improving the dental health of low-income children depends on identifying and responding to factors limiting both the demand for and the supply of dental services. In particular, it appears that expanding access to dental benefits is key to improving the oral health of this population. PMID- 16043668 TI - Timeliness of childhood immunizations: a state-specific analysis. AB - OBJECTIVE: We examined the timeliness of vaccine administration among children aged 24 to 35 months for each state of the United States and the District of Columbia. METHODS: We analyzed the timeliness of vaccinations in the 2000-2002 National Immunization Survey. We used a modified Bonferroni adjustment to compare a reference state with all other states. RESULTS: Receipt of all vaccinations as recommended ranged from 2% (Mississippi) to 26% (Massachusetts), with western states having less timeliness than eastern states. CONCLUSIONS: Vaccination coverage measures usually focus on the number of vaccinations accumulated by specified ages. Our analysis of timeliness of administration shows that children rarely receive all vaccinations as recommended. State health departments can use timeliness of vaccinations along with other measures to determine children's susceptibility to vaccine-preventable diseases and to evaluate the quality of vaccination programs. States can use the modified Bonferroni comparison to appropriately compare their results with other states. PMID- 16043669 TI - Mortality in US Army Gulf War veterans exposed to 1991 Khamisiyah chemical munitions destruction. AB - OBJECTIVES: We investigated whether US Army Gulf War veterans who were potentially exposed to nerve agents during the March 1991 weapons demolitions at Khamisiyah, Iraq, are at increased risk of cause-specific mortality. METHODS: The cause-specific mortality of 100487 exposed US Army Gulf War veterans was compared with that of 224980 unexposed US Army Gulf War veterans. Exposure was determined with the Department of Defense 2000 plume model. Relative risk estimates were derived from Cox proportional hazards models. RESULTS: The risks of most disease related mortality were similar for exposed and unexposed veterans. However, exposed veterans had an increased risk of brain cancer deaths (relative risk [RR]=1.94; 95% confidence interval [CI]=1.12, 3.34). The risk of brain cancer death was larger among those exposed 2 or more days than those exposed 1 day when both were compared separately to all unexposed veterans (RR=3.26; 95% CI=1.33, 7.96; RR=1.72; 95% CI=0.95,3.10, respectively). CONCLUSIONS: Exposure to chemical munitions at Khamisiyah may be associated with an increased risk of brain cancer death. Additional research is required to confirm this finding. PMID- 16043670 TI - Limited English proficiency and breast and cervical cancer screening in a multiethnic population. AB - OBJECTIVES: We examined the relationship between ability to speak English and receipt of Papanicolaou tests, clinical breast examinations, and mammography in a multiethnic group of women in the United States. METHODS: We used longitudinal data from the Study of Women Across the Nation to examine receipt of breast and cervical cancer screening among Chinese, Japanese, Hispanic, and White women who reported reading and speaking (1) only a language other than English, (2) another language more fluently than English, or (3) only English or another language and English with equal fluency. Logistic regression was used to analyze the data. RESULTS: Reading and speaking only a language other than English and reading and speaking another language more fluently than English, were significantly and negatively associated with receipt of breast and cervical cancer screening in unadjusted models. Although these findings were attenuated in adjusted models, not speaking English well or at all remained negatively associated with receipt of cancer screening. CONCLUSIONS: These findings suggest that language barriers contribute to health disparities by impeding adequate health communication. PMID- 16043671 TI - Health care expenditures of immigrants in the United States: a nationally representative analysis. AB - OBJECTIVES: We compared the health care expenditures of immigrants residing in the United States with health care expenditures of US-born persons. METHODS: We used the 1998 Medical Expenditure Panel Survey linked to the 1996-1997 National Health Interview Survey to analyze data on 18398 US-born persons and 2843 immigrants. Using a 2-part regression model, we estimated total health care expenditures, as well as expenditures for emergency department (ED) visits, office-based visits, hospital-based outpatient visits, inpatient visits, and prescription drugs. RESULTS: Immigrants accounted for $39.5 billion (SE=$4 billion) in health care expenditures. After multivariate adjustment, per capita total health care expenditures of immigrants were 55% lower than those of US-born persons ($1139 vs $2546). Similarly, expenditures for uninsured and publicly insured immigrants were approximately half those of their US-born counterparts. Immigrant children had 74% lower per capita health care expenditures than US-born children. However, ED expenditures were more than 3 times higher for immigrant children than for US-born children. CONCLUSIONS: Health care expenditures are substantially lower for immigrants than for US-born persons. Our study refutes the assumption that immigrants represent a disproportionate financial burden on the US health care system. PMID- 16043672 TI - An analysis of messages about tobacco in military installation newspapers. AB - OBJECTIVES: We sought to gauge the relative attention that tobacco control receives in military newspapers by comparing coverage of tobacco use with that of other health topics of importance to the military. METHODS: We examined tobacco related articles and industry advertisements in 793 newspapers published during 1 year at 16 representative military installations (4 Air Force, 6 Army, 2 Marine, 4 Navy). Newspaper content was coded with a standardized coding manual developed through previous research. RESULTS: Tobacco use received the fewest instances of coverage and the least print space in military installation newspapers of all the health topics examined. The primary "message frame" used in tobacco control articles was that smokers are putting themselves at health risk, a theme that has not been found to have a strong effect on smokers. Nearly 10% of the newspapers contained tobacco advertisements. CONCLUSIONS: Tobacco control messages are underrepresented in military installation newspapers compared with other health issues. Furthermore, military newspapers send mixed messages to military personnel by providing advertisements for tobacco while also claiming that tobacco use is harmful. PMID- 16043673 TI - Goldilocks and menopause. PMID- 16043674 TI - American pain society recommendations for improving the quality of acute and cancer pain management: American Pain Society Quality of Care Task Force. AB - BACKGROUND: The American Pain Society (APS) set out to revise and expand its 1995 Quality Improvement Guidelines for the Treatment of Acute Pain and Cancer Pain and to facilitate improvements in the quality of pain management in all care settings. METHODS: Eleven multidisciplinary members of the APS with expertise in quality improvement or measurement participated in the update. Five experts from organizations that focus on health care quality reviewed the final recommendations. MEDLINE and Cumulative Index to Nursing and Allied Health Literature databases were searched (1994-2004) to identify articles on pain quality measurement and quality improvement published after the development of the 1995 guidelines. The APS task force revised and expanded recommendations on the basis of the systematic review of published studies. The more than 3000 members of the APS were invited to provide input, and the 5 experts provided additional comments. The task force synthesized reviewers' comments into the final set of recommendations. RESULTS: The recommendations specify that all care settings formulate structured, multilevel systems approaches (sensitive to the type of pain, population served, and setting of care) that ensure prompt recognition and treatment of pain, involvement of patients and families in the pain management plan, improved treatment patterns, regular reassessment and adjustment of the pain management plan as needed, and measurement of processes and outcomes of pain management. CONCLUSION: Efforts to improve the quality of pain management must move beyond assessment and communication of pain to implementation and evaluation of improvements in pain treatment that are timely, safe, evidence based, and multimodal. PMID- 16043675 TI - Safety and efficacy of a testosterone patch for the treatment of hypoactive sexual desire disorder in surgically menopausal women: a randomized, placebo controlled trial. AB - BACKGROUND: Oophorectomy reduces serum testosterone levels. We studied the efficacy and safety of transdermal testosterone in treating hypoactive sexual desire disorder in surgically menopausal women. METHODS: A 24-week, randomized, double-blind, placebo-controlled, parallel-group, multicenter trial was conducted in women (aged 24-70 years) who developed distressful low sexual desire after bilateral salpingo-oophorectomy and hysterectomy and who were receiving oral estrogen therapy. Women were randomized to receive placebo (n = 119) or testosterone patches in dosages of 150 microg/d (n = 107), 300 microg/d (n = 110), or 450 microg/d (n = 111) twice weekly for 24 weeks. Sexual desire and frequency of satisfying sexual activity were primary efficacy outcome measures. RESULTS: Of the 447 women randomized, 318 (71%) completed the trial. Compared with placebo, women receiving the 300-microg/d testosterone patch had significantly greater increases from baseline in sexual desire (67% vs 48%; P = .05) and in frequency of satisfying sexual activity (79% vs 43%; P = .049). The 150-microg/d group showed no evidence of a treatment effect. The 450-microg/d group also was not statistically different from the 300-microg/d or placebo groups. Marginally significant linear dose-response trends were observed for total satisfying sexual activity and sexual desire at 24 weeks (P = .06 and .06, respectively). Adverse events occurred with similar frequency in both groups; no serious safety concerns were observed. CONCLUSIONS: The 300-microg/d testosterone patch increased sexual desire and frequency of satisfying sexual activity and was well tolerated in women who developed hypoactive sexual desire disorder after surgical menopause. PMID- 16043676 TI - Breast biopsy utilization: a population-based study. AB - BACKGROUND: Breast biopsy, to determine the nature of a clinical or radiographic breast abnormality, was presumed to have increased in frequency with the widespread use of screening mammography. However, scant data exist regarding the utilization of breast biopsies in the community population. METHODS: Through the resources of the Rochester Epidemiology Project, the medical records of women 18 years and older who had a breast biopsy from January 1, 1988, through December 31, 1999, were reviewed for the type of biopsy, presentation at biopsy, and tissue pathological findings. The overall and age-specific utilization rates of breast biopsies were assessed, as were changes in the breast biopsy technique after the introduction of image-guided core-needle biopsy in 1992. RESULTS: The overall annual utilization rate of breast biopsies was 62.6 per 10 000 women per year and remained stable throughout the study. Excisional breast biopsies showed a decreasing trend and core-needle biopsies increased during the study duration. The age-adjusted incidence of benign results of breast biopsies for the study duration was 38.9 per 10 000 women. The benign-malignant ratio remained constant despite changes in the biopsy procedure. CONCLUSIONS: This population-based study provides much-needed data regarding the frequency of breast biopsies and benign results of breast biopsies in a community population. The utilization rate of breast biopsies remained fairly constant throughout the study period despite the introduction of the image-guided, core-needle biopsy procedure in 1992. A multidisciplinary breast practice, along with established guidelines for breast biopsy, can ensure appropriate use of new technology and thereby improve patient care. PMID- 16043677 TI - Use of oral topiramate to promote smoking abstinence among alcohol-dependent smokers: a randomized controlled trial. AB - BACKGROUND: Previously, our group has shown that topiramate, a sulfamate substituted fructopyranose derivative, is an effective treatment for alcohol dependence. Herein, we extend that proof-of-concept study by determining whether cigarette-smoking, alcohol-dependent individuals from the earlier study also experienced improved smoking outcomes. METHODS: As a subgroup analysis of a larger double-blind, randomized, controlled, 12-week study comparing topiramate vs placebo as treatment for alcohol dependence, a 12-week clinical trial compared topiramate vs placebo in 94 cigarette-smoking, alcohol-dependent individuals. Of these, 45 were assigned to receive topiramate (escalating dose from 25 to 300 mg/d) and the remaining 49 had placebo as an adjunct to weekly standardized medication compliance management. The primary outcome was smoking cessation ascertained by self-report and confirmed by the level of serum cotinine (nicotine's major metabolite). RESULTS: Topiramate recipients were significantly more likely than placebo recipients to abstain from smoking (odds ratio, 4.46; 95% confidence interval, 1.08-18.39; P = .04). Using a serum cotinine level of 28 ng/mL or lower to segregate nonsmokers from smokers, we found that the topiramate group had 4.97 times the odds of being nonsmokers (95% confidence interval, 1.1 23.4;P = .04). Smoking cessation rates for topiramate recipients were 19.4% and 16.7% at weeks 9 and 12, respectively, compared with 6.9% at both time points for placebo recipients. CONCLUSION: In this trial, topiramate (up to 300 mg/d) showed potential as a safe and promising medication for the treatment of cigarette smoking in alcohol-dependent individuals. PMID- 16043678 TI - Smoking and other lifestyle factors and the risk of Graves' hyperthyroidism. AB - BACKGROUND: Hyperthyroidism caused by Graves' disease is common in women, yet little is known about risk factors for the disease. We sought to determine whether lifestyle factors, including smoking, alcohol consumption, physical activity level, and body mass index, are risk factors for Graves' hyperthyroidism. METHODS: This analysis was conducted using data from the Nurses' Health Study II, among 115109 women aged 25 to 42 at entry. Incident reports of women with Graves' hyperthyroidism, confirmed to have the disorder, were included. RESULTS: During 1 328 270 person-years of follow-up, incident diagnoses of Graves' hyperthyroidism were confirmed in 543 women; the 12-year incidence was 4.6 per 1000 women. Cigarette smoking was a predictor of Graves' hyperthyroidism. The hazard ratio among current smokers was 1.93 (95% confidence interval [CI], 1.54-2.43), and among past smokers it was 1.27 (95% CI, 1.03-1.56), after adjusting for recent pregnancy, parity, and other variables. Among current smokers, the hazard ratio increased with the intensity of smoking and was 2.63 (95% CI, 1.71-4.04) among women who smoked 25 or more cigarettes daily. Obesity was associated with a decreased risk of Graves' hyperthyroidism. The hazard ratio for the disorder among women with a body mass index of 30 kg/m(2) or higher was 0.68 (95% CI, 0.49-0.92). Alcohol intake and physical activity level were not associated with risk of Graves' hyperthyroidism. CONCLUSIONS: Smoking is a risk factor for Graves' hyperthyroidism in women. Obesity may be associated with a reduced risk, although weight loss as the first manifestation of hyperthyroidism cannot be excluded. PMID- 16043679 TI - Nontraumatic fracture risk with diabetes mellitus and impaired fasting glucose in older white and black adults: the health, aging, and body composition study. AB - BACKGROUND: Diabetes mellitus (DM) and related complications may increase clinical fracture risk in older adults. METHODS: Our objectives were to determine if type 2 diabetes mellitus or impaired fasting glucose was associated with higher fracture rates in older adults and to evaluate how diabetic individuals with fractures differed from those without fractures. The Health, Aging, and Body Composition Study participants were well-functioning, community-dwelling men and women aged 70 to 79 years (N = 2979; 42% black), of whom 19% had DM and 6% had impaired fasting glucose at baseline. Incident nontraumatic clinical fractures were verified by radiology reports for a mean +/- SD of 4.5 +/- 1.1 years. Cox proportional hazards regression models determined how DM and impaired fasting glucose affected subsequent risk of fracture. RESULTS: Diabetes mellitus was associated with elevated fracture risk (relative risk, 1.64; 95% confidence interval, 1.07-2.51) after adjustment for a hip bone mineral density (BMD) and fracture risk factors. Impaired fasting glucose was not significantly associated with fractures (relative risk, 1.34; 95% confidence interval, 0.67-2.67). Diabetic participants with fractures had lower hip BMD (0.818 g/cm(2) vs 0.967 g/cm(2); P<.001) and lean mass (44.3 kg vs 51.7 kg) and were more likely to have reduced peripheral sensation (35% vs 14%), transient ischemic attack/stroke (20% vs 8%), a lower physical performance battery score (5.0 vs 7.0), and falls (37% vs 21%) compared with diabetic participants without fractures (P<.05). CONCLUSIONS: These results indicate that older white and black adults with DM are at higher fracture risk compared with nondiabetic adults with a similar BMD since a higher risk of nontraumatic fractures was found after adjustment for hip BMD. Fracture prevention needs to target specific risk factors found in older adults with DM. PMID- 16043680 TI - A randomized controlled trial of vitamin D3 supplementation in African American women. AB - BACKGROUND: We conducted a randomized, placebo-controlled, double-blind trial to test the hypothesis that vitamin D(3) supplementation would prevent bone loss in calcium-replete, African American postmenopausal women. METHODS: Two hundred eight healthy black postmenopausal women, 50 to 75 years of age, were assigned to receive either placebo or 20 microg/d (800 IU) of vitamin D(3). Calcium supplements were provided to ensure a total calcium intake of 1200 to 1500 mg/d. After 2 years, the vitamin D(3) dose was increased to 50 microg/d (2000 IU) in the active group, and the study continued for an additional year. Bone mineral density (BMD) was measured every 6 months. Markers of bone turnover, vitamin D metabolites, and parathyroid hormone (PTH) levels were measured in serum. RESULTS: There were no significant differences in BMD between the active and control groups throughout the study. There was also no relationship between serum 25-hydroxyvitamin D levels attained and rates of bone loss. There was an increase in BMD of the total body, hip, and radius at 1 year in both groups. Over the 3 years, BMD declined at these sites by 0.26% to 0.55% per year. The BMD of the lumbar spine increased slightly in the placebo and active groups. There were no persistent changes in serum PTH levels or the markers of bone turnover, although there was a transient decline in PTH in both groups at 3 months. No significant adverse events were attributed to vitamin D supplementation. CONCLUSIONS: There was no observed effect of vitamin D(3) supplementation on bone loss or bone turnover markers in calcium-replete, postmenopausal African American women. Further studies are needed to determine if these findings are applicable to women of other ethnic groups. PMID- 16043681 TI - Paradoxical effect of body mass index on survival in rheumatoid arthritis: role of comorbidity and systemic inflammation. AB - BACKGROUND: Despite high cardiovascular mortality in rheumatoid arthritis (RA), few studies of body mass index (BMI) and obesity as risk factors for death in RA have been published. METHODS: We estimated the effect of BMI on survival in a cohort of 779 patients with RA adjusting for comorbidity, RA disease severity, erythrocyte sedimentation rate (ESR), and other potential confounders. RESULTS: The cohort accrued 123 deaths in 3460 person-years (3.6 deaths per 100 person years; 95% confidence interval [CI], 3.0-4.2). The BMI was inversely associated with mortality. Patients with BMIs of 30 or higher had the lowest mortality, 1.7 deaths per 100 person-years (95% CI, 1.1-2.5). Mortality was higher in each lower BMI category, reaching its highest rate among patients with BMIs lower than 20 with 15.0 deaths per 100 person-years (95% CI, 9.9-23.0). The survival advantage of high BMI was independent of RA onset age, RA duration, sex, ethnic group, socioeconomic status, smoking status, and use of methotrexate but was lost on adjusting for comorbidity and RA severity. We observed an interaction between BMI and ESR, where the BMI protective influence occurred only if the ESR was low. The BMI x ESR interaction was independent of all covariates, including comorbidity and RA severity. CONCLUSIONS: Body mass has a paradoxical effect on mortality in RA. Patients with high BMI have lower mortality than thinner patients. This effect is mediated in part by comorbidity. The effect of body mass on survival seems to be modified by the level of systemic inflammation. PMID- 16043682 TI - Quality of care by classification of myocardial infarction: treatment patterns for ST-segment elevation vs non-ST-segment elevation myocardial infarction. AB - BACKGROUND: Practice guidelines for acute ST-segment elevation myocardial infarction (STEMI) and non-STEMI (NSTEMI) recommend similar therapies and interventions, but differences in patterns of care between MI categories have not been well described in contemporary practice. METHODS: In-hospital treatments with similar recommendations from practice guidelines were compared with outcomes in 185 968 eligible patients (without listed contraindications) with STEMI (n = 53 417; 29%) vs NSTEMI (n = 132 551; 71%) from 1247 US hospitals participating in the National Registry of Myocardial Infarction 4 between July 1, 2000, and June 30, 2002. Hierarchical logistic regression modeling was used to determine adjusted differences in treatment patterns in MI categories. RESULTS: Unadjusted in-hospital mortality rates were high for NSTEMI (12.5%) and STEMI (14.3%), and the use of guideline-recommended medications and interventions was suboptimal in both categories of patients with MI. The adjusted likelihood of receiving early (within 24 hours of presentation) aspirin, beta-blockers, and angiotensin converting enzyme inhibitors was higher in patients with STEMI. Similar patterns of care were noted at hospital discharge: the adjusted likelihood of receiving aspirin, beta-blockers, angiotensin-converting enzyme inhibitors, lipid-lowering agents, smoking cessation counseling, and cardiac rehabilitation referral was higher in patients with STEMI. CONCLUSIONS: Evidence-based medications and lifestyle modification interventions were used less frequently in patients with NSTEMI. Quality improvement interventions designed to narrow the gaps in care between NSTEMI and STEMI and to improve adherence to guidelines for both categories of patients with MI may reduce the high mortality rates associated with acute MI in contemporary practice. PMID- 16043683 TI - Incidence and characteristics of angioedema associated with enalapril. AB - BACKGROUND: Angioedema is a rare but potentially serious adverse event of angiotensin-converting enzyme inhibitor therapy. However, no prospective, controlled studies have reported on its incidence and clinical characteristics. METHODS: We studied the occurrence of angioedema in a randomized, double-blind, controlled trial of 12 557 persons with hypertension treated with enalapril maleate, 5 to 40 mg/d, using a prospective ascertainment and adjudication of angioedema by an expert committee. RESULTS: Angioedema occurred in 86 (0.68%) of the subjects. Stepwise logistic regression identified black race (odds ratio [OR], 2.88; 95% confidence interval [CI], 1.72-4.82), history of drug rash (OR, 3.78; 95% CI, 1.80-7.92), age greater than 65 years (OR, 1.60; 95% CI, 1.02 2.53), and seasonal allergies (OR, 1.79; 95% CI, 1.06-3.00) as independent risk factors for angioedema. The incidence of angioedema was higher after initiation of therapy (3.6/1000 patients per month) and declined to 0.4/1000 patients per month. Treatment was not given in 44 (51%) of the cases; antihistamines were administered in 35 (41%); corticosteroids, in 20 (23%); and epinephrine, in 1 (1%). Two patients were hospitalized but none had airway compromise. CONCLUSIONS: Enalapril-related angioedema is uncommon. Although it is most likely to occur early after initiation of therapy, it may occur at any time. It is more likely to occur in black patients, those older than 65 years, and those with a history of drug rash or seasonal allergies. Fatal angioedema or angioedema requiring airway protection did not occur in this study. PMID- 16043684 TI - Systemic inflammatory response syndrome after acute myocardial infarction complicated by cardiogenic shock. AB - BACKGROUND: The role of inflammation in patients with coronary artery disease is emerging. We sought to assess the profile and outcomes of patients with a clinical syndrome of severe systemic inflammation that led to a diagnosis of suspected sepsis in the setting of acute myocardial infarction complicated by cardiogenic shock (CS). METHODS: Patients enrolled in the randomized SHOCK (SHould we emergently revascularize Occluded Coronaries for cardiogenic shocK) trial (n = 302) were divided into those with clinical signs of severe systemic inflammation (eg, fever [94%] or leukocytosis [72%]) that led to a diagnosis of suspected sepsis (n = 54 [18%]) and those without suspected sepsis (controls; n = 243 [80%]). The patients with suspected sepsis were then further subdivided into those who were considered to be potentially infectious (positive culture result ["culture-positive"]; n = 40) and those who were not (negative culture result ["culture-negative"]; n = 14). RESULTS: Severe systemic inflammation was diagnosed 4 and 2 days after the onset of CS in culture-positive and culture negative patients, respectively. Patients who developed systemic inflammation tended to be younger (P = .05) and to have lower systemic vascular resistance (SVR) near the onset of CS (P = .006). Many culture-positive patients (40%) had undergone coronary artery bypass graft surgery. However, the lower the initial SVR, the higher the risk of developing culture-positive systemic inflammation (P = .01), even after controlling for age and coronary artery bypass graft surgery. A time-dependent model, adjusted for age, showed that culture-positive patients were at significantly higher risk for death than were controls (hazard ratio, 2.22; 95% confidence interval, 1.32-3.76; P = .008). CONCLUSIONS: Almost one fifth of patients with acute myocardial infarction complicated by CS showed clinical signs of severe systemic inflammation, and those who were culture positive for sepsis had twice the risk of death. The observation of lower SVR at the onset of shock in patients who subsequently had culture-positive systemic inflammation suggests that inappropriate vasodilation may play an important role in the pathogenesis and persistence of shock and in the risk of infection. PMID- 16043685 TI - Plasma C peptide level and cognitive function among older women without diabetes mellitus. AB - BACKGROUND: Growing evidence suggests that type 2 diabetes mellitus and hyperinsulinemia may be related to diminished cognition. To help differentiate between the effects of diabetes and insulin, we examined the relation of insulin to cognitive function among nondiabetic participants of the Nurses' Health Study. METHODS: We measured the C peptide level, representing insulin secretion, in blood samples provided by 718 women from June 14, 1989, to October 4, 1990, when they were aged 61 to 69 years. We administered telephone interviews an average of 10 years after blood collection, testing general cognition, verbal memory, category fluency, and attention; second cognitive assessments were conducted 2 years later. The primary outcomes were global cognitive function across all tests and verbal memory. We used regression models to estimate multivariable-adjusted mean differences in cognitive function and cognitive decline, and odds of cognitive impairment, across C peptide levels. RESULTS: Cognitive function was worse among women in the fourth C peptide quartile compared with those in the first quartile (eg, on the global score combining all cognitive tests, the multivariable-adjusted mean difference was -1.7 standard units [95% confidence interval, -2.9 to -0.6 standard units]; P = .002); the odds of cognitive impairment (defined as the worst 10% of the distribution) were 3-fold higher among women in the fourth vs first quartile (95% confidence interval, 1.3-7.8). On verbal memory, women in the fourth quartile scored significantly worse than those in the first quartile; the odds of impairment were 2.8-fold higher (95% confidence interval, 1.1-7.0). Consistent findings were observed for cognitive decline. CONCLUSION: Higher insulin secretion may be related to worse cognition, even among those without diabetes. PMID- 16043686 TI - Premature death associated with delirium at 1-year follow-up. AB - BACKGROUND: While previous studies have demonstrated the increased mortality risk associated with delirium, little is known about the mortality time course. The objective of this study is to estimate the fraction of a year of life lost associated with delirium at 1-year follow-up. METHODS: Hospitalized patients 70 years and older who participated in a previous controlled clinical trial of a delirium prevention intervention at an academic medical center from March 25, 1995, through March 18, 1998, were followed up for 1 year after discharge, and patients who died were identified, along with the date of death. The adjusted number of days survived were estimated using a 2-step regression model approach and compared across patients who developed delirium during hospitalization and those who did not develop delirium. RESULTS: After adjusting for pertinent covariates (age, sex, functional status, and comorbidity), patients with delirium survived 274 days, compared with 321 days for patients without delirium, representing a difference of 13% of a year (hazard ratio, 1.62; P<.001). Results were confirmed with a separate binomial regression analysis. CONCLUSIONS: Patients who experienced delirium during hospitalization had a 62% increased risk of mortality and lost an average of 13% of a year of life compared with patients without delirium. Although delirium is an acute condition, it is associated with multiple long-term sequelae that extend beyond the hospital setting, including premature mortality. PMID- 16043687 TI - Dilated cardiomyopathy and Creutzfeldt-Jakob disease: evidence for a role of cellular prion protein in the heart? PMID- 16043688 TI - Vasopressin has cost and administration advantages over epinephrine in cardiac arrest. PMID- 16043689 TI - Controlling for patient risk in volume-outcome studies. PMID- 16043691 TI - Comparison of the complete genome sequences of Pseudomonas syringae pv. syringae B728a and pv. tomato DC3000. AB - The complete genomic sequence of Pseudomonas syringae pv. syringae B728a (Pss B728a) has been determined and is compared with that of P. syringae pv. tomato DC3000 (Pst DC3000). The two pathovars of this economically important species of plant pathogenic bacteria differ in host range and other interactions with plants, with Pss having a more pronounced epiphytic stage of growth and higher abiotic stress tolerance and Pst DC3000 having a more pronounced apoplastic growth habitat. The Pss B728a genome (6.1 Mb) contains a circular chromosome and no plasmid, whereas the Pst DC3000 genome is 6.5 mbp in size, composed of a circular chromosome and two plasmids. Although a high degree of similarity exists between the two sequenced Pseudomonads, 976 protein-encoding genes are unique to Pss B728a when compared with Pst DC3000, including large genomic islands likely to contribute to virulence and host specificity. Over 375 repetitive extragenic palindromic sequences unique to Pss B728a when compared with Pst DC3000 are widely distributed throughout the chromosome except in 14 genomic islands, which generally had lower GC content than the genome as a whole. Content of the genomic islands varies, with one containing a prophage and another the plasmid pKLC102 of Pseudomonas aeruginosa PAO1. Among the 976 genes of Pss B728a with no counterpart in Pst DC3000 are those encoding for syringopeptin, syringomycin, indole acetic acid biosynthesis, arginine degradation, and production of ice nuclei. The genomic comparison suggests that several unique genes for Pss B728a such as ectoine synthase, DNA repair, and antibiotic production may contribute to the epiphytic fitness and stress tolerance of this organism. PMID- 16043692 TI - Genome-wide expression profiling of human blood reveals biomarkers for Huntington's disease. AB - Huntington's disease (HD) is an autosomal dominant disorder caused by an expansion of glutamine repeats in ubiquitously distributed huntingtin protein. Recent studies have shown that mutant huntingtin interferes with the function of widely expressed transcription factors, suggesting that gene expression may be altered in a variety of tissues in HD, including peripheral blood. Affymetrix and Amersham Biosciences oligonucleotide microarrays were used to analyze global gene expression in blood samples of HD patients and matched controls. We identified 322 mRNAs that showed significantly altered expression in HD blood samples, compared with controls (P < 0.0005), on two different microarray platforms. A subset of up-regulated mRNAs selected from this group was able to distinguish controls, presymptomatic individuals carrying the HD mutation, and symptomatic HD patients. In addition, early presymptomatic subjects showed gene expression profiles similar to those of controls, whereas late presymptomatic subjects showed altered expression that resembled that of symptomatic HD patients. These elevated mRNAs were significantly reduced in HD patients involved in a dose finding study of the histone deacetylase inhibitor sodium phenylbutyrate. Furthermore, expression of the marker genes was significantly up-regulated in postmortem HD caudate, suggesting that alterations in blood mRNAs may reflect disease mechanisms observed in HD brain. In conclusion, we identified changes in blood mRNAs that clearly distinguish HD patients from controls. These alterations in mRNA expression correlate with disease progression and response to experimental treatment. Such markers may provide clues to the state of HD and may be of predictive value in clinical trials. PMID- 16043693 TI - The structure of phospholamban pentamer reveals a channel-like architecture in membranes. AB - Contraction and relaxation of heart muscle cells is regulated by cycling of calcium between cytoplasm and sarcoplasmic reticulum. Human phospholamban (PLN), expressed in the sarcoplasmic reticulum membrane as a 30-kDa homopentamer, controls cellular calcium levels by a mechanism that depends on its phosphorylation. Since PLN was discovered approximately 30 years ago, extensive studies have aimed to explain how it influences calcium pumps and to determine whether it acts as an ion channel. We have determined by solution NMR methods the atomic resolution structure of an unphosphorylated PLN pentamer in dodecylphosphocholine micelles. The unusual bellflower-like assembly is held together by leucine/isoleucine zipper motifs along the membrane-spanning helices. The structure reveals a channel-forming architecture that could allow passage of small ions. The central pore gradually widens toward the cytoplasmic end as the transmembrane helices twist around each other and bend outward. The dynamic N terminal amphipathic helices point away from the membrane, perhaps facilitating recognition and inhibition of the calcium pump. PMID- 16043694 TI - Magnetic isotope effect of magnesium in phosphoglycerate kinase phosphorylation. AB - Phosphoglycerate kinase (PGK) is found to be controlled by a (25)Mg(2+)-related magnetic isotope effect. Mg(2+) nuclear spin selectivity manifests itself in PGK directed ADP phosphorylation, which has been clearly proven by comparison of ATP synthesis rates estimated in reaction mixtures with different Mg isotopy parameters. Both pure (25)Mg(2+) (nuclear spin 5/2, magnetic moment +0.85) and (24)Mg(2+) (spinless, nonmagnetic nucleus) species as well as their mixtures were used in experiments. In the presence of (25)Mg(2+), ATP production is 2.6 times higher compared with the yield of ATP reached in (24)Mg(2+)-containing PGK-based catalytic systems. The chemical mechanism of this phenomenon is discussed. A key element of the mechanism proposed is a nonradical pair formation in which (25)Mg(+) radical cation and phosphate oxyradical are involved. PMID- 16043695 TI - HIV-1 Nef down-regulates the hemochromatosis protein HFE, manipulating cellular iron homeostasis. AB - The multifunctional Nef protein of HIV-1 is important for the progression to AIDS. One action of Nef is to down-regulate surface MHC I molecules, helping infected cells to evade immunity. We found that Nef also down-regulates the macrophage-expressed MHC 1b protein HFE, which regulates iron homeostasis and is mutated in the iron-overloading disorder hemochromatosis. In model cell lines, Nef reroutes HFE to a perinuclear structure that overlaps the trans-Golgi network, causing a 90% reduction of surface HFE. This activity requires a Src kinase-binding proline-rich domain of Nef and a conserved tyrosine-based motif in the cytoplasmic tail of HFE. HIV-1 infection of ex vivo macrophages similarly down-regulates naturally expressed surface HFE in a Nef-dependent manner. The effect of Nef expression on cellular iron was explored; iron and ferritin accumulation were increased in HIV-1-infected ex vivo macrophages expressing wild type HFE, but this effect was lost with Nef-deleted HIV-1 or when infecting macrophages from hemochromatosis patients expressing mutated HFE. The iron accumulation in HIV-1-infected HFE-expressing macrophages was paralleled by an increase in cellular HIV-1-gag expression. We conclude that, through Nef and HFE, HIV-1 directly regulates cellular iron metabolism, possibly benefiting viral growth. PMID- 16043696 TI - Cohesins are required for meiotic DNA breakage and recombination in Schizosaccharomyces pombe. AB - In preparation for the unique segregation of homologs at the first meiotic division, chromosomes undergo dramatic changes. The meiosis-specific sister chromatid cohesins Rec8 and Rec11 of Schizosaccharomyces pombe are recruited around the time of premeiotic replication, and Rec10, a component of meiosis specific linear elements, is subsequently added. Here we report that Rec10 is essential for meiosis-specific DNA breakage by Rec12 (Spo11 homolog) and for meiotic recombination. DNA breakage and recombination also depend on the Rec8 and Rec11 cohesins, strictly in some genomic intervals but less so in others. Thus, in addition to their previously recognized role in meiotic chromosome segregation, cohesins have a direct role, as do linear element components, in meiotic recombination by enabling double-strand DNA break formation by Rec12. Our results reveal a pathway, whose regulation is significantly different from that in the distantly related yeast Saccharomyces cerevisiae, for meiosis-specific chromosome differentiation and high-frequency recombination. PMID- 16043697 TI - Iron and Pseudomonas aeruginosa biofilm formation. AB - Iron serves as a signal in Pseudomonas aeruginosa biofilm development. We examined the influence of mutations in known and putative iron acquisition signaling genes on biofilm morphology. In iron-sufficient medium, mutants that cannot obtain iron through the high-affinity pyoverdine iron acquisition system form thin biofilms similar to those formed by the parent under low iron conditions. If an iron source for a different iron acquisition system is provided to a pyoverdine mutant, normal biofilm development occurs. This enabled us to identify iron uptake gene clusters that likely serve in transport of ferric citrate and ferrioxamine. We suggest that the functional iron signal for P. aeruginosa biofilm development is active transport of chelated iron or the level of internal iron. If the signal is internal iron levels, then a factor likely to be involved in iron signaling is the cytoplasmic ferric uptake regulator protein, Fur, which controls expression of iron-responsive genes. In support of a Fur involvement, we found that with low iron a Fur mutant was able to organize into more mature biofilms than was the parent. The two known Fur-controlled small regulatory RNAs (PrrF1 and F2) do not appear to mediate iron control of biofilm development. This information establishes a mechanistic basis for iron control of P. aeruginosa biofilm formation. PMID- 16043698 TI - Electrically monitoring DNA repair by photolyase. AB - Cyclobutane pyrimidine dimers are the major DNA photoproducts produced upon exposure to UV radiation. If left unrepaired, these lesions can lead to replication errors, mutation, and cell death. Photolyase is a light-activated flavoenzyme that binds to pyrimidine dimers in DNA and repairs them in a reaction triggered by electron transfer from the photoexcited flavin cofactor to the dimer. Using gold electrodes modified with DNA duplexes containing a cyclobutane thymine dimer (T<>T), here we probe the electrochemistry of the flavin cofactor in Escherichia coli photolyase. Cyclic and square-wave voltammograms of photolyase deposited on these electrodes show a redox signal at 40 mV versus normal hydrogen electrode, consistent with electron transfer to and from the flavin in the DNA-bound protein. This signal is dramatically attenuated on surfaces where the pi-stacking of the DNA bases is perturbed by the presence of an abasic site below the T<>T, an indication that the redox pathway is DNA mediated. DNA repair can, moreover, be monitored electrically. Exposure of photolyase on T<>T-damaged DNA films to near-UV/blue light leads to changes in the flavin signal consistent with repair, as confirmed by parallel HPLC experiments. These results demonstrate the exquisite sensitivity of DNA electrochemistry to perturbations in base pair stacking and the applicability of this chemistry to probe reactions of proteins with DNA. PMID- 16043699 TI - Noise-induced stability in dryland plant ecosystems. AB - Dryland plant ecosystems tend to exhibit bistable dynamics with two preferential configurations of bare and vegetated soils. Climate fluctuations are usually believed to act as a source of disturbance on these ecosystems and to reduce their stability and resilience. In contrast, this work shows that random interannual fluctuations of precipitation may lead to the emergence of an intermediate statistically stable condition between the two stable states of the deterministic dynamics of vegetation. As a result, there is an enhancement of ecosystem resilience and a decrease in the likelihood of catastrophic shifts to the desert state. PMID- 16043700 TI - Structure, function, and evolution of transient and obligate protein-protein interactions. AB - Recent analyses of high-throughput protein interaction data coupled with large scale investigations of evolutionary properties of interaction networks have left some unanswered questions. To what extent do protein interactions act as constraints during evolution of the protein sequence? How does the type of interaction, specifically transient or obligate, play into these constraints? Are the mutations in the binding site of an interacting protein correlated with mutations in the binding site of its partner? We address these and other questions by relying on a carefully curated dataset of protein complex structures. Results point to the importance of distinguishing between transient and obligate interactions. We conclude that residues in the interfaces of obligate complexes tend to evolve at a relatively slower rate, allowing them to coevolve with their interacting partners. In contrast, the plasticity inherent in transient interactions leads to an increased rate of substitution for the interface residues and leaves little or no evidence of correlated mutations across the interface. PMID- 16043701 TI - Possible role of the 11-cis-retinyl conformation in controlling the dual decay processes of excited rhodopsin. AB - In this work, we examine how the reported dual decay processes of rhodopsin and binding site stereospecificity can be accounted for by the recently available crystal structure of rhodopsin. Arguments are presented for possible presence of two rhodopsin "rotamers" that fit within the binding cavity. Directed pathways of decay could account for the observed excited decay processes. We summarize evidence for the possible existence of two different ground-state configurations that give rise to two different excited species. PMID- 16043702 TI - Drier summers cancel out the CO2 uptake enhancement induced by warmer springs. AB - An increase in photosynthetic activity of the northern hemisphere terrestrial vegetation, as derived from satellite observations, has been reported in previous studies. The amplitude of the seasonal cycle of the annually detrended atmospheric CO(2) in the northern hemisphere (an indicator of biospheric activity) also increased during that period. We found, by analyzing the annually detrended CO(2) record by season, that early summer (June) CO(2) concentrations indeed decreased from 1985 to 1991, and they have continued to decrease from 1994 up to 2002. This decrease indicates accelerating springtime net CO(2) uptake. However, the CO(2) minimum concentration in late summer (an indicator of net growing-season uptake) showed no positive trend since 1994, indicating that lower net CO(2) uptake during summer cancelled out the enhanced uptake during spring. Using a recent satellite normalized difference vegetation index data set and climate data, we show that this lower summer uptake is probably the result of hotter and drier summers in both mid and high latitudes, demonstrating that a warming climate does not necessarily lead to higher CO(2) growing-season uptake, even in high-latitude ecosystems that are considered to be temperature limited. PMID- 16043703 TI - Investigating diversity in human plasma proteins. AB - Plasma proteins represent an important part of the human proteome. Although recent proteomics research efforts focus largely on determining the overall number of proteins circulating in plasma, it is equally important to delineate protein variations among individuals, because they can signal the onset of diseases and be used as biological markers in diagnostics. To date, there has been no systematic proteomics effort to characterize the breadth of structural modifications in individual proteins in the general population. In this work, we have undertaken a population proteomics study to define gene- and protein-level diversity that is encountered in the general population. Twenty-five plasma proteins from a cohort of 96 healthy individuals were investigated through affinity-based mass spectrometric assays. A total of 76 structural forms/variants were observed for the 25 proteins within the samples cohort. Posttranslational modifications were detected in 18 proteins, and point mutations were observed in 4 proteins. The frequency of occurrence of these variations was wide-ranged, with some modifications being observed in only one sample, and others detected in all 96 samples. Even though a relatively small cohort of individuals was investigated, the results from this study illustrate the extent of protein diversity in the human population and can be of immediate aid in clinical proteomics/biomarker studies by laying a basal-level statistical foundation from which protein diversity relating to disease can be evaluated. PMID- 16043704 TI - The molecular structure of the Toll-like receptor 3 ligand-binding domain. AB - Innate immunity is the first line of defense against invading pathogens. Toll like receptors (TLRs) act as sentinels of the innate immune system, sensing a variety of ligands from lipopolysaccharide to flagellin to dsRNA through their ligand-binding domain that is composed of leucine-rich repeats (LRRs). Ligand binding initiates a signaling cascade that leads to the up-regulation of inflammation mediators. In this study, we have expressed and crystallized the ectodomain (ECD) of human TLR3, which recognizes dsRNA, a molecular signature of viruses, and have determined the molecular structure to 2.4-A resolution. The overall horseshoe-shaped structure of the TLR3-ECD is formed by 23 repeating LRRs that are capped at each end by specialized non-LRR domains. The extensive beta sheet on the molecule's concave surface forms a platform for several modifications, including insertions in the LRRs and 11 N-linked glycans. The TLR3 ECD structure indicates how LRR loops can establish distinct pathogen recognition receptors. PMID- 16043705 TI - Mid- to late-Holocene El Nino-Southern Oscillation dynamics reflected in the subtropical terrestrial realm. AB - High resolution pollen analysis of mid- to late-Holocene peat deposits from southwest Florida reveals a stepwise increase in wetland vegetation that points to an increased precipitation-driven fresh water flow during the past 5,000 years. The tight coupling between winter precipitation patterns in Florida and the strength of the El Nino-Southern Oscillation (ENSO) strongly suggests that the paleo-hydrology record reflects changes in ENSO intensity. A terrestrial subtropical record outside the Indo Pacific Warm Pool both documents ecosystem response to the known onset of modern-day ENSO periodicities, between approximately 7,000 and 5,000 years B.P., and subsequent ENSO intensification after 3,500 years B.P. The observed increases in "wetness" are sustained by a gradual rise in relative sea level that prevents a return to drier vegetation through natural succession. PMID- 16043706 TI - Space-time thermodynamics of the glass transition. AB - We consider the probability distribution for fluctuations in dynamical action and similar quantities related to dynamic heterogeneity. We argue that the so-called "glass transition" is a manifestation of low action tails in these distributions where the entropy of trajectory space is subextensive in time. These low action tails are a consequence of dynamic heterogeneity and an indication of phase coexistence in trajectory space. The glass transition, where the system falls out of equilibrium, is then an order-disorder phenomenon in space-time occurring at a temperature T(g), which is a weak function of measurement time. We illustrate our perspective ideas with facilitated lattice models and note how these ideas apply more generally. PMID- 16043707 TI - Automated projection spectroscopy (APSY). AB - This work presents the automated projection spectroscopy (APSY) method for the recording of discrete sets of j projections from N-dimensional (N > or = 3) NMR experiments at operator-selected projection angles and automatic identification of the correlation cross peaks. The result from APSY is the fully automated generation of the complete or nearly complete peak list for the N-dimensional NMR spectrum from a geometric analysis of the j experimentally recorded, low dimensional projections. In the present implementation of APSY, two-dimensional projections of the N-dimensional spectrum are recorded by using techniques developed for projection-reconstruction spectroscopy [Kupce,E.& Freeman, R. (2004) J. Am. Chem. Soc. 126, 6429-6440]. All projections are peak-picked with the available automated routine atnos. The previously undescribed algorithm GAPRO (geometric analysis of projections) uses vector algebra to identify subgroups of peaks in different projections that arise from the same resonance in the N dimensional spectrum, and from these subgroups it calculates the peak positions in the N-dimensional frequency space. Unambiguous identification thus can be achieved for all cross peaks that are not overlapped with other peaks in at least one of the N dimensions. Because of the correlation between the positions of corresponding peaks in multiple projections, uncorrelated noise is efficiently suppressed, so that APSY should be quite widely applicable for correlation spectra of biological macromolecules, which have intrinsically low peak density in the N-dimensional spectral space. PMID- 16043708 TI - Agrin promotes synaptic differentiation by counteracting an inhibitory effect of neurotransmitter. AB - Synaptic organizing molecules and neurotransmission regulate synapse development. Here, we use the skeletal neuromuscular junction to assess the interdependence of effects evoked by an essential synaptic organizing protein, agrin, and the neuromuscular transmitter, acetylcholine (ACh). Mice lacking agrin fail to maintain neuromuscular junctions, whereas neuromuscular synapses differentiate extensively in the absence of ACh. We now demonstrate that agrin's action in vivo depends critically on cholinergic neurotransmission. Using double-mutant mice, we show that synapses do form in the absence of agrin provided that ACh is also absent. We provide evidence that ACh destabilizes nascent postsynaptic sites, and that one major physiological role of agrin is to counteract this "antisynaptogenic" influence. Similar interactions between neurotransmitters and synaptic organizing molecules may operate at synapses in the central nervous system. PMID- 16043709 TI - The psychrophilic lifestyle as revealed by the genome sequence of Colwellia psychrerythraea 34H through genomic and proteomic analyses. AB - The completion of the 5,373,180-bp genome sequence of the marine psychrophilic bacterium Colwellia psychrerythraea 34H, a model for the study of life in permanently cold environments, reveals capabilities important to carbon and nutrient cycling, bioremediation, production of secondary metabolites, and cold adapted enzymes. From a genomic perspective, cold adaptation is suggested in several broad categories involving changes to the cell membrane fluidity, uptake and synthesis of compounds conferring cryotolerance, and strategies to overcome temperature-dependent barriers to carbon uptake. Modeling of three-dimensional protein homology from bacteria representing a range of optimal growth temperatures suggests changes to proteome composition that may enhance enzyme effectiveness at low temperatures. Comparative genome analyses suggest that the psychrophilic lifestyle is most likely conferred not by a unique set of genes but by a collection of synergistic changes in overall genome content and amino acid composition. PMID- 16043710 TI - Human POT1 disrupts telomeric G-quadruplexes allowing telomerase extension in vitro. AB - The POT1 (protection of telomeres 1) protein binds the ssDNA overhangs at the ends of chromosomes in diverse eukaryotes. POT1 is essential for chromosome end protection, as best demonstrated in fission yeast. In human cells, hPOT1 is also involved in telomere-length regulation. We now show that telomeric oligonucleotides, such as d[GGG(TTAGGG)(3)], which form intramolecular G quadruplexes through Hoogsteen base-pairing, serve as only marginal primers for extension by recombinant human telomerase; telomerase stalls after every nucleotide addition. Addition of hPOT1 to the reaction restores the normal processive elongation pattern seen with primers that cannot form G-quadruplexes. hPOT1 does not act catalytically but, instead, forms a stoichiometric complex with the DNA, freeing its 3' tail. An antisense oligonucleotide, which base-pairs near the 5' end of the telomeric sequence, leaving a telomerase-extendable 3' tail, duplicates the effect of hPOT1 on activation of G-quadruplex primers. Thus, hPOT1 may function simply by trapping the unfolded forms of these telomeric primers in an equilibrium population. We propose an additional role for hPOT1 in telomere maintenance: disrupting G-quadruplex structures in telomeric DNA, thereby allowing proper elongation by telomerase. PMID- 16043711 TI - Recombinant AAV2 transduction of primitive human hematopoietic stem cells capable of serial engraftment in immune-deficient mice. AB - A recombinant AAV2 (rAAV2) vector encoding antisense RNA to HIV-1 transactivating region (TAR) was evaluated for transduction of human cord blood CD34+CD38- hematopoietic stem cells (HSC) capable of serial engraftment in nonobese diabetic (NOD)/severe combined immunodeficient (SCID) mice. Results revealed long-term multilineage marking in primary and secondary recipients, and significantly, an enrichment of transduced cells in secondary hosts, indicating efficient transduction of multipotential self-renewing HSC. These results were confirmed by the persistence of rAAV marking of clonogenic progenitors in serial analyses of recipient marrow. Upon HIV-1 challenge, the macrophage progeny of transduced CD34+ cells expressed antisense RNA and exhibited sustained and significant inhibition of virus replication as compared with controls in every donor tested, without selective pressure. This study represents a clear in vivo demonstration of efficient rAAV2 transduction of human HSC. PMID- 16043712 TI - Facilitated replacement of Kupffer cells expressing a paraoxonase-1 transgene is essential for ameliorating atherosclerosis in mice. AB - Resident macrophages (i.e., Kupffer cells) are derived from hematopoietic stem cells (HSCs) and are primarily responsible for the removal from plasma of oxidized forms of low-density lipoprotein (LDL). The therapeutic potential of Kupffer cell expression of a transgene encoding paraoxonase-1 (PON1), whose plasma activity correlates with the protection from atherosclerosis, was examined in mice rendered atherosclerosis-susceptible through genetic deletion of the LDL receptor. Mice having their bone marrow engrafted with HSCs expressing the PON1 transgene (PON1-Tg) driven by a macrophage-specific promoter were injected i.v. with saline (vehicle only) or with gadolinium chloride (GdCl(3)), an agent that rapidly causes Kupffer cell apoptosis. One month later, GdCl(3)-facilitated Kupffer cell apoptosis increased the hepatic expression of transgenic PON1 mRNA by 9-fold. After 12 weeks of being fed a cholesterol-enriched atherogenic diet, mice injected with GdCl(3) exhibited 50% reductions in both aortic sinus atherosclerotic lesions (P < 0.0097) and surface lesions of the abdominal aorta (P < 0.006). In contrast, mice receiving HSCs expressing the PON1-Tg but not treated with GdCl(3) showed no protection from atherosclerosis. In addition, mice engrafted with HSCs not expressing the PON1-Tg but injected with GdCl(3) also showed no protection from atherosclerosis. These findings, showing that GdCl(3) enhanced hepatic expression of the PON1-Tg is essential for reducing atherosclerosis, indicate that Kupffer cells play an important role in atherogenesis. GdCl(3)-facilated replacement of Kupffer cells may enhance the efficacy of other HSC-based gene therapies. PMID- 16043713 TI - Repression of phase-variable cup gene expression by H-NS-like proteins in Pseudomonas aeruginosa. AB - The cupA gene cluster of Pseudomonas aeruginosa encodes components of a putative fimbrial structure that enable this opportunistic human pathogen to form biofilms on abiotic surfaces. In P. aeruginosa, cupA gene expression is repressed by MvaT, a putative transcription regulator thought to belong to the H-NS family of nucleoid-associated proteins that typically function by repressing transcription. Here, we present evidence that MvaT controls phase-variable (ON/OFF) expression of the cupA fimbrial gene cluster. Using a directed proteomic approach, we show that MvaT associates with a related protein in P. aeruginosa called MvaU. Analysis with a bacterial two-hybrid system designed to facilitate the study of protein dimerization indicates that MvaT and MvaU can form both heteromeric and homomeric complexes, and that formation of these complexes is mediated through the N-terminal regions of MvaT and MvaU, both of which are predicted to adopt a coiled-coil conformation. We show further that, like MvaT, MvaU can repress phase variable expression of the cupA gene cluster. Our findings suggest that fimbrial genes important for biofilm formation can be expressed in a phase-variable manner in P. aeruginosa, provide insight into the molecular mechanism of MvaT-dependent gene control, and lend further weight to the postulate that MvaT proteins are H NS-like in nature. PMID- 16043714 TI - The voltage-gated potassium channel Kv1.3 is highly expressed on inflammatory infiltrates in multiple sclerosis brain. AB - Multiple Sclerosis (MS) is characterized by central nervous system perivenular and parenchymal mononuclear cell infiltrates consisting of activated T cells and macrophages. We recently demonstrated that elevated expression of the voltage gated potassium channel, Kv1.3, is a functional marker of activated effector memory T (T(EM)) cells in experimental allergic encephalomyelitis and in myelin specific T cells derived from the peripheral blood of patients with MS. Herein, we show that Kv1.3 is highly expressed in postmortem MS brain inflammatory infiltrates. The expression pattern revealed not only Kv1.3(+) T cells in the perivenular infiltrate but also high expression in the parenchyma of demyelinated MS lesions and both normal appearing gray and white matter. These cells were uniformly chemokine receptor 7 negative (CCR7(-)), consistent with an effector memory phenotype. Using double-labeling immunohistochemistry and confocal microscopy, we demonstrated colocalization of Kv1.3 with CD3, CD4, CD8, and some CD68 cells. The expression patterns mirrored in vitro experiments showing polarization of Kv1.3 to the immunological synapse. Kv1.3 was expressed in low to moderate levels on CCR7(+) central memory T cells from cerebrospinal fluid, but, when these cells were stimulated in vitro, they rapidly became Kv1.3(high)/CCR7( ) T(EM), suggesting that a subset of cerebrospinal fluid cells existed in a primed state ready to become T(EM). These studies provide further rationale for the use of specific Kv1.3 antagonists in MS. PMID- 16043715 TI - Endothelial nitric oxide synthase is critical for ischemic remodeling, mural cell recruitment, and blood flow reserve. AB - The genetic loss of endothelial-derived nitric oxide synthase (eNOS) in mice impairs vascular endothelial growth factor (VEGF) and ischemia-initiated blood flow recovery resulting in critical limb ischemia. This result may occur through impaired arteriogenesis, angiogenesis, or mobilization of stem and progenitor cells. Here, we show that after ischemic challenge, eNOS knockout mice [eNOS (-/ )] have defects in arteriogenesis and functional blood flow reserve after muscle stimulation and pericyte recruitment, but no impairment in endothelial progenitor cell recruitment. More importantly, the defects in blood flow recovery, clinical manifestations of ischemia, ischemic reserve capacity, and pericyte recruitment into the growing neovasculature can be rescued by local intramuscular delivery of an adenovirus encoding a constitutively active allele of eNOS, eNOS S1179D, but not a control virus. Collectively, our data suggest that endogenous eNOS-derived NO exerts direct effects in preserving blood flow, thereby promoting arteriogenesis, angiogenesis, and mural cell recruitment to immature angiogenic sprouts. PMID- 16043717 TI - Evolutionary cycles of cooperation and defection. AB - The main obstacle for the evolution of cooperation is that natural selection favors defection in most settings. In the repeated prisoner's dilemma, two individuals interact several times, and, in each round, they have a choice between cooperation and defection. We analyze the evolutionary dynamics of three simple strategies for the repeated prisoner's dilemma: always defect (ALLD), always cooperate (ALLC), and tit-for-tat (TFT). We study mutation-selection dynamics in finite populations. Despite ALLD being the only strict Nash equilibrium, we observe evolutionary oscillations among all three strategies. The population cycles from ALLD to TFT to ALLC and back to ALLD. Most surprisingly, the time average of these oscillations can be entirely concentrated on TFT. In contrast to the classical expectation, which is informed by deterministic evolutionary game theory of infinitely large populations, stochastic evolution of finite populations need not choose the strict Nash equilibrium and can therefore favor cooperation over defection. PMID- 16043716 TI - The gene associated with trichorhinophalangeal syndrome in humans is overexpressed in breast cancer. AB - A comprehensive differential gene expression screen on a panel of 54 breast tumors and >200 normal tissue samples using DNA microarrays revealed 15 genes specifically overexpressed in breast cancer. One of the most prevalent genes found was trichorhinophalangeal syndrome type 1 (TRPS-1), a gene previously shown to be associated with three rare autosomal dominant genetic disorders known as the trichorhinophalangeal syndromes. A number of corroborating methodologies, including in situ hybridization, e-Northern analysis using ORF EST (ORESTES) and Unigene EST abundance analysis, immunoblot and immunofluorescence analysis of breast tumor cell lines, and immunohistochemistry, confirmed the microarray findings. Immunohistochemistry analysis found TRPS-1 protein expressed in >90% of early- and late-stage breast cancer, including ductal carcinoma in situ and invasive ductal, lobular, and papillary carcinomas. The TRPS-1 gene is also immunogenic with processed and presented peptides activating T cells found after vaccination of HLA-A2.1 transgenic mouse. Human T cell lines from HLA-A*0201+ female donors exhibiting TRPS-1-specific cytotoxic T lymphocyte activity could also be generated. PMID- 16043718 TI - Disabling TNF receptor signaling by induced conformational perturbation of tryptophan-107. AB - We have disabled TNF receptor (TNFR) function by inducing allosteric modulation of tryptophan-107 (W107) in the receptor. The allosteric effect operates by means of an allosteric cavity found a short distance from a previously identified loop involved in ligand binding. Occupying this cavity by small molecules leads to perturbation of distal W107 and disables functions of the TNFR, a molecule not known to undergo conformational change upon binding TNF-alpha. TNF-alpha-induced NF-kappaB and p38 kinase activities and clinical symptoms of collagen-induced arthritis in mice were all diminished. Thus, disabling receptor function by induced conformational changes of active binding surfaces represents an innovative paradigm in structure-based drug design. PMID- 16043719 TI - Permeation-driven flow in poly(dimethylsiloxane) microfluidic devices. AB - Poly(dimethylsiloxane) is currently the material of choice for rapidly fabricating microfluidic devices. As the size of these devices decreases, a significant hydrodynamic flow is generated due to permeation of fluid through the channel walls. We develop a theoretical model verified by single bead tracking experiments, which demonstrates that large flow rates (>10 microm/s) can be passively generated in a straight microchannel filled with water. Realizing that this flow may be unwanted in some applications, we present a method to eliminate it by inhibiting mass transfer of water into the poly(dimethylsiloxane) walls. Furthermore, we explore applications to harness this passively generated flow inside a microfluidic device such as bead stacking, chemical concentration, and passive pumping. PMID- 16043720 TI - Functional cavitands: chemical reactivity in structured environments. AB - Container-shaped molecules provide structured environments that impart geometric bounds on the motions and conformations of smaller molecular occupants. Moreover, they provide "solvation" that is constrained in time and space. When inwardly directed functional groups are present, they can interact chemically with the occupants. Additionally, the potential for reactivity and catalysis is greatly enhanced. Deep cavitands, derived from resorcinarenes, nearly surround smaller molecules and have been one of the most successful platforms for elaboration with functional groups. Derivatives bearing organic and metal-binding functional groups have been shown to affect recognition properties and selectively accelerate diverse reactions. In this review, we examine recent examples of these systems with an emphasis on how and why ordered nanoenvironments impart changes in the properties and reactivity of their occupants. PMID- 16043721 TI - Self-assembled graphitic nanotubes with one-handed helical arrays of a chiral amphiphilic molecular graphene. AB - Self-assembly of a Gemini-shaped, chiral amphiphilic hexa-peri-hexabenzocoronene having two chiral oxyalkylene side chains, along with two lipophilic side chains, yields graphitic nanotubes with one-handed helical chirality. The nanotubes are characterized by an extremely high aspect ratio of >1,000 and have a uniform diameter of 20 nm and a wall thickness of 3 nm. The nanotubes with right- and left-handed helical senses were obtained from the (S)- and (R)-enantiomers of the amphiphile, respectively, due to an efficient translation of point chirality into supramolecular helical chirality. The (S)- and (R)-enantiomers coassemble at varying mole ratios to give nanotubes, whose circular dichroism profiles are almost unchanged over a wide range of the enantiomeric excess of the amphiphile (100-20%). The high level of chirality amplification thus observed indicates a long-range cooperativity in the self-assembling process. In sharp contrast, a hexabenzocoronene amphiphile with chiral lipophilic side chains did not form nanotubular assemblies. The present work demonstrates the majority rule in noncovalent systems and also may provide a synthetic strategy toward realization of molecular solenoids. PMID- 16043722 TI - Transfer of glyburide and glipizide into breast milk. AB - OBJECTIVE: To determine if glyburide and glipizide are excreted into breast milk and if breast-feeding from women taking these drugs causes infant hypoglycemia. RESEARCH DESIGN AND METHODS: We studied eight women who had received a single oral dose of 5 or 10 mg glyburide. Drug concentrations were measured in maternal blood and milk for 8 h after the dose. In a separate study, five women were given a daily dosage (5 mg/day) of glyburide or glipizide, starting on the first postpartum day. Maternal blood and milk drug concentrations and infant blood glucose were measured 5-16 days after delivery. RESULTS: In the single-dose glyburide study, the mean maximum theoretical infant dose (MTID) as a percent of the weight-adjusted maternal dose (WAMD) was <1.5 and <0.7% for the 5- and 10-mg doses, respectively. For the five women taking daily dosages, the mean MTID as a percent of the WAMD was <28% for glyburide and <27% for glipizide. The high estimates were due to the insensitivity of the assay. Neither glyburide nor glipizide were detected in breast milk in either study and blood glucose was normal in the three infants (one glyburide and two glipizide) who were wholly breast-fed when the drug concentrations were at steady state. CONCLUSIONS: Neither glyburide nor glipizide were detected in breast milk, and hypoglycemia was not observed in the three nursing infants. Both agents, at the doses tested, appear to be compatible with breast-feeding. PMID- 16043723 TI - Subcutaneous use of a fast-acting insulin analog: an alternative treatment for pediatric patients with diabetic ketoacidosis. AB - OBJECTIVE: To look for technical simplification and economic efficiency in the treatment of pediatric diabetic ketoacidosis (DKA) with subcutaneous use of the fast-acting insulin analog (lispro) and compare its use with regular intravenous insulin treatment. RESEARCH DESIGN AND METHODS: In this controlled clinical trial from June 2001 to June 2003, we randomized 60 episodes of DKA with a blood glucose level > or = 16.6 mmol/l (300 mg/dl), venous pH <7.3 and/or bicarbonate <15 mmol/l, or ketonuria greater than + +. Of the 60 episodes, 30 were treated with subcutaneous lispro (0.15 units/kg) given every 2 h (lispro group) and the other 30 cases received continuous intravenous regular insulin (0.1 unit x kg(-1) x h(-1); CIRI group). Volume deficit was repaired with 10-ml/kg aliquots of 0.9% sodium chloride. Laboratory monitoring included hourly bedside capillary glucose, venous blood gas, beta-hydroxybutyrate, and electrolytes. Plasma blood glucose levels were measured on admission, 2 h after admission, when capillary blood glucose reached < or = 13.8 mmol/l (250 mg/dl), and 6, 12, and 24 h thereafter. RESULTS: Capillary glucose levels decreased by 2.9 and 2.6 mmol x l(-1) x h(-1) in the lispro and CIRI groups, respectively, but blood glucose fluctuated at different time intervals. In the CIRI group, metabolic acidosis and ketosis resolved in the first 6-h period after capillary glucose reached 13.8 mmol/l, whereas in the lispro group, they resolved in the next 6-h interval; however, both groups met DKA recovery criteria without complications. CONCLUSIONS: DKA treatment with a subcutaneous fast-acting insulin analog represents a cost effective and technically simplified procedure that precludes intensive care unit admission. PMID- 16043724 TI - Effects of losartan and amlodipine on urinary albumin excretion and ambulatory blood pressure in hypertensive type 2 diabetic patients with overt nephropathy. AB - OBJECTIVE: Few studies have assessed whether 24-h blood pressure control induced by antihypertensive agents improves macroalbuminuria in hypertensive type 2 diabetic patients with overt nephropathy. We evaluated the effects of losartan and amlodipine on 24-h blood pressure, autonomic nervous activity, and albuminuria in these patients. RESEARCH DESIGN AND METHODS: In this open-label, parallel-prospective, randomized study, 44 patients were treated with losartan and 43 with amlodipine for a 12-week titration phase and a maintenance phase for a maximum of 12 weeks. Twenty-four-hour blood pressure and urinary albumin excretion were measured before and during treatment. Simultaneously, power spectral analysis of heart rate was performed to evaluate low frequency (LF) and high frequency (HF) components and LF-to-HF ratios as an index of sympathovagal balance. RESULTS: Losartan decreased (P < 0.001) mean blood pressure from 162/91 to 150/82 mmHg during daytime and from 146/82 to 137/74 mmHg during nighttime (systolic/diastolic). Amlodipine also decreased (P < 0.001) blood pressure from 159/90 to 147/82 mmHg during daytime and from 143/81 to 131/72 mmHg during nighttime. LF and HF components and nighttime-to-daytime ratios for the LF-to-HF ratios did not differ during treatment in two groups, showing no changes in the diurnal autonomic nervous rhythm. Losartan decreased (P < 0.001) 24-h urinary albumin excretion from 810 mg/day (95% CI 780-1,140) to 570 (510-910). Amlodipine, however, did not decrease (P = 0.893) albuminuria (790 mg/day [780 1,170] vs.790 [710-1,260]). CONCLUSIONS: These results suggest that in type 2 diabetes with overt nephropathy, 24-h blood pressure regulation alone is inadequate to reduce macroalbuminuria and additional effects of losartan are crucial for antiproteinuric action. PMID- 16043725 TI - Foot ulcer risk is lower in South-Asian and african-Caribbean compared with European diabetic patients in the U.K.: the North-West diabetes foot care study. AB - OBJECTIVE: To determine 1) foot ulcer rates for European, South-Asian, and African-Caribbean diabetic patients in the U.K and 2) the contribution of neuropathy and peripheral arterial disease (PAD) differences to altered ulcer risk between the groups. RESEARCH DESIGN AND METHODS: In this U.K. population based study, we screened 15,692 type 1 and type 2 diabetic patients in the community health care setting for foot ulcers, foot deformities, neuropathy, and PAD plus other characteristics. In total, 13,409 were European (85.5%), 1,866 were South Asian (11.9%), and 371 were African Caribbean (2.4%). RESULTS: The age adjusted prevalence of diabetic foot ulcers (past or present) for Europeans, South Asians, and African Caribbeans was 5.5, 1.8, and 2.7%, respectively (P < 0.0001). Asians and African Caribbeans had less neuropathy, PAD, and foot deformities than Europeans (P = 0.003). The unadjusted risk of ulcer (odds ratio [OR]) for Asians versus Europeans was 0.29 (95% CI 0.20-0.41) (P < 0.0001). PAD, neuropathy, foot deformities, and insulin use attenuated the age-adjusted OR from 0.32 to 0.52 (0.35-0.76) (P < 0.0001). African-Caribbean versus European ulcer risk in males was attenuated from 0.60 to 0.71 by vibration sensation. CONCLUSIONS: South Asians with diabetes in the U.K. have about one-third the risk of foot ulcers of Europeans. The lower levels of PAD, neuropathy, insulin usage, and foot deformities of the Asians account for approximately half of this reduced foot ulcer risk. Lower neuropathy is the main contributor to the reduced African Caribbean ulcer rate, particularly in men. The reasons for these ethnic differences warrant further investigation. PMID- 16043726 TI - Annual incidence and clinical characteristics of type 2 diabetes in children as detected by urine glucose screening in the Tokyo metropolitan area. AB - OBJECTIVE: This study investigates the annual incidence and clinical characteristics of type 2 diabetes among school-aged children as detected by urine glucose screening from 1974 to 2002 in the Tokyo metropolitan area. RESEARCH DESIGN AND METHODS: In total, 8,812,356 school children were examined for glucosuria. Morning urine was used for the analysis. When the urine was positive for glucose, an oral glucose tolerance test was carried out to confirm diabetes. RESULTS: In all, 232 students were identified to have type 2 diabetes. The overall annual incidence of type 2 diabetes was 2.63/100,000. The annual incidence after 1981 was significantly higher than that before 1980 (1.73 vs. 2.76/100,000, P < 0.0001). The annual incidence was significantly higher for junior high school students compared with primary school students (0.78 vs. 6.43/100,000, P < 0.0001). The overall male-to-female ratio of students with type 2 diabetes was 1.0:1.19 (P = 0.296), but it was 1.0:1.56 (P = 0.278) for primary school students. Overall, 83.4% of children with diabetes were obese (> or = 20% overweight). However, nonobese girls (<20% overweight) with diabetes accounted for 23.0% of the patients, whereas markedly obese boys (> or = 40% overweight) accounted for 61.5% of the patients. The frequency of a family history of type 2 diabetes in second- and first-degree relatives was 56.5%. CONCLUSIONS: We confirmed that the incidence of young people with type 2 diabetes increased after 1981 in the Tokyo metropolitan area. The increase in the frequency of this disorder seemed to be strongly related to an increasing prevalence of obesity. Age and genetic susceptibility may be associated with the occurrence of type 2 diabetes. PMID- 16043727 TI - Family history of type 2 diabetes is associated with increased carotid artery intimal-medial thickness in Mexican Americans. AB - OBJECTIVE: To evaluate whether the joint risk of diabetes and atherosclerosis tracked within families, we assessed the correlation between a family history of diabetes and intimal-medial thickness (IMT) of the common carotid artery (CCA). RESEARCH DESIGN AND METHODS: Study subjects included 620 nondiabetic individuals from 24 families enrolled in the San Antonio Family Heart Study. The thickness of the far walls of the CCA was measured by B-mode ultrasonography. Statistical analyses included familial correlations to account for the nonindependence of family data. RESULTS: After adjusting for sex, age, and IMT reader effects, the heritability of IMT in this population was 16% (P = 0.009). Using a more comprehensive family history score that accounted for diabetes status of the individual's parents and older siblings, we observed a significant correlation between family history score and increased CCA IMT (0.006 mm increase in CCA IMT for every point increase of diabetes family history score, P = 0.016). This association remained even after further adjustment for BMI, smoking, and fasting insulin and glucose levels. After adjusting for several cardiovascular risk factors, the mean CCA IMT in those with high family history scores for diabetes was still 0.037 mm thicker than those with low family history scores for diabetes (P = 0.040). CONCLUSIONS: These results suggest that the genetic contribution to CCA IMT variability is modest. Also, the small increase in subclinical atherosclerosis observed even among nondiabetic Mexican Americans with a positive family history of diabetes is probably transmitted along with the risk of diabetes through shared etiologic risk factors between diabetes and cardiovascular disease. PMID- 16043728 TI - Randomized trial of quality improvement intervention to improve diabetes care in primary care settings. AB - OBJECTIVE: To assess the impact of a quality improvement (QI) intervention on the quality of diabetes care at primary care clinics. RESEARCH DESIGN AND METHODS: Twelve primary care medical practices were matched by size and location and randomized to intervention or control conditions. Intervention clinic staff were trained in a seven-step QI change process to improve diabetes care. Surveys and medical record reviews of 754 patients, surveys of 329 clinic staff, interviews with clinic leaders, and analysis of training session videotapes evaluated compliance with and impact of the intervention. Mixed-model nested analyses compared differences in the quality of diabetes care before and after intervention. RESULTS: All intervention clinics completed at least six steps of the seven-step QI change process in an 18-month period and, compared with control clinics, had broader staff participation in QI activities (P = 0.04), used patient registries more often (P = 0.03), and had better test rates for HbA(1c) (A1C), LDL, and blood pressure (P = 0.02). Other processes of diabetes care were unchanged. The intervention did not improve A1C (P = 0.54), LDL (P = 0.46), or blood pressure (P = 0.69) levels or a composite of these outcomes (P = 0.35). CONCLUSIONS: This QI change process was successfully implemented but failed to improve A1C, LDL, or blood pressure levels. Data suggest that to be successful, such a QI change process should direct more attention to specific clinical actions, such as drug intensification and patient activation. PMID- 16043729 TI - Depressive symptoms in mothers of infants identified as genetically at risk for type 1 diabetes. AB - OBJECTIVE: This study describes maternal depression associated with newborn genetic screening for type 1 diabetes after risk notification. RESEARCH DESIGN AND METHODS: Mothers of at-risk infants (n = 192), identified through newborn genetic screening as part of the Prospective Assessment of Newborns for Diabetes Autoimmunity study, were administered a structured telephone interview assessing maternal depressive symptoms 1 and 3.5 months after risk notification. Statistical analyses were conducted to examine predictors and correlates of maternal depressive symptoms. RESULTS: For the total sample, maternal depressive symptoms in response to infant risk status were not elevated at 1 and 3.5 months after risk notification. However, at the first interview, mothers from ethnic minority backgrounds (P < 0.002), with limited education (P < 0.001), and with postpartum depression symptomatology (P < 0.001) reported significantly more depressive symptoms in response to risk notification (r2 = 0.354). At the second interview, postpartum depression symptomatology remained a powerful predictor of depressive symptoms in response to risk notification (P < 0.001). In addition, certain coping styles (wishful thinking, self-blame, and seeking social support) were associated with increased depressive symptoms. A history of major depression was a correlate of both postpartum depressive symptomatology (r = 0.26) and maternal depressive response to risk notification (r = 0.21). CONCLUSIONS: For the most part, mothers of infants genetically at risk for type 1 diabetes do not appear to evidence elevated depressive symptoms. This suggests that most mothers are resilient when notified of infant risk. However, certain maternal characteristics such as ethnic minority status, less than a high school education, postpartum depression symptomatology, a history of major depression, and certain coping strategies (wishful thinking, self-blame, and seeking social support) appear to be associated with a more difficult maternal response to the news of an infant's increased genetic risk for type 1 diabetes. PMID- 16043730 TI - Depression and diabetes: a large population-based study of sociodemographic, lifestyle, and clinical factors associated with depression in type 1 and type 2 diabetes. AB - OBJECTIVE: The purpose of this study was to investigate factors associated with depression in type 1 and type 2 diabetes and test whether these differ from factors associated with depression in the nondiabetic population. RESEARCH DESIGN AND METHODS: In an unselected population study comprising 60,869 individuals, potential sociodemographic, lifestyle, and clinical factors were investigated in participants with and without diabetes. The associations between hyperglycemia and depression in types 1 and 2 diabetes were also studied. The levels of depression were self-rated by using the Hospital Anxiety and Depression Scale. RESULTS: Several factors were correlated with depression in types 1 and 2 diabetes. However, these factors were not different from those of the nondiabetic population. Comorbid chronic somatic diseases were associated with depression in type 2 but not type 1 diabetes. In type 2 diabetes, those without comorbidity had the same odds of depression as the nondiabetic population with no chronic somatic diseases. No significant associations were found for hyperglycemia in relation to depression in type 1 and type 2 diabetes. CONCLUSIONS: Type 2 diabetes without other chronic somatic diseases did not increase the risk of depression. Factors associated with depression in type 1 and type 2 diabetes were shared with the nondiabetic population. PMID- 16043731 TI - Acute psychological stress affects glucose concentrations in patients with type 1 diabetes following food intake but not in the fasting state. AB - OBJECTIVE: To compare the effect of acute psychosocial stress on glucose concentrations in the fasting state and following food intake in patients with type 1 diabetes. RESEARCH DESIGN AND METHODS: In study 1, 20 patients were exposed to moderate psychosocial stress by means of the Trier Social Stress Test (TSST) in the fasting state. In study 2, the TSST was applied to 20 additional patients 75 min after intake of a standard meal. Glucose concentrations (by continuous glucose monitoring system), blood pressure, and heart rate were monitored on the control day and on the stress testing day. RESULTS: In both studies, blood pressure increased in response to TSST from 122/77 +/- 14/9 mmHg at baseline to a maximum of 152/93 +/- 21/13 mmHg (P < 0.001), and heart rate increased from 80 +/- 11 to 99 +/- 19 bpm (P < 0.001). In the fasting state (study 1), glucose concentrations remained unchanged during the control day as well as during the stress testing day. In study 2, glucose concentrations were similar on both days before and up to 75 min after the intake of the standard meal. However, a significant delay (of 45 min) in the decrease of glucose concentrations was induced by psychological stress. A two-factor repeated measures ANOVA revealed a significant difference of glucose concentrations over time (F = 646.65/P < 0.001). CONCLUSIONS: In the postprandial period, acute psychological stress induced a significantly delayed decrease of glucose concentrations, whereas in the fasting state, no effect on poststress glucose concentrations was observed. PMID- 16043732 TI - Joint distribution of non-HDL and LDL cholesterol and coronary heart disease risk prediction among individuals with and without diabetes. AB - OBJECTIVE: To assess coronary heart disease (CHD) risk within levels of the joint distribution of non-HDL and LDL cholesterol among individuals with and without diabetes. RESEARCH DESIGN AND METHODS: We used four publicly available data sets for this pooled post hoc analysis and confined the eligible subjects to white individuals aged > or = 30 years and free of CHD at baseline (12,660 men and 6,721 women). Diabetes status was defined as either "reported by physician diagnosed and on medication" or having a fasting glucose level > or = 126 mg/dl at the baseline examination. The primary end point was CHD death. Within diabetes categories, risk was assessed based on lipid levels (in mg/dl): non-HDL <130 and LDL <100 (group 1); non-HDL <130 and LDL > or = 100 (group 2); non-HDL > or = 130 and LDL <100 (group 3); and non-HDL > or = 130 and LDL > or = 100 (group 4). Group 1 within those without diabetes was the overall reference group. RESULTS: Of the subjects studied, approximately 6% of men and 4% of women were defined as having diabetes. A total of 773 CHD deaths occurred during the average 13 years of follow-up time. A Cox proportional hazard model was used to estimate the relative risk (RR) of CHD death. Those with diabetes had a 200% higher RR than those without diabetes. In a multivariate model, CHD risk in those with diabetes did not increase with increasing LDL, whereas it did increase with increasing non HDL: RR (95% confidence interval) for group 1: 5.7 (2.0-16.8); group 2: 5.7 (1.6 20.7); group 3: 7.2 (2.6-19.8); and group 4: 7.1 (3.7-13.6). CONCLUSIONS: Non-HDL is a stronger predictor of CHD death among those with diabetes than LDL and should be given more consideration in the clinical approach to risk reduction among diabetic patients. PMID- 16043733 TI - Efficacy of inhaled insulin in patients with type 2 diabetes not controlled with diet and exercise: a 12-week, randomized, comparative trial. AB - OBJECTIVE: Effective type 2 diabetes management requires prompt intervention if glycemic control is not achieved by nonpharmacological means. This study investigates whether inhaled insulin (INH; Exubera) can achieve target glycemic control in patients failing on diet and exercise. RESEARCH DESIGN AND METHODS: Patients with suboptimal control on diet and exercise (HbA(1c) [A1C] 8-11%) were randomized to 3 months' treatment with either INH before meals (n = 76) or rosiglitazone 4 mg twice a day (n = 69), in conjunction with a diet and exercise regimen. The primary end point was percentage of patients achieving A1C <8.0%. RESULTS: The INH and rosiglitazone groups had comparable baseline A1C values (9.5 vs. 9.4%, respectively). Significantly more patients achieved A1C <8.0% (83 vs. 58%, adjusted odds ratio 7.14 [95% CI 2.48-20.58], P = 0.0003), A1C <7.0% (44 vs. 18%, 4.43 [1.94-10.12]), and A1C < or = 6.5% (28 vs. 7.5% 5.34 [1.83-15.57]) with INH. A1C decrease was greater with INH (-2.3% vs. -1.4%, adjusted treatment group difference: -0.89% [95% CI -1.23 to -0.55]) with final mean A1C values of 7.2 and 8.0% for INH and rosiglitazone, respectively. Hypoglycemia (episodes per subject month) was higher with INH (0.7 vs. 0.05, risk ratio 14.72 [95% CI 7.51-28.83]), with no severe hypoglycemic episodes. Pulmonary function changes were small and comparable between groups. CONCLUSIONS: INH could be an effective therapy for people with type 2 diabetes early in the course of their disease. PMID- 16043734 TI - Youth and parent satisfaction with clinical use of the GlucoWatch G2 Biographer in the management of pediatric type 1 diabetes. AB - OBJECTIVE: A continuous glucose monitor satisfaction scale (CGM-SAT) was evaluated during a 6-month randomized controlled trial of the GlucoWatch G2 Biographer (GW2B) in youths with type 1 diabetes. RESEARCH DESIGN AND METHODS: At the end of the 6-month trial, 97 parents and 66 older children who had been randomized to the GW2B group completed the CGM-SAT, which assesses satisfaction on 37 items using a five-point Likert scale. Descriptive analysis, calculation of several reliability estimates, and assessment of concurrent validity were performed. RESULTS: The CGM-SAT demonstrated high internal reliability (Cronbach's alpha = 0.95 for parents and 0.94 for youths aged > or = 11 years), split-half reliability (rho = 0.91 for parents and 0.93 for youths), and parent adolescent agreement (rho = 0.68, P < 0.001). Convergent validity was supported by marginally significant associations with treatment adherence and frequency of GW2B use. CGM-SAT scores did not correlate significantly with changes in treatment adherence, quality of life, or diabetes-related anxiety from baseline to 6 months. Mean scores on CGM-SAT items indicated that 81% of parental responses and 73% of youths' responses were less favorable than "neutral." Descriptive analysis indicated the GW2B requires substantial improvement before it can achieve widespread clinical utility and acceptance. CONCLUSIONS: The results supported the psychometric properties of the CGM-SAT. The CGM-SAT warrants further research use and cross-validation with other continuous glucose monitors. This study provides a benchmark for comparison with new glucose sensors. PMID- 16043735 TI - Improved meal-related beta-cell function and insulin sensitivity by the dipeptidyl peptidase-IV inhibitor vildagliptin in metformin-treated patients with type 2 diabetes over 1 year. AB - OBJECTIVE: To examine the effects of dipeptidyl peptidase-IV (DPP-4) inhibition on meal-related beta-cell function and insulin sensitivity over 52 weeks in type 2 diabetes. RESEARCH DESIGN AND METHODS: In a 12-week core study, placebo (n = 51) or vildagliptin (n = 56; 50 mg OD) was added to metformin treatment (1.5-3.0 mg/day). A 40-week extension followed in 71 patients. Meal tests were performed at 0, 12, 24, and 52 weeks; glucose, insulin, and C-peptide were evaluated. RESULTS: In subjects completing 52 weeks with participation in all meal tests (n = 57), HbA(1c) (A1C) decreased in the vildagliptin/metformin group (VM group, n = 31) but increased in the placebo/metformin group (PM group, n = 26; between-group difference -1.0 +/- 0.2%; P < 0.001; baseline of all subjects combined 7.7 +/- 0.1%). Also, fasting glucose decreased in the VM group but increased in the PM group (difference -0.9 +/- 0.3 mmol/l, P = 0.016; baseline 9.8 +/- 0.3 mmol/l). Insulin secretion (postmeal suprabasal area under the 0- to 30-min C-peptide curve divided by the 30-min increase in glucose) was increased in the VM group but was reduced in the PM group (difference +0.011 +/- 0.03 pmol/l 30 min/mmol/l, P = 0.018; baseline 0.036 +/- 0.02). Insulin sensitivity during meal ingestion (oral glucose insulin sensitivity) increased in the VM group but was not altered in the PM group (difference +27 +/- 4 ml x min(-1) x m(-2), P = 0.036; baseline 246 +/- 6). Insulin secretion related to insulin sensitivity (adaptation index) increased in the VM group but decreased in the PM group (difference +3.2 +/- 1.0, P = 0.040; baseline 9.1 +/- 0.5). The change in adaptation index correlated to the change in A1C (r = -0.39, P = 0.004). CONCLUSIONS: This study presents evidence that DPP-4 inhibition by vildagliptin when added to metformin in type 2 diabetes over 52 weeks improves beta-cell function along with improved postmeal insulin sensitivity. PMID- 16043736 TI - Somatostatin molecular variants in the vitreous fluid: a comparative study between diabetic patients with proliferative diabetic retinopathy and nondiabetic control subjects. AB - OBJECTIVE: There is growing evidence to indicate that somatostatin could be added to the list of natural antiangiogenic factors that exist in the vitreous fluid. In addition, a deficit of intravitreous somatostatin-like immunoreactivity (SLI) has been found in diabetic patients with proliferative diabetic retinopathy (PDR). In the present study, we have determined the main molecular variants of somatostatin (somatostatin-14 and somatostatin-28) in the vitreous fluid and plasma of nondiabetic control subjects and diabetic patients with PDR. In addition, the contribution of cortistatin, a neuropeptide with strong structural similarities to somatostatin, to SLI and its levels in vitreous and plasma in both nondiabetic and diabetic patients has also been measured. RESERCH DESIGN AND METHODS: Plasma and vitreous fluid from 22 diabetic patients with PDR and 22 nondiabetic control subjects were analyzed. Somatostatin-14, somatostatin-28 and cortistatin were measured by radioimmunoassay but separation by high-performance liquid chromatography was required to measure somatostatin-14. RESULTS: The predominant molecular form of somatostatin within the vitreous fluid was somatostatin-28 (fivefold higher than somatostatin-14 in control subjects and threefold higher in patients with PDR). Cortistatin significantly contributed to SLI and its intravitreous levels were higher than those detected in plasma (nondiabetic control subjects: 147 [102-837] vs. 78 [24-32] pg/ml; patients with PDR: 187 [87-998] vs. 62 [24-472] pg/ml; P = 0.01 for both). Intravitreous somatostatin-14 was similar in both subjects with PDR and the control group (P = 0.87). By contrast, somatostatin-28 concentration was lower in patients with PDR than in nondiabetic control subjects (350 +/- 32 vs. 595 +/- 66 pg/ml; P = 0.004). CONCLUSIONS: Somatostatin-28 is the main molecular variant in the vitreous fluid. The intravitreous SLI deficit detected in patients with PDR is mainly due to somatostatin-28. Cortistatin is abundant in the vitreous fluid and significantly contributes to SLI. These findings could open up new strategies for PDR treatment. PMID- 16043737 TI - Effect of low-dose insulin treatment on body weight and physical development in children and adolescents at risk for type 1 diabetes. AB - OBJECTIVE: Insulin's role in body weight regulation is controversial. We evaluated the effect of parenteral insulin on body weight and physical development in children and adolescents at risk for type 1 diabetes. RESEARCH DESIGN AND METHODS: We performed a secondary analysis of the parenteral arm of the Diabetes Prevention Trial-Type 1 Diabetes (DPT-1), a randomized controlled trial of low-dose parenteral insulin (human ultralente insulin at 0.25 units x kg(-1) x day(-1)) in subjects with a >50% 5-year risk of diabetes. Analysis was limited to 100 subjects (55 intervention, 45 closely monitored) aged <19 years at randomization whose weight was followed for at least 2 years by study end after excluding subjects who were noncompliant within 2 years or developed diabetes within 36 months of randomization. RESULTS: Subjects ranged in age from 4.07 to 18.98 years. There were no significant differences at randomization between subjects in each group with respect to sex, age, weight, height, BMI, Tanner stage, or glucose tolerance. We found no differences over 2 years between the intervention and closely monitored groups in the change in weight (median 6.8 vs. 6.0 kg, P = 0.65), height (median 10.7 vs. 10.1 cm, P = 0.66), BMI (median 0.9 vs. 1.0 kg/m2, P = 0.79), or Tanner stage (median 0 vs. 0, P = 0.35). Multiple regression showed no effect of insulin on change in weight (P = 0.53) or BMI (P = 0.95) over 2 years after adjustment for relevant covariates. CONCLUSIONS: Low dose insulin treatment for 2 years did not affect the weight, BMI, or physical development of nondiabetic children and adolescents. PMID- 16043738 TI - Ethnicity, race, and baseline retinopathy correlates in the veterans affairs diabetes trial. AB - OBJECTIVE: The Veterans Affairs Diabetes Trial (VADT) cohort is enriched with approximately 20% Hispanics and 20% African Americans, affording a unique opportunity to study ethnic differences in retinopathy. RESEARCH DESIGN AND METHODS: Cross-sectional analyses on the baseline seven-field stereo fundus photos of 1,283 patients are reported here. Diabetic retinopathy scores are grouped into four classes of increasing severity: none (10-14), minimal nonproliferative diabetic retinopathy (NPDR) (15-39), moderate to severe NPDR (40 59), and proliferative diabetic retinopathy (60+). These four groups have also been dichotomized to none or minimal (10-39) and moderate to severe diabetic retinopathy (40+). RESULTS: The prevalence of diabetic retinopathy scores >40 was higher for Hispanics (36%) and African Americans (29%) than for non-Hispanic whites (22%). The difference between Hispanics and non-Hispanic whites was significant (P < 0.05). Similarly, the prevalence of diabetic retinopathy scores >40 was significantly higher in African Americans than in non-Hispanic whites (P < 0.05). These differences could not be accounted for by an imbalance in traditional risk factors such as age, duration of diagnosed diabetes, HbA(1c) (A1C), and blood pressure. Diabetic retinopathy severity scores were also significantly associated with increasing years of disease duration, A1C, systolic and diastolic blood pressure, the degree of microalbuminuria, fibrinogen, and the percentage of patients with amputations. There was no relationship between retinopathy severity and the percentage of people who had strokes or cardiac revascularization procedures. There was an inverse relationship between retinopathy severity and total cholesterol, triglycerides, and plasminogen activator inhibitor-1 as well as with smoking history. Diabetic retinopathy scores were not associated with age. CONCLUSIONS: In addition to many well-known associations with retinopathy, a higher frequency of severe diabetic retinopathy was found in the Hispanic and African-American patients at entry into the VADT that is not accounted for by traditional risk factors for diabetic retinopathy, and these substantial ethnic differences remain to be explained. PMID- 16043739 TI - Autoantibodies to autonomic nerves associated with cardiac and peripheral autonomic neuropathy. AB - OBJECTIVE: This study examines whether autonomic nerve autoantibodies (ANabs) are associated with development of autonomic neuropathy using a prospective study design. RESEARCH DESIGN AND METHODS: A group of type 1 diabetic patients were followed prospectively with regard to autonomic nerve function on four occasions. At the third examination, 41 patients were tested for ANabs (complement-fixing autoantibodies to the sympathetic ganglion, vagus nerve, and adrenal medulla), and the results were related to cardiac autonomic nerve function (heart rate variation during deep breathing [expiration/inspiration ratio] and heart-rate reaction to tilt [acceleration and brake index]) and to peripheral sympathetic nerve function (vasoconstriction after indirect cooling [vasoconstriction index]). RESULTS: ANabs were detected in 23 of 41 (56%) patients at the third examination. Compared with patients without ANabs (ANabs-), patients with ANabs (ANabs+) showed significantly higher frequencies of at least one abnormal cardiac autonomic nerve function test at the third examination (17 of 23 [74%] vs. 7 of 18 [39%]; P = 0.03) and fourth examination (15 of 21 [71%] vs. 4 of 16 [25%]; P < 0.01). In contrast, there was no similar difference at the first or second examination. The relative risk for ANabs(+) patients to develop cardiac autonomic neuropathy at follow-up was 7.5 (95% CI 1.72-32.80). The vasoconstriction index was more abnormal in ANabs+ than in ANabs- patients at the fourth examination (median 1.40 [interquartile range 1.58] vs. 0.35 [2.05]; P = 0.01). CONCLUSIONS: ANabs were associated with future development of cardiac and peripheral autonomic neuropathy in diabetic patients, implying an etiological relationship between nervous tissue autoimmunity and these diabetes complications. PMID- 16043740 TI - Glycemic control, atherosclerosis, and risk factors for cardiovascular disease in individuals with diabetes: the atherosclerosis risk in communities study. AB - OBJECTIVE: Glycemic control (HbA(1c) [A1C]) is strongly associated with microvascular disease in individuals with diabetes, but its relation to macrovascular disease and atherosclerosis is less clear. This study examines the relationship between A1C, carotid intima-media thickness (IMT), and traditional cardiovascular risk factors in individuals with diabetes. RESEARCH DESIGN AND METHODS: A cross-sectional study of 2,060 people with diagnosed and undiagnosed (unrecognized) diabetes in the Atherosclerosis Risk in Communities study was performed. RESULTS: LDL and HDL cholesterol, plasma triglycerides, and waist-to hip ratio were significantly associated with A1C after multivariable adjustment. African Americans with undiagnosed and diagnosed diabetes had significantly elevated A1C values compared with whites, even after adjustment for potentially confounding factors. There was a graded association between A1C and carotid IMT. In a fully adjusted model in individuals with undiagnosed diabetes, the odds ratio (OR) of being in the highest quartile of IMT versus the lowest was 2.46 (95% CI 1.16-5.03, comparing the highest quartile of A1C to the lowest). In people with diagnosed diabetes, the comparable OR was 2.62 (1.36-5.06). CONCLUSIONS: This study identified several important associations between A1C and known risk factors for cardiovascular disease and suggested that A1C is independently related to carotid IMT. Chronically elevated glucose levels may contribute to the development of atherosclerosis in people with diabetes, independent of other risk factors. PMID- 16043741 TI - Discordant trends in microvascular complications in adolescents with type 1 diabetes from 1990 to 2002. AB - OBJECTIVE: Since the Diabetes Control and Complications Trial, diabetes management goals have changed. The aims of the present study were to assess complication rates, including nerve abnormalities, in adolescents from 1990 to 2002 and to investigate associated risk factors. RESEARCH DESIGN AND METHODS: Cross-sectional analysis of complications was assessed in three study periods (1990-1994 [T1], 1995-1998 [T2], and 1999-2002 [T3]) in adolescents matched for age and diabetes duration (n = 878, median age 14.6 years, median duration 7.5 years). Retinopathy was assessed by seven-field stereoscopic fundal photography, albumin excretion rate (AER) from three consecutive timed overnight urine collections, peripheral nerve function by thermal and vibration thresholds, and autonomic nerve function by cardiovascular reflexes. RESULTS: Retinopathy declined significantly (T1, 49%; T2, 31%; and T3, 24%; P < 0.0001), early elevation of AER (> or = 7.5 microg/min) declined (38, 30, and 25%, respectively, P = 0.022), and microalbuminuria (AER > or = 20 microg/min) declined (7, 3, and 3%, respectively; P = 0.017, T1 vs. T2 and T3). Autonomic nerve abnormalities were unchanged (18, 21, and 18%, respectively; P = 0.60), but peripheral nerve abnormalities increased (12, 19, and 24%, respectively; P = 0.0017). More patients were treated with three or more injections per day (12, 46, and 67%, respectively; P < 0.0001) and insulin dose increased (1.08, 1.17, and 1.22 units x kg(-1) x day(-1), respectively; P < 0.0001), but median HbA(1c) (A1C) was unchanged (8.5, 8.5, and 8.4%, respectively). BMI and height SD score increased: BMI 0.46, 0.67, and 0.79, respectively (P < 0.0001), and height -0.09, 0.05, and 0.27, respectively (P < 0.0001). CONCLUSIONS: Retinopathy and microalbuminuria declined over time in this cohort, but the increased rate of peripheral nerve abnormalities is of concern. Despite intensified management (higher insulin dose and more injections), A1C has not changed and remains well above the recommended targets for adolescents. PMID- 16043742 TI - Relationship between HbA1c level and peripheral arterial disease. AB - OBJECTIVE: Homeostatic glucose control may play an important role in the development of peripheral arterial disease among individuals without diabetes. We sought to evaluate the association of HbA(1c) (A1C) with peripheral arterial disease in a representative sample of the U.S. population with and without diabetes. RESEARCH DESIGN AND METHODS: A cross-sectional study was conducted among 4,526 National Health and Nutrition Examination Survey 1999-2002 participants > or = 40 years of age. Peripheral arterial disease was defined as an ankle-brachial index <0.9 (n = 327). RESULTS: Among nondiabetic subjects, the age-standardized prevalence of peripheral arterial disease was 3.1, 4.8, 4.7, and 6.4% for participants with an A1C <5.3, 5.3-5.4, 5.5-5.6, and 5.7-6.0%, respectively (P trend <0.001). The prevalence of peripheral arterial disease was 7.5 and 8.8% for diabetic participants with A1C <7 and > or = 7%, respectively. After multivariable adjustment and compared with nondiabetic participants with A1C <5.3%, the odds ratio (95% CI) of peripheral arterial disease for nondiabetic participants with an A1C of 5.3-5.4, 5.5-5.6, and 5.7-6.0% was 1.41 (0.85-2.32), 1.39 (0.70-2.75), and 1.57 (1.02-2.47), respectively, and it was 2.33 (1.15-4.70) and 2.74 (1.25-6.02) for diabetic participants with A1C <7 and > or = 7%, respectively. CONCLUSIONS: An association exists between higher levels of A1C and peripheral arterial disease, even among patients without diabetes. Individuals with A1C levels > or = 5.3% should be targeted for aggressive risk factor reduction, which may reduce the burden of subclinical cardiovascular disease even among those without diabetes. PMID- 16043743 TI - Oral contraceptives, angiotensin-dependent renal vasoconstriction, and risk of diabetic nephropathy. AB - OBJECTIVE: Diabetes, the leading cause of end-stage renal disease in the U.S., is believed to involve activation of the renin angiotensin system (RAS) as a risk factor for nephropathy. RAS activation occurs in healthy women using oral contraceptives (OCs), but the effects of OC use on the diabetic kidney are unclear. RESEARCH DESIGN AND METHODS: Renal plasma flow (RPF) response to captopril, as an index of RAS activity, was investigated in 92 women (41 nondiabetic OC nonusers, 10 nondiabetic OC users, 29 diabetic OC nonusers, and 12 diabetic OC users). Based on the hemodynamic findings, we examined the impact of OC use on the development of nephropathy as a post hoc analysis in an inception cohort of 114 female patients with newly diagnosed type 1 diabetes followed for a median of 20.7 years (range 1-24). RESULTS: Nondiabetic OC nonusers showed minimal RPF vasodilator response to captopril (9 +/- 10 ml x min(-1) x 1.73 m( 2), P = 0.6). In comparison, nondiabetic OC users showed a significant increase (69 +/- 35 ml x min(-1) x 1.73 m(-2), P = 0.02) (P = 0.04 vs. nondiabetic OC nonusers). Diabetic OC nonusers demonstrated the anticipated vasodilator response (58 +/- 12 ml x min(-1) x 1.73 m(-2), P < 0.0001). Diabetic OC users showed the largest responses (84 +/- 12 ml x min(-1) x 1.73 m(-2), P = 0.002) (P = 0.04 vs. diabetic OC nonusers). Plasma renin activity did not vary with OC use (P = 0.3). The RPF responses to captopril and angiotensin receptor blocker were highly correlated (r = 0.72, P < 0.001), suggesting clear involvement of the RAS. In the observational study, 18% (6/33 [95% CI 4.3-32.1]) of OC users developed macroalbuminuria compared with 2% (2/81 [0-5.9]) of OC nonusers (P = 0.003, univariate analysis). After adjustment for known risk factors with a Cox regression model, OC use remained a predictor for the development of macroalbuminuria (relative risk 8.90 [95%CI 1.79-44.36], P = 0.008). CONCLUSIONS: The strong association of OC use with angiotensin-dependent control of the renal circulation and the development of macroalbuminuria suggest that OC use may be a risk factor for diabetic nephropathy. Large prospective studies are required to further investigate this relationship. PMID- 16043744 TI - Insulin resistance and preeclampsia in gestational diabetes mellitus. AB - OBJECTIVE: To compare the degree of insulin resistance in women with gestational diabetes mellitus (GDM) who do and do not develop preeclampsia. RESEARCH DESIGN AND METHODS: We conducted a prospective cohort study of initially normotensive women with GDM who underwent oral glucose tolerance tests (OGTTs), intravenous glucose tolerance tests (IVGTTs), and glucose clamp studies in the early third trimester (n = 150) and 15 months postpartum (n = 89). After delivery, the women were categorized as nonpreeclamptic or preeclamptic (systolic blood pressure [SBP] > or = 140 mmHg, diastolic blood pressure [DBP] > or = 90 mmHg, and at least >1+ proteinuria or >300 mg/24 h). Metabolic parameters between the groups were compared by chi2 or Fisher's exact tests and ANOVA with P < 0.05 as significant. RESULTS: A total of 29 women (19%) developed preeclampsia, which was mild in 21 and severe in 8 women. At entry, there were no differences in age, weight indexes, and glycemic measures between the nonpreeclamptic and preeclamptic groups. Those with preeclampsia were significantly taller (61.5 +/- 2.4 vs. 60.1 +/- 2.3 in, P = 0.003), were more often nulliparous (38 vs. 16%, P = 0.01), and had higher entry SBP (112 +/- 10 vs. 103 +/- 6.9 mmHg, P < 0.0001) and DBP (64 +/- 9 vs. 59 +/- 5 mmHg, P = 0.002). No significant differences between the groups were found in any measures of the OGTT glucose levels, insulin sensitivity index, glucose effectiveness, acute response to glucose, or disposition index, nor were there any differences found in the euglycemic clamp measures of basal or steady-state levels of glucose, insulin, free fatty acid, hepatic glucose output, peripheral glucose clearance, C-peptide, or glucagon. At 15 months postpartum, blood pressure levels remained significantly higher in the preeclamptic group (n = 19) compared with the nonpreeclamptic group (n = 70). No differences in any glycemic or insulin resistance measures were found. CONCLUSIONS: Women with GDM were uniformly insulin resistant. Those who developed preeclampsia, when compared with those who remained nonpreeclamptic, were not more insulin resistant in either the third trimester or 15 months postpartum. However, women who developed preeclampsia had blood pressure levels that were significantly higher, although still in the normal range, than those of women who remained nonpreeclamptic. PMID- 16043745 TI - Efficacy of multilayered hosiery in reducing in-shoe plantar foot pressure in high-risk patients with diabetes. AB - OBJECTIVE: High plantar pressure is an acknowledged risk factor in the development of plantar ulcers in the diabetic neuropathic foot. This study examines the ability of preventive foot care (PFC) socks to reduce plantar foot pressures in a sample of high-risk patients with diabetes. RESEARCH DESIGN AND METHODS: Nineteen patients with established peripheral neuropathy attending a complications clinic of the Manchester Diabetes Centre were recruited to the study. Fifteen (78%) of the patients were male, 40-80 years of age, and ulcer free at the time of recruitment. In-shoe plantar pressure measurements were recorded using the F-Scan and compared PFC socks with ordinary supermarket socks. The analysis measured differences in maximum foot contact area and plantar pressure for the whole foot, forefoot, and peak plantar pressure areas. RESULTS: The results showed a significant increase in maximum foot contact area of 11 cm2 (95% CI 7-11) when subjects wore the PFC socks (P < 0.01). This was accompanied by 5.4 kPa (3.5-7.3) or 9% reduction in total foot pressure (P < 0.01). Similar results were observed at the forefoot, which showed a 14.2% increase in contact area and a 10.2% reduction in peak forefoot pressure. CONCLUSIONS: These results suggest that the wearing of PFC socks increases the underfoot contact area and hence decreases plantar foot pressures. Further studies are required to determine whether the pressure and friction reductions achieved by this simple intervention would be effective in reducing the incidence of foot ulcers in high-risk patients. PMID- 16043746 TI - C-174G polymorphism in the promoter of the interleukin-6 gene is associated with insulin resistance. AB - OBJECTIVE: The C-174G promoter polymorphism of the interleukin (IL)-6 gene was found to influence transcriptional activity and plasma IL-6 levels in humans. We addressed the question of whether the C-174G IL-6 polymorphism contributes to variation of insulin sensitivity. RESEARCH DESIGN AND METHODS: Two cohorts of subjects were genotyped. Cohort 1 includes 275 nondiabetic subjects who underwent a euglycemic-hyperinsulinemic clamp. Cohort 2 includes 77 patients with morbid obesity who underwent laparoscopic adjustable gastric banding (LAGB). RESULTS: The genotypes were consistent with Hardy-Weinberg equilibrium proportions. In cohort 1, insulin sensitivity was reduced in carriers of the -174G/G genotype as compared with subjects carrying the C allele (P = 0.004). Carriers of -174G/G displayed significantly higher plasma IL-6 levels in comparison with carriers of the C allele. In a stepwise linear regression analysis, the C-174G polymorphism was independently associated with insulin sensitivity; however, after inclusion of plasma IL-6 concentrations, the polymorphism was excluded from the model explaining insulin sensitivity variability, thus suggesting that the polymorphism was affecting insulin sensitivity by regulating IL-6 plasma levels. IL-6 mRNA levels were measured by real-time RT-PCR in subcutaneous fat obtained from obese patients of cohort 2 during LAGB. Carriers of -174G/G showed increased IL-6 expression compared with subjects carrying the C allele (P = 0.04). There was a significant correlation between adipose IL-6 mRNA expression and insulin resistance assessed by homeostasis model assessment (rho = 0.28, P = 0.014). CONCLUSIONS: These results indicate that the -174G/G genotype of the IL-6 gene may contribute to variations in insulin sensitivity. PMID- 16043747 TI - Identifying individuals at high risk for diabetes: The Atherosclerosis Risk in Communities study. AB - OBJECTIVE: To develop and evaluate clinical rules to predict risk for diabetes in middle-aged adults. RESEARCH DESIGN AND METHODS: The Atherosclerosis Risk in Communities is a cohort study conducted from 1987-1989 to 1996-1998. We studied 7,915 participants 45-64 years of age, free of diabetes at baseline, and ascertained 1,292 incident cases of diabetes by clinical diagnosis or oral glucose tolerance testing. RESULTS: We derived risk functions to predict diabetes using logistic regression in a random half of the sample. Rules based on these risk functions were evaluated in the other half. A risk function based on waist, height, hypertension, blood pressure, family history of diabetes, ethnicity, and age was performed similarly to one based on fasting glucose (area under the receiver-operating characteristic curve [AUC] 0.71 and 0.74, respectively; P = 0.2). Risk functions composed of the clinical variables plus fasting glucose (AUC 0.78) and additionally including triglycerides and HDL cholesterol (AUC 0.80) performed better (P < 0.001). Evaluation of scores based on the metabolic syndrome as defined by the National Cholesterol Education Program or with slight variations showed AUCs of 0.75 and 0.78, respectively. Rules based on all these approaches, while identifying 20-56% of the sample as screen positive, achieved sensitivities of 40-87% and specificities of 50-86%. CONCLUSIONS: Rules derived from clinical information, alone or combined with simple laboratory measures, can characterize degrees of diabetes risk in middle-aged adults, permitting preventive actions of appropriate intensity. Rules based on the metabolic syndrome are reasonable alternatives to rules derived from risk functions. PMID- 16043748 TI - Metabolic syndrome in type 1 diabetes: association with diabetic nephropathy and glycemic control (the FinnDiane study). AB - OBJECTIVE: The aim of this study was to estimate the prevalence of the metabolic syndrome in Finnish type 1 diabetic patients and to assess whether it is associated with diabetic nephropathy or poor glycemic control. RESEARCH DESIGN AND METHODS: In all, 2,415 type 1 diabetic patients (51% men, mean age 37 years, duration of diabetes 22 years) participating in the nationwide, multicenter Finnish Diabetic Nephropathy (FinnDiane) study were included. Metabolic syndrome was defined according to the National Cholesterol Education Program diagnostic criteria. Patients were classified as having normal albumin excretion rate (AER) (n = 1,261), microalbuminuria (n = 326), macroalbuminuria (n = 383), or end-stage renal disease (ESRD) (n = 164). Glycemic control was classified as good (HbA1c <7.5%), intermediate (7.5-9.0%), or poor (>9.0%). Creatinine clearance was estimated with the Cockcroft-Gault formula. RESULTS: The overall prevalence of metabolic syndrome was 38% in men and 40% in women. The prevalence was 28% in those with normal AER, 44% in microalbuminuric patients, 62% in macroalbuminuric patients, and 68% in patients with ESRD (P < 0.001). Patients with metabolic syndrome had a 3.75-fold odds ratio for diabetic nephropathy (95% CI 2.89-4.85), and all of the separate components of the syndrome were independently associated with diabetic nephropathy. The prevalence of metabolic syndrome was 31% in patients with good glycemic control, 36% in patients with intermediate glycemic control, and 51% in patients with poor glycemic control (P < 0.001). Similarly, metabolic syndrome increased with worsening creatinine clearance. CONCLUSIONS: The metabolic syndrome is a frequent finding in type 1 diabetes and increases with advanced diabetic nephropathy and worse glycemic control. PMID- 16043749 TI - Impaired absorption of insulin aspart from lipohypertrophic injection sites. PMID- 16043750 TI - Glucose Pulse: a simple method to estimate the amount of glucose oxidized during exercise in type 1 diabetic patients. PMID- 16043751 TI - Influenza vaccination among diabetic adults: related factors and trend from 1993 to 2001 in Spain. PMID- 16043752 TI - Diabetes self-management profile for flexible insulin regimens: cross-sectional and longitudinal analysis of psychometric properties in a pediatric sample. PMID- 16043753 TI - Plasma interleukin-10 concentration is positively related to insulin sensitivity in young healthy individuals. PMID- 16043754 TI - Clinical significance of urinary liver-type fatty acid-binding protein in patients with diabetic nephropathy. PMID- 16043755 TI - A paradoxical diurnal movement pattern in obese subjects with type 2 diabetes: a contributor to impaired appetite and glycemic control? PMID- 16043756 TI - Body iron stores are increased in overweight and obese women with polycystic ovary syndrome. PMID- 16043757 TI - Electrocardiographic QT interval prolongation and risk of primary cardiac arrest in diabetic patients. PMID- 16043758 TI - Endothelial dysfunction is correlated with microalbuminuria in children with short-duration type 1 diabetes. PMID- 16043759 TI - Association of erectile dysfunction with cardiovascular risk factors and increasing existing vascular disease in male chinese type 2 diabetic patients. PMID- 16043760 TI - Complications of Type 2 Diabetes Among Aboriginal Canadians: prevalence and associated risk factors. PMID- 16043761 TI - Expression of AdipoR1 in vivo in skeletal muscle is independently associated with measures of truncal obesity in middle-aged caucasian men. PMID- 16043762 TI - Prevalence of body iron excess in the metabolic syndrome. PMID- 16043763 TI - Association between cigarette smoking and metabolic syndrome: the Korea National Health and Nutrition Examination Survey. PMID- 16043764 TI - Antibodies to neuronal structures: innocent bystanders or neurotoxins? PMID- 16043765 TI - Second World Congress on the Insulin Resistance Syndrome: hypertension, cardiovascular disease, and treatment approaches. PMID- 16043766 TI - Transcutaneous gases determination in diabetic critical limb ischemia. PMID- 16043767 TI - Characteristics of California children with single versus multiple diabetic ketoacidosis hospitalizations (1998-2000). PMID- 16043768 TI - Statin neuropathy masquerading as diabetic autoimmune polyneuropathy. PMID- 16043769 TI - Use of glargine insulin before and during pregnancy in a woman with type 1 diabetes and Addison's Disease. PMID- 16043770 TI - Acute presentation of fetal hypertrophic cardiomyopathy in a type 1 diabetic pregnancy. PMID- 16043771 TI - The origin of subconductance levels in voltage-gated K+ channels. PMID- 16043772 TI - K channel subconductance levels result from heteromeric pore conformations. AB - Voltage-gated K channels assemble from four identical subunits symmetrically arranged around a central permeation pathway. Each subunit harbors a voltage sensing domain. The sigmoidal nature of the activation kinetics suggests that multiple sensors need to undergo a conformational change before the channel can open. Following activation, individual K channels alternate stochastically between two main permeation states, open and closed. This binary character of single channel behavior suggests the presence of a structure in the permeation pathway that can exist in only two conformations. However, single channel analysis of drk1 (K(v)2.1) K channels demonstrated the existence of four additional, intermediate conductance levels. These short-lived subconductance levels are visited when the channel gate moves between the closed and fully open state. We have proposed that these sublevels arise from transient heteromeric pore conformations, in which some, but not all, subunits are in the "open" state. A minimal model based on this hypothesis relates specific subconductance states with the number of activated subunits (Chapman et al., 1997). To stringently test this hypothesis, we constructed a tandem dimer that links two K channel subunits with different activation thresholds. Activation of this dimer by strong depolarizations resulted in the characteristic binary open-close behavior. However, depolarizations to membrane potentials in between the activation thresholds of the two parents elicited highly unusual single channel gating, displaying frequent visits to two subconductance levels. The voltage dependence and kinetics of the small and large sublevels associate them with the activation of one and two subunits, respectively. The data therefore support the hypothesis that subconductance levels result from heteromeric pore conformations. In this model, both sensor movement and channel opening have a subunit basis and these processes are allosterically coupled. PMID- 16043773 TI - Probing the geometry of the inner vestibule of BK channels with sugars. AB - The geometry of the inner vestibule of BK channels was probed by examining the effects of different sugars in the intracellular solution on single-channel current amplitude (unitary current). Glycerol, glucose, and sucrose decreased unitary current through BK channels in a concentration- and size-dependent manner, in the order sucrose > glucose > glycerol, with outward currents being reduced more than inward currents. The fractional decrease of outward current was more directly related to the fractional hydrodynamic volume occupied by the sugars than to changes in osmolality. For concentrations of sugars < or =1 M, the i/V plots for outward currents in the presence and absence of sugar superimposed after scaling, and increasing K(+)(i) from 150 mM to 2 M increased the magnitudes of the i/V plots with little effect on the shape of the scaled curves. These observations suggest that sugars < or =1 M reduce outward currents mainly by entering the inner vestibule and reducing the movement of K(+) through the vestibule, rather than by limiting diffusion-controlled access of K(+) to the vestibule. With 2 M sucrose, the movement of K(+) into the inner vestibule became diffusion limited for 150 mM K(+)(i) and voltages > +100 mV. Increasing K(+)(i) then relieved the diffusion limitation. An estimate of the capture radius based on the 5 pA diffusion-limited current for channels without the ring of negative charge at the entrance to the inner vestibule was 2.2 A. Adding the radius of a hydrated K(+) (6-8 A) then gave an effective radius for the entrance to the inner vestibule of 8-10 A. Such a functionally wide entrance to the inner vestibule together with our observation that even small concentrations of sugar in the inner vestibule reduce unitary current suggest that a wide inner vestibule is required for the large conductance of BK channels. PMID- 16043774 TI - Evidence for sequential ion-binding loci along the inner pore of the IRK1 inward rectifier K+ channel. AB - Steep rectification in IRK1 (Kir2.1) inward-rectifier K(+) channels reflects strong voltage dependence (valence of approximately 5) of channel block by intracellular cationic blockers such as the polyamine spermine. The observed voltage dependence primarily results from displacement, by spermine, of up to five K(+) ions across the narrow K(+) selectivity filter, along which the transmembrane voltage drops steeply. Spermine first binds, with modest voltage dependence, at a shallow site where it encounters the innermost K(+) ion and impedes conduction. From there, spermine can proceed to a deeper site, displacing several more K(+) ions and thereby producing most of the observed voltage dependence. Since in the deeper blocked state the leading amine group of spermine reaches into the cavity region (internal to the selectivity filter) and interacts with residue D172, its trailing end is expected to be near M183. Here, we found that mutation M183A indeed affected the deeper blocked state, which supports the idea that spermine is located in the region lined by the M2 and not deep in the narrow K(+) selectivity filter. As to the shallower site whose location has been unknown, we note that in the crystal structure of homologous GIRK1 (Kir3.1), four aromatic side chains of F255, one from each of the four subunits, constrict the intracellular end of the pore to approximately 10 A. For technical simplicity, we used tetraethylammonium (TEA) as an initial probe to test whether the corresponding residue in IRK1, F254, forms the shallower site. We found that replacing the aromatic side chain with an aliphatic one not only lowered TEA affinity of the shallower site approximately 100-fold but also eliminated the associated voltage dependence and, furthermore, confirmed that similar effects occurred also for spermine. These results establish the evidence for physically separate, sequential ion-binding loci along the long inner pore of IRK1, and strongly suggest that the aromatic side chains of F254 underlie the likely innermost binding locus for both blocker and K(+) ions in the cytoplasmic pore. PMID- 16043775 TI - Rapid increase in plasma membrane chloride permeability during wound resealing in starfish oocytes. AB - Plasma membrane wound repair is an important but poorly understood process. We used femtosecond pulses from a Ti-Sapphire laser to make multiphoton excitation induced disruptions of the plasma membrane while monitoring the membrane potential and resistance. We observed two types of wounds that depolarized the plasma membrane. At threshold light levels, the membrane potential and resistance returned to prewound values within seconds; these wounds were not easily observed by light microscopy and resealed in the absence of extracellular Ca(2+). Higher light intensities create wounds that are easily visible by light microscopy and require extracellular Ca(2+) to reseal. Within a few seconds the membrane resistance is approximately 100-fold lower, while the membrane potential has depolarized from -80 to -30 mV and is now sensitive to the Cl(-) concentration but not to that of Na(+), K(+), or H(+). We suggest that the chloride sensitivity of the membrane potential, after wound resealing, is due to the fusion of chloride-permeable intracellular membranes with the plasma membrane. PMID- 16043776 TI - Resting potential-dependent regulation of the voltage sensitivity of sodium channel gating in rat skeletal muscle in vivo. AB - Normal muscle has a resting potential of -85 mV, but in a number of situations there is depolarization of the resting potential that alters excitability. To better understand the effect of resting potential on muscle excitability we attempted to accurately simulate excitability at both normal and depolarized resting potentials. To accurately simulate excitability we found that it was necessary to include a resting potential-dependent shift in the voltage dependence of sodium channel activation and fast inactivation. We recorded sodium currents from muscle fibers in vivo and found that prolonged changes in holding potential cause shifts in the voltage dependence of both activation and fast inactivation of sodium currents. We also found that altering the amplitude of the prepulse or test pulse produced differences in the voltage dependence of activation and inactivation respectively. Since only the Nav1.4 sodium channel isoform is present in significant quantity in adult skeletal muscle, this suggests that either there are multiple states of Nav1.4 that differ in their voltage dependence of gating or there is a distribution in the voltage dependence of gating of Nav1.4. Taken together, our data suggest that changes in resting potential toward more positive potentials favor states of Nav1.4 with depolarized voltage dependence of gating and thus shift voltage dependence of the sodium current. We propose that resting potential-induced shifts in the voltage dependence of sodium channel gating are essential to properly regulate muscle excitability in vivo. PMID- 16043777 TI - Skeletal muscle HIF-1alpha expression is dependent on muscle fiber type. AB - Oxygen homeostasis is an essential regulation system for cell energy production and survival. The oxygen-sensitive subunit alpha of the hypoxia inducible factor 1 (HIF-1) complex is a key protein of this system. In this work, we analyzed mouse and rat HIF-1alpha protein and mRNA expression in parallel to energetic metabolism variations within skeletal muscle. Two physiological situations were studied using HIF-1alpha-specific Western blotting and semiquantitative RT-PCR. First, we compared HIF-1alpha expression between the predominantly oxidative soleus muscle and three predominantly glycolytic muscles. Second, HIF-1alpha expression was assessed in an energy metabolism switch model that was based on muscle disuse. These two in vivo situations were compared with the in vitro HIF 1alpha induction by CoCl(2) treatment on C(2)C(12) mouse muscle cells. HIF-1alpha mRNA and protein levels were found to be constitutively higher in the more glycolytic muscles compared with the more oxidative muscles. Our results gave rise to the hypothesis that the oxygen homeostasis regulation system depends on the fiber type. PMID- 16043778 TI - Flashing carotids in aortic valve disease. PMID- 16043779 TI - Fragile X and company: finding the right diagnosis. PMID- 16043780 TI - The triumvirate of acute hypertension. PMID- 16043782 TI - Axonal Charcot-Marie-Tooth disease: the fog is slowly lifting! PMID- 16043783 TI - Metabolic syndrome and intracranial atherosclerosis: a new link? PMID- 16043784 TI - The variable phenotype of FXTAS: a common cause of "idiopathic" disorders. PMID- 16043785 TI - The spectrum of presentations of venous infarction caused by deep cerebral vein thrombosis. AB - The classic features of thrombosis of the deep cerebral venous system are severe dysfunction of the diencephalon, reflected by coma and disturbances of eye movements and pupillary reflexes, resulting in poor outcome. However, partial syndromes without a decrease in the level of consciousness or brainstem signs exist, which may lead to initial misdiagnoses. The spectrum of clinical symptoms reflects the degree of venous congestion, which depends not only on the extent of thrombosis in the deep veins but also on the territory of the involved vessels and the establishment of venous collaterals. For example, thrombosis of the internal cerebral veins with (partially) patent basal veins and sufficient collaterals may result in relatively mild symptoms. Deep cerebral venous system thrombosis is an underdiagnosed condition when symptoms are mild, even in the presence of a venous hemorrhagic congestion. Identification of venous obstruction has important therapeutic implications. The diagnosis should be strongly suspected if the patient is a young woman, if the lesion is within the basal ganglia or thalamus, and especially if it is bilateral. PMID- 16043786 TI - Clinical and electrophysiologic features of CMT2A with mutations in the mitofusin 2 gene. AB - BACKGROUND: Axonal neuropathy linked to the CMT2A locus was originally associated with a mutation in the KIF1B gene. However, mutations in this gene have not been described associated with any other CMT2A families. Recently, mutations in the MFN2 gene, encoding the mitochondrial GTPase mitofusin 2 (Mfn2), have been identified as causative of CMT2A in seven families. The authors report three additional CMT2A families associated with novel mutations in highly conserved regions of the Mfn2 GTPase domain. METHODS: The authors performed a standardized neuromuscular and nerve conduction examination, genotyped known CMT loci, and analyzed the MFN2 gene by direct sequencing in three pedigrees and 10 additional probands affected by axonal CMT. RESULTS: Sequencing of the MFN2 gene revealed a novel mutation in each family (c.818T>G, c.638T>C, and c.314C>T). The largest family demonstrated an age-independent variable expression such that approximately one quarter of individuals with the mutation presented with features mild enough as to remain occult even with electrophysiologic evaluation. CONCLUSION: These results confirm that the majority of cases of CMT linked to the CMT2A locus are due to MFN2 mutations. The phenotype is largely indistinguishable from KIF1B-related CMT and from CMT2E and CMT2F. At least in some families, as many as 25% of individuals with MFN2 mutations may be asymptomatic and have a normal electrophysiologic examination, although a detailed neuromuscular examination may suggest the trait. Given the frequency of MFN2 mutations among CMT2 probands (3/13, or 23%), genetic testing of CMT2 patients should begin with a screen of the MFN2 gene. PMID- 16043787 TI - Association between benign and malignant peripheral nerve sheath tumors in NF1. AB - OBJECTIVE: People with neurofibromatosis type 1 (NF1) have a 10% lifetime risk of developing a malignant peripheral nerve sheath tumor (MPNST). MPNSTs are often metastatic and are a frequent cause of death among people with NF1. Clinical evidence suggests that most MPNSTs in people with NF1 develop from preexisting plexiform neurofibromas. However, it is not known whether an individual's risk of developing an MPNST is associated with the burden of benign neurofibromas. The authors conducted a study to determine whether people with NF1 who have benign neurofibromas of various kinds are at greater risk of developing MPNSTs than patients with NF1 who lack these benign tumors. METHODS: Clinical information on 476 NF1 probands in the Henri Mondor Database was analyzed by logistic regression to examine associations between MPNSTs and internal plexiform, superficial plexiform, subcutaneous, and cutaneous neurofibromas. RESULTS: Individuals with subcutaneous neurofibromas were approximately three times more likely to have internal plexiform neurofibromas or MPNSTs than individuals without subcutaneous neurofibromas. Individuals with internal plexiform neurofibromas were 20 times more likely to have MPNSTs than individuals without internal plexiform neurofibromas. When this analysis was done with both subcutaneous and internal plexiform neurofibromas as explanatory variables, only the association of MPNSTs with internal plexiform neurofibromas remained significant. CONCLUSIONS: The observation that malignant peripheral nerve sheath tumors are strongly associated with internal plexiform neurofibromas suggests that patients with neurofibromatosis type 1 with these benign tumors warrant increased surveillance for malignancy. PMID- 16043788 TI - Epileptic seizures during follow-up of patients treated for primary brain tumors. AB - OBJECTIVE: To determine the presentation, incidence, and severity of seizures in follow-up of patients treated for primary brain tumors. METHODS: A total of 234 consecutive patients attending an outpatient clinic for chemotherapy of a supratentorial brain tumor were examined. RESULTS: Seizures occurred in 183 patients. All patients with epilepsy were on antiepileptic drugs (AEDs). Compared with patients without epilepsy, patients with epilepsy had a higher proportion of low-grade gliomas (p < 0.001) and cortical tumor location (p < 0.001). In 158 (86.4%) patients, seizures were an early manifestation of the disease, and epilepsy developed in only 25 (13.6%) individuals in the course of the malignant disease. Generalization occurred in 50% of early seizures, but in only 19.1% of patients with seizures persisting after the initiation of AEDs and specific antitumor therapies. The reduction in seizure generalization was significant (p = 0.001). Despite AED and various antitumor treatments, one-half of the patients had a seizure within 1 month and two-thirds within 3 months before the last evaluation. CONCLUSIONS: Most tumor-related seizures first appear early in the course of disease, usually as a presenting manifestation. Antiepileptic drugs combined with specific antitumor treatments significantly reduce the rate of seizure generalization. However, most patients continue to have focal epilepsy during follow-up. PMID- 16043789 TI - Causes of death in remote symptomatic epilepsy. AB - OBJECTIVE: To determine the causes of death of individuals with developmental disabilities that occur more frequently among those with remote symptomatic epilepsy (i.e., epilepsy occurring in persons with developmental delay or identified brain lesions) than for those without. METHODS: The authors compared causes of mortality in persons with (n = 10,030) and without (n = 96,163) history of epilepsy in a California population of persons with mild developmental disabilities, 1988 to 2002. Subjects had traumatic brain injury, cerebral palsy, Down syndrome, autism, or a developmental disability with other or unknown etiology. There were 721,759 person-years of data, with 2,397 deaths. Underlying causes of death were determined from the State of California's official mortality records. Cause-specific death rates and standardized mortality ratios (SMRs) were computed for those with and without epilepsy relative to subjects in the California general population. Comparisons were then made between SMRs of those with and without epilepsy, and CIs on the ratios of SMRs were determined. RESULTS: Death rates for persons with epilepsy were elevated for several causes. The greatest excess was due to seizures (International Classification of Diseases 9 [ICD-9] 345; SMR 53.1, 95% CI 28.0 to 101.0) and convulsions (ICD-9 780.3; SMR 25.2, 95% CI 11.7 to 54.2). Other causes occurring more frequently in those with epilepsy included brain cancer (SMR 5.2, 95% CI 2.2 to 12.1), respiratory diseases (SMR 1.7, 95% CI 1.2 to 2.5), circulatory diseases (SMR 1.3, 95% CI 1.0 to 1.7), and accidents (SMR 2.7, 95% CI 1.9 to 3.7), especially accidental drowning (SMR 12.8, 95% CI 7.0 to 23.2). CONCLUSIONS: Remote symptomatic epilepsy is associated with an increased risk of death. Seizures, aspiration pneumonia, and accidental drowning are among the leading contributors. PMID- 16043790 TI - Progression in temporal lobe epilepsy: differential atrophy in mesial temporal structures. AB - OBJECTIVE: To determine the relationship between hippocampal, amygdalar, and entorhinal cortex atrophy and duration of epilepsy, presence of secondary generalized seizures, and prolonged childhood febrile convulsions in patients with pharmacologically intractable temporal lobe epilepsy (TLE). METHODS: Volumetric MRI of the hippocampus, amygdala, and entorhinal cortex were performed in 86 consecutive patients with TLE and 44 age- and sex-matched healthy control subjects. Linear regression analysis was used to explore the relation between the volumetric measurements and the clinical parameters. RESULTS: In simple regressions, duration of epilepsy but not age at seizure onset was related to hippocampal (r2 = 0.19, p < 0.001), entorhinal cortex (r2 = 0.11, p = 0.002), and amygdalar (r2 = 0.15, p < 0.001) atrophy ipsilateral to the seizure focus. Prediction of the regression function to time of onset of recurrent seizures (time = 0) resulted in a y intercept of < 0 for the hippocampus and the entorhinal cortex but was not different from 0 for the amygdala. Patients with a positive history of febrile convulsions had smaller hippocampal volumes ipsilateral to the seizure (p < 0.001). No relationship was found between febrile convulsions and entorhinal cortex and amygdalar volumes or between secondary generalization of seizures and any mesial temporal volume. CONCLUSION: Progressive volume loss in the mesial temporal lobe in relation to duration of epilepsy is not limited to the hippocampus but affects the entorhinal cortex and the amygdala. PMID- 16043791 TI - Neurocysticercosis: association between seizures, serology, and brain CT in rural Peru. AB - BACKGROUND: Neurocysticercosis (NCC) is the commonest helminthic CNS infection and the main cause of adult-onset seizures in developing countries, also frequent in industrialized countries because of immigration from endemic zones. Although NCC is commonly seen in individuals with seizures in endemic areas, its role as a cause of epilepsy has been questioned on the basis of the poor methodology of published studies. OBJECTIVE: To determine, in a cysticercosis-endemic area of the northern Peruvian coast, the frequency of 1) epileptic seizures, 2) serum antibodies to Taenia solium, 3) NCC-compatible findings on brain CT, and 4) the associations between these variables. METHODS: A community-wide screening survey for possible seizure cases was performed using a validated questionnaire. Positive respondents were later examined in the field by neurologists. Seizure cases were categorized as single seizure, active epilepsy, or inactive epilepsy. Serology was performed for all consenting individuals using immunoblot. Noncontrast brain CT scans were performed in all individuals with seizures and two groups of control subjects without seizures (seropositive and seronegative). RESULTS: The screening survey was applied to 903 permanent residents. Most positive respondents (114/137 [83.2%]) were examined by neurologists. The overall prevalence of epilepsy was 32.1 per 1,000 and that of active epilepsy was 16.6 per 1,000. Seroprevalence was 24.2% (200/825). Seroprevalence was associated with seizures (odds ratio 2.14; p = 0.026). Brain CT abnormalities compatible with NCC were more frequent in individuals with seizures and in those seropositive. CONCLUSION: In this hyperendemic area, an important proportion of seizure cases are associated with neurocysticercosis as demonstrated by serology or brain CT. PMID- 16043792 TI - Infantile spasms and intellectual outcomes in children with tuberous sclerosis complex. AB - OBJECTIVE: To assess intellectual outcomes in a clinic-based population of patients with tuberous sclerosis complex (TSC) who also have a history of infantile spasms (IS) and to identify clinical risk factors for mental retardation in these patients. METHODS: This is a retrospective study of 50 patients with TSC and IS seen consecutively at the Massachusetts General Hospital Tuberous Sclerosis Comprehensive Clinic from December 2001 to October 2003. Data were obtained by chart review and by interview with patients' parents. RESULTS: Thirty-two (64%) of 50 patients with TSC with IS were found to have mental retardation (IQ or developmental quotient < 70). Three clinical variables showed an association with mental retardation: increased duration of IS from clinical onset to cessation (odds ratio [OR] per 1-month interval 1.09, 95% CI: 1.03 to 1.15, p = 0.004), increased time from treatment initiation until IS cessation (OR 1.07, 95% CI: 1.01 to 1.14, p = 0.020), and poor control of other seizures after IS (OR 17.76, 95% CI: 3.47 to 129.1, p = 0.00004). The following variables did not show an association with intellectual outcome: gender, initial seizure type, age at IS onset, or time from IS onset to treatment initiation. CONCLUSIONS: In patients with tuberous sclerosis complex who also have a history of infantile spasms (IS), the rate of mental retardation may be lower than previously reported. The risk of mental retardation increases significantly with prolonged duration of IS, prolonged time from treatment initiation until the cessation of IS, and poor control of subsequent seizures after IS. PMID- 16043793 TI - Epidemiology of restless legs syndrome in French adults: a nationwide survey: the INSTANT Study. AB - OBJECTIVE: To evaluate the prevalence, characteristics, and treatment of restless legs syndrome (RLS) in France. METHODS: In this population-based survey, face-to face home interviews were conducted among a random sample of 10,263 French adults. A French translation of the four features defined by the International RLS Study Group in 1995 was used to assess the prevalence of symptoms consistent with a diagnosis of RLS. Data on severity of symptoms and their management were also collected. RESULTS: The 12-month prevalence of RLS symptoms in the French adult population was estimated to be 8.5% (95% CI 8.0%, 9.0%), with a higher prevalence (p < 0.001) observed in women (10.8%) than in men (5.8%). Prevalence increases with age until 64 years and decreases thereafter in both sexes. Half of the identified subjects reported symptoms once a week at least. Symptoms were more severe in subjects reporting symptoms once a week at least compared to subjects with less frequent symptoms. In this group, half of the subjects reported a family history, the age at onset was earlier, and severity of symptoms higher. RLS had been previously diagnosed in only 5.3% of the subjects who reported previous medical diagnosis, and recommended RLS drug treatment was received by 3.4% of the 28.7% currently treated subjects. CONCLUSIONS: Restless legs syndrome (RLS) occurred in 10% of women and 5% of men. RLS prevalence decreases after the age of 64. RLS is often underdiagnosed and few subjects receive recommended RLS drug treatment. PMID- 16043794 TI - Characteristics of idiopathic REM sleep behavior disorder and that associated with MSA and PD. AB - OBJECTIVE: To compare the clinical and video-polysomnographic (VPSG) characteristics of idiopathic REM sleep behavior disorder (RBD) vs the RBD seen in multiple system atrophy (MSA) and Parkinson disease (PD). METHODS: Clinical features and VPSG measures were evaluated in 110 consecutive nondemented subjects (26 MSA, 45 PD, and 39 idiopathic RBD) free of psychoactive medications referred for suspected RBD to our sleep unit over a 5-year period, with extended follow-up (mean 26.9 +/- 21.3 months). RESULTS: Across the three groups studied, logistic regression analysis demonstrated that there were no differences in the quality of RBD symptoms (e.g., nature of unpleasant dream recall or behaviors witnessed by bed partners), most PSG variables, abnormal behaviors captured by VPSG, and clinical response to clonazepam. When compared to subjects with PD, however, patients with MSA had a significantly shorter duration of disease, a higher REM sleep without atonia percentage, a greater periodic leg movement index, and less total sleep time. Subjects with idiopathic RBD, as compared to those with either MSA or PD, were more often male, had greater self-reported clinical RBD severity, and were more often aware of their abnormal sleep behaviors. CONCLUSIONS: REM sleep behavior disorder (RBD)-related symptoms and neurophysiologic features are qualitatively similar in RBD subjects with the idiopathic form, multiple system atrophy (MSA), and Parkinson disease (PD). Polysomnographic abnormalities associated with RBD in the setting of MSA are greater than in PD, suggesting a more severe dysfunction in the structures that modulate REM sleep. PMID- 16043795 TI - Lipid-lowering agent use at ischemic stroke onset is associated with decreased mortality. AB - BACKGROUND: 3-Hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors (statins), the most frequently used lipid-lowering agents (LLAs) have neuroprotective effects in rodent models of ischemic stroke. The authors hypothesized that patients with ischemic stroke taking LLAs would have better outcomes than patients not taking LLAs. METHODS: The Northern Manhattan Study is a population based study designed to determine stroke incidence and prognosis in a multiethnic, urban population. Northern Manhattan residents age 40 years or older diagnosed with their first ischemic stroke were eligible. Patients or their proxies were interviewed regarding medications being taken at home before stroke onset. The NIH Stroke Scale was used to assess stroke severity, categorized as mild (< or =5), moderate (6 to 13), or severe (> or =14), and the Barthel Index at 6 months to assess functional outcome. Clinical worsening in hospital was recorded by trial neurologists. Odds ratios and 95% CIs for association of LLA use and stroke severity, mortality, and functional outcome were calculated using logistic regression. RESULTS: Of 650 patients, 57 (8.8%) were taking LLAs. The majority (90.9%) of LLA users were taking a statin. Clinical worsening in hospital occurred less frequently among patients taking LLAs at stroke onset (6.3% vs 18.2%; p = 0.04). Ninety-day mortality was lower in those taking LLAs (1.8% vs 10.6%, p = 0.03). The proportion of patients with severe stroke among those taking LLAs was not lower (10.7% vs 16.8%, p = 0.39). CONCLUSION: Patients taking lipid-lowering agents (LLAs) at the time of an ischemic stroke may have lower poststroke mortality and a lower risk of worsening during hospitalization. Prospective studies are warranted to determine whether LLAs, and statins in particular, have neuroprotective properties or other beneficial effects in acute ischemic stroke. PMID- 16043796 TI - APOE, vascular pathology, and the AD brain. AB - OBJECTIVE: To use neuropathologic data to examine the association between APOE genotype and cerebrovascular lesions commonly found in Alzheimer disease (AD), as well as neuritic senile plaque (SP) and neurofibrillary tangle (NFT) burden. METHODS: The sample comprised brains from 96 men and 3 women who fulfilled NIA Reagan criteria for intermediate to high likelihood of AD. Region-specific and global measures of gross cerebrovascular disease, arteriolosclerosis, white matter lesions, microinfarcts, amyloid angiopathy, neuritic SP, and NFT burden were compared among those who had at least one APOE-epsilon4 vs those who did not. Pairwise rank-order correlations between measures were calculated. The association between APOE epsilon4 status and measures of vascular and AD pathology, adjusting for age at death, sex, brain weight, and Braak stage, were evaluated. RESULTS: APOE-epsilon4 was not associated with gross cerebrovascular pathology. Compared to those who were negative, brains from epsilon4 individuals had a greater degree of small vessel arteriolosclerosis (p = 0.04) and perivascular macrophage infiltration (p = 0.06), but not other markers of small vessel disease or white matter myelin loss. Microinfarcts in the deep nuclei were associated with epsilon4 (p = 0.009), whereas cortical and subcortical microinfarcts were not. There was a trend toward association between APOE genotype and amyloid angiopathy (p = 0.08), and epsilon4 was associated with neuritic SP burden, but not NFT. CONCLUSION: APOE-epsilon4 is associated with small vessel arteriolosclerosis, microinfarcts of the deep nuclei, neuritic senile plaque density, and amyloid angiopathy in patients with autopsy-proven Alzheimer disease (AD). These results suggest a role for epsilon4 in some of the microvascular changes commonly found in AD and are consistent with a potential amyloidogenic role for epsilon4. PMID- 16043797 TI - Episodic and semantic memory tasks activate different brain regions in Alzheimer disease. AB - OBJECTIVE: To compare brain activity identified by fMRI in subjects with Alzheimer disease (AD) and older healthy controls (HCs) performing an episodic/working memory (EWM) and semantic memory (SM) task. METHODS: Nine AD (mean age 73.6) and 10 HC (mean age 71.8) subjects underwent an fMRI memory paradigm. Tasks comprised 1) baseline (recognizing a single digit presented for 1 second), 2) SM (addition of two single digits, always producing a single digit answer), and 3) EWM (recall of the previous single digit on the stimulus of the next digit). Each condition was presented in 2-minute blocks with a shorter and longer time interval for the first and second minute within blocks. RESULTS: Comparing AD and HC subjects, there were no activated brain regions in common for EWM > SM, but left anterior cingulate (Brodmann area [BA] 24, 0, 31, 4) and left medial frontal lobe gyrus (BA 25, -6, 23, -15) were activated by both groups for SM > EWM. Key differences were that for EWM > SM, HC subjects activated the right parahippocampal gyrus, whereas subjects with AD activated the right superior frontal gyrus and left uncus. CONCLUSIONS: Subjects with Alzheimer disease (AD) recruited brain regions for easier episodic/working memory (EWM) tasks used by healthy controls (HCs) for more difficult EWM tasks. AD subjects recruited brain regions for semantic memory tasks used by HCs for more difficult EWM tasks. The authors propose a functional "memory reserve" model of compensatory recruitment according to task difficulty and underlying neuropathology. PMID- 16043798 TI - Impaired spatial coding within objects but not between objects in prosopagnosia. AB - BACKGROUND: Patients with prosopagnosia from occipitotemporal lesions have impaired perception of the configuration of facial features. This may be an example of impaired "within-object" spatial coding, which others propose to be distinct from "between-object" spatial coding. OBJECTIVE: To determine whether the prosopagnosic deficit in perceiving spatial configuration was specific to within-face and not between-face spatial coding and whether the deficit was face selective or extended to objects other than faces. METHODS: Six prosopagnosic patients were tested using an oddity paradigm in which they detected which of three simultaneously seen stimuli was an altered target. In the "within-face" task, the target face had altered interocular distance or mouth position. In the "between-face" task, the target face was located farther away from the other two. In the "within-object" task, the stimulus was a two-dot pattern, and the target pattern had altered interdot distance. RESULTS: Spatial judgments were impaired within faces for all six patients and within the two-dot pattern for five of six patients. However, all six had normal between-face spatial perception. CONCLUSIONS: Impaired perception of spatial relations in prosopagnosia is selective to the spatial structure within individual objects and spares the perception of spatial location of objects. It is not specific to faces. It reveals a process involved in analyzing object structure, consistent with the patients' deficits in recognizing facial identity, and illustrates a different type of "visuospatial" defect. PMID- 16043799 TI - A clinical rating scale for Batten disease: reliable and relevant for clinical trials. AB - BACKGROUND: Batten disease (juvenile neuronal ceroid lipofuscinosis [JNCL]) is an autosomal recessive neurodegenerative disorder characterized by blindness, seizures, and relentless decline in cognitive, motor, and behavioral function. Onset is in the early school years, with progression to death typically by late adolescence. Development of a clinical instrument to quantify severity of illness is a prerequisite to eventual assessment of experimental therapeutic interventions. OBJECTIVE: To develop a clinical rating instrument to assess motor, behavioral, and functional capability in JNCL. METHODS: A clinical rating instrument, the Unified Batten Disease Rating Scale (UBDRS), was developed by the authors to assess motor, behavioral, and functional capability in JNCL. Children with verified JNCL were evaluated independently by three neurologists. Intraclass correlation coefficients (ICCs) were used to estimate the interrater reliability for total scores in each domain. Interrater reliability for scale items was assessed with weighted kappa statistics. RESULTS: Thirty-one children with confirmed JNCL (10 boys, 21 girls) were evaluated. The mean age at symptom onset was 6.1 +/- 1.6 years, and the mean duration of illness was 9.0 +/- 4.4 years. The ICCs for the domains were as follows: motor = 0.83, behavioral = 0.68, and functional capability = 0.85. CONCLUSIONS: The Unified Batten Disease Rating Scale (UBDRS) is a reliable instrument that effectively tests for neurologic function in blind and demented patients. In its current form, the UBDRS is useful for monitoring the diverse clinical findings seen in Batten disease. PMID- 16043800 TI - Brain atrophy and lesion load in a large population of patients with multiple sclerosis. AB - OBJECTIVE: To measure white matter (WM) and gray matter (GM) atrophy and lesion load in a large population of patients with multiple sclerosis (MS) using a fully automated, operator-independent, multiparametric segmentation method. METHODS: The study population consisted of 597 patients with MS and 104 control subjects. The MRI parameters were abnormal WM fraction (AWM-f), global WM-f (gWM-f), and GM fraction (GM-f). RESULTS: Significant differences between patients with MS and control subjects included higher AWM-f and reduced gWM-f and GM-f. MRI data showed significant differences between patients with relapsing-remitting and secondary progressive forms of MS. Significant correlations between MRI parameters and between MRI and clinical data were found. CONCLUSIONS: Patients with multiple sclerosis have significant atrophy of both white matter (WM) and gray matter (GM); secondary progressive patients have significantly more atrophy of both WM and GM than do relapsing-remitting patients and a significantly higher lesion load (abnormal WM fraction); lesion load is related to both WM and even more to GM atrophy; lesion load and WM and GM atrophy are significantly related to Expanded Disability Status Scale score and age at onset (suggesting that the younger the age at disease onset, the worse the lesion load and brain atrophy); and GM atrophy is the most significant MRI variable in determining the final disability. PMID- 16043801 TI - Ethyl-EPA in Huntington disease: a double-blind, randomized, placebo-controlled trial. AB - BACKGROUND: Preliminary evidence suggests beneficial effects of pure ethyl eicosapentaenoate (ethyl-EPA) in Huntington disease (HD). METHODS: A total of 135 patients with HD were randomized to enter a multicenter, double-blind, placebo controlled trial on the efficacy of 2 g/d ethyl-EPA vs placebo. The Unified Huntington's Disease Rating Scale (UHDRS) was used for assessment. The primary end point was outcome at 12 months on the Total Motor Score 4 subscale (TMS-4). Analysis of covariance (ANCOVA) and a chi2 test on response, defined as absence of increase in the TMS-4, were performed. RESULTS: A total of 121 patients completed 12 months, and 83 did so without protocol violations (PP cohort). Intent-to-treat (ITT) analysis revealed no significant difference between ethyl EPA and placebo for TMS-4. In the PP cohort, ethyl-EPA proved better than placebo on the chi2 test on TMS-4 (p < 0.05), but missed significance on ANCOVA (p = 0.06). Secondary end points (ITT cohort) showed no benefit of ethyl-EPA but a significantly worse outcome in the behavioral severity and frequency compared with placebo. Exploring moderators of the efficacy of ethyl-EPA on TMS-4 showed a significant interaction between treatment and a factor defining patients with high vs low CAG repeats. Reported adverse events were distributed equally between treatment arms. CONCLUSIONS: Ethyl-eicosapentaenoate (ethyl-EPA) (purity > 95%) had no benefit in the intent-to-treat cohort of patients with Huntington disease, but exploratory analysis revealed that a significantly higher number of patients in the per protocol cohort, treated with ethyl-EPA, showed stable or improved motor function. Further studies of the potential efficacy of ethyl-EPA are warranted. PMID- 16043802 TI - The smoking-thrombolysis paradox and acute ischemic stroke. AB - Smokers with acute myocardial infarction have better outcomes after thrombolysis than nonsmokers. The authors evaluated the independent effect of smoking on short term outcome following IV thrombolysis for acute ischemic stroke. After adjusting for covariates, recent smokers who received thrombolysis had a significantly greater drop in 24-hour median stroke severity scores from baseline than nonsmokers who received thrombolysis and lower mortality over 1 year. PMID- 16043803 TI - Association of the metabolic syndrome with intracranial atherosclerotic stroke. AB - To investigate the impact of metabolic syndrome (MetSD) on the development of intracranial atherosclerotic stroke, the authors evaluated the components of the MetSD in 512 patients with stroke. The MetSD was observed most frequently in patients with intracranial atherosclerosis (p = 0.007). In multiple regression analysis, the MetSD, but not conventional risk factors, was independently associated with intracranial atherosclerosis (p = 0.005). The results suggest that treatment of metabolic abnormalities may be an important prevention strategy for intracranial atherosclerosis. PMID- 16043804 TI - Initial diagnoses given to persons with the fragile X associated tremor/ataxia syndrome (FXTAS). AB - Fragile X-associated tremor/ataxia syndrome (FXTAS) is a newly described disorder that occurs in premutation carriers of the fragile X mental retardation 1 (FMR1) gene. Fifty-six patients with FXTAS were given 98 prior diagnoses: most were in the categories of parkinsonism, tremor, ataxia, dementia, or stroke. Data from this study and others were used to develop guidelines for FMR1 diagnostic testing for FXTAS. PMID- 16043805 TI - Acute painful neuropathy in thallium poisoning. AB - Dysesthesia, allodynia, distal muscle weakness, and sensory impairment were noted in two patients with acute thallium intoxication. Two months later, nerve conduction studies showed an axonal degeneration. Sural nerve biopsy disclosed a decreased fiber density in the large myelinated fibers. Quantitative sensory testing also revealed an impairment of pinprick, temperature, and touch sensations. Cutaneous nerve biopsy confirmed a loss of epidermal nerves indicating an involvement of the small sensory nerves. PMID- 16043806 TI - Ultrasonography shows extensive nerve enlargements in multifocal motor neuropathy. AB - Using ultrasonography we found multiple sites with nerve enlargement along the course of the brachial plexus, median, ulnar, and radial nerves in the majority of 21 patients with multifocal motor neuropathy. Sonography and electrophysiologic studies showed more abnormalities than expected on purely clinical grounds. Moreover, sonography revealed nerve enlargement without clinical or electrophysiologic abnormalities. PMID- 16043807 TI - New calcium channel mutations predict aberrant RNA splicing in episodic ataxia. AB - Episodic ataxia type 2 (EA2) is an autosomal dominant channelopathy characterized by paroxysmal cerebellar ataxia. Previous studies suggest that most EA2 cases are associated with mutations in the alpha1A subunit of the P/Q-type voltage-gated calcium channel gene CACNA1A. In a UK national study, the authors analyzed 15 index cases with typical EA2 and identified two unreported intronic mutations that predict aberrant splicing. PMID- 16043808 TI - Mechanical hyperalgesia in complex regional pain syndrome: a role for TNF-alpha? AB - Plasma concentrations of soluble tumor necrosis factor alpha (TNF-alpha) receptor type I (sTNF-RI) were assessed in two complex regional pain syndrome (CRPS) patient groups (n = 30 and n = 16) and healthy controls (n = 25). Patients with CRPS and mechanical hyperalgesia had higher levels of sTNF-RI (1,661.8 +/- 146.8 pg/mL) compared with those with CRPS with identical clinical appearance but without hyperalgesia (1,155.9 +/- 56.3 pg/mL) and controls (1,239.5 +/- 42.9 pg/mL). This study suggests involvement of TNF-alpha in mechanical hyperalgesia of CRPS. PMID- 16043809 TI - Clinical features of status epilepticus in patients with HIV infection. AB - The authors reviewed the records of 42 patients with HIV infection and status epilepticus (SE). Brain tumor and infection were the most common etiologies. The median duration of SE was 2.0 +/- 10 hours. Most patients (37 [88%]) responded to IV benzodiazepine or phenytoin treatment. Nevertheless, 12 (29%) patients died and 15 (36%) developed new neurologic deficits. In patients with HIV infection, aggressive management of seizures may limit the risk of SE. PMID- 16043810 TI - Vagus nerve stimulation for epilepsy: randomized comparison of three stimulation paradigms. AB - Vagus nerve stimulation (VNS) is an effective adjunctive treatment for intractable epilepsy. However, the optimal range of device duty-cycles [on/(on + off times)] is poorly understood. The authors performed a multicenter, randomized trial of three unique modes of VNS, which varied primarily by duty-cycle. The results indicate that the three duty-cycles were equally effective. The data support the use of standard duty-cycles as initial therapy. PMID- 16043811 TI - The prognosis of hearing impairment complicating HIV-negative cryptococcal meningitis. AB - Eight patients who had sensorineural hearing loss (SNHL) associated with cryptococcal meningitis were studied. After a minimum 3-year follow-up, one had died. Among the seven survivors, three had improved, two stabilized, and two progressed. Predictive factors included visual disturbance, meningeal enhancements on MRI, and a CSF cryptococcal antigen titer of >1:1,024. SNHL accounted for 30.8% (8/26) of cryptococcal meningitis patients in our study. PMID- 16043812 TI - Familial Alzheimer disease: decreases in CSF Abeta42 levels precede cognitive decline. AB - CSF amyloid beta-peptide 42 (Abeta42) levels in presymptomatic subjects with pathogenic mutations in the PS1 gene are significantly lower than in an age matched control group. Consequently, in these subjects, there is a window of opportunity estimated as at least 4 to 12 years to evaluate the ability of any putative prophylactic therapy to decrease, arrest, or reverse abnormalities in Abeta42 metabolism many years before clinical symptoms of Alzheimer disease are otherwise likely to occur. PMID- 16043813 TI - A critical period for the impact of amygdala damage on the emotional enhancement of memory? AB - The amygdala is crucial in modulating enhanced memory for emotionally arousing material. The authors provide evidence that unilateral lesions of the human amygdala arising early in development, but not in adulthood, are associated with a loss of the expected superior retrieval of emotionally arousing over neutral material. This adds to evidence for an early critical period in the development of amygdala function. PMID- 16043814 TI - Creutzfeldt-Jakob disease mimicking radiologic posterior reversible leukoencephalopathy. PMID- 16043815 TI - Creutzfeldt-Jakob disease manifesting as posterior cortical dementia. PMID- 16043816 TI - Fragile X-associated tremor/ataxia syndrome presenting in a woman after chemotherapy. PMID- 16043817 TI - Systemic mastocytosis: a potential neurologic emergency. PMID- 16043818 TI - Neurologic oral manifestations caused by a new formulation of mirtazapine. PMID- 16043819 TI - Personal history: The carefree realm of sleep. PMID- 16043820 TI - Sirolimus may not cause neurotoxicity in kidney and liver transplant recipients. PMID- 16043821 TI - Expert medical testimony: responsibilities of medical societies. PMID- 16043822 TI - Benefits of cognitive-motor intervention in MCI and mild to moderate Alzheimer disease. PMID- 16043823 TI - Auditory agnosia caused by a tectal germinoma. PMID- 16043824 TI - Analysis of leaf proteome after UV-B irradiation in maize lines differing in sensitivity. AB - UV-B radiation causes diverse morphological and physiological responses in plants, but the underlying mechanisms governing these integrated responses are unknown. In this study, we systematically surveyed responses of maize leaves to UV-B radiation using DIGE 2D gels and identified selected proteins by mass spectrometry and immunodetection analysis. To identify changes in protein accumulation in response to UV-B radiation, a line (b, pl W23) deficient in flavonoid sunscreen compounds and hence similar to commercial corn was used. In addition, its proteome in natural UV-B conditions was compared with that of two maize landraces from high altitudes (Cacahuacintle and Confite Puneno) that have improved UV-B tolerance. Protein patterns in adult maize leaves (Zea mays) were documented after growth for 21 days in sunlight depleted of UV-B radiation or growth in sunlight including an 8-h UV-B supplementation during 1 day in the field. We found that there is a very high correlation between previously documented mRNA accumulation assessed by microarray hybridization and quantitative real time reverse transcription-PCR and protein expression after UV B irradiation in leaves of W23. Multiple isoforms were confirmed for some proteins; at least one protein, pyruvate, phosphate dikinase, is regulated post translationally by phosphorylation by UV-B exposure. Proteins differentially regulated by UV-B radiation in W23 with higher levels under similar UV-B conditions in high altitude plants were also identified. These could be genetically fixed traits conferring UV-B tolerance and offer clues to specific adaptations to living in high ambient UV-B conditions. PMID- 16043825 TI - Epidermal growth factor receptor tyrosine kinase inhibitors plus chemotherapy: case closed or is the jury still out? PMID- 16043826 TI - A teachable moment for oncologists: cancer survivors, 10 million strong and growing! PMID- 16043827 TI - Unrelated donor marrow transplantation for B-cell chronic lymphocytic leukemia after using myeloablative conditioning: results from the Center for International Blood and Marrow Transplant research. AB - PURPOSE: To determine the role of myeloablative conditioning and unrelated donor (URD) bone marrow transplantation in the treatment of patients with advanced B cell chronic lymphocytic leukemia (CLL). PATIENTS AND METHODS: A total of 38 CLL patients received a matched URD transplant using bone marrow procured by the National Marrow Donor Program. The median age was 45 years (range, 26 to 57 years), the median time from diagnosis was 51 months, and the median number of prior chemotherapy regimens was three. Fifty-five percent of patients were chemotherapy refractory and 89% had received fludarabine. Conditioning included total-body irradiation in 92% of patients. Graft-versus-host disease (GVHD) prophylaxis consisted of methotrexate with cyclosporine or tacrolimus for 82% of patients. RESULTS: Twenty-one patients (58%) achieved complete response and six (17%) achieved partial response. Incidences of grades 2 to 4 acute GVHD were 45% at 100 days and incidences of chronic GVHD were 85% at 5 years. Eleven patients are alive and disease free at a median of 6 years (range, 3.0 to 9.0 years). Five year overall survival, failure-free survival, disease progression rates, and treatment-related mortality (TRM) were 33%, 30%, 32%, and 38% respectively. CONCLUSION: These data demonstrate that lasting remissions can be achieved after URD transplantation in patients with advanced CLL. High TRM suggest that myeloablative conditioning and HLA-mismatched donors should be avoided in future protocols, and it is mandatory to investigate transplant strategies with a lower morbidity and mortality, including the use of nonmyeloablative regimens. PMID- 16043828 TI - Mutations in the epidermal growth factor receptor and in KRAS are predictive and prognostic indicators in patients with non-small-cell lung cancer treated with chemotherapy alone and in combination with erlotinib. AB - PURPOSE: Epidermal growth factor receptor (EGFR) mutations have been associated with tumor response to treatment with single-agent EGFR inhibitors in patients with relapsed non-small-cell lung cancer (NSCLC). The implications of EGFR mutations in patients treated with EGFR inhibitors plus first-line chemotherapy are unknown. KRAS is frequently activated in NSCLC. The relationship of KRAS mutations to outcome after EGFR inhibitor treatment has not been described. PATIENTS AND METHODS: Previously untreated patients with advanced NSCLC in the phase III TRIBUTE study who were randomly assigned to carboplatin and paclitaxel with erlotinib or placebo were assessed for survival, response, and time to progression (TTP). EGFR exons 18 through 21 and KRAS exon 2 were sequenced in tumors from 274 patients. Outcomes were correlated with EGFR and KRAS mutations in retrospective subset analyses. RESULTS: EGFR mutations were detected in 13% of tumors and were associated with longer survival, irrespective of treatment (P < .001). Among erlotinib-treated patients, EGFR mutations were associated with improved response rate (P < .05) and there was a trend toward an erlotinib benefit on TTP (P = .092), but not improved survival (P = .96). KRAS mutations (21% of tumors) were associated with significantly decreased TTP and survival in erlotinib plus chemotherapy-treated patients. CONCLUSION: EGFR mutations may be a positive prognostic factor for survival in advanced NSCLC patients treated with chemotherapy with or without erlotinib, and may predict greater likelihood of response. Patients with KRAS-mutant NSCLC showed poorer clinical outcomes when treated with erlotinib and chemotherapy. Further studies are needed to confirm the findings of this retrospective subset analysis. PMID- 16043829 TI - TRIBUTE: a phase III trial of erlotinib hydrochloride (OSI-774) combined with carboplatin and paclitaxel chemotherapy in advanced non-small-cell lung cancer. AB - PURPOSE: Erlotinib is a potent reversible HER1/epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor with single-agent activity in patients with non small-cell lung cancer (NSCLC). Erlotinib was combined with chemotherapy to determine if it could improve the outcome of patients with NSCLC. PATIENTS AND METHODS: TRIBUTE randomly assigned patients with good performance status and previously untreated advanced (stage IIIB/IV) NSCLC to erlotinib 150 mg/d or placebo combined with up to six cycles of carboplatin and paclitaxel, followed by maintenance monotherapy with erlotinib. Random assignment was stratified by stage, weight loss in the previous 6 months, measurable disease, and treatment center. The primary end point was overall survival (OS). Secondary end points included time to progression (TTP), objective response (OR), and duration of response. RESULTS: There were 1,059 assessable patients (526 erlotinib; 533 placebo). Median survival for patients treated with erlotinib was 10.6 v 10.5 months for placebo (hazard ratio, 0.99; 95% CI, 0.86 to 1.16; P = .95). There was no difference in OR or median TTP. Patients who reported never smoking (72 erlotinib; 44 placebo) experienced improved OS in the erlotinib arm (22.5 v 10.1 months for placebo), though no other prespecified factors showed an advantage in OS with erlotinib. Erlotinib and placebo arms were equivalent in adverse events (except rash and diarrhea). CONCLUSION: Erlotinib with concurrent carboplatin and paclitaxel did not confer a survival advantage over carboplatin and paclitaxel alone in patients with previously untreated advanced NSCLC. Never smokers treated with erlotinib and chemotherapy seemed to experience an improvement in survival and will undergo further investigation in future randomized trials. PMID- 16043831 TI - Preschool vision screening tests administered by nurse screeners compared with lay screeners in the vision in preschoolers study. AB - PURPOSE: To compare the performance of nurse screeners with that of lay screeners in administering preschool vision screening tests. METHODS: Trained nurse and lay screeners administered the Retinomax Autorefractor (Right Manufacturing, Virginia Beach, VA), SureSight Vision Screener (Welch Allyn, Inc., Skaneateles Falls, NY), crowded Linear Lea Symbols visual acuity (VA) test at 10 ft (Precision Vision, Inc., La Salle, IL), and Stereo Smile II test (Stereo Optical, Inc., Chicago, IL) to 3- to 5-year-old Head Start participants. Lay screeners also administered a crowded Single Lea Symbols VA test at 5 ft (Good-Lite, Inc.). Screening results were compared with the classification of the children according to the presence of one or more of four conditions (amblyopia, strabismus, significant refractive error, and unexplained reduced VA) based on the results of a gold standard eye examination by study-certified optometrists and ophthalmologists. The primary outcome measure was sensitivity for detecting children with one or more targeted conditions at 0.90 specificity. RESULTS: Nurse screeners achieved slightly higher sensitivities with the Retinomax, SureSight, and Stereo Smile II tests than did lay screeners; however, most differences were small and not statistically significant. Nurse screeners achieved significantly higher sensitivity with the Linear Lea Symbols VA test than did lay screeners. Lay screeners achieved strikingly higher sensitivity with the Single Lea Symbols VA test than did nurse or lay screeners using the Linear Lea Symbols VA test. Combining the Stereo Smile II test with each of the other tests did not result in improved sensitivities for detecting one or more targeted conditions. CONCLUSIONS: Nurse and lay screeners can achieve similar sensitivity, when specificity is set at 0.90, for detecting preschool children in need of a comprehensive eye examination. PMID- 16043830 TI - Riding the crest of the teachable moment: promoting long-term health after the diagnosis of cancer. AB - PURPOSE: Cancer survivors are at increased risk for several comorbid conditions, and many seek lifestyle change to reduce dysfunction and improve long-term health. To better understand the impact of cancer on adult survivors' health and health behaviors, a review was conducted to determine (1) prevalent physical health conditions, (2) persistent lifestyle changes, and (3) outcomes of previous lifestyle interventions aimed at improving health within this population. METHODS: Relevant studies from 1966 and beyond were identified through MEDLINE and PubMed searches. RESULTS: Cancer survivors are at increased risk for progressive disease but also for second primaries, osteoporosis, obesity, cardiovascular disease, diabetes, and functional decline. To improve overall health, survivors frequently initiate diet, exercise, and other lifestyle changes after diagnosis. However, those who are male, older, and less educated are less likely to adopt these changes. There also is selective uptake of messages, as evidenced by findings that only 25% to 42% of survivors consume adequate amounts of fruits and vegetables, and approximately 70% of breast and prostate cancer survivors are overweight or obese. Several behavioral interventions show promise for improving survivors' health-related outcomes. Oncologists can play a pivotal role in health promotion, yet only 20% provide such guidance. CONCLUSION: With 64% of cancer patients surviving > 5 years beyond diagnosis, oncologists are challenged to expand their focus from acute care to managing the long-term health consequences of cancer. Although more research is needed, opportunities exist for oncologists to promote lifestyle changes that may improve the length and quality of life of their patients. PMID- 16043833 TI - Mechanistic insights into glaucoma provided by experimental genetics the cogan lecture. PMID- 16043835 TI - Glaucoma: macrocosm to microcosm the Friedenwald lecture. PMID- 16043837 TI - The dark side of light: rhodopsin and the silent death of vision the proctor lecture. PMID- 16043839 TI - Retinoblastoma in the 20th century: past success and future challenges the Weisenfeld lecture. PMID- 16043840 TI - Lacrimal gland involvement in graft-versus-host disease: a murine model. AB - PURPOSE: To describe lacrimal gland involvement in a murine model of acute graft versus-host disease (GVHD). METHODS: Histopathologic examination was performed on lacrimal glands of mice affected by GVHD at 1, 2, 4, and 6 weeks after allogeneic bone marrow transplantation (BMT). Histopathologic scoring, based on characteristic GVHD findings in human disease involved evaluation of periductal inflammation, apoptosis, ductal stasis, ductal debris, and fibrosis. CD3, CD4, CD8, CD20, and CD68 antibodies were used to stain leukocyte subsets in GVHD lacrimal gland infiltrates. Lacrimal glands from syngeneic BMT mice were used in control experiments. RESULTS: Patchy periductal inflammation and focal fibrosis were significantly elevated as early as 2 weeks after allogeneic BMT. Histopathologic scoring of lacrimal glands after allogeneic BMT was significantly different at 4 (P = 0.005) and 6 (P < 0.0001) weeks when compared with scores in syngeneic control mice. The leukocytes in lacrimal gland GVHD infiltrates were predominately CD3+ T lymphocytes, most of which were CD8+, with fewer CD4+ cells present. CONCLUSIONS: This study describes the first murine model of lacrimal gland GVHD with features that closely mimic those described in human disease and indicates that lacrimal involvement occurs in acute GVHD. PMID- 16043841 TI - Shape of the retinal surface in emmetropia and myopia. AB - PURPOSE: To determine and compare the shapes of the retinas of emmetropic and myopic eyes. METHODS: Nonrotationally symmetrical ellipsoids were mathematically fitted to the retinal surfaces of 21 emmetropic and 66 myopic eyes (up to -12 D) of participants aged 18 to 36 years (mean, 25.5) using transverse axial and sagittal images derived from magnetic resonance imaging. RESULTS: The shapes of the ellipsoids varied considerably between subjects with similar refractive errors. The shapes were oblate (steepening toward the equator) in most of the emmetropic eyes (i.e., the axial dimensions of the ellipsoids were smaller than both the vertical and horizontal dimensions). As myopia increased, all ellipsoid dimensions increased with the axial dimension increasing more than the vertical dimension, which in turn increased more than the horizontal dimension (increases in approximate ratios 3:2:1). The relative difference in the increase of these dimensions meant that as the degree of myopia increased the retinal shape decreased in oblateness. However, few myopic eyes were prolate (flattening toward the equator). Independent of myopia, the ellipsoids were tilted about the vertical axis by 11 degrees +/- 13 degrees , and ellipsoid centers were decentered horizontally by 0.5 +/- 0.4 mm nasally and 0.2 +/- 0.5 mm inferiorly, relative to the fovea. CONCLUSIONS: In general both emmetropic and myopic retinas are oblate in shape, although myopic eyes less so. This finding may be relevant to theories implicating the peripheral retina in the development of myopia. PMID- 16043842 TI - Expression of the IGF system in normal and diabetic transgenic (mRen-2)27 rat eye. AB - PURPOSE: In the present study, a recently described model of diabetic eye disease was used to investigate the distribution of the insulin-like growth factor (IGF) system in the eyes of transgenic (mRen-2)27 rats (exhibiting hypertension and elevated serum and ocular renin levels) with streptozotocin-induced diabetes. METHODS: Female transgenic (mRen-2)27 rats were randomized to receive either streptozotocin (diabetic) or citrate buffer (control). After 10 months, the rats were killed and the eyes fixed and embedded in paraffin. In situ hybridization (ISH) was used to document the cellular distribution of mRNAs for components of the IGF system (IGF-I, IGF-I receptor [IGFIR] and IGF binding proteins [IGFBP]1 to -6) in the eyes. RESULTS: In nondiabetic rats, mRNA for IGFBP-1, -5, and -6; IGF-I; and IGFIR were detected in the retina. In addition, IGF-I mRNA was present in the cornea, IGFBP-1 mRNA was observed in the cornea and iris, and IGFBP-5 and 6 mRNAs were identified in the ciliary body, iris, and cornea. mRNAs for IGFBP-2, -3, and -4 were not found in the eyes. In diabetic rats, reduced levels of IGFBP 6 mRNA were detectable, whereas levels of IGFBP-5 mRNA were increased in the inner and outer retina, rods and cones, iris, cornea, and ciliary body. Other components of the IGF system in the eye were unchanged with diabetes. CONCLUSIONS: In the diabetic (mRen-2)27 rat, IGFBP-6 is downregulated and IGFBP-5 is upregulated by induction of diabetes. Because these IGFBPs may respectively have IGF-enhancing and IGF-inhibitory effects, these findings suggest a possible net IGF-enhancing effect induced by diabetes, providing further evidence for a role of the IGF system in the development of diabetic retinopathy. PMID- 16043843 TI - Oligodendrocyte dysfunction after induction of experimental anterior optic nerve ischemia. AB - PURPOSE: The early response and survival of oligodendrocytes after axonal stroke and their potential contribution to neuronal survival in vivo have not been adequately addressed. The purpose of this study was to investigate the changes occurring in the retina and optic nerve (ON) in anterior ischemic optic neuropathy (AION), using a c-fos transgenic mouse model. METHODS: A new mouse model of AION (rodent AION) was developed to evaluate the in vivo stress response of oligodendrocytes and retinal ganglion cells (RGCs) in a transgenic mouse strain, using the immediate early stress-response gene c-fos, RT-QPCR technology, immunohistochemistry, and electron microscopy. Confocal microscopy was used with cell-specific antibodies to characterize the timing of cells responding to rAION. The TUNEL assay detected cells undergoing apoptosis. Ultrastructural changes were analyzed by electron microscopy. RESULTS: In rAION, oligodendrocytes rapidly respond in vivo to ischemic ON damage, with c-fos activation as an early detectable event. Early evidence of progressive oligodendrocyte stress, is followed by demyelination, wallerian degeneration of the ON, and oligodendrocyte and RGC death far from the primary lesion. CONCLUSIONS: After rAION induction oligodendrocytes, as well as RGCs, undergo progressive stress, with dysfunction and apoptosis. The findings lead to a proposal that progressive retrograde oligodendrocyte stress, away from the primary lesion, is an important factor after ischemic optic neuropathy. Postinduction demyelination must be addressed for effective neuroprotection of ischemic and hypoxic white matter. PMID- 16043844 TI - Identification of a novel splice site mutation of the CSPG2 gene in a Japanese family with Wagner syndrome. AB - PURPOSE: To investigate the genetic basis and clinical variability of Wagner syndrome, a rare, dominantly inherited vitreoretinopathy. METHODS: Clinical examination, linkage analysis, and mutational screening were performed in a large, three-generation, consanguineous Japanese family with Wagner syndrome. The effect of splice site mutation was assessed by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis with lymphoblastoid cell total RNAs generated from affected individuals. RESULTS: Ocular phenotypes of affected members included an empty vitreous with fibrillary condensations, avascular membrane, perivascular sheathing, and progressive chorioretinal dystrophy and were similar to those of the original Wagner syndrome family. All affected eyes examined exhibited pseudoexotropia with ectopic fovea. No systemic manifestations were observed. Genetic linkage confirmed disease segregation with the previously identified WGN1 locus on 5q13-q14. A heterozygous A-->G transversion at the second base of the 3'-acceptor splice site of intron 7 (c.4004-2 A-->G) of the chondroitin sulfate proteoglycan 2 (CSPG2) gene that cosegregated with the disease was identified. Results of RT-PCR analysis indicated that the c.4004-2 A- >G mutation activates a cryptic splice site, located 39 bp downstream from the authentic 3' splice acceptor site. CONCLUSIONS: This linkage study confirmed the genetic homogeneity of the Wagner syndrome. CSPG2 encodes versican, a large chondroitin sulfate proteoglycan, which, in vitreous, binds to hyaluronan and link protein and forms large aggregates that are important for maintaining structural integrity. Although the CSPG2 gene has been excluded as a candidate for causing Wagner syndrome, these data emphasize the necessity of further mutational screening in new families and careful functional characterization. PMID- 16043845 TI - Pigment epithelium-derived factor is a substrate for matrix metalloproteinase type 2 and type 9: implications for downregulation in hypoxia. AB - PURPOSE: Pigment epithelium-derived factor (PEDF), a protein secreted by the retinal pigment epithelium (RPE), acts on retinal survival and angiogenesis. Because hypoxia and VEGF regulate matrix metalloproteinases (MMPs), their effects on PEDF proteolysis were explored. METHODS: Mouse models for retinopathy of prematurity (ROP) were used. Cultured monkey RPE cells were exposed to low oxygen and chemical hypoxia mimetics. PEDF and VEGF mRNA levels in RPE were determined by RT-PCR. MMPs were assessed by zymography, DQ-gelatin degradation solution assays, and MMP immunostaining. PEDF proteolysis was assayed in solution and followed by SDS-PAGE and immunostaining. MMP induction by VEGF was performed in baby hamster kidney (BHK) cells. Retinal R28 cell survival, ex vivo chick embryonic aortic vessel sprouting, and directed in vivo angiogenesis assays were performed. RESULTS: Levels of PEDF in RPE/choroid significantly decreased in the ROP model. Hypoxia decreased PEDF levels in the media conditioned by RPE cells, with no significant change in PEDF mRNA. Conversely, PEDF proteolysis, gelatinolytic activities of approximately 57-kDa and approximately 86-kDa zymogens, and MMP-2 immunoreactivities increased with hypoxia. Addition of VEGF to BHK cells caused a time and dose-related upregulation of approximately 57-kDa zymogens and of DQ-gelatinolytic and PEDF-degrading activity. The PEDF-degrading activity and approximately 57-kDa zymogens in the BHK media shared MMP protease inhibition patterns and MMP-2 immunoreactivities with those in the vitreous. Limited proteolysis with MMP-2 and -9 degraded PEDF in a Ca(+2)-dependent fashion. MMP-mediated proteolysis of PEDF abolished the retinal survival and antiangiogenic activities of the PEDF protein. CONCLUSIONS: Hypoxia and VEGF can downregulate PEDF through proteolytic degradation. PEDF is a novel substrate for MMP-2 and -9. These results reveal a novel posttranslational mechanism for downregulating PEDF, and provide an explanation for hypoxia-provoked increases in VEGF/PEDF ratios, in angiogenesis and/or in neuronal death. PMID- 16043846 TI - Distribution of ocular biometric parameters and refraction in a population-based study of Australian children. AB - PURPOSE: To study the distribution of spherical equivalent refraction and ocular biometric parameters in a young Australian population. METHODS: Noncontact methods were used to examine ocular dimensions and cycloplegic autorefraction in a stratified random cluster sample of year 1 Sydney school students (n = 1765), mean age 6.7 years (range, 5.5-8.4 years). Repeated measures of axial length, anterior chamber depth, and greatest and least corneal radius of curvature (CR1, CR2, respectively) were taken in each eye. Refraction was measured as the spherical equivalent. RESULTS: Mean spherical equivalent refraction in right eyes was +1.26 +/- 0.03 D (SEM; range, -4.88 to +8.58). The distribution was peaked (kurtosis 14.4) and slightly skewed to the right (skewness, 1.7). Prevalence of myopia, defined as spherical equivalent refraction < or = -0.5 D, was 1.43% (95% CI, 0.94-2.18) in the overall population. Axial length, anterior chamber depth, and corneal radii of curvature were normally distributed. The mean axial length in right eyes was 22.61 +/- 0.02 mm (SEM; range, 19.64-25.35). The mean anterior chamber depth was 3.34 +/- 0.01 mm (SEM; range, 2.14-4.06). Mean CR1 was 7.85 +/- 0.01 mm (SEM) and mean CR2 was 7.71 +/- 0.01 mm (SEM). The distribution of axial length/mean corneal radius ratio was peaked (leptokurtic) with a mean of 2.906. Mean axial length was longer, anterior chambers were deeper, and corneas were flatter in the boys. CONCLUSIONS: A peaked (leptokurtic) distribution of spherical equivalent refraction was present in this predominantly hyperopic 6 year-old population. The results also showed that ocular biometric measures were normally distributed, with statistically significant gender differences found in measurements. PMID- 16043847 TI - Converting to SITA-standard from full-threshold visual field testing in the follow-up phase of a clinical trial. AB - PURPOSE: To evaluate the impact of converting from Humphrey 24-2 full-threshold (FT) visual field (VF) testing to SITA-Standard (SS) VF testing during the follow up phase of a clinical trial. METHODS: VF data were obtained from 243 patients in the Collaborative Initial Glaucoma Treatment Study (CIGTS) who had follow-up visits in 2004. FT and SS VF tests were performed in random order on the same day. RESULTS: The average duration of the SS test (6.3 minutes) was shorter (P < 0.0001, paired t-test) than the FT test (11.8 minutes). The mean deviation did not differ between SS and FT testing. A small difference was found in the pattern SD (PSD) (P = 0.02). The mean CIGTS score from the FT test (4.5) was significantly lower (P < 0.0001) than the mean CIGTS score from the SS test (6.0). Although the two tests yielded identical Glaucoma Hemifield Test (GHT) results in 179 patients (76%), 16 patients had a normal GHT result on FT testing and an SS test result that was outside normal limits. Six patients had the reverse finding. The most significant factor associated with an increased (positive) difference between the CIGTS VF score generated from SS and FT testing was conducting the FT test first (P < 0.0001). CONCLUSIONS: Although SS and FT testing yielded very similar mean deviation results, the CIGTS VF score and GHT differed between SS and FT tests. Changing the approach used to measuring a study's primary VF outcome should be accompanied by a critical evaluation of the change's impact. PMID- 16043848 TI - The changing prevalence of myopia in young adults: a 13-year series of population based prevalence surveys. AB - PURPOSE: To determine the changing prevalence of myopia during the years 1990 through 2002 among the 16- to 22-year age group and identify possible risk factors. METHODS: A retrospective study, based on 13 repeated prevalence surveys conducted over a 13-year period. The study subjects were all Israeli nationals belonging to the 16- to 22-year age group from the years 1990 to 2002. Refraction was determined by using subjective visual acuity followed by noncycloplegic autorefraction and subjective validation based on the autorefraction RESULTS: Mild myopia was defined as a refractive error of -0.50 to -3.00 D in at least one eye, moderate myopia as -3.25 to -6.00 D, and high myopia as more than -6.00 D. results. There were 919,929 subjects (382,139 [42%] females and 537,790 [58%] males) included in the study. The overall prevalence of myopia increased from 20.3% in 1990 to 28.3% in 2002. The prevalence of high, moderate, and mild myopia significantly increased in males from 1.7%, 5.7%, and 11.6% in 1990 to 2.05%, 7.2%, and 16.3% in 2002, respectively (P < 0.001). In females, the prevalence of myopia increased from 1.9%, 6.6%, and 13.5% in 1990 to 2.4%, 9.2%, and 20.7% in 2002, respectively (P < 0.001). A correlation between myopia and the number of years of education was observed. Non-Israeli origin was found to be a significant risk factor for myopia. CONCLUSIONS: During the 13 years from 1990 to 2002, the prevalence of myopia significantly increased among the Israeli population. Although there was an association with the level of education, gender, ethnicity, and origin, the prevalence of myopia increased on an annual basis, independent of these factors. PMID- 16043849 TI - The controlled-environment chamber: a new mouse model of dry eye. AB - PURPOSE: To develop a controlled-environment chamber (CEC) for mice and verify the effects of a low-humidity setting on ocular surface signs in normal mice. METHODS: Eight- to 12-week-old BALB/c mice were used in a controlled-environment chamber (CEC) where relative humidity (RH), temperature (T), and airflow (AF) are regulated and monitored. Mice were placed into the CEC and exposed to specific environmentally controlled conditions (RH = 18.5% +/- 5.1%, AF = 15 L/min, T = 21 23 degrees C) for 3, 7, 14, and 28 days. Control mice were kept in a normal environment (RH = 50%-80%, no AF, T = 21-23 degrees C) for the same duration. Aqueous tear production by means of the cotton thread test, corneal fluorescein staining (score, 0-15), and goblet cell density in the superior and inferior conjunctiva were measured by a masked observer. RESULTS: No statistically significant differences between the groups were found at baseline. Decreased tear secretion and increased corneal fluorescein staining were significantly present on day 3, 7, 14, and 28 in animals kept in the CEC. Goblet cell density was significantly decreased in the superior conjunctiva on day 7, and on day 3, 7, and 14 in the inferior conjunctiva in the CEC-kept mice compared with control animals. CONCLUSIONS: This study indicates that exposure of normal mice to a low humidity environment in a CEC can lead to significant alterations in tear secretion, goblet cell density, and acquisition of dry eye-related ocular surface signs. PMID- 16043850 TI - Differential modulation of allergic eye disease by chronic and acute ascaris infection. AB - PURPOSE: To assess alterations in allergic ocular responses to nonparasite antigens in an experimental system in which mice were skewed toward a Th2 cytokine profile by helminth infection. METHODS: Mice were inoculated with Ascaris suum (A. suum) eggs concurrent with ragweed (RW) sensitization (RW/acute) or by repeated inoculation before RW sensitization (RW/chronic). Control subjects were divided into RW, A. suum, and sham-sensitized groups. Animals were RW challenged in the eye and examined for changes in ocular responses, inflammatory cell infiltrates, and in vitro assessment of cytokines after antigen restimulation. In subsequent experiments, CD4(+)/CD25+ T regulatory and CD4(+)/CD25- control T cells were adoptively transferred into mice before ocular challenge. RESULTS: RW sensitization and challenge increased ocular symptoms and eosinophil infiltration into the conjunctiva over PBS control eyes. Acute A. suum infection significantly increased RW-induced clinical symptoms and eosinophil infiltrates in the conjunctiva (P = 0.0001) and resulted in the development of anterior uveitis. In contrast, RW/chronic infection provided protection from allergic responses to RW with significantly fewer eosinophils in the eye and reduced eotaxin levels. Transfer of CD4(+)/CD25+ T cells from RW/chronic mice into RW/acute animals also decreased disease intensity, suggesting that T regulatory cells may contribute to protection from allergic eye disease. CONCLUSIONS: The current studies suggest acute parasitic infections exacerbate allergic symptoms, whereas chronic infections offer protection and provide possible explanations for the role of parasitic infection in susceptibility and resistance to nonparasite allergens. PMID- 16043852 TI - Superior oblique muscle layers in monkeys and humans. AB - PURPOSE: Rectus and the inferior oblique extraocular muscles (EOMs) consist of orbital layers (OLs), inserting on connective tissues, and global layers (GLs), inserting on the sclera. This study was performed to clarify the anatomic relationships of the corresponding layers of the superior oblique (SO) muscle. METHODS: Two whole human and two monkey orbits were serially sectioned en bloc at 10-mum thickness in the coronal plane and stained for collagen with Masson's trichrome and for elastin with van Gieson's stain. The SO muscles of one human and one monkey were sectioned longitudinally. The structure of the SO muscle was examined by light microscopy, and muscle fibers in the OL and GL of selected sections were counted. RESULTS: The deep SO muscle consisted of a central GL contiguous with the tendon, surrounded coaxially by a peripheral OL inserting on the SO sheath posterior to the trochlea. The maximum number of SO fibers was 14,400 to 19,200 in the human and 7,000 to 7,400 in the monkey. In the monkey, approximately 60% of total fibers were in the GL, and 40% in the OL. The SO sheath was in mechanical continuity with the superior rectus pulley. CONCLUSIONS: The primate SO has a substantial OL configured to contribute to positioning the superior rectus pulley in the coronal plane. Whereas the direction of application of the SO's GL force is determined by the rigid trochlea, the SO's OL influences the direction of application of rectus EOM forces. This insight extends the concept of active control of pulley positions to include a contribution from the SO muscle. PMID- 16043851 TI - Nitric oxide and cGMP mediate alpha1D-adrenergic receptor-Stimulated protein secretion and p42/p44 MAPK activation in rat lacrimal gland. AB - PURPOSE: To determine whether alpha(1)-adrenergic receptors use the nitric oxide (NO)/cGMP pathway to stimulate protein secretion by rat lacrimal gland. METHODS: Identification and cellular location of endothelial nitric oxide synthase (eNOS) and neuronal nitric oxide synthase (nNOS) were determined by Western blot and immunofluorescence techniques, respectively. Rat lacrimal gland acini were isolated by collagenase digestion, and protein secretion stimulated by phenylephrine, an alpha(1)-adrenergic agonist, was measured with a fluorescence assay system. Acini were preincubated with inhibitors for 20 minutes before addition of phenylephrine (10(-4) M). NO and cGMP were measured in response to phenylephrine stimulation. Activation of p42/p44 MAPK was determined by Western blot analysis with an antibody against phosphorylated (active) p42/p44 MAPK. RESULTS: eNOS and nNOS were both present in lacrimal gland. eNOS appeared to be localized with caveolae, whereas nNOS was present in the nerves surrounding the acini. Inhibition of eNOS with N(G)-nitro-l-arginine methyl ester (l-NAME; 10(-6) M) completely inhibited phenylephrine-stimulated protein secretion, whereas the inactive isomer d-NAME and inhibition of nNOS with S-methyl-l-thiocitrulline did not. Phenylephrine increased NO production in a time- and concentration-dependent manner, but the increase was abolished by the alpha(1D)-adrenergic receptor inhibitor BMY-7378. Inhibition of guanylate cyclase with oxadiazoloquinoxalin (ODQ) also inhibited phenylephrine-induced protein secretion, whereas phenylephrine caused a 2.2-fold increase in cGMP. In addition, preincubation with l-NAME and ODQ inhibited phenylephrine-stimulated p42/p44 MAPK activation. CONCLUSIONS: alpha(1D)-Adrenergic agonists stimulate eNOS to produce NO, leading to production of cGMP by guanylate cyclase, to transduce the extracellular signal through the cell and stimulate protein secretion in rat lacrimal gland. PMID- 16043853 TI - Kinematics of vertical saccades during the yaw vestibulo-ocular reflex in humans. AB - PURPOSE: Listing's law (LL) constrains the rotational axes of saccades and pursuit eye movements to Listing's plane (LP). In the velocity domain, LL is ordinarily equivalent to a tilt in the ocular velocity axis equal to half the change in eye position, giving a tilt angle ratio (TAR) of 0.5. This study was undertaken to investigate vertical saccade behavior after the yaw vestibulo ocular reflex (VOR) had driven eye torsion out of LP, an initial condition causing the position and velocity domain formulations of LL to differ. METHODS: Binocular eye and head motions were recorded with magnetic search coils in eight humans. With the head immobile, LP was determined for each eye, and mean TAR was 0.50 +/- 0.07 (mean +/- SD) for horizontal and 0.45 +/- 0.11 for vertical saccades. The VOR was evoked by transient, whole-body yaw at 2800 deg/s2 peak acceleration, capable of evoking large, uninterrupted VOR slow phases. Before rotation, subjects viewed a target at eye level, 20 degrees up, or 20 degrees down. In two thirds of the trials, the target moved upward or downward at systematically varying times, triggering a vertical saccade during the horizontal VOR slow phase. RESULTS: Because the head rotation axis was generally misaligned with LP, the eye averaged 3.6 degrees out of LP at vertical saccade onset. During the saccade, eye position continued to depart LP by an average 0.8 degrees. The horizontal TAR at saccade onset was 0.29 +/- 0.07. At peak saccade velocity 35 +/ 3 ms later, the vertical TAR was 0.45 +/- 0.07, statistically similar to that of head fixed saccades. Saccades did not return to LP. CONCLUSIONS: Although they did not observe the position domain formulation of LL, vertical saccades, during the VOR, observed the half-angle velocity domain formulation of LL. PMID- 16043854 TI - Binocular summation of detection and resolution thresholds in the central visual field using parallel-line targets. AB - PURPOSE: To investigate the effect of thresholds on binocular summation for detection and resolution of parallel-line targets of different widths. METHODS: The automatic perimeter Octopus 201 (Haag-Streit International, Koniz, Switzerland) combined with a modified haploscope device and targets consisting of parallel lines that were 10' (10' target), 2.5' (2.5' target), or 1.43' (1.43' target) of visual angle in width and apart were used to measure the thresholds for monocular-binocular detection and resolution of the targets under the same binocular fusion stimulation conditions in seven young adults with normal vision. The custom program in the automatic perimeter was used to test 27 points. Seventeen of the 27 points were located in the central 6 degrees of visual field and 10 on the horizontal meridian subtending visual angles of 8 degrees , 10 degrees , 12 degrees , 16 degrees , and 20 degrees . RESULTS: The resolution threshold was significantly higher than the detection threshold for the 2.5' and 1.43' targets (P < 0.01 by Bonferroni/Dunn test). Furthermore, as the target width decreased to 2.5' or 1.43', the binocular summation ratio for the resolution threshold increased significantly over that for the detection threshold, with increasing distance from the fovea (P < 0.01 by Wilcoxon signed ranks test). CONCLUSIONS: Binocular summation for detection and resolution thresholds varies as a function of the width of a parallel-line target. The difference between binocular summation ratios for detection and resolution thresholds increases with decrease in target width and increase in eccentricity from the fovea. Binocular interaction plays an important role in the task of recognizing a high-resolution target in the parafoveal area. PMID- 16043855 TI - Clinical features and course of patients with glaucoma with the E50K mutation in the optineurin gene. AB - PURPOSE: To investigate the clinical features of subjects with glaucoma with the E50K mutation in the optineurin (OPTN) gene and to compare the onset, severity, and clinical course of these patients with a control group of subjects with glaucoma without this mutation. METHODS: The phenotype of well-characterized subjects from Moorfields Eye Hospital, London, who had been identified as carrying the OPTN E50K mutation was examined. A wide range of structural, psychophysical, and demographic factors were then compared with those in a control group of subjects with glaucoma without this mutation. RESULTS: Eleven subjects with glaucoma with the E50K mutation (nine in two families and two sporadic cases) were studied. All 11 subjects had normal tension glaucoma (NTG), with presenting and highest IOP of 15.3 +/- 3.0 and 16.5 +/- 2.5 mm Hg (+/-SD) on diurnal testing. Compared with 87 NTG control subjects who did not have this mutation, subjects with E50K presented at a younger age (40.8 +/- 15 years, P = 0.0001) and had more advanced optic disc cupping (mean cup-disc ratio +/- SD 0.86 +/- 0.1, P = 0.001) and smaller neuroretinal rim area (+/-SD; 0.5 +/- 0.28 mm2, P = 0.001) at diagnosis. The rate of filtration surgery performed for progressive visual field loss in those with and without the E50K mutation was 72.7% and 25.3%, respectively (P = 0.003), and all subjects with E50K were found to have progressing visual fields. In addition, seven E50K mutation-carrying individuals in two families (age range, 23-58 years) presented with normal optic discs and visual fields and, as yet, no signs of glaucoma. CONCLUSIONS: In this study, subjects with glaucoma who had the OPTN E50K mutation were found to have NTG that appeared to be more severe than that in a control group of subjects with NTG without this mutation. The findings emphasize the importance of early detection and treatment of glaucoma in such individuals, to minimize visual loss. PMID- 16043856 TI - Laser scanning tomography of the optic nerve head in a normal elderly population: the Bridlington eye assessment project. AB - PURPOSE: To assess optic nerve head topographic parameters using the Heidelberg Retina Tomograph (HRT) II (Heidelberg Engineering GmbH, Dossenheim, Germany) in a normal elderly population. METHODS: Optic nerve head analysis of 918 eyes of 459 normal elderly patients was performed. All patients were consecutive in a cohort screened for eye disease. Normal subjects were defined with a normal visual field on automated suprathreshold screening, intraocular pressure less than 22 mmHg, and minimum corrected visual acuity of 6/12. All optic discs were contoured by two investigators and the mean parameters analyzed. The effects of age, sex, and disc size were assessed. RESULTS: Subjects' (262 women and 197 men) mean age was 72.6 +/- 5.1 (SD) years (range, 65.5-89.3). Mean +/- SD global disc area, cup/disc area ratio, and neuroretinal rim area were 1.98 +/- 0.36 mm2, 0.22 +/- 0.14, and 1.52 +/- 0.31 mm2, respectively. Disc area did not differ significantly based on eye side or sex. The women were found to have a significantly larger rim volume, mean retinal nerve fiber layer (RNFL) thickness, and cross-sectional area than the men and tended to have smaller cup areas/volumes and cup/disc area ratios. Most tomography parameters were found to be significantly influenced by disc size. CONCLUSIONS: To the authors' knowledge, this is the first large study of optic nerve head parameters in the elderly normal population using the HRT II. This age range is particularly relevant to glaucoma detection and pertinent to discriminant analyses separating normal subjects from glaucoma in screening for the disease. Given the systematic differences between the parameters in men and women, reference ranges should be quoted by sex. PMID- 16043857 TI - Hypophosphorylation of aqueous humor sCD44 and primary open-angle glaucoma. AB - PURPOSE: The ectodomain of CD44, the principal receptor for hyaluronic acid (HA), is shed as a 32-kDa fragment-soluble CD44 (sCD44)-which is cytotoxic to trabecular meshwork (TM) cells and retinal ganglion cells (RGCs) in culture. The purpose of this study was to characterize sCD44 further by determining the phosphorylation of aqueous humor sCD44 in normal and primary open-angle glaucoma (POAG). METHODS: Aqueous humor samples of patients were subjected to CD44 enzyme linked immunosorbent assay (ELISA) and two-dimensional (2-D) polyacrylamide gel electrophoresis, followed by Western blot analysis with anti-CD44, anti serine/threonine, and anti-tyrosine phosphospecific antibodies, to determine sCD44 concentration, isoelectric point (pI), and phosphorylation, respectively. The bioactivity of hypophosphorylated sCD44 was tested in cell culture and HA affinity columns. RESULTS: Two-dimensional Western blot analysis revealed that the representative pI of the 32-kDa sCD44 was 6.96 +/- 0.07 in POAG versus 6.38 +/- 0.08 in normal (P < 0.0004). Enzymatic dephosphorylation of sCD44 resulted in a basic shift in the pI. The normal aqueous humor sCD44 was positive for serine threonine phosphorylation; however, POAG sCD44 was hypophosphorylated. Hypophosphorylated sCD44 was more toxic to TM and RGC cells than standard sCD44, and hypophosphorylated sCD44 had decreased affinity to HA, particularly with increased pressure. CONCLUSIONS: POAG aqueous is characterized by posttranslational change in the pI of sCD44 and hypophosphorylation, which clearly distinguished POAG from normal aqueous humor. The high toxicity and low HA-binding affinity of hypophosphorylated sCD44 may represent specific pathophysiologic features of the POAG disease process. PMID- 16043858 TI - Analysis of GDx-VCC polarimetry data by Wavelet-Fourier analysis across glaucoma stages. AB - PURPOSE: The purpose of this study was to apply shape-based analysis techniques of retinal nerve fiber layer (RNFL) thickness to GDx-VCC (variable corneal and lens compensator; Laser Diagnostic Technologies, Inc., San Diego, CA) polarimetry data and to evaluate the techniques' ability to detect glaucoma in its earliest stages. Wavelet-based (wavelet-Fourier analysis [WFA]), Fourier-based (fast Fourier analysis [FFA]), and several previous variations of shape-based analysis were considered, as well as the standard metric nerve fiber indicator (NFI), and all were compared as a function of disease stage. METHODS: GDx-VCC scans of one eye of each of 67 patients with glaucoma and each of 67 healthy age-matched subjects provided RNFL thickness estimates at a fixed distance from the optic disc. Severity of disease was graded according to the Glaucoma Staging System and also by mean deviation (MD) from standard automated perimetry. WFA, FFA, and NFI procedures were performed including the following variations: use of signed or unsigned phase, inclusion of interocular or intraocular asymmetry of analysis parameters, and combination of features by principle components analysis or Wilks lambda. Independent samples (k-fold variation) were used for training and testing. Sensitivity, specificity, and receiver operating characteristic (ROC) area were obtained. RESULTS: Classification performance of WFA (ROC = 0.978) was significantly better than FFA (ROC = 0.938) and NFI (ROC = 0.900). This difference was largest for the earliest stages of glaucoma. Shape-based analysis methods performed better than NFI overall. Adding between-eye asymmetry measures helped FFA but not WFA. CONCLUSIONS: Shape-based analysis, and WFA in particular, makes an important improvement in detecting earliest glaucoma with polarimetry. PMID- 16043859 TI - Primary trabecular meshwork cells incubated in human aqueous humor differ from cells incubated in serum supplements. AB - PURPOSE: To determine whether aqueous humor, the in vivo source of nutrients for trabecular meshwork cells, alters cellular and molecular characteristics in primary trabecular monolayer cell cultures when compared with standard culture conditions. METHODS: Human primary trabecular meshwork cell cultures were grown in DMEM supplemented with 50% human aqueous humor (DMEM-AH), heat-denatured DMEM AH, 10% fetal bovine serum (DMEM-FBS, the standard culture supplement), or heat denatured DMEM-FBS. Confluent trabecular cells were assayed for cell propagation and morphology for 21 days. Protein expression profiles of trabecular cell lysates were analyzed by two-dimensional polyacrylamide gel electrophoresis. Western blot analysis was used to determine the protein expression of myocilin and TIMP-1 in conditioned media collected from trabecular cells at 5, 10, 15, and 21 days. Myocilin expression was also analyzed by Western immunoblots after addition of dexamethasone (10(-7) M) or ascorbic acid (29 mg/dL). RESULTS: Trabecular cells supplemented with DMEM-AH for 21 days showed decreased cell proliferation when compared with DMEM-FBS (11% vs. 141%). Cellular morphology was also altered: Trabecular cells incubated in DMEM-AH showed larger-, broader-, and flatter-appearing cells than did the more spindle-shaped cells grown in DMEM-FBS. Protein profiles of trabecular cell lysates isolated from cells incubated in DMEM AH differed from those incubated in DMEM-FBS. In DMEM-AH-conditioned medium, myocilin expression was increased and TIMP-1 expression was decreased at day 21. Induction of myocilin by dexamethasone was observed in conditioned medium isolated from cells treated with DMEM-FBS (442%), but only a 10% increase in myocilin was observed beyond the normal induction in DMEM-AH. Daily administration of ascorbic acid to DMEM-AH failed to increase myocilin expression beyond that obtained with DMEM-AH. CONCLUSIONS: Addition of human aqueous humor rather than FBS to trabecular monolayer cell cultures triggers significant changes in cellular and molecular characteristics. The protein component of aqueous humor is responsible for these changes. Aqueous humor supplementation may maintain cultured trabecular cells in a more physiologic state. PMID- 16043860 TI - Changes in gene expression by trabecular meshwork cells in response to mechanical stretching. AB - PURPOSE: Trabecular meshwork (TM) cells appear to sense changes in intraocular pressure (IOP) as mechanical stretching. In response, they make homeostatic corrections in the aqueous humor outflow resistance, partially by increasing extracellular matrix (ECM) turnover initiated by the matrix metalloproteinases. To understand this homeostatic adjustment process further, studies were conducted to evaluate changes in TM gene expression that occur in response to mechanical stretching. METHODS: Porcine TM cells were subjected to sustained mechanical stretching, and RNA was isolated after 12, 24, or 48 hours. Changes in gene expression were evaluated with microarrays containing approximately 8000 cDNAs. Select mRNA changes were then compared by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Western immunoblots were used to determine whether some of these changes were associated with changes in protein levels. RESULTS: On the microarrays, 126 genes were significantly upregulated, and 29 genes were significantly downregulated at one or more time points, according to very conservative statistical and biological criteria. Of the genes that changed, several ECM regulatory genes, cytoskeletal-regulatory genes, signal-transduction genes, and stress-response genes were notable. These included several proteoglycans and matricellular ECM proteins composed of common repetitive binding domains. The results of analysis of mRNA changes in more than 20 selected genes by qRT-PCR supported the findings in the microarray analysis. Western immunoblots of several proteins demonstrated protein level changes associated with changes in the level of mRNA. CONCLUSIONS: The expression of a variety of TM genes is significantly affected by mechanical stretching. These include several ECM proteins that contain multiple binding sites and may serve organizational roles in the TM. Several proteins that could contribute to the homeostatic modification of aqueous humor outflow resistance are also upregulated or downregulated. PMID- 16043861 TI - Molecular characterization of the cystine/glutamate exchanger and the excitatory amino acid transporters in the rat lens. AB - PURPOSE: To determine whether the cyst(e)ine/glutamate exchanger (XC-) and the excitatory amino acid transporters (EAAT1 to -5) are expressed in the rat lens. METHODS: A combination of molecular-based and immunocytochemical strategies was used to screen for the presence of the light-chain subunit of XC- (xCT) and the five known EAAT isoforms in the rat lens. An initial molecular profiling of xCT and EAAT1 to -5 expression was achieved by reverse transcription-polymerase chain reaction (RT-PCR). The presence of transporter proteins was verified by Western blot analysis and immunocytochemistry. RESULTS: Transcripts for xCT and EAAT1 to 5 were detected by RT-PCR in lens fiber cells. Western blot analysis confirmed the expression of xCT and all five EAAT isoforms at the protein level. Immunocytochemistry revealed xCT expression to be present throughout the lens. Notably, changes in the subcellular distribution of xCT were shown to occur as a function of fiber cell differentiation. In the outer cortex, xCT labeling was predominantly cytoplasmic but progressively became more membranous with distance into the lens, due to xCT insertion into the broad sides of fiber cells. In the core, xCT labeling was localized around the entire membrane of inner fiber cells suggesting a redistribution of the exchanger. In contrast, EAAT expression was restricted to the outer cortex of the lens, with EAAT4/5 shown to be the predominant isoforms in cortical fiber cells. Western blot analysis of crude fiber membranes dissected from the outer cortex, inner cortex, and core region of the lens confirmed the presence of xCT in all three of these regions and demonstrated that EAATs were absent from the core region. CONCLUSIONS: The molecular identification and localization of xCT and EAAT1 to -5 in the lens raises the possibility that in the outer cortex XC- and EAAT4/5 may work together to accumulate cysteine for GSH synthesis. The presence of xCT and the absence of the EAATs in the center of the lens suggest that XC- could operate with an alternative glutamate uptake pathway to accumulate cysteine where it can potentially act as a low-molecular-mass antioxidant. PMID- 16043862 TI - Monocarboxylate transporter expression remains unchanged during the development of diabetic retinal neuropathy in the rat. AB - PURPOSE: To determine the effect of diabetes on monocarboxylate transporter (MCT) expression in the rat retina. METHODS: Diabetes was induced in Wistar rats by a single intraperitoneal injection of streptozotocin (62.5 mg/kg). Rats were killed after 10 weeks, and the retinal levels of PKCalpha, bFGF, glial fibrillary acidic protein (GFAP), caspase-3, and MCT1, -2, and -4 were assessed with immunoblot analysis and RT-PCR. Hippocampal samples were used as a comparison. In other animals, the retinas were processed histologically and sections stained for the breakdown of DNA (TUNEL procedure) and for the distribution of MCT1, -2, and -4. RESULTS: Diabetic rats exhibited cataract formation, elevated blood glucose levels, and polydipsia. Retinal levels and distribution of MCT1, 2, and -4 were similar in the diabetic and age-matched control groups; however, the early retinopathy markers bFGF and PKCalpha were significantly elevated in the diabetic retinas but not in the hippocampal samples. ERG recordings showed decreased oscillatory potentials in the diabetic group, and TUNEL staining was most evident in the photoreceptor layer. Activated caspase-3 levels were elevated in the diabetic retina. CONCLUSIONS: The expression of retinal MCT1, -2, and -4 is unaffected after 10 weeks of diabetes in rats. It appears unlikely that diabetic retinal neuropathy is a result of a hyperglycemic induction of altered MCT expression. PMID- 16043863 TI - Eye-movement training for reading in patients with age-related macular degeneration. AB - PURPOSE: To determine whether training oculomotor control, without direct practice in reading sentences, could increase reading speed in patients with age related macular degeneration (AMD). METHODS: Sixteen patients with AMD participated in the study (age range, 65-87 years; mean, 77). The training program consisted of a series of exercises that were designed to allow the patients to practice eye movements. At the beginning of training, the subjects practiced small horizontal saccades in response to cognitively easy stimuli (e.g., dots). The training then progressed to practicing larger eye movements and then to practicing saccades with single letters, pairs of letters, and three letter words. Reading of sentences was practiced in only one exercise, during the last session of the 8-week training. RESULTS: The difference between average reading speeds before and after training was 24.7 wpm (difference between medians, 17.9 wpm). The increase in speed was statistically significant (Wilcoxon signed rank test = 124.0, P < 0.001). There was no significant relationship between change in maximum reading speed and ETDRS (Early Treatment Diabetic Retinopathy Study) acuity (r = -0.14, P = 0.76) or between change in maximum reading speed and age (r = 0.25, P = 0.45). CONCLUSIONS: The results indicate that a training curriculum that concentrates on eye-movement control can increase reading speed in patients with AMD. This finding is especially interesting, because the training involved little direct practice in reading sentences but instead concentrated on having subjects practice control of eye positions and eye movements. PMID- 16043864 TI - Retinoid X receptor (gamma) is necessary to establish the S-opsin gradient in cone photoreceptors of the developing mouse retina. AB - PURPOSE: The retinoid X receptors (RXRs) are members of the family of ligand dependent nuclear hormone receptors. One of these genes, RXRgamma, is expressed in highly restricted regions of the developing central nervous system (CNS), including the retina. Although previous studies have localized RXRgamma to developing cone photoreceptors in several species, its function in these cells is unknown. A prior study showed that thyroid hormone receptor beta2 (TRbeta2) is necessary to establish proper cone patterning in mice by activating medium wavelength (M) cone opsin and suppressing short-wavelength (S) cone opsin. Thyroid hormone receptors often regulate gene transcription as heterodimeric complexes with RXRs. METHODS: To determine whether RXRgamma cooperates with TRbeta2 to regulate cone opsin patterning, the developmental expression of RXRgamma was examined, and cone opsin expression in RXRgamma-null mice was analyzed. RESULTS: RXRgamma was expressed in postmitotic cones and was transiently downregulated at the time of S-opsin onset in both mouse and human cones. RXRgamma-null mice expressed S-opsin in all cones, similar to the TRbeta2 null mice. Unlike TRbeta2-null mice, which did not express M-opsin, RXRgamma-null mice had a normal pattern of M-opsin expression. CONCLUSIONS: RXRgamma is essential (along with TRbeta2) for suppressing S-opsin in all immature cones and in dorsal cones in the mature retina, but it is not necessary for M-opsin regulation. These results demonstrate a critical role for RXRs in regulating cell differentiation in the CNS and highlight a remarkable conservation of opsin regulation from Drosophila to mammals. PMID- 16043865 TI - Targeted disruption of FSCN2 gene induces retinopathy in mice. AB - PURPOSE: To investigate the morphology and function of photoreceptors in mice with mutation of the FSCN2 gene. METHODS: A mouse line was generated carrying the 208delG mutation (point mutation, or p-type) and another with replacement of exon 1 by the cDNA of a green fluorescent protein (GFP knock-in, or g-type). The expression of retinal mRNA was determined by reverse transcription (RT) polymerase chain reaction (PCR) and in situ hybridization performed on retinal sections. Morphologic analyses of the retinas were performed by light microscopy (LM) and transmission electron microscopy (TEM) and functional analyses by electroretinogram (ERG). RESULTS: mRNA of FSCN2 was not detected in the retinal mRNA extracted from FSCN2p/p and FSCN2g/g mice. Both FSCN2(+/p) and FSCN2(+/g) mice had progressive photoreceptor degeneration with increasing age detected by LM and structural abnormalities of the outer segment (OS) detected by TEM. Both FSCN2(+/p) and FSCN2(+/g) mice had depressed rod and cone ERGs that worsened with increasing age. CONCLUSIONS: These results indicate that haploinsufficiency of the FSCN2 gene may hamper maintenance and/or elongation of the OS disks and result in photoreceptor degeneration, as in human autosomal dominant retinitis pigmentosa. PMID- 16043866 TI - The RAGE axis in early diabetic retinopathy. AB - PURPOSE: The receptor for advanced glycation end products (AGEs) has been implicated in the pathogenesis of diabetic complications. This study was conducted to characterize the role of the RAGE axis in a murine model of nonproliferative diabetic retinopathy (NPDR). METHODS: The retinas of hyperglycemic, hyperlipidemic (HGHL, apolipoprotein E(-/-) db/db) mice were examined for the development of early retinal vascular lesions of NPDR and compared to littermates at 6 months of age. Neural function was assessed with electroretinography. Immunohistochemistry, real-time RT-PCR, autofluorescence, and ELISA studies were used to localize and quantify the AGE/RAGE axis. Soluble RAGE, a competitor of cellular RAGE for its ligands, was administered to assess the impact of RAGE blockade. RESULTS: Early inner retinal neuronal dysfunction, manifested by prolonged latencies of the oscillatory potentials and b-wave, was detected in hyperglycemic mice. HGHL mice exhibited accelerated development of acellular capillaries and pericyte ghosts compared with littermate control animals. AGEs were localized primarily to the vitreous cavity and internal limiting membrane (ILM) of the retina, where they were intimately associated with the footplates of RAGE-expressing Muller cells. AGE accumulation measured by ELISA was increased within the retinal extracellular matrix of hyperglycemic mice. AGE fluorescence and upregulation of RAGE transcripts was highest in the retinas of HGHL mice, and attenuation of the RAGE axis with soluble RAGE ameliorated neuronal dysfunction and reduced the development of capillary lesions in these mice. CONCLUSIONS: In early diabetic retinopathy, the RAGE axis, comprising the cellular receptor and its AGE ligands, is amplified within the retina and is accentuated along the vitreoretinal interface. Antagonism of the RAGE axis in NPDR reduces neurovascular perturbations, providing an important therapeutic target for intervention. PMID- 16043867 TI - Suppression of ocular inflammation in endotoxin-induced uveitis by blocking the angiotensin II type 1 receptor. AB - PURPOSE: To examine whether the angiotensin II type 1 receptor (AT1-R) signaling plays a role in ocular inflammation in endotoxin-induced uveitis (EIU). METHODS: EIU was induced in C57BL/6 mice by a single intraperitoneal injection of 150 mug lipopolysaccharide (LPS). Tissue localization, mRNA expression, and protein levels of AT1-R in murine retinas were examined by immunohistochemistry, RT-PCR, and Western blot analyses, respectively. Telmisartan, an AT1-R antagonist widely used as an antihypertensive agent, was administered intraperitoneally at a dose of 10 mg/kg daily for 5 days until the injection of LPS. Twenty-four hours after administration, leukocyte adhesion to the retinal vasculature was evaluated with a concanavalin A lectin perfusion-labeling technique. Retinal mRNA and protein levels of intercellular adhesion molecule (ICAM)-1 were examined by RT-PCR and ELISA, respectively. Protein concentration and inflammatory cells in the aqueous humor were also measured. RESULTS: Retinal vessels were positive for AT1-R. In mice with EIU, retinal AT1-R mRNA and protein levels were significantly increased when compared to the normal control. EIU animals also showed significant increases in the number of inflammatory cells infiltrating the anterior chamber and adhering to the retinal vessels and in retinal ICAM-1 levels. Administration of telmisartan to EIU mice resulted in significant suppression of retinal ICAM-1 expression and leukocyte adhesion and infiltration compared with vehicle treatment. Protein concentration in the aqueous humor of telmisartan-treated EIU mice tended to be lower than that of vehicle-treated EIU mice, but the difference was not statistically significant. CONCLUSIONS: AT1-R signaling blockade inhibited retinal ICAM-1 upregulation and leukocyte adhesion and infiltration in the EIU model. These results suggest the potential use of an AT1-R antagonist as a therapeutic agent to reduce ocular inflammation. PMID- 16043868 TI - A system for inducible gene expression in retinal ganglion cells. AB - PURPOSE: To develop a system for inducible gene expression in retinal ganglion cells, Thy1 and ckit promoters were used to direct expression of a second generation reverse tetracycline transactivator (rtTA2S-M2). METHODS: Transgenic mice were generated that harbor rtTA2S-M2 under the control of either the Thy1 or ckit promoter. These animals were crossed with mice transgenic for the LacZ gene downstream of a cassette of tet operator (TRE) binding sites. Induction of the LacZ reporter gene in vivo after either oral or subcutaneous doxycycline administration and in vitro in cultured retinal cells was assessed. To examine induction of a secreted protein, expression of pigment epithelium-derived factor (PEDF) in mice harboring Thy1-rtTA and TRE-PEDF constructs was quantified. RESULTS: Five of seven Thy1-rtTA lines showed induction with subcutaneous doxycycline: maximum induction in one line (Thy1-C), moderate in one line (Thy1 F), and minimal in three lines. There was no detectable retinal LacZ expression in the ckit-rtTA lines, despite expression of the ckit-rtTA transgene at the RNA level. In Thy1-rtTA lines, LacZ reporter expression as measured by X-gal staining was evenly dispersed throughout all quadrants of the retina, present in a subpopulation of retinal ganglion cell (RGC) bodies, RGC axons projecting through the retina and optic nerve, and some cells in the inner nuclear layer. Immunostaining for beta-galactosidase demonstrated more uniform expression in RGCs and cells of the inner aspect of the inner nuclear layer, which, by double staining with anti-beta-galactosidase and anti-calretinin antibodies, were consistent with amacrine cells. More than 95% of Thy-1 antigen-positive cells in the retina expressed the induced transgene. Subcutaneous doxycycline resulted in a more robust induction of LacZ than did oral administration. In vitro, the number of cells induced in culture increased in a dose-dependent manner, with maximum expression at 10 microg/mL at a level 3.4-fold over background. Thy1 rtTA/TRE-PEDF mice treated with doxycycline had 1000-fold induction in their retinal PEDF expression in comparison with nontransgenic mice and 600-fold induction over noninduced Thy1-rtTA/TRE-PEDF mice. CONCLUSIONS: A transgenic system for inducible RGC expression has been developed that demonstrates minimal leakiness and significant induction with doxycycline. This system will be useful for several applications. PMID- 16043869 TI - Autofluorescence characteristics of normal foveas and reconstruction of foveal autofluorescence from limited data subsets. AB - PURPOSE: To develop mathematical and geometric models of the nonuniform autofluorescence (AF) patterns of foveas of normal subjects and to reconstruct these models from limited subsets of data. METHODS: Confocal scanning laser ophthalmoscope (cSLO) AF fundus images of normal maculae were obtained from both eyes of 10 middle-aged subjects. They were filtered and contrast enhanced, to obtain elliptical isobars of equal gray levels (GLs) and determine the isobars' resolutions, eccentricities, and angles of orientation. The original image data were fit with a mathematical model of elliptic quadratic polynomials in two equal zones: the center and the remaining annulus. RESULTS: The AF images segmented into nested concentric GL isobars with GLs that increased radially from the least fluorescent center. The mean isobar resolution was 31 +/- 7 mum. The geometric eccentricity of the ellipses increased from 0.42 +/- 0.12 centrally to 0.52 +/- 0.14 peripherally (P = 0.0005), with mean axes of orientation peripherally 97.12 +/- 15.46 degrees . The model fits to the complete image data had mean absolute normalized errors ranging from 3.6% +/- 3.7% to 7.3% +/- 7.1%. The model fits to small subsets (1% to 2% of total image data) had mean absolute errors ranging from 3.7% +/- 3.8% to 7.3% +/- 7.2%. CONCLUSIONS: Normal AF fundus images show finely resolved, concentric, elliptical foveal patterns consistent with the anatomic distribution of fluorescent lipofuscin, light-attenuating macular pigment (MP), cone photopigment, and retinal pigment epithelial (RPE) pigment in the fovea. A two-zone, elliptic, quadratic polynomial model can accurately model foveal data. This model may be useful for image analysis and for automated segmentation of pathology. PMID- 16043870 TI - Prolactins are natural inhibitors of angiogenesis in the retina. AB - PURPOSE: Disruption of the normally antiangiogenic environment of the retina leads to aberrant angiogenesis, the major cause of vision loss throughout the world. Prolactin (PRL), the hormone originally associated with milk production, can be proteolytically processed to 16K-PRL, a 16 kDa N-terminal PRL fragment with potent antiangiogenic and vasoconstrictive actions. This study was conducted to determine whether 16K-PRL is found naturally in the retina and plays a role in angiogenesis and vasodilation. METHODS: Expression of PRL mRNA in rat retina was determined by RT-PCR and in situ hybridization. Western blot was used to examine the expression of PRL and derived fragments in retinal homogenates. The role of PRL and 16K-PRL in the retina was studied by intravitreal injection of either antibodies against PRL or small interfering RNAs (siRNA), to suppress expression of retinal PRL mRNA. RESULTS: Rat retina expressed PRL mRNA in the outer nuclear, outer plexiform, inner nuclear, and ganglion cell layers. Both full-length PRL and N-terminal 16K-PRL were detected in retinal homogenates by polyclonal and monoclonal antibodies. The intravitreal injection of antibodies able to neutralize the actions of 16K-PRL increased the number of retinal blood vessels and capillary area by threefold. Furthermore, siRNA-mediated inhibition of PRL mRNA increased retinal neovascularization threefold and resulted in a significant increase in vasodilation. CONCLUSIONS: These results demonstrate that PRL is synthesized and cleaved to antiangiogenic 16K-PRL by retinal tissue and that these molecules play a key role in preventing angiogenesis in the healthy retina. PMID- 16043871 TI - Drug intervention can correct subnormal retinal oxygenation response in experimental diabetic retinopathy. AB - PURPOSE: Early subnormal retinal oxygenation response to a hyperoxic provocation (DeltaPo2) is strongly associated with subsequent experimental diabetic retinopathy and can be reversed by drug treatments started with the induction of diabetes. It is not yet known whether drug treatment can reverse an established subnormal DeltaPo2. METHODS: Retinal DeltaPo2 was measured in two separate experimental paradigms in streptozotocin-induced diabetic rats. In a prevention study, measurements were performed in untreated diabetic rats, 3 months after the initiation of hyperglycemia (D3mo), in age-matched nondiabetic rats (C3mo), and in diabetic rats treated orally for 3 months with celecoxib, a cyclooxygenase (COX)-2-selective inhibitor, (D3mo+COX2i). In an intervention study, measurements were performed in untreated diabetic rats 4 months after the initiation of diabetes (D4mo), in age-matched nondiabetic rats (C4mo), and in diabetic rats that were untreated for 3 months and then were orally treated for an additional month with either celecoxib (D4mo+COX2i) or l-N (6)-(1-iminoethyl)lysine 5 tetrazole amide, a prodrug of an inhibitor of inducible nitric oxide synthase (iNOS, D4mo+ iNOSi). RESULTS: In the prevention arm, subnormal (P < 0.05) retinal DeltaPo2 was found in the D3mo group, but not in the D3mo+COX2i group (P > 0.05). In a previous study, it was reported that retinal DeltaPo2 also corrected in a D3mo+iNOSi group. In the intervention arm, retinal DeltaPo2 levels in the D4mo and D4mo+iNOSi, but not the D4mo+COX2i, groups were (P < 0.05) subnormal. CONCLUSIONS: These results demonstrate, for the first time, that drug treatment can reverse an established subnormal DeltaPo2. Furthermore, this effect could not be predicted by a drugs' ability to prevent the development of subnormal DeltaPo2. PMID- 16043872 TI - Retinal contraction and metamorphopsia scores in eyes with idiopathic epiretinal membrane. AB - PURPOSE: Using M-CHARTS (Inami Co., Tokyo, Japan), which were developed by the authors to measure metamorphopsia, and image-analysis software, which was developed to quantify retinal contraction, the authors investigated the relationship between the degree of retinal contraction and the degree of metamorphopsia in eyes with idiopathic epiretinal membrane (ERM). METHODS: This study was conducted in 29 eyes with ERM (29 patients, 20 women; mean age, 62.1 +/ 8.6 years) observed for at least 3 years (mean, 3.55 +/- 0.6 years) after diagnosis. Horizontal (MH) and vertical (MV) metamorphopsia scores were obtained with the M-CHARTS. Horizontal and vertical retinal contraction due to ERM was measured by using image-analysis software developed by the authors to calculate horizontal and vertical components of changes in the locations of retinal vessels on sequential fundus images. RESULTS: There was a significant (P < 0.01) positive correlation between the degree of retinal contraction and metamorphopsia score. In addition, there were significant positive correlations between horizontal contraction of the retina and the MV score (P < 0.01) and between vertical contraction of the retina and the MH score (P < 0.05). No significant correlations were found between change in the metamorphopsia score and change in visual acuity or mean defect. CONCLUSIONS: Metamorphopsia scores correlate well with measurements of retinal contraction due to idiopathic ERM. Using M-CHARTS is a simple and useful method for quantitatively monitoring metamorphopsia in patients with ERM. PMID- 16043873 TI - Contrast response properties of magnocellular and parvocellular pathways in retinitis pigmentosa assessed by the visual evoked potential. AB - PURPOSE: To evaluate the contrast response of the visual system in retinitis pigmentosa (RP) under conditions designed to emphasize the parvocellular (PC) and magnocellular (MC) pathways. METHOD: Visual evoked potentials (VEPs) were measured in 10 patients with RP and in 10 age-equivalent control subjects with normal visual acuity and color vision, by using an array of isolated checks that were presented against a steady yellow background. The checks were modulated sinusoidally, either in isoluminant chromatic contrast (5.6 Hz), to favor the chromatic PC pathway, or in luminance contrast (5.6 and 11.2 Hz), to favor the MC pathway. Response amplitude and phase at the stimulus (fundamental) frequency were derived from Fourier analysis, and contrast response functions were fit with a Michaelis-Menten equation to derive R(max), the maximum response amplitude, and sigma, the contrast necessary to produce R(max)/2. RESULTS: In the control subjects, the mean amplitude function for chromatic modulation increased approximately linearly with increasing contrast, whereas the function for luminance modulation increased sharply at low contrasts and saturated at contrasts above approximately 30% for both temporal frequencies, as expected. The patients with RP showed primarily a reduction in R(max) with little change in sigma in all testing conditions. The reduction in R(max) was equivalent for chromatic modulation and luminance modulation at 5.6 Hz, but was substantially lower for luminance modulation at 11.2 Hz. CONCLUSION: Contrast processing was impaired within both the MC and PC pathways in these patients with RP, but the degree of impairment within the MC pathway depended on temporal frequency. These VEP results are in general agreement with recent psychophysical studies of contrast sensitivity losses in patients with RP, and further they characterize contrast processing deficits in these patients at suprathreshold levels. PMID- 16043874 TI - Effects of oxidized and glycated LDL on gene expression in human retinal capillary pericytes. AB - PURPOSE: Modified (oxidized and/or glycated) low-density lipoproteins (LDLs) have been implicated in retinal pericyte loss, one of the major pathologic features of early-stage diabetic retinopathy. To delineate underlying molecular mechanisms, the present study was designed to explore the global effects of modified LDL on pericyte gene expression. METHODS: Quiescent human retinal pericytes were exposed to native LDL (N-LDL), glycated LDL (G-LDL), and heavily oxidized-glycated LDL (HOG-LDL) for 24 hours, and gene expression was evaluated by DNA microarray analysis. Several of the gene responses were checked, and in each case confirmed by reverse-transcription real-time PCR. RESULTS: HOG-LDL induced a gene expression pattern markedly distinct from that of N-LDL or G-LDL, whereas G-LDL elicited gene expression similar to that of N-LDL. A comparison of responses to HOG-LDL versus N-LDL revealed 60 genes with expression that varied by > or =1.7 fold. The HOG-LDL-responsive genes included members of functional pathways, such as fatty acid, eicosanoid, and cholesterol metabolism; fibrinolytic regulation; cell growth and proliferation; cell stress responses; the kinin system; and angiogenesis. CONCLUSIONS: HOG-LDL elicits gene expression in retinal pericytes that may contribute to pericyte loss and other retinal abnormalities in diabetic retinopathy. Observed proapoptotic and proangiogenic responses to HOG-LDL may be of particular importance in this regard. The genes identified through these studies provide potential therapeutic targets for the prevention and treatment of diabetic retinopathy. PMID- 16043875 TI - Retinal-cell-conditioned medium prevents TNF-alpha-induced apoptosis of purified ganglion cells. AB - PURPOSE: Retinal ischemic processes occurring in glaucoma or diabetic retinopathy induce the secretion of tumor necrosis factor (TNF)-alpha. This cytokine was reported to be either toxic to or protective of retinal ganglion cells (RGCs). In the present study, its effect on RGCs was analyzed in different culture conditions. METHODS: Adult rat RGCs were prepared in mixed retinal cell cultures and in purified cultures. They were incubated in normoxic or ischemic conditions, in the presence or absence of TNFalpha and/or conditioned media isolated from rat retinal glial cell cultures and from adult mixed retinal cell cultures. RESULTS: In mixed retinal cell culture, RGCs were insensitive to TNF-alpha, whereas it induced their degeneration in purified adult RGC cultures. This TNFalpha-elicited toxicity was suppressed by TNFalpha-R1-neutralizing antibodies or caspase 8/10 inhibitors. Analyses of mRNA and protein content in purified RGCs revealed a time dependent reduction in the expression of the inhibitor of caspase-8, c-FLIP. c FLIP mRNA was also undetectable after 5 days of culture in the presence of TNFalpha. The retinal cell-conditioned medium protected the RGCs from TNFalpha induced death and prevented the decrease in c-FLIP mRNA and protein in purified cultures. This medium promoted NF-kappaB translocation in purified RGCs, whereas an NF-kappaB inhibitor induced RGC death in mixed retinal cells. CONCLUSIONS: The results confirm that TNFalpha can induce RGC death by TNF-R1 activation. They indicate, however, that other retinal cells can release a molecule that promotes NF-kappaB translocation in RGCs, the synthesis of the anti-caspase-8, c-FLIP, and thereby prevents TNFalpha-mediated RGC death. PMID- 16043876 TI - Identification of sequential events and factors associated with microglial activation, migration, and cytotoxicity in retinal degeneration in rd mice. AB - PURPOSE: To elucidate the role of activated microglia in the photoreceptor apoptosis of rd mice by identifying sequential events and factors associated with microglial activation, migration, and cytotoxicity during retinal degeneration. METHODS: Photoreceptor apoptosis in rd mice at postnatal days (P)8, 10, 12, 14, 16, and 18 was detected by terminal dUTP transferase nick end labeling (TUNEL). Retinal microglia were identified by CD11b antibody. Expression of chemokine mRNA, including monocyte chemoattractant protein (MCP)-1, MCP-3, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, regulated on activation normal T cell expressed and secreted (RANTES), interferon-gamma-inducible 10-kDa protein (IP-10), and fractalkine in the retina were examined by reverse transcription polymerase chain reaction (RT-PCR) assay. Production of tumor necrosis factor (TNF)-alpha in the dystrophic retina was studied by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry analysis. Microglial expression of TNF alpha was determined by double immunolabeling. RESULTS: Whereas photoreceptor apoptosis in the rd mice started at P10 and reached a peak at P16, activation and migration of microglial cells were observed at P10 and peaked at P14. The expression of MCP-1, MCP-3, MIP-1alpha, MIP-1beta, and RANTES transcripts were noted at P8 and reached a peak at P12. Production of TNF-alpha was noted in the outer nuclear layer (ONL) of the rd mice at P8 and reached a peak at P12. At the peak of microglial activity, TNF-alpha was predominantly expressed in the activated microglial cells in the ONL. CONCLUSIONS: Activation of microglia, as well as expression of their signaling molecules (chemokines) and microglia derived toxic factor (TNF-alpha), coincides with or precedes the occurrence of photoreceptor apoptosis, suggesting activated microglia play a major role in retinal degeneration in rd mice. The chemokines MCP-1, MCP-3, MIP-1alpha, MIP 1beta, and RANTES are involved in activation and recruitment of the microglia to the degenerating photoreceptor cell layer. TNF-alpha, produced by the activated microglia, may accentuate the photoreceptor cell death. PMID- 16043877 TI - Identification of P2Y receptor subtypes in human muller glial cells by physiology, single cell RT-PCR, and immunohistochemistry. AB - PURPOSE: Retinal Muller glial cells are known to express metabotropic P2Y receptors. The present study was conducted to identify certain subtypes of P2Y receptors in human Muller cells. METHODS: The patch-clamp technique was used to measure increases of Ca(2+)-dependent K+ currents mediated by the activation of P2Y receptors in freshly isolated human Muller cells. Several P2 agonists were used. Subsequently, the cells were harvested into the patch pipette and a single cell RT-PCR was performed. Moreover, retinal tissue from organ donors was used for immunohistochemistry. RESULTS: The electrophysiological data were consistent with the expression of P2Y1, P2Y2, P2Y4, and P2Y6 receptor subtypes. RT-PCR revealed that mRNA for all these subtypes was present in Muller cells. However, the incidence of P2Y2 receptor mRNA was significantly lower than that of the other subtypes. Immunoreactivity for all four subtypes was found in retinal tissue, partly colocalized with immunoreactivity for vimentin. CONCLUSIONS: The presented data obtained by different techniques revealed that human Muller cells express P2Y1, P2Y2, P2Y4, and P2Y6 receptors. The specific roles of these receptor subtypes in retinal physiology and/or pathophysiology remain to be investigated in future studies. PMID- 16043878 TI - A residual deficit for global motion processing after acuity recovery in deprivation amblyopia. AB - PURPOSE: In a case of bilateral deprivation amblyopia due to congenital cataracts, the global motion sensitivity for stimuli that were well within the passband of the amblyopic visual system were assessed. METHODS: A stochastic global motion stimulus was used, comprising spatially narrow elements with varied spatial frequency, density, contrast, and area distribution. To determine threshold, a two-alternate, forced-choice direction discrimination task was used. RESULTS: There was a selective deficit for global motion processing that was not due to the visibility of the stimuli and was nonselective for spatial scale. The eye with the more complete recovery (acuity 20/20) from pattern deprivation in childhood exhibited the more severe global motion deficit. discussion. The results suggest a primary extrastriate deficit in the dorsal pathway, possibly involving the middle temporal (MT) and the medial superior temporal (MST) cortical areas, that is unrelated to the acuity deficit thought to be in area V1. A similar deficit has recently been shown in strabismic amblyopia. PMID- 16043879 TI - The art of arteriogenesis. AB - The identification of collateral artery growth (arteriogenesis) as the only mechanism to compensate for the loss of an occluded artery forced us to define the mechanisms responsible for this type of vessel growth. To achieve this, a variety of coronary as well as peripheral models of arteriogenesis have been developed. Based on these studies it is obvious that arteriogenesis obeys different mechanisms than angiogenesis, the sprouting of capillaries. Upon occlusion of an artery, the blood flow is redirected into preexisting arteriolar anastomoses that experience increased mechanical forces such as shear stress and circumferential wall stress. The endothelium of the arteriolar connections is then activated, resulting in an increased release of monocyte-attracting proteins as well as an upregulation of adhesion molecules. Upon adherence and extravasation, monocytes promote arteriogenesis by supplying growth factors and cytokines that bind to receptors that are expressed on vascular cells within a limited time frame. Animal studies evidenced that factors, such as monocyte chemoattractant protein-1, granulocyte-monocyte colony-stimulating factor, or transforming growth factor-beta1, that either attract or prolong the lifetime of monocytes efficiently enhance collateral artery growth, an effect that was seen only to a minor degree after application of a single growth factor. Bone marrow derived stems cells and endothelial progenitor cells do not incorporate in growing arteries but, rather, function as supporting cells. Complete elucidation of the mechanisms of arteriogenesis may lead to efficacious therapies counteracting the devastating consequences of vascular occlusive diseases. PMID- 16043880 TI - Acute adaptations of the coronary circulation to exercise. AB - Coronary blood flow is tightly coupled to myocardial oxygen consumption to maintain a consistently high level of myocardial oxygen extraction. This tight coupling has been proposed to depend on periarteriolar oxygen tension, signals released from cardiomyocytes (adenosine acting on K+(ATP) channels), and/or the endothelium (prostanoids, nitric oxide, endothelin [ET]) and autonomic influences (catecholamines), but the contribution of each of these regulatory pathways and their interactions are still incompletely understood. Until recently, experimental studies into the regulation of coronary blood flow during exercise were principally performed in the dog. We have performed several studies on the regulation of vasomotor tone in coronary resistance vessels in chronically instrumented exercising swine. These studies have shown that the coronary resistance vessels in swine lack significant alpha-adrenergic control, but that these vessels are subject to beta-adrenergic feed-forward control during exercise, which is aided by a parasympathetic withdrawal. In addition, withdrawal of an ET-mediated vasoconstrictor influence also contributes to exercise-induced coronary vasodilation. Coronary blood flow regulation by endothelial and metabolic vasodilator pathways contributes to resting vasomotor tone regulation but does not appear to contribute to the exercise-induced coronary vasodilation. Furthermore, blockade of one vasodilator pathway is not compensated by an increased contribution of the other vasodilator mechanisms, suggesting that porcine coronary vasomotor control by endothelial and metabolic factors occurs in a linear additive rather than a nonlinear synergistic fashion. PMID- 16043881 TI - Homocysteine in microvascular endothelial cell barrier permeability. AB - Redox stress activates the endothelium and upregulates matrix metalloproteinases (MMPs), which degrade the matrix and lead to blood-endothelial barrier leakage. Interestingly, elevated levels of plasma homocysteine (Hcy) are associated with vascular dementia, seizure, stroke, and Alzheimer disease. Hcy competes with the gamma-aminobutyric acid (GABA)-A/B receptors and behave like an excitatory neurotransmitter. GABA stimulates the inhibitory neurotransmitter GABA-A/B receptor and decreases arterial blood pressure. However, the neural mechanisms of microvascular remodeling in hyperhomocysteinemia are unclear. This review addresses the idea that Hcy induces microvascular permeability by attenuating the GABA-A/B receptors and increasing redox stress, which activates a disintegrin and metalloproteinase that suppresses tissue inhibitors of metalloproteinase. This process causes disruption of the matrix in the blood-brain barrier. Understanding the mechanism of Hcy-mediated changes in permeability of the blood-brain barrier and extracellular matrix that can alter the neuronal environment in cerebral vascular dementia is of great importance in developing treatments for this disease. PMID- 16043882 TI - Angiotensin AT2 receptor deficiency after myocardial infarction: its effects on cardiac function and fibrosis depend on the stimulus. AB - Hearts of normotensive angiotensin II type 2 receptor (AT2)-deficient mice do not develop fibrosis after angiotensin II-induced chronic hypertension. Thus, the goal of our study was to clarify whether AT2 knockouts (KOs) are also characterized by altered left ventricular (LV) function and modified remodeling of the extracellular matrix (ECM) after induction of myocardial infarction (MI). MI was induced in 5-mo-old female AT2-deficient mice and controls by occlusion of the left coronary artery. Time-matched sham-operated animals served as controls. After 48 h, the first sets of mice were hemodynamically characterized using a pressure-tip catheter (n = 8/group). We also obtained pressure volume loops using a microconductance catheter in additional sets of animals 3 wk after induction of MI (n = 7/group). Finally, the collagen index was illustrated by Sirius red staining and quantified by digital analysis. Whereas the LV function of sham operated animals did not differ between both genotypes, the collagen index was 44% lower in KO animals. Forty-eight hours and 3 wk post-MI, systolic and diastolic LV function were impaired in both AT2-deficient and wild-type (WT) animals to the same extent by approx 45%. No differences were found between the two genotypes with respect to LV hypertrophy and the fibrosis index in the infarcted and noninfarcted areas 3 wk post-MI. While AT2-KO mice had less cardiac collagen content under basal conditions, the receptor deficiency had no significant influence on LV function at the two investigated time points after induction of MI or on the remodeling of ECM at the latter time point. Thus, hypertension-induced fibrosis is probably triggered by other control mechanisms than fibrosis induced by MI. PMID- 16043883 TI - Increased calcium influx mediates increased cardiac stiffness in hyperthyroid rats. AB - Cardiac remodeling (hypertrophy and fibrosis) and an increased left ventricular diastolic stiffness characterize models of hypertension such as the SHR and DOCA salt hypertensive rats. By contrast, hyperthyroidism induces hypertrophy and hypertension, yet collagen expression and deposition is unchanged or decreased, whereas diastolic stiffness is increased. We determined the possible role of increased calcium influx in the development of increased diastolic stiffness in hyperthyroidism by administering verapamil (15 mg/[kg x d] orally) to rats given triiodothyronine (T3) (0.5 mg/[kg x d] subcutaneously for 14 d). Administration of T3 significantly increased body temperature (control: 36.7 +/- 0.2 degrees C; T3: 39.6 +/- 0.2 degrees C), left ventricular wet weight (control: 2.09 +/- 0.02 mg/kg; T3 3.07 +/- 0.07 mg/kg), systolic blood pressure (control: 128 +/- 5 mmHg; T3: 156 +/- 4 mmHg), and left ventricular diastolic stiffness (control: 20.6 +/- 2.0; T3: 28.8 +/- 1.4). Collagen content of the left ventricle was unchanged. Contractile response to noradrenaline in thoracic aortic rings was reduced. Relaxation in response to acetylcholine (ACh) was also reduced in T3-treated rats, whereas sodium nitroprusside response was unchanged. Verapamil treatment of hyperthyroid rats completely prevented the increased diastolic stiffness and systolic blood pressure while attenuating the increased body temperature and left ventricular weight; collagen content remained unchanged. ACh response in thoracic aortic rings was restored by verapamil. Thus, in hyperthyroid rats, an increased calcium influx is a potential mediator of the increased diastolic stiffness independent of changes in collagen. PMID- 16043884 TI - Mitogen-activated protein kinases (p38 and c-Jun NH2-terminal kinase) are differentially regulated during cardiac volume and pressure overload hypertrophy. AB - Chronic pressure overload (PO) and volume overload (VO) result in morphologically and functionally distinct forms of myocardial hypertrophy. However, the molecular mechanism initiating these two types of hypertrophy is not yet understood. Data obtained from different cell types have indicated that the mitogen-activated protein kinases (MAPKs) comprising c-Jun NH2-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 play an important role in transmitting signals of stress stimuli to elicit the cellular response. We tested the hypothesis that early induction of MAPKs differs in two types of overload on the heart and associates with distinct expression of hypertrophic marker genes, namely ANF, alpha-myosin heavy chain (alpha-MHC), and beta-MHC. In rats, VO was induced by aortocaval shunt and PO by constriction of the abdominal aorta. The PO animals were further divided into two groups depending on the severity of the constriction, mild (MPO) and severe pressure overload (SPO), having 35 and 85% aortic constriction, respectively. Early changes in MAPK activity (2-120 min and 1 to 2 d) were analyzed by the in vitro kinase assay using kinase-specific antibodies for p38, JNK, and ERK2. The change in expression of hypertrophy marker genes was examined by Northern blot analysis. In VO hypertrophy, the activity of p38 was markedly increased (10-fold), without changing the activity of ERK and JNK. However, during PO hypertrophy, the activity of JNK was significantly increased (two- to sixfold) and depended on the severity of the load. The activity of p38 was not changed in MPO hypertrophy, whereas it was slightly elevated (50%) in hearts with SPO. Similarly, ERK activity was not changed in hearts with MPO, but a transient rise in activity was observed in hearts with SPO. The expression of ANF and beta-MHC genes was elevated in both PO and VO hypertrophy; however, this change was much greater in hearts subjected to PO than VO hypertrophy. Alpha-MHC expression was downregulated in PO but remained unchanged in VO hypertrophy hearts. Thus, these results demonstrate differential activation of MAPKs in two types of cardiac hypertrophy and this, in part, may contribute to differential expression of cardiac muscle gene expression, giving rise to unique cardiac phenotype associated with different hemodynamic overloads. PMID- 16043885 TI - A new method for targeted drug delivery using polymeric microcapsules: implications for treatment of Crohn's disease. AB - Recent research and clinical evidence suggest that thalidomide could potentially be used to treat inflammation associated with Crohn's disease. However, systemic side effects associated with large doses of this drug have limited its widespread use. Treatment with thalidomide would prove more efficacious if the drug could be delivered directly to target areas in the gut, thereby reducing systemic circulation. Microcapsule encapsulation could enable direct delivery of the drug. To assess the latter, we designed and tested drug-targeting release characteristics of alginate-poly-L-lysine-alginate (APA) microcapsules in simulated gastrointestinal environments. The results show that APA capsules enabled delivery of thalidomide in the middle and distal portions of the small intestine. We also compared the APA membrane formulation with an earlier designed alginate chitosan (AC) membrane thalidomide formulation. The results show that both APA and AC capsules allow for successful delivery of thalidomide in the gut and could prove beneficial in the treatment of Crohn's disease. However, further research is required. PMID- 16043886 TI - Development of cytochrome-c oxidase activity in rat heart: downregulation in newborn rats. AB - Cytochrome-c oxidase (COX) activity of the rat heart was two- to sevenfold activated when the membrane integrity was disrupted by hypotonic swelling, freezing-thawing, or a detergent, indicating that a large portion of COX capacity in intact mitochondria is not active. The effect of detergent was tested in heart mitochondria isolated from 1-, 5-, 15-, 29-, and 60-d-old rats; activation by detergent was up to 20-fold in 1-d-old animals, whereas in mitochondria from 60-d old rats it was only 7-fold. Our data indicate that the rat heart exhibits significant developmental changes dependent on downregulation of COX activity in the neonatal period. PMID- 16043887 TI - The Popeye domain-containing gene family. AB - The Popeye domain-containing gene family has been isolated on the basis of a subtractive screen aiming at the identification of novel genes with a heart restricted gene expression pattern. The gene family codes for membrane proteins containing three transmembrane domains. The carboxy-terminal part of the protein is localized to the cytoplasm and contains a protein domain with high sequence conservation named the Popeye domain. This domain is involved in protein homo dimerization. The gene family is expressed in heart and skeletal muscle cells as well as smooth muscle cells. In addition, Popdc genes are expressed in other cell types such as neuronal cells in restricted areas of the brain, spinal cord, and dorsal root ganglia, and in various epithelial cells. Recently, it has been proposed that Popdc proteins may function as a novel family of adhesion proteins. That the expression pattern has been conserved during evolution and is very similar in all vertebrate classes and also in basal chordates suggests that Popdc proteins play an important role in cardiac and skeletal muscle. PMID- 16043888 TI - Protein kinase C activity and isoform expression during early postnatal development of rat myocardium. AB - Total protein kinase C (PKC) activity, its isoform expression, and concentration and fatty acid (FA) composition of diacylglycerol (DAG) were determined in the left ventricular myocardium of the rat during early postnatal development (d 2, 3, 5, 7, and 10). PKC activity measured by the incorporation of 32P into histone IIIS decreased between d 2 and 10 in the homogenate as well as in cytosolic, membrane (100,000 g), and nuclear-cytoskeletal-myofilament fractions (1000 g). Likewise, the expression of PKC isoforms (alpha, delta, and epsilon) determined by immunoblotting generally declined during the period analyzed, although with a variable pattern. In the membrane and nuclear cytoskeletal myofilament fractions, PKCdelta and PKCepsilon expression decreased markedly by d 3, returning to or close to the d 2 level immediately on d 5. PKCalpha expression in the membrane fraction remained almost unchanged by d 7, declining thereafter. PKCdelta and PKCepsilon were associated predominantly with particulate fractions, whereas PKCalpha was more abundant in the cytosolic fraction. DAG concentration exhibited a significant decline by d 5, consistent with the decrease in maximal PKC activity. The unsaturation index of FA in DAG tended to decrease on d 3 owing to the lowered proportion of all polyunsaturated FA of n-6 and n-3 series. These results demonstrate that the developmental decrease in PKC activity and expression in the rat myocardium is not linear and that subcellular localization of the enzyme exhibits isoform-specific day-by-day changes during the early postnatal period. These changes are compatible with the view that PKC signaling may be involved in the control of a rapid switch of myocardial growth pattern during the first week of life. PMID- 16043889 TI - Proinflammatory cytokines upregulate mRNA expression and secretion of vascular endothelial growth factor in cultured human airway smooth muscle cells. AB - Airflow obstruction in chronic airway disease is associated with airway and pulmonary vascular remodeling, of which the molecular mechanisms are poorly understood. Paracrine actions of angiogenic factors released by resident or infiltrating inflammatory cells following activation by proinflammatory cytokines in diseased airways could play a major role in the airway vascular remodeling process. Here, the proinflammatory cytokines interleukin (IL)-1beta, and tumor necrosis factor (TNF)-alpha were investigated on cell cultures of human airway smooth muscle (ASM) for their effects on mRNA induction and protein release of the angiogenic peptide, vascular endothelial growth factor (VEGF). IL-1beta (0.5 ng/mL) and TNF-alpha (10 ng/mL) each increased VEGF mRNA (3.9 and 1.7 kb) expression in human ASM cells, reaching maximal levels between 16 and 24 and 4 and 8 h, respectively. Both cytokines also induced a time-dependent release of VEGF, which was not associated with increased ASM growth. Preincubation of cells with 1 microM dexamethasone abolished enhanced release of VEGF by TNF-alpha. The data suggest that human ASM cells express and secrete VEGF in response to proinflammatory cytokines and may participate in paracrine inflammatory mechanisms of vascular remodeling in chronic airway disease. PMID- 16043891 TI - Growing applications and advancements in microarray technology and analysis tools. AB - In today's field of genomics, traditional gene-by-gene approach is not adequate to meet the demand of processing information generated from mapping the complex biology of the human genome. More global views of analyzing the magnitude of information are necessary, such as with microarrays. Microarray technology today is rapidly uncovering broad patterns of genetic activity and showing insight into gene functions, processes, and pathways. With the growing technology, imminent knowledge is being generated looking into transcriptional processes and biological mechanisms from many different organisms and phylogeny. Many tools are being developed to assist with the analysis of such high-throughput data, many applications are being utilized by this technology, and the field is growing and expanding rapidly to accommodate the expanding genomics era. PMID- 16043890 TI - Transforming growth factor-beta1 and disorders of the lung. AB - The transforming growth factor (TGF) superfamily encompasses about 30 members in mammals. The effect of TGF-beta subfamily members is exerted and regulated via selective pathways of synthesis and signaling that involve activation of latent TGF-beta, specific and high-affinity binding to cell membrane serine/threonine kinase receptors, activation of intracellular cascades that include Smad molecules and mitogen-activated protein kinases, and regulated termination of the effect by diverse mechanisms including protein degradation and transcriptional activation. Several comprehensive reviews on TGF-beta biology in general and on the role of this cytokine in other diseases have been published recently. In recent years an unexpected role of TGF-beta on lung homeostasis has been revealed. Here, we discuss the contribution of TGF-beta to the pathogenesis of asthma and chronic obstructive pulmonary disease, two common illnesses of the lung, as well as of lymphangioleiomyomatosis, a rare disease in women. The information we collate and integrate places TGF-beta at a pivotal point within complex networks that control lung physiology as well as the physiopathology of these lung diseases. PMID- 16043892 TI - Oxidative stress in pathogenesis of chronic obstructive pulmonary disease: cellular and molecular mechanisms. AB - Chronic obstructive pulmonary disease (COPD) is a slowly progressive condition characterized by airflow limitation, which is largely irreversible. Oxidative stress and inflammation are the major hallmarks of COPD. Reactive oxygen species, either directly or via the formation of lipid peroxidation products, such as 4 hydroxy-2-nonenal and F2-isoprostanes, may play a role in enhancing the inflammation through the activation and phosphorylation of mitogen-activated protein kinases (MAPKs) and redox-sensitive transcription factors such as nuclear factor-kappaB and activator protein-1 in COPD. In addition, activation of the MAPK family leads to the transactivation of transcription factors and coactivators (chromatin remodeling). This eventually results in expression of genes regulating a battery of distinct proinflammatory, antioxidant, and stress response genes. The presence of an oxidative stress has important consequences on several events of lung physiology and for the pathogenesis of COPD. These include increased sequestration of neutrophils in the pulmonary microvasculature, oxidative inactivation of antiproteases and surfactants, hypersecretion of mucus, membrane lipid peroxidation, mitochondrial respiration, alveolar epithelial injury/permeability, breakdown/remodeling of extracellular matrix, and apoptosis. Oxidative stress may have a role in the poor efficacy of corticosteroids in COPD. This review discusses cellular and molecular changes that occur in response to smoking and oxidative stress in the pathogenesis of COPD. PMID- 16043893 TI - Regulation of N-myristoyltransferase by novel inhibitor proteins. AB - N-myristoylation ensures the proper function and intracellular trafficking of proteins. Many proteins involved in a wide variety of signaling, including cellular transformation and oncogenesis, are myristoylated. The myristoylation of proteins is catalyzed by the ubiquitously distributed eukaryotic enzyme N myristoyltransferase (NMT). Previously, we reported that NMT activity is higher in colonic epithelial neoplasms than in normal-appearing colonic tissue and that the increase in NMT activity appears at an early stage in colonic carcinogenesis. Furthermore, we observed that NMT expression is elevated in colorectal and gallbladder carcinoma. In our laboratory, an endogenous NMT inhibitor protein (NIP71) was discovered from bovine brain that inhibited NMT activity in rat colonic tumors. Very recently we have demonstrated that the protein NIP71, which is a potential inhibitor of NMT, is homologous to heat-shock cognate protein (HSC70). In addition, we have discovered that enolase is a potent inhibitor of NMT. Further work may elucidate the role of HSC70 and/or enolase in the regulation of NMT, which may lead to the development of a gene-based therapy of colorectal cancer. The interaction of oncoproteomic and oncogenomic data sets through powerful bioinformatics will yield a comprehensive database of protein properties, which will serve as an invaluable tool for cancer researchers to understand the progress of tumorigenesis. PMID- 16043894 TI - Factors influencing [F-18] 2-fluoro-2-deoxy-D-glucose (F-18 FDG) uptake in melanoma cells: the role of proliferation rate, viability, glucose transporter expression and hexokinase activity. AB - Using human (SK-MEL 23, SK-MEL 24 and G361) and murine (B16) melanoma cell lines, the coregulatory potential of the uptake of the positron emission tomography (PET) tracer, [Fluorine-18] 2-fluoro-2-deoxy-D-glucose (F-18 FDG) has been investigated in relationship to tumor characteristics. Comparative studies among the four melanoma cell lines demonstrated that the lowest FDG uptake in SK-MEL 24 corresponded strongly to the data for DT (population doubling time) and MTT (tetrazolium salt) cell viability as well as hexokinase (HK) activity, but was not related to the glucose transporter 1 (GLUT 1) expression level. Furthermore, the FDG uptake in each melanoma cell line measured by cell cycle kinetics was significantly positively correlated to both the proliferation index (PI=S/G2M phase fractions) and the cell viability, though with one exception relating to the PI of the lowest FDG uptake cell line, SK-MEL 24. No positive correlation was found between the expression of GLUT 1 and FDG uptake in any individual cell line. However, the HK activities in SK-MEL 23 and 24 showed considerable positive relationships with FDG uptake. Our present study suggests that both the proliferation rate and the cell viability of melanoma cells may be key factors for FDG uptake and that HK activity, rather than GLUT 1 expression, seems to be a major factor. PMID- 16043895 TI - Factors influencing [F-18] 2-fluoro-2-deoxy-D-glucose (F-18 FDG) accumulation in melanoma cells: is FDG a substrate of multidrug resistance (MDR)? AB - In order to specify the influence of multidrug-resistance (MDR) on the accumulation of the PET tracer, F-18 FDG ([Fluorine-18] 2-fluoro-2-deoxy-D glucose, in melanoma cells, both the MDR function and expression of two human melanoma cell lines SK-MEL 23 and 24, were evaluated. The effects of MDR modulators on FDG accumulation and efflux were also investigated. A functional analysis using representative MDR fluorescent substrates and inhibitors clarified the following characteristics: 1) SK-MEL 23 possesses a highly active function of MRP, but not P-gp. 2) SK-MEL 24 possesses weak functions of both MRP and P-gp. Western blot analysis using monoclonal antibodies for MDR expression demonstrated an exceedingly high MRP expression of SK-MEL 23 and only slight P-gp and MRP expression of SK-MEL 24, corresponding to the functional data. The efflux inhibition assay using F-18 FDG revealed a considerable retention of FDG in SK MEL 23 in the presence of the MRP inhibitor probenecid. It was also found that the P-gp inhibitor verapamil depressed the FDG efflux of SK-MEL 24. Our present in vitro study suggests that FDG may be a substrate of MDR in some melanoma cells and further MDR may be one of the important factors affecting FDG-PET melanoma imaging. PMID- 16043897 TI - Overexpression of phosphorylated-STAT3 correlated with the invasion and metastasis of cutaneous squamous cell carcinoma. AB - In order to evaluate the possible effects of STAT3 phosphorylation and expression of E-cadherin on metastasis of some human epidermal non-melanoma cutaneous tumors, the expression of phosphorylated STAT3 (p-STAT3) and E-cadherin were analyzed by immunohistochemistry staining in formalin-fixed, paraffin-embedded tissue sections of human cutaneous squamous cell carcinoma (SCC), basal cell carcinoma (BCC) and seborrhoeic keratosis (SK). An immunohistochemistry staining technique was employed to measure the expression of p-STAT3 and E-cadherin protein in 30 cases of cutaneous SCC, 20 cases of BCC, 20 cases of SK, and 20 specimens of normal skin. The results were as follows: 1) p-STAT3 protein was abnormally increased in SCC as compared to normal skin and SK (p<0.001). Expression of p-STAT3 in SCC was also significantly higher than in BCC (p<0.05). 2) Expression of p-STAT3 was higher in poorly-differentiated SCC than in well differentiated ones (p<0.05). The positive rate of the expression of p-STAT3 correlated well with the depth of tumor invasion and with metastasis (p<0.05), but there was no correlation between the positive rate and tumor size. 3) E cadherin was strongly expressed on the cell membranes of normal skin and SK, especially on basal cells. E-cadherin was weakly expressed on cell membranes of SCC and BCC (p<0.001), whereas its expression was significantly lower in SCC than in BCC (p<0.05). In SCC, the intensity of E-cadherin expression was correlated with the extent of tumor differentiation, but there was no correlation between the expression intensity and the depth of tumor invasion or tumor size. 4) There was a negative correlation between the expression intensity of p-STAT3 and E cadherin in SCC (rs=-0.372, p<0.05). We concluded that the overexpression of p STAT3 may have an important role in the development of epidermal tumors. Abnormal activation of STAT3 may be related to metastasis potential in SCC and the simultaneous detection of p-STAT3 and E-cadherin may contribute to predicating the prognosis in SCC. PMID- 16043896 TI - Beneficial effects of tissue inhibitor of metalloproteinases-2 (TIMP-2) on chronic dermatitis. AB - Chronic dermatitis, such as contact dermatitis (CD) or atopic dermatitis (AD), is a longstanding inflammatory skin disease with cutaneous damage such as erosion, ulceration, and lichenification due to itch-induced scratching. The resultant lesion can be considered to be a kind of wound. The tissue inhibitor metalloproteases-2 (TIMP-2) accelerates wound healing by enhancing the proliferation and migration of epidermal keratinocytes and dermal fibroblasts; it is also a physiologic inhibitor of matrix metalloproteinases. The aim of this study was to test the effect of TIMP-2 on chronic dermatitis. NC/Kuj mice were sensitized with Dermatophagoides farinae (Df) extract. Eczema was induced by repeated applications of this mite allergen to the skin of 20 sensitized mice that were maintained under specific pathogen-free conditions. One group of 10 mice was then treated with topical TIMP-2 solution (0.1 ml, 0.5%) for 28 days, and the other with vehicle alone and the effects of TIMP-2 were evaluated macro- and microscopically. The effect on skin barrier function was estimated by measuring transepidermal water loss (TEWL). Scoring of gross skin findings showed that TIMP-2 significantly reduced the severity of eczema (P<0.05) on days 12-28. Histological examination revealed that TIMP-2 treated mice manifested lower degrees of hyperkeratosis, acanthosis, and spongiosis in the epidermis and fewer inflammatory cells in the dermis than vehicle-treated mice. There were significant reductions in the epidermal thickness and dermal inflammatory cells in the TIMP-2 treated animals (P<0.01); their TEWL was significantly decreased on day 28 (P<0.05). Our results suggest that NC/Kuj mice with Df extract-induced chronic eczema may be a useful model for investigating chronic dermatitis, and that TIMP-2 may be a good agent for treating chronic dermatitis as well as chronic ulcers. PMID- 16043898 TI - Is there any association between hepatitis G virus (HGV), other hepatitis viruses (HBV, HCV) and Behcet's disease? AB - Infectious agents, especially viruses, have been implicated in the pathogenesis of Behcet's disease (BD). The aim of this study was to determine whether BD is associated with hepatitis viruses. In this study, the serological markers of hepatitis (HBsAg, anti-HBs, anti-HBc and anti-HCV) and viral nucleic acid (HGV RNA) were studied in the sera of 35 patients, all of whom fulfilled the diagnostic criteria of the International Study Group for BD, and the results were compared with those of 36 healthy controls. The prevalences of HBsAg, anti-HBs, anti-HBc in BD patients were 2.9%, 45.7%, and 31.4%, respectively, which were not significantly different from those in healthy controls. None of the subjects in either group were found to be positive for anti-HCV. HGV-RNA was detected in two patients with BD and in none of the healthy controls. In conclusion, BD does not seem to be associated with hepatitis viral infections including hepatitis B, C, or G. PMID- 16043899 TI - Clinical evaluation of Behcet's disease: a five year follow-up study. AB - The frequencies of clinical features of Behcet's disease vary in different studies according to the diagnostic criteria used in the study, the department where the study is done, and the ethnic origin of patients included in the study. In this study, we aimed to evaluate the influence of the length of the follow-up period of patients on the frequencies of clinical features of Behcet's disease. Out of 231 patients who applied to our dermatology clinic from June of 1991 to October of 1996, only 52 patients that were followed up for at least five years were included in this retrospective study. The baseline characteristics of these 52 patients were almost the same as those of the 179 patients who dropped out. Almost all of the frequencies of clinical features at the end of the follow-up period of five years were higher than those at the time of presentation. The most prominent increases were observed for erythema nodosum (59.6% to 75.0%), papulopustular lesions (84.6% to 94.2%), and large vessel involvement (23.1% to 30.8%). According to our results, it is most likely that studies mainly including mostly patients with long-standing disease will report higher figures for the frequencies of these clinical features of Behcet's disease. PMID- 16043900 TI - Skin tags and atherosclerotic risk factors. AB - An association between skin tags and insulin resistance, obesity, impaired carbohydrate and lipid metabolism has been suggested. However, there still is a need for comprehensive and controlled clinical studies. We aimed to evaluate the atherogenic risk factors in patients with skin tags. Thirty-six patients with skin tags who were admitted to the dermatology department and 22 healthy controls were included in this study. Possible subjects who were taking systemic drugs or who had a systemic disease that may be associated with lipid or carbohydrate metabolism abnormalities were excluded from the study. All the measurements were completed in 26 patients. Standard oral glucose tolerance tests were performed on the patient and control groups. Serum insulin, total cholesterol, triglyceride and HDL-cholesterol levels were measured. LDL-cholesterol and VLDL-cholesterol ratios and HOMA-IR and body mass indices were calculated. The mean levels of body mass index, HOMA-IR, and total cholesterol were significantly higher in patients than in controls. In conclusion, skin tags may not be innocent tumoral proliferations; instead, follow-up of such patients with regard to the development of diseases associated with atherosclerosis may be beneficial. PMID- 16043901 TI - Atelocollagen sponge and recombinant basic fibroblast growth factor combination therapy for resistant wounds with deep cavities. AB - Recent advances in bioengineering have introduced materials that enhance wound healing. Even with such new tools, some deep ulcers surrounded by avascular tissues, including bone, tendon, and fascia, are resistant to various therapies and easily form deep cavities with loss of subcutaneous tissue. Atelocollagen sponges have been used as an artificial dermis to cover full-thickness skin defects. Topical recombinant human basic fibroblast growth factor has been introduced as a growth factor to induce fibroblast proliferation in skin ulcers. We applied these materials in combination in two patients with deep resistant wounds: one with a cavity reaching the mediastinum through a divided sternum and one with deep necrotic wounds caused by electric burns. These wounds did not respond to the topical basic fibroblast growth factor alone. In contrast, the combination therapy closed the wounds rapidly without further surgical treatment. This combination therapy is a potent treatment for resistant wounds with deep cavities. PMID- 16043902 TI - Topical vitamin D3 derivatives in treating hyperkeratotic palmoplantar eczema: a report of five patients. AB - The treatment of hyperkeratotic palmoplantar eczema is notoriously difficult. A considerable number of patients do not or only partially respond to the current treatments such as topical corticosteroids, topical keratolytics, or PUVA therapy. The purpose of this pilot study was to look for an alternative treatment for hyperkeratotic palmoplantar eczema. We treated five patients with topical vitamin D3 derivatives (calcipotriol 50 microg/g and maxacalcitol 25 microg/g ointments). The lesions almost disappeared after 2 to 8 weeks of treatment in four patients and extremely improved with a seven week treatment in one patient. No adverse effect was observed during or after the treatment, and routine laboratory investigations were within normal ranges. When relapses occurred, they responded well to retreatment. Although the study is preliminary, the results suggest that vitamin D3 derivatives offer a safe, effective alternative form of treatment for recalcitrant hyperkeratotic palmoplantar eczema. PMID- 16043903 TI - Childhood bullous pemphigoid: report of three cases. AB - Bullous pemphigoid (BP) is a disorder that rarely occurs in children. We hereby describe three cases of childhood BP aged 2-4 months, which are among the youngest reported in the literature. BP was confirmed by histopathology, direct and indirect immunofluorescence with salt-split skin test and immunoblotting. These patients were successfully treated with systemic corticosteroids with a complete clinical remission. PMID- 16043904 TI - Lymphoepithelioma-like carcinoma of the skin with potential for sweat glandular differentiation. AB - Lymphoepithelioma-like carcinoma of the skin (LELCS) is a cutaneous malignancy with histopathological resemblance to lymphoepithelioma of the nasopharynx. Its histogenesis remains unknown, and few cases showing skin appendage differentiation have been reported to date. We present the case of a 77-year-old Japanese male with an asymptomatic red nodule on his left cheek. Because the histopathological study revealed focal growth of tumor cells lacking connections with the epidermis and marked lymphocytic infiltration surrounding the neoplastic cell nests, the case was diagnosed as LELCS. On immunohistological staining, the neoplastic cells were positive for carcinoembryonic antigen (CEA), epithelial membrane antigen (EMA), multi-cytokeratin (CK), CK6, CK18, and CK19. On the basis of these results, we suggested that skin appendage differentiation, particularly sweat glandular differentiation, was present in this case. PMID- 16043905 TI - Bucillamine-induced toxic epidermal necrolysis and fixed drug eruption. AB - We report two cases of bucillamine-induced bullous reactions with keratinocyte necrosis. The first patient, a 27-year-old woman, developed toxic epidermal necrolysis (TEN) over her whole body after taking bucillamine 300 mg/day for seven days. The second patient, a 63-year-old woman, developed several bullous erythemas on the mucous membranes and legs after taking bucillamine for more than two years. The fixed drug eruptions were diagnosed based on a provocation test in addition to clinical and histopathologic findings. These cases highlight the importance of considering fixed drug eruption as well as TEN in the differential diagnosis of bucillamine-induced bullous drug eruption. PMID- 16043906 TI - Microvenular hemangioma: a rare vascular lesion. PMID- 16043907 TI - Estrogen dermatitis responding to leuprolide acetate. PMID- 16043908 TI - Pityriasis rosea-like eruption due to ergotamine: a case report. PMID- 16043909 TI - Beta2-microglobulin-induced cutaneous amyloidosis in a patient with long-term hemodialysis. PMID- 16043910 TI - A case of severe sclerodermatous chronic graft-versus-host disease: exposure to cement containing abundant silica may have been a relevant factor in scleroderma like skin changes. PMID- 16043911 TI - Noma or necrotizing fasciitis? PMID- 16043912 TI - Topical vitamin D3 is effective in treating senile warts possibly by inducing apoptosis. AB - Vitamin D3 ointments containing active forms of vitamin D3 are widely used to treat inflammatory keratotic dermatoses such as psoriasis. Senile wart or seborrheic keratosis is a benign tumor which occurs mainly in the elderly. It has traditionally been treated with surgical procedures, freezing with liquid nitrogen, or laser therapy. We treated senile warts with topical vitamin D3 ointments (tacalcitol, calcipotriol or maxacalcitol). Out of 116 cases treated for 3 to 12 months, 35 (30.2%) showed complete disappearance or more than an 80% decrease in the volume of the tumor, 54 cases (46.6%) showed a decrease in the volume between 40 and 80%, and no remarkable changes or decreases of less than 40% were seen in 27 cases (23.3%). The tumors faded without any inflammatory changes such as erythema or swelling. An organ culture experiment using senile wart as a material with several concentrations of tacalcitol revealed that tacalcitol induced apoptosis in the tissue. On the other hand, only sporadic apoptotic cells were seen in the controls (p<0.001). Vitamin D3 may affect senile warts by inducing apoptosis. Clearance of senile warts, especially on exposed areas without pain, may improve the quality of life (QOL) of the elderly. PMID- 16043913 TI - Aggressive basal cell carcinoma in the nasal region. AB - It is extremely rare for basal cell carcinoma (BCC) to metastasize, so it is often only simply excised. However, BCC may cause severe local tissue destruction, which often extends to surrounding muscle, cartilage, and bone; it is then termed "aggressive" BCC. We evaluated the safety margin and the reconstruction method in four cases of nasal BCC that were diagnosed as aggressive BCC histopathologically or by imaging, including magnetic resonance imaging (MRI) and computerized tomography (CT) and then treated by excision. The results showed that the larger the aggressive BCC was, the smaller the histopathological safety margins became. The lateral nasal region was classified into three regions, and individual reconstruction was performed according to anatomical unit, resulting in favorable outcomes. Nasal BCC should be closely examinated, it requires a careful treatment strategy similar to that for other malignant skin tumors. PMID- 16043914 TI - Thiersch grafting in recurrent herpes-induced lip depigmentation. AB - Recurrent herpes induced lip-depigmentation is an uncommon condition observed rarely in patients suffering from recurrent herpes labialis. Patients usually belong the age group between 20 and 40 who have had herpes labialis for 4--12 years. These patients do not respond to available methods of treatment such as psoralens and phototherapy because there is reactivation of the virus. Ten cases of recurrent herpes-induced lip depigmentation (HILD) were successfully treated with a combination therapy of acyclovir 400 mg twice a day for one year with Thiersch grafting six months after starting therapy. PMID- 16043915 TI - Narrow-band ultraviolet B phototherapy in patients with psoriasis: for which types of psoriasis is it more effective? AB - Narrow-band ultraviolet B phototherapy with Philips TL-01 lamps is being used increasingly in the treatment of psoriasis. However, the effectiveness of phototherapy with narrow-band ultraviolet B in different clinical types of psoriasis vulgaris has not yet been evaluated in the literature. In the present study, we aimed to determine the efficacy and safety of narrow-band ultraviolet B therapy for different clinical forms of psoriasis vulgaris in our patient population. In this open study, 53 patients with psoriasis vulgaris, skin types II--IV, were enrolled. The patients were classified as guttate, nummular, plaque, or mixed according to their predominant morphological lesions. Evaluation of therapeutic efficacy was performed by comparing their Psoriasis Area and Severity Index (PASI) scores before and after treatment. The cumulative dose and the number of treatments were also calculated. Side effects were noted during the therapy. With respect to the different types of psoriasis, reductions of the PASI were not significantly different (p>0.05). The mean cumulative dose and total sessions in the plaque-type group were greater than those in the guttate and nummular groups (p<0.01, p<0.05). Complete clearing was observed in all patients in the guttate-type group, but the result was not statistically significant (p=0.064). Side effects were reported in five (11%) patients. During the 12 months follow-up period, there were no statistically significant differences in the relapse of the disease among the psoriasis groups (p=0.317). In conclusion, we found that the efficacy of narrow-band ultraviolet B therapy in patients with psoriasis decreased as the predominant lesion size enlarged. Before considering narrow-band ultraviolet B therapy for psoriasis, assessment of lesion size may be useful. Combination therapies may be more effective than phototherapy alone if the patient has predominantly large plaques. PMID- 16043916 TI - Transforming growth factor beta and connective tissue growth factor are involved in the evolution of nevus of Nanta. AB - We report a 59-year-old Japanese male with nevus of Nanta on the face. Histological examination revealed a nest of nevus cells in the dermis and ossification beneath the nevus. Positive staining for transforming growth factor beta and connective tissue growth factor suggested the involvement of these growth factors in the ossification of nevus of Nanta. PMID- 16043917 TI - Poliosis circumscripta associated with neurofibroma. AB - A 32-year-old woman presented at our clinic with a moderately soft, nontender, subcutaneous mass in the left temporal region and white hairs overlying the swelling. The lesion was surgically excised, and there was no recurrence at one year postoperatively. On histological examination, the hair follicles overlying the subcutaneous mass were devoid of pigment and a poorly demarcated fibrocellular process was seen in the subcutaneous mass, which was typical of neurofibroma. To our knowledge, only one case of poliosis associated with neurofibroma has been previously reported. PMID- 16043918 TI - Basal cell carcinoma of the sole. AB - We report a case of a 72-year-old woman with a two year history of a plantar lesion. The clinical appearance, a nodular reddish irregular mass about 2 centimeters in diameter, suggested a tumoral lesion. Histologic examination revealed a basal cell carcinoma. The lesion was excised with no recurrence after two years of follow up. The clinical events, and the histological findings are briefly discussed. PMID- 16043919 TI - Malignant melanoma and acquired dermal melanocytosis on congenital nevus spilus. AB - We reported a case of malignant melanoma and acquired dermal melanocytosis that appeared on congenital nevus spilus; this is the first report from Japan. An 85 year-old woman had had a nevus spilus on the right lower leg since birth. A black brown nodule developed on the nevus three years before treatment. Blue-gray patches were found within the nevus on inspection. Histopathological analysis of these lesions revealed superficial spreading melanoma and acquired dermal melanocytosis, respectively. There have been 19 previous case reports of malignant melanoma on nevus spilus, and there have only been 4 cases of dermal melanocytosis (plaque-type blue nevus) on nevus spilus. We reviewed the reported cases in the literature and discussed the risk factors of nevus spilus. PMID- 16043920 TI - A case of cutaneous myiasis caused by Wohlfahrtia magnifica. AB - Myiasis is caused by the invasion of tissues or organs of man or animals by dipterous larvae. The disease is infrequent in Turkey; it is observed particularly in people with some predisposing factors. A 46-year-old male farmer with nevoid basal cell carcinoma syndrome (NBCCS) presented with the complaint of a blood-tinged discharge and pain in the left frontal-temporal region for three days. Physical examination revealed live maggots in the ulcerous wound resulting from basal cell carcinoma. The larvae were removed with forceps, and the wound was locally dressed with povidone-iodine. The maggots were identified as the third instar larvae of Wohlfahrtia magnifica. PMID- 16043921 TI - Verruciform xanthoma in close association with isolated epidermolytic acanthoma: a case report and review of the Japanese dermatological literature. AB - A 52-year-old man presented to our department with a scrotal skin nodule, first noted as a papule two to three years previously. The nodule was red and pedunculated with a granular surface and a diameter of 10 mm. Three red papules were scattered around the nodule. Histopathologic examination of the nodule showed epidermal papillary hyperplasia, collections of foam cells in the papillary dermis, and a dense infiltration of inflammatory cells into all dermal layers. In addition, granular degeneration was seen in the pedunculated lesion of the nodule free from the foam cells. Microscopic examination of the red papules also showed granular degeneration. The patient was diagnosed with verruciform xanthoma associated with isolated epidermolytic acanthoma. This is the first report of these two lesions occurring at the same site on the scrotum. PMID- 16043922 TI - Heterotopic ossification related to toxic epidermal necrolysis in a patient with Behcet's disease. AB - We describe a 44-year-old woman with an 18-year-history of Behcet's disease (BD) in whom heterotopic ossification (HO) developed as a complication of toxic epidermal necrolysis (TEN). The patient presented with high fever, a progressive erythematous rash including target-like lesions, flaccid blister formations, and severe detachment. The patient was diagnosed with TEN, and methylprednisolone therapy was started. In the 2nd month of her hospital course, painful limitation developed in both her elbow and shoulder joints. Laboratory and radiographic findings confirmed HO of these areas. BD associated with TEN and HO is a previously unreported entity. In such a condition, determination of underlying genetic abnormality is important. Additionally, HO should be considered as a potential cause of the symptoms related to the joints. PMID- 16043923 TI - Amyloidosis cutis dyschromica: a case treated with acitretin. AB - We report a case of amyloidosis cutis dyschromica with diffuse postinflammatory type hypo- and hyperpigmentation all over the body. The diagnosis was confirmed with routine light microscopic and immunohistochemical staining methods. The patient was treated with acitretin with a good response. PMID- 16043924 TI - Congenital hemihypertrophy with hemihypertrichosis. AB - The rare condition in which one side of the body seems to grow faster than the other is called hemihypertrophy. This may be accepted as normal, but there are certain serious conditions that may be associated with this type of growth pattern. Congenital hypertrichosis is believed to be a genetic disorder that is inherited or occurs as a result of spontaneous mutation. The incidence of generalised congenital hypertrichosis is about one in a billion. A seven-month old female case was brought to the hospital with growth and hairiness complaints, which were more apparent in the right side of the face, the right arm and the right leg. Physical examination showed more apparent hypertrophy and hypertrichosis, which covered the entire right side of the body beginning from the face and including the lower extremities and the genitalia. Hypertrichosis was more evident above the right scapula and on the right leg. The diameter of the right upper extremity and the right lower extremity were measured at 13 mm and 14 mm thicker than the left side, respectively. The infant did not have teeth yet and had totally normal nail growth. Skeletal structure, cardiovascular, respiratory, urogenital, central nervous system and eye examinations were all normal. Results of all laboratory analyses, including genetic examination, and those of the imaging techniques were found to be normal. We hereby present the rare co-presence of hemihypertrophy and hemihypertrichosis, for which we could not identify an etiological cause and which is the third such case in the literature. PMID- 16043925 TI - Hypertrophic lupus erythematosus treated successfully with acitretin as monotherapy. AB - Hypertrophic lupus erythematosus (HLE) is a distinct and rare subset of chronic cutaneous lupus erythematosus characterized by verrucous lesions which are chronic in course and resistant to treatment (1). We describe the successful use of acitretin in a patient with HLE who had multiple hyperkeratotic verrucous plaques over the dorsa of his hands, feet, and legs and who failed to respond to local steroids and antimalarials. PMID- 16043926 TI - Palisading neutrophilic granulomatous dermatitis in a Japanese patient with Wegener's granulomatosis. AB - Wegener's granulomatosis (WG) is an etiologically obscure entity with multiple systemic manifestations. Recently, cytoplasmic anti-neutrophil cytoplasmic antibody (C-ANCA) has become recognized as a valuable adjunct in the diagnosis of this disorder. WG typically involves the upper airway, lungs, and kidneys, but any other organ can be involved, including the skin. We encountered a unique case in which a 27-year-old Japanese man with WG presented with various typical cutaneous manifestations. Purpuric skin lesions and erythematous rash on the lower extremities progressively involved and changed into a necrotizing ulceration on his toe. Additionally, several nodules developed on the extensor surfaces of his elbows. His serum C-ANCA level increased remarkably. Leukocytoclastic vasculitis, the most common histopathological finding in WG patients, was detected in a purpuric lesion on his hand. A biopsy of a nodule on his elbow revealed palisading epithelioid histiocyte granulomas with features of leukocytoclastic vasculitis. The distinctive pattern of papules has been referred to as "palisading neutrophilic granulomatous dermatitis". An open lung biopsy confirmed WG with focal necrotizing granuloma. A renal biopsy demonstrated necrotizing vasculitis and crescentic glomerulonephritis. He showed a good response to oral corticosteroids and cyclophospamide with total remission of symptoms. We believe that a careful balance between the clinical manifestations and the histopathological evidence allows for timely treatment of WG, which may prevent serious morbidity or death. Although uncommon, WG can present with various types of accompanying cutaneous lesions. Therefore, clinicians should keep this diagnosis in mind when presented with these manifestations. PMID- 16043927 TI - Erythema nodosum induced by the synergism of acupuncture therapy and flu-like infection. AB - A 32-year-old female patient developed erythema nodosum-like lesions at needle prick sites after acupuncture therapy. Over the next few days, she developed similar new lesions over the extremities, trunk and face along with flu-like symptoms. There were neither genital ulcerations nor eye lesions. A skin biopsy specimen from an extremity lesion showed the characteristic findings of erythema nodosum. Treatment with oral potassium iodide at a dose of 750 mg/day was effective, and there has not been any recurrence to date. We diagnosed this case as erythema nodosum induced by a synergism between acupuncture therapy and a flu like infection. PMID- 16043928 TI - White fibrous papulosis of the neck: a chinese case. AB - A 48-year-old man who presented with papules on his neck was reported. The patient had multiple, discrete, non-follicular papules on the neck that were round-to-oval, 2--3 mm in diameter and pale. They had remained entirely asymptomatic. On histologic examination, there were circumscribed areas of thickened collagen bundles in the papillary to mid-dermis. This clinical and histologic picture suggested the diagnosis of "white fibrous papulosis of the neck". As far as we are aware, this is the first report of the condition from China. PMID- 16043929 TI - The common KRT9 gene mutation in a Japanese patient with epidermolytic palmoplantar keratoderma and knuckle pad-like keratoses. PMID- 16043930 TI - Increased serum levels of interleukin-6, immunoglobulin and acute phase protein in patients with the severe clinical form of inherited epidermolysis bullosa. PMID- 16043931 TI - Lichen striatus following HBV vaccination. PMID- 16043932 TI - Tumid amyloidosis: a novel type of primary localized cutaneous amyloidosis? PMID- 16043933 TI - The changes in circulating levels of vasoactive intestinal polypeptide during exercise and its reproducibility for detection of myocardial ischemia. AB - Vasoactive intestinal polypeptide (VIP) contributes to the regulation of coronary vasomotor tone and circulating levels of VIP have been reported to increase during acute myocardial infarction. However, the changes in VIP concentration during exercise-induced ischemia have not been studied yet. Therefore, we sought to determine whether circulating levels of VIP change during treadmill exercise testing and whether they could be used as a marker of exercise-induced myocardial ischemia. Twenty-nine subjects with definitive positive (group-I) and 20 subjects (group-II) with negative results on treadmill exercise testing were included in this study. In order to assess circulating levels of VIP, blood samples were collected in both groups before exercise, at 5 minutes of exercise, at peak exercise, and at 10 minutes in the recovery period. There were no differences between the two groups with respect to the baseline demographics of age, sex, heart rate, or blood pressure. The metabolic equivalents (METs) values, peak heart rate achieved, peak systolic-diastolic blood pressure, and exercise duration did not differ between the two groups. No significant differences were found in the circulating levels of VIP at any stage of the exercise between the two groups (10.5 +/- 2.5 versus 11.0 +/- 3.5 pmol/L, P = 0.5, 10.6 +/- 2.3 versus 10.6 +/- 3.3 pmol/L, P = 0.9, 10.9 +/- 3.1 versus 11.5 +/- 3.4 pmol/L, P = 0.5, and 10.7 +/- 1.8 versus 11.7 +/- 4.1 pmol/L, P = 0.3, respectively). There was no relationship between the circulating level of VIP and exercise-induced myocardial ischemia, and therefore it could not be used as a marker of exercise-induced myocardial ischemia. PMID- 16043934 TI - Plasma nitric oxide level and its role in slow coronary flow phenomenon. AB - Previous studies have suggested that microvascular abnormalities and endothelial dysfunction cause slow coronary flow (SCF). The objective of this study was to assess the plasma nitric oxide (NO) level and determine its role in the pathogenesis of SCF phenomenon. Thirty-six patients with SCF (group 1) and otherwise patent coronary arteries and 34 subjects with normal coronary flow (group 2) were included in the study. Coronary flow was quantified according to the TIMI Frame Count (TFC) method. Brachial artery endothelium-dependent flow mediated dilatation (FMD) and nitroglycerin (NTG)-induced endothelium-independent dilatation were studied in both groups. In addition, plasma NO levels were measured and their contribution to FMD was determined. The sex, age, body mass index, arterial blood pressure, and heart rate distributions were similar in both groups. TFC was significantly higher in group 1 compared to group 2 for each artery. The plasma NO level was lower in patients with SCF than in control subjects (18.4 +/- 4.4 versus 25.2 +/- 6.3 micromol/L P = 0.001). FMD was significantly smaller in group 1 than in group 2 (4.0 +/- 3.2% versus 10.6 +/- 5.8%, P = 0.0001). The percent NTG-induced dilatation was similar in the two groups (16.8 +/- 1.1% versus 17.1 +/- 1.1%, P = 0.42). In group 1, the plasma NO level was correlated with percent of FMD. Also, the plasma NO level was inversely correlated with TFC for each artery. Reduced NO bioactivity as well as impaired FMD support the presence of endothelial damage in the pathogenesis of SCF phenomenon. PMID- 16043935 TI - Altered patterns of gene expression in response to chronic atrial fibrillation. AB - To obtain greater insight into atrial remodeling at the molecular level we analyzed the changes in gene expression in human atrial tissue between patients with chronic atrial fibrillation (AF) and those with normal sinus rhythm (NSR). cDNA microarray analysis was used to identify genes differentially expressed during sustained AF of more than 6 months (n = 9, mean age, 45 +/- 12, 6 males and 3 females) as compared to those with NSR (n = 9, mean age, 47 +/- 13, 6 males and 3 females). Western blot analysis was performed to confirm the altered gene expression and to establish the changes in protein expression. DNA gel electrophoresis to establish DNA ladder formation, which was associated with apoptosis in response to chronic AF, was performed. Microscopic findings were observed via electron microscopy. In the microarray analysis, out of 8,167 candidate genes, 66 genes showed a significant change in the expression level in the patients with chronic AF, which was in contrast to those with NSR. Among those, 31 genes were consistently down-regulated and 35 up-regulated more than 2 fold. The relative amounts of the Bcl-2 and p27 in the atrial tissue were decreased and angiotensin II type 2 (AT2) receptor and p21 were increased in the patients with chronic AF as compared to those with NSR. The atrial cardiomyocytes in chronic AF showed a prominent DNA ladder, which is a biochemical hallmark of apoptosis. The expression of Bcl-2, AT2 receptor, p21, and p27 were consistent with a significant role in the apoptosis of cardiac myocytes in the patients with chronic AF. PMID- 16043936 TI - Lack of association of genetic polymorphisms in the interleukin-1beta, interleukin-1 receptor antagonist, interleukin-4, and interleukin-10 genes with risk of rheumatic heart disease in Taiwan Chinese. AB - Inflammation and genetics may play a role in the pathogenesis of rheumatic heart disease (RHD). The aim of this study was to test whether interleukin (IL)-1beta, IL-1 receptor antagonist (IL-1Ra), IL-4, or IL-10 gene polymorphisms could be used as markers of susceptibility to or severity of RHD among the Chinese population in Taiwan. A group of 115 patients with RHD diagnosed by echocardiography, and 163 age- and sex-matched normal control subjects were studied. IL-1beta promoter, IL-1beta exon 5, IL-1Ra, IL-4 promoter, IL-4 intron 3, and IL-10 gene polymorphisms were identified by polymerase chain reaction based restriction analysis. There was no significant difference in the distribution of genotypes and allelic frequencies between RHD cases and controls for IL-1beta promoter, IL-1beta exon 5, IL-1Ra, IL-4 promoter, IL-4 intron 3, and IL-10 gene polymorphisms. Further categorization of the RHD patients into mitral valve disease and combined valve disease subgroups also revealed no statistical difference in these gene polymorphisms when compared with controls. These findings suggest that the IL-1beta, IL-1Ra, IL-4, or IL-10 gene polymorphisms are not suitable genetic markers for RHD in Taiwan Chinese. PMID- 16043937 TI - Pathophysiological characteristics and responsiveness to neurohormonal antagonism in idiopathic dilated cardiomyopathy patients with antihepatitis C virus antibody. AB - A high prevalence of hepatitis C virus (HCV) infection has been reported among idiopathic dilated cardiomyopathy (DCM) patients. We examined the prevalence of DCM patients with HCV antibody, and the pathophysiological characteristics and responsiveness to neurohormonal antagonism in DCM with HCV. HCV antibodies were determined in 540 patients with cardiac diseases. In 117 DCM patients, clinicopathologic data were evaluated before and 1 year after angiotensin converting enzyme inhibitor and/or beta-blocker (ACE-inhibitor/BB) administration and their prognosis was followed-up for a mean of 72 +/- 41 months. HCV antibodies were found in 12 of 135 DCM patients (8.9%) and in 37 of 405 patients without DCM (9.1%) (P = NS). At baseline, contrary to DCM without HCV, DCM with HCV was associated (P < 0.05) with greater left ventricular (LV) end-diastolic and end-systolic dimension, LV mass, and myocardial diameter in endomyocardial biopsy, and lower % fractional shortening. By multivariate analysis, HCV infection was independently associated with larger LV end-systolic dimension among DCM patients (P = 0.005). The advanced LV dilatation and hypertrophy in DCM with HCV decreased more in response to the ACE-inhibitor/BB therapy compared to DCM without HCV. There were no differences between DCM patients with and without HCV in survival and cardiac event-free rates. In summary, although HCV infection appears not to be the specific cause of DCM, HCV may enhance ventricular remodeling leading to heart failure among DCM patients. Nevertheless, the advanced ventricular remodeling with HCV was adequately reversed by neurohormonal antagonism, and did not lead to an unfavorable outcome. PMID- 16043938 TI - Predictors of congestive heart failure mortality in elderly people from the general population. AB - Congestive heart failure (CHF) is highly prevalent in the elderly. The aim of this study was to identify the predictors of CHF mortality in patients over 65 years of age who were free of CHF at initial screening. A total of 3,282 elderly subjects were recruited in a population-based frame and 12-year events were recorded. Continuous items were divided into tertiles and for each tertile adjusted the relative risk (RR) with 95% confidence intervals (CI) was derived in both genders from multivariate Cox analysis of CHF mortality. Age > or = 72 years ([RR]: 2.24; 95% CI 1.56 - 3.24), male gender ([RR]: 1.4; 95%CI 1.02 - 1.76), clinical history of coronary artery disease ([RR]: 1.25; 95% CI 1.02 - 1.76), pulse pressure > or = 79 mmHg ([RR]: 1.33; 95% CI 1.03 - 1.87), heart rate > or = 81 bpm ([RR]: 1.32; 95% CI 1.10 - 1.96), atrial fibrillation ([RR]: 1.82; 95% CI 1.18 - 2.81), left ventricular hypertrophy ([RR]: 1.42; 95% CI 1.01 - 2.02), diabetes ([RR]: 1.35; 95% CI 1.02 - 1.78), vital capacity < or = 81% of the theoretical value ([RR]: 2.50; 95% CI 1.88 - 3.32), forced expiratory volume in 1 second < or = 72% of the theoretical value ([RR]: 2.02; 95% CI 1.55 - 2.72) and serum sodium level < or = 139 mmol/L ([RR]: 1.95; 95% CI 1.44 - 2.63) predicted CHF mortality. This model is able to identify elderly people at increased risk of death from CHF. PMID- 16043939 TI - Morning versus evening administration of a calcium channel blocker in combination therapy for essential hypertension by ambulatory blood pressure monitoring analysis. AB - Patients with moderate to severe hypertension may need more than two antihypertensive drugs in combination to achieve ideal blood pressure (BP) control. The purpose of this study was to compare the efficacy and safety of administering the antihypertensive agents either all together in the morning or separately with two agents in the morning and one calcium channel blocker (CCB) in the evening. Twenty-four-hour ambulatory BP monitoring (ABPM) was performed among 15 patients (mean, 59 years) with moderate to severe essential hypertension. All patients received at least 3 antihypertensive drugs for ideal BP control. Two treatment regimens were given to each patient: Regimen 1: All antihypertensive agents were given once a day in the morning; Regimen 2: All antihypertensive agents were given in the morning, except the CCB which was given at 4:00 pm. After receiving regimen 1 for 4 weeks, each patient underwent 24-hour ABPM to analyze the BP control. After the first ABPM, each patient was switched to regimen 2. After 4 weeks of treatment with regimen 2, each patient underwent the second ABPM measurement. The pretreatment mean systolic and diastolic BP were 179.6 +/- 21.7 and 107.4 +/- 19.9 mmHg, respectively. Between the two regimens, there was no significant difference in the mean 24-hour BP (126.1 +/- 5.8/73.3 +/ 3.8 versus 130.2 +/- 6.2/75.1 +/- 4.7 mmHg), daytime BP (128.2 +/- 6.5/75.3 +/- 3.8 versus 132.4 +/- 5.8/77.2 +/- 4.4 mmHg), nighttime BP (125.2 +/- 4.9/72.4 +/- 3.3 versus 130.9 +/- 6.2/73.8 +/- 4.1 mmHg), and 24-hour heart rate (65.1 +/- 3.8 versus 64.2 +/- 3.4 bpm). The circadian BP and heart rate profiles were almost identical between regimen 1 and regimen 2. We conclude that in patients with moderate to severe hypertension treated with at least 3 antihypertensive agents, administering a CCB simultaneously with other antihypertensive agents in the morning or separately in the evening did not affect the 24-hour BP control. PMID- 16043940 TI - The evaluation of left ventricular systolic and diastolic functions in patients with Friedreich ataxia. A pulse tissue Doppler study. AB - Friedreich's ataxia (FRDA), the most common subtype of early onset hereditary ataxia, is an autosomal recessive neurodegenerative disorder caused by unstable GAA expansions. Two-dimensional, pulse, and pulse tissue Doppler echocardiographic examinations were performed on 21 patients with GAA expansion. There was no association between left ventricle ejection fraction, tissue Doppler systolic s wave, and left ventricle diastolic functions examined by pulse and tissue Doppler. The septum thickness of patients with Friedreich's ataxia was significantly increased when compared with that of the control group and wall thickness was found to be associated with GAA repeats. In patients with FRDA, despite a correlation between genetic abnormality with left ventricular early and late diastolic parameters, global diastolic functions were preserved when examined by tissue Doppler. PMID- 16043941 TI - Distinguishable optimal levels of plasma B-type natriuretic peptide in heart failure management based on complicated atrial fibrillation. AB - A B-type natriuretic peptide (BNP)-guided strategy is being widely used as a superior management technique for heart failure (HF). However, the optimal target level of BNP to improve the prognosis of HF in clinical practice remains unclear. Several studies have recently demonstrated that the existence of atrial fibrillation (AF) affects plasma BNP levels. We evaluated the prognostic value of BNP assay for HF management and found the optimal target level under the BNP guided HF management according to the basal cardiac rhythms: AF or sinus rhythm (SR). Patients hospitalized for HF exacerbation between 1996 and 2002 were stratified into SR (n = 129) and chronic AF (CAF, n = 58) groups as basal cardiac rhythms during hospitalization. Cardiac events including death and re-admission for HF exacerbation after discharge were analyzed in relation to the plasma BNP levels at predischarge. Receiver-operating characteristic (ROC) analysis demonstrated that the cut-off values for predischarge BNP, which predict cardiac events at 36 months after discharge, were 125 pg/mL in the SR group and 165 pg/mL in the CAF group. The area under the ROC curve was 0.72 and 0.82, respectively. Stratified subgroup analysis using the Kaplan-Meier method demonstrated that the risk of a cardiac event decreased in a stepwise fashion across a decreasing predischarge BNP range above these cut-off levels, while the minimum decreased risk was recognized at a BNP range below these cut-off levels in each group. In conclusion, the optimal target levels of plasma BNP at predischarge to improve the prognosis of HF should be different and distinguishable depending on with or without AF. PMID- 16043942 TI - Plasma B-type natriuretic peptide levels and risk factors for congestive heart failure in a Japanese general population. AB - This cross-sectional study was performed to establish the rationale for BNP testing for identifying subjects at high risk of congestive heart failure (CHF) in a screening setting. Plasma BNP concentrations were measured in 8,178 community-dwelling residents (mean age, 62 +/- 12 years; 3,194 males). First, in order to determine age- and sex-related reference values for plasma BNP levels, subjects having factors known to influence plasma BNP levels were excluded. The remaining 3,410 subjects were eligible for the reference study. Second, to verify BNP testing for screening for subjects at high risk of CHF, the clinical characteristics of subjects showing abnormally high plasma BNP levels (> or = 97.5 percentile for each age- and sex-specific value of the reference cohort) were examined. In the reference subjects, plasma BNP levels increased with age in both genders, and were higher in women than in men. In the original cohort, age- and sex-specific reference values for high plasma BNP levels were related to the presence of major ECG abnormalities, hypertension, mildly elevated serum creatinine levels, and a history of coronary heart disease. The results of the present study indicate that individuals with high plasma BNP levels in the community have accumulating risk factors for CHF. This suggests that plasma BNP measurement may be a useful screening test for identification of individuals at high risk of CHF within a Japanese general population. PMID- 16043943 TI - Comparison of positive cases for B-type natriuretic peptide and ECG testing for identification of precursor forms of heart failure in an elderly population. AB - Plasma B-type natriuretic peptide (BNP) levels have been reported to be elevated in various types of cardiac disorders and in precursors of CHF. To elucidate the potential ability of BNP testing to identify individuals with structural cardiac disease (ie, hypertensive heart disease, coronary heart disease, valvular heart disease) among community-dwelling elderly persons, cases which were positive on BNP testing were compared to those positive on ECG testing. In the initial phase, we performed plasma BNP measurements and ECG in 856 participants (age > or = 65 years) selected from a general population. From within this group, subjects with an abnormal ECG (n = 125) were selected according to the Minnesota code. Subjects with elevated BNP were selected independently on the basis of plasma levels (n = 112). In the next phase, subjects in both groups were invited to complete Rose's angina questionnaire and to undergo physical examination and transthoracic echocardiography. In this subject group (positive in ECG testing and/or BNP testing), the two tests had comparable sensitivity (65% versus 59%: NS) and specificity (40% versus 41%: NS) for identifying hypertensive heart disease (n = 17). For coronary heart disease (n = 12), the two tests had also comparable sensitivity (58% versus 42%: NS) and specificity (39% versus 41%: NS). However, for selection of valvular heart disease (n = 7), BNP testing had higher sensitivity than ECG testing (100% versus 14%; P < 0.01) with comparable specificity (43% versus 40%: NS). Several types of structural heart disease, in particular valvular heart disease, could be identified exclusively by BNP testing, suggesting that BNP measurement can make a significant contribution to screening for CHF precursors when used in combination with ECG in elderly populations. PMID- 16043944 TI - German origin clusters for high cardiovascular risk in an Italian enclave. AB - Mortality and morbidity appear to be higher in a Cimbrian population representing an enclave of people who migrated from medieval Germany to the secluded Leogra valley in Italy. A population-based study was organized, recruiting 881 elderly subjects of Cimbrian origin and comparing them with a standard control population (SCP, n = 3,282) having comparable general characteristics and lifestyle. Serum lipids and glucose, blood pressure, heart rate, respiratory function, ECG abnormalities, and historical events were used as risk indicators. Age-adjusted systolic and pulse pressure were higher in the Cimbrians than in the SCP, while diastolic blood pressure was comparable. The prevalences of arterial hypertension, isolated systolic hypertension, and pulse hypertension were significantly more represented among Cimbrians than SCP. The prevalences of diabetes, hypercholesterolemia, and hypertriglyceridemia were higher among the former than the latter. The ratio between apolipoproteins B and A1 was also higher, while the HDL fraction was significantly lower in Cimbrians than in the SCP. In Cimbrians, the relative risk (RR) for ischemic heart disease was 1.92 (1.57-2.34) in women, 2.30 (1.54-3.43) in men and 1.03 (1.00-1.06) in women for stroke, 2.43 (1.54-3.83) in men and 1.45 (1.01-1.12) in women for atrial fibrillation, 3.85 (2.83-5.24) in men and 1.39 (1.20-1.60) in women for respiratory disease, 1.97 (1.32-2.94) in men and 6.81 (4.38-10.60) in women for intermittent claudication, and 3.31 (2.44-4.50) in men and 2.30 (1.76-3.01) in women for left ventricular hypertrophy. The subjects living in the secluded Leogra valley are at higher cardiovascular risk than the standard controls. Whether this depends on genetic factors, lifestyle, or both will need to be clarified by further analysis. PMID- 16043945 TI - Comparison of the effect of the ATP-sensitive potassium channel opener YM934 with that of nitroglycerin on impaired coronary circulation in dogs. AB - We compared the effect of an ATP-sensitive potassium channel opener, YM934, with that of nitroglycerin (NTG) on impaired coronary circulation in dogs. Coronary stenosis was produced in 7 dogs by placing a hydraulic occluder around the proximal left circumflex coronary (LCx) artery and abolishing reactive hyperemia to compromise the LCx flow. The following parameters were measured: the aortic pressure, LCx flow velocity, LCx vessel diameter, LCx peripheral pressure, and segment length in the LCx area. Subsequently, we occluded the LCx artery for 15 seconds and measured the recovery-interval (time required for the segment shortening to return to the preocclusion value). The measurements were recorded under three study conditions: (1) at baseline without LCx stenosis; (2) with LCx stenosis under NTG infusion (3 microg/Kg/min); and (3) with LCx stenosis after intravenous administration of YM934 (0.3 microg/kg). The heart rate and aortic pressure were similar under the three study conditions. Mean LCx flow velocity and segment shortening did not significantly change either. However, LCx peripheral pressure decreased after the induction of stenosis (P < 0.05) and showed no response to either NTG or YM934. YM934 administration significantly increased LCx flow in the presence of LCx stenosis, (P < 0.05), whereas NTG infusion did not. YM934 significantly shortened the recovery-interval of the segment shortening after 15-sec LCx occlusion (P < 0.05), whereas NTG did not. These findings suggest that YM934 improves coronary blood flow and prevents myocardial ischemic damage in severely impaired coronary circulation. PMID- 16043946 TI - Rapamycin ameliorates experimental autoimmune myocarditis. AB - Myosin-induced autoimmune myocarditis in rats is a model of human dilated cardiomyopathy. Rapamycin is a potent immunosuppressant and specifically inactivates the mammalian target of rapamycin (mTOR). To examine the role of mTOR in autoimmune myocarditis, we administered rapamycin to rats immunized with cardiac myosin. Phosphorylation of p70 ribosomal S6 kinase 1 (S6K1), a target of mTOR, was increased by 6.9 fold in the heart tissue of myosin immunized rats. Rapamycin (2 mg/kg/day) completely suppressed S6K1 and S6 phosphorylation. The amount of interleukin-1beta, interferon-gamma, interleukin-2, or tumor necrosis factor-alpha mRNA in the heart tissue was markedly increased in myosin-immunized rats, and rapamycin significantly attenuated the cytokine gene expressions. Rapamycin improved the survival of the rats and preserved cardiac function. The plasma level of brain natriuretic peptide increased by 4.7 fold in myosin immunized rats, and rapamycin attenuated the increase in plasma brain natriuretic peptide. The heart weight/tibial length ratio of vehicle-treated myosin-immunized rats was increased by 1.81 +/- 0.06 fold compared with vehicle-treated unimmunized rats, and rapamycin suppressed the increase in heart weight. Rapamycin decreased the cellular infiltration and fibrosis of the myocardium. The amount of phosphorylated S6 was increased in the infiltrating mononuclear cells in vehicle-treated myosin-immunized rats. Rapamycin significantly ameliorated myocardial injury and preserved cardiac function in a rat model of autoimmune myocarditis. PMID- 16043947 TI - A case of sinus of Valsalva-right ventricle fistula without a typical aneurysm, and a single origin of the coronary arteries. AB - Few previous reports have described a sinus of Valsalva fistula without an aneurysm in Japanese patients. A single origin of the coronary arteries is a rare coronary anomaly. We describe a 75-year-old woman with a single origin of the coronary arteries and a sinus of Valsalva fistula without a typical aneurysm. Echocardiography showed turbulent flow from the right coronary sinus of Valsalva to the right ventricle throughout the cardiac cycle. Aortography confirmed the presence of a right coronary sinus of Valsalva-right ventricle shunt jet. Echocardiography and aortography demonstrated that there was no deformity of the sinus of Valsalva. Cardiac catheterization revealed that the left-to-right shunt rate was 29% and the Qp/Qs was 1.41. Aortography and coronary angiography did not identify a right coronary artery originating from the right sinus of Valsalva. Coronary angiography revealed that the right coronary artery arose from the proximal part of the left anterior descending artery and did not detect significant organic stenosis of the coronary artery. She was diagnosed as having a sinus of Valsalva to right ventricle fistula without an aneurysm, and a single origin of the coronary arteries. PMID- 16043948 TI - Successful catheter ablation of a right-sided accessory pathway in a child with interruption of the inferior vena cava and azygos continuation. AB - Azygos continuation is seen in 0.6% of patients with congenital heart disease. The major significance of this anomaly lies in its association with complex cardiac defects and the technical difficulty that can accompany cardiac catheterization. We report our experience with successful catheter ablation of a right posteroseptal concealed accessory pathway in a 6-year-old boy with infrahepatic interruption of the inferior vena cava and azygos continuation. PMID- 16043949 TI - A case of severe heat stroke with abnormal cardiac findings. AB - We document serial changes in the electrocardiogram (ECG) and myocardial markers in a case of severe heat stroke treated with cooling procedures. A 23-year-old comatose male with heat stroke was presented in the emergency room. The condition of the patient was complicated by hepatic failure, rhabdomyolysis, acute renal failure, and cardiac abnormalities. ECG revealed diffuse ST-T elevation; serum levels of myocardial markers were remarkably high and diffuse hypokinesis was observed on the echocardiogram. Cooling procedures, including applying cold vapor to the patient's skin, a gastric lavage with cold water, and an intravenous cold fluid infusion were not successful. Since multiple organ damage (heart, liver, central nervous system, and kidney) was evident, we utilized continuous hemodialysis and hemofiltration, using cold dialysate for efficient cooling. The patient recovered from the multiple organ damage and was removed from the intensive care unit 14 days after the onset. The cardiac abnormalities had normalized within several days without any damage to the myocardium. Q waves were not detected in any lead in the ECG. When interpreting ST-T elevation in the ECG of a heat stroke patient, caution should be used so as to not misdiagnose it as an acute myocardial infarction. PMID- 16043950 TI - A rare case of pulmonary hypertension. As a result of arteriovenous fistula after cardiac surgery. AB - A 19 year-old woman who underwent corrective surgery for an atrial septal defect (ASD) and ventricular septal defect (VSD) 12 years previously presented with pulmonary hypertension. Cardiac catheterization showed a fistula between the right subclavian artery and vena jugularis interna. In this case, a rare example of secondary pulmonary hypertension due to acquired arteriovenous fistula after cardiac surgery is presented. PMID- 16043951 TI - Transradial renal artery angioplasty and stenting in a patient with Leriche syndrome. AB - Percutaneous interventional procedures in the renal arteries are usually performed employing a femoral or brachial vascular access. In contrast, the transradial approach has been established for coronary angiography and angioplasty. We encountered a patient with Leriche syndrome who had renovascular hypertension ascribed to a severe left renal artery stenosis. To stabilize his blood pressure, we made an attempt to relieve the renal artery stenosis with Leriche syndrome by transradial renal artery angioplasty and stenting, using devices for coronary intervention. The procedure was successful without complications or residual stenosis. His hypertension improved with less antihypertensive medications. This case suggests that the radial approach might become an alternative entry site for renal artery interventions. PMID- 16043952 TI - Two female nonsmoker Buerger's disease cases with anticardiolipin autoantibodies and a poor prognosis. AB - We present two female nonsmoker Buerger's disease cases with anticardiolipin autoantibodies and a poor prognosis. One was a 64-year-old female who has had multiple lower and upper extremity amputations, while the other was a 32-year-old female with extremity and visceral artery involvement. Since both were positive for anticardiolipin antibodies, we speculate that Buerger's disease is an autoimmune disorder. PMID- 16043953 TI - Pulmonary arteriovenous malformations, hereditary hemorrhagic telangiectasia, and brain abscess. AB - Hereditary hemorrhagic telangiectasia (HHT) is a systemic angiodysplasia inherited as an autosomal dominant disease. Patients with HHT and pulmonary arteriovenous malformations (PAVMs) are at increased risk for brain abscess (BA), a potentially preventable condition as effective treatment for PAVMs is available. In a center dedicated to HHT, a history of BA was found in 6 out of 128 patients with a definite diagnosis: herewith, their histories are reported focusing on mistakes in the diagnosis and management of the disease. Patients with PAVMs and BA had a higher mean hemoglobin concentration (15.1 g/dl vs. 12.2 g/dl, p < 0.006 by Student's t test) compared to patients with PAVMs alone. Other clinical features (genetics, bacteriology, types of PAVMs, treatments, outcomes) are also discussed. Prompt diagnosis and screening for visceral involvement is pivotal for HHT patients and their relatives. PMID- 16043954 TI - Localized bronchioloalveolar carcinoma with small foci of active fibroblastic proliferation. A report of 2 cases. AB - Localized bronchioloalveolar carcinoma (LBAC) without foci of active fibroblastic proliferation has been reported to be a pN0 disease. Predominantly, these tumors have not been associated with cancer relapse or death subsequent to surgery. Based on these observations, LBAC cases have been considered candidates for limited lung resection without lymph node dissection. However, this surgical option requires careful validation, and an accurate diagnosis is indispensable. The absence of foci of active fibroblastic proliferation must be confirmed by a thorough investigation. Nonetheless, these foci may escape detection, and even currently accepted diagnostic modalities may not be sufficient to accurately determine their absence. We illustrate this point by reporting 2 cases of LBAC with small but distinct foci of active fibroblastic proliferation. PMID- 16043955 TI - Unusual case of pulmonary hypertension. AB - Truly reversible pulmonary hypertension is rare. Acquired systemic arteriovenous (A-V) fistulas following spinal surgery (laminectomy) are a less recognized cause of secondary pulmonary hypertension. We describe a patient who presented with symptoms and clinical evidence of pulmonary hypertension and underwent endovascular correction of an acquired A-V fistula, which led to improvement according to clinical and noninvasive hemodynamic criteria. PMID- 16043956 TI - Radiofrequency thermal ablation of primary pleural synovial sarcoma. AB - Percutaneous radiofrequency (RF) thermal ablation has received attention as an effective minimally invasive approach for the treatment of a variety of neoplasms. However, the therapeutic efficacy of RF thermal ablation on primary pleural synovial sarcoma has not yet been reported. A 76-year-old man with a primary pleural synovial sarcoma who was medically inoperable received RF thermal ablation to achieve local control. Therefore, an RF electrode was inserted into the lesion and connected to an RF generator. In this report, a case of successful treatment of a primary pleural synovial sarcoma using RF ablation without complication is presented. PMID- 16043957 TI - Cor pulmonale and respiratory failure in a young woman. PMID- 16043958 TI - Extralobar pulmonary sequestration in the right upper thoracic region. AB - We present an extremely rare case of extralobar pulmonary sequestration in the right upper thoracic region, with a wide tracheal communication and a right subclavian arterial blood supply. The MRI scan suggested a bronchogenic cyst. Although preoperative color Doppler and angiography were not performed, successful resection was carried out. The histology indicated a diagnosis of extralobar pulmonary sequestration. PMID- 16043959 TI - Pulmonary varix mimicking pulmonary arteriovenous malformation in a patient with Turner syndrome. AB - A 36-year-old asymptomatic female with Turner syndrome was referred for a 3-cm opacity of the left lung detected by routine chest X-ray. A computed tomography scan of the chest suggested a vascular lesion such as pulmonary arteriovenous malformation, and transcatheter embolotherapy was considered. The lack of a right to-left shunt on contrast echocardiography led to suspect an alternate diagnosis. Magnetic resonance imaging and pulmonary angiography eventually demonstrated a pulmonary varix associated with a partial anomalous pulmonary venous return. Contrast echocardiography may help to distinguish between pulmonary varix and arteriovenous malformation. PMID- 16043960 TI - From mirror baby to adolescent. PMID- 16043961 TI - An unusual pulmonary cavitating lesion. PMID- 16043962 TI - Neuroendocrine tumours of the pancreas: predictors of survival after surgical treatment. AB - AIMS: Neuroendocrine tumours of pancreatic and duodenal origin (NETP) are rare and we present a significant experience from a single centre. METHODS: Data was collected on 44 patients who underwent surgery between 1988 and 2002. Since 1997, data have been recorded prospectively on a dedicated database. RESULTS: Twenty four patients had functioning tumours (16 insulinomas, 3 gastrinomas, 2 somatostatinomas, 1 vipoma, 1 glucagonoma and 1 carcinoid tumour). Nine functioning tumours and 13 non-functioning had a malignant phenotype. Twenty pancreaticoduodenectomies, 9 local excisions, 7 distal and 2 total pancreatectomies, 5 bypasses and 1 exploratory laparotomy were performed. Fourteen patients (31.8%) had surgical complications, 1 died peri-operatively (2.3%). The overall actuarial survival for resected cases was 74.4 and 42.5% at 5 and 10 years, respectively. Lymph node invasion and metastases were significant predictors of survival by univariate analysis and only the presence of metastases retained significance on multivariate analysis. CONCLUSION: Surgical resection is the only curative treatment for NETP. Resection can be safely carried out in a specialist centre and is associated with good long-term survival. The presence of metastases was a significant predictive factor for survival in patients with NEPT in this series. PMID- 16043963 TI - Inflammation in atherosclerosis and other conditions: a response to danger. AB - In the last few years atherosclerosis has been recognized as an inflammatory process. Assessment of low-grade inflammation with indexes like C-reactive protein (CRP) is considered indicative and potentially predictive for this disease. On the other hand, in a large number of clinical studies, a grade of microinflammation has been found to be associated with numerous other processes that may be directly, indirectly or not related to atherosclerosis. The most interesting finding these studies have yielded is that innate immunity is activated in various, previously unexpected, conditions. This phenomenon is better explained by the application of a recently proposed immune activation mechanism, namely, the 'danger model'. It seems that many conditions related to metabolic or homeostatic stress or to pro-atherosclerotic and pre-diabetic dysfunctions, established atherosclerosis or diabetes, but also other early tissue injury -- like renal, pulmonary or connective tissue -- and several exogenous stimuli, constitute 'danger signals' that induce inflammation which interacts with and complicates the above-mentioned processes. On this basis the complex relationships between CVD risk factors, atherosclerotic process, other tissue injury and inflammation is examined. The practical application of this hypothesis, namely the new clinical use of CRP as a sensitive -- although not specific for atherosclerosis -- index of low-grade inflammatory activity as well as to the atherosclerosis treatment are also discussed. PMID- 16043964 TI - Prospective comparison of valacyclovir and oral ganciclovir for prevention of cytomegalovirus disease in high-risk renal transplant recipients. AB - AIMS: To compare the efficacy, costs and safety of oral ganciclovir and valacyclovir in the prophylaxis of cytomegalovirus (CMV) disease in renal transplant (RTx) recipients at high risk of CMV disease. METHODS: A total of 83 patients were prospectively randomized to 3-month treatment with either oral ganciclovir (3 g/day) or oral valacyclovir (8 g/day). A 3rd group received no prophylaxis. Forty-nine patients were considered to be at high risk of CMV disease due to D+R- serologic status, OKT3/ATG treatment and/or acute rejection within 12 months after RTx. Twenty-three high-risk patients were treated with ganciclovir (GAN group), 17 patients with valacyclovir (VAL group), and 9 patients received no prophylaxis (C group). RESULTS: No significant differences were found among the groups in their demographic characteristics, immunosuppressive protocols, D/R CMV serology, or CMV risk factors. The 12-month incidence of CMV disease was 89% in the C group compared with 9% in the GAN group and 6% in the VAL group (p < 0.001, GAN or VAL vs. C; p = 0.713, GAN vs. VAL). Treatment failure (death, graft loss, CMV disease or withdrawal from study) occurred in 17, 6, and 89% in the GAN, VAL, and C groups, respectively (p < 0.001, GAN or VAL vs. C; p = 0.285, GAN vs. VAL). The average CMV-associated costs per patient were EUR 3,161, 3,757, and 7,247 in the GAN, VAL, and C groups, respectively (p = 0.027). CONCLUSION: Valacyclovir and oral ganciclovir are equally effective in the prophylaxis of CMV disease in high-risk RTx patients. Both regimens are cost-effective and help reduce CMV-associated costs by nearly 50% compared with patients without prophylaxis. PMID- 16043965 TI - Prevalence of moderate or severe left ventricular diastolic dysfunction in obese persons with obstructive sleep apnea. AB - We investigated prior to gastric bypass surgery the prevalence of left ventricular diastolic dysfunction (LVDD) by Doppler and tissue Doppler echocardiography in 14 obese women and in 6 obese men, mean age 45 years, with a mean body mass index of 49+/-5 kg/m2 who had nocturnal polysomnography for obstructive sleep apnea (OSA). The Doppler and tissue Doppler echocardiographic data were analyzed blindly without knowledge of the clinical characteristics or whether OSA was present or absent. Of 20 patients, 8 (40%) had no OSA, 4 (20%) had mild OSA, and 8 (40%) had moderate or severe OSA. Moderate or severe LVDD was present in 4 of 8 patients (50%) with moderate or severe OSA and in none of 12 patients (0%) with no or mild OSA (p<0.01). Obese patients with moderate or severe OSA have a higher prevalence of moderate or severe LVDD than obese patients with no or mild OSA. PMID- 16043966 TI - Stimulatory effect of interleukin-1beta on growth hormone gene expression and growth hormone release from rat GH3 cells. AB - Our previous studies demonstrated that interferon gamma increases the human (h) growth hormone (GH) gene promoter activity in rat pituitary GH3 cells, and its regulatory mechanism may be different from the classical GH-releasing hormone induced regulatory mechanism. Interleukin-1beta (IL-1beta) is thought to induce the release of GH by pituitary cells, but whether or not and by which mechanisms IL-1beta regulates GH synthesis remains unclear. The purpose of our study was thus to investigate the effect of IL-1beta on the hGH gene expression in GH3 rat pituitary tumor cells using stable transfection of the hGH promoter fused to a luciferase reporter gene. Our results showed that IL-1beta (10-10(4) U/ml) increased GH secretion and synthesis and that 10(2) to 10(4) U/ml IL-1beta promoted the luciferase expression in stable GH3 cells, with a maximal action of 1.61 times over that of controls. Among inhibitors of intracellular signaling transduction pathways, mitogen-activated protein kinase kinase (MAPKK/MEK) inhibitor PD98059 (40 microM) and p38 MAPK inhibitor SB203580 (5 microM)blocked completely the stimulatory effect of IL-1beta, and the phosphoinositide 3-kinase inhibitor LY294002 (10 microM) blocked partially the induction of IL-1beta. Western blot analysis demonstrated that IL-1beta increased the activation of phosphorylated MEK and p38 MAPK in GH3 cells. Neither overexpression of Pit-1 nor inhibiting Pit-1 expression affected IL-1beta induction of hGH promoter activity. To identify the DNA sequence that mediated the effect of IL-1beta, six deletion constructs of hGH promoter were created. The stimulatory effect of IL-1beta was abolished following deletion of the -196- to -132-bp fragment. In conclusion, our data show that IL-1beta promotes GH secretion and synthesis by rat pituitary GH3 cells. The stimulatory effect of IL-1beta on the hGH gene promoter appears to require the activation of MEK, p38 MAPK, and phosphoinositide 3-kinase and a fragment of promoter sequence that spans the -196- to -132-bp fragment of the gene, but is unrelated to the Pit-1 protein. PMID- 16043967 TI - Margatoxin inhibits VEGF-induced hyperpolarization, proliferation and nitric oxide production of human endothelial cells. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) induces proliferation of endothelial cells (EC) in vitro and angiogenesis in vivo. Furthermore, a role of VEGF in K(+) channel, nitric oxide (NO) and Ca(2+) signaling was reported. We examined whether the K(+) channel blocker margatoxin (MTX) influences VEGF induced signaling in human EC. METHODS: Fluorescence imaging was used to analyze changes in the membrane potential (DiBAC), intracellular Ca(2+) (FURA-2) and NO (DAF) levels in cultured human EC derived from human umbilical vein EC (HUVEC). Proliferation of HUVEC was examined by cell counts (CC) and [(3)H]-thymidine incorporation (TI). RESULTS: VEGF (5--50 ng/ml) caused a dose-dependent hyperpolarization of EC, with a maximum at 30 ng/ml (n=30, p<0.05). This effect was completely blocked by MTX (5 micromol/l). VEGF caused an increase in transmembrane Ca(2+) influx (n=30, p<0.05) that was sensitive to MTX and the blocker of transmembrane Ca(2+) entry 2-aminoethoxydiphenyl borate (APB, 100 micromol/l). VEGF-induced NO production was significantly reduced by MTX, APB and a reduction in extracellular Ca(2+) (n=30, p<0.05). HUVEC proliferation, examined by CC and TI, was significantly increased by VEGF and inhibited by MTX (CC: -58%, TI --121%); APB (CC --99%, TI--187%); N-monomethyl-L-arginine (300 micromol/l: CC: -86%, TI --164%). CONCLUSIONS: VEGF caused an MTX-sensitive hyperpolarization which results in an increased transmembrane Ca(2+) entry that is responsible for the effects on endothelial proliferation and NO production. PMID- 16043968 TI - Blood vessels and the aging kidney. AB - Aging is associated with a degenerative effect on many organs including the kidney. Blood vessels play a key role in the progression of renal damage in aging, with reductions in glomerular filtration rate and renal blood flow. Therefore, there is considerable interest in the haemodynamic and molecular mechanisms that may be responsible for alterations in the vascular system in aging. In this review, we will describe the evidence that aging is accompanied by alterations in vascular tone and angiogenesis alongside renal damage. The contributions of mediators such as nitric oxide, angiotensin II and vascular endothelial growth factor will also be discussed. PMID- 16043969 TI - [Atopic dermatitis and skin barrier function]. PMID- 16043970 TI - [Endocrine disruptors and allergy]. PMID- 16043971 TI - [The influence of ageing and seasons on infantile eczema - a population-based cohort study of babies aged 4 months and 10 months]. AB - BACKGROUND: This is probably the first report of a population study on infantile eczema performed for 12 months at two ages of 4 months and 10 months on the same infant group. METHODS: The Fujiidera Health Center services the health needs of 181,994 inhabitants of Habikino and Fujiidera cities. Between September 1990 and August 1991,1775 newborns were called for a health check-up performed twice a months when they reached 4 months of age. This was repeated for a year when the infants reached 10 months of age. A total of 1493 4-month old infants and 1264 10 month old infants were examined. An expert dermatologist joined this and performed a complete skin examination throughout the study. The eczematous skin changes were evaluated at 50 different points on the body and scored using an originally made chart. Scratch marks were also evaluated at each area. Based on the scores computed eczema was diagnosed when the total score crossed a threshold number. Based on this the examiner diagnoses were categorized as follows: Degree 1: no need to visit a doctor, Degree 2: visit to doctor required, Degree 3: treatment required. Gender was not considered. The data was analyzed by an originally made software using DEC-7000 computer. RESULTS: Of all the 4-month old infants examined 329 (22.0%) were initially diagnosed to have eczema while of all the 10-month old infants examined 268 (21.2%) were initially diagnosed to have eczema. Further, out of the 329 4-month old infants initially diagnosed with eczema, 228 were again examined when they were 10-months old. Of this sample; 123 (9.7%) infants showed continued symptoms of eczema and 165 (13.1%) infants no longer showed signs of eczema. In addition, out of the 976 4-month old infants initially diagnosed without eczema, 145 (11.5%) newly developed eczema at 10 months. Hence the cumulative diagnosis rate of eczema reached 34.3%. Meanwhile the monthly diagnosis number was significantly higher in Feb (OR 1.84, p=0.031) and significantly lower in Aug (OR 0.21, p<0.001) than the expected number calculated from monthly examination number and the annual diagnosis rate at 4 months. This observation was also the case at 10 months (Feb; OR 2.19, p=0.02, Aug; OR 0.36, p=0.015). The degree of eczema was significantly higher (p<0.001, Mann-Whitney's U test) at 4 months than 10 months. When this was seen monthly, degree 3 was most prevalent in February-March, while degree 2 was most prevalent in October-January for the 4-month olds. For the 10-month olds, both the degree 3 and degree 2 were quite low throughout the year and only degree 1 showed a monthly change similar to degree 2 plus degree 3 of the 4-month olds. CONCLUSION: We therefore conclude that eczematous skin manifestations of infantile eczema are easily changeable by age at less than 1 year and are strongly influenced by seasons. PMID- 16043972 TI - [Investigation of trends in patients with pollinosis by using web site and the correlation access number and Japanese cedar pollen counts]. AB - BACKGROUND: Recently the number of patient with Japanese cedar pollinosis is increasing. We think many patients are not consulted by a doctor, therefore the questionnaire survey only at hospitals is not enough to know patient's trend. In this study, we examined the trend of patients with pollinosis by the use of a web site. METHODS: For this examination, questionnaire on web site was used from February 1 to April 30, 2003. In this time, the correlation between the pollen counts and access number, how to collect information of pollinosis and the presence of early treatment were focused. RESULTS: The access number to the web site was 348,045 and 1,612 patients answered the questionnaire. There was a good correlation between Japanese cedar pollen counts and the access number to the web site. Most patients (61.2%) collected information about pollinosis and pollen counts by television/radio as well as internet. Patients under 40 years had a medical consultation after the onset symptoms of pollinosis, on the other hand patients over 40 years had a medical consultation before the onset symptoms of pollinosis. Patients who lived in east Japan had a medical consultation before the onset of pollinosis and lived in west Japan after having symptoms. CONCLUSION: It concluded that age and area of patients influenced the patient's trends. Nowadays the spread of internet is remarkable. Therefore, the reliable presentation of information of pollinosis through internet become more important. PMID- 16043973 TI - [Allergenic activity of non-calcinated egg shell calcium]. AB - BACKGROUND: There is few data to prove the safety of egg shell calcium as allergen. Especially, non-calcinated egg shell calcium is not allowed to use without the description of origin of egg on food labels in Japan. METHOD: We evaluated the possibly contaminated egg allergens in commercial non-calcinated egg shell calcium made from egg by in vitro methodologies (SDS-PAGE, western blotting, inhibition ELISA and sandwich ELISA) and by single-blind oral non calcinated egg shell calcium challenge on 6 egg hypersensitivity patients. RESULTS: The allergenic activity of non-calcinated egg shell calcium was almost negligible compared to egg white by all in vitro methods. Furthermore, six patients with egg hypersensitivity were unresponsive to oral non-calcinated egg shell calcium challenge test. CONCLUSION: These results indicate that there is little contamination of egg white in non-calcinated egg shell calcium, and that the allergenic activity of non-calcinated egg shell calcium is equivalent to that of calcinated egg shell calcium. PMID- 16043974 TI - [Clinical aspects of patients with MCS - from the standpoint of allergy]. AB - BACKGROUND: "Sick House Syndrome" is thought to be an illness caused by indoor environments such as allergens, bacteria and chemical compounds. But it is not yet an established clinical entity. "Sick House Syndrome" overlaps in part with Multiple Chemical Sensitivity (MCS) whose symptoms are induced by very small amount of volatile chemical compounds. METHODS: We selected possible cases of MCS from patients who visited our specially built facility for"Sick House Syndrome" by tentative criteria as follow: (1)histories of chemical compounds exposure, (2)multi-organ symptoms, (3)exclusion of other disease(s) which may be responsible for symptoms, (4)chronic symptoms. Clinical aspects of the possible cases were examined. RESULTS: Fifty out of about 130 patients were the possible cases of MCS, 38 females and 12 males, aged 15 to 71 years old. Forty two out of 50 patients (84%) had a history and/or a complication of allergic diseases. This rate is much higher than the rate of prevalence of allergic diseases in Japanese population. Allergic rhinitis was the most popular allergic disease in the possible cases. Total IgE values were relatively low, 32 patients (64%) showed the IgE value below 200 IU/ml. No patients showed anti-formaldehyde IgE antibody. Decreased reactivity and decreased sensitivity of histamine release from peripheral blood were observed after challenge tests with chemical compounds. CONCLUSION: Allergic reactions can not be the causative mechanism(s) of the MCS, which is induced by multiple and different chemical compounds. Our results, however, suggest that patients having allergic diseases may be easily suffered from MCS or MCS may strengthen symptoms of allergic diseases. PMID- 16043975 TI - Critical care challenges in the adult ECMO patient. AB - Extracorporeal membrane oxygenation (ECMO) represents an alternative method of pulmonary support for the critically ill patient with severe respiratory distress. It is commonly used in the neonatal and pediatric populations and is being used with increasing frequency in adults. Although ECMO is not new to the intensive care unit setting, it is usually considered a last resort measure in the adult population. ECMO may save a life and present an awarding challenge to the intensive care unit nurse. PMID- 16043977 TI - Psychosocial interventions for panic disorder after coronary artery bypass graft: a case study. AB - It is increasingly recognized that patients with cardiovascular disease may also suffer from concurrent psychological problems. Many patients present to emergency services and cardiologists with a history of panic disorder. Because of the similarity of presenting symptoms, these patients are often undiagnosed and consequently have slower recovery times and are costly to the healthcare system. Panic disorder is a significant public health problem; however, it is a treatable condition. Healthcare providers should be aware of its occurrence in cardiovascular disease. This case study describes the use of psychosocial interventions, such as the cognitive behavior therapy, in the management of panic disorder after coronary artery bypass graft. A 64-year-old man was treated with 9 sessions of cognitive behavior therapy over a 5-month period. Baseline assessment showed significant distress and deficit in functioning. Following intervention, there was marked reduction in objective and subjective measurement of distress and overall improvement in functioning. Healthcare providers, particularly nurses, need to consider the integration of psychosocial interventions into areas of critical care to provide effective and holistic care. Preoperative screening would be helpful as well. PMID- 16043978 TI - The role of brain natriuretic peptide in systolic heart failure. AB - Heart failure (HF) is a progressive multisystem disease that involves neurohormonal activation, dysfunction of cardiac and skeletal musculature, and a host of other pathological changes. The neurohormonal activation in HF triggers the release of the natriuretic peptides. One peptide of particular interest is brain natriuretic peptide (BNP). It is primarily released by the ventricles of the heart and has adaptive function in counteracting the effects of neurohormonal activation in patients with HF. The focus of this article is the discussion of the physiology of BNP as well as its role in systolic HF, although it also plays a role in diastolic HF. PMID- 16043979 TI - Hiring and mentoring graduate nurses in the intensive care unit. AB - Critical care units must often hire graduate nurses to staff their units. To provide new graduates with the education and experience that they need to independently practice in critical care, mentors are recommended to be assigned to new nurses. These role models help the graduate nurse to grow into the critical care role. PMID- 16043980 TI - Innovative solutions: conferences close to home. AB - Continuing education classes do not have to be taught at far away places to have quality education. This article describes one group's efforts to provide quality continuing education on cardiovascular topics at a local level. PMID- 16043981 TI - Shielding lotions. AB - Unlike traditional moisturizers, shielding lotions help keep out irritants such as latex allergens and antibacterial soaps while locking in the skin's natural moisture. PMID- 16043982 TI - A qualitative examination of the needs of families faced with the option of organ donation. AB - Fewer than half of families approached about organ donation provide consent. Identifying specific support needs of family members in these situations is critical to help them cope and for improving consent rates. This focus group study retrospectively investigated donor and non-donor family members' perceived social support needs while facing the death of their loved one. Implications for nursing care and other interventions are discussed for interested healthcare providers. PMID- 16043983 TI - A closing word: the promise of the future. PMID- 16043984 TI - Correlation of C-reactive protein with clinical, endoscopic, histologic, and radiographic activity in inflammatory bowel disease. AB - INTRODUCTION: We sought to examine the relationship between C-reactive protein (CRP) and clinical, endoscopic, histologic, and radiographic activity in inflammatory bowel disease (IBD). METHODS: All IBD patients at our institution between January 2002 and August 2003 who had a CRP, colonoscopy, and either small bowel follow-through (SBFT) or CT enterography (CTE) performed within 14 days were identified. Clinical activity was assessed retrospectively through review of the medical record. Logistic regression was used in Crohn's disease (CD) patients to estimate the odds ratio (OR) with 95% confidence intervals for an elevated CRP. Associations were assessed using Fisher exact test in ulcerative colitis (UC) patients due to small sample size. RESULTS: One-hundred four CD patients (46% males) and 43 UC and indeterminate colitis patients (44% males) were identified. In CD patients, moderate-severe clinical activity (OR, 4.5; 95% CI, 1.1-18.3), active disease at colonoscopy (OR, 3.5; 95% CI, 1.4-8.9), and histologically severe inflammation (OR, 10.6; 95% CI; 1.1-104) were all significantly associated with CRP elevation. Abnormal small bowel radiographic imaging was not significantly associated with CRP elevation. In UC patients, CRP elevation was significantly associated with severe clinical activity, elevation in sedimentation rate, anemia, hypoalbuminemia, and active disease at ileocolonoscopy, but not with histologic inflammation. CONCLUSIONS: CRP elevation in IBD patients is associated with clinical disease activity, endoscopic inflammation, severely active histologic inflammation (in CD patients), and several other biomarkers of inflammation, but not with radiographic activity. PMID- 16043985 TI - RDP58 is a novel and potentially effective oral therapy for ulcerative colitis. AB - BACKGROUND: RDP58 is a novel anti-inflammatory d-amino acid decapeptide that inhibits synthesis of proinflammatory cytokines by disrupting cell signaling at the pre-MAPK MyD88-IRAK-TRAF6 protein complex. We therefore evaluated its efficacy and safety in parallel multicenter, double-blind, randomized concept studies in ulcerative colitis (UC). METHODS: In the first trial, 34 patients with mild to moderate active UC were randomized (1:2) to placebo (n = 13) or RDP58 100 mg (n = 21). In the second trial, 93 similar patients were randomized (1:1:1) to placebo (n = 30) RDP58 200 mg (n = 31), or RDP 300 mg (n = 32). In both studies, treatment success was defined as a simple clinical colitis activity index score of no more than 3 at 28 days. Sigmoidoscopy and rectal biopsy (at baseline and 28 days) and safety measures (baseline and 28 and 56 days) were other endpoints. RESULTS: Treatment success on RDP 100 mg was 29% versus 46% on placebo (P = 0.46). There were no significant differences in sigmoidoscopy or histology score. In the second study, treatment success on the higher doses of RDP58 (200 and 300 mg) was 71% and 72%, respectively, versus 43% on placebo (P = 0.016). Improvements in sigmoidoscopy scores (41% on 200 mg and 46% on 300 mg versus 32% on placebo) did not reach significance, but histology scores improved significantly (P = 0.002) versus placebo. Overall, adverse events were no different between placebo (3.3 +/- 2.4) and RDP58 (2.7 +/- 1.4, 300-mg group). CONCLUSIONS: RDP58 at a dose of 200 or 300 mg, but not 100 mg, was effective in mild-to-moderate UC. RDP58 was safe and well tolerated, and its novel action makes it an attractive potential therapy. PMID- 16043986 TI - Cytokine expression in healthy and inflamed mucosa: probing the role of eosinophils in the digestive tract. AB - BACKGROUND: In eosinophilic esophagitis (EE), the esophagus is infiltrated with activated eosinophils that often evoke tissue damage, but the intestines of these patients remain unaffected. We thus hypothesized that different tissue-dwelling eosinophil populations may coexist: activated eosinophils that infiltrate the esophagus and resting eosinophils that reside in unaffected intestines. We sought to characterize different eosinophil subpopulations by comparing the expression of certain proinflammatory proteins in tissue-dwelling eosinophils at different parts of the gastrointestinal tract. METHODS: The 8 patients participating included 6 men and 2 women with a previously confirmed diagnosis of EE, whose average age was 39.4 years (range, 20-55 yr) and average disease duration was 13.6 years (range, 2-26 yr). Controls were 3 men and 1 woman, with a mean age of 43.3 years (range, 29-56 yr) with untreated functional dyspepsia who underwent diagnostic esophagogastroduodenoscopy. Six additional individuals having normal blood eosinophils were recruited for cytokine measurements in blood eosinophils. Immunofluorescence and immunoassays charted expression of CD25 and the TH2 cytokines, interleukin (IL)-4, IL-5, IL-10, and IL-13, in esophageal, intestinal, and blood eosinophils from controls and patients. RESULTS: Controls showed a small but significant proportion of intestinal, but no blood, eosinophils expressing CD25 and IL-13, suggesting physiologic activation occurring in the digestive tract. On the other hand, eosinophils infiltrating the inflamed esophageal mucosa of patients with EE showed strong evidence of activation, with most expressing CD25, IL-4, and IL-13. Moreover, IL-13-positive intestinal eosinophils were increased in patients compared with controls. CONCLUSIONS: We thus conclude that tissue-dwelling eosinophils show different and distinct cytokine expression patterns under noninflammatory and inflammatory conditions. PMID- 16043987 TI - Use of endoscopic ultrasound to guide combination medical and surgical therapy for patients with Crohn's perianal fistulas. AB - BACKGROUND: This study was performed to assess if using endoscopic ultrasound (EUS) to assess and guide combination medical and surgical therapy during fistula healing will lead to a high rate of durable fistula closure and a low or absent incidence of perianal abscess formation in patients with Crohn's perianal fistulas. METHODS: This is a retrospective analysis of 21 patients who presented with a symptomatic Crohn's perianal fistula. Patients were enrolled in a clinical practice protocol of serial EUS exams. All patients underwent a baseline rectal EUS and were placed on maximal medical treatment with 6-mercaptopurine (6-MP) or azathioprine, Cipro, and infliximab (5 mg/kg at 0, 2, and 6 wk and then every 8 wk). Patients were also assessed at baseline by a colorectal surgeon who was aware of the EUS findings. Seton placement and incision and drainage were performed when appropriate. Serial EUS examinations were performed, and the findings were used to guide therapy (i.e., the presence of fistula healing on EUS was used to guide seton removal, discontinuation of infliximab, and Cipro). RESULTS: In the 21 patients enrolled, the median duration of active perianal symptoms was 9 wks (1-36). 10 patients (48%) had previous perianal surgery and 5 (24%) had received infliximab previously. The fistulas treated included 8 trans sphincteric, 2 superficial, 3 recto-vaginal, and 7 with multiple and horseshoe fistulas. 13 patients (62%) had associated abscesses at presentation. Eighteen of 21 patients (86%) had complete cessation of drainage initially. Median time to cessation of drainage was 10.6 weeks (range, 4-32 wk). Sixteen of 21 patients (76%) maintained long-term cessation of drainage. The median length of follow-up was 68 weeks (range, 35-101 wk). No abscess developed during treatment in any patient. EUS evidence of persistent fistula activity was seen in 10 patients (48%). Of the 11 patients (52%) in whom EUS showed no persistent fistula activity, 7 (64%) have maintained fistula closure off of infliximab and Cipro. Median duration from last infliximab infusion was 47 weeks (range, 20-80 wk). The remaining 4 patients continued infliximab to maintain remission of their luminal disease. Only 1 patient with a horseshoe fistula showed complete healing on EUS. CONCLUSION: In conclusion, using EUS to guide therapy for Crohn's perianal fistulas with infliximab, an immunosuppressive, and an antibiotic is associated with a high short and long-term fistula response rate. EUS may identify a subset of patients who can discontinue infliximab without recurrence of fistula drainage. PMID- 16043988 TI - Positron emission tomography in the investigation of pediatric inflammatory bowel disease. AB - BACKGROUND: Endoscopic and radiologic studies are frequently required in inflammatory bowel disease (IBD) to determine disease activity, extent of disease, and delineating disease type. Positron emission tomography (PET) using fluorine-18-fluoro-deoxyglucose to identify metabolically active tissues may offer a simple noninvasive alternative to conventional studies in identification and localization of active intestinal inflammation in children with IBD. The aim of this study was to assess the value of PET in identifying active intestinal inflammation compared with conventional endoscopic and radiologic studies, including small bowel follow-through and colonoscopy. METHODS: Sixty-five children were enrolled in the study. This included 55 children (mean age, 13.3 yr; range, 7-18 yr; 20 girls) with newly diagnosed IBD (37) or symptoms suggestive of recurrent disease (18) and 10 children with recurrent abdominal pain (mean age, 12.7 yr; range, 8-15 yr; 7 girls) who were studied with PET, and the results were compared with small bowel follow-through with pneumocolon and/or colonoscopy. Thirty-eight patients had Crohn's disease (17 ileal, 12 ileocolic, 5 pancolonic, 3 left-sided disease, 1 right-sided disease), and 17 had ulcerative colitis (15 pan-colitis, 2 left-sided colitis). Mean time interval between PET and other studies was 30 +/- 17.6 days. RESULTS: PET correctly identified active inflammatory disease in 80% of children with IBD (81.5% with Crohn's disease; 76.4% with ulcerative colitis) and correctly showed no evidence of inflammation in children with recurrent abdominal pain. Gluorine-18-fluoro-deoxyglucose accumulated at sites that corresponded with active disease at colonoscopy in 83.8% of patients and with small bowel follow-through with pneumocolon 75.0% of the time. CONCLUSION: This study suggests that PET offers a noninvasive tool for identifying and localizing active intestinal inflammation in children with IBD. PET may not be able to replace conventional studies; however, it may be useful when conventional studies cannot be performed or fail to be completed. PMID- 16043989 TI - Interleukin-10 polymorphisms in Spanish patients with IBD. AB - BACKGROUND: Inflammatory bowel disease (IBD) is considered a heterogeneous, complex polygenic disease where both genetic and environmental factors are involved in the development of the disease. Interleukin-10 (IL-10) is a regulatory cytokine that might play an important role in disease pathogenesis. IL 10 contains single nucleotide polymorphisms and 2 polymorphic microsatellites in the 5'-flanking region. Our aim was to ascertain if any of these polymorphic markers is associated with IBD among Spanish patients. METHODS: We genotyped 470 patients with IBD, 242 with ulcerative colitis and 228 with Crohn's disease (CD), and 572 ethnically matched controls for microsatellites IL-10R and IL-10G and 2 single nucleotide polymorphisms at positions -1082 and -819 in the proximal promoter of the gene. RESULTS: IL-10G14 microsatellite allele as well as -1082G allele were significantly increased in patients with CD. The combined presence of both alleles in 1 individual notably increased the risk to develop CD (P = 0.00006, odds ratio = 3.18). CONCLUSION: IL-10 polymorphisms contribute to susceptibility to CD in Spanish population. PMID- 16043990 TI - Low-dose oral ferrous fumarate aggravated intestinal inflammation in rats with DSS-induced colitis. AB - BACKGROUND: Oral ferrous iron therapy may reinforce intestinal inflammation. One possible mechanism is by catalyzing the production of reactive oxygen species. We studied the effects of low-dose oral ferrous fumarate on intestinal inflammation and plasma redox status in dextran sulfate sodium (DSS)-induced colitis in rats. METHODS: Forty male Wistar rats were divided into 5 groups: no intervention, sham gavage (distilled water), ferrous fumarate, DSS, and ferrous fumarate + DSS. Ferrous fumarate was dissolved in distilled water (0.60 mg Fe/kg per day) and administered by gavage on days 1 to 14. All rats were fed a standard diet. Colitis was induced by 5% DSS in drinking water on days 8 to 14. Rats were killed on day 16. Histologic colitis scores, fecal granulocyte marker protein, plasma malondialdehyde, plasma antioxidant vitamins, and plasma aminothiols were measured. RESULTS: DSS significantly increased histologic colitis scores (P < 0.001) and fecal granulocyte marker protein (P < 0.01). Ferrous fumarate further increased histologic colitis scores (P < 0.01) in DSS-induced colitis. DSS + ferrous fumarate decreased plasma vitamin A compared with controls (P < 0.01). Otherwise, no changes were seen in plasma malondialdehyde, plasma antioxidant vitamins, or plasma aminothiols. CONCLUSION: Low-dose oral ferrous iron enhanced intestinal inflammation in DSS-induced colitis in rats. PMID- 16043991 TI - Calcaneal ultrasound bone densitometry is not a useful tool to screen patients with inflammatory bowel disease at high risk for metabolic bone disease. AB - BACKGROUND: Up to 42% of patients with inflammatory bowel disease (IBD) have significant metabolic bone disease. The current method of screening for osteopenia or osteoporosis involves dual-energy x-ray absorptiometry (DXA). This is relatively costly and involves radiation exposure. What is needed is a safe, inexpensive, and quick screening tool to identify patients who would benefit from DXA testing. This would reduce the number of patients undergoing DXA testing unnecessarily. We tried to determine if calcaneal ultrasound bone densitometry is a useful tool in screening high-risk patients with IBD for metabolic bone disease. METHODS: Patients with IBD who presented to the clinic between August 29, 2003 and December 22, 2003 were enrolled in this prospective study. All patients underwent calcaneal ultrasound bone densitometry screening using a GE Lunar Achilles Insight quantitative ultrasound densitometry machine (QUS). Patients who were at high risk for significant metabolic bone disease (i.e., significant previous prednisone use or a long history of severe IBD) or who had a T-score on QUS less than or equal to -0.7 had DXA testing performed. The DXA results and QUS results were compared. The radiologist was blinded to the results of QUS. RESULTS: One hundred twenty-four patients with IBD were enrolled. Fifty (40%) were considered high risk for metabolic bone disease. This cohort was comprised of 29 men (58%), of which 21 (73%) had Crohn's disease (CD). Eighty percent of this high-risk group had CD, and in both groups, the majority had used corticosteroids. The overall risk of significant metabolic bone disease in this high-risk group was 62% (DXA < or = -1.0). Heel density (T-score) correlated poorly with DXA (T-score) at either hip or spine at 0.40 even when 2 outlier patients (QUS = -2.9, DXA spine = 0.7, DXA hip = 0.8 and QUS = -3.6, DXA spine = 3, DXA hip = -4) were excluded. Likewise, no association in osteopenia or osteoporosis was seen between multiple variables. These included sex, disease type (ulcerative colitis or CD), smoking, and prior intestinal resection. The sensitivity of QUS to identify patients with significant metabolic bone disease was 74%, and specificity was 63%. A positive predictive value of 81% and negative predictive value of 53% were also less than ideal. The Altman-Bland analysis showed that the agreement between QUS and DXA was poor (-2.0, 2.1). Based on this analysis, QUS cannot replace DXA in the individual patient with IBD. CONCLUSIONS: Calcaneal ultrasound bone densitometry is not a useful tool to screen high-risk patients with IBD for metabolic bone disease. PMID- 16043992 TI - Stepwise regulation of TH1 responses in autoimmunity: IL-12-related cytokines and their receptors. AB - Interleukin (IL)-12 is a key cytokine of cell-mediated immune responses. Until recently, IL-12 was believed to be unique in its ability to induce the differentiation of naive T cells toward the TH1 phenotype and in its pathogenic activity, as shown in various disease models including inflammatory bowel disease. However, recently, 2 additional cytokines closely related to IL-12, IL 23 and IL-27, were discovered. Until then, the role of IL-12 was overestimated because it was believed that the p40 subunit was unique to IL-12. The discovery that IL-12 shares p40 with IL-23 and that IL-23 but not IL-12 is essential in models of chronic inflammation and autoimmunity led to a model in which IL-12 is essential to induce interferon-gamma-producing TH1 cells, whereas IL-23 mediates effector functions. The latest cytokine added to this cytokine family is IL-27. IL-27 has the unique feature to act on naive T cells, rendering them susceptible to IL-12 signaling. Thus, IL-27 may be essential for the early events of a cell mediated immune response. This review focuses on these novel cytokines and their role in cell-mediated immune responses and discusses differences and common features within the family of IL-12-related cytokines. PMID- 16043993 TI - Postoperative Crohn's disease. AB - More than three quarters of patients with Crohn's disease (CD) will require surgery. After resection, disease recurs postoperatively with a median time to second resection of about 10 years. Despite its importance, the postoperative period remains one of the most poorly understood clinical settings in the field. Postoperatively, CD may exhibit unique pathophysiologic features, but the current state of knowledge does not allow for identification of patients at risk for relapse, and leaves clinicians without guidance on optimal maintenance treatment. Therapies used as maintenance for CD in other settings may have different efficacies when used after surgery, and clinical research in patients requiring surgery is limited by the subset of patients available for study. Despite the many limitations in current knowledge of postoperative CD, it is an exciting field because new developments have improved patient care, and ongoing research has the potential for further gains. PMID- 16043994 TI - Autologous hematopoietic stem cell transplantation for Crohn's disease: high risk for a high reward. PMID- 16043995 TI - MRI colonography for IBD: do magnets spin a tale of the inflamed colon? PMID- 16043996 TI - Granulocyte apheresis for pouchitis with arthritis and pyoderma gangrenosum after restorative proctocolectomy for ulcerative colitis: a case report. PMID- 16043997 TI - Lymphoproliferative disorders in collagenous colitis. PMID- 16043998 TI - Lack of evidence for Mycobacterium avium subspecies paratuberculosis in Crohn's disease. PMID- 16043999 TI - Why Trichuris suis should prove safe for use in inflammatory bowel diseases. PMID- 16044000 TI - Effects of HIV-1 infection on lymphocyte phenotypes in blood versus lymph nodes. AB - Most immunopathogenesis studies of HIV-1 use peripheral blood. Most lymphocytes reside in lymphoid tissues, however, and the extent to which blood mirrors tissues is unclear. Here, we analyze lymphocytes in blood and lymph nodes of HIV 1-uninfected and -infected persons. Baseline comparison of node and blood lymphocytes in seronegative persons demonstrates a lower ratio of CD8+ versus CD4+ T lymphocytes, a lower number of effector cells (CD28-) within the CD8+ compartment, and greater activation (D-receptor [DR+]) within the CD4+ compartment. In infected versus uninfected persons, nodes exhibit elevated CD8+ T lymphocytes with an increased memory-effector phenotype (CD62L-/CD45RA-) and activation (CD38+ and DR+) but minimal differences in the CD4+ compartment. Changes attributable to HIV-1 infection are markedly greater in node lymphocytes than in blood. Comparisons of CD8+ T-lymphocyte parameters and viremia in infected persons reveal positive correlations of CD38+ expression on cells in blood and nodes and a negative correlation of terminal effector cells (CD62L /CD45RA+) in the nodes to viremia. Multiple linear regression analysis indicates that CD38 expression on node (not blood) CD8+ T lymphocytes is the sole independent predictor for viremia. Thus, blood indirectly reflects processes in lymphoid tissues, and caution should be applied when interpreting immunopathogenesis studies of blood. PMID- 16044001 TI - Lack of antagonism between abacavir, lamivudine, and tenofovir against wild-type and drug-resistant HIV-1. AB - Potential contributors to the high rate of virologic failure observed for tenofovir, abacavir, and lamivudine include a low genetic barrier to resistance for this regimen and antagonistic drug-drug interactions. To examine the second possibility, we tested combinations of abacavir, tenofovir, and lamivudine against wild-type and drug-resistant HIV-1 in vitro using peripheral blood mononuclear cells and MT-4 cells. Antagonistic interactions were not detected for any combination. If the systems examined accurately reflect the in vivo situation, antagonism does not substantially contribute to the poor efficacy of this triple combination. PMID- 16044002 TI - Highly active antiretroviral therapy (HAART) retreatment in patients on CD4 guided therapy achieved similar virologic suppression compared with patients on continuous HAART: the HIV Netherlands Australia Thailand Research Collaboration 001.4 study. AB - OBJECTIVE: To assess the safety of 2 intermittent treatment strategies compared with continuous therapy for patients with virologic suppression on highly active antiretroviral therapy (HAART) at baseline. DESIGN: Seventy-four nucleoside reverse transcriptase inhibitor (NRTI) and protease inhibitor (PI) pretreated patients with an HIV RNA level <50 copies at screening were randomized to continuous treatment, CD4-guided treatment, or week-on-week-off treatment with 2 NRTIs plus 1600 mg/100 mg of saquinavir/ritonavir once daily. At week 96 (end of the randomized phase of the study), all patients were given continuous HAART for 12 weeks to week 108. Primary outcomes were the proportion of patients with a CD4 count >350 cells/microL and HIV RNA level <400 copies/mL at week 108. METHODS: Patients were followed up every 12 weeks for CD4 count, HIV RNA level, and clinical and laboratory toxicities. In the CD4-guided arm, treatment was stopped and restarted using a CD4 count threshold (above or below 350 cells/microL or reduction of 30%). RESULTS: Seventy-four patients were enrolled with a median CD4 count of 644 cells/microL before the structured treatment interruption (STI). The week-on-week-off arm (n=26) was discontinued at week 72 because of high rates (46%) of HIV RNA rebound above 50 copies/mL. In the continuous arm, 25 (100%) of 25 patients and 24 (96%) of 25 patients had an HIV RNA level <400 copies/mL and <50 copies/mL, respectively, at week 108, and 96% had a CD4 count above 350 cells/microL, with a median CD4 count of 661 cells/microL. Patients in the CD4 guided arm had a significantly lower median CD4 count (489 cells/microL) than the patients in the continuous arm (P=0.03), but all had a CD4 count above 350 cells/microL and 1 had a new HIV-related illness. At week 108, 21 (91%) of 23 patients and 13 (57%) of 23 patients had an HIV RNA level <400 copies/mL and <50 copies/mL, respectively. Those who did not achieve an HIV RNA level <50 copies/mL had a higher HIV RNA load before retreatment, and 4 of 5 patients subsequently achieved viral suppression after an additional 12 weeks of HAART (week 120). Therefore, 17 (94%) of 18 evaluable CD4-guided arm patients achieved viral suppression after retreatment. Antiretroviral (ARV) side effects were similar in all arms. CD4-guided treatment had a 54% ARV cost savings. CONCLUSIONS: This pilot study suggests that CD4-guided HAART is a well-tolerated and cost-saving treatment strategy for patients with high pre-ARV and pre-STI CD4 counts. Week-on week-off treatment had a high virologic failure rate and was discontinued. The HIV RNA suppression rate was similar in patients treated with continuous HAART and in those retreated with 12 to 24 weeks of HAART after CD4-guided therapy. PMID- 16044003 TI - CXCR4 overexpression during the course of HIV-1 infection correlates with the emergence of X4 strains. AB - The factors that determine the emergence of X4 isolates in some HIV-1-infected subjects are unknown. As the level of expression of CXCR4 could favor an R5 to X4 switch, quantitative flow cytometry was used to measure CXCR4 density on CD4 T cells in 200 HIV-1-positive adults, and this was compared with CD4 counts, interleukin-7 (IL-7), and RANTES (regulated on activation, normal T expressed and secreted) plasma levels and the R5/X4 virus phenotype. CD4 T-cell surface CXCR4 densities were increased in infected subjects and inversely correlated with CD4 T cell count (r=-0.548, P<0.001). Yet, in vitro infection with either R5 or X4 strains and in vivo increases in viral load following interruption of antiretroviral treatment did not induce CXCR4 overexpression. The plasma levels of IL-7 and RANTES, 2 cytokines able to induce CXCR4 expression, did not correlate with CXCR4 density. Finally, higher CXCR4 densities were observed in patients harboring X4 strains (3300, 95% CI 2431-4169 CXCR4 molecules per cell) than in patients harboring only R5 strains (2406, 95% CI 2135-2677, P=0.027). These data suggest that CXCR4 overexpression during the course of the disease in some patients could favor the emergence of X4 strains. PMID- 16044004 TI - The association of race, sociodemographic, and behavioral characteristics with response to highly active antiretroviral therapy in women. AB - OBJECTIVE: To determine the association of race with clinical and laboratory outcomes after initiation of highly active antiretroviral therapy (HAART) in HIV 1-infected women in the United States. STUDY DESIGN: Prospective cohort study. PARTICIPANTS: A total of 961 HIV-1-infected women participating in the Women's Interagency HIV Study initiating HAART between July 1, 1995 and September 30, 2003. RESULTS: Over a median of 5.1 years of follow-up, in univariate Cox regression analyses, white women were more likely than African American women to attain a virologic response (relative hazard [RH]=1.34, P=0.005), less likely to experience viral rebound (RH=0.76, P=0.051), and less likely to die (RH=0.63, P=0.040). There were no significant differences, however, among racial groups in outcomes after adjustment for pre-HAART CD4, HIV-1 RNA, history of AIDS-defining illness, age, antiretroviral therapy use, baseline HIV-1 exposure category, and post-HAART behavioral and clinical variables associated with poorer response (discontinuation of HAART, lower income, smoking, current drug use, and depression). Continuous HAART use and lack of depression differed by race and were the strongest predictors of favorable outcomes. CONCLUSION: No significant differences by race were found in virologic, immunologic, or clinical outcomes after adjustment for continued HAART use and depression. These findings suggest that strategies to enhance HAART continuation, including assessing pharmacogenetic influences that may result in greater toxicity and discontinuation rates, and treating depression can improve individual and population-based effects of treatment and potentially mitigate racial disparities in AIDS-related outcomes. PMID- 16044005 TI - The use of community-based modified directly observed therapy for the treatment of HIV-infected persons. AB - Directly observed therapy, which has been successful in the treatment of tuberculosis, is being adapted for the treatment of HIV to decrease long-term morbidity and mortality. We describe the experiences of 69 HIV-infected individuals who were enrolled in a community-based modified directly observed therapy (MDOT) program. Participants were referred by their primary care physicians based on nonadherence to antiretroviral therapy, and/or active substance use. A near-peer outreach worker initially delivered medications to participants 5 to 7 days per week, with visits subsequently tapered to 1 to 3 days per week after 3 or more months. Questionnaires were completed and laboratory values were obtained at baseline, 1 month, and every 3 months after enrollment. At enrollment, 96% of participants had a history of substance use, 71% had a history of incarceration, and 93% were experienced with highly active antiretroviral therapy (HAART). At the time of their 6-month assessment visit, 31 of 69 participants were receiving observed therapy visits. The median baseline plasma viral load (PVL) was 4.8 log, and the median individual change in PVL from baseline to 6 months among participants receiving MDOT was a decrease of 2.7 log. Reasons why participants were not receiving visits included medication holidays, hospitalization or assisted living, incarceration, discontinuation of program involvement, and death. These results support that MDOT should be included in the spectrum of options available to enhance adherence to HAART among patients who are unsuccessful with self-administration of their medications. PMID- 16044006 TI - Therapeutic drug monitoring of efavirenz: trough levels cannot be estimated on the basis of earlier plasma determinations. AB - OBJECTIVE: The few studies evaluating the relation between efavirenz (EFV) plasma levels and efficacy have only shown a weak association. They are based on population pharmacokinetics and/or estimation of trough levels from samples obtained at different time points, because this drug is usually taken at bedtime. We explore whether a limited sampling strategy could provide a good estimation of EFV trough levels and area under the curve (AUC). METHODS: Fifty-nine 24-hour profiles from patients on 600 mg of EFV administered once daily were included for the analysis. Plasma drug concentrations were determined by a validated high performance liquid chromatography assay. The relation between EFV plasma concentrations at C8, C12, and C16 and the corresponding C24 and AUC0-24 values were assessed in different index sets, and their predictive performances were evaluated in the validation sets. RESULTS: EFV plasma levels at C12 and C16 are precise predictors of AUC0-24 (or=30 kg/m) in HIV-infected subjects. RESULTS: Obesity and overweight were more prevalent than wasting (14%, 31%, and 9%, respectively; P<0.0005), but they were not more common than in the general population. Although women and men were equally overweight (30% vs. 31%), women were more obese than men (28% vs. 11%; P<0.001). Among women, African American race (odds ratio [OR]=1.8, 95% confidence interval [CI]: 1.1-2.9) and a CD4 count>or=200 cells/microL (OR=2.8, 95% CI: 1.6-4.9) were associated with overweight and obesity. Among men, only a CD4 count>or=200 cells/microL (OR=1.6, 95% CI: 1.04-2.4) was associated with increased BMI. In men and women, smoking was associated with decreased obesity and overweight (OR=0.59, 95% CI: 0.47-0.74 and OR=0.65, 95% CI: 0.43-0.98, respectively). Age, income, employment, education, past or current intravenous drugs, being on HIV treatment, and viral load were not associated with obesity in the multivariate model. There was a positive correlation between BMI and total cholesterol, triglycerides, and glucose. CONCLUSION: Obesity is more common than wasting in this therapeutic era. Women, particularly those of African American race, are at high risk. Obesity might add to metabolic abnormalities associated with HIV or its treatment and contribute to morbidity, as patients with HIV live longer. PMID- 16044008 TI - HIV antiretroviral treatment: early versus later. AB - OBJECTIVES: Cohort studies indicate that starting highly active antiretroviral therapy (HAART) when the CD4+ T-cell count is less than 200 cells/muL is associated with poor outcomes. These studies have been unable to address how early HAART should be initiated, however. This report uses a modeling approach to compare starting HAART at a mean CD4+ T-cell count greater than 350 cells/microL (early) versus less than 350 cells/microL but greater than 200 cells/microL (later). METHODS: A Markov model tracks people with HIV infection through 6 disease stages defined by CD4+ T-cell count ranges over a 25-year period. Transition probabilities between the disease stages for 6-month periods vary according to initial viral load. Sequences of different first-line, second-line, and "salvage" antiretroviral regimens are defined, and their impact on transition probabilities is estimated. HAART effectiveness is based on data from an urban hospital-based HIV clinic, supplemented by clinical trial data. The model computes the incremental cost-effectiveness of alternative treatment patterns and includes sensitivity analyses for a range of plausible alternative input values. RESULTS: Starting HAART earlier rather than later increases total lifetime costs by $19,074, increases years of life by 1.21 years, increases discounted quality adjusted life-years by 0.61, and has an incremental cost-effectiveness ratio of $31,266 per quality-adjusted life-year. Early therapy is more cost-effective when the impact of HAART on well-being is smaller. CONCLUSIONS: Initiation of HAART at a CD4+ T-cell count greater than 350 cells/microL may be cost-effective (less than $50,000 per quality-adjusted life-year) compared with initiating HAART at a CD4+ T-cell count less than 350 cells/microL but greater than 200 cells/muL and may result in longer quality-adjusted survival. PMID- 16044009 TI - Outbreak of a West African recombinant of HIV-1 in Tashkent, Uzbekistan. AB - OBJECTIVES: This research describes the genetic diversity of HIV-1 in Uzbekistan. METHODS: During 2002 and 2003, blood from HIV-positive patients in Uzbekistan was collected, and part of the proviral pol gene and nearly full-length genomes were sequenced and analyzed. RESULTS: Among 142 Uzbek strains, most clustered genetically with the subtype A strain common in the former Soviet Union. Most of these subtype A-infected drug-naive subjects (65.6%) had an accessory drug resistance mutation, A62V, in the reverse transcriptase gene. Thirteen of the strains (9.2%) clustered with CRF02_AG, an HIV strain common in West Africa. People infected with CRF02_AG were all residents of Tashkent and sampled in 2002. The CRF02_AG strains were monophyletic and probably descended from a single ancestor. Two strains were recombinant between CRF02_AG and subtype A, with each having a different subtype structure. The CRF02_AG and the subtype A elements of the recombinants were monophyletic with Uzbek CRF02_AG and subtype A. New full length genomes of 12 Uzbek strains suggested that neither the subtype A and nor the CRF02_AG strains in this epidemic were mosaics with other subtypes or circulating recombinant forms. CONCLUSION: A genetic analysis of Uzbek HIV strains demonstrated the predominance of subtype A in the epidemic. An outbreak of a West African strain of HIV-1, CRF02_AG, occurred in Tashkent, Uzbekistan in 2002, however. The cocirculation of the 2 strains has resulted in new recombinants that are apparently unique to Uzbekistan. PMID- 16044010 TI - Couples at risk: HIV-1 concordance and discordance among sexual partners receiving voluntary counseling and testing in Uganda. AB - OBJECTIVE: To determine correlates of HIV-1 concordance for couples receiving voluntary HIV counseling and testing. DESIGN: Cross-sectional study of couples receiving voluntary HIV counseling and testing in Kampala, Uganda. METHODS: An interview and physical examination were conducted for 49 HIV-1-concordant (both partners infected with HIV) and 126 HIV-1-discordant (1 partner infected with HIV and 1 partner HIV negative) couples. Blood samples from all participants were tested for HIV-1 and syphilis serology. CD4 cell count and HIV load were characterized for all HIV-infected persons. Urine samples were tested for Neisseria gonorrhoeae and Chlamydia trachomatis using ligase chain reaction. Associations between couples' HIV status and key sociodemographic, behavioral, and biomedical factors were analyzed. RESULTS: Men in HIV-concordant couples were more likely than men in HIV-discordant couples to be living together with their sexual partner (odds ratio [OR], 11.3; 95% confidence interval [CI], 2.8-53.7; P=0.004), to be uncircumcised (OR, 4.5; 95% CI, 1.1-18.8; P=0.042), and to have higher HIV loads (OR for each log increase, 3.0; 95% CI, 2.0-4.7; P<0.001). Women in HIV-concordant couples were more likely than women in HIV-discordant couples to be living together with their sexual partner (OR, 19.0; 95% CI, 3.8-84.8), to have an uncircumcised male partner (OR, 6.5; 95% CI, 1.6-26.4), to have had a sexually transmitted disease in the 6 months before enrollment (OR, 1.9; 95% CI, 0.9-4.5), and to have higher HIV loads (OR for each log increase, 2.2; 95% CI, 1.5-3.2). CONCLUSIONS: Several behavioral and biologic risk factors were associated with HIV concordance for couples. Providing early sexually transmitted disease diagnosis and treatment, antiretroviral therapy, and specially designed counseling to HIV-discordant couples may help prevent HIV transmission in couples where being in a stable sexual relationship is a major risk factor for HIV infection. PMID- 16044011 TI - HIV infection and risk characteristics among female sex workers in Hanoi, Vietnam. AB - The prevalence of HIV/sexually transmitted diseases (STDs) was determined, the risk characteristics examined, and factors associated with HIV infection identified among noninstitutionalized female sex workers (FSWs), using a cross sectional survey with 2-stage cluster sampling. Four hundred FSWs were interviewed face to face using a structured questionnaire and tested for HIV, syphilis, Chlamydia infection, and gonorrhea. HIV seroprevalence was 12%, syphilis 17% (using the treponemal pallidum hemagglutination assay), Chlamydia infection 3.8% (using polymerase chain reaction [PCR]), and gonorrhea 6.3% (PCR). Lower-class FSWs averaged 2 clients per day, and middle-class FSWs about 1.2. Median duration in sex work was 2.3 years. Consistent condom use was 63% with irregular clients, 41% with regular clients, and only 4.8% with "love mates." Fifty-five percent had had sex with a drug user(s). Thirty-eight percent used drugs, of whom 83% injected. Factors associated with HIV included being young, having a low level of education, longer residence in Hanoi, being a lower-class FSW, having higher income compared with peers, perception of self being at low risk for HIV, poor knowledge of HIV, and sharing injecting equipment. Intervention strategies should include reduction of both stigmatization and sharing of drug paraphernalia, promotion of nonstigmatizing voluntary testing and counseling, and aggressive marketing and promotion of condoms. PMID- 16044012 TI - Trends of HIV-1 seroincidence among HIV-1 sentinel surveillance groups in Cambodia, 1999-2002. AB - This study reports trends in HIV-1 incidence in Cambodia among different target groups in the HIV-1 Sentinel Surveillance Program in 1999, 2000, and 2002, using the newly developed IgG capture BED-enzyme (HIV subtypes B, E and D) immunoassay (BED-CEIA). HIV-1-positive specimens (n=3599) from 4 sentinel groups in the HIV-1 Sentinel Surveillance Program from 1999 to 2002-brothel-based commercial sex workers (CSWs), indirect commercial sex workers (IDSWs), police, and women attending antenatal clinics (ANCs)-were tested using the BED-CEIA. Annualized incidence rates were calculated for each group and each geographic region. Between 1999 and 2002, incidence rates declined among CSWs from 13.9% to 6.45%, among IDSWs from 5.92% to 2.87%, and among police from 1.58% to 0.26%. In the ANC group, the incidence remained stable, 0.64% in 1999, 1.11% in 2000, and 0.59% in 2002. However, there was an increasing trend among ANCs in rural areas, from 0.12 to 0.89%. In conclusion, HIV-1 incidence among CSWs, IDSWs, and police has declined between 1999 and 2002; however, the incidence has not declined in the ANC group. PMID- 16044013 TI - Comparison of mother-to-child transmission rates in Ugandan women with subtype A versus D HIV-1 who received single-dose nevirapine prophylaxis: HIV Network For Prevention Trials 012. AB - OBJECTIVE: To compare the rate of mother-to-child transmission (MTCT) in women with subtype A versus D HIV-1 who received single-dose nevirapine (NVP). METHODS: The MTCT rates were compared in women with subtype A versus D at birth and at 8 weeks and 18 months of age of the infants. The rate of late MTCT (after 8 weeks of age) was also analyzed. RESULTS: HIV-1 subtypes were determined for 300 of 306 women who received NVP in the HIV Network for Prevention Trials 012 study (158 women with subtype A and 105 women with subtype D). Infant infection status was known for 297 women. The cumulative rate of MTCT at 18 months was 13.2% for subtype A and 18.3% for subtype D (P=0.34). The rate of late transmission was 3.8% for subtype A and 7.6% for subtype D (P=0.28). Maternal baseline viral load was a significant predictor of MTCT, but maternal baseline CD4 cell count and subtype were not. CONCLUSIONS: No significant difference was observed in the rate of MTCT in women with subtype A versus D. There was a trend toward a higher rate of MTCT among women with subtype D, however, which was also apparent among women whose infants were infected after 8 weeks of age. PMID- 16044014 TI - Predominance of a rare type of HIV-1 in Estonia. AB - An earlier study has indicated that a complex recombinant HIV-1 strain dominates the epidemic in Estonia. The objective of this study was to further investigate the molecular epidemiology and genetic structure of HIV-1 in Estonia. Most of the investigated individuals became infected after August 2000 when HIV-1 started to spread rapidly among Estonian intravenous drug users (IDUs). Two viral DNA regions, gag/pol and gp41, were sequenced and subtyped from peripheral blood mononuclear cells or plasma from 141 individuals. Phylogenetic analysis in the gp41 region revealed that the most frequent type of the virus among IDUs was a circulating recombinant form, CRF06_cpx, whereas a few samples showed highest sequence similarity to a subtype A strain circulating in Ukraine and Russia. Likewise, in the gag/pol region, most of the samples were classified as CRF06_cpx, with a few classified as subtype A. In this region, however, 16% of the sequences turned out to be mosaic unique recombinant forms consisting of CRF06_cpx and subtype A. At least 9 mosaic forms were identified, each with distinct patterns of multiple crossover. To characterize Estonian CRF06_cpx as well as recombinant isolates in more detail, 4 near-full-length HIV-1 genomes were sequenced. PMID- 16044015 TI - Evidence of a brief surge in safer sex practices after HIV testing among a sample of high-risk men and women. AB - OBJECTIVES: We investigated the association between recent HIV testing and safer sex among clients of a publicly funded sexually transmitted disease (STD) clinic. METHODS: Of 401 men and women who were anonymously assessed on sexual risk, HIV testing, STD history, motivations for behavior change, and exposure to HIV prevention education, we studied 292 sexually active respondents who reported previous testing for HIV. Outcome measures included condom use at last intercourse and frequent (at least "most of the time") condom use during the past 3 months. Logistic regression was used to examine the association between each outcome and length of time since HIV testing, controlling for other putative predictors of condom use. RESULTS: The sample consisted mainly of African American (79%) heterosexual (95%) men and women, almost half (44%) of whom were less than 26 years old. Regression analyses indicated that recent HIV testing was significantly associated with safer sex. Prevalence of condom use at last intercourse was highest among respondents who received an HIV test <3 months before the survey, whereas frequent condom use during the past 3 months peaked among clients who had been tested 3 to 5 months before assessment. CONCLUSIONS: The occurrence of a brief "surge" in safer sex among recent HIV counseling and testing clients, regardless of serostatus, suggests that these individuals may be particularly amenable to additional interventions designed to achieve longer term reductions in risky behaviors. PMID- 16044016 TI - Effects of hepatitis C virus coinfection on survival in veterans with HIV treated with highly active antiretroviral therapy. AB - BACKGROUND: With highly active antiretroviral therapy (HAART) available for patients with HIV, hepatitis C virus (HCV) infection has emerged as a potentially important cause of mortality in coinfected patients. Several studies have investigated the effect of coinfection on mortality, with conflicting results. METHODS: The study cohort consisted of HIV-infected veterans on HAART receiving care at US Department of Veterans Affairs facilities. Inclusion was based on first HAART prescription between January 1997 and February 2003, HCV antibody test result, and baseline CD4 and HIV viral load results within 1 year of starting HAART. We fitted Cox proportional hazards models to study the effect of HCV serostatus on survival time from HAART initiation, controlling for patient demographic and clinical characteristics, facility characteristics, HAART exposure, HAART response, and HCV treatment. RESULTS: Of 12,216 patients in the study cohort, 38% were HCV-seropositive. During an observation time averaging 3.5 years, 2087 patients died. The adjusted hazard ratio for HCV-seropositive patients was 1.56 (95% confidence interval [CI]: 1.42-1.70; P<0.0001) without a HAART exposure measure and 1.38 (95% CI: 1.26-1.51; P<0.0001) with the measure. We obtained similar results in analyses also controlling for HAART response. CONCLUSIONS: HCV seropositivity was independently associated with increased risk of death in a large cohort of HAART-treated HIV-infected veterans. Given the success of HAART in extending the lives of HIV patients, HCV has become an important predictor of their mortality. PMID- 16044017 TI - Use of total lymphocyte count and hemoglobin concentration for monitoring progression of HIV infection. AB - BACKGROUND: Prognostic markers for HIV monitoring are needed for resource-limited regions. Prior research has demonstrated rapid declines in total lymphocyte count (TLC) and hemoglobin levels before AIDS, but the prognostic accuracy of these declines has not been examined prospectively. METHODS: Longitudinal TLC and hemoglobin data from men in the Multicenter AIDS Cohort Study (MACS) before the introduction of potent HIV therapy were used to identify the first time when the TLC was 33% per year, and hemoglobin declined by >11.6% per year. The prognostic value of these declines for AIDS was evaluated by Cox regression models and Kaplan-Meier survival curves. RESULTS: Rapid declines in TLC or hemoglobin were associated with progression to AIDS (relative hazard [RH]=4.70, 95% confidence interval [CI]: 3.23-6.86 for TLC; RH=5.55, 95% CI: 3.69-8.36 for hemoglobin). The World Health Organization criterion for initiating therapy, a TLC1200 cells/mm, a rapid decline in TLC or hemoglobin was strongly associated with progression to AIDS (RH=2.53, 95% CI: 1.56-4.10 for TLC; RH=5.28, 95% CI: 3.11-8.97 for hemoglobin). CONCLUSIONS: In the MACS, rapid declines in TLC or hemoglobin concentration indicated an increased likelihood of progression of HIV infection to AIDS. These results support the potential utility of these markers for monitoring HIV-infected people in resource-limited regions, but critical levels and rates of decline of markers for such regions remain to be defined. PMID- 16044018 TI - A community-based study of the incidence of trimethoprim-sulfamethoxazole preventable infections in Malawian adults living with HIV. AB - The benefits of trimethoprim-sulfamethoxazole (TS) prophylaxis reported for persons living with HIV in Cote d'Ivoire are difficult to extrapolate to sub Saharan African countries where bacterial resistance to TS is higher and cross resistance between TS and sulfadoxine-pyrimethamine (SP) may impair SP efficacy for malaria treatment. We conducted a community-based cohort study to measure the incidence of potentially TS-preventable illnesses in Blantyre, Malawi. We found a high incidence of malaria, invasive bacterial infections, and probable bacterial pneumonias but low rates of Pneumocystis jiroveci pneumonia, isosporiasis, and Toxoplasma encephalitis. Most bacterial isolates were resistant to TS but sensitive to azithromycin, a possible alternative to TS. Clinical trials are needed to determine the role of TS or alternative regimens for prophylaxis against secondary infections among people living with HIV in sub-Saharan Africa. These should also assess benefit in patients receiving antiretroviral therapy. PMID- 16044019 TI - Longitudinal assessment of HIV-1 and HSV-2 shedding in the genital tract of West African women. AB - Forty-two HSV-2-seropositive women, of whom 22 were HIV-1 seropositive, were followed up weekly for 3 weeks for assessment of HIV-1 and HSV-2 genital shedding in Bobo Dioulasso, Burkina Faso. HSV-2 DNA and HIV-1 RNA were quantitated in cervicovaginal lavages enriched with a cervical swabbing (eCVL) using real-time polymerase chain reaction. HIV-1 RNA shedding was detected in eCVL from 19 of 22 (86.4%) of the HIV-1-seropositive women and HSV-2 DNA was detected in eCVL from 12 of 42 (28.6%) of the women. Compared with the HIV-1-seronegative women, the HIV-1-seropositive women showed more persistent HSV-2 genital shedding and higher HSV-2 DNA loads in eCVL. This study showed that eCVL is a reliable sampling method for detecting both viruses. Three measurement points (at weekly intervals in this study) seem to adequately allow detection of most HIV-1 or HSV-2 genital shedders. PMID- 16044020 TI - Atazanavir inhibits P-glycoprotein and multidrug resistance-associated protein efflux activity. PMID- 16044021 TI - The effect of statins on HIV rebound and blips. PMID- 16044022 TI - The cardiovascular continuum in Asia--a new paradigm for the metabolic syndrome. AB - Atherosclerosis, myocardial infarction, and heart failure are cardiovascular complications in a continuum that begins with risk factors such as hypertension, diabetes, and dyslipidemia. These particular cardiovascular risk factors commonly occur together in obese individuals as components of the metabolic syndrome. In Asia, there is a trend toward an increase in the prevalence of the metabolic syndrome and cardiovascular disease. Abdominal adiposity is arguably the key factor underlying the development of insulin resistance and the metabolic syndrome. It is now known that adipose tissues secrete adipokines, and in obese subjects, there is a chronic low-grade inflammation. The inflammation and the associated endothelial dysfunction are reversible in the early stages. The Asian diet is low in animal fat but high in carbohydrates. Recent studies suggest that low-carbohydrate diets are more effective than low fat diets in inducing weight loss, suggesting that excessive carbohydrate rather than fat is the cause of obesity. Strategies to combat cardiovascular disease should now focus on tackling the epidemic of obesity and developing innovative and effective lifestyle and pharmacological interventions. PMID- 16044023 TI - Labedipinedilol-C: a third-generation dihydropyridine-type calcium channel antagonist displaying K+ channel opening, NO-dependent and adrenergic antagonist activities. AB - Intravenous and oral labedipinedilol-C showed a dose-dependent long-lasting hypotension and a decrease of heart rate in normotensive and conscious spontaneously hypertensive rats (SHR). In isolated Wistar rat and guinea pig tissues, labedipinedilol-C competitively antagonized (-)isoproterenol-induced cardiac stimulation, tracheal relaxation, and phenylephrine-, CaCl2-, and high-K induced aorta contractions in a concentration-dependent manner. The estimated pA2 and pKCa values were 8.22+/-0.04 and 7.11+/-0.52, respectively. [H]CGP-12177 binding to ventricle and lung tissues as well as [H]prazosin and [H]nitrendipine binding to brain membranes were inhibited by labedipinedilol-C with Ki values of 2.86, 9.03, 0.39, and 0.05 muM, respectively. The vasorelaxant effects of labedipinedilol-C on phenylephrine (10 microM)-induced contractions were attenuated by removing endothelium, by pretreatment with soluble guanylyl cyclase (sGC) inhibitors ODQ (10 microM) and methylene blue (10 microM), a NOS inhibitor L-NAME (100 microM), a K channel blocker TEA (10 mM), a KATP channel blocker glibenclamide (1 microM), and Ca-dependent K channel blockers apamin (1 microM) and charybdotoxin (0.1 microM). In human umbilical vein endothelial cells (HUVECs), labedipinedilol-C increased NO release, which was significantly inhibited by L-NAME. The Western blot analysis on HUVECs indicated that labedipinedilol-C increased the expression of eNOS. These results indicate that hypotension effects of labedipinedilol-C result from alpha-adrenoceptor and Ca entry-blocking activities and release of NO or NO-related substance from vascular endothelium. The endothelium-independent relaxation of vascular smooth muscle is probably linked to K channel opening and alpha-adrenoceptor-blocking activities. PMID- 16044024 TI - Postischemic administration of CGX-1051, a peptide from cone snail venom, reduces infarct size in both rat and dog models of myocardial ischemia and reperfusion. AB - CGX-1051 is a synthetic version of a peptide originally isolated from the venom of cone snails. In the present studies, we tested the potential cardioprotective effect of CGX-1051 in a rat and dog model of myocardial ischemia/reperfusion. CGX 1051 was administered 5 minutes before reperfusion as intravenous bolus doses of 30, 100, and 300 microg/kg. Infarct size (IS) is reported as IS/area at risk (AAR). In the rat, the vehicle control group had an IS/AAR of 59.8+/-2.1%. Postischemic administration of CGX-1051 at doses of 30, 100, and 300 microg/kg resulted in an IS/AAR of 52.6+/-4.2%, 34.6+/-5.6% (P<0.05), and 40.8+/-5.2% (P<0.05), respectively. In the dog, the vehicle control group had an IS/AAR of 18.8+/-1.7%. Postischemic administration of CGX-1051 at doses of 30, 100, and 300 microg/kg resulted in an IS/AAR of 16.9+/-2.5%, 8.4+/-2.9% (P<0.05) and 9.9+/ 2.4% (P<0.05), respectively. These results demonstrate that administration of CGX 1051 at a clinically relevant time point results in a dose-dependent reduction in IS in both rats and dogs. PMID- 16044025 TI - Role of a responsive sympathetic nervous system in the chronic hypotensive effects of losartan in normal rats. AB - We have previously demonstrated the chronic hypotensive effects of the AT1 antagonist losartan in normotensive, salt-replete rats. We hypothesized that the chronic effects of losartan are mediated in part by blockade of the central sympathoexcitatory actions of angiotensin II. To test this hypothesis, we have used a novel approach to effectively "clamp" the sympathetic nervous system at a fixed level through chronic administration of the ganglionic blocking agent hexamethonium (15 mg/kg/h) and the alpha agonist phenylephrine (2.26 mg/kg/d). Two of 3 groups of rats [CON and CLAMP(NNa)] were placed on (0.1%) NaCl diets, whereas the third [CLAMP(LNa)] was placed on a low (0.002%)-sodium diet. Continuous measurements of mean arterial pressure (MAP) were made via radiotelemetry. After 9 days of hexamethonium plus phenylephrine treatment in CLAMP(NNa) and CLAMP(LNa) rats, baseline MAP was not different in all 3 groups of rats: CON (104+/-4 mm Hg), CLAMP(NNa) (104+/-4 mm Hg), and CLAMP(LNa) (106+/-2 mm Hg). After 5 days of subsequent losartan treatment, a change in MAP of only -7+/ 2 mm Hg was observed in CLAMP(NNa) rats compared with -22+/-2 mm Hg in CON and CLAMP(LNa) rats. These results do not support the hypothesis that the hypotensive actions of losartan are entirely dependent on a responsive sympathetic nervous system rats. PMID- 16044026 TI - Once-monthly administration of darbepoetin alfa for the treatment of patients with chronic heart failure and anemia: a pharmacokinetic and pharmacodynamic investigation. AB - In patients with chronic heart failure (CHF), anemia is associated with more severe symptoms and worse prognosis. Erythropoiesis-stimulating proteins (ESPs) increase hemoglobin and may be of therapeutic benefit. We investigated the pharmacokinetics and pharmacodynamics of the long-acting ESP, darbepoetin alfa, administered on 2 occasions 1 month apart to 30 healthy subjects and 33 patients with symptomatic CHF and anemia (hemoglobinspermidine>putrescine. In addition, spermine arrests or inhibits thrombin-, epinephrine-, arachidonate-, or ristocetin-induced platelet aggregation. Expression of platelet membrane glycoproteins IIb, IIIa, and IX is not reduced by polyamines. However, spermine inhibits the ADP- or thrombin induced activation of GP IIb/IIIa. It is concluded that the final step in aggregation, common to all agonists, ie, fibrinogen binding to GP IIb/IIIa, is inhibited by spermine through inhibition of the agonist-induced activation of GP IIb/IIIa that precedes fibrinogen-ligand binding. PMID- 16044035 TI - PET imaging of myocardial beta-adrenergic receptors with fluorocarazolol: lack of interference by endogenous catecholamines. AB - beta-Adrenergic receptor (beta-AR) concentration can be measured in vivo using positron emission tomography (PET) and the high-affinity antagonist [18F]-(S) fluorocarazolol {[18F]-(S)-FCZ}. However, the influence of endogenous catecholamines on the in vivo binding properties of [18F]-(S)-FCZ should be measured to aid in selection of the model used to estimate receptor concentration based on PET data. Herein we addressed the questions "What is the influence of endogenous catecholamines on the [18F]-(S)-FCZ binding in the heart?" and "In what range are the in vivo concentrations of endogenous beta-AR ligands?" In PET studies, 3 drug regimens were used to manipulate the levels of endogenous catecholamines. The time courses of myocardial concentration of [18F]-(S)-FCZ were compared before and after drug administration. In vitro binding assays and computer simulations were performed to complement the in vivo studies. Despite the large changes of endogenous catecholamines, no significant changes were observed in the [18F]-(S)-FCZ myocardial concentration. In vitro assays showed that (S)-FCZ has an affinity for beta-receptors that is 3900 and 9500 times higher than those of norepinephrine (NE) and epinephrine (EPI), respectively. Computer simulations support the hypothesis that the binding affinities relative to ligand concentrations in vivo are sufficient to explain the apparent lack of effect of endogenous catecholamines on [18F]-(S)-FCZ myocardial concentration. Increased levels of catecholamines in the physiological range do not affect the myocardial concentration of [18F]-(S)-FCZ as measured by PET. This lack of effect suggests that the myocardial concentration of NE at the synaptic sites cannot be higher than 300 nM. PMID- 16044036 TI - Pharmacological characterization of the novel dihydropyridine potassium channel opener, (9R)-9-(3-iodo-4-methylphenyl)-5,9-dihydro-3H-furo[3,4-b]pyrano[4,3 e]pyridine-1,8(4H,7H)-dione (A-325100), and the regulation of cardiovascular function in conscious and anesthetized beagle dogs. AB - The pharmacological profile of the novel dihydropyridine K channel opener (KCO), (9R)-9-(3-iodo-4-methylphenyl)-5,9-dihydro-3H-furo[3,4-b]pyrano[4,3-e]pyridine 1,8(4H,7H)-dione (A-325100), is described in numerous in vitro assays. Furthermore, the cardiovascular effects of A-325100 are characterized in both the anesthetized and conscious dog. In vitro, A-325100 selectively activated KATP currents and potently relaxed vascular smooth muscle (IC50 between 7.69x10 M and 7.78x10 M), an effect that was abolished by glyburide. Moreover, A-325100 did not interact with L-type Ca2+ channels at concentrations up to 30 microM. In anesthetized dogs A-325100 produced a dose-dependent reduction in systemic vascular resistance and mean arterial pressure concomitant with dose-dependent increases in dP/dtmax and heart rate. In conscious telemetry-instrumented dogs oral administration of A-325100 produced a similar response profile, including dose-dependent reductions in MAP and increases in heart rate and dP/dtmax. When concentration-dependent changes in MAP, heart rate, and dP/dtmax were compared relative to circulating plasma concentrations, A-325100 produced similar effects in both the anesthetized and conscious dog. In conclusion, the present study provides the first pharmacological description of the novel and selective tricyclic dihydropyridine KCO, A-325100. When studied in vivo, A-325100 produced similar concentration-dependent cardiovascular effects in both models consistent with its mode of action and independent of route of administration. Thus, these data demonstrate that the hemodynamic effects of vasoactive compounds, such as KCOs, can be effectively profiled in both the conscious and anesthetized dog. PMID- 16044037 TI - Essential elements of evidenced-based endodontics: steps involved in conducting clinical research. AB - Endodontists have the opportunity to apply relevant research findings to the care of their patients using the principles and methods of evidence-based treatment. Finding evidence begins with a specific, well-built clinical question. Once a specific question is framed, the validity and relevance of the evidence need to be appraised. The best levels of evidence can then be used to inform decisions regarding care. The purpose of this paper is to discuss the history of evidence based treatment and to clarify the process of conducting a systematic review. The various types of research designs appropriate for answering clinical questions most commonly encountered in dental practice, including a description of the strengths and weaknesses of each, are also presented. Finally, the implications of evidence-based research on endodontics and future research are outlined. PMID- 16044038 TI - Morphological measurements of anatomic landmarks in pulp chambers of human maxillary furcated bicuspids. AB - The aim of this in vitro study was to measure key morphological features of pulp chambers from furcated maxillary bicuspid teeth. There were 107 random human maxillary bicuspid teeth used. Each bicuspid was radiographed using the Trophy RVG digital imaging system and a Belmont Acuray X-ray at 70 kVp. Measurements were made using the Digipan measuring mode of the Trophy system. Results were mean (mm): pulp chamber floor to furcation: 1.85 +/- 0.85; pulp chamber ceiling to furcation: 4.61 +/- 1.04; cusp to furcation: 11.55 +/- 1.12; cusp to pulp chamber ceiling: 6.94 +/- 0.70; pulp chamber height: 2.76 +/- 0.97. The measurements showing the lowest percentage variance were: cusp to furcation (9.70%) and cusp to pulp chamber ceiling (10.09%). The only measurement that was statistically the same across maxillary molars, mandibular molars and bicuspids was measurement "B," pulp chamber ceiling to furcation. The critical distance from cusp tip to pulp chamber ceiling in bicuspids was approximately 7.00 mm. PMID- 16044039 TI - Immunolocalization of receptor activator of NF kappa B ligand in rat periapical lesions. AB - The purpose of this study is to observe the immunohistochemical localization of Receptor Activator of NF Kappa B Ligand (RANKL) in the rat periapical lesions. Periapical lesions were induced in Sprague-Dawley (SD) rats by occlusal exposure of the pulp of their mandibular first molars. Animals were sacrificed randomly at 0, 7, 14, 21, and 28 days after the exposure and their mandibles were removed, demineralized, and embedded in paraffin. Frontal sections in the region of the first molar were prepared for enzyme histochemistry and immunohistochemistry. Tartrate-resistant acid phosphatase (TRAP)-positive cells and RANKL-positive cells could be observed on day 7. From day 7 to day 14, increasing amounts of inflammatory infiltration and alveolar bone resorption were observed in the periapical region. The expression and number of TRAP-positive cells and RANKL positive cells were climaxed on day 14. In the 21- and 28-day samples, the expression of RANKL decreased and fewer TRAP-positive cells could be observed. These findings demonstrated that RANKL could be observed, which may be associated with the pathogenesis of periapical lesions. PMID- 16044040 TI - Effects of pre-emptive morphine, ibuprofen or local anesthetic on fos expression in the spinal trigeminal nucleus following tooth pulp exposure in the rat. AB - In the present study, we used Fos expression as an index of nociceptive input to the spinal trigeminal nucleus after exposure of the coronal pulp tissue of maxillary right first molars and examined the effects of pretreatment with an opioid, a nonsteroidal anti-inflammatory drug or a local anesthetic before pulp exposure. Exposure of the tooth pulp produced a significant increase in Fos-like immunoreactivity in the superficial laminae of subnucleus caudalis; pretreatment with a control infiltration injection of saline directly above the maxillary molar 30 min before pulp exposure had no effect on Fos expression. Pretreatment with morphine 30 min before pulp exposure dose-dependently (2.5, 5, and 10 mg/kg subcutaneously) reduced Fos expression in subnucleus caudalis whereas pretreatment with ibuprofen (10-100 mg/kg subcutaneously) did not significantly affect Fos expression. Local anesthetic pretreatment was effective in reducing Fos expression only for the long acting bupivacaine; lidocaine without and with epinephrine (1:100,000) failed to significantly affect Fos expression. These results suggest that pre-emptive opioid treatment can decrease postoperative central nervous system changes associated with tooth pulp injury, and therefore, may decrease postoperative pain. Given the effects of local anesthetic on Fos expression, a combination of long acting local anesthetic with pre-emptive opioid would likely be most efficacious in decreasing postoperative dental pain. PMID- 16044041 TI - Geometric factors affecting dentin bonding in root canals: a theoretical modeling approach. AB - Cavity configuration factor (C-factor) is the ratio of the bonded surface area in a cavity to the unbonded surface area. In a box-like class I cavity, there may be five times more bonded surface area than the unbonded surface area. During polymerization, the volume of monomers is reduced, which creates sufficient shrinkage stresses to debond the material from dentin, thereby decreasing retention and increasing leakage. The important variables influencing bonding adhesive root-filling materials to canals was examined using a truncated inverted cone model. C-factors in bonded root canals exhibit a negative correlation with sealer thickness. For a 20 mm-long canal prepared with a size 25 file, calculated C-factors ranged from 46 to 23,461 with decreasing sealer thickness (500-1 microm), compared to a C-factor of 32 when the canal was filled only with sealer. As the thickness of the adhesive is reduced, the volummetric shrinkage is reduced, which results in a reduction in shrinkage stress (S-factor). C-factors above 954 calculated with sealer thickness smaller than 25 microm are partially compensated by increases in bonding area and decreases in shrinkage volume. However, the interaction of these two geometrically related factors (C- and S factors) predicts that bonding of adhesive root-filling materials to root canals is highly unfavorable when compared with indirect intracoronal restorations with a similar resin film thickness. PMID- 16044042 TI - Reducing surface tension in endodontic chelator solutions has no effect on their ability to remove calcium from instrumented root canals. AB - The aim of this study was to evaluate the effect of reducing surface tension in endodontic chelator solutions on their ability to remove calcium from instrumented root canals. Aqueous solutions containing 15.5% EDTA, 10% citric acid, or 18% 1- hydroxyethylidene-1, 1-bisphosphonate (HEBP) were prepared with and without 1% (wt/wt) polysorbate (Tween) 80 and 9% propylene glycol. Surface tension in these solutions was measured using the Wilhelmy method. Sixty-four extracted, single-rooted human teeth of similar length were instrumented and irrigated with a 1% sodium hypochlorite solution and then randomly assigned (n = 8 per group) to receive a final one-minute rinse with 5 ml of test solutions, water, or the pure aqueous Tween/propylene glycol solution. Calcium concentration in eluates was measured using atomic absorption spectrometry. Incorporation of wetting agents resulted in a reduction of surface tension values by approximately 50% in all tested solutions. However, none of the solutions with reduced surface tension chelated more calcium from canals than their pure counterparts (p > 0.05). PMID- 16044043 TI - Susceptibility of a polycaprolactone-based root canal filling material to degradation. I. Alkaline hydrolysis. AB - Polycaprolactone, a thermoplastic aliphatic polyester, is reportedly susceptible to both alkaline and enzymatic hydrolyzes. This screening study examined the susceptibility of Resilon, a polycaprolactone-based root filling composite, to alkaline hydrolysis. There were 15-mm diameter disks of Resilon and Obtura gutta percha prepared by compressive molding and immersed in 20% sodium ethoxide for 20 or 60 min. Control disks were immersed in ethanol for 60 min. These disks were examined using field-emission scanning electron microscopy and energy dispersive X-ray analysis. For Resilon, the surface resinous component was hydrolyzed after 20 min of sodium ethoxide immersion, exposing the spherulitic polymer structure and subsurface glass and bismuth oxychloride fillers. More severe erosion occurred after 60 min of sodium ethoxide treatment. Gutta-percha was unaffected after immersion in sodium ethoxide. As Resilon is susceptible to alkaline hydrolysis, it is possible that enzymatic hydrolysis may occur. Biodegradation of Resilon by bacterial/salivary enzymes and endodontically relevant bacteria warrants further investigation. PMID- 16044044 TI - Effect of root canal filling/sealer systems on apical endotoxin penetration: a coronal leakage evaluation. AB - Endotoxin, elaborated by gram-negative organisms, is an important factor in apical periodontitis. The objective of this study was to evaluate the magnitude of endotoxin penetration through root canal treated teeth using a dual chamber model system. Forty-four maxillary anterior teeth were prepared endodontically and canals filled either by lateral condensation or a warm thermoplasticized technique in combination with either Roth's 801 or AH 26 sealer. Teeth were suspended in the model system with a mixed anaerobic bacterial suspension in the upper chamber and HBSS in the lower chamber. The QCL-1000 LAL assay was used to measure endotoxin at 0, 1, 7, 14, and 21 days. Response feature analysis using trapezoidal area under the curve was performed; the four treatment groups were compared using nonparametric methods. Groups differed (p = 0.028), with thermoplasticized root canal filling/Roth's 801 sealer permitting the least apical endotoxin penetration. Results suggest that Roth's 801 sealer may have a role in inhibiting endotoxin penetration. PMID- 16044045 TI - Biocompatibility in vitro tests of mineral trioxide aggregate and regular and white Portland cements. AB - Mineral trioxide aggregate (MTA) and Portland cement are being used in dentistry as root end-filling materials. However, biocompatibility data concerning genotoxicity and cytotoxicity are needed for complete risk assessment of these compounds. In the present study, genotoxic and cytotoxic effects of MTA and Portland cements were evaluated in vitro using the alkaline single cell gel (comet) assay and trypan blue exclusion test, respectively, on mouse lymphoma cells. The results demonstrated that the single cell gel (comet) assay failed to detect DNA damage after a treatment of cells by MTA and Portland cements for concentrations up to 1000 microg/ml. Similarly, results showed that none of the compounds tested were cytotoxic. Taken together, these results seem to indicate that MTA and Portland cements are not genotoxins and do not induce cellular death. PMID- 16044046 TI - The contribution of friction to the dislocation resistance of bonded fiber posts. AB - This study tested the null hypothesis that the use of dentin adhesives produces no improvement on the fixation of fiber posts with resin cements in endodontically treated teeth. Post spaces were prepared in 36 single-rooted root filled teeth. Silanized glass fiber posts were cemented to the post spaces using a self-etch (ED primer/Panavia 21) and a total etch resin cement (Excite DSC/Variolink II), with or without the accompanying dentin adhesives. Fixation strengths and interfacial ultrastructure were evaluated using a "thin slice" push out test and transmission electron microscopy. For both resin cements, the fixation strengths obtained from specimens luted with resin cement only did not differ significantly from those in which the intraradicular dentin was first bonded with a dentin adhesive. In the presence of incomplete smear layer removal and interfacial gaps, the dislocation resistance of bonded fiber posts was contributed largely by sliding friction. PMID- 16044047 TI - An in vitro evaluation of the cytotoxicity of various endodontic irrigants on human gingival fibroblasts. AB - The purpose of this study was to measure the cytotoxicity of six endodontic irrigants on cultured gingival fibroblasts using the CyQuant assay. Human gingival fibroblasts were grown in Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum at 37 degrees C and 5% CO(2). At confluence, cells were split, plated in 96-well plates and incubated for 24-h to allow attachment. The following irrigants were tested at various concentrations: Sodium hypochlorite (NaOCl); iodine potassium-iodide (IKI); Betadine scrub (BS); calcium hydroxide [Ca(OH)2]; chlorine dioxide (SCD) and DMEM (positive control). Experimental groups were compared by the logarithmic difference between the clinical and LD50 concentrations of a particular irrigant. The results showed that IKI and Ca(OH)2 were significantly less cytotoxic than SCD, NaOCl, and BS. In conclusion, IKI and Ca(OH)2 are well tolerated by human gingival fibroblasts. PMID- 16044048 TI - A novel method for creating endodontic access preparations through all-ceramic restorations: air abrasion and its effect relative to diamond and carbide bur use. AB - Access through porcelain restorations is a technically delicate and stressful procedure. Although this is a common dilemma in endodontics, little research has explored alternatives in cutting through porcelain. The purpose of this study was to compare the use of a carbide bur plus water, diamond bur plus water, and air abrasion to access through porcelain. All-ceramic samples were accessed using the different techniques. Samples were evaluated using two transillumination methods, white light, and fluorescent liquid penetrant described by the American Society for Testing and Materials. Edge chipping, microcracking, and catastrophic fracture of porcelain caused by the techniques were statistically compared. Fluorescent liquid penetrant was a more sensitive method for microcrack detection. There were significant differences between the preparation techniques. Air abrasion was significantly less destructive, and caused no catastrophic fractures, edge chipping or microcracks. Preparation by air abrasion took longer to complete. PMID- 16044049 TI - Sealer penetration and apical microleakage in smear-free dentin after a final rinse with either 70% isopropyl alcohol or Peridex. AB - Tensioactive agents may alter dentinal wettability allowing increased sealer penetration into dentinal tubules. This may improve the apical seal and better entomb remaining bacteria. The purpose of this study was to determine if a final rinse with either 70% isopropyl alcohol or Peridex in instrumented, smear-free canals would affect apical microleakage or Roth's 801 sealer penetration into dentinal tubules. Sixty root segments were instrumented followed by smear layer removal. The segments were randomly divided into three groups receiving a final rinse with 6% sodium hypochlorite (NaOCl), 70% isopropyl alcohol or Peridex. Obturation was completed using lateral compaction and Roth's 801 sealer pigmented with carbon black. Microleakage was measured by fluid filtration. Sealer penetration was measured using light microscopy. No significant differences were found between groups for microleakage or sealer penetration. Although a final rinse with either 70% isopropyl alcohol or Peridex did not increase sealer penetration, neither significantly affected apical microleakage compared to NaOCl. PMID- 16044051 TI - Can advanced practice clinicians perform loop electrosurgical excision procedures and cryotherapy? PMID- 16044053 TI - Managing abnormal cytology results in pregnancy. PMID- 16044054 TI - Characterization of papillary squamotransitional cell carcinoma of the cervix. AB - OBJECTIVE: Does papillary squamotransitional cell carcinoma (PSTCC) behave differently from conventional squamous cell carcinoma of the cervix and does PSTCC have true transitional cell differentiation? MATERIALS AND METHODS: Twenty cases of PSTCC were identified from archival files. Clinical data were compiled. Immunoperoxidase stains for uroplakin III, p63 and p16 were performed on available tissue blocks. RESULTS: Patients ranged in age from 27 to 85 years. Twelve patients were FIGO Stage I, 4 were Stage II, and 2 were Stage III. Thirteen patients had clinical follow-up ranging from 5 to 132 months. Three patients subsequently had more extensive disease than initial clinical staging indicated. Nine patients had no tumor progression, three had local recurrence and one had metastatic disease. Eight cases were strongly immunoreactive for p63 and p16 and 14 were negative for uroplakin III. CONCLUSIONS: PSTCC lacks true transitional cell differentiation and probably shares similar clinicopathologic features with conventional cervical squamous cell carcinoma. PMID- 16044055 TI - Type-specific HPV testing as a predictor of high-grade squamous intraepithelial lesion outcome after cytologic abnormalities. AB - OBJECTIVE: Human papillomavirus (HPV) risk assignment influences management after a cytologic diagnosis of atypical squamous cells of undetermined significance (ASCUS) or low-grade squamous intraepithelial lesion (LSIL). This study addressed whether type-specific HPV testing predicts risk of biopsy outcome of high-grade cervical intraepithelial neoplasia (CIN 2,3). MATERIALS AND METHODS: A total of 162 ASCUS or LSIL diagnoses with colposcopic follow-up were evaluated and placed in 3 groups: Analysis 1: (high-risk HPVs including types 53 and 66; Analysis 2 (high-risk HPVs excluding types 53 and 66); and Analysis 3 (high-risk HPVs including type 53 and excluding type 66). RESULTS: CIN 2,3 biopsy results followed low-risk HPVs in 0%, 3%, and 0% of scenarios 1, 2, and 3, respectively; 21%, 40%, and 27% of smears were classified as low risk, respectively. Of HPV 53 infections, 13.6% had CIN 2,3 biopsy outcomes. CONCLUSIONS: Type-specific HPV testing accurately classifies a group of HPV-positive LSIL/ASCUS cases at low risk for CIN 2,3 at the first follow-up visit. Classifying HPV 53 as low-risk increases slightly the proportion of HSIL outcomes in the low-risk group, but may not increase cancer risk. HPV 53 merits designation as a high-risk HPV based only [corrected] on the proportion of CIN 2,3 in follow-up biopsy. PMID- 16044056 TI - Costs of colposcopy services and their impact on the incidence and mortality rate of cervical cancer in Canada. AB - OBJECTIVE: Organized cervical cancer screening services consisting of conventional Papanicolaou cervical smears, colposcopy, and related treatment modalities are readily available in all provinces. The purpose of this report was to study the impact of colposcopy usage and costs on cervical cancer incidence and mortality rates in several Canadian provinces. Knowledge of such information is essential before newer technology such as liquid-based cytology and human papillomavirus testing is introduced or replaces the traditional systems used. MATERIALS AND METHODS: The Ministries of Health of five provinces were contacted and asked to furnish information on the number of colposcopic services and fee for-service costs for these and for cryosurgery, carbon dioxide laser vaporization, loop electrosurgical excisions, and cold-knife conizations for the year 2000. Canadian Cancer Society estimates of incidence and mortality rates for cervical cancer were also obtained. RESULTS: All provinces had similar incidence and mortality rates for cervical cancer; however, the number of colposcopic services on a per-capita basis varied substantially, with Manitoba and Ontario having rates that were approximately two or three times higher. Fee-for-service payments for colposcopy were similar in the Provinces studied but unit costs for surgical treatment services were highest in Ontario and British Columbia. CONCLUSIONS: Although both the incidence and mortality rates for cervical cancer in Canada fell dramatically after the Walton Report in 1976, these rates have plateaued over the past decade despite widespread availability of colposcopy and related ambulatory treatment services. Higher rates of colposcopy usage do not seem to result in lower incidence rates for this disease. Unit costs for colposcopy are similar among the provinces reviewed, but substantial difference exists for certain treatment services. Additional studies are recommended before the widespread introduction or replacement of existing methods with newer, more costly techniques. PMID- 16044057 TI - Successful treatment of refractory recurrent vaginal candidiasis with cetirizine plus fluconazole. AB - OBJECTIVE: To determine the prevalence of allergy in patients with recurrent vaginal candidiasis (RVC) and the efficacy of fluconazole in combination with an antihistamine drug in RVC patients who do not respond to fluconazole alone. MATERIALS AND METHODS: Participants in the study included 24 women with RVC. All subjects filled out an International Study of Asthma and Allergies in Childhood questionnaire and 23 performed skin prick tests for immediate type 1 hypersensitivity as part of atopy assessment. All subjects were treated with fluconazole 150 mg weekly and were followed every 2 months for 6 months. Those who did not respond to fluconazole alone received cetirizine 10 mg daily in addition to the antifungal medication and were followed up for 6 months. RESULTS: The majority of women had a history of allergy (60%) and 10 of them (43%) had at least 1 positive result of a skin prick test to aeroallergens. Nineteen patients (79%) responded to fluconazole alone. The 5 patients who remained with symptoms had a medical history of allergy and obtained remission when cetirizine was associated with fluconazole. CONCLUSIONS: Patients with RVC refractory to standard antifungal therapy and a history of allergy may respond to the combination of antihistamine and antifungal therapy. PMID- 16044058 TI - A double-blind, randomized, placebo-controlled trial of prilocaine and felypressin (Citanest and Octapressin) for the relief of pain associated with cervical biopsy and treatment with the Semm coagulator. AB - OBJECTIVE: To quantify the anticipated and actual pain experienced in association with preliminary cervical punch biopsies and subsequent ablative treatment with the Semm coagulator, and to test the hypothesis that the intracervical injection of prilocaine with felypressin reduces the intensity of the pain experienced. MATERIALS AND METHODS: One hundred consecutive women referred with abnormal cervical smears for colposcopic assessment and considered suitable for treatment with the Semm coagulator were recruited to a double-blind, randomized, prospective, placebo-controlled trial conducted in a colposcopy clinic in a university teaching hospital. Personal particulars were taken and anticipated pain scored. The patients were injected with randomized externally identical vials of prilocaine and felypressin (Citanest and Octapressin) or placebo. After biopsy and treatment, patients scored their actual pain experienced. Pain scores were compared as the main outcome measure. Relative risks with 95% CIs were calculated and compared using the CI Analysis computer programme (Professor Martin J Gardner and the British Medical Journal Version 1.1, copyright 1991). RESULTS: Anticipated pain was greater than the actual pain experienced in both groups. Women receiving the local anesthesia experienced a significantly greater reduction in pain (p < .05) with only 4.3% and 6.7% experiencing moderate pain during biopsy and treatment, respectively. The active drug abolished severe pain. In the placebo group, 44.7% felt mild pain at the most. CONCLUSIONS: Intracervical injection of prilocaine and felypressin reduces the intensity of pain experienced in women undergoing cervical biopsy and treatment with the Semm coagulator. Its use is commendable but is not absolutely necessary in all cases. PMID- 16044059 TI - Topical misoprostol therapy for plasma cell vulvitis: a case series. AB - OBJECTIVE: To present the first reported case series of plasma cell vulvitis successfully treated with topical misoprostol. MATERIALS AND METHODS: Three women with vulvar pain and lesions secondary to plasma cell vulvitis refractory to conventional therapy were treated with topical misoprostol compounded in white petrolatum. RESULTS: All three patients had resolution of pain and lesions within 6 weeks of starting topical misoprostol. All patients tolerated the 0.01% concentration; however, local side effects were seen with the higher 0.02% concentration. CONCLUSIONS: Plasma cell vulvitis is a rare and painful vulvar disorder that is often refractory to therapy. All three patients in this case series were successfully treated with topical misoprostol. The immunosuppressive effects of misoprostol are hypothesized to be the mechanisms of action of this novel therapy for plasma cell vulvitis. PMID- 16044060 TI - Home study course: summer 2005. AB - OBJECTIVE: The Home Study Course is intended for the practicing colposcopist or practitioner who is seeking to develop or enhance his or her colposcopic skills. The goal of the course is to present colposcopic cases that are unusual or instructive in terms of appearance, presentation, or management, or that demonstrate new and important knowledge in the area of colposcopy or pathology. Participants may benefit from reading and studying the material or from testing their knowledge by answering the questions. ACCME ACCREDITATION: The American Society for Colposcopy and Cervical Pathology (ASCCP) is accredited by the Accreditation Council for Continuing Medical Education (ACCME) to provide continuing medical education for physicians. The ASCCP designates this continuing medical education activity for 1-hour Category I credit of the Physician's Recognition Award of the American Medical Association and 1-hour Category I credit of the ASCCP's Program for Continuing Professional Development. Credit is available for those who choose to apply. The Home Study Course is planned and produced in accordance with the ACCME's Essential Areas and Elements. PMID- 16044061 TI - Clinical question: ask the experts. PMID- 16044063 TI - A randomized clinical trial of vision therapy/orthoptics versus pencil pushups for the treatment of convergence insufficiency in young adults. AB - PURPOSE: : The purpose of this article is to compare vision therapy/orthoptics, pencil pushups, and placebo vision therapy/orthoptics as treatments for symptomatic convergence insufficiency in adults 19 to 30 years of age. METHODS: : In a randomized, multicenter clinical trial, 46 adults 19 to 30 years of age with symptomatic convergence insufficiency were randomly assigned to receive 12 weeks of office-based vision therapy/orthoptics, office-based placebo vision therapy/orthoptics, or home-based pencil pushups. The primary outcome measure was the symptom score on the Convergence Insufficiency Symptom Survey. Secondary outcome measures were the near point of convergence and positive fusional vergence at near. RESULTS: : Only patients in the vision therapy/orthoptics group demonstrated statistically and clinically significant changes in the near point of convergence (12.8 cm to 5.3 cm, p = 0.002) and positive fusional vergence at near (11.3Delta to 29.7Delta, p = 0.001). Patients in all three treatment arms demonstrated statistically significant improvement in symptoms with 42% in office based vision therapy/orthoptics, 31% in office-based placebo vision therapy/orthoptics, and 20% in home-based pencil pushups achieving a score <21 (our predetermined criteria for elimination of symptoms) at the 12-week visit. DISCUSSION: : In this study, vision therapy/orthoptics was the only treatment that produced clinically significant improvements in the near point of convergence and positive fusional vergence. However, over half of the patients in this group (58%) were still symptomatic at the end of treatment, although their symptoms were significantly reduced. All three groups demonstrated statistically significant changes in symptoms with 42% in office-based vision therapy/orthoptics, 31% in office-based placebo vision therapy/orthoptics, and 20% in home-based pencil push-ups meeting our criteria for elimination of symptoms. PMID- 16044064 TI - The research-based case against "drug store" reading glasses. PMID- 16044068 TI - The development of a symptom questionnaire for assessing virtual reality viewing using a head-mounted display. PMID- 16044069 TI - Bilateral, persistent serous macular detachments with Waldenstrom's macroglobulinemia. AB - PURPOSE: Waldenstrom's macroglobulinemia is a rare, malignant lymphoplasmacytic disorder characterized by the monoclonal proliferation of immunoglobulin type M (IgM) producing B-lymphocytes. Ocular manifestations of Waldenstrom's macroglobulinemia have been described in association with the conjunctiva, cornea, uvea, retina, and periocular adnexa. Only rarely have macular findings, particularly serous macular detachments, been described in the presence of monoclonal hypergammaglobulinemias. The majority of reports to date have documented resolution of these serous macular detachments after treatment with blood plasmapheresis. This report presents a case of bilateral, persistent serous macular detachments in the presence of Waldenstrom's macroglobulinemia. CASE REPORT: A 53-year-old black man presented with gradual bilateral reduction in visual acuity over the last 2 months. He revealed having been diagnosed with Waldenstrom's macroglobulinemia approximately 1 month before presentation. He also reported having undergone multiple plasmapheresis treatments in conjunction with systemic chemotherapy over the same period. Funduscopic and fluorescein angiographic examination revealed bilateral, serous macular detachments in the presence of mild venous stasis retinopathy secondary to serum hyperviscosity. Subsequent funduscopic evaluations and serial optical coherence tomography readings confirmed the persistence of the serous detachments despite multiple plasmapheresis treatments. CONCLUSIONS: Although the funduscopic manifestations of excess serum immunoglobulins are typically the sequelae of circulatory stasis, the serum hyperviscosity induced in Waldenstrom's macroglobulinemia has been shown on rare occasions to induce serous macular detachments. Physical elimination of the excess macroglobulins through plasmapheresis is quite effective in resolving the neurosensory detachments induced by Waldenstrom's macroglobulinemia. Only a fraction of neurosensory detachments resulting from Waldenstrom's macroglobulinemia persist despite multiple plasmapheresis treatments. Novel angiographic evidence of retinal pigment epithelium atrophy beneath the area corresponding to the serous detachment provides a plausible explanation for unresponsive nature of this presentation. Identification of this rare variant of the classic angiographically silent macula known to occur with Waldenstrom's macroglobulinemia can aid the clinician in determining a patient's potential response to plasmapheresis and their final visual prognosis. PMID- 16044070 TI - Madarosis and facial alopecia presumed secondary to botulinum a toxin injections. AB - PURPOSE: : Botulinum neurotoxins are considered relatively safe therapy for treatment of inappropriate muscle spasms. As seen in this report, undesirable consequences may result. METHODS: : The author conducted a clinical interview and examination of the patient. RESULTS: : Unilateral left-sided madarosis and facial alopecia were observed in a patient undergoing botulinum A toxin injections for the treatment of left oromandibular dystonia. The patient had started treatment 18 months previously. He received injections every 3 months in the left masseter and left temporalis muscles. He first noted loss of his left sideburn 8 months into treatment. After 10 months of treatment, he noted that he no longer needed to shave on the left side of his face. Eighteen months after receiving his first botulinum toxin injection, madarosis of the temporal aspect of the left lower lid was observed. CONCLUSIONS: : This is the first case report documenting an idiosyncratic unilateral madarosis and facial alopecia as adverse side effects presumed secondary to botulinum A toxin injections. PMID- 16044071 TI - The effect of two novel lubricant eye drops on tear film lipid layer thickness in subjects with dry eye symptoms. AB - PURPOSE: Because the lipid layer of the tear film is recognized as a critical component in dry eye disease, this study was designed to determine if a single eye drop of either Soothe or Systane produces a significant increase in lipid layer thickness (LLT) for subjects reporting symptoms indicative of dry eyes. METHODS: A double-blind, internally paired study was performed. A custom-built lipid layer interferometer, enabling characterization of lipid layer interference patterns, was used to quantify baseline LLT (OU) of eligible subjects. Inclusion criteria included: 1) presence of dry eye symptoms and 2) baseline LLT < or =75 nm. Subjects (n = 40) received a single eye drop of Soothe in one eye and a single eye drop of Systane in the contralateral eye. After the instillation of each test drop, LLT was reanalyzed for all subjects. RESULTS: The mean +/- standard error baseline LLT pre-eye drop instillation was 60.0 +/- 1.8 nm for eyes treated with Soothe and 61.5 +/- 1.8 nm for eyes treated with Systane. These means were not significantly different (p > 0.5). The mean LLT for eyes treated with Soothe increased to 124.4 +/- 4.9 nm (p < 0.0001). The mean LLT for eyes treated with Systane increased to 71.3 +/- 2.6 nm (p < 0.0001). The LLT increase from Soothe was significantly greater than that from Systane (p < 0.0001). CONCLUSIONS: In subjects with symptoms indicative of dry eye states and LLT < or =75 nm, one eye drop of Soothe more than doubled LLT, a 107% mean increase, whereas Systane increased LLT by 16%. PMID- 16044072 TI - The effect of common reductions in letter size and contrast on accommodation responses in young adult myopes and emmetropes. AB - PURPOSE: Accommodation errors during reading and the subsequent near work-induced transient myopia (NITM) that occurs have been implicated in the development and progression of myopia. This study investigated the effects of two letter variables, size and contrast, on accommodation accuracy during the near task and on NITM and its subsequent decay. These were varied so as to mimic what might occur when students photocopy and reduce reading materials. METHODS: Based on their refractive errors, young adult subjects (18-25 years) were classified into three groups: emmetropes (n = 19), stable myopes (n = 17), and progressing myopes (n = 17). Three print sizes (N4, N6, and N8) and two print contrasts (90% and 60%) were used to give six different reading targets. Targets were presented in random order at 25 cm (4 D demand) and the text read for comprehension for 3 minutes. For each target, accommodation accuracy and NITM and its decay were measured using the free space Shin-Nippon SRW-5000 autorefractor. RESULTS: When data for all subjects were pooled, there was a significant effect of letter size (p = 0.030) but not contrast (p = 0.898) on accommodation accuracy; however, differences were small and unlikely to be clinically relevant. NITM (p = 0.033) and its decay (p = 0.012) also varied with letter size. NITM was greater and decay longer for larger letters. We found no effect of refractive error group on accommodation accuracy. In contrast, there was a significant difference in the magnitude of NITM and its decay for emmetropic and myopic subjects (although no effect of progression status); myopes had larger NITM values and longer decay times to baseline than emmetropes (NITM myopes: 0.37 +/- 0.14D vs. emmetropes: 0.19 +/- 0.17 D, p = 0.005; decay time myopes: 15.12 +/- 6.58 seconds vs. emmetropes 7.10 +/- 4.82 seconds, p = 0.0045). The differences in NITM and its decay between the two refractive groups were of similar magnitude for all six combinations of letter size and contrast. CONCLUSIONS: Our data do not support the suggestion that common reductions in letter size or contrast of reading material (as might occur for photocopied reading materials) cause greater accommodation inaccuracy and greater near work-induced adaptation effects that would exacerbate myopia development in young adults. PMID- 16044073 TI - Factors related to vision care in an older adult cohort. AB - PURPOSE: This study provides cross-sectional data on eye care utilization in a community-based adult population. METHODS: Data are from a questionnaire administered during the 7-year follow-up of the Epidemiology of Hearing Loss Study in 2000 to 2002. Participants in the population-based Beaver Dam Eye Study were eligible for the Epidemiology of Hearing Loss Study, which began in 1997. The primary outcome was self-reported vision testing within the past year. RESULTS: Subject ages ranged from 55 to 99 years (n = 2433), and 60.4% were female. Fifty-three percent of subjects reported they had their vision tested in the past year. Diabetes was self-reported by 11.5% of subjects, and 70.9% of diabetic participants had their vision tested in the past year. A current hospitalization or health insurance plan was reported by 98.6% of subjects. In multivariate analyses, self-reported factors significantly associated with having a vision test in the past year were female gender (odds ratio [OR] = 1.27; 95% confidence interval [CI] 1.06-1.52), current use of refractive correction for distance (OR = 1.98; 95% CI 1.56-2.52), glaucoma (OR = 3.52; 95% CI 2.37-5.24), cataract surgery (OR = 1.57; 95% CI 1.21-2.03), age-related macular degeneration (ARMD) (OR = 1.74; 95% CI 1.22-2.47), diabetes (OR = 2.46; 95% CI 1.83-3.31), visiting a primary care practitioner for any reason in the past year (OR = 1.72; 95% CI 1.32-2.25), having a hearing test in the past year (OR = 1.79; 95% CI 1.40 2.28), and the cessation of driving because of poor vision (OR = 1.64; 95% CI 1.16-2.52). In participants 65 years of age or older, having private insurance was associated with increased odds (OR = 3.39, 95% CI 1.82-6.31) of vision testing in the past year. CONCLUSION: This study suggests that chronic ocular conditions, diabetes, health insurance beyond government entitlements, and the use of other healthcare services are associated with increased eye care utilization. PMID- 16044074 TI - Thirty years in an urban low vision clinic: changes in prescribing habits of low vision practitioners. AB - PURPOSES: The purposes of this study were to determine the types of low vision devices (LVDs) prescribed by clinicians in a large optometrist led hospital low vision clinic and to investigate changes in these prescribing habits between 1973 and 2003. METHODS: Data were collected from departmental records for all patients attending the low vision clinic at Moorfields Eye Hospital in London, U.K., during the month of June in 1973, 1978, 1983, 1988, 1993, 1998, and 2003. The primary outcome measure was the type or types of LVDs prescribed to patients attending the clinic for the first time. Data were collected on the age, sex, and diagnosis of all patients attending the clinic. RESULTS: One thousand four hundred thirty-eight patient visits were identified, of which 563 (39%) were first attendances. The median age of the patients examined did not change significantly over this time. Eighty-four percent of the patients attending the clinic for the first time were prescribed at least one LVD. The devices most frequently prescribed to new patients were nonilluminated hand magnifiers, illuminated hand magnifiers, and illuminated stand magnifiers. There was a linear increase in the proportion of hand magnifiers prescribed between 1973 and 2003 (r = 0.62, p < 0.05) and a corresponding decrease in the number of near spectacle mounted telescopes prescribed (r = 0.70, p < 0.05). CONCLUSIONS: Between 1973 and 2003, the low vision clinic at Moorfields Eye Hospital provided nearly 100,000 low vision consultations. The demographics of patients attending the clinic remained remarkably constant over this time. The number of spectacle-mounted devices prescribed to new patients fell between 1973 and 2003. It is thought this is in part the result of the increased availability of electronic magnification devices and the development of illuminated hand magnifiers. The development of the acrylic bright field magnifier and LED-illuminated LVDs have had particular clinical impact. PMID- 16044075 TI - Higher-order wavefront aberrations in retinitis pigmentosa. AB - PURPOSE: The purpose of this study was to characterize higher-order wavefront aberrations associated with posterior subcapsular (PSC) cataracts in patients with retinitis pigmentosa (RP). METHODS: Wavefront aberrations were measured by Shack-Hartmann (SH) aberrometry in eight patients with RP who had PSC cataracts, 10 patients with RP who had minimal or no PSC cataracts, and 16 age-equivalent visually normal control subjects. Higher-order wavefront aberrations for 3-mm and 6-mm pupil diameters were defined as the root mean square (RMS) of the wavefront aberration functions. RESULTS: For a 6-mm pupil diameter, the mean RMS for total higher-order wavefront aberrations was significantly larger for the patients with RP than for the control subjects, both for patients with PSC cataract (F = 17.30, p < 0.001) and for those with minimal or no PSC cataract (F = 4.80, p < 0.05). The mean RMS for third-order aberrations was not significantly different for patients with RP than for the control subjects. However, the mean RMS for fourth order aberrations was significantly larger for the patients with RP than for the control subjects, both for patients with PSC cataract (F = 8.85, p < 0.01) and those with minimal or no PSC cataract (F = 5.07, p < 0.05). There were no significant differences in higher-order aberrations between the patients with RP and the control subjects for a 3-mm pupil diameter. CONCLUSIONS: Increased higher order wavefront aberrations were present in patients with RP with and without clinically observable PSC cataracts. The measurement of wavefront aberrations in patients with RP provides an objective and quantitative method for detecting and monitoring disease-related changes in the optics of the eye. PMID- 16044076 TI - The association between eye color and corneal sensitivity measured using a Belmonte esthesiometer. AB - PURPOSE: The purpose of this study is to determine the association between corneal sensitivity measured using a pneumatic esthesiometer and eye color quantified objectively. METHODS: Twenty subjects had ocular surface sensitivity measured using a Belmonte esthesiometer. An ascending method of limits followed by the method of constant stimuli were used to estimate 1) cold detection thresholds, 2) discomfort detection thresholds (both using pneumatic stimuli at 20 degrees C, 3) mechanical detection thresholds using pneumatic stimuli at 50 degrees C (ocular surface temperature approximately 33 degrees C), and 4) percent CO2 chemical detection thresholds using 50 degrees C pneumatic stimuli at flow rates set at half of each subject's pneumatic detection threshold (therefore detected by the chemical content and not the mechanical content). Eye color was estimated 1) clinically by two observers ranking the color (light to dark) of digital images of each subject's iris, 2) photometrically by measuring iris luminance, and 3) using chromaticity obtained from a Photo Research 650 spectroradiometer with controlled illumination. Correlation and linear and nonlinear regression analyses were used to examine relationships between variables. RESULTS: There were no associations between eye color (determined clinically or objectively) for mechanical and chemical detection thresholds (best r = 0.15, all p > 0.05). There was a significant linear association between 20 degrees detection thresholds and eye color (r = 0.39), which was substantially improved with a two-line function (part level and part increasing linearly, r = 0.65). CONCLUSIONS: We were generally unable to demonstrate the relationship between eye color and sensitivity reported previously using a Cochet-Bonnet esthesiometer. However, for a subset of subjects with palest irises, there appears to be a linear association between eye color and sensitivity to cooling stimuli. PMID- 16044077 TI - Error cells for spherical powers in symmetric dioptric power space. AB - PURPOSE: : The purpose of this article is to analyze the geometry and examine the implications of the error cells of purely spherical powers in symmetric dioptric power space. METHODS: : In the context of spherocylindrical data spherical data typically implies a cylindrical component that is less than some particular amount (often 0.125 D) in magnitude. This error or uncertainty in cylinder is over and above the error in sphere itself. The two components of error are used to define the error cells in symmetric dioptric power space. RESULTS: : Error cells of spherical powers are constructed and presented as stereopairs. They are also shown in relation to error cells of powers in general. CONCLUSIONS: : An understanding of error cells can help the researcher avoid pitfalls in the analysis of spherocylindrical data. Perhaps surprisingly, the error cells of spherical powers are not invariant under spherocylindrical transposition. PMID- 16044078 TI - Nicorandil attenuates NF-kappaB activation, adhesion molecule expression, and cytokine production in patients with coronary artery bypass surgery. AB - Nicorandil (NCR), a KATP channel opener, has been reported to preserve microvascular integrity in patients with reperfused myocardial infarction. We tested the hypothesis that NCR suppresses myocardial ischemia and reperfusion injury via the attenuation of cytokine production. Forty patients who underwent coronary artery bypass graft surgery were studied. The patients were randomly divided into two groups, i.e., the patients with NCR (4-6 mg/h; N group, n = 20) or without NCR (C group, n = 20). Cardiac surgery was performed under anesthesia using fentanyl and propofol. Blood were sampled at the time of induction of anesthesia, pre-cardiopulmonary bypass, 60 min after aortic occlusion, and 60, 120, and 180 min after declamping the aorta. The activation of NF-kappaB, expression of adhesion molecules, and cytokine production were evaluated in blood samples from the control volunteers by flow cytometric analysis with or without lipopolysaccharide (LPS) stimulation in vitro. Serum IL-6 and IL-8 levels in both groups increased 60 min after declamping the aorta compared with the preoperative value (P < 0.001); the increases of these parameters in N group were lower than those in C group (P < 0.05). Serum creatine kinase with muscle and brain subunits and troponin-T levels increased 60 min after declamping the aorta in two groups (P < 0,001), but the increases of both parameters in N group were lower than those in C group (P < 0.05). NF-kappaB activation, CD11b/CD18 expression, and the production of TNF-alpha, IL-8, and IL-6 in monocytes and granulocytes were inhibited by NCR in vitro. NCR suppressed the increase of inflammatory cytokines such as IL-6 and IL-8 levels, and reduced myocardial reperfusion injury. The inhibition on NF-kappaB activation, adhesion molecule expression, and cytokine production may be one of the important mechanisms of myocardial protection of NCR. PMID- 16044079 TI - Blood neutrophil bactericidal activity against methicillin-resistant and methicillin-sensitive Staphylococcus aureus during cardiac surgery. AB - Whether methicillin-resistant Staphylococcus aureus (MRSA) constitutes per se an independent risk factor for morbidity and mortality after surgery as compared with methicillin-sensitive Staphylococcus aureus (MSSA) remains a subject of debate. The aim of this study was to assess whether innate defenses against MRSA and MSSA strains are similarly impaired after cardiac surgery. Both intracellular (isolated neutrophil functions) and extracellular (plasma) defenses of 12 patients undergoing scheduled cardiac surgery were evaluated preoperatively (day 0) and postoperatively (day 3) against two MSSA strains with a low level of catalase secretion and two MRSA strains with a high level of catalase secretion, inasmuch as SA killing by neutrophils relies on oxygen-dependent mechanisms. After surgery, an increase in plasma concentration of IL-10, an anti-inflammatory cytokine able to inhibit reactive oxygen species secretion and bactericidal activity of neutrophils, was evidenced. Despite the fact that univariate analysis suggested a specific impairment of neutrophil functions against MRSA strains, two way repeated-measures ANOVA failed to demonstrate that the effect of S. aureus phenotype was significant. On the other hand, an increase in type-II secretory phospholipase A2 activity, a circulating enzyme involved in SA lysis, was evidenced and was associated with an enhancement of extracellular defenses (bactericidal activity of plasma) against MRSA. Overall, cardiac surgery and S. aureus phenotype had a significant effect on plasma bactericidal activity. Cardiac surgery was characterized by enhanced antibacterial defenses of plasma, whereas neutrophil killing properties were reduced. The overall effect of S. aureus phenotype on neutrophil functions did not seem significant. PMID- 16044080 TI - Injury-associated hypothermia: an analysis of the 2004 National Trauma Data Bank. AB - Severe injury and shock are frequently associated with abnormalities in patient body temperature. Substantial increases in mortality have been associated with profound hypothermia, especially below 35 degrees C. The purpose of this study was to further characterize the impact of hypothermia in a large dataset of trauma patients. This study was a retrospective analysis of the 2004 version of the National Trauma Data Bank (NTDB), which contains approximately 1.1 million patients from over 400 trauma centers. Admission temperature was analyzed with respect to mortality, injury severity score (ISS), base deficit (BD), Glasgow Coma Score (GCS), and hospital outcomes. The NTDB contained 701,491 patients with temperatures recorded upon trauma center admission. Of these, 11,026 patients had admission temperatures <35 degrees C, and 802 had temperatures <32 degrees C. Comparison of core temperature versus mortality revealed that as temperature decreased, the mortality rate increased, reaching approximately 39% at 32 degrees C, and remained constant at lower temperatures. Surprisingly, 477 patients (59.5%) survived with temperatures <32 degrees C. Similarly, BD increased as hypothermia worsened until body temperature reached 31 degrees C, below which there was little further increase. Patients with admission temperatures less than 35 degrees C had significantly greater mortality (25.5% vs. 3.0%, P < 0.001) and BD (7.8 vs. 3.7, P < 0.001) when compared with patients with temperatures >or=35 degrees C. In survivors, average ventilator days and intensive care unit (ICU) days were 14.4 and 12.8, respectively, for patients with temperatures <35 degrees C as opposed to more normothermic patients who demonstrated an average of 9.5 ventilator days and 9.1 ICU days (P < 0.001). When grouped by individual ISS, BD level, and GCS motor score, mortality was significantly greater when admission temperature was below 35 degrees C (ISS mean difference = 11.4%, BD mean difference = 22.8%, and GCS motor mean difference = 9.85%). Logistic regression revealed that hypothermia remains an independent determinant of mortality after correction for confounding variables (odds ratio = 1.54, 95% confidence interval 1.40-1.71). Admission hypothermia is associated with greater mortality, increased injury severity, more profound acidosis, and prolonged ICU/ventilator courses. However, although mortality at <32 degrees C is high, patients with temperatures this low do survive. As temperatures drop below 32 degrees C, mortality rates remain constant, which may indicate a threshold below which physiologic mechanisms are unable to correct body temperature regardless of injury severity. Although shock severity is highly indicative of outcome, hypothermia independently contributes to the substantial mortality associated with severe injury. PMID- 16044081 TI - Nonspecific increase of systemic neuron-specific enolase after trauma: clinical and experimental findings. AB - The aim of this clinical and experimental study was to determine whether systemic neuron-specific enolase (NSE) is a useful early marker of traumatic brain injury (TBI) and whether NSE is affected by ischemia/reperfusion damage of abdominal organs. Our study included patients with and without TBI (verified by computerized tomography) admitted within 6 h after trauma and male Sprague-Dawley rats with ischemia and reperfusion of the abdominal organs liver, gut, or kidney. Thirty-eight study patients included 13 with isolated TBI and 18 patients with multiple trauma and TBI. Seven patients had multiple trauma but no TBI. Fifteen rats were anaesthetized and subjected to isolated ischemia of the liver, gut, or kidney (n = 5 each) for 1 h, followed by reperfusion for 3 h. In patients, NSE increased over 2-fold versus the upper normal limit (10 microg/L) within 6 h after trauma, regardless of whether TBI had occurred or not. In rats, NSE increased over 3-fold versus laboratory controls during ischemia of the liver and kidney (both P < 0.0005), but not of the gut. NSE increased over 2-fold after onset of reperfusion of the liver and kidney (both P < 0.05), but not of the gut and increased over 3-fold after 3 h of reperfusion of the liver, gut (both P < 0.005), and kidney (P < 0.0005). Our data show that systemic NSE increases to similar degrees with and without TBI. Therefore, NSE is not a useful early marker of TBI in multiple trauma. PMID- 16044082 TI - Gene expression profiles in hypoxic preconditioning using cDNA microarray analysis: altered expression of an angiogenic factor, carcinoembryonic antigen related cell adhesion molecule 1. AB - Hypoxic preconditioning has been shown to exhibit cardioprotective effects on myocardium from ischemic or reperfusion injury. The specific regulated gene involved in the hypoxia-induced cardioprotective effects is profiled in this study. Young male Wistar rats and ICR mice were exposed to sea level (as normal control) or simulated high altitude for 15 h/day for 2, 4, or 8 weeks, or for 4 weeks at high altitude after 2 weeks at sea level. The left ventricles of the animals were isolated for mRNA isolation and cDNA microarray analysis. Our data demonstrated that hypoxic preconditioning significantly ameliorated cardiac ischemic injury by minimizing the infarct size. After cluster analysis of expression profiles after different courses of hypoxic preconditioning (0, 2, 4, and 8 weeks), 386 genes showed an ascending pattern, whereas 301 genes showed a descending pattern. The ascending genes include several angiogenic factors: FGF receptor 4, vascular endothelial growth factor (vEGF), and carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM-1). The microvessel density was also significantly increased in hypoxic hearts. Using Western blotting and immunohistochemical analysis, the protein expression level and localization of CEACAM-1 were observed in hypoxic myocardium. The results also indicated that CEACAM-1 was upregulated as with other hypoxic angiogenic factors, heme oxygenase 1 (HO-1) and hypoxia inducible factor-1alpha (HIF-1alpha), in in vitro cultured cardiomyocytes (H9c2) after hypoxia treatment and in vivo hypoxic preconditioning. Furthermore, incubation with recombinant vEGF could also increase the expression level of CEACAM-1 in H9c2 cells. These results demonstrated that hypoxic preconditioning resulted in transcriptional changes, and some of these genes have been correlated with angiogenesis. The HIF 1/vEGF/CEACAM-1 pathway might be important for hypoxia-induced angiogenesis in the heart during hypoxic preconditioning. PMID- 16044083 TI - The evolution of isolated bilateral lung contusion from blunt chest trauma in rats: cellular and cytokine responses. AB - Lung contusion is the leading cause of death from blunt thoracic trauma in adults, but its mechanistic pathophysiology remains unclear. This study uses a recently developed rat model to investigate the evolution of inflammation and injury in isolated lung contusion. Bilateral lung contusion with minimal cardiac trauma was induced in 54 anesthetized rats by dropping a 0.3-kg hollow cylindrical weight onto a precordial shield (impact energy, 2.45 Joules). Arterial oxygenation, pressure-volume (P-V) mechanics, histology, and levels of erythrocytes, leukocytes, albumin, and inflammatory mediators in bronchoalveolar lavage (BAL) were assessed at 8 min, at 4, 12, 24, and 48 h, and at 7 days after injury. The role of neutrophils in the evolution of inflammatory injury was also specifically studied by depleting these cells with intravenous vinblastine before lung contusion. Arterial oxygenation was severely reduced at 8 min to 24 h postcontusion, but became almost normal by 48 h. Levels of erythrocytes, leukocytes, and albumin in BAL were increased at 0.2 would include all patients with an aberrant genotype but would result in a reduction of 52% of genotyping reactions. Slow metabolism of amitriptyline is correlated with a log(MR) > 0.4. Genotyping only those subjects with a log(MR) > 0.4 would result in 88% fewer genotyping reactions. For risperidone, genotyping individuals with a log(MR) > 0.4 would include all CYP2D6 poor metabolizers while reducing the number of genotyping reactions by 93%. According to these data, correlations exist between the log(MR) of venlafaxine, amitriptyline, and risperidone and the genotype of the CYP enzymes involved in their metabolism. From the ranges of log(MR) defined here, a high percentage of aberrant metabolizers can be detected even when patients are not routinely genotyped. Thus, the metabolic ratio may serve as an indication of when genotyping should be considered. PMID- 16044106 TI - Resistance to aspirin in patients after coronary artery bypass grafting is transient: impact on the monitoring of aspirin antiplatelet therapy. AB - The aim of the present study was to assess the responsiveness of blood platelets to aspirin in patients following coronary artery bypass grafting (CABG) surgery. Aspirin was administered following CABG in 24 operated patients (aged 63.2 +/- 6.3 years). Platelet function was monitored on the 10th postoperative day (A) and 1 month after CABG (B) with the use of whole-blood aggregometry (WBEA) and PFA 100 closure time (PFA-100 CTCEPI). Normal platelet response to aspirin was defined by 3 criteria: the complete inhibition of WBEA induced by arachidonic acid (0.5 mmol/L), partial inhibition of collagen (1 microg/mL)-induced aggregation (WBEA < 14 Omega), and prolongation of PFA-100 CTCEPI (>150 seconds) ("good responders"). Depending on whether 0, 1, 2, or 3 of these 3 criteria were fulfilled, patients were classified as "nonresponders," "weak responders," "incomplete responders," or "good responders," respectively. On the 10th postoperative day, there were 3 good responders, 6 incomplete responders, 11 weak responders, and 4 nonresponders among 24 patients. In contrast, 1 month after CABG within the same group of 24 patients there were 18 good responders, 5 incomplete responders, and 1 weak responder. Using a new methodology to assess impaired platelet responsiveness to aspirin ex vivo, we describe here the transient nature of "aspirin resistance" following CABG. These results indicate the necessity for the prolonged monitoring of the antiplatelet effectiveness of aspirin in the postoperative period after CABG. PMID- 16044107 TI - Inter- and intraindividual variabilities in pharmacokinetics of fentanyl after repeated 72-hour transdermal applications in cancer pain patients. AB - Perception of pain by the patient is frequently one of the early signs preceding a diagnosis of cancer and, later, a sinister sign of disease progression. Among opioid drugs, transdermal fentanyl has been evaluated in the treatment of moderate to severe cancer pain. The objective of this study was to investigate the intra- and interindividual variabilities in pharmacokinetics after fentanyl drug delivery by the transdermal fentanyl patch delivery system in patients with cancer pain. As a first step, a liquid chromatography-mass spectrometry method was developed for the determination of the analgesic fentanyl in human plasma. This method was validated over the concentration range 0.15-100 ng/mL. The study group consisted of 29 inpatients (18 men and 11 women; age range 29-80 years). The initial transdermal fentanyl delivery rate was chosen depending on the patient's analgesic requirements. For 20 patients, the initial TTS fentanyl delivery rate was 25 or 50 microg/h. For 6 patients, the initial delivery rate was 75-150 microg/h. Two patients received up to 300 microg/h fentanyl delivery rate, and 3 patients received up to 350 microg/h fentanyl delivery rate. Fifteen of the 29 patients received rescue doses of subcutaneous or oral morphine, and 26 patients received paracetamol with codeine (30 mg per os). Blood samples were collected at the following intervals: 2-5, 22-26, or 45-47 hours following fentanyl patch application. The severity of pain experienced by the patient was assessed thrice daily using a visual analogue scale. The study period was 46 days. Large patient-to-patient variations in pharmacokinetic parameters occurred, although intraindividual variability was limited. A mean bioavailability of 78% was estimated; the total clearance averaged 41 L/h. From 25 to 100 mug/h fentanyl delivery rate, the pharmacokinetics was linear. At the 2 highest doses, an increase of total clearance was observed (>60 L/h). For the whole group, transdermal fentanyl treatment provided good to excellent pain relief in the majority of patients. PMID- 16044108 TI - The safety of amoxicillin/clavulanic acid and cefuroxime during lactation. AB - Breast-feeding is considered the gold standard for infant nutrition. In spite of statements about the safe use of drugs in lactation by the American Academy of Pediatrics, medical professionals remain confused regarding the management of drug therapy in nursing mothers, and this can lead to suboptimal prescribing and poor compliance. The aim of our study was to evaluate the safety of 2 of the newer antibiotics, amoxicillin/clavulanic acid and cefuroxime, during lactation. Breast-feeding women who called a drug consultation center to obtain information about the potential risks of amoxicillin/clavulanic acid (67 women) and cefuroxime (38 women) were prospectively recruited. As a control group, women who were treated with antibiotics known to be safe during lactation were recruited: amoxicillin (n = 40) for the amoxicillin/clavulanic acid group and cephalexin (n = 11) for the cefuroxime group. Women in the control group were matched for indication for antibiotic therapy, duration of treatment, and maternal age. Participants were interviewed after treatment termination regarding adverse reactions during therapy. In the amoxicillin/clavulanic acid group, 15 infants (22.3%) had adverse effects, and the rate increased with dosage (P = 0.0139). This was significantly higher than the amoxicillin group, where 3 infants (7.5%) had adverse effects (P = 0.046, relative risk (RR) = 2.99, 95% confidence interval (CI) 0.92-9.68). However, there were no significant differences between rates of specific events. The rate of adverse effects in the cefuroxime group (2.6%) was not significantly different from that in controls (9%) (P = 0.58, OR = 0.92, 95% CI 0.94-1.06). All adverse effects were minor, self-limiting, and did not necessitate interruption of breast-feeding. Our data suggest that amoxicillin/clavulanic acid and cefuroxime may be safe during lactation. Larger studies are needed to confirm these findings. PMID- 16044109 TI - Development and validation of a solid-phase extraction-liquid chromatographic method for determination of amoxicillin in plasma. AB - A bioanalytic method for the determination of amoxicillin in plasma by hydrophilic interaction solid-phase extraction and liquid chromatography has been developed and validated. Plasma was precipitated with acetonitrile before samples were loaded onto a zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) solid-phase extraction column. Amoxicillin was analyzed by liquid chromatography on an Aquasil (150 x 4.6 mm) LC column with mobile-phase acetonitrile: phosphate buffer (pH 2.5; 0.1 mol/L) (7:93, v/v) and UV detection at 230 nm. A regression model using 1/concentration weighting was found the most appropriate for quantification. The intraassay precision for plasma was 3.3% at 15.0 microg/mL and 10.9% at 0.200 microg/mL. The interassay precision for plasma was 1.8% at 15.0 microg/mL and 7.5% at 0.200 microg/mL. The total-assay precision for plasma over 4 days using a total of 20 replicates was 13.2%, 5.5%, and 3.8% at 0.200 microg/mL, 3.00 microg/mL, and 15.0 microg/mL, respectively. The lower limit of quantification and the limit of detection were 0.050 microg/mL and 0.025 microg/mL, respectively, for 100 microL plasma. Long-term storage stability studies of amoxicillin in plasma indicate that a temperature of -80 degrees C is necessary to prevent degradation of amoxicillin. PMID- 16044110 TI - Screening procedure for detection of diuretics and uricosurics and/or their metabolites in human urine using gas chromatography-mass spectrometry after extractive methylation. AB - A gas chromatography-mass spectrometry (GC-MS)-based screening procedure was developed for the detection of diuretics, uricosurics, and/or their metabolites in human urine after extractive methylation. Phase-transfer catalyst remaining in the organic phase was removed by solid-phase extraction on a diol phase. The compounds were separated by GC and identified by MS in the full-scan mode. The possible presence of the following drugs and/or their metabolites could be indicated using mass chromatography with the given ions: m/z 267, 352, 353, 355, 386, and 392 for thiazide diuretics bemetizide, bendroflumethiazide, butizide, chlorothiazide, cyclopenthiazide, cyclothiazide, hydrochlorothiazide, metolazone, polythiazide, and for canrenoic acid and spironolactone; m/z 77, 81, 181, 261, 270, 295, 406, and 438 for loop diuretics bumetanide, ethacrynic acid, furosemide, piretanide, torasemide, as well as the uricosurics benzbromarone, probenecid, and sulfinpyrazone; m/z 84, 85, 111, 112, 135, 161, 249, 253, 289, and 363 for the other diuretics acetazolamide, carzenide, chlorthalidone, clopamide, diclofenamide, etozoline, indapamide, mefruside, tienilic acid, and xipamide. The identity of positive signals in such mass chromatograms was confirmed by comparison of the peaks underlying full mass spectra with reference spectra. This method allowed the detection of the abovementioned drugs and/or their metabolites in human urine samples, except torasemide. The limits of detection ranged from 0.001 to 5 mg/L in the full-scan mode. Recoveries of selected diuretics and uricosurics, representing the different chemical classes, ranged from 46% to 99% with coefficients of variation of less than 21%. After ingestion of the lowest therapeutic doses, furosemide was detectable in urine samples for 67 hours, hydrochlorothiazide for 48 hours, and spironolactone for 52 hours (via its target analyte canrenone). The procedure described here is part of a systematic toxicological analysis procedure for acidic drugs and poisons. PMID- 16044111 TI - Determination of nevirapine and efavirenz in plasma using GC/MS in selected ion monitoring mode. AB - An accurate, selective, and sensitive method for the determination of the nonnucleoside reverse transcriptase inhibitors (NNRTIs) nevirapine (nvp) and efavirenz (efv) in human plasma using gas chromatography-mass spectroscopy in selected ion monitoring mode (GC/MS-SIM) was developed. Solid-phase extraction (SPE) gave extraction yields near 100% for both nvp and efv, and calibration curves were linear over the therapeutic concentration ranges. Variation of intraday and interday precision was below 5%. Intraday and interday inaccuracies varied between 0.6% and 10.4%. The lower limits of detection using a 200-microL plasma sample were 27 ng/mL for nvp and 26 ng/mL for efv, and the lower limits of quantification were 54 ng/mL and 72 ng/mL, respectively. The method was applied to the determination of nvp and efv in plasma specimens of 73 patients in HIV stage III or IV and on antiretroviral treatment in Kigali, Rwanda. PMID- 16044112 TI - Buprenorphine and norbuprenorphine concentrations in human breast milk samples determined by liquid chromatography-tandem mass spectrometry. AB - Buprenorphine (BUP) is considered to be safe during pregnancy. However, the extent of BUP transfer into breast milk has not been investigated thoroughly. Because the drug concentration in the milk is 1 of the determinants in the assessment of the exposure risk, a rapid and sensitive LC-MS/MS method has been developed and evaluated to measure BUP and norbuprenorphine (norBUP) concentrations in milk. A solid-phase and 2 liquid-liquid extraction procedures have been compared. The lower limits of detection and quantification were 0.05 ng/mL and 0.18 ng/mL for BUP and 0.05 ng/mL and 0.20 ng/mL for norBUP, respectively, using a sample volume of 0.5 mL milk. BUP and norBUP concentrations determined from 10 random breast milk samples collected over 4 successive days from a lactating woman during buprenorphine maintenance therapy ranged from 1.0 to 14.7 and 0.6 to 6.3 ng/mL, respectively. Drug exposure of the infant may be considered to be low. Further investigations may seek to extend these preliminary findings to evaluate an infant's level of BUP exposure through breast milk. PMID- 16044113 TI - MDR1 genetic polymorphisms and comparison of MDR1 haplotype profiles in Korean and Vietnamese populations. AB - Two representative genetic variants of the MDR1 gene, 3435C>T and 2677G>T/A, show wide interethnic differences in its genetic polymorphism. In this study, the authors evaluated the genetic polymorphisms of MDR1 and directly compared MDR1 haplotype profiles of the Korean and Vietnamese populations. The 3435C>T and 2677G>T/A variations were analyzed in 632 Koreans and 142 Vietnamese using pyrosequencing. The allelic frequencies of 3435C>T did not significantly differ between the Korean (39.3%) and Vietnamese (36.6%) groups. However, the frequencies of mutant alleles at 2677 locus (T or A allele) showed a significant difference between Koreans (56.2%) and Vietnamese (41.9%), as the frequency of 2677A allele in the Korean subjects (17.1%) was much higher than that of the Vietnamese subjects (6.3%). Linkage analysis revealed that 2677A allele is closely linked to 3435C allele. The frequency of 2677A-3435C haplotype in Koreans was 15.4%, which was significantly higher than that found in Vietnamese subjects (6.3%). In conclusion, the frequencies of MDR1 variants and haplotype profiles showed significant differences between the Korean and Vietnamese populations, especially with respect to the 2677G>T/A variants. Because the 2677A allele was recently found to be functional in vivo and was detected at a high frequency in Koreans, the genotyping of this variant is necessary for pharmacogenetic studies of MDR1 in this population. In addition, by virtue of strong linkage disequilibrium, 2677A-3435C haplotype may help improve the predictability of MDR1 genetic polymorphism for MDR1 functional changes. PMID- 16044114 TI - Pharmacokinetic changes of irinotecan by intestinal alkalinization in an advanced colorectal cancer patient. AB - The prevention of irinotecan (CPT-11)-induced diarrhea, a well-known adverse reaction to the drug, by treatment with intestinal alkalinization has been carried out in patients with colorectal cancer in Japan. Under acidic conditions, CPT-11 and its active metabolite, SN-38, exists preferably as the lactone form, whereas both exist as the carboxylate form under basic conditions. It has been suggested that the lactone forms of both CPT-11 and SN-38 are diffused passively across the intestinal mucosal membranes, whereas the carboxylate forms are actively transported. The intestinal uptake rate of both forms appears to be pH sensitive under physiological conditions, but it remains unclear whether intestinal alkalinization treatment affects the pharmacokinetics of CPT-11 and SN 38. This study was designed to evaluate the pharmacokinetics of CPT-11 and SN-38 in a colorectal cancer patient with or without alkalinization treatment. We found that intestinal alkalinization significantly decreased the plasma levels of CPT 11 and SN-38. In particular, the AUC of SN-38 was markedly decreased to 56 from 107 ng.h/mL. Intestinal alkalinization was effective in preventing CPT-11-induced diarrhea, but this treatment changed the pharmacokinetics of CPT-11 and SN-38 in the body. PMID- 16044115 TI - Increased clozapine plasma concentrations and side effects induced by smoking cessation in 2 CYP1A2 genotyped patients. AB - Clozapine, an atypical antipsychotic, depends mainly on cytochrome P4501A2 (CYP1A2) for its metabolic clearance. CYP1A2 is inducible by smoking, and lower plasma concentrations of clozapine are measured in smokers than in nonsmokers. Case reports have been published on the effects of discontinuing smoking in patients receiving clozapine, which might lead to elevated plasma concentrations and severe side effects. We present 2 cases on the consequences of smoking cessation in patients receiving this drug. In the first patient, smoking cessation resulted, within 2 weeks, in severe sedation and fatigue, with an approximately 3-fold increase of plasma clozapine concentrations. In the second patient, a very high plasma concentration of clozapine (3004 ng/mL) was measured 6 days following a 16-day stay in a general hospital, during which smoking was prohibited. In the latter patient, the replacement of omeprazole, a strong CYP1A2 inducer, by pantoprazole, a weaker CYP1A2 inducer, could have contributed, in addition to smoking cessation, to the observed strong increase of plasma clozapine concentrations. Genotyping of the 2 patients revealed that they were carriers of the AA genotype for the -164C>A polymorphism (CYP1A2*1F) in intron 1 of CYP1A2 gene, which has previously been shown to confer a high inducibility of CYP1A2 by smoking. Thus, at the initiation of clozapine treatment, smoking patients should be informed that, if they decide to stop smoking, they are encouraged to do so but must inform their prescriber beforehand. Also, because of the increased use of no-smoking policies in many hospitals, studies examining the consequences of such policies on the pharmacokinetics/pharmacodynamics of drugs metabolized by CYP1A2, taking into account different CYP1A2 genotypes, are needed. PMID- 16044117 TI - [Treatment of patients with general surgical diseases after open-heart operations with cardiopulmonary bypass]. AB - Experience in surgical treatment of 69 patients suffering from coronary heart disease and valve pathology in combination with various general surgical diseases was summarized. Two methods of surgical correction were studied: combined one and staged one. Up-to-date technologies such as endoscopic surgery, tension free hernioplasty, and artificial materials were used during the procedures. Combined procedures reduce the risk of general surgery complications early after surgery. Correction of cardiovascular pathology has a priority in curative algorithm. PMID- 16044118 TI - [Sclerotherapy of high venovenous blood reflux in patients with early varicose disease of the lower extremities]. AB - Results of diagnosis and treatment of 270 patients with varicose disease were analyzed. Based on duplex scanning two groups of patients were divided: group 1 consisted of 164 patients with low-intensity reflux, group 2 consisted of 106 patients with high-intensity reflux. Patients of group 1 were treated with ultrasound-assisted puncture sclerotherapy. In group 2 sclerosurgical procedures were used: sclerosant was injected into great saphenous vein trough catheter after crossectomy. For obliteration of varicose subcutaneous veins and perforants of shank the Irish technique was used. In group 1 recurrence of the disease was revealed in 9.1% patients, in group 2 -- in 9.4% patients. PMID- 16044119 TI - [Disputable points and negative tendencies in diagnosis and surgical treatment of accidentally revealed adrenal tumors]. AB - Two main problems are discussed in detail: differential diagnosis of adrenal tumors before surgery, and policy of treatment of these patients. Indications for fine needle biopsy are determined, based on personal and world experience reasoned indications to surgical treatment and follow-up are formulated. Some negative tendencies in adrenal surgery are also discussed. PMID- 16044120 TI - [Surgical and combined treatment of multiple and bilobar metastatic affection of the liver]. AB - Short- and long-term results of surgical treatment in combination with local exposures (cryo- and thermo-destruction) and adjuvant chemotherapy in patients treated for multiple and bilobar metastatic affection of the liver were studied. Postoperative lethality was 2,5% (n=2) due to liver insufficiency. Specific for liver resection complications (bile outflow, moderate hepatic failure) do not exceed 20,5%. Five-year survival of patients with solitary bilobar metastatic affection of the liver (not more than two foci) was 39,1+/-14,3%. This demonstrates that solitary bilobar hepatic metastases is not a negative prognostic factor for surgical treatment. Long-term results of treatment of patients with multiple metastases of colorectal cancer to the liver are worse, but 1-, 2-, and 3-year survival demonstrates validity of surgical policy in treatment of these patients. Postoperative chemotherapy significantly improves long-term survival of patients with multiple or/and bilobar metastases of colorectal cancer to the liver. PMID- 16044121 TI - [Surgical approach for destructive cholecystitis in elderly and old patients]. AB - The individual surgical policy in the treatment of patients over 60 years of age with destructive cholecystitis was developed. Urgent radical surgical procedures using total intravenous anesthesia with endotracheal intubation and ALV were performed in patients with a low surgical and anesthetic risk and without concomitant acute pancreatitis and obstructive jaundice. Cholecystostomy and delayed cholecystectomy were performed in patients with these concomitant pathologies. Palliative operations were performed in patients with high surgical and anesthetic risk. Patients with disseminated peritonitis underwent cholecystectomy through laparotomy using total intravenous anesthesia with epidural blockade. Choice of method of cholecystectomy and anesthetic management depended on nature of concomitant diseases and complications. Proposed individual surgical policy permitted to decrease postoperative lethality to 0.8%. PMID- 16044122 TI - [Cost-effect evaluation of different variants of cholecystectomies]. AB - Short-term results of treatment and cost-effect rates were analyzed in three groups of patients undergone traditional cholecystectomy, laparoscopic cholecystectomy and cholecystectomy through minilaparotomy. It was demonstrated that frequency of complications was less after operations through laparoscopy and minilaparotomy. Traditional approach had the most costs due to more number of postoperative complications, hospital stay after surgery and more number of analgesics. Cholecystectomy through minilaparotomy is most cost-effective due to high clinical effectiveness, short hospital stay and rare complications. This procedure does not require expensive devices and can be performed with regional anesthesia only. It is concluded that minimally invasive methods of cholecystectomy must be used more widely. PMID- 16044123 TI - [Early cancer of the stomach: pattern of lymphatic metastasizing and its influence on surgical approach]. AB - Results of surgical treatment of 156 patients were analyzed. There were no metastases to the lymph nodes in patients with invasion of mucous membrane only. In patients with invasion of the tumor into a submucous layer regional metastases were revealed in 18 (19%) cases. Number of affected lymph nodes varied from 1 to 7 (2.7, on the average). Metastases to 1 lymph node were revealed in 9 (50%) patients, to 2-3 nodes -- in 5 patients, to 5-7 nodes -- in 4 patients. In 14.78% patients metastases were only to perigastric lymph nodes, in 4 cases -- to nodes of the second level (N2). Invasion of the tumor into submucous layer, location and size of the tumor were the main factor of risk of lymphatic cancer spread. Frequency of lymphatic cancer spread does not exceed 10% in any variant. In patients with proximal tumor, infiltrative growth and size more than 4 cm, tumor lymphatic metastases are diagnosed in each third person. It is concluded that gastrectomy or distal subtotal resection of the stomach with modified D2 lymphatic dissection is the method of choice in treatment of patients with invasion of stomach cancer into submucous layer. PMID- 16044124 TI - [Surgical treatment of diseases of operated stomach]. AB - Experience with treatment of 262 patients with late complications after operations for duodenal ulcer was analyzed. Eighty-four (32.1%) patients underwent earlier suturing of perforated duodenal ulcer, 97 (37.0%) -- resection of the stomach and 81 (30.9%) -- various types of vagotomy. In recurrent ulcers after sutured perforation organ-saving operations (62, or 73.8%) have important advantages over resection of the stomach. Combined postgastrectomy syndromes (18, or 18.6%) were most difficult for surgical treatment. Resection of the stomach (70, or 86.4%) patients is the best reconstructive operation for postvagotomy syndromes. PMID- 16044125 TI - [Primary infarction of the greater omentum]. AB - Results of treatment of 10 patients with primary infarction of the greater omentum were analyzed. Young and middle-aged persons prevailed. Majority of the patients were hospitalized from 27 to 72 hours after disease manifestation. The cause of disease was not revealed in all the patients. Pains in right half of the abdomen were the main symptoms of primary infarction of the greater omentum. There were no specific clinical or laboratory signs. Seven patients were operated for suspected acute appendicitis, and 3 patients - for suspected acute cholecystitis. Pathology of the greater omentum was combined with hemorrhagic effusion in 8 of 10 patients. Resection of affected part of the greater omentum was performed in all the patients, 6 patients underwent simultaneous appendectomies. There were no postoperative complications. All the patients recovered. Mistakes leading to incorrect diagnosis are analyzed. PMID- 16044126 TI - [Modification of rehabilitation stage after inter-sphincter resection of the rectum]. AB - Endoscopic ultrasonic examination for diagnosis of malignant tumors of a lower ampullar part of the rectum determined indications to inter-sphincter resection consisting in total mesorectumectomy, resection of the rectum and internal sphincter. The method of creation of a globe-shape reservoir with artificial muscular sphincter was developed. Fourteen patients underwent surgery, the follow up was up to 3,5 years. It is demonstrated that a globe-shape extensible reservoir and autonomic bloching function of muscular sphincter are the main conditions of repair of defecation functions in long-term period after inter sphincter resection. PMID- 16044127 TI - [Methods of wounds closure in the treatment of open and gunshot fractures of the lower extremities]. AB - The developed methods of wounds closure were successfully used in 25 patients aged from 15 to 55 years with open and gunshot fractures of the lower extremities. The former method consists in gradual putting of wound borders on Kirschner's wires and gradual closure of wound defect. The latter method is based on using original removable plate sutures which permit to regulate degree of adaptation of wound borders. Indications and contraindications for methods were determined. PMID- 16044128 TI - [Surgical approach in bilateral and combined echinococcosis of the lung]. AB - One hundred and two patients aged from 4 to 65 years with echinococcosis were treated. Solitary cysts in both lungs were diagnosed in 69 patients, multiple cysts -- in 33 patients. In 10 cases small and middle cysts were in the upper parts of both lungs. These patients underwent one-stage operation consisting in lateral thoracotomy, removing of the cyst from one lung, incision of retrosternal mediastinal pleura, removing of the cyst from the second lung, drainage of pleural cavity. Fifteen patients with combined echinococcosis of the upper lobe of the right lung and liver were operated with another one-stage surgical method through thoracotomy in the third and sixth or seventh intercostal space. These methods are characterized by low traumaticity, decreased number of postoperative complications and hospital stay. PMID- 16044129 TI - [Ultrasonic diagnosis of choledocholithiasis]. AB - Potential of ultrasonic examination for diagnosis of choledocholithiasis was analyzed in 47 patients. Ultrasonic scanners Acuson 128XP and SONOLINE Elegra with 2-9 MHz sensors were used. Choledocholithisis was diagnosed during transabdominal echography in 38 (80.9%) patients. Method of ultrasound-assisted "vibratory" palpation for diagnosis of mobile concrements of the common bile duct is presented. Causes of 2 (4.3%) false-positive and 7 (14.9%) false-negative results of examination are analyzed. PMID- 16044130 TI - [Computed tomography diagnosis of hepatic space-occupying lesion of different nature]. AB - Specific CT-symptoms are reliable criteria for differential diagnosis of benign lesions of the liver. The method permits to diagnose exactly cystic lesions of the liver in 95.3% cases, and tumor lesions -- in 83%. Diagnostic value of CT is higher in solitary non-parasitic cysts, polycystic disease and echinococcosis than in hemangioma, alveococcosis and malignant tumors of the liver (p<0,05). Contrast CT increases differential and diagnostic value of the method. PMID- 16044131 TI - Use of DLI to achieve complete donor chimerism in a patient receiving systemic immunosuppression for refractory AIHA post-NST. PMID- 16044132 TI - Central pontine myelinolysis following bone marrow transplantation complicated by severe hepatic veno-occlusive disease. PMID- 16044133 TI - Risks and outcomes of invasive fungal infections in pediatric patients undergoing allogeneic hematopoietic cell transplantation. AB - Invasive fungal infections (IFI) are the leading cause of infectious mortality in adult patients undergoing hematopoietic cell transplantation (HCT) after myeloablative conditioning, but the extent of this problem in the pediatric population is unclear. We retrospectively examined risk factors for IFI among 120 consecutive pediatric patients undergoing allogeneic HCT at a single center. The incidence of proven or probable IFI in pediatric patients during the first year after allogeneic HCT was 13%, comparable to the rate reported in adult patients; however, unlike IFI in adult patients, the majority of IFI in children occurred within the first month after transplantation. The primary risk factors for IFI were duration of neutropenia, age greater than 10 years, transplant for severe aplastic anemia or Fanconi anemia, and high-dose corticosteroid administration for 10 days or longer. IFI were more likely to be successfully treated (42%, 5/12 patients) in pediatric HCT recipients when compared to previous reports of adult recipients. Nonrelapse mortality was estimated at 17% (20/120 patients) after allogeneic HCT, of which 35% (seven patients) were directly attributed to IFI. Thus, IFI is a significant cause of nonrelapse mortality in children undergoing allogeneic HCT and more effective strategies are needed to prevent and treat IFI. PMID- 16044134 TI - Long-term renal function after hemopoietic stem cell transplantation in children. AB - Glomerular function of all long-term survivors who underwent hemopoietic stem cell transplantation (HSCT) from 1991 to 1998 (study I, n=121) was studied retrospectively. In addition, we prospectively analyzed glomerular and tubular function of all long-term surviving children who received an HSCT between 1998 and 2000 (study II, n=41). We found a lower prevalence of children with chronic renal failure (CRF) post-HSCT in our more recent cohort (study II: 10%) as compared to the older cohort (study I: 24%) 5.0 (0.7 s.d.) and 7.6 (2.4 s.d.) year's post-HSCT, respectively. Furthermore, it seems that renal function may stabilize after 1-year post-HSCT. None of the patients required dialysis or antihypertensive medication at long-term follow-up. The sole predictor of CRF in our study was high serum creatinine pre-HSCT (P=0.007), while acute renal failure within 3 months after HSCT (P=0.08) only showed a trend towards predicting CRF. We could not confirm a relation of conditioning with irradiation with CRF post HSCT, as was shown in several other pediatric and adult studies. Proximal and distal tubular dysfunction only occurred in a minority of long-time survivors of HSCT (3-12 and 9-13%, respectively) and had no clinical consequences. PMID- 16044135 TI - Do minitransplants have minicosts? A cost comparison between myeloablative and nonmyeloablative allogeneic stem cell transplant in patients with acute myeloid leukemia. AB - Allogeneic hematopoietic stem cell transplantation (SCT) is a widely used, cost intensive procedure. Although pretransplant nonmyeloablative (NMA) or reduced intensity conditioning regimens appear very promising, prospective studies comparing this approach with the conventional myeloablative (MA) approach in specific hematologic diseases are necessary, especially in patients in whom the conventional approach is not contraindicated. Cost may be an important factor in the decision-making process. We compared the costs of MA and NMA transplants in patients with acute myeloid leukemia (AML). We estimated 1-year resource utilization in 12 consecutive MA patients (median age: 39 years) and in 11 consecutive NMA patients (median age: 58 years) who underwent HLA-identical sibling SCT for AML. Resources care expenses were valued using the average daily rate for personnel costs, supplies, and room costs. Other data were directly collected from the patients' charts. Despite a trend for lower costs in NMA patients during the first 6 months, costs during the 6-12-month period were significantly higher after NMA due to late complications and readmissions (P=0.03). Finally, mean 1-year costs were not different in MA and NMA patients (P=0.75). Prospective studies comparing conventional and NMA approaches in homogeneous populations should include economic items. PMID- 16044136 TI - IL-4-producing CD8(+) T cells may be an immunological hallmark of chronic GVHD. AB - Chronic graft-versus-host disease (cGVHD) occurs in approximately 60-80% of those who survive over 100 days after allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, the pathophysiology of cGVHD is poorly understood. To gain more insight into the immunological mechanism of cGVHD, we examine cytokine production of peripheral blood T cells from 19 patients in the chronic phase of allo-HSCT. The percentage of IFN-gamma-producing CD8(+) T cells among CD8(+) T cells was significantly higher in patients with or without cGVHD than in normal control subjects (P<0.001). On the other hand, the percentage of IL-4-producing CD8(+) T cells among CD8(+) T cells was significantly higher in patients with cGVHD (mean 3.3%; range 1.3-8.2%) than in patients without cGVHD (mean 1.2%; range 0.8-1.7%) and normal control subjects (mean 1.1%; range 0.1 1.6%) (both P<0.001). By contrast, the percentage of IL-4-producing CD4(+) T cells was not different among patients with and without cGVHD and normal controls. These findings suggest that IL-4-producing CD8(+) T cells may be an immunological marker of cGVHD. PMID- 16044137 TI - Serum free light-chain responses after high-dose intravenous melphalan and autologous stem cell transplantation for AL (primary) amyloidosis. AB - Serum free light-chain (FLC) concentrations were measured by a sensitive nephelometric immunoassay in 66 patients with AL amyloidosis before and after treatment with high-dose melphalan and autologous stem cell transplantation (HDM/SCT). At 1 year after HDM/SCT, 27 patients (41%) achieved a complete hematologic response (CR), that is, disappearance of the monoclonal gammopathy previously evident by immunofixation electrophoresis (IFE) in serum and urine and of plasma cell clonality in the bone marrow. Abnormally elevated FLC levels became normal in 27 patients (41%), and decreased by >90% in 37 (56%). Average improvements in FLC were 94% for patients who achieved a CR and 72% for those who did not (P=0.0001). However, a reduction in FLC of >90% was associated with a similar high likelihood of clinical improvement and prolonged survival, whether or not patients achieved a CR. While CR, as defined by standard criteria, is a more stringent indicator of hematologic response than are decreases in abnormally elevated FLC levels per se, these measures of hematologic response are complementary, and decreases in FLC are more readily detected early after treatment than are the changes in IFE and marrow studies required to determine CR. PMID- 16044138 TI - Increased frequency of the angiotensin-converting enzyme gene D-allele is associated with noninfectious pulmonary dysfunction following allogeneic stem cell transplant. AB - Noninfectious pulmonary dysfunction (NIPD) is a common and often fatal complication associated with allogeneic hematopoietic stem cell transplantation (HSCT). An insertion/deletion polymorphism in the angiotensin-converting enzyme (ACE) gene has been extensively studied in relation to cardiovascular and renal disease, and lung fibrosis. In pulmonary fibrosis, D-allele frequency is significantly higher than in the control population. We hypothesized that a similar mechanism exists between post-HSCT NIPD and pulmonary fibrosis in the absence of HSCT. We retrospectively analyzed the incidence of NIPD and the ACE genotype polymorphism in 118 Japanese patients who underwent HSCT from HLA identical sibling donors. NIPD occurred in 17 cases. Deletion/deletion genotype carriers were more common in the NIPD group than in the other 101 patients (41.2 vs 11.9%; hazard ratio, 5.19; 95% confidence interval, 1.67-16.21). There were no significant relationships between the clinical characteristics of patients and the development of NIPD. These findings suggest that the ACE genotype is associated with the development of NIPD following HSCT. This study is the first to report the relationship between genetic background and NIPD. PMID- 16044139 TI - Pilot study of tandem high-dose chemotherapy and autologous stem cell transplantation with a novel combination of regimens in patients with poor risk lymphoma. AB - In an effort to improve the outcome of poor-risk lymphoma patients, we evaluated a novel regimen of tandem high-dose chemotherapy (THDC) with autologous stem cell transplantation. A total of 41 patients (median age 40 years, range 15-68 years) with poor-risk non-Hodgkin's lymphoma and Hodgkin's disease were enrolled. THDC consisted of melphalan (180 mg/m2) and escalating dose mitoxantrone (30-50 mg/m2) (MMt) for the first conditioning regimen, and thiotepa (500 mg/m2), carboplatin (800 mg/m2), and escalating dose etoposide phosphate (400-850 mg/m2), (ETCb) as the second regimen. In all, 31 patients (76%) completed both transplants, with a median time between transplants of 55 days (range 26-120). The maximum tolerated dose was determined as 40 mg/m2 for mitoxantrone and 550 mg/m2 for etoposide phosphate. The overall toxic death rate was 12%. Following high-dose chemotherapy, 10 of 24 evaluable patients (42%) were in CR. The two-year overall survival and event-free survival is 67% (95% CI, 52-81%) and 45%, (95% CI, 29 61%) for the 41 patients enrolled; and 69% (95% CI, 525-586%) and 48% (95% CI, 30 67%) for the 31 patients completing both transplants. This THDC regimen is feasible but with notable toxicity in heavily pretreated patients; its role in the current treatment of high-risk lymphoma remains to be determined. PMID- 16044140 TI - What would Karl Landsteiner do? The ABO blood group and stem cell transplantation. AB - ABO blood group antigens, of great importance in transplantation and transfusion, are present on virtually all cells, as well as in soluble form in plasma and body fluids. Naturally occurring plasma IgM and IgG antibodies against these antigens are ubiquitous. Nonetheless, the ABO blood group system is widely ignored by many transfusion services, except for purposes of red cell transfusion. We implemented a policy of transfusing only ABO identical platelets and red cells in patients undergoing stem cell transplantation or treatment for hematologic malignancies. Major bleeding episodes have occurred in about 5% of patients undergoing induction therapy for acute leukemia as compared with 15-20% in the literature. Overall survival times appear to be superior to that in historical cohorts. In 2002-2004, treatment-related mortality at 100 days in our Blood and Marrow Transplant Unit was 0.7% for autologous transplants (n=148), 13% for sibling allogeneic transplants (n=110), and 24% (n=62) for matched unrelated allogeneic transplants, suggesting that our approach is safe. We speculate that more rigorous efforts on the part of transfusion services to provide ABO identical blood components, and to remove incompatible supernatant plasma, when necessary, might yield reduced morbidity and mortality in patients undergoing stem cell transplantation. PMID- 16044141 TI - Variation in dimethyl sulfoxide use in stem cell transplantation: a survey of EBMT centres. AB - The cryoprotectant dimethyl sulfoxide (DMSO) is known to have toxic side effects, yet guidelines for its use in stem cell transplantation do not exist. To assess current practice in the use of DMSO and the incidence of DMSO-related complications, a single page questionnaire was mailed to 444 EBMT centres involved in autologous transplantation. The responses from 97 centres showed a wide variation in practice between transplant units regarding the concentration of DMSO used, daily DMSO dose restriction and the use of cell washing. The overall incidence of DMSO toxicity was approximately one in 70 transplants and most cases were cardiovascular and respiratory in nature. There was a trend to reduced complication rates in centres using lower concentrations of DMSO or washing cells prior to return. A large-scale prospective study of the strategies for reduction in exposure to DMSO and reduction in toxic effects is required before guidelines in the use of DMSO in stem cell cryopreservation can be promulgated. PMID- 16044142 TI - Hourly monitoring of circulating CD34+ cells to optimize timing of autologous apheresis in pediatric patients. AB - In order to increase the CD34+ cell yield in children undergoing autologous stem cell transplantation, the optimum time of apheresis after G-CSF administration has still to be found. We prospectively studied the mobilization of CD34+ cells and white blood cells in the peripheral blood (PB) of 20 pediatric patients before leukapheresis. The monitoring schedule covered 12 h, with blood samples taken before and at 2, 4, 5, 6, 7, 8, 10 and 12 h after G-CSF administration when 10 CD34+ cells/mul were reached. CD34+ cells were measured by flow cytometric analysis both in the single- and dual-platform setting. Two different patterns of mobilization (POM) emerged: 12 patients showed an increase in CD34+ cells in PB during the first 4 h after G-CSF (POM I), while eight patients had an initial decrease of CD34+ cells. However, all patients together showed a significant increase of CD34+ cells about 10 h after G-CSF administration. Further studies with more patients, using an enhanced monitoring schedule will be required to refine the results. PMID- 16044143 TI - Transfusion of leukoreduced cellular blood components from cytomegalovirus unscreened donors in allogeneic hematopoietic transplant recipients: analysis of 72 recipients. AB - Leukoreduction of blood components has been considered a safe alternative to screening donors for CMV. The objective of this study is to analyze the effectiveness of bedside leukoreduction in preventing CMV transmission. We retrospectively studied 72 transplant recipients and donors who were CMV seronegative pairs. All patients were transfused with CMV-unscreened cellular blood products leukoreduced at the bedside using leukoreduction filters. Quality control measures performed monthly in our leukoreduced blood components consistently demonstrated that at least 95% of the units sampled meet the leukoreduction criterion established by the American Association of Blood Banks standards. The CMV status of the recipients and donors was determined before transplantation by the latex agglutination assay. Recipients were observed for at least 100 days after transplantation. CMV cultures of urine, buffy coat, bone marrow, and bronchial washings were done weekly when indicated. CMV antigenemia testing was performed twice weekly: 11 transplant recipients seroconverted after transplantation. One patient was positive for CMV antigenemia 4 months after transplantation, but did not have CMV infection. Two of 61 patients who were not seroconverted were CMV antigenemia positive and did not have CMV infection: leukoreduction of cellular blood products is an efficient method of preventing CMV infection. PMID- 16044144 TI - Economic analysis of unrelated allogeneic bone marrow transplantation: results from the randomized clinical trial of T-cell depletion vs unmanipulated grafts for the prevention of graft-versus-host disease. AB - Unrelated-donor marrow transplantation is a potential option for transplant candidates lacking a compatible related donor. The T-cell Depletion Study compared the 3-year disease-free survival for patients receiving T-cell-depleted (TCD) donor marrow (n = 203) vs unmanipulated donor marrow with methotrexate and cyclosporine (M/C) (n = 207). Hospital costs during index admission were documented with billing data, while hospital costs during subsequent 6-month follow-up were estimated from case report forms. Patients with index admission billing were included in the analysis (TCD = 119, M/C = 127). Total hospital length of stay (LOS) was similar across groups, with medians 47.0 days for TCD and 52.0 days for M/C (P = 0.72). Total hospital costs were comparable, 145,115 dollars vs 141,981 dollars (P = 0.63) for TCD and M/C, respectively. However, controlling for site and patient characteristics, TCD was associated with a 12.1% reduction in LOS for the index admission (95% CI -19.4%, -4.3%). Independent of treatment, HLA matching (6/6) was associated with an 8.6% (95% CI -17.4%, +1.2%) reduction in the index admission LOS, while cost was lower by 15.8% (95% CI 26.7%, -3.3%). Treatment costs were similar for TCD and M/C study groups. Savings on reduced cost for treating acute graft-versus-host disease were likely offset by increase in serious infections in the TCD arm. PMID- 16044146 TI - A prognostic model for prolonged event-free survival after autologous or allogeneic blood or marrow transplantation for relapsed and refractory Hodgkin's disease. PMID- 16044147 TI - TP53INP1 is a novel p73 target gene that induces cell cycle arrest and cell death by modulating p73 transcriptional activity. AB - TP53INP1 is an alternatively spliced gene encoding two nuclear protein isoforms (TP53INP1alpha and TP53INP1beta), whose transcription is activated by p53. When overexpressed, both isoforms induce cell cycle arrest in G1 and enhance p53 mediated apoptosis. TP53INP1s also interact with the p53 gene and regulate p53 transcriptional activity. We report here that TP53INP1 expression is induced during experimental acute pancreatitis in p53-/- mice and in cisplatin-treated p53-/- mouse embryo fibroblasts (MEFs). We demonstrate that ectopic expression of p73, a p53 homologue, leads to TP53INP1 induction in p53-deficient cells. In turn, TP53INP1s alters the transactivation capacity of p73 on several p53-target genes, including TP53INP1 itself, demonstrating a functional association between p73 and TP53INP1s. Also, when overexpressed in p53-deficient cells, TP53INP1s inhibit cell growth and promote cell death as assessed by cell cycle analysis and colony formation assays. Finally, we show that TP53INP1s potentiate the capacity of p73 to inhibit cell growth, that effect being prevented when the p53 mutant R175H is expressed or when p73 expression is blocked by a siRNA. These results suggest that TP53INP1s are functionally associated with p73 to regulate cell cycle progression and apoptosis, independently from p53. PMID- 16044148 TI - Loss of TNF-alpha-regulated COX-2 expression in ovarian cancer cells. AB - Cyclooxygenase 2 (COX-2) is often found overexpressed in cancer and is thought to have a role in carcinogenic promotion, and thus is a target for therapeutic intervention. Here, we investigated the regulation of COX-2 expression in normal and cancer ovarian surface epithelial cells. Tumor necrosis factor alpha (TNF alpha) is a potent inducer of COX-2 expression in the ovarian surface epithelium and this regulation is a critical step in ovulation. We observed that TNF-alpha stimulated COX-2 expression in human primary and immortalized epithelial (HIO) cell lines. The stimulation was suppressed by inhibitors of several signaling pathways, indicating the collaboration of TNF-alpha-activated signaling pathways mediates the regulation of COX-2 expression. In five ovarian cancer cell lines analysed, four did not express detectable COX-2 and TNF-alpha failed to elicit COX-2 expression. In NIH:OVCAR-5, the only ovarian cancer cell line expressing COX-2, signal pathway inhibitors no longer affected TNF-alpha-induced COX-2 expression. Thus, we conclude that TNF-alpha mediated signaling is uncoupled from the modulation of COX-2 expression in ovarian cancer. The loss of COX-2 expression was also observed to associate closely with epithelial neoplastic morphological transformation. The frequent loss of COX-2 expression suggests in ovarian cancer, unlike in other epithelial cancers, COX-2 expression does not contribute to ovarian cancer malignancy. PMID- 16044149 TI - Activation of the protein kinase B pathway by the HPV-16 E7 oncoprotein occurs through a mechanism involving interaction with PP2A. AB - Protein kinase B (PKB) or Akt is one of several second messenger kinases that are activated by cell attachment and growth factor signaling, and that transmit signals to the cell nucleus to inhibit apoptosis and thereby increase cell survival during proliferation. Other viral proteins target this pathway by increasing PKB/Akt phosphorylation, and this pathway has been implicated in the transformation of human keratinocytes by HPV E6 and E7, together with activated notch 1. Here, we examine how HPV E7 expression affects the phosphorylation of PKB. We show that HPV-16 E7 increases the level of phosphorylation of PKB in response to serum stimulation, by a mechanism independent of downregulation of PTEN phosphatase, a known inhibitor of the PI3K (PI3 kinase) pathway. The use of specific antibodies shows that some proportion of PKB/Akt that is phosphorylated both on threonine 308 and serine 473 is maintained in the presence of E7 in a PI3 kinase-independent manner, and is activated for phosphorylation of BAD, a known downstream target of PKB/Akt. Use of E7 mutants has ruled out both an inhibition of IGFBP-3, a known E7 target and PKB/Akt modulator, and the interaction of E7 with cellular pocket proteins, as being the mechanism for the PKB/Akt stimulation. PKB binds PP2A and is a known substrate of PP2A. Here, we show that HPV E7 also binds to both the 35 kDa catalytic and 65 kDa structural subunits of PP2A, an interaction that sequesters these subunits and inhibits their interaction with PKB, thereby maintaining PKB/Akt signaling by inhibiting its dephosphorylation. PMID- 16044150 TI - Defining the oncogenic function of the TEL/AML1 (ETV6/RUNX1) fusion protein in a mouse model. AB - The t(12;21) translocation, generating the TEL/AML1 fusion protein, is the most common genetic lesion in childhood cancer. Using a bone marrow transplantation model, we demonstrate that TEL/AML1 expression impinges on normal hematopoietic differentiation, leading to the in vivo accumulation and persistence of an early progenitor compartment with a Sca1(+)/Kit(hi)/CD11b(+) phenotype and an increased self-renewal capacity, as documented by replating assays in vitro. Differentiation of these cells is not blocked, but the frequency of mature blood cells arising from TEL/AML1-transduced progenitors is low. Impaired differentiation is prominently observed in the pro-B-cell compartment, resulting in an proportional increase in early progenitors in vivo, consistent with the t(12;21) ALL phenotype. Despite the accumulation of both multipotent and B-cell progenitors in vivo, no leukemia induction was observed during an observation period of over 1 year. These results are consistent with findings in twins with concordant ALL, showing that TEL/AML1 generates a preleukemic clone in utero that persists for several years in a clinically covert fashion. Furthermore, our studies showed that the pointed domain of TEL/AML1, which recruits transcriptional repressors and directs oligomerization with either TEL/AML1 or wild-type TEL, was essential for the observed differentiation impairment and could not be replaced with another oligomerization domain. PMID- 16044151 TI - Unique aspects of mda-7/IL-24 antitumor bystander activity: establishing a role for secretion of MDA-7/IL-24 protein by normal cells. AB - Melanoma differentiation associated gene-7 (mda-7) was cloned using subtraction hybridization from terminally differentiated human melanoma cells. Based on structural and functional properties, mda-7 is now recognized as interleukin-24 (IL-24), a new member of the expanding IL-10 gene family. Unique properties of mda-7/IL-24 include its ability to selectively induce growth suppression, apoptosis and radiosensitization in diverse human cancer cells, without causing similar effects in normal cells. The utility of mda-7/IL-24, administered by means of a replication-incompetent adenovirus, as a gene therapy for cancer has recently received validation in patients, highlighting an important phenomenon initially observed in pancreatic tumor cells, namely a 'potent bystander apoptosis-inducing effect' in adjacent tumor cells not initially receiving this gene product. We presently investigated the contribution of mda-7/IL-24 secreted by normal cells in mediating this 'bystander effect', and document that normal cells induced to produce mda-7/IL-24 following infection with recombinant adenoviruses expressing this cytokine secrete mda-7/IL-24, which modifies the anchorage-independent growth, invasiveness, survival and sensitivity to radiation of cancer cells that contain functional IL-20/IL-22 receptors, but not in cancer cells that lack a complete set of receptors. Moreover, the combination of secreted mda-7/IL-24 and radiation engenders a 'bystander antitumor effect' not only in inherently mda-7/IL-24 or radiation-sensitive cancer cells, but also in tumor cells overexpressing the antiapoptotic proteins bcl-2 or bcl-x(L) and displaying resistance to either treatment alone. The present studies provide definitive evidence that secreted mda-7/IL-24 from normal cells can induce direct antitumor and radiation-enhancing effects that are dependent on the presence of canonical receptors for this cytokine on tumor cells. Moreover, we now describe a novel means of enhancing mda-7/IL-24's therapeutic potential by targeting normal cells to produce and release this cancer-specific apoptosis-inducing cytokine, a strategy that could be employed as an innovative way of using this unique gene product for treating metastatic disease. PMID- 16044152 TI - Prediction of doxorubicin sensitivity in breast tumors based on gene expression profiles of drug-resistant cell lines correlates with patient survival. AB - Up to date clinical tests for predicting cancer chemotherapy response are not available and individual markers have shown little predictive value. We hypothesized that gene expression patterns attributable to chemotherapy-resistant cells can predict response and cancer prognosis. We contrasted the expression profiles of 13 different human tumor cell lines of gastric (EPG85-257), pancreatic (EPP85-181), colon (HT29) and breast (MCF7 and MDA-MB-231) origin and their counterparts resistant to the topoisomerase inhibitors daunorubicin, doxorubicin or mitoxantrone. We interrogated cDNA arrays with 43 000 cDNA clones ( approximately 30 000 unique genes) to study the expression pattern of these cell lines. We divided gene expression profiles into two sets: we compared the expression patterns of the daunorubicin/doxorubicin-resistant cell lines and the mitoxantrone-resistant cell lines independently to the parental cell lines. For identifying predictive genes, the Prediction Analysis for Mircorarrays algorithm was used. The analysis revealed 79 genes best correlated with doxorubicin resistance and 70 genes with mitoxantrone resistance. In an independent classification experiment, we applied our model of resistance for predicting the sensitivity of 44 previously characterized breast cancer samples. The patient group characterized by the gene expression profile similar to those of doxorubicin-sensitive cell lines exhibited longer survival (49.7+/-26.1 months, n=21, P=0.034) than the resistant group (32.9+/-18.7 months, n=23). The application of gene expression signatures derived from doxorubicin-resistant and sensitive cell lines allowed to predict effectively clinical survival after doxorubicin monotherapy. Our approach demonstrates the significance of in vitro experiments in the development of new strategies for cancer response prediction. PMID- 16044153 TI - MEK kinase 1 is essential for Bcr-Abl-induced STAT3 and self-renewal activity in embryonic stem cells. AB - BCR-ABL oncogene, the molecular hallmark of chronic myelogenous leukemia, arises in a primitive hematopoietic stem cell that has the capacity for both differentiation and self-renewal. Its product, Bcr-Abl protein, has been shown to activate signal transducers and activators of transcription 3 (STAT3) and to promote self-renewal in embryonic stem (ES) cells, even in the absence of leukemia inhibitory factor (LIF). MEK kinase 1 (MEKK1) is a 196-kDa mitogen activated protein kinase (MAPK) kinase kinase involved in Bcr-Abl signal transduction. To investigate the role of MEKK1 in Bcr-Abl-induced transformation of stem cells, p210 Bcr-Abl was stably transfected into wild-type (WT(p210)) and MEKK1-/- (MEKK1-/-(p210)) ES cells. Bcr-Abl enhanced MEKK1 expression in ES transfectants, as it does in other Bcr-Abl-transformed cells. In the absence of LIF, WT(p210) cells showed constitutive STAT3 activation and formed rounded, compact colonies having strong alkaline phosphatase activity, a characteristic phenotype of undifferentiated ES cells. MEKK1-/-(p210) cells, by contrast, showed less STAT3 activity than WT(p210) cells and formed large, flattened colonies having weak alkaline phosphatase activity, a phenotype of differentiated ES cells. These results indicate that MEKK1 plays a key role in Bcr-Abl-induced STAT3 activation and in ES cells' capacity for LIF-independent self-renewal, and may thus be involved in Bcr-Abl-mediated leukemogenesis in stem cells. PMID- 16044154 TI - Retinoid-induced activation of NF-kappaB in APL cells is not essential for granulocytic differentiation, but prolongs the life span of mature cells. AB - All-trans retinoic acid (ATRA) significantly improves the survival of patients with acute promyelocytic leukemia (APL) by inducing granulocytic differentiation of leukemia cells. Since an activation of the transcription factor NF-kappaB occurs during ATRA-induced maturation of APL cells, a mechanistic link between these two processes was investigated. Using an in vitro model for APL, we report that ectopic overexpression of a repressor of NF-kappaB activation did not affect granulocytic differentiation. Importantly, NF-kappaB inhibition markedly resulted in a decreased viability of the differentiated cells, which correlated with increased apoptosis. Apoptosis was accompanied by a sustained activation of the c Jun N-terminal kinase (JNK). Inhibition of JNK by the specific inhibitor SP600125 or by transfection of a dominant-negative mutant of JNK1 reduced the percentage of apoptotic cells, thus showing that JNK activation constitutes a death signal. Furthermore, impairment of NF-kappaB activation resulted in increased levels of reactive oxygen species (ROS) upon ATRA treatment. ROS accumulation was suppressed by the antioxidant butylated hydroxyanisol, which also abolished ATRA induced JNK activation and apoptosis. Altogether, our results demonstrate an anti apoptotic effect of NF-kappaB activation during ATRA-induced differentiation of NB4 cells and identify repression of ROS-mediated JNK activation as a mechanism for this effect. Our observations also suggest that NF-kappaB signalling may contribute to an accumulation of mature APL cells and participate in the development of ATRA syndrome. PMID- 16044155 TI - A small molecule Smac-mimic compound induces apoptosis and sensitizes TRAIL- and etoposide-induced apoptosis in breast cancer cells. AB - Inhibitor of apoptosis protein (IAP) suppresses apoptosis through binding and inhibiting active caspases-3, -7 and -9 via its baculoviral IAP repeat (BIR) domains. During apoptosis the caspase inhibition by IAPs can be negatively regulated by a mitochondrial protein second mitochondrial-derived activator of caspase (Smac). Smac physically interacts with multiple IAPs and relieves their inhibitory effect on caspases-3, -7 and -9. Recently, a small molecule Smac-mimic compound (Smac-mimic), which potentiates TNF-related apoptosis-inducing ligand (TRAIL) and tumor necrosis factor (TNF)-alpha mediated cell death in glioblastoma T98G cells and HeLa cells, was identified and characterized. To determine the efficacy of this compound in breast cancer cells, we first measured protein expression of three IAPs: XIAP, cIAP-1, and cIAP-2 in nine independent breast cancer cell lines. Three cell lines were chosen: a high IAPs expressing line MDA MB-231, and two low IAPs expressing lines, T47D and MDA-MB-453. The cell lines were tested for their sensitivity to Smac-mimic alone or in combination with TRAIL or etoposide. Acting alone, Smac-mimic was quite potent with a cytotoxic IC50 of 3.8 nM in high IAPs expressing MDA-MB-231 cells, but was inactive at a much higher concentration in low IAPs expressing T47D and MDA-MB-453 cells. In fact, as low as 2.5 nM of Smac-mimic alone was sufficient to activate caspase-3 and induce apoptosis in MDA-MB-231 cells. In combinational treatments with TRAIL or etoposide, Smac-mimic significantly sensitized cells to growth suppression in MDA-MB-231 cells, but to a lesser extent in T47D and MDA-MB-453 cells. Furthermore, it significantly synergized MDA-MB-231, but not T47D cells to apoptosis induced by either TRAIL or etoposide. Thus, in these cell lines, Smac mimic acts in an apparent IAPs dependent manner to induce apoptosis alone as well as sensitizes breast cancer cells to TRAIL or etoposide induced apoptosis via caspase-3 activation. PMID- 16044156 TI - Tumor-specific exon 1 mutations could be the 'hit event' predisposing Rb2/p130 gene to epigenetic silencing in lung cancer. AB - Genetic alterations in Rb2/p130 gene have been reported in several tumors, but till now there are insufficient and conflicting data linking the loss of pRb2/p130 expression with the mutational status of this gene in lung cancer. We recently reported that loss or lowering of pRb2/p130 expression is mainly due to aberrant Rb2/p130 promoter methylation, in retinoblastoma tumors, and indicated that epigenetic silencing of Rb2/p130 can impair its function to negatively regulate cell cycle progression as well as apoptotic response. In order to clarify Rb2/p130 gene inactivation in lung cancer, we investigated whether epigenetic events could impair the expression of this gene in NSLC. Here, we show that specific Rb2-exon 1 homozygous mutations, occurring in an Rb2/p130, region, rich in CpG dinucleotides, could be the 'hit event' that predispose this gene to epigenetic changes, leading to Rb2/p130 gene silencing in lung cancer. Moreover, these homozygous mutations, found in different tumor histotypes, could represent tumor-specific markers. PMID- 16044157 TI - Inhibition of histone deacetylase 3 stimulates apoptosis induced by heat shock under acidic conditions in human maxillary cancer. AB - To elucidate the molecular mechanisms for the enhancement of heat-induced apoptosis on exposure to acidic conditions, human maxillary carcinoma IMC-3 cells were heat-shocked at 44 degrees C for 30 min at either pH 7.4 or 6.7. Analyses with cDNA arrays, the reverse transcription-polymerase chain reaction (RT-PCR), and Western blotting were performed. We found that histone deacetylase 3 (HDAC3) was specifically induced after hyperthermia at 44 degrees C for 30 min at pH 6.7. Although the cytotoxicity of heating at 44 degrees C for 30 min was enhanced by decreasing the pH from 7.4 to 6.7, it was enhanced even more by antisense RNA oligonucleotides for HDAC3. The induction of G2/M arrest after heating occurred earlier at pH 6.7 than at pH 7.4. The inhibition of HDAC3 by the antisense RNA oligonucleotides suppressed partially the induction of G2/M arrest, resulting in an enhancement of the apoptosis caused by the heating under acidic conditions. Antisense RNA oligonucleotides for HDAC3 enhanced apoptosis 48 h after hyperthermia at 43 degrees C for 30 min in vivo. Analyses of p65 activity suggested that NF-kappaB is involved in this enhancement of hyperthermia. HDAC3 may be a novel target enhancing hyperthermia and combined treatment with hyperthermia and HDAC inhibitors is a possible modality for cancer therapy. PMID- 16044158 TI - DNA damage in transcribed genes induces apoptosis via the JNK pathway and the JNK phosphatase MKP-1. AB - The nucleotide excision repair (NER) system consists of two sub-pathways, global genome repair (GGR) and transcription-coupled repair (TCR), which exhibit distinct functions in the cellular response to genotoxic stress. Defects in TCR result in prolonged UV light-induced stalling of RNA polymerase II and hypersensitivity to apoptosis induced by UV and certain chemotherapeutic drugs. Here, we show that low doses of UV trigger delayed activation of the stress induced MAPkinase JNK and its proapoptotic targets c-Jun and ATF-3 in TCR deficient primary human fibroblasts from Xeroderma Pigmentosum (XP) and Cockayne syndrome (CS) patients. This delayed activation of the JNK pathway is not observed in GGR-deficient TCR-proficient XP cells, is independent of functional p53, and is established through repression of the JNK-phosphatase MKP-1 rather than by activation of the JNK kinases MKK4 and 7. Enzymatic reversal of UV induced cyclobutane pyrimidine dimers (CPDs) by CPD photolyase abrogated JNK activation, MKP-1 repression, and apoptosis in TCR-deficient XPA cells. Ectopic expression of MKP-1 inhibited DNA-damage-induced JNK activity and apoptosis. These results identify both MKP-1 and JNK as sensors and downstream effectors of persistent DNA damage in transcribed genes and suggest a link between the JNK pathway and UV-induced stalling of RNApol II. PMID- 16044159 TI - Raised expression of the antiapoptotic protein ped/pea-15 increases susceptibility to chemically induced skin tumor development. AB - ped/pea-15 is a cytosolic protein performing a broad antiapoptotic function. We show that, upon DMBA/TPA-induced skin carcinogenesis, transgenic mice overexpressing ped/pea-15 (Tg(ped/pea-15)) display early development of papillomas and a four-fold increase in papilloma number compared to the nontransgenic littermates (P<0.001). The malignant conversion frequency was 24% for the Tg(ped/pea-15) mice and only 5% in controls (P<0.01). The isolated application of TPA, but not that of DMBA, was sufficient to reversibly upregulate ped/pea-15 in both untransformed skin and cultured keratinocytes. ped/pea-15 protein levels were also increased in DMBA/TPA-induced papillomas of both Tg(ped/pea-15) and control mice. Isolated TPA applications induced Caspase-3 activation and apoptosis in nontransformed mouse epidermal tissues. The induction of both Caspase-3 and apoptosis by TPA were four-fold inhibited in the skin of the Tg(ped/pea-15) compared to the nontransgenic mice, accompanied by a similarly sized reduction in TPA-induced JNK and p38 stimulation and by constitutive induction of cytoplasmic ERK activity in the transgenics. ped/pea-15 expression was stably increased in cell lines from DMBA/TPA-induced skin papillomas and carcinomas, paralleled by protection from TPA apoptosis. In the A5 spindle carcinoma cell line, antisense inhibition of ped/pea-15 expression simultaneously rescued sensitivity to TPA-induced Caspase-3 function and apoptosis. The antisense also reduced A5 cell ability to grow in semisolid media by 65% (P<0.001) and increased by three-fold tumor latency time (P<0.01). Thus, the expression levels of ped/pea-15 control Caspase-3 function and epidermal cell apoptosis in vivo and determine susceptibility to skin tumor development. PMID- 16044160 TI - Absence of mutation in the putative tumor-suppressor gene KLF6 in colorectal cancers. AB - The KLF6 gene encodes the Kruppel-like factor 6, a transcription factor that has been individualized as a tumor-suppressor gene involved in the regulation of cell proliferation and differentiation. Recently, high frequency (42%) of KLF6 mutations have been reported in colorectal cancers (CRC) as in prostate cancers, astrocytic gliomas and hepatocellular carcinomas. The aims of the study was to confirm the frequency of KLF6 mutations in a larger series of CRC than that previously published by using DNA extracted from frozen tissue samples, which have been proved to generate less mutational artefact than that extracted from formalin-fixed paraffin-embedded tissue samples, in order to compare KLF6 mutation frequency with that of other common genetic alterations and to determine genotype-phenotype correlations. Amplification and direct sequencing of KLF6 exon 2 of 76 CRC and matched normal frozen tissues was performed. Polymorphisms were observed in 14 cases, among which two (T35T and S116S) had not already been reported. No KLF6 somatic mutation was observed. Our data suggest a minor role of KLF6 mutation in colorectal carcinogenesis and underline the fact that the validity of sequence informations obtained from DNA extracted from formalin-fixed tissues may be limited. PMID- 16044161 TI - Curcumin suppresses growth and induces apoptosis in primary effusion lymphoma. AB - The mechanisms that regulate induction of the antiapoptotic state and mitogenic signals in primary effusion lymphoma (PEL) are not well known. In efforts to identify novel approaches to block the proliferation of PEL cells, we found that curcumin (diferuloylmethane), a natural compound isolated from the plant Curcuma Ionga, inhibits cell proliferation and induces apoptosis in a dose dependent manner in several PEL cell lines. Such effects of curcumin appear to result from suppression of the constitutively active STAT3 through inhibition of Janus kinase 1 (JAK1). Our data also demonstrate that curcumin induces loss of mitochondrial membrane potential with subsequent release of cytochrome c and activation of caspase-3, followed by polyadenosin-5'-diphosphate-ribose polymerase (PARP) cleavage. Altogether, our findings suggest a novel function for curcumin, acting as a suppressor of JAK-1 and STAT3 activation in PEL cells, leading to inhibition of proliferation and induction of caspase-dependent apoptosis. Therefore, curcumin may have a future therapeutic role in PEL and possibly other malignancies with constitutive activation of STAT3. PMID- 16044162 TI - Lysophosphatidic acid and endothelin-induced proliferation of ovarian cancer cell lines is mitigated by neutralization of granulin-epithelin precursor (GEP), a prosurvival factor for ovarian cancer. AB - Granulin-epithelin precursor (GEP/progranulin) is an autocrine growth factor for ovarian cancer. We examined the production and function of GEP and report that: (1) GEP production is regulated by endothelin (ET-1), lysophosphatidic acid (LPA), and cAMP; (2) cAMP signals GEP production through exchange protein activated by cAMP (EPAC); (3) ET-1 and cAMP/EPAC induce GEP through ERK1/2; and (4) neutralization of GEP results in apoptosis. Exposure of HEY-A8 and OVCAR3 ovarian cancer cells to LPA and ET-1 yielded GEP production and secretion in a dose- and time-dependent fashion; neither stimulated significant concentrations of cAMP directly. Stimulation of cAMP production with pertussis and cholera toxin, or forskolin induced GEP in a PKA-independent fashion. EPAC, an intracellular cAMP receptor, is activated specifically by the cAMP analog, 8-CPT 2'-O-Me-cAMP (8-CPT); 8-CPT treatment stimulated GEP production and secretion. The MEK inhibitor, U0126, abrogated GEP production in response to ET-1 and 8-CPT, confirming involvement of MAPK. A partial inhibition of basal and stimulated GEP production was observed when cells were treated with a internal calcium chelator, BAPTA. Neutralizing anti-GEP antibody reversed basal as well as LPA, ET-1 and 8 CPT-induced ovarian cancer cell growth and induced apoptosis as demonstrated by caspase-3 and PARP cleavage, DNA fragmentation, and nuclear condensation. These results indicate that GEP is a growth and survival factor for ovarian cancer, induced by LPA and ET-1 and cAMP/EPAC through ERK1/2. PMID- 16044163 TI - ARK5 is transcriptionally regulated by the Large-MAF family and mediates IGF-1 induced cell invasion in multiple myeloma: ARK5 as a new molecular determinant of malignant multiple myeloma. AB - ARK5, AMP-activated protein kinase (AMPK)-related protein kinase mediating Akt signals, is closely involved in tumor progression, and its stage-associated expression was observed in colorectal cancer. In this study, we found ARK5 expression in multiple myeloma cell lines expressing c-MAF and MAFB. In addition, gene expression profiling of 351 clinical specimens revealed ARK5 expression in primary myelomas expressing c-MAF and MAFB, suggesting that ARK5 may be a transcriptional target of the Large-MAF family. Sequence analysis of the ARK5 gene promoter revealed that it contains two putative MAF-recognition element (MARE) sequences. In support of this hypothesis, ARK5 was induced when an MAFB or c-MAF expression vector was introduced into non-ARK5-expressing colon cancer cells. Furthermore, ARK5 promoter activity was dramatically decreased by mutation or deletion of MARE sequences. Chromatin immunoprecipitation assays revealed an interaction between the Large-MAF family proteins and MARE sequences in the ARK5 promoter. Moreover, in ARK5 mRNA-expressing multiple myeloma lines, but not in ARK5-negative lines, insulin-like growth factor (IGF)-1 increased invasion activity. IGF-1-induced invasion was reproduced when ARK5 was overexpressed in Burkitt's lymphoma and plasmacytoma lines. Based on results, we conclude that ARK5 is a transcriptional target of the Large-MAF family through MARE sequence and that ARK5 may in part mediate the aggressive phenotype associated with c-MAF- and MAFB-expressing myelomas. PMID- 16044164 TI - Distinct CpG methylation profiles characterize different clinical groups of neuroblastic tumors. AB - The hypermethylation of CpG islands within gene promoter regions is an epigenetic phenomenon that is often, but not always, associated with the transcriptional silencing of downstream genes and contributes to carcinogenesis. We have determined the pattern of methylation of several genes involved in distinct biological pathways, including cell proliferation and apoptosis, in neuroblastoma and in the nonmalignant ganglioneuroma. The purpose of this work was to search for epigenetic signatures that could be associated with defined clinical and biological parameters and that, in prospective, could identify specific risk categories among the patients. We have analysed 31 malignant neuroblastoma with or without MYCN amplification and 13 benign ganglioneuroma and we have observed dramatic differences in the methylation pattern of five genes (CASP8, 14.3.3sigma, DeltaN-p73, RASSF1A and DCR2) between these tumors indicating that this phenomenon is not tissue-specific and can be considered as cancer-dependent. Furthermore, the methylation pattern of 14.3.3sigma, RASSF1A and of an intragenic segment of CASP8 was significantly different between MYCN amplified and single copy neuroblastoma suggesting a specific role of epigenetic alterations in aggressive neuroblastoma. PMID- 16044165 TI - Pharmacologic profiling of transcriptional targets deciphers promoter logic. AB - The blueprint for cellular diversity and response to environmental change is encoded in the cis-acting regulatory sequences of most genes. Deciphering this 'cis-regulatory code' requires multivariate data sets that examine how these regions coordinate transcription in response to diverse environmental stimuli and therapeutic treatments. We describe a transcriptional approach that profiles the activation of multiple transcriptional targets against combinatorial arrays of therapeutic and signal transducing agents. Application of this approach demonstrates how cis-element composition and promoter context combine to influence transcription downstream of mitogen-induced signaling networks. Computational dissection of these transcriptional profiles in activated T cells uncovers a novel regulatory synergy between IGF-1 and CD28 costimulation that modulates NF-kappaB and AP1 pathways through signaling cascades sensitive to cyclosporin A and wortmannin. This approach provides a broader view of the hierarchical signal integration governing gene expression and will facilitate a practical design of combinatorial therapeutic strategies for exploiting critical control points in transcriptional regulation. PMID- 16044166 TI - The experimental study of hypoxia-inducible factor-1alpha and its target genes in spinal cord injury. AB - STUDY DESIGN: Animal model of compressive spinal cord injury (SCI), reverse transcription-polymerase chain reaction (RT-PCR), in situ hybridization (ISH), immunohistochemistry (IHC) and enzymehistochemistry (EHC) were used to test the hypothesis that hypoxia-inducible factor-1alpha (HIF-1alpha) and the target genes activated by HIF-1alpha are involved in cell hypoxia tolerance and tissue vascularity to help injured tissue to go through the stress disease. OBJECTIVE: To determine whether HIF-1alpha and its target genes associated with hypoxia tolerance and neovascularization take part in the pathophysiological procedure of SCI in rats. SETTING: Yunnan University, China. METHODS: Random-bred adult male Sprague-Dawley (SD) rats weighing 250+/-50 g were prepared for compressive SCI models. After receiving compressive injury at T(10), rats were sacrificed at different times from 6 h to 1 week after injury. The injured cords were removed, and HIF-1alpha and its target genes were assayed by RT-PCR, ISH, IHC and EHC. The data were statistically analyzed. RESULTS: An increase in HIF-1alpha mRNA expression was observed 12 h postinjury, reached a maximum at 3 days, and reduced gradually thereafter. HIF-1alpha protein expressed earlier than HIF-1alpha mRNA. Additionally, two glycolytic enzymes and vascular endothelial growth factor (VEGF), which are regulated by HIF-1alpha, also increased after an interval postinjury, and their expression patterns shared a same trend with that of HIF 1alpha protein. CONCLUSION: The findings suggested that the most important hypoxic regulatory factor HIF-1alpha was upregulated in involved cells by activating the transcription and increasing protein stability, and subsequently activated the expression of HIF-1alpha target genes, including glycolytic enzymes and VEGF in SCI. Combined with the pathologic observation, it suggested that overexpression of HIF-1alpha and its target genes might take part in hypoxia tolerance and vascularity of the injured spinal cord. PMID- 16044167 TI - Continent cutaneous urinary diversion in patients with spinal cord injury. AB - STUDY DESIGN: Retrospective study. OBJECTIVE: To examine the functional results and effect on quality of life of continent cutaneous urinary diversion in spinal cord injured patients. SETTING: Department of Urology, Sahlgrenska University Hospital, Goteborg, Sweden. SUBJECTS: A total of 10 patients with spinal cord injury (SCI). METHOD: The patients were operated on with an ileal reservoir (Kock reservoir or T-pouch), Cr-EDTA clearance was determined preoperatively and at follow-up. The patients answered a questionnaire concerning reservoir function, various activities and quality of life. The patient charts were reviewed. RESULTS: One patient died of pulmonary embolism 3 years after surgery. Two patients were reoperated on for reservoir perforation. All patients were satisfied/very satisfied with their reservoirs. Half of them reported improved ability to perform various activities. Eight out of nine patients reported improved quality of life. CONCLUSION: For a selected group of patients with SCI, continent cutaneous urinary diversion provides successful outcome with improved quality of life. PMID- 16044168 TI - Restoration of frequency-dependent depression of the H-reflex by passive exercise in spinal rats. AB - STUDY DESIGN: Hyper-reflexia, measured as a decrease of low frequency-dependent depression of the H-reflex, is known to occur in both humans and animals after spinal cord injury (SCI). Previous studies have shown that passive exercise for 3 months could be used to restore low frequency-dependent depression of the H reflex after SCI. OBJECTIVE: To determine the effects of various periods of time on the ability of passive exercise to restore low frequency-dependent depression of the H-reflex. SETTING: Spinal Cord Injury Mobilization Program of the Center for Translational Neuroscience, the research arm of the Jackson T Stephens Spine and Neuroscience Institute, Little Rock, AR, USA. METHODS: Adult rats underwent complete spinal cord transection at the T10 level. The hindlimbs were passively exercised in different groups of rats for 1 h/day, 5 days/week for 15, 30, 45, 60, or 90 days, and low frequency-dependent depression of the H-reflex was tested. RESULTS: Statistically significant low frequency-dependent depression of the H-reflex was evident by 30 days of exercise, although numerical reductions were seen even at 15 days. There was a linear decrease in low frequency-dependent depression of the H-reflex with duration of passive exercise. CONCLUSIONS: Passive exercise can restore frequency-dependent depression of spinal reflexes in a time-dependent manner if used following complete spinal transection. PMID- 16044169 TI - Memokath stents for the treatment of detrusor sphincter dyssynergia (DSD) in men with spinal cord injury: the Princess Royal Spinal Injuries Unit 10-year experience. AB - STUDY DESIGN: Medical records review. OBJECTIVE: To assess the effectiveness of the Memokath (Engineers and Doctors A/S, Denmark) thermosensitive stent as a 'nondestructive' means of reducing bladder outlet resistance by treating detrusor sphincter dyssynergia (DSD) of neurogenic bladder dysfunction associated with spinal cord injury. SETTING: Spinal Injuries Unit, Sheffield, England. METHODS: A medical records review was performed to examine our experience of Memokaths over the last 10 years. During this time, 29 patients with spinal cord injury (17 tetraplegic and 12 paraplegic) underwent stenting of the external urethral sphincter either for prevention of dysreflexic symptoms, high residual urine volumes and subsequent urinary tract infection (UTI) or for protection of the upper tracts. RESULTS: A total of 33 stents were inserted into 29 men (25-77 years) with suprasacral spinal cord injury. Initial results showed that the Memokath was effective in almost all for relief of dysreflexic symptoms and elimination of DSD on pressure flow urodynamics. However, to date, 30 of the 33 stents have been removed. The overall mean working life of the Memokath was 21 months. Four stents were removed electively and 23 for complications, which included stent migration (seven) and blockage (14). Single-ended stents were more prone to migration, which was rare after 1 year (1-13 months, median 3 months, mean 5.5 months). Stent blockage by encrustation or prostatic ingrowth did not occur before 12 months (12-45 months, median 30, mean 27.9 months). CONCLUSIONS: In selected patients, temporary, thermo-expandable (Memokath) stents are effective in the treatment of DSD. The 'working life' of a Memokath stent is 21 months; however, complications do occur which may necessitate removal. Our overall experience with Memokath stents was disappointing. In future, Memokath stents will only be inserted after careful consideration in patients with prior 'failed' transurethral sphincterotomy or with caution in patients suitable for reconstructive surgery. PMID- 16044170 TI - Gene expression and association analyses of LIM (PDLIM5) in bipolar disorder and schizophrenia. AB - We previously reported that expression level of LIM (ENH, PDLIM5) was significantly and commonly increased in the brains of patients with bipolar disorder, schizophrenia, and major depression. Expression of LIM was decreased in the lymphoblastoid cells derived from patients with bipolar disorders and schizophrenia. LIM protein reportedly plays an important role in linking protein kinase C with calcium channel. These findings suggested the role of LIM in the pathophysiology of bipolar disorder and schizophrenia. To further investigate the role of LIM in these mental disorders, we performed a replication study of gene expression analysis and performed genetic association studies. Upregulation of LIM was confirmed in the independent sample set obtained from Stanley Array Collection. No effect of sample pH or medication was observed. Genetic association study revealed the association of single nucleotide polymorphism (SNP)1 (rs10008257) with bipolar disorder. In an independent sample set, SNP2 (rs2433320) close to SNP1 was associated with bipolar disorder. In total samples, haplotype of these two SNPs was associated with bipolar disorder. No association was observed in case-control analysis and family-based association analysis in schizophrenia. These results suggest that SNPs in the upstream region of LIM may confer the genetic risk for bipolar disorder. PMID- 16044171 TI - Identifying potential risk haplotypes for schizophrenia at the DTNBP1 locus in Han Chinese and Scottish populations. AB - The dystrobrevin-binding protein 1 (DTNBP1) gene on chromosome 6p has emerged as a potential susceptibility gene for schizophrenia. Although a number of attempts to replicate the original association finding have been successful, they have not identified any obvious pathogenic variants or a single at risk haplotype common to all populations studied. In the present study we attempted further replication in an independent sample of 638 nuclear families from the Han Chinese population of Sichuan Province, SW China. We also examined 580 Scottish schizophrenic cases and 620 controls. We genotyped 10 single-nucleotide polymorphisms (SNPs) in DTNBP1 that were used in the original report of association, plus rs2619538 (SNP 'A') in the putative promoter region, which has also been associated with schizophrenia. In the Chinese trios we found that two SNPs (P1635 and P1765) were significantly overtransmitted, but with alleles opposite to those reported in the original studies. SNPs P1757 and P1765 formed a common haplotype, which also showed significant overtransmission. In the Scottish cases and controls, no individual markers were significantly associated with schizophrenia. A single haplotype, which included rs2619538 and P1583, and one rare haplotype, composed of P1320 and P1757, were significantly associated with schizophrenia, but no previously reported haplotypes were associated. Based on the data from the Chinese population, our results provide statistical support for DTNBP1 as a susceptibility gene for schizophrenia, albeit with haplotypes different from those of the original study. However, our lack of replication in the Scottish samples also indicates that caution is warranted when evaluating the robustness of the evidence for DTNBP1 as genetic risk factor for schizophrenia. PMID- 16044172 TI - A regulatory variant of the human tryptophan hydroxylase-2 gene biases amygdala reactivity. AB - Recent studies have indicated that a newly identified second isoform of the tryptophan hydroxylase gene (TPH2) is preferentially involved in the rate limiting synthesis of neuronal serotonin. Genetic variation in the human TPH2 gene (hTPH2) has been associated with altered in vitro enzyme activity as well as increased risk for mood disorders. Here, we provide the first in vivo evidence that a relatively frequent regulatory variant (G(-844)T) of hTPH2 biases the reactivity of the amygdala, a neural structure critical in the generation and regulation of emotional behaviors. PMID- 16044173 TI - Alternative transcripts and evidence of imprinting of GNAL on 18p11.2. AB - Genetic studies implicating the region of human chromosome 18p11.2 in susceptibility to bipolar disorder and schizophrenia have observed parent-of origin effects that may be explained by genomic imprinting. We have identified a transcriptional variant of the GNAL gene in this region, employing an alternative first exon that is 5' to the originally identified start site. This alternative GNAL transcript encodes a longer functional variant of the stimulatory G-protein alpha subunit, Golf. The isoforms of Golf display different expression patterns in the CNS and functionally couple to the dopamine D1 receptor when heterologously expressed in Sf9 cells. In addition, there are CpG islands in the vicinity of both first exons that are differentially methylated, a hallmark of genomic imprinting. These results suggest that GNAL, and possibly other genes in the region, is subject to epigenetic regulation and strengthen the case for a susceptibility gene in this region. PMID- 16044174 TI - High amount of visceral fat mass is associated with multiple metabolic changes in offspring of type 2 diabetic patients. AB - OBJECTIVE: To investigate the relative contribution of total body fat mass (TFM) and intra-abdominal fat mass (IAFM) to metabolic consequences of obesity in offspring of type 2 diabetic parents. DESIGN: Cross-sectional study of 129 nondiabetic offspring of diabetic parents (59 men, 70 women, age 35.7 +/- 6.3 y, body mass index 26.2 +/- 4.6 kg/m2). Study subjects were grouped according to TFM (assessed with bioelectrical impedance) and IAFM (assessed with CT). Insulin sensitivity was assessed with the euglycemic hyperinsulinemic clamp, insulin secretion with the intravenous glucose tolerance test and energy expenditure with indirect calorimetry. Furthermore, C-reactive protein (CRP) and adiponectin levels were measured. RESULTS: Insulin resistance, low rates of oxidative and nonoxidative glucose disposal, high rates of lipid oxidation and reduced energy expenditure during hyperinsulinemia were associated with high IAFM, independently of TFM. Adiponectin level was reduced and CRP level increased in subjects with high IAFM. CONCLUSIONS: The metabolic changes relating to obesity are largely attributable to high IAFM, and are present even in normal weight subjects with high IAFM. PMID- 16044175 TI - Relationships between human serum resistin, inflammatory markers and insulin resistance. AB - OBJECTIVE: Data on the association of resistin levels with markers of insulin resistance are highly contrasting in humans and very few studies about its role in inflammation are available. This study investigates associations between serum resistin levels and markers of insulin resistance, inflammation (C-reactive protein (CRP)) and of oxidative stress (nytrotirosine (NT)). SUBJECTS: A randomly collected sample of 300 men from a population-based cohort was analysed, separated into two groups according to body mass index (BMI) and waist values. RESULTS: Correlations between resistin and BMI, waist, triglyceride, uric acid, fasting glucose, insulin and Homeostasis Model Assessment (HOMA) values were significant in subjects with normal BMI, but not in overweight/obese subjects. In a multiple regression model, after multiple adjustments and exclusion of diabetic patients, only fasting glucose remained significantly associated with resistin levels. Otherwise, resistin is associated to CRP levels in all individuals, after multiple adjustments and exclusion of diabetic patients (in normal BMI beta=0.82; 95% CI 0.21, 1.42; in overweight/obese beta=0.43; 95% CI 0.10, 0.76). In the same model, resistin values are negatively related to NT levels in normal weight individuals (beta=-1.61; 95% CI -0.77-2.45). CONCLUSIONS: Serum resistin is weakly associated with metabolic abnormalities in subjects with normal BMI, while in overweight/obese patients this correlation is not significant, perhaps due to the higher fat content in these subjects. Serum resistin is directly correlated with CRP and inversely to NT. An intriguing hypothesis, which needs to be tested, is that resistin is secreted in response to a chronic low-grade inflammation, and has antioxidant properties. PMID- 16044176 TI - Comparison of body fatness measurements by BMI and skinfolds vs dual energy X-ray absorptiometry and their relation to cardiovascular risk factors in adolescents. AB - OBJECTIVE: To compare estimates of adiposity by dual emission X-ray absorptiometry (DXA), skinfolds and body mass index (BMI); and to evaluate the relation of these measures to cardiovascular risk in adolescents. DESIGN: In a cohort of adolescents participating in a longitudinal study of insulin resistance, Slaughter formulas were used to estimate adiposity from skinfolds and DXA was used to estimate adiposity as % body fat (%BF) and fat mass (FBM). BMI, blood pressure, lipids and insulin resistance were measured. SUBJECTS: Male and female, 11-17 y old (n=130). MEASUREMENTS: To compare DXA with two office-based methods of assessing fatness and cardiovascular risk. RESULTS: Slaughter estimates were highly correlated with DXA (%BF r=0.92, P=0.0001; FBM r=0.96, P=0.0001). Correlations were similar in heavy and thin children. BMI was also highly correlated with DXA (%BF r=0.85, P=0.0001; FBM r=0.95, P=0.0001), and these relations were stronger in heavy than thin children. BMI and the Slaughter formulas were similar to DXA in their relations to cardiovascular risk factors. CONCLUSIONS: Adiposity by BMI and Slaughter formulas are highly correlated with DXA and similarly related to cardiovascular risk factors. BMI is easy to obtain and is an acceptable method for initial office estimation of body fatness. BMI and skinfolds compare well with DXA in predicting adverse cardiovascular risk profile. PMID- 16044177 TI - Pharmacogenomics in psychiatry: genomic considerations. PMID- 16044180 TI - The effect of Johne's vaccination on tuberculin testing in farmed red deer (Cervus elaphus). AB - AIM: To assess the degree of interference with bovine tuberculin testing in farmed red deer that vaccination of young deer with an oil-adjuvanted vs aqueous formulation of live attenuated Mycobacterium paratuberculosis Strain 316F vaccines would be likely to cause, and to compare immunological responses between vaccine formulations. METHODS: Five-month-old red deer (n = 45) were randomly allocated to three treatment groups of 15 animals, which received either no vaccine, a single 2-ml dose of an oil-adjuvanted formulation or two 2-ml doses, 6 weeks apart, of an aqueous formulation of live attenuated M. paratuberculosis Strain 316F vaccine injected subcutaneously (S/C) in the neck (Control, Oil adjuvant Ptb, and Aqueous Ptb groups, respectively). Injection- site reactions were described and measured on Weeks 3, 6 and 9. Animals were weighed and lymphocyte transformation tests (LTT) and antibody enzyme-linked immunosorbent assays (ELISA) using avian, bovine and Johnin tuberculin were conducted on blood samples collected at Weeks 0, 6, 12, 15, 24, 27, 36 and 39. A bovine mid-cervical skin test (MCT) was applied at Week 12, and comparative cervical skin tests (CCTs) at Weeks 24 and 36. At Week 42, the animals were slaughtered at a commercial deer slaughter premises and subjected to rigorous meat inspection. RESULTS: Two animals were eliminated at the start of the trial due to a positive cross-reaction with bovine tuberculin in the initial LTT. Almost all animals reacted to the MCT at Week 12, with mean skin thicknesses of 3.9, 2.9 and 1.0 mm for the Oil-adjuvant Ptb, Aqueous Ptb and Control groups, respectively. When the CCT was conducted at Week 24, 2/15 Oil-adjuvant Ptb, 2/14 Aqueous Ptb and 1/14 Control animals were classified as CCT-positive to bovine tuberculin. By Week 36, all animals were CCT-negative. The Oil-adjuvant Ptb vaccination resulted in high persistent levels of antibody that reacted with bovine tuberculin, compared with negligible levels in the Aqueous Ptb group. Overall, a single dose of the Oil adjuvant Ptb vaccine in deer stimulated a vigorous, cross-reactive immune response, evidenced by high LTT, skin-test and antibody reactions to bovine tuberculin, with both cell-mediated and humoral characteristics. By comparison, two doses of the Aqueous Ptb vaccine produced less cross-reactivity and a bias towards a cell-mediated response. The Oil-adjuvant Ptb vaccine resulted in moderate injection site lesions that were quite persistent, whereas the Aqueous Ptb vaccine resulted in smaller nodules that regressed more quickly. CONCLUSIONS: Vaccination of farmed deer with an oil-adjuvanted Johne's vaccine has the potential to cause significant interference with routine tuberculin skin testing. The cross-reactivity should decline with time and the CCT should be able to clear MCT-positives, but there is a risk of false-positives to the blood test for tuberculosis (BTB), due to high persistent levels of antibody. The CCT could be used as a primary skin test in vaccinated deer on some farms. The Aqueous Ptb caused fewer problems with skin testing and produced significantly less bovine antibody than the Oil-adjuvant Ptb, but stimulated persistent cell-mediated immune responses that may provide some protection against Johne's disease. PMID- 16044181 TI - A survey to investigate movements off sheep and cattle farms in New Zealand, with reference to the potential transmission of foot-and-mouth disease. AB - AIM: To quantify the numbers and extent of movements off sheep and cattle farms in New Zealand, in order to construct more realistic simulation models to investigate how infectious diseases such as foot-and-mouth disease (FMD) might spread. METHODS: Farmers from 500 randomly selected farms, comprising 100 from each of the following sectors, viz beef, dairy, grazing/dairy heifer rearing, sheep, and mixed sheep and beef, were asked to fill in diaries in which they recorded the movements of all animals, products, people, vehicles and equipment coming on to or leaving their farms during two separate 3-week periods, representing relatively 'busy' and 'quiet' times of the year with respect to livestock movements. Where possible, the destination of each movement was identified and geo-coded, to allow the distance travelled to be calculated. Each movement was then classified according to the risk of transfer of FMD virus (FMDV), should the disease have been present on the study farm at the time of the movement. The data were then analysed to establish movement frequencies and distributions of distances travelled, by the different pastoral livestock sectors. RESULTS: Two hundred and seventeen farmers returned one or more diaries. One hundred and ninety-three farmers completed a Busy-period diary, recording a total of 12,052 movements off their farms, a crude average of 62.4 per 3-week period, or 2.97 per day. Of these, 4.0% involved the transport of livestock, equating to 0.12 livestock consignments per day. In contrast, 186 Quiet-period diaries were returned, recording a total of 10,885 movements off, representing a crude average of 58.5 during the 3-week period, or 2.78 per day. Of these, 2.1% involved livestock, equating to 0.06 livestock consignments per day. The mean and median distances travelled during the Busy periods were 30.9 km and 13.1 km, respectively (range 0-1,167 km). In comparison, the mean and median distances travelled during Quiet periods were 41.3 and 14 km, respectively (range 0.4-1,203 km). CONCLUSIONS: People, vehicles, livestock and other items can travel off pastoral livestock farms in New Zealand to other farms either directly or via saleyards over extensive distances. This has implications for the potential spread of infectious diseases such as FMD. Movement parameters intended for use in the InterSpread Plus inter-farm simulation model of FMD were established, which will facilitate the prediction of likely spread and efficacy of controls in the unlikely event of a real-life outbreak. PMID- 16044182 TI - An in situ single-pass perfusion model for assessing absorption across the intestinal mucosa of the brushtail possum. AB - AIM: To develop an in situ animal model for assessing absorption of molecules across the intestinal mucosa of possums. METHODS: A surgical preparation was used to perfuse known concentrations of reference compounds (fluorescein and luteinising hormone-releasing hormone; LHRH) through measured sections of selected regions (jejunum, caecum, proximal colon) of the intestinal tract of 19 possums, over a 2-h period. Plasma concentrations of the compounds, which were perfused either with or without co-administration of a permeation enhancer (sodium deoxycholic acid; SDA), were determined in the perfusion effluent, peripheral and in some instances in the pre-hepatic circulation by spectrofluorometry (fluorescein) or radioimmunoassay (LHRH). Pharmacokinetic parameters of both compounds in the possum were determined over a period of up to 4 h in a further 30 animals (fluorescein, n = 6; LHRH n = 24), from their plasma profiles following intravenous (I/V) administration of a bolus dose. RESULTS: In animals perfused with 25 mg/ml fluorescein (Perfusion Experiment (PE) 1), the mean plasma concentration was 2.8 (SE 0.12) microg/ml in post-hepatic blood samples. When possums were perfused with 2.5 mg/ml fluorescein and 7 microg/ml LHRH (PE 2), mean plasma concentrations were 0.3 (SE 0.01) and 7.8 (SE 1.64) microg/ml fluorescein and 0.1 (SE 0.02) and 6.3 (SE 0.45) ng/ml LHRH, in the absence and presence of permeation enhancer, respectively. There was a poor correlation between pre-hepatic and post-hepatic concentrations. CONCLUSIONS: The single-pass in situ perfusion technique provided a useful model for investigating basic information on the absorption of biocontrol agents across the intestinal tract of possums, but had limitations that must be recognised. PMID- 16044183 TI - Cerebellar cortical abiotrophy in Wiltshire sheep. AB - AIM: To investigate the nature of a neurological disease in Wiltshire sheep. METHODS: Three affected lambs were examined, humanely killed and necropsied. Selected neurological tissues were examined by light and electron microscopy. RESULTS: Primary neurological lesions were confined to the cerebellum and were characterised by loss of Purkinje cells and the presence of large hypertrophied dendrites of surviving Purkinje cells. These contained stacks of smooth endoplasmic reticulum. There was hyperplasia and cell swelling of Bergmann glia. Mild Wallerian-type degeneration affected white matter in the cerebellum and spinal cord. CONCLUSION: The cerebellar lesions were of a degenerative and reactive rather than hypoplastic nature. These, and the history, suggest a genetic cause with putative inheritance as an autosomal recessive trait. Accordingly, the disorder is described as a cerebellar abiotrophy. PMID- 16044184 TI - Sucking behaviour of hand-reared newborn dairy calves. AB - AIM: To determine the feeding ability of calves during the first 4 days after birth. METHODS: The sucking behaviour of 171 dairy calves, fed from individual bottles during the first 4 days after birth, was evaluated by quantifying the volume of colostrum consumed, the duration of feeding, and speed of drinking. Calves had been separated from their mothers and brought into a rearing shed in the morning, when < 24 h of age, and were offered 2 L colostrum from a bottle in the afternoon of the same day, and twice daily thereafter. RESULTS: Newborn calves became efficient suckers from bottles within 24 h of removal from their dams (collection). On the day after birth, 95% of calves drank the 2 L of colostrum offered during the afternoon feed, and by Day 4 virtually all calves (99%) drank this amount. Calves that had inadequate colostrum from their dam were more likely to drink all 2 L offered after collection, but a small number of calves that had not had colostrum from their dams and drank < 500 ml at the afternoon feed following collection were likely not to drink 2 L on the following days. However, calves that did not drink all 2 L on Day 1 were not disadvantaged in comparison to those that did, in that they were equally likely to drink 2 L on Day 4. CONCLUSIONS: The majority of calves became efficient drinkers within 48 hours of birth, but a small number of slow feeders may need individual feeding at least up to 4 days after birth. PMID- 16044185 TI - The use of a medetomidine, butorphanol and atropine combination to enable blood sampling in young pigs. AB - AIM: To determine the suitability of a reversible, injectable anaesthetic combination including medetomidine, butorphanol and atropine to produce the degree of immobilisation required to allow blood sampling in young pigs. METHODS: Twenty 6-week-old crossbred, intact male pigs were sedated with an intramuscular (I/M) injection of 80 microg/kg medetomidine, 200 microg/kg butorphanol and 25 microg/kg atropine. Heart and respiratory rates and rectal temperatures were monitored. Excessive salivation, gagging, laryngeal reflex, presence of pedal reflex and deep and surface analgesia were noted. Time of injection and the time when pigs reached mild and full sedation were also recorded. RESULTS: Mild sedation was produced in 90% of pigs after 5.6 (SEM 0.96) min (n = 18; median 5, range 2-16 min), and full sedation (lateral recumbency and loss of jaw tone) in 60% of pigs after 12.5 (SEM 2.14) min (n = 12; median 10, range 5-28 min). The depth and duration of sedation were very variable and most animals were easily aroused. Ninety percent of the animals required the administration of halothane by mask to allow blood sampling, but the amount of halothane required was small. Heart and respiratory rates decreased (p < 0.001) but remained within the normal range. Rectal temperature was above normal at the time of sedation and at the time of blood sampling when the ambient temperature was 29 degrees C but not when the ambient temperature was reduced to 25 degrees C. CONCLUSIONS: The combination of medetomidine, butorphanol and atropine at these doses produced sedation of variable depth and duration that was insufficient on its own to allow blood sampling in the majority of pigs. Hyperthermia can occur in temperature controlled environments when using medetomidine, butorphanol and atropine in pigs. Reduction of stress and a quieter environment may improve the effects of the anaesthetic combination. PMID- 16044186 TI - Gross abnormalities, bacteriology and histological lesions of uteri of dairy cows failing to conceive or maintain pregnancy. AB - AIM: To describe the gross pathology, bacteriology and histopathology of the reproductive tracts of dairy cows that failed to conceive or maintain pregnancy. METHODS: The reproductive tracts of 105 cows that were not pregnant at the end of the seasonal breeding programme were retrieved following slaughter. The tracts were examined grossly, both horns of the uterine lumen were swabbed for bacteriology, and tissue was collected from each horn and the body of the uterus for histopathology. Grossly enlarged uteri and tracts containing purulent vaginal content were excluded. Histopathology was performed on three sections of uterine tissue from each of three cows in which no gross pathological changes were detected and from which no bacteria were isolated, from three cows in which no gross pathology was detected but from which bacteria were isolated, and from three cows in which gross pathological changes were detected but from which no bacteria were isolated. RESULTS: Thirty-six (34%) cows had one or more gross lesions which involved the ovary, uterine tube, uterus or vagina. Bacteria were isolated from the uteri of 22 (21%) cows. Isolates included Arcanobacterium pyogenes (n = 1), Escherichia coli (n = 1), Fusobacterium spp (n = 1), Haemophilus somnus (n = 5), Streptococcus acidominimus (n = 12), S. bovis (n = 2), S. uberis (n = 1) and S. salivarious (n = 1). In only five cows were both gross pathology and bacteria detected. There was no relationship between the isolation of bacteria and the diagnosis of gross pathology of the uterus. There were no differences in the degree of histopathological changes in the uteri from the three groups of cows examined, and lesions present were minor. CONCLUSIONS: Gross pathological changes and intra-uterine bacteria were found in 34% and 20% of cows, respectively, but the correlation between the two was poor. Histopathological changes were unremarkable, suggesting the bacteriological findings were coincidental, that causative agents of infertility were not present at the time of examination, or that unrelated causes such as nutritional anoestrus may have been responsible for the failure of some cows to conceive. PMID- 16044187 TI - Necrotising fasciitis associated with Escherichia coli in a dog. AB - CASE HISTORY: Lameness and limb pain associated with a laceration in the inner thigh of a Border Collie dog progressed over 4 days to extensive necrosis of the full-thickness of skin and subcutaneous (S/C) tissue. A successful outcome was achieved using surgical debridement and intensive supportive care, followed by limited local closure, axial pattern flap development, and free skin grafting. CLINICAL FINDINGS: Clinical findings included severe pain, depression, pyrexia and hypoalbuminaemia, and full-thickness loss of skin from the caudal thigh to the hock. Histopathologically, debrided tissue showed extensive necrosis of the dermis, adipose and muscle tissues, and the presence of numerous Gram-negative rods. Escherichia coli was cultured from deep tissue samples. DIAGNOSIS: Necrotising fasciitis (NF) associated with E. coli infection. CLINICAL RELEVANCE: NF associated with E. coli has not been previously reported in dogs. The importance of early diagnosis and surgical debridement is noted and the relevant medical literature is reviewed. PMID- 16044188 TI - An outbreak of Streptococcus canis mastitis in a dairy herd in Israel. AB - CASE HISTORY: An increase in the bulk somatic cell count (BSCC) of up to 1,000 x 103 cells/ml occurred in a dairy herd in Israel at the end of 2001 and beginning of 2002. CLINICAL FINDINGS: Bacteriological examination of milk from 69 cows revealed a high prevalence of Streptococcus group G bacteria, identified as S. canis, affecting 38% of cows and 20% of all quarters. Isolates were sensitive to cephalothin and moderately sensitive to penicillin G. Infected cows were separated from the herd, treated with intramammary antibiotics, milked last, and strict hygiene practices were introduced to the milking routine. The pathogen was cleared from the herd and BSCC decreased to 250-350 x 103 cells/ml after 6 months. DIAGNOSIS: Streptococcus canis mastitis. CLINICAL RELEVANCE: Streptococcus canis infection may cause subclinical mastitis and high bulk SCC in dairy herds and be resolved by treatment with intramammary antibiotics and the introduction of strict hygiene practices. PMID- 16044189 TI - Cobalt deficiency in sheep and diagnostic reference ranges. PMID- 16044190 TI - Crash emergency cesarean section: decision-to-delivery interval under 30 min and its effect on Apgar and umbilical artery pH. AB - PURPOSE: The objective of this study was to investigate the effect of decision-to delivery interval of crash emergency cesarean section on Apgar and umbilical artery pH in a level-3 university hospital. MATERIALS AND METHODS: In a retrospective analysis, all women undergoing "crash" emergency cesarean section were evaluated. Emergency cesarean sections were performed in the delivery room. Data relating to indication, Apgar score, arterial cord pH, and time intervals between decision-to-deliver and actual delivery were collected retrospectively. RESULTS: All 109 crash emergency cesarean sections were performed within a decision-to-delivery time of 30 min. The median (with 10-90th percentile) time was 10 min (5-19). Thirty-three (30.3%) of the emergency cesarean sections had a gestational age below 32 weeks and 60 (55%) below 37 weeks. An abnormal fetal heart rate pattern was noted in most of the cases (91%). Prolapsed cord (21%) and placental abruption (20%) were the most frequent reasons for emergency cesarean section but in one-fourth (25.7%) no morphological reason could be identified. Very short decision-to-delivery times below 20 min were inversely correlated to fetal outcome, i.e., lower umbilical blood pH and Apgar scores (P < 0.01). CONCLUSION: The 30-min standard for the decision-to-delivery time interval set by Anglo-American countries may be a feasible guideline at least for level-3 hospitals. The 20-min interval set by the German Society of Gynecology and Obstetrics could not be achieved in all cases. The positive effect of very short intervals on neonatal outcome still needs to be proven. PMID- 16044191 TI - Ureaplasma urealyticum-harmless commensal or underestimated enemy of human reproduction? A review. AB - This article reviews the role, diagnosis and treatment of Ureaplasma infections in human reproduction. PMID- 16044192 TI - A method for simple identification of signature peptides derived from polyUb-K48 and K63 by MALDI-TOF MS and chemically assisted MS/MS fragmentation. AB - A simple method is described to identify signature peptides derived from polyubiquitin (polyUb) chains. The method is based on MALDI-TOF MS/MS analysis after chemically assisted fragmentation, and works on peptides isolated from polyacrylamide gels. PolyUb chains branched at K48 and K63 were chosen as models for Ub-protein conjugates. They were resolved by SDS-PAGE, and their tryptic peptides (in-gel-trypsinolysis) derivatized with 3-sulfopropinic acid NHSester to obtain chemically assisted fragmentation during the MS/MS analysis. PolyUb-K63 produced a single peptide identified as (55)TLSDYNIQK(63) (GG)ESTLHLVLR(72). PolyUb-K48 produced two branched signature peptides identified as (43)LIFAGK(48)(GG)QLEDGR(54) and (43)LIFAGK(48)(LRGG)QLEDGR(54). The recovery of signature peptide with LRGG as branched chain underscores the need to take limited proteolysis into account in the search for detection of ubiquitinated peptides in proteomics studies. In conclusion, a simple method has been described allowing the identification of signature peptides, which are diagnostic markers of the majority of polyUb-conjugated proteins. In principle, the method should be applicable also for other more rare signature peptides. PMID- 16044193 TI - Using cellular automata images and pseudo amino acid composition to predict protein subcellular location. AB - The avalanche of newly found protein sequences in the post-genomic era has motivated and challenged us to develop an automated method that can rapidly and accurately predict the localization of an uncharacterized protein in cells because the knowledge thus obtained can greatly speed up the process in finding its biological functions. However, it is very difficult to establish such a desired predictor by acquiring the key statistical information buried in a pile of extremely complicated and highly variable sequences. In this paper, based on the concept of the pseudo amino acid composition (Chou, K. C. PROTEINS: Structure, Function, and Genetics, 2001, 43: 246-255), the approach of cellular automata image is introduced to cope with this problem. Many important features, which are originally hidden in the long amino acid sequences, can be clearly displayed through their cellular automata images. One of the remarkable merits by doing so is that many image recognition tools can be straightforwardly applied to the target aimed here. High success rates were observed through the self consistency, jackknife, and independent dataset tests, respectively. PMID- 16044194 TI - Synthesis of 1-(N-ethoxycarbonylamino)alkylphosphonic monoesters. AB - A series of 1-(N-ethoxycarbonylamino)alkylphosphonic monoesters were synthesized via three-component Mannich-type condensation of ethyl carbamate, aldehydes and dichlorophosphites in benzene, followed by hydrolysis. PMID- 16044195 TI - Analysis of amino acids in individual wheat embryonic protoplast. AB - Amino acids analysis in single wheat embryonic protoplast was performed using capillary electrophoresis equipped with laser-induced fluorescence (CE-LIF), combination with tissue culture technique. Reagent fluorescein isothiocyanate (FITC) was introduced into living protoplasts by electroporation for intracellular derivatization. A special osmotic buffer (0.6 mol/L mannitol, 5 mmol/L CaCl2) was used to keep the osmotic balance of embryonic protoplasts during the protoplasts derivatization. After completion of the derivatization reaction in the protoplasts, a single protoplast was drawn into the capillary tip by electroosmotic flow. Then a 0.1 M NaOH lysing solution was injected by diffusion. The derivatized amino acids were separated by capillary electrophoresis and detected by laser-induced fluorescence detection after the protoplast was lysed Nine amino acids were quantitatively and qualitatively determined and compared in lysate and single protoplast of wheat embryonic cells respectively, with mean concentrations of amino acids ranging from 2.68 x 10(-5) mol/L to 18.18 x 10(-5) mol/L in single protoplast. PMID- 16044197 TI - Tissue-water relations of two co-occurring evergreen Mediterranean species in response to seasonal and experimental drought conditions. AB - Tissue-water relations were used to characterize the responses of two Mediterranean co-occurring woody species (Quercus ilex L. and Phillyrea latifolia L.) to seasonal and experimental drought conditions. Soil water availability was reduced approximately 15% by partially excluding rain throughfall and lateral flow (water runoff). Seasonal and experimental drought elicited physiological and morphological adaptations other than osmotic adjustment: both species showed large increases in cell-wall elasticity and decreased saturated-to-dry-mass ratio. Increased elasticity (lower elastic modulus) resulted in concurrent decreases in relative water content at turgor loss. In addition, P. latifolia showed significant increases in apoplastic water fraction. Decreased saturated-to dry-mass ratio and increased apoplastic water fraction were accompanied by an increased range of turgor maintenance, which indicates that leaf sclerophyllous traits might be advantageous in drier scenarios. In contrast, the degree of sclerophylly (as assessed by the leaf mass-to-area ratio) was not related to tissue elasticity. An approximately 15% reduction in soil water availability resulted in significant reductions in diameter growth when compared to control plants in both species. Moreover, although P. latifolia underwent larger changes in tissue water-related traits than Q. ilex in response to decreasing water availability, growth was more sensitive to water stress in P. latifolia than in Q. ilex. Differences in diameter growth between species might be partially linked to the effects of cell-wall elasticity and turgor pressure on growth, since Q. ilex showed higher tissue elasticity and higher intrinsic tolerance to water deficit (as indicated by lower relative water content at turgor loss) than P. latifolia. PMID- 16044198 TI - Protein targeting into plastids: a key to understanding the symbiogenetic acquisitions of plastids. AB - Recent progress in molecular phylogenetics has proven that photosynthetic eukaryotes acquired plastids via primary and secondary endosymbiosis and has given us information about the origin of each plastid. How a photosynthetic endosymbiont became a plastid in each group is, however, poorly understood, especially for the organisms with secondary plastids. Investigating how a nuclear encoded plastid protein is targeted into a plastid in each photosynthetic group is one of the most important keys to understanding the evolutionary process of symbiogenetic plastid acquisition and its diversity. For organisms which originated through primary endosymbiosis, protein targeting into plastids has been well studied at the molecular level. For organisms which originated through secondary endosymbiosis, molecular-level studies have just started on the plastid targeted protein-precursor sequences and the targeting pathways of the precursors. However, little information is available about how the proteins get across the inner two or three envelope membranes in organisms with secondary plastids. A good in vitro protein-import system for isolated plastids and a cell transformation system must be established for each group of photosynthetic eukaryotes in order to understand the mechanisms, the evolutionary processes and the diversity of symbiogenetic plastid acquisitions in photosynthetic eukaryotes. PMID- 16044199 TI - R352Q mutation of the DHCR7 gene is common among Japanese Smith-Lemli-Opitz syndrome patients. AB - Smith-Lemli-Opitz syndrome (SLOS) is an autosomal recessive malformation syndrome characterized by microcephaly, syndactyly of toes, ambiguous genitalia, and mental retardation. The underlying DHCR7 gene has been identified and a wide variety of distinct mutations were reported in USA and European SLOS patients. A significant difference has been suggested in the frequency of SLOS among different ethnic populations. Here, we report mutational analysis of seven Japanese SLOS patients. Five mutations, R352Q, R242H, G303R, X476Q, and S192F, were identified, and R352Q appeared most frequent, since nine out of the 13 mutations of Japanese origin were the same R352Q. These results suggest that R352Q is a predominant founder mutation in Japanese SLOS patients. PMID- 16044200 TI - Strain-induced reorientation and mobility in nematic liquid-crystalline elastomers as studied by time-resolved FTIR spectroscopy. AB - Polarized Fourier transform infrared (FTIR) spectroscopy is employed to study the segmental orientation and mobility of liquid-crystalline elastomers (LCEs) with a monodomain structure in response to external mechanical fields parallel and perpendicular to the initial nematic director. The mean orientation and the molecular order parameter of the different molecular moieties referring to the mesogen, the spacer and the network are analyzed in detail. Parallel stretch leaves the mean orientation of the different molecular moieties and its molecular order parameter nearly uninfluenced. Perpendicular stretch results in a threshold like dependence: for elongation ratios lambda < or = lambda(c) = 1.3 (10 mol% crosslinker density), respectively lambda < or = lambda(c) = 1.6 (5 mol% crosslinker density) no change of the mean orientation and the molecular order parameters is observed, while for lambda > or = lambda(c) all molecular units reorient and their molecular order parameters are strongly decreased. The present studies give no indications that the reorientational dynamics of the network and the mesogens differ as long as the elongation ratio is smaller than lambda(c). PMID- 16044201 TI - Light-induced instabilities driven by competing helical patterns in long-pitch cholesterics. AB - We study theoretically the dynamical reorientation phenomena when a long-pitch cholesteric liquid-crystal film with homeotropic alignment is illuminated by a circularly polarized lightwave. In the present case, the natural cholesteric pitch is of the order of (or larger than) the film thickness. The helical cholesteric structure is thus frustrated by the boundary conditions without illumination. However, above a light intensity threshold reorientation occurs and the bifurcation scenario depends strongly on the natural cholesteric pitch. Recalling that a long-pitch cholesteric is achieved in practice by adding a small amount of chiral agents in a nematic liquid crystal, the observed dynamics can be viewed as the result of the competition between intrinsic and extrinsic unidimensional helical patterns. The intrinsic part consists of the helical deformations induced by the chirality of the dopant, whereas the extrinsic part is related to the chirality induced by the optical field through the non-uniform angular momentum transfer of light to a nematic. The all-optical analog in the case of a pure nematic (without chiral dopant), is also discussed. PMID- 16044202 TI - A prospective study between two different techniques for the repair of a large recurrent ventral hernia: a double mesh intraperitoneal repair versus onlay mesh repair. AB - BACKGROUND: Massive ventral hernias are difficult to repair, especially with multiple recurrences. Numerous methods of repair have been described with an overall recurrence rate up to 33% after first repair and 44% after second repair, mostly occurring within 3 years of the repair. METHODS: This is a prospective study on 41 patients who underwent surgery between January 2000 and August 2004 for recurrent large ventral hernias. Patients were randomized into two groups: group A included 22 patients, who were subjected to suture repair with an onlay polypropylene mesh, and group B, which included 19 patients, who were subjected to a tailored double mesh (Vicryl + polypropylene) intraperitoneal repair. RESULTS: Superficial wound infection occurred in two patients (4.8%), one in each group. By a median follow up of 30 months, seroma formation or hernia recurrence was not found in group B in comparison to seven and six cases, respectively, in group A (p < or = 0.000). There was no intraabdominal complication in the cases subjected to double mesh intraperitoneal repair due to the protective effect of the inner Vicryl layer, which is characterized by its low reactivity. CONCLUSIONS: A double mesh intraperitoneal repair (ADMIR) is successful for the repair of recurrent large ventral hernias as it is applicable to all sites of ventral hernias. The mesh is mostly hidden within the abdomen with relatively affordable pain allowing for early mobilization, the complication rate is low and so far no recurrence was reported. A long-term follow up with a larger number of cases is advisable in order to determine the long-term success of this kind of repair. PMID- 16044203 TI - Long-term follow-up after umbilical hernia repair: are there risk factors for recurrence after simple and mesh repair. AB - Adult umbilical hernia is a common surgical condition mainly encountered in the fifth and sixth decade of life. Despite the high frequency of the umbilical hernia repair procedure, disappointingly high recurrence rates, up to 54% for simple suture repair, are reported. Since both mesh and suture techniques are used in our clinic we set out to investigate the respective recurrence rates and associated complications, retrospectively. Patients who were treated between January 1998 and December 2002 were identified from our hospital database and invited to attend the outpatient department for an extra follow-up, history taking and physical examination. The use of prosthetic material, occurrence of surgical site infection, body mass and height as well as recurrence were recorded at the time of this survey. In total, 131 consecutive patients underwent operative repair of an umbilical hernia. Twenty-eight percent of the patients were female (n = 37). In 12 patients (11%) umbilical hernia repair was achieved with mesh implantation. Fourteen umbilical hernia recurrences were noted (13%); none had been repaired using mesh. No relationship was found between wound infection or obesity and umbilical hernia recurrence. In the light of these results it is necessary to re-evaluate our clinical "guidelines" on mesh placement in umbilical hernia repair: apparently not every umbilical fascial defect needs mesh repair. Research should focus on establishing risk factors for hernia recurrence. PMID- 16044204 TI - Vaginal pessaries in managing women with pelvic organ prolapse and urinary incontinence: patient characteristics and factors contributing to success. AB - OBJECTIVE: An aging population has resulted in higher prevalence of urinary incontinence (UI) and pelvic organ prolapse (POP). This study examines a nurse run clinic and analyzes the factors contributing to successful pessary use. STUDY DESIGN: A retrospective chart review of 1,216 patients was completed. History, pelvic examination and pessary fitting was done. Data was analyzed utilizing a categorical model of maximum-likelihood estimation to investigate relationships. RESULTS: Median patient age was 63 years. Median number of pessaries tried was two. Eighty-five percent of post-menopausal women were on hormone replacement therapy (HRT) prior to fitting. Highest success rate of 78% was in the group on both systemic and local HRT. Success rates ranged from 58% for urge incontinence to 83% for uterine prolapse. Prior vaginal surgery was a factor impacting success. In our series highest success rates for fitting were obtained with ring pessaries, ring with support, and gellhorns. CONCLUSIONS: This model is a viable, option for the conservative management of UI and POP. Local HRT plays an important role in successful pessary fitting. Complications are rare. PMID- 16044205 TI - Perineal ultrasound evaluation of the urethrovesical junction angle and urethral mobility in nulliparous women and women following vaginal delivery. AB - The objective of this study was to demonstrate the differences in the urethrovesical junction angle and urethral mobility by means of perineal ultrasounds in women following vaginal delivery with respect to nulliparous women. We have enrolled 34 women, 12 nulliparous (Group A) and 22 women in the post-partum period (Group B). The ultrasounds were employed to measure the urethrovesical junction angle ("urethro-pelvic" angle) and the urethral mobility. Data obtained show that the urethro-pelvic angle is narrower in Group A (97.3 degrees) with respect to Group B (112.5 degrees). Urethral mobility while coughing was significantly higher in Group B (30.4 degrees) with respect to Group A (-2 degrees). This study demonstrates the difference in the urethral support at rest and during cough in nulliparous versus recently delivered parous women. PMID- 16044206 TI - Aseptic meningitis and ischemic stroke in relapsing polychondritis. AB - Relapsing polychondritis (RP) is a rare disease characterized by recurrent inflammation of cartilaginous structures. Its involvement of central nervous system is uncommon. We report the case of a 71-year-old woman whose initial manifestations were RP and meningitis and whose subsequent attack of RP was associated with a left cerebral infarction. The patient responded to steroid therapy in both attacks of RP. PMID- 16044207 TI - A case of polyarteritis nodosa with lesions of the superior mesenteric artery illustrating the diagnostic usefulness of three-dimensional computed tomographic angiography. AB - A 40-year-old Japanese man who developed upper abdominal pain, weight loss, and hypertension was diagnosed as suffering with polyarteritis nodosa (PAN) with lesions of the superior mesenteric artery (SMA). Three-dimensional computed tomographic angiography (3D-CTA) and conventional angiography revealed smooth segmental luminal narrowing of the branches of the SMA. In addition, an enhanced abdominal CT scan demonstrated diffuse thickening of the wall of the affected SMA. Treatment with high-dose corticosteroids resulted in immediate improvement of the abdominal manifestations and normalized the serum C-reactive protein levels. Furthermore, the subsequent enhanced CT and 3D-CTA revealed improvement of the wall thickening and luminal narrowing of the SMA and its branches during the treatment period. In addition to possessing the diagnostic usefulness of conventional angiography, 3D-CTA is also less invasive and facilitates the prompt and accurate diagnosis of PAN. Furthermore, thickening of the wall of medium sized arteries evidenced by enhanced CT scan may also support a diagnosis of PAN. PMID- 16044208 TI - Ecological theory in practice: illustrations from a community-based intervention to promote the health of recent mothers. AB - We present a qualitative case study where we used four principles of ecological theory from community psychology as a template to assess the dynamics about how a preventive community intervention was transacted in eight communities in Victoria, Australia. The principles were cycling of resources, interdependence, adaptation, and succession. Ecological thinking focuses on key resources in communities. That is, the people, events, and settings that are the foundations of thinking about communities as systems. The data set consists of field diaries kept by and serial interviews with nine community development workers over a 2 year period. We found that the analysis highlighted a process-oriented way of representing the intervention, one that sees beyond the intervention's technical components (or packaged elements) to the complexities of the cultural and political change processes occurring beneath. The value of this is the attention focussed on likely project sustainability. PMID- 16044209 TI - Older adults' perspectives on screening prior to initiating an exercise program. AB - The purpose of this study was to explore the experiences of older adults with screening prior to starting an exercise program (structured or unstructured), and the impact this had on their willingness to engage in physical activity. This was a qualitative study using a focus group methodology. Focus groups were conducted in regionally dispersed sites in which exercise programs were ongoing or had been offered. A total of 122 older adults were included in this study, the majority of whom were African American (72%) with ages ranging from 50s to the 80s. An interview guide was used to facilitate each of the focus groups. Seventeen codes were identified and were reduced to 4 major themes: (1) Description of screening experienced; (2) Perception of purpose or need for screening; (3) Benefits of screening; and (4) Screening as a barrier to exercise. The participants indicated that there were some benefits associated with screening, such as safety assurances, although screening was also noted to be a barrier to exercising. Older adults who were already active, or had regular health care, believed that "screening" was not necessary. Pre-activity screening has different meanings for older adults depending on their own personal situations, resulting in potentially different motivational implications for adopting more active lives. For many, screening increased their sense of confidence and served as a positive motivator. Others felt that screening was irrelevant and actually hindered one's ability to join an activity group. We conclude with recommendations for refocusing future screening guidelines and suggestions for future research. PMID- 16044210 TI - Parent-teen disagreement of parent-imposed restrictions on teen driving after one month of licensure: is discordance related to risky teen driving? AB - The purpose of the investigation was to determine if parent-teen discordance for parent-imposed restrictions on driving conditions, driving rules, and the consequences for driving rule violations were related to risky teen driving. A total of 579 parents and their newly licensed teens were interviewed by telephone, 1 month after teens obtained provisional licenses. In multiple regression analyses, the degree of disagreement with parent restrictions on driving conditions and parent-imposed consequences for driving rule violations were negatively associated with a composite measure of teen risky driving. Female parents were negatively associated and male teens were positively associated with risky driving, but discordance with restricted driving conditions was the most important predictor. Discordance may reflect poor parent-teen relations or inadequate communication about parental expectations. The findings suggest that increasing parent-teen concordance on parent-imposed driving restrictions may help reduce risky teen driving. PMID- 16044211 TI - Differential effect of Huntington's and Parkinson's diseases in programming motor sequences of varied lengths. AB - BACKGROUND: Parkinson's disease (PD) and Huntington's disease (HD) patients have difficulties executing sequential movements. Attention control and short-term memory probably play an important role in programming sequential movements. To investigate the contribution of these cognitive factors to programming and executing visuomotor sequences in HD and PD patients a computerized version of the Corsi Block Tapping-Test was employed. METHODS: the performance of 11 patients with early stage PD, 11 HD patients with borderline to mild caudate atrophy and 20 healthy subjects was compared. The task was a reaction time task where targets were illuminated in groups of sequences increasing from 2 items to 5 items. Subjects reproduced the sequence (pressing the illuminated target) in the same order of appearance. Reaction Times and movement times were recorded. RESULTS: PD patients had increasing difficulties in programming and executing series greater than three components. HD patients did not differ significantly from the controls, although they showed a tendency to lose accuracy in the longer series. Both patient groups did not differ in their attention span. CONCLUSIONS: In PD although the spatial information may be well stored, they have difficulty accessing it when their attention is overloaded, leading to poor encoding and slow information processing. This process interferes with programming and execution of movement sequences. HD patients in the early stages of the illness seem to have more attention resources than PD patients, so that they start to show more problems in executing visuomotor sequences with longer movement sequences than PD patients. PMID- 16044212 TI - The window of therapeutic opportunity in multiple sclerosis: evidence from monoclonal antibody therapy. AB - From 1991-2002, we treated 58 patients with multiple sclerosis (MS) using the humanised monoclonal antibody, Campath-1H, which causes prolonged T lymphocyte depletion. Clinical and surrogate markers of inflammation were suppressed. In both the relapsing-remitting (RR) and secondary progressive (SP) stages of the illness, Campath-1H reduced the annual relapse rate (from 2.2 to 0.19 and from 0.7 to 0.001 respectively; both p < 0.001). Remarkably, MRI scans of patients with SP disease, treated with Campath-1H 7 years previously, showed no new lesion formation. However, despite these effects on inflammation, disability was differently affected depending on the phase of the disease. Patients with SPMS showed sustained accumulation of disability due to uncontrolled progression marked by unrelenting cerebral atrophy, attributable to ongoing axonal loss. The rate of cerebral atrophy was greatest in patients with established cerebral atrophy and highest inflammatory lesion burden before treatment (2.3 versus 0.7 ml/year; p = 0.04). In contrast, patients with RR disease showed an impressive reduction in disability at 6 months after Campath-1H (by a mean of 1.2 EDSS points) perhaps owing to a suppression of on-going inflammation in these patients with unusually active disease. In addition, there was a further significant, albeit smaller, mean improvement in disability up to 36 months after treatment. We speculate that this represents the beneficial effects of early rescue of neurons and axons from a toxic inflammatory environment, and that prevention of demyelination will prevent long-term axonal degeneration. These concepts are currently being tested in a controlled trial comparing Campath-1H and IFN-beta in the treatment of drug-naive patients with early, active RR MS. PMID- 16044213 TI - Impaired facilitation of motor evoked potentials in incomplete spinal cord injury. AB - OBJECTIVES: To improve the diagnosis of damaged spinal motor pathways in incomplete spinal cord injury (iSCI) by assessing the facilitation of lower limbs motor evoked potentials (MEP). METHODS: Control subjects (n = 12) and iSCI patients (n = 21) performed static and dynamic isometric foot dorsiflexions. MEPs induced by transcranial magnetic stimulation and EMG background of tibialis anterior muscle (TA) were analyzed. Static and dynamic muscle activation was performed at comparable levels of maximal voluntary contraction (MVC). The influence of the motor tasks on the excitability and facilitation of MEPs was compared between controls and iSCI patients. RESULTS: In the controls an increased facilitation of TA MEP at lower levels of dynamic compared with static activation (10-20% MVC) could be shown. At matched EMG background level the MEP responses were significantly increased. In the iSCI patients at a comparable level of TA activation the MEP responses were significantly reduced and 3 different patterns of MEP responses could be distinguished: i) preserved increment of TA MEP in the dynamic motor task, ii) unchanged MEP size in the dynamic and static motor task, and iii) elicitable MEPs in the dynamic motor task,which were abolished in the static motor task. CONCLUSIONS: Static and dynamic motor tasks have different effects on TA MEP facilitation. The task dependent modulation of TA MEPs is comparable to that described for upper limb muscles. Complementary to the MEP delay this approach allows for an estimation of the severity of spinal tract damage. The task-dependent modulation of TA MEPs is an additional diagnostic tool to improve the assessment and monitoring of motor function in iSCI. PMID- 16044214 TI - Chromosome 22q11.2 deletion syndrome: an underestimated cause of neuropsychiatric impairment in adolescence. PMID- 16044215 TI - The visual vertical in the pusher syndrome: influence of hemispace and body position. AB - The subjective visual vertical (SVV) was investigated in right brain-damaged (RBD) patients with pusher syndrome (PS) which is thought to stem from an erroneous perception of body orientation. The participants, sitting or lying, had to align a luminous rod with gravity. The task was performed in darkness with the rod centred to the body, or placed in the left (neglected) or in the right hemispace. The error, negligible in the control group (+0.3 degrees; n = 6) and mild in the nonneglect non-pusher patients (-1.8 degrees; n = 6), was clearly clockwise in the pusher neglect patients (N+P+; +7.2 degrees; n = 4), but anticlockwise in the non-pusher neglect patients (-6.6 degrees; n = 6). In both neglect groups, error was greater when the rod was in the left space. In N+P+ patients, the performance was strongly affected by posture (lying: +5.2 degrees ; sitting: +9.2 degrees ). Intra-individual variability was also much greater in this group. This study confirms the contralesional deviation of SVV in RBD patients without PS and suggests the presence of an opposite bias in RBD patients affected by PS. PMID- 16044216 TI - Positron emission tomography for radiation treatment planning. AB - PURPOSE: To evaluate the impact of positron emission tomography (PET) on target volume delineation for radiation treatment planning. MATERIAL AND METHODS: The data of the literature concerning the use of PET in target volume delineation are summarized. The following points are discussed for each tumor entity: biological background for the PET investigation, sensitivity and specificity of PET (with different tracers) in comparison to computed tomography (CT) and magnetic resonance imaging (MRI) and impact of PET on target volume definition. New PET tracers, which could visualize biological pathways, such as hypoxia, proliferation, angiogenesis, apoptosis and gene expression patterns, will also be discussed. RESULTS: The results of clinical studies on the integration of PET in target volume definition for lung, head-and-neck, genitourinary and brain tumors were analyzed. Fluorodeoxyglucose-(FDG-)PET has a significant impact on GTV (gross tumor volume) and PTV (planning target volume) delineation in lung cancer and can detect lymph node involvement and differentiate malignant tissue from atelectasis. In head-and-neck cancer, the value of FDG-PET for radiation treatment planning is still under investigation. For example, FDG-PET could be superior to CT and MRI in the detection of lymph node metastases and unknown primary cancer and in the differentiation of viable tumor tissue after treatment. Therefore, it might play an important role in GTV definition and sparing of normal tissue. Choline PET and acetate PET are promising tracers in the diagnosis of prostate cancer, but their validity in local tumor demarcation, lymph node diagnosis and detection of recurrence has to be defined in future clinical trials. FDG-PET seems to be particularly valuable in lymph node status definition in cervical cancer. In high-grade gliomas and meningiomas, methionine PET helps to define the GTV and differentiate tumor from normal tissue. For other entities like gastrointestinal cancer, lymphomas, sarcomas, etc., the data of the literature are yet insufficient. The imaging of hypoxia, cell proliferation, angiogenesis, apoptosis and gene expression leads to the identification of different areas of a biologically heterogeneous tumor mass that can individually be targeted using intensity modulated radiotherapy (IMRT). In addition, a biological dose distribution can be generated, the socalled dose painting. However, systematic experimental and clinical trials are necessary to validate this hypothesis. CONCLUSION: Regarding treatment planning in radiotherapy, PET offers advantages in terms of tumor delineation and the description of biological processes. To define the real impact of this investigation in radiation treatment planning, subsequent experimental, clinical and cost-benefit analyses are required. PMID- 16044217 TI - 12 years' experience with intraoperative radiotherapy (IORT) of malignant gliomas. AB - BACKGROUND: Even after surgery and radiotherapy, malignant gliomas still have a poor prognosis. The authors report on their experience with IORT in 71 patients. PATIENTS AND METHODS: From May 1992 to February 2004, 71 patients with malignant gliomas were treated with IORT. 26 patients suffered from grade III gliomas, 45 patients from glioblastomas (GBM). IORT was carried out using a standard electron tube and 9- to 18-MeV electrons. 52/71 patients who were primarily treated received 20 Gy IORT + 60 Gy postoperative radiotherapy, 19/71 patients with recurrences only received IORT (20-25 Gy). RESULTS: The complication rates were 1.4% for wound infections and 5.6% for hemorrhage. Median disease-specific survival amounted to 14.9 months (gliomas III) and 14.2 months (GBM). The 2-year survival rates amounted to 26.9% (gliomas III) and 6.8% (GBM; p = 0.0296). Total versus subtotal resection had no significant influence on survival (p = 0.0741), nor had age, sex, tumor site, performance status, size, primary versus recurrence, and radiation dose. A comparison to a conventionally treated patient group did not show a significant survival improvement. 3 months after treatment, initial symptoms had improved in 59% (hemiparesis), 50% (aphasia), 50% (hemianopsia), and 60% (convulsions). CONCLUSION: IORT has been shown to be feasible; perioperative complication rates were not increased. Survival was generally not improved compared to a historical control group. Recurrences achieved the same survival as primary tumors, and GBM also had a slightly increased survival, thus being possible indications for IORT. PMID- 16044218 TI - Optimizing cancer radiotherapy with 2-deoxy-d-glucose dose escalation studies in patients with glioblastoma multiforme. AB - BACKGROUND AND PURPOSE: Higher rates of glucose utilization and glycolysis generally correlate with poor prognosis in several types of malignant tumors. Own earlier studies on model systems demonstrated that the nonmetabolizable glucose analog 2-deoxy-D-glucose (2-DG) could enhance the efficacy of radiotherapy in a dose-dependent manner by selectively sensitizing cancer cells while protecting normal cells. Phase I/II clinical trials indicated that the combination of 2-DG, at an oral dose of 200 mg/kg body weight (BW), with large fractions of gamma radiation was well tolerated in cerebral glioma patients. Since higher 2-DG doses are expected to improve the therapeutic gain, present studies were undertaken to examine the tolerance and safety of escalating 2-DG dose during combined treatment (2-DG + radiotherapy) in glioblastoma multiforme patients. PATIENTS AND METHODS: Untreated patients with histologically proven glioblastoma multiforme (WHO criteria) were included in the study. Seven weekly fractions of (60)Co gamma rays (5 Gy/fraction) were delivered to the tumor volume (presurgical CT/MRI evaluation) plus 3 cm margin. Escalating 2-DG doses (200-250-300 mg/kg BW) were administered orally 30 min before irradiation after overnight fasting. Acute toxicity and tolerance were studied by monitoring the vital parameters and side effects. Late radiation damage and treatment responses were studied radiologically and clinically in surviving patients. RESULTS: Transient side effects similar to hypoglycemia were observed in most of the patients. Tolerance and patient compliance to the combined treatment were very good up to a 2-DG dose of 250 mg/kg BW. However, at the higher dose of 300 mg/kg BW, two out of six patients were very restless and could not complete treatment, though significant changes in the vital parameters were not observed even at this dose. No significant damage to the normal brain tissue was observed during follow-up in seven out of ten patients who received complete treatment and survived between 11 and 46 months after treatment. CONCLUSION: Oral administration of 2-DG combined with large fractions of radiation (5 Gy/fraction/week) is safe and could be tolerated in glioblastoma patients without any acute toxicity and late radiation damage to the normal brain. Further clinical studies to evaluate the efficacy of the combined treatment are warranted. PMID- 16044219 TI - [The influence of ionizing radiation on the development of posterior capsule opacification in vitro]. AB - BACKGROUND AND PURPOSE: Histologically, the posterior capsule opacification (PCO) corresponds to regenerative tissue of transformed lens epithelial cells (LECs) with extracellular matrix production. In this study, the influence of ionizing radiation on proliferating LECs and the development of PCO was investigated in vitro. MATERIAL AND METHODS: Each four and 14 pork lenses, respectively, were irradiated with 6 MeV electrons with single doses of 8, 10, 12, and 20 Gy. 1-2 h after irradiation the lens was removed by capsulorrhexis and hydrodissection. After fixation of the capsular bag in a special device the proliferation of residual LECs was examined daily. The experiment was considered to be finished when the capsular bag was completely opacified by confluent cell proliferates. RESULTS: Single dose irradiation with electrons in a dose range from 8 to 12 Gy significantly protracted the development of PCO with complete inhibition of PCO after application of 20 Gy. CONCLUSION: To inhibit PCO in vitro, a single dose of 20 Gy is necessary. The actual in vitro model allows an optimal investigation of PCO formation under different external influences and is therefore very suitable for radiobiological questions. PMID- 16044220 TI - [Quantitative and qualitative investigations of salivary gland function in dependence on irradiation dose and volume for reduction of xerostomia in patients with head-and-neck cancer]. AB - BACKGROUND AND PURPOSE: Radiation treatment of head-and-neck tumors mostly leads to a damage to the salivary glands and a consequential permanent loss of saliva. The aim of this investigation was to establish a modern three-dimensional conformal radiotherapy (3D-CRT) to show a decrease in severe xerostomia in contrast to the proven conventional technique (K-RT) with photons and electrons. PATIENTS AND METHODS: Between April 2002 and September 2003, 32 patients (25 male, seven female, mean age: 58 years) with malignant tumors of the head and neck were included-after surgery or in case of inoperability with curative intent in a prospective, nonrandomized study. 10/32 patients (31%) received K-RT with photons and electrons, and 22/32 patients (69%) 3D-CRT (six to eight photon portals). The quantity of saliva was measured as stimulated saliva flow rate (ml/5 min) prior to treatment, at the end, and 1, 6, and 12 months after termination of treatment. To find out the resulting mean dose of both parotid glands for every patient in Gray (D(mean) doses), the D(mean) doses of the ipsilateral and the contralateral parotid gland, determined by dose-volume histograms (DVHs), were averaged over. For calculation of the NTCP (normal tissue complication probability), the logistic model was used. RESULTS: In the trend the stimulated salivary flow rates were higher in the group with 3D-CRT than in the group with K-RT during the whole observation period (at 10 weeks after the start of radiotherapy 3D-CRT vs. K-RT with 1.56 +/- 1.6 vs. 0.82 +/- 1.2 ml/5 min; p < 0.1). The patients treated with the K-RT had, on average, significantly higher averaged D(mean) values than those irradiated with 3D-CRT (p < 0.012). Patients, who were irradiated with 3D-CRT for tumors of the larynx or hypopharynx, showed, on average, significantly lower D(mean) values than patients, who were treated with 3D-DRT because of oral cavity or oropharynx carcinomas or with K-RT irrespective of the primary tumor site (p < 0,003). The resulting dose for 50% complication probability (TD(50)) of the salivary glands was 36.9 Gy (30.9-43.5 Gy; 95% confidence interval). The gradient k of the curve located in point TD(50) was 7.7 (4.8-15.8; 95% confidence interval). CONCLUSION: Basically, 3D-CRT seems to be suitable as a standard for all patients with carcinomas of the oral cavity, oro- and hypopharynx. Especially in patients with tumors located in the larynx and hypopharynx, averaged D(mean) doses of both parotids during irradiation can be reached, to conserve salivary flow rates, which are similar to baseline flow rates. PMID- 16044221 TI - The influence of heterotopic ossification on functional status of hip joint following total hip arthroplasty. AB - PURPOSE: The functional failure induced by heterotopic ossification (HO) following total hip arthroplasty (THA) was analyzed and correlated to the radiologic failure. PATIENTS AND METHODS: From July 1997 to July 2001, 315 patients (345 hips) received THA indicated by a hypertrophic osteoarthritis of higher degree (Kellgren grade III, IV). All patients were irradiated prophylactically for prevention of HO on the evening before surgery with a 7-Gy single fraction. The patients' median age was 66.3 years. Radiologic failure was assessed by comparison of pre- and postoperative hip X-rays (immediately and 6 months after surgery). Analysis of radiographs was performed according to the Brooker Score. Clinical failure was appraised by measurement of passive range of motion (ROM) of the hip joint with a standard goniometer. The t-test was used for statistical analysis. RESULTS: 281 patients (81.5%) did not develop HO. HO of Brooker grade I or II was found in 58 patients (16.8%). Six patients (1.7%) developed HO Brooker grade III or IV. There was a significant negative correlation between the degree of radiologic and clinical failure. ROM differed significantly between patients with HO Brooker grade 0, I, II and patients with HO Brooker grade III, IV. Comparing the pre- and postoperative ROM, all patients with Brooker grade 0, I and II showed a significant improvement of flexion, internal and external rotation, abduction and adduction movement. Patients with HO Brooker grade III and IV showed no improvement of ROM in the postoperative follow-up. CONCLUSION: The development of HO following THA influences the physical function of the hip joint dependent on the degree of ossification. HO of lower degree (Brooker I, II) does not influence the clinical outcome, whereas HO of higher degree (Brooker III, IV) reduces the function of hip arthroplasty. Therefore, the purpose of a prophylactic therapy must be to reduce HO of higher degree. PMID- 16044222 TI - Ovarian function following pelvic irradiation in prepubertal and pubertal girls and young adult women. AB - PURPOSE: To analyze the effect of pelvic radiotherapy on ovarian function in prepubertal and pubertal girls and young adult women. PATIENTS AND METHODS: In a retrospective monoinstitutional analysis, patients < 30 years of age at diagnosis were included who had been irradiated between 1979 and 1998. The main tumor types were Hodgkin's disease (38%), Ewing's sarcoma (20%) and nephroblastoma (11%). Patients were classified into three groups according to the position of the ovary in relation to the radiation portals. Group 1 was defined by direct irradiation of both ovaries. Group 2 patients were included with both ovaries potentially located in the radiation portals. In group 3, at least one ovary was not directly irradiated. The median follow-up was 128 months. RESULTS: 16 of 55 analyzed patients were categorized in group 1. In ten of these patients, hormone status was evaluable. The ovarian doses were >/= 15 Gy. Except for one patient treated with 15 Gy all developed hormone failure. Eight of 14 patients of group 2 were evaluable. Seven of these patients developed ovarian failure. 19 of 24 patients in group 3 were evaluable. Nine of these patients developed ovarian failure. The observed difference in the rate of ovarian failure between the groups is statistically significant (p = 0.045). CONCLUSION: All patients receiving > 15 Gy to the ovaries developed hormone failure. In one case of a patient receiving an ovarian dose of 15 Gy, hormone failure was not found. In case of pelvic irradiation excluding at least one ovary, approximately half of the patients developed ovarian dysfunction, probably also due to the effects of polychemotherapy. PMID- 16044223 TI - Ultrafractionation does not improve the results of radiotherapy in radioresistant murine DDL1 lymphoma. AB - BACKGROUND AND PURPOSE: Low-dose hyperradiosensitivity (HRS), i.e., a relatively higher efficacy of doses < or = 0.5 Gy compared to doses > 1 Gy, has been shown in a number of tumor cell lines in vitro. Therefore ultrafractionated irradiation, i.e., application of very low doses per fraction, has been proposed to improve the effects of radiotherapy. The present study investigates ultrafractionation (UF) in radioresistant murine DDL1 T-cell lymphoma in mice. MATERIAL AND METHODS: UF was performed with 0.4 Gy per fraction, three fractions per day at 7 days per week, and conventional fractionation (CF) with 1.68 Gy per fraction, one fraction per day at 5 days per week. Tumor growth delay was evaluated for 2, 4 and 6 weeks of irradiation as time that tumors needed to reach fivefold the starting volume (GD(V5)). RESULTS: GD(V5) was not significantly different between UF and CF. The composite median relative GD(V5) calculated for all tumors irradiated in the present study was 1.00 [95% confidence interval 0.99; 1.08] in the CF and 0.99 [0.92; 1.01] in the UF arm (p = 0.24). CONCLUSION: UF was not more efficient than CF in DDL1 tumors. Taken together with previous experiments on human A7 glioblastoma, which showed a negative effect of UF on local tumor control, the preclinical data obtained in this laboratory so far do not support the use of ultrafractionated schedules in radiotherapy. PMID- 16044224 TI - Radiation therapy and simultaneous chemotherapy for recurrent cervical carcinoma. AB - PURPOSE: To evaluate the efficacy and toxicity in patients with recurrence of cervical cancer treated with radiotherapy and simultaneous chemotherapy. PATIENTS AND METHODS: Between 1987 and 2001, 24 patients with recurrent cervical carcinoma were treated with concurrent chemoradiotherapy. Nine patients had incomplete tumor resection prior to radiation therapy. Irradiation was delivered to a total dose of 60 Gy, in three patients with central recurrences supplemented by brachytherapy. One patient was treated with brachytherapy alone. Simultaneous chemotherapy was done as a combined therapy of 5-fluorouracil-(5-FU, 600 mg/m(2)/d1-5, 29-33) and cisplatin (20 mg/m(2)/d1-5, 29-33; 16/24 patients) or of 5-FU (1,000 mg/m(2)/d1-5, 29-33) and mitomycin C (10 mg/m(2)/d2, 30; 1/24 patients). Cisplatin alone (25 mg/m(2)/d1-5) and carboplatin alone (800 mg/m(2)/d1-5) were administered in 5/24 patients (21%) and 2/24 patients (8%). RESULTS: The 5-year local recurrence-free survival rate was 37%, disease-free survival 33%, and overall survival 34%. Grade 3 toxicity (NCI-CTC grade 3) occurred mainly as diarrhea (38%), leukopenia (33%), and nausea (21%). Severe toxicity (grade 4) was not seen in any of the patients. CONCLUSION: Radiation therapy with simultaneous chemotherapy for recurrences of cervical cancer is an effective treatment with acceptable toxicity. PMID- 16044225 TI - Evaluation of speech disorders in children with cleft lip and palate. AB - BACKGROUND: We investigated the reliability of nasopharyngoscopy and auditory perception, two common clinical methods employed in examining the speech pathology of children with cleft lip and palate. PATIENTS AND METHODS: Nasopharyngoscopy was performed to assess velopharyngeal closure function and nasopharyngeal morphology. The auditory examination evaluated nasality and other speech disorders such as articulatory tension, shift in articulation, and sigmatism. The study was based on the audio and video recordings of twelve patients (9.5 +/- 0.5 years) after surgical rehabilitation of their cleft lip and palate. The speech sample consisted of reading a standardized sequence of words. The recordings were analyzed by several examiners with different experience in the evaluation procedure at two time-points. Inter-rater and intra-rater reliability was determined by Cohen's Kappa coefficient and by estimation of the 95% intervals of confidence. RESULTS: Analysis of the visual and auditory perceptual findings by the experienced raters was highly reliable. However, the less experienced raters' results showed greater variability, as did morphological evaluation in both groups. Comparison between visual and auditory evaluations concerning hypernasality resulted in little correlation. CONCLUSIONS: These two subjective methods of evaluation are recommended because of their high reliability with experienced raters. Our results suggest that "experience" is an essential criterion in determining the structural quality of the morphological and functional assessment of typical speech pathologies in children with cleft lip and palate. In addition, the data demonstrate the necessity of special multidisciplinary rehabilitation teams in so-called cleft centers. PMID- 16044226 TI - Magnetic field interactions of orthodontic wires during magnetic resonance imaging (MRI) at 1.5 Tesla. AB - BACKGROUND: Orthodontic appliances pose a potential risk during magnetic resonance imaging (MRI) due to forces on metallic objects within the static magnetic field of MRI systems. The aim of the present investigation was to measure forces on orthodontic wires caused by the static magnetic field of a 1.5 Tesla MRI system, and to assess the safety hazards associated with these forces. MATERIALS AND METHODS: Thirty-two different orthodontic wires (21 arch wires, eight ligature wires and three retainer wires) were investigated in a 1.5-Tesla MRI system (Magnetom Symphony, Siemens Medical Solutions, Erlangen, Germany). The translational forces were measured using the deflection angle test (ASTM F2052 02); rotational forces were assessed on a 5-point qualitative scale. RESULTS AND CONCLUSION: All retainer wires and the steel arch wires (the Noninium arch wire being the exception) were subjected to considerable rotational and translational forces within the MRI system's magnetic field. Translational forces were from 9.1 to 27.6-times as high as gravitational forces on these objects. Steel ligature wires and arch wires made of cobalt chromium, titanium molybdenum, nickel titanium, and brass alloys showed no or negligible forces within the magnetic field. The translational and rotational forces within the MRI magnetic field should pose no risk to carefully-ligated arch wires. Steel retainer wire bonds should be checked to ensure secure attachment prior to an MRI investigation. PMID- 16044227 TI - Investigation of maxillary tooth sizes in patients with palatal canine displacement. AB - AIM: The aim of this retrospective trial was to investigate differences in mesiodistal and vestibulo-oral crown sizes of naturally, fully-erupted permanent maxillary teeth between patients with and without palatal canine displacement. PATIENTS AND METHOD: 115 patients (mean age: 14 years 10 months; females: 77 males: 38) treated in the Department of Orthodontics, University of Munich were included in the study. 65 of the patients showed at least one palatally-displaced canine. Diagnosis and the location of the displacement were determined on the basis of standardized radiographs and confirmed by surgical documentation. Each maxillary tooth's mesiodistal and vestibulo-oral width was measured using a dial caliper on each dental cast. Excluded were partially-erupted teeth and surfaces with caries or restorations that had to be measured. An analysis of available space was made by evaluating the pre-treatment dental casts of all patients included in the study. RESULTS: Comparing the tooth widths of patients with unilateral canine displacement with the corresponding contralateral quadrants, we noted a statistically significant difference, namely that the central and lateral incisors and the canines of the affected side were narrower than those of the non affected side in the same patient. Moreover, the displaced upper canines showed an increase in vestibulo-oral dimension. Overall tooth width (including all tooth groups) in patients with palatally-displaced canines was significantly less than that in the control group. However, when comparing the crown diameters of unilaterally- and bilaterally-affected patients, no differences in tooth-size were observed. The space-analysis showed excessive dental-arch space in patients with a palatally-displaced canine. CONCLUSION: Patients affected by palatal canine displacement showed significantly smaller maxillary tooth size. PMID- 16044228 TI - Dental care with manual toothbrushes during fixed orthodontic treatment--a new testing procedure. AB - AIM: The aim of this investigation was to employ a new in-vitro testing system for manual toothbrushes in order to distinguish the more effective from those less so for dental care during fixed appliance treatment. MATERIALS AND METHODS: The testing apparatus consisted of a sliding carriage able to execute a horizontal brushing movement, and a row of artificial teeth upon which the various toothbrushes were manipulated. The artificial row of teeth was fixed on a sensor that recorded in all three dimensions the forces and moments caused by the toothbrushes on the toothbrush field. All the tests were executed with a weight of 110 g on a tooth field with a multibracket appliance. Tests were also carried out with five toothbrushes having weights of 200 g, 250 g and 300 g. Here, the decisive target values were 1) the degree of exertion necessary in the brushing direction to move a brush over the artificial teeth, and 2) the maximum force occurring in the brushing direction. High target values indicated high interaction between toothbrush bristles and the surfaces being brushed. RESULTS: From testing five toothbrushes with four different weights, we have established profiles confirming the beneficial and less beneficial properties of certain toothbrushes involving various high contact forces. PMID- 16044229 TI - Standardization of orthodontic products--does it make sense? AB - Since 1994, the DIN (Deutsches Institut fur Normung e.V.) has had a German working group named "Orthodontic Products" working toward establishing standardized norms for orthodontic products. In the USA, a similar working group called "Orthodontic Materials" was established in 1996 at the ADA (American Dental Association). Working at the ISO level since 1997, an international team has been in charge of unifying the standards already set at national levels. It is the aim of this article to report on the current state of standardization in Germany and internationally. The standards already published for wires (DIN 13971, ADA Spec. 32), for brackets and tubes (DIN 13971-2, ADA Spec. 100) and for elastomeric elements (DIN 13901) have been considered, as have problems occurring with ISO standards (ISO/CD 15841 and ISO/CD 15841-2) and DIN 13904-1. We demonstrate that the demands made of these products by the orthodontists who use them have lead to reasonable quality standards. Thanks to clearly-defined testing methods, orthodontic products can now be tested and their compliance with these minimum standards can be compared. In fact, the standards that have been agreed upon help both the product manufacturers and their users to improve product quality and, ultimately, the success of orthodontic treatment. PMID- 16044230 TI - Management of an extremely displaced maxillary canine. AB - CASE REPORT: Aligning a displaced maxillary canine into the dental arch is one of the most complicated problems in orthodontics. In cases of extremely high displacement, the tooth is frequently removed surgically. Because of the upper canines' significance to dental esthetics and functional occlusion, such a decision is a very serious one. This case report illustrates the treatment of an extremely high displaced maxillary canine. CLINICAL FINDINGS: The main diagnosis was the displacement and the retention of tooth 13 (in nearly horizontal position, apical to the neighboring teeth); further diagnoses were: transversal maxillary deficiency with frontal crowding and a distal bite of one premolar in width, a deep bite of 6 mm with contact in the palatal mucosa, mandibular midline deviation of 2.5 mm to the right, lingual eruption of teeth 32 and 42, retroinclination of the maxillary incisors, and retarded eruption of the permanent teeth. THERAPY: Initial treatment with active and functional appliances to correct the distal bite, midline deviation and deep bite. Surgical exposure of the high displaced canine at the age of 14. Onset of cuspid elongation with removable appliances and elastics, further movement with a transpalatinal bar and welded arm, and full alignment of the upper and lower arches with fixed appliances in both jaws. Stabilization of the orthodontic treatment results with retention devices. Duration of treatment: 5 years and 8 months. For the alignment of tooth 13, 2 years and 10 months were required; 1 year and 4 months were necessary with complete fixed appliance. CONCLUSIONS: The aim of this case report was to demonstrate the potential of aligning an extremely displaced canine. Because of the esthetic and functional importance of the upper canines, therapeutic alignment should be initiated, provided there are no indications to the contrary. PMID- 16044231 TI - [Myocardial preconditioning with volatile anesthetics. General anesthesia as protective intervention?]. AB - Reduction of the perioperative cardiovascular risk with pharmacological interventions plays a prominent role in routine anesthesia practice. For example, perioperative beta-blockade is well established in anesthesiological treatment of patients. There is a growing body of evidence supporting the cardioprotective effects of volatile anesthetics known as anesthetic-induced preconditioning. There are numerous and complex data from animal studies. The mechanisms of anesthetic-induced preconditioning have been extensively studied but have still not been clearly identified. Initial clinical data show the cardioprotective effects of volatile agents by looking at parameters of myocardial function and laboratory values and therefore, the question of the relevance of these data for routine clinical practice has been raised. This review gives a summary of the currently available data focusing on the mechanisms of anesthesiological preconditioning and clinical studies. PMID- 16044232 TI - [Analgesia with remifentanil in spontaneously breathing patients undergoing brief but painful radiological procedures]. AB - INTRODUCTION: Percutaneous transhepatic biliary drainage (PTBD) and stenting are very painful procedures in interventional radiology and require potent analgesia; employing remifentanil in spontaneously breathing patients may be one possible strategy. PATIENTS AND METHODS: The study group was composed of 18 men and 2 women with a mean age of 63+/-10 (mean+/-SD) years. Pain intensity was measured with a VAS score before the procedure, after local anesthesia on the rib cage, after stenting and after the radiology procedure. RESULTS: Remifentanil infusion (dosage: 0.12-0.30 microg/kg body weight/min) was infused throughout the entire radiology procedure according to physical status, past medical history, individual pain, and clinical assessment. During insufflation of 10l O(2)/min via a venturi mask, oxygen saturation did not fall below 96% at any time-point during the procedure. In the VAS score, we noted a decrease after starting the remifentanil infusion towards the end of procedure. All patients were able to move into bed without help. Postoperatively, no analgesics and no antiemetics were needed. CONCLUSIONS: Employing a remifentanil infusion for brief interventional radiology procedures in palliative treatment of patients resulted in high patient and radiologist comfort. PMID- 16044234 TI - Extensive traction alopecia attributable to ponytail hairstyle and its treatment with hair transplantation. PMID- 16044235 TI - Secondary rhinoplasty. AB - Revision rhinoplasty is a complex aesthetic and reconstructive procedure in which both functional and cosmetic principles must be considered in the planning of an appropriate operation. Different techniques must be modified according to the specific defects. The modifications may vary from simple integration of a previous poorly performed surgery to complex grafting of homologous or heterologous material. The authors report their experience with 311 cases of revision rhinoplasty. PMID- 16044236 TI - Isolated congenital absence of the nasal bones and aesthetic surgical correction: managing and case report. AB - A rare case of isolated congenital bone agenesis in a young woman referred to the authors for aesthetic correction of the dorsum profile is described. There are no descriptions of surgical treatment for this isolated facial defect in the literature. After a complete and accurate diagnostic study, a rhinoplasty was performed with a good result and without complications during a 2-year follow-up period. To explain this congenital malformation, several hypotheses were examined, such as bony resorption and embryologic bony development syndromes. This report describes the possibility of intervention in the case of nasal bone absence and shows how risks of structural instability can be avoided with an accurate preoperative study and a careful surgical approach. PMID- 16044237 TI - Filler injection enhances the projection of the reconstructed nipple: an original easy technique. AB - BACKGROUND: Reduced nipple projection is the main reason for unsatisfactory nipple-areola complex reconstruction, and many techniques have been proposed to maintain projection of the reconstructed nipple. METHODS: For 70 patients, 90 nipples were reconstructed using either a small wedge from the labia minora (LMW) (n=70) or nipple sharing (NS) (n=20). Two months after reconstruction, each reconstructed nipple was injected with DermaLive. Second and third injections were performed 2 and 5 months later. The injected volume was tailored to the desired projection. Nipple projection was measured at the moment of implant, before and after each injection, and 6 and 12 months after the last injection. RESULTS: Nipple projection was satisfactory in all cases and comparable with that of the contralateral nipple. The average nipple projection at 6 months was 5.8 mm in the LMW group and 3.8 mm in the NS group (p<0.01) and, respectively, 5.6 mm and 3.5 mm at 12 months (p<0.01). No complications occurred, except for one positron emission tomography (PET) false-positive result. CONCLUSIONS: The described method is simple and safe. It provides precise projection with no need for intraoperative forecasting of tissue reabsorption. The result was better for the LMW patients, perhaps because of their higher distensibility. PMID- 16044238 TI - A perichondrial flap for functional purposes in rhinoplasty. AB - Recent trends in rhinoplasty techniques have focused on anatomic repair as well as respect and preservation of soft tissue integrity. In this article, the authors describe the use of a perichondrial flap, then discuss technical considerations and clinical perspectives of their advantages. A perichondrial flap helps to restore the stability of the upper lateral cartilage, to achieve extra padding, and to secure osteocartilagenous grafts. The method described has been used for 60 consecutive patients. The majority of these patients were satisfied with the results. To obtain detailed information about the perichondrial flap, the authors performed an anatomic study of 13 cadavers. The average thickness of the perichondrium was 186+/-146.1 microm (range, 90--596 microm). On the basis of the results, it was concluded that elevation of the perichondrial flap with loupe magnification could improve the outcome of rhinoplasty. PMID- 16044239 TI - Question mark ear deformity and a modified surgical correction method: a case report. AB - Question mark ear (Cosman ear) deformity, a rare congenital malformation, is characterized by a cleft between the helix and the ear lobe and marked prominence of the auricles. Although the features of question mark ear deformity are well described in the literature, there is no definitive surgical technique for repair of this deformity, Several surgical methods have been introduced for the correction of the deformity. These techniques mainly provide for repair of the cleft between the helix and ear lobe. However, marked prominence of the upper auricle usually is also present with the cleft. We modified the surgical technique to correct the cleft and the upper prominence at the same time. With this procedure, the cleft is exposed by raising a vertical cutaneous flap based on the cleft on the posterior side of the ear, After anterior scoring to form the antihelix and cleft repair using an ipsilateral conchal cartilage graft, the cutaneous flap is used to cover the cartilage graft and the flap donor site is closed primarily to facilitate restoration of the antihelix. The authors report on a patient with Cosman ear and introduce their modified technique that can be used for repair of the cleft between the helix and ear lobe and the prominence of the upper helix in the same procedure. PMID- 16044240 TI - Myopic change after transconjunctival blepharoplasty using carbon dioxide laser: case report. AB - The case of a patient who experienced temporary myopic change after transconjunctival blepharoplasty using the carbon dioxide (CO(2)) laser is reported. Transconjunctival lower lid blepharoplasty using the CO(2) laser, one of the most frequent laser procedures performed in aesthetic surgery, is considered to be a safe and reliable approach that does not incur significant complications. This report describes temporary myopia in a 38-year-old woman as a rare complication after this procedure. As for the possible etiology of the postoperative myopia, the authors strongly suspect a temporary mild deformity of the eyeball leading to the change in refraction, including scleral edematization and/or retinal indentation attributable to thermal injury as a result of CO(2) laser irradiation adjacent to the eyeball. The purpose of this case report is to inform others of the complication the authors encountered and to discuss the possible etiology. PMID- 16044241 TI - Proteome composition in Plasmodium falciparum: higher usage of GC-rich nonsynonymous codons in highly expressed genes. AB - The parasite Plasmodium falciparum, responsible for the most deadly form of human malaria, is one of the extremely AT-rich genomes sequenced so far and known to possess many atypical characteristics. Using multivariate statistical approaches, the present study analyzes the amino acid usage pattern in 5038 annotated protein coding sequences in P. falciparum clone 3D7. The amino acid composition of individual proteins, though dominated by the directional mutational pressure, exhibits wide variation across the proteome. The Asn content, expression level, mean molecular weight, hydropathy, and aromaticity are found to be the major sources of variation in amino acid usage. At all stages of development, frequencies of residues encoded by GC-rich codons such as Gly, Ala, Arg, and Pro increase significantly in the products of the highly expressed genes. Investigation of nucleotide substitution patterns in P. falciparum and other Plasmodium species reveals that the nonsynonymous sites of highly expressed genes are more conserved than those of the lowly expressed ones, though for synonymous sites, the reverse is true. The highly expressed genes are, therefore, expected to be closer to their putative ancestral state in amino acid composition, and a plausible reason for their sequences being GC-rich at nonsynonymous codon positions could be that their ancestral state was less AT-biased. Negative correlation of the expression level of proteins with respective molecular weights supports the notion that P. falciparum, in spite of its intracellular parasitic lifestyle, follows the principle of cost minimization. PMID- 16044242 TI - PAQR proteins: a novel membrane receptor family defined by an ancient 7 transmembrane pass motif. AB - An emerging series of papers has identified new receptor proteins that predict seven-transmembrane pass topologies. We have consolidated this family to 11 human genes and have named the family PAQR, after two of the initially described ligands (progestin and adipoQ receptors). This protein family has ancient evolutionary roots, with identified homologs found in eubacteria. To date, published data indicate that the prokaryotic members of this family appear to encode hemolysin-type proteins, while in eukaryotes, PAQR proteins encode functional receptors with a broad range of apparent ligand specificities. We provide the complete human and mouse complement of this family, suggest a conserved structure/topology with invariant intracellular amino acid residues, and have measured mRNA expression levels for these genes across a range of human tissues. PMID- 16044243 TI - Actin phylogeny and intron distribution in bangiophyte red algae(rhodoplantae). AB - The molecular phylogeny of red algal actin genes, with emphasis on the paraphyletic "Bangiophyceae," was examined and compared to the rhodophyte SSU rDNA phylogeny. Nineteen new genomic actin sequences and seven SSU rDNA sequences were obtained and subjected to diverse phylogenetic analyses (maximum likelihood, distance/neighbor-joining, maximum parsimony, Bayesian analyses, and, with respect to protein sequences, also quartet puzzling). The actin trees confirmed most of the major clades found in the SSU rDNA phylogenies, although with a lower resolution. An actin gene duplication in the florideophycean lineage is reported, presumably related to an increased complexity of sexual reproduction. In addition, the distribution and characteristics of spliceosomal introns found in some of the actin sequences were examined. Introns were found in almost all florideophycean actin genes, whereas only two bangiophyte sequences contained introns. One intron in the florideophycean actin genes was also found in metazoan, and, shifted by one or two nucleotides, in a glaucocystophyte, a cryptophyte, and two fungal actin genes, and thus may be an ancient intron. PMID- 16044244 TI - The RNA world on ice: a new scenario for the emergence of RNA information. AB - The RNA world hypothesis refers to a hypothetical era prior to coded peptide synthesis, where RNA was the major structural, genetic, and catalytic agent. Though it is a widely accepted scenario, a number of vexing difficulties remain. In this review we focus on a missing link of the RNA world hypothesis-primitive miniribozymes, in particular ligases, and discuss the role of these molecules in the evolution of RNA size and complexity. We argue that prebiotic conditions associated with freezing, rather than "warm and wet" conditions, could have been of key importance in the early RNA world. PMID- 16044245 TI - Biases in phylogenetic estimation can be caused by random sequence segments. AB - We consider the effects of fully or partially random sequences on the estimation of four-taxon phylogenies. Fully or partially random sequences occur when whole subsets of sequences or some sites for subsets of sequences are independent of sequence data for the other taxa. Random sequences can be a consequence of misalignment or because sites evolve at very fast rates in some portions of a tree, a situation that occurs especially in analyses involving deep divergence times. One might reasonably speculate that random sites will only add noise to the estimation of a phylogeny. We show that in the case that a random sequence is added to a three-taxa alignment, it is more likely to be a neighbor of the sequence corresponding to the longest branch in the three-taxon tree. Surprisingly, when only about half of the sites show randomness, a long-branch repels form of small sample bias occurs, and when a minority of sites show randomness this becomes a long-branch-attraction bias again. The most serious bias, one that does not vanish with increasing sequence length, occurs when more than one sequence is partially random. If there is a large amount of overlap in the random sites for two sequences, those two sequences will be attracted to each other; otherwise, they will repel each other. Random sequences or sites can, therefore, cause complicated biases in phylogenetic inference. We suggest performing analyses with and without potentially saturated sequences and/or misaligned sites, to check that these biases are not affecting the inferred branching pattern. PMID- 16044246 TI - Analysis of recombination in Campylobacter jejuni from MLST population data. AB - We analyze recombination in C. jejuni using MLST data from isolates taken from wild birds, cattle, wild rabbits, and water in a 100-km2 study region in Cheshire, UK. We use a recent approximate likelihood method for inference, based on combining likelihood information from all pairs of segregating (polymorphic) sites in the data. We find substantial evidence for recombination, but only for recombination with short tract lengths, of around 225-750 bp. We estimate that the rate of recombination is of a similar magnitude to the rate of mutation. PMID- 16044247 TI - Positively selected sites in the Arabidopsis receptor-like kinase gene family. AB - We analyze members of the receptor-like kinase (RLK) gene family in Arabidopsis thaliana for positive selection. Likelihood analyses find evidence for positive selection in 12 of the 52 RLK family sequences groups. These 12 groups represent 97 of the 403 sequences analyzed. The majority of genes in groups subject to positive selection have not been functionally characterized, but sites under selection are predominantly located in the extracellular region. The pattern of selection in the extracellular leucine-rich repeat (LRR) motif of groups 14 and 51 is similar to previous studies where positively selected positions are located in a solvent exposed beta-strand that may determine disease specificity, raising the possibility that some RLK genes function in a similar role. PMID- 16044248 TI - Codon bias and noncoding GC content correlate negatively with recombination rate on the Drosophila X chromosome. AB - The patterns and processes of molecular evolution may differ between the X chromosome and the autosomes in Drosophila melanogaster. This may in part be due to differences in the effective population size between the two chromosome sets and in part to the hemizygosity of the X chromosome in Drosophila males. These and other factors may lead to differences both in the gene complements of the X and the autosomes and in the properties of the genes residing on those chromosomes. Here we show that codon bias and recombination rate are correlated strongly and negatively on the X chromosome, and that this correlation cannot be explained by indirect relationships with other known determinants of codon bias. This is in dramatic contrast to the weak positive correlation found on the autosomes. We explored possible explanations for these patterns, which required a comprehensive analysis of the relationships among multiple genetic properties such as protein length and expression level. This analysis highlights conserved features of coding sequence evolution on the X and the autosomes and illuminates interesting differences between these two chromosome sets. PMID- 16044249 TI - Global mRNA stability is not associated with levels of gene expression in Drosophila melanogaster but shows a negative correlation with codon bias. AB - A multitude of factors contribute to the regulation of gene expression in living cells. The relationship between codon usage bias and gene expression has been extensively studied, and it has been shown that codon bias may have adaptive significance in many unicellular and multicellular organisms. Given the central role of mRNA in post-transcriptional regulation, we hypothesize that mRNA stability is another important factor associated either with positive or negative regulation of gene expression. We have conducted genome-wide studies of the association between gene expression (measured as transcript abundance in public EST databases), mRNA stability, codon bias, GC content, and gene length in Drosophila melanogaster. To remove potential bias of gene length inherently present in EST libraries, gene expression is measured as normalized transcript abundance. It is demonstrated that codon bias and GC content in second codon position are positively associated with transcript abundance. Gene length is negatively associated with transcript abundance. The stability of thermodynamically predicted mRNA secondary structures is not associated with transcript abundance, but there is a negative correlation between mRNA stability and codon bias. This finding does not support the hypothesis that codon bias has evolved as an indirect consequence of selection favoring thermodynamically stable mRNA molecules. PMID- 16044250 TI - Anti-phospholipid antibodies in serum from patients with Guillain-Barre syndrome. AB - OBJECTIVE: The Guillain-Barre syndrome (GBS) is an acute inflammatory polyneuropathy related to autoimmunity. However, no conclusive etiological concept has yet been found. We examined the variation in autoantibodies to lipids in serum of GBS patients in response to the course of the disease but investigated titer modifications during treatment with gamma-globulin. DESIGN AND SETTING: Prospective clinical study in a 14-bed general ICU. PATIENTS: Nine patients with GBS and nine controls were included in the study. MEASUREMENTS AND RESULTS: Four blood samples were obtained before and after treatment. Serum samples, diluted 1:60, were tested by enzyme-linked immunosorbent assay for IgM, IgA, and IgG antibodies to phosphatidylcholine, phosphatidylinositol, cardiolipin, phosphatidic acid, phosphatidylserine, phosphatidylglycerol, phosphatidylethanolamine, sphingomyelin, and gangliosides. Anti-phospholipid antibodies of the IgM, IgA, and IgG families were detected in all GBS patients but in none of the controls. Phosphatidylinositol, cardiolipin, phosphatidylcholine, and phosphatidic acid were the main antigens. All patients developed anti-phosphatidylinositol antibodies of the IgM family and anti cardiolipin antibodies of the IgA and IgG families. A decrease in the level of anti-phospholipid autoantibodies was observed after 1 day of treatment with gamma globulin. Two days after ending gamma-globulin administration the IgG antibodies increased again. CONCLUSIONS: Our findings suggest that in GBS there is an extensive immune reaction, which is altered after gamma-globulin treatment. Anti cardiolipin and anti-phosphatidylinositol antibodies could be useful markers for the response to treatment. PMID- 16044251 TI - Normalizing physiological variables in acute illness: five reasons for caution. PMID- 16044252 TI - Granisetron in the control of radiotherapy-induced nausea and vomiting: a comparison with other antiemetic therapies. AB - Radiotherapy-induced nausea and vomiting (RINV) can be one of the most distressing symptoms of radiotherapy treatment, which if incompletely controlled may last for several weeks with fractionated radiotherapy and prevent completion of the planned treatment course. Current treatment guidelines recommend the use of 5-HT(3) receptor antagonists with or without corticosteroids for highly and moderately emetogenic radiotherapy, though only granisetron and ondansetron are currently indicated for RINV in most countries. Granisetron is a potent and highly selective 5-HT(3) receptor antagonist, with demonstrated efficacy in RINV in both placebo-controlled and comparative studies. In this paper the clinical experience with granisetron in RINV is reviewed, and its efficacy and safety compared with other antiemetic therapies. PMID- 16044253 TI - CpG oligonucleotides enhance the tumor antigen-specific immune response of an anti-idiotype antibody-based vaccine strategy in CEA transgenic mice. AB - A murine monoclonal anti-idiotype (Id) antibody, 3H1 has been developed and characterized previously. Anti-Id 3H1 mimics a specific epitope of carcinoembryonic antigen (CEA) and can be used as a surrogate antigen for CEA. 3H1 induced anti-CEA immunity in different species of animals as well as humans and showed promise as a potential vaccine candidate in phase I/II clinical trials for colon cancer patients. One area of interest to us has been the development of new immune adjuvants that may augment the potency of 3H1 as a tumor vaccine. Oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN) are potent immunostimulatory agents capable of enhancing the Ag-specific Th1 response when used as immune adjuvants. In this study, we have evaluated the efficacy of 3H1 as a tumor vaccine when admixed with a select CpG ODN 1826 in transgenic mice that express human CEA. The vaccine potential of 3H1 was also assessed in the presence of another widely used adjuvant, QS-21. 3H1 coupled to keyhole limpet hemocyanin (KLH) and mixed with Freund's adjuvant (FA) was used as a gold standard in this system. 3H1 vaccination with different adjuvants induced both humoral and cellular anti-3H1, as well as anti-CEA immunity in CEA transgenic mice. The immune sera could lyse CEA-transfected murine colon carcinoma cells, C15 effectively in an antibody-dependent cellular cytotoxicity assay. The anti-CEA antibody responses were somewhat comparable in each adjuvant-treated group of mice, whereas cellular immune responses were significantly greater when CpG was used as an adjuvant. Splenocytes obtained from 3H1-CpG-immunized mice showed an increased proliferative CD4(+) Th1-type T-cell response when stimulated in vitro with 3H1 or CEA and secreted elevated levels of Th1 cytokines (IL-2, IFN-gamma). This vaccine also induced MHC class I antigen-restricted CD8(+) T-cell responses. In a solid tumor model, C15 tumor growth was significantly inhibited by 3H1 vaccinations. In 3H1-CpG-vaccinated mice, the duration of survival was, however, longer compared to the 3H1-QS21-vaccinated mice. These findings suggest that 3H1 CpG vaccinations can break peripheral tolerance to CEA and induce protective antitumor immunity in this murine model transgenic for human CEA. PMID- 16044254 TI - Gene transfer preferentially selects MHC class I positive tumour cells and enhances tumour immunogenicity. AB - The modulated expression of MHC class I on tumour tissue is well documented. Although the effect of MHC class I expression on the tumorigenicity and immunogenicity of MHC class I negative tumour cell lines has been rigorously studied, less is known about the validity of gene transfer and selection in cell lines with a mixed MHC class I phenotype. To address this issue we identified a C26 cell subline that consists of distinct populations of MHC class I (H-2D/K) positive and negative cells. Transient transfection experiments using liposome based transfer showed a lower transgene expression in MHC class I negative cells. In addition, MHC class I negative cells were more sensitive to antibiotic selection. This led to the generation of fully MHC class I positive cell lines. In contrast to C26 cells, all transfectants were rejected in vivo and induced protection against the parental tumour cells in rechallenge experiments. Tumour cell specificity of the immune response was demonstrated in in vitro cytokine secretion and cytotoxicity assays. Transfectants expressing CD40 ligand and hygromycin phosphotransferase were not more immunogenic than cells expressing hygromycin resistance alone. We suggest that the MHC class I positive phenotype of the C26 transfectants had a bearing on their immunogenicity, because selected MHC class I positive cells were more immunogenic than parental C26 cells and could induce specific anti-tumour immune responses. These data demonstrate that the generation of tumour cell transfectants can lead to the selection of subpopulations that show an altered phenotype compared to the parental cell line and display altered immunogenicity independent of selection marker genes or other immune modulatory genes. Our results show the importance of monitoring gene transfer in the whole tumour cell population, especially for the evaluation of in vivo therapies targeted to heterogeneous tumour cell populations. PMID- 16044255 TI - Dynamic control of lymphocyte trafficking by fever-range thermal stress. AB - Migration of blood-borne lymphocytes into tissues involves a tightly orchestrated sequence of adhesion events. Adhesion molecules and chemokine receptors on the surface of circulating lymphocytes initiate contact with specialized endothelial cells under hemodynamic shear prior to extravasation across the vascular barrier into tissues. Lymphocyte-endothelial adhesion occurs preferentially in high endothelial venules (HEV) of peripheral lymphoid organs. The continuous recirculation of naive and central memory lymphocytes across lymph node and Peyer's patch HEV underlies immune surveillance and immune homeostasis. Lymphocyte-endothelial interactions are markedly enhanced in HEV-like vessels of extralymphoid organs during physiological responses associated with acute and chronic inflammation. Similar adhesive mechanisms must be invoked for efficient trafficking of immune effector cells to tumor sites in order for the immune system to have an impact on tumor progression. Here we discuss recent evidence for the role of fever-range thermal stress in promoting lymphocyte-endothelial adhesion and trafficking across HEV in peripheral lymphoid organs. Findings are also presented that support the hypothesis that lymphocyte-endothelial interactions are limited within tumor microenvironments. Further understanding of the molecular mechanisms that dynamically promote lymphocyte trafficking in HEV may provide the basis for novel approaches to improve recruitment of immune effector cells to tumor sites. PMID- 16044256 TI - The effectiveness of montelukast for the treatment of anti-histamine-resistant chronic urticaria. AB - Many patients with chronic idiopathic urticaria are not sufficiently controlled with histamine H(1)-receptor antagonists. Leukotriene receptor antagonists have been reported to be effective for certain cases of urticaria, although their proper application remains to be established. To study the effectiveness of montelukast, a leukotriene receptor antagonist, for the treatment of chronic urticaria that was not controlled by histamine H(1)-receptor antagonists. Twenty five patients with chronic idiopathic urticaria were treated with 10 mg of montelukast for one week or more, without changing any precedent treatment that they were using before the study including histamine H(1)-receptor antagonists. The effectiveness of montelukast for each patient was evaluated and compared with clinical features and/or backgrounds of the patients. Twelve patients, including six who had been treated with corticosteroids, were evaluated as "markedly improved" or "improved" following treatment with montelukast. There was no statistically significant relation of the effectiveness to the complications with non-steroidal anti-inflammatory drugs (NSAIDs) intolerance, mechanical urticaria, or reactions to autologous serum skin test. However, the patients for whom montelukast was effective were younger (33.2+/-16.3 years, mean +/- SD)(P<0.05, Mann-Whitney test) and their duration of illness shorter (15.9+/-18.3 months) (P<0.005, Mann-Whitney test) than those of patients for whom montelukast was ineffective (45.9+/-15.0 years, 89.6+/-71.7 months). Montelukast may be worth trying for patients with chronic idiopathic urticaria, when the condition is not sufficiently controlled with histamine H(1)-receptor antagonists. PMID- 16044257 TI - An insertion/deletion polymorphism in the gene encoding angiotensin converting enzyme is not associated with generalised vitiligo in an English population. AB - Vitiligo is an acquired hypomelanotic skin disorder characterised by circumscribed depigmented macules resulting from the loss of functional melanocytes from the cutaneous epidermis and autoimmunity has been suggested to play a role in the pathogenesis of the disease. Recently, an insertion/deletion (I/D) polymorphism of a 287-base pair repetitive sequence in intron 16 of the angiotensin converting enzyme (ACE) gene has been associated with autoimmune disease and with the development of vitiligo. In this study, the distribution of ACE gene I/D genotypes was investigated in a population of 106 English patients with generalised (non-segmental) vitiligo and 174 ethnically matched healthy controls using a restriction fragment length polymorphism-polymerase chain reaction genotyping method. No significant difference in the frequencies of II, ID and DD genotypes was detected between vitiligo patients and control subjects (P=0.35). The same result was evident for the genotype distribution in vitiligo patients with an autoimmune disease and for those without when compared with controls (P=0.33 and P=0.53, respectively). In addition, the results indicated that the D allele was not significantly over-represented in the group of patients with vitiligo compared with controls (P=0.42) and that this was also the case for patients with and without associated autoimmunity (P=0.40 and P=0.62, respectively). PMID- 16044258 TI - TGF-beta1 up-regulates transglutaminase two and fibronectin in dermal fibroblasts: a possible mechanism for the stabilization of tissue inflammation. AB - Transglutaminase (TGase) has been reported to stabilize tissue inflammation via the mediation of the polymerization of extracellular matrix proteins. A set of cytokines has been implicated in wound healing processes in the dermis. This study was undertaken in order to evaluate the effects of these cytokines on the expression of TGase 2 in human dermal fibroblasts (hDFs), in that TGase 2 is known to be the principal TGase in the dermis. In Western blot analysis, TGF beta1 (1 ng/ml) treatment was found to steadily up-regulate TGase 2 expression for up to 7 days. However, such increases were not observed when the cells were treated with IL-1beta, IL-2, and TNF-alpha. In the enzyme assay, total TGase activities were closely related to the levels of TGase 2 expression. TGase 2 mRNA expression was up-regulated as the result of TGF-beta treatment in competitive RT PCR. In the denatured SDS-PAGE, TGF-beta1 treatment resulted in marked induction of an approximately 220 kDa protein, which was revealed to be a fibronectin (FN) via western immunoblotting with an anti-FN antibody. Next, when the hDFs were treated with TGF-beta1 (1 ng/ml), FN expression was induced beginning at the third day after treatment. The immunoprecipitants generated by anti-FN antibody were positive for the anti-TGase 2 antibody, and the immune complexes were identified at molecular weights of 92 kDa. Collectively, TGF-beta1 stimulates the polymerization of FN via the action of TGase 2, which is supposed to to be an important mechanism in the stabilization of the inflammatory dermis. PMID- 16044259 TI - Identification of genes specific to "oval cells" in the rat 2 acetylaminofluorene/partial hepatectomy model. AB - Under certain conditions liver regeneration can be accomplished by hepatic progenitor cells ("oval cells"). So far, only few factors have been identified to be uniquely regulated by the "oval cell" compartment. Using macroarray analysis in a rat model of oval cell proliferation (treatment with 2-acetylaminofluorene and partial hepatectomy, AAF + PH), we identified 12 differentially expressed genes compared to appropriate control models (AAF treatment and sham operation or AAF treatment alone). Further analysis in models of normal liver regeneration (ordinary PH) and acute phase response (turpentine oil-treated rats) revealed that three out of 12 genes (thymidine kinase 1, Jun-D and ADP-ribosylation factor 4) were not affected by the hepatic acute phase reaction but similarly overexpressed in both "oval cell"-dependant and normal liver regeneration. We characterized Jun-D and ADP-ribosylation factors as novel factors upregulated in oval cells and in non-parenchymal liver cells of normally regenerating livers. However, two out of 12 differentially expressed genes were specifically expressed in oval cells: ras-related protein Rab-3b and Ear-2. On protein level, Rab-3b was increased in total liver homogenates and demonstrated only in clusters of oval cells. We postulate that Ear-2 and Rab-3b may represent novel regulatory factors specifically activated in "oval cells". PMID- 16044260 TI - Coronal and apical lesions, environmental factors: study in a modern and an archeological population. AB - Apical periodontitis (AP) are frequent findings in contemporary dental practice in association with dental pathology or dental care. They have also been studied from an anthropological background. The purpose of this study was to compare the prevalence of apical and dental lesions in an archeological Middle Ages sample and a modern population, and to evaluate the influence of environmental factors. Both the archaeological sample group and dental practice subjects were from southern France. The study included full mouth surveys of 252 individuals (2,780 teeth) from a historic necropolis and 223 subjects (5,678 teeth) randomly selected from the Gard area. Tooth wear, caries, and AP were accounted for clinically and radiographically according to specific indexes. Significant differences were found between period and age in the archeological sample as regards the main risk factors for AP. Antemortem teeth loss and dental wear had been reduced, whereas caries rates and AP had increased between archaeological and modern population. The AP ratio was associated with the level of dental care in the modern population. Although significant variations could be observed between archaeological periods, the rupture in E3 (sixteenth and seventeenth centuries) leads to consider the associated population as a premodern. However, it was found that although cultural and alimentary factors seemed to be the main risk factors in an archeological population, dental care seemed to have a strong influence on AP ratio in modern ones. PMID- 16044261 TI - Primary versus delayed wound closure in complicated appendicitis: an international systematic review and meta-analysis. AB - The purpose of this study was to determine, by means of a systematic review, whether the method of wound closure in complicated appendicitis affects the incidence of wound infection. A comprehensive literature search of multiple databases including MEDLINE (1980-2003), was performed, using the Cochrane search strategy, for articles on wound closure and complicated appendicitis. Clinical trials examining the method of wound closure were selected for systematic review and all quasi-randomized and randomized trials underwent meta-analysis. Failure to close the wound as planned in delayed closure (DC) was considered indicative of a wound infection. Purulent drainage requiring wound opening indicated an infection in the wounds closed primarily. Six randomized trials were considered adequate for meta-analysis. None independently showed a statistically significant difference in the risk of developing a wound infection with primary closure (PC). When pooled data were subjected to meta-analysis, PC achieved a statistically significant reduction in the relative risk of treatment failure and did not lead to an increase in wound infections. Primary closure does not increase the risk of developing a wound infection after operation for perforated appendicitis. Given the lack of benefit of DC, and the less traumatic, less painful, and less costly nature of PC; primary closure is a safe and practical treatment option. PMID- 16044262 TI - Gene cloning and functional analysis of glycosaminoglycan-degrading enzyme chondroitin AC lyase from Flavobacterium columnare G4. AB - The chondroitin AC lyase gene, cslA, was cloned for the first time from the fish bacterial pathogen F. columnare G4. From the first transcription initiation site, the cslA extends 2620 nucleotides to the end of the 3' region. The open reading frame of cslA transcript has 2286 nucleotides encoding 762 amino acids with a 16 residues long signal peptide at the N-terminus. The gene, cslA was then successfully expressed in Escherichia coli and recombinant chondroitin AC lyase, rChonAC was purified, with its lytic activity analyzed. Zymography analysis copolymerized with chondroitin sulphate revealed the lytic activity of rChonAC and also the crude native ChonAC isolated from periplamic space of cultured F. columnare G4. The low level of lytic activity observed in crude native ChonAC may be due possibly to the low level of expression of this gene in the cultured condition. The expression and the role of this virulence factor is of interest for further research on the pathogenesis of F. columnare. PMID- 16044263 TI - A single operon-encoded form of the acetyl-CoA decarbonylase/synthase multienzyme complex responsible for synthesis and cleavage of acetyl-CoA in Methanosarcina thermophila. AB - Methanogens growing on C-1 substrates synthesize 2-carbon acetyl groups in the form of acetyl-CoA for carbon assimilation using the multienzyme complex acetyl CoA decarbonylase/synthase (ACDS) which contains five different subunits encoded within an operon. In species growing on acetate ACDS also functions to cleave the acetate C-C bond for energy production by methanogenesis. A number of species of Methanosarcina that are capable of growth on either C-1 compounds or acetate contain two separate ACDS operons, and questions have been raised about whether or not these operons play separate roles in acetate synthesis and cleavage. Methanosarcina thermophila genomic DNA was analyzed for the presence of two ACDS operons by PCR amplifications with different primer pairs, restriction enzyme analyses, DNA sequencing and Southern blot analyses. A single ACDS operon was identified and characterized, with no evidence for more than one. MALDI mass spectrometric analyses were carried out on ACDS preparations from methanol- and acetate-grown cells. Peptide fragmentation patterns showed that the same ACDS subunits were present regardless of growth conditions. The evidence indicates that a single form of ACDS is used both for acetate cleavage during growth on acetate and for acetate synthesis during growth on C-1 substrates. PMID- 16044264 TI - Transcriptional response reveals translation machinery as target for high pressure in Lactobacillus sanfranciscensis. AB - The effect of sublethal hydrostatic pressure on the transcriptome of Lactobacillus sanfranciscensis was determined using a shot-gun-microarray. Among the 750 spots that passed quality analysis 42 genes were induced, while six were repressed when cells were incubated at 45 MPa for 30 min. The nature of genes and their differential expression clearly indicate cellular efforts to counteract a decrease in translational capacity. The majority of high pressure affected genes were found to encode either translation factors (EF-G, EF-TU), ribosomal proteins (S2, L6, L11), genes changing translational accuracy or molecular chaperones (GroEL, ClpL). These data agree with previously reported effects observed in in vitro studies as well as with physiological and proteomic data. This study provides in vivo evidence to identify ribosomes and impaired translation among primary targets for high pressure treatment. The observed induction of heat as well as cold shock genes (e.g. hsp60, gyrA) may be explained as a result of high pressure affected protein synthesis. PMID- 16044265 TI - Topological and transcriptional analysis of pssL gene product: a putative Wzx like exopolysaccharide translocase in Rhizobium leguminosarum bv. trifolii TA1. AB - An identified pssL gene is yet another one, besides the pssT, pssN and pssP genes, encoding for a protein engaged in polysaccharide polymerization and export in Rhizobium leguminosarum bv. trifolii strain TA1 (RtTA1). Amino acid sequence similarity and hypothetical protein secondary structure placed the PssL protein within Wzx (RfbX) translocases with putative flippase function that belong to the polysaccharide specific transport (PST) family. The predicted secondary structure of the PssL membrane protein was examined with a series of PssL-PhoA and PssL LacZ translational fusions. The results support the hypothesis of PssL being a member of PST protein family comprising transporters with 12 membrane spanning segments and amino and carboxyl termini located in the cytoplasm. Results of semi quantitative RT-PCR showed that the initial abundance of mRNA encoding PssL protein was relatively lower when compared to the quantity of the previously identified PssT membrane protein. PssL might be a good candidate for Wzx-like protein that together with PssT (Wzy protein) could be responsible for Wzx/Wzy like-dependent EPS polymerization and translocation in RtTA1. PMID- 16044266 TI - Genetic mapping of a cross between Gossypium hirsutum (cotton) and the Hawaiian endemic, Gossypium tomentosum. AB - The existence of five tetraploid species that derive from a common polyploidization event about 1 million years ago makes Gossypium (cotton) an attractive genus in which to study polyploid evolution and offers opportunities for crop improvement through introgression. To date, only crosses (HB) between the cultivated tetraploid cottons Gossypium hirsutum and G. barbadense have been genetically mapped. Genetic analysis of a cross (HT) between G. hirsutum and the Hawaiian endemic G. tomentosum is reported here. Overall, chromosomal lengths are closely correlated between the HB and HT maps, although there is generally more recombination in HT, consistent with a closer relationship between the two species. Interspecific differences in local recombination rates are observed, perhaps involving a number of possible factors. Our data corroborate cytogenetic evidence that chromosome arm translocations have not played a role in the divergence of polyploid cottons. However, one terminal inversion on chromosome (chr.) 3 does appear to differentiate G. tomentosum from G. barbadense; a few other apparent differences in marker order fall near gaps in the HT map and/or lack the suppression of recombination expected of inversions, and thus remain uncertain. Genetic analysis of a discrete trait that is characteristic of G. tomentosum, nectarilessness, mapped not to the classically reported location on chr. 12 but to the homoeologous location on chr. 26. We propose some hypotheses for further study to explore this incongruity. Preliminary quantitative trait locus (QTL) analysis of this small population, albeit with a high probability of false negatives, suggests a different genetic control of leaf morphology in HT than in HB, which also warrants further investigation. PMID- 16044267 TI - Comparative analysis of a transposon-rich Brassica oleracea BAC clone with its corresponding sequence in A. thaliana. AB - We compared the sequence of a 96.7 Kb-long BAC clone (B 19 N 3) from Brassica oleracea (broccoli) with its corresponding regions in Arabidopsis thaliana. B 19 N 3 contains eight genes and 15 transposable elements (TEs). The first two genes in this clone, Bo 1 and Bo 2, have its corresponding region at the end of chromosome V of Arabidopsis (24 Mb). The third gene, Bo 3, corresponds to an ortholog at the opposite end (2.6 Mb) of the same chromosome. The other five genes, Bo 4 to Bo 8 also have a corresponding region on the same chromosome but at 7.7 Mb . These five genes are colinear with those found in the corresponding region of Arabidopsis, which contains, however, 15 genes. Therefore, a cluster of 10 genes is missing in B. oleracea clone (B 19 N 3). All five genes in common have the same order and orientation in the genomes of both species. Their 36 exons constituting the eight homologous genes have high conservation in size and sequence identity in both species. Among these, there is a major gene involved in aliphatic glucosinolate biosynthesis, Bo GSL-ELONG (Bo 4). Similar to A. thaliana, this gene, has a tandem duplicate, Bo 5. A contig for this region was constructed by primer walking and BAC-end-sequencing, revealing general gene colinearity between both species. During the 20 million years separating A. thaliana from B. oleracea from a common ancestor both genomes have diverged by chromosomal rearrangements and differential TE activity. These events, in addition to changes in chromosome number are responsible for the evolution of the genomes of both species. In spite of these changes, both species conserve general colinearity for their corresponding genes. PMID- 16044268 TI - Development and characterization of wheat- Leymus racemosus translocation lines with resistance to Fusarium Head Blight. AB - Wheat scab (Fusarium Head Blight, FHB) is a destructive disease in the warm and humid wheat-growing areas of the world. Finding diverse sources of FHB resistance is critical for genetic diversity of resistance for wheat breeding programs. Leymus racemosus is a wild perennial relative of wheat and is highly resistant to FHB. Three wheat- L. racemosus disomic addition (DA) lines DA 5 Lr#1, DA 7 Lr#1 and DALr.7 resistant to FHB were used to develop wheat- L.racemosus translocation lines through irradiation and gametocidal gene-induced chromosome breakage. A total of nine wheat-alien translocation lines with wheat scab resistance were identified by chromosome C-banding, GISH, telosomic pairing and RFLP analyses. In line NAU 614, the long arm of 5 Lr#1 was translocated to wheat chromosome 6B. Four lines, NAU 601, NAU 615, NAU 617, and NAU 635, had a part of the short arm of 7 Lr#1 transferred to different wheat chromosomes. Four other lines, NAU 611, NAU 634, NAU 633, and NAU 618, contained translocations involving Leymus chromosome Lr.7 and different wheat chromosomes. The resistance level of the translocation lines with a single alien chromosome segment was higher than the susceptible wheat parent Chinese Spring but lower than the alien resistant parent L. racemosus. At least three resistance genes in L. racemosus were identified. One was located on chromosome Lr.7, and two could be assigned to the long arm of 5 Lr#1 and the short arm of 7 Lr#1. PMID- 16044269 TI - Cloning of resistance gene analogs located on the alien chromosome in an addition line of wheat-Thinopyrum intermedium. AB - Homology-based gene/gene-analog cloning method has been extensively applied in isolation of RGAs (resistance gene analogs) in various plant species. However, serious interference of sequences on homoeologous chromosomes in polyploidy species usually occurred when cloning RGAs in a specific chromosome. In this research, the techniques of chromosome microdissection combined with homology based cloning were used to clone RGAs from a specific chromosome of Wheat Thinopyrum alien addition line TAi-27, which was derived from common wheat and Thinopyrum intermedium with a pair of chromosomes from Th. intermedium. The alien chromosomes carry genes for resistance to BYDV. The alien chromosome in TAi-27 was isolated by a glass needle and digested with proteinase K. The DNA of the alien chromosome was amplified by two rounds of Sau3A linker adaptor-mediated PCR. RGAs were amplified by PCR with the degenerated primers designed based on conserved domains of published resistance genes (R genes) by using the alien chromosome DNA, genomic DNA and cDNA of Th. intermedium, TAi-27 and 3B-2 (a parent of TAi-27) as templates. A total of seven RGAs were obtained and sequenced. Of which, a constitutively expressed single-copy NBS-LRR type RGA ACR 3 was amplified from the dissected alien chromosome of TAi-27, TcDR 2 and TcDR 3 were from cDNA of Th. intermedium, AcDR 3 was from cDNA of TAi-27, FcDR 2 was from cDNA of 3B-2, AR 2 was from genomic DNA of TAi-27 and TR 2 was from genomic DNA of Th. intermedium. Sequence homology analyses showed that the above RGAs were highly homologous with known resistance genes or resistance gene analogs and belonged to NBS-LRR type of R genes. ACR 3 was recovered by PCR from genomic DNA and cDNA of Th. intermedium and TAi-27, but not from 3B-2. Southern hybridization using the digested genomic DNA of Th. intermedium, TAi-27 and 3B-2 as the template and ACR 3 as the probe showed that there is only one copy of ACR 3 in the genome of Th. intermedium and TAi-27, but it is absent in 3B-2. The ACR 3 could be used as a specific probe of the R gene on the alien chromosome of TAi 27. Results of Northern hybridization suggested that ACR 3 was constitutively expressed in Th. intermedium and TAi-27, but not 3B-2, and expressed higher in leaves than in roots. This research demonstrated a new way to clone RGAs located on a specific chromosome. The information reported here should be useful to understand the resistance mechanism of, and to clone resistant genes from, the alien chromosome in TAi-27. PMID- 16044270 TI - Mapping of extreme resistance to PVY (Ry (sto)) on chromosome XII using anther culture-derived primary dihaploid potato lines. AB - The inheritance of extreme resistance to PVY (Ry (sto)) by a single dominant locus was confirmed by obtaining a 1:1 segregation ratio in a virus inoculation test with 28 resistant (Ryry) to 29 susceptible (ryry) anther culture-derived dihaploid lines (2n=2x=24) from cv. "Assia" (2n=4x=48) having extreme resistance derived from Solanum stoloniferum in simplex constitution (Ryryryry). Twelve Ry (sto) markers selected in AFLP assays using bulked segregant analysis were applied to 106 tested potato cultivars from Germany, The Netherlands and Poland and 19 potato cultivars were identified by these markers as extremely resistant to PVY in alignment with phenotypic data. The locus for extreme resistance (Ry (sto)) to PVY was mapped on chromosome XII co-segregating with the SSR marker STM 0003. The utility of anther-culture derived dihaploid potatoes for genetic marker development was demonstrated. Marker transferability from diploids to tetraploids provides an optimistic potential for marker-assisted selection in potato breeding programs. PMID- 16044271 TI - [Chronic ulceration of one side of the face]. PMID- 16044272 TI - [Tasks and function of the German Medical Review Board of the Statutory Health Insurance (MDK). Latest aspects in dermatology in view of social medicine]. AB - The German Medical Review Board of the Statutory Health Insurance (MDK) is a sociomedical service for impartial medical opinion and consultation. Together with the statutory health insurance companies it takes part in shaping the public health care system. Many recent developments are changing the german health insurance system. This article illustrates the complex sociomedical framework of MDK's activities in relation to current issues in dermatology. The article focuses on: new diagnostic and therapeutic methods, drugs (off-label-use), out patient and in-patient dermatological rehabilitation and hospital treatment. PMID- 16044274 TI - Fulminant hepatic failure in a newborn with herpes simplex virus 2 infection. PMID- 16044275 TI - Atypical manifestation of childhood primary cerebral lymphoma restricted to the leptomeninges. PMID- 16044273 TI - Ion channel clustering enhances weak electric field detection by neutrophils: apparent roles of SKF96365-sensitive cation channels and myeloperoxidase trafficking in cellular responses. AB - We have tested Galvanovskis and Sandblom's prediction that ion channel clustering enhances weak electric field detection by cells as well as how the elicited signals couple to metabolic alterations. Electric field application was timed to coincide with certain known intracellular chemical oscillators (phase-matched conditions). Polarized, but not spherical, neutrophils labeled with anti-K(v)1.3, FL-DHP, and anti-TRP1, but not anti-T-type Ca(2+) channels, displayed clusters at the lamellipodium. Resonance energy transfer experiments showed that these channel pairs were in close proximity. Dose-field sensitivity studies of channel blockers suggested that K(+) and Ca(2+) channels participate in field detection, as judged by enhanced oscillatory NAD(P)H amplitudes. Further studies suggested that K(+) channel blockers act by reducing the neutrophil's membrane potential. Mibefradil and SKF93635, which block T-type Ca(2+) channels and SOCs, respectively, affected field detection at appropriate doses. Microfluorometry and high-speed imaging of indo-1-labeled neutrophils was used to examine Ca(2+) signaling. Electric fields enhanced Ca(2+) spike amplitude and triggered formation of a second traveling Ca(2+) wave. Mibefradil blocked Ca(2+) spikes and waves. Although 10 microM SKF96365 mimicked mibefradil, 7 microM SKF96365 specifically inhibited electric field-induced Ca(2+) signals, suggesting that one SKF96365-senstive site is influenced by electric fields. Although cells remained morphologically polarized, ion channel clusters at the lamellipodium and electric field sensitivity were inhibited by methyl-beta-cyclodextrin. As a result of phase-matched electric field application in the presence of ion channel clusters, myeloperoxidase (MPO) was found to traffic to the cell surface. As MPO participates in high amplitude metabolic oscillations, this suggests a link between the signaling apparatus and metabolic changes. Furthermore, electric field effects could be blocked by MPO inhibition or removal while certain electric field effects were mimicked by the addition of MPO to untreated cells. Therefore, channel clustering plays an important role in electric field detection and downstream responses of morphologically polarized neutrophils. In addition to providing new mechanistic insights concerning electric field interactions with cells, our work suggests novel methods to remotely manipulate physiological pathways. PMID- 16044276 TI - Long-term serial echocardiographic examination of late anthracycline cardiotoxicity and its prevention by dexrazoxane in paediatric patients. AB - The authors conducted an 8-year prospective non-randomised study to determine whether dexrazoxane (ICRF-187) would reduce late anthracycline-induced cardiotoxicity in patients treated in childhood for haematological malignancy. The authors examined prospectively 75 patients (40 male/35 female) aged 2-17 years (median 6.5 years) at the time of diagnosis. The cardioprotection was given to 53 patients (26 male/17 female) and the standard protocol was used in 22 patients (14 male/8 female). The prospective echocardiographic evaluation was done before and after the chemotherapy and every year during the follow-up period. Dynamic stress echocardiography (DSE) was assessed in the final year. The clinical cardiotoxicity was not diagnosed. Higher cumulative doses of anthracycline were given in the dexrazoxane group (234+/-58 mg/m(2), median 240 mg/m(2) versus 203+/-86 mg/m(2), median 210 mg/m(2), P <0.04) and a significantly higher percentage of patients received cumulative doses >240 mg/m(2) of anthracycline ( P <0.05). During the follow-up period, the fractional shortening (FS) declined in the no-dexrazoxane group only in the 8th year and was significantly lower compared to the dexrazoxane group ( P <0.05). The pathological decrease in FS was present in 24% of patients; 41% in the no dexrazoxane and 17% in the dexrazoxane groups, respectively ( P <0.05). DSE demonstrated lower rest EF and cardiac index (CI) in the no-dexrazoxane group ( P <0.05); however, neither the response of EF and CI to the stress echocardiography nor the exercise tolerance significantly differed between sub-groups. A higher number of patients in the dexrazoxane group had very good exercise tolerance (ET) >3 Watts/kg ( P <0.05) and a lower number responded with a decreased ET <2 Watts/kg ( P <0.05) compared to the no-dexrazoxane group. CONCLUSION: Dexrazoxane seems to reduce the risk of late subclinical cardiotoxicity. Dexrazoxane-treated patients revealed better exercise tolerance; however the haemodynamic response to the stress was no different in both sub-groups. PMID- 16044277 TI - Reduced aortic elastic properties in a child with Takayasu arteritis: case report and literature review. AB - Takayasu arteritis (TA) is a chronic inflammatory vasculitis of the aorta and its major branches with a very low incidence in Europe and North America. Our objective was to determine the elastic properties of the affected ascending and descending aortic walls non-invasively in a 14-year-old Iraqi girl with a 3-year history of fever, fatigue, malaise and diffuse pain. Ultrasound and magnetic resonance angiography showed marked thickening of the aortic wall, dilatation of the aortic arch, and decreased luminal diameters of the abdominal aorta and both subclavian arteries, consistent with TA. Ascending and descending aortic elastic properties such as distensibility and stiffness index were markedly reduced compared to a group of healthy controls (n=39): ascending aortic distensibility was 20 kPa(-1) x 10(-3) versus 63+/-23 kPa(-1) x 10(-3) in controls, and the ascending aortic stiffness index 9.6 versus 3.5+/-1.3 in controls. Although the patient's general condition improved rapidly on oral prednisolone and azathioprine and inflammatory parameters normalised within 3 weeks, the aortic elastic parameters did not change during the first 2 weeks of anti-inflammatory treatment. Unfortunately, no further follow-up was possible. CONCLUSION: In patients with Takayasu arteritis, non-invasive quantification of reduced aortic elastic properties can help to assess aortic involvement, and possibly to follow disease activity and vascular response to therapy. PMID- 16044278 TI - Nineteen cases of persistent pruritic nodules and contact allergy to aluminium after injection of commonly used aluminium-adsorbed vaccines. AB - Rare cases of persistent pruritic nodules, sometimes associated with aluminium (Al) allergy, have been reported after the use of several Al adsorbed vaccines. During vaccine trials in the 1990s a high incidence of pruritic nodules (645 cases/76,000 recipients), in 77% associated with Al allergy, was observed after the administration of diphtheria-tetanus / acellular pertussis (DT/aP) vaccines from a single producer. In the present report 19 children with pruritic nodules after vaccination with Al hydroxide-adsorbed DTaP/polio+Hib (Infanrix, Pentavac) are described. The children had intensely itching nodules at the injection site, often aggravated during upper respiratory tract infections, and local skin alterations. So far, the symptoms have persisted for up to 7 years. The median time between vaccination and onset of symptoms was 1 month. 16 children were epicutaneously tested for Al, all with positive reactions indicating delayed hypersensitivity to Al. The condition is not commonly known but is important to recognise, as the child and the family may suffer considerably. Future vaccinations with Al-adsorbed vaccines may cause aggravation of the symptoms and the Al allergy. Al-containing skin products, such as antiperspirants, may cause contact dermatitis. Nodules may be mistaken for tumours. Even though the incidence of itching nodules and Al allergy after administration of Infanrix, Pentavac and other Al-adsorbed vaccines is probably low, research to replace Al adjuvants seems appropriate. We conclude that intensely itching subcutaneous nodules, lasting for many years, and hypersensitivity to aluminium may occur after DTaP/polio+Hib vaccination of infants. PMID- 16044279 TI - A case of severe Pierre Robin sequence with failure to thrive and tachycardia resolved after redo-fundoplication and hiatoplasty. AB - We report an infant suffering from Pierre Robin sequence complicated by gastro oesophageal reflux and failure to thrive, which were resistant to conservative therapy and a hemifundoplication. Gastro-oesophageal reflux was accompanied by supraventricular tachycardia, treated with propafenone. Tachycardia may be present in Pierre Robin sequence as a consequence of cardiac parasympathetic imbalance. The patient recovered completely from the gastro-oesophageal reflux and tachycardia after redo-fundoplication (Nissen) and a hiatoplasty were performed. CONCLUSION: This case shows that a thorough search for gastro oesophageal reflux is indicated in each case of Pierre Robin sequence with failure to thrive. PMID- 16044280 TI - [Conventional techniques of visual field examination Part 2: confrontation visual field testing -- kinetic perimetry]. AB - Evaluation of visual field findings is an essential ophthalmological task. Applying psychophysical principles enables noninvasive (topo-)diagnosis in cases of unexplained visual loss or suspected lesions of the visual pathway. Furthermore, any kind of expert opinion or ability test critically depends on adequate visual field findings. Repeated examinations allow functional follow-up of a disease and assessment of therapeutic efficiency. This paper explicitly focuses on conventional strategies since these are widespread, easily accessible, standardized, and well established in everyday practice over years. Standardized white-on-white perimetry with moving targets (kinetic perimetry) is the major topic of this paper (part 2) in addition to "rough"/preliminary confrontation visual field testing. This review series is aimed at imparting basic concepts in perimetry and psychophysics to ophthalmologists and also addresses interested physicians from other disciplines. PMID- 16044281 TI - Functional characterization of the Candida albicans CRZ1 gene encoding a calcineurin-regulated transcription factor. AB - Calcineurin is a phosphoprotein phosphatase devoted to the transduction of Ca(2+) signals in eukaryotes. In the human pathogen Candida albicans, calcineurin function is required for cell morphogenesis, azole tolerance, membrane stress responses, survival in serum and virulence in mice. Molecular mechanisms as well as targets downstream C. albicans calcineurin involved in all these processes are still uncharacterized. Here we report the characterization of a C. albicans Crz1 calcineurin-regulated transcription factor using a Saccharomyces cerevisiae crz1Delta defective strain as heterologous host. CaCrz1p fulfils the function of its S. cerevisiae homolog protein to control the expression of several Ca(2+)/calcineurin-responsive genes acting on the CDRE sequence in promoters. In the model yeast, CaCrz1p activity and localization are regulated by calcineurin. Deletion of CRZ1 gene renders C. albicans hypersensitive to alkaline cations and membrane stress conditions, including that elicited by SDS and antifungal azoles. Our findings indicate that CaCrz1p is member of a calcium-regulated pathway required for the maintenance of membrane integrity. PMID- 16044282 TI - [Aftercare of urological tumors---why and how?]. PMID- 16044283 TI - [Assessment among German urologists of various conservative treatment modalities for Peyronie's disease. Results of a survey]. AB - BACKGROUND: The aim of this study was to evaluate the frequency and the assessment of the different conservative modalities of treatment in Peyronie's disease. A representative survey among 3187 German urologists was performed using a standardized questionnaire comprising currently used concepts of therapy, their efficacy, and their tolerability. MATERIAL AND METHODS: A total of 636 urologists participated in the study. Altogether they had treated 6019 patients with Peyronie's disease in 2003. The majority of urologists treated between 3 and 15 patients per year. The most frequent treatment modality was the administration of potassium paraaminobenzoate, followed by vitamin E and extracorporeal shock wave therapy. Other oral drugs and intralesional drug administrations were used considerably less frequently. RESULTS: The most commonly used treatment modalities were assessed for the best results in terms of efficacy and tolerability. However, this outcome is contradictory to the few controlled studies regarding the efficacy of the different drugs. CONCLUSIONS: The large number of patients treated demonstrates the importance of conservative therapy for Peyronie's disease. Therefore, it is unfortunate that no conservative treatment modality is currently available that cures the symptoms of this disorder in all patients affected. PMID- 16044284 TI - Modelling and optimisation of enzymatic separating micro-reactor. AB - A mathematical model of an enzymatic separating microreactor with the electro osmotic control of reaction component transport rates is analysed. The micro reactor is considered in a form of a thin channel filled with a gel containing an immobilised enzyme and an adsorbent where the enzyme reaction, the molecular diffusion, the electro-osmotic flux and the adsorption take place. The substrate inhibited enzyme reaction splitting a non-ionic substrate to two non-ionic products is considered. The reactor operates in a periodic regime, when the channel entry is exposed to the periodic substrate concentration pulses. A chromatographic separation of reaction components, therefore, proceeds in the channel. Effects of principal operational parameters of the reactor system-the reaction channel length, the electric current density, the substrate inlet concentration, the rate of adsorption, and the enzyme activity--on resolution of the products at reactor outlet are analysed. The existence of optimum parameter values (maximising the resolution of reaction products) is shown and a multiparametric optimisation of the reactor performance is accomplished. PMID- 16044285 TI - Polyhydric alcohol protective effect on Rhizomucor miehei lipase deactivation enhanced by pressure and temperature treatment. AB - The influence of polyhydric alcohols (sorbitol, xylitol, erythritol, glycerol) on the thermal stability of Rhizomucor miehei lipase has been studied at high hydrostatic pressure (up to 500 MPa). In the absence of additives, a protective effect (PE) (the ratio between the residual activities determined at 480 MPa for the enzyme in the presence or absence of polyhydric alcohols) of low-applied pressures (from 50 MPa to 350 MPa) against thermal deactivations (at 50 degrees C and 55 degrees C) has been noticed. In the presence of additives, a strong correlation between PE and the total hydroxyl group concentration has been obtained, for the first time, under treatments of combining denaturing temperatures and high hydrostatic pressures. This relationship does not seem to be dependent on the nature polyhydric alcohols as the same effect could be observed with 1 M sorbitol and 2 M glycerol. This PE, against thermal and high pressure combined lipase deactivation, increases with polyhydric alcohol concentrations, and when temperature increases from 25 degrees C to 55 degrees C. PMID- 16044286 TI - Evaluation of the effects and interactions of mixing and oxygen transfer on the production of Fab' antibody fragments in Escherichia coli fermentation with gas blending. AB - Fermentations carried out at 450-L and 20-L scale to produce Fab' antibody fragments indicated a serious problem to control levels of dissolved oxygen in the broth due to the large oxygen demand at high cell densities. Dissolved oxygen tension (DOT) dropped to zero during the induction phase and it was hypothesised that this could limit product formation due to inadequate oxygen supply. A gas blending system at 20-L scale was employed to address this problem and a factorial 2(2) experimental design was executed to evaluate independently the effects and interaction of two main engineering factors: agitation rate and DOT level (both related to mixing and oxygen transfer in the broth) on Fab' yields. By comparison to the non-gas blending system, results in the gas blending system at same scale showed an increase in the production of Fab' by 77% independent of the DOT level when using an agitation rate of 500 rpm level and by 50% at an agitation rate of 1,000 rpm with 30% DOT. Product localisation in the cell periplasm of >90% was obtained in all fermentations. Results obtained encourage further studies at 450-L scale initially, to evaluate the potential of gas blending for the industrial production of Fab' antibody fragments. PMID- 16044287 TI - Study of the inhibitory effect of the product dihydroxyacetone on Gluconobacter oxydans in a semi-continuous two-stage repeated-fed-batch process. AB - The influence of the product inhibition by dihydroxyacetone (DHA) on Gluconobacter oxydans for a novel semi-continuous two-stage repeated-fed-batch process was examined quantitatively. It was shown that the culture was able to grow up to a DHA concentration of 80 kg m(-3) without any influence of product inhibition. The regeneration capability of the reversibly product inhibited culture from a laboratory-scale bioreactor system was observed up to a DHA concentration of about 160 kg m(-3). At higher DHA concentrations, the culture was irreversibly product inhibited. However, due to the robust membrane-bound glycerol dehydrogenase of G. oxydans, product formation was still active for a prolonged period of time. The reachable maximum final DHA concentration was as high as 220 kg m(-3). The lag phases for growth increased exponentially with increasing DHA threshold values of the first reactor stage. These results correlated well with fluorescence in situ hybridization (FISH) measurements confirming that the number of active cells decreased exponentially with increasing DHA concentrations. PMID- 16044288 TI - Design of europium(III) complexes with high quantum yield. AB - This work describes a rational planning of a new light-conversion molecular device with high quantum yield. For this, we made modifications in the 3-amino-2 carboxypyridine and 3-amino-2-carboxypirazine acid ligands, generating eight different complexes. Theoretical methods have been used to calculate the quantum yield of each of the complexes. We first used the Sparkle model to calculate the ground-state geometries of the eight complexes. These data were used to perform theoretical predictions of the energy transitions using the INDO/S-CI method. After having obtained the geometry and the energy transitions, the energy transfer rates and quantum yield were calculated using a theoretical approach based on the application of the 4f-4f transition theory. The results show that the modifications in the 3-amino-2-carboxypyridine ligand had generated three complexes with high quantum yield (about 52.8, 51.6 and 52.8%). On the other hand, the modifications in the 3-amino-2-carboxypirazine led to only one complex with quantum yield larger than 50%, but it is the most efficient complex projected. PMID- 16044289 TI - H-bonded complexes between acetylacetone and two molecules of methanol: HF and DFT level study. AB - Five stable H-bonded complexes (supersystems) between acetylacetone and two methanol molecules were investigated at the B3LYP and HF levels of theory using the 6-311G** and 6-11++G** basis sets. The most stable complex was found as the one with the highest relative bonding and interaction energies. All vibrational frequencies resulting from calculations with the 6-311++G** basis set were compared with the recorded IR spectrum of acetylacetone/methanol mixture in a molar ratio 1:2. PMID- 16044290 TI - Antifungal activities of actinomycete strains associated with high-altitude sagebrush rhizosphere. AB - The antifungal-producing potential of actinomycete populations from the rhizosphere of low-altitude sagebrush, Artemisia tridentata, has been examined. In a continued investigation of new sources of antifungal-producing microorganisms, this study examined the antifungal-producing potential of actinomycetes from the rhizosphere of high-altitude A. tridentata. With high altitude sagebrush, rhizosphere soil actinomycete numbers were one to four orders of magnitude higher than those found in nonrhizosphere bulk soils and different from those found with the low-altitude plants. A total of 122 actinomycete isolates was screened against nine fungal species and six bacterial species for the production of antimicrobial compounds. Four rhizosphere isolates, Streptomyces amakusaensis, S. coeruleorubidus, S. hawaiiensis and S. scabies, showed broad-spectrum antifungal activity against three or more fungal species in plate assays. In liquid antagonism assays, mycelium production by Aspergillus niger was reduced by up to 50% by two of the actinomycete isolates. These results demonstrate the potential of rhizosphere microbiology in the search for new antimicrobials. PMID- 16044291 TI - Enhanced photocatalytic inactivation of bacterial spores on surfaces in air. AB - TiO(2) photocatalysis with ultraviolet (UV-A) light has proven to be a highly effective process for complete inactivation of airborne microbes. However, the overall efficiency of the technology needs to be improved to make it more attractive as a defense against bio-terrorism. The present research investigates the enhancement in the rate of destruction of bacterial spores on metal (aluminum) and fabric (polyester) substrates with metal (silver)-doped titanium dioxide and compares it to conventional photocatalysis (TiO(2) P25/+UV-A) and UV A photolysis. Bacillus cereus bacterial spores were used as an index to demonstrate the enhanced disinfection efficiency. The results indicate complete inactivation of B. cereus spores with the enhanced photocatalyst. The enhanced spore destruction rate may be attributed to the highly oxidizing radicals generated by the doped TiO(2). PMID- 16044292 TI - Production of 1,3-Propanediol by Clostridium butyricum VPI 3266 in continuous cultures with high yield and productivity. AB - The effects of dilution rate and substrate feed concentration on continuous glycerol fermentation by Clostridium butyricum VPI 3266, a natural 1,3 propanediol producer, were evaluated in this work. A high and constant 1,3 propanediol yield (around 0.65 mol/mol), close to the theoretical value, was obtained irrespective of substrate feed concentration or dilution rate. Improvement of 1,3-propanediol volumetric productivity was achieved by increasing the dilution rate, at a fixed feed substrate concentration of 30, 60 or 70 g l( 1). Higher 1,3-propanediol final concentrations and volumetric productivities were also obtained when glycerol feed concentration was increased from 30 to 60 g l(-1), at D=0.05-0.3 h(-1), and from 60-70 g l(-1), at D=0.05 and 0.1 h(-1).30 g l(-1) of 1,3-propanediol and the highest reported value of productivity, 10.3 g l(-1) h(-1), was achieved at D=0.30 h(-1) and 60 g l(-1) of feed glycerol. A switch to an acetate/butyrate ratio higher than one was observed for 60 g l(-1) of feed glycerol and a dilution rate higher than 0.10 h(-1); moreover, at D=0.30 h(-1) 3-hydroxypropionaldehyde accumulation was observed for the first time in the fermentation broth of C. butyricum. PMID- 16044293 TI - Biosynthesis of poly-beta-hydroxyalkanoates by Sphingopyxis chilensis S37 and Wautersia sp. PZK cultured in cellulose pulp mill effluents containing 2,4,6 trichlorophenol. AB - Poly-beta-hydroxyalkanoates (PHA) polymer is synthesized by different bacterial species. There has been considerable interest in the development and production of biodegradable polymers; however, the high cost of PHA production has restricted its applications. Kraft cellulose industry effluents containing 2,4,6 trichlorophenol (10 or 20 microg ml(-1)) were used by the bacteria Sphingopyxis chilensis S37 and Wautersia sp. PZK to synthesize PHA. In this condition, S. chilensis S37 was able to grow and degrade 2,4,6-trichlorophenol (ca. 60%) and 80% of these cells accumulated PHA. Wautersia PZK completely degraded 2,4,6-TCP and more than 90% of the cells accumulated PHA in 72 h. The PHA detection was performed by flow cytometry and polyester composition was characterized by gas chromatography-mass spectroscopy (GC-MS), indicating that these polymers are made by 3-hydroxybutyric acid and 3-hydroxyhexadecanoic acid for S37 and PZK strains, respectively. Results demonstrated that strains' growth and PHA production and composition are not modified in cellulose effluents with or without 2,4,6-TCP (10 20 microg ml(-1)). Therefore, our results indicate that S. chilensis S37 and Wautersia sp. PZK are able to degrade a toxic compound such as a 2,4,6-TCP and simultaneously produce a valuable biopolymer using low-value substrates. PMID- 16044294 TI - Biomechanical evaluation of a bioabsorbable expansion bolt for hamstring graft fixation in ACL reconstruction: an experimental study in calf tibial bone. AB - INTRODUCTION: The purpose of our study was to evaluate and compare the primary fixation strength of a novel bioabsorbable two shell expansion bolt (EB) with that of a well-established interference screw-fixation technique in hamstring reconstruction of the anterior cruciate ligament. MATERIALS AND METHODS: Thirty calf tibia plateaus (age 5-6 months) were assigned to three groups: In group I (n = 10) triple-stranded hamstring grafts were fixed with titanium interference screws (7 mm thread / 8 mm head x 25 mm). Specimens of group II (n = 10) received bioabsorbable poly-L-lactide interference screws (8 x 23 mm). In group III (n = 10), the grafts were fixed using bioabsorbable poly-D,L-lactide expansion bolts (5.8/8.5/10 mm x 35 mm). The tensile axis was placed parallel to the bone tunnel. The construction was then loaded until failure under a displacement rate of 1 mm per second. RESULTS: There were no significant differences concerning the maximum pullout force (group I: 357 N +/- 61; group II: 326 N +/- 92; group III: 343 N +/ 55). In case of the expansion bolt, we found the stiffness to be higher (61 N/mm) when compared to group I (48 N/mm), and group II (52N/mm) (P < 0.01 I vs. III). Using interference screws, we were able to demonstrate a strong correlation between torque and pullout forces (group I: r2 = 0.7; group II: r2 = 0.92). Ruptures of the suturing material occurred only in groups I and II. CONCLUSION: We conclude that hamstring graft fixation, using the presented expansion bolt, demonstrates fixation strength similar to the established screw fixation and can therefore be regarded as a reasonable alternative fixation method. Especially, since some specific disadvantages of screw fixation can be prevented by application of the bolt fixation. PMID- 16044295 TI - Quantification of aortic elasticity: development and experimental validation of a method using computed tomography. AB - Aortic distensibility depending on aortic cross-sectional area changes is an important parameter for the grading of vascular diseases. This study measured aortic area changes by multidetector computed tomography. An image reconstruction algorithm was developed to assess aorta diameter and area as a function of the cardiac cycle with sufficient time resolution along the entire length of the aorta by four-detector row computed tomography. The algorithm was tested on porcine aortic specimens and compared with an optical reference method. The error of the relative vessel area change comparing the two methods was found to be about 3%. Initial tests on patient datasets indicate that clinical application is feasible. The proposed method has the advantage that it can easily be integrated into a modified routine CT angiography study and allows the measurement of aortic cross-sectional area changes. PMID- 16044296 TI - Numbers and space: a cognitive illusion? AB - This study investigated the relationships between numerical and spatial representations by means of a bisection task, exploring the view that the core representation of number meaning is spatially organized as a mental number line. In Experiment nos. 1 (bisection of digit strings) and 2 (bisection of flanked lines) spatial biases towards the larger digit were found to be related only to processing of relative magnitude. Experiment nos. 3 (bisection of an unfilled space) and 4 (bisection of flanked lines/unfilled spaces) aimed at disclosing perceptual, attentional, and numerical constraints on the bias induced by the position of the larger digit. This effect is interpreted in terms of a cognitive illusion of length, whereby a spatial bias compensates for the numerical disparity. This seems to operate in a categorical fashion ("small/large"), and to be congruent with the assumption that relatively large numbers are associated with the right side of a mental representational space. PMID- 16044297 TI - Sustained attention in traumatic brain injury (TBI) and healthy controls: enhanced sensitivity with dual-task load. AB - Poor sustained attention or alertness is a common consequence of traumatic brain injury (TBI) and has a considerable impact on the recovery and adjustment of TBI patients. Here, we describe the development of a sensitive laboratory task in healthy subjects (Experiment 1) and its enhanced sensitivity to sustained attention errors in TBI patients (Experiment 2). The task involves withholding a key press to an infrequent no-go target embedded within a predictable sequence of numbers (primary goal) and detecting grey-coloured targets within the sequence (secondary goal). In Experiment 1, we report that neurologically healthy subjects are more likely to experience a lapse of attention and neglect the primary task goal, despite ceiling performance on the secondary task. Further, attentional lapses on the task correlated with everyday attentional failures and variability of response time. In Experiment 2, the task discriminates between TBI patients and controls with a large effect size. The dual-task yields more errors in both groups than a simple task involving only the primary goal that is commonly used to detect sustained attention deficits in neurologically impaired groups. TBI patients' errors also correlated with everyday cognitive failures and variability of response time. This was not the case in the simple version of the task. We conclude that the dual-task demand associated with this task enhances its sensitivity as a measure of sustained attention in TBI patients and neurologically healthy controls that relates to everyday slips of attention. PMID- 16044298 TI - Corticospinal facilitation during first and third person imagery. AB - Motor imagery can be defined as the covert rehearsal of movement. Previous research with transcranial magnetic stimulation (TMS) has demonstrated that motor imagery increases the corticospinal excitability of the primary motor cortex in the area corresponding to the representation of the muscle involved in the imagined movement. This research, however, has been limited to imagery of oneself in motion. We extend the TMS research by contrasting first person imagery and third person imagery of index finger abduction-adduction movements. Motor evoked potentials were recorded from first dorsal interosseous (FDI) and abductor digiti minimi (ADM) during single pulse TMS. Participants performed first and third person motor imagery, visual imagery, and static imagery. Visual imagery involved non biological motion while static imagery involved a first person perspective of the unmoving hand. Relative to static imagery, excitability during imagined movement increased in FDI but not ADM. The facilitation in first person imagery adds to previous findings. A greater facilitation of MEPs recorded from FDI was found in third person imagery where the action was clearly attributable to another person. We interpret this novel result in the context of observed action and imagined observation of self action, and attribute the result to activation of mirror systems for matching the imagined action with an inner visuo-motor template. PMID- 16044299 TI - Projections of diencephalic dopamine neurons into the spinal cord in mice. AB - The aim of this study is to investigate the pathway of diencephalic dopaminergic (DA) neuronal innervating into the spinal cord in mice, the pathway is postulated relevant to clinical restless legs syndrome (RLS). Tyrosine hydroxylase (TH) immunohistochemistry was used to identify the DA neuron. The fluorescent tracer Fluoro-Gold (FG) was stereotaxically injected into the T10-L5 spinal cord of CBL57 mice (n=20) seven days before the animals were sacrificed. The diencephalic sections were stained with TH antibody and the FG tracer present in the diencephalic DA neurons were examined under fluoresce microscope. The average number of total DA neurons per side in A11, A12, A13 and A14 was 66+/-8, 221+/ 12, 350+/-17 and 254+/-21 respectively. After being injected into the spinal cord, FG reached the DA neurons within the A10 and A11 groups, but didn't target to any other DA neuron groups including the A8 and A9 groups in substantia nigra (SN). The diencephalic A11 DA neurons possessed long axons extending over several segments and possibly traversing the entire length of the spinal cord. It is the first time to report A10 and A11 DA neuron projections into the spinal cord in mice. PMID- 16044300 TI - The effect of clonidine on cell survival, glutamate, and aspartate release in normo- and hyperglycemic rats after near complete forebrain ischemia. AB - The present study was undertaken to investigate the effects of the alpha2 adrenergic agonist, clonidine, on the near complete cerebral ischemia (NCFI) evoked release of glutamate and aspartate from normo- and hyperglycemic rodent brain tissue using microdialysis tissue techniques. Hemodynamic variables, blood lactate, and glucose levels were monitored throughout the 40 min NCFI occlusion period. After 48 h, rats were killed and the extent of neuronal injury was determined in the cortex, striatum, and hippocampus. Hemodynamic variables recorded during ischemia improved with clonidine treatment in both normo- and hyperglycemic groups. Glutamate and aspartate levels were greatly increased over control values during normo- and hyperglycemic NCFI treatment. Clonidine pretreatment suppressed the release of both glutamate and aspartate during NCFI in normo- and hyperglycemic rodents when compared with NCFI-treated normo- and hyperglycemic rats without the drug. Significant neuroprotection of cells in the cortex, striatum, and hippocampus was also observed in drug-treated animals 48 h postischemia. The combined effects of diminished glutamate release after NCFI and reduced neuronal injury in both normo- and hyperglycemic states suggests that clonidine treatment during NCFI is neuroprotective. The neuroprotective effect of clonidine during ischemia may be ascribed to both a sensitization of central sympathetic activity and a reduced release of glutamate thereby reducing NMDA receptor activation and neuronal damage. PMID- 16044301 TI - Is proprioception calibrated during visually guided movements? AB - In the present study, a visuomotor adaptation paradigm was used to determine whether proprioception is calibrated during visually guided movements. A full vision group (FV) was given vision of a cursor representing hand position and was asked to aim towards visible targets. A no vision group (NV) performed the same task without vision of the cursor and was given knowledge of results (KR) after movement completion. A directional bias was introduced between the location of the cursor and the location of the hand in an adaptation phase, which resulted in a deviation to the right of the intended target. Of interest was whether participants would still show rightward deviations in a post-test series of trials in which vision was removed (aftereffects). The NV group presented strong aftereffects. However, the FV group only showed modest aftereffects early in the post-test, which rapidly decayed over the course of the post-test. Further analyses showed that the presence of those early aftereffects was due to an offline influence of vision on movement planning. PMID- 16044302 TI - Change perception using visual transients: object substitution and deletion. AB - In three experiments we studied change detection and identification when no extraneous transients were present in the image at the time of change. Each image consisted of 12 different objects, sorted by color into three different levels of probability of change. In Experiment 1, change of one object was detected and identified frequently in objects having the highest probability of change (central interest), which we hypothesize were mainly visited by attention. Changes in other objects with a lower probability of change (marginal interest), however, although detected efficiently were unlikely to be identified. Identification improved for less attended objects if the changed stimulus simply disappeared, allowing visual persistence to hold information about the object until attention could be shifted to it (Experiment 2). Contrary to previous findings showing that response times (RTs) for luminance change detection in a multi-element display are not altered by attention, we found changes in objects of central interest to be detected faster than in objects of marginal interest when objects' identity was to be held in working memory. However, no differences in RTs emerged in the same change detection task when objects' identity was not stored in working memory (Experiment 3). PMID- 16044303 TI - Impaired balancing ability in dyslexic children. AB - Children with developmental dyslexia struggle to learn to read and spell despite adequate intelligence and educational opportunity. Several lines of research are attempting to establish the neurobiological basis of dyslexia, and low-level sensory and motor deficits have been found in dyslexic populations; furthermore, behavioural and imaging data point to cerebellar dysfunction in dyslexia. To investigate this, normal readers (n=19) and children with developmental dyslexia (n=16) were asked to perform various cognitive, literacy, and balancing tasks. Children balanced on the left or right foot, with eyes open or closed, for a period of 10 s during which their movements were recorded with a motion-tracking system. Dyslexic children were less stable than the control children in both eyes open conditions (left foot P=0.02, right foot P=0.012). While there were no group differences during the eyes-closed conditions, the dyslexic children dropped a foot to correct balance significantly more often than control children (P<0.05). Incidence analysis showed that 50% of the dyslexic group fell into the 'impaired' category on the eyes-open balancing tasks; when the mean balancing scores and the foot drops were considered, only three of our dyslexic children showed no evidence of balancing difficulties. There were strong correlations between reading and spelling scores and the mean eyes-open balancing score (r=0.52 and 0.44, respectively). Thus, while not all children with developmental dyslexia show impaired balancing skills, low-level motor dysfunction may be associated with impaired literacy development. This could be due to several factors, including the involvement of the cerebellum, the magnocellular system, or more general developmental immaturity. PMID- 16044304 TI - Target selection in eye-hand coordination: Do we reach to where we look or do we look to where we reach? AB - During a goal-directed movement of the hand to a visual target the controlling nervous system depends on information provided by the visual system. This suggests that a coupling between these two systems is crucial. In a choice condition with two or more equivalent objects present at the same time the question arises whether we (a) reach for the object we have selected to look at or (b) look to the object we have selected to grasp. Therefore, we examined the preference of human subjects selecting the left or the right target and its correlation to the action to be performed (eye-, arm- or coordinated eye-arm movement) as well as the horizontal position of the target. Two targets were presented at the same distance to the left and right of a fixation point and the stimulus onset asynchrony (SOA) was adjusted until both targets were selected equally often. This balanced SOA was then taken as a quantitative measure of selection preference. We compared these preferences at three horizontal positions for the different movement types (eye, arm, both). The preferences of the 'arm' and 'coordinated eye-arm' movement types were correlated more strongly than the preferences of the other movement types. Thus, we look to where we have already selected to grasp. These findings provide evidence that in a coordinated movement of eyes and arm the control of gaze is a means to an end, namely a tool to conduct the arm movement properly. PMID- 16044305 TI - Magnetoencephalographic signals predict movement trajectory in space. AB - Brain-machine interface (BMI) efforts have been focused on using either invasive implanted electrodes or training-extensive conscious manipulation of brain rhythms to control prosthetic devices. Here we demonstrate an excellent prediction of movement trajectory by real-time magnetoencephalography (MEG). Ten human subjects copied a pentagon for 45 s using an X-Y joystick while MEG signals were being recorded from 248 sensors. A linear summation of weighted contributions of the MEG signals yielded a predicted movement trajectory of high congruence to the actual trajectory (median correlation coefficient: r=0.91 and 0.97 for unsmoothed and smoothed predictions, respectively). This congruence was robust since it remained high in cross-validation analyses (based on the first half of data to predict the second half; median correlation coefficient: r=0.76 and 0.85 for unsmoothed and smoothed predictions, respectively). PMID- 16044306 TI - Muscle activation and time to task failure differ with load type and contraction intensity for a human hand muscle. AB - Time to failure for sustained isometric contractions of the elbow flexors is briefer when maintaining a constant elbow angle while supporting an inertial load (position task) compared with exerting an equivalent torque against a rigid restraint (force task). Our primary purpose was to determine whether the effects of load type on time to task failure exist when motor unit recruitment cannot be enhanced during a sustained submaximal contraction of an intrinsic hand muscle. A second purpose was to determine whether a greater reserve remains in the muscle after early failure of the position task. Two groups of 10 strength-matched men performed the force and position tasks at either 20% or 60% of maximal force (MVC) with the first dorsal interosseus, followed by a second force task at the same relative intensity. The rate of increase in surface EMG was greater (P = 0.002) and time to failure was briefer (P = 0.005) for the position task (593 +/- 212 s) compared with the force task (983 +/- 328 s) at 20% MVC, whereas there were no task differences in these variables at 60% MVC (P >or= 0.200). Time to failure for the second force tasks did not differ at either contraction intensity (P>or=0.743). These results demonstrate that previously observed effects of load type generalize to a hand muscle, although only for low-intensity contractions. For the position task at low forces, muscle activity increased more rapidly and no additional reserve remained in the muscle at failure compared with the force task. We propose that the briefer time to failure for the position task during sustained, low-intensity contractions is due to earlier recruitment of the motor unit pool. PMID- 16044308 TI - TRPMs and neuronal cell death. AB - Death of CNS neurons during acute injury occurs as a result of a complex combination of excitotoxicity, necrosis, apoptosis, oedema and inflammatory reactions. Neuroprotection via glutamate receptor blockade or antioxidant or anti inflammatory therapy have not proven effective in the clinical treatment of brain damage due to narrow therapeutic windows, poor pharmacokinetics or blockade of the signalling essential for normal excitatory neurotransmission and neuronal survival. Recent work in neuronal biochemistry, genomics and proteomics has increased understanding of the molecular organization of the excitatory synapse and the neuronal postsynaptic density. Transient receptor potential (TRP) channels are an exciting new family of cation channels that are highly expressed in the brain. Several members can be induced by oxidative stress and oxygen free radicals, both of which play important roles in neurodegeneration. Recent work has indicated that members of the melastatin subfamily (TRPM) of TRP proteins, particularly TRPM7 and TRPM2, may play key roles in neuronal death that is activated by oxidative stress and downstream from excitotoxic signal pathways. This discovery provides an exiting new avenue for research into the pathophysiology and treatment of acute neurodegeneration. PMID- 16044307 TI - Protein-protein interaction and functionTRPC channels. AB - Since their identification in the concluding years of the last century, the mammalian transient receptor potential (canonical) (TRPC) channels have remained in the limelight as the primary candidates for the Ca(2+) entry pathway activated by the hormones, growth factors, and neurotransmitters that exert their effect through activation of PLC. Although TRPC channels have been shown clearly to mediate, at least in part, receptor-activated Ca(2+) entry in literally all cell types, several of their central characteristics, as recorded in expression systems using recombinant channels, differ from those of the native receptor dependent Ca(2+) influx channels. The present review attempts to highlight the interaction of TRPC channels with other proteins, which may explain the variability of TRPC channel activation and regulatory mechanisms observed with the native and recombinant channels. These include the homologous and heterotopous interactions of TRPC channel isoforms, the interaction of TRPC channels with calmodulin, PLCgamma, IP(3) receptors, and with scaffolding proteins like InaD, EBP50/NEHRF, caveolin, Janctate and Homers. PMID- 16044310 TI - Distribution of peptidase activity in teleost and rat tissues. AB - Peptides play important roles in cell regulation and signaling in many tissues. The actions of peptides are regulated by peptidases. Although the activity of these enzymes has been thoroughly characterized in mammals, little is known about their presence or function in fish. In the present study, we compared the activity of several peptidases in selected tissues (pituitary gland, different brain areas, kidney and gills) of the gilthead sea bream and rainbow trout with that found in similar rat tissues (lungs studied in place of gills). Soluble puromycin-sensitive aminopeptidase showed the highest values in the pituitary gland of the sea bream, whereas the membrane-bound form was found to be more active in the trout kidney. Very high levels of activity of aminopeptidase N were detected in trout and sea bream plasma. In contrast, the highest levels of activity of aminopeptidase B were found in rat tissues, with the exception of the gills of the trout. Aminopeptidase N levels tended to be higher in sea bream tissues with respect to those of trout. In contrast, the level of activity of aminopeptidase B was found to be consistently much higher in trout tissues than in those of the sea bream. Prolyl endopeptidase activity was principally detected in the pituitary gland and in the brain areas of teleosts. These differences between species could be related to different mechanisms of osmoregulation in saltwater- and in freshwater-adapted fish. PMID- 16044309 TI - TRPV5 and TRPV6 in Ca(2+) (re)absorption: regulating Ca(2+) entry at the gate. AB - Many physiological functions rely on the exact maintenance of body Ca(2+) balance. Therefore, the extracellular Ca(2+) concentration is tightly regulated by the concerted actions of intestinal Ca(2+) absorption, exchange of Ca(2+) to and from bone, and renal Ca(2+) reabsorption. Renal distal convoluted and connecting tubular cells as well as duodenal epithelial cells are unique in their ability to mediate transcellular (re)absorption of Ca(2+) at large and highly variable rates. Two members of the transient receptor potential (TRP) superfamily, TRP vanilloid (TRPV)5 and TRPV6, are specialized epithelial Ca(2+) channels responsible for the critical Ca(2+) entry step in transcellular Ca(2+) (re)absorption in intestine and kidney, respectively. Because transcellular Ca(2+) transport is fine-tuned to the body's specific requirements, regulation of the transmembrane Ca(2+) flux through TRPV5/6 is of particular importance and has, therefore, to be conspicuously controlled. We present an overview of the current knowledge and recent advances concerning the coordinated regulation of Ca(2+) influx through the epithelial Ca(2+) channels TRPV5 and TRPV6 in transcellular Ca(2+) (re)absorption. PMID- 16044311 TI - Secondary Ph+ acute lymphoblastic leukemia after temozolomide. PMID- 16044312 TI - Hyperbilirubinemia and cholelithiasis in Chinese patients with hemoglobin H disease. AB - Hemoglobin H disease (HbH) is a hemoglobinopathy peculiar to parts of the world with high incidence alpha-thalassemia mutations. Among 90 HbH cases, 50 cases suffered from clinically significant jaundice (bilirubin >30 mmol/l), including 14 with severe jaundice (bilirubin >60 mmol/l). Cholelithiasis was found in 38 cases. The incidence is roughly eight times higher than that in background control population but 50% lower than that in beta-thalassemia. The risk of gallstones was related to higher bilirubin levels but not alpha-globin genotype, sex, ferritin, and hemoglobin levels. Homozygotes or double heterozygotes for Gilbert alleles (17.2%), but not heterozgyotes (42.2%), were found to have a significantly increased risk of gallstones and jaundice. However, common Chinese Gilbert syndrome alleles do not completely explain the variable risks. PMID- 16044313 TI - AML bearing the translocation t(11;17)(q23;q21): involvement of MLL and a region close to RARA, with no differentiation response to retinoic acid. AB - We describe a case of acute myeloid leukemia (AML) bearing the translocation t(11;17)(q23;q21). The morphological phenotype represented a monoblastic leukemia, AML French-American-British (FAB) M5a. Further analysis of the translocation revealed an involvement of the mixed-lineage leukemia (MLL) gene and a region closely proximal to the retinoic acid (RA) receptor alpha (RARA) gene. AMLs involving both a rearranged MLL and the 17q21 region, in which the RARA gene is located, have only been described in some individual cases. The functional role of this translocation is still unknown. Rearrangements of the MLL (11q23) gene in AML are usually related to the morphological phenotype FAB M5. In general, they are associated with an adverse prognosis. In acute promyelocytic leukemia, the translocation (15;17)(q22;q11-21) involving the RARA leads to a maturation arrest that can be overcome by RA, often inducing remission. In other forms of AML, however, the effects of RA are limited and diverse. To study whether RA might have a therapeutical potential in our case, we performed an in vitro analysis of RA effects on AML cells. We found that RA leads to enhanced cell death and up-regulation of CD38 and CD117. However, no hints of RA-induced in vitro differentiation were visible. Our data indicate that in AML cells bearing the t(11;17)(q23;q21), a differentiation arrest that is overcome by RA is not present. On the contrary, RA induces alterations in cellular regulation that are similar to the RA-induced changes observed in early hematogenic progenitors; thus, a possible therapeutical benefit of RA in such cases remains open. PMID- 16044314 TI - A case of deep vein thrombosis in patient with Hermansky-Pudlak syndrome. PMID- 16044315 TI - Use of recombinant factor VIIa in the management of severe bleeding episodes in patients with Bernard-Soulier syndrome. AB - Bernard-Soulier syndrome (BSS) is a rare congenital platelet disorder characterized by defective platelet adhesion and manifested by spontaneous and often profuse bleeding. Recombinant factor VIIa (rFVIIa) is a haemostatic agent licensed for the treatment of bleeding episodes in patients with haemophilia and inhibitors, which may represent a low-risk alternative to existing therapies in the management of patients with BSS. Here, we describe the use of rFVIIa for the treatment of three severe bleeding episodes in two patients with BSS. Data were extracted by automated searching of the international, Internet-based registry http://www.haemostasis.com . Patient 1, a 24-year-old woman, was admitted with severe epistaxis and hypotension. The diagnosis of BSS was confirmed by macrothrombocytopenia, absence of ristocetin-induced platelet agglutination (RIPA) and absence of glycoprotein (GP) Ibalpha and IX on the platelet surface. Epsilon aminocaproic acid (EACA; two 50-mg/kg doses), packed red blood cells (PRBCs, 2 U) and platelets (30 U) failed to control the bleeding and, after 13 h, three bolus doses of rFVIIa (90 microg/kg body weight) and a third dose of EACA were administered; bleeding stopped after the third dose of rFVIIa. Patient 2, a 15-year-old girl, initially presented with severe menorrhagia. A lack of RIPA and severe deficiency of GPIbalpha on the platelet surface confirmed the diagnosis of BSS. EACA and fresh-frozen plasma did not control the haemorrhage, but two bolus doses of rFVIIa (98 microg/kg body weight) resulted in a marked decrease in bleeding. On second admission, patient 2 had severe epistaxis and mild menorrhagia. Two rFVIIa doses (98 and 122.5 microg/kg body weight) were given, and the bleeding stopped. No adverse events were reported in these cases. These three admissions highlight the potential of rFVIIa for the treatment of severe bleeds in patients with BSS. PMID- 16044316 TI - The influence of the response-stimulus interval on implicit and explicit learning of stimulus sequence. AB - Three experiments investigated the influence of the response-stimulus interval (RSI) on implicit and explicit learning of stimulus sequences. Participants responded to numerals presented in predetermined positions with alternating long and short RSIs. Half of the participants were instructed explicitly to learn the position sequence. In the transfer phase of Experiments 1 and 2, changing RSI patterns reduced the expression of incidental and intentional learning of position sequence. In Experiment 3 the position sequence was transformed, except that sub-sequences demarcated by long RSIs remained unchanged; this greatly reduced the expression of intentional learning, and slightly reduced that of incidental learning. These results indicate that in implicit learning, stimulus sequences are learned under the constraints of RSIs, whereas in explicit learning, learning independent of RSIs, as well as learning constrained by RSIs, occurs. PMID- 16044317 TI - Ultrastructural evidence of piecemeal degranulation in large dense-core vesicles of brain neurons. AB - Large dense-core vesicles (LDCV) are a group of neuronal secretory organelles with different size and characteristically condensed morphology. LDCV release their specific cargo by regulated exocytosis, either in the form of "full fusion" or "kiss-and-run" exocytosis. In this paper, we provide ultrastructural evidence indicative of a slow and particulate mode of secretion from LDCV, called piecemeal degranulation (PMD). A number of LDCV in the nerve boutons of mouse brain presented marked increase in their size accompanied by reduction and also disappearance of content material. Residual secretory constituents in altered LDCV displayed eroded marginated patterns, leading to eccentric "haloed" morphologies. Remarkably, altered LDCV never appeared to be fused with each other or with the nerve plasma membrane. Very small vesicles, empty or apparently loaded with the same material making-up the LDCV content, could be seen near or attached to LDCV and the plasma membrane. First described in basophils, mast cells and eosinophils, PMD has recently been recognized in various neuro endocrine cells, like adrenal chromaffin cells and endocrine cells of the gastro intestinal epithelia. Here we suggest that PMD may be a hitherto unrecognized pathway of neuron secretion. It would represent the morphological correlate of a long-lasting and low-level process of neuro-transmitter release. It extends the patterns of neuron secretion and possibly opens new perspectives in understanding neuron plasticity. PMID- 16044318 TI - Embryonic development of choline acetyltransferase and nitric oxide synthase in the spinal cord of pigeons and chickens with special reference to the superficial dorsal horn. AB - The superficial dorsal horn of birds as well as mammals contains both cholinergic and nitrergic neuronal structures as evident from the presence of the synthesizing enzymes such as choline acetyltransferase and nitric oxide synthase, which is an NADPH diaphorase. In the rat, both systems develop only postnatally. Rats are altricial at birth whereas pigeons and chickens are semiprecocial or precocial, respectively, at the time of hatching. Immunocytochemical studies of choline acetyltransferase and nitric oxide synthase in the developing avian spinal cord (starting with embryonic day 12 of 18 in the pigeon and 14 of 21 in the chicken) showed that both systems are well developed in the superficial dorsal horn at the time of hatching in both avian species. In the pigeon, choline acetyltransferase-positive superficial dorsal horn neurons appear only on the day of hatching (E18), whereas nitric oxide synthase-positive neurons can be first detected at stage E14. In the chicken, nitric oxide synthase-positive neurons are present already at stage E14, whereas choline acetyltransferase-positive neurons appear at stage E20. Autonomic and somatic motor neurons show adult-like choline acetyltransferase-immunoreactivity and/or nitric oxide synthase-immunoreactivity at the earliest stages investigated. It is concluded that the stage of maturation at birth or hatching plays an important role in the development of superficial dorsal horn cholinergic and nitrergic systems. PMID- 16044319 TI - Structural and mechanical architecture of the intestinal villi and crypts in the rat intestine: integrative reevaluation from ultrastructural analysis. AB - The ultrastructure of the rat intestinal interstitium was analyzed from the viewpoint of mechanical dynamics to stabilize the intestinal villi, crypts and mucosal folds. In the rat, the small intestine lacks circular folds, but the large intestine possesses spiral folds. The intestinal villi, the largest in the duodenum, decreased in size in the jejunum and ileum successively, and were absent in the large intestine. The intestinal interstitium consisted of lamina propria mucosae (LPM) and tela submucosa (TSM) separated by muscularis mucosae (MM), the LPM was subdivided into an upper part within the villi and a lower part among the crypts in the small intestine. The light microscopic density of interstitium in the intestinal wall was lowest in the upper LPM, moderately dense in the lower LPM and highest in the TSM, and that among the intestinal region was highest in the duodenum and decreased successively in the jejunum and ileum. In the large intestine, the TSM bulged to form spiral folds with very low density. The intestinal epithelium in the villi possessed wide intercellular spaces and that in the crypts had closed intercellular spaces. At electron microscopic level, the upper and lower LPM contained subepithelial supportive meshwork that consisted of collagen fibrils and myofibroblast processes. The lower LPM and TSM contained conspicuous bundles of collagen fibrils and, in addition, TSM contained minor populations of scattered collagen fibrils near the smooth muscle layer (SML). The diameter of collagen fibrils was the largest in the bundles of TSM, and decreased from the duodenum through the jejunum and ileum to the large intestine. On the basis of these observations, we hypothesize that the intestinal villi are mechanically stabilized by the balance between the expansive interstitial pressure and inward pull by the subepithelial supportive meshwork. This hypothesis explains the hitherto neglected fact that the intestinal epithelium possesses wide intercellular spaces only in the villi, and accounts for the counterforce against the perpendicular smooth muscle cells, which are supposed to contract the intestinal villi. PMID- 16044320 TI - Distribution of TRPC1 and TRPC5 in medial temporal lobe structures of mice. AB - The transient receptor potential (TRP) superfamily comprises a group of non selective cation channels that have been implicated in both receptor and store operated channel functions. The family of the classical TRPs (TRPCs) consists of seven members (TRPC1-7). The presence of TRPC1 and TRPC5 mRNA in the brain has previously been demonstrated by real-time polymerase chain reaction. However, the distribution of these receptors within different brain areas of mice has not been investigated in detail. We have used antibodies directed against TRPC1 and TRPC5 to study the distribution and localization of these channels in murine medial temporal lobe structures. Both TRPC1 and TRPC5 channels are present in the various nuclei of the amygdala, in the hippocampus, and in the subiculum and the entorhinal cortex. We have found that TRPC1 channels are primarily expressed on cell somata and on dendrites, whereas TRPC5 channels are exclusively located on cell bodies. Moreover, TRPC1 channels are selectively expressed by neurons, whereas TRPC5 channels are mainly expressed by neurons, but also by non-neuronal cells. The expression of TRPC1 and TRPC5 channels in mammalian temporal lobe structures suggests their involvement in neuronal plasticity, learning and memory. PMID- 16044321 TI - Free fatty acid receptor 1 (FFA(1)R/GPR40) and its involvement in fatty-acid stimulated insulin secretion. AB - Free fatty acids (FFA) have generally been proposed to regulate pancreatic insulin release by an intracellular mechanism involving inhibition of CPT-1. The recently de-orphanized G-protein coupled receptor, FFA(1)R/GPR40, has been shown to be essential for fatty-acid-stimulated insulin release in MIN6 mouse insulinoma cells. The CPT-1 inhibitor, 2-bromo palmitate (2BrP), was investigated for its ability to interact with mouse FFA(1)R/GPR40. It was found to inhibit phosphatidyl inositol hydrolysis induced by linoleic acid (LA) (100 muM in all experiments) in HEK293 cells transfected with FFA(1)R/GPR40 and in the MIN6 subclone, MIN6c4. 2BrP also inhibited LA-stimulated insulin release from mouse pancreatic islets. Mouse islets were subjected to antisense intervention by treatment with a FFA(1)R/GPR40-specific morpholino oligonucleotide for 48 h. Antisense treatment of islets suppressed LA-stimulated insulin release by 50% and by almost 100% when islets were pretreated with LA for 30 min before applying the antisense. Antisense treatment had no effect on tolbutamide-stimulated insulin release. Confocal microscopy using an FFA(1)R/GPR40-specific antibody revealed receptor expression largely localized to the plasma membrane of insulin-producing cells. Pretreating the islets with LA for 30 min followed by antisense oligonucleotide treatment for 48 h reduced the FFA(1)R/GPR40 immunoreactivity to background levels. The results demonstrate that FFA(1)R/GPR40 is inhibited by the CPT-1 inhibitor, 2BrP, and confirm that FFA(1)R/GPR40 is indeed necessary, at least in part, for fatty-acid-stimulated insulin release. PMID- 16044322 TI - Severe vitreous hemorrhage associated with closed-globe injury. AB - BACKGROUND: The presence of marked vitreous hemorrhage is associated with poor visual prognosis in ocular trauma involving the posterior segment. The purpose of this study was to investigate the clinical features, prognostic factors, and visual outcomes in such eyes after closed-globe injury. METHODS: Retrospective chart review of patients with severe vitreous hemorrhage after closed-globe injury between January 2000 and December 2002. RESULTS: The study included 33 eyes in 32 patients with a mean age of 41.5 (range, 3-78 years) and a median follow-up period of 12 (range, 3-50 months). Presenting visual acuity was between 4/200, and there was light perception in 32 eyes (97%) and no light perception in one eye (3%). Fifteen eyes (45%) had one or more posterior segment pathologies beneath the vitreous hemorrhage. There were retinal tears in six eyes (18%), retinal detachment in six (18%), submacular hemorrhage in six (15%), choroidal rupture in three (9%), macular pucker in three (9%), retinal dialysis in one (3%), macular hole in one (3%), and retinal vessels occlusion in one (3%). Seven eyes (21%) had final visual acuity 20/40 or better, and 18 eyes (54%) had less than 20/200. Poor prognostic factors included presenting visual acuity of light perception or lower (p=0.032), hyphema (p=0.007), traumatic cataract (p=0.027), and age 55 years or younger (p=0.039). The most common cause of poor visual outcome was macular scar. CONCLUSION: The visual prognosis of severe vitreous hemorrhage after closed-globe injury was guarded. This was frequently associated with various anterior and posterior segment pathologies. In order to provide early visual rehabilitation and management of any potentially treatable posterior pathology, early vitrectomy may be considered. PMID- 16044323 TI - Bactericidal/permeability-increasing protein in lacrimal gland and in tears of healthy subjects. AB - BACKGROUND: To determine the expression of bactericidal/permeability-increasing protein (BPI), a novel antimicrobial molecule, in the main lacrimal gland and its content in tears of young healthy subjects. METHODS: BPI concentration of tears was measured in 42 healthy volunteers, 13 men and 29 women, with ages ranging from 22 to 30 (mean 24.7+/-2.1) years by a time-resolved fluoroimmunoassay (TR FIA). Immunohistochemical analysis was made to localize BPI in lacrimal gland and conjunctiva of eight autopsied subjects, two men and six women, with the age range from 44 to 87 (mean 72.3+/-14.9) years. RESULT: The mean concentration of BPI in tears was 27.8+/-29.5 microg/l, and it decreased with an increase in tear flow rate (P<0.0001). There was no statistically significant difference in BPI content of tears between the genders. BPI was immunohistochemically seen in outer basal epithelial cells of intralobular and excretory ducts, squamous and basal cells of conjunctiva as well as faintly in myoepithelial cell layer of acini. The presence of BPI in the lacrimal gland and in the tear fluid was verified by Western blotting. CONCLUSIONS: The results indicate that outer basal epithelial cells of lacrimal gland ducts contain BPI, which occurs in a relatively high concentration in tears. BPI may have a substantial antibacterial role in human tears. PMID- 16044324 TI - A rare case of choroidal neovascularization following macular hole surgery. AB - BACKGROUND: Choroidal neovascularization occurs in a wide spectrum of conditions, including degenerative, inflammatory, traumatic and hereditary conditions, all of which are characterized by breaks in Bruch's membrane. A few cases of choroidal neovascular membrane (CNVM) have been reported in the literature following macular hole surgery. METHOD: We present a retrospective case study of a single patient who developed CNVM after successful macular hole surgery. The 70-year-old diabetic female underwent macular hole surgery in her right eye. Posterior hyaloid was separated with suction and vitrectomy was performed. Internal limiting membrane (ILM) was stained with trypan blue under air and ILM peeling was performed. Perfluoropropane (C3F8) gas was used as the tamponade. Six weeks post-operation, closure of macular hole was seen. Six months later, she presented with defective vision. Subfoveal classic CNVM was seen in the same eye. DISCUSSION: About 1%-3% of patients who undergo macular hole surgery develop CNVM. Age-related changes and surgical trauma are considered to be the predisposing factors in reported cases. Injury to the retinal pigment epithelium (RPE) during surgery may be an important factor in our case, since there were no pre-existing age-related changes. CONCLUSIONS: CNVM though rare can occur after macular hole surgery, especially in the setting of age-related changes. Injuries to the RPE should be avoided during surgery. We report a case of CNVM after macular hole in which trypan blue was used as a staining agent. PMID- 16044325 TI - Inhibitory effect of triamcinolone acetonide on corneal neovascularization. AB - BACKGROUND: Corneal neovascularization (NV) plays an important role in the pathogenesis of corneal disorders. Recently, triamcinolone acetonide (TA) has been reported as a potential treatment for ocular angiogenesis. However, there are no reports on the inhibitory effect of TA on the corneal NV. METHODS: Triamcinolone acetonide (2 mg) was administered to four rabbits' eyes by a subconjunctival injection immediately after a basic fibroblast growth factor (bFGF)-pellet was placed into the cornea. As a control, four eyes received an injection of distilled water. Four weeks later, the inhibition of corneal NV was evaluated as the percentage ratio of the vessel invasion area to the area that was sandwiched between the pellet and the limbus cornea. To identify the characteristic appearance of new corneal vessels, the control cornea was examined by using the antibody of vascular endothelial growth factor (VEGF). To confirm TA concentration in TA-treated corneas, the TA level was measured using high performance liquid chromatography. RESULTS: Neovascularization from the limbus to the pellet was detected in control eyes 4 weeks after the bFGF pellet implantation. TA-treated eyes demonstrated the inhibition of the neovascular response to the pellet. The severity of NV as compared between control and TA treated eyes was statistically significant (P<0.05). Morphologically, new vessel growth was shown in the control cornea, and endothelial cells of new vessels were positively stained with the antibody of VEGF. TA concentration in TA-treated corneas at 2 weeks showed 63.5+/-42.8 microg/g (n=4, mean +/- SD), while TA was not detected in control and TA-treated corneas at 4 weeks. The level of TA was effectively maintained for at least 2 weeks after the subconjunctival injection. CONCLUSION: We have demonstrated that subconjunctival TA administration inhibited rabbit corneal NV. This agent may prove useful in the treatment of corneal angiogenic disorders. PMID- 16044326 TI - Comparison of confocal scanning laser ophthalmoscopy, scanning laser polarimetry and optical coherence tomography to discriminate ocular hypertension and glaucoma at an early stage. AB - BACKGROUND: The aim was to compare the ability of confocal scanning laser ophthalmoscopy (CSLO), scanning laser polarimetry (SLP), and optical coherence tomography (OCT) to discriminate eyes with ocular hypertension (OHT), glaucoma suspect eyes (GS) or early glaucomatous eyes (EG) from normal eyes. METHODS: Ocular hypertension, GS, and EG were defined as normal disc with intraocular pressure >21 mmHg, glaucomatous disc without visual field loss, and glaucomatous disc accompanying the early glaucomatous visual filed loss respectively. Ninety three normal eyes, 26 OHT, 55 GS, and 67 EG were enrolled. Optic disc configuration was analyzed by CSLO (version 3.04), whereas retinal nerve fiber layer thickness was analyzed by SLP (GDx-VCC; version 5.3.2) and OCT-1 (version A6X1) in each individual. The measurements were compared in the four groups of patients. Receiver operating characteristic curve (ROC) and area under the curve (AUC) discriminating OHT, GS or EG from normal eyes were compared for the three instruments. RESULTS: Most parameters in GS and EG eyes showed significant differences compared with normal eyes. However, there were few significant differences between normal and OHT eyes. No significant differences were observed in AUCs between SLP and OCT. In EG eyes, the greatest AUC parameter in OCT (inferior--120; 0.932) had a higher AUC than that in CSLO (vertical cup/disc ratio; 0.845; P=0.017). In GS, the greatest AUC parameter in OCT (average retinal nerve fiber layer [RNFL] thickness; 0.869; P=0.002) and SLP (nerve fiber indicator [NFI]; 0.875; P=0.002) had higher AUC than that in CSLO (vertical cup/disc ratio; 0.720). CONCLUSIONS: Three instruments were useful in identifying GS and EG eyes. For glaucomatous eyes with or without early visual field defects, SLP and OCT performed similarly or had better discriminating abilities compared with CSLO. PMID- 16044327 TI - Primary vitrectomy for rhegmatogenous retinal detachment: an analysis of 512 cases. AB - BACKGROUND: Primary pars plana vitrectomy (PPPV) has gained widespread popularity in the treatment of rhegmatogenous retinal detachments (RRD). However, the surgical procedure is still flawed by a significant rate of anatomical and functional failures. The study was conducted to analyse the risk factors for a dissatisfying postoperative outcome. METHODS: We carried out a retrospective study of 512 cases of PPPV with a minimum follow-up of 3 months from one institution over a 9-year period. Preoperatively, 24.8% of patients (127 out of 512) were pseudophakic, 16.4% (84 out of 512) highly myopic, 19.9% (102 out of 512) had preoperative proliferative vitreoretinopathy (PVR) and 14.6% (75 out of 512) had vitreous haemorrhage. RESULTS: The follow-up period ranged from 3 to 108 months (median 14.8). Retinal reattachment was achieved with one operation in 70.7% (362 out of 512) and after one or more operations in 97.5% of cases (499 out of 512). The major reasons for redetachments were new retinal breaks, followed by a combination of new breaks and PVR, and PVR without apparent breaks. Postoperative visual acuities of > or =0.1 and > or =0.4 were achieved in 82.8% (424 out of 512) and 48.2% (247 out of 512) respectively. Out of 376 phakic patients at study entry, 66.4% (250 out of 376) underwent cataract surgery either in combination with PPPV or during the postoperative course. Factors that were significantly associated with either anatomical or functional failure included duration of symptoms, low preoperative visual acuity, myopia, amblyopia, hypotony, macular detachment, preoperative PVR, extent of detachment, involvement of inferior quadrants, no detectable breaks, large breaks, breaks posterior to the equator, surgeon, level of surgical training, endocryotherapy, and combined scleral buckling surgery. CONCLUSION: Primary pars plana vitrectomy is still flawed by a relatively high primary redetachment rate following the initial procedure. The advantages of the technique are a high final reattachment rate and relatively good functional results in a subset of patients with more complicated types of RRD. The risk factors for postoperative failures following PPPV for RRD match to a large extent those following scleral buckling surgery (SBS). Future improvements of the technique will have to focus on modifiable risk factors, such as details of the surgical procedures, surgical training and case selection, to distinguish it from SBS. PMID- 16044328 TI - Incidence of posterior vitreous detachment after laser in situ keratomileusis. AB - BACKGROUND: Vitreoretinal complications are rare in laser in situ keratomileusis (LASIK). Increase in intraocular pressure caused by intraoperative suction with subsequent deforming of the ocular globe and excimer laser shock during the ablation have been discussed as possible causes. The purpose of this study was to determine the effect of LASIK on the vitreous body. PATIENTS AND METHODS: In a prospective study we performed ocular ultrasonography (B scan) immediately before and 1 week after LASIK procedure in 103 myopic or myopic-astigmatic eyes (53 patients, mean age 36.3 years, 32 women, 21 men). In particular, the prevalence, localization, and extent of posterior vitreous detachment (PVD) were determined. RESULTS: The mean spherical equivalent was -4.85 D (range -1.25 to -8.38) and the mean anteroposterior ocular globe length was 25.13 mm (range 23.31-27.65). Ninety five eyes (92.2%) had no PVD preoperatively. Nine eyes out of this group (seven patients, 9.5%) developed incomplete PVD as assessed 1 week postoperatively. Eight eyes (7.8%) had a partial PVD preoperatively and in only one eye was an extension of vitreous detachment observed after the surgery. None of the preoperatively measured parameters could predict the occurrence of PVD by LASIK. CONCLUSIONS: LASIK may in rare cases lead to new occurrence of PVD or extension of a previously existing partial PVD. PMID- 16044329 TI - Specific antibody production in herpes keratitis: intraocular inflammation and corneal neovascularisation as predicting factors. AB - PURPOSE: The purpose of the study is to investigate whether analysis of specific antibody synthesis can aid the diagnosis of herpes keratitis. METHODS: Aqueous humor was collected from 39 patients with presumed recurrent herpes keratitis, including 23 consulting for keratitis and 16 patients scheduled for penetrating keratoplasty. Local antibody production was ascertained by analysis of paired aqueous humor/serum samples, using a modified micro-ELISA technique. RESULTS: Local production of antibodies was found in 32 patients (82%): anti-herpes simplex virus (HSV) antibodies in 26 (67%) and anti-varicella zoster virus (VZV) antibodies in 11 (28%). Twenty of 23 patients with active keratitis (87%), and 12 of 16 undergoing keratoplasty (75%), tested positive. Five patients had local production of both anti-HSV and anti-VZV antibodies, whereas seven patients tested negative. Local antibody production was significantly associated with intraocular inflammation (P<0.05), corneal neovascularisation (P<0.05), and positive response to anti-viral treatment (P<0.05). No complications were encountered in sampling aqueous humor. CONCLUSIONS: Assessment of local anti-HSV and -VZV antibody production is a safe and reliable diagnostic procedure for recurrent herpes keratitis. It might be particularly helpful in patients presenting with intraocular inflammation and neovascularisation since it discriminates between herpes and non-herpes pathologies and may therefore be useful for preventive and therapeutic strategies. PMID- 16044330 TI - The functions of antennal mechanoreceptors and antennal joints in tactile discrimination of the honeybee (Apis mellifera L.). AB - Honeybees learn and discriminate excellently between different surface structures and different forms of objects, which they scan with their antennae. The sensory plate on the antennal tip plays a key role in the perception of mechanosensory and gustatory information. It is densely covered with small tactile hairs and carries a few large taste hairs. Both types of sensilla contain a mechanoreceptor, which is involved in the antennal scanning of an object. Our experiments test the roles of the mechanoreceptors on the antennal tip in tactile antennal learning and discrimination. Joints between head capsule and scapus and between scapus and pedicellus enable the bee to perform three-dimensional movements when they scan an object. The role of these joints in tactile antennal learning and discrimination is studied in separate experiments. The mechanoreceptors on the antennal tip were decisive for surface discrimination, but not for tactile acquisition or discrimination of shapes. When the scapus pedicellus joint or the headcapsule-scapus joint was fixed on both antennae, tactile learning was still apparent but surface discrimination was abolished. Fixing both scapi to the head capsule reduced tactile acquisition. PMID- 16044332 TI - Motor activity and trajectory control during escape jumping in the locust Locusta migratoria. AB - We investigated the escape jumps that locusts produce in response to approaching objects. Hindleg muscular activity during an escape jump is similar to that during a defensive kick. Locusts can direct their escape jumps up to 50 degrees either side of the direction of their long axis at the time of hindleg flexion, allowing them to consistently jump away from the side towards which an object is approaching. Variation in jump trajectory is achieved by rolling and yawing movements of the body that are controlled by the fore- and mesothoracic legs. During hindleg flexion, a locust flexes the foreleg ipsilateral to its eventual jump trajectory and then extends the contralateral foreleg. These foreleg movements continue throughout co-contraction of the hindleg tibial muscles, pivoting the locust's long axis towards its eventual jump trajectory. However, there are no bilateral differences in the motor programs of the left and right hindlegs that correlate with jump trajectory. Foreleg movements enable a locust to control its jump trajectory independent of the hindleg motor program, allowing a decision on jump trajectory to be made after the hindlegs have been cocked in preparation for a jump. PMID- 16044331 TI - Acetylcholine, GABA and glutamate induce ionic currents in cultured antennal lobe neurons of the honeybee, Apis mellifera. AB - The honeybee, Apis mellifera, is a valuable model system for the study of olfactory coding and its learning and memory capabilities. In order to understand the synaptic organisation of olfactory information processing, the transmitter receptors of the antennal lobe need to be characterized. Using whole-cell patch clamp recordings, we analysed the ligand-gated ionic currents of antennal lobe neurons in primary cell culture. Pressure applications of acetylcholine (ACh), gamma-amino butyric acid (GABA) or glutamate induced rapidly activating ionic currents. The ACh-induced current flows through a cation-selective ionotropic receptor with a nicotinic profile. The ACh-induced current is partially blocked by alpha-bungarotoxin. Epibatidine and imidacloprid are partial agonists. Our data indicate the existence of an ionotropic GABA receptor which is permeable to chloride ions and sensitive to picrotoxin (PTX) and the insecticide fipronil. We also identified the existence of a chloride current activated by pressure applications of glutamate. The glutamate-induced current is sensitive to PTX. Thus, within the honeybee antennal lobe, an excitatory cholinergic transmitter system and two inhibitory networks that use GABA or glutamate as their neurotransmitter were identified. PMID- 16044333 TI - [Perinatal risk in emergent surgery]. AB - During pregnancy, appendicitis, cholecystitis, choledocholithiasis, intestinal obstruction, and ulcers can lead to complaints and complications which occasion emergent surgery. The disease as such, its complications, the surgical intervention, and the anesthesia represent a risk for both mother and child. Risks arise particularly often due to the delay of diagnosis and therapy. Risk to the child depends on gestational week, fetal lung maturity, and oxygen supply. PMID- 16044334 TI - [Polytraumatized burn injury victims]. AB - Most polytraumatized burn injury persons aged 30-40 years who have suffered a burn or a burn-like injury are stigmatized by serious functional deficits, temporary loss of their social independence, injury-associated psychological reactions of different levels, and unusual reversible deficits in their higher cortical functions. The result is a prolonged stage of acute treatment and rehabilitation as well as very often the lifelong need for highly specialized care by physicians and other medical professionals. Accordingly, the deficits in any individual case are high and in spite of the low case rate (approximately 300 400 cases/ year) these accidents have a high impact on society as a whole, resulting in a significant financial burden for the social security system, insurance funds, and pension funds. PMID- 16044335 TI - [Emergency management of thoracic trauma]. AB - Thoracic injuries are a major cause of mortality during the "golden hour" of trauma. Many patients with chest trauma die after reaching the hospital. Less than 10% of all blunt thoracic injuries require a thoracotomy, and many potentially life-threatening conditions can be relieved by simple procedures, such as chest tube insertion.Thus, many cases of traumatic deaths due to chest injury may be prevented by prompt diagnosis and a standardized therapeutic approach in the emergency room. A high index of suspicion for lethal injury patterns, based on the mechanism of trauma and the clinical presentation, is a crucial prerequisite for an adequate initial assessment and management of patients with chest trauma. The worldwide implementation of standardized diagnostic and therapeutic guidelines, such as the "Advanced Trauma Life Support" (ATLS) protocol, has led to a significant reduction of early deaths attributed to thoracic injuries. PMID- 16044336 TI - [Surgical anatomy of the lumbar spine]. AB - The success and risks of spinal surgery depend on a correct diagnosis and detailed knowledge of anatomy. This report provides information on relevant structures of the lumbar spine and possible surgical complications. Anatomical topography and different surgical approaches are described step by step. PMID- 16044337 TI - [Subtalar screw-arthroereisis for correction of flat foot in children]. AB - Minimally invasive subtalar screw-arthroereisis has gained interest in the correction of flat feet in children. Between 1990 and 2004, this technique was used on 152 children, 74 bilaterally, for a total of 226 feet. There were 82 boys and 70 girls, with an average age of 10.6+/-1.9 years. The results were good in 95.4% of cases, whilst there were complications in 4.6%. For well corrected feet, 55 screws have now been removed, on average 2.9 years after implantation. The indications for such treatment are: talipes calcaneovalgus, which develops into a flat foot (spontaneous correction can be expected to 10-11 years), juvenile flat foot with medial protrusion of the talar head and complete absence of the longitudinal arch, symptomatic talipes calcaneovalgus with pain on the insertion of the tibialis posterior tendon, a minimum age of 6 years if a correction with conservative treatment does not show any improvement; a maximum age of 12-13 years considering that there always has to be a manual reducibility of the deformity. CONTRAINDICATIONS: posttraumatic flat foot, congenital flat foot, stiff flat foot, age over 13 years. PMID- 16044338 TI - Influence of fat pad removal on patellar tendon length during growth. AB - During various knee operations, the changes caused by the surgical invasion to the infrapatellar fat pad (IPF) is still unknown. If any changes exist, it will have great influence especially on growing generations. Eighty-four Japanese white rabbits (6-month-old) were divided into three groups: the resection group involving resection of the IPF, the graft group involving resection and reimplantation of the IPF, and the no-surgery group. All these surgical procedures were done in right knees. In all left knees, only arthrotomy was performed, serving as the sham side. After 3, 6, 12, and 24 weeks of the operation, the rabbits were killed. Lengths of the patellar tendon and patellar were measured in lateral X-ray. In order to eliminate individual differences in the patellar height, we defined a new index as percent patellar height (PPH) which indicated the percentage of the patellar height of surgery side compared with that of the sham side. The PPH was 90.6% (3 weeks), 83.0% (6 weeks), 73.6% (12 weeks), and 74.7% (24 weeks) in the resection group, while it was 88.4% (6 weeks), and 88.9% (24 weeks) in the graft group. Postsurgical scar tissue formation occurring where the IPF was removed prevented the normal growth of the patellar tendon. Reimplantation of the IPF lessened the adhesion of the patellar tendon to the surrounding tissue, and better growth of the tendon. These results showed that preservation of the IPF in young individuals could be crucial for the normal growth of the patellar tendon, and critical as well for the prevention of the degeneration of the articular surface. PMID- 16044339 TI - Statin-macrolide interaction risk: a population-based study throughout a general practice database. AB - INTRODUCTION: The myopathy risk associated with statin use increases in case of concomitant prescription of certain drugs, such as cytochrome P(450) (CYP) system inhibitors (i.e., macrolides). The aim of this study was to assess whether concurrent statin/macrolide prescriptions at high interaction risk are commonly written in a general practice setting. METHODS: For this study, 156 general practitioners (GPs) with a patient population of almost 200,000 individuals, and participating in the Arianna database set up by "Caserta-1" Local health-Service Agency (ASL), were recruited. Within such a study sample, subjects receiving at least one statin/macrolide concomitant prescription during the year 2003 were identified. Sensitivity analysis was performed to assess the time distribution of high-risk macrolide prescriptions written within +/-10 days from the statin prescription date. RESULTS: Among 190,124 patients included in the study, 7,176 (3.8%) received at least one statin prescription during the observation period. Of these, 228 (3.2%) were occasionally co-prescribed with any macrolide on the same date, in 153 cases (2.1% of statin users) the macrolide being of high interaction risk. In particular, 2.1% (55) of simvastatin users and 2.2% (64) of patients on atorvastatin were prescribed with high-risk macrolide on the same date versus 0.6% and 1.8% of patients prescribed with fluvastatin and pravastatin, respectively. Concerning GPs, 99 (63.5%) co-administered statin and macrolide at high interaction risk at least once. CONCLUSIONS: Most GPs occasionally prescribed statin/macrolide at high interaction risk on the same date, despite the availability of therapeutic alternatives. Prevention strategies targeted to increase awareness of health professionals about the interaction risks of widely prescribed drugs are needed. PMID- 16044340 TI - Fluoropyrimidine therapy: hyperbilirubinemia as a consequence of hemolysis. AB - BACKGROUND: Hemolytic anemia has been noted during treatment with a variety of chemotherapeutic agents. We observed mild compensated hemolytic anemia in a patient receiving capecitabine during a randomized, controlled trial of adjuvant therapy. In order to investigate the hypothesis that hemolysis is the underlying cause of the hyperbilirubinemia sometimes observed during capecitabine treatment, we evaluated factors associated with hemolysis in ten patients. Factors were also analyzed in ten patients receiving 5-fluorourocil/leucovorin (5-FU/LV). METHODS: Twenty chemotherapy-naive patients undergoing surgery for Dukes' C colon cancer were included in the phase III, 'X-ACT' trial, and randomized to receive 24-week adjuvant treatment with either oral capecitabine (eight cycles of 1,250 mg/m2 twice daily for 14 days, followed by a 7-day rest period) (n=10) or 5-FU/LV administered according to the Mayo Clinic regimen (six cycles of LV 20 mg/m2 followed by 5-FU 425 mg/m2, administered as an i.v. bolus on days 1-5 every 28 days) (n=10). Ten patients randomized in each treatment arm were evaluated. Hemolytic parameters evaluated included bilirubin, lactate dehydrogenase, haptoglobin, and reticulocytes. RESULTS: Seven patients receiving capecitabine and three patients receiving 5-FU/LV experienced grade 1/2 elevations of bilirubin during the 24-week treatment period. In most cases, hyperbilirubinemia was associated with concomitant alterations in other hemolytic parameters. Five episodes of grade 1 compensated hemolytic anemia were reported in four capecitabine-treated patients, all of which were associated with hyperbilirubinemia. CONCLUSION: Adjuvant treatment with capecitabine or 5-FU/LV in a small sample of patients with Dukes' C colon cancer was associated with alterations in hemolytic parameters. These alterations, in particular hyperbilirubinemia, were associated in some patients with low-grade compensated hemolytic anemia. All changes were clinically insignificant, fully reversible, and may represent a fluoropyrimidine class effect. Further studies are indicated to evaluate the incidence and implications of this effect. PMID- 16044341 TI - Role of Na+, K+-ATPase alpha1 subunit in the intracellular accumulation of cisplatin. AB - The present study was undertaken to identify what regulates intracellular cisplatin (CDDP) accumulation and what changes in membrane fraction of CDDDP resistant cell line. The CDDP-resistant rat hepatoma cell line, H4-II-E/CDDP, shows a significant decrease in intracellular platinum accumulation compared with parental H4-II-E cells, although there was no difference in the efflux of CDDP between these two cell lines. In this study, we examined the contribution of functional change in active transport to the CDDP resistance of H4-II-E/CDDP cells. Compared with the resistant cells, platinum accumulation in the parental cells was clearly decreased by low temperature or ATP depletion. In addition, the Na+, K+-ATPase inhibitor ouabain and the K+ channel inhibitor tetraethylammonium decreased platinum accumulation in parental cells but did not change the accumulation in resistant cells. Amphotericin B, an antifungal agent, increased the intracellular platinum accumulation in resistant cells to the same level as in parent cells. Western blot analysis demonstrated that the Na+, K+-ATPase alpha1 subunit was reduced in resistant cells compared with the parental cells, although there was no difference in the expression of the beta1 subunit between the two cell lines. Furthermore, the Na+, K+-ATPase alpha1 subunit of H4-II-E was decreased following a 24-h exposure to CDDP. These results suggest that Na+, K+ ATPase-dependent active transport of CDDP does not occur in resistant cells, and, furthermore, our findings provide the first evidence that the Na+, K+-ATPase alpha1 subunit plays an important role in the transport of CDDP. PMID- 16044342 TI - A history of the treatment of craniopharyngiomas. AB - RATIONALE: Nearly 100 years have passed since the first surgical attempt to remove a craniopharyngioma was undertaken, and the management of patients with these tumors continues to both challenge and frustrate those involved with their treatment. METHODS: By constructing a historical framework from past literature on the treatment of craniopharyngioma, the authors have attempted to document the early and important events in the treatment of these tumors in order to add a historical dimension to their current treatment and to engender a more comprehensive understanding of the modern treatment of craniopharyngioma. CONCLUSION: The application of radiotherapeutic techniques to the management of craniopharyngioma would begin to complement the role of surgery, but debate would soon emerge as to what constituted the most appropriate management for craniopharyngioma in children. PMID- 16044343 TI - Craniopharyngiomas in children: surgical experience at Children's Memorial Hospital. AB - OBJECTIVES: Craniopharyngioma during childhood poses difficulty in management because of the high incidence of surgical complications and treatment failure. In order to identify less detrimental and more effective treatment, a personal series of craniopharyngioma was reviewed in regard to various clinical factors, patient factors (age and sex), tumor factors (location and extension, relationship with chiasm, and hydrocephalus), and therapeutic modes [extent of resection and radiation therapy (RT)]. MATERIALS AND METHODS: Fifty-four childhood craniopharyngiomas treated from 1984 to 2003 were reviewed. Preoperative neuroimaging studies were classified depending upon tumor location and extension. In this series of 54 patients, 43 had total tumor resection and 11 had subtotal resection. Of the total resection group, ten showed evidence of residual tumor on postoperative neuroimaging studies. Following the initial resection, 46 did not have RT whereas 8 with subtotal resection received RT. RESULTS: There were no surgical deaths. Postoperative complications included pseudoaneurysm in 1, hemiparesis in 3, severe obesity in 5, panhypopituitarism in 50, and worsening of visual function in 7. During follow-up ranging from 12 months to 21 years, 24 patients had recurrence. Of the 33 patients with radiographic total resection, 9 (27.3%) had recurrence. Among the patients with total resection but radiographic residual and those with subtotal resection, the craniopharyngioma recurred in 90% and 100%, respectively. Three (37.5%) of eight patients with subtotal resection with RT had recurrence. Overall recurrence-free survival was 62% at 5 years and 49% at 10 years. The sex and age, location and extension of the tumor, nature of the optic chiasm, and hydrocephalus did not influence survival with statistical significance. However, the extent of surgical resection and use of RT showed significant differences for survival. Patients with total resection had a recurrence-free survival rate of 83% and 70% at 5 and 10 years, respectively. Patients with subtotal resection with RT had 71% at 5 years and 36% at 10 years. Patients who had subtotal resection or radiographically residual tumor without RT had a recurrence-free survival rate of only 9%. Among 22 patients whose recurrent tumor was treated with RT, a second recurrence-free survival rate was 90% at 5 years. CONCLUSION: Total resection provided the best outcome. However, recurrence rates and surgical complications remained high following radical tumor resection. RT was effective for recurrent tumors and should be considered being the primary treatment for recurrences or difficult tumors, which are not amenable to total resections. PMID- 16044344 TI - Epidemiology of pediatric tumors of the nervous system according to the WHO 2000 classification: a report of 1,195 cases from a single institution. AB - OBJECT: The purpose of this study is to determine the epidemiology of tumors of the nervous system diagnosed according to the WHO 2000 classification in a single Brazilian institution. PATIENTS AND METHOD: One thousand one hundred ninety-five tumors in children between 0 and 21 years of age diagnosed between 1974 and 2003 were classified according the sex, topography, and age distribution. RESULTS: In all ages, males were slightly more affected. In the first 2 years, the prevalence for boys was higher (68.3%). In the whole series, 58.7% were supratentorial, 31.4% infratentorial, and 9.9% spinal (44% intra- and 56% extramedullary). Among these latter, ependymomas and schwannomas were the most frequent. In the cerebral compartment, pilocytic astrocytomas were the single most frequent tumors (18%), followed by diffuse astrocytomas (14%), medulloblastomas (11%), and craniopharyngiomas (11%). In the posterior fossa, there was an even distribution among medulloblastomas and pilocytic astrocytomas, but the former was much more frequent in the first 2 years of age. High-grade (III and IV) diffuse astrocytomas were slightly more frequent than low grades (II), and this difference becomes more evident as the child grows older. Due to the new development of the surgery of epilepsy, the frequency of neuronal and mixed neuronal-glial tumors is increasing (8%). CONCLUSION: Classified according to the latest WHO classification, by a single neuropathologist in a single institution, this large series of pediatric neurological tumors may reflect fairly well their real incidence. Our results obtained in a developing country do not differ substantially from other similar series reported in the literature from the First World. PMID- 16044345 TI - Gemcitabine as prolonged infusion and vinorelbine in anthracycline and/or taxane pretreated metastatic breast cancer: a phase II study. AB - BACKGROUND: Gemcitabine and vinorelbine are active agents for the treatment of metastatic breast cancer. Prolonged infusion of gemcitabine can result in higher levels of active metabolites compared to shorter administration. This phase II trial was initiated to evaluate the efficacy and tolerability of gemcitabine as prolonged infusion in combination with vinorelbine in anthracycline and/or taxane pretreated patients with metastatic breast cancer. PATIENTS AND METHODS: Patients who had received one prior line of chemotherapy for metastatic disease were treated with gemcitabine (350 mg/m2 as 4 h infusion) and vinorelbine (25 mg/m2 on days 1 and 8. Treatment was repeated every 3 weeks for a maximum of six cycles. RESULTS: Of 26 patients enrolled, 84% had received prior anthracycline treatment and 50% prior taxane therapy. In total, one complete and six partial responses were achieved, accounting for an overall response rate of 30.4%. The clinical benefit rate was 47.8%. Median duration of response and median time to progression were 7.3 months and 4.6 months, respectively. Median overall survival was 14.5 months. Although the predominant toxicity was myelosuppression with grade 3/4 neutropenia in 42% of patients, few neutropenic complications resulted. Non-hematological toxicity was generally moderate. Most common non-hematologic toxicities were nausea, vomiting, alopecia, peripheral neuropathy and elevation of liver enzymes. CONCLUSION: Gemcitabine as prolonged infusion and vinorelbine are a safe and effective combination treatment in anthracycline and/or taxane pretreated patients. Approximately 47.8% of patients derived clinical benefit from treatment. This regimen represents a therapeutic option for patients receiving second-line therapy for metastatic breast cancer. PMID- 16044346 TI - Prognostic significance of VEGF immunohistochemical expression and tumor angiogenesis in head and neck squamous cell carcinoma. AB - PURPOSE: Tumor angiogenesis is crucial for both the growth of the primary tumor and the development of metastases. Among the factors causing tumor angiogenesis, vascular endothelial growth factor (VEGF) is considered as a leading candidate. We aimed to assess the prognostic significance of VEGF and tumor angiogenesis in head and neck squamous cell carcinoma (HNSCC). METHODS: We performed a retrospective study of 69 patients with HNSCC, in order to investigate whether VEGF immunohistochemical expression and tumor angiogenesis correlate with clinicopathological parameters and outcome. Tumor angiogenesis was estimated by determining microvessel density (MVD), and VEGF expression was assessed quantitatively. RESULTS: Vascular endothelial growth factor and MVD correlated statistically significant with the clinical stage, but not with the presence of lymph node metastasis at the time of diagnosis. Tumors located in the oral cavity and larynx more often expressed high VEGF immunostaining compared with tumors located in the lower lip. High VEGF expression was associated with higher clinical stage and worse overall survival in this cohort of patients. CONCLUSIONS: Vascular endothelial growth factor expression may have prognostic significance for patients with HNSCC. PMID- 16044347 TI - Quantitation of synergism of arabinosylcytosine and cladribine against the growth of arabinosylcytosine-resistant human lymphoid cells. AB - This report presents a quantitative analysis of the synergistic interaction of arabinosylcytosine (araC) and cladribine (CdA) in human H9-lymphoid cell lines sensitive and resistant to araC (H9-araC cells). H9-araC cells obtained by cultivation of H9 cells in the presence of 0.5 microM arabinosylcytosine (araC) had lower deoxycytidine kinase (dCK) than the parental cell line. The IC50 values of araC and CdA calculated by using median-effect analysis and CalcuSyn software were: 0.55 microM and 1.16 microM for CdA and 0.0058 microM and 3.5 microM for araC in H9 and H9-araC cells, respectively. These values were reduced to 0.10 microM and 0.38 microM for CdA and to 0.004 microM and to 0.77 microM for araC when the drugs were used in combination. Computerized simulation of dose reduction index (DRI) indicated that at 50-99% growth inhibition levels, the doses of araC could be reduced by 2.0 to 11.9-fold and 2.9 to 5.3-fold and the doses of CdA by 5.9 and 183.7-fold and 3.1 to 164.8-fold in H9 and H9-araC cells, respectively, when the drugs are used in combination. Assessment by combination index (CI) analysis showed that the combination exhibited moderate to strong synergistic lympho-cytotoxic effects. CdA metabolic studies (influx and activation) in the presence of deoxyadenosine, deoxycytidine, or araC suggested that CdA enters cells by a deoxyadenosine-inhibitable transport system, which is different than that of araC and deoxycytidine transport system. Thus, in addition to the known mechanisms, other mechanisms might be involved in the metabolism of CdA. The demonstration that araC and CdA combinations exert synergistic cytotoxicity even in the resistant cells raises hope that such a combination may be useful in tumors that were found resistant to these drugs. PMID- 16044348 TI - Meteorology and the physical activity of the elderly: the Nakanojo Study. AB - Seasonal changes in ambient temperature and day length are thought to modify habitual physical activity. However, relationships between such environmental factors and the daily physical activity of older populations remain unclear. The present study thus examined associations between meteorological variables and the number of steps taken per day by elderly Japanese. Continuous pedometer counts over a 450-day period were collected from 41 healthy subjects (age 71+/-4 years), none of whom engaged in any specific occupational activity or exercise programs. An electronic physical activity monitor was attached to a belt worn on the left side of the body throughout the day. Daily values for mean ambient temperature, duration of bright sunshine, mean wind speed, mean relative humidity, and precipitation were obtained from local meteorological stations. The day length was calculated from times of sunrise and sunset. Based on the entire group of 41 subjects (ensemble average), a subject's step count per day decreased exponentially with increasing precipitation (r2=0.19, P<0.05). On days when precipitation was <1 mm, the step count increased with the mean ambient temperature over the range of -2 to 17 degrees C, but decreased over the range 17 29 degrees C. The daily step count also tended to increase with day length, but the regression coefficient of determination attributable to step count and mean ambient temperature (r2=0.32, P<0.05) exceeded that linking the step count and day length (r2=0.13, P<0.05). The influence of other meteorological factors was small (r2 or = 50 years reported a recent colorectal cancer screening test, and 42% reported a colonoscopy within the past 10 years. After multiple statistical adjustments, groups with the lowest likelihood of screening were the poor (odds ratio [OR], 0.66; 95% confidence interval [CI], 0.53-0.83) and uninsured (OR, 0.31; 95% CI, 0.20-0.48), as well as Asians (OR, 0.46; 95% CI, 0.29-0. 72), and current smokers (OR, 0.62; 95% CI, 0.50-0.78). Colonoscopy was less frequently reported by non-Hispanic Black New Yorkers and by women; both groups reported higher use of fecal occult blood tests. Less than 10% of adult New Yorkers reported a sigmoidoscopy in the past 5 years. CONCLUSIONS: Low screening uptake in NYC leaves nearly 1 million New Yorkers, particularly poor and uninsured adults, at risk for undetected colorectal cancer. Colonoscopy screening programs in NYC should address health care and socioeconomic barriers and target racial and ethnic minorities and women. PMID- 16044402 TI - Alpha-1-antitrypsin deficiency: a new paradigm for hepatocellular carcinoma in genetic liver disease. AB - Liver disease in alpha-1-antitrypsin (alpha1AT) deficiency is caused by a gain-of toxic function mechanism engendered by the accumulation of a mutant glycoprotein in the endoplasmic reticulum (ER). The extraordinary degree of variation in phenotypical expression of this liver disease is believed to be determined by genetic modifiers and/or environmental factors that influence the intracellular disposal of the mutant glycoprotein or the signal transduction pathways that are activated. Recent investigations suggest that a specific repertoire of signaling pathways are involved, including the autophagic response, mitochondrial- and ER caspase activation, and nuclear factor kappaB (NFkappaB) activation. Whether activation of these signaling pathways, presumably to protect the cell, inadvertently contributes to liver injury or perhaps protects the cell from one injury and, in so doing, predisposes it to another type of injury, such as hepatocarcinogenesis, is not yet known. Recent studies also suggest that hepatocytes with marked accumulation of alpha1ATZ, globule-containing hepatocytes, engender a cancer-prone state by surviving with intrinsic damage and by chronically stimulating in 'trans' adjacent relatively undamaged hepatocytes that have a selective proliferative advantage. Further, this paradigm may apply to other genetic and infectious liver diseases that are predisposed to hepatocellular carcinoma. PMID- 16044403 TI - Simple surface modification of poly(epsilon-caprolactone) to induce its apatite forming ability. AB - A biodegradable polymer coated with a bone-like apatite layer on its surface is useful as a scaffold for bone tissue regeneration. In this work, a poly(epsilon caprolactone) (PCL) surface was modified by an O2 plasma surface treatment to form oxygen-containing functional groups. The plasma-treated samples were subsequently dipped alternately in an alcoholic solution containing calcium ions and one containing phosphate ions to deposit apatite precursors on the surface. The surface-modified PCL samples formed a dense and uniform surface bone-like apatite layer after immersion for 24 h in a simulated body fluid with ion concentrations approximately equal to those of human blood plasma. This surface modification process is applicable to two-dimensional PCL plates and three dimensional PCL meshes. In the resulting apatite-PCL composite, the apatite layer strongly adhered to the PCL surface and remained intact after a tape-detachment test. Therefore, this type of composite material will be a useful scaffold for bone tissue engineering. PMID- 16044404 TI - Preparation and characterization of nano-hydroxyapatite/chitosan composite scaffolds. AB - A novel nano-hydroxyapatite (HA)/chitosan composite scaffold with high porosity was developed. The nano-HA particles were made in situ through a chemical method and dispersed well on the porous scaffold. They bound to the chitosan scaffolds very well. This method prevents the migration of nano-HA particles into surrounding tissues to a certain extent. The morphologies, components, and biocompatibility of the composite scaffolds were investigated. Scanning electron microscopy, porosity measurement, thermogravimetric analysis, X-ray diffraction, X-ray photoelectron spectroscopy, and Fourier transformed infrared spectroscopy were used to analyze the physical and chemical properties of the composite scaffolds. The biocompatibility was assessed by examining the proliferation and morphology of MC 3T3-E1 cells seeded on the scaffolds. The composite scaffolds showed better biocompatibility than pure chitosan scaffolds. The results suggest that the newly developed nano-HA/chitosan composite scaffolds may serve as a good three-dimensional substrate for cell attachment and migration in bone tissue engineering. PMID- 16044405 TI - Differential gene expression in epidermis of mice sensitive and resistant to phorbol ester skin tumor promotion. AB - Previous data from two-stage carcinogenesis studies in mouse skin demonstrated that genetic control of susceptibility to skin tumor promotion by the phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA), in crosses between susceptible DBA/2J and resistant C57BL/6J mice is a multigenic trait. Utilizing a cDNA microarray approach, we compared global gene expression profiles in the epidermis of these two mouse strains treated with TPA or vehicle (acetone). Gene expression in the epidermis was analyzed after the treatment to identify global effects of TPA, as well as potential candidate genes that modify susceptibility to skin tumor promotion. DBA/2J and C57BL/6J mice were treated topically four times with 3.4 nmol TPA or acetone over a 2-wk period, and RNA was extracted from epidermis 6 h after the final treatment. Labeled cDNA generated from each group was hybridized to commercial cDNA microarrays (Agilent) containing more than 8000 targets. More than 450 genes were significantly influenced, directly or indirectly, by TPA treatment in the epidermis of either strain. Notably, 44 genes exhibited differential expression between the tumor promotion sensitive and resistant mouse strains. Several genes that were differentially expressed in DBA/2J versus C57BL/6J epidermis after TPA treatment were located in chromosomal regions linked to TPA promotion susceptibility. Three genes, Gsta4, Nmes1 (MGC58382), and Serpinb2, located within promotion susceptibility loci Psl1 (chr 9), Psl2 (chr 2), and Psl3 (chr 1), respectively, were identified in this analysis as potential candidates for modifiers of susceptibility to skin tumor promotion by TPA. PMID- 16044406 TI - Improved performance of primary rat hepatocytes on blended natural polymers. AB - Alginate (Alg), chitosan (Chi), collagen (Col), gelatin (Gel), and the mixtures of every two of them were screened to determine their suitability for hepatocyte culture. The test materials were fabricated as films and then evaluated on the basis of their abilities to promote the attachment and functions of rat hepatocytes cultured on them. Cellular attachment on Col and Gel was favorable. However, cellular viability, cytoskeleton organization and function, as evaluated by albumin production, ureagenesis, and enzyme activity of cytochrome P450 as well as expression levels of hepatocyte nuclear factor 4 alpha deteriorated. Reverse cellular behavior was observed on Alg and Chi. Two blends, composed of Chi and Col or Gel, were found to be superior to other materials and sustained viability and differentiated functions of hepatocyte. PMID- 16044407 TI - Inhibition of TPA-induced cyclooxygenase-2 (COX-2) expression by apigenin through downregulation of Akt signal transduction in human keratinocytes. AB - Apigenin is a nonmutagenic bioflavonoid that has been shown to be an inhibitor of mouse skin carcinogenesis induced by the two-stage regimen of initiation and promotion with dimethylbenzanthracene (DMBA) and 12-O-tetradecanoylphorbol-13 acetate (TPA). These DMBA/TPA-induced squamous cell carcinomas overexpress cyclooxygenase-2 (COX-2). Cyclooxygenases are key enzymes required for prostaglandin (PG) synthesis, converting the arachidonic acid (AA) released by phospholipase A2 into prostaglandins. A large body of evidence indicates that the inducible form of cyclooxygenase, COX-2, is involved in tumor promotion and carcinogenesis in a wide variety of tissue types, including colon, breast, lung, and skin. In the present study, we have determined that apigenin inhibited the TPA-induced increase in COX-2 protein and mRNA in the human keratinocyte cell line; HaCaT. The induction of COX-2 elicited by TPA correlated with increased activation of Akt kinase and cell treatment with the PI3 kinase inhibitor, LY294002, blocked TPA induction of COX-2. In cells treated with TPA and apigenin, the inhibition of COX-2 expression correlated with inhibition of Akt kinase activation. Apigenin-mediated inhibition of TPA-induced COX-2 expression was reversed by transient transfection with constitutively active Akt (CA-Akt). Chemical inhibitors of MEK (PD98059), p38 (SB202190), but not JNK (SP600125) blocked TPA induction of COX-2 although apigenin did not inhibit TPA-mediated COX 2 expression through these pathways. The TPA-induced release of AA from HaCaT cells was also inhibited by cell treatment with apigenin. These data show that apigenin inhibits TPA-mediated COX-2 expression by blocking signal transduction of Akt and that apigenin also blocks AA release, which may contribute to its chemopreventive activity. PMID- 16044408 TI - Comparison of the effects of nitric oxide donors and calcium channel blockers on the intrinsic myogenic tone of sheep isolated internal anal sphincter. AB - BACKGROUND: Chronic anal fissure is associated with considerable pain and anal hypertonia. Numerous clinical studies attest to the effectiveness of individual nitro-containing drugs and organic calcium channel blockers in this condition but there are few comparative studies. METHODS: Isolated segments of sheep internal anal sphincter were prepared for isometric tension recording. The effect of various drugs on myogenic tone was examined in the absence or presence of sodium orthovanadate (SOV), an agent used to mimic anal hypertonia by increasing myogenic tone. RESULTS: All the drugs tested produced concentration-dependent inhibition of myogenic tone, with the maximum effect ranging from 66.4 per cent (verapamil) to 100 per cent (sodium nitroprusside). Sodium nitroprusside and diltiazem were the most potent, followed by glyceryl trinitrate (GTN), nifedipine and verapamil, which had similar potency, and finally nicorandil. The potency of GTN and diltiazem was reduced threefold in the presence of 1 mmol/l SOV. The combined effect of GTN and diltiazem was greater than the effect of either agent alone, even in the presence of 3 mmol/l SOV. CONCLUSION: Nitro-containing drugs and organic calcium channel blockers are potent inhibitors of anal sphincter myogenic tone that may be used in combination to treat chronic anal fissure. PMID- 16044409 TI - Percutaneous thermal ablation for recurrent hepatocellular carcinoma after hepatectomy. AB - BACKGROUND: Tumour ablation using a thermal energy source has shown promising results, and is particularly suitable for recurrent hepatocellular carcinoma (HCC). The present study evaluated long-term outcomes after percutaneous thermal ablation for recurrent HCC following liver resection. METHODS: Radiofrequency ablation or microwave ablation was used to treat a total of 124 tumour nodules (0.9-7.0 cm in diameter) in 72 patients with recurrent HCC. RESULTS: Complete ablation of 119 (96.0 per cent) of 124 tumour nodules was achieved. There was no treatment-related death and the major complication rate was 4 per cent. During a mean(s.d.) follow-up of 27.9(17.8) months, local recurrence developed in 16 (13.6 per cent) of 118 successfully treated tumour nodules. Distant recurrence developed in 60 (85 per cent) of 71 patients, of whom 26 had repeat metachronous distant recurrence. With repeated ablation for both local and distant recurrence, the 1-, 3- and 5-year overall survival rates after initial ablation were 75, 43 and 18 per cent respectively. Patients with a serum alpha-fetoprotein level greater than 200 ng/ml before treatment had significantly poorer survival than those with a lower level (P = 0.034) and multivariate analysis identified preablation AFP level as an independent prognostic factor (P = 0.054). CONCLUSION: With their advantages of preservation of non-tumorous liver tissue and easy repetition, percutaneous thermal ablative therapies were particularly suitable for recurrent HCC and improved long-term survival. PMID- 16044410 TI - Meta-analysis of the value of somatostatin and its analogues in reducing complications associated with pancreatic surgery. AB - BACKGROUND: The role of somatostatin and its analogues in reducing complications after pancreatic resection is controversial. This is a meta-analysis of the evidence of benefit. METHODS: A literature search using Medline and ISI Proceedings with exploration of the references identified 22 studies. Of these, ten met the inclusion criteria for data extraction. Estimates of effectiveness were performed using fixed- and random-effects models. The effect was calculated as an odds ratio (OR) with 95 per cent confidence intervals (c.i.) using the Mantel-Haenszel method. Level of significance was set at P < 0.050. RESULTS: Outcomes for 1918 patients were compared. Somatostatin and its analogues did not reduce the mortality rate after pancreatic surgery (OR 1.17 (0.70 to 1.94); P = 0.545) but did reduce both the total morbidity (OR 0.62 (0.46 to 0.85); P = 0.003) and pancreas-specific complications (OR 0.56 (0.39 to 0.81); P = 0.002). Somatostatin and its analogues reduced the rate of biochemical fistula (OR 0.45 (0.33 to 0.62); P < 0.001) but not the incidence of clinical anastomotic disruption (OR 0.80 (0.44 to 1.45); P = 0.459). CONCLUSION: Somatostatin and its analogues reduce the incidence of complications after surgery. PMID- 16044411 TI - Aggressive resection of the airway invaded by thyroid carcinoma. AB - BACKGROUND: The aim of this study was to investigate the hypothesis that outcome following concomitant airway resection is superior to that after shaving of the tumour in patients with airway invasion of thyroid carcinoma. METHODS: The records of 34 patients with thyroid cancer with airway invasion were reviewed retrospectively. In addition to total thyroidectomy, airway resection was performed in 18 patients (group 1), whereas the tumour was shaved away from the airway in the other 16 patients (group 2). 131I was used as postoperative adjuvant therapy in all patients. Metastasis and recurrence of the primary lesion were determined by 131I whole-body scans, serum thyroglobulin levels, and computed tomography or ultrasonography of the neck. RESULTS: In group 1, two anastomotic dehiscences resulted in one death. Patients in group 2 had a higher rate of local recurrence (relative risk 8.0, P = 0.013) and earlier recurrence (mean(s.e.m.) 2.6(0.8) versus 7.0(1.1) years; P = 0.026) than those in group 1. Median survival was 5.8 and 4.3 years in the 18 patients of group 1 and 16 patients of group 2 (P = 0.259), and the respective 5-year survival rates were 88 and 84 per cent (P = 0.783). CONCLUSION: Aggressive airway resection can minimize local recurrence of thyroid carcinoma with airway invasion. PMID- 16044412 TI - Evaluation of treatment and long-term follow-up in patients with hepatic alveolar echinococcosis. AB - BACKGROUND: Alveolar echinococcosis is a rare disorder, which makes a comparison of different treatment modalities within a clinical trial difficult to perform. Data prospectively recorded over a period of 25 years were used to evaluate three therapeutic strategies: benzimidazole therapy alone, complete 'curative' resection followed by 2 years of adjuvant benzimidazole treatment, and partial debulking resection followed by continuous administration of a benzimidazole. METHODS: Details of 113 patients with hepatic alveolar echinococcosis treated between 1976 and 2003 were analysed. Kaplan-Meier survival curves were constructed and, using a Cox regression model, patient age, year of initial treatment and PNM stage were entered as co-variates in the analysis. RESULTS: Kaplan-Meier overall survival curves stratified for treatment strategy indicated an improved long-term survival in patients undergoing the debulking procedure (P = 0.061) or curative resection (P = 0.002) compared with benzimidazole therapy alone. However, when PNM stage, patient age and year of initial treatment were introduced into the analysis, there was a trend for survival advantage only with curative resection (P = 0.07 versus benzimidazole alone). Debulking resulted in a higher rate of progression of hepatic echinococcosis than curative surgery (P = 0.008). The incidence of parasite-related complications was similar for debulking resection and benzimidazole therapy alone (P = 0.706). CONCLUSION: Debulking hepatic resections do not appear to offer any advantage in the treatment of patients with alveolar echinococcosis. PMID- 16044413 TI - In vitro release of clomipramine HCl and buprenorphine HCl from poly adipic anhydride (PAA) and poly trimethylene carbonate (PTMC) blends. AB - Controlled drug-delivery technology is concerned with the systematic release of a pharmaceutical agent to maintain a therapeutic level of the drug in the body for modulated and/or prolonged periods of time. This may be achieved by incorporating the therapeutic agent into a degradable polymer vehicle, which releases the agent continuously as the matrix erodes. In this study, poly trimethylene carbonate (PTMC), an aliphatic polycarbonate, and poly adipic anhydride (PAA), an aliphatic polyanhydride, were synthesized via melt condensation and ring-opening polymerization of trimethylene carbonate and adipic acid, respectively. The release of clomipramine HCl and buprenorphine HCl from discs prepared with the use of PTMC-PAA blends in phosphate buffer (pH 7.4) are also described. Clomipramine HCl and buprenorphine HCl were both used as hydrophilic drug models. Theoretical treatment of the data with the Peppas model revealed that release of clomipramine HCl (5%) in devices containing 70% PTMC or more followed a Fickian diffusion model. However, the releases of buprenorphine HCl (5%) in the same devices were anomalous. For devices containing 50% and more PAA, surface erosion may play a significant role in the release of both molecules. PMID- 16044415 TI - Current trends in the structure-activity relationship studies of the endogenous agouti-related protein (AGRP) melanocortin receptor antagonist. AB - Agouti-related protein (AGRP) is an endogenous antagonist of the melanocortin-3 and -4 (MC3R and MC4) G-protein coupled receptors. The 87-132 amino acid C terminal domain of hAGRP possesses five disulfide bridges and a well-defined three-dimensional structure that displays full biological activity as compared to the full-length protein. Based on the NMR structure of the C-terminal AGRP(87 132), a novel mini-protein, referred to as "Mini-AGRP" was designed that exhibited receptor binding affinity and antagonism similar to that of the parent hAGRP(87-132) protein. It was demonstrated that this new-engineered protein autonomously folds to the inhibitor cystine knot (ICK) motif. As this AGRP is a novel mammalian protein involved in energy homeostasis and possibly other physiological functions remaining to be identified, structure-function studies are starting to emerge toward the understanding of how this unique protein putatively interacts with the melanocortin receptors with the objective of designing potential therapeutic agents for in vivo physiological studies. This article summarizes the progress to date of AGRP-based structure-activity relationships and putative ligand-receptor interactions. PMID- 16044414 TI - Transforming growth factor-beta 1 (TGF-beta1) prevents the age-dependent decrease in bone formation in human osteoblast/implant cultures. AB - Titanium implants have been extensively used in orthopedic surgery and dentistry. Most of the patients who receive such implants are elderly with a compromised ability to heal and form new bone. By using an in vitro osteoblast/implant culture system, the potency of TGF-beta1 in enhancing mineralization of human osteoblast cultures from elderly subjects was investigated in this study. Primary human osteoblast (HOB) cells obtained from different age group human subjects [Young (Y), Middle (M), and Old (O)] were cultured on Ti alloy (Ti-6Al-4V) disks with or without continuous administration of 0.2 ng/mL TGF-beta1 in the medium for 2 or 4 weeks. TGF-beta1 significantly (p < 0.05) increased calcium content and the size of calcified nodules on implant disks in the O group, but had no effect on the Y or M groups. The number of calcified nodules was not different with or without TGF-beta1 in all age groups. As measured by Northern blot analysis and RT-PCR, TGF-beta1 significantly increased the expression of bone specific extracellular matrix proteins, including alkaline phosphatase, Type I collagen, bone sialoprotein and osteocalcin, after both 2 and 4 weeks in the O group but not in the Y group. In conclusion, TGF-beta1 enhances mineralization on implant materials of osteoblast cultures from elderly human subjects. PMID- 16044416 TI - In vitro evaluation of platelet reactivity toward annuloplasty devices treated with heparin coating: studies under flow conditions. AB - We have applied an in vitro perfusion model to explore the potential thrombogenicity of polyester annulolasty fabric used in valve repair and to investigate the possible thromboresistance characteristics conferred by a special heparin coating (Duraflotrade mark treatment). Samples of human blood from i) untreated or ii) heparin-coated extracorporeal circuits were recirculated through annular perfusion chambers containing a) untreated or b) treated annuloplasty cloth material. Perfusion experiments were performed at a shear rate of 600 s(-1) for 20 min. Platelet interaction with the material was morphometrically evaluated. In experiments performed with blood from untreated circuits and cloth material, the average cross-sectional area of platelet mass was 615 +/- 135 microm2. Treatment of cloth material with Duraflotrade mark statistically decreased the area of interacting platelets to 319 +/- 101 microm2 (*p < 0.05, n = 10). Blood samples from heparin-coated extracorporeal circuits showed a decrease of total area of platelets (308 +/- 58 microm2 vs 138 +/- 30 microm2, *p < 0.05, n = 9). The combined treatment of Duraflotrade mark in extracorporeal circuits and cloth material caused a more consistent reduction (p < 0.05). The in vitro perfusion experimental model was sensitive to evaluate the thrombogenic potential of Duraflotrade mark treatment. Our results indicate that the heparin coating of cloth material and extracorporeal circuits improves the biocompatibility of the original material and reduces the thrombogenic profile. PMID- 16044417 TI - Selective inhibition of type X collagen expression in human mesenchymal stem cell differentiation on polymer substrates surface-modified by glow discharge plasma. AB - Recent evidence indicates that a major drawback of current cartilage- and disc tissue engineering is that human mesenchymal stem cells (MSCs) rapidly express type X collagen-a marker of chondrocyte hypertrophy associated with endochondral ossification. Some studies have attempted to use growth factors to inhibit type X collagen expression, but none to date has addressed the possible effect of the substratum on chondrocyte hypertrophy. Here, we sought to examine the growth and differentiation potential of human MSCs cultured on two polymer types, polypropylene and nylon-6, both of which have been surface-modified by glow discharge plasma treatment in ammonia gas. Cultures were performed for up to 14 days in Dulbecco's modified Eagle medium + 10% fetal bovine serum. Commercial polystyrene culture dishes were used as control. Reverse transcriptase-polymerase chain reaction was used to assess the expression of types I, II, and X collagens and aggrecan using gene-specific primers. Glyceraldehyde-3-phosphate dehydrogenase was used as a housekeeping gene. Types I and X collagens, as well as aggrecan, were found to be constitutively expressed by human MSCs on polystyrene culture dishes. Whereas both untreated and treated nylon-6 partially inhibited type X collagen expression, treated polypropylene almost completely inhibited its expression. These results indicate that plasma-treated polypropylene or nylon-6 may be a suitable surface for inducing MSCs to a disc like phenotype for tissue engineering of intervertebral discs in which hypertrophy is suppressed. PMID- 16044418 TI - Inhibition of the Akt, cyclooxygenase-2, and matrix metalloproteinase-9 pathways in combination with androgen deprivation therapy: potential therapeutic approaches for prostate cancer. AB - Prostate cancer cells are generally dependent on androgen stimulation mediated by the androgen receptor (AR) for growth and survival, and, therefore, hormonal manipulation, such as castration and/or the use of AR antagonists, results in a regression of the cancer. However, this treatment very rarely leads to the "cure" of advanced disease, and cancers eventually become androgen-independent. A number of genes/pathways have been reported to be activated in prostate cancer, most of which are possibly associated with disease progression. In this article, among them, we focus on Akt (also known as protein kinase B), cyclooxygenase (COX)-2, and matrix metalloproteinase (MMP)-9, whose activities or expressions have been found to be regulated by androgens/AR. Previous studies by us and others, with androgen-sensitive prostate cancer cell lines, have demonstrated that androgen deprivation results in activation/overexpression of Akt, COX-2, and MMP-9 in cells. This suggests that androgen deprivation in clinical settings activates the Akt, COX-2, and MMP-9 pathways in prostate cancer, which may increase cell growth and in turn promote the transition to the androgen-independent state. We hypothesize that androgen deprivation, in combination with inhibition of the Akt, COX-2, and MMP-9 pathways, delays the androgen-independent transition and has more beneficial effects than hormonal therapy alone. PMID- 16044419 TI - Biological and chemical inhibitors of NF-kappaB sensitize SiHa cells to cisplatin induced apoptosis. AB - Cisplatin, a chemotherapeutic agent, is known to induce apoptosis of cancer cells. We examined the role of NF-kappaB during cisplatin-induced apoptosis in two human cervical cancer cell lines, HeLa and SiHa, known to differ in their response to cisplatin treatment. We found that SiHa cells were relatively more resistant than HeLa cells to the cytotoxic effects induced by cisplatin as measured by MTT assays. HeLa cells were more sensitive to the apoptotic effects induced by cisplatin as shown by increases in annexin staining, DNA fragmentation, and loss of mitochondrial membrane potential. Similarly the activities of caspases 3, 8, and 9 and cleavage of PARP induced by cisplatin were more in HeLa than SiHa cells. Cisplatin induced NF-kappaB DNA binding activity in HeLa and SiHa cells but not in primary cervical cells and the active DNA binding complex in SiHa cells consists of p50 and RelA heterodimers. However, when NF kappaB DNA binding activity was blocked by chemical (curcumin, PDTC, or salicylic acid) or biological inhibitors (NIK-KM or IKK-beta DN), the cell viability was less in SiHa cells with cisplatin treatment, but these effects were not observed in HeLa cells. Similarly upon treatment with cisplatin SiHa cells had more activation of caspases compared to that seen in HeLa cells under conditions of NF kappaB inhibition by biological or chemical inhibitors. These results suggest that NF-kappaB may contribute to the resistance of human cervical cancer cells to cisplatin and highlight the potential use of combination therapy involving cisplatin and NF-kappaB inhibitors. PMID- 16044420 TI - Inhibition of epidermal growth factor-induced cell transformation and Akt activation by caffeine. AB - We found that caffeine significantly inhibited epidermal growth factor (EGF)- and 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced cell transformation in the JB6 mouse epidermal cell line. The tumor promoter-induced cell transformation was also blocked by treatment with an adenosine A1 receptor antagonist, 8 phenyltheophylline (8-PTH). Caffeine slightly attenuated activation of EGF induced activator protein 1 (AP-1) activation, which play important roles in cell transformation, but only at the highest concentration examined (1 mM). Interestingly, pretreatment with caffeine suppressed EGF-induced phosphorylation and activation of Akt and ribosomal p 70 S6 protein kinase (p 70 S 6 K), a target of Akt, without inhibiting phosphatidylinositol 3-kinase (PI 3 K) activation. The inhibition of Akt activation of caffeine was not a result of its adenosine receptor antagonism. Because Akt plays a key role in signal transduction pathways leading to cell proliferation and apoptosis, our results provide novel insight into possible mechanisms of the chemotherapeutic effect of caffeine. PMID- 16044421 TI - Joint properties of cast Fe-Pt magnetic alloy laser welded to gold alloys. AB - This study investigated the joint properties of a cast Fe-Pt magnetic alloy (Fe 36 at % Pt) laser welded to three gold alloys. The gold alloys used were ADA Type II and Type IV gold alloys, and an Ag-based (Ag-Au) gold alloy. Cast plates (0.5 x 3.0 x 10 mm) were prepared for each alloy. After the cast Fe-Pt plates were heat treated, they were butted against each of the three alloys and then laser welded with Nd:YAG laser at 200 V. Homogeneously welded specimens were also prepared for each alloy. Tensile testing was conducted at a crosshead speed of 1 mm/min. Failure load (N) and elongation (%) were recorded. After tensile testing, the fractured surfaces were examined with the use of SEM. The failure-load values of the group of alloys welded homogeneously were ranked in the order of: Ag-Au alloy > Type IV alloy > Type II alloy > Fe-Pt alloy. The Type IV alloy welded to Fe-Pt alloy had the highest failure-load value among the three alloys tested. The elongation results tended to follow a similar pattern. The results of this study indicated that Type IV gold alloy is a suitable alloy for metal frameworks to which cast Fe-Pt magnetic alloy is laser welded. PMID- 16044422 TI - Removal of surface by-products from sintered hydroxyapatite: effect of a chelation treatment on fibronectin adsorption and cell adhesion. AB - It was observed that fibronectin precipitates when deposited on hydroxyapatite (HA) ceramics. Fibronectin's known affinity for calcium and the composition of the ceramic itself suggested that calcium release could be the main cause of this aggregation effect. It was then decided to investigate the effect of a surface chelation treatment on fibronectin adsorption, and MG63 cell adhesion, onto porous ceramics of hydroxyapatite (HA), beta-tricalcium phosphate (beta-TCP), and HA/TCP biphasic material (BCP). Those ceramics were immersed in an EDTA solution and the effect of this treatment on the material composition was assayed. X-ray diffraction data showed the presence of alpha- and beta-TCP phases in HA and BCP materials, which were both completely removed by the chelation treatment in the case of HA. On BCP, alpha-TCP was removed and beta-TCP partially dissolved. The TCP material, which was pure beta-TCP, underwent a mass loss, but no change in composition was observed. Adhesion of MG63 cells was overall higher on the fibronectin-coated EDTA-treated HA material, but was especially enhanced on EDTA treated HA. Changes in surface morphologies, as compared with the use of scanning electron microscopy, did not seem to be related to the effects observed. The EDTA treatment proved to be a very efficient way of removing by-products of HA sintered materials, and thus enhancing the biocompatibility of the material. PMID- 16044423 TI - Histologic assessment of mesh fixation following laser-assisted tissue soldering in a lapine model. AB - OBJECTIVE: Wound histology and mesh bioincorporation following intraperitoneal fixation using laser-assisted soldering was evaluated. METHODS: 2.8-3.2 kg NZW rabbits underwent laparotomy. Controls had 2x2 cm segments of Mersilene stapled to peritoneum. Group 2 segments were affixed with 55% collagen solder onlay by fiber-coupled diode laser (1.43 +/- 10 micro, 2.5 W CW, 4 mm spot, 60 degrees C set temperature). Group 4 had Mersilene inlaid into melted solder. Group 3 had solder-embedded Vicryl mesh affixed. Animals were euthanized at 0, 2, 4, 6 weeks. Fixed sections were assessed for integrity, inflammation, and fibrosis using H & E, Masson's Trichrome and Evans Van Gieson staining. RESULTS: Histology demonstrated cell types, local mesh reaction, and progressive evidence of solder reabsorption mimicking normal healing and bioincorporation. Mersilene groups demonstrated normal arrangement of collagen-rich layers around mesh. CONCLUSION: Collagen-based tissue soldering permits normal wound healing and may mitigate use of staples. Further development of this strategy is warranted. PMID- 16044424 TI - Secondary metabolic effects in complex I deficiency. AB - The objective of this study was to investigate clinical, biochemical, and genetic features in 7 probands (a total of 11 patients) with nicotine-amide adenine dinucleotide (NADH) dehydrogenase (complex I) deficiency. We screened the mitochondrial DNA for mutations and found pathogenic mutations in complex I genes (mitochondrial NADH dehydrogenase subunit (MTND) genes) in three probands. The 10191T>C mutation in MTND3 and the 14487T>C mutation in MTND6 were present in two probands with Leigh's-like and Leigh's syndrome, respectively. Four siblings with a syndrome consisting of encephalomyopathy with hearing impairment, optic nerve atrophy, and cardiac involvement had the 11778G>A mutation in MTND4, previously associated with Leber hereditary optic neuropathy. These findings demonstrate that mutations in MTND genes are relatively frequent in patients with complex I deficiency. Biochemical measurements of respiratory chain function in muscle mitochondria showed that all patients had a moderate decrease of the mitochondrial adenosine triphosphate production rate. Interestingly, the complex I deficiency caused secondary metabolic alterations with decreased oxaloacetate induced inhibition of succinate dehydrogenase (complex II) and excretion of Krebs cycle intermediates in the urine. Our results thus suggest that altered regulation of metabolism may play an important role in the pathogenesis of complex I deficiency. PMID- 16044425 TI - Malignant transformation of high-grade anal intraepithelial neoplasia. AB - BACKGROUND: The natural history of anal intraepithelial neoplasia (AIN) is uncertain. This makes management problematic as treatment options to eradicate the condition carry morbidity. The authors report their 10-year experience with conservative management of this condition, highlighting the lessons learnt. METHODS: All patients were diagnosed with high-grade AIN (AIN III) between 1994 and 2003. Diagnosis was by full-thickness biopsy and histopathological examination. Excision of localized lesions was undertaken, and all patients underwent follow-up every 6 months. Prospective data were collected regarding recurrence, postoperative complications and progression to invasive carcinoma. RESULTS: Thirty-five patients were followed for a median of 63 (range 14-120) months. Excision of localized high-grade AIN was carried out in 28 patients with minimal morbidity. Six patients were systemically immunosuppressed at diagnosis, all of whom had multifocal perianal lesions. Three immunosuppressed patients developed invasive anal squamous carcinoma during follow-up. By contrast, no invasive carcinomas were identified among immunocompetent patients with either localized or multifocal perianal disease. CONCLUSION: AIN III appears to have a relatively low potential for malignant transformation in the immunocompetent patient. However, immunosuppressed patients are more likely to have extensive AIN III and a greater risk of malignant change. PMID- 16044426 TI - Sacral nerve stimulation reduces corticoanal excitability in patients with faecal incontinence. AB - BACKGROUND: Sacral nerve stimulation (SNS) can produce symptomatic relief in patients with faecal incontinence. Moreover, peripheral nerve stimulation has been shown to affect brain function. The aim of this study was to determine whether SNS might produce important changes in cortical activity linked to improved continence. METHODS: In an experimental study, ten women with intractable faecal incontinence (mean age 51.3 years) were serially mapped with transcranial magnetic stimulation before and immediately after 14 days of temporary SNS (15 Hz, pulse width 210 micros), and then 2 weeks later. Anal sphincter electromyographic responses were recorded to cortical stimulation of multiple points over a scalp grid covering the bilateral medial cortex. Continence scores, anorectal manometry and rectal sensitivity data were also collected. RESULTS: SNS improved global symptom scores in the majority of patients; mean(s.e.m.) continence scores fell from 16.9(1.6) to 10.6(1.8) (P = 0.042). Cortical mapping showed a consistent decrease in corticoanal representation and overall excitability immediately after SNS compared with baseline (mean(s.e.m.) 38,083(13,669) versus 42,507(13,297) microV s; P = 0.017), which was reversed 2 weeks after SNS wire removal. CONCLUSION: SNS produces symptom benefit in patients with faecal incontinence that is associated with a reversible reduction in corticoanal excitability. SNS therefore drives dynamic brain changes that may play a functional role in influencing anal continence. PMID- 16044427 TI - Ischaemic preconditioning improves microvascular perfusion and oxygenation following reperfusion injury of the intestine. AB - BACKGROUND: Ischaemia-reperfusion (IR) injury of the intestine occurs commonly during abdominal surgery. Ischaemic preconditioning (IPC) provides a way of protecting the organ from damage inflicted by IR. This study was designed to evaluate the beneficial effect of IPC, focusing on the intestinal microcirculation and oxygenation in intestinal IR injury. METHODS: Rats were allocated to three groups. Animals in the IR and IPC groups underwent 30 min of intestinal ischaemia followed by 2 h of reperfusion. In the IPC group this was preceded by 10 min of ischaemia and 10 min of reperfusion. Animals in the third group underwent laparotomy but no vascular occlusion. Intestinal microvascular perfusion, oxygenation and portal venous blood flow (PVF) were monitored continuously. At the end of the reperfusion period, blood samples were obtained for measurement of lactate dehydrogenase (LDH) and biopsies of ileum for histological evaluation. RESULTS: : IPC improved intestinal microvascular perfusion and tissue oxygenation significantly at the end of the reperfusion period (P < 0.001). PVF improved significantly in the IPC compared with the IR group (P = 0.005). The serum LDH concentration was significantly lower in the IPC than the IR group (mean(s.e.m.) 667.1(86.8) versus 1973.8(306.5) U/l; P < 0.001) Histological examination showed that ileal mucosa was significantly less injured in the IPC group. CONCLUSIONS: This study demonstrated that IPC improves intestinal microvascular perfusion and oxygenation. PMID- 16044428 TI - Biomechanical investigation of the effect of high hydrostatic pressure treatment on the mechanical properties of human bone. AB - Several methods are available for reconstruction of bone defects due to malignant tumors. To extracorporally devitalize resected tumor-bearing bone segments two methods, that is, extracorporal irradiation or autoclaving, are available up to now. However, both methods have substantial disadvantages like decrease of bone's mechanical strength. To develop an alternative method for tumor inactivation in skeletal segments, high hydrostatic pressure (HHP) was applied. Previous investigations have shown that human normal and tumor cell lines as well as tumor afflicted human bone specimens were irreversibly damaged at 350 MPa when subjected to HHP. This study was aimed to examine the alterations of biomechanical properties of human bone after exposure to HHP. Trabecular and cortical bone specimens were harvested from six pair of fresh-frozen human cadaveric femora. The bone specimens from one side were exposed to different pressure values of 300 or 600 MPa over 10 min. Bone samples from the contralateral sites were used as untreated controls. Biomechanical properties were investigated by a quasi-static compression test for trabecular specimens and by a quasi-static four-point bending test for cortical specimens, respectively. Biomechanical properties of the cortical and trabecular bone did not decrease after exposure to 300 MPa regarding the testing parameters Young's modulus and ultimate strength (200.7 +/- 38.7 MPa for HHP treated cortical bone versus 186.5 +/- 34.3 MPa for the untreated control group). After pressure treatment at 600 MPa Young's modulus and ultimate strength respectively remained almost unchanged in trabecular bone and were reduced about 15% in cortical bone (p < 0.001 and p =0.002, respectively). We anticipate that in orthopedic surgery HHP can serve as a novel, promising methodical approach for tumor cell inactivation, which occurs at pressure levels of about 300 MPa. Thereby immediate reimplantation of treated bone segments by preservation of the essential biomechanical properties of bone could become possible. Even after HHP treatment at 600 MPa the strength of bone only decreases up to 15%. PMID- 16044429 TI - The neuroprotective KDI domain of gamma 1-laminin is a universal and potent inhibitor of ionotropic glutamate receptors. AB - Previous work from this laboratory indicates that the KDI (Lys-Asp-Ile) domain of gamma 1-laminin promotes functional regeneration of adult rat spinal cord injuries and protects adult rat hippocampal neurons against massive neuronal death induced by intracerebral injection of the glutamate analogue kainic acid. In the present study, we used patch clamp recordings on cultured human embryonic neocortical neurons and HEK 293 cells expressing recombinant glutamate receptor subunits to study a putative interaction of the KDI with the glutamate system. We show that the KDI domain of gamma 1-laminin is a universal and potent inhibitor of AMPA, kainate, and NMDA subclasses of glutamate receptors, with a noncompetitive action on the AMPA receptor channel activity. Glutamate neurotoxicity plays a key role in both CNS trauma and neurodegenerative disorders, so this unexpected, novel function of the gamma 1-laminin-derived tripeptide may prove clinically valuable in treatment of CNS trauma and/or disease. PMID- 16044430 TI - Biomimetic porous scaffolds with high elasticity made from mineralized collagen- an animal study. AB - Histological investigations of a new hydroxyapatite-collagen composite material were carried out to evaluate its possible suitability as a bone substitute. The three-dimensional scaffolds made from biomimetically mineralized collagen exhibit an interconnecting pore structure and elastic mechanical properties. They were implanted into the subcutaneous tissue and bone defects made in the femur of rats and harvested with the surrounding tissue at 1, 2, 4, 8, and 12 weeks after surgery. The materials implanted in the subcutaneous tissue were covered by fibrous connective tissue with a slight inflammatory response, and many foreign body giant cells were observed on the surface of the scaffolds. Most of the material implanted in the subcutaneous tissue was resorbed at 8 weeks by phagocytosis. In the bone defects, new bone formation was observed on the surface of the material at 1 week. New bone increased with time, and osteoclasts were seen on the surface of the scaffolds at 2 weeks. Resorption and replacement by new bone of many parts of the materials implanted in the femur were observed by 12 weeks. These responses occurred faster than those of other hydroxyapatite collagen composites. The results suggested that the new biomimetically mineralized collagen scaffolds were suitable as an implant material for bone tissue reconstruction. PMID- 16044431 TI - Collagen-immobilized poly(vinyl alcohol) as an artificial cornea scaffold that supports a stratified corneal epithelium. AB - The cornea is a transparent tissue of the eye, which is responsible for the refraction of incoming light. Both biological corneal equivalents and synthetic keratoprostheses have been developed to replace donor tissue as a means to restore vision. However, both designs have drawbacks in terms of stability and biocompatibility. Clinically available synthetic devices do not support an intact epithelium, which poses a risk of microbial infection or protrusion of the prosthesis. In the present study, type I collagen was immobilized onto poly(vinyl alcohol) (PVA-COL) as a possible artificial cornea scaffold that can sustain a functional corneal epithelium. Human and rabbit corneal epithelial cells were air lift cultured with 3T3 feeder fibroblasts to form a stratified epithelial layer on PVA-COL. The epithelial sheet expressed keratin 3/12 differentiation markers, the tight junction protein occludin, and had characteristic microvilli structures on transmission electron microscopy. Functionally, the stratified epithelium contained normal glycogen levels, and an apical tight-junction network was observed to exclude the diffusion of horseradish peroxidase. Furthermore, the epithelium-PVA-COL composite was suturable in the rabbit cornea, suggesting the possibility of using PVA-COL as a biocompatible material for keratoprosthesis. PMID- 16044432 TI - Structural features and mechanical properties of in situ-bonded meshes of segmented polyurethane electrospun from mixed solvents. AB - The relationships between the structural features and mechanical properties of electrospun segmented polyurethane (SPU) meshes and electrospinning parameters such as formulation (e.g., polymer concentration and solvent mixing ratio) and operation parameters (e.g., applied voltage, air gap, and flow rate) were studied with the use of a mixed-solvent system of tetrahydrofuran (THF) and N,N dimethylacrylamide (DMF). After the relationships between the structure of electrospun SPU and the operation parameters under a fixed SPU concentration of single THF solution were established, SPU was electrospun from the mixed solvent of THF and DMF with different mixing ratios [DMF content: 5, 10, and 30% (v/v)]. Scanning electron microscopy showed that an increase in DMF ratio significantly enhances the degree of bonding between SPU fibers at contact sites and decreases the diameter of fibers formed. The porosimetric characterization showed the following: (1) The porosity of the electrospun SPU meshes decreased with an increase of DMF ratio. (2) The pore size distribution exhibited three representative peaks of different void sizes (i.e., approximately 5, 20, and 70 microm). (3) The proportion of the 20-microm void markedly decreased with an increase in DMF ratio. A tensile test on the meshes showed that an increase in DMF ratio induces an increase in elasticity of the mesh. Such a regulation of the structural features and mechanical properties of electrospun SPU meshes using a mixed-solvent system with low- and high-boiling-point solvents may be useful in the engineering of SPU-fiber based matrices or scaffolds. PMID- 16044433 TI - Dual-level direct dynamics studies on the reaction Cl + CHBr(2)Cl. AB - Theoretical investigations are carried out on the multichannel reaction CHBr(2)Cl + Cl by means of direct dynamics methods. The minimum energy path (MEP) is obtained at the BH&H-LYP/6-311G(d,p) level, and energetic information is further refined at the CCSD(T)/6-311+G(2df,2p) (single-point) level. The rate constants for three reaction channels, H-abstraction, Br-abstraction, and Cl-abstraction, are calculated by using the improved canonical variational transition state theory (ICVT) incorporating with the small-curvature tunneling (SCT) correction. The theoretical overall rate constants are in good agreement with the available experimental data and are found to be k=2.58 x 10(-15) T(1.18) exp(-861.17/T) cm(3)molecule(-1)s(-1) over the temperature range 200--2400 K. For the title reaction, H-abstraction reaction channel is the major channel at the lower temperatures, while as the temperature increases, the contribution of Br abstraction reaction channel should be taken into account. At 2180 K, the rate constants of these two pathways are equal. Cl-abstraction reaction channel is minor channel over the whole temperature region. PMID- 16044434 TI - Deterministic global optimization of molecular structures using interval analysis. AB - The search for the global minimum of a molecular potential energy surface is a challenging problem. The molecular structure corresponding to the global minimum is of particular importance because it usually dictates both the physical and chemical properties of the molecule. The existence of an extremely large number of local minima, the number of which may increase exponentially with the size of the molecule, makes this global minimization problem extremely difficult. A new strategy is described here for solving such global minimization problems deterministically. The methodology is based on interval analysis, and provides a mathematical and computational guarantee that the molecular structure with the global minimum potential energy will be found. The technique is demonstrated using two sets of example problems. The first set involves a relatively simple potential model, and problems with up to 40 atoms. The second set involves a more realistic potential energy function, representative of those in current use, and problems with up to 11 atoms. PMID- 16044435 TI - POEMS syndrome: failure of newly suggested diagnostic criteria to anticipate the development of the syndrome. AB - POEMS syndrome is a unique clinical entity. Although it's a diagnosis of exclusion, it was previously described by the presence of several typical characteristics as paraproteinemia, polyneuropathy, organomegaly, endocrinopathy, and skin changes. Recently, new criteria were proposed, and the presence of two major and one minor criterion was claimed to suffice for a diagnosis. Both methods considered other important characteristics germane to the syndrome unessential for diagnosis. Retrospective evaluation of patients with lymphoproliferative disease was carried out to reveal the presence of the syndrome according to these different methods. Patients' clinical progression during follow-up will be used to validate the criteria's sensitivity and specificity. Six hundred twenty-nine consecutive files of patients with paraproteinemia who were followed-up at a tertiary medical center were reviewed. Of 12 patients who fulfilled the new criteria for diagnosis of POEMS, 3 remain stable during long-term follow-up and only 5 finally developed the full-blown syndrome. Four patients developed other diseases that accounted for their clinical findings. Patients presenting with neuronal vasculitic changes on biopsy, kappa light-chain monoclonal gammopathy, and cryoglobulinemia were unlikely to develop POEMS syndrome, even though they fulfilled the newly suggested criteria. Although they are not in the criteria, sclerotic bone lesions were found only in patients who eventually developed the full syndrome. The diagnosis of POEMS syndrome according to the newly suggested criteria should not be definitive in the presence of atypical clinical features of the syndrome. PMID- 16044436 TI - Clinical importance of bone marrow monocytic nodules in patients with myelodysplasia: retrospective analysis of 21 cases. AB - Bone marrow monocytic nodules (MNs) can occur in various myeloid disorders. This retrospective review identified 21 patients with myelodysplasia who had unusual and distinct MNs. Eight patients had chronic myelomonocytic leukemia (CMML); 4 had acute myeloid leukemia (AML); and 9 had myelodysplastic/myeloproliferative diseases. In each case, the cells forming MNs expressed strong CD68. MNs appeared to persist even after aggressive chemotherapy, including conventional chemotherapy for 2 AML patients and high-dose chemotherapy preceding allogeneic bone marrow transplantation for 1 CMML patient. Thirteen of 21 patients (62%) died, and acute leukemic transformation was the main cause of death in 3 of 8 patients with CMML. The median survival of the 20 patients with appropriate follow-up was 9.8 months. Our findings demonstrate that MNs are associated with CMML, AML, myelodysplastic syndromes, and myeloproliferative diseases and suggest that MNs are resistant to intensive chemotherapy and patients with bone marrow MNs have a poor prognosis. PMID- 16044437 TI - Large granular lymphocytic proliferation-associated cyclic thrombocytopenia. AB - Cyclic thrombocytopenia is a rare condition characterized by regular fluctuations in the platelet count, resulting in bleeding at the time of platelet count nadir. We evaluated a male patient whose platelet count cycled between <10x10(9)/L and a maximum of >1300x10(9)/L over a median of every 42 days (range, 28-57 days). Antiplatelet antibodies were present at highest titer just prior to platelet nadirs. A pathologic expansion of CD3+CD57+ large granular lymphocytes (LGLs) along with a clonal rearrangement of the T-cell receptor (TCR) gamma gene in circulating T cells was detected. LGLs decreased in response to treatment with cyclosporine-A (CsA), but the cycling of the platelet count continued. This is the first report of cyclic thrombocytopenia associated with a T-LGL lymphoproliferative disorder. PMID- 16044438 TI - Failure of rituximab in human immunodeficiency virus-associated multicentric Castleman disease. AB - Two human immunodeficiency virus-infected patients who needed etoposide therapy to control exacerbations of Castleman disease received four infusions of rituximab. Clinical and virologic relapses with increased blood human herpesvirus 8 DNA levels occurred in both patients 4 and 24 weeks later and were associated with a worsening of Kaposi sarcoma. PMID- 16044439 TI - Diagnosis of deep vein thrombosis by plasma-soluble fibrin or D-dimer. AB - The present study was designed to determine the cutoff values of D-dimer and soluble fibrin (SF) for the diagnosis of deep venous thrombosis (DVT) and pulmonary embolism (PE) in Japanese patients. Plasma levels of these molecules were measured in 243 patients suspected of having DVT and 100 healthy volunteers (controls). Out of 243 patients, 20 patients were diagnosed with DVT. In the control group, plasma levels of D-dimer and SF did not show normal distribution, and the 95% confidence intervals (CI) of D-dimer and SF were 2.45 microg/mL and 6.92 microg/mL, respectively. Plasma levels of D-dimer and SF of patients with DVT were significantly higher than of those without DVT. In patients with DVT, the minimum values of D-dimer and SF were 1.71 and 1.44 microg/mL, respectively. When the cutoff value was set at the average+1 SD of those of the control (D dimer, about 1.8 microg/mL; SF, about 1.4 microg/mL), 1 and 0 patient with DVT was overlooked, respectively. The sensitivity and specificity of D-dimer and SF for DVT were 95% and 100%, and 61.9% and 53.8%, respectively. When the cutoff value was set at 95% CI of the control (D-dimer, 2.5 microg/mL; SF, 6.9 microg/mL), 2 and 9 patients with DVT were overlooked, respectively. The sensitivity and specificity of D-dimer and SF were 90% and 50%, and 77.6% and 88.3%, respectively. When the cutoff values set at 2.5 microg/mL of D-dimer or 6.9 microg/mL of SF, 1 DVT patient was overlooked, with sensitivity and specificity of 95% and 69.5%. Our data suggest that both D-dimer and SF are useful markers for the diagnosis of DVT and that measurement of both D-dimer and SF increases the sensitivity and specificity for the diagnosis of DVT/PE. PMID- 16044440 TI - Meningeal hematopoiesis following radiation myelitis in a hematopoietic stem-cell transplant recipient. AB - Extramedullary meningeal hematopoiesis (EMH) represents an uncommon finding after stem-cell transplantation. We describe the case of an allogeneic bone marrow transplantation (BMT) recipient who developed EMH 1 month after radiation myelitis had been diagnosed. A 39-year-old man with multiple myeloma underwent matched unrelated BMT following a myeloablative conditioning regimen of cyclophosphamide and total-body irradiation (200 cGyx6). This was followed by delivery of 40 Gy of involved-field radiation to an extramedullary plasmacytoma compressing the spinal cord. Although transplantation went extremely well, the patient developed radiation myelitis 7 months after transplantation, and EMH ensued 1 month later. Because the patient was not in a disease state known to cause EMH, it is tempting to speculate that radiation-related neural injuries might cause donor cells to migrate to the central nervous system. PMID- 16044441 TI - Donor cell derived acute myeloid leukemia after allogeneic cord blood transplantation in a patient with adult T-cell lymphoma. AB - We report a patient with adult T-cell lymphoma who developed acute myeloid leukemia (AML) after allogeneic cord blood transplantation (CBT). Fluorescence in situ hybridization (FISH) studies and molecular analysis using short tandem repeat (STR) sequences proved the AML to be of donor origin. Although 25 cases of donor cell leukemia (DCL) occurring after allogeneic bone marrow transplantation have previously been reported, there have been no reports of DCL after CBT. This case is the first-reported DCL patient after CBT. PMID- 16044442 TI - Perioperative management of a patient with May-Hegglin anomaly requiring craniotomy. AB - May-Hegglin anomaly (MHA) is a rare type of autosomal dominant platelet disorder associated with mutations in the gene encoding nonmuscle myosin heavy chain 9 (MYH9). It is characterized by the presence of large platelets, leukocyte inclusions, and thrombocytopenia. The bleeding tendency is usually mild, but severe hemorrhages have been reported. This is the first reported case of a patient with MHA who underwent craniotomy for intractable seizure disorder of temporal lobe origin. Patients who have thrombocytopenia have a higher likelihood of developing intraoperative or postoperative intracranial hematoma and bleeding complications. The patient was administered desmopressin (DDAVP) prior to the neurosurgical procedure and had no complications. With this approach, the use of platelet concentrates could be avoided. We discuss the role of DDAVP in MHA and related platelet disorders and review the current literature. PMID- 16044443 TI - Intraneutrophilic malaria rings: a rare finding. PMID- 16044444 TI - Definition of organ involvement and treatment response in immunoglobulin light chain amyloidosis (AL): a consensus opinion from the 10th International Symposium on Amyloid and Amyloidosis, Tours, France, 18-22 April 2004. AB - We undertook this study to develop uniformly accepted criteria for the definition of organ involvement and response for patients on treatment protocols for immunoglobulin light-chain amyloidosis (AL). A consensus panel was convened comprising 13 specialists actively involved in the treatment of patients with amyloidosis. Institutional criteria were submitted from each, and a consensus was developed defining each organ involved and the criteria for response. Specific criteria have been developed with agreed on definitions of organ and hematologic response as a result of discussions at the 10th International Symposium on Amyloid and Amyloidosis held in Tours, France, April 2004. These criteria now form the working definition of involvement and response for the purposes of future data collection and reporting. We report criteria that centers can now use to define organ involvement and uniform response criteria for reporting outcomes in patients with light-chain AL. PMID- 16044445 TI - Clonal T cells of pure red-cell aplasia. AB - This study detected clonal T cells in patients with acquired pure red-cell aplasia (PRCA) by Southern blotting and polymerase chain reaction (PCR). Twenty nine adult patients with acquired PRCA were enrolled in this study. Seventeen patients had primary acquired PRCA, while 12 patients had the secondary form. Twenty-two of 29 (76%) patients demonstrated TCR rearrangement by at least one method. We divided the patients into three groups depending on T-cell clonality. The CD4/8 ratio of patients who were positive on Southern blotting was significantly lower than that of other groups. Except for the CD4/8 ratio, other laboratory findings did not significantly differ among the three groups. The CD4/8 ratio should be a useful surrogate marker to detect T-cell clonality. PMID- 16044446 TI - Erythrophagocytosis by dysplastic neutrophils in chronic myelomonocytic leukemia and subsequent transformation to acute myeloid leukemia. AB - Erythrophagocytosis by neutrophils is a rare phenomenon in myeloid malignancies, and its clinicopathologic significance is not fully understood. We report a unique case of erythrophagocytosis by dysplastic neutrophils in chronic myelomonocytic leukemia (CMML) and subsequent transformation to acute myeloid leukemia (AML). Review of multiple marrow samples, both pretreatment and post treatment, demonstrated a correlation between percentage of dysplastic neutrophils and degree of erythrophagocytosis. Erythrophagocytosis was observed only in dysplastic forms of neutrophils. Post-transplant marrows with engraftment of donor cells showed no neutrophilic dysplasia or erythrophagocytosis. Possible mechanisms of neutrophilic erythrophagocytosis in myeloid malignancies are discussed. PMID- 16044447 TI - Successful treatment of intestinal hemorrhage in a Jehovah's Witness patient. PMID- 16044448 TI - Bleeding complication under Gingko biloba medication. PMID- 16044449 TI - hTERT mRNA levels by real-time RT-PCR in acute myelogenous leukemia. AB - The purpose of this study was to investigate whether levels of hTERT mRNA, as determined by real-time RT-PCR, are associated with prognosis and clinical course in AML patients. Fifty-four bone marrow specimens from 21 patients diagnosed with de-novo AML were included. The level of hTERT mRNA was measured with the Telo TAGGG hTERT Quantification Kit (Roche Diagnostics, Mannheim, Germany), using a LightCycler Instrument (Roche Diagnostics). The level of hTERT mRNA was determined as the relative ratio (RR), which was calculated by dividing the level of hTERT mRNA by the level of the porphobilinogen deaminase (PBGD) housekeeping gene in the same samples [1,000x(hTERT/PBGD)]. The expression rates of hTERT mRNA were significantly higher at diagnosis (73%) and during relapse (80%) than during remission (27%) (P<0.05). The median RR for diagnosis or relapse was significantly higher than that for patients in remission (P<0.05). hTERT mRNA expression was not correlated with CD34 expression, blast counts, white blood cell counts, or chromosomal abnormality (P>0.05). Two patients who showed hTERT mRNA expression during remission (RR 3.14 and 7.15, respectively) relapsed after 1 month. Among seven patients with high hTERT mRNA levels (RR>9.51), 4 failed to achieve complete remission (CR), whereas 4 of 5 patients without hTERT mRNA expression at diagnosis or during relapse achieved CR (P>0.05). Patients showing a trend of increasing hTERT mRNA levels failed to reach a second CR after relapse, while those with a trend toward decreasing hTERT mRNA did achieve CR. Among eight samples showing hTERT mRNA expression in remission (RR>0), 5 were obtained from patients who had received GCSF within 14 days. The expression rate and level of hTERT mRNA during remission were significantly higher in patients who had previously received GSCF (56%, RR=0.15) than in other patients (15%, RR=0) (P<0.05). Serial and quantitative analysis of hTERT mRNA may be a useful marker for prediction of prognosis and monitoring in AML patients. PMID- 16044450 TI - Passive donor-to-recipient transfer of antiphospholipid syndrome following allogeneic stem-cell transplantation. AB - Autoantibody production following allogeneic stem-cell transplantation is common and is often ascribed to the immune dysregulation associated with graft-versus host disease. Recent data suggests that donor-memory B cells can be reactivated on exposure to antigen and result in antibody production in the recipient identical to that seen in the donor. Here we describe the production of autoantibodies in a recipient of bone marrow from a donor with systemic lupus erythematosus and antiphospholipid syndrome. Autoantibody appearance was precipitated by the onset of graft-versus-host disease, was identical to that of the donor, and ultimately lead to cerebrovascular thrombosis, which was successfully treated with antiplatelet and anticoagulant therapy. PMID- 16044451 TI - Comment on hemostatic markers and the sepsis-related organ failure assessment score in patients with disseminated intravascular coagulation in an intensive care unit by Okabayashi et al. PMID- 16044453 TI - PRAME mRNA levels in cases with acute leukemia: clinical importance and future prospects. AB - The PRAME (preferentially expressed antigen of melanoma) gene has been shown to be expressed in high levels in some solid tumors and hemopoietic neoplasias but not or only weakly expressed in normal tissues. It encodes an antigen recognized by autologous cytolytic T lymphocytes. PRAME is a good candidate for tumor immunotherapy and is a useful marker gene for detection of minimal residual disease (MRD). In this study, PRAME mRNA using real-time RT-PCR was studied in 74 adult cases with acute leukemia-68 had de-novo acute leukemia, 3 had chronic myeloid leukemia-blastic crisis (CML-BC), and 3 had myelodysplastic/myeloproliferative syndrome-blastic transformation (MDS/MPD-BT) and the results were compared with 30 age-matched healthy volunteers. Nineteen of 74 cases with leukemia expressed PRAME, while only 2 controls showed weak expression. The prevalence of PRAME expression in AML and ALL cases was 30% and 17%, respectively. We did not find any important correlation between PRAME expression and clinical characteristics, such as age, sex, organomegaly/lymphadenopathy, Hb, WBC count, platelet count, LDH level, alkaline phosphatase, albumin, cell-surface antigens, response to therapy, or progression free and overall survival. PRAME was monitored in 15 cases during remission and/or relapse. There was a good correlation between PRAME mRNA and hematological remission and/or relapse. Interestingly, PRAME was very high in one case with AML but was not found 3 months after allogeneic transplantation. PRAME mRNA is observed in about one-third of AML cases; it may be a useful marker to detect MRD, and it may also be a good predictor for the timing of donor lymphocyte infusions (DLI) in the post-transplant period in cases of molecular relapse. PMID- 16044454 TI - Mutations in the MCFD2 gene and a novel mutation in the LMAN1 gene in Indian families with combined deficiency of factor V and VIII. AB - Combined deficiency of factors V (FV) and factor VIII (FVIII) (F5F8D) is an autosomal recessive bleeding disorder caused by simultaneous moderate-to-mild decrease of both clotting proteins. Mutations in two components of the ER-Golgi intermediate compartment (ERGIC-53), i.e., lectin mannose binding protein (LMAN1) and multiple coagulation factor deficiency 2 (MCFD2), have been found to be responsible for this dual deficiency in most of the cases reported in literature. Three Indian families with F5F8D were analyzed for the presence of mutations in their LMAN1 and MCFD2 genes. One of the three families showed the presence of a G to A substitution in exon 2 of the MCFD2 gene, whereas another family showed a nonsense mutation, i.e., G to T substitution, in exon 2 of the LMAN1 gene, the latter being a novel mutation not previously reported. The third family did not show mutations in either of the two genes, suggesting that a significant subset of F5F8D cases may be due to additional genes resulting in a similar phenotype. PMID- 16044455 TI - Immunological study of complex formation between soluble transferrin receptor and transferrin. AB - Transferrin receptor (TfR) is a dimeric transmembrane protein that provides iron transport from plasma to cells by binding and internalization of iron-loaded transferrin (Tf). Soluble transferrin receptor (sTfR) is an extracellular part of the TfR molecule that is truncated from the cell surface and released into the blood stream. Using monoclonal antibodies (HyTest Ltd., Turku, Finland), immunofluorescent methods for sTfR and sTfR-Tf complex determination were developed. Soluble TfR was isolated from human plasma, and complex formation between sTfR and Tf was studied by stepwise complex construction and by FPLC gel filtration. It was found that sTfR could bind two Tf molecules step by step when the sTfR-Tf complex is constructed in the plate wells. FPLC gel filtration of sTfR-Tf mixtures and analysis of sTfR and sTfR-Tf immunological activities in collected fractions showed that sTfR can form different complexes with TF depending upon the ratios between them: a 291-kDa compound is assumed to be a 2:1 sTfR/Tf complex, and a 345-kDa compound is assumed to be a 2:2 sTfR/Tf complex. Isolated sTfR eluted as a 237-kDa protein. FPLC gel filtration of serum revealed that all sTfR in serum is bound to Tf in a 2:2 complex, and no isolated sTfR can be found in serum. This raises the question as to the nature of the bonds that hold two molecules of sTfR together to form a dimer. PMID- 16044456 TI - Therapy-related acute myeloid leukemia after single-agent treatment with fludarabine for chronic lymphocytic leukemia. AB - A 70-year-old man with B-cell chronic lymphocytic leukemia (CLL) received single agent treatment with the purine analogue fludarabine, which led to complete remission. After 8 years, he presented with pancytopenia. Marrow examination showed acute myeloid leukemia (AML) with trilineage myelodysplasia (MDS). Cytogenetic analysis showed an unbalanced der(1;7)(p10;q10) that resulted effectively in deletion 7q; confirming the diagnosis of therapy-related AML (t AML). No residual CLL was present. Together with previous reports of secondary cancers after fludarabine treatment and the association of monosomy 7/7q- with another purine analogue azathioprine, results suggest that t-AML might develop after fludarabine therapy. PMID- 16044457 TI - Interphase FISH screening for the LCR-mediated common rearrangement of isochromosome 17q in primary myelofibrosis. AB - Non-allelic homologous recombination (NAHR) between low-copy repeats (LCRs) has been implicated recently in somatic rearrangements including isochromosome i(17q), which is associated with hematologic malignancies as well as solid tumors. In hematological malignancies, the most common i(17q) breakpoint results from LCR-mediated NAHR, which creates a dicentric chromosome, idic(17)(p11.2). We report an elderly patient who presented with primary myelofibrosis (MF) with myeloid metaplasia (MMM), associated with idic(17)(p11.2) as the sole chromosomal abnormality, making this the first idic(17)(p11.2) myeloproliferative case reported in which the breakpoints are mapped to the breakpoint cluster region in proximal 17p. The rearrangement breakpoint maps to the previously defined LCR cluster, further suggesting that the genomic architecture of proximal 17p may be responsible for the formation of the majority of i(17q) cases. We describe our development of a rapid screening test using interphase FISH to detect idic(17)(p11.2), discuss the potential prognostic value of this molecular diagnostic test, and examine the relevance of LCR-mediated NAHR to common rearrangements in neoplasms. PMID- 16044458 TI - Nucleotide -88 (C-T) promoter mutation is a common beta-thalassemia mutation in the Jat Sikhs of Punjab, India. AB - A study of beta-gene mutations in Jat Sikhs, a subcaste of Punjabis, revealed a very high prevalence (46%: 41/88) of the mild beta++ promoter region mutation -88 (C-T). Sixteen individuals presenting in homozygous form were clinically mild. Un transfused patients had characteristic hematological findings: high Hb F (38.1 68.6%, mean 47.4%), high Hb A2 (5.7-9.8%, mean 6.88%), and the rest had adult hemoglobin. The 19 subjects with compound heterozygosity for -88 (C-T) and another beta-gene mutation presented both as thalassemia intermedia (four cases) and as thalassemia major (15 cases). One of the four patients with the milder phenotype had a second mild mutation, CAP+1 (beta++). In the other three cases with the milder phenotype, the second mutation was associated with the presence of the XmnI Ggamma polymorphism. Notably, the XmnI Ggamma was negative in all the -88 (C-T) alleles. None of the patients had associated alpha-thalassemia even in the thalassemia intermedia group. Haplotype analysis of the -88 (C-T) homozygous cases showed a single haplotype (+ - - - - + -) in all but two individuals. This haplotype is distinct from those described in the Africans with homozygous -88 (C T), suggesting that the mutation in our population occurred independently. PMID- 16044459 TI - The influence of ultrasound on the release of gentamicin from antibiotic-loaded acrylic beads and bone cements. AB - Gentamicin-loaded acrylic beads are loosely placed in infected bone cavities, whereas gentamicin-loaded acrylic bone cement is used as a mechanical filler in bone to anchor prosthetic components. Both drug delivery systems are used to decrease infection rates by gentamicin release. The objective of this study is to investigate the effects of pulsed ultrasound on gentamicin release from both materials. Gentamicin release from gentamicin-loaded beads (Septopal) and from three commercially-available brands of gentamicin-loaded bone cement (CMW 1, Palacos R-G, and Palamed G) was measured after 18 h of exposure in PBS to an ultrasonic field of 46.5 kHz in a 1:3 duty cycle with an average acoustic intensity of 167 mW/cm(2). Samples not exposed to ultrasound were used as controls. Pulsed ultrasound significantly enhanced gentamicin release from gentamicin-loaded beads, whereas gentamicin release from the gentamicin-loaded bone cements was not significantly enhanced. Mercury intrusion porosimetry revealed an increased distribution of pores between 0.1 and 0.01 microm in beads after gentamicin release, while in bone cements no increase in the number of pores was found. Increased gentamicin release in beads due to ultrasound may be explained by micro-streaming in a porous structure, whereas the absence of changes in pore structure after gentamicin release in bone cement is concurrent with the lack of an enhanced release of the antibiotic by ultrasound. As an effective treatment of infections requires high local concentrations of antibiotic, increased gentamicin release due to ultrasound may be of clinical significance, especially since ultrasound has been demonstrated to increase bacterial killing by antibiotics. PMID- 16044460 TI - Interactions of macromolecules with salt ions: an electrostatic theory for the Hofmeister effect. AB - Salting-out of proteins was discovered in the nineteenth century and is widely used for protein separation and crystallization. It is generally believed that salting-out occurs because at high concentrations salts and the protein compete for solvation water. Debye and Kirkwood suggested ideas for explaining salting out (Debeye and MacAulay, Physik Z; 1925;131:22-29; Kirkwood, In: Proteins, amino acids and peptides as ions and dipolar ions. New York: Reinhold; 1943. p 586 622). However, a quantitative theory has not been developed, and such a theory is presented here. It is built on Kirkwood's idea that a salt ion has a repulsive interaction with an image charge inside a low dielectric cavity. Explicit treatment is given for the effect of other salt ions on the interaction between a salt ion and its image charge. When combined with the Debye-Huckel effect of salts on the solvation energy of protein charges (i.e., salting-in), the characteristic curve of protein solubility versus salt concentration is obtained. The theory yields a direct link between the salting-out effect and surface tension and is able to provide rationalizations for the effects of salt on the folding stability of several proteins. PMID- 16044461 TI - The folding landscape of an alpha-lytic protease variant reveals the role of a conserved beta-hairpin in the development of kinetic stability. AB - Most secreted bacterial proteases, including alpha-lytic protease (alphaLP), are synthesized with covalently attached pro regions necessary for their folding. The alphaLP folding landscape revealed that its pro region, a potent folding catalyst, is required to circumvent an extremely large folding free energy of activation that appears to be a consequence of its unique unfolding transition. Remarkably, the alphaLP native state is thermodynamically unstable; a large unfolding free energy barrier is solely responsible for the persistence of its native state. Although alphaLP folding is well characterized, the structural origins of its remarkable folding mechanism remain unclear. A conserved beta hairpin in the C-terminal domain was identified as a structural element whose formation and positioning may contribute to the large folding free energy barrier. In this article, we characterize the folding of an alphaLP variant with a more favorable beta-hairpin turn conformation (alphaLP(beta-turn)). Indeed, alphaLP(beta-turn) pro region-catalyzed folding is faster than that for alphaLP. However, instead of accelerating spontaneous folding, alphaLP(beta-turn) actually unfolds more slowly than alphaLP. Our data support a model where the beta-hairpin is formed early, but its packing with a loop in the N-terminal domain happens late in the folding reaction. This tight packing at the domain interface enhances the kinetic stability of alphaLP(beta-turn), to nearly the same degree as the change between alphaLP and a faster folding homolog. However, alphaLP(beta-turn) has impaired proteolytic activity that negates the beneficial folding properties of this variant. This study demonstrates the evolutionary limitations imposed by the simultaneous optimization of folding and functional properties. PMID- 16044462 TI - BAliBASE 3.0: latest developments of the multiple sequence alignment benchmark. AB - Multiple sequence alignment is one of the cornerstones of modern molecular biology. It is used to identify conserved motifs, to determine protein domains, in 2D/3D structure prediction by homology and in evolutionary studies. Recently, high-throughput technologies such as genome sequencing and structural proteomics have lead to an explosion in the amount of sequence and structure information available. In response, several new multiple alignment methods have been developed that improve both the efficiency and the quality of protein alignments. Consequently, the benchmarks used to evaluate and compare these methods must also evolve. We present here the latest release of the most widely used multiple alignment benchmark, BAliBASE, which provides high quality, manually refined, reference alignments based on 3D structural superpositions. Version 3.0 of BAliBASE includes new, more challenging test cases, representing the real problems encountered when aligning large sets of complex sequences. Using a novel, semiautomatic update protocol, the number of protein families in the benchmark has been increased and representative test cases are now available that cover most of the protein fold space. The total number of proteins in BAliBASE has also been significantly increased from 1444 to 6255 sequences. In addition, full-length sequences are now provided for all test cases, which represent difficult cases for both global and local alignment programs. Finally, the BAliBASE Web site (http://www-bio3d-igbmc.u-strasbg.fr/balibase) has been completely redesigned to provide a more user-friendly, interactive interface for the visualization of the BAliBASE reference alignments and the associated annotations. PMID- 16044463 TI - Different methylation characteristics of protein arginine methyltransferase 1 and 3 toward the Ewing Sarcoma protein and a peptide. AB - The multifunctional Ewing Sarcoma (EWS) protein, a member of a large family of RNA-binding proteins, is extensively asymmetrically dimethylated at arginine residues within RGG consensus sequences. Using recombinant proteins we examined whether type I protein arginine methyltransferase (PRMT)1 or 3 is responsible for asymmetric dimethylations of the EWS protein. After in vitro methylation of the EWS protein by GST-PRMT1, we identified 27 dimethylated arginine residues out of 30 potential methylation sites by mass spectrometry-based techniques (MALDI-TOF MS and MS/MS). Thus, PRMT1 recognizes most if not all methylation sites of the EWS protein. With GST-PRMT3, however, only nine dimethylated arginines, located mainly in the C-terminal region of EWS protein, could be assigned, indicating that structural determinants prevent complete methylation. In contrary to previous reports this study also revealed that trypsin is able to cleave after methylated arginines. Pull-down experiments showed that endogenous EWS protein binds efficiently to GST-PRMT1 but less to GST-PRMT3, which is in accordance to the in vitro methylation results. Furthermore, methylation of a peptide containing different methylation sites revealed differences in the site selectivity as well as in the kinetic properties of GST-PRMT1 and GST-PRMT3. Kinetic differences due to an inhibition effect of the methylation inhibitor S adenosyl-L-homocysteine could be excluded by determining the corresponding K(i) values of the two enzymes and the K(d) values for the methyl donor S-adenosyl-L methionine. The study demonstrates the strength of MS-based methods for a qualitative and quantitative analysis of enzymic arginine methylation, a posttranslational modification that becomes more and more the object of investigations. PMID- 16044464 TI - South Amerindian craniofacial morphology: diversity and implications for Amerindian evolution. AB - The most compelling models concerning the peopling of the Americas consider that modern Amerindians share a common biological pattern, showing affinities with populations of the Asian Northeast. The aim of the present study was to assess the degree of variation of craniofacial morphology of South American Amerindians in a worldwide context. Forty-three linear variables were analyzed on crania derived from American, Asian, Australo-Melanesian, European, South-Saharan African, and Polynesian regions. South America was represented by seven Amerindian samples. In order to understand morphologic diversity among Amerindians of South America, variation was estimated using regions and local populations as units of analysis. Variances and F(ST) values were calculated for each unit, respectively. Both analyses indicated that morphologic variation in Southern Amerindians is extremely high: an F(ST) of 0.01531 was obtained for Southern Amerindians, and values from 0.0371-0.1205 for other world regions. Some aspects linked to the time and mode of the peopling of the Americas and various microevolutionary processes undergone by Amerindians are discussed. Some of the alternatives proposed to explain this high variation include: a greater antiquity of the peopling than what is mostly accepted, a peopling by several highly differentiated waves, an important effect of genetic drift, and gene flow with Paleoamericans. A combination of some of these alternatives explains at least some of the variation. PMID- 16044465 TI - Electron paramagnetic resonance analyses of surface radical chemistries of gamma sterilized orthopedic materials: implications pointing to cytotoxicity via wear debris-induced inflammation. AB - In this research, electron paramagnetic resonance (EPR) spin-trapping was utilized to determine if surface radical chemistries occur for gamma (gamma) sterilized orthopedic materials-ultra-high molecular weight polyethylene (UHMWPE) and the novel, hybrid, diurethane dimethacrylate (DUDMA)-based RHAKOSS. The materials' ability to competitively chelate catalytic ferrous ions (Fe(2+)) or readily reduce ferric ions (Fe(3+)), and hydrogen peroxide (H(2)O(2)) directly, in facilitating the Fenton reaction (FR), is indicative of cytotoxicity. Validations with a radical scavenger aids to confirm a radical mechanism. In conjunction, materials were thermally annealed and characterized by attenuated total reflectance-Fourier-transform infrared (ATR-FTIR) spectroscopy in order to explore accelerated oxidative degradation induced by residual radicals evolving from gamma-sterilization. Particularly, there was a significant decrease in spin adduct peak areas obtained from the reduction of H(2)O(2) in the presence of RHAKOSS or UHMWPE, evaluated against their respective controls. Additionally, chelated Fe(2+) accelerated the rate of FR. This phenomenon suggests that the materials are not better chelators than the Fe-activating chelator, edta. Neither material had the propensity to readily reduce Fe(3+) to the relevant Fe(2+), as certified by a nonradical mechanism. Alternatively, the false spin-adduct signal acquired when chelated Fe(3+) is employed arises via the nucleophilic addition of water onto the DMPO spin trap. Residual radicals in UHMWPE did not recombine/terminate following thermal annealing in an inert atmosphere. The radicals in RHAKOSS, however, did recombine under mild heating in an oxidizing or inert atmosphere. Both materials displayed quenching of ( )OH; however, for UHMWPE, this mechanism was jointly accountable for its accelerated degradation, evidenced by ATR-FTIR. Quenching of ( )OH by the silica found in RHAKOSS manifested in a competing effect that counterbalanced the observed FR. Implanted RHAKOSS is not likely to promote cytotoxicity and should not degrade, but the damaging effect of gamma sterilization on UHMWPE is a serious dilemma confronting its long-term durability and biocompatibility. PMID- 16044466 TI - Nitric oxide in wound-healing. AB - Modulation of the complex process of wound-healing remains a surgical challenge. Little improvement beyond controlling infection, gentle tissue handling, and debridement of necrotic tissue has been had in the modern era. However, increasing appreciation of the process from a biomolecular perspective offers the potential for making significant strides in wound modulation. The bioactive molecule nitric oxide was found to have wide-ranging impact on cellular activities, including the cellular responses engendered by wound healing. Current research suggests that nitric oxide and several nitric oxide donors can exert biologic effects, although the particular net responses of cells contributing to wound repair are context-dependent. PMID- 16044468 TI - Comparison of child morbidity in two contrasting medieval cemeteries from Denmark. AB - This study compares associations between demographic profiles, long bone lengths, bone mineral content, and frequencies of stress indicators in the preadult populations of two medieval skeletal assemblages from Denmark. One is from a leprosarium, and thus probably represents a disadvantaged group (Naestved). The other comes from a normal, and in comparison rather privileged, medieval community (AEbelholt). Previous studies of the adult population indicated differences between the two skeletal collections with regard to mortality, dental size, and metabolic and specific infectious disease. The two samples were analyzed against the view known as the "osteological paradox" (Wood et al. [1992] Curr. Anthropol. 33:343-370), according to which skeletons displaying pathological modification are likely to represent the healthier individuals of a population, whereas those without lesions would have died without acquiring modifications as a result of a depressed immune response. Results reveal that older age groups among the preadults from Naestved are shorter and have less bone mineral content than their peers from AEbelholt. On average, the Naestved children have a higher prevalence of stress indicators, and in some cases display skeletal signs of leprosy. This is likely a result of the combination of compromised health and social disadvantage, thus supporting a more traditional interpretation. The study provides insights into the health of children from two different biocultural settings of medieval Danish society and illustrates the importance of comparing samples of single age groups. PMID- 16044469 TI - High density cultures of embryonic stem cells. AB - Embryonic stem cells (ESC) have the unique ability to differentiate into a variety of tissue types. However, the realization of regenerative medicine will require the production of large quantities of ESC which subsequently have to be differentiated into the final phenotype. Thus, we sought to develop a simple and scaleable bioprocess to increase densities of ESC to achieve this goal. Using mouse embryonic stem cells (mESC) as a model, by combining automated feeding and culture of mESC on petriperm dishes, cell densities were enhanced up to 6.4 x 10(6) cells/cm2 compared to conventional petri dish culture which only reached 0.2 to 1.4 x 10(6) cells/cm2. It was found that mESC from all experiments maintained excellent viability, pluripotency, and genetic stability after growing for 6 days in petriperm cultures with automated feeding. The expression of Oct-4 transcription factor was observed in all cultures, mESC formed embryoid bodies in differentiated cultures and teratomas in SCID mice, confirming their pluripotency, and karyotype of the cultures was normal. This culture method was stable for routine passaging and a second mESC cell line was shown to perform in a similar manner on petriperm with automated feeding. This work represents an important step towards achieving high density cultures of ESC. PMID- 16044470 TI - Light-responsive bioconjugates as novel tools for specific capture of biologicals by photoaffinity precipitation. AB - Light-responsive bioconjugates are synthesized by a two-step protocol calling first for cotelomerization (chain-transfer polymerization) of N isopropylacrylamide and N-acryloxysuccinimide. The desired bioligand (biotin) is used in modified form as chain-transfer agent in this step. As a consequence, 100% of the produced bioconjugates carry this group. In a second step, the cotelomers (bioconjugates) are rendered photoresponsive by linking a chromophore ((3-aminopropyloxy)azobenzene) group to the N-acryloxysuccinimide side chains. The resulting structures show a critical solution temperature in pure water of 16 degrees C when the azo groups in the side chains are predominately in the (stable) trans state. Irradiation with UV light (330 nm) switches the azo group into the more hydrophilic cis state, and the critical solution temperature rises to 18 degrees C. Irradiation with visible light (> 440 nm) switches the group back to the trans state. Adjusting the temperature to an intermediate level, the bioconjugates are used to demonstrate the concept of photo affinity precipitation, i.e., the specific capture and recovery by light-induced precipitation of a target molecule (avidin) from a serum-containing cell-culture supernatant. The avidin was obtained in highly purified form; no nonspecific copurification of protein impurities was observable. PMID- 16044471 TI - Modeling of sodium acetate recovery from aqueous solutions by electrodialysis. AB - The main engineering parameters (i.e., ion transport numbers in solution and electro-membranes; effective solute and water transport numbers; effective membrane surface area, membrane surface resistances, and limiting current intensity) affecting the recovery of sodium acetate from model solutions by electrodialysis (ED) were determined in accordance with a sequential experimental procedure. Such parameters allowed a satisfactory simulation of a few validation tests carried out under constant or step-wisely variable current intensity. The performance of this ED process was characterized in terms of a current efficiency (omega) of about 93% in the constant-current region, a water transport number (t(W)) of about 15, and a specific energy consumption (epsilon) increasing from 0.14 to 0.31 kWh/kg for a solute recovery yield of 95% as the current density (j) was increased from 112 to 337 A/m2. The specific resistance of the anion- or cation-exchange membranes were found to be three or two times greater than those measured in aqueous NaCl solutions and are to be used to design and/or optimize ED stacks involved in the downstream processing of acetic acid fermentation broths. PMID- 16044472 TI - Abstracts of the 28th Annual Meeting of the American Society of Primatologists, Portland, Oregon, USA, August 17-20, 2005. PMID- 16044473 TI - Effects of methyl-beta-cyclodextrin-mediated cholesterol depletion in porcine sperm compared to somatic cells. AB - In this study, the use of methyl-beta-cyclodextrin (MBCD) to support capacitation of sperm cells was studied. Sperm were incubated with MBCD or alternatively capacitated in an in vitro fertilization medium. The effects of these incubations on phospholipid scrambling (using merocyanin), cholesterol depletion, GM-1 localization (using cholera-toxin B (CTX)), and membrane deterioration were assessed. For comparison, this was also tested in MBCD-treated MDCK cells. In MDCK cells, upto 71% of cholesterol was depleted, which coincided with a more diffuse CTX staining without any obvious effects on cell viability. In sperm, a similar depletion of 53% cholesterol was found after a 10 mM MBCD treatment. However, no merocyanin response was observed in viable sperm after MBCD treatments (indicating a lack of membrane changes associated with sperm capacitation). In contrast to MDCK, cells >1 mM MBCD caused plasma membrane disintegration and rendered sperm immotile. At higher concentrations also acrosome disruption was noted. CTX staining was absent at < 0.1 mM MBCD incubations but appeared at higher MBCD levels and was found to be specific for deteriorated cells that showed morphological signs of acrosome disruption. No significant plasma membrane deterioration, acrosome disruption, and sperm immotility nor CTX staining and only a modest (< 15%) cholesterol depletion were observed in conventionally capacitated sperm, where 40% of the intact sperm showed merocyanin staining. Taken together, the results indicate that membranes of sperm are more sensitive to MBCD-mediated cholesterol depletion than MDCK cells and that the use of MBCD to support sperm capacitation cannot be recommended due to its spermicidal effects. PMID- 16044474 TI - A [3]rotaxane with three stable states that responds to multiple-inputs and displays dual fluorescence addresses. AB - A [3]rotaxane molecular shuttle containing two alpha-cyclodextrin (alpha-CD) macrocycles, an azobenzene unit, a stilbene unit, and two different fluorescent naphthalimide units has been investigated. The azobenzene unit and the stilbene unit can be E/Z-photoisomerized separately by light excited at different wavelengths. Irradiation at 380 nm resulted in the photoisomerization of the azobenzene unit, leading to the formation of one stable state of the [3]rotaxane (Z1-NNAS-2CD); irradiation at 313 nm resulted in the photoisomerization of the stilbene unit, leading to the formation of another stable state of the [3]rotaxane (Z2-NNAS-2CD). The reversible conversion of the Z1 and Z2 isomers back to the E isomer by irradiation at 450 nm and 280 nm, respectively, is accompanied by recovery of the absorption and fluorescence spectra of the [3]rotaxane. The E isomer and the two Z isomers have been characterized by 1H NMR spectroscopy and by two-dimensional NMR spectroscopy. The light stimuli can induce shuttling motions of the two alpha-CD macrocycles on the molecular thread; concomitantly, the absorption and fluorescence spectra of the [3]rotaxane change in a regular way. When the alpha-CD macrocycle stays close to the fluorescent moiety, the fluorescence of the moiety become stronger due to the rigidity of the alpha-CD ring. As the photoisomerization processes are fully reversible, the photo-induced shuttling motions of the alpha-CD rings can be repeated, accompanied by dual reversible fluorescence signal outputs. The potential application of such light-induced mechanical motions at the molecular level could provide some insight into the workings of a molecular machine with entirely optical signals, and could provide a cheap, convenient interface for communication between micro- and macroworlds. PMID- 16044475 TI - Direct Mannich and nitro-Mannich reactions with non-activated imines: AgOTf catalyzed addition of pronucleophiles to ortho-alkynylaryl aldimines leading to 1,2-dihydroisoquinolines. PMID- 16044476 TI - Label-free electronic detection of thrombin in blood serum by using an aptamer based sensor. PMID- 16044477 TI - Solid-state structure and enantioselective reactions of a complex of a 1-thio substituted propargyllithium and (-)-sparteine. PMID- 16044478 TI - A zwitterionic spirocyclic pentacoordinate silicon compound synthesized in water by Si--O and Si--C bond cleavage. PMID- 16044479 TI - The "noncoordinating" anion Tf2N- coordinates to Yb2+: a structurally characterized Tf2N- complex from the ionic liquid [mppyr][Tf2N]. PMID- 16044480 TI - Formation of an unsymmetrical dinuclear ruthenium complex with mu-H, mu-OH, and mu-kappa2-CO2 bridges and multiple reactive sites. PMID- 16044481 TI - A revolution in electron microscopy. PMID- 16044482 TI - Artificial molecular double-stranded helices. PMID- 16044483 TI - Extension of the applicable range of fluorescein: a fluorescein-based probe for Western blot analysis. PMID- 16044484 TI - Redox catalysts for reduction with base metals. PMID- 16044485 TI - An ex vivo human model system to evaluate specificity of replicating and non replicating gene therapy agents. AB - BACKGROUND: Inefficiency, aspecificity and toxicity of gene transfer vectors hamper gene therapy from showing its full potential. On this basis significant research currently focuses on developing vectors with improved infection and/or expression profiles. Screening assays with validity to the clinical context to determine improved characteristics of such agents are not readily available since this requires a close relationship to the human situation. We present a clinically relevant tissue slice technology to preclinically test improved vector characteristics. METHODS: Slices were prepared from rat, mouse and human liver samples and from tumor tissue. Specificity of gene expression and replication was determined by infecting target and non-target tissue slices with transcriptionally retargeted adenoviruses and oncolytic viruses. RESULTS: Using rat liver slices, we demonstrate efficient knob-mediated adenoviral infectivity. A favorable tumor-on/liver-off profile, resembling in vitro and mouse in vivo data, was shown for a tumor-specific transcriptionally retargeted adenovirus by infecting slices prepared from tumor or liver tissue. Similar liver-off data were found for mouse, rat and human samples (over 3-log lower activity of the tumor specific promoter compared to cytomegalovirus (CMV)). More importantly, we show that this technology when applied to human livers is a powerful tool to determine aspecific replication of oncolytic viruses in liver tissue. A 2- to 6-log reduction in viral replication was observed for a tumor-specific oncolytic virus compared to the wild-type adenovirus. CONCLUSIONS: The precision-cut tissue slice technology is a powerful method to test specificity and efficiency of gene transfer as well as of viral replication using human tissue. PMID- 16044486 TI - [Neuropsychology of action and body schema]. PMID- 16044487 TI - [Survival after initial diagnosis of Alzheimer disease]. PMID- 16044488 TI - [Drop attacks in older adults: systematic assessment has a high diagnostic yield]. PMID- 16044489 TI - German biotech sector inches forward despite long odds. PMID- 16044490 TI - How to conduct competitive intelligence in your biotech startup. PMID- 16044491 TI - [Transposition osteotomy on the knee joint]. PMID- 16044492 TI - Do radiation doses below 1 cGy increase cancer risks? PMID- 16044493 TI - Radiation and screening. PMID- 16044494 TI - Joint effects of radiation and smoking on lung cancer risk among Atomic bomb survivors. PMID- 16044495 TI - Lung cancer after Hodgkin lymphoma: the roles of chemotherapy, radiotherapy and tobacco use. PMID- 16044496 TI - Residential radon, smoking and lung cancer. PMID- 16044497 TI - Lung cancer and plutonium exposure in Rocky Flat waters. PMID- 16044498 TI - Multiple gene effects in radiation oncogenesis. PMID- 16044499 TI - Multi-center screening of mutations in the ATM gene among women with breast cancer - the WECARE Study. PMID- 16044500 TI - The roles of BRCA1 and BRCA2 in the cellular response to ionizing radiation. PMID- 16044501 TI - The genetic basis for variation in radiation sensitivity in the general population. PMID- 16044502 TI - Gene environment interactions in a cohort of irradiated retinoblastoma patients. PMID- 16044503 TI - Second cancers after radiotherapy: any evidence for radiation-induced genomic instability? PMID- 16044504 TI - How important is ATM? PMID- 16044505 TI - Radioprotectors: current status and new directions. PMID- 16044506 TI - Radiation interactions with taxanes, old and new. PMID- 16044507 TI - The proteasome and radiation. PMID- 16044508 TI - Radiation-induced gene therapy. PMID- 16044509 TI - An American pioneer child neurologist and founder of neurological institutes. PMID- 16044510 TI - Recent publications in hematological oncology. PMID- 16044511 TI - Galanin receptors in the rat gastrointestinal tract. AB - Galanin functions are mediated by three distinct G-protein-coupled receptors, galanin receptor 1 (GalR1), GalR2 and GalR3, which activate different intracellular signaling pathways. Here, we quantified mRNA levels of GalR1, GalR2 and GalR3 in the gastrointestinal tract using real time RT-PCR. GalR1 and GalR2 mRNAs were detected in all segments with the highest levels in the large intestine and stomach, respectively. GalR3 mRNA levels were quite low and mostly confined to the colon. We also investigated the effect of galanin 1-16, which has high affinity for GalR1 and GalR2 and low affinity for GalR3 on depolarization evoked Ca2+ increases in rat cultured myenteric neurons using Ca2+-imaging. Intracellular Ca2+ changes in myenteric neurons were monitored using the Ca2+ sensitive dye, fluo-4, and confocal microscopy. Galanin 1-16 (1 microM) markedly inhibited the K+-evoked Ca2+ increases in myenteric neurons. In summary, the differential distribution of GalRs supports the hypothesis that the complex effects of galanin in the gastrointestinal tract result from the activation of multiple receptor subtypes. Furthermore, this study confirms the presence of functional GalRs and suggests that galanin modulates transmitter release from myenteric neurons through inhibition of voltage-dependent calcium channels involving a G(i/o)-coupled GalR. PMID- 16044512 TI - Translational repression by the cis-acting element of structure-anchored repression (CAESAR) of human ctgf/ccn2 mRNA. AB - The cis-acting element of structure-anchored repression (CAESAR) is a post transcriptional regulatory element of gene expression, which is located in the 3' untranslated region (UTR) of the human ccn2 gene (ctgf/ccn2). In this report, the repression mechanism of CAESAR, as well as the structural requirement, was investigated. Removal of minor stem-loops from CAESAR resulted in proportional attenuation of the repressive function, whereas removal of the single bulge or modification of primary nucleotide sequence did not affect its functionality. In light of functional mechanism, CAESAR exerted no significant effects on stability or nuclear export of the cis-linked mRNA. However, this element significantly interfered with the association of such mRNA on ribosome and slowed down the translation process thereafter in vitro. A translation repression mechanism by RNA secondary structure to determine the basal ctgf/ccn2 expression level was uncovered herein. PMID- 16044513 TI - Improving generalisation skills in a neural network on the basis of neurophysiological data. AB - The distribution of striate cortex cells exhibits a maximum number of cells tuned to vertical and horizontal orientations (Mansfield, 1974). This was interpreted as an adaptation of the visual system to the presence in the visual environment of greater amounts of vertical and horizontal information compared to information from other orientations (Keil & Cristobal, 2000). The present research confirms that vertical and horizontal orientations are, indeed, present in greater number in natural scenes. After normalization of the amount of information across all orientations, vertical information appeared to be better for bottom-up categorization. We demonstrate this using a connectionist autoassociator model of categorization used elsewhere in simulations of early infant categorization. PMID- 16044514 TI - Are carbonated beverages bad for you? PMID- 16044515 TI - Naps have gotten a bum rap. PMID- 16044516 TI - By the way, doctor. I'm 73, in generally good health, but bothered by frequent colds and respiratory infections. I suspect I have a weakened immune system. Are there tests that will tell me if that is so? PMID- 16044517 TI - Critics: U.S. government fails to prevent spread of HIV as rates rise. PMID- 16044519 TI - Rhode Island Senate passes medical marijuana bill. PMID- 16044518 TI - Cost imposes barrier to HIV treatment in Missouri. PMID- 16044520 TI - Involuntary HIV testing order upheld for sex offender. PMID- 16044521 TI - Inaugural conference on HIV, meth, hepatitis set for August. PMID- 16044522 TI - Global. HIV research company breaks language barriers. PMID- 16044523 TI - Mandated HIV test reversed for lack of body fluid exchange. PMID- 16044525 TI - Iowa's HIV criminal transmission statute survives challenge. PMID- 16044524 TI - Detainee denied HIV medicine loses claim against jailers. PMID- 16044526 TI - More PI drug interactions. PMID- 16044527 TI - A novel [99mTc[triple bond]N]2+ complex of metronidazole xanthate as a potential agent for targeting hypoxia. AB - A xanthate derivative (L) at the pendant hydroxy group of metronidazole, a nitroimidazole known to possess affinity for hypoxic tumors, has been used as the carrier molecule for targeted delivery of the gamma-emitting radioisotope 99mTc to tumors. The xanthate residues (S2(-)) from two molecules of this ligand (L) were used for chelation with the [99mTcN]2+ intermediate to form a square pyramidal and neutral [99mTcN/L2] complex in >95% yield using a low ligand concentration of 1 mg/mL (approximately 3 x 10(-3) M). Biodistribution studies carried out in Swiss mice bearing fibrosarcoma tumor showed selective accumulation of the injected activity in the tumor (1.44 +/- 0.26% per gram 1 h pi) with major clearance through hepatobiliary route. The complex showed high tumor/muscle ratio (2.15 and 3.35 at 1 and 3 h post-injection, respectively) and tumor/blood ratio, which were comparable to hypoxia targeting agents 99mTc BMS181321 and 99mTc-BRU59-21 reported earlier. PMID- 16044528 TI - Cognitive dysfunction following cardiopulmonary bypass and cerebral protection. PMID- 16044529 TI - Issues in Indian science. PMID- 16044530 TI - MR imagings of ciliated hepatic foregut cyst: an unusual cause of fluid-fluid level within a focal hepatic lesion (2005.4b). PMID- 16044531 TI - Germany plans to lure doctors back from west to east. PMID- 16044532 TI - Cambodia tackles high maternal mortality. PMID- 16044533 TI - Children choosing health. PMID- 16044534 TI - Warnings for asthma drugs. PMID- 16044535 TI - Lithium and valproic acid treatments reduce PKC activation and receptor-G protein coupling in platelets of bipolar manic patients. AB - Dysregulated protein kinase C (PKC) distribution and activation, and abnormal receptor-G protein coupling, have been implicated in the pathophysiology of bipolar affective disorder (BD). The therapeutic effectiveness of lithium has also been correlated with its ability to reduce PKC activation and G protein mediated signaling. We examine the cellular distribution and activation of PKC and receptor-G protein coupling in blood platelets from normal controls, patients with BD mania or schizophrenia during treatment-free state, and after lithium or valproic acid administration. PKC activity was measured under basal and 50 nM phorbol 12-myristate, 13-acetate (PMA), 1 microM serotonin or 0.5 U/ml thrombin stimulated conditions. The coupling of G proteins to serotonin or thrombin receptors were assessed by serotonin or thrombin-mediated [35S]GTPgammaS binding to membrane Galpha proteins. The results demonstrate that membrane-associated PKC activity and stimulus-induced PKC translocation are increased in BD manic, whereas stimulus-elicited PKC translocation is attenuated in schizophrenic patients. Lithium and valproic acid treatments attenuated the stimulus-induced PKC translocations to a similar degree and decreased PKC activity in both cytosolic and membranous fractions after two weeks of drug administration. An increase in 5-HT or thrombin stimulated [35S]GTPgammaS binding to Galpha proteins was detected in BD manic but not in schizophrenic patients although basal [35S]GTPgammaS binding was not different across the diagnostic groups. Lithium and valproic acid treatments similarly reduced receptor-G protein coupling with comparable time courses. Thus, increased membrane-associated PKC, cytosol to membrane PKC translocation and receptor-G protein coupling in platelets of BD manic patients were alleviated by lithium or valproic acid treatments. PMID- 16044536 TI - Formulation and evaluation of Alendronate Sodium gel for the treatment of bone resorptive lesions in Periodontitis. AB - Alendronate sodium is formulated into gels and evaluated for the treatment of bone resorptive lesions in periodontitis. Carbopol 934P was used for the preparation of gels in three different concentrations. The prepared gel was evaluated for various properties such as preformulation, content uniformity, viscosity, compatibility, sterility, in vitro diffusion, and in vivo studies. The drug and the polymer were found to be compatible and confirmed by Fourier transform infrared spectroscopy. Viscosity of the gels increased with the increase in the polymer concentration. The formulations were found to be sterile. In vitro release study revealed that drug released from the gel follows non Fickian diffusion followed by first-order release. In vivo studies were carried out for 6 months in patients. The results revealed a significant improvement in the clinical parameters such as gingival index, probing pocket depth, clinical attachment level, and potent inhibitory effect on bone resorption by inhibition of osteoclasts. In addition, there was increase in the new bone formation. PMID- 16044537 TI - Young brains on lead: adult neurological consequences? PMID- 16044539 TI - Modification of peptides and other drugs using lipoamino acids and sugars. AB - The scientific literature is full of new small molecules and larger peptides identified as potential pharmaceutical agents for a variety of diseases. The majority of these compounds, however, will never progress into the clinic because of poor oral absorption and low metabolic stability. The development of practical, economic, and widely applicable systems to improve the bioavailability of drugs is a highly sought-after goal. The conjugation of a drug with lipid and/or sugar units represents one of the most important strategies being investigated in this new field of drug delivery. This chapter describes one method of introducing lipidic groups to drugs via lipoamino acids and also provides useful procedures for the efficient incorporation of sugar units into drugs, particularly peptide drugs, via solid phase synthesis. PMID- 16044538 TI - Chimerism: a strategy to expand the utility and applications of peptides. AB - The modular nature of peptides can be exploited in the synthesis of chimeric sequences that combine diverse motifs in a single molecule. A theoretical consideration of the classification of peptides further expounds the multigeneric nature of peptide chimeras. Strategies for chimeric peptide syntheses include the chemical cross-linking of monomers and tandem combination by conventional SPPS. Additional details of chimeric peptide synthesis are also provided elsewhere in this volume. This chapter also explores some of the more common applications of chimeric peptides with particular emphasis on the molecular pharmacology of sequences that include address motifs for G protein-coupled receptors. Specific details of the biological properties of chimeras containing mastoparan, an amphiphilic tetradecapeptide component of wasp venom, further illustrate the novel and often unpredictable biological actions of chimeric constructs. These and numerous additional studies confirm that chimerism is an established strategy for the synthesis of molecular probes and bioactive agents. PMID- 16044540 TI - Synthesis of linear, branched, and cyclic peptide chimera. AB - Chimeric peptides are unnatural constructs consisting of bioactive compounds from at least two different peptide(s) and/or protein(s) or two sequences from different parts of the same protein. Such multifunctional peptide combinations are prepared to enhance the biological activity or selectivity of their components. New biological effects can also be achieved with the chimera. In this chapter the synthesis of three different types of chimeric peptides will be described. In a linear chimera, two peptide epitopes from different parts of glycoprotein D (gD) of herpes simplex virus (HSV) are combined. A branched chimera, built from linear peptides, consists of tuftsin oligomers with immunostimulatory activity and an epitope peptide of HSV gD. The third compound is a cyclic chimeric molecule, where alpha-conotoxin GI as a host peptide is modified by the incorporation of a core epitope from HSV gD as a guest sequence. PMID- 16044541 TI - Synthesis of cell-penetrating peptides for cargo delivery. AB - Cell-penetrating peptides (CPPs) have served as vehicles for the delivery of different molecules and particles into cells. The efficiency and noninvasive nature of peptide-mediated cellular transduction provides a promising tool for biomedical research. Studies of cell-penetrating constructs that deliver oligonucleotides, peptides, and proteins have elucidated several important cellular signaling mechanisms. Here we briefly describe the major classes of cell penetrating peptides. We also present various strategies used to couple different cargoes to CPPs. PMID- 16044542 TI - Incorporation of phosphotyrosyl mimetic 4-(phosphonodifluoromethyl)phenylalanine (F2Pmp) into signal transduction-directed peptides. AB - Phosphotyrosyl (pTyr)-containing sequences in proteins serve important roles in cellular signal transduction. Often, synthetic pTyr-containing peptides based on cognate sequences surrounding these pTyr residues can exhibit pharmacologically useful properties of full phosphoproteins. However, such pTyr-containing peptides have limited use in whole-cell systems resulting from lability of the phosphate ester bond to protein-tyrosine phosphatases (PTPs). For this reason, a number of phosphatase-stable pTyr mimetics have been developed that retain certain of the chemical and pharmacological properties of pTyr itself. Among these, difluoro phosphonomethyl phenylalanine (F2Pmp) has shown widespread utility in a variety of signal transduction settings, particularly those involving PTPs. This chapter provides practical techniques for the synthesis of a range of F2Pmp-containing peptides. PMID- 16044543 TI - Expressed protein ligation for protein semisynthesis and engineering. AB - Over the past decade, a significant methodological development in peptide ligation strategies has been elaborated that now permits the assembly of peptides and proteins. Native chemical ligation (NCL) has been introduced to join synthetic unprotected peptides by using the chemoselective reaction between a C terminal thioester and an N-terminal cysteine residue to result in a native peptide bond. Although this method has been applied to obtain peptides, small proteins, or protein domains (up to approx 150 residues), larger proteins could not been easily received because of the limited size of the ligated fragments. Intein technologies benefit from the opportunity to participate in the production of polypeptides with the reactive groups necessary for NCL aside from the rapid isolation of highly pure recombinant proteins. Expressed protein ligation extends the scope of NCL by overcoming the size limitation of target proteins accessible to synthesis. The intein splicing and EPL have been already proven to be useful for protein semisynthesis and for various investigations, including the studies of protein-protein interactions, segmental isotopic labeling for protein structure determination, synthesis of cytotoxic proteins, protein cyclization, and site-specific incorporation of noncanonical amino acids and biophysical probes into a protein sequence. PMID- 16044544 TI - Cellular delivery of peptide nucleic acid by cell-penetrating peptides. AB - Peptide nucleic acid (PNA) is a promising tool in biomedical research. PNA oligomers can be synthesized according to peptide synthesis protocols, but they hybridize to complementary RNA and DNA strands like oligonucleotides. The main hindrance to the use of PNA oligomers has been their poor uptake by cells. Fortunately, this limitation can be overcome by the application of peptide vectors. Different strategies are used to conjugate peptide vectors and PNA oligomers. In this chapter we discuss different types of delivery vectors (receptor ligands and cell-penetrating peptides) and strategies for the design and synthesis of PNA-vector conjugates. A protocol for the delivery of antisense PNA oligomer-cell-penetrating peptide conjugates is also described. PMID- 16044545 TI - Quenched fluorescent substrate-based peptidase assays. AB - The use of specific quenched fluorescent substrates (QFS) provides a rapid and sensitive method to measure peptidase activity, and is readily adaptable to high throughput screening of potential peptidase inhibitors. In this chapter, we discuss general considerations for the development of QFS assays, and describe in detail an assay protocol for the mammalian metallopeptidase, endothelin converting enzyme. PMID- 16044546 TI - A convenient method for synthesis of cyclic peptide libraries. AB - Cyclic peptides have been reported to bind to multiple, unrelated classes of receptor with high affinity. Owing to the robustness of amide bond chemistry, the ability to explore extensive chemical diversity by incorporation of unnatural and natural amino acids, and the ability to explore conformational diversity, through the incorporation of various constraints, arrays of cyclic peptides can be tailored to broadly sample chemical diversity. We describe the combination of a safety catch linker with a directed-sorted procedure for the synthesis of large arrays of diverse cyclic peptides for high-throughput screening. PMID- 16044547 TI - High-throughput peptide synthesis. AB - The methodologies of high-throughput peptide synthesis are overviewed and discussed. Particular focus is given to the techniques applicable to laboratories with a limited budget. Automated solutions for synthetic problems are also discussed. PMID- 16044548 TI - Backbone amide linker strategies for the solid-phase synthesis of C-terminal modified peptides. AB - This chapter describes backbone amide linker (BAL) strategies for the Nalpha-Fmoc solid-phase synthesis of C-terminal modified peptides. Most solid-phase protocols for the assembly of such peptides have limited generality, because they rely on the Calpha-carboxyl for attachment to the solid support. In the BAL approach, the growing peptide chain is anchored through a backbone nitrogen, thus allowing significant flexibility for chemical modification of the C-termini. In effect, any peptide containing C-terminal variations can be prepared in overall good purity and yield, with minimal side reactions, by using one or more of three variations (original and two modifications) of the BAL strategy. PMID- 16044549 TI - Synthesis of peptide bioconjugates. AB - Bioconjugates play an important role in several fields of biomolecular and biomedicinal sciences. Protein/polypeptide-based conjugates with covalently attached epitope peptides are considered as potential synthetic vaccine candidates and/or target antigens in affinity-based bioassays. This chapter describes the synthesis of two- and three-component bioconjugates using water soluble branched chain polymeric polypeptides with multiple amino and/or carboxyl groups as macromolecular partners and oligopeptides as epitopes with small molecular mass. The synthetic procedures outline three major strategies for the incorporation of multiple copies of uniformly oriented peptide epitopes. In the first example, chloroacetylated polypeptide is conjugated with SH-peptide to form a thioether linkage. Second, two independent oligopeptides are introduced into a macromolecule by amide and disulfide bonds, respectively. In the third example, a new procedure is reported for the formation of disulfide bridges by the use of Npys-modified polypeptide and SH-peptide. PMID- 16044550 TI - Protein identification by mass spectrometric analyses of peptides. AB - The focus of this chapter is the isolation and analysis of subsets of proteins or subproteomes. These methodologies can be applied to address fundamental questions in many areas of biological research. Protocols are provided for common techniques that include protein isolation from mammalian cells and the subsequent identification of peptides derived from endogenous proteins by the action of proteases. PMID- 16044551 TI - Manual solid-phase synthesis of glutathione analogs: a laboratory-based short course. AB - This chapter provides a manual for a laboratory-based short course to introduce the common techniques of solid-phase peptide synthesis (SPPS). The course provides students the opportunity to design and manually synthesize analogs of glutathione using relatively simple equipment available in any unsophisticated laboratory. The manual provides compact protocols for both the different steps of SPPS and the final cleavage of peptides from resin supports. We also introduce a simple method for the synthesis of combinatorial libraries of glutathione analogs that is suitable for those relatively unfamiliar with the field of peptide chemistry. PMID- 16044552 TI - Ecoregions and ecoregionalization: geographical and ecological perspectives. AB - Ecoregions, i.e., areas exhibiting relative homogeneity of ecosystems, are units of analysis that are increasingly important in environmental assessment and management. Ecoregions provide a holistic framework for flexible, comparative analysis of complex environmental problems. Ecoregions mapping has intellectual foundations in both geography and ecology. However, a hallmark of ecoregions mapping is that it is a truly interdisciplinary endeavor that demands the integration of knowledge from a multitude of sciences. Geographers emphasize the role of place, scale, and both natural and social elements when delineating and characterizing regions. Ecologists tend to focus on environmental processes with special attention given to energy flows and nutrient cycling. Integration of disparate knowledge from the many key sciences has been one of the great challenges of ecoregions mapping, and may lie at the heart of the lack of consensus on the "optimal" approach and methods to use in such work. Through a review of the principal existing US ecoregion maps, issues that should be addressed in order to advance the state of the art are identified. Research related to needs, methods, data sources, data delivery, and validation is needed. It is also important that the academic system foster education so that there is an infusion of new expertise in ecoregion mapping and use. PMID- 16044553 TI - Perspectives on the nature and definition of ecological regions. AB - Among environmental managers, recognition of the importance of integrating management activities across agencies and programs that have different responsibilities for the same geographic areas has created an awareness of the need for a common hierarchical framework of ecological regions (ecoregions) to implement the strategy. Responding to this need in the United States, nine federal agencies have signed a memorandum of understanding on the subject of developing a common framework of ecoregions. However, considerable disagreement over how to define ecoregions and confusion over the strengths and limitations of existing frameworks stand in the way of achieving this goal. This paper presents some perspectives on the nature and definition of ecoregions related to this confusion and provides a brief overview of the weight of evidence approach to mapping ecoregions, using an example initiated by the US Environmental Protection Agency. To effectively implement ecosystem assessment, management, and research at local, regional, and national levels, research is needed to increase our understanding of ecoregions. We must find ways to illustrate the nature of ecoregion boundaries and the variability of characteristics within ecoregions as they relate to management issues. Research must also be conducted on comparing existing frameworks and developing indices of ecological integrity to effectively evaluate their usefulness. PMID- 16044554 TI - Delineation and evaluation of hydrologic-landscape regions in the United States using geographic information system tools and multivariate statistical analyses. AB - Hydrologic-landscape regions in the United States were delineated by using geographic information system (GIS) tools combined with principal components and cluster analyses. The GIS and statistical analyses were applied to land-surface form, geologic texture (permeability of the soil and bedrock), and climate variables that describe the physical and climatic setting of 43,931 small (approximately 200 km2) watersheds in the United States. (The term "watersheds" is defined in this paper as the drainage areas of tributary streams, headwater streams, and stream segments lying between two confluences.) The analyses grouped the watersheds into 20 noncontiguous regions based on similarities in land surface form, geologic texture, and climate characteristics. The percentage of explained variance (R-squared value) in an analysis of variance was used to compare the hydrologic-landscape regions to 19 square geometric regions and the 21 U.S. Environmental Protection Agency level-II ecoregions. Hydrologic-landscape regions generally were better than ecoregions at delineating regions of distinct land-surface form and geologic texture. Hydrologic-landscape regions and ecoregions were equally effective at defining regions in terms of climate, land cover, and water-quality characteristics. For about half of the landscape, climate, and water-quality characteristics, the R-squared values of square geometric regions were as high as hydrologic-landscape regions or ecoregions. PMID- 16044555 TI - Using an ecoregion framework to analyze land-cover and land-use dynamics. AB - The United States has a highly varied landscape because of wide-ranging differences in combinations of climatic, geologic, edaphic, hydrologic, vegetative, and human management (land use) factors. Land uses are dynamic, with the types and rates of change dependent on a host of variables, including land accessibility, economic considerations, and the internal increase and movement of the human population. There is a convergence of evidence that ecoregions are very useful for organizing, interpreting, and reporting information about land-use dynamics. Ecoregion boundaries correspond well with patterns of land cover, urban settlement, agricultural variables, and resource-based industries. We implemented an ecoregion framework to document trends in contemporary land-cover and land-use dynamics over the conterminous United States from 1973 to 2000. Examples of results from six eastern ecoregions show that the relative abundance, grain of pattern, and human alteration of land-cover types organize well by ecoregion and that these characteristics of change, themselves, change through time. PMID- 16044556 TI - Toward a scientifically rigorous basis for developing mapped ecological regions. AB - Despite the wide use of ecological regions in conservation and resource management evaluations and assessments, a commonly accepted theoretical basis for ecological regionalization does not exist. This fact, along with the paucity of focus on ecological regionalization by professional associations, journals, and faculties, has inhibited the advancement of a broadly acceptable scientific basis for the development, use, and verification of ecological regions. The central contention of this article is that ecological regions should improve our understanding of geographic and ecological phenomena associated with biotic and abiotic processes occurring in individual regions and also of processes characteristic of interactions and dependencies among multiple regions. Research associated with any ecoregional framework should facilitate development of hypotheses about ecological phenomena and dominant landscape elements associated with these phenomena, how these phenomena are structured in space, and how they function in a hierarchy. Success in addressing the research recommendations outlined in this article cannot occur within an ad hoc, largely uncoordinated research environment. Successful implementation of this plan will require activities--coordination, funding, and education--that are both scientific and administrative in nature. Perhaps the most important element of an infrastructure to support the scientific work of ecoregionalization would be a national or international authority similar to the Water and Science Technology Board of the National Academy of Sciences. PMID- 16044560 TI - Implementing and auditing electronic recordkeeping systems used in scientific research and development. AB - Electronic recordkeeping is increasingly replacing handwritten records in the course of "normal business." As this trend continues, it is important that organizations develop and implement electronic recordkeeping policies and procedures. This is especially true for research and development organizations because of the potential to transform a discovery into a patent, and at times patent application contests are resolved in litigation. This paper provides a basis for the development, implementation, and subsequent assessment of a research and development recordkeeping policy. The approach described in this paper should be tailored by the organization adopting this approach to meet the needs of their organization. PMID- 16044561 TI - The historical impact of ISO 9000 on Lebanese firms. AB - The objective of this paper is to assess the impact of the new ISO 9001:2000 standard on the Quality Management Systems (QMS) of Lebanese firms that were already certified under ISO 9000:1994. To get an accurate feedback of stakeholders in a firm, three different questionnaires were developed and distributed to management, employees, and customers respectively. Empirical results indicate that ISO 9001:2000 has improved the QMS performance of Lebanese firms over that of the 1994 edition. Nevertheless, results showed that the new standard still has some weaknesses when it comes to improving suppliers' relationships and empowering employees. PMID- 16044562 TI - Nuts and your health: cracking old myths. PMID- 16044563 TI - What to do if Viagra won't do. PMID- 16044564 TI - On call. I'll be turning 65 in a few months. I understand that men my age should have a new test to check for bulging blood vessels. Can you explain the test and tell me if I should have it? PMID- 16044565 TI - Aspirin study refocuses prevention message for women. Aspirin therapy does less than anticipated in preventing heart attacks in women--but more than we knew in warding off strokes. PMID- 16044566 TI - A doctor talks about aspirin. PMID- 16044568 TI - 10 research-proven tips for better memory. Healthful habits help protect memory, but the aging brain may need an extra tweak or two to stay sharp. PMID- 16044567 TI - One more reason to get enough sleep. PMID- 16044569 TI - Health benefits of taking probiotics. We take vitamins and minerals to safeguard our health. Should we also add a daily dose of bacteria? PMID- 16044570 TI - By the way, doctor. I'm told I snore at night. I was completely unaware of it. Is snoring unhealthy? PMID- 16044571 TI - HIV and the lung in the HAART era. PMID- 16044572 TI - AIDS pandemic racing faster than means to stop it. PMID- 16044573 TI - HIV hope ride on therapeutic vaccine. PMID- 16044574 TI - What national policy shall we watch? PMID- 16044575 TI - Diagnostic utility of the subjective peripheral neuropathy screen in HIV-infected persons with peripheral sensory polyneuropathy. AB - A retrospective analysis of the Johns Hopkins University HIV neurology database was performed to assess the sensitivity and specificity of the Subjective Peripheral Neuropathy Screen (SPNS) for detecting HIV-associated neuropathies. The SPNS, a 3-item scale that evaluates lower extremity neuropathic symptoms, was administered to 75 patients from the HIV neurology outpatient clinic. Patients graded the severity of each symptom on a scale of 1 to 10, and the sensitivity, specificity, and diagnostic efficiency of the SPNS were calculated for each symptom. The results showed that the SPNS had a sensitivity of 47%, a specificity of 83%, a positive predictive value of 70%, and a diagnostic efficacy of 67%. The SPNS appears to be a useful screening tool for HIV-associated sensory neuropathies; it has a high specificity and a good positive predictive value. PMID- 16044576 TI - Editorial comment: diagnostic challenges of distal sensory polyneuropathy in HIV infected persons. PMID- 16044577 TI - Fanconi syndrome associated with use of tenofovir in HIV-infected patients: a case report and review of the literature. AB - We report a case of Fanconi syndrome associated with the use of tenofovir disoproxil fumarate, a nucleotide reverse transcriptase inhibitor used in the treatment of HIV infection. A 56-year-old HIV-infected man was admitted to the hospital with a chief complaint of severe, progressive weakness. His HIV infection was well controlled by antiretroviral therapy; other medical problems included hepatitis C and chronic renal insufficiency. About 2 weeks before presentation, the patient had received an influenza vaccination, which was followed by a generalized viral syndrome of several days' duration. Next, weakness developed and culminated in an inability to walk; this prompted the patient's presentation at the hospital. Urine chemistry, electrolyte panel, and clinical presentation were consistent with Fanconi syndrome, a generalized proximal tubular dysfunction involving proteins, glucose, uric acid, and electrolytes. Along with our Case Report, we review 25 cases of Fanconi syndrome previously reported in the literature. PMID- 16044578 TI - Editorial comment: tenofovir nephrotoxicity--vigilance required. PMID- 16044579 TI - Relationship between lipoatrophy and quality of life. AB - An informal yet comprehensive literature review of abstracts published in Medline was undertaken to identify papers that reported on how facial aging, lipodystrophy, and facial lipoatrophy affect quality of life. Facial lipoatrophy can erode self-esteem, cause psychological distress, and lead to depression. Persons with HIV infection encounter both stigmatization and marginalization as a result of facial lipoatrophy. In addition to exploring novel antiretroviral therapies that do not result in lipodystrophy, clinicians should consider treatments that correct the appearance of lipoatrophy for patients who feel adversely affected by the condition. PMID- 16044580 TI - Editorial comment: correcting facial lipoatrophy has little to do with vanity. PMID- 16044581 TI - Mechanisms of ensemble failure of the male gonadal axis in aging. AB - The systemic availability of testosterone (Te) falls by 35-50% after the sixth decade of life in healthy men. Intercurrent illness, trauma, surgery, stress, weight loss, diverse medications and institutionalization reduce Te concentrations further. Impoverishment of anabolic drive probably contributes to physical frailty and diminished quality of life in older individuals. However, the fundamental mechanisms that mediate hypoandrogenemia in the aging male remain unknown. Recent clinical experiments offer new clues to the mechanistic bases of Te depletion in older men. In particular, the following attributes mark aging related Te withdrawal: (a) high-frequency and low-amplitude LH pulses; (b) disorderly LH-release patterns, consistent with feedback disruption; (c) normal or heightened LH secretion following single or repeated GnRH stimuli; (d) reduced Te secretory-burst mass with normal basal Te secretion; and (e) impaired Leydig cell Te production in response to secreted LH pulses (stimulated by flutamide, tamoxifen, GnRH or anastrozole) and infused (recombinant human) LH pulses. The foregoing interconnected findings allow us to frame the integrative postulate that androgen deprivation in the older male reflects multisite failure in the GnRH-LH-testosterone axis. The most proximate locus of impairment is not yet known. PMID- 16044582 TI - Androgens and cognitive function. AB - Decrements in cognitive functioning are a common complaint among men with late onset hypogonadism (LOH). This article reviews the neurobiologic connection between gonadal steroids and cognitive functions, developmental effects and activational effects throughout the lifespan. Studies of hormone replacement therapy in older men are reviewed as well as epidemiological studies of endogenous hormones and cognition. While many studies of Androgen Replacement Therapy (ART) in older men suggest a potential beneficial effect on cognition, most studies to date have been small and need further replication with larger sample sizes. PMID- 16044583 TI - Downregulation of the major histocompatibility complex class I molecules by human herpesvirus type 8 and impaired natural killer cell activity in primary effusion lymphoma development. AB - Primary effusion lymphomas (PELs) are invariably infected by human herpesvirus type 8 (HHV8) and often co-infected by Epstein-Barr virus (EBV). We found that expression of major histocompatibility complex class I (MHC-I) surface molecules was significantly decreased in PEL cells when compared with HHV8 negative lymphomas, irrespective of EBV infection. MHC-I downregulation rendered PEL cells sensitive to recognition and killing by natural killer (NK) cells. Intriguingly, analysis of MHC-I non-restricted cytotoxicity in two PEL patients indicated a reduced NK cell activity when compared with healthy individuals. These data suggest that PEL outgrowth may require an impaired NK cell function. PMID- 16044584 TI - Decoding Ca2+ signals in plants. AB - Different input signals create their own characteristic Ca2+ fingerprints. These fingerprints are distinguished by frequency, amplitude, duration, and number of Ca2+ oscillations. Ca(2+)-binding proteins and protein kinases decode these complex Ca2+ fingerprints through conformational coupling and covalent modifications of proteins. This decoding of signals can lead to a physiological response with or without changes in gene expression. In plants, Ca(2+)-dependent protein kinases and Ca2+/calmodulin-dependent protein kinases are involved in decoding Ca2+ signals into phosphorylation signals. This review summarizes the elements of conformational coupling and molecular mechanisms of regulation of the two groups of protein kinases by Ca2+ and Ca2+/calmodulin in plants. PMID- 16044585 TI - Who's afraid of biotechnology? PMID- 16044586 TI - Animal campaigners pinpoint 'trivial experiments'. PMID- 16044587 TI - Swiss back animal research by a whisker. PMID- 16044588 TI - Code on medical ethics divides Poland's doctors. PMID- 16044589 TI - Ministers move to limit genome patents. PMID- 16044590 TI - Call for 'treaty' on human gene patents. PMID- 16044591 TI - How AIDS made patients act up. PMID- 16044592 TI - Danish trade unions oppose ban on screenings. PMID- 16044593 TI - Abandoned fraud case too baffling for jury. PMID- 16044594 TI - Writs fly over animal experiments. PMID- 16044595 TI - Mechanoreceptors rather than sedimentable amyloplasts perceive the gravity signal in hypergravity-induced inhibition of root growth in azuki bean. AB - Elongation of primary roots of azuki bean (Vigna angularis Ohwi et Ohashi) was suppressed under hypergravity conditions produced by centrifugation, such that the growth rate decreased in proportion to the logarithm of the magnitude of the gravity. The removal of the root cap did not influence the hypergravity-induced inhibition of root growth, although it completely inhibited the gravitropic root curvature. Lanthanum and gadolinium, blockers of mechanoreceptors, nullified the growth-inhibitory effect of hypergravity. These results suggest that the gravity signal for the hypergravity-induced inhibition of root growth is perceived independently from that of gravitropism, which involves amyloplasts as statoliths. Horizontal and basipetal hypergravity suppressed root growth as did acropetal hypergravity, all of which were nullified by the presence of lanthanum or gadolinium. These findings suggest that mechanoreceptors on the plasma membrane perceive the gravity signal independently of the direction of the stimuli and roots may utilise it to regulate their growth rate. PMID- 16044596 TI - The unconstitutional treatment of Nancy Cruzan. PMID- 16044597 TI - Dries v. Gregor: a case study of informed consent doctrine in New York. PMID- 16044598 TI - Radiative habitable zones in martian polar environments. AB - The biologically damaging solar ultraviolet (UV) radiation (quantified by the DNA weighted dose) reaches the martian surface in extremely high levels. Searching for potentially habitable UV-protected environments on Mars, we considered the polar ice caps that consist of a seasonally varying CO2 ice cover and a permanent H2O ice layer. It was found that, though the CO2 ice is insufficient by itself to screen the UV radiation, at approximately 1 m depth within the perennial H2O ice the DNA-weighted dose is reduced to terrestrial levels. This depth depends strongly on the optical properties of the H2O ice layers (for instance snow-like layers). The Earth-like DNA-weighted dose and Photosynthetically Active Radiation (PAR) requirements were used to define the upper and lower limits of the northern and southern polar Radiative Habitable Zone (RHZ) for which a temporal and spatial mapping was performed. Based on these studies we conclude that photosynthetic life might be possible within the ice layers of the polar regions. The thickness varies along each martian polar spring and summer between approximately 1.5 and 2.4 m for H2O ice-like layers, and a few centimeters for snow-like covers. These martian Earth-like radiative habitable environments may be primary targets for future martian astrobiological missions. Special attention should be paid to planetary protection, since the polar RHZ may also be subject to terrestrial contamination by probes. PMID- 16044599 TI - Sightseeing [far] beyond the Great Wall. PMID- 16044600 TI - The pregnant silence: Rust v. Sullivan, abortion rights, and publicly funded speech. PMID- 16044601 TI - Mandatory AIDS testing and privacy: a psycholegal perspective. PMID- 16044602 TI - Patient autonomy and state intervention: reexamining the state's purported interest. PMID- 16044603 TI - "Wrongful living": resuscitation as tortious interference with a patient's right to give informed refusal. PMID- 16044604 TI - Refusing life-sustaining treatment: can we just say no? PMID- 16044605 TI - Inmate fees for health care services. Final rule. AB - The Bureau of Prisons (Bureau) finalizes rules describing procedures we will follow for charging inmates fees for certain kinds of health services, as required under the Federal Prisoner Health Care Copayment Act of 2000 (Pub. L. 106-294, October 12, 2000, 114 Stat 1038, codified at 18 U.S.C. 4048). PMID- 16044606 TI - I have a conscience, too: the plight of medical personnel confronting the right to die. PMID- 16044607 TI - The state's interest in the preservation of life: from Quinlan to Cruzan. PMID- 16044608 TI - When altruism fails: reactions of unrelated bone marrow donors when the recipient dies. AB - This article examines the responses of bone marrow donors to the death of the unrelated person to whom they donated. Data analyzed were 330 questionnaires and fifty in-depth interviews collected from donors in the National Marrow Donor Program at one year post-donation. Death of the recipient produced feelings of guilt and responsibility in the donors in only a few cases (2% of donors from questionnaire data and 2 of the 23 donors interviewed). Grief occurred often (22 of 23 donors interviewed) and was often surprisingly intense, given the fact that the recipient was a stranger. Intensity of grief varied depending on the perceived relationship with the recipient. Our data indicate that limiting contact and/or information about the recipient to the donor would be unlikely to result in more positive psychosocial outcomes. However, several strategies which might be useful in relieving donor guilt and/or grief are suggested. PMID- 16044609 TI - Caution, contention, and consolidation. PMID- 16044610 TI - The constitution in a brave new world: a century of technological change and constitutional law. PMID- 16044611 TI - Guidelines for Catholic hospitals treating victims of sexual assault. PMID- 16044612 TI - Informed consent in Canada: an empirical study. AB - The case of Reibl v. Hughes has significantly altered the law regarding informed consent in Canada. It might be expected, therefore, that its impact on the Canadian medical profession would be significant. However, in the first study to examine the practice of Canadian doctors in this respect, Professor Robertson concludes that the profession is largely unaware of either the decision or its importance. Further, the study examines doctors' current perceptions and opinions on the disclosure of risks to patients and the results, among others, raise serious questions regarding both the awareness and reception of important legal rulings. PMID- 16044613 TI - Conversations with Rep. Ken Calvert. Interview by Frank Sietzen Jr. PMID- 16044614 TI - 2004 guidelines for surgical treatment of primary hepatocellular carcinoma. PMID- 16044615 TI - [Budd-chiari syndrome and heterozygotic mutation of metylenetetrahidrofolate reductase]. PMID- 16044616 TI - [Hepatoid adenocarcinoma of the stomach]. PMID- 16044617 TI - [Small bowel intussusception caused by metastatic cutaneous melanoma]. PMID- 16044618 TI - [Campylobacter jejuni bacteriemia associated with acute pancreatitis]. PMID- 16044619 TI - Contact sensitization in metalworkers. PMID- 16044620 TI - Proceedings of the International Symposium on Neuronal Migration Disorders and Childhood Epilepsies: clinical manifestations, pathomechanisms, and etiopathogenesis. April 16-17, 2004. Tokyo, Japan. PMID- 16044621 TI - Improving health, eliminating disparities. Proceedings of the Fourth Annual Primary Care and Prevention Conference. October 25-27, 2004. Atlanta, Georgia, USA. PMID- 16044622 TI - Respecting tribal traditions in research and publications: voices of five Native American nurse scholars. AB - A dialogue with five Native American scholars provides insight into conducting research and publishing resulting manuscripts on Native American topics, specifically healing beliefs and practices. This information provides a means to develop sensitivity and create understanding about concerns held by Native Americans regarding sharing certain defined cultural information with those outside the culture. The article identifies salient tribal issues related to research, discusses perspectives important to tribal nations and Native individuals surrounding research, and supplies a base on which to formulate further discussions. PMID- 16044623 TI - The knocked-out permanent tooth: information for patients, parents and caregivers. PMID- 16044624 TI - Influence of a fat layer on the near infrared spectra of human muscle: quantitative analysis based on two-layered Monte Carlo simulations and phantom experiments. AB - The influence of fat thickness on the diffuse reflectance spectra of muscle in the near infrared (NIR) region is studied by Monte Carlo simulations of a two layer structure and with phantom experiments. A polynomial relationship was established between the fat thickness and the detected diffuse reflectance. The influence of a range of optical coefficients (absorption and reduced scattering) for fat and muscle over the known range of human physiological values was also investigated. Subject-to-subject variation in the fat optical coefficients and thickness can be ignored if the fat thickness is less than 5 mm. A method was proposed to correct the fat thickness influence. PMID- 16044625 TI - Space shuttle flight environment induces degeneration in the retina of rat neonates. PMID- 16044626 TI - Antigenic protein in microgravity-grown human mixed Mullerian ovarian tumor (LN1) cells preserved in RNA stabilizing agent. PMID- 16044627 TI - Countermeasure for space flight effects on immune system: nutritional nucleotides. AB - Microgravity and its environment have adverse effects on the immune system. Abnormal immune responses observed in microgravity may pose serious consequences, especially for the recent directions of NASA for long-term space missions to Moon, Mars and deep Space exploration. The study of space flight immunology is limited due to relative inaccessibility, difficulty of performing experiments in space, and inadequate provisions in this area in the United States and Russian space programs (Taylor 1993). Microgravity and stress experienced during space flights results in immune system aberration (Taylor 1993). In ground-based mouse models for some of the microgravity effects on the human body, hindlimb unloading (HU) has been reported to cause abnormal cell proliferation and cytokine production (Armstrong et al., 1993, Chapes et al. 1993). In this report, we document that a nutritional nucleotide supplementation as studied in ground-based microgravity analogs, has potential to serve as a countermeasure for the immune dysfunction observed in space travel. PMID- 16044628 TI - Cell survival and preservation of siRNA-mediated protein knock-down upon serum free cryopreservation (-80 degrees C). PMID- 16044629 TI - Testing of Saccharomyces cerevisiae morphological fixatives and fixed samples stored at ambient temperature. PMID- 16044630 TI - Possible involvement of flow detection in the activation of osteoblasts. PMID- 16044631 TI - Polyamines protect against radiation-induced oxidative stress. AB - Astronauts and cosmonauts are exposed to a wide variety of different hazards while in space that include radiation, which presents one of the most critical barriers to long-term missions. A major deleterious effect directly associated with ionizing radiation is the production of reactive oxygen species (ROS) such as peroxides and hydroxyl radicals. The free radicals generated by ultraviolet (UV) or ionizing radiation can attack cellular lipids, proteins and DNA. Endogenous free radical scavengers such as glutathione and the polyamines (e.g, spermidine and spermine) can inhibit the action of ROS. In particular, heme oxygenase-1 (HO-1), the enzyme involved in heme protein metabolism, can provide antioxidant protection through the production of the antioxidant bilirubin. Furthermore, polyamines have been shown to indirectly increase HO-1 content and antioxidant protection. The beta2-adrenoceptor agonist clenbuterol has been shown to stimulate polyamine synthesis and by extension, might provide a margin of antioxidant protection through increasing HO-1 content. However, it is unclear whether the polyamines are acting as a tertiary messengers for antioxidant protection in the be beta2-adrenoceptor signal transduction pathway. The purpose of this study was to study the role of the polyamine pathway in attenuating free radical-induced damage. PMID- 16044632 TI - Erythropoietin and IL-3 receptor cell surface expression is decreased under conditions that model some aspects of microgravity. PMID- 16044633 TI - Secretion as a key component of gravitropic growth: implications for annexin involvement in differential growth. PMID- 16044634 TI - Phototaxis and aerotaxis in a calcifying alga. PMID- 16044635 TI - Co-expression and hormonal regulation of genes in response to gravity and mechanical stimulation in the Arabidopsis root apex. PMID- 16044636 TI - Alteration of growth and gravitropic response of maize roots by lithium. PMID- 16044637 TI - Gravity and light: integrating transcriptional regulation in roots. PMID- 16044638 TI - [Paediatric trauma]. PMID- 16044639 TI - [Introduction to evidence-based medicine]. PMID- 16044640 TI - [Laparoscopy in chronic abdominal pain in children]. AB - The recurrent chronic abdominal pain (RAP) is one of the most usual pathologies in pediatrics. It may appear in the form of periodical or continuous sharp crisis. In order to be accepted as chronic, the presence of this pain must last at least 3 months. In many of the reference publications, only between 5 and 1% of the cases are considered to have an organic ethiology and the rest are labelled as functional processes of psychological origin. The reason for our communication is to show our experience on 304 RAP patients from 1995 to 2001. We also want to highlight the usefulness of laparoscopy in order to diagnose and treat 14.45% (44) of cases of this group of patients. MATERIAL AND METHODOLOGY: 304 patients between 5 and 14 years old were studied from 1995 to 2001. We followed a protocol of: medical and personal medical record, specific anamnesis for RAP, detailed description of the diet and intestinal habits, physical exploration. Laboratory: 1) blood analysis, hepatic profile, glucose, cholesterol, creatinine, amylase, 2) urine sediment and may be urine culture, 3) faeces analysis and 4) vaginal flow analysis. Image diagnosis is: simple abdomen radiography, ultrasonography and intestinal Rx. Guided and specific determinations: spired urea test, gastric chemism, lactose test, gastroscopy and biopsy, colonoscopy and biopsy, laparoscopy. RESULTS: In short, we can say: 74 patients (23.4%) come with inappropiate feeding diets, 31 (10.1%) with helicobacter pilori, 20 (6.5%) with adenoids, pharynx and pharyngeal processes, the same figure from gynaecologic origin, 18 (5.9%) from psychological origin, and in 44 cases (14.45%) laparoscopy was indicated. CONCLUSIONS: The anatomopathological study in the cases in which we carried out laparoscopic appendicectomy does not allow us to accept the symptoms of chronic appendicitis. The exploratory laparoscopy and the appendectomy have meant benefits for the solution to this pathology in 14.45% of patients. It is essential to carry out a long-term follow-up, at least 12 months, to be sure RAP is cured. We must carry on looking for aetiological cause of RAP. PMID- 16044641 TI - [Minimally invasive approach of Nuss for the correction of pectus excavatum]. AB - Since December 1999 to November 2003, we treated 68 patients with pectus excavatum using the Nuss minimally invasive procedure. Their ages were between 6 and 24 years old, with a mean of 12 years. Average Haller index was 4.1. Lateral stabilisers were used in all the cases, in 28% of them, two stabilisers were used. Mean operative time was 70 minutes (range 40 to 120). A thoracic epidural catheter was used for postoperative analgesia in 87% of the cases, with a mean duration of 3 days. The most frequent early complication was residual pneumothorax, which resolved spontaneously in 24% of the cases. The average hospital stay was 5.7 days. 93% of patients had excellent, very good or good results, in 5% of the cases the results were fair, and 2% had poor results due to thoracic asimetry and/or sternal rotation. The Nuss minimally invasive procedure is a useful method for treatment of pectus excavatum's patients. PMID- 16044642 TI - [Retroperitoneoscopic heminephrectomy]. AB - The practice of minimally invasive surgery techniques in the retro-peritoneal space presents additional difficulties. Due to the absence of a real cavity and the presence of a dense fat tissue involving the structures, the need for an artificial space to work in is necessary. We present three cases of the retro peritoneal superior heminephrectomy present in a 3-months, 6-months and a 6-year old children. In all cases a 1 centimeter port for the optique and two 0.5 centimeter ports for the instruments were situated, and the use of harmonic scalpel was satisfactory to remove renal tissue without bleeding. Drainages were unnecessary and all of the children left the hospital within the first 48 post operative hours. Since 1994, when Figenhans and Clayman communicated the first laparoscopic nephrectomy in children, the technique has been used generally and their forward advances in conventional surgery are now universally accepted. Furthermore, the practice of polar retro-peritoneoscopic heminephrectomy allowed a very precise renal hiliar dissection and an excellent differentiation of each hemirenal tissue, separating the ill kidney and preserving the maximum function possible for the other half-kidney and the adrenal gland. PMID- 16044643 TI - [Management of choledochal cyst: laparotomy or endoscopic]. AB - The most generally accepted therapy of choledochal cyst is cystectomy and biliar derivation by laparotomy. Last years, endoscopic papilotomy by ERCP has been a valuable therapeutic alternative, no only a diagnostic method. In this study, we reviewed five pediatric patients operated in our Deparment in last five years for choledochal cyst. The initial therapy was laparotomy (n=4) and endoscopic papilotomy by ERCP (n=1) This one was made in other Hospital. Follow-up has been between one and five years. All patients are living. Four patients who were operated by laparotomy are asyntomatic. Patient who was treated by ERCP needed a new ERCP in first posoperative month. Five years ago, she had a seriuos acute pancreatitis and we decided laparotomy and biliar derivation. Since laparotomy, she had two new episodes of acute pancreatitis and she has needed a new endoscopic dilatation with ballon by ERCP. She has been asyntomatic for four months. In conclusion, we think laparotomy with biliar derivation is safer than ERCP in management of children with choledochal cyst. ERCP must be reserved to emergency situations before laparotomy or after postoperative complications, never as exclusive therapy. PMID- 16044644 TI - [Parapneumonic effussion. A review of 33 cases over 6 years]. AB - The incidence of empyema thoracis in children seems to be increasing. The objective [corrected] of this study is to propose a rational initial treatment of parapneumonic effusions in order to reduce hospital stay and late-stage empyema complications. Medical records of 33 children presenting parapneumonic effusions who required any surgical therapy between 1997 and 2002 were reviewed. They were grouped as (I) successful management with chest tube, (II) successful management with intrapleural instillation of urokinase and (III) successful surgical treatment: (IIIa) thoracoscopy or (IIIb) thoracotomy. CONCLUSION: Early sonographic evaluation of parapneumonic effusions is usefull to evaluate the severity of disease and the need for surgical intervention. Thoracoscopic assisted surgery is an effective treatment for pediatric early-stage empyema. Thoracotomy is indicated for most children with established empyema. PMID- 16044645 TI - [Laparoscopic approach for treatement of gastroesophageal reflux in newborns and infants with severe respiratory disorders. When to indicate it?]. AB - Gastroesophageal reflux (GER) is a common finding in the newborn period, which is in general well tolerated, and tends to resolve spontaneously at about 18 months of age. However, a small number of children will present important respiratory manifestations (aspiration and apneic episodes), or increase in the manifestations of pre existing conditions (tracheomalacia, laryngomalacia), which can produce life threatning events. Ph monitoring does not completely discard the possibility of GER and the use of prokinetic drugs, in association with other drugs to protect the esophagus do not prevent serious consecuences produced by reflux to the airway. In this report six children with severe airway disease and marked GER, with no response to initial medical treatment required laparoscopic fundoplication. They have all resolved their airway disease, and are currently well. The laparocopic technique for fundoplication of small children seems to be a secure, well tolerated and definitive treatment in the hands of trained surgeons. PMID- 16044646 TI - [Role of peritoneal drainage in very low birth weight with enterocolitis]. AB - Peritoneal drainage (PD) has been proposed as a temporizing procedure for perforated necrotizing enterocolitis (NEC) in very low birth weight neonates. This operation was designed for patients considered too unstable to undergo laparotomy (LAP). Since the introduction of PD some investigators have suggested that it may serve as a definitive therapy. The aim of our study is to determine the efficacy of PD for the stabilization of patients with complicated NEC and its utility as a definitive surgical treatment. METHODS: We review the clinical records from all the patients treated in our unit because of NEC that required surgical therapy. We analyze the mean blood pressure, cardiac and respiratory rate, inspired oxygen concentration, mean airway pressure, diuresis, and arterial blood gases measured 6 and 12 hours after the initial surgical treatment. We divide patients in two different groups according to the initial surgical procedure, peritoneal drainage (PD) or laparotomy group (LAP). Data is shown as media +/- standard deviation, statistical analyses were performed using analysis of variance (ANOVA) for repeated measures and Mann-Withney test. RESULTS: From January 1997 to January 2001 we treated 13 patients with necrotizing enterocolitis that required surgical treatment. Among this group 6 patients were managed initially with PD and 7 with LAP. The gestational age media was 29.07 +/- 3.81 weeks and the birth weight mean 1199.76 +/- 521 gr., without any significant differences between the two groups DP and LAP. All patients improved haemodynamically and respiratory 6 and 12 hours after the surgical treatment. Nevertheless, the mean blood pressure improved even more in the DP group (p<0.005). The DP group showed an improvement in all parameters 6 hours after the drainage was placed, but this effect wasn't maintained for the next 12 hours onwards. From the 12 hours after PD all patients suffered a steady worsening that required further surgical procedures. The overall mortality was 3 patients (23%), without differences between the two groups. The surgical techniques performed (bowel resection, diversion) were similar for both groups. DISCUSSION: DP allows the stabilization for very critically ill patients with complicated NEC. However, this stabilization is temporary. This improvement lasts for a few hours providing a better status for the definitive surgical treatment for the perforated NEC. In our experience DP could not be considered as a definitive surgical treatment. PMID- 16044647 TI - [Evidence-based medicine in Spanish pediatric surgery reports]. AB - INTRODUCTION: Pediatric surgical literature is characterized by a high prevalence of observational studies and a paucity of randomized controlled trial, comparing mainly medical therapies. AIMS: To asses scientific evidence level produced by spanish pediatric surgeons. MATERIAL AND METHODS: Articles reported in Cirugia Pediatrica, surgical reports in Anales Espanoles de Pediatria, and reports from spanish authors published in the Journal of Pediatric Surgery, Pediatric Surgery International and European Journal of Pediatric Surgery, from 1997 to 2002. Internal validity was assessed through the critial guides from Sackett, and evidence level was classified through the Oxford Centre for Evidence Based Medicine classification. RESULTS: 94.5% on therapy reports, 61.5% on diagnosis, 83.3% on prognosis and 52.2% on risk factors or etiology, were classified as level 4 (descriptive studies). Only four randomized controlled trials were found, all of them comparing medical therapies (analgesia, antibiotics and surgical stress control). CONCLUSIONS: Our results are similar to those offered by previous reviews, showing up a low level of scientific evidence in most articles. Diffusion of guides on reporting descriptive studies may improve the evidence level of our reports. PMID- 16044648 TI - [Spigelian hernia: two cases associated to cryptorchidism]. AB - Spigelian hernia (SH) is rare in children, it occurs primarily en adults the mean age of presentacion is 50 years. We present two patients with hernia associated to cryptorchidism one of them bilateral. PMID- 16044649 TI - [Wandering spleen torsion]. AB - The ectopic spleen characterizes for absence of its suspensory ligaments and a long pedicle that are predisposed to complicate it for a torsion with commitment of the venous drainage at first and arterial at a later time himself, producing increase of its volume and infarct. This anomalous situation, it can be had to a congenital malformation of the development of its suspensory elements and fall toward the inferior abdomen or else to an inferior growth of the mesodermic yolk of that this organ originates itself. The suitable treatment is the laparoscopic splenopexy, but when it exists infarction, it is no possible avoid the splenectomy. PMID- 16044650 TI - The oracle of cough. PMID- 16044651 TI - Overactive bladder. PMID- 16044652 TI - A 35-year old man with recurrent hoarseness. PMID- 16044653 TI - Primary percutaneous coronary intervention for acute MI: improving access and outcomes. AB - Patients with acute myocardial infarction (MI) with ST-segment elevation have better outcomes with primary percutaneous coronary intervention (PCI) than with fibrinolytic therapy. Multiple clinical trials in the past 10 years have addressed ways to improve PCI as primary therapy for acute MI. Logistic strategies to improve access to PCI are being studied. PMID- 16044654 TI - Imaging in acute brain infarction. AB - Imaging in a patient suspected of having suffered an acute stroke is used primarily to clear the patient for thrombolytic therapy, by excluding intracranial hemorrhage and nonstroke causes of the patient's symptoms, within a critical 3-hour window. Noncontrast CT of the brain is the imaging test of choice for the initial evaluation of a patient with suspected acute ischemic stroke. It is rapid and readily available and has a high sensitivity for intracranial hemorrhage. Contrast-enhanced CT angiography and perfusion imaging may provide additional information with only a minimal increase in scanning time. Finally, diffusion-weighted magnetic resonance sequences have a high sensitivity for acute stroke and, with magnetic resonance perfusion imaging, may play an increasing role in the future treatment of stroke. PMID- 16044655 TI - Gait disorders: search for multiple causes. AB - Gait disorders predict functional decline in older adults. They are often the result of multiple causes, so a full assessment should consider different sensorimotor levels and should include a focused physical examination and evaluation of functional performance. Exercise and medical and surgical interventions are effective and can reduce the degree of gait disorder, but usually not without some residual impairment. Orthoses and mobility aids are also important interventions to consider. PMID- 16044656 TI - Pertussis: old foe, persistent problem. AB - Although a safe and effective vaccine is available, pertussis continues to be an important cause of morbidity and mortality. Immunity acquired from natural infection or vaccination wanes within 5 years, making older children, adolescents, and adults important reservoirs of infection. Many neonates and infants contract pertussis from older people with mild symptoms and are at risk for developing severe, life-threatening illness. Immunization programs are being considered for adolescents and for adults who live with or care for infants. PMID- 16044657 TI - Physician's guide to the new 2005 dietary guidelines: how best to counsel patients. AB - The Dietary Guidelines for Americans 2005 encourage most Americans to eat fewer calories, be more active, and make wiser food choices. Health care providers can influence patients' food and activity choices by providing specific counseling and presenting straightforward information. PMID- 16044658 TI - Nutrition: know the facts. PMID- 16044659 TI - Recognizing and managing acute diverticulitis for the internist. AB - Colonic diverticulitis can present as mild abdominal discomfort or as life threatening septic shock and can also mimic many inflammatory conditions. Most patients with acute diverticulitis can be managed with antibiotics and supportive care. Surgery is reserved for those with repeat attacks, disease unresponsive to medical therapy, certain complications, and the possibility of colon cancer that cannot otherwise be excluded. PMID- 16044660 TI - Editorial overview: serotonergic agents in functional GI disorders: targeting the brain-gut axis. PMID- 16044661 TI - The potential role for statins in the treatment of multiple sclerosis. AB - In 1995, it was observed that the administration of statins to heart transplant patients resulted in fewer episodes of rejection, thus a role for statins in the treatment of inflammatory disease was suggested. To date, the results of a single, open-label trial in multiple sclerosis patients have demonstrated that treatment with one of the statins, simvastatin, reduced the number and volume of lesions, as observed using gadolinium-enhancing magnetic resonance imaging. While the results of this first study seem promising, the rationale for using a cholesterol-lowering drug in a demyelinating disease must be addressed, in addition to the potential problems that the side effects of statins may produce in multiple sclerosis patients. PMID- 16044662 TI - The endocannabinoid system: a drug discovery perspective. AB - The endocannabinoids are lipid messengers that engage the same cell surface receptors targeted by delta9-tetrahydrocannabinol, the active component of marijuana. They are produced by cells in the brain and other tissues and combine with two subtypes of G protein-coupled cannabinoid receptors, CB1 and CB2. Their ability to modulate a variety of pathophysiological processes, including appetite, pain and mood, provides unique opportunities for drug discovery. Three such opportunities are discussed here: reduction of body weight through blockade of CB1 receptors, alleviation of pain through activation of extracerebral cannabinoid receptors, and modulation of pain and anxiety through inhibition of endocannabinoid degradation. PMID- 16044663 TI - Cannabinoids as potential anti-epileptic drugs. AB - Cannabinoids have long been recognized as having the potential for both anticonvulsant and proconvulsant effects. The increased understanding of the cannabinoid receptors and their endogenous ligands over the last decade has provided a potential mechanism of action for these apparently paradoxical effects. Although the anticonvulsant effects of cannabinoids appear to be mediated by their action at presynaptic cannabinoid receptors, which inhibit the release of excitatory neurotransmitters such as glutamate, it is clear that they are also capable of producing proconvulsant effects through the activation of cannabinoid receptors on terminals releasing inhibitory neurotransmitters, such as gamma-amino-butyric acid. In the brain, the activation of cannabinoid receptors is carefully controlled by the rapid synthesis and degradation of endocannabinoids in a way that targets the endogenous ligands to specific sets of cannabinoid receptors. The potential problem in delivering a cannabinoid drug to treat epilepsy is the inability to control its actions at different cannabinoid receptors regulating the release of different neurotransmitters. Since the action of cannabinoids is complex, and there is a dearth of clinical trial data, it is currently unclear whether cannabinoids might be both efficacious and safe in the treatment of epilepsy. PMID- 16044665 TI - Astrocytes as targets for neuroprotective drugs. AB - Astrocytes have long been considered as merely structural support for neurons in the central nervous system (CNS). However, more recent evidence has demonstrated roles for astrocytes in both neuroprotection and neurodegeneration. One major role is in the modulation of glutamatergic neurotransmission. Astrocytes can also modulate inflammatory responses in the CNS; they contain high levels of antioxidants and mediate neuronal protection after physiological assault, as well as releasing neurotrophic factors and being important to neuronal plasticity in the hippocampus. Targeting astrocytes as mediators for neuroprotective drugs may be a promising strategy. PMID- 16044664 TI - The ubiquitin-proteasome system as a drug target in cerebrovascular disease: therapeutic potential of proteasome inhibitors. AB - Proteasomes are large, multi-catalytic protease complexes that are found in the cytosol and in the nucleus of eukaryotic cells with a central role in cellular protein turnover. The ubiquitin-proteasome system (UPS) is the predominant non lysosomal protein degradation pathway that ensures the viability, proliferation and signaling of eukaryotic organisms. Overwhelming data exist implicating a critical role for the UPS in cerebral ischemic injury. Ischemic and hypoxic trauma, and their associated oxidative, nitrosylative and energetic stress, underlie neurodegeneration following stroke, and evoke a discreet set of transcriptional events which have a complex and interdependent relationship with proteasomal function. Rapid elimination of denatured, misfolded and damaged proteins by the proteasome becomes a critical determinant of cell fate. Proof-of principle has been obtained from animal models of cerebral ischemia, in which proteasome inhibitors reduce neuronal and astrocytic degeneration, cortical infarct volume, infarct neutrophil infiltration. and nuclear factor kappaB immunoreactivity. This neuroprotective efficacy has also been observed when proteasome inhibitors have been used 6 h after ischemic insult. Strategies aimed at effecting long-lasting changes in proteasomal function are not recommended, given the growing body of evidence implicating long-term proteasomal dysfunction in chronic neurodegenerative disease. These effects are likely due to the fact that the UPS is also essential for cellular growth, metabolism and repair, and untoward effects of proteasomal inhibition indicate that the development of short lived proteasome inhibitors, or compounds which can spatially and temporally regulate the UPS, is a desirable clinical target. Studies in animal models indicate that the use of specific proteasome inhibitors may be beneficial in treating a host of acute neurological disorders, including ischemic stroke. PMID- 16044666 TI - A potassium channel, the M-channel, as a therapeutic target. AB - Compounds that stimulate or inhibit M-channels (ie, voltage-gated potassium channels formed by KCNQ2, KCNQ3 and KCNQ5) have been evaluated in clinical trials for epilepsy, stroke and Alzheimer's disease. The importance of M-channel function in reducing neuronal excitability is underscored by the finding that KCNQ2/3 mutations causing mild reduction of M-channel activity are linked to neonatal epilepsy. M-channel openers decrease the hyperexcitability responsible for epileptic seizures, neuropathic pain and migraine. Conversely, M-channel blockers may enhance cognitive functions. The M-channel has thus emerged as a promising target for treating epilepsy, stroke, migraine, pain, dementia, anxiety and bipolar disorder. PMID- 16044667 TI - Drug treatments for subjective tinnitus: serendipitous discovery versus rational drug design. AB - Progress has been made in understanding the neural basis of subjective tinnitus (ST); however, this has not, as yet, translated into many new drug treatments. One reason for this is that realistic behavioral models of ST in animals have been developed only recently, and are still not widely used. Nonetheless, some significant pharmacological advances have been made. At present, there is evidence to support the efficacy of transtympanic gentamicin administration in the treatment of tinnitus associated with Meniere's disease; there is also some evidence to support the efficacy of intratympanic steroid and lidocaine application in the management of ST. Although benzodiazepines and anti-epileptic drugs appear to be effective in many cases of this condition, there is concern about their adverse side effect profile. Based on well-controlled clinical trials, vasodilators such as misoprostol, and histamine receptor ligands should be further investigated. Finally, given the evidence that ST is a form of sensory epilepsy, new antiepileptic drugs should be tested for potential efficacy as they are developed; such drugs may include novel N-methyl-D-aspartate receptor antagonists, as well as cannabinoids. PMID- 16044668 TI - Talnetant GlaxoSmithKline. AB - Talnetant (SB-223412) is a selective, orally active NK3 antagonist based on 4 quinolinecarboxamide, and is under development by GlaxoSmithKline (formerly SmithKline Beecham) for the potential treatment of several disorders, including urinary incontinence, irritable bowel syndrome and schizophrenia. By November 2004, the compound had completed phase II trials. PMID- 16044669 TI - Spheramine Titan/Schering. AB - Spheramine, which is composed of microcarriers coated with dopamine-producing human retinal pigment epithelial cells, is being developed by Titan and Schering for the potential treatment of advanced Parkinson's disease. Phase II trials were ongoing in March 2005 and, at this time, were expected to be completed in the second half of 2006. PMID- 16044670 TI - Phenserine Axonyx. AB - The acetylcholinesterase inhibitor phenserine, the (-)-enantiomer of a phenylcarbamate derivative of physostigmine, is being developed as a potential therapy for Alzheimer's disease by Axonyx, under license from the National Institutes of Health/National Institute on Aging. In March 2005, Axonyx suspended patient recruitment for the ongoing phase III trials of phenserine, after the drug failed to meet the primary endpoints of the first of these trials, but in May 2005, the company was planning to reformulate the drug. PMID- 16044671 TI - Brivaracetam UCB. AB - Brivaracetam (UCB-34714), an orally active, high-affinity synaptic vesicle protein 2A ligand, is being developed by UCB for the potential treatment of neurological disorders, namely epilepsy and neuropathic pain. By December 2003, UCB reported that phase II epilepsy trials had begun [517569]. PMID- 16044672 TI - NBI-5788 Neurocrine. AB - Neurocrine Biosciences is developing NBI-5788, an analog of an immunodominant epitope of myelin basic protein for the potential intravenous treatment of multiple sclerosis. By April 2005, enrollment for phase II trials was complete, with results expected early in 2006. PMID- 16044673 TI - Disease mapping and risk assessment in veterinary parasitology: some case studies. AB - Disease mapping and risk assessment are important tasks in the area of medical and veterinary epidemiology. The development of methods for mapping diseases has progressed considerably in recent years. Geographical Information Systems (GIS), Remote Sensing (RS), and Spatial Analysis represent new tools for the study of epidemiology, and their application to parasitology has become more and more advanced, in particular to study the spatial and temporal patterns of diseases. The present review highlights the usefulness of GIS and RS in veterinary parasitology in order to better know the epidemiology of parasite organisms, causing either snail/arthropod borne diseases or direct transmissible diseases, mostly in small areas with a strong impact by man. It demonstrates the potential of these technologies to serve as effective tools for: data capture, mapping and analysis for the development of descriptive parasitological maps; studying the environmental features that influence the distribution of parasites; predicting parasite occurrence/seasonality based on their environmental requirements and as decision support for disease intervention; and surveillance and monitoring of animal diseases. PMID- 16044674 TI - Biology-based mapping of vector-borne parasites by Geographic Information Systems and Remote Sensing. AB - Applications of growing degree day-water budget analysis and satellite climatology to vector-borne parasites are reviewed to demonstrate the value of using the unique thermal-hydrological preferences and limits of tolerance of individual parasite-vector systems to define the environmental niche of disease agents in the landscape by modern geospatial analysis methods. PMID- 16044675 TI - Advances in satellite remote sensing of pheno-climatic features for epidemiological applications. AB - Geographical Information Systems (GIS) and Remote Sensing (RS) technologies are being used increasingly to study the spatial and temporal patterns of diseases. They can be used to complement conventional ecological monitoring and modelling techniques, and provide a means to portray complex relationships in the ecology of diseases with strong environmental determinants. In particular, satellite technology has been extraordinarily improved during recent years, providing new parameters useful to understand the epidemiology of parasites, such as vegetation indices, land surface temperatures, soil moisture and rainfall indices. In the present review, Normalized Difference Vegetation Index (NDVI) is primarily considered, since it is the index characterizing vegetation that is most used in epidemiological studies. Multi-temporal study of RS data allows collection of bio climatic information about risk area distribution, along with predictive studies and anticipatory models of diseases, at different geographic scales ranging from global to local. The main physical and technological basis of a mathematical model, effective at different scales, for identification of landscape pheno climatic features is described in the current paper. PMID- 16044676 TI - Sizing up human health through remote sensing: uses and misuses. AB - Following the launch of new satellites, remote sensing (RS) has been increasingly implicated in human health research for thirty years, providing a growing availability of images with higher resolution and spectral ranges. However, the scope of applications, beyond theoretical large potentialities, appears limited both by their technical nature and the models developed. An exhaustive review of RS applications in human health highlights the real implication thus far regarding the diversity and range of health issues, remotely sensed data, processes and interpretations. The place of RS is far under its expected potential, revealing fundamental barriers in its implementation for health applications. The selection of images is done by practical considerations as trivial as price and availability, which are often not relevant to addressing health questions requiring suitable resolutions and spatio-temporal range. The relationships of environmental variables from RS, geospatial data from other sources for health investigations are poorly addressed and usually simplified. A discussion covering the potential of RS for human health is developed here to assist health scientists deal with spatial and temporal dynamics of health, by finding the most relevant data and analysis procedures. PMID- 16044677 TI - Application of Geographical Information Systems and Remote Sensing technologies for assessing and monitoring malaria risk. AB - Despite over 30 years of scientific research, algorithm development and multitudes of publications relating Remote Sensing (RS) information with the spatial and temporal distribution of malaria, it is only in recent years that operational products have been adopted by malaria control decision-makers. The time is ripe for the wealth of research knowledge and products from developed countries be made available to the decision-makers in malarious regions of the globe where this information is urgently needed. This paper reviews the capability of RS to provide useful information for operational malaria early warning systems. It also reviews the requirements for monitoring the major components influencing emergence of malaria and provides examples of applications that have been made. Discussion of the issues that have impeded implementation on a global scale and how those barriers are disappearing with recent economic, technological and political developments are explored; and help pave the way for implementation of an integrated Malaria Early Warning System framework using RS technologies. PMID- 16044678 TI - Geographical Information Systems in parasitology: a review of potential applications using the example of animal trypanosomosis in West Africa. AB - The epidemiology of vector-borne diseases is complex due to the variability in the ecology of the different actors involved, i.e. hosts, parasites and vectors. The transmission of African animal trypanosomosis in the West-African savannah region is an excellent example of this complexity: riverine tsetse flies have an heterogeneous distribution along the rivers, depending of suitable habitats, and transmit pathogenic trypanosomes were they use domestic animal as feeding hosts. Contrasting epidemiological situations may thus occur at the local scale, and a broad view of the overall environment is necessary to quantify the interfaces in time and space between hosts and vectors. Geographical Information Systems (GIS) can provide new insight into the study of such complex epidemiological processes. GIS is a powerful technology that has been used mainly in map-making, and an enormous amount of knowledge can be gained simply by geographical data projection. GIS also allows juxtaposition of different types of information, creation of new variables, testing of theories and correlation, and generating of predictive models. The purpose of the present paper is to exemplify the potential application of GIS using a recent study carried out on animal trypanosomosis in a cattle-raising area of Burkina Faso. PMID- 16044679 TI - A study of the environmental determinants of malaria and schistosomiasis in the Philippines using Remote Sensing and Geographic Information Systems. AB - Malaria and schistosomiasis are two water-related parasitic diseases affecting millions of people worldwide particularly tropical and subtropical countries. In the Philippines, malaria is found in 72 out of 78 provinces while schistosomiasis is endemic in 24 provinces. The Anopheles mosquito and the Oncomelania snail involved in the transmission of these diseases depend on certain environmental determinants that support mosquito and snail populations. This study, done for the first time in the Philippines, successfully showed how Remote Sensing (RS) and Geographical Information Systems (GIS) can be effectively used in showing how these environmental factors affect the spatial distribution of these two diseases. The study sites, i.e. the municipalities of Asuncion and Kapalong, are known endemic sites for both malaria and schistosomiasis. Georeferenced data enabled visualization of prevalence data in relation to physical maps thus facilitating assessment of disease situation in the two municipalities. RS and GIS data proved that other factors aside from climate influence the epidemiology of the diseases in the two sites. Topography and slope as main physical factors influence the vegetation cover, land use and soil type prevailing in particular areas. In addition, water sources especially irrigation networks differed in various places in the study sites in turn affecting the magnitude and distribution of malaria and schistosomiasis. Significant correlations found between the diseases and the environmental variables formed the basis for development of models to predict the disease prevalence in the two municipalities. Proximity to snail breeding sites and irrigation networks and the highly agricultural nature of the barangays were identified as the most common factors that define the high prevalence areas for schistosomiasis confirming the fact that conditions that support the snail populations will in turn favor the presence of the disease. For malaria, the predictive models included temperature, humidity, soil type, predominance of reproduction brush, presence of cultivated areas, distance from deep wells and distance from conventional water source which are in turn influenced by the factor of elevation. PMID- 16044680 TI - Modeling the distribution of Schistosoma mansoni and host snails in Uganda using satellite sensor data and Geographical Information Systems. AB - The potential value of MODIS satellite sensor data on Normalized Difference Vegetation Index (NDVI) and land surface temperatures (LST) for describing the distribution of the Schistosoma mansoni-"Biomphalaria pfeifferi"/Biomphalaria sudanica parasite-snail system in inland Uganda, were tested by developing annual and seasonal composite models, and iteratively analysing for their relationship with parasite and snail distribution. The dry season composite model predicted an endemic area that produced the best fit with the distribution of schools with > or =5% prevalence. NDVI values of 151-174, day temperatures of 26-36 degrees C, and night temperatures of 15-20 degrees C were used as criteria for the prediction model. Using the same approach with host snail data indicated that most of Uganda is suitable "B. pfeifferi"/B. sudanica habitat, except for possibly the north-eastern region of the country. The parasite, however, appears to be restricted in its distribution in both the north-eastern and the south western regions of Uganda. The absence of disease in the south-west can not be attributed to the absence of snail hosts. Results suggest a combination of satellite sensor data on temperature and standard climate data on precipitation, as the best ecological determinants of the S. mansoni-"B. pfeifferi"/B. sudanica system. Satellite composite models and logistic regression analysis, suggest low night time temperature as one of the significant factors inhibiting S. mansoni transmission in the south-western highland areas of Uganda. The developed models are, however, unique, representing species-specific ecologic preferences of the S. mansoni-"B. Pfeifferi"/B. sudanica system in inland Uganda. Further validation studies are needed to test the value of the model in other countries in East Africa. PMID- 16044681 TI - A potential impact of climate change and water resource development on the transmission of Schistosoma japonicum in China. AB - There is growing consensus among climate modellers that the unusual global warming observed in the last decades of the 20th century is primarily forced by human activities, namely greenhouse gas increases in the atmosphere. Global warming will trigger alterations in physical and biological systems, including shifts in the spatio-temporal distribution of disease vectors, but the nature and extent of these changes are poorly understood. The purpose of the present study was to assess the potential impact of climate change and water resource development on the distribution of Oncomelania hupensis, the intermediate host snail of Schistosoma japonicum. We employed two 30-year composite datasets comprising average monthly temperatures collected at 623 observing stations throughout China, spanning the periods 1961-1990 and 1971-2000. Temperature changes were assessed spatially between the 1960s and 1990s for January, as this is the critical month for survival of O. hupensis. Our database shows that January temperatures increased at 590 stations (94.7%), and that China's average January temperature in the 1990s was 0.96 degrees C higher than 30 years earlier. The historical 0-1 degrees C January isotherm, which was considered the approximate northern limit of S. japonicum transmission, has shifted from 33 degrees 15' N to 33 degrees 41' N, expanding the potential transmission area by 41,335 km2. This translates to an additional 20.7 million people at risk of schistosomiasis. Two lakes are located in this new transmission area that form part of the proposed South-North water transfer project. Climate change, coupled with water resource developments in China, may pose additional challenges for the control of schistosomiasis. PMID- 16044682 TI - Use of Geographic Information Systems in the development of prediction models for onchocerciasis control in Ethiopia. AB - A risk assessment model was developed for onchocerciasis distribution and its control in Ethiopia using Geographic Information System (GIS) methods. GIS data analysis was done to generate 3 separate risk models using selected environmental features of (1) earth observing satellite data on Normalized Difference Vegetation Index (NDVI) and midday Land Surface Temperature (LST) prepared from AVHRR sensor data of the Global land 1-km project for the years 1992 and 1995, (2) FAO agroclimatic databases from the Crop Production System Zone (CPSZ) of the Intergovernmental Authority on Development (IGAD) sub-region of East Africa, and (3) a climate-based forecast index based on the growing degree days (GDD) and water budget concepts. Parasitological data used for the analysis were published and unpublished reports of onchocerciasis surveillance made between 1969 and 2000 in various parts of the country. Analysis of queries based on 1992 and 1995 annual wet and dry season data of the Global land 1-km project resulted in a predictive value of 95.1%, 94.0% and 96.3%, respectively, using data values extracted from buffers centered on sites above 5% prevalence. The model based on CPSZ data predicted an endemic area that best fit the distribution of sites over 5% prevalence; the query was based on CPSZ values of average altitude (442-2134 m), annual mean temperature (18-28 degrees C), annual rainfall (822-1980 mm), annual potential evapotranspiration (1264-1938 mm), rain minus potential evapotranspiration (-124 - 792 mm), average NDVI x 100 (2000-5000) and average terrain percent slope (9-34). The climate-based model based on GDD and water budget predicted high risk to severe risk areas in the western and southwestern parts of the country. All three of the models predicted suitable areas for the transmission of onchocerciasis outside known endemic areas, suggesting the need for ground-based validation and potential application to current community directed treatment programs with ivermectin (CDTI) for control of onchocerciasis in Ethiopia. PMID- 16044683 TI - Geographical patterns of onchocerciasis in southern Venezuela: relationships between environment and infection prevalence. AB - Onchocerciasis is a chronic filarial infection transmitted by Simulium flies that has a focal geographical distribution in Latin America. The southern Venezuelan focus has a gradient of endemicity that includes the largest number of hyperendemic communities in the continent, many of them in remote forest and mountainous areas, where it is an important public health problem among the Yanomami indigenous population. The recent introduction of Geographical Information Systems (GIS) tools and a landscape epidemiology approach for study of vector borne diseases is helping to understand relationships between environment and transmission dynamics of onchocerciasis. Striking differences in the transmission dynamics of onchocerciasis between different river courses were detected. A significant relationship between onchocerciasis and temperature was also demonstrated. The geologic substrate, kind of landscape and vegetation seemed also to influence the transmission of onchocerciasis. In the Venezuelan Amazon, different kinds of landscapes associated with distinctive vector species, show different intensities of transmission of onchocerciasis. In this sense, landscape analysis aided by GIS, may prove to be a useful tool for better identification of the spatial distribution of onchocerciasis risk in the Orinoco basin. PMID- 16044684 TI - Geographical Information Systems risk assessment models for zoonotic fascioliasis in the South American Andes region. AB - The WHO recognises Fasciola hepatica to be an important human health problem. The Andean countries of Peru, Bolivia and Chile are those most severely affected by this distomatosis, though areas of Ecuador, Colombia and Venezuela are also affected. As part of a multidisciplinary project, we present results of use of a Geographical Information Systems (GIS) forecast model to conduct an epidemiological analysis of human and animal fasciolosis in the central part of the Andes mountains. The GIS approach enabled us to develop a spatial and temporal epidemiological model to map the disease in the areas studied and to classify transmission risk into low, moderate and high risk areas so that areas requiring the implementation of control activities can be identified. Current results are available on a local scale for: (1) the northern Bolivian Altiplano, (2) Puno in the Peruvian Altiplano, (3) the Cajamarca and Mantaro Peruvian valleys, and (4) the Ecuadorian provinces of Azuay, Cotopaxi and Imbabura. Analysis of results demonstrated the validity of a forecast model that combines use of climatic data to calculate of forecast indices with remote sensing data, through the classification of Normalized Difference Vegetation Index (NDVI) maps. PMID- 16044685 TI - Statistical modelling of the spatial distribution of prevalence of Calicophoron daubneyi infection in sheep from central Italy. AB - Statistical modelling for Disease Mapping and Ecological Analysis is of particular importance in veterinary parasitology because environmental characteristics can affect parasite distribution. However, the main difficulties relate to the concentration of animal populations within farms, which contrasts to the study of wild animal populations. In the present paper we report the results of a cross-sectional coprological survey designed to study the presence and distribution of the rumen fluke Calicophoron daubneyi--which causes paramphistomosis, a snail borne disease--in pastured sheep living in the Latina province of central Italy. We show how techniques derived from human epidemiology can be used to study the spatial distribution of parasite infection in animals. We proposed a hierarchical Bayesian model with random terms for unstructured variability (heterogeneity) to account for local farm characteristics and spatially structure terms (clustering) to cope with medium-large scale environmental characteristics. PMID- 16044686 TI - Remote Sensing and Geographic Information Systems and risk of American visceral leishmaniasis in Bahia, Brazil. AB - The spatial distribution of American visceral leishmaniasis (VL) was studied within the context of the environmental characteristics of northwest Bahia State in Brazil during an epidemic year. Geographic Information Systems (GIS) and Remote Sensing (RS) were used to characterize the landscape epidemiology of VL in order to identify and map high risk areas and endemic zones in a northwestern Bahia study area. Normalized Difference Vegetation Index (NDVI) was shown to be one of the most important risk factors in the area of study. Low NDVI values were related to high numbers of sand flies and high numbers of human and canine VL positive cases. Caatinga vegetation type was the dominant vegetation type in the endemic area. The use of RS and GIS allowed the identification of classes of VL risk that may be useful information to guide control program interventions. PMID- 16044687 TI - Geographical Information Systems and on-line GIServices for health data sharing and management. AB - Integrating Geographical Information Systems (GIS) technology and public health experience may represent a solution for a better comprehension of spatial and temporal trends of phenomena. Useful applications can be built that support practitioners in their daily tasks, from risk assessment to prevention programmes. Also, making available data on the Internet through GIServices represents an important goal. Institutions and public health practitioners may benefit from the technological integration of GIS, the Web, handheld and mobile global positioning systems (GPS) devices. Expert users may be supported in deriving thematic maps which represent a spatial synthesis documentation starting from an analytic study expressed in terms of numbers and features. In this paper we show an example of an on-line data sharing and processing application, emphasizing ways GIS can provide added value to health research and management. PMID- 16044688 TI - A classification model of grazing areas in southern Italy using Remote Sensing data. AB - Detection of vegetation typologies is of particular interest in epidemiological studies on animal diseases. This paper is focused on the automatic detection of grazing areas in southern Italy browsed by equines, oxen and sheep, mostly sedentary, with many in fenced areas for overnight stays. Results revealed Satellite Remote Sensing was an indispensable tool in area surveys for vegetation cover characterization. Our classification model shows an accuracy level of 90.21% and a precision of 92.69%. Results suggest similar application protocols can be used in areas with different vegetation cover to characterizing potential infection risk areas for geohelminths and other diseases. PMID- 16044689 TI - Notes on cystic echinococcosis in the Mediterranean. AB - Cystic Echinococcosis (CE), caused by Echinococcus granulosus, is present from the beginning of history and in the Mediterranean it is linked to the dog-sheep cycle. The Mediterranean area possesses many features favouring CE. Positive and negative influences derived from the action of the European Community and from recent developments. The control measures of CE have political, economic, public health and environmental implications. Dog population and dog-transmitted zoonoses control, improvement of slaughtering procedures and the destruction of infected viscera, health education, interprofessional cooperation are able individually to constitute a contraposition to CE and combined to compose a control program. Epidemiological surveillance and control of CE in the Mediterranean are coordinated by the WHO Mediterranean Zoonoses Control Centre of Athens. PMID- 16044690 TI - Socio-economic impact of cystic echinococcosis and of its control: some data and considerations. AB - The socio-economic impact of cystic echinococcosis (CE), caused by Echinococcus granulosus, is reviewed with special reference to the following topics: consequences in man and livestock, costs and benefits of control programmes and economic procedures for evaluating control programmes. Examples of some important costs and benefits are given. Many consequences in man and livestock are difficult to evaluate from an economic point of view, because some basic data are difficult to obtain in many countries. However, the socio-economic evaluation of the consequences of CE and of the present and future control actions proves indispensable to best use available resources and possibly tailor control stategies. PMID- 16044691 TI - Natural history of cystic echinococcosis in humans. AB - This study provides a contribution to understanding of the natural history of the hydatid cyst during its evolution in the human liver examining the high morphostructural variability of larval forms of Echinococcus granulosus. A detailed study of a large sample of intact cysts removed from patients surgically treated by means of total pericystectomy, has enabled to outline the different developmental stages of the parasite over time, up to its death and complete degeneration. PMID- 16044692 TI - Twenty years of percutaneous treatments for cystic echinococcosis: a preliminary assessment of their use and safety. AB - Image-guided percutaneous treatments for echinococcal cysts were introduced in the mid-eighties. Today they represent a third therapeutic option, after surgery and benzimidazole derivatives. Two types of percutaneous treatments are available, based on the destruction of the germinal layer or the evacuation of the endocyst. To assess the extent of their use and their safety, a Medline search of the literature on this subject was performed. The number of cysts treated, their anatomical sites, the complications and, length of follow-up (when available), were all examined. The results show that percutaneous treatments for cystic echinococcosis are safe and efficacious in selected anatomical sites, provided basic safety issues are correctly addressed. However, before drawing final conclusions, a more detailed analysis of the literature is needed. Percutaneous treatments could be simplified and made more effective if a scolecidal agent could be found that melts the entire endocyst without causing harm to the biliary epithelium. PMID- 16044693 TI - Epidemiology of hydatidosis in the province of Sassari, Italy. AB - Cystic echinococcosis is endemic in certain parts of the world, including Sardinia, Italy. It was performed a study in the province of Sassari in order to evaluate the incidence of the infection in man and the effects of control programs since 1964 to 2002. Data obtained by surgical records, hospital discharge forms, radiological and pathological files were collected using a case report form. During the years 1964-2002, 2702 new cases were identified (average annual incidence: 17 per 100,000) and 1981 (73.3%) were submitted to surgical treatment. In 57.3% municipalities no cases were observed during the years 1998 2002. Males are more affected (56.2%), mostly farmers-shepherdess (68.6 per 100,000) and pensioners (59.6 per 100,000). Control measures led to a significant decline in the incidence rate of hydatidosis during the period 1964-2002, dropping by 27.6 per 100,000. The mean age of surgical patients increased during the years of surveillance, such as the surgical liver/lung ratio as a consequence of a cohort effect. The durability of control programs is the corner stone for obtaining a significant decrease of this infection. PMID- 16044694 TI - Immunobiology of cystic echinococcosis. AB - The present report focuses on the ability of cystic echinoccocosis metacestode to survive for a long time, despite host immunity, by developing avoidance strategies. The tactics believed to come into play, ranging from intrinsic parasitic factors to host-related mechanisms, are briefly discussed and the importance of studies on experimental models is highlighted particularly in relation to furthering the theoretical understanding of the dynamic equilibrium between host and parasite, and to the feasibility of practical approaches in controlling the disease by artificial manipulation of the immunoregulatory mechanisms. PMID- 16044695 TI - Veterinary public health activities at FAO: echinococcosis/hydatid disease. AB - Cystic hydatidosis is a zoonotic disease that remain as a significant cause of human morbidity and mortality in many parts of the world. The disease has veterinary public health implications. FAO is involved with some activities in the control of echinococcosis/hydatid disease: within the Animal Production and Health Division the Veterinary Public Health (VHP) Programme is constituted by members of the different Services (Animal Health, Animal Production, and Livestock Policy) within the Division. FAO regular programme has also established a global network of professionals directly involved in VPH. Furthermore FAO's Technical Cooperation Projects (TCP) is a tool to assist member countries in responding to urgent and unforeseen demands. PMID- 16044696 TI - Human cystic echinococcosis in Sardinia during the 20th century. AB - At the beginning of the 20th century in Sardinia human Cystic Echinococcosis was considered a highly endemic disease by all the health operators. In the early twenties the number of reported cases was 429, but incidence appeared rising due to increase in availability of data and to improvements in diagnosis. The fall of personal and public hygienic conditions together with the drift from urban to rural areas during the second world war, had in fact facilitated the possibilities of infection, causing a strong growth of surgical cases during the post war years, so that incidence rate remains high and almost unvaried until the end of the eighties. Comparative analysis of young and probably old cysts indicates that this high rate of surgical cases was attributable to treated hepatic cysts in elderly. On the contrary the continuous reduction in the number of young cysts clearly shows a slow but constant decrease of the infection rate. PMID- 16044697 TI - Cystic echinococcosis in Italy from the 1950s to present. AB - In Italy the epidemiological pattern of cistic echinococcosis (CE) is incomplete and the information for most regions is out of date, contradictory, and almost exclusively limited to the intermediate hosts. The disease is found most frequently in particular social and economic conditions: widespread use of extensive or semi-extensive sheep farming, illegal slaughtering, and high numbers of sheepdogs and other types of dogs. The highest incidence in sheep is found in Sardinia (70.6-92.8%), Sicily (6.5-36.5%), Basilicata (5-28%), Abruzzo (22%) and Tuscany (47%). It North Italy, it is never higher than 0.5% in slaughtered sheep. No data are available on the biomolecular characterization of the strains of E. granulosus in Italy, apart from Sardinia and recently Lazio. G1 (Sheep strain), G7 (Pig strain) G2 (Tasmanian sheep strain) have been identified in Sardinia and G1 and G3 (Buffalo strain) have been recently isolated in Lazio. In Italy, CE has was also found in buffaloes (2.63-9.8%) and horses (<1%). However, further epidemiological surveys and genotyping study are necessary. The small quantity of up to date information on the diffusion of E. granulosus in dogs (Abruzzo 4%, Sardinia 6-10% and Sicily 19.3%) highlights the need for modern, fast, sensitive and low risk diagnostic methods which would provide a true picture of the pattern of the infection in this host. PMID- 16044698 TI - Health education and formation: essential tools into the echinococcosis/hydatidosis prevention programs. AB - Health Education represents essential elements in the control and eradication campaigns of Echinococcosis/Hydatidosis. The basic elements and the applicatory principles and methodologies are examined. Moreover, the most appropriate intervention methodologies are defined through reference target. There is also a description of the experiences and the methodologies adopted in different Countries of the Mediterranean basin, where the disease is endemic. PMID- 16044699 TI - Cystic echinococcosis in Sardinia: the current role of sheep. AB - Cystic echinococcosis is one of the most widespread parasitosis in the Mediterranean region. Unfortunately this is also true for Sardinia for various reasons, among which is the close relationship between sheep and dogs in the farms. This work first epidemiologically analyses hydatidosis in sheep in Sardinia and then examines the most important causes of the persistence of these metacestodosis in sheep. The work looks at the factors which are responsible for the enormous quantity of illegal slaughtering and the widespread habit of abandoning the sheep carcasses in the grazing areas, as well as possible initiatives which may eliminate these practices. Such initiatives must include reducing the financial costs and bureaucracy involved when sheep are slaughtered in abattoirs, and also increasing the value of sheep meat. In conclusion new checks and controls are hoped for at a political level which will increase the financial support for the farmers and encourage the use of recombinant vaccines which have already been tested in Sardinia. These are extremely useful for effectively curbing cystic echinococcosis. PMID- 16044700 TI - Critical points in the immunodiagnosis of cystic echinococcosis in humans. AB - This study discusses the immunodiagnosis of cystic echinococcosis (ce, caused by Echinococcus granulosus). The detection by immunoblotting of antibodies specific for the 8 kDa subunit of antigen B and in particular the IgG4 subclass expression, seems the most promising serodiagnostic tool. Despite the development of molecular methods, nowadays there is no standard, highly sensitive, and specific test available for antibody detection in CE. Furthermore, because serological tests can give only a limited support to clinical findings there is a clear need for new advances in immunodiagnosis of E. granulosus infection. PMID- 16044701 TI - An up-to-date on clinical management of human cystic echinococcosis. AB - More than one thousand of patients with more than two thousand of Echinococcus granulosus hydatid cysts were observed during the last 20 years. The therapeutic approach ranged from chemotherapy with benzimidazole carbamates (mebendazole and albendazole) to surgery, percutaneous non conventional treatment, and a "wait and see" approach, on the basis of the clinical picture, of the compliance and of the socio-economic background of each patient. The observed results, mainly those regarding chemotherapy, were briefly summarized and discussed. Most important point to underline is the need of data about a more prolonged follow-up on wider casuistries of patients both treated (with chemotherapy, with surgery, with percutaneous non conventional treatment) and not treated, to obtain a more suitable evaluation of the long-term outcome of the performed clinical approach. Moreover, a multidisciplinary approach is advisable to assure a better clinical management to patients with cystic echinococcosis. PMID- 16044702 TI - The diagnosis of Echinococcus granulosus in dogs. AB - The problem of diagnosing Echinococcus granulosus in dogs has still only been partially resolved, even after the advent of biotechnology. The eggs of taeniid Cestoda are extremely similar, and thus identification by microscopic examination of the faeces is risky and non-specific. For this reason, Echinococcus granulosus was traditionally diagnosed in dogs ante mortem after an arecoline hydrobromate purge. The faeces were examined macro and microscopically to establish if the adult tapeworm or its proglottids were present. Although this method is 100% specific, it is bio-hazardous and time-consuming, requires trained personnel, and its sensitivity varies. In the 1990s copro-antigens were discovered and characterised. These are released by the adult worm in the faeces. This made it possible to use enzyme-linked immune-adsorbent assay (ELISA) for in vitam diagnosis of Echinococcus granulosus. In recent years several PCR protocols have been published on the identification of Echinococcus granulosus DNA from eggs or from adult parasites and new ways of diagnosing this cestode have been developed. PMID- 16044703 TI - Human echinococcosis in the Emilia-Romagna region (northern Italy) in the years 1997 to 2002: an updating. PMID- 16044704 TI - Evaluation of different diagnostic methods to detect Echinococcus multilocularis in the final host. PMID- 16044705 TI - Copro-diagnosis of Echinococcus multilocularis by a nested PCR in red foxes (Vulpes vulpes) from northern Italy. PMID- 16044706 TI - Pilot vaccination project for the control of hydatid disease in Matera province (southern Italy). PMID- 16044707 TI - An epidemiological updating on cystic echinococcosis in cattle and sheep in Sicily, Italy. PMID- 16044708 TI - Echinococcus granulosus in the wolf in Italy. AB - During the period 1987-1999, 119 wolf cadavers were examined and checked for the presence of Echinococcus granulosus. All the animals were retrieved along the whole Apennines range of distribution of the species in Italy and most of them were illegally killed. Eighteen wolves resulted positive (15%). The mean intensity was 697.5. The force of infection for prevalence was 8.2 year(-1). The prevalence of the parasite was significantly and positively influenced by the local prevalence of cystic echinococcosis (CE) in sheep. Mean intensity was significantly and positively influenced by both the age of the wolf and the prevalence of CE in sheep. A deterministic model was used in order to simulate a purely theoretical sylvatic cycle of the parasites having the wolf as the only definitive host with 15% of prevalence. The expected prevalence of CE in wild intermediate species ranges between 10% and 25%. This prevalence overlaps the one observed in sheep. Even if both the wolf and the wild ungulate populations are increasing, the wolf still acts as a part of the main dog-sheep cycle of the parasite. PMID- 16044709 TI - Casual finding of a hydatid cyst during an autopsy in Veneto region (NE Italy). PMID- 16044710 TI - An updating on the epidemiological situation of Echinococcus multilocularis in Trentino Alto Adige (northern Italy). PMID- 16044711 TI - Recombinant antigens of Echinococcus granulosus recognized by IgE and IgG4 of sera from patients with cystic echinococcosis. PMID- 16044712 TI - Distribution of hydatidosis in slaughtered animals in Umbria region from 1995 to 2004: a retrospective analysis. PMID- 16044713 TI - From Nairobi to Sassari, a realistic role for Italian hydatidology. Thoughts from the XXI International Congress of Hydatidology. PMID- 16044714 TI - Cytokine expression in the follow-up of patients with cystic echinococcosis. PMID- 16044715 TI - An epidemiological and biomolecular survey of cystic echinococcosis in cattle in Sardinia. PMID- 16044716 TI - Preliminary data on Echinococcus granulosus (Batsch, 1786) in dogs from Lombardia and Marche regions (northern and central Italy). PMID- 16044717 TI - Cystic echinococcosis in the Campania region (southern Italy). PMID- 16044718 TI - [Nurses are aware of the future]. PMID- 16044719 TI - [Europe searches for its regulations. Ambulatory nurses bring new problems to the Common Market development and the liberalization of the service sector]. PMID- 16044720 TI - [Psychiatric nursing in institutions. Changes in guidelines in the regulation of psychiatric nurses in institutions make staff psychiatrists more susceptible to regulation]. PMID- 16044721 TI - [Paper is patient.. Nursing process and nurse documentation of the practicing bureaucracy is never ending. The medical profession is the most the sharply critical of patient care]. PMID- 16044722 TI - [Dosage is not simply dosage--the correct aid helps to make medication dispensing systems assist the ambulatory nurse as well the staff nurse in giving the correct medication dosage]. PMID- 16044723 TI - [Health promotion in education and continuing education in nursing. The project of transfer-oriented training in technical schools will further the development of the health promotion concept and its curricular implementation]. PMID- 16044724 TI - [Nursing and midwifery: pioneering in a new field. The new continuing education course for nurses and midwives "Family health for nurses and midwives prevention and health promotion" will start in October 2005. ]. PMID- 16044725 TI - [Social partnerships in nursing. In Hesse there is a development of nursing standards for productive cooperation for patient or residents of nursing institutions. From the social partnership model of 2001 which has become involved with the "Social partnership of Shwalm-Eder:]. PMID- 16044726 TI - [Nursing for the chronically ill. Not treatment-oriented medicine, but special care concepts of health nursing will be asked in the future]. PMID- 16044727 TI - [Interdisciplinary nursing trends. Position paper on the cooperation between clinical social work and nursing]. PMID- 16044729 TI - [Forum on the direction for wards 2005: conduct also takes courage]. PMID- 16044728 TI - [Trusting is good...but control is better: difference of opinion in Innsbruck concerning hand or shoe perforation and the consequences]. PMID- 16044730 TI - [Nurses need not worry about their workplace. Northwest Germany features the "food dispute"]. PMID- 16044731 TI - [Treatment comes from the correct tactic: where one can speak freely concerning the future about the unusual case. Therefore, one must be prepared with the proper tactic]. PMID- 16044732 TI - [Concerning health of Nurses, an expert gives advice about how even nurses can stay in top physical condition]. PMID- 16044733 TI - [Sexual abuse, Equality, whether it exists among colleagues or patients: no one in nursing should allow sexual abuse. How to achieves this and which precautions to take]. PMID- 16044734 TI - [Do we need a room for nurses?]. PMID- 16044735 TI - Chronic suppurative otitis media and related complications at the University Clinic of Kinshasa. AB - During the five-year period from 1998 to 2002, the ENT department of the University Clinic of Kinshasa treated 343 patients suffering from chronic suppurative otitis media (CSOM) and complications. Two hundred and seven patients had simple, 28 had cholesteatomatous form of CSOM, and 108 patients presented with intratemporal or intracranial complications. Roughly half of the patient group consisted of children younger than 10 years. The simple form of CSOM affects men and women equally, but the cholesteatomatous form was more prevalent among men. Otorrhoea and hearing loss were the major presenting symptoms. Hearing loss was moderate to severe in 2/3 and slight in 1/3 of the audiometrically tested ears. A mastoidectomy was performed on 55% of the patients. Roughly 1/3 of the patients presented with complications, the majority of which (69) were subperiostal abscesses. Complications other than subperiostal abscesses were associated with a high morbitidy and a few patients even died. Our data suggest that the prevalence of CSOM in the Kinshasa area is at least 4% and, in fact, has not declined over the last 20 years. PMID- 16044736 TI - Warthin's tumour and smoking. AB - OBJECTIVE: In an evaluation of our patients with parotid gland neoplasms, we noticed that patients with a Warthin's tumour were heavy smokers. The aim of this study was to confirm earlier findings in the literature concerning a possible association between smoking and the development of a Warthin's tumour. METHODS: A case control study was performed using the clinical records and discharge letters of all consecutive patients with a Warthin's tumour in the pathology database of our hospital covering the last 15 years. Patients with a pleomorphic adenoma and a group of patients visiting our audiology department were used as controls. RESULTS: A smoking history was found in 97.5% of the patients with a Warthin's tumour. Of the patients with a pleomorphic adenoma, 59% had a smoking history; 56.5% of the audiology group had a smoking history. Mean age at the time of the operation was 60.1 years of age in the Warthin's tumour group and 48.6 for the pleomorphic adenoma group. CONCLUSION: The mean age for the development of a Warthin's tumour is ten years older than for a pleomorphic adenoma. Furthermore, the development seems to be closely related to smoking habits. PMID- 16044737 TI - Completion thyroidectomy after the unexpected diagnosis of thyroid cancer. AB - The optimal surgical management of well-differentiated thyroid cancer (DTC) remains a controversial topic. Preoperative and peroperative investigations quite frequently fail to detect thyroid cancer in cold nodules, and only postoperative histological examination reveals malignancy. In these cases many physicians perform a completion thyroidectomy. Others recommend a conservative approach with close follow-up because of the increased risk of complications after re operation. In our department, routine management includes completion thyroidectomy once the histopathological report concludes that there is carcinoma, except in cases of papillary carcinoma measuring less than 1 cm. The aim of our study was to determine the incidence of contralateral malignancy and of complications after completion thyroidectomy. We reviewed the records of 29 patients--25 women and 4 men-- who all underwent completion thyroidectomy because of an unexpected diagnosis of DTC. Residual malignancy was found in 12 patients (41.4%) after completion thyroidectomy. In ten patients (34.5%) the malignancy was localised in the contralateral lobe and two patients (6.9%) had lymph node metastases. Postoperative transient hypocalcaemia (< 8.0 mg/dl) occurred in five patients (17.2%) and permanent hypocalcaemia (lasting longer than 6 months) was a feature in two patients. One patient suffered transient laryngeal nerve injury occurred in one patient and there were no permanent lesions. In conclusion, we found residual DTC in 41.4% of patients undergoing reintervention. Because of the rather low re-operation rate, we prefer to perform a completion thyroidectomy to remove potential occult malignancy and to allow for postoperative 131I-treatment in all patients with a diagnosis of malignancy in their thyroid lobectomy specimen, with the exception of papillary carcinoma < 1 cm. PMID- 16044738 TI - Eosinophilic fungal rhinosinusitis (EFRS): a distinct CT/MRI-entity? A European experience. AB - OBJECTIVE: To determine the value of radiological features in the diagnosis of Eosinophilic Fungal Rhinosinusitis (EFRS). STUDY DESIGN: Retrospective review of the radiological materials of 65 patients with documented Eosinophilic Fungal Rhinosinusitis treated at the same institution. METHODS: Evaluation by the ENT surgeon and the head and neck radiologist. RESULTS: EFRS was more common in female patients in this series. Fifty-four (83%) patients were above 30 years of age, with a peak of 18 patients (27.7%) in the seventh decade. All the patients except one (98.5%) showed bilateral mucosal thickening on unenhanced CT scans. Thirty-eight patients (58%) showed increased intrasinus attenuation on unenhanced CT scans. Thirty-seven patients (57%) showed opacification of at least one sinus; 25 (38%) showed osteitis; 11 (17%) had erosion of the sinus wall and only one patient showed minor expansion of an involved sinus. In 6 patients, typical hyperattenuation patterns on CT scans, together with distinctive MRI images, were highly suggestive of EFRS. CONCLUSION: Our data show that hyperattenuation on CT images with bone window settings suggests the presence of EFRS. This hyperattenuation is more clearly seen with soft-tissue window settings. When necessary, adjunctive MRI can provide information which might be highly predictive for the diagnosis of EFRS. However, non-specific imaging findings of chronic rhinosinusitis (CRS) should also be seen as possible EFRS pathology. PMID- 16044739 TI - Fungus-like sinusitis. AB - Fungus-like sinusitis is also called eosinophilic mucin rhinosinusitis (EMRS). This terminology is purely descriptive and refers to a rhinosinusitis, whose clinical presentation and imaging are similar to those of a noninvasive fungal sinusitis but in which fungus could not be demonstrated pathologically using the conventional silver staining technique or on culture. The authors report 2 clinical cases and recall the diagnostic criteria for the different forms of fungal sinusitis in immunocompetent patients. PMID- 16044740 TI - Pneumosinus dilatans and orbital meningioma in neurofibromatosis type 2. AB - We report on a 9-year-old boy with neurofibromatosis type 2 and a progressive proptosis of the left eye. Imaging studies revealed a pneumosinus dilatans of the left ethmoidal and frontal sinuses and an adjacent left orbital mass. Incision biopsy of the orbital mass showed a meningiothelial meningioma. Orbital and intracranial meningiomas are frequently associated with pneumosinus dilatans. This report illustrates the relationship of pneumosinus dilatans, intra-orbital meningioma and neurofibromatosis type 2. PMID- 16044741 TI - Blunt external trauma to the trachea: report of two cases. AB - We present two cases of blunt external trauma to the trachea. The first was a 19 year-old man who had suffered rupture of the first tracheal ring as a consequence of attempting suicide by hanging from a cable. The second was an 11-year-old boy who fell from a bicycle and struck his neck against the handlebars. Fracture of the second tracheal ring was found at surgery. Both cases were treated by primary repair. A surgical exploration is essential in the case of subcutaneous emphysema, even if no respiratory distress is present and a radiological evaluation is negative. Follow up, according to the extent and type of injury, is recommended to prevent post traumatic stenosis of the trachea. PMID- 16044742 TI - Laryngeal paraganglioma: an endoscopic diode-laser-assisted surgical approach: a case report. AB - Laryngeal paraganglioma is a rare vascular neuroendocrine benign tumour. Surgery is the standard treatment. The pre-operative gold standard is Magnetic Resonance Imaging with Gadolinum contrast. Computed tomography scans and angiography are also useful in making the diagnosis of paraganglioma before surgical intervention. A case of a large supraglottic paraganglioma is described. Complete excision of the tumour was achieved using microlaryngoscopical-laser surgical resection. No severe complications were observed. The patient was discharged from hospital without any complaints on the fourth post-operative day and she was disease-free 36 months after surgery. The effectiveness of this conservative endoscopic surgical procedure compared to the external approaches is discussed and it can be concluded that the endoscopic diode-laser resection of even extended laryngeal paraganglioma provides a good operative field and is a safe and effective technique. PMID- 16044743 TI - Papillary thyroid carcinoma presenting as a giant cervical mass: a case report. AB - Papillary thyroid carcinoma (PTC) is a relatively uncommon malignancy. The prognosis is generally good and the mortality rate is low. PTC is more common in younger patients; incidence is two to three times higher in women and in individuals with radiation exposure to the neck. We report on a 75-year-old woman with PTC admitted to our clinic for a giant mass in the neck dating back ten years. A hyperintense huge cystic lesion and hypo-intense central solid component were seen on T1- and T2-weighted magnetic resonance imaging. No distant metastasis was present. The PTC was treated surgically and radioiodine (131I) treatment was applied postoperatively. No recurrence was observed one year later. PMID- 16044744 TI - [Guidelines for the organization and management of disinfection and disinfestation from rodents and mosquitoes]. AB - Two recent legislative acts (D.P.C.M. 29.11.2001 and D.G.R. Veneto n. 2093, issued 02.08.2002) give to Local Health Authorities responsibility over surveillance of disinfestations activities. Authority over the implementation of these actions remains with the municipalities. Another legislative act (D.G.R. Veneto 3015) established the creation of a task force in order to define guidelines regarding new managerial methods toward disinfections, disinfestations and rodent control for public administration. The "Guidelines for the organization and management of disinfection and of rodent and mosquito control" redefine the roles for the public administration units involved, propose a new organizational model and an innovative managerial approach to activities, provides technical recommendations for the preparation of specific contracts and for the implementation of disinfestation actions. Finally, the guidelines include a proposal for the development of regulations supporting the technical propositions. PMID- 16044745 TI - Unanswered questions. PMID- 16044746 TI - Growth and development in the pediatric patient. AB - What is the most important aspect of children's dentistry? Some may argue that it is the restoration orprevention of decay and disease. Others may describe nutrition. The diagnosis and treatment of proper growth and development, I feel is the most important responsibility of our practices. What we do or don't do can affect the child for life. Unfortunately, many of us lack the abilities we need to affectively monitor or augment, if necessary, proper growth and development. The purpose of this paper is to provide us with some basic knowledge, tools and treatments. PMID- 16044747 TI - Sleep disordered breathing: orthodontics and sleep disorders dentistry. PMID- 16044748 TI - Changes in the soft tissue facial profile following orthodontic extractions: a geometric morphometric study. AB - OBJECTIVES: The aim of this study is to test the null hypothesis that changes in soft tissue facial profile in children who undergo orthodontic treatment, using either an extraction or non-extraction technique, do not differ. DESIGN: Longitudinal, retrospective. SETTING AND SAMPLE POPULATION: San Juan, PR and Aberdeen, UK. Fifty-eight consecutively treated children using fixed appliances with a mean age of 13.2 (+/- 2.1) years. EXPERIMENTAL VARIABLE: Extraction of the upper and lower, first or second premolars. OUTCOME MEASURES: The mean pre- and post-treatment, extraction and non-extraction configurations were compared using finite-element scaling analysis (FESA), incorporating a thin-plate spline interpolation. RESULTS: Post-treatment the two groups differed statistically in the premaxillary region (p < 0.05) with the non-extraction group being relatively larger in that region by 25%. For the non-extraction group after treatment, localized increases in relative size in the naso-maxillary region size of 25% (p < 0.01) were present. For the extraction group after treatment, a non-significant reduction in relative size of 15% was localized in the putative bicuspid area. CONCLUSION: Soft tissue facial differences in non-extraction and extraction cases prior to treatment may be accentuated following orthodontic treatments that elect non-extraction or extraction protocols. Robust diagnosis and treatment planning, incorporating the judicious use of extractions if appropriate, may yield optimal orthodontic outcomes. PMID- 16044749 TI - [Medical ethics and the apathetic refugee children]. PMID- 16044750 TI - [Increasing number of skin cancer cases--also among the younger]. AB - The incidence of skin cancer has been increasing for several years. This is the case in malignant melanoma as well as in squamous cell carcinoma and basal cell carcinoma. Skin cancer is most common among the elderly, but is now also more frequently found in younger people. Surgery is often the treatment of choice and this is still the case in malignant melanoma and most cases of squamous cell carcinoma. For basal cell carcinoma, Bowen's disease and actinic keratoses, however, alternative treatments should be considered. Today, dermatologists can offer patients with skin cancer several new treatments besides surgery. These new treatments seem to have the same efficacy but better cosmetic results compared to earlier treatments. PMID- 16044751 TI - [Paleolithic diet and evolution medicine: the key to diseases of the western world]. PMID- 16044752 TI - [The paleolithic diet should be taken with a pinch of salt]. PMID- 16044753 TI - [Central venous catheter and malignant disease. The value of thrombosis prophylaxis is questioned now]. PMID- 16044754 TI - [Watch out for tularemia also in Southern Sweden!]. PMID- 16044755 TI - [Addison disease during pregnancy--a diagnostic dilemma. Symptoms are similar to normal pregnancy problems]. AB - The symptoms of primary adrenocortical insufficiency (Addison's disease) such as fatigue, anorexia, hypotension and hyperpigmentation are similar to those of normal pregnancy. Addison's disease is rare and the diagnosis can easily be overlooked during pregnancy. The concentration of corticosteroid-binding globulin (CBG) and cortisol in serum as well as urinary free cortisol increase 2-3 times during pregnancy. Therefore, the reference ranges for nonpregnant persons cannot be used during pregnancy. The diagnosis of Addison's disease in pregnancy should be based on analysis of p-ACTH which remains within the reference range in normal pregnancy until delivery. A case of Addison's disease diagnosed during pregnancy is presented and it illustrates the diagnostic difficulties. In patients taking oral estrogen containing contraceptives, serum levels of CBG and cortisol also increase 2-3 times, making s-cortisol values difficult to interpret. Also in these patients, analysis of p-ACTH is of value when suspecting Addison's disease. PMID- 16044756 TI - [Getrud Gussander--a female surgeon-pioneer who built her own hospital]. PMID- 16044757 TI - [But the emperor is naked]. PMID- 16044758 TI - [Non-therapeutic ventilation. Long-term consequences for relatives must be elucidated, too]. PMID- 16044759 TI - [We must learn how to be better to evaluate drug benefits and risks]. PMID- 16044760 TI - [No evidence that lowered PSA reference levels will help us detect those cases of prostatic cancer that will lead to morbidity and mortality]. PMID- 16044761 TI - [Comments on electromagnetic field therapy]. PMID- 16044762 TI - [School health services, occupational health services and the specialty]. PMID- 16044763 TI - [The producer-gas epidemic and an important hematologic discovery]. PMID- 16044764 TI - [Sleep apnea--an underdiagnosed complaint?]. PMID- 16044765 TI - [Collective subscription or no? Reflections from international medical press]. PMID- 16044766 TI - [Cervix cancer screening can be cost-effective. Combination of vaginal smears and the HPV test should be even more beneficial]. PMID- 16044767 TI - [The medically possible makes us want to go on too fast]. PMID- 16044768 TI - [Methylprednisolone in the treatment of acute spinal cord injury has become more and more questioned]. AB - Methylprednisolone (MP) has, through the results from the clinical multi-center studies National Acute Spinal Cord Injury Study II and III, during the 1990's become standard treatment in acute spinal cord injury (ASCI). Critical reappraisals of the data have later shown that analyses have been made on subgroups of the study-populations and argue that the results are based on statistical artefacts. This, combined with the failure to show improved functional recovery, puts into question earlier conclusions drawn on the efficacy of MP on ASCI. This review of the recent literature on the subject concludes that there is no scientific evidence to support MP as standard treatment in ASCI. PMID- 16044769 TI - [HeartScore, PC-based cardiovascular risk assessment. A cooperation between general practitioners and cardiologists]. PMID- 16044770 TI - [Can obesity be an infectious disease?]. AB - Obesity is generally considered as the end result of a long standing positive energy balance. Although no one disputes the laws of thermodynamics, some alternative suggestions for the etiology of obesity have been discussed. The expression "infectobesity" describes the possibility that obesity may be associated with viral infections or a particular behaviour of the intestinal bacterial mass. PMID- 16044771 TI - [Good results after twenty years of team work in the care of patients with HIV infections and AIDS]. AB - Since 1985 there has been a multidisciplinary HIV-team at the Department of Dermatology and Venereology at Sahlgrenska University Hospital. 209 HIV-positive patients have been treated at our clinic for shorter or longer periods. Our HIV team consists of two physicians, one nurse, two counsellors and one psychologist. During the years we have built a network of different medical specialists who we contact depending on the current problem. The work has changed dramatically during these twenty years. When we started there were hardly any antiretroviral drugs available, and our main tasks were to deal with the psychological stress of having a death-threatening and contagious disease and to diagnose and treat opportunistic infections. Today we have 20 different registered antiretroviral drugs, which are given in combinations. This has significantly decreased the morbidity and mortality of HIV infection. However, significant constraints to therapeutic success still exist, like toxicity and resistance. Today 81 patients are registered at our clinic. The overall CD4 status among the patients is very good and 85% of the patients who are receiving HAART have undetectable HIV-RNA. PMID- 16044772 TI - [Transient global amnesia--a benign condition which can also occur in young people]. AB - Transient global amnesia (TGA) occurs mostly in middle-aged and elderly individuals, and is generally believed to be very rare in individuals less than 40 years of age. We present three cases of TGA in young persons (16-22 years). They all had a medical history and presented symptoms fulfilling the criteria for TGA. Physical examinations and investigations were all normal. All three presented their symptoms while playing football. Reviewing the literature the suggested causes are partly different for TGA in old and young people, respectively. The present report confirms that TGA may also occur in younger individuals. We propose that single TGA is a benign condition also in younger persons and that the investigation should be similar to that of TGA in older age groups. PMID- 16044773 TI - [Positive attitude to prescription of generic preparations. Report from the Genvagen-project in western Gotaland]. PMID- 16044774 TI - [An evidence-based guide to writing grant proposals increases the chance of approval]. PMID- 16044775 TI - [Political passivity makes young physicians leave psychiatry in Gothenburg]. PMID- 16044776 TI - [More information on drug package inserts and counseling replace the warning triangle]. PMID- 16044777 TI - [An addition concerning spironolactone and eplerenone (Inspra) in heart failure]. PMID- 16044778 TI - [Open hernia surgery is qualitatively comparable and economically more beneficial]. PMID- 16044779 TI - [Some sequelae of whiplash injury]. PMID- 16044780 TI - [Time to re-evaluate the abortion law]. PMID- 16044781 TI - [Pluralism in medicine--pluralism or therapeutic evaluation?]. PMID- 16044782 TI - [Evaluation and pluralism from the viewpoint of constitutional law]. AB - The evaluation of a scientific method proceeds by means of the fitting attitude analysis. In a pluralistic system, there are different therapy concepts and therefore also different evaluation methods. The German legal system acknowledges this principle for the special therapy forms of phytotherapy, homeopathy, and anthroposophic medicine. For these fields, the golden standard is not the evidence-based medicine but, based on the principle of internal recognition, the acceptance of and response to the therapy concept. PMID- 16044783 TI - [The physician and the therapeutic decision]. AB - Science in medicine is never a self-fulfilling goal but always serves the purpose to provide the best medical care in a unique clinical situation. Coming to a rational therapeutic decision requires a diagnosis in the sense of allowing and legitimizing an action-oriented individual statement. The diagnosis, however, cannot be reduced to a process of abstraction and formalization, but it rather consists of the ability to allocate general designations of illness to an individual patient. This requires a person-oriented, not necessarily strictly formal knowledge. Inasmuch as guidelines should not become imperative instructions, the individual medical judgment should remain an indispensable focus of the identity of the medical profession. PMID- 16044784 TI - [Which therapy comparisons are fair?]. AB - Two approaches to treatment comparison are discussed: According to the principles of evidence-based medicine (EBM), methods of comparison are graded on a study type scale, with randomised controlled trials and meta-analyses of randomised trials ranking at the highest levels. In contrast, cognition-based medicine (CBM) is based on the physician's recognition of causality of a treatment in the individual patient. A careful examination of potential sources of misinterpretation reveals that none of these approaches enforces fairness of comparisons: Randomised controlled trials often cannot be generalised, meta analyses are based on retrospectively-selected studies and exposed to the ecological fallacy, and CBM approaches are not sufficiently controlled. Thus, fairness does not result from the choice of a perfect method or from the tailoring of the evaluation method to the therapy under examination. Rather, a complementary approach of different evaluation methods with all treatments to be compared should be used, with open discussion of strengths and weaknesses of the approaches applied in each particular case. PMID- 16044785 TI - [Therapy research in times of patient preference]. AB - The Declaration of Helsinki bears the consequence that in the future reference controlled clinical trials will increasingly replace placebo-controlled trials. This has enormous implications on research in complementary and alternative medicine: strong preferences for or against the compared therapies will prevent patients to consent to randomisation. In these circumstances a number of alternative designs have been proposed, based on the combination of preference and randomisation, but some of them suffer from severe methodological problems. Others are internally valid but unfeasible in practice and difficult to interpret. Thus, in case of strong patient preferences, we suggest to conduct non randomised trials but to give special interest to advanced statistical methods (e.g. structural equation models) which must replace simple two-group comparisons. PMID- 16044786 TI - [Which evaluations are fundamental for medical training in pharmacotherapy?]. AB - In Germany the medical profession is regulated by the state. The state defines the contents of the medical training within the clear boundaries of scientific medicine, which differs from science or biological medicine. The concepts and practice of many complementary therapeutic or diagnostic methods such as homeopathy do not fit within this state-set frame. Medical students, therefore, must be well acquainted with accepted scientific criteria by which we evaluate the value and validity of various therapeutic approaches. Lecturing pharmacotherapy implies posing questions such as: How do I come to statements which can be falsified? Within which constructs are they valid? Within which social context are they meaningful? Which aspects of reality must stay out of focus in order to reach such statements? This will help students to understand the frequent cognitive dissociations between doctors and patients as to concepts of disease and therapy, and the potential reasons why patients often prefer alternative medicine. Alternative methods can be openly discussed and evaluated on the basis of a critical methodology. For the sake of optimal patient care, "pluralistic" approaches towards setting valid therapeutic rules and standards are no alternative during the periods of basic medical training. PMID- 16044787 TI - [Is there a rationale for pluralistic evaluation models? Limitations of randomized clinical trials]. AB - Randomized clinical trials (RCTs) are believed to deliver the highest level of clinical evidence, and other types of evidence are rarely integrated in therapy assessments. Under ideal circumstances, RCTs are perfect tools of therapy evaluation; however, in clinical reality they suffer from a variety of limitations. Their prioritization can distort public health regulation because of commercial bias, career bias, bias of large numbers, mediocrity bias, and bias of therapy preferences; also, they can lead to discrepancies between clinical research and routine medical practice, to ethical problems, and to divergent and asymmetrical results. Moreover, the seemingly simple therapy assessment based on RCT results bears the danger of their endorsement and misuse by non-medical personnel, especially bureaucrats. To meet the demands of the complex reality of medical care, the physician's judgment should be methodologically professionalized and re-integrated into the process of generating medical knowledge; fixed hierarchies of medical evidence should be de-emphasized in favor of information syntheses from different types of (quality assessed) evidence. PMID- 16044788 TI - [Cognition-based medicine from the viewpoint of evidence-based medicine]. AB - The anthroposophic medicine plays a prominent role among the special forms of therapy. The concept of a cognition-based medicine (CBM)--in critical analogy to the evidence-based medicine (EBM)--has emerged from the circle of promoters of this type of therapy. The present paper attempts to identify common and different features of EBM and CBM. Criticisms of CBM to EBM are presented and addressed. The central aspect of the CBM concept is the single-case recognition, in analogy to the early gestalt-psychological considerations of Dunckers in the nineteen thirties. The EBM can certainly accept the critical relevance and justification of this concept, particularly with regard to the constantly educational (self)evaluation of the clinical decision. Individual causal recognition, on the other hand, is unable to replace the summatory evaluation of controlled studies. In certain single cases, the CBM makes "evident" the effect of a given therapy, on the other hand it is unable to establish a generally valid efficacy, which is so critical for the orientation of patients, physicians, and the socially-based healthcare system. PMID- 16044789 TI - [What is cognition-based medicine?]. AB - Cognition-based medicine (CBM) is a newly-developed methodological system of scientific medicine. Its primary element is the criteria-based assessment of therapeutic causality at the level of the individual patient. Principles and criteria of single-case causality assessment have been analyzed and explicated. CBM enables a methodological professionalization of clinical judgment, as well as the explication of physician experience and expertise. CBM study designs expand the current range of clinical research, extending from criteria-based causality assessment in single cases to new forms of cohort evaluations. PMID- 16044790 TI - [Opportunities of conventional evaluation in complementary and alternative medicine]. AB - Patients frequently require and utilise complementary and alternative medicine in spite of the present lack of evidence of effectiveness for many methods. However, convincing research based on conventional study types (cross-sectional studies, observational studies, comparative studies, randomised controlled trials, and meta-analyses) allows a scientifically sound assessment, e.g. for homeopathy, acupuncture, and anthroposophic medicine. The established study methodology is, therefore, applicable to complementary and alternative medicine; the available results should be the basis for recommendation of effective clinical strategies as well as for health policy decisions. PMID- 16044791 TI - [The physician between guidelines and patient individuality]. AB - Guidelines and individual patient care are not contradictory but rather complementary. They provide orientation to physicians in situations of complexity and lack of clarity. In addition, they simplify and speed up the process of practical implementation of new medical knowledge in the patient care. Guidelines are also a systematic principle that helps to recognize and avoid diagnostic and therapeutic uncertainty, as well as to protect patients. Furthermore, well understood guidelines mean that the physician is less burdened and can dedicate himself to something currently missing in the physician-patient relationship: time for comprehensive medical interviews. PMID- 16044792 TI - [The viewpoint of complementary medicine]. AB - Patients have more therapeutics choices than ever before--at least theoretically- and they demand the choice. On the othe hand, physicians are increasingly obliged to treat their patients according to guidelines. In these guidelines, complementary medicine is usually underrepresented, due to the frequent argument that adequate studies are supposedly lacking. Indeed, compared to the enormous number of studies with chemically-defined drugs, the investigational activity in complementary medicine is rather limited. This is based o methodological and especially financial problems, but excellent studies are nonetheless available. The rules of free-market economy are being increasingly applied in medicine. This may result in the fact that patients change to a status of consumers who are no longer aware of their true necessities, with physicians changing into a status of mere wish fulfillers. However, recent studies show that the patients already react against this trend by either emphasising their autonomy and self responsibility, or by switching to therapeutic strategies (including complementary therapies) independent of their physicians. In both cases, the use of physician-based complementary medicine can be a supportive and stabilising measure in the treatment process. Therefore, these measures should be mentioned more frequently in medical guidelines. PMID- 16044793 TI - [Patient's individuality and application of guidelines in surgery]. AB - Individual treatment decisions can become considerably conflictual in view of the co-existence of medical professional guidelines, recommendations based on evidence-based medicine (EBM), and juridical and economical directions. Medical guidelines are not subject to an external review process; also, due to reduced practicability, the surgeons' compliance with guidelines remains relatively low. Surgical treatment strategies can rely on randomized clinical trials (RCTs) in approximately 20% of the surgical procedures and on non-randomized trials in approximately 70% of the cases. No evidence is given in approximately 10% of the cases. Specific problems of implementation of EBM in surgical disciplines are represented by the difficulty of standardized procedures, the heterogeneity of the population, the impossibility to conduct double-blinded RCTs, a low statistical power, and a publication bias. Since individual diseases cannot be reduced to surgical cases manageable only by the application of guidelines, adequate treatment of individual patients requires the critical application of both external evidence and surgeon expertise (internal evidence). PMID- 16044794 TI - [University medicine, peripheral medicine, care research]. PMID- 16044795 TI - [Chronic cough: a practical approach]. AB - Chronic cough (more than 8 weeks) is a frequent symptom (30 millions consultations/ year). The most encountered causes are: asthma, gastro-oesophageal reflux, post nasal drip. Practically we propose the following approach: 1. clinical history, physical examination, chest-X ray, spirometry; 2. to exclude a post infection cough or secondary to an ACEI; 3. in case of high clinical probability of asthma, post-nasal drip, gastro-oesophageal reflux, to treat adequately. In case of negative clinical probability or unsuccessful treatment, metacholine test, oesophageal studies, PEF recording, CT thorax, bronchoscopy, CT sinuses are the most useful tests, using clinical history as guide. Using such an approach, treatment is successful in the vast majority of cases. PMID- 16044796 TI - [Drug-induced lung disease in general practice]. AB - Adverse medical side effects are an important health issue causing morbidity and mortality. Lungs, being a complex system are also involved. Cough, bronchospasms and buccopharyngeal oedema are the main drug-induced reactions at the airways level. With regard to pulmonary parenchyma, this can result in almost all forms of interstitial lung disease such as usual interstitial pneumonia, non specific interstitial pneumonia, lung oedema, eosinophilic lung disease. Certain drugs are often involved and cause specific clinical pictures (amiodarone, methotrexate). Lung vasculature, pleura and thoracic wall can also be affected. Drug-induced pulmonary disease is an exclusion diagnosis and the main therapeutic action is to stop administration of the suspected drug. On the Internet www.pneumotox.com is a helpful tool for the clinical practice. PMID- 16044797 TI - [Obstructive sleep apnoea syndrome]. AB - Sleep apnoea syndrome is a frequent disease, occurring in men aged more than 30 years and in postmenopaused women. Its prevalence is more than 1% in men and between 0,3%-0,5% in women. It is an important public health problem. Diagnosis is made by polysomnography, which is considered as a gold standard, or by polygraphy. Sleep apnoea syndrome must be looked for by clinicians in presence of symptoms such as sleepiness, snoring, or behavioural problems, because these patients have a high risk not only of driving crashes, but also of arterial hypertension, brain strokes, and myocardial infarction, without treatment. PMID- 16044798 TI - [OSAS and driving capacity]. AB - Obstructive sleep apnea syndrome (OSAS) is a very frequent clinical problem, sleepiness being one important symptom. Car driving necessitates attention and concentration. That why OSAS patients are at high risk of having car crash. But more than 60% of these patients will never have a car crash. CPAP treatment is very efficient also to correct this risk. Despite utilisation of driving simulator, we are actually unable to detect, in the OSAS population, the individual who will have a crash. To diagnose OSAS very early and to treat efficiently remain the key of security. PMID- 16044799 TI - [Medical help for smorking cessation: the pneumologist's point of view]. AB - Fighting against tobacco requires two main strategies: first, political action aims at preventing smoking beginning and to change the falsely promoted positive social image of smoking behaviour; second, the individual action of each practitioner is indeed very efficient to increase the cohort of ex-smokers. We have at our disposal, powerful, well codified and effective therapies to help patients to quit smoking. Their use ought to be focused on the following aims: detection of smokers, evaluation of motivation to quit and dependency, prescription of nicotine substitutes or bupropion, psychological support, patient follow-up and prevention of smoking relapse. In patients with COPD, smoking cessation is the only proven and effective intervention to stop disease progression and decrease COPD mortality. PMID- 16044800 TI - [Platypnoea and orthodoexia syndrome, a rare cause of dyspoea]. AB - Platypnoea-orthodeoxia is a rare syndrome of postural hypoxaemia accompanied by breathlessness. The dyspnoea is induced by upright posture. The definitive diagnosis is usually made by contrast echocardiography. The treatment of choice is surgical closure of the intracardiac communication which may result in dramatic symptomatic improvement. PMID- 16044801 TI - [Empathy vs antipathy or the Godel theory]. PMID- 16044802 TI - [Null can be expected to be ignored . . . why the need clause has been renewed for three years!]. PMID- 16044803 TI - [A fifth human retrovirus discovered at the Pasteur Institute of Paris]. PMID- 16044804 TI - [The pub, the PS and the doctor]. PMID- 16044805 TI - [The lay republican and therapeutic cloning]. PMID- 16044806 TI - [Avian flu pandemic: act before it's too late]. PMID- 16044807 TI - [The brain and liberty]. PMID- 16044808 TI - A study on trend of relapse in leprosy and factors influencing relapse. AB - A retrospective analysis of data pertaining to the rural field operation area of the Central Leprosy Teaching and Research Institute, Chengalpattu, Tamil Nadu, was carried out to determine the magnitude of relapse after MDT and its significance with other variables. The study included 3248 leprosy patients who have successfully completed treatment during 1987-2003, of whom 2892 were PB and 356 MB cases. A total of 58 cases of relapse was reported which gives a crude cumulative relapse rate of 1.78% for the 16-year period of follow-up and the rates for PB and MB were 1.9% and 0.84% respectively. With respect to PB cases, 68% of relapses were reported in the first 3 years of RFT. The person-year relapse rate was highly significant with regard to the number of skin lesions (p<0.0002) and nerve involvement (p<0.0002). The person-year relapse rate did not differ significantly between PB and MB leprosy, male and female, and child and adult cases. RFT year cohort relapse rate reveals that the introduction of MB-MDT regimen for PB leprosy had resulted in the reduction of relapses among PB cases after 1998. The relapse rate with reference to the time gap after RFT reveals that relapse declines with passage of time after RFT. The risk of relapse was very low in both PB and MB leprosy which fact emphasizes that proper counselling about signs and symptoms of relapse during RFT is adequate to combat the problem. A majority of relapses occurred in the first three years after RFT. The number of skin lesions and involvement of nerves were the main risk factors for relapse. PMID- 16044809 TI - Use of reverse transcription polymerase chain reaction for the detection of Mycobacterium leprae in the slit-skin smears of leprosy patients. AB - The relevance of bacterial index (BI) for understanding the prognosis of leprosy patients on treatment has been extensively debated, as it does not give a very clear idea of the viability of the bacteria in patients under treatment. Here we used slit-skin smear samples of leprosy patients to test the suitability for studying viability of Mycobacterium leprae using reverse transcription polymerase chain reaction (RT-PCR). For this purpose, we recruited 13 multibacillary (MB) leprosy patients (8 lepromatous and 5 borderline lepromatous). Of these, 7 were relapse cases, 3 were under treatment (MB-MDT), 2 were new cases and 1 had completed treatment. We carried out extraction of RNA using Trizol reagent (Life Technologies, UK) from the slit-skin smear samples from these patients. The RNA preparation was then used for the RT-PCR using Mycobacterium leprae-specific primers for the fragment of 16s ribosomal RNA gene. Samples from both the new cases, 4 suspected relapse cases and 1 patient under treatment showed positive RT PCR results. Other 6 patients whose smear samples did not show any amplification by RT-PCR were on MB-MDT from 8 to 30 months. The usefulness of the technique needs to be validated using mouse footpad technique and also should be more extensively explored for studying the viability of M. leprae, the efficacy of treatment and the presence of other mycobacteria in the slit-skin smear samples. PMID- 16044810 TI - Leprosy scenario in southern part of Dakshina Kannada district, Karnataka, after 16 years of control work. AB - This article examines the changes that occurred in epidemiological indices over a period of 16 years following the introduction of MDT in Ullal town, south of Mangalore city, having a population of 130,000. The analysis indicates that new case-detection rates and prevalence rates showed a declining trend due to shorter duration of treatment with MDT. There was a ten-fold reduction in the prevalence rate during the first 6 years, from 23 in 1987 (230 cases) to 2.76 per 10,000 (29 cases) at the end of 16 years. The number of nmultibacillary cases among the newly detected cases showed a downtrend (from 28 cases in 1987 to 5 in 2001). The number of newly detected cases presenting with single lesion also showed a declining trend. PMID- 16044811 TI - Fnac study of histoid nodule: an early tool for diagnosis. AB - Histoid lesion, a variety of lepromatous leprosy, is due to alteration in the growth pattern of Mycobacterium leprae, possibly due to loss of immunity in localized areas. The distinction is based on cellular morphology by demonstrating pallisading arrangement of multi-layered spindle-shaped histocytes. Cytodiagnosis by fine needle aspiration cytology is therefore an early tool to recognize the histoid variety, differentiating it from a conventional LL module, as it is a simple and less traumatic procedure. PMID- 16044812 TI - Bilateral seventh nerve palsy--a manifestation of polyneuritic leprosy. PMID- 16044813 TI - Co-existence of lepromatous leprosy and myasthenia gravis. AB - Various circulating autoantibodies are known to occur commonly in patients with lepromatous leprosy. However, the association with autoimmune diseases has rarely been reported. One such association of lepromatous leprosy with myasthenia gravis is being reported. PMID- 16044814 TI - Ganglion cyst of the proximal tibiofibular joint causing foot drop-diagnostic problem of differentiating from neuritic leprosy. AB - A six-year old boy presented with pain around the knee joint and abnormal gait of one month duration. There was no history of hypopigmented anaesthetic patches, neuritis or family history of leprosy. Clinical examination revealed a localized cystic swelling of 1 x 1 cm in size in the region of left common peroneal nerve, with sensory loss on the lateral aspect of the left leg which was mistaken for a nerve abscess. PMID- 16044815 TI - Molluscum contagiosum-like lesions in lepromatous leprosy. AB - A 32-year-old male patient presented with multiple, asymptomatic, papulonodular lesions some of which were umbilicated simulating the clinical features of molluscum contagiosum. On examination the patient was found to be suffering from lepromatous leprosy. Slit-skin smear showed a BI of 6+ and histopathological examination of the nodular lesion showed features of lepromatous leprosy. The patient was treated with MDT-MB and improved remarkably in a few months. PMID- 16044816 TI - Guillain-Barre syndrome 1N leprosy patients. AB - Leprosy commonly affects the peripheral and cranial nerves. This involvement often manifests with peripheral neuropathy, occurring as a result of direct invasion of the nerves by lepra bacilli. Immune-mediated neuropathy occurring in leprosy, as part of lepra reaction, has been described earlier. We report here two cases of Guillain-Barre syndrome occurring in patients with leprosy who did not have any obvious evidence of lepra reaction. PMID- 16044817 TI - Necrotic erythema nodosum leprosum. AB - A 30 year-old farmer presented himself with mild degree fever, joint pain and multiple ulcers of 15 days' duration. Ulcers were multiple, well-defined, irregular in shape with necrotic base and elevated, sloppy margins over anus, forearms, wrists, buttocks, lower half of thighs and shins. A few of the ulcers were covered with black-coloured scab. They were tender and with serosanguinous discharge. Black atrophic scars and ill-defined, irregular, brown black-pigmented macules of various sizes were present over thighs and legs. Excision biopsy of the ulcer showed features of erythema nodosum leprosum. The patient was treated with systemic corticosteroids. PMID- 16044818 TI - Borderline tuberculoid leprosy localised over a scar. PMID- 16044819 TI - Cranial nerve involvement in leprosy. PMID- 16044820 TI - What is lazarine leprosy? Is it a separate entity? PMID- 16044821 TI - Lazarine leprosy--revisited. PMID- 16044822 TI - Regional conference on leprosy, Chennai 15-16 May, 2005. PMID- 16044823 TI - From maternal mortality to malignancies . . . what lies in between? PMID- 16044824 TI - Improving knowledge of antenatal care (ANC) among pregnant women: a field trial in central Java, Indonesia. AB - The need for reducing maternal mortality has become a paramount concern in developing countries including Indonesia. One of the strategies for reducing maternal mortality in Indonesia is the provision of antenatal care (ANC). Previous studies have reported the advantages and disadvantages of ANC. The purpose of this study is to ascertain if a new approach to ANC can improve pregnant women's knowledge of its benefits. An experimental design with 60 pregnant women from 10 cluster villages is used in this study. The intervention group received the new approach to ANC, while the control group received routine ANC. The findings show that the improvement of knowledge in the intervention group is significant particularly in the knowledge about healthy pregnancy (p=0.012), pregnancy complications (p=0.01), safe birth (p=0.01) and taking care of the newborn (p=0.012). The improvement of knowledge was significantly influenced by the respondents' educational back ground (p=0.002) and socio economic status (p=0.027). This study recommends that the new approach to ANC be considered to educate pregnant women regarding safe birth and it is considered as one of the strategies that may be adopted to reduce maternal mortality. PMID- 16044825 TI - Cardiovascular risk factors in Iranian adults according to educational levels: Isfahan healthy heart program. AB - There is abundant evidence that the socioeconomic status (SES) is inversely associated with CVD risk factors. The objective of this study is to describe the distribution of CVD risk factors according to educational level in Iranian adults. A cross-sectional survey was performed on subjects over the age of 19 in three cities. Demographic data, as well as information on educational level and smoking habits was obtained and then height, weight and blood pressure measurement and blood sampling were arranged. Data was analyzed by the Mantel Haenszel, Kendalls T correlation and multivariate analysis test. Of 9587 subjects, 48% were men and 52% were women, with mean ages of 39.0+/-15.3 and 38.8+/-14.5 years, respectively. All CVD risk factors showed an inverse relationship with educational level in all subjects, except for smoking and low HDL-C in women. High TC and LDL-C and low HDL-C were inversely related to educational level (P<0.05); however, this relationship was not significant with Mantel-Haenszel test in men. The association between CVD risk factors and educational level in women was stronger than in men. The differences found between CVD risk factors with educational level are important and should be considered in programs designed to increase level of education in order to lower CVD risk factors. PMID- 16044826 TI - Self-disclosure of HIV status: perception of malaysian HIV-positive subjects towards attitude of dental personnel in providing oral care. AB - The objective of this study was to assess the perceptions of Malaysian HIV positive subjects towards the attitude of dental personnel in providing oral care to them. The study design was cross-sectional with the sampling frame comprising of 27 Government Drug Rehabilitation Centres throughout Malaysia. A convenience sample was then taken from 20 centres with the highest enrolment of HIV-positive subjects. A self-administered questionnaire was used to elicit information on the perception of HIV-positive subjects towards the attitude of dental personnel in providing oral care to the patient with HIV-positive. The study sample consisted of 509 HIV-positive individuals with a mean age of 31.3+/-12.9 years old. Of these, only 15.1% attended a dental clinic after confirmation of HIV-positive status. The study demonstrated that 67.5% of the HIV-positive subjects disclosed their status voluntarily to the dentists and majority of the dentists (76.9%) did not show any negative reaction on knowing their HIV positive status. There was also no difference in the attitude of auxiliary staff toward the above disclosure. In conclusion, the study showed that oral health care personnel are more receptive to the HIV-positive subjects receiving dental care and treatment. PMID- 16044827 TI - Iron deficiency anaemia and child development. AB - A developmental test was carried out to evaluate the effect of iron deficiency anaemia on child development by using Bailey Scale of Infant Development (BSID) and Enjoji Scale of Infant Analytical development (ESID). The subjects were 54 children, divided into three groups. Group 1 consisted of 15 infants with anaemia; Group 2: 10 infants with a past history of anaemia and Group 3: 29 healthy normal infants without anaemia formed the control group. The characteristics of infants and their family background were not different among the three groups except for the male/female ratio. The mean mental developmental index (MDI) and psychomotor developmental index (PDI) of Group 1 and Group 2 were lower than that of control group using the BSID test. The mean speech development quotients of Group 1 and Group 2 were lower than that of control group in ESID. These tendencies were observed after subjects were stratified by sex. Therefore, the results of the study suggests that iron deficiency anaemia may affect child development especially speech development. PMID- 16044828 TI - Incidence of colon cancer in Tianjin, China, 1981-2000. AB - The objective of this study is to describe and analyze the incidence trend of colon cancer in Tianjin, China from 1981 to 2000. Tumour cases were coded by ICD 9 in this study. Incidence rates were calculated by five-year age-groups as well as sex and year of diagnosis. From 1981 to 2000, the total number of colon cancer cases ascertained in urban Tianjin was 4954, including 2547 males and 2407 females. 67.88% colon cancer cases occurred in the age group 55-79 and age specific incidence rate reached its peak in the age group 75-79. The mean incidence rate of colon cancer during the 20 years was 7.01/100000 and this rate had been increasing constantly from 1981 to 2000. The average age at diagnosis was 62.41 years. An ascending trend was observed in the mean age at diagnosis of colon cancer from 1981 through 2000. As for the sex ratio, there was no clear trend exhibited. The incidence trend of colon cancer during 1981 to 2000 in Tianjin warranted a further research on its risk factors and prevention warranted. PMID- 16044829 TI - Infrared thermography to mass-screen suspected SARS patients with fever. AB - Fever greater than 38 degrees C is a cardinal sign of patients with the severe acute respiratory syndromes (SARS). To reduce the risk of nosocomial cross infections, screening all patients and visitors who visit hospitals and clinics for fever at the entrance of every hospital building has become a standard protocol in Taiwan during the SARS epidemic from mid-April to mid-June 2003. We used a digital infrared thermal imaging (DITI) system (Telesis Spectrum 9000 MB) to conduct mass screening of patients and visitors who entered the hospital to identify those with fever. The DITI system has two components: a sensor head and a PC imaging workstation. The sensor head is an optic-mechanical device which consists of imagining optics for focusing the infrared source information on the infrared detector. The infrared images are further converted into electrical signals, which are then processed for real-time display on the monitor. During the period from April 13 to May 12 2003, 72,327 outpatients and visitors entered Taipei Medical University-Wan Fang Hospital, Taipei, Taiwan. A total of 305 febrile patients (0.42%) was detected by infrared thermography. Among them, three probable SARS patients were identified after thorough studies including contact history, laboratory tests and radiology examinations. The findings suggests that infrared thermography was an effective and reliable tool ideal for mass-screening patients with fever in the initial phase of screening for SARS patients at a busy hospital which sees approximately 3,000 outpatients every weekday during the SARS epidemic. PMID- 16044830 TI - A quantitative food frequency questionnaire for women in southeast China: development and reproducibility. AB - This study describes the development and reproducibility of a 128-item quantitative food frequency questionnaire (FFQ) to measure usual food consumption for women in southeast China. The FFQ was pre-tested using 51 Chinese women who recently migrated to Australia. Cronbach's alpha coefficient was 0.81 for internal consistency. The reliability of the FFQ was then assessed by another test-retest study. A sample of 41 women residing in southeast China was interviewed twice within 12 weeks. Intraclass correlation coefficients were moderate to high for mean food group consumption (0.43-0.96) and mean daily nutrient intakes (0.47-0.89). Kappa statistics for eating habits ranged from 0.27 to 0.89 in the test-retest. The mean ratio of energy intake to basal metabolic rate was 1.73 (S.D. 0.39) in both test and retest samples. The study confirmed that the FFQ method using standard containers is appropriate to assess dietary intake for women in southeast China. PMID- 16044831 TI - Wheat thresher agricultural injuries: a by-product of mechanised farming. AB - Farm mechanization has resulted in extensive use of wheat threshers on Indian farms. It has also increased agricultural injuries. A prospective study was undertaken for analysis of wheat thresher agricultural injuries and their remedial measures. Fifty two patients presenting with thresher injuries during the wheat harvesting season of March to June, 2003 were studied. A study-specific 14-point proforma was prepared to gather all possible information from site of injury to hospital records. Injuries were mostly of the upper limb and amputations accounted for most of these. Poor light arrangements, unskilled workers, drug / alcohol abuse, fatigue, poor designing and lack of orientation to work on these machines were the contributory factors to such injuries. The analysis emphasizes that the need of the hour is to decrease wheat thresher injuries through specific preventive approaches like improved designing, education, legislation, compensation and surveillance programmes. PMID- 16044832 TI - Smoking in adults in Sri Lanka: prevalence and attitudes. AB - A questionnaire was administered to 1565 adults to identify the prevalence of smoking and to assess respondents' attitudes toward smoking. Of men 41% were yearly smokers, 27.8% were monthly smokers and 21% were daily smokers. The corresponding figures for women were 3.4%, 2% and 0.6% respectively. Higher prevalence rates were observed among less educated, middle-aged men who were from underprivileged families. About 23% of men and 0.9% of women were monthly alcohol users. Alcohol use seems to be positively associated with smoking. A considerable portion of both smokers and non-smokers expressed a favorable attitude towards smoking habits of adult males (40.1% of smokers and 12.8% of non-smokers) and towards the tobacco industry (25.1% of smokers and 13.1 of non-smokers). Since the majority of people in Sri Lanka are non-smokers, attitudinal support given by them in promoting smoking in the society needs to be addressed effectively in smoking control interventions. PMID- 16044833 TI - The association between subjective and clinical indicators of health in prospective Vietnamese migrants. AB - We conducted a cross-sectional survey of 1669 prospective Vietnamese migrants who had applied to migrate to Australia to describe the association between self reported health status and several commonly used clinical indicators of health among prospective Vietnamese migrants. Participants were recruited from the International Organization for Migration's standardised medical screening program.' We found that clinical indicators of health are related to self reported health status among prospective Vietnamese migrants. Self-reported health status, which was assessed using a modified version of the Short Form-36 health survey, was significantly associated with clinical indicators of health, including the number of body system abnormalities identified during medical screening, evidence of tuberculosis on chest radiograph, and self-reported weight loss over the previous six months. These findings support the validity of self reported health status assessment among prospective migrants, although the assessment of subjective indicators of health during compulsory medical screening may be limited by reporting bias. PMID- 16044834 TI - [Biosafety. Natural contaminants of food mycotoxin]. AB - The present paper review the current knowledge on principal mycotoxins of confirmed or potential relevance in human health with respect to occurrence, toxicology and risk assessment. PMID- 16044835 TI - [Assessment of diet of students from Orenburg region]. AB - The article this research is devoted to the Orenburg student youth diet. The revealed deficiency of major nutrients (protein, fats and especially carbohydrates), as vitamins (C, PP, E, D, B groups) and analysis of diet ration concerning basis nourishing components, vitamins, macro- micro elements (Zn, Cu, I, Cr, Se). PMID- 16044836 TI - [Estimation of nutrition of elderly and senile patients with surgical diseases at home]. AB - Analysis of estimation of nutrition 500 of elderly and senile patients with surgical disease at home. The diets of patients are characterised by foods rich of animal protein and carbohydrate. But a dificienny of meat, fish, dairy products frent, berries is marhed. The diets thas dificiency in rations of patients are characterized by irrational set basic food products, cannot provide daily needs for nutrients: animal protein, vitamins C, A, vitamins B group, beta carotene and mineral element--Ca, Mg, P, Zn, Se. Entering of nutrional substance, such as vitamins, and mineral elements with a food is reduced in dietary restrictions, deficiency means, residence in country-side. PMID- 16044837 TI - [The influence of dietary inulin and oligofructose on the cell-mediated and humoral immunity in rats]. AB - This paper reviews evidence for the immune-enhancing effects of prebiotics. The influence of inulin and oligofructose on the immune functions and the structure of immune peripheral organs in male Wistar rats was investigated after 4 months feeding. Rats fed isocaloric purified diets contained 10% inulin or oligofructose. The rats of control group had no feeding any prebiotics. The enlargement of a number of T-cells and increasing of major histocompatibility complex (MHC-II) molecule on the surface of an antigen-presenting cells of spleen, mesenteric lymph nodes, thymus was noted. The increasing of content interleukin-2 and interleukin-4 in blood was noted in rats fed inulin and oligofructose. The increased activity of peritoneal macrophages was confirmed by enhance superoxid anion production and phagocytosis. The dietary prebiotics do not influence on the apoptosis of immunocytes. The immune-enhancing effects of prebiotics was confirmed by morphological investigation of peripheral lymphoid organs. PMID- 16044838 TI - [Influence of chitosan feeding of laying hens on egg vitamin and cholesterol content]. AB - Chitosan feeding (10 and 20 mg per 1 kg body mass) of 19 week-age laying hens during 1.5 months caused a decrease in whole egg content of vitamin A for 13% and 20% (p < or = 0.05), vitamin E--for 30%, lutein--17% and didn't effect on vitamin B2 level. Chitosan intake lead to cholesterol content 1.5-2 fold decrease and didn't influence on egg yolk lipids concentration. Low dose chitosan-receiving hens had eggs with 1.8-fold increased egg yolk phospholipids level. The most optimal dose of chitosan for the improvement of eggs nutritive value was 10 mg. Under minimal loss in vitamins its administration lead to the pronounced cholesterol decrease and marked phospholipids elevation. PMID- 16044839 TI - [Application of nutritional formulare in the dietotherapy in patients with cardiovascular diseases]. AB - It was found that the including of nutritional formulae in the dietary therapy for patients with cardiovascular diseases and obesity has been evaluated, had a positive effect on the clinical presentation of disease, lipid metabolism and the dynamic of body composition of patients. PMID- 16044840 TI - [The treatment of coronary heart disease by beta-adrenoblockers or tiazide diuretics preparation in combination with vegetarian diet]. AB - Work make on 84 patients with coronare heart diseases were divided into two groups, equal quantity. The first groups were given athenolol (50 mg daily), the second--hypotiazide (25 mg daily). In every groupspart patients received an antiatherogenic lactoovovegetetarian diet, part--an standard antiatherogenic diet 10c. Time observation--24 daily. By the end of the treatment period with athenolol in backoground the vegetarian diet the level of total cholesterol decreased by 16%, low-density lipoproteins cholesterol decreased by 18%. In groups patients received an standard antiatherogenic diet these parameters practically did'nt change. In the vegetarian group the atherogenic coefficient (KA) decreased by 31%., while in the patients on standard antiatherogenic diet KA showed only a tendency for decreasing. By the end to the treatment period with hypotiazide the slight decrease in total cholesterol, KA levels and a slight increase in HDL cholesterol were observed only the vegetarian group. PMID- 16044841 TI - [Tea and coffee consumption among Russian population]. AB - Aim of present study was the getting data on black and green tea and coffee consumption among Russian population in 1995-2003. The consumption of beverages tea and coffee was estimated by 24-hour recall of dietary intake in course of Russian Longitudinal monitoring surveys of economics and health. About 82% of population have consumed the black tea and only 2.2%--the green tea. Mean daily black and green tea intake among all population were 446 ml and 10 ml, correspondingly. The prevalence of black tea consumption was decreased and green tea and coffee consumption were increased from 1995 to 2003. PMID- 16044842 TI - [Application of infusoria Tetrahymena pyriformis culture in quality method of estimation with a help of frozen raw material from Pacific herring fatty]. AB - Influence of refrigerated storage on quality of fatty raw material by the example of Pacific herring has been investigated. After freezing and during storage at different temperatures in samples of frozen Pacific herring wedetermined the microbiological parameters, the content of toxic compounds, changes of proteins and accumulation of products of lipids oxidation. In parallel it was determined the quality of frozen herring by method of biotesting using infusorian Tetrachymena pyriformis. It has been established, that during storage of frozen Pacific herring the loss quality of fish raw material takes plase. Results of biotesting completely correlate with the data of chemical analysis under the estimation of fish raw material quality of refrigerated storage. PMID- 16044843 TI - [Stability of ascorbic acid at it contract alloy in presence of fluoride ions]. AB - Aluminum utensils are considered as potentiAl source of dietary Aluminums. Report suggests that acidic food cooked or stored in presence of Aluminum foil contain high concentrations of Aluminum. Study on fluoride induced leaching of Aluminum from different pH. Higher concentrations of fluoride and lower pH enhance Aluminum leaching to a great extent. Evidence was obtained showing that after a 2 dyas exposure at room temperature in presence of floride NaF, Aluminum foil liberated nearly 1 mg/l of Aluminum, compared with less than 0.04 mg/l in absence of fluoride. There is reason to believe that in experiments with ascorbic acid NaF prevents the oxidation of ascorbic acid. PMID- 16044844 TI - [The use of natural hitamine-based natural substance at chronic cholecystitis]. AB - The results of application of hitamine, enterosorbent of marine origin, by the patients with chronic noncalculous cholecystitis in the remission phase are cited. It has been established that dietotheraphy in complex with hitamine renders the antiinflammatory and lipidcorrected action, favours the increase of remission terms. PMID- 16044845 TI - Antitrust risk: FTC/DOJ attack on the messenger model. PMID- 16044846 TI - Dental Day at the Capitol. PMID- 16044847 TI - A world of work to do. PMID- 16044848 TI - How stupid do they think we are? PMID- 16044849 TI - On the path before us. PMID- 16044850 TI - The endless learning curve: 2005 Table Clinic winners. PMID- 16044851 TI - RM matters: treating patients on anticoagulants. PMID- 16044852 TI - What's a dentist to do? Values, part four. PMID- 16044853 TI - Location, location, location, or, where the heck am I anyway? PMID- 16044854 TI - Killed in action: microbiologists and clinicians as victims of their occupation. Part 1: typhus. PMID- 16044855 TI - Threat of infection: microbes of high pathogenic potential--strategies for detection, control and eradication. AB - Infectious diseases due to microbes of high pathogenic potential remain a constant and variable threat for human and animal health. The emergence of new diseases or the re-emergence of diseases that were previously under control complicates the situation to date. Infectious disease research, which has undergone a dramatic progress in understanding disease mechanisms such as host pathogen interactions, is now focusing increasingly on new strategies for prevention and therapy. Significant progress has been achieved in the development of delivery systems for protective heterologous protein antigens and in veterinary vaccinology. A landmark of infectious diseases research is the chemical synthesis of genomes, a major new field of research referred to as "synthetic biology", that to date has resulted in the chemical synthesis of the poliovirus and of phage phiX174 genomes and their expression as infectious viruses. On the molecular level the evolution of pathogens and mechanisms of genome flexibility, which account for several pathogenic properties of infectious agents, have received increased attention. Bacterial toxins are an additional threat to human health and their interference with host cells and cellular functions is receiving more attention. PMID- 16044856 TI - Analysis of the growth phase-associated transcriptome of Streptococcus pyogenes. AB - Streptococcus pyogenes (group A streptococci, GAS) is a human pathogen which probably varies its multiplication rate and thus, growth phases in association with the type of infection caused in its host. To create a basis for future determinations of such associations, the genome-wide growth phase-related GAS transcriptome was assessed in the present study. Therefore, the published serotype M1 S. pyogenes genome sequence as well as the partially sequenced serotype M18 and M49 GAS genomes were used to produce DNA microarrays that carried 2256 oligonucleotide probes matching 3662 open reading frames (ORFs). With these microarrays, the transcriptome of the serotype M49 GAS strain 591 grown to the exponential, transition, and early stationary growth phases was assessed in seven independent experiments. The gained data were compared to real time RT-PCR assays. Data analysis was refined by a novel approach, i.e. grouping of expressed genes to four classes according to relative transcript abundance and gene functions. At the different growth phases, 86.7%, 79.5% and 55.7% of the at least 1883 ORFs contained in the serotype M49 genome were expressed above the defined detection level. Contrary to the general trend, transcript amounts of genes in the functional groups of transport and membrane proteins as well as stress response factors peaked at the transition phase. The most prominent changes in the transcript abundances were predominantly observed for sugar compound transport and turnover-related ORFs. The majority of known virulence genes had their maximum expression during the transition phase, consistent with the proposed associated change in virulence behavior of the bacteria. With these results, it will now be feasible to assess the in situ growth phase of a given GAS strain during any type of infection by measuring the expression of selected marker genes. PMID- 16044857 TI - Toll-like receptors TLR4, TLR5 and TLR9 on gastric carcinoma cells: an implication for interaction with Helicobacter pylori. AB - In the human stomach Toll-like receptors (TLRs) expressed by the gastric epithelium interact with Helicobacter pylori and mediate production of proinflammatory cytokines and chemokines during H. pylori infection. This results in chronic active gastritis, the background from which gastric carcinoma arises via the epithelial precursor lesions, intestinal metaplasia and dysplasia. Therefore, the question is arising whether gastric carcinoma cells are also able to interact with H. pylori. In this study, TLR4, TLR5 and TLR9 expression was investigated on tumor cells of gastric carcinoma and on its precursor lesions, intestinal metaplasia and dysplasia, by immunohistochemistry. Gastric epithelium with intestinal metaplasia (n=10) and dysplasia (n=3) expressed TLR4 and TLR5. TLR4 was strongly expressed by tumor cells of 17 out of 22 and TLR5 by tumor cells of all 22 patients with gastric carcinoma. TLR9, however, was not detectable in intestinal metaplasia or dysplasia and only focally in 6 out of 22 gastric carcinomas. In contrast to H. pylori gastritis, epithelial TLR expression in intestinal metaplasia, dysplasia and gastric carcinoma was diffusely distributed without subcellular polarization as demonstrated by confocal microscopy. This is the first study describing TLR expression on tumor cells of gastric carcinoma and its precursor lesions. Expression of TLRs enables gastric carcinoma cells to interact with H. pylori. As H. pylori can induce gastric carcinoma-promoting factors, such as IL-8, via epithelial TLR expression, TLR expression by gastric carcinoma cells may have a dangerous potential. PMID- 16044858 TI - Molecular analysis of intrafamiliar transmission of Moraxella catarrhalis. AB - The possible intrafamiliar transmission of Moraxella catarrhalis was evaluated in 3 pairs between children and their parents, and 8 pairs between siblings from 11 families. Of the 22 isolates, all were found producing beta-lactamase. Molecular typing by pulsed-field gel electrophoresis (PFGE) with Not I and Spe I showed that the PFGE patterns in 2 of 3 pairs between children and their parents, and 4 of 8 pairs between siblings were indistinguishable and those of the remaining pairs were different. These data indicate a possible high rate of intrafamiliar transmission of M. catarrhalis. PMID- 16044859 TI - A question of faith. Interview by Nick Lipley. PMID- 16044860 TI - Tapping into talent. PMID- 16044861 TI - Close encounters. PMID- 16044862 TI - Developing the ability to lead. PMID- 16044863 TI - Managing change initiatives in clinical areas. PMID- 16044864 TI - Unintended consequences. PMID- 16044865 TI - [Cholangiocellular carcinoma--clinical features and surgical treatment]. AB - Cholangiocellular carcinoma is the second on rate primary liver cancer (7-10%), after hepatocellular carcinoma--(80-85%). The aim of this study is to present some diferrencies in clinical and pathological features in comparison to other liver tumors. MATERIALS AND METHODS: In the period 1991-2002 years, 105 patients with primary liver cancer were operated; 9 of them (8.5%) had cholangiocellular carcinoma (CCC). The men were 5, the women--4, and the mean age 51 years. Hepatic resection was performed in 5 patients, in 1--biliary drainage, in 3--explorative laparotomy. RESULTS: Until 30th postoperative day there was no death in resected patients, but one--the drained patient. Morbidity was: ascites, hepatocellular insuficiency, fever. One patient is alive more than 4 years. CONCLUSIONS: Cholangiocarcinoma affects young people, sometimes they are jaundiced; tumours become big, because asymptomatic grow. Hepatic resection remains to be the best therapeutical option in nonjaundiced patients. PMID- 16044866 TI - [The Studer orthotopic bladder after radical cystectomy--observation of more than 40 patients]. AB - PURPOSE: To perform our 5-years experience in 40 patienets with orthotopic bladder substitution in Studer's technic follow radical cystectomy. MATERIALS AND METHODS: We created orthotopic neobladder using segment of terminal ileum in Studer's fashion in 40 patients. To decrease peristaltic and respectively bowel pressure, isolated segment was opened longitudinal and crossed sutured. This way we created a low pressure urinary reservoir. RESULTS: All patients were completely continent. Bladder capacity was 400-700 ml. At 4 years followup the patients didn't require correction in their therapy for metabolic disorder. Eight of the patients undergone this procedure died in 3-50 monts postoperatively for pelvic recurrence. CONCLUSION: Studer's orthotopic neobladder is an exellent alternative for patients undergone radical cystectomy. Thechnical ability decreases the operating time and surgical complications significantly. PMID- 16044867 TI - [Surgical treatment of hydrocele--modern aspects]. AB - Basically no one has ever introduced into practice any better procedure in cases of hydrocele than those of Winkelmann and Bergman. We recommend always a resection of the covering sheets of the testis as a profilactic measure against recurrency. Our study included 192 patients with hydrocele who were operated during the period 2000-2001 in the Clinic of Urology, Medical University of Sofia. The surgical procedures included: Winkelmann's procedure--in 118 cases (61,45%); Bergman's procedure--in 63 cases (32,81%);Orchiectomy--in 3 cases (1.56%) of total testicular atrophy; Combination of hydrocele and co-existing groin hernia was found in 4 cases (2,08%); Orchiectomy--in 1 case (0.52%) of bleeding which lead to hematoma on the 4th postoperative day; Reoperation--in 1 case (0.52%) of compression of the testicular blood vessels (after Winkelmann's procedure); Suppuration of the operative wound-- in 2 cases (1.02%) after Winkelmann's procedures. The authors consider that: 1. Winkelmann's and Bergman's procedures prooved to be quite satifactory as surgical treatment of patients with hydrocele. 2. Diagnosis and surgical treatment are easy to be performed. Postoperative complications are possible in cases of poor surgical technics, damaged local status and shortened time for hemostasis. 3. The well-known method of Lord was not used in our practice, but we do not deny it. PMID- 16044868 TI - [Liposuction in outpatient/one-day surgery]. AB - On the basis of 126 cases it has been demonstrated that liposuction is a surgical procedure, which can be performed safely on an outpatient basis only under precise conditions--patient class ASA I, recovery room, permanent phone connection between the doctor and the patient after leaving the facility. The suction of more than 2500-3000 ml fat tissue increases the hazards associated with this type of operation. The regional epidural anesthesia is perfectly suitable for the outpatient/one-day surgery. PMID- 16044869 TI - [Reconstruction of the large defects of the scalp with local fastiocutan flaps]. AB - No depending on the reason the large defects of the scalp, denuded from periost calvaria or without calvaria are difficult problem for reconstruction. There are several methods for reconstruction: local, distant pedicled and free flaps. Local fastiocutan flaps should be consider as a first step when planning scalp reconstruction. They are stable coverage and prepared by single technique with minimal functional loss and rapid patien recovery. In this paper we present 14 cases of this technique and discuss the advantage of the local fastiocutaneus scalp flaps. PMID- 16044870 TI - [The application of argon plasma coagulation in thoracic surgery: principles, surgical technique and clinical results]. AB - BACKGROUND: In recent years, together with the well-known high-frequency electro coagulation, the application of plasma coagulation has been also introduced in the clinical practice. The argon plasma coagulator (APC) is one of the representatives of this surgical technique. By its nature, the APC represents a non-contact electrothermal tissue coagulation, combining the principle of the augmented surface and enhanced autogenous haemostatic mechanisms. AIM: The main objective of this study was to evaluate whether APC is an effective and safe modality in the open pulmonary surgery. MATERIALS AND METHODS: For the period from 01.01.2003 to 30.01.2004 year, in the Clinic of Thoracic Surgery, we have applied the technique of APC to 15 patients. The distribution by sex was: 10 males and 5 females. According to the nosological units, the distribution was as follows: pulmonary carcinoma in 3, pulmonary echinococcosis in 4, pleural empyema in 6, pulmonary abscessus in 1 and esophageal ahalasia in 1 patient. In our practice, we have used an argon plasma coagulator of BERCHTOLD GmbH. A power setting of 20W with exposition time 15 s and an argon gas flow setting of 1,5-2 1/h have been used in our series. Energy dose applied in our patients didn't exceed 300 J/cm2. RESULTS: The results we have obtained demonstrate the following fundamental advantages of APC: a possibility to work with long electrode--tissue distance; a possibility for large surface coagulation as well as coagulation under variable angle, limited and well controlled depth of penetration, substantial reduction of carbonization; regular distribution of the energy over the whole coagulating surface, a possibility to treat effectively larger bleeding surfaces. CONCLUSIONS: Although initial, our experience gives us the confidence to recommend the use of APC as an effective and safe procedure in the pulmonary surgery. PMID- 16044871 TI - [Diagnosis of necrotizing fasciitis]. AB - In our surgical practice in the last few years, we notice an increase of severe soft tissue infections. One of the most damaging soft tissue infections we consider the necrotizing fasciitis. The mortality is high and the diagnosis is difficult. The deep necrotizing infection leads to vascular occlusion, ischemia and tissue necrosis. We notice a loss of sensibility of the affected areas because of the damaged nerve extensions. In the Clinic of Septic Surgery in Multipurpose Hospital for Active Treatment and Emergency Medicine "N. I. PIROGOV", Sofia, Bulgaria, on the base of 32 cases with necrotizing fasciitis we determined basic diagnostic signs. Those signs can be of assistance to surgeons in their medical practice who have or will experience this severe infection requiring urgent surgical management. PMID- 16044872 TI - [Antibacterial therapy of anaerobic surgical infections]. AB - The necrotizing soft tissue infection caused by anaerobic microorganisms or by mixed aerobic-anaerobic flora is one of the most severe infections in surgical practice with a mortality rate up to 75%. Of great importance for successful treatment are the early diagnosis followed by aggressive debridement and adequate antibacterial therapy. Depending on the studies of 358 isolates made in the Clinic of Septic Surgery, we specify the most often used antibiotics in cases of severe anaerobic surgical infection. It is established that the incidence of this severe infection is unpredictable and its spreading depends on the immune answer the concomitant illnesses and the previous undertaken surgeries of the patient. PMID- 16044873 TI - [Cytomorphological features of the fatty tissue tumors of the liver]. AB - The necrotizing soft tissue infection caused by anaerobic microorganisms or by mixed aerobic-anaerobic flora is one of the most severe infections in surgical practice with a mortality rate up to 75%. Of great importance for successful treatment are the early diagnosis followed by aggressive debridement and adequate antibacterial therapy. Depending on the studies of 358 isolates made in the Clinic of Septic Surgery, we specify the most often used antibiotics in cases of severe anaerobic surgical infection. It is established that the incidence of this severe infection is unpredictable and its spreading depends on the immune answer the concomitant illnesses and the previous undertaken surgeries of the patient. PMID- 16044874 TI - [Surgical methods for reconstruction of the temporomandibular joint and lower jaw in ankylosis and micrognathism]. AB - The treatment of the temporomandibular joint ankylosis is a challenge for the oral-facial surgery. In children the ankylosis is often followed by micrognathia and disturbances in the balance of the orofacial system and the whole organism and it makes the problem more serious. All the surgeons who have ever treated ankylosis, had theirs disappointments. The question about the surgical technique and when does it have to be made is still actually. Using the literature we try to represent the most useful techniques for reconstruction of the TMJ and the lower jaw in case of ankylosis and micrognathia We hope to optimize the treatment and to make the life of these patients better. PMID- 16044875 TI - [Health-related quality of live in breast cancer patients--actuality and problems]. AB - Women with early breast cancer have three surgical options fir this current moment: conservative treatment (lumpectomy, quadrantectomy); mastectomy alone and mastectomy plus reconstruction (immediate or delayed). When randomized trials showed oncological safety of breast--conservative treatment compared with mastectomy, the question of cosmetic results assessment become actual, because of its impact on quality of live. The current state of the art permits achievement of good oncological results and good cosmetics ones. There are some technical factors affecting cosmetic results after breast--conservative treatment: orientation of resection; the breast / resection ratio; tumor localization; orientation of axillary dissection. Combination of extended resection (quadrantectomy) plus axillary dissection through the same incision results in cosmetic failure. After operation there is a remarkable breast asymmetry, retraction of the mammilla and inferior contour distortion. The main reasons for cosmetic failures are: volume of resection (lumpectomy, quadrant-ectomy), postoperative radiotherapy, deviation from normal breast morphology and tumor location. There are a lot of questionnaires for quality of live assessment. For Bulgarian population N. Mashonov created and adopted original questionnaire. The author investigated quality of live among patients with hard failure during recovery period. PMID- 16044876 TI - [Clotting induction in the mesenterium and erosive stomach mucosa in wistar rats as the result of a catenoidal ultrasonic transducer]. AB - The ultrasonic transducers and instruments used in surgery operate in the range of 1-10 MHz and release significant heat in the surroundings. They need additional cooling system which significantly raises the cost and impedes the equipment operation. Hemostasis can be successfully achieved by employing ultrasonic transducers that operate in the kilohertz range, avoiding the occurrence of inadmissible tissue overheat outside the area of the transducer performance. The paper presents results of the approbation of the effect of a periodical acoustic signal with frequency of 60 kHz over the erosive stomach mucosa and mesenterium of 28 rats, "Wistar" breed, and the period of tissue exposure is 1-10 min. A transducer-catenoidal semi-wave concentrator that provides maximum emitted power of the ultrasonic wave is used and thermal effects are avoided. It is proved that the use of a powerful and localized ultrasonic signal results in erythrocyte diapedesis and forms microhaemorrhagies together with an out-vessel coagulation in the rat stomach mucosa The tissue damages are strictly limited within the area of the emitter contact and they are subject to control during the ultrasonic performance. PMID- 16044877 TI - [Laparoscopic treatment of Morgagni diaphragmatic hernia]. AB - Morgagni-Larey diaphragmatic hernias are very rare. The defect of diaphragm is usually small and the disease passes asymptomatically. If complaints are present and surgical treatment is needed, two types of operative access could be performed--thoracic and abdominal. A 18 years old man with asymptomatic flow of the disease is presented. The laparoscopic treatment was successful. The possible aspects of operative technique are discussed. PMID- 16044878 TI - [A case of a 47 years old hydatid cyst of the liver]. AB - This is a presentation of a clinical observation of a hydatid cyst of the liver, accidentally diagnosed 47 years before the first complains. There were no calcifications on the X-rays before the operation, besides the long duration of its existence. There were no complications before the surgery, and the operation and postoperative period were smooth. The case is presented, mainly because of its extraordinary long period of existence of the parasite in the liver, without any symptoms. This shows, that the processes of aging of the hydatid cyst in the liver, under certain circumstances, can be very slow and asymptomatic. Two of these conditions are intrahepatic (intraparenchymal) localization, and the lack of adjacent large biliary ducts. PMID- 16044879 TI - [Synchronous tumors of the colon]. AB - The synchronous tumors in the colorectal area are cases of particular interest. They are not rarely met and can be omitted, which is a mistake in the process of treatment and diagnostics. Three cases of successfully treated synchronous tumors are presented and conclusions are drawn regarding the diagnostics and behaviour with this type of pathology. PMID- 16044880 TI - [A case of metachronous carcinoma of the colon diagnosed and radical operated with restorative anastomosis after more than 5 years from the first left hemicolectomy]. AB - The authors described their observation about the female patient with an advanced carcinoma (T4) of the descendent colon where left hemicolectomy was done in the beginning of 1998. The patient is operated again after more than 5 years because of the cancer in the site of the anastomosis. Radical reresection with restorative anasomosis is performed and for this reason the colon is putting via mesenteric radix. The case is analyzed and there are suggestion about is this a local recurrence or "de novo" carcinoma It is important to follow up the patients during lifetime for early uncovering of local recurrence. Endocoloscopy and scener are preferable for this aim. PMID- 16044881 TI - [A case of two simultaneous carcinoma lesions of the colon and rectum with different differentiation and stage of neoplastic process]. AB - It considers a patient with simuitant colorectal carcinoma. Both tumor lesions are from different stage and different histological type. The grade of invasion of tumor process does not correspond to the histological type of neoplasma and nider to the size of the primer tumor. The different evolution of those simuitants lesions does not correspond with criteria for histological type and with the stage of primer tumor as a prognostic factors. PMID- 16044882 TI - New legislative efforts put onus on employers. PMID- 16044883 TI - MMA formulary design requires health plans to control costs. PMID- 16044884 TI - Compensation monitor. Quick results, not self-referrals, fuel increase in imaging tests. PMID- 16044885 TI - Back to the future. The re-emergence of the primary care physician. PMID- 16044886 TI - UnitedHealth stakes claims on new drug safety program. PMID- 16044887 TI - Giving more than a nod to the wave of the future. PMID- 16044888 TI - It's a small world after all. Outsourcing makes inroads. PMID- 16044889 TI - Consumer-directed and home-brewed. PMID- 16044890 TI - Consumer-directed health plans: enrollee views, early employer experience. AB - PURPOSE: Consumer-directed health plans (CDHPs) are a new health insurance product that is of growing interest to employers who are struggling to cope with rising health insurance premium costs and to consumers who are desiring more choice and engagement in their health care. This paper presents the results of a study of California consumer awareness of, and attitudes toward, CDHPs in the context of several national surveys and the experiences of some early-adopting employers. DESIGN AND METHODOLOGY: California Health Decisions conducted a telephone survey of 800 insured adult California residents in November 2002. PRINCIPAL FINDINGS: Few respondents had heard of CDHPs. They appealed more to younger, single, less educated, and healthier respondents and those who did not understand them well. The most attractive CDHP features were greater provider choice and health savings accounts' portability and flexibility. Concerns centered on personal financial exposure. CONCLUSION: While CDHPs' commercial market penetration is increasing, their greatest potential future contributions might be to reduce the number of uninsured Americans by offering an affordable health insurance product and to fund additional health services for retirees. As CDHPs further evolve, more consumer involvement in their refinement, implementation, and evaluation is essential. PMID- 16044891 TI - The formulary files. PBMs get thumbs up from satisfied customers. PMID- 16044892 TI - Spinal disc technology seeks to replace body's engineering marvel. PMID- 16044893 TI - Managed care outlook. Things look rosy for large insurers. PMID- 16044894 TI - Improving quality by encouraging providers to use pediatric combination vaccines. AB - In the last 4 decades, the number of diseases that vaccines can prevent has quadrupled, and, correspondingly, so has the number of immunizations that is to be administered before a child's second birthday. Based on current recommendations, a child may receive as many as six vaccine injections in a single visit. Vaccine technology has advanced significantly since the introduction of the first combination vaccine, diphtheria-tetanus (DT), in the 1950s. In the United States today, as many as five antigens can be administered in a single injection, and additional combination vaccines are in the pipeline. Increasing the number of antigens delivered with a single injection minimizes physical discomfort for the child, reduces associated stress for the parent, saves time for the provider, and is likely to improve vaccine coverage and timeliness of administration rates. While national immunization guidelines from the Centers for Disease Control and Prevention call for use of combination vaccines where available, provider and parent perceptions can act as barriers to their optimal use. Managed care organizations (MCOs) have the opportunity to improve quality of care and immunization rates by educating providers on the use of combination vaccines in accordance with the national guidelines. This article examines the evidence for pediatric combination vaccines, discusses barriers to their use among parents and providers, presents quality and cost implications of a managed care policy to broaden their use, and suggests ways in which MCOs can more actively promote appropriate use of combination vaccines by providers. PMID- 16044895 TI - [Developmental dysphasia]. AB - Developmental dysphasia is a specific, primary and lasting oral language disorder (expressive or comprehensive) is the absence of any sensorineural damage oral organ dystunction or psychiatric and psychologic disorders. Pathogensis is still unknown. Diagnosis is based on exprssive and comprehensive language investigation. Therapy should be early and multidisciplinary. PMID- 16044896 TI - [Drug tendon disorders]. AB - Tendinitis is a rare adverse reaction described after drugs use. It's described with drug belonging to the same class. Physiopathology is still unknown. Fluoroquinolones especially pefloxacine, were the most incriminated. This drug induced tendinitis in older people aged more than 60 years. Approximatively one case out of five leads to tendon breaking off. The others lead to a favourable outcome after drug withdrawal. In three cases, tendinitis was described with statines, and concerned 2 men and 1 woman, aged more than 50 years. Simvastatine was suspected in two cases and the outcome was favourable in all cases after drug withdrawal. Other drugs, like corticosteroids, can exceptionally induce this side effect that can be serious. Consequences of drug tendinitis can then be dangerous and prescription of this class, and especially fluoroquinolones, must be cautions. PMID- 16044897 TI - [About a short series of Swanson arthroplasty]. AB - The authors report a series of 12 patients who underwent digital arthroplasty using a swanson implant. Six patients had the sequellae of an injured hand, and the other six patients had rheumatoid arthritis. The majority of patients were men with an average age of 33 years. The patients were examined again about 4 years later on average. The lateral stability of the operated finger was considered to be satisfactory in 11 cases. The mean movement gain was 40 degrees. There was no sign of silicone particle synovitis, because it was fixed arthroplasty. PMID- 16044898 TI - [Knowledge, attitude and behavior about the inherited predisposition to cancer. A survery of a population without any history of cancer in central Tunisia]. AB - The genetic bases of inherited predisposition to cancer are now established. The aim of our study is to value the knowledge, attitude and behavior of the general population about the inherited predisposition to cancer. Our study involved a population of 200 individuals. Without any history of cancer. The mean age of our population was 37.5 years (18 to 74 years). The education level was low in 62.5% of cases (illiterate or primary education). About knowledge: heredity was considered a predisposing factor to cancer by 42.5% of the respondents. About attitude: we noted a cancerophobia in 82% of cases. 86.5% of our respondents trought that an early diagnosis increased the chance of recovery. About behavior: 72.5% of the studied population wishd to know if they were predisposed to develop cancer. In case of pregnancy, 79% wished to know if the foetus wasa cancer gene predisposition carrier. 28% would keep this foetus in case of positive genetic testing. These results are encouraging to develop oncogenetic counselling in Tunisia. PMID- 16044899 TI - Prevalence and severity of asthma, allergic rhinoconjunctivitis and atopic eczema in "Grand Tunis" schoolchildren: ISAAC. AB - The aim of this study was to assess the prevalence and severity of asthma, allergic rhinoconjunctivitis and atopic eczema in 13-14 year old children living in "Grand Tunis". Using the international study of asthma and allergies in childhood (ISAAC) questionnaire, 3350 schoolchildren aged 13-14 years, from the Grand Tunis (Ariana, Ben Arous, Manouba, Tunis) were studied. Our results showed that in the past year 13.2% of children had wheezed, 1.4% had more than 12 attacks and 4.3% had experienced a speech limiting attack. 29.7% had symptoms of allergic rhinoconjunctivitis and 8.3% atopic eczema. The classic preponderance of asthma in boys has not been retrieved in our study. The highest level of wheeze was found in an agricultural area. According the published data, asthma prevalence in Tunisian schoolchildren is intermediate and allergic diseases are perhaps a common childhood diseases in Tunisia. PMID- 16044900 TI - [Diagnostic contribution of HBME-1 and anti-cytokeratin-19 antibodies in thyroid pathology: a retrospective study of 163 cases]. AB - The objective of our study was to estimate the expression of 2 antibodies HBME-1 and anti-cytokeratin-19 and their diagnostic importance in thyroid pathology. MATERIAL AND METHODS: 163 thyroid lesions were retrospectively examined by immnohistochemistry. RESULTS: 92% (46/50 cases) of papillary carcinomas expressed HBME-1 as well as 50% (8/16 cases) of follicular carcinomas and 15% (6/40 cases) of follicular adenomas. 8 insular carcinomas, 5 anaplastic carcinomas, 20 cases of Basedow disease and lymphocytic thyroiditis, and the 24 cases of nodular goiters did not express it or very focally. Anti-cytokeratine-19 marked 92% of papillary carcinomas, 56.2% of follicular carcinomas, 100% of the medullar carcinomas and 45% of follicular adenomas. Whereas the cases of anaplastic carcinomas, Basedow disease, thyroiditis and the cases of nodular goiters were negative or focally marked. CONCLUSION: HBME-1 is an excellent marker for papillary carcinoma which can be helpful in the diagnosis of its follicular variant; the association with anti-cytokeratine-19 increases its specificity. PMID- 16044901 TI - [Functional disability indexes: translation difficulties and cross cultural adaptation problems]. AB - OBJECTIVES: To summarize the difficulties involved in translating tests in Arabic and to describe the translation methods and to apply those to functional indexes. METHOD: Four functional indexes were translated and then subjected to the following test validation methods: back translation, pre-test, and review by an expert committee. RESULTS: Translation problems were underlined. These include in particular the different types of equivalence between the source language and the target language (semantics, idioms, conceptual... equivalences). Problems related to comprehensive literal words were the most observed. CONCLUSION: The current method combining translation with back translation is not sufficient and must be used with, a pre-test and a review committee. PMID- 16044902 TI - [Prevalence of the microscopic colitis to the course of the chronic diarrhea: about 150 cases]. AB - The goal of our survey is to value the microscopic colitis frequency at 150 patients, colliged on a period of 6 years (1997-2003), that have a chronic diarrhea with a normal total coloscopy and that benefitted of systematic biopsies. The microscopic colitis has been recovered at 44 patients on 150 either 29.3%. 16 of our patients (37%) are put on Salazopyrine, they answered well to the treatment with a receding of 3 years. The microscopic colitis is a pathology whose frequency seems under valued, it is certainly benign but can give a frequent diarrhea requiring a curative treatment. PMID- 16044903 TI - Primary myxoid leiomyosarcoma of the ovary. A case report with review of the literature. AB - A case of primary myxoid leiomyosarcoma of the ovary in a 50-year-old Tunisian woman is presented. Bilateral salpingooophorectomy and hysterectomy were carried out without any adjuvant therapy. The tumour were investigated histologically and immunohistochemically. Smooth-muscle actin and progesterone receptors was strongly demonstrated in neoplastic cells, bcl-2 was weakly and diffusely demonstrated. Relevant literature is reviewed based on the histologic and immunohistochemical features with emphasis on diagnosis and therapeutic problems and prognosis indicators. PMID- 16044904 TI - [Endobronchial hamartoma revealed by hemoptysis]. AB - Hamartoma is the most frequent benign tumor of the lung. Its endo bronchial location is rare. We report two cases of endo bronchial hamartoma occurring in 2 men aged 68 and 60 years respectively. The two cases were revealed by hemoptysis. Bronchial fibroscopy showed a bud respectively in the left stump and in the lower left bronchus. Treatment consisted in a pneumonectomy and a lower lobectomy. A histological examination confirmed the diagnosis of endo bronchial hamartoma. CONCLUSION: Diagnosis of endobronchial hamartoma before surgery is difficult. Pulmonary resections are often necessary because of parenchyma lelions caused bronchial obstruction. PMID- 16044905 TI - [Alpha interferon in children with Philadelphia chromosome-positive chronic myeloid-leukaemia]. AB - The present work focuses on the therapeutic efficacy and the toxicity of alpha interferon in patients younger than age 18 years. 5 patients younger than 18 years were treated and followed up between 1990 and 1999 at the department of haematology (Aziza Othmana Hospital) Hydroxyurea was given as initial treatment to all patients. After a median period of 8 months, these patients received alpha interferon (5 millions units/m2 once). Six months after the beginning of the alpha interferon a complete hematologic response was obtained in all patients. The median overall survival was of 66 months: 3 patients are still alive (2 patients in an advanced stage and one patient in chronic phase) and 2 patients died after transformation. The most common reported side effects of alpha interferon were asthenia, weight loss, fever, myalgia, chills and headaches- these toxic manifestations were mild and were noticed in all our patients. Myelosuppression was noted in two patients. Interferon is well tolerated in patients younger than age years 18 old, with CML. It may offer an alternative to bone marrow transplantation in children in the chronic phase of CML without histocompatible donor. The role of new agents such as STI 571 needs to be evaluated as well. PMID- 16044906 TI - [Cardiac amyloidosis revealing multiple myeloma]. AB - INTRODUCTION: Amyloidosis is a rare disease characterized by an extracellular accumulation of a protein polysaccharid complex (Amyloid). Cardiac involvement is considered as a major prognostic factor. OBSERVATIONS: We report the case of two women, hospitalized for heart failure. The diagnosis of cardiac amyloidosis was suggested by echocardiography: Left ventricular concentric hypertrophy and typical amyeloid infiltration with hyperechoic, shiny and granite-like aspect of the interventricular septum. The histological confirmation was obtained by gastric biopsy in the first case and biopsy of the salivary glands in the second revealing an amyloidosis AL. This cardiac amyloidosis was secondary to multiple myeloma: monoclonal Gammopathy with immunoglobulin Lambda in the first and Kappa in the second, and the presence of a plasmocyte infiltration in the sternal puncture. CONCLUSION: Amyloidosis is a rare pathology, the cardiac involvement is frequent in the type AL and can occur with or without clinical manifestations. Echocardiography should be systematic in patients with confirmed amyloidosis. PMID- 16044907 TI - [Ocular findings in megaloblastic anemia associated with diabetes. A case report]. AB - Ocular involvements of Biermer's anaemia are rarely reported in literature. We present a case of Biermer's anaemia associated with diabetes. Ocular examination showed important conjinctival paleness, diffuse retinal ischemia, Roth's tasks, macular oedema and ischemic optic neuropathy. The patient was treated with vitamin B12 intramusculary. A month later, on examination, we noted a regression of optic neuropathy, the aggravation of ischemic retinopathy and persistence of macular oedema. The patient was treated with laser photocoagulation. The majority of ocular manifestations are reversible if treatment is underlaken early. The combination of diabetes with Biermer's anemia deteriorates the ischemic retinopathy and aggavates its prognosis. PMID- 16044908 TI - [Vesico-sigmoid fistula secondary to diverticulitis: a case report]. AB - Vesico-sigmoid fistula is a rare complication of colic diverticular disease. It develops when the bladder sticks to an inflammatory colon making of a communication between the bladder and the digestive segment, usually the sigmoid. Liquid usually passes from the colon to the bladder because of the existing pressure gradient. Hence, urinary symptoms are the most frequent. Surgery is the treatment of choice. The present case is about a 72-year-old man who had a total hematuria for 3 months. The diagnosis of a vesico-sigmoid fistula secondary to colic diverticulitis was established by cystoscopy, colonoscopy and cystography. Surgery was underlaken and the postoperative outcome was excellent. Through this case the clinical features, special investigations and treatment of vesico sigmoid fistula, are reported. PMID- 16044909 TI - [Intestinal T cell lymphoma in coeliac disease. A case report]. AB - Primary intestinal T-cell lymphomas account for about 5% of all primary gastrointestinal lymphomas and are mostly associated with coeliac disease. Intestinal lymphomas are usually discovered during the 4th to 5th decade and constitute an exceptional complication of coeliac disease. This lymphoma is generally a high grade pleomorphic cell lymphoma with large cells. Our case concerned a 30 year old woman with a history of coeliac disease who developed a gastro-enteromesenteric lymphoma. Anatomopathological and immunohistochemical analysis showed an enteropathy associated T-cell lymphoma. We suggest that patients with coeliac disease unresponsive to a gluten free diet or with deteriorating clinical condition be investigated for complications such as enteropathy associated T-cell lymphoma. It is also important to avoid this complication by detecting asymptomatic form of coeliac disease that needs a gluten-free diet. PMID- 16044910 TI - [Effect of HAI-1 over-expression on in vitro growth and migration/motility of SW620 cells]. AB - Hepatocyte growth factor activator inhibitor type 1 (HAI-1) is an effective inhibitor for hepatocyte growth factor activator (HGFA) and serine protease Matriptase, and is involved in HGF/c-Met signaling pathway by regulating activities of HGFA and Matriptase. In an attempt to elucidate the roles HAI-1 play in tumor cell growth and migration, we subcloned full length cDNA of human HAI-1 into mammalian expression vector pcDNA3.1 (+), and transfected the construct into human colorectal cancer cell SW620. The expression of HAI-1 in transfected SW620 cells was demonstrated by Western blot. Growth curve determination, soft-agar colony formation assay, trans-membrane migration assay and wound-healing cell motility assay were used to evaluate the effects of over expression of HAI-1 on cell growth and migration/motility. Both of the growth curve and soft-agar colony formation assays showed that there were no significant differences of growth of HAI-1-transfected cells when compared with the control groups. Trans-membrane migration assay and wound-healing motility assay showed significant inhibition of cell migration/motility ability by the over-expressed HAI-1. Our data suggest that over-expression of HAI-1 in SW620 cells has weak effect on cell growth in vitro, but can significantly inhibit cell migration/motility. PMID- 16044911 TI - [Effect of white spot syndrome virus envelope protein Vp28 expressed in silkworm (Bombyx mori) pupae on disease resistence in Procambarus clarkii]. AB - The vaccine made of recombinant envelope protein (rVp28) of white spot syndrome virus (WSSV) expressed in silkworm (Bombyx mori) pupae using a baculovirus vector was used to investigate the efficacy of oral administration on WSSV disease resistance of Procambarus clarkii. Vaccine was mixed with diet at a ratio of 2% (w/w), and Procambarus clarkii were orally administered throughout 75 days. Vaccination with rVP28 showed the significantly higher cumulative survival compared with positive and negative control (P < 0.05) following an oral challenge on the 35th day post-vaccination (dpv), with PRP values 54.16% and 59.26%, respectively. rVP28 induced higher resistance via IM (intramuscular) injection challenge with WSSV stock, with PRP value of 46.12% and 49.99%, respectively. The survivors were subsequently re-challenged on the 55th dpv. rVP28 induced the significantly higher resistance to oral re-challenge (P < 0.05), with both PRP values 55.80% and 63.16%, respectively. rVP28 induced higher resistance to IM injection re-challenge, with both PRP values 31.25%. A DIG labeled WSSV DNA probe was used to detect WSSV by in situ hybridization. The positive cells were observed in epithelial cells of stomach, hepatopancreas and gut of the infected control crayfish, while negative reaction were observed in the tissues of survivors-vaccinated. These results indicated that vaccination of crayfish with recombinant protein had significant effect on oral infection, and had higher resistance against intramuscular injection challenge. This suggested the protection against WSSV could be induced in crayfish by recombinant protein rVp28 expressed in silkworm pupae. PMID- 16044912 TI - [The mechanism of the recipient maternal-fetal immuno-tolerance induced by the transferred paternal antigen-tolerant T cells]. AB - To study the effect of adoptive transfer of paternal antigen-tolerant T cells on recipient reactive T cells, CBA/JxDBA/2 mating was recruited as an abortion-prone model, and CBA/JxBALB/c mating as a successful pregnancy model. The abortion prone CBA/J females mated with DBA/2 males were injected intraperitoneally with rat anti-mouse CD80 and CD86 mAb or rat isotype IgG at day 4 after gestation (time of implantation). The purified T cells were obtained from spleen of the pregnant CBA/J mice using magnetic beads at day 9 after gestation and labeled with CFSE in vitro. The CFSE-labeled T cells were intravenously injected into other CBA/J females mated with DBA/2 males at day 4 after gestation. The proliferation of recipient splenocytes in response to DBA/2 stimulator cells was evaluated at day 9 after gestation in vitro, and the expressions of intracellular cytokines and costimulatory molecules in CFSE +/- T cells were analyzed by flow cytometry. The results showed that adoptive transfer of either paternal antigen tolerant T cells or T cells from BALB/c-mated CBA/J mice significantly suppressed the proliferation of recipient splenocytes in response to DBA/2 stimulator cells and resulted in lower frequency of cells positive for IL-2, IFN-gamma, CD28 and higher frequency of IL-10,CTLA-4-producing cells in both CFSE+ CD3+ population and CFSE- CD3+ population compared with adoptive transfer of T cells from isotype IgG-treated CBA/J mice, whereas the frequency of IL-4-producing cells did not appear significant change. Our findings suggest that paternal antigen-tolerant T cells transferred in recipient not only function as antigen-specific suppresser cells but also disable the recipient reactive T cells, which co-suppresses maternal rejection to the allogeneic fetus, thus resulting in the decrease of the embryo resorption rate of the abortion-prone mice to that of the normal pregnancy mice. PMID- 16044913 TI - [Cyclosporin a induces titin expression in human trophoblast cells through the MEK/ERK1/2 signal pathway]. AB - To investigate the role of MEK/ERK1/2 signal pathway in the regulation of cyclosporin A(CsA) -induced titin expression in human trophoblast cells. With RT PCR and Western Blot, We examined the titin expression level of human trophoblast cells treated with different concentrations of CsA for various duration, then detected total ERK1/2 and phosphorated ERK1/2 level with Western Blot, and observed effect of U0126 on transcription of titin mRNA in human trophoblast cells stimulated by cyclosporin A. It was found that CsA could activate ERK1/2 in time-dependent and dosage-dependent manner, and induced titin to be expressed in human trophoblast cells. U0126, a MEK inhibitor, inhibited the transcription of titin in dosage-dependent manner. These results indicated that MEK/ERK1/2 signal pathway may play an important role in the expression of titin in human trophoblast induced by cyclosporin A. PMID- 16044914 TI - [Intrathecal injection of muscarinic receptors or GDNF antisense oligonucleotides inhibits the increase of c-Fos expression in locus coeruleus of morphine withdrawal rats]. AB - The antisense approach and immunohistochemistry were used to study the effects of different muscarinic receptor (M) subtypes and glial cell derived neurotrophic factor (GDNF) on the scores of morphine-withdrawal syndrome and the expression of c-Fos in locus coeruleus (LC). Intrathecal injection of M2 receptor antisense oligonucleotides (M2AS-oligo) or GDNF antisense oligonucleotides (GDNFAS-oligo) decreased the scores of morphine withdrawal syndrome. The expression of c-Fos positive neurons in the LC increased in morphine-dependent rats and increased to a greater extent after the injection of naloxone (4mg/kg, ip) in morphine dependent rats. Intrathecal injection of M2AS-oligo or GDNFAS-oligo inhibited the increase of c-Fos expression in LC during morphine withdrawal, but there was no effect in case of M1AS-oligo. The results suggest that M2 receptor of spinal cord mediates the neural activation of LC during morphine withdrawal. And the interaction between neurons and glial cells may be involved in the ascending activation process. PMID- 16044915 TI - [Creation and hybrid identification of the sexual progenies using citrus somatic hybrid as pollen parent]. AB - 110 plantlets were obtained from the cross between citrus allotetraploid somatic hybrid [Hamlin sweet orange (Citrus sinensis Osback) + Rough Lemon (C. jambhiri Luss)] (as the pollen parent) and mono-embryonic diploid type Iben No.4 [Huanongbendizao tangerine (C. reticulata Blanco) x Ichang papeda (C. ichangensis Swingle)] by the means of young embryo culture in vitro, and there were 93 triploids and 17 diploids through chromosome counting and the ploidy analyser identification. RAPD analyses showed that all sexual progenies were hybrid. PMID- 16044916 TI - [Identification of molecular markers linked to the fertility restoring gene for the CMS Capsicum annuum L]. AB - Bulked segregant analysis method was used to identify random amplified polymorphic DNA (RAPD) markers linked to the fertility restoring gene for the cytoplasmic male sterility (CMS) capsicum annum L. Totally 336 random primers were screened on the DNA samples of restorer and sterile bulks. Primer S418 produced a special band in restorer line. It was about 3000 bp, including two fragments 1515 bp and 1162 bp. Fluorescence in situ hybridization(FISH) indicated the fragment of 1515 bp only existed in restorer line.It was designed to S418(1515). Analysis of the sequence indicated S418(1515) was unknown before. The homology of blastn was less than 40%, however the homology of tBlastx indicated this sequence was high homologous with the part sequences of rice which were distributed on 2,4,7,10 chromosomes. It suggested this sequence might have the similar function with them. This result offered a good foundation to research the molecular mechanism of fertility restoration for CMS capsicum. Based on the sequence, special primers were designed to transform the RAPD marker to PCR marker. The result indicated that these primers could be used to screen the restorer lines from a large quantitive of candidate lines. PMID- 16044917 TI - [Study on genetic instability of NM23H1 gene in Chinese with the epithelial ovarian tumor]. AB - The aim of this study was to examine microsatellite instability (MSI) and loss of heterozygosity (LOH) of locus D17S396 on chromosome 17 and their influence on the expression of nm23H1 in the epithelial ovarian tumors, which may provide experimental basis for the mechanism of nm23H1 gene and tumor metastasis. Techniques such as DNA extraction from formalin-fixed paraffin-embedded tissues, polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP), ordinary silver stain were used to study MSI and LOH of locus D17S396. Envision immunohistochemistry and Leica-Qwin computer imaging techniques were used to assess the expression of nm23H1 gene. In our experiments, the frequency of heredity instability of malignant ovarian tumors was 40%, which is higher than that of borderline ovarian tumors, while there were no heredity instability occurred in benign ovarian tumors and normal ovarian tissue. Among 25 epithelial ovarian carcinomas, the frequency of LOH in lymph node metastasis cases (66.67%) was significantly higher than those without metastasis (10.53%). Moreover, the frequency of LOH was higher in FIGO stage III and IV than in stage I and II. However, the frequency of MSI showed no correlation with any clinicopathologic characteristics. The positive frequency of nm23H1 protein in the ovarian epithelial carcinoma and borderline tumors were 56.00% and 57.14%, respectively. They were both higher than those of the benign tumors and normal ovarian tissue. In the epithelial ovarian carcinomas, the positive frequency of nm23H1 protein in lymph node metastasis case was significantly lower than those without metastasis. FIGO stage III and IV also exhibited lower positive frequency of nm23H1 protein compared with stage I and II. Furthermore, there was no difference in nm23H1 protein expression intensity analyzed by computer imaging. In the epithelial ovarian carcinomas, the positive frequency of nm23H1 protein in LOH positive group was 0.00%, which is lower than that of LOH negative group (P < 0.01). The results indicated that the heredity instability of nm23H1 gene might be implicated in pathogenesis and progression of epithelial ovarian tumor. The occurrence of LOH might be the molecule marker of the deteriorism of ovarian tissue. Both MSI and LOH of nm23H1 gene controlled development of the epithelial ovarian tumor independently in different paths. LOH could inhibit the expression of nm23H1 in local tissue of the epithelial ovarian carcinoma, which endowed it with high aggressive and poor prognosis. Increasing the amount of nm23H1 protein expression could effectively restrain metastasis of the ovarian epithelial carcinoma and improve prognosis of patients. PMID- 16044918 TI - [Effect on development in NT embryos after transplantation of nuclei derived from transfected goat fetal fibroblasts suffering different treatments into enucleate eggs]. AB - In order to improve the development rate of preimplantation nuclear transfer embryos (NT embryos) after transplanting nuclei derived from transgenic goat fetal cells, the donor fetal fibroblasts starved for 5 days in DMEM containing 0.5% FCS were divided into three groups and treated with different methods respectively before using as donor cell. Group 1 was frozen at -80 degrees C or in liquid nitrogen for several days or months. Group 2 was at first treated as the same as group 1, then cultured for 2-5 days in DMEM containing 10% FCS and starved for another 5 days subsequently. Group 3 was cultured for 2-5 days in DMEM containing 10% FCS and starved for another 5 days subsequently. The rate of G0/G1 phase cells from group 2 was 95.68% and significantly different from group 1's 88.66%. The rate of survival cells from group 2 was 99.9% and significantly different from group 1's 80.00% (P < 0.05).The morula- blastocyst stage NT embryos development rate of group 2 was 66.09% and significantly different from group 1's 22.00% and group 3's 50.51% (P < 0.05). All NT embryos of above three groups were transferred into synchronous oestrus recipients and the pregnant status of recipients was checked by B-mode ultrasound diagnosis after 35 days. The recipient pregnancy rate of group 2 was 45.83%, much higher than that of group 1(20.00%) and group 3 (29.58%). The result of this experiment showed that donor cells treated with freezing and two times starvation could significantly improve the rate of G0/G1 phase cells, the rate of survival cells, the NT embryos development rate and the recipient pregnancy rate. PMID- 16044919 TI - [Effective cryopreservation of human embryonic stem cells by programmed freezing]. AB - Cryopreservation of human embryonic stem cells is an important and unsolved problem. A computer-controlled programmable cooler was used in the preservation of ES cells. Several effects have been experimentally studied, which include the cooling rates, the temperature of seeding, the temperatures before the samples being plunged into liquid nitrogen, and the cryoprotective agents. It was found that the favorable constitution of cryoprotective agents was Me2SO+ FBS+DMEM(1:3:6, v/v/v) with cooling protocol of -0.5 degrees C/min from 0 degrees C to -35 degrees C (seeding at -10 degrees C), and being plunged into the liquid nitrogen immediately. The high survival rate (81.8%) was obtained. PMID- 16044920 TI - [Correlation between the expression of VEGF-C mRNA, VEGFR-3, CD31 and tumor metastases in Chinese with prostate cancer]. AB - The expressions of VEGF-C mRNA, VEGFR-3 and CD31 were studied in order to investigate the correlation between them and neoangiogenesis, hyperplasia of micro-lymphatics and tumor metastases. 34 cases of prostate cancer tissue and 12 cases of adjacent nontumorous tissue specimens were tested. They were marked by VEGFR-3 and CD31 with immunohistochemistic staining and analyzed with image, the micro-lymphatics count (MLC) and microvessel density (MVD) were counted using Weidner's highest vessel density count method; the expression of VEGF-C mRNA was inspected in situ hybridization. The expression of VEGF-C mRNA was 44.12% positive in 34 cases of prostate cancer, the MLC (8.26 +/- 2.73)mm2 and MVD (74.82 +/- 11.76)mm2 in prostate cancer were significantly higher than those in adjacent nontumorous tissue (MLC, 4.82 +/- 3.48/mm2; MVD, 32.86 +/- 5.41/mm2). In addition, there was a correlation between the expression of VEGF-C mRNA and micro lymphatics metastases and there was a positive correlation between the expression of VEGFR-3 and CD31. The expressions of VEGF-C mRNA , MLC, MVD in stage III and IV and those who have lymph metastasis were higher than those in stage I and II and those who have no lymph metastasis; the expressions of VEGFR-3 and CD31 in VEGF-C mRNA positive groups were significantly higher than those in negative groups. The difference of histopathologic grading in prostate cancer had no statistical significance. VEGF-C could accelerate the hyperplasia of micro lymphatics and neoangiogenesis induced by tumor and play an important role in tumor lymph metastases. There was a close correlation between the expressions of VEGFR-3, CD31 and tumor metastases. The increase of MLC and MVD on prostate cancer indicated the hyperplasia of new micro-lymphatics and neoangiogenesis in the tumor tissue, which could also be a signal to determine the tumor metastases in clinic. PMID- 16044921 TI - [Effects of anther--pretreatments on the survival percentage of isolated microspores of maize]. AB - Using maize anthers of 2 cultivars as experimental materials, the effects of mannitol, colchcine and proline concentration in pretreatment solution and pretreating temperature on the survival percentage of isolated microspore were investigated. The survival percentage of isolated microspores were able to be respectively and obviously improved by pretreating anthers with mannitol from low to high concentration (0.35 mol/L, 24h --> 0.40 mol/L, 24h --> 0.45 mol/L, 24h), or with colchicine 200 mg/L or with proline 200 mg/L and pretreating at 15 degrees C for 48h. The probable mechanism of pretreatments was discussed. PMID- 16044922 TI - [Mapping of apple Co gene using SSR markers]. AB - Columnar apple is an important genetic resource for tree form breeding of apple. In this study, 106 individuals of the F1 population derived from 'Spur Fuji' (coco)x 'Telamon'(Coco) were used as plant materials for screening SSR markers linked to gene. By bulked segregating analysis (BSA), eight SSR markers from the tenth linkage group of apple genome were tested. Finally, three of them, COL, CH02a10 and CH03d11, were identified to be SSR markers of Co gene. Linkage analysis showed that the genetic distance of COL, CH02a10 and CH03d11 to Co locus was 15.3cM, 22.2cM and 3.9 cM, respectively. On the linkage map of these markers, Co gene was located between COL and CH03d11. PMID- 16044923 TI - The times they are A-changin'. PMID- 16044924 TI - The effect of malnutrition on morbidity after Surgery for chronic pancreatitis. AB - Protein-energy malnutrition is a notable problem in the management of patients with chronic pancreatitis. The effect of malnutrition on pancreatic surgery is not well known. The records of 313 consecutive patients who underwent lateral pancreaticojejunostomy (LPJ, n = 152), pancreaticoduodenectomy (PD, n = 78), or distal pancreatectomy (DP, n = 83) for chronic pancreatitis were retrospectively reviewed and analyzed. Subjective Global Assessment, Nutritional Risk Index, and Instant Nutritional Assessment were used to assess the nutritional state. An average of all three nutritional indexes was established, and patients were categorized into well nourished (n = 101) as well as mild (n = 91), moderate (n = 94), and severe malnourished (n = 27). Poor nutritional state was associated with an increase in postoperative complication rate (LPJ: well nourished 14%, mild 25%, moderate 31%*, severe 50%*; PD: well nourished 44%, mild 44%, moderate 60%, severe 88%*; DP: well nourished 17%, mild 13%, moderate 30%, severe 55%*; *P < 0.045 vs well nourished). Low serum albumin levels also increased operative morbidity. The increase in morbidity was reflected by a higher rate of infectious complications as well as increased ICU stay. Body mass index and weight loss did not contribute to change in outcome. Malnutrition was associated with a higher incidence of postoperative complications after surgery for chronic pancreatitis. An increase in operative morbidity might be related to decreased protein synthesis and impaired immunocompetence. PMID- 16044925 TI - Comparison of results in endovascular interventions for infrainguinal lesions: claudication versus critical limb ischemia. AB - This study analyzed clinical success, patency, and limb salvage after endovascular repair in patients treated for chronic limb ischemia presenting with claudication versus critical limb ischemia. Between October 2001 and August 2004, 115 patients (mean age 71) underwent endovascular treatment for infrainguinal arterial disease. Techniques included subintimal angioplasty and transluminal angioplasty with or without stents. Lesions were classified according to Transatlantic InterSociety Consensus. Follow-up (mean 11 months) included physical exam, ankle-brachial index, and duplex ultrasound. Patency rates were determined using Kaplan-Meier and compared by log-rank analysis. One hundred ninety-nine lesions were treated in 121 limbs using percutaneous techniques. Comorbidities were similar except higher rates of diabetes mellitus (67% vs 41%, P < 0.001) and chronic renal insufficiency (22% vs 7%, P < 0.05) were found in critical limb ischemia patients. Primary patency for claudicants was 100 per cent, 98 per cent, and 85 per cent at 3, 6, and 12 months and 89 per cent, 80 per cent, and 72 per cent for critical limb ischemia, respectively (P = 0.06). Limb salvage was 91 per cent at 12 months for critical limb ischemia patients. Morbidity was similar between groups, and there was no perioperative mortality. Percutaneous intervention for both claudication and critical limb ischemia provides acceptable 12 month patency with limited morbidity. PMID- 16044926 TI - Prevalence of delayed hemothorax in blunt thoracic trauma. AB - Delayed hemothorax (DHTX) is rarely seen. On an 8-year retrospective analysis of blunt thoracic trauma (BTT), hemothorax (HTX) was diagnosed in 167 patients: 18 children, 113 adults, and 36 elderly. No statistical differences were seen in any age groups regarding Injury Severity Score (mean ISS, 30.54), critical care length of stay (CLOS, 9.0), and hospital LOS (HLOS, 11.21). Mortality rate was 18 per cent in adults and 28 per cent in elderly (P value < 0.0001). HTX was acute in 160 and delayed in 7 patients. Two-thirds of HTX patients were males and 75 per cent had rib fractures. All of our DHTX patients were males (5 adults and 2 elderly) and had rib fractures. Acute HTX was seen in younger patients (43.3 vs 56.1 years, P value 0.46), with higher ISS (31.44 vs 14.43, P value < 0.001), CLOS (7.19 vs 3.0 days, P value 0.511) and HLOS (11.9 vs 11.6, P value 0.468). Mortality was 22.5 per cent in AHTX and none in DHTX. Eighty-six per cent of DHTX and 49 per cent of AHTX patients went home on discharge. DHTX was rare (5%) in the current report with lower ISS, HLOS, and no mortality. Patients with rib fractures should be watched for development of DHTX as timely diagnosis and treatment is essential for favorable outcome. PMID- 16044927 TI - Neoadjuvant chemotherapy in stage III breast cancer. AB - Neoadjuvant chemotherapy in advanced breast cancer can potentially downstage disease prior to definitive surgery. In this study, a doxorubicin-based neoadjuvant regimen was administered to stage III breast cancer patients to assess 1) primary tumor response, 2) tumor involvement of resection margins, and 3) predictive value in cancer outcome. Eighty-two patients with stage IIIA and IIIB breast cancer diagnosed between 1990 and 2003 were studied. All patients received similar chemotherapy regimens, consisting of doxorubicin, cisplatin, and 5-fluorouracil, plus surgery and radiation therapy. End points measured include primary tumor response [complete response (CR) = 100%, partial response (PR) > 50%, or no response (NR) < or = 50%], resection margins for tumor, disease-free, and overall survival. Kaplan-Meier and log-rank tests were performed. Of the 82 patients studied, 34 received neoadjuvant therapy, 48 received conventional postoperative treatment. Seventy-two per cent of the stage IIIB and 22 per cent of the stage IIIA patients received neoadjuvant therapy. In the neoadjuvant group, 29 (85%) patients demonstrated tumor response, 9 (26%) of which were CR. Tumor-free resection margins were achieved in 94 per cent of the neoadjuvant group. Survival analysis demonstrated no benefit comparing neoadjuvant versus postoperative adjuvant therapy but hints at improved disease-free survival in neoadjuvant CR patients (log-rank test, P = 0.07). Eighty-five per cent of patients with stage III breast cancer treated with neoadjuvant chemotherapy experienced clinical response, with 26 per cent CR, and 97 per cent tumor-free resection margins. CR may portend a better cancer outcome. PMID- 16044928 TI - Treatment of isolated penetrating flank trauma. AB - We present our experience during the past 5 years with isolated penetrating flank trauma. We examined the evaluation process, treatment plan, length of stay, outcome, and complications of patients with isolated penetrating injuries to the flank. There were four patient groups: 1) laparotomy (n = 5); 2) local wound exploration (n = 1); 3) laparoscopy (n = 8); and 4) observation and triple contrast CT scan (n = 9). All patients were male. The mean length of stay was 15 days for group 1, one day for group 2, two days in group 3, and 1.5 days in group 4. Three patients had diaphragmatic injuries, which were repaired laparoscopically. One patient with local wound exploration re-presented with a diaphragm hernia. A negative triple contrast CT scan resulted in no subsequent intervention. There were no missed injuries or complications from CT scan + observation or from diagnostic laparoscopy. We believe that laparoscopy may emerge as a useful adjunct in the treatment of patients with flank trauma. We present an algorithm for the management of penetrating flank trauma. PMID- 16044929 TI - Leiomyosarcoma of the inferior vena cava: surgical management and clinical results. AB - Leiomyosarcoma of the inferior vena cava (IVC) is a rare lesion with less than 300 cases reported. Optimal management and long-term outcomes are not well described. From August 1984 to June 2004, eight patients with leiomyosarcoma of the IVC were treated at our institution. Clinical and pathologic data, surgical management, and outcomes were assessed. Eight cases were identified (4 males) with a median age of 52 (range 29-66). Presenting symptoms included abdominal pain (n = 5, 63%), lower extremity edema (n = 2, 25%), and palpable mass (n = 2, 25%). Tumor location was between the renal and iliac veins (low) (n = 4, 50%), between the hepatic and renal veins (middle) (n = 3, 38%), and above the hepatic veins with right atrial extension (high) (n = 1, 12%). Two patients with preoperative IVC occlusion were managed with tumor excision and IVC ligation. Three patients had primary repair of the IVC after tumor excision. A polytetrafluorothylene (PTFE) tube graft was used for IVC reconstruction in three cases. There was no postoperative mortality. Postoperative morbidity included deep venous thrombosis (DVT) (n = 1), lower extremity edema (mild n = 1; moderate n = 1), GI bleed (n = 1), and chronic renal insufficiency (n = 1). One patient is currently receiving adjuvant chemotherapy. Four patients received chemotherapy after recurrence, and one received palliative radiation therapy as well. Median survival to this point was 60 months with a median follow-up of 39 months. The 5 year overall survival and disease-free survival was 31 per cent for both (CI 0.1 1.0). The type of IVC reconstruction had no effect on survival (P = 0.22). Recurrence was discovered in four patients (50%) at a median time of 14 months. Resection of leiomyosarcoma of the IVC should be attempted whenever feasible. The management of the IVC can be managed with primary repair, ligation, or prosthetic graft. Long-term survival is possible if complete resection can be achieved. PMID- 16044930 TI - Absence of the retrohepatic inferior vena cava: what the surgeon should know. AB - Anomalies of the inferior vena cava are rare and can easily be misdiagnosed if one is not aware of such variants. We report a case of a 57-year-old patient that had a percutaneous CT-guided biopsy of what was considered to be enlarged pericaval lymph nodes. This occurred because of the poor technique of the CT scan and the unawareness of such a condition. These changes were related to a proximal varicele of an absent, retrohepatic inferior vena cava. There was an extensive collateral network with filling of large azygos and hemiazygos veins draining through the posterior mediastinum into the superior vena cava. This case emphasizes the importance of correctly identifying vascular anomalies before the initiation of biopsy attempts in order to prevent the risk of major complications that could arise during such biopsy. PMID- 16044931 TI - Acute abdomen secondary to ascaris lumbricoides infestation of the small bowel. AB - Ascariasis is a helminthic infection commonly found in tropical climates. It often propagates in communities of low socioeconomic status secondary to contamination of the soil and water supply with human feces. We present a case report of a 42-year-old Asian-Indian female presenting with a long-standing history of severe recurrent postprandial epigastric pain, requiring multiple hospital admissions. Ultrasound, computed tomography (CT), and nuclear biliary scan were negative. She underwent esophagogastroduodenoscopy that suggested ischemia. Magnetic resonance angiography (MRA) and mesenteric angiography were inconclusive. As conservative treatment had been unsuccessful, a small bowel series was performed. The radiographs demonstrated characteristic findings of Ascaris lumbricoides infestation. Although the prevalence, diagnosis, and subsequent treatment of an acute abdomen secondary to Ascaris lumbricoides infestation is commonly seen in developing countries, clinicians in developed countries may not consider this entity when faced with a patient with similar symptoms. We frequently care for immigrants from developing countries and our own citizens who visit the countries where ascariasis is endemic. Therefore, heightened awareness of Ascaris lumbricoides infection (ALI) presenting as an acute abdomen is necessary. The diagnosis requires an experienced radiologist and knowledge by the clinician of treatment options and of when a surgeon should be involved. PMID- 16044932 TI - Thyroglossal duct cysts: a consideration in adults. AB - Thyroglossal duct cysts (TDC) are one of the most common pediatric midline neck lesions. Although they are present from birth, they usually become symptomatic in early childhood as a mass or draining sinus. Infection and abscess formation are frequent complications due to a communication between the cyst and the mouth with subsequent contamination by oral flora. We present a case of a 37-year-old male who presented with a newly symptomatic thyroglossal duct cyst. The patient presented to the infirmary with pain in the anterior neck particularly with swallowing. A midline mass was visible and palpable on examination. Subsequent neck exploration revealed a thyroglossal duct cyst filled with purulent material. Although uncommon in adults, thyroglossal duct cysts should be a part of the surgeon's differential diagnosis when presented with a neck mass. One should remember that an infected neck mass is the common presentation of thyroglossal duct cysts in adults, and the appropriate diagnostic studies need to be performed in order to best determine the diagnosis. Once diagnosed, the TDC is best treated using the Sistrunk procedure to limit recurrence. PMID- 16044933 TI - The utility and timing of surgical intervention for parapneumonic empyema in the era of video-assisted thoracoscopy. AB - Empyema, a pyogenic or suppurative infection of the pleural space, continues to cause significant morbidity and mortality in patients with pneumonia. The advent of video-assisted thoracoscopy has placed the treatment algorithm of empyema in flux. We retrospectively reviewed all patients who underwent surgical treatment for parapneumonic empyema from January 1, 1999, through December 31, 2003. Data collected included demographic information, preoperative CT scanning/ thoracostomy tube placement, morbidity/mortality, days from admission to surgery, and postoperative length of stay. We compared patients undergoing video-assisted thoracoscopy to those requiring conversion to open thoracotomy and those who had initial open thoracotomy. Morbidity and mortality rates were similar among all groups. Conversion rate to open thoracotomy was 21 per cent. We found patients operated on within 11 days of admission had a shorter postoperative length of stay with similar morbidity and mortality. Our data supports early aggressive surgery treatment for parapneumonic empyema. PMID- 16044934 TI - Endovascular management of ruptured, mycotic abdominal aortic aneurysm. AB - Patients with mycotic aneurysms have a high mortality rate. The standard surgical approach can be exceptionally difficult and fraught with complications. There has been reluctance to insert an endograft into an infected field. We believe that this thought should be challenged and present a case of a successful endovascular repair of a ruptured, mycotic abdominal aortic aneurysm. The patient is a 63-year old man with severe medical comorbidities and methicillin-sensitive Staphylococcus aureus. He required 6 units of red blood cells on admission. Magnetic resonance angiography (MRA) showed a contained rupture of his distal abdominal aorta, and he underwent emergent endovascular repair. An aortomono iliac device (12 mm x 10 cm iliac extension limb) was inserted along with coil embolization of his right common iliac artery and a femoral-femoral bypass. He did not require additional transfusions after the procedure and was discharged in good condition. He is on antibiotics and doing well 1 year post-op. Endovascular management of ruptured, mycotic aneurysms is feasible. In fact, it is an attractive approach for a medically compromised patient subset that would carry an exceptionally high mortality rate with traditional surgical repair. Further follow-up is necessary to determine its long-term efficacy. PMID- 16044935 TI - Blunt trauma to the thyroid: a case report. AB - Thyroid injury is a rare phenomenon in cases of blunt neck trauma. Symptoms are often subtle or not present on initial exam and can be rapidly life-threatening when airway compromise ensues. We describe the case of a 50-year-old woman who developed neck pain and swelling, dysphagia, and hoarseness after a rear-end collision in which she was the restrained driver, hitting her anterior neck against the steering wheel. Neck CT revealed fragmentation and hematoma within the right thyroid lobe. Arteriogram showed no vascular injury to the neck. The patient was observed in the ICU and was discharged home 3 days later without operative intervention. We believe that in the acute setting, a stable traumatic thyroid hematoma without airway encroachment may be safely observed. Increasing size or compromise of airway integrity should be indications for early intubation and neck exploration. PMID- 16044936 TI - Adult gastric duplication cysts: a case report and review of literature. AB - In this report, we present the rare case of a 53-year-old white female with a gastric duplication cyst. Diagnosis was made with a combination of CT scan and endoscopic ultrasound. The cyst was removed by using a seromuscular incision and enucleation. A review of English literature was performed to help define the etiology, workup, and treatment of adult patients. Theories of origin include embryologic enteric diverticula, incomplete notochordal plates separation, and embryonic longitudinal folds that fuse with a bridge. The essential features of duplication are that the cyst wall is contiguous with the stomach, is surrounded by smooth muscle, has a common blood supply, and is lined with epithelium. Few duplication cysts are diagnosed after the age of 12 and in adults present a diagnostic dilemma. In the past, preoperative diagnosis of gastric duplications was the exception, but with advances in imaging, most importantly endoscopic ultrasound, the diagnosis is established before surgery in a majority of the cases. Gastric duplication cysts have been removed using various methods; however, as enucleation can be performed easily with little disruption of normal anatomy, it should be considered the procedure of choice. PMID- 16044937 TI - De Garengeot hernia: appendicitis within a femoral hernia. AB - Many surgeons are familiar with Amyand hernia, which is an inguinal hernia sac containing an appendix. However, few surgeons know of the contribution of Rene Jacques Croissant de Garengeot, an 18th century Parisian surgeon, to hernias. He is quoted in the literature as the first to describe the appendix in a femoral hernia sac. We discuss the case of an 81-year-old woman who presented with appendicitis within a femoral hernia, a rare finding at surgery that is almost never diagnosed preoperatively. We also propose crediting Croissant de Garengeot by naming this condition after him. Although his full last name is Croissant de Garengeot, for convenience we suggest the simple diagnosis of "de Garengeot hernia." PMID- 16044938 TI - Synchronous diverticular perforation: report of a case. AB - Synchronous colonic events are rare. Diverticulitis is the most common and lethal cause of colonic perforation. The first case in the literature of a synchronous diverticular perforation is presented. The patient was admitted with peritonitis. An exploratory laparotomy showed cecal and sigmoid perforations. Resection and primary anastomosis with a protective loop ileostomy was performed. Microscopic evaluation confirmed the presence of cecal and sigmoid perforated diverticuli. Diverticular disease is present in up to two-thirds of patients 80 years of age and older. Right-sided free colonic perforation is rare. Resection and primary anastomosis is feasible in the treatment of perforated diverticular disease. In this report, we emphasize the importance of a thorough abdominal exploration at the time of surgery. PMID- 16044939 TI - Not all inflammation in the right lower quadrant is appendicitis: a case report of Escherichia coli O157:H7 with a review of the literature. AB - Although significant work has been presented on this subject in pediatric, infectious disease, and epidemiologic literature, there is a noteworthy lack of information on Escherichia coli O157:H7 in any surgical journals. As this disease can present with signs and symptoms often ascribed to the acute abdomen, it is imperative that the general surgeon, pediatric surgeon, and colorectal surgeon are all familiar with this infection and its clinical ramifications. A case report followed by a review of the literature is presented. PMID- 16044940 TI - Cryptosporidiosis presenting as acute appendicitis: a case report. AB - Although uncommon in the United States, cryptosporidiosis can be life-threatening in an immunosuppressed host. Rarely, an acute infection of this gastrointestinal illness can present as another disease entity. We present only the third reported case of cryptosporidial infection presenting as acute appendicitis in a 17-year old HIV+ patient. PMID- 16044942 TI - Price survey. Suture makers vie for market. PMID- 16044941 TI - Extreme makeover for Hartford's print shop. PMID- 16044943 TI - Gainsharing: more trouble than it's worth? PMID- 16044944 TI - Mammalian sex hormones in plants. AB - The occurrence of mammalian sex hormones and their physiological role in plants is reviewed. These hormones, such as 17beta-estradiol, androsterone, testosterone or progesterone, were present in 60-80% of the plant species investigated. Enzymes responsible for their biosynthesis and conversion were also found in plants. Treatment of the plants with sex hormones or their precursors influenced plant development: cell divisions, root and shoot growth, embryo growth, flowering, pollen tube growth and callus proliferation. The regulatory abilities of mammalian sex hormones in plants makes possible their use in practice, especially in plant in vitro culture. PMID- 16044945 TI - Characterization of human pituitary adenomas in cell cultures by light and electron microscopic morphology and immunolabeling. AB - The morphology and hormone production of pituitary adenoma cell cultures were compared in order to highlight their characteristic in vitro features. Cell suspensions were prepared from 494 surgical specimens. The 319 viable monolayer cultures were analyzed in detail by light microscopy and immunocytochemistry within two weeks of cultivation. Some cultures were further characterized by scanning, transmission and immunogold electron microscopy. The viability and detailed in vitro morphology of adenoma cells were found to be characteristic for the various types of pituitary tumors. The sparsely granulated growth hormone, the corticotroph and the acidophil stem cell adenomas provided the highest ratio of viable cultures. Occasionally, prolonged maintenance of cells resulted in long term cultures. Furthermore, a variety of particular distributions of different hormone-containing granules were found in several cases. Both light microscopic and ultrastructural analyses proved that the primary cultures of adenoma cells retain their physiological features during in vitro cultivations. Our in vitro findings correlated with the routine histopathological examination. These results prove that monolayer cultures of pituitary adenoma cells can contribute to the correct diagnosis and are valid model systems for various oncological and neuroendocrinological studies. PMID- 16044946 TI - Autocrine growth regulation of W12 and GCA cells in culture. AB - Two rat kidney cell lines transformed by two strains of ASV virus were investigated. It was demonstrated that these two lines (1) showed density independent growth, (2) had a decreased requirement for serum in the culture medium, (3) had the ability to grow in a chemically defined medium (without serum), and the rate of this growth had increased with the increase in starting density of cells, and (4) had the ability of anchrage-independent growth, even without serum. These results confirmed autostimulation of growth of W12 and GCA cells. It was also shown that the crude conditioned media contained autocrine growth factors, which could be extracted with 1M acetic acid. The extracts (AEs) stimulated the growth of the parental cells and NRK-49F cells almost as well as 5% calf serum and the extraction resulted in several-fold purification of mitogenic substances. These substances were not only specific to parental lines, but also stimulated growth of other transformed lines and normal NRK-49F cells. Extracts from the conditioned media of W12 and GCA cells intensified the rate of anchorage-independent growth in the concentration-dependent manner. In AE-W12, two peaks of mitogenic activity were detected (F1, F2) and similarly in AE-GCA (F3, F4). Fractions F2 (approximately 8 kDa), F3 (approximately 25 kDa) and F4 (approximately 12 kDa) were thermostable but F1 (approximately 45 kDa) was thermolabile. All four fractions were sensitive to trypsin and DTT treatment, and were acid-stable. Using ELISA kit it was shown that W12 and GCA cells released TGFbeta1 and GCA cells released very small quantities of bFGF. These results confirmed the autocrine regulation of growth in both cell lines. PMID- 16044947 TI - The effects of synthetic salmon calcitonin on thyroid C and follicular cells in adult female rats. AB - Structural and morphometric features of thyroid C and follicular cells were studied in adult rat females after treatment with synthetic salmon calcitonin (CT). The animals were chronically treated with either a low (10 IU/kg b.w) or a high (100 IU/kg b.w) dose of CT. A stereological method was applied to determine the volume density and the number of immunoreactive C cells. The height and volume density of follicular epithelium, colloid, interstitium and the follicles (epithelium plus colloid), as well as the index of activation rate were calculated. A significant decrease in body weight, as well as the volume density of immunoreactive C cells and the number of C cells per mm2, was observed in rats treated with both doses of CT. The height and volume density of follicular epithelium and follicles, as well as the index of activation rate were significantly increased in the animals given the high CT dose, while the volume densities of colloid and interstitium were reduced. No significant changes in the examined morphometric parameters were detected after treatment with the low CT dose. According to these results it can be concluded that the structural features of thyroid C and follicular cells were affected by the high dose CT treatment in the opposite manner, while the low dose CT treatment influenced only C cells. PMID- 16044948 TI - Maspin and c-erbB-2 expression in correlation with microvessel density in invasive ductal breast cancer. AB - Maspin is a unique member of the serpin family involved in regulation of cell migration, apoptosis and angiogenesis in breast and prostate cancers. In this study maspin expression in comparison with c-erbB-2 (HER2/neu) oncogene expression and microvessel density was investigated. The examined material included specimens of primary invasive ductal breast cancer derived from 69 patients. They were analyzed immunocytochemically to assess maspin and c-erbB-2 expression, as well as microvessel density using endothelium marker CD31. In the studied cancers, maspin expression in cancer cells was detected in more than half of the cases (50.73%). Although statistically insignificant (p=0.27), maspin expression showed decreasing tendency with the increase of tumor grade. C-erbB-2 oncogene expression was observed in 78.26% of the examined cancers. Statistically significant positive correlation was found between c-erbB-2 expression and tumor grade (p<0.005). Analysis of the dependence between maspin and c-erbB-2 expression exhibited statistically significant inverse correlation (p<0.001). Mean microvessel density (MVD) of the studied cancers was 71.64 (SD=19.36). MVD decreased with the increase of maspin expression, whereas in the cases showing c erbB-2 overexpression MVD was clearly higher. Both correlations were statistically significant (p<0.005). In conclusion, it could be stated that increase in maspin expression is associated with weaker expression of c-erbB-2 oncogene and lower microvessel density, which implies a significant role of maspin in tumor biology. However, the exact mechanism of maspin action (including its potential role in angiogenesis), as well as the assessment of its prognostic significance in breast cancer require further studies. PMID- 16044949 TI - Flow cytometric analysis of CD55 and CD59 expression on blood cells in paroxysmal nocturnal haemoglobinuria. AB - PNH is a rare clonal disorder of hematopoietic stem cells, therefore all blood cells lineages are involved. The main feature is an increased sensitivity of erythrocytes to complement-mediated cell lysis due to deficiency of membrane bound GPI (glycosylphosphatidylinositol)-anchored proteins which normally function as inhibitors of reactive hemolysis. In the present study, we performed flow cytometric analysis using monoclonal antibodies against CD55 and CD59 for the detection of PNH-type clone in the blood of 50 patients (28 females and 22 males, age range 7-67 yrs). In one patient only we found a large population (95%) of granulocytes with decreased expression of both CD55 and CD59 molecules (type I PNH) and in two others with partial loss of CD55 expression (type II PNH). The expression was determined chiefly on granulocytes which in the control group showed reliable and high expression of CD55 and CD59. PMID- 16044950 TI - [Drug delivery systems using nano-sized drug carriers]. AB - Nanotechnology has attracted great attention all over the world in recent several years and has led to the establishment of the novel technical field of "nanomedicine" through collaboration with advanced medical technology. Particularly, site-specific drug targeting using particle drug carrier systems has made substantial progress and been actively developed. This review explains the essential factors (size and chemical character) of drug carriers to allow long circulation in the bloodstream avoiding the reticuloendothelial system, and shows the present status and future perspective of several types of nano-carrier systems (water-soluble polymer, liposome and polymeric micelle). We also introduce the novel concept of multi-targeting system (combination of two or more targeting methodologies) for ideal drug therapies. PMID- 16044951 TI - [A historical perspective on familial cancer]. PMID- 16044952 TI - [Genetic medicine in the university hospital]. AB - The importance of genetic medicine is growing along with the development of genome science. Especially for hereditary cancer syndromes, genetic counseling and genetic tests are becoming an essential part of the medical service for those diseases. However, in Japan, there is a shortage of clinical geneticists who are familiar with hereditary tumor syndromes. There are also many other problems such as the cost of genetic tests that should be solved. PMID- 16044953 TI - [Clinical genetics for hereditary cancers: from the viewpoint of physicians working at a hospital specialized in cancer]. AB - In our daily practice, we provide clinical genetic consultation for patients at risk for hereditary cancers. The clinical characteristics of hereditary cancer syndromes in adults differ from those of hereditary diseases in children, although both involve genetic disease. One major difference is the difficulty in diagnosing hereditary cancers. Genetic testing has enabled us to diagnose HNPCC and familial breast and ovarian cancers. Another difference is the possibility to improve the outcomes of hereditary cancer syndromes by medical intervention. Intentional surveillance thus plays a key role in the management of hereditary cancer syndromes. These features make genetic counseling essential for hereditary cancer syndromes, including genetic testing and lifelong disease management. Networking among genetic disease specialists is particularly necessary. Clinical geneticists should be responsible for not only genetic disease but also for genomic information about cancer, with the ultimate goal of providing "order made" medical consultation and services. Another important goal is the establishment of systems for the comprehensive care of patients and for the career development of specialists to provide regional-based care for persons who have or are at risk for hereditary cancers, including future generations. PMID- 16044954 TI - [Necessity of registrar in medical institution]. AB - In our hospital, we are routinely investigating and recording patients with familial thyroid disorders. We found that about 30% of patients with thyroid disease had some relatives with the same disease as a result of the investigation of more than 13,000 patients. Therefore, a registrar is working for research of familial thyroid disease as a specialist in our hospital. The study was conducted on 258 patients with familial nonmedullary thyroid carcinoma (FNMTC). In cooperation with the registrar, the clinical characteristics of FNMTC emerged. Moreover, 15 patients with nonmedullary thyroid carcinomas were newly found among 149 asymptomatic FNMTC relatives (10%) and underwent operation. In this paper, we report the role of the registrar and emphasize the necessity of a registrar. PMID- 16044955 TI - [Gene testing of hereditary cancer on comprehensive gene medical examination support system]. AB - It has been estimated that genetic factors or a combination of genetic and environmental factors play a role in the development of 10-15% of all cancers. A genetic cause of hereditary cancer has been identified in more than 40 diseases till now. For preventing this cancer, gene testing is essential because it has no definite clinical marker as in hereditary non-polyposis colorectal cancer: HNPCC. Much more experience must be accumulated in this testing at the clinical base in order to increase specificity and sensitivity while safeguarding ethical, legal and social issues (ELSI). Recently, the Personal Information Protection Law was enforced. Gene inspection involving hereditary cancer should be carried out under a comprehensive gene medical examination organization. It is important for the family doctor, medical specialist, and gene inspection person in charge to cooperate closely with one another, and this will be a subject of future study. PMID- 16044956 TI - [The state of the art of hereditary cancer studies]. AB - During the last two decades, many genes responsible for hereditary cancer syndromes have been isolated. Based on the accumulating genetic information, genetic testing for both patients and the relatives is carried out in hospitals and clinics, and the clinical significance has been investigated. In addition to the genetic analyses of known genes and the functional analyses of the gene products, the recent research trends in hereditary cancer studies tend to move into the second era of research strategies including the isolation of novel genes responsible for remaining hereditary cancers, associated studies for finding cancer-susceptibility variants and the comprehensive analyses of expression profiles in tumors. Such new strategies are not only important to elucidate the pathophysiological mechanism of each hereditary cancer but may provide a potential application of tailor-made cancer therapy depending on the mutation status because many of the gene products seem to participate in the chemosensitivity of cancer cells. Furthermore, the research efforts have been expected to develop novel strategies for cancer prevention, diagnostics and therapeutics. In this paper, we have outlined the state of the art of studies on hereditary cancer syndrome, focusing on familial breast cancer and hereditary non polyposis colorectal cancer. PMID- 16044957 TI - [Genetic counseling]. AB - The emergence of molecular genetics into a routine medical service is demanding a paradigm shift in medical practice. An adequate reformation of its discipline and technology is required in every field of medical service including nursing. In 2000, the JFCR hospital founded a Familial Cancer Center to provide genetic counseling and genetic testing for cancer patents. Based upon our experiences with 250 families having various cancers, we have attempted to extract issues to be addressed in further detail. PMID- 16044958 TI - [Prevention of carcinogenesis in familial tumors]. AB - This report reviews the state of the art of cancer prevention of familial colorectal cancer (familial adenomatous polyposis, hereditary non-polyposis colorectal cancer). A large number of clinical trials have been performed using sulindac, a non-steroidal anti-inflammatory drug (NSAID). Sulindac reduces the size and number of large bowel polyps. However, as yet, it cannot be used for this indication in the clinical setting, because of the frequent occurrence of serious gastrointestinal side effects, and there are a number of cases in which aggressive tumors developed despite a reduction in the size of polyps. Studies on COX-2 selective inhibitors, with minimal side effects on the digestive tract, are showing promising results. In addition to NSAIDs, clinical trials have been performed using vitamins and dietary components. These show minimal side effects, but their efficacy is still insufficient for clinical use, and further studies are anticipated. PMID- 16044959 TI - [A Phase I study of docetaxel and cisplatin for advanced squamous cell carcinoma of the head and neck]. AB - A Phase I study of docetaxel (DOC) and cisplatin (CDDP) combination therapy was conducted as second-line treatment for advanced squamous cell carcinoma of the head and neck in order to determine the maximum tolerated dose (MTD), the dose limiting toxicity (DLT) of DOC, and the recommended dose (RD) for this combination therapy. Twenty patients with recurrence for whom failed first-line chemotherapy with CDDP and 5-FU proved in-effective (17 male, 3 female; age range 38-74 years; performance status 0=7, 1 =8, 2=5) were included in this study. DOC at dose level I (40 mg/m2), level II (50 mg/m2), level III (60 mg/m2) and level IV (70 mg/m2) was used, followed by CDDP administration at a fixed dose of 80 mg/m2. Originally, chemotherapy was repeated every 3 weeks. In level I, grade 4 hypokalemia occurred in one patient. No DLT occurred at level II. At level III, one patient experienced grade 4 vomiting. At level IV, grade 2 creatinine clearance decrease occurred in a total of two patients. The maximum tolerated dose in this combination therapy was DOC 70 mg/m2, and CDDP 80 mg/m2. The recommended dose for this combination therapy is DOC 60 mg/m2, and CDDP 80 mg/ m2. A multicenter cooperative phase II study in this RD is recommended. PMID- 16044960 TI - [Late phase II clinical study of KW-2307 in advanced/recurrent breast cancer patients (II)]. AB - A late phase II clinical study (II) of a novel vinca alkaloid derivative KW-2307 (vinorelbine ditartrate) in advanced/recurrent breast cancer patients was performed at 22 institutions throughout Japan. An intravenous dose of KW-2307, 20 mg/m2, was administered once a week. Of the 60 patients enrolled in the study, 58 were eligible and 56 were evaluable. The response rate was 33.9% (19/56; 95% confidence interval: 21.8 to 47.8%) with one CR and 18 PRs. The response rate was as high as 37.0% (17/46; 95% confidence interval: 23.2 to 52.5%) when KW-2307 was used as a first-line chemotherapy for advanced/recurrent disease. The most common adverse event was myelosuppression including leukopenia in 96.4% (54/56) and neutropenia in 94.3% (50/53). Other events observed were increased GOT in 51.8% (29/56), increased GPT in 55.4% (31/56), LDH increased in 50.0% (27/54), serum total protein decrease in 39.3% (22/56), anorexia in 41.1% (23/56), nausea and vomiting in 66.1% (37/56), constipation in 30.4% (17/56), alopecia in 33.9% (19/56) and general fatigue in 46.4% (26/56). None of them were serious. This study demonstrated that KW-2307 was an effective and safe treatment for advanced/recurrent breast cancer patients. PMID- 16044961 TI - [Retrospective analysis on efficacy and toxicity of 5-fluorouracil (5-FU) and l leucovorin (l-LV) in advanced or recurrent colorectal cancer]. AB - The aim of this study was to evaluate the efficacy and toxicity of 5-fluorouracil (5-FU) and l-leucovorin (I-LV) in 50 patients with advanced or recurrent colorectal cancer in our institute. The dose of 5-FU was 600 mg/m2 and the dose of l-LV was 250 mg/m2. Objective response were 36.8% of patients who had administration of full-dose and 14.8% of patients who had the administration of reduced dose or prolonged interval. No significant difference was observed in clinical benefit rates between patients administrated in full-dose and patients in reduced dose or prolonged interval. Median survival time (MST) of patients in reduced dose or prolonged interval is longer than patients in full-dose. These data suggest that 5-FU/l-LV can be given in the outpatient and yields improved prognosis and minimal adverse reactions even in patients in reduced dose or prolonged interval. PMID- 16044962 TI - [Multicenter comparative study of the recurrence-inhibitory effect of oral fluoropyrimidine drugs in patients with colorectal cancer following curative resection]. AB - HCFU and UFT were reported effective in adjuvant chemotherapy for colorectal cancer. This investigation was planned as a randomized study to compare the usefulness of combination therapies with mitomycin C (MMC)+HCFU and MMC+UFT as postoperative adjuvant chemotherapy in patients with colorectal cancer following curative resection, in terms of survival rate, recurrence rate, and adverse drug reactions. A total of 501 patients consisting of 252 patients with stage III/IV colon cancer (Colorectal Cancer Handling Rules, 4th Ed.) for which macroscopic curative resection was possible and 249 patients with stage II/III/IV rectal cancer (ibid, 4th Ed.) were registered from 40 participating institutions. The patients were randomly allocated to two groups with colon cancer and rectal cancer employed as stratification factors. Beginning on Day 14 after surgery, HCFU at 300 mg/day was administered to one group and UFT at 300 mg/day or 400 mg/day to another group, both orally and daily for one year. MMC 6 mg/m2 was administered intravenously to both groups on the day of surgery and the day following. Among the 501 patients, 496 patients (99%) were eligible. The 5-year survival rates were 77.1% for the MMC+ HCFU group and 79.2% for the MMC+UFT group, with the 5-year recurrence-free survival rates were 76.1% and 72.9%, respectively, neither showing a significant difference between the groups. Adverse drug reactions appeared in 23% of patients in the MMC+HCFU group and in 19% in the MMC+UFT group, with no serious reactions. One year after surgery the administration completion rates were good, at 82% for the MMC+HCFU group and 83% for the MMC+UFT group. No clear difference in effectiveness was noted between MMC+HCFU therapy and MMC+UFT therapy as postoperative adjuvant chemotherapy for colorectal cancer. The administration completion rates were good, and no serious adverse drug reactions were observed for either therapy. It was thus considered that both therapies could be administered safely, and both were useful as postoperative adjuvant chemotherapies for colorectal cancer. It is considered necessary to compare them with standard therapies in Western countries in the future. PMID- 16044963 TI - [Second-line chemotherapy with bi-weekly CPT-11 and cisplatin for recurrent colorectal cancer resistant to 5-FU based chemotherapy]. AB - Second-line chemotherapy with bi-weekly CPT-11 and cisplatin (CDDP) was given to 19 patients with recurrent colorectal cancer resistant to 5-FU based chemotherapy. The 19 patients consisted of 18 men and 1 woman with a mean age of 61.3 years. Nine patients had liver metastasis, 4 had lung metastasis, 2 had local recurrence, 2 had both local recurrence and lung metastasis, respectively, 1 had local metastasis and lymph node metastasis, and 1 had bone metastasis. CPT 11 (80 mg/m2) and CDDP (30 mg/m2) were administered bi-weekly. The objective overall response rate was 15.8%. The time to progression was 146 days, and the median survival time was 477 days. Grade 3 leucopenia and nausea occurred in 1 patient (5.3%). CPT-11 and CDDP treatment should be considered as second-line chemotherapy for colorectal cancer resistant to 5-FU based chemotherapy. PMID- 16044964 TI - [Cut-off level of docetaxel, paclitaxel and gemcitabine in histoculture drug response assay for non-small cell lung cancer]. AB - Cut-off levels of docetaxel (DOC), paclitaxel (PAC), and gemcitabine (GEM) in histoculture drug response assay are determined by data acquisition of non-small cell lung cancer. Inhibition rates were 47.5 +/- 22.2% in DOC (n=181), 66.6 +/- 25.1% in PAC (n=57), and 25.4 +/- 18.4% in GEM (n=63), respectively. Cut-off levels were determined as 50% in DOC, 60% in PAC, and 30% in GEM. The positive rates such as 47.5% in DOC, 68.4% in PAC, and 33.3% in GEM were obtained. PMID- 16044965 TI - [Establishment of enzyme-linked immunosorbent assay for quantification of orotate phosphoribosyltransferase in gastric carcinoma]. AB - A number of enzymes have been shown to be involved in the process of activation and/or degradation of 5-fluorouracil, and they are potential candidates for predicting factors of chemosensitivity to 5-fluorouracil. Among them, orotate phosphoribosyltransferase (OPRT EC 2.4.2.10) is a key enzyme related to the first step activation process of 5-fluorouracil and therefore it has been shown to be an important enzyme for the prediction of sensitivity to 5-fluorouracil and its related derivatives. We developed a new enzyme-linked immunosorbent assay (ELISA) system to accurately assess intratumoral activity of orotate phosphoribosyltransferase. A new sandwich ELISA system was established using anti OPRT polyclonal antibodies obtained from the rabbit immunized with the recombinant human peptides of the OPRT molecule. OPRT levels were measured in 8 human cancer xenografts transplanted in nude mice and 58 gastric cancer tissues using both a newly established ELISA and a conventional enzyme assay using radiolabeled 5-fluorouracil as a substrate. OPRT levels in 8 human cancer xenografts measured by this ELISA were significantly correlated with the OPRT enzyme activities (r2=0.782). Furthermore, OPRT activities measured in 58 gastric cancer tissues by enzyme assay were significantly correlated with those measured by the newly-established ELISA (r2=0.664). The ELISA system developed for the measurement of OPRT required a minimal amount of carcinoma tissue samples, which could be an easy-of-use assay system to predict sensitivity to 5-fluorouracil in gastric carcinoma. These results suggest that this newly-developed sandwich ELISA system for the quantification of OPRT is technically simple, feasible, and may be a useful tool to predict sensitivity to fluoropyrimidine-based anticancer chemotherapy in patients with gastric carcinoma and other cancers. PMID- 16044966 TI - [Pathologic complete response of thoracic esophageal cancer developing after gastrectomy to neoadjuvant low-dose nedaplatin (CDGP), 5-fluorouracil and radiotherapy]. AB - We treated a 69-year-old man who had developed esophageal cancer following gastrectomy. Pathologic complete response (pCR) was obtained by neoadjvant chemoradiotherapy using low-dose nedaplatin (CDGP) and 5-fluorouracil. The cancer located in the middle of the thoracic esophagus, had invaded the trachea and metastasized to cervical lymph nodes according to computed tomography. Preoperative chemoradiotherapy combining a low-dose of CDGP with 5-FU was administered together with radiotherapy. Adverse effects included grade 2 stomatitis and leukocytopenia. The esophageal cancer was found by endoscopy to have diminished significantly after completion of neoadjuvant therapy, An endoscopic biopsy specimen was found to contain no malignant cells. The tumor also was smaller by CT, where cervical lymph nodes no longer showed involvement. Partial response was diagnosed based on imaging, and radical resection of the esophageal cancer was performed via right thoracotomy and laparotomy. Operative staging findings indicated Ch x R-T 3 N 0 M 0, Stage II R 0 D 2 Cur A. Pedicled jejunum was used to reconstruct the esophagus through a mediastinal route. Pathologic examination of resected specimens disclosed no viable cancer cells in the esophagus or metastasis to dissected lymph nodes. Neoadjuvant chemoradiotherapy using low-dose CDGP/5-FU is an effective treatment for esophageal cancer. PMID- 16044967 TI - [Chemotherapy for an esophageal cancer patient undergoing continuous ambulatory peritoneal dialysis for chronic renal failure and measurement of plasma concentration of the drug]. AB - We administered chemoradiotherapy consisting of 5-fluorouracil (5-FU) combined with cisplatin, radiation and sequential chemotherapy using nedaplatin for an esophageal cancer patient undergoing continuous ambulatory peritoneal dialysis (CAPD) for chronic renal failure. There are no reports in the literature of chemotherapy for a patient maintained on CAPD. The dosage of each drug was based on that for a patient undergoing hemodialysis, and the plasma concentration of each drug was examined. Chemotherapy consisted of 3-10 mg/body of cisplatin with 60-min infusion and 450 mg/body of 5-FU with continuous infusion over 5 days. There were no side effects and no increase in the concentration of either of the drugs. Subsequently, 50 mg/body of nedaplatin, which is half the dose for a patient with normal renal function, was administered. The area under the blood concentration time curve (AUC) of nedaplatin was 15.85 microg/ml, which was slightly low compared with that after infusion of 80 mg/m2 in patients with normal renal function. Grade 3 leukopenia and thrombocytopenia occurred. As a final result, a partial response was obtained, and the patient was able to eat solid food after treatment. Although chemotherapy consisting of low-doses of cisplatin and 5-FU and a half dose of nedaplatin was administered safely, further study is needed to determine the suitable regimens for a patient maintained on CAPD. PMID- 16044968 TI - [TS-1 therapy via intestinal catheter used for tube feeding in a patient with gastric cancer after total gastrectomy]. AB - The patient was a 76-year-old man having gastric cancer with peritoneal dissemination. He underwent total gastrectomy for resection of the primary tumor and improvement of the oral intake. He developed ileus and peritonitis after the surgery, which necessitated two additional operations. An intestinal stoma was thus therefore created using a catheter for tube feeding, and tube feeding was initiated after the surgery. Nutrients, as well as TS-1 (taken out of the capsule; 80 mg/day) were administered via the catheter for tube feeding. This therapy was followed by a reduction in tumor marker levels and improvement of the patient's performance status (PS), after which the patient could be discharged. He stayed at home for about 8 months, with a much-improved quality of life during this period. We concluded that the TS-1 therapy via the catheter used for alimentation was effective for the treatment of cancer in this patient. We report our experience with this case, in which tube feeding became necessary after total gastrectomy, but administration of TS-1 via the same catheter used for alimentation improved the patient's PS and made it possible for him to receive chemotherapy at home. PMID- 16044969 TI - [A case of remnant gastric cancer with multiple bone metastasis and peritoneal dissemination; efficacy of combination therapy of docetaxel and TS-1]. AB - A 69-year-old female underwent radical surgery for advanced gastric cancer (Stage IIIA) 7 years ago. She was diagnosed as remnant gastric cancer with multiple bone metastasis and peritoneal dissemination. Treatment with docetaxel and TS-1 was started with the following regimen: daily oral administration of 100 mg/body TS-1 for 14 days, followed by a 7 day rest and infusion of 40 mg/m2 docetaxel on day 1. Two months after the initial administration of docetaxel/TS-1, the sites of the remnant gastric cancer and bone metastasis were reduced in size, and the ALP returned to almost the normal level. The site of peritoneal dissemination had disappeared. Currently (nine months after diagnosis), she is undergoing therapy with TS-1. The combination of docetaxel and TS-1 can be a new tool for the management of gastric cancer with bone metastasis. PMID- 16044970 TI - [A case of gastric cancer with peritoneal recurrence which developed during adjuvant administration of TS-1 showing complete response by weekly docetaxel regimen]. AB - A 62-year old man had undergone total gastrectomy for Borrmann type 4 gastric cancer. No peritoneal dissemination was observed at the laparotomy. Pathological examination revealed that the tumor involved the subserosal layer, and that the lymph node metastasis extended to the left gastric nodes. Vascular and lymphatic involvement was also observed. One hundred mg/body of TS-1, an oral 5 fluorouracil (5-FU) anticancer agent, which consisted of tegafur (a prodrug of 5 FU), and two modulators (gimeracil and oteracil potassium) was given from the 16th post-operative day. A course of TS-1 consisted of consecutive administration for 4 weeks followed by 2 weeks rest. The patient complained of abdominal fullness after administration of the second course of TS-1. Computed tomography (CT) revealed massive ascites. The serum carcinoembryonic antigen (CEA) titer was elevated to 13.5 ng/ml. From these findings, the occurrence of peritoneal dissemination was suspected. Weekly docetaxel of 30 mg/m2 (40 mg/body) was given for 3 weeks followed by a week cessation. At the start of the 6th course, the serum CEA was normalized, and CT scan detected the disappearance of ascites without any new lesion. Administration of docetaxel was continued until the 10th course then stopped without relapse of the disease. No dose reduction or postponement of administration were required. The patient has survived without disease one year after cessation of the treatment. Weekly docetaxel is a safe and effective regimen for gastric cancer worth using for a second-line therapy after failure of the 5-FU-based regimen. PMID- 16044971 TI - [Combined chemotherapy with oral leucovorin (LV) + tegafur/uracil (UFT) and hepatic arterial infusion (HAI) therapy for liver metastasis of colorectal cancer]. AB - The liver is the most frequent metastatic site from colorectal cancer, and the control of liver metastasis is an important issue in the treatment of progressive colorectal cancer. Hepatic arterial infusion (HAI) therapy can achieve a high drug concentration in the liver and relatively low level in the systemic circulation because of the first pass effect of the drug metabolism. With the high response rate, several reports have failed to show a significant survival benefit of HAI monotherapy, partially due to its inability to control extrahepatic metastasis. In this report, we used oral tegafur/uracil (UFT) and Leucovorin (LV) combined with HAI of 5-FU for four patients with liver metastasis of colorectal carcinoma. One of two patients with unresectable multiple hepatic metastases could undergo resectional surgery after 5 courses of this therapy. Two other cases in an adjuvant setting have been surviving free of tumors. In this series, adverse effects of this therapy were acceptable, including one case of grade 3 thrombocytopenia. The benefit of this combined therapy for survival in a case of liver metastasis from CRC remains to be evaluated. We are planning phase I and II clinical studies to evaluate the efficiency and feasibility of this combination therapy. PMID- 16044972 TI - [A case of cerebral infarction caused by disseminated intravascular coagulation during hepatic arterial infusion chemotherapy]. AB - A 73-year-old woman with liver metastasis underwent implantation of an infusion catheter-port system for hepatic arterial infusion chemotherapy. She developed multiple infarctions caused by disseminated intravascular coagulation (DIC) due to liver metastases. The hypercoagulability syndrome associated with cancer (known as Trousseau's syndrome) is considered a cause of cerebral infarction. Among the complications of the implantation of an infusion catheter-port system, Trousseau's syndrome may be one of the causes. PMID- 16044973 TI - [Combination chemotherapy with oral TS-1 and low-dose CPT-11 by hepatic arterial infusion for multiple hepatic metastases from colon cancer--a case report]. AB - Combined chemotherapy consisting of oral TS-1 and low-dose CPT-11 by hepatic arterial infusion is suggested to be a new effective treatment for multiple liver metastases from colorectal cancer. A 53-year-old man was diagnosed with multiple hepatic metastases from advanced colon cancer (Stage IV). The patient underwent partial resection of the colon and catheter insertion into hepatic artery for arterial infusion in November 2003. He was treated with postoperative combination chemotherapy consisting of UFT and low-dose CPT-11. UFT was administered orally at 400 mg/body/day everyday and CPT-11 was injected at 40 mg/body/week for 6 weeks, followed by a 2 weeks rest interval as 1 cycle. In spite of the reduction of metastatic liver tumors after 2 cycles of the chemotherapy, a metastatic pleural tumor appeared. Therefore, we judged the effect of the chemotherapy to be a progressive disease and changed UFT in the regimen to TS-1. TS-1 was administered orally at 80 mg/body/day under a 2-weeks-on and 1-week-off regimen for 3 times. CPT-11 was injected at 40 mg/body/week for 6 weeks, followed by a 3 weeks rest interval as 1 cycle. A stable disease was maintained for 3 months. Outpatient care was possible because no severe events were observed. Tumors showed a reduction rate of 37.4% after the combination therapy. The patient survived for 285 days after the operation. PMID- 16044974 TI - [Long-term survival in a case of large bowel cancer--efficacy of CPT-11-based chemotherapy]. AB - A 77-year-old woman underwent an ileocecal resection and a partial resection of the small intestine for cecal cancer. However, ileus caused by a recurrence in the small intestine was detected two years and four months postoperatively, so an ileal resection was performed. Furthermore, metastases to the lungs, lymph nodes, and peritoneum were detected, and CPT-11-based chemotherapy was administered. The patient was initially treated by combination therapy with a small dose of CPT-11 and CDDP. The combination drugs were changed to MMC, 5'-DFUR, etc. while the appearance of adverse reactions was monitored. Three years of continuous treatment served to prevent the proliferation of tumors. At present, TS-1 chemotherapy is ongoing. The results suggest that CPT-11-based chemotherapy can be continued by changing the combination of concomitant drugs while monitoring adverse reactions. It thus may be an effective regimen for advanced and recurrent large bowel cancer. PMID- 16044975 TI - [A case report of bowel obstruction due to peritonitis carcinomatosa using combined administration with CPT-11 and CDDP]. AB - We report here a case of recurrent gastric cancer that responded to third-line chemotherapy with CPT-11 and CDDP. The patient was a 61-year-old man with recurrent gastric cancer, who had administered TS-1 for first-line chemotherapy and paclitaxel for second-line chemotherapy. After such therapy, bowel obstruction was revealed due to peritoneal dissemination. The patient underwent third-line chemotherapy with CPT-11 and CDDP after drainage of gastrointestinal juice by nasogastric tube. The treatment schedule for CPT-11 and CDDP therapy consisted of CPT-11 60 mg/m2 div at day 1, day 15 and CDDP 60 mg/m2 div at day 1. It was repeated every 4 weeks. After first administration, the bowel obstruction was improved, so the treatment was continued for 8 months on an outpatient basis. These findings imply that this treatment can be a useful second-line or third line chemotherapy for unresectable advanced or recurrent gastric cancer. PMID- 16044976 TI - [Adjuvant chemotherapy of premenopausal breast cancer with LH-RH analogue for ovarian protection--a case report]. AB - We report a case in which the LH-RH analogue, goserelin acetate was administered to a 26-year-old female patient diagnosed with premenopausal breast cancer and concurrently receiving anthracycline-based adjuvant chemotherapy for ovarian protection. After radical operation, pathological diagnosis showed that adjuvant chemotherapy was indicated. As she hoped for childbirth, at first goserelin was injected twice and then adjuvant chemotherapy was undergone concurrently with goserelin acetate for ovarian protection. The adjuvant chemotherapy consisted of 4 cycles of every four week intravenous dripinjection of adriamycin 50 mg/m2 and cyclophosphamide 500 mg/m2. The chemotherapy and goserelin acetate were completed at the same time. Menstruation recovered about two months after the finish of adjuvant therapy and was well-regulated after recovery. It is suggested that goserelin combined adjuvant chemotherapy for premenopausal breast cancer may be useful for ovarian protection. PMID- 16044977 TI - America's best hospitals. PMID- 16044978 TI - Organizational barriers to physician participation in cancer clinical trials. AB - OBJECTIVE: To assess barriers to physician participation in cancer clinical trials among oncologists, oncology leaders, and health plan leaders. STUDY DESIGN: Mail survey of 221 oncologists combined with semistructured telephone interviews with oncology and plan leaders at 10 integrated healthcare systems. METHODS: The survey instrument examined physicians' involvement in clinical trials; their perception of the value of trials to them, their patients, and their organization; and the presence of infrastructure support for trials and associated resource constraints. The interviews investigated similar issues from the leaders' perspective. We used linear regression to model trial enrollment and standard qualitative techniques to analyze the interviews. RESULTS: Oncologists estimated they enrolled 7% of patients in trials. They expressed extremely favorable attitudes toward trials as a source of high-quality patient care and a benefit to themselves professionally. While positive attitudes toward trials were common, and were significant bivariate predictors of enrollment, organizational factors were the predominant predictors in multivariate analysis. The best combination of factors independently predicting enrollment related to organizational support for trials, subspecialty of the oncologist, and limitations of trial eligibility requirements. CONCLUSIONS: To increase trial participation, there is a critical need for infrastructure to support trials, especially additional support staff and research nurses. In addition, there is a need for better intra-organizational communication and consideration of the impact of trial design on internal health plan resources. This research supports the need to continue a national dialogue about the broadly defined benefits and costs of clinical trials to patients, physicians, and health plans. PMID- 16044979 TI - Association between physician compensation methods and delivery of guideline concordant STD care: is there a link? AB - OBJECTIVE: To examine the association between primary care physician (PCP) reimbursement and delivery of sexually transmitted disease (STD) services. STUDY DESIGN: Cross-sectional sample of PCPs contracted with Medicaid managed care organizations in 2002 in 8 California counties with the highest rates of Medicaid enrollment and chlamydia cases. METHODS: The association between physician reimbursement methods and physician practices in delivery of STD services was examined in multiple logistic regression models, controlling for a number of potential confounders. RESULTS: Evidence of an association between reimbursement based on management of utilization and the PCP practice of providing chlamydia drugs for the partner's treatment was most apparent. In adjusted analyses, physicians reimbursed with capitation and a financial incentive for management of utilization (odds ratio [OR] = 1.63) or salary and a financial incentive for management of utilization (OR = 2.63) were more likely than those reimbursed under other methods to prescribe chlamydia drugs for the partner. However, PCPs least often reported they annually screened females aged 15-19 years for chlamydia (OR = 0.63) if reimbursed under salary and a financial incentive for productivity, or screened females aged 20-25 years (OR = 0.43) if reimbursed under salary and a financial incentive for financial performance. CONCLUSION: Some physician reimbursement methods may influence care delivery, but reimbursement is not consistently associated with how physicians deliver STD care. Interventions to encourage physicians to consistently provide guideline concordant care despite conflicting financial incentives can maintain quality of care. In addition, incentives that may improve guideline-concordant care should be strengthened. PMID- 16044980 TI - Implementation of mandatory Medicaid managed care in Missouri: impacts for pregnant women. AB - OBJECTIVES: To assess the impact of mandatory Medicaid managed care in Missouri on prenatal care, maternal behavior, and low birth weight among pregnant women enrolled in Medicaid. STUDY DESIGN: Pre-post design using a comparison group with birth certificate and Medicaid enrollment data in 1995 and 2000. METHODS: Pregnant women delivering in 38 counties that implemented managed care in Medicaid were compared preimplementation and postimplementation with pregnant women delivering under Medicaid in 78 counties that remained fee-for-service (FFS) for separate samples of white (37,561) and black (13,640) non-Hispanic women. We calculated difference-in-difference estimates using linear probability regression models that controlled for maternal characteristics and time-invariant county differences. Analyses were stratified based on Medicaid enrollment before and after conception, managed care region, and marital status. RESULTS: Both managed care and FFS counties showed large improvements in prenatal care measures over time for both white and black women. Managed care was associated with a smaller percentage point increase relative to FFS counties in adequate care among whites of 1.9 and among blacks of 8.5, and a larger decrease in smoking of 4.8 and in Women, Infants, and Children Program enrollment of 2.3 among white women. No pattern across managed care regions was found with respect to timing of implementation. Smaller effects were evident among black and single women. CONCLUSIONS: Although women experienced significant improvements statewide in prenatal care under Medicaid, improvements were smaller for managed care counties. Managed care may have a positive impact on smoking cessation, but other policy changes may be needed to improve birth outcomes. PMID- 16044981 TI - Recruitment of Medicaid and dual-enrolled Medicare beneficiaries with diabetes mellitus into a randomized controlled trial. AB - OBJECTIVE: To document the recruitment of Medicaid and dual-enrolled Medicare beneficiaries with diabetes mellitus into a randomized clinical trial. STUDY DESIGN: Randomized controlled trial with 1-year follow-up. METHODS: A total of 2242 Medicaid or dual-enrolled Medicare beneficiaries with diabetes residing in King County, Washington, were recruited by direct mail for a clinical trial of diabetes self-care management. Washington State Medicaid program databases were used to identify the target population who received recruitment packets from the program director. Individuals who did not return a participation refusal letter were telephoned to determine study eligibility. Subjects were screened during a study visit, and written informed consent was obtained. Enrolled subjects were randomized to a self-care intervention group or a usual care group. RESULTS: Of 2242 recruitment packets sent, we were unable to contact 40% of the target population, despite the fact that packets were sent to the same mailing addresses used for monthly Medicaid check distributions. The primary recruitment challenges were missing telephone contact information and a lack of interpreters speaking needed dialects. Of the 146 subjects enrolled, 71% were nonwhite, 28% were non English speaking, 69% were women, and the mean age was 59.8 years. CONCLUSIONS: Research in Medicaid and Medicare populations is possible but requires additional time, energy, and resources. The finding that 40% of the Medicaid population could not be contacted suggests that Medicaid may want to revisit their contact information procedures to facilitate case management and other programs. PMID- 16044982 TI - Estimating medication persistency using administrative claims data. AB - OBJECTIVES: To review the definitions and methods for measuring medication persistency, and to propose a uniform definition of and calculation for persistency using pharmacy claims data. STUDY DESIGN: Literature review. METHODS: A MEDLINE search (1966 to present) was performed to identify articles detailing a definition or method of persistency measurement based on automated pharmacy data. Articles were screened for relevance by title and abstract. References from identified articles were used to expand the search results. RESULTS: The concept behind medication persistency measurement is to capture the amount of time that an individual remains on chronic drug therapy. The methods to calculate medication persistency can be classified into 1 of 3 categories: (1) Persistency as a function of the medication possession ratio; (2) persistency as a function of medication availability at a fixed point in time; and (3) persistency as a function of the gaps between refills. CONCLUSIONS: The common goal of all persistency measures should be to reflect the continuity of medication usage and to capture the timeliness and the frequency of refilling. The measurement of persistency as a function of the gaps between refills provides the best assessment of refill compliance across a variety of medication and disease states and lends itself to the well-established measurements of survival analysis. PMID- 16044983 TI - Starvation on the web. PMID- 16044984 TI - A jury of their peers. PMID- 16044985 TI - The quiet power of the stoic. PMID- 16044986 TI - Labouring for motherbaby. PMID- 16044987 TI - Student midwives win landmark case. PMID- 16044988 TI - How women choose where to give birth. PMID- 16044989 TI - Does pethidine relieve pain? PMID- 16044990 TI - Home birth: safe as houses? PMID- 16044991 TI - Developing valuable assets. PMID- 16044992 TI - Getting parent education right. PMID- 16044993 TI - Is the Apgar a flexible friend? PMID- 16044994 TI - Don't mention the 'B' word! PMID- 16044995 TI - Prostaglandins for the prevention of PPH. PMID- 16044996 TI - Singing from the same hymn sheet? PMID- 16044998 TI - Counting your blessings. PMID- 16044999 TI - Therapeutic relationship with children and family. PMID- 16045000 TI - Asthma camp. PMID- 16045001 TI - Connecting for children's health. PMID- 16045002 TI - The roles and responsibilities of newly qualified children's nurses. AB - The roles and capabilities of newly qualified nurses have been studied in the past but little is known about whether newly qualified children's nurses undergo similar transitions. Six recently qualified children's nurses were interviewed in this small qualitative study to explore the meaning of being a staff nurse and identify factors that facilitated or inhibited transition from child branch student to registered nurse. A theoretical framework of role transition and situated learning was derived from the interview data. The findings indicated that becoming a staff nurse is a process leading to an identity that is actively and continuously being created PMID- 16045003 TI - Managing capacity and workload in children's services. PMID- 16045004 TI - The effects of eczema on children and their families: a review. PMID- 16045005 TI - Is restraint a form of abuse? PMID- 16045006 TI - [Effects of CD147 on the production of matrix metalloproteinase-9 by fibroblasts and the invasion of melanoma cells]. AB - OBJECTIVE: To investigate a possible role of CD147 in the production of matrix metalloproteinase-9 (MMP-9) by fibroblasts and the invasion of melanoma cells. METHODS: We cocultured CD147-expression melanoma cells with fibroblasts and examined the MMP-9 expression of fibroblasts by zymography and the invasion of melanoma cells by transwell invasion assay. RESULTS: MMP-9 expression was enhanced in conditioned media, while cocultured with melanoma cells in a dose dependent manner, and CD147 antibody inhibited the production of MMP-9 in the fibroblasts. When fibroblasts were cultured at the bottom of the lower compartment of transwell invasion model, the number of melanoma cells that invaded significantly increased. Addition of anti-CD147 antibody to the upper compartment transwell invasion model resulted in the significant inhibition of the melanoma cell invasion in reconstituted basement membrane. CONCLUSION: CD147 expressed in melanoma cells plays an important role in the melanoma cell invasion by stimulating the production of MMP-9 by fibroblasts. PMID- 16045007 TI - [Stable suppression of beta-catenin expression in prostate cancer cell line by retrovirus mediated RNAi]. AB - OBJECTIVE: To set up a prostate cancer cell line in which beta-catenin expression is stably suppressed and to investigate the role of Wnt/beta-catenin signaling pathway in prostate tumorgenesis. METHODS: We select 3 sites in the complete coden sequence region of beta-catenin gene as the RNAi targets, ligated the annealed double pre-DNA strands into the retroviral vectors pSUPER-retro and transfected them into the packaging cells PA317, and then collected supernatant with retrovirus to infect DU145. After selection by puromycin and culture expansion, the stable cell clones were attained. Expression of the 2 target genes of Wnt/beta-catenin signaling pathway cyclinD1 and c-myc, was detected in the beta-catenin RNAi cells by Western blot. The effect of suppressing beta-catenin by RNAi on cell proliferation was quantified by methylthiazoletetrazolium (MTT) assay. RESULTS: Western blotting and RT-PCR showed that the expression level of beta-catenin in the 2 stable cell clones apparently decreased. CyclinD1 and c-myc expression decreased in the beta-catenin RNAi cells. MTT showed that the cell number of beta-catenin expression suppression cell clones decreased significantly (P < 0. 05), suggesting the cell proliferation was prevented. CONCLUSION: The beta-catenin gene stable suppression cell line was successfully established. PMID- 16045008 TI - [Cytogenetic study of 1780 cases of spontaneous abortion]. AB - OBJECTIVE: To explore the relationship between chromosome anomaly and spontaneous abortion, and to provide useful information for genetic counseling and prenatal diagnosis in reproductive clinic. METHODS: A total of 1 780 patients who had a history of spontaneous abortion before 24 weeks of gestation were enrolled. The lymphocyte culture and harvest were performed according to standard methods. Karyotypes were analyzed by G-banding in all cases and C- banding in some cases in addition. RESULTS: Altogether 57 abnormal karyotypes were found and the overall incidence of chromosomal abnormalities was 3.20% (women 3.32%; men 2.12%). Among them 23 cases were the balanced translocation; 14 cases were the Robertsonian translocation, 3 cases were the complex chromosomal rearrangement, and the other 17 cases were the other abnormalities. In women with 1, 2, 3 or more spontaneous abortion, the incidence of chromosomal abnormalities was 1.7%, 2.3%, and 5.8%, respectively. CONCLUSION: Translocations are the major abnormal karyotpes associated with spontaneous abortions. The chance of finding chromosomal aberration increases with the number of abortions. Chromosomal abnormalities are more common in women with 3 or more spontaneous abortions. PMID- 16045009 TI - [Effects of sodium aescinate on bcl-2 and caspase-3 expression and apoptosis after focal cerebral ischemia reperfusion injury in rats]. AB - OBJECTIVE: To determine the effects of sodiun aescinate on Bcl-2 and Caspase-3 protein expression and neuronal apoptosis after focal cerebral ischemia reperfusion injury in rats. METHODS: One hundred male Wistar rats were subjected to middle cerebral artery occlusion (MCAO) and reperfusion. The rats were divided randomly into 4 groups: sham-operated group and MCAO and reperfusion model groups which were randomly divided into control group, saline group, and sodium aescinate group. The immunohistochemistry staining and microscope image were used to observe the dynamic Bcl-2 and Caspase-3 protein expression in the ischemic penumbra after the reperfusion. Terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) staining was performed for the detection of apoptosis. RESULTS: Bcl-2 protein expression in the sodium aescinate group was significantly higher than that of the saline group and control group (P < 0.05). While Caspase 3 protein expression in the sodium aescinate group was then compared with those of the saline group and control group, and showed the difference was significant (P < 0.05). Compared with the saline group and control group, the number of apoptotic cells in the sodium aescinate group was significantly reduced (P < 0.01). CONCLUSION: Sodium aescinate increases Bcl-2 protein expression and decreases Caspase-3 protein expression,through which it can protect the ischemia brain on reperfusion injury. PMID- 16045010 TI - [Effect of hepatocyte growth factor on oxygen-glucose deprived injury of astrocytes]. AB - OBJECTIVE: To explore the effect of hepatocyte growth factor (HGF) on oxygen glucose deprived injury and apoptosis of astrocytes. METHODS: The injury of primary cultured rat cerebral cortical astrocytes was induced by oxygen-glucose deprivation. Astrocytes were treated with HGF at various final concentrations of 20 - 100 ng/mL. The cell damage and viability were evaluated by the lactate dehydrogenase (LDH) released rate and the 3- (4,5-dimethylthazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) conversion method. Detection of apoptotic cells was determined by the flow cytometry, and the ultrastructure was observed by the transmission electron microscope. RESULTS: Oxygen-glucose deprivation increased the LDH release rate, decreased the cell viability and increased the number of apoptotic astrocytes. While exposed to HGF at the same condition, the LDH release rate decreased, the cell viability increased, and the percentage of apoptotic cells decreased (P <0.05). The maximum protective effect of HGF was observed at 60 ng/mL. CONCLUSION: HGF can protect cultured astrocytes from oxygen glucose deprived injury, and attenuate the apoptosis of astrocytes in a dose dependent manner. PMID- 16045011 TI - [Differentiation of mesenchymal stem cells into cardiomyocytes induced by cardiomyocytes]. AB - OBJECTIVE: To investigate the role of adult cardiomyocytes in the differentiation of mesenchymal stem cells (MSCs) into cardiomyocytes. METHODS: Rat MSCs were isolated by a Percoll's gradient solution and cultured in low-glucose Dulbecco' s modified Eagle' s medium (DMEM). After 2 passages, cell-surface antigen CD34, CD71 and CD90 for rat MSCs were determined by flow cytometry, and these MSCs were transfected with pEGFP-N3 by Lipofectamine2000. Then those MSCs labeled with GFP, were cultured in contacted, nocontacted and conditioned with adult rat myocardiocytes. Immunofluorescence staining against alpha-actin, desmin, and troponin-T were performed after 1 week. RESULTS: Immunofluorescence staining was positive against alpha-actin, desmin, and troponin-T on MSCs in contacted culture group. In contrast, no alpha-actin, desmin, and troponin-T expression on MSCs were observed in the noncontacted culture group and the conditioned culture group. CONCLUSION: Direct cell-to-cell contact between MSCs and adult cardiomyocytes may induce differentiation of MSCs into cardiomyocytes. PMID- 16045012 TI - [Prevalence and influence factors of hypertension among mechanic factory workers]. AB - OBJECTIVE: To determine the status and influence factors of hypertension on mechanic factory workers and to provide reference for further hypertension prevention and control. METHODS: A cross-sectional study on 1205 workers (exposed to different noise levels) in Hunan was carried out by using questionaire and measuring the blood pressure of the workers and the noise exposure level in the workplace. The prevalence and the influence factors of hypertension among mechanic factory workers were analysed. RESULTS: The hypertension prevalence was 12.1%. Logistic regression analysis showed the body weight index (BMI), age, and history of hypertension in parents and accumulative noise dose levels influenced the hypertension prevalence. CONCLUSION: Controlling the body weight, reducing alcohol consumption, decreasing the sound pressure level in workshops and advocating healthy diet may reduce the prevalence rate of hypertension among mechanic factory workers. PMID- 16045013 TI - [Effect of simvastatin on cardiac fibrosis in patients with essential hypertension]. AB - OBJECTIVE: To explore the effect of simvastatin on myocardiac fibrosis in patients with essential hypertension (EH). METHODS: Sixty EH patients were randomly assigned into 2 groups: Benazepril (10 mg/d) group (n = 28) and simvastatin (20 mg/d) + benazepril (10 mg/d) group. Procollagen type III aminoterminal peptide (PIIIP), and procollagen type IV aminoterminal peptide (PIVP) levels in serum as well as transforming growth factor beta 1 (TGFbeta1) level in plasma were measured by radioimmunoassay (RIA) before and 6 months after the treatment. Doppler ultrasound recordings were obtained from all patients before and 6 months after the treatment to determine several parameters related to the left ventricular anatomy and function. RESULTS: After 6 month of treatment, the mean blood pressure (MBP), PIIIP, PIVP, TGFbeta1, left ventricular mass index (LVMI), interventricular spectum dimension (IVSD), and left ventricular posterio wall dimension (LPWD) in the 2 groups were significantly lower than those before the treatment. TGFbeta1 decreased in the simvastatin and benazepril group compared with the benazepril group (P < 0.01). The ratio of early diastolic blood flow velocity of mitral valve (VE) and blood flow velocity of atrium systolic period (VA) in the 2 groups significantly increased after 6 months of treatment, and the ratio in the simvastatin and benazepril group was significantly higher than that in the enazepril group (P < 0.05). CONCLUSION: Angiotension converting enzyme inhibitor combined with simvastatin is helpful to reduce the myocardial fibrosis and to improve the left ventricular hypertrophy and diastolic function in EH patients. PMID- 16045014 TI - [Effects of astragalus polysaccharides-chitosan/polylactic acid composite material on biological behavior of canine bone marrow stromal cells cultured in vitro]. AB - OBJECTIVE: To observe the biological behavior of canine bone marrow stromal cells (BMSCs) cultured in vitro with the astragalus polysaccharides-chitosan/polylactic acid (AP-C/PLA) and with the chitosan/polylactic acid (C/PLA) and to find a suitable compound material for periodontal tissue engineering. METHODS: BMSCs (induced 14 days by 50 mg/L vitamine C, 10(-8) mol/L dexamethasone, 10 mmol/L beta-sodium glycerylphosphate) were cultured on AP-C/PLA or C/PLA for 5 days respectively. The BMSCs attachment and the morphology were observed with scanning electronic microscope and the combining rates were counted. Type I collagen synthesis was examined with immunohistochemistry staining and the content of osteocalin was determined with radio-immunological method. RESULTS: Combining rates, type I collagen synthesis, and the content of osteocalin of BMSCs on AP C/PLA were significantly higher than those on C/PLA. CONCLUSION: AP-C/PLA may promote the BMSC proliferation, differentiation and extracellular matrix synthesis, and it can be used as a good scaffold material for bone tissue engineering. PMID- 16045015 TI - [Differentiation-inducing effects of perphenazine on K562 leukemia cells]. AB - OBJECTIVE: To determine the differentiation-inducing effects of perphenazine on K562 leukemia cells. METHODS: Differentiation-Inducing effects of a phenothiazine perphenazine were evaluated by proliferation, morphology and function of K562 cells. We evaluated the effects of perphenazine on K562 cells proliferation by cellular enumeration in liquid culture assay, MTT assay and clony formation assay, the morphology by Wight-Gimesa staining, and the function by detecting CD71 through flow cytometry. RESULTS: Perphenazine enhanced the expression of CD71 on K562 cells and increased Hb content in K562 cells, while inhibited the proliferation of K562 cells. K562 cells showed differentiation morphology after the drug treatment. CONCLUSION: Perphenazine possessed differentiation-inducing effects on K562 cells. PMID- 16045016 TI - [Detection of glutathione S-transferase and lung resistance-related proteins in acute leukemia and its clinical significance]. AB - OBJECTIVE: To explore the relationship among intracellular glutathione S transferase activity (GST), the expression of lung resistance-related proteins (LRP) in acute leukemia, and its clinical effects. METHODS: The GST activity of bone marrow mononuclear cells and LRP expression in 57 acute leukemia patients were detected by the spectrophotometry assay and immuno-cytochemistry (SABC), respectively. RESULTS: The GST activity of bone marrow mononuclear cells in the acute leukemia group was significantly higher than that of the control group (P < 0.01). The GST activity of mononuclear cells in acute leukemia was positively correlated with the percentage of blast in the bone marrow (r = 0.30, P < 0.05). The GST activity of mononuclear cells in the untreated acute leukemia group was obviously higher than that of the complete remission group (P <0.01). The GST activity in the refractory or relapsed acute leukemia group was significantly higher than that of the complete remission group and untreated leukemia group (P <0.05). In post-chemotherapy 13 of 17 the LRP-positive patients were the non remission, 12 of the 20 LRP-negative patients were the complete remission. The curative rate of the LRP-positive group was the significantly lower than the LRP negative group (P < 0.05). The GST activities of non-remission patients in the LRP-positive and LRP-negative group obviously increased. CONCLUSION: The increase of GST activity in the bone marrow mononuclear cells is related to the clinical curative effects and the proliferation of blast in acute leukemia. Detection of LRP and GST activities in acute leukemia may have a reference value in judging the leukemia with drug resistance and estimating the prognosis. PMID- 16045017 TI - [Dynamic changes of MMP-2 activity in the posterior sclera of chicks with form deprivation myopia]. AB - OBJECTIVE: To investigate the effect of form-deprivation on level of gelatinase in the posterior sclera in chicks. METHODS: Fifty 1-day-old chicks were monocularly deprived to establish the animal model of form-deprivation myopia (FDM). According to the duration of form-deprivation the experimental chicks were divided randomly and equivalently into 5 groups, which were deprived for 3, 7, 14, 21 and 30 days respectively. Meanwhile the other eyes of the deprived chicks were used as self-control groups and chicks of the same days were chosen randomly as the normal control groups for each FDM group. At each form-deprivation point the changes of degree of diopters and axial length of chicks in each group were recorded. The levels of gelatinase in posterior sclera of the experimental eyes were measured by gelatin enzymography. RESULTS: Compared with the normal and self control groups, the levels of MMP-2 activity in FDM groups were much higher (P <0.01). With the increase of the time of monocular deprivation these changes became more significant and reached the top after 14 days' deprivation with an inter-group statistical difference (P <0.01). The dynamic changes of MMP-2 activity were the same as those of axial length and degree of diopters in each experimental groups. There was positive correlation between the MMP-2 activity and axial length (r = 0.989, P < 0.01). But there was a negative correlation between the MMP-2 activity and refractive degree. CONCLUSION: Increase of MMP-2 activity in the posterior sclera of chicks would be a direct key factor to trigger sclera ECM remodeling process in chick FDM. PMID- 16045018 TI - [Quality of life in the treatment assessment of postmenopausal osteoporosis]. AB - OBJECTIVE: To evaluate quality of life (QOL) in the treatment assessment of postmenopausal osteoporosis (PMOP) by comparing the QOL of three treatments: hormone replacement treatment, supplement of calcium and vitamin D and risedronate. METHODS: All patients with postmenopausal osteoporosis (PMOP) in this clinical trial had been allocated into 4 groups: placebo therapy group, vitamin D addition calcium therapy group, hormone replacement treatment (HRT) therapy group, and risedronate therapy group. We measured the bone mineral density (BMD) and quality of life sale (QOLS) of patients in three times, before the treatment, 3 months after the treatment,and 12 months after the treatment. The differences of the QOL and BMD at the 4 groups and at different time, the linear correlation of the change of QOL and the change of BMD, were both compared after the measurement. RESULTS: The total score of QOL and the score of disease domain and physical domain of HRT therapy group and risedronate therapy group were higher than the placebo therapy group (F = 17. 335, P <0.001), but vitamin D addition calcium therapy group was not different from that of the placebo therapy group. The score of other three domains had no statistically significant difference among the 4 groups. The score of disease domain of the patients of risedronate therapy group started to increase after 3 months of treatment, and continued to increase after 12 months of treatment. The score of disease domain of the patients of HRT therapy group started to increase only after 12 months of treatment. But the score of disease domain of the patients of vitamin D addition calcium therapy group did not increase after 12 months of treatment. The changes of the score of disease domain had the linar correlation with the changes of BMD, and the correlation coefficient was from 0.608 to 0.827. CONCLUSION: QOL may become one of the indexes of medical treatment outcome assessment system for PMOP. PMID- 16045019 TI - [Evaluation of tumor necrosis factor-alpha and tumor necrosis factor-beta in serum of patients with endometriosis]. AB - OBJECTIVE: To evaluate the levels of tumor necrosis factor-alpha (TNF-alpha) and tumor necrosis factor-beta (TNF-beta) before and after conservative laparoscopic surgery in patients with endometriosis. METHODS: The levels of TNF-alpha and TNF beta in the serum of both 82 patients with EMS and 68 controls were determined by ELISA. RESULTS: The levels of TNF-alpha and TNF-beta in the serum of patients with EMS were significantly higher than those of the controls (P < 0.01), and they increased with the clinical terms ( P < 0.05). After clearance of endometrosis foci with laparoscopic conservative surgery, the TNF-alpha levels decreased significantly in EMS III - IV, and TNF-beta levels decreased significantly in EMS I - IV. CONCLUSION: Measuring TNF-alpha and TNF-beta levels in the serum of patients with EMS may have important value in postoperative follow-up, surveillance and evaluation of the effectiveness of the surgery. PMID- 16045020 TI - [Effect of progesterone on the secretion of matrix metalloproteinase-2 and matrix metalloproteinase-9 in human ectopic endometrial stromal cells]. AB - OBJECTIVE: To determine the effect of progesterone on the secretion of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in ectopic endometrial stromal cells. METHODS: Ectopic endometrial stromal cells were obtained from 17 patients with endometriosis. Endometrial stromal cells were obtained from 12 patients with endometriosis and 14 cases of controls. Ectopic endometrial stromal cells of 15 cases were treated with progesterone. Culture supernatants of these stromal cells were analyzed for MMP-2 and MMP-9 by zymography. RESULTS: Endometriotic stromal cells released significantly higher levels of MMP-2 and MMP-9 than endometrial stromal cells from women with and without endometriosis. Progesterone at 10(-9) mol/L caused endometriotic stromal cells a significant reduction MMP-2 and MMP-9 levels. When progesterone concentration was increased from 10(-9) mol/L to 10(-7) mol/L, the release of MMP 9 was almost completely inhibited, wherease that of MMP-2 was not completely inhibited. CONCLUSION: Progesterone may inhibit the secretion of MMP-2 and MMP-9 in ectopic endometrial stromal cells, especially MMP-9. PMID- 16045021 TI - [Expressions of survivin and nm23 and the relationship among the expressions and the axillary lymph node metastasis in breast cancer]. AB - OBJECTIVE: To investigate the expressions of Survivin protein and nm23 protein and the relationship among the expressions and axillary lymph node metastasis in breast cancer. METHODS: The expression of Survivin and nm23 in 80 cases of breast cancer tissues were detected by immunohistochemistry SP method, and their correlation with axillary lymph node metastasis and 5-year disease free survival (DFS) were analysed. RESULTS: Survivin protein positive expression rate was 68.75% (55/80) in breast cancer tissues, which had positive correlation with the axillary lymph nodes metastasis but negative correlation with 5 years FS (P < 0.05); nm23 protein expression had negative correlation with the axillary lymph nodes metastasis but positive to 5 years FS (P < 0.05). Survivin and nm23 proteins expression had no obvious correlation with the breast cancer pathology type, patient age and clinical stage (P > 0.05). CONCLUSION: The anti-apoptosis effect of Survivin protein and the anti-metastasis effect of nm23 protein may be important in the occurrence and advancement of breast cancer, suggesting that it may be a new indicator of prognostic and judgement in breast cancer. PMID- 16045022 TI - [Sex hormones in female patients with systemic lupus erythematosus]. AB - OBJECTIVE: To explore the levels of sex hormones in female patients with systemic lupus erythematosus (SLE) and its role in the genesis of SLE. METHODS: The serum levels of estradiol, estriol, testosterone, progesterone, and prolactin were determined by electro-chemiluminescence immunoassay in 98 female patients with SLE and compared with those of 38 healthy women. RESULTS: The serum levels of estradiol, estriol, progesterone, and prolactin were significantly higher than those in the healthy women (P <0.05). The serum levels of estradiol, progesterone, and prolactin in patients with SLE in 25 to 34 year old group were higher than the other age groups and the control group (P < 0.05), and the serum levels of estradiol and prolactin in patients with active phase of SLE were significantly higher than those in patients with stable phase of SLE (P <0.05). CONCLUSION: The levels of sex hormones have a close corretation with the genesis and development of SLE. PMID- 16045023 TI - [Sensitivity of nanoparticlized cefazolin sodium to the bacteria in vitro]. AB - OBJECTIVE: To investigate whether the in vitro sensitivity of cefazolin sodium to the bacteria was altered after nanoparticlization. METHODS: The minimal inhibitory concentrations (MIC) of cefazolin sodium before and after nanoparticlization to S. aureaus and E. coli. were determined by microdilution. RESULTS: The MIC of nanoparticlized cefazolin sodium to S. aureaus and E. coli. had no significant change compared with that of non-nanoparticlized one. CONCLUSION: Nanoparticlization will not decrease the sensitivity of cefazolin sodium to the bacteria. PMID- 16045024 TI - [Effects of leukocyte depletion filter on canine kidney during cardiopulmonary bypass]. AB - OBJECTIVE: To explore the effect of a reforming leukocyte depletion filter (LDF 1) on the functional and pathologic changes of canine kidney during cardiopulmonary bypass (CPB). METHODS: Twelve Mongolian dogs were randomly allocated into a control group (no LDF-1, n = 6) or a leukocyte-depleted filter group (LDF-1 placed in venous line, n = 6). CPB of the dogs anestheitized with sodium pentobarbitone at 25 mg/kg was set up. After 10 min of CPB, aorta was clamped and St. Thomas cardioplegic solution at 20 mg/kg was immediately injected into the root of aorta. The aortic cross-clamp was released and CPB was closed at 70 min. Dogs were observed for 2 h after weaning from CPB. LDF-1 was opened at 2 min and stoped at 7 min during initially running CPB in the LDF-1 group. Circulating leukocytes, plasma L-selectin, and plasma IL-8 were respectively counted before CPB, at 10 minutes, 40 min, and 75 min during CPB, the end of CPB, and 2 h after CPB. The urine analysis and renal pathology, which were obtained before CPB and 2 h after CPB, were observed. RESULTS: The number of leukocytes significantly decreased by 55% - 68% in the LDF-1 group compared with the baseline during CPB. The value at 10 min of CPB in the LDF-1 group was lower than that in the control group (P < 0.05). Plasma levels of L-selectin and IL-8 obviously increased in the 2 groups compared with the baseline during CPB, but both levels at 2 h after CPB in the LDF-1 group were lower than those in the control group (P <0. 05). No statistic difference was found in plasma levels of urea and creatinine, but hematuria was observed in the 2 groups at 2 hours after CPB. The pathologic changes of kidney, which was mainly renal tubule swelling accompanied partly with vacuolar degeneration, were similar under the light microscope in the 2 groups at 2 h after CPB. Obvious glomerular damage was not found. CONCLUSION: LDF-1 can effectively decrease leukocyte counts and the inflammatory reaction, but it can not bring about excellent protective effect on kidney during CPB when used alone. Attention to should be paid the renal protection in the postoperative CPB. PMID- 16045025 TI - [Isobaric and hyperbaric local anesthetic used in spinal anesthesia]. AB - OBJECTIVE: To reveal the advantages and disadvantages of the application of isobaric and hyperbaric local anesthetic in spinal anesthesia so as to provide reference for clinical practice. METHODS: One hundred and sixty ASA patients (physical status I - II) undergoing lower abdominal surgery within 3 hours under spinal anesthesia (using CSEA technique via spinal needle in epidural needle) were allocated to 2 groups with 80 cases each. In lateral decubitus, patients randomly received a subarachnoid injection of 3.0 mL (15 mg) isobaric (Group I) or hyperbaric (Group H) bupivacaine and then turned supine. Hemodynamic changes and patients' responses were perioperatively observed. After subarachnoid injection, we recorded the time of onset and motor block, the peak sensory blocked level, the time of regression of 2 dermatomes, the time of the first administration of analgesics for a significant pain of the incision, the time of the regression of motor block to modified Bromage scale 2, and the time of recuperating the function of urination. RESULTS: Both isobaric and hyperbaric 0.5% bupivacaine solutions in a volume of 3.0 mL provided effective sensory and motor block for the operations. The time of onset and complete motor block were similar in the two groups. Compared with Group I, the time of peak sensory block in Group H was shorter, the peak sensory block level was higher (more maximal dermatomes of blocked sensory nerves), the time of regression of sensory and motor block were shorter, the time of recuperating the function of urination was longer, and the incidence of feeling sick, nausea, vomiting and hypotension was higher. CONCLUSION: Isobaric solution is superior to hyperbaric solution in spinal anesthesia. PMID- 16045026 TI - [Surgical treatment of partial atrioventricular septal defect]. AB - OBJECTIVE: To summarize the experience of surgical treatments of partial atrioventricular septal defect in 60 patients. METHODS: From April 1999 to April 2004, 60 patients of partial atrioventricular septal defect were operated. Fifty eight patients were performed with suture of the cleft of mitral valve and the other 2 were given mitral valve replacement; For closure of primum ASD, 53 patients with pericardial patches and 7 with Dacron patches. Coronary sinus was baffled to left atrium with kirklin procedure in 35 cases and baffled to right atrium with McGoon procedure in other 25 cases. Correct the accompanying cardiac deformity at the same time. RESULTS: The hospital mortality was 3.3% (2/60) due to low cardic output syndrome. The incidence rate of complete atrioventricular block was 8.00% (2/ 35) in the group with Kirklin procedure and 6.06% (2/25) in the group with McGoon procedure. There was no statistical significance between the 2 groups (P > 0.05). The follow-up was from 1 month to 5 years, and there was no late death. All cardiac function were improved except middle mitral regurgitation in 1 patient. CONCLUSION: Reasonable operative design, refined procedures, avoiding damage to conducting bundles and proper perioperative management are the key points in improving theraeutic effect. PMID- 16045027 TI - [Two-stage administration sedation method for gastroscopy in hypoxia predisposed patients]. AB - OBJECTIVE: To explore the safety and the efficacy of two-stage administration sedation method (with propofol plus midazolam) in the analgesia gastroscopy for hypoxia predisposed patients. METHODS: Sixty-four hypoxia predisposed patients (with snore or expectoration) were divided into two groups and received sedation respectively with two-stage administration method and routine continuum administration method before gastroscopy. We analyzed the alteration of SBP, DBP, SpO2, sedation level, the degree of pharyngeal malaise, recovery time, dose of propofol,the lethe degree of upset, and procedure of gastroscopy to evaluate the safety and the efficacy of the two methods in the analgesia gastroscopy for hypoxia predisposed patients. RESULTS: Compared with routine continuum administration sedation method, two-stage administration sedation method had little influence on SpO2, so it reduced the accident of respiratory arrest (P < 0.01). The gastroscopy could be achieved at the lower sedation level with two stage administration method (P <0.01), so that the recovery time was shortened (P < 0.01). The degree of pharyngeal malaise in the patients with two-stage administration sedation method was higher than that of patients with routine continuum administrationsedation method (P < 0.01), and the dose of propofol used for sedation decreased (P < 0.01). While the lethe degree of gastroscopy upset and alteration of SBP, DBP were rated similarly in both groups (P > 0.05), and the lethe degree of gastroscopy procedure in two-stage administration method group was significantly lower than that of routine continuum adminis- tration method group (P <0.01). CONCLUSION: The two-stage administration sedation method is a new effective and safe sedation method for gastroscopy in the hypoxia predisposed patients. PMID- 16045028 TI - [Relationship between angiogenesis and expression of HO-1 of scar]. AB - OBJECTIVE: To investigate the relationshion between the angiogenesis of different kinds of scar and expression of HO-1. METHODS: The expression of heme oxygenase-1 and vessel counted by CD34 of biopsies from different kinds of scars such as hypertrophic scar, keloid, surgical scar and normal skin of 24 cases was valued by immunochemical method, and the relationship was compared between them. RESULTS: The vessel count of hypertrophic scar, keloid was significantly abundant compared with surgical scar or normal skin (P < 0.01). While the expression of HO 1 of hypertrophic scar, keloid was obviously higher than that in surgical scar or normal skin (P < 0.01), decreased from hypertrophic scar, keloid, surgical scar to normal skin. There existed a positive correlation between vessel count and the expression of HO-1 (r = 0. 761, P < 0.01) as well as the number of fibroblastic cells (r = 0. 731, P < 0.01) in the study groups. CONCLUSION: HO-1 might play a important role in the angiogenesis of scar formation. The cause of these changes may be local. Over angiogenesis is one symbol of pathological scar. PMID- 16045029 TI - [Microvessel counts and the expressions of chemotactic factors in the pathological scar tissues]. AB - OBJECTIVE: To explore the microvessel counts and the expressions of interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1), and macrophage inflammatory protein-1 ( MIP-1) alpha mRNA in the pathological scar tissues. METHODS: Immunohistochemical method of avidin-biotin complex was used for microvessel counts on the routinely formalin-fixed and paraffin-embedded sections of specimens of hypertrophic scars, keloids, normal skin, and surgical scar, and in situ hybridization for the expressions of IL-8, MCP-1, MIP-1alpha mRNA. RESULTS: The microvessel counts as well as the positive rates and the scorings of IL-8, MCP-1, and MIP-1alpha mRNA were significantly higher in pathological scars than those in the normal skin and surgical scar (all P < 0.05). The microvessel counts were significantly higher in the positive cases of IL-8, MCP-1 and MIP-1alpha mRNA than those in the negative ones (P < 0.05). The close positive correlations were found among the microvessel counts and the expressive scorings of 3 factors (P < 0.05). The close positive correlations were also found among the expressive scorings of IL-8, MCP-1, and MIP-1alpha mRNA in pathological scars. Microvessel counts were significantly higher in hypertrophic scars with the course less than 1 year than those with the course more than 1 year. CONCLUSION: IL-8, MCP-1 and MIP-1alpha play important roles in promoting the neovascularization of pathological scars. PMID- 16045030 TI - [Effect of diazepam and modafinil on acute hepatic failure in mice]. AB - OBJECTIVE: To investigate the effect of diazepam and modafinil on acute hepatic failure in mice. METHODS: Acute liver failure was induced in male Kunming strain mice by enterocoelia injecting the mice with D-GalN and LPS . The mice in the treatment groups were given corresponding drug 2 h before the administration of D GalN and LPS, and the mice in the control group were given the same dose of distilled water. The 24-hour survival rate, serum alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels were compared. Serum levels of TNF alpha and IL-1 and the levels of SOD, MDA, GR, GSH, NO and NOS in the liver were determined. RESULTS: Treatment with diazepam increased the survival rate and improved liver histological feature. Diazepam inhibited the serum levels of ALT, AST, TNF-alpha and IL-1, and reduced levels of MDA, NO and NOS and increased levels of GR and SOD in the liver. Modafinil decreased liver histological feature, increased the serum levels of ALT, AST, TNF-alpha and IL-1, increased level of MDA, and inhabited levels of SOD and GR in the liver. CONCLUSION: Treatment with diazepam may suppress the D-GalN/LPS-induced acute hepatic failure and modafinil may facilitate the acute hepatic failure. PMID- 16045031 TI - [Role of platelet activating factor in hepatorenal syndrome]. AB - OBJECTIVE: To investigate the relationship between platelet activating factor (PAF) and occurrence of hepatorenal syndrome (HRS). METHODS: A total of 57 cases of patients with liver cirrhosis or hepatitis gravis were enrolled. The serum concentrations of PAF, lipopolysaccharide (LPS) and prothrombin activity (PTA), total bilirubin, and serum creatinine were detected in patients combined with HRS and in patients without HRS in hospital at the term. The correlations among the PAF, LPS, and the function of liver and kidney were analyzed. RESULTS: The concentrations of PAF and LPS in the patients combined with HRS were significantly higher than those patients without HRS. There were linear correlations between the PAF and LPS level, the total bilirubin level and PAF level, the PAF level and serum creatinine level. CONCLUSION: The PAF associates with the occurrence of HRS. The PAF may mediate the occurrence of HRS induced by LPS. The PAF level may forecast whether patients with liver failure will take place HRS. PMID- 16045032 TI - [Genetics of ankylosing spondylitis]. PMID- 16045033 TI - [Conditional logistic model multivariate analysis on risk factors of nasopharyngeal carcinoma in Hunan]. PMID- 16045034 TI - [Early laying nasopharynx-duodenum tube for enteral nutrition during the patients undergoing the surgery of oesophagus of gastric cardia cancer]. PMID- 16045035 TI - [Methods for making high quality of frozen section for brain tissues]. PMID- 16045036 TI - [Loop electrosurgical excision procedure in the diagnosis and treatment of cervical intraepithelial neophlasia and early cervical carcinoma]. PMID- 16045037 TI - [Surgical treatment of 280 spontaneous hemopneumothorax patients]. PMID- 16045038 TI - [Diagnosis and treatment of thoracolumbar disc herniation]. PMID- 16045039 TI - [A case of dextroversion of heart combined with atrioventricular modal reentrant tachycardia]. PMID- 16045040 TI - [SCI and the instruction for submitting to its cited journals]. PMID- 16045042 TI - Triangulation. PMID- 16045043 TI - Illustrating triangulation in mixed-methods nursing research. AB - Graham R Williamson discusses the concept of triangulation in data collection and analysis in mixed-methods research, and reflects upon how this contributed to a deeper understanding when he used it in his work PMID- 16045044 TI - The use of triangulation for completeness purposes. AB - Triangulation in research refers to the use of multiple techniques for gathering and/or handling data within a single study The original purpose of triangulation was to seek confirmation of apparent findings. There now appears to be support for a second purpose - completeness. This paper reviews the arguments surrounding the use of triangulation, and shows how its use facilitated one researcher's attempt to provide a complete picture of the phenomenon explored in a qualitative study of the adequacy of the nurse education system in Malta. PMID- 16045045 TI - Methodological triangulation in midwifery education research. AB - This paper describes how the use of methodological triangulation can enrich the research process. The first section of the paper provides a brief outline of a national research project that studied 'pairs' of student midwives and their mentors in practice, and discusses the strengths and weaknesses of the approach. It then moves on to describe the combination of methods chosen for one aspect of the project before providing illustrative examples from the data that show how the triangulation of methods gave depth to the analysis PMID- 16045046 TI - Overseas black and minority ethnic nurses: a systematic review. AB - This paper is based on a study of overseas black and minority ethnic nurses in the NHS. Following a critical and analytical discussion of the principles of systematic reviewing, Obrey Alexis and Vasso Vydelingum describe the process of reviewing the literature and outlines the techniques of critically appraising all types of literature, including evidence that is opinion-based. PMID- 16045047 TI - Role conflict: appropriateness of a nurse researcher's actions in the clinical field. AB - This study aimed to judge the appropriateness of a particular nurse researcher's actions in a vignette from the clinical field, and to explore frameworks used to determine the appropriateness of actions. Twelve experienced nurse researchers were interviewed by telephone. They were given four vignettes of actual research situations and asked to comment and explain their judgement on the appropriateness of the nurse researcher's actions. The findings confirmed blurring of boundaries between professional and research roles for nurses. Nurses appear to use a mixture of frameworks, including scientific/rigour method, ethics, nursing competencies and their own personal-moral values. Frameworks need to be discussed and debated so that neophyte researchers are well-prepared before they enter the field to conduct research. PMID- 16045048 TI - Balance of power--do patients use researchers to survive hospital? AB - This paper focuses on the way research study participants interpret their contact with researchers. Much has been made of the unequal power relationship between researcher and researched, but the case study presented here suggests that patient participants may be 'using' researchers to further their own agenda and to attempt to manipulate or control aspects of their care within the hospital setting. PMID- 16045049 TI - 'Research positively embraces questions and challenges'. AB - RCN general secretary Beverly Malone recalls a former colleague and mentor, and explains why, in the belief that research is integral in helping to shape and deliver care, she continues to campaign hard for nurses to have three days every year for continuing professional development. PMID- 16045050 TI - Interdisciplinary and intradisciplinary nursing education: none, either or both? PMID- 16045051 TI - Towards an "open and frank dialogue". PMID- 16045052 TI - Joan Shaver on nurturing leaders. PMID- 16045053 TI - Calling all leaders. PMID- 16045054 TI - Interprofessional education for collaborative, patient-centred practice. PMID- 16045055 TI - Patient safety: is it just another bandwagon? AB - In this paper, the author questions the focus of the patient safety movement, speculating that it might be just another "bandwagon" that health executives and some health professionals are eager to join. The history of this current emphasis on patient safety is briefly sketched, including current activities in Canada, and questions are raised about whether the movement aids or avoids pressing healthcare issues, many of which are supported by good evidence. These include the relationship between nursing staffing and patient outcomes, the way in which a "cult of efficiency" has operated to make errors more likely and how the silencing of nurses and other staff leads to error. Whether or not one considers the current focus on patient safety a bandwagon, it is important to reflect critically upon the activities undertaken to address the issue and to determine whether one should jump on this bandwagon. PMID- 16045056 TI - Canadian nursing in the genomic era: a call for leadership. AB - With developments in genomics, there is an urgent need for Canadian nurses in all settings to be informed and involved in the incorporation of this new knowledge into healthcare. The purpose of this paper was to synthesize the literature on genetic nursing roles to provide a foundation for Canadian nursing leaders as they take on the challenges of nursing in the genomic era. A comprehensive review of 98 articles published between 1994 and 2004 revealed strong support for genetic nursing roles and recommendations for levels of genetic nursing practice. The few studies on genetic nursing roles suggested that nurses and other health professionals support the integration of genomics into nursing practice. Issues that need to be addressed related to nursing practice, education and research are offered to engage nursing leaders in advocating for the incorporation of genomics into nursing in Canada. PMID- 16045057 TI - Creating and sustaining dementia special care units in rural nursing homes: the critical role of nursing leadership. AB - Dementia Special Care Units (SCUs) are more likely to be found in larger nursing homes, which tend to be located in urban centres, rather than in smaller rural nursing homes. Reasons for the small number of rural SCUs are not known, although it has been speculated that space and staffing constraints, lack of a critical mass of residents needing specialized care and limited resources may be important factors. The purpose of this study was to describe the development of SCUs in eight small rural nursing homes (31-100 beds) in Saskatchewan, Canada, from the perspective of nursing directors involved in planning and implementing the units. Although the initial focus was on how and why the SCUs were established, the key finding was the critical role of nursing leadership and supervision in creating and sustaining the unit. Even the most successful SCUs required constant vigilance to maintain an effective program, highlighting their inherent fragility and the need for a designated, committed leader. Four key leadership activities were identified: perpetual reinforcement and enforcement of SCU goals and ideals; support, guidance and mentoring of staff; empowerment of staff; and liaison/public relations. PMID- 16045058 TI - Court upholds New Jersey Medical Board's regulation affecting CRNAs. PMID- 16045059 TI - Texas court affirms summary judgment in suit by nurse. PMID- 16045060 TI - Chronotherapeutics (light and wake therapy) in affective disorders. AB - The Committee on Chronotherapeutics, delegated by the International Society for Affective Disorders (ISAD), makes the following recommendations after reviewing the evidence as of November 2004. (1) Wake therapy is the most rapid antidepressant available today: approximately 60% of patients, independent of diagnostic subtype, respond with marked improvement within hours. Treatment can be a single or repeated sleep deprivation, total (all night) or partial (second half of the night). Relapse can be prevented by daily light therapy, concomitant administration of SSRIs, lithium (for bipolar patients), or a short phase advance of sleep over 3 days following a single night of wake therapy. Combinations of these interventions show great promise. (2) Light therapy is effective for major depression--not only for the seasonal subtype. As an adjuvant to conventional antidepressants in unipolar patients, or lithium in bipolar patients, morning light hastens and potentiates the antidepressant response. Light therapy shows benefit even for patients with chronic depression of 2 years or more, outperforming their weak response to drugs. This method provides a viable alternative for patients who refuse, resist or cannot tolerate medication, or for whom drugs may be contraindicated, as in antepartum depression. (3) Given the urgent need for new strategies to treat patients with residual depressive symptoms, clinical trials of wake therapy and/or adjuvant light therapy, coupled with follow-up studies of long-term recurrence, are a high priority. PMID- 16045061 TI - Controlled trial of bright light and negative air ions for chronic depression. AB - BACKGROUND: This randomized controlled trial investigates the efficacy of two non pharmacologic treatments, bright light and high-density negative air ions for non seasonal chronic depression. Both methods have shown clinical success for seasonal affective disorder (SAD). METHOD: Patients were 24 (75%) women and 8 (25%) men, ages 22-65 years (mean age +/- S.D., 43.7 +/- 12.4 years), with Major Depressive Disorder, Single Episode (DSM-IV code, 296.2), Chronic (episode duration > or = 2 years). Patients were entered throughout the year and randomly assigned to exposure to bright light (10 000 lux, n = 10), or high-density (4.5 x 10(14) ions/s flow rate, n = 12) or low-density (1.7 x 10(11) ions/s, n = 10, placebo control) negative air ions. Home treatment sessions occurred for 1 h upon awakening for 5 weeks. Blinded raters assessed symptom severity weekly with the Structured Interview Guide for the Hamilton Depression Rating Scale--Seasonal Affective Disorder (SIGH-SAD) version. Evening saliva samples were obtained before and after treatment for ascertainment of circadian melatonin rhythm phase. RESULTS: SIGH-SAD score improvement was 53.7% for bright light and 51.1% for high density ions v. 17.0% for low-density ions. Remission rates were 50%, 50% and 0% respectively. The presence or severity of atypical symptoms did not predict response to either treatment modality, nor were phase advances to light associated with positive response. CONCLUSIONS: Both bright light and negative air ions are effective for treatment of chronic depression. Remission rates are similar to those for SAD, but without a seasonal dependency or apparent mediation by circadian rhythm phase shifts. Combination treatment with antidepressant drugs may further enhance clinical response. PMID- 16045062 TI - The effects on suicide rates of an educational intervention for front-line health professionals with suicidal patients (the STORM Project). AB - BACKGROUND: The opportunity to study district-wide educational interventions on suicide rates is rarely available. In 1997, the authors carried out a district wide training programme for primary care, accident and emergency, and mental health workers (47% of eligible staff trained), and demonstrated improvements in skills, attitude and confidence among the recipients of the training. METHOD: Suicide rates (including definite suicides and undetermined deaths) and population statistics were collected for a district and region of England from official sources from 1993-2001. A before-and-after (1994-1996 and 1998-2000) training intervention analysis was conducted on suicide rates. RESULTS: The suicide rate in 1994-1996 was 8.8 per 100 000 before our educational intervention and unchanged at 8.6 per 100 000 in 1998-2000 after it (p = 0.783). CONCLUSION: Brief educational interventions to improve the assessment and management of suicide for front-line health professionals in contact with suicidal patients may not be sufficient to reduce the population suicide rate. PMID- 16045063 TI - Improving traditional intention-to-treat analyses: a new approach. AB - BACKGROUND: Drop-out, often accompanied by treatment non-compliance, is common in psychiatric trials. Methodologists have criticized the use of a traditional intention-to-treat (ITT) approach in such cases, and have proposed alternative methods. We set out to describe and assess methods for estimation of a treatment effect when the trial is 'broken'. METHOD: We describe a stratified method of moments (SMOM) estimator that assesses treatment effects on subjects who are willing to comply with all the treatments under study. A simulation study and a re-analysis of data from an antipsychotics trial are used to compare SMOM to ITT, as-treated, and adequate estimators. RESULTS: The new estimator retains good statistical properties under different levels of non-compliance and drop-out mechanisms. The re-analysis indicates that SMOM yields more precise results. CONCLUSIONS: Although the traditional ITT approach provides a valid method to estimate treatment effects, it can be biased in the presence of treatment non compliance and drop-out. It is critical that researchers move beyond traditional approaches when trials are broken. A key first step is to consider non-compliance and drop-out as two independent phenomena, tracking and reporting rates separately. PMID- 16045064 TI - Sexual orientation and mental health in a birth cohort of young adults. AB - BACKGROUND: This paper sought to examine the relationship between sexual orientation and mental health in a New Zealand birth cohort studied to age 25 years. METHOD: The analysis is based on a sample of 967 participants (469 males; 498 females) in the Christchurch Health and Development Study. As part of this study information was gathered on: (a) measures of sexual orientation, same-sex behaviour and sexual attraction obtained at ages 21 and 25 years; (b) measures of mental disorders and suicidal behaviours over the interval 21-25 years; (c) measures of childhood and family background. RESULTS: Latent class analysis was used to combine indicators of sexual orientation, same sex behaviour and attraction to form an empirically based classification of sexual orientation. The best-fitting model classified the sample into three groups: exclusively heterosexual orientation (87.6%); predominantly heterosexual but with same-sex inclinations or experience (9.6%); predominantly homosexual (2.8%). Proportionately more women than men were classified as predominantly heterosexual (14.2% v. 4.8% respectively) or predominantly homosexual (3.9% v. 1.5% respectively). Cohort members with a predominantly homosexual orientation had rates of mental disorder and suicidal behaviours that were between 1.5 and 12 times higher than for those with an exclusively heterosexual orientation. These associations persisted after adjustment for confounding. The associations between sexual orientation and mental health were more marked for males than females. CONCLUSIONS. The findings suggest a continuum of sexual preferences amongst young adults. Variations in sexual orientation were clearly associated with mental health. These associations tended to be stronger for males. PMID- 16045065 TI - Suicidal behaviour in adolescence and subsequent mental health outcomes in young adulthood. AB - BACKGROUND: The aim of this study was to examine the linkages between suicidal ideation and attempt in adolescence and subsequent suicidal behaviours and mental health in young adulthood. METHOD: Data were gathered during the course of a 25 year longitudinal study of a birth cohort of 1265 New Zealand children. The information collected included: (a) measures of suicidal thoughts and attempts in adolescence (< 18 years); (b) measures of suicidal ideation, suicide attempt, major depression, anxiety disorders, and substance use disorders in young adulthood (18-25 years); and (c) measures of childhood and family background, individual characteristics, and mental disorders in adolescence. RESULTS: After statistical adjustment for confounding factors, suicide attempt in adolescence was associated with increased risks of subsequent suicidal ideation (OR 5.7) suicide attempt (OR 17.8) and major depression (OR 1.5). Those reporting suicidal ideation without suicide attempt showed moderate increases in risks of later suicidal ideation (OR 2.5), suicide attempt (OR 2.0) and major depression (OR 1.6). In addition, there was evidence of an interactive relationship in which suicidal behaviour in adolescence was associated with increased risks of later substance use disorders in females but not males. CONCLUSIONS: Young people reporting suicidal ideation or making a suicide attempt are an at-risk population for subsequent suicidal behaviour and depression. Further research is needed into the reasons for suicidal adolescent females being at greater risk of later substance use disorder. PMID- 16045066 TI - Stability and clinical correlates of personality characteristics in recent-onset schizophrenia. AB - BACKGROUND: While several personality characteristics are implicated in the etiology and phenotypic expression of schizophrenia, little is known about their trait status or clinical correlates during the early course of this disorder. METHOD: The stability of five personality characteristics derived from the MMPI 168 was examined in recent-onset schizophrenia patients (n = 59) during a psychotic period at study entry and at two out-patient assessments over the following 15 months and in non-patient controls (n = 39) at similar time intervals. Among patients, associations between personality characteristics and clinical symptoms and exposure to stressful life events during the study period were also examined. RESULTS: Multilevel modeling analyses indicated that the between-subjects or trait-like variance components for these characteristics among patients were substantial, albeit smaller than those found for controls. Patients reported higher mean levels of Neuroticism, Cynicism, and Psychotic Ideation and lower mean levels of Psychopathic Tendencies and Denial of Somatic Complaints than controls. Among patients, several personality characteristics were systematically related to mean clinical symptom levels as well as frequency of exposure to dependent, but not independent, life events. CONCLUSIONS: Stable individual differences in personality are detectable during the early course of schizophrenia and may help account for heterogeneity in clinical symptoms and functional outcome. PMID- 16045067 TI - Cognitive control in childhood-onset obsessive-compulsive disorder: a functional MRI study. AB - BACKGROUND: Failure to resist chronic obsessive-compulsive symptoms may denote an altered state of cognitive control. We searched for the cerebral regions engaged in this dysfunction. METHOD: Differences in brain regional activity were examined by event-related functional magnetic regional imaging (fMRI) in a group of adolescents or young adults (n = 12) with childhood-onset obsessive-compulsive disorder (OCD), relative to healthy subjects. Subjects performed a conflict task involving the presentation of two consecutive and possibly conflicting prime and target numbers. Patients' image dataset was further analysed according to resistance or non-resistance to symptoms during the scans. RESULTS: Using volume correction based on a priori hypotheses, an exploratory analysis revealed that, within the prime-target repetition condition, the OCD subjects activated more than healthy subjects a subregion of the anterior cingulate gyrus and the left parietal lobe. Furthermore, compared with 'resistant' patients, the 'non resistant' OCD subjects activated a bilateral network including the precuneus, pulvinar and paracentral lobules. CONCLUSIONS: Higher regional activations suggest an abnormal amplification process in OCD subjects during the discrimination of repetitive visual stimuli. The regional distribution of functional changes may vary with the patients' ability to resist obsessions. PMID- 16045068 TI - Functional MRI of visuospatial working memory in schizotypal personality disorder: a region-of-interest analysis. AB - BACKGROUND: Functional MRI studies have begun to identify neural networks implicated in visuo-spatial working memory in healthy volunteers and patients with schizophrenia. The study of schizotypal personality disorder (SPD) provides regional analysis in unmedicated patients in the schizophrenia spectrum. METHOD: Unmedicated patients with SPD by DSM-IV criteria and normal controls were assessed with fMRI while performing a visuo-spatial working-memory task. It required the subjects to retain the location of three dots located on the circumference of an imaginary circle and then respond to a query display in which one dot was presented and the subject required to press a button to indicate whether the probe dot location was previously displayed. Subject groups did not differ significantly in spatial memory scores. The exact Talairach and Tournoux coordinates of brain areas previously reported to show activation with spatial memory tasks were assessed. RESULTS: The majority of these locations showed BOLD response activation significantly less in patients during the memory retention period, including the left ventral prefrontal cortex, superior frontal gyrus, intraparietal cortex and posterior inferior gyrus. Regions in the right middle prefrontal and prestriate cortex showed greater activation at a trend level for patients with SPD than for normal controls. In addition, we replicated the findings of increased activation with the task in healthy volunteers in the premotor areas, ventral prefrontal cortex and parietal cortex. CONCLUSIONS: SPD patients show decreased activation compared to healthy volunteers in key frontal regions and we also provided a partial replication of findings reported in healthy subjects. PMID- 16045069 TI - Visuospatial learning and executive function are independently impaired in first episode psychosis. AB - BACKGROUND: Demonstrating specific cognitive impairments in psychotic disorders is difficult. However, specific deficits in memory and executive functions have often been claimed. The Cambridge Neuropsychological Test Automated Battery (CANTAB) tasks of IDED attention-shifting (an executive task) and visuospatial paired associates learning (PAL, a memory task) require intact frontal and temporo-hippocampal functioning, respectively; both have been suggested as markers of disease progress in psychosis. METHOD: Seventy-one subjects with a first-episode psychosis or at-risk mental state were assessed on these two tasks during referral to a specialist service, the Cambridge-based CAMEO early intervention team. RESULTS: Performance on the two tasks was dissociated. Poor performance on the PAL test was associated with increased symptom levels and poorer global function, while failure on the IDED executive test was not found to have significant clinical associations. Duration of illness was not associated with performance on either task. CONCLUSIONS: Visuospatial PAL failure may be a marker of clinical severity at the onset of psychosis while IDED performance may reflect a more stable, trait-like impairment. Dissociated performance on the executive and associative learning tasks may reflect independent, neurally dissociated impairments that do not arise in a fixed order. This may explain some of the heterogeneity of cognitive function seen in early psychosis. PMID- 16045070 TI - Lateral interactions in the visual cortex of patients with schizophrenia and bipolar disorder. AB - BACKGROUND: Schizophrenia is associated with perceptual organization deficits and abnormal neuronal connectivity has been described in early visual areas. The purpose of this study was to investigate the functional integrity of lateral connections in early visual areas of patients with schizophrenia and type I bipolar disorder with a history of psychosis. METHOD: Twenty-four out-patients with schizophrenia, 22 out-patients with bipolar disorder, and 20 healthy control subjects participated in the study. Using a computer-assisted psychophysical test, contrast thresholds were measured for centrally presented target stimuli (Gabor patches), which were surrounded by two collinear flankers. Target-to flanker distances were 0, 1, 2, 3, 4, 6, 9 and 122. Psychophysical measures were contrast threshold changes at each target-to-flanker distance compared with baseline thresholds determined for isolated targets with no flankers. Clinical measures included IQ, positive, negative, and depressive symptoms. Results. In patients with schizophrenia, flankers did not facilitate contrast detection for target stimuli at 2-6 lambda distances compared to controls [effect size (Cohen's d): 1.25-1.42]. The inhibitory effect of flankers (0 and 1lambda) and contrast thresholds in the absence of flankers were spared. Patients with bipolar disorder did not differ from the controls. Medicated and non-medicated patients displayed similar performances. Positive and negative symptoms and depression did not correlate with contrast threshold values. CONCLUSIONS: Excitatory lateral connections in early visual cortex are specifically impaired in patients with schizophrenia, which may contribute to perceptual disorders such as unclear seeing, partial or skewed sight, disrupted rectilinearity, and abnormal figure ground segregation. PMID- 16045071 TI - Early processing deficits in object working memory in first-episode schizophreniform psychosis and established schizophrenia. AB - BACKGROUND: While there are many studies showing working-memory deficits in schizophrenia there are only a few that disentangle impairments for working memory subprocesses such as perceptual, attentional, mnemonic and executive function. METHOD: In this study of delay-dependent memory, 55 patients with schizophreniform psychosis, 50 with established schizophrenia and 56 healthy controls were investigated. Using the delayed matching-to-sample task from the Cambridge Neuropsychological Test Automated Battery (CANTAB), performance deficits were found in both patient groups after controlling for age and pre morbid IQ. RESULTS: Even after controlling for simultaneous matching-to-sample ability (i.e. perceptual matching), impaired performance in both patient groups was found as soon as the stimuli were no longer present. Impaired performance was not due to different types of errors in patients versus controls. Performance in both patient groups was comparable, except for a slight decrease of overall task performance. This suggests that the deficit is relatively stable during the course of the illness. CONCLUSIONS: Our results suggest a deficit in patients with psychotic illness in the initial processes necessary to actively maintain information, such as the ability to form an internal representation of complex objects. PMID- 16045072 TI - The development and internal consistency of the satisfaction with Antipsychotic Medication Scale. AB - BACKGROUND: Satisfaction with antipsychotic medication is an important outcome variable. To date, there is a lack of a well-established measure to quantify patient satisfaction with psychiatric medication. This paper describes the development, dimensionality, reliability and validity of the Satisfaction with Antipsychotic Medication (SWAM) scale. METHOD: Clinical and academic experts devised a 33-item Likert scale satisfaction questionnaire. Following a pilot study in a sample of 69 people with schizophrenia, 315 people with schizophrenia on the caseload of local mental health services in three London boroughs completed the questionnaire. The dimensionality, internal consistency and validity of the devised instrument were assessed. RESULTS: Reliability of the SWAM scale was good for subscales and total scores. The alpha coefficient for the two subscales: treatment acceptability and medication insight were 0.92 and 0.84 respectively. The a coefficient for the SWAM scale total score was 0.91 and ranged from 0.92 to 0.90. CONCLUSION: Testing of the psychometric properties of the SWAM scale demonstrate that it is a reliable instrument for measuring patient satisfaction with antipsychotic medication. The measure could be used in routine clinical practice in mental health services to assess patient satisfaction with psychiatric medication. PMID- 16045073 TI - Rethinking the duration requirement for generalized anxiety disorder: evidence from the National Comorbidity Survey Replication. AB - BACKGROUND: The proposed revisions of the ICD and DSM diagnostic systems have led to increased interest in evaluation of diagnostic criteria. This report focuses on the DSM-IV requirement that episodes of generalized anxiety disorder (GAD) must persist for at least 6 months. Community epidemiological data are used to study the implications of changing this requirement in the range 1-12 months for estimates of prevalence, onset, course, impairment, co-morbidity, associations with parental GAD, and sociodemographic correlates. METHOD: Data come from the US National Comorbidity Survey Replication (NCS-R), a US household survey carried out during 2001-2003. Version 3.0 of the WHO Composite International Diagnostic Interview (WMH-CIDI) was used to assess DSM-IV anxiety disorders, mood disorders, substance disorders, and impulse-control disorders. RESULTS: Lifetime, 12-month, and 30-day prevalence estimates of DSM-IV GAD changed from 6.1%, 2.9%, and 1.8% to 4.2-12.7%, 2.2-5.5%, and 1.6-2.6% when the duration requirement was changed from 6 months to 1-12 months. Cases with episodes of 1-5 months did not differ greatly from those with episodes of > or = 6 months in onset, persistence, impairment, co-morbidity, parental GAD, or sociodemographic correlates. CONCLUSIONS: A large number of people suffer from a GAD-like syndrome with episodes of < 6 months duration. Little basis for excluding these people from a diagnosis is found in the associations examined here. PMID- 16045074 TI - Comparison of the CIS-R and CIDI lay diagnostic interviews for anxiety and depressive disorders. PMID- 16045075 TI - A rearview reflection for pragmatic planning. PMID- 16045076 TI - Strategic planning and information technology: back to the future all over again. PMID- 16045077 TI - Show strategies, plans, and strategic planning. PMID- 16045078 TI - Playing Post Office with healthcare. PMID- 16045079 TI - The new alphabet soup. PMID- 16045080 TI - RFID: it's about more than asset tracking. PMID- 16045081 TI - Successful strategic planning: creating clarity. AB - Most healthcare organizations have a strategic plan of some kind. Many of these organizations also have difficulty translating their strategic plan into specific actions that result in successful performance. In the worst cases, this can jeopardize the viability of the organization. The trouble lies in a lack of clarity in what a strategic plan is and what it should do for the organization. This article will answer key questions such as: What is strategy and how does it fit with other commonly used constructs such as mission, vision, and goals? What criteria can be used to determine if something is truly strategic to the organization? What are the phases of the strategy lifecycle? How do approaches for dealing with uncertainty, such as scenario planning, fit with organizational strategic planning? How can a meaningful IT strategy be developed if the organization strategy is lacking? What principles should guide a good IT planning process? PMID- 16045082 TI - Information technology strategic planning: art or science? AB - It had been almost a decade since the hospitals that make up the Daughters of Charity Health System (DCHS) had engaged in a formal information technology strategic planning process. In the summer of 2002, as the health system re formed, there was a unique opportunity to introduce a planning process that reflected the governance style of the new health system. DCHS embarked on this journey, with the CIO initiating and formally sponsoring the information technology strategic planning process in a dynamic and collaborative manner The system sought to develop a plan tailored to encompass both enterprise-wide and local requirements; to develop a governance model to engage the members of the local health ministries in plan development, both now and in the future; and to conduct the process in a manner that reflected the values of the Daughters of Charity. The DCHS CIO outlined a premise that the CIO would guide and be continuously involved in the development of this tailored process, in conjunction with an external resource. Together, there would be joint responsibility for introducing a flexible information technology strategic planning methodology; providing an education on the current state of healthcare IT, including future trends and success factors; facilitating support to tap into existing internal talent; cultivating a collaborative process to support both current requirements and future vision; and developing a well-functioning governance structure that would enable the plan to evolve and reflect user community requirements. This article highlights the planning process, including the lessons learned, the benchmarking during and in post-planning, and finally, but most importantly, the unexpected benefit that resulted from this planning process. PMID- 16045083 TI - The development and implementation of a strategic and tactical planning framework at Mayo Clinic Arizona. AB - This article describes the Mayo Arizona process for developing an information technology strategic plan. The background of organizational events that gave rise to this strategic planning process is presented. A cross-functional team of key IT stakeholders was convened; the team used a facilitated process to derive a pro forma set of IT strategic objectives from the larger organization's emerging strategic plan. A broad set of leadership interviews was conducted to further identify detailed objectives that would confirm, complement, or conflict with the "strawperson." The IT strategic objectives then were refined and published by the organization. The article also describes the annual process of reviewing the IT strategic plan and translating it to a set of tactical objectives. This includes the committee structure for project prioritization, which is guided by the IT strategic plan. The outcome of the prioritization process is a five-year IT tactical plan, which is used to communicate the IT action plan for achievement of the strategic objectives. The strategic and tactical plans have resulted in stronger ownership and advocacy of IT activities by organizational leadership and a clearer view of the impact of technology on the organization's strategic plan. PMID- 16045084 TI - Developing and implementing a patient-centered IT strategy. AB - The Army Medical Department (AMEDD), a comprehensive worldwide integrated healthcare system with an annual budget of more than $6 billion, more than 50,000 employees, and 2.4 million beneficiaries, developed and implemented a comprehensive patient-centered enterprise-wide information management and information technology strategy to facilitate information management systems and infrastructure decisions by leaders. This article describes a patient-centered model used to organize and link healthcare activities and activity leaders to portray patient care, administrative, business, financial, supply, and strategic support information systems. Activity and IT leaders applied a refined strategic alignment model to identify specific clinical, business, and IT goals and to detail the necessary infrastructure investments using a systems view. The use of patient and process outcome measures tied to the AMEDD's Balanced Score Card' helped leaders to manage IT strategy execution. Now, two years into the effort, a sample activity strategy--outpatient care--is used to illustrate the application of these tools to the development and implementation of a patient-centered IT strategy. PMID- 16045085 TI - The planning cycle. AB - Information technology planning can be described as a continuous cyclical process composed of three phases whose primary purpose is optimum allocation of scarce resources. In the assessment phase, planners assess user needs, environmental factors, business objectives, and IT infrastructure needs to develop IT projects that address needs in each of these areas. A major goal of this phase is to develop a broad IT inventory. The prioritization phase seeks to ensure optimum allocation of scarce resources by prioritizing ITprojects based on: Costs--total life cycle costs. Benefits--both quantitative and non-quantitative, including support for the organization's strategic business objectives. Risks--subjective assessments of technological and non-technological risks. Implementation requirements--time and personnel requirements to implement the system. The scheduling phase incorporates sequencing considerations, personnel availability, and budgetary constraints to produce an IT plan in which project priorities are adjusted to meet organizational realities. PMID- 16045086 TI - Patient note deidentification using a find-and-replace iterative process. AB - Medical research relies on access to clinical data. Health Insurance Portability and Accountability Act regulations require that patient information required for clinical research not have data that can be used to identify the patients from whose medical records the information has been derived. The only exception would be an institutional review board (IRB)-approved study for which the researcher has obtained a waiver to use patient data for a research study. Before requesting an IRB waiver, however, the researcher may want to search the clinical data for particular characteristics or determine whether the quantity of data warrant obtaining IRB approval. The application, the Simple PAtient Note Scrubber, or SPANS, reviews and changes line content through an iterative process. At each iteration, SPANS analyzes changes made during the previous pass and reviews changes in relation to terms adjacent to the newly altered data. Knowledge of the document domain, encompassing the different types of documents to be scrubbed, is the key to making this type of process effective. PMID- 16045087 TI - Human factors considerations relevant to CPOE implementations. AB - Although many studies have discussed the benefits of computerized provider order entry (CPOE), the actual number of hospitals using this technology remains low because of the many challenges that accompany the implementation of CPOE in healthcare facilities. It is common for user resistance to challenge implementation efforts. As more hospitals undertake CPOE implementations, a solid understanding of how to foster acceptance of CPOE is necessary to reap the benefits of medical error reduction, improved quality of care, and decreased healthcare costs. The principles and practices of human factors can be used to bolster physician satisfaction and increase usability, thereby increasing the chances of success for CPOE implementation. This article reviews the recent literature regarding CPOE and human factors, discussing how the human factors principles of task analysis, interface design, and computer supported cooperative work can be utilized to promote user acceptance and enhance CPOE implementation efforts. PMID- 16045088 TI - Ponderal index. PMID- 16045089 TI - Ponderal index of low birth weight babies--a hospital based study. AB - OBJECTIVE: To determine the ratio of disproportionate versus proportionate Intra uterine growth retardation among low birth weight babies using the Ponderal index. METHODS: This was a retrospective study. Medical records of babies with weight < 2.5kg delivered in the Obstetric Department were reviewed. The ponderal index was calculated to determine disproportionate and proportionate intra uterine growth retardation among low birth weight babies. RESULTS: A total of 227 babies were included. Male and Female ratio was equal. Sixty seven percent were delivered vaginally. Only 12 mothers (5.3%) had a history of maternal illness. Eighty nine low birth weight babies (39.9%) had a low ponderal index. Ponderal index in fullterm babies was < 2.2 in 54.3% and in pre-term < 2.0 in 34.9%. CONCLUSION: Nearly 40% of low birth weight babies had disproportionate body proportions. This group of babies according to foetal origins of diseases may be predisposed to Diabetes, hypertension and coronary artery disease. PMID- 16045090 TI - Rotavirus infection in infants and young children in Makkah, Saudi Arabia. AB - OBJECTIVE: To determine the prevalence of rotavirus in infants and young children in Makkah, Saudi Arabia. METHODS: A population-based prevalence study was done in randomly selected infants and young children suffering from acute diarrhoea. Faecal specimens were collected from 479 patients. A latex agglutination test was used for rotavirus detection. All positive and 10 negative samples for rotavirus by latex agglutination were also tested by enzyme-linked immunosorbent assay (ELISA). RESULTS: Rotavirus was detected in 48 (10%) patients using latex agglutination test. ELISA detected 46/48 positive samples. Ten samples that tested negative with latex test were also negative with ELISA. Infection with rotavirus was more frequent among infants and children < 2 years old, with a maximum incidence among children 0-12 months. Rotavirus infection was 3.1% in Saudi nationals, compared to 6.9% in other nationalities. CONCLUSION: In this study the prevalence rate of 10% was low compared to other studies done in different regions of Saudi Arabia. This low rate could be due to the geographical location of Makkah with very hot and dry summer, and mild winter and almost no rain throughout the year. PMID- 16045091 TI - FLAG-IDA in the treatment of refractory/relapsed acute leukaemias: single centre study. AB - OBJECTIVE: To evaluate the efficacy and toxicity profile of the combination of fludarabine, high dose cytarabine, idarubicin, and granulocyte colony stimulating factor in refractory relapsed cases of acute leukaemia, a study is being conducted at Armed Forces Bone Marrow Transplant Centre (AFBMTC) Rawalpindi since January 2003. Data up to June 2004 (early report) is being presented. METHODS: Twelve Patients with refractory/relapsed (Ref/Rel) acute leukaemia (AL) were treated with fludarabine 30 mg/m2 and cytosine arabinoside (AraC) Arac 2 g/m2 for 5 days, idarubicin 10 mg/m2 for 3 days, and granulocyte colony stimulating factor G-CSF 5 micro g/kg from day 0 till neutrophil recovery (ANC > 1.0 x 10(9)/1). Response was evaluated by bone marrow examination on day 20-post chemotherapy. RESULTS: Patients included were refractory acute lymphoblastic leukaemia (ALL) (n=2), relapsed ALL (n = 3), refractory acute myeloid leukaemia (AML) (n = 3), secondary AML (n=2) relapsed AML (n = 1) and acute undifferentiated leukaemia (AUL) (n = 1). Complete remission (CR) was achieved in 8 (66.6%) patients. Three (25%) patients died of post chemotherapy complications and one patient failed to achieve remission. Out of 8 patients who achieved CR, 4 underwent allogeneic bone marrow transfusion (BMT), 1 is being evaluated for the same, 1 received idorubicin, AraC and etopuside (ICE) and high dose AraC, 1 did not receive further chemotherapy and 1 relapsed two months after remission. Seven patients are still in CR after a median follow up of 8 months (range 3-18). Major complications encountered were diarrhoea, mucositis, toxic ileus, transient hepatic toxicity, fungal and bacterial infections. CONCLUSION: In our experience, FLAG-IDA is well tolerated and effective regimen in relapsed/refractory acute leukaemias. The toxicity is acceptable, enabling most patients to receive further treatment, including transplantation procedures. PMID- 16045092 TI - To compare the total cholesterol and HDL-cholesterol before and after ultra centrifugation in lipemic samples. AB - OBJECTIVE: It is an everyday routine in laboratories to encounter interference in the analysis of lipids. These likely interferences (hemolysis, icterus and lipemia) are countered by asking physicians and patients to send fresh and properly collected samples. At the Aga Khan University (AKU) Laboratory, we receive 2-3 lipemic/turbid samples per day. Previously it was our departmental policy to advise these patients to go for lipoprotein electrophoresis, which though accurate was time consuming and not cost effective. We therefore studied ultra centrifugation/airfuge as an alternate method to clear lipid interference and provide accurate, reliable and cost effective results. METHODS: Daily 2-3 grossly turbid samples are identified on the lipid bench, 48 samples were received in 4 months (February to May 2004). These samples were analyzed for total cholesterol (TC) and High Density Lipoprotein-Cholesterol (HDL-C) before and after ultra centrifugation/airfuge. RESULTS: There was a positive correlation between the lipemia and the false high TC and HDL-C. The mean TC and HDL-C before ultra centrifugation were 263.06 mg/dl and 39.42 mg/dl respectively and after centrifugation these became 191.77 mg/dl and 33.06 mg/dl. P value showed a significant difference in both results. CONCLUSION: This study suggests that the removal of turbidity by ultra centrifugation/airfuge is cost effective, less time consuming and provides accurate reliable results of TC and HDL-C in patients with lipemia interference. PMID- 16045093 TI - Vaginitis and sexually transmitted infections in a hospital based study. AB - OBJECTIVE: To determine the frequency of Sexually transmitted infections (STIs) in immigrant population versus the local population, comparing the symptomatic patients against the asymptomatic women as controls. METHODS: A cross-sectional comparative study was conducted in Gynaecology Clinic of Unit 1, Bolan Medical Complex Hospital, Quetta. The study duration was six months from 1st April, 2004 to 31st October, 2004, examining 500 women. World Health Organization (WHO) recommended questionnaire was answered and High Vaginal Swab (HVS) was taken from posterior fornix of patient's vagina. Gram staining was done by the Pathology Department of the same hospital and description was based on Nugent's score and modified Spiegel's criteria. Blood was analyzed for human immunodeficiency virus (HIV) and hepatitis-B surface antigen (HBsAg) by immunochromatography. Analysis of results was done by Chi square test, screening test validity and incidence testing. RESULTS: Women with vaginal discharge had higher incidence of STI (84%) than without it (38%). Refugees and women with grand multiparity made the high risk groups of targeted testing (P < 0.001). The commonest notifiable infections, in this study were vaginitis (33.48%), bacterial vaginosis (30.7%), candidiasis (10%), trichomoniasis (7.2%), gonorrhea (1.35%) and 1 case each of lymphogranuloma venereum and chancroid, and 3 carrier states of HbsAg were detected. No case of syphilis or HIV/AIDS, or genital herpes was found. CONCLUSION: This study will serve as a marker for the tip of an iceberg of STI in Balochistan, needing targeted testing in high risk groups. PMID- 16045094 TI - Frequency of thrombophilia in patients with adverse pregnancy outcome. AB - OBJECTIVE: To determine the association between adverse pregnancy outcomes and thrombophilia. METHODS: This is a descriptive study, incorporating retrospective analysis of patients with recurrent pregnancy losses, intrauterine deaths, abruptio placenta and early onset pre eclampsia. Patients with adverse pregnancy outcomes in whom co-morbid factors were excluded underwent screening for both acquired and inherited thrombophilia. RESULTS: A total of 40 patients were screened for acquired and inherited thrombophilia with adverse pregnancy outcomes. Anticardiolipin antibodies were found positive in 55% of patients and 45% of patients were found deficient for natural anticoagulants protein C and S. Two patients were found positive for both acquired and inherited thrombophilia. CONCLUSION: Thrombophilia, both acquired and inherited are associated with adverse pregnancy outcomes. Patients in whom other co-morbid factors are excluded, should be offered screening for thrombophilia. Liaison between hematologist and obstetrician is the corner stone for success. PMID- 16045095 TI - Surgically treated rectal prolapse: experience at a teaching hospital. AB - OBJECTIVE: To analyze our results regarding the surgical treatment of complete rectal prolapse performed at the Aga Khan University Hospital, from January 1988 to December 2003. METHODS: Files were retrieved from our medical records and data was reviewed for all adult patients admitted and operated upon for complete rectal prolapse during our study period. Long-term follow-ups were obtained through these files and also by contacting patients through telephone and letters. Data was recorded in a standardized two-page proforma and analysis was carried out between different variables using SPSS 10.0. RESULTS: A total number of twenty surgeries (n = 20) were performed. All patients had the presenting complaint of something coming out of anus, 70% (n = 14) patients complained of some bleeding per rectum, 30% (n = 6) had anal pain and 20% (n = 4) had faecal incontinence. Chronic constipation was found in 50% (n = 10) patients, obstructive uropathy in 30% (n = 6), weight loss, chronic cough and mental illness, each in 10 % (n = 2) of patients. Primary procedure was carried out in 70% patients. Mean operative time was 178 minutes; mean length of stay was 7 days with a mean follow-up of 25 months. Early complications were noted in 5% (n = 1) patients, and late complications in 30% (n = 6). Based on this data, comparative analysis was carried out between different variables. CONCLUSIONS: Surgery is the only treatment for rectal prolapse in adults. Several procedures are done suggesting that there is no standard treatment for this ailment. The number of male patients was higher, chronic constipation was the most common risk factor, abdominal rectopexy was the favored procedure, perineal procedures were associated with shorter operative time, hospital stay, and were done more often in females and elderly. Revision surgery was associated with longer operative time and hospital stay. PMID- 16045096 TI - Unanticipated hospital admission after ambulatory surgery. AB - The unplanned admission rate is considered to be an important measure of the quality of ambulatory surgical units. The objective of our study was to evaluate the unanticipated hospital admission rate from the Surgical Day Care (SDC) unit of our university affiliated teaching hospital and to analyze the reasons for admission. A review of all unanticipated admissions over a one-year period was done. The admission rate was calculated and the reasons for admission were analysed. The overall admission rate was 4.93%. Most of the admissions were ordered by the surgeons (97%). The main reasons for admission were patient observation indicated for various reasons (72%) and patient request (18%). Eighty percent of the admitted patients had received general anaesthesia. Admissions were also related to the male gender (69%), age over 65 years (27%) and surgeries ending in the afternoon (69%). On analyzing the reasons for admission, a large number of admissions were found to be due to preventable causes. We conclude that proper selection of patients, careful scheduling of lists and education of patients and clinical professionals can help to avoid many unanticipated admissions after day care surgical procedures. PMID- 16045097 TI - Apache II score as a predictor of length of stay and outcome in our ICUs. AB - The APACHE II (acute physiology and chronic health evaluation) is used widely for predicting probability of hospital mortality and length of stay in the ICU. APACHE II forms were available to all ICU residents within 24 hours of admission, and a score was assigned to them. Based on our results the APACHE II score has reliably predicted an outcome of the least amount of length of stay (LOS) in the ICU as well as a 100% probability of being shifted out of the ICU for a score of < 10 (according to international benchmarks). This reliable scoring system can be used for predicting mortality and length of stay and therefore, resource allocation, antibiotic use and ethical decisions regarding counseling families about end of life decisions--all within 24 hours of admissions. PMID- 16045098 TI - Extra pontine myelinolysis associated with hypophosphatemia. AB - We report a unique association of extra-pontine myelinolysis with severe hypophosphatemia developing in a young lady with a prolonged febrile illness and inadequate oral intake. The late identification of severe hypophosphatemia resulted in extra-pontine myelinolysis. Gradual improvement in clinical status was noticed with phosphate replacement and good supportive care. PMID- 16045099 TI - Perioperative anaesthetic management of a patient with relapsed aplastic Anaemia. AB - Aplastic anaemia occurs due to failure of blood cell production by bone marrow. All blood cell counts are low. Untreated patients may die within one year mainly due to uncontrolled infection due to neutropenia. Anaemia and thrombocytopenia cause troublesome symptoms and necessitate frequent red cell and platelets transfusions. Surgery poses a very high risk for these patients because of the several complications that can occur in the perioperative period. In this paper we report on a patient with relapsed aplastic anaemia who was optimized preoperatively and underwent a total abdominal hysterectomy under general anaesthesia. PMID- 16045100 TI - Prescription pattern of benzodiazepines for inpatients at a tertiary care university hospital in Pakistan. AB - OBJECTIVE: To determine the point prevalence of benzodiazepine prescriptions for inpatients at a tertiary care university hospital in Pakistan and to correlate it with prescription patterns of various specialties, indications and demographic variables of the patients. METHODS: This 24-hours point prevalence study was done at The Aga Khan University Hospital, Karachi. By convenient random sampling, 208 inpatients were interviewed. Patients' files were also studied to record the drugs administered. Data was entered into questionnaires and analyzed by SPSS 10.0. RESULTS: The point prevalence of the benzodiazepines was 21.2%. It was higher among males than females and among surgical than non-surgical patients. Midazolam was the most commonly used benzodiazepine, followed by Alprazolam and Lorazepam. Pre-anesthesia and psychiatric symptoms were the two most common indications. Oral route was used in 84% patients for drug administration and mean Valium equivalent dosage was 4.86 mg/day. Mean length of prescription was 3 days. Longer duration of hospitalization was a significant predictor of the requirement of benzodiazepine prescription (p-value = 0.020). CONCLUSION: Prescription pattern of benzodiazepines at a tertiary care university hospital is similar to that reported in the developed countries through monitoring at various levels by physicians, clinical pharmacist and nursing staff. Data regarding the prescription pattern of benzodiazepines is scarce, and it needs to be expanded to formulate clear guidelines regarding their prescription. PMID- 16045101 TI - Efficacy of local omeprazoles in the treatment of peptic ulcer disease. PMID- 16045102 TI - Creating competition and accountability in a changing healthcare environment. AB - Many problems in the healthcare system mirror the structure of a competition in which the participants divide--and sometimes destroy--value instead of creating it, according to a recent study. Some healthcare leaders say that these problems can be dealt with by emphasizing accountability throughout a healthcare organization. PMID- 16045103 TI - Taking the quality challenge: highlights from the AHRQ summit. AB - When it comes to quality in healthcare delivery, much has emerged about best practices-backed up by scientific data. But the results still are not there, the director of the Agency for Healthcare Research and Quality, Carolyn Clancy, MD, said at a recent healthcare quality summit sponsored by the agency. Instead, the healthcare community has to challenge itself to do better. PMID- 16045104 TI - "Hospital compare" gets official rollout by CMS. AB - Hospital Compare Gets Official Rollout by CMS After almost 2 1/2 years of planning and discussion, the Centers for Medicare & Medicaid Services (CMS) and the Hospital Quality Alliance officially rolled out the "Hospital Compare" website last month. With it, healthcare professionals and consumers can compare parameters of healthcare quality at most hospitals. PMID- 16045106 TI - Conceptualisation of community-based basic nursing education in South Africa: a grounded theory analysis. AB - Community-based education is about a decade old in basic nursing education in South Africa. An extensive review of literature revealed that although CBE was a familiar concept in South Africa, there was however, limited understanding of what this phenomenon means. The purpose of the study was to analyse the concept 'community-based education' with the aim of discovering shared understanding of this phenomenon in basic nursing education within the South African context. Strauss and Corbin's (1990) grounded theory approach was used to guide the research process. The South African Nursing Council's (SANC) education committee, the National Department of Health, human resources division representatives as well as seven nursing education institutions with well-established CBE programmes participated in the study. The data was collected by means of observations, interviews and document analysis. Purposive sampling and later theoretical sampling was used for selecting interviewees. This resulted in a total of 45 interviewees. The data collection and initial data analysis took place concurrently. Descriptive analysis followed by conceptual analysis was performed using Strauss and Corbin's model. The findings in this study revealed that community-based education is education that uses the community extensively, especially the under-developed and under-resourced settings, for learning purposes in order to enhance relevance of nursing education to the needs of the South African population. The core discriminatory characteristics of CBE were found to include; primacy of the community as a learning environment; the early exposure of students to community-based learning experiences; community-based learning experiences dominating the curriculum, exposure to community-based learning experiences throughout the curriculum, vertical sequencing of community based learning experiences in a curriculum, starting from primary settings to secondary and later tertiary health care settings to facilitate the development of competencies required when serving in all these settings, and lastly, learning through providing service to the under-resourced communities. Community involvement and partnership, problem-centred learning, valid assessment of learning emanated as important characteristics of CBE but which were identified as gaps in the existing programmes. Recommendations focused on these gaps as well as to the problem of community-based learning experiences which were mainly concentrated in first and/or second year levels in most of the programmes. PMID- 16045107 TI - Community perception of quality of (primary) health care services in a rural area of Limpopo Province, South Africa: a qualitative study. AB - The aim of the study was to survey perceptions of quality of (primary) health care services provided in rural communities in the Limpopo province. Ten focus groups discussions were held with community members chosen by convenience from public places from four villages in the central region of the Limpopo Province. The sample included 42 women and 34 men (76 participants). Results indicated perceived quality discussed within the following categories: (1) conduct of staff (reception, communication, discrimination, care and compassion, respect for privacy), (2) technical care (examination, explanation of treatment, responsiveness, treatment outcomes), (3) health care facility, (4) health care organisation, (5) drugs (availability, explanation, effectiveness, payment), and (6) waiting time. The findings suggest some satisfaction with free basic and preventive health care and social services provided but there is a need to look closely into the interpersonal dimension of the services provided, provision of medication with adequate explanation to patients on the medication given, and on structural aspects, there is need for the government to give support to the clinics to provide adequate services. Improving drug availability, interpersonal skills (including attitudes towards patients) and technical care have been identified as the three main priorities for enhancing perceived quality of primary health care and health policy action. PMID- 16045108 TI - A model for facilitation in nursing education. AB - A key step in the development of a model for learning is to identify the needs of both the facilitators and the learners. The purpose of this study was to develop a model for facilitation in nursing education based on the identified needs of facilitators and learners and a confirmation by the literature. A qualitative research design was used and the population of the study was the facilitators and the learners of departments/schools of nursing in four universities in South Africa. Focus group interviews were conducted with facilitators and learners. The sample was purposively selected. The results underline the needs as well as the perceptions of both facilitators and learners with regard to facilitation. Concepts were identified and analyzed from the identified needs which led to the development of the model for facilitation. The complete visual model was described and presented to experts for evaluation. Feedback from the evaluators was used to adapt and finalize the model, after which the model was again presented to evaluators who approved and accepted it. The model is a unique contribution to nursing education as it sets guidelines for a new field of learning in nursing education. PMID- 16045109 TI - The meaning of community involvement in health: the perspective of primary health care communities. AB - The goal of this study was to establish the understanding and appreciation of the essence of PHC principles in the two Primary Health Care (PHC) communities. The PHC communities in this study referred to the people who were involved in the operation of the phenomenon, that is health professionals working in the health care centers and the communities served by these health care centers. It was hoped that the study would enhance the understanding of the importance of community involvement in health (CIH) in health care delivery, for both community members and health professionals. A case study method was used to conduct the study. Two community health centers in the Ethekwini health district, in Kwa Zulu Natal, were studied. One health center was urban based, the other was rural based. A sample of 31 participants participated in the study. The sample comprised of 8 registered nurses, 2 enrolled nurses, 13 community members and 8 community health workers. Data was collected using individual interviews and focus groups, and was guided by the case study protocol. The findings of the study revealed that in both communities, participants had different, albeit complementary, understanding of the term 'Community Involvement in Health' (CIH). Essentially, for these participants, CIH meant collaboration, co-operation and involvement in decision-making. PMID- 16045110 TI - Competence of newly qualified registered nurses from a nursing college. AB - The South African education and training system, through its policy of outcomes based education and training, has made competency a national priority. In compliance to this national requirement of producing competent learners, the South African Nursing Council (1999 B) require that the beginner professional nurse practitioners and midwives have the necessary knowledge, skills, attitudes and values which will enable them to render efficient professional service. The health care system also demands competent nurse practitioners to ensure quality in health care. In the light of competency being a national priority and a statutory demand, the research question that emerges is, how competent are the newly qualified registered nurses from a specific nursing college in clinical nursing education? A quantitative, non-experimental contextual design was used to evaluate the competence of newly qualified registered nurses from a specific nursing college. The study was conducted in two phases. The first phase dealt with the development of an instrument together with its manual through the conceptualisation process. The second phase focused on the evaluation of the competency of newly qualified nurses using the instrument based on the steps of the nursing process. A pilot study was conducted to test the feasibility of the items of the instrument. During the evaluation phase, a sample of twenty-six newly qualified nurses was selected by simple random sampling from a target population of thirty-six newly qualified registered nurses. However, six participants withdrew from the study. Data was collected in two general hospitals where the newly qualified registered nurses were working. Observation and questioning were used as data collection techniques in accordance with the developed instrument. Measures were taken to ensure internal validity and reliability of the results. To protect the rights of the participants, the researcher adhered to DENOSA'S (1998:2.2.1) ethical standards of research. A descriptive statistical method of data analysis was used in this study. Findings revealed that newly qualified registered nurses were not competent. The highest score obtained was 51% and the lowest score was 22% with an average score of 34.05%. The results concerning the implementation of the phases of the nursing process indicated that participants were fairly competent in terms of knowledge, skills, attitudes and values of assessment. Participants had very little knowledge of nursing diagnosis and were not competent on the skills of diagnosis. Participants lacked basic knowledge, skills, attitudes and values of the nursing process. They lacked critical thinking skills in their approach to providing quality patient care. The recommendations of the study relate to improving the system of clinical accompaniment, reviewing the clinical facilities where learners are allocated, reviewing the implementation of the curriculum, the methods of teaching and the quality assurance mechanisms that are in place. Further research is recommended on competence of newly qualified registered nurses at other nursing colleges or similar context. PMID- 16045111 TI - Aspects of the working life of women in the nursing profession in South Africa: survey results. AB - This article reports on a survey done among nurses registered with the South African Nursing Council. The survey was carried out in the last quarter of 2003. The purpose of the survey was to investigate aspects of the working life of women in the nursing profession in South Africa and to make recommendations on how their working environment could be improved. The important findings were that pay related issues dominate as the main problem at work. Improving pay scales and being paid according to extra experience, responsibilities and qualifications could improve the nurses' working environment. Furthermore, training opportunities, medical insurance and equal opportunities should be addressed as a matter of urgency. In general, respondents had a positive attitude towards their job, which leaves the impression that nurses still regard their jobs as something they do for the sake of a service to the community and not only for the money they earn. PMID- 16045112 TI - Contributing to the ICNP: validating the term cultural diversity. AB - AIM: The specific aims of this study were to: Propose a definition of the term cultural diversity; Validate the term cultural diversity; and Submit a term and definition for international utilisation to the International Council of Nurses (ICN) for consideration for inclusion in the ICNP. BACKGROUND: South Africa was one of four African countries (Botswana, South Africa, Swaziland, and Zimbabwe) funded by the WK Kellogg Foundation to participate in the ICNP project. South Africa had 2 research groups. One of the research groups identified the term cultural diversity to define. METHOD: This was a qualitative study where a philosophical perspective was used to explore, explain and describe nursing practice. The combined method proposed by the International Council of Nurses (ICN) was utilised to define and validate the term cultural diversity. FINDINGS: Validation and literature review provided sufficient support for the defined characteristics and the term was finally defined and submitted to ICN in November 2002 as: CULTURAL DIVERSITY is a type of CULTURE with the specific characteristics: co-existence of different groups, e.g. ethnic, religious, linguistic and other groups each with their own values and belief systems, traditions and different lifestyles. CONCLUSION: The research group was informed in December 2003 of the ICNP Evaluation Committee recommendation that the term cultural diversity will be included in the ICNP. PMID- 16045113 TI - The effect of curricula approaches to the development of the student's clinical reasoning ability. AB - A comparative analysis was undertaken using descriptive and cross sectional survey design, to explore the effect of Problem Based Learning and Traditional approach on the development of clinical reasoning abilities of nursing students. Using quota sampling, a sample of 87 subjects was used from two University Nursing Departments, each using these curricula approaches. Students from their first, second and fourth year were interviewed, using the Triple Jump Exercise as an instrument to collect data. Data analysis using the SAS computer software package was employed to obtain both descriptive and statistical summarizations. Though descriptive analysis of the scores of clinical reasoning showed a slight difference between the two curricular approaches, this was not confirmed statistically as the two factor ANOVA and Tukey's methods revealed no significant differences by approaches. The only significant difference was revealed between the students' levels of study with senior levels (4th years) outperforming their juniors. These findings therefore conclude that, students using Problem Based Learning and Traditional approach perform on a similar level in clinical reasoning. PMID- 16045114 TI - [Modern trends in liver surgery]. PMID- 16045115 TI - [Minimally invasive saphenous vein harvesting for coronary artery bypass grafting. Use of Vega system]. AB - The authors present their first experience with minimally invasive saphenous vein harvesting for coronary artery bypass grafting (CABG). From February 2004 to June 2004 33 patients undergoing CABG were scheduled for minimally vein harvest with the use of Vega system (B/Braun-Aesculap, Tuttlingen, Germany). We evaluated wound and neurological complications associated with the vein harvest at the time of discharge and at the 3-month follow-up. The limited experience suggests that minimally invasive vein harvesting offers improved patient outcome in term of wound healing. PMID- 16045116 TI - Intraoperative frozen section examination of the sentinel lymph node in breast cancer. AB - AIM OF THE STUDY: Intraoperative frozen section (FS) examination of the Sentinel node (SN) in breast cancer patients is questioned due to the relatively high number of positive SN(s) found in the permanent histological examination. This study reviews the data of the Austrian sentinel node study group on FS examination of the SN and tries to identify patients with a high risk of incorrect negative results. METHODS: 2326 breast cancer patients of the Austrian Sentinel node study group who underwent SN biopsy and intraoperative FS examination of the SN were further analysed for incorrect negative results and clinicopathologic factors indicating a higher rate of incorrect negative results. RESULTS: The FS of the SN was positive in 513 of 2326 patients (22.1%) and negative in 1813 of 2326 patients (77.9%). Permanent histological examination revealed a metastatic SN in 282 of 1813 patients. (incorrect negative rate 15.6%). 158 of 282 patients (56%) were found through H&E serial sectioning, whereas 124 of 282 patients (44%) were only seen in immunohistochemistry. Micrometastases, lobular histology and preoperative chemotherapy were associated with a higher rate of incorrect negative results. CONCLUSION: Incorrect negative results of FS examination are seen in 15% of patients and require a secondary axillary lymph node dissection. The disadvantage of missing a positive SN through FS is by far outweighed by the advantage of a single stage operation in case of a positive SN. PMID- 16045117 TI - [Experimental use of the peritoneal graft as a vascular prosthesis]. AB - INTRODUCTION: The parietal peritoneum appears to be a suitable material for the vascular system reconstructions. AIM: The aim was to assess and compare thrombogenicity and ability for endothelization of the mesothelial and submesothelial side of the parietal peritoneum in the canine venous system. MATERIAL AND METHODOLOGY: EXPERIMENTAL ANIMALS: canis familiaris (n = 13), half breeds of both sexes, aged between 1 and 2 years, weighting 15-25kgs, underwent authological transplantations of the peritoneal grafts with the mesothelial side in the lumen- the group M (n = 5) and with the submesothelial side in the lumen the group S (n = 5). In the control group K (n = 3) a part of the venous wall was used as a graft and was affixed back to its original place. The bioptic samples collected on the 10th, 20th, 30th and 40th postoperative day (POD) were stained using the HE staining, NADPH-d and imunohistochemically on the intermedial filaments. The endothelization rate of the peritoneal graft was measured using morphometry and the trombogenicity was assessed peroperatively. RESULTS: In none of the trial groups a presence of thrombi was detected peroperatively. In the first trial group (group M), the onset of the peritoneal graft epithelization (reaching 20%) was recorded on the 10th POD. The endothelization process was completed on the 30th POD in this trial group. In the second trial group (group S), the peritoneal graft epithelization reaching 10% was recorded on the 10th POD. The process was completed on the 40th POD. In the third trial group K, no endothelial changes were recorded during the experiment. CONCLUSION: Both sides of the peritoneum do not show signs of thrombogenicity and possess ability for endothelization. PMID- 16045118 TI - [Elective standard appendectomy versus elective laparoscopic appendectomy in women. A retrospective study]. AB - AIM: Comparison of open appendectomy and laparoscopic appendectomy with focus on the wound infectious complications. METHODS: Retrospective comparison of the early postoperative complications of the elective open and elective laparoscopic appendectomy in the Surgical Clinics of the Teaching Hospital Hradec Kralove in period January 2001-July 2004. RESULTS: 250 elective appendectomies in women were performed in followed period. 187 (75%) were done by laparoscopic approach, 63 (25%) by open procedure. Conversion was necessary in 18 cases (9.6%). The average hospital stay was statistically significantly shorter following laparoscopic than open appendectomy, 4.6 +/- 1.5 days counter to 6.1 +/- 1.6. The global morbidity was comparable in both groups 15%, one patient died (0.5%) in laparoscopic appendectomy group. The incidence of early postoperative complications was not statistically different in both groups. CONCLUSIONS: The average hospital stay was statistically significantly shorter following laparoscopic appendectomy. Laparoscopic appendectomy did not improve the results regarding infectious complications and was a cause of serious complications. Some of them required operational revision. PMID- 16045119 TI - The pedicled rectus femoris muscle flap for reconstruction of complicated abdominal wall defects. AB - AIM OF THE STUDY: Large or complicated abdominal wall defects caused by recurrent incisional hernias, infections or tumor resections often require the use of prosthetic mesh, local tissue transposition or even distant muscle flaps for proper reconstruction. Due to the sometimes discouraging results of meshes muscle flaps are an appreciated alternative. We followed up a series of cases to assess the value of the pedicled rectus femoris muscle flap. METHODS: Follow up time ranged from 6 months to 4 years. 12 cases of reconstruction of the abdominal wall using pedicled rectus femoris muscle flaps after tumor resections, recurrent incisional hernias, and infection are presented. Abdominal wall stability was examined clinically. The aesthetic and the functional results were assessed using a standardized questionnaire. The loss of torque in the quadriceps muscle was evaluated using a dynamometer. RESULTS: In all but one patients a stable abdominal wall could be reconstructed. We saw no major complications. The loss of true muscular capacity in the quadriceps muscle of the operated leg was 19% compared to the nonoperated leg, but was tolerated very well. CONCLUSION: The donor site morbidity is moderate. The flap provides an easy and save possibility to reconstruct the abdominal wall. The rectus femoris muscle flap should be considered as an alternative for abdominal wall reconstruction. PMID- 16045120 TI - [Postcatheterization pseudoaneurysm of the radial artery]. AB - A postcatheterization pseudoaneurysm of the radial artery remains a rare complication, considering frequencies of its punction. The radial artery is easily accessible for the punction site management after the catheter removal. We present pathogenesis of their origin, their prevention and options for surgical management of the radial artery pseudoaneurysms. PMID- 16045121 TI - [Miniinvasive laparoscopic or retroperitoneoscopic radical nephrectomy for the parenchymal tumor]. AB - INTRODUCTION: Laparoscopic (LRNE) or retroperitoneoscopic (RRNE) radical nefrectomy originally published in 1991, respectively in 1993, began to be introduced in common practice at the end of the 20th century. In this work we summarize our findings gathered upon introduction of these methods. SUBJECTS AND METHODOLOGY: We present a group of 50 miniinvasive nephrectomies conducted during 2003-2004 (46 LRNE and 4 RRNE). LRNE was combined with ipsilateral adrenalectomy in five cases and with cholecystectomy in three cases (in all cases the LRNE was on the right side). RESULTS: The mean age of the patient subjects was 65 +/- 11 years (34-81). The blood loss was 134 +/- 201 ml (30-1200). The tumor size according to the CT was 50 +/- 13 mm (30-82). Histologically, 45x renal carcinomas, 4x oncocytomas, 1 x urothelial carcinomas were diagnosed. The drain was removed 2.1 +/- 0.9 days following the surgery. The mean hospitalization time was 6.1 +/- 1.6 days (4-9). Only a single major, however lethal, complication arised: In a 74-year old female, the left-sided LRNE due to the carcinoma pT3bG2 and at the same time vaginal hysterectomy, extraction of the renal preparation through the vagina and anterior and posterior vaginoplasty (for a prolaps) were conducted. The patient was repeatedly revised for haemoperitoneum and she exited on the 18th postoperative day due to pneumonia. The mean duration of the LRNE was 168 +/- 40 minutes (80-265). The combined LRNE and cholecystomic procedure lasted 265 minutes. The mean duration of the LRNE with CHE was 213 minutes, and of the LRNE with adrenalectomy was 170 minutes. In 4 cases of the RRNE (indicated three times due to preceeding major surgical procedures in the abdominal cavity and once for the Crohn's disease) the mean duration was 203 minutes. CONCLUSION: Miniinvasive RNE in TI tumors is a modem reproducible methodology suitable for application in clinical practice. The laparoscopic approach appears more appropriate. The retroperitoneoscopic approach is indicated mainly in cases of postoperative adhesions in the abdominal cavity. PMID- 16045122 TI - [Measurement of carpal tunnel pressure during surgery of carpal tunnel syndrome]. AB - A significance of carpal tunnel pressure measurement during surgery of carpal tunnel syndrome (endoscopic and twin incision technique) was evaluated. The pressure in different levels of palm and in different positions of the hand was measured by ICP sensor Codman during operation of 10 patient. The results are presented in the form of several graphs. In all 10 cases excellent results were obtained. PMID- 16045124 TI - Colorectal cancer: lifestyle and dietary factors. AB - INTRODUCTION: Colorectal cancer is the most common tumor in the developed countries, and the number of new cases annualy is aproximately equal for men and women. Several environmental factors can interact in all steps of carcinogenesis. Lately the balance between genetic predisposition and these factors, including nutritional components and lifestyle behaviors, determines individual susceptibility to develop colorectal cancer. The aim of this study is to revise the references about lifestyle include diet, physical exercise, tobacco smoking and use of alcohol, and the risk of colorectal cancer in databases published during 1994-2004. DIETARY FACTORS: According to the reports high intake of red meat, and particularly of processed meat and positive energetic balance (high intake of total fat and carbohydrate) was associated with a moderate but significant increase in colorectal cancer risk. Convincing preventive factors include increase consumption of a wide variety of fruit and vegetable, particularly, dark-green leafy, cruciferous, a deep-yellow on tones, and fibre. LIFESTYLE: Physical activity as a means for the primary prevention of colorectal cancer. There is a probable synergic effect among physical inactivity, high energy intake and obesity and incidence of colorectal cancer. A growing body of evidence supports that avoidance overweight and the use of tobacco and alcohol is recommended to prevent colorectal cancer. CONCLUSION: Current data suggest that lifestyle modification including proper diet such as the ones rich in vegetable and poor in red meat and fat, regular physical activity and maintaining an appropriate body weight and avoiding the use of tobacco and alcohol may lead to reduce colorectal cancer risk. PMID- 16045125 TI - [Home parenteral nutrition, question mark in pediatrics]. AB - Home parenteral nutrition (HPN) is a necessary but under-used therapy in Spain as compared to other European countries. This problem has been studied at the 1st Discussion Forum of the Spanish Society for Parenteral and Enteral Nutrition. One of its recommendations has been "to design studies in order to analyze the problems detected in HPN practice". Our study attempts to assess pediatric professionals knowledge and opinion about this concern, because children are important recipients of this therapy. A multidisciplinary team prepared a questionnaire to be distributed among 136 health professionals (70 physicians and 66 nurses) working in 4 public hospitals (H) and 8 Primary Health Care Centers (PHC) from Asturias and Salamanca (Spain). Answers show that few professionals have appropriated theoretical and practical knowledge about HPN. Most of them have received neither academic training education nor information over the last year on this matter. The ability to solve patient and caregiver doubts in daily management (handling, storage, compatibility and nutrition stability, parenteral infusion equipment problems, etc) is only 10% in PHC and 50% in H. Only 6 physicians (4.4%) from H group know about the existence of NADYA expert group despite its website and handbooks providing a wide information about HPN. The advantage of HPN to improve life quality, its economic efficacy and the need of cooperation to this respect between H and PHC are matters with a high level of agreement. An important number of pediatric professionals does not have enough knowledge about HPN in PHC. The awareness of HPN must be promoted among clinical services dealing with patients able to benefit from this therapy. All available resources must be used to improved this issue. Specific protocols developed by multidisciplinary teams would promote H and PHC collaboration to help patients, who are the aim of our activity. PMID- 16045126 TI - [The year 2002 national register on home-based parenteral nutrition]. AB - AIM: To report on the results of the Registry on Home-based Parenteral Nutrition (HPN) of the NADYA-SENPE working group, corresponding to the year 2002. MATERIALS AND METHOD: Compilation of the registry data loaded by the Units in charge of HPN patients care. It consists of an on-line registry available to the registered users of the group's web page (www.nadya-senpe.com). Epidemiological, diagnostic, access route, complications, hospital admissions, degree of disability, and course until December 31st of 2002. RESULTS: Data from 74 patients were gathered (56.8% women and 43.2% men), from 18 hospital centers. Mean age of adult patients was 49.4 +/- 15.5 years and 2.3-1.1 years for patients younger than 14 years (n=3 patients). Diseases that prompted the use of HPN were mesenteric ischemia (29.7%), followed by neoplasms (16.2%), radiation enteritis (12.2%), motility impairments (8.1%), and Crohn's disease (5.4%). Tunneled catheters were used in 52.7% of cases, as compared to 36.5% of subcutaneous reservoirs. Mean treatment duration was 8.7 +/- 4.4 months; 68.9% of patients remained on HPN for a duration longer than 6 months, and in 41.9% longer than one year. Patients' follow-up was mainly done from the reference hospital (87.8%), and the remaining patients (12.5%) by the home care team. In no case patients were followed by the primary care team or other specialists than the ones that prescribed nutritional support. In 94 cases there were complications related to nutritional therapy. The more frequent complications presented were infectious. These complications represented 1.84 admissions per patient. The mean number of visits was 12.9 per patient (10.2 routinary visits and 2.7 emergency visits). At the end of the year, we observed that 74.3% patients stayed in the program, whereas in the remaining 23.6% HPN had been discontinued. The main causes for discontinuation were death (52.9%), and switch to oral diet (23.5%) or enteral nutrition (11.8%). With regards to disability degree, 16.1% were confined to a wheelchair or bed, and 17.6% had no disability at all or only a mild social disability. CONCLUSIONS: We observed a sustained HPN prevalence rate in Spain (1.8 patient pmp). The main cause for its use was short bowel syndrome secondary to vascular disease, followed by cancer. Complications associated to nutritional therapy were common, especially of infectious origin. PMID- 16045127 TI - [The year 2002 national registry on home-based enteral nutrition]. AB - GOAL: To communicate the information available by the NADYA-SENPE Working Group from patients on Home Enteral Nutrition (HEN) in our country during the year 2002. MATERIAL AND METHODS: The data were collected through a closed questionnaire included in the web site of the Working Group (www.nadya-senpe.com) available only by the authorized users. Variable included were: epidemiological information, the indication to prescribe this treatment, the access path, the specific nutritional formula used, the treatment duration, the complications and hospital readmission related to the nutritional treatment, the follow-up and the quality of life. RESULTS: We register 3967 patients that belong to twenty-one hospitals. Mean age from those adults 69.2 +/- 19.2 years, and from those younger than 14, 5.6 +/- 4.1 years. Neurological and neoplasic diseases were the diagnostics more frequents (39.2% and 34.6%, respectively). Oral nutrition was the preferential rout used for the enteral nutrition (53.6%) followed by naso enteral tube (30.6%), and only in 15.8% we used ostomy tubes. Polymeric was the enteral formula mainly utilized (81.5%). The mean time on HEN was 5.8 +/- 4.4 months; the 35.7% of patients stayed in the treatment for less than 3 months, 22.4% between 3 and 6 months, and 41.6% more than 6 months. Patients were followed mainly by Nutritional Support Unit from the reference hospital (75.3%). While the reference hospital supplies the material (65.7%), reference hospital pharmacy (43%) and public pharmacies (37.3%) provides the enteral formula. Complications related to enteral nutrition included change of enteral tube (29.7%), mechanical complications (22.9%), gastrointestinal complications (22.9%), and the metabolic one (9.2%). These complications were followed by 0.02 hospitalizations/patient. At the end of the year, 49.3% of patients were in the HEN programme, and in 41.5% HEN was finish due to accept oral conventional alimentation (47.3%) or by deceased of patients. While 31.8% of the patients were confined to bed o armchair, 17.8% no o light discapacity degree was observed. CONCLUSIONS: We found a persistence of these treatment in our country (96.5 patients/million inhabitants. Neurological and neoplasic diseases were the more frequent diagnosis in patients analysed. The high prevalence of cancer patients could be the main cause of oral access for enteral nutrition. Change of enteral tube was the more frequent complication observed during this treatment. PMID- 16045128 TI - [Are the tools recommended by ASPEN and ESPEN comparable for assessing the nutritional status?]. AB - INTRODUCTION: [corrected] There is no "gold standard" for identification of malnutrition. The ASPEN board of directors (2002) suggest the subjective global assessment (SGA) and ESPEN (2002) recommend the nutritional risk screening-2002 (NRS-2002) to detect the prevalence of malnutrition. AIMS: This cross-sectional study aims (1) to assess the prevalence of malnutrition on admission and (2) to know association between two tools used to evaluate nutritional risk. MATERIAL AND METHODS: 135 patients (42.2% women and 58.8% men, 62.1 +/- 14.4 years) are studied at admission. Different parameters are assessed to evaluate nutritional state. Patient's energy requirements (Harris Benedict x Long's factor) and energy intake, calculated as 24 h-recall, are compared. The prevalence of malnutrition is assessed with two screening tools: SGA and NRS-2002. RESULTS: 42.2% of the patients had lost more than 5% body weight and 39.3% had an intake lower than required at admission. The prevalence of malnutrition is 40.7 and 45.1/100 patients admitted at hospital, if SGA or NRS-2002 are used, respectivility. There is strong agreement between results of two nutritional assessment methods (p = 0.000). Serum albumin and protein concentrations and linfocytes count are less in the malnourished patients. CONCLUSIONS: The prevalence of malnutrition is elevated. In clinical practice, both methods could be used to identify patients at nutritional risk, but the NRS-2002 is a less subjective method. PMID- 16045129 TI - [Multicenter study on incidence of total parenteral nutrition complications in the critically-ill patient. ICOMEP study. Part I]. AB - OBJECTIVE: To assess parenteral nutrition complications in a prospective cohort of patients admitted to the ICU. MATERIAL AND METHODS: Prospective, multicenter study of patients admitted to the ICU and that received total parenteral nutrition (TPN). A 14-item questionnaire was done. Total number of admissions and TPN-treated patients were recorded. Demographical data, diagnosis, APACHE II, multiorgan dysfunction index and TPN indications were included. Each complication was previously defined and was related to the route, administration, serum electrolytes unbalances, active ingredients, liver dysfunction, and nosocomial infections. An independent group managed the databases. Data were expressed in absolute values or by their median and percentile (25-75). The rate and incidence density of complications are determined. RESULTS: The number of admissions during the study period was 3409. Three hundred and seventy (11%) were valid cases that were treated with PN. Two hundred and thirty seven were men, 142 patients were admitted for medical causes, 195 for surgical causes and 33 for trauma. One hundred and twenty patients were moderately or severely underfed, 181 received PN for 7 days (3-11), and 189 received PN and EN. The main indication for PN was paralytic ileus, in 145 patients. TPN was withdrawn for switching to EN in 121 cases and for complications in 5 cases. PN summed up 3220 days. The incidence of complications was: access route 9.32%, maintenance 19.7% and 0.8 per 100 days. Administration, 5.6% and 6.4 per 100 days. Nutrients, 94.3% and 10.8 per 100 days. Catheter-induced sepsis, 54.6% and 0.44 bacteriemias per 100 days with catheter. Nosocomial infection, 54.6% and 6.27 per 100 days on TPN. CONCLUSIONS: hyperglycemia and liver dysfunction have a greater incidence. Hyperglycemia, electrolyte unbalances and catheter complications have a greater number of therapeutic interventions, but does not mean that TPN should be withdrawn. PMID- 16045130 TI - [Multicenter study on incidence of total parenteral nutrition complications in the critically-ill patient. ICOMEP study. Part II]. AB - OBJECTIVE: To assess enteral nutrition complications in a prospective cohort of patients admitted to the ICU. MATERIAL AND METHODS: Prospective, multicenter study of patients admitted to the ICU and that received enteral nutrition (EN). Demographical data, main diagnosis, and nutritional and severity indexes were included. Complications were previously defined and were categorized as gastrointestinal or infectious. An independent group managed the databases and performed the statistical analysis. Data were expressed in absolute values or by their median and percentile (25-75). The rate and incidence density of complications and the odds ratio for complications were determined. RESULTS: 544 out of 3409 patients received enteral nutrition. Three hundred and three patients were admitted for medical causes, 149 for surgical causes and 92 for trauma. Ninety nine patients were underfed, and the nutritional risk index was 69.7. The incidence of gastrointestinal complications was 6 episodes per 100 days of nutrition, and in 89 patient they resulted in enteral nutrition withdrawal. Early enteral nutrition did not yield more complications than late EN. Vomiting was more frequent in late EN (OR 0.4; 95%CI: 0.2-0.7). Temporary withdrawal of diet in 140 patients was the most used procedure, which resulted in a lower caloric intake (median of 2 kcal/kg/d) than programmed. Nosocomial infection rates were 7.47% for catheter-induced sepsis and 51% for other nosocomial infections, and 35% developed pneumonia. CONCLUSIONS: Gastrointestinal complications with enteral nutrition are frequent, result in insufficient caloric intake and definitive withdrawal of diet in a significant number of cases. Early enteral nutrition is not associated with an increased number of complications. Episodes of increase of the gastric residue do not increase the incidence of pneumonia. Other infectious complications are within the range of what has been published and seem to be little modified by the use of enteral nutrition. PMID- 16045131 TI - [Nutritional status of an hospitalised aged population]. AB - AIMS: The aims of the study were to evaluate the nutritional status of the hospitalised elderly patients, to estimate the prevalence and distribution of malnutrition among them and to value the correlation of the Mini Nutritional Assessment (MNA) with nutritional biochemical and anthropometric parameters. MATERIAL AND METHODS: A cross sectional survey was done among a sample of 200 elderly patients admitted to the USP San Carlos Hospital (Murcia) during a period of 3 months. The study employed the MNA and a blood test including hemogram and albumin and transferrin serum levels. Chang's protocol was used to classify the nourishing pattern. For statistical analyses of the MNA tool, the Spearman's coefficient was employed. RESULT: Mean age of the 200 patients was 80.72 SD 7.43 years. Mean weight was 63.41 SD 19.57 kg and mean height was 160.93 SD 8.36 cm, with a mean Body Mass Index of 24.27 SD 7.31. The serum values of albumin and transferrin and total lymphocytes in blood were 3.09 SD 0.5 g/l, 1.69 SD 0.37 mg/dl and 1412 mm3 respectively. Average score of the MNA was 15.9 SD 6.21 with a maximum value of 27 and a minimum of 5.5 points. According to this scale, 50% (n=100) of the patients were malnourished, and 32% (n=32) of them presented mixed undernutrition of moderate degree. Values of malnutrition in the MNA test correlated significantly (p < 0.001) with values under normality of the biochemical and immunological parameters. CONCLUSIONS: Patients over 80 years of age are either malnourished or at risk of malnutrition. These results are obtained both by means of the nutritional classic valuation and the MNA and presents a statistically significantly correlation. Therefore the MNA represents a useful, reliable and easily administered clinical tool to identify elderly people at risk of malnutrition and guide nutritional intervention in order to improve nutritional status. PMID- 16045132 TI - [Cholesterol oxidation products in fresh and frozen shrimps, raw and grilled]. AB - Cholesterol oxidation products (COPs) have been related to different toxic effects, being the atherosclerotic process one of the best known. The presence of cholesterol oxides in freshly and frozenly commercialised shrimps, both raw and grilled, was studied. The determination was made by gas chromatography-mass spectrometry (GCMS). Fresh shrimps showed significant amounts of all analysed COPs, except for 7alpha-hydroxycholesterol, accounting in total for 33.15 microg COPs/g fat. In contrast, in frozen commercialised shrimps only 7-ketocholesterol and 7beta-hydroxycholesterol were detected. These results point out the great effectiveness of the commercialisation of this type of products under freezing, in terms of to the minimisation of the COPs formation. The cooking method (grilling) increased the COPs content in both types of shrimps, reaching 55.43 microg COPs/g fat in fresh shrimps and only 13.06 microg COPs/g fat in frozen ones. PMID- 16045133 TI - [The skeleton in the hospital closet. 1974]. PMID- 16045134 TI - [Bilateral corneal ulceration as a result of caloric-protein malnutrition and vitamin A deficit in a patient with chronic alcoholism, chronic pancreatitis and cholecystostomy]. AB - Since the discovery of vitamins, there has been an increasing interest at relating vitamins with particular diseases. In particular, for vitamin A its singular importance has been determined in multiple vital functions, and its relationship with diseases, both in deficit and in excess, is nowadays completely demonstrated. In developed countries, vitamin deficiency-related diseases have been greatly reduced; however, in some patients with particular features they must be kept in mind. This is the case of a 45 year-old man, with a history of chronic alcoholism, non insulin-dependent diabetes meIlitus and cholecystectomy with a high biliary drainage secondary to emphysematous cholecystitis and perivesicular abscess. He complains of bilateral ocular pain, photophobia, and decreased visual acuity besides a history of pasty, sticky and foul-smelling feces. He is admitted in the Ophthalmology Department and bilateral corneal ulceration is diagnosed. A consultation to the Nutrition Department is made because of cachexia. Severe caloric and mil protein hyponutrition is observed with a BMI of 18.2 and a 23% weight loss for the last 6 months, fat-soluble vitamins (A, D and E) deficit, mild fat malabsorption, and macrocytic and hypochromic anemia. The patient's diet is supplemented with a special hyperproteinic and hypercaloric diet for diabetics, deficient vitamins and pancreatic enzymes to improve absorption are administered, and glycemia is controlled with insulin. Four months later, the patient is assessed and has a BMI of 20, anemia has resolved and from an ophthalmologic viewpoint the course is favorable, the ulcers improve and visual acuity is almost completely recovered. In chronic alcoholic patients with a low dietary intake and clinical complications with nutritional repercussions (pancreatitis that produces malabsorption or cholecystectomy with biliary percutaneous drainage) we should not forget that micronutrients deficits may explain the etiology of other associated diseases, in the present case corneal ulceration. PMID- 16045135 TI - Opioids, transporters and the blood--brain barrier. PMID- 16045136 TI - The impact of the type of anaesthesia on cognitive status and delirium during the first postoperative days in elderly patients. AB - BACKGROUND AND OBJECTIVES: Postoperative confusion and delirium is a common complication in the elderly with a poorly understood pathophysiology. The aim of this study was to examine whether the type of anaesthesia (general or regional) plays a role in the development of cognitive impairment in elderly patients during the immediate postoperative period. METHODS: Forty-seven patients > 60 yr of age and undergoing major surgery were randomly allocated to receive either regional or general anaesthesia. The mental status of the patients was assessed preoperatively and during the first three postoperative days with the Mini Mental State Examination. The incidence of delirium was also examined during the same period with the use of DSM III criteria. RESULTS: Overall, during the first three postoperative days, the mean Mini Mental State Examination score decreased significantly (P < 0.001). However, this decline was very significant only in patients assigned to receive general anaesthesia (P < 0.001) compared to regional anaesthesia. Nine patients developed delirium but the type of anaesthesia did not affect its incidence. The only important factor for the development of delirium was preexisting cardiovascular disease irrespective of anaesthesia type (P < 0.025). CONCLUSIONS: Elderly patients subjected to general anaesthesia displayed more frequent cognitive impairment during the immediate postoperative period in comparison to those who received a regional technique. PMID- 16045137 TI - A comparison of lidocaine 2% with levobupivacaine 0.75% for sub-Tenon's block. AB - BACKGROUND AND OBJECTIVE: To compare the onset of action, and quality of block, of lidocaine 2% with levobupivacaine 0.75% for sub-Tenon's block in patients undergoing cataract surgery. METHODS: We performed a two-centre trial in 91 patients who were randomized to receive 4 mL of lidocaine 2% (n = 44) or levobupivacaine 0.75% (n = 47) for sub-Tenon's block, both with hyaluronidase 15 IUmL(-1). Onset of akinesia was assessed every 2 min for 10 min. Numbers of patients requiring supplementary injections to achieve clinically satisfactory akinesia or rescue analgesia were recorded. Data were analyzed with Fisher's exact test, U-test and t-test where appropriate. Results were considered significant when P < 0.05. RESULTS: The speed of onset was statistically significantly faster for lidocaine compared to levobupivacaine (3.02 vs. 5.06 min, P < 0.001). There was no statistical difference in number of patients requiring a supplementary injection of local anaesthetic (levobupivacaine 3 vs. lidocaine 0, P = 0.24), rescue analgesia with topical tetracaine (levobupivacaine 0 vs. lidocaine 2, P = 0.5), or ocular akinesia scores at the completion of surgery (lidocaine 1.4 vs. levobupivacaine 1.6, P = 0.12). Pain scores measured by a verbal analogue scale were not significantly different for injection, perioperatively or postoperatively. CONCLUSIONS: Both agents produce a rapid onset of anaesthesia when used for sub-Tenon's block. The difference between the two agents, although statistically significant, is not clinically important. PMID- 16045138 TI - Postoperative nausea and vomiting in patients undergoing total abdominal hysterectomy under spinal anaesthesia: a randomized study of ondansetron prophylaxis. AB - BACKGROUND AND OBJECTIVE: Patients undergoing total abdominal hysterectomy under general anaesthesia have a high risk of developing postoperative nausea and vomiting (PONV). The aim of this study was to evaluate the incidence of PONV in patients undergoing total abdominal hysterectomy under spinal anaesthesia with intravenous patient-controlled analgesia (PCA) using morphine and to compare its incidence with and without antiemetic prophylaxis. METHODS: Thirty-four patients undergoing total abdominal hysterectomy under spinal anaesthesia with i.v. PCA morphine postoperatively were divided into two groups. The first (n = 17) received ondansetron prophylaxis near the end of surgery while the second (n = 17) received no prophylaxis. Morphine consumption, emetic episodes (on a 3-point scale), patient satisfaction (visual analogue score), sedation and pruritus were evaluated 2, 4, 6, 9, 12, 18 and 24h postoperatively. RESULTS: Patient characteristics, postoperative morphine consumption (43.3 +/- 7.6 vs. 40.3 +/- 12.3 mg) and peristaltic recovery time (16.9 +/- 5 vs. 18.4 +/- 5.2 h) were similar in both groups. Overall nausea and vomiting were significantly lower in the ondansetron prophylaxis group than in the group without prophylaxis (52.9% vs. 88.2%, P < 0.05). Though nausea alone was higher in the prophylaxis group (41.2% vs. 29.4%), nausea with vomiting was significantly lower in the prophylaxis group (11.8% vs. 58.8%, P < 0.01). Patients' satisfaction scores were higher in the ondansetron group at all times and the difference was significant (P < 0.05) 4 h postoperatively. CONCLUSIONS: The incidence of PONV in patients undergoing total abdominal hysterectomy under spinal anaesthesia with i.v. PCA morphine is very high (88.2%). Antiemetic prophylaxis with ondansetron is highly recommended in this patients group resulting in a lower incidence of nausea and vomiting, and significantly improves patient' satisfaction and life quality in the early postoperative period. PMID- 16045139 TI - Airway management of patients undergoing oral cancer surgery: a retrospective study. AB - OBJECTIVE: This retrospective study aims to describe the airway management and benefits of nasotracheal intubation over tracheostomy in 260 patients with oral cancer undergoing surgery. METHODS AND RESULTS: The medical records of 260 patients undergoing surgery for oral cancer were reviewed for airway management during the perioperative period. Eighteen patients had previous surgery for oral cancer and were scheduled for flap reconstruction, recurrence or other complications. In 28 cases neck movement was restricted and decreased mouth opening was found in 50% of all patients because of a large growth or fixation of tissues of head and neck, oral cavity, pharynx or larynx by tumour, or radiation fibrosis. In 53 patients intubation was undertaken under spontaneous ventilation. In 20 cases the trachea was extubated in the immediate postoperative period. In 220 cases patients were extubated next morning in the intensive care unit. In none of the cases was elective tracheostomy under local anaesthesia performed before surgery for the maintenance of the airway for anaesthesia. Elective tracheostomies were done in 17 cases. Three patients remained intubated for 24-48 h because of a high suspicion of airway obstruction following extubation due to a large pectoralis major flap. These three patients received a tracheostomy because of increased oropharyngeal and laryngeal oedema. In three cases emergency tracheostomies were performed due to upper airway obstruction after extubation and in one case prolonged elective ventilation was required due to severe chest infection. CONCLUSION: Oral cancer patients have a potentially difficult airway but, if managed properly during perioperative period, morbidity and mortality can be reduced or avoided. Oral cancer patients can be managed safely without the routine use of a tracheostomy. Nasotracheal intubation is a safe alternative to tracheostomy in oral cancer patients except in some selected patients. PMID- 16045140 TI - Repeat autoclaving does not remove protein deposits from the classic laryngeal mask airway. AB - INTRODUCTION: We tested the hypothesis that repeated autoclaving removes protein deposits from the classic laryngeal mask airway (LMA). METHODS: Twenty previously used LMAs were hand washed, machine washed, dried, autoclaved and randomly allocated into four equal-sized groups for repeat autoclaving on 0 (control), 1, 2 and 3 occasions. After the final autoclave cycle, the LMAs were immersed in a protein-staining solution, rinsed, dried and a high-resolution digital image taken of the dorsal and ventral surfaces. The severity of staining was scored by two blinded observers. RESULTS: All LMAs were stained. There was no reduction in staining with repeat autoclaving. CONCLUSIONS: Repeat autoclaving does not remove protein deposits from the LMA. PMID- 16045141 TI - The persisting analgesic effect of low-dose intravenous ketamine after spinal anaesthesia for caesarean section. AB - BACKGROUND AND OBJECTIVES: To compare the analgesic effects of intrathecal fentanyl and low-dose intravenous ketamine as adjuvants to intrathecal bupivacaine for Caesarean section. METHODS: Ninety elective Caesarean section patients were randomized into three groups. Spinal anaesthesia was performed with 15 mg hyperbaric bupivacaine in all groups. Ketamine (0.15 mg kg(-1)) or an equal volume of normal saline was given intravenously immediately after initiating spinal anaesthesia in the ketamine and control group, respectively. In the fentanyl group, 10 microg fentanyl was added to the intrathecal bupivacaine. Arterial pressures, heart rate values, adverse effects, the time of first request for postoperative analgesia, visual analogue pain scores, total analgesic consumptions at 24 and 48 h were recorded in all patients. RESULTS: The time to first request for analgesia was significantly longer in the ketamine (197 min) and fentanyl (165 min) groups compared to the control group (144 min). Postoperative pain scores were significantly lower in the ketamine group than in both other groups. Although the analgesic requirements during first 24 h were significantly lower in the ketamine group, there was no significant difference between the groups during the following 24 h. CONCLUSION: Intravenous low-dose ketamine combined with intrathecal bupivacaine for Caesarean section provides longer postoperative analgesia and lower postoperative analgesic consumption than bupivacaine alone suggesting a pre-emptive effect. PMID- 16045142 TI - Effects of thoracic epidural analgesia on glucose homeostasis after cardiac surgery in patients with and without diabetes mellitus. AB - BACKGROUND AND OBJECTIVE: Even moderate hyperglycaemia increases mortality/morbidity after coronary artery bypass grafting, stroke and myocardial infarction. The goal of this prospective study was to determine if using thoracic epidural analgesia from start of surgery until the end of the third postoperative day would blunt postoperative hyperglycaemia. METHODS: Forty-four patients had diabetes mellitus, 60 did not; half of each group had an epidural with continuous local anaesthetics. All patients received continuous insulin infusions during the initial 24 h period beginning with surgery. Blood glucose was measured four times daily (fasting or 2-3 h post-prandial) until end of the third postoperative day. RESULTS: For patients without diabetes, the epidural group had lower mean blood glucose and insulin requirements (P < 0.02) than controls during the initial 24 h period beginning with surgery. For patients with diabetes mellitus, thoracic epidural analgesia reduced mean blood glucose (P = 0.017) with unchanged insulin requirements. Epidural did not diminish the increase (vs. preoperative) in fasting blood glucose on the third postoperative day (32% vs. 22%, P < 0.001) for non-diabetics. Epidural analgesia was not able to attenuate hyperglycaemia during the first 3 postoperative days. CONCLUSIONS: Epidural analgesia improved glucose homeostasis minimally during the initial 24 postoperative hours but did not attenuate hyperglycaemia during the subsequent 3 postoperative days. PMID- 16045143 TI - Effects of mivacurium on the diaphragm evaluated by cervical magnetic stimulation of the phrenic nerves. AB - BACKGROUND AND OBJECTIVE: Non-depolarizing neuromuscular blocking agents have differential effects on the diaphragm and skeletal muscles. We employed a new method to study the effects of mivacurium on the diaphragm and compared the results obtained with this method with published data. METHODS: Anaesthesia was induced and maintained with propofol and alfentanil and the trachea was intubated after topical anaesthesia. Contractions of the diaphragm were induced by cervical magnetic stimulation of the phrenic nerves and quantified by measuring airway pressure responses. The neuromuscular effects on skeletal muscles were measured by acceleromyography of the adductor pollicis muscle. Mivacurium (0.15 mg kg(-1)) was injected and neuromuscular responses were recorded until the effects had waned. RESULTS: Eleven male and 10 female patients (ASA I-II; 57 +/- 16 yr; 78 +/ 13 kg; mean +/- standard deviation) participated. Median maximal reduction of twitch response was less (P < 0.05) for the diaphragm (89%) than for the adductor pollicis (100%). Time to 25% recovery was shorter for the diaphragm than for the adductor pollicis (8.8 +/- 2.2 min vs. 22.6 +/- 5.0 min, P < 0.05). The difference between the recovery index of the diaphragm (7.3 min (3.6-18.4)) and the adductor pollicis (8.2 min (4.4-20.9) (median (range)) just missed our chosen level of statistical significance (P = 0.06). The recovery time to train-of-four 0.8 was shorter for the diaphragm (median and 95% confidence interval 25.1 +/- 10.2 min) than for the adductor pollicis (median and 95% confidence interval 37.5 +/- 9.4 min, P < 0.05). CONCLUSIONS: The duration of the clinical effect of mivacurium on the diaphragm is markedly shorter than on the adductor pollicis muscles but there was only a small difference in the recovery index of the two muscles. These effects and the time courses determined with the new method closely resemble the results obtained with different methods in other studies. PMID- 16045144 TI - Partial liquid ventilation in acute salt water-induced lung injury. AB - BACKGROUND AND OBJECTIVES: Salt-water aspiration results in pulmonary oedema and hypoxia. We tested the hypothesis that partial liquid ventilation has beneficial effects on gas exchange and rate of survival in acute and extended salt water induced lung injury. METHODS: Anaesthetized, ventilated rats (tidal volume 6 mL kg(-1), PEEP 5 cmH2O) received a tracheal salt-water instillation (3%, 8 mL kg( 1) body weight) and were randomly assigned to three groups (n = 10 per group). While lungs of Group 1 were gas-ventilated, lungs of Group 2 received a single perfluorocarbon instillation (30 min after the injury, 5 mL kg(-1) perfluorocarbon) and lungs of Group 3 received an additional continuous perfluorocarbon application into the treachea (5 mL kg(-1) h(-1)) Arterial blood gases were measured with an intravascular blood gas sensor. RESULTS: Salt-water instillation resulted in a marked decrease in PaO2 values within 30 min (from 432 +/- 65 to 83 +/- 40 mmHg, FiO2 = 1.0, P < 0.01). Arterial oxygenation improved in all three groups irrespective of treatment. We observed no significant differences between groups in peak PaO2 and PaCO2 values. CONCLUSIONS: Our results suggest that partial liquid ventilation has no additional beneficial effects on gas exchange after life-threatening salt water-induced lung injury when compared to conventional gas ventilation with positive end-expiratory pressure. PMID- 16045145 TI - Terminal complement complex in septic shock with capillary leakage: marker of complement activation? AB - BACKGROUND AND OBJECTIVE: The aim of this study was to evaluate the value of terminal complement complex (C5b-9) plasma levels as a marker for complement activation in septic shock with concomitant capillary leak syndrome. METHODS: In a prospective animal study 10 fasted, anaesthetized, mechanically ventilated and multi-catheterized pigs (20.6 +/- 1.3 kg) were investigated over a period of 8 h. Sepsis was induced by faecal peritonitis (1 g kg(-1) body weight faeces, n = 5) and compared to controls (n = 5). The animals received 6% hydroxyethyl starch 200/0.5 to maintain a central venous pressure of 12 mmHg. To quantify capillary leak syndrome, albumin escape rate was measured using 99mTc-labelled human serum albumin. Plasma levels of terminal complement complex were measured in a double antibody immunoassay (neoepitope-specific MoAb aE 11 as catching antibody). Immunohistological studies of renal specimens were performed to detect terminal complement complex deposition. RESULTS: Albumen escape rate increased in septic animals (+ 52%) compared to controls (+ 3%, P < 0.05). Plasma levels of terminal complement complex decreased during the study period in both groups. In septic animals this finding was accompanied by a significant deposition of terminal complement complex in renal specimens (P < 0.05). CONCLUSION: We found an activation of the complement system proven by marked deposition of terminal complement complex in renal specimen, while its plasma levels decreased during the study period in septic and control animals. These results suggest that in septic shock with capillary leak syndrome plasma level of terminal complement complex may not be a reliable marker of complement activation. PMID- 16045146 TI - Use of recombinant factor VIIa for major haemorrhage. PMID- 16045147 TI - Nausea and vomiting after cataract surgery: does neostigmine have an emetic effect? PMID- 16045148 TI - Unexpected aortic dissection detected by transoesophageal echocardiography in the operating theatre at the beginning of cardiac surgery. PMID- 16045149 TI - Mediastinal haemorrhage mimicking tamponade during en-bloc oesophagectomy. PMID- 16045150 TI - Airway management in a patient with a vascular injury and rapidly expanding neck haematoma. PMID- 16045151 TI - Anaesthetic management of a patient with epidermolysis bullosa undergoing percutaneous nephrolithotomy. PMID- 16045152 TI - Percutaneous methanol intoxication: case report. PMID- 16045153 TI - An anaesthesia face mask with no lumen. PMID- 16045154 TI - Anaesthetic breathing circuit obstruction by a massive blood clot. PMID- 16045155 TI - Bridging the academy and the clinic. PMID- 16045156 TI - Is there a special psychoanalytic love? AB - Although the analyst's role mandates a degree of detachment, analysts have often said that they offer patients a special kind of love. They have tended to equate that love with understanding, thus neutralizing the paradox but also diluting the love. When something more resembling a loving affect is sought, the suggestions include the love a scholar feels for his subject, the love that accompanies immersion in great literature, and love that is self-generated by deliberate efforts to move toward the patient or to generate empathy in oneself. But a form of love may also arise from the analyst's unique and relatively pure acquaintance with a person's "appeal," by which is meant the patient's effort to elicit responses from the analyst. Although the awareness of transference most obviously tends to immunize him, it also gives him a poignant "insider's" feel of the patient's appeal, since he is the target of that appeal while being unencumbered by the myriad considerations that would color his perception were he to regard himself as a proper object. PMID- 16045157 TI - Analytic love and human vulnerability: a comment on Lawrence Friedman's "Is there a special psychoanalytic love?". PMID- 16045158 TI - Cordelia, Lear, and forgiveness. AB - Painful human interactions are often followed by urges to forgive, be forgiving, or seek forgiveness. The insight analysands develop into their transferences highlights their finding gratification in constantly reenacting painful interactions. Their new understanding can make forgiveness seem irrelevant; waiving the question of forgiveness might then seem the wiser course to follow. Also thrown into question is whether total forgiveness of self and others can ever be achieved. Shakespeare's The Tragedy of King Lear raises these questions. There we encounter, first, the painful interaction of Cordelia and Lear and, finally, Cordelia's response, "No cause, no cause," to a dying Lear's begging her forgiveness for having initially treated her cruelly. Cordelia's response seems to be waiving the question of guilt and forgiveness, but could it be wholehearted? In a search for answers, a reading of Cordelia's and Lear's lines is interwoven with interpretations of unconscious conflict that might be considered were one to encounter clinically a "Cordelia" abused by an aging and failing father at a turning point in her womanly development. Unconsciously, it is concluded, unforgivingness persists alongside the loving, insightful waiving of forgiveness made possible by higher-level ego functioning. Methodological reflections on reading and interpretation are included. PMID- 16045159 TI - Pleasures of mind. PMID- 16045160 TI - The impossibility of forgiveness: shame fantasies as instigators of vengefulness in Euripides' Medea. AB - Unforgivability in Euripides' Medea is explored in the context of intrapsychic forces favoring disruption and narcissistic withdrawal and precluding the influence of forces favoring repair of bonds, not necessarily to the betrayer, but to the social and moral order. The forces underlying disruption and withdrawal operate to such an extent that forgiveness and cooperation with the social order become impossible. Euripides' literary insights are explored with the purpose of deepening and extending the psychoanalytic understanding of shame, shame fantasies, projective identification, and vengefulness as they bear on the problem of forgiveness. Three types of shame fantasy are pertinent to the transformation of Medea's mental state from one of anguished and disjointed shame to diabolical vengefulness: anticipatory paranoid shame, the projective identification of shame, and withdrawal as a defense against shame. PMID- 16045161 TI - Shame conflicts and tragedy in The Scarlet Letter. AB - Nathaniel Hawthorne's The Scarlet Letter has much to teach psychoanalysts and psychotherapists. Perhaps no other American novel lends itself so well to an exploration in depth of the dynamics, conflicts, and defenses characteristic of shame. While most commentators on The Scarlet Letter have assumed Hester Prynne's pain to be shame-based, and the Reverend Dimmesdale's to be guilt-based, a rather different interpretation is proposed namely, that both are afflicted with shame, but that Dimmesdale's is more unbearable than Hester's because more conflictual, less representable, and less easily used protectively. Dimmesdale's shame is at once deeper and more toxic. What "deeper" and "more toxic" mean in the context of shame conflicts (including conflicts to which feelings of shame give rise) is explored. PMID- 16045162 TI - The capacity to forgive: intrapsychic and developmental perspectives. AB - The topic of forgiveness, despite its importance for the continuity of relationships and for the mental health of the aggrieved party, is relatively neglected in the psychoanalytic literature, perhaps because it is often seen as the province of religion and carries the connotation of reaction formation and inauthenticity. However, genuine forgiveness involves significant intrapsychic work, conscious and unconscious working through of one's anger, and putting the offense into the context of an integrated view of the whole person of the offender. Early developmental structures are the ground on which the relative ability to let go of a grievance depends. While later motives and defenses (e.g., fear of retraumatization, avoidance of shame) may also play a role, these early structures are primary. They are described here in terms of attainment of the depressive position and the development of a sense of secure attachment, the capacity to mentalize, and the ability to mourn. PMID- 16045164 TI - The social face of shame and humiliation. Panel report. PMID- 16045163 TI - Revenge. Panel report. PMID- 16045165 TI - The legacy of Enrico Jones. PMID- 16045166 TI - On analytic process. AB - An innovative methodology is presented for identifying and assessing change process in psychoanalytic treatments. Using the Psychotherapy Process Q-set (PQS), a panel of experienced psychoanalysts developed a prototype of an ideal psychoanalytic hour. This prototype was then applied to verbatim transcripts of three archived treatment samples: psychoanalyses, long-term analytic therapies, and brief psychodynamic therapies. The degree to which these treatments fostered an analytic process as represented by the prototype was measured quantitatively. Analytic process was significantly more present in psychoanalyses than in the long-term analytic therapies, which, in turn fostered significantly more analytic process than did brief psychodynamic therapies. The study demonstrates that, given descriptive language that does not represent a particular theoretical perspective, analysts can agree on a definition of analytic process, and that analytic process can be operationalized and quantitatively assessed. A second study demonstrates that despite consensus on its definition, there is not just one proper analytic process; rather, there are change processes unique to each dyad. Two quantitative case studies illustrate how each analytic pair has a unique interaction pattern linked to treatment progress. These dyad-unique "interaction structures" are recurrent, mutually influencing patterns of interaction, the experience, recognition, and comprehension of which appear to be a fundamental component of therapeutic action. A bipersonal model is described that attempts to bridge theories of therapeutic action that focus on insight and self-understanding and those that emphasize the patient's experience of the therapist. PMID- 16045167 TI - In praise of simplicity: commentary on Ablon and Jones. PMID- 16045168 TI - Empirical evidence supporting the conceptual relatedness of object representations and internal working models. AB - The proliferation of new developmental and clinical theories broadly termed "psychoanalytic" demands a methodology for making systematic comparisons to establish the commonalities and distinctions among them. This article presents an empirical method for the comparative evaluation of such theories. The Mother-To Child Object Representation / Internal Working Model Q-sort is a 100-item instrument constructed to assess the quality of the prototypical mother's object representation and internal working model of her child at age five. Object relations judges were asked to sort these items for a complex, differentiated, integrated object representation of a five-year-old, attachment judges for a secure, coherent, freely valuing internal working model of a five-year-old. Judges' criterion Q-sorts in each group yielded a composite criterion Q-sort of the quality of the prototypical mother's mental representation of her child according to each group's theoretical perspective. The Spearman-Brown correlation between the two composite criterion Q-sorts was r = .90, p < .001 , suggesting that when confined to a 100-item common vocabulary, judges representing each theoretical construct agreed on the conceptual relatedness of object representations and internal working models. The theoretical constructs of object representations and internal working models share common assumptions in need of further exploration. PMID- 16045169 TI - Confoundng factors in research populations. PMID- 16045170 TI - [New insecticides--focusing phenylpyrazol derivatives]. PMID- 16045171 TI - [Psychiatric aspects of acute intoxicated patients]. PMID- 16045172 TI - [Lethal suicide attempts and parasuicide in drug overdose patients]. PMID- 16045173 TI - [Self-poisoning repeater--psychiatric considerations]. PMID- 16045174 TI - [Triage guideline for suicidal patients with acute poisoning and drug overdoses]. PMID- 16045175 TI - Case with bromine exposure leading to respiratory insufficiency. AB - A 21-year-old male had a chemical burn on the right forearm when he inadvertently spilled bromine during an experiment. Since he inhaled vaporized bromine and had dyspnea and pharyngalgia, he arrived at our hospital in an ambulance as an emergency patient. On arrival, he kept a clear consciousness with a pulse rate of 98, body temperature of 36.8 degrees C, blood pressure of 132/80 mmHg, respiratory rate of 25, and oxygen saturation of 100%. (10 L/min of oxygen were administered.) He had marked dry coughs. His clothes had a foreign odor with mucosal irritation. Arterial blood gas analysis and blood biochemistry were normal. Based on these findings, he was diagnosed with chemical airway damage and bulbar conjunctiva from the exposure to bromine and a chemical burn on the right forearm. His respiratory condition became worse after admission, resulting in pulmonary edema. He was endotracheally intubated and controlled with an artificial ventilator on Day 3 after his injury. He was continuously treated with steroids and sivelestat sodium hydrate, which gradually improved his respiration. He was released from the artificial ventilator and extubated on Day 7. Although dyspnea associated with body movement and hoarseness persisted after extubation, the symptoms decreased and he was discharged on Day 41. This rare case is worth attention because serious respiratory insufficiency requiring artificial ventilation due to pulmonary edema from bromine exposure has not been reported in Japan. PMID- 16045176 TI - [Case of accidental ingestion of caster beans: acute intoxication by ricin]. PMID- 16045177 TI - [Guidelines for the treatment of acute chemical poisoning--9--symptomatic therapy. Body temperature management]. PMID- 16045178 TI - [Practical analysis of toxic substances useful for clinical toxicology--14- arsenics]. PMID- 16045179 TI - [2004 report on North American Congress of Clinical Toxicology (NACCT)]. PMID- 16045180 TI - [2004 Annual Report by JPIC]. PMID- 16045181 TI - [Hospital rankings by number of surgeries ]. PMID- 16045182 TI - [Treatment strategies for pediatric solid tumors]. PMID- 16045183 TI - [Treatment strategy for malignant solid tumors in childhood]. AB - The prognosis for children with malignant solid tumors has improved dramatically in Japan. During the last two decades, various groups have conducted sequential studies of the treatment of children with neuroblastoma, Wilms' tumor, and hepatoblastoma. Most institutes participated in nonrandomized trials designed to evaluate the safety and efficacy of combination chemotherapy, surgery, and radiotherapy in each group study and treated children with these tumors The results are reviewed and areas for future investigation are identified. PMID- 16045184 TI - [Past and future role of surgery in neuroblastoma]. AB - The significance of surgery in the treatment of neuroblastoma remains unresolved. Moderate surgical resection has been employed in both infantile and advanced neuroblastoma since 1985 at the University of Tsukuba Hospital. Resection of the original tumor accompanied by lymph node sampling was performed in 39 cases of infantile neuroblastoma detected by mass screening. The patients are alive without tumor and with no treatment sequelae. In 20 patients with advanced neuroblastoma aged 1 year or older, the same surgery was performed with intraoperative radiation after more than 5 cycles of induction chemotherapy. Four patients experienced local recurrence with recurrence of remote metastases which were the cause of patient death. The overall survival rate of patients with stage 4 neuroblastoma was 70% at 5 years and 62% at 10 years. A clinical trial in which local therapy will be performed after myeloablative chemotherapy will be carried out from 2005 to evaluate separately the efficacy of chemotherapy and local therapy avoiding their direct interference. This will elucidate the value of local therapy and also moderate surgical resection in the treatment of advanced neuroblastoma. PMID- 16045185 TI - [Treatment strategy for children with Wilms tumor]. AB - Wilms tumor has an excellent prognosis according to the findings of the National Wilms Tumor Study (NWTS). The NWTS has been aiming to reduce the intensity of treatment for low-risk groups. Pathologic assessment or statistical analysis has identified the appropriate staging system suited to this treatment regimen. The Japanese Wilms Tumor Study (JWiTS) has been following the NWTS-5 regimen to increase survival rates and tumor-free survival rates from 1996. This study aims to establish a central review system of pathology and to study the presence of WT1 mutation in tumor samples. In addition, JWiTS focuses on developing an online database that can be shared over the Internet. However, the prognosis in patients with malignant rhabdoid tumor of the kidney (MRTK) is still poor. JWiTS has started to consider a new regimen for MRTK. PMID- 16045186 TI - [Strategy for the treatment of hepatoblastoma in children]. AB - Hepatoblastoma is the most common malignant live tumor in childhood. Multicenter trials have made it clear that pre- and/or postoperative chemotherapy are necessary to improve the outcome of the treatment of hepatoblastoma. The results of the Japanese Pediatric Liver Tumor (JPLT)-1 study since 1991 have been compatible with those of other group study trials in Europe and the USA. However, patients with unresectable primary tumors or with lung metastasis still have a poor prognosis. The JPLT has developed a new protocol that includes four courses of treatment based on the PRETEXT staging system. These treatments include intensive chemotherapy with stem cell transplantation for patients with unresectable primary tumors or with pulmonary metastases. PMID- 16045187 TI - [Childhood rhabdomyosarcoma]. AB - Overall five-year survival of Childhood rhabdomyosarcoma (RMS) is reported to be 70% in the Intergroup Rhabdomyosarcoma Study Group (IRSG), however, the figure in Japan is almost 15% lower than that of IRSG. Treatment regimen of RMS essentially depends on the histology of the tumor, site, preoperative staging and postoperative grouping that leads to the risk classification. VAC is a standard chemotherapeutic regimen for low and intermediate risk group. Clinical trial with CPT-11 or high dose chemotherapy are underway for high risk RMS. PMID- 16045188 TI - [Treatment for Ewing's sarcoma family of tumors in children]. AB - The outcome of patients with Ewing Sarcoma Family of Tumors (ESFT) in our country is poor in comparison with American and European. We performed a review of the relevant data regarding the current status in the treatment of ESFT. The results of the major American and European cooperative groups were discussed, including the treatment strategies used and the prognostic factors identified for both localized and metastatic ESFT. We also discussed a new trial for the establishment of the standard treatment of ESFT in our country. PMID- 16045189 TI - [Surgical stress]. PMID- 16045190 TI - [What we can learn from a case of medical malpractice]. PMID- 16045191 TI - [Management of unresectable gastric cancer with perforation]. PMID- 16045192 TI - [Management of unresectable case of perforated rectal cancer]. PMID- 16045193 TI - [Experimental behavioral tests using monkey and rat offspring born from mothers exposed perinatally to EDCs]. AB - Purpose of this study is to conduct risk assessment of EDCs for the development of CNS in humans by extrapolation from the results of behavioral tests in rats and monkeys. Our hypotheses on the mechanism which gives an adverse effect of EDCs to the developing neural systems are as follows. Thyroid hormone (TH) disrupting chemicals induce deterioration of neural development and estrogen (E2) agonistic chemicals may disturb apoptosis of fetal neural cells resulting in injury of normal neural circuit. The strategy of this study is a bottom up system; for example, basic information was obtained by an experiment using rats and then an experiment using monkey was designed to adapt the results from rats. The monkey experiment data will be assessed in comparison with human behavior. The tactics of this study are, however, a top down system. It is neural behaviors which are an end point evaluation that are primarily performed. They are mother infant interactions, social behaviors, open field test, memory and learning tests, etc. As for analysis of the mechanism of EDCs' adverse effect, we tried two methods: one is an in vivo drug biased test which interferes with the monoamine oxidase (MAO) system and the other is an in vitro primary neural cell culture. EDCs including BPA, NP, propylthiouracil (PTU) and PCB-OH are injected orally to pregnant rats from gestation day 3 (GD3) to post natal day 21 (PND21) at weaning and their offspring were tested. On the other hand TCDD, BPA and PCB effect are assessed in rhesus monkey or cynomolgus monkey offspring. The study is still continuing and we will present the results obtained to date. PMID- 16045194 TI - [Exposure to bisphenol-A affects the rewarding system in mice]. AB - Bisphenol-A has been extensively evaluated for toxicity in a variety of tests as the most common environmental endocrine disruptor. In the present study, we found that prenatal and neonatal exposure to bisphenol-A affects the development of the central dopaminergic system in the mouse limbic area. Additionally, this treatment with bisphenol-A produced a down-regulation of dopamine D3 receptor and an up-regulation of dopamine D1 receptor function to activate G-protein in the mouse limbic forebrain, which is thought to play a critical role for rewarding effects by drugs of abuse. We next investigated the relationship between the neurobehavioral toxicity and its exposure period. The exposure to bisphenol-A during either organogenesis or lactation, but not implantation and parturition, significantly enhanced the morphine-induced hyperlocomotion and rewarding effect. Furthermore, the exposure to bisphenol-A during either organogenesis or lactation also produced an up-regulation of dopamine D1 receptor function to activate G protein in the mouse limbic forebrain. These results indicate that either organogenesis or lactation is more sensitive to the bisphenol-A-induced neuronal toxicity than any other periods. In conclusion, we found here that prenatal and neonatal exposure to bisphenol-A can potentiate the central dopaminergic systems, resulting in the supersensitivity of the drugs of abuse-induced rewarding effects and hyperlocomotion in the mouse. Furthermore, the organogenesis and lactation are the most important period to cause the alteration of dopaminergic system by bisphenol-A exposure in the mouse. PMID- 16045195 TI - [Clinical pharmacogenetics in the treatment of schizophrenia]. AB - Numerous investigations on metabolic enzymes, cytochrome P450 (CYP), have been conducted since 1990. In the psychiatric field, the focus has been on CYP2D6, which is a major enzyme involved in metabolism of antidepressants and antipsychotics. Poor metabolizers (deficit metabolizers) for CYP2D6 represent 7% among Caucasians, while they accouut for less than 1% of Asians. The frequency of a mutated allele for CYP2D6*10, which leads to the decrease in CYP2D6 activity, is 40% in Asians. It has been reported that steady-state plasma concentration of haloperidol in subjects with mutated alleles for CYP2D6 is significantly higher than that in subjects without mutated alleles. At the same time, steady-state plasma concentration of risperidone is very different between CYP2D6 genotypes. In addition, several studies suggest that better efficacy or higher scores of side effects are observed in the subjects with mutated alleles for CYP2D6. Recently receptor polymorphism has become a concern and association between clinical response and polymorphism of dopamine and serotonin has been reported. In the dopamine D2, subjects with -141C Ins allele in -141C Ins/Del polymorphism and subjects with A1 allele in Taq1A have a better response to dopamine antagonists. Association between Ser allele and typical antipsychotics and between Gly allele and atypical antipsychotics has been investigated. There are still no data indicating significant association between dopamine D1 and clinical response to antipsychotics. Clinical pharmacogenetical studies from both a pharmacokinetical and a pharmacogynamical point of view are required in order to introduce and practice individualized medicine in psychiatric field easily. PMID- 16045196 TI - [In vivo evaluation of expressed protein function by PET]. AB - Positron Emission Tomography (PET) allows in vivo visualization of the expression and function of protein using a radioligand. Quantitative analysis of serotonin transporter, receptors, and the function of P-Glycoprotein has been performed in living human brains. Furthermore, the relationship between the phenotype of those proteins and their genetic polymorphism has also been investigated. Regarding the effect of antipsychotics on dopamine D2 receptor, occupancy and its time-course have been measured in a living body using PET. This approach can provide in vivo pharmacological evidences of antipsychotics and establish a rational therapeutic strategy. PET is a powerful tool not only in the field of brain research but also drug discovery and individual medicine. PMID- 16045197 TI - [Pharmacological profiles of benzodiazepinergic hypnotics and correlations with receptor subtypes]. AB - We examined the behavioral pharmacological properties of six benzodiazepine (omega) receptor ligands including brotizoram, nitrazepam, quazepam, rilmazafone, zolpidem and zopiclone and the binding of these drugs with omega receptor subtypes. Behavioral tests were performed at the time of the maximal effects induced by each drug following its oral administration to mice. All of these drugs dose-dependently induced impairment of motor coordination as rotarod performance and potentiation of thiopental-induced anesthesia as hypnotic effect. The hypnotic effects of rilmazafone, whose major metabolites were bound to both omega1 and omega2 receptors with high affinity, and omega1 selective quazepam were about 20 times more effective than the induction of motor impairments when compared with ED50 values. However, there was no difference between the ED50 values of omega1 selective zolpidem alone in these two tests. An antianxiety efficacy of zolpidem was relatively weak unlike that of other drugs in the elevated plus-maze. It has been reported that omega2, but not omega1, receptors are associated with motor impairment and anxiolytic effect. The weak anxiolytic effect of zolpidem supports the previous hypothesis. However, the strong motor incoordination of zolpidem suggests that not only omega2 but also omega1 receptors are related to motor impairment unlike the previous hypothesis. PMID- 16045198 TI - [Behavioral characterization of mouse strains in learning and memory tests]. AB - Here we review the behavioral characterization of several strains of mice that have generally been used in learning and memory tests. We made behavioral comparisons among inbred (C57BL/6, DBA/1 and 129Sv) and closed colony (ddY and ICR (CD-1)) mice. In the water-maze test, the spatial learning ability of C57BL/6 and ICR was higher than that of the other strains of mice. In the Y-maze, water finding and step-through type passive avoidance tests, there were some differences among those strains but they were not significant. We have to pay attention to the mouse strain to analyze the behavioral changes when we use different strains of mice. PMID- 16045199 TI - [Electrogenic Na+/Ca2+-exchange regulation by alpha2-receptor in peripheral sympathetic nerve]. AB - Electrical depolarisation-(2 Hz, 1 ms)-induced [3H]noradrenaline ([3H]NA) release was measured from the isolated main pulmonary artery of the rabbit in the presence of uptake blockers (cocaine, 3 x 10(-5)M; corticosterone, 5 x 10(-5)M) and after blocking the MAO-enzyme by pargyline (1,2 x 10(-4)M). Substitution of most of the external Na+ by Li+ (113 mM; [Na+]0: 25 mM) slightly potentiated the stimulation-induced release of [3H]NA in a tetrodotoxin (TTX, 10(-7)M) sensitive manner. The reverse Na+/Ca2+-exchange inhibitor KB-R7943 (3 x 10(-5)M) failed to inhibit the stimulation-evoked release of [3H]NA, but increased the resting outflow of neurotransmitter. The 'N-type' voltage-sensitive Ca2+-channel (VSCC) blocker omega-conotoxin (omega-CgTx) GVIA (10(-8)M) significantly and irreversibly inhibited the release of [3H]NA on stimulation (approximately 60 70%). The 'residual release' of NA was abolished either by TTX or by reducing external Ca2+ from 2,5 to 0,25 mM. The 'residual release' of NA was also blocked by the non-selective VSCC-blocker neomycin (3 x 10(-3)M). Direct correlation was obtained between the extent of VSCC-inhibition and the transmitter release enhancing-effect of presynaptic alpha2-receptor blocker yohimbine (3 x 10(-7)M). When the release of [3H]NA was blocked by omega-CgTx GVIA plus neomycin, yohimbine was ineffective. Inhibition of the Na+-pump by removal of K+ from the external medium increased both the resting and the stimulation-evoked release of [3H]NA in the absence of functioning VSCCs (i.e. in the presence of neomycin and after omega-CgTx treatment). Under these conditions the stimulation-evoked release of NA was abolished either by TTX or by external Ca2+-removal (+1 mM EGTA). Similarly, external Li+ (113 mM) or the reverse Na+/Ca2+ exchange blocker KB-R7943 (3 x 10(-5)M) significantly inhibited the nerve-evoked release of NA in 'K+-free' solution. KB-R7943 decreased the resting outflow of NA as well. Under conditions, in which the Na+-pump was inhibited in the absence of functioning VSCCs, yohimbine (3 x 10(-7)M) further enhanced the release of neurotransmitter, while l-noradrenaline (l-NA, 10(-6)M), an agonist of presynaptic alpha2 receptors, inhibited it. The yohimbine-induced enhancement of NA-release was abolished by Li+-substitution and significantly inhibited by KB-R7943 application. It is concluded that after blockade of VSCCs, brief depolarising pulses may reverse Na+/Ca2+-exchange and release neurotransmitter in Na+-loaded sympathetic nerves. Further, similar to that of VSCCs, the reverse Na+/Ca2+ exchange may also be inhibited by presynaptic alpha2-receptor activation. PMID- 16045200 TI - [Monitoring the temperature distribution during dielectric drying]. AB - Microwave assisted vacuum drying offers never questioned advantageous and it is getting more and more popular lately. In spite of its uniqueness there is a rightful resistance and mistrust because the temperature distribution of the workload is far from being homogeneous as a result of the existing inhomogeneous electromagnetic field. The aim of this study is to survey the heat thus field distribution in the workload to prevent local overheating that endangers the quality of the pharmaceutical product. Thermography as a non perturbing thermometrical method was used to map the heat distribution in the workload. To get a 3D information Teflon layers were used to form cross-sectional and photogenic surfaces about the dusty blend and a purpose-developed software to evaluate qualitatively the IR pictures. PMID- 16045201 TI - [Particle size determination using the method of dynamic light scattering]. AB - Realising the lacks of the classic methods of particle size determination a new method was needed to be developed which became suitable for measurements in the nano range as well. The principle of this method was discovered more than 20 years ago but manufacturers are still able to insert novelties by developing and refining the technique of - dynamic light scattering. Importance of multiparticular systems increases in pharmaceutical technology. It is necessary to apply this new method to investigate them. PMID- 16045202 TI - [Investigation of solubility properties of nifluminic acid containing cyclodextrins and polyvidone]. AB - One of the most important task of the pharmaceutical technology is to reach a possible high gastrointestinal absorbtion of the active ingredient. Therefore, it is necessary to have a good solubility in the gastric/intestinal medium. In this way the applied drug quantity and unwanted side effects can be reduced with solubility increasing. The water-insoluble drug's solubility and bioavailability can be increased by the alteration of their physicochemical properties. Nifluminic acid is a frequently used anti-inflammatory drug with low aqueous solubility. Inclusion complexation with cyclodextrins is an efficacious method to improve the solubility, stability and bioavailability. Ternary systems were prepared and investigated to use nifluminic acid, hydoxypropyl-beta-cyclodextrin and polyvidone. The aims of the study were to investigate the solubility rate, the in vitro diffusion ability, to determine the n-octanol/water partition coefficient, and to study the thermoanalitycal properties of products. The results suggested that complexation with cyclodextrin and the use of polyvidone is better method than to prepare binary systems. Inclusion complexation was presumed in the event of the ternary composition to improve the bioavailability of nifluminic acid. PMID- 16045203 TI - [Pharmaceutical chemistry of neuroleptic drugs]. AB - The paper represents the 3rd part of a series about agents acting on the central nervous system. This time the neuroleptics are surveyed. As previously, the material is divided into chapters of history, preparation; structure-properties activity; therapeutical use; analysis. PMID- 16045204 TI - Drifts exhibited by cryogenically cooled InSb infrared filtered detectors and their importance to the ATSR-2 and Landsat-5 Earth observation missions. AB - The spectral responsivity of commercially available InSb detectors with low-pass cold filters attached to their cold shields for optimum operation in the 1.6-2.6 microm wavelength range is observed to drift slowly with time. These drifts are shown to arise because of a thin film of water-ice deposited on the cold low-pass filters mounted on the cold shields of the detectors. The temporal characteristics of these drifts are shown to strongly depend on wavelength. A model is proposed for the behavior of the water present in the Dewar vacuum, which can explain and predict the temporal characteristics of the observed drifts for all wavelengths. These observations are particularly relevant to space instruments that use cryogenically cooled IR filter radiometers for Earth observation. The temporal profile of drifts observed in missions such as Landsat 5 is identical to that observed in cryogenically cooled filtered InSb detectors during laboratory measurements. This study confirms that the deposition of a thin film of a material such as ice on the cold bandpass filters and windows is therefore the most likely source of the oscillatory drifts observed in the response of some of the channels of the ATSR-2, Landsat-4, and Landsat-5 Earth observation missions. PMID- 16045205 TI - Sensitivity metric approach for retrieval of aerosol properties from multiangular and multispectral polarized radiances. AB - Linearly polarized radiation is sensitive to the microphysical properties of aerosols, namely, to the particle-size distribution and refractive index. The discriminating power of polarized radiation increases strongly with the increasing range of scattering angles and the addition of multiple wavelengths. The polarization and directionality of the Earth's reflectances (POLDER) missions demonstrate that some aerosol properties can be successfully derived from spaceborne polarimetric, multiangular measurements at two visible wavelengths. We extend the concept to analyze the retrieval capabilities of a spaceborne instrument with six polarimetric channels at 412, 445, 555, 865, 1250, and 2250 nm, measuring approximately 100 scattering angles covering a range between 50 and 150 deg. Our focus is development of an analysis methodology that can help quantify the benefits of such multiangular and multispectral polarimetric measurements. To that goal we employ a sensitivity metric approach in a framework of the principal-component analysis. The radiances and noise used to construct the sensitivity metric are calculated with the realistic solar flux for representative orbital viewing geometries, accounting for surface reflection from the ground, and statistical and calibration errors of a notional instrument. Spherical aerosol particles covering a range of representative microphysical properties (effective radius, effective variance, real and imaginary parts of the refractive index, single-scattering albedo) are considered in the calculations. We find that there is a limiting threshold for the effective size (approximately 0.7 microm), below which the weak scattering intensity results in a decreased signal-to-noise ratio and minimal polarization sensitivity, precluding reliable aerosol retrievals. For such small particles, close to the Rayleigh scattering limit, the total intensity provides a much stronger aerosol signature than the linear polarization, inspiring retrieval when the combined signals of intensities and the polarization fraction are used. We also find a strong correlation between aerosol parameters, in particular between the effective size and the variance, which forces one to simultaneously retrieve at least these two parameters. PMID- 16045206 TI - Noncontact electrical test of a ball grid array substrate that uses the electro optic probing technique. AB - A new technique for testing a ball grid array (BGA) package substrate that uses the electro-optic (EO) probing technique is investigated. This technique can detect open circuits in the BGA substrate with a high spatial resolution. An experimental setup that uses an EO probe tip made of LiNbO3 crystal is reported along with the measurement results from a real BGA substrate. PMID- 16045207 TI - Assessment of soot particle vaporization effects during laser-induced incandescence with time-resolved light scattering. AB - Although laser-induced incandescence (LII) has been successfully used for soot volume fraction and particle size measurements, uncertainties remain regarding issues of soot vaporization leading to mass loss and morphological changes occurring in soot due to intense heating. Prompt LII detection schemes are often based on the assumption that the associated time scale is shorter than the time scale of soot vaporization or sublimation. The validity of such assumptions is the focus of the current study. Time-resolved light-scattering measurements were made in combination with LII measurements to quantify soot particle vaporization effects resulting from the LII laser pulse. The light-scattering measurements revealed a sharp decrease in total soot particle mass during the time course of the 25 ns full-width LII laser pulse for fluences in the range of 0.5 J/cm2. Light-scattering theory was used to invert the scattering data, revealing approximately 80%-90% reductions in the soot particle volume for LII fluences of 0.47 and 0.61 J/cm2. In addition, the time-resolved scattering measurements show that the time scale of soot vaporization is completely confined to the LII laser pulse itself. Light scattering revealed no soot vaporization only for fluences of approximately 0.1 J/cm2, which is consistent with recent work on low-fluence LII. Possible mechanisms for soot vaporization are discussed, notably for near threshold fluences. PMID- 16045208 TI - Effects of index-mismatch-induced spherical aberration in pump--probe microscopic image formation. AB - Pump--probe fluorescence microscopy has been demonstrated to be a powerful tool for obtaining three-dimensional, time-resolved information in bioimaging applications. However, the use of this technique can be complicated by the fact that the different wavelengths used to achieve pump--probe microscopy can result in wavelength-dependent spherical aberration, thus limiting the usefulness of the technique. We address this issue by investigating the effects of refractive-index mismatch-induced spherical aberration on pump--probe image formation. We model the effects by considering pump--probe imaging performed with an objective with a numerical aperture of 0.75 focusing through an oil-water interface. Our results show that spherical aberration has the greatest effect in degrading an axial point-spread function. In addition to signal loss, the redistribution of signal strength along the axial direction results in broadening of the FWHM of the plane response function. The inclusion of confocal detection tends to improve image resolution but at a significant loss of signal strength. PMID- 16045209 TI - Wave-front retrieval from Hartmann test data. AB - In the classical Hartmann test the wave front is obtained by integration of the transverse aberrations, joining the sampled points by small straight segments, in the so-called Newton integration. This integration is performed along straight lines joining the holes on the Hartmann screen. We propose a modification of this procedure, considering the cells of four holes of the Hartmann screen to fit a small second-power wave front recovering each square. This procedure has some important advantages, as described here. PMID- 16045210 TI - Use of null optics for monitoring the optical alignment of a beam director. AB - The use of null optics is proposed as a new concept for monitoring the optical alignment of a beam director. Null optics consist of a primary mirror and an annular mirror just outside the aperture of the secondary mirror. The characteristics of the proposed null optics are investigated with the designed sample of a two-mirror system with an effective aperture of 275 mm. The results show that null optics yield four times the amplification with respect to the alignment errors of the secondary mirror; however, they have low residual aberration sensitivity to misalignment. Therefore null optics can be used successfully as the alignment monitoring apparatus of a beam director. PMID- 16045211 TI - Collimation testing with optically active materials. AB - A novel method to test the collimation of laser beams with optically active mediums and a pair of crossed polarizers is presented. Optically active materials rotate the plane of polarization of incident plane-polarized light. A decollimated laser beam passing through such a material will experience a greater effective thickness than a collimated laser beam, resulting in greater outputs. In this method the output intensity variation is related to the amount of decollimation of the incident beam, and the method does not require any referencing or fringe analysis and is easy to implement. PMID- 16045212 TI - Holographic polarization-selective module based on a small dove prism coupler for magneto-optical pickup heads. AB - A new structure for polarization-selective elements, consisting of two holographic gratings and a Dove prism coupler, is proposed. The absence of a multistage waveguide and the benefits of compact size and lightweight volume are the outstanding features of the new structure. Based on the coupled-wave theory, the analysis and design of the structure are discussed in detail to calculate the required index modulation. Several parameters, such as the recording intensity, the exposure time, and the recording angles for the fabrication of the proposed element, are determined. Under these conditions, the element is fabricated in Dupont photopolymer HRF-150-38 material and with an operating wavelength of 532 nm. A simplified pickup head is constructed to evaluate the performance of the fabricated element. PMID- 16045213 TI - Estimation of chest-wall-induced diffused wave distortion with the assistance of ultrasound. AB - The chest-wall layer underneath breast tissue consists of muscles and bones, which induce distortion in near-infrared diffused waves measured at distant source--detector pairs when reflection geometry is used. A priori information on chest-wall depth obtained from coregistered real-time ultrasound can be used to assist in the removal of distant measurements. We applied Monte Carlo simulation to a simple two-layer model consisting of breast tissue and a chest wall to investigate chest-wall-induced distortion. The Monte Carlo method indicates that, when more than 50% of the received photons travel through the breast tissue layer before being detected, the detected signal may be useful for image reconstruction. The results of phantom experiments obtained from the two-layer model further validate the distortion problem and demonstrate imaging improvement after distant measurements have been filtered out. Clinical examples have shown similar imaging improvements on reconstructed absorption maps. Clinical data obtained from 20 patients with the chest-wall depths of less than 2 cm from the skin surface suggest that the cutoff distances of distorted measurements are largely related to the chest-wall depth and are relatively independent of the optical properties of tissue. PMID- 16045214 TI - Transillumination optical tomography of tissue-engineered blood vessels: a Monte Carlo simulation. AB - A Monte Carlo technique has been developed to simulate the transillumination laser computed tomography of tissue-engineered blood vessels. The blood vessel was modeled as a single cylinder layer mounted on a tubular mandrel. Sequences of images were acquired while rotating the mandrel. The tomographic image was reconstructed by applying a standard Radon transform. Angular discrimination was applied to simulate a spatial filter, which was used to reject multiply scattered photons. The simulation results indicated that the scattering effect can be overcome with angular discrimination because of the thin tissue thickness. However, any refractive-index mismatch among the tissue, the surrounding media, and the mandrel could produce significant distortions in the reconstructed image. PMID- 16045215 TI - Dispersion compensation in high-speed optical coherence tomography by acousto optic modulation. AB - We report studies of the analyses of and compensation for group dispersion to improve the axial resolution of high-speed optical coherence tomography (OCT) by acousto-optic modulation (AOM). Theoretical modeling and experiments reveal that the high-order group dispersion induced by acousto-optic crystals broadens the measured coherence length (Lc) and thus degrades the axial resolution of OCT imaging. Based on our experimental studies, we can compensate for the dispersion to less than 50% broadening of the source Lc by adjusting the grating-lens-based optical delay in the reference arm and can further eliminate it by inserting like acousto-optic crystals in the sample arm of the OCT system. The results demonstrate that this AOM-mediated OCT system permits high-performance OCT imaging at A-scan rates of as much as 4 kHz by use of a resonant scanner. Because of its ultrastable direct frequency modulation, this AOM-mediated OCT system can potentially improve the performance of high-speed Doppler OCT techniques. PMID- 16045216 TI - Real-time absorption and scattering characterization of slab-shaped turbid samples obtained by a combination of angular and spatially resolved measurements. AB - We present a fast and accurate method for real-time determination of the absorption coefficient, the scattering coefficient, and the anisotropy factor of thin turbid samples by using simple continuous-wave noncoherent light sources. The three optical properties are extracted from recordings of angularly resolved transmittance in addition to spatially resolved diffuse reflectance and transmittance. The applied multivariate calibration and prediction techniques are based on multiple polynomial regression in combination with a Newton--Raphson algorithm. The numerical test results based on Monte Carlo simulations showed mean prediction errors of approximately 0.5% for all three optical properties within ranges typical for biological media. Preliminary experimental results are also presented yielding errors of approximately 5%. Thus the presented methods show a substantial potential for simultaneous absorption and scattering characterization of turbid media. PMID- 16045217 TI - Reflectance spectroscopy for diagnosis of epithelial precancer: model-based analysis of fiber-optic probe designs to resolve spectral information from epithelium and stroma. AB - Reflectance spectroscopy is a promising technology for detection of epithelial precancer. Fiber-optic probes that selectively collect scattered light from both the epithelium and the underlying stroma are likely to improve diagnostic performance of in vivo reflectance spectroscopy by revealing diagnostic features unique to each layer. We present Monte Carlo models with which to evaluate fiber optic probe geometries with respect to sampling depth and depth resolution. We propose a probe design that utilizes half-ball lens coupled source and detector fibers to isolate epithelial scattering from stromal scattering and hence to resolve spectral information from the two layers. The probe is extremely compact and can provide easy access to different organ sites. PMID- 16045218 TI - Fiber-optic localization by geometric space coding with a two-dimensional gray code. AB - With the objective of monitoring motion within a room, we segment the two dimensional (2D) floor space into discrete cells and encode each cell with a binary code word generated by a fiber. We design a set of k-neighbor-local codes to localize an extended object and, particularly when k = 2, employ a 2D gray code to localize a human by tracking his or her footsteps. Methods for implementing the codes in a fiber web are discussed, and we demonstrate the experimental result with the fiber mat. The observed system performance confirms the theoretical analysis. The space coding technique is a promising low-cost candidate not only for human tracking but also for other applications such as human gait analysis. PMID- 16045219 TI - Rotationally invariant pattern recognition by use of linear and nonlinear cascaded filters. AB - We discuss the merits of using single-layer (linear and nonlinear) and multiple layer (nonlinear) filters for rotationally invariant and noise-tolerant pattern recognition. The capability of each approach is considered with reference to a two-class, rotation-invariant, character recognition problem. The minimum average correlation energy (MACE) filter is a linear filter that is generally accepted to be optimal for detecting signals that are free from noise. Here it is found that an optimized MACE filter cannot differentiate between the characters E and F in a rotation-invariant manner. We have found, however, that this task is possible when a single optimized linear filter is used to achieve the required response when a nonlinear threshold function is included after the filter. We show that this structure can be cascaded to form a multiple-layer, cascaded filter and that the capability of such a system is enhanced by its increased noise tolerance in the character recognition problem. Finally, we show the capability of a two-layer cascade as a means to detect different species of bacteria in images obtained from a phase-contrast microscope. PMID- 16045220 TI - Blurred image recovery by a phase-processing method with tilt correction. AB - We present an improved method that incorporates tilt correction for blurred image recovery. A previously proposed method, which can compensate image blurring due to atmospheric turbulence, was improved to increase recovered image quality by correcting the tilt effect resulting from, e.g., a telescope's tracking jitter. Real-image analysis based on data from a 1.5 m diameter telescope with an infrared camera as well as from a computer simulation were performed. The results suggest that using the tilt-correction method yields an image quality that is much better than that obtained without the correction. PMID- 16045221 TI - Spectral characteristics of asphalt road aging and deterioration: implications for remote-sensing applications. AB - We integrate ground spectrometry, imaging spectrometry, and in situ pavement condition surveys for assessment of asphalt road infrastructure. There is strong spectral evidence for asphalt aging and deterioration. Several spectral measures derived from field and image spectra correlate well with pavement quality indicators (e.g., a pavement condition index). However, the potential for mapping is limited by fine spatial resolution requirements (as fine as 0.5 m) and by the spectral confusion between pavement material aging and asphalt mix erosion on the one hand and structural road damages (e.g., cracking) on the other. PMID- 16045222 TI - Unpolarized infrared emissivity with shadow from anisotropic rough sea surfaces with non-Gaussian statistics. AB - The emissivity of two-dimensional anisotropic rough sea surfaces with non Gaussian statistics is investigated. The emissivity derivation is of importance for retrieval of the sea-surface temperature or equivalent temperature of a rough sea surface by infrared thermal imaging. The well-known Cox-Munk slope probability-density function, considered non-Gaussian, is used for the emissivity derivation, in which the skewness and the kurtosis (related to the third- and fourth-order statistics, respectively) are included. The shadowing effect, which is significant for grazing angles, is also taken into account. The geometric optics approximation is assumed to be valid, which means that the rough surface is modeled as a collection of facets reflecting locally the light in the specular direction. In addition, multiple reflections are ignored. Numerical results of the emissivity are presented for Gaussian and non-Gaussian statistics, for moderate wind speeds, for near-infrared wavelengths, for emission angles ranging from 0 degrees (nadir) to 90 degrees (horizon), and according to the wind direction. In addition, the emissivity is compared with both measurements and a Monte Carlo ray-tracing method. PMID- 16045223 TI - Analysis of transmission spectra for large ratio of emission-to-absorber linewidths: extension of differential absorption lidar analysis for finite laser linewidths. AB - A simple algorithm is presented for the analysis of transmission spectra provided by a lidar with an emission linewidth that is comparable with or larger than the absorption features of interest. The spreading of line shapes as seen by the lidar precludes use of the classical differential absorption lidar (DIAL) approach. However, it is assumed that, as with the DIAL method, small spectral intervals exist where single absorbers are dominant, and an inversion process for the transmission over such intervals is carried out for the absorber concentration. A second-stage algorithm based on singular-value decomposition is also provided to improve further the concentration estimates. An example situation for use of the algorithms is included wherein the objective is to estimate the concentration of a known trace gas in a composite transmission spectrum in the mid-infrared, where the dominant absorbers are water vapor and methane. PMID- 16045224 TI - Performance of a photothermal detector with turbid liquids. AB - A closed-cell photothermal detector for aqueous analytes has been evaluated at 254 and 678 nm. We used a detector with a water meniscus as a pressure sensor, whose periodic deflection was measured using a low-finesse optical fiber Fabry Perot interferometer. Performance was compared with a commercial diode array spectrometer and found to be similar for absorption measurements in nonturbid samples, but the results were affected up to 60 times less by scattered light. Finally the photothermal cell was converted into an integrating cavity using ceramic inserts, showing freedom from scattering-related errors at 678 nm but a degradation in performance at 254 nm. PMID- 16045225 TI - Effect of encoder--decoder mismatch due to wavelength and time misalignments on the performance of two-dimensional wavelength--time optical code-division multiple access systems. AB - We examine the effects of encoder and decoder mismatch due to wavelength and time chip misalignments on the bit-error rate (BER) performance of two-dimensional (2D) wavelength--time optical code-division multiple access systems. We investigate several instances of misalignment in the desired user encoder and decoder as well as in the interfering user encoders. Our simulation methodology can be used to analyze any type of 2D wavelength--time code family as well as probability distribution for misalignment. For illustration purposes, we consider codes generated by use of the depth-first search algorithm and a Gaussian distribution for the misalignment. Our simulation results show that, in the case of a misalignment in either wavelength or time chip, the variance of the distribution for the misalignment must be below 0.01 for the corresponding degradation in the BER system's performance to be less than 1 order of magnitude compared with that when there is no mismatch between the encoders and decoders. The tolerances become even more strict when misalignments in both wavelength and time chips are considered. Furthermore, our results show that the effect of misalignment in wavelength (time chips) is the same regardless of the number of wavelengths (time chips) used in the codes. PMID- 16045226 TI - Stitch error effect on group-delay ripple for long chirped fiber Bragg gratings. AB - Different sources of stitch errors are identified and simulated in a 200 mm long chirped grating. In each case, the effect on the group-delay ripple and transmission spectrum is investigated. The anticipated response in a typical 10 Gbit/s transmission system is specifically considered. From the simulations, it is clear that information about error source and magnitude can be gained directly from the transmission spectrum. PMID- 16045227 TI - Passively mode-locking Nd:Gd0.5Y0.5VO4 laser with an In0.25Ga0.75As absorber grown at low temperature. AB - We have demonstrated stable self-starting passive mode locking in a diode-end pumped Nd:Gd0.5Y0.5VO4 laser by using an In0.25Ga0.75As absorber grown at low temperature (LT In0.25Ga0.75As absorber). An In0.25Ga0.75As single-quantum-well absorber, which was grown directly on the GaAs buffer by use of the metal-organic chemical-vapor deposition technique, acts simultaneously as a passive mode locking device and as an output coupler. Continuous-wave mode-locked pulses were obtained at 1063.5 nm. We achieved a pulse duration of 2.6 ps and an average output power of 2.15 W at a repetition rate of 96.4 MHz. PMID- 16045228 TI - Efficiency optimization for a diode-pumped Nd:YAG ceramic slab laser. AB - With the help of photometric calculations based on ray-tracing algorithms, we have optimized the efficiency of the optical pumping of a Nd:YAG ceramic slab laser. The slab pumping is performed by means of two horizontal diode laser array stacks. The use of two small reflecting walls allows the sort of duct coupling that is capable of significantly improving the performance of the system. Our first experiments with a simple direct coupling provided a maximum extraction of slightly more than 160 W at a 20% slope efficiency level. The use of the optimized short duct coupling leads us to the extraction of 350 W with a slope efficiency of 51%, making use of the same diode arrays. The laser design is suitable for the construction of cw sources with a power output above 1 kW. PMID- 16045229 TI - Variable optical attenuator with a polymer-stabilized dual-frequency liquid crystal. AB - A transmission-type variable optical attenuator (VOA) based on a polymer stabilized dual-frequency liquid crystal (PSDFLC) is demonstrated at gamma = 1.55 microm. The VOA is highly transparent in the voltage-off state but scatters light in the voltage-on state. By using a birefringent beam displacer incorporated with half-wave plates, we can obtain a VOA that is polarization independent and that exhibits a 31 dB dynamic range. The polymer networks and dual-frequency effect together reduce the response time (rise + decay) of a 16 microm PSDFLC cell to 30 ms at room temperature and at a voltage of 24 Vrms. PMID- 16045230 TI - Gap modes of one-dimensional photonic crystal surface waves. AB - The finite-difference time-domain method is employed for the analysis of coupling of the surface modes of two truncated one-dimensional photonic crystals separated by a gap. The wave vector, field distributions, and existence conditions of the coupled surface modes are investigated. The wave vector of symmetric gap modes increases with decreasing gap width, while that of antisymmetric modes decreases exactly opposite of the situation for surface plasmons on metallic half-spaces separated by a dielectric gap. Photonic crystal gap modes could easily and effectively be used as nondissipating gap-mode waveguides. PMID- 16045231 TI - Coating uniformity improvement for a dense-wavelength-division-multiplexing filter by use of the etching effect. AB - The uniformity of optical narrow-bandpass filters for dense wavelength division multiplexing (DWDM) has been improved by control of the coating parameters of electron guns and the ion source. The optical film was deposited by the electron gun and was etched by the ion source during the ion-assisted deposition process. The uniformity of the coating of a 100 GHz DWDM filter is better than +/- 0.003% over a circular area of 50 mm in diameter when such an etching process is used. PMID- 16045232 TI - Cross-correlation frequency-resolved optical gating for studying ultrashort-pulse nonlinear dynamics in arbitrary fibers. AB - We describe a cross-correlation frequency-resolved optical-gating system specifically designed for studying nonlinear pulse-propagation dynamics in fibers of arbitrary length at telecommunication wavelengths. The formation of optical solitons and the appearance of temporal phase slips are observed in 100 m of fiber. The wide phase-matching bandwidth and high sensitivity of this system allow us to visualize femtosecond-pulse evolution in a range of linear and nonlinear propagation regimes. PMID- 16045233 TI - ONS leads the way in oncology nursing-sensitive outcomes research. PMID- 16045235 TI - Recent legislation changes the economics of cancer care. PMID- 16045236 TI - A 14-gene region of rat chromosome 8 in SHR-derived polydactylous congenic substrain affects muscle-specific insulin resistance, dyslipidaemia and visceral adiposity. AB - The SHR and the PD/Cub are two established rodent models of human metabolic syndrome. Introgression of a ca 30 cM region of rat chromosome 8 from PD/Cub onto the genetic background of SHR was previously shown to influence several of the metabolic syndrome-related traits along with causing the PLS in the SHR-Lx congenic strain. In the process of identification of the causative alleles, we have produced several congenic sublines. The differential segment of SHR-Lx PD5 congenic substrain [SHR.PD(D8Rat42-D8Arb23)/Cub] spans approximately 1.4 Mb encompassing only 14 genes. When comparing the metabolic, morphometric and gene expression profiles of the SHR-Lx PD5 vs. SHR, the polydactyly and several distinct metabolic features observed in the original SHR-Lx congenic were still manifested, suggesting that the responsible genes were "trapped" within the relatively short differential segment of PD/Cub origin in SHR-Lx PD5. Particularly, the SHR-Lx PD5 displayed substantial reduction of insulin sensitivity confined to skeletal muscle. Among the candidate genes, the promyelocytic leukaemia zinc-finger Plzf (Zbtb16) transcription repressor is most likely responsible for the Lx mutation resulting in PLS and could also be involved in the alteration of metabolic pathways. The sequence analysis of the Plzf gene revealed a SNP leading to a threonine to serine substitution in SHR at aminoacid position 208 (T208S). In summary, we have isolated a 1.4 Mb genomic region syntenic to human chromosome 11q23, which, apart from causing polydactyly luxate syndrome (PLS), affects total body weight, adiposity, lipid profile, insulin sensitivity of skeletal muscle and related gene expression as shown in the SHR-Lx PD5 congenic substrain. PMID- 16045237 TI - Cytotoxicity and apoptotic effects of microcystin-LR and anatoxin-a in mouse lymphocytes. AB - There is an increasing amount of knowledge on the cytotoxic properties of cyanotoxins, but relatively little is known regarding their fine specificity and mechanisms of action. In this study, we investigated the influence of microcystin LR and AnTx-a on mouse B- and T-lymphocyte subpopulations in vitro. Cyanotoxins significantly decreased the cell viability after 4 and 24 h, compared to the untreated control. After 24 h exposure to microcystin-LR and anatoxin-a, the viability of splenocytes dropped to 23% and 57%, respectively. Our data demonstrate that microcystin-LR induced apoptosis specifically in mouse B cells, probably via the B-cell antigen receptor and mitochondrial pathway, while the T cells were not affected. AnTx-a showed cytotoxic effects on both lymphocyte subpopulations, but the effects were driven by mechanisms different from apoptosis. These findings demonstrate that the cyanotoxins could cause cytotoxic alterations in a variety of cell types different from the major targets, operating via distinct mechanisms. PMID- 16045238 TI - Evaluating the karyotypic diversity in species of Hyla (Anura; Hylidae) with 2n = 30 chromosomes based on the analysis of ten species. AB - Ten species of Hyla with 2n = 30 from Brazilian fauna were analysed cytogenetically. Hyla minuta is the unique presenting all bi-armed metacentric or submetacentric chromosomes in the karyotype, therefore, with the highest FN = 60. The remaining species have a variable number of uni-armed telocentric or subtelocentric chromosomes: H. cruzi, H. elianeae, and H. rubicundula with three pairs (FN = 54), H. berthalutzae, H. elegans, H. microps, and H. nana with four pairs (FN = 52), and H. nahdereri and H. sanborni with five pairs (FN = 50). The uni-armed elements are among pairs 5, 6, 7, 11, 14, and 15, which also appeared with metacentric or submetacentric morphology. The remaining chromosome pairs 1, 2, 3, 4, 8, 9, 10, 12, and 13 were never found to be telocentric or subtelocentric. AgNOR patterns are species-specific, the majority of the species exhibiting a single pair with AgNORs, with the exception of H. elegans and H. nana with more than one chromosome pair bearing this cytological marker. C banding was obtained in H. berthalutzae, H. cruzi, H. elegans, H. elianeae, H. microps, H. minuta, H. nahdereri, and H. nana, which showed positively stained centromeric heterochromatin. Our analysis confirms the great karyotypic diversity in the species of Hyla with 2n = 30, with no species sharing identical karyotypes. PMID- 16045239 TI - Are the T/C polymorphism of the CYP17 gene and the tetranucleotide repeat (TTTA) polymorphism of the CYP19 gene genetic markers for premature coronary artery disease in Caucasians? AB - Gender differences in CAD have been clearly documented, and sex hormones have been recognized to influence the risk of CAD. The cytochrome P450c17alpha gene (CYP17) and the CYP19 gene influence concentrations of sex hormones. In this cross-sectional association study we tested the hypothesis whether the T/C polymorphism of the CYP17 gene and the tetranucleotide repeat (TTTA) polymorphism of the CYP19 gene are genetic markers for CAD in Caucasians. The TT genotype of the CYP17 gene polymorphism was not associated with premature CAD in men and women combined (OR 0.9; 95% CI = 0.6-1.4; P = 0.7), in men only (OR 1; 95% CI = 0.6-1.8; P = 0.7), and in women only (OR 0.8; 95% CI = 0.5-1.4; P = 0.4). The tetranucleotide repeat (TTTA) CYP19 gene polymorphism was not associated with premature CAD. Moreover, the genotypes containing the longer alleles (A6 or A7) were not associated with a lower incidence of CAD, and the genotypes containing the shorter alleles (A1 or A2) were not over-represented in the CAD patients. We may conclude that in Caucasian subjects neither the T/C CYP17 gene polymorphism nor the tetranucleotide repeat (TTTA) polymorphism of the CYP19 gene contributes to the genetic susceptibility to CAD, therefore they may not be used as genetic markers for CAD risk assessment. PMID- 16045240 TI - The effect of gemcitabine in the treatment of rejection in experimental small intestine transplantation. AB - The aim of our study was to test the immunosuppressive effect of gemcitabine in monotherapy following heterotopic SBT in the rat. The BN and LEW rats were used as donors and recipients, respectively. Recipients were divided into 4 groups- group A without immunosuppression, group B treated with a therapeutic dose of tacrolimus, groups C and D treated with various doses of gemcitabine (100 and 150 microg/kg/day). Immunosuppression was administered once a day for 7 days after SBT, when the animals were sacrificed and a histological examination of grafts was performed. Only in group B no signs of acute rejection were seen. Significant differences (P < 0.01) were noted only between group B versus groups A, C, and D. No significant differences were demonstrated between groups A versus groups C, D and between group C versus group D. Monotherapy by gemcitabine (when administered at given doses) was not shown to be effective in preventing acute rejection in a rat model of heterotopic SBT. PMID- 16045241 TI - Can practical nursing ethics be taught? PMID- 16045242 TI - Struggling with the fragility of life: a relational-narrative approach to ethics in palliative nursing. AB - In nursing ethics the role of narratives and dialogue has become more prominent in recent years. The purpose of this article is to illuminate a relational narrative approach to ethics in the context of palliative nursing. The case study presented concerns a difficult relationship between oncology nurses and a husband whose wife was hospitalized with cancer. The husband's narrative is an expression of depression, social isolation and the loss of hope. He found no meaning in the process of dying and death. The oncology nurses were not able to recognize his emotional and existential problems. A narrative perspective inspired by relational ethics indicates that participants may develop a relational narrative that seeks good for all involved in a situation. In palliative nursing this entails open communication about the fragility of life and approaching death. In relational narratives, answers to these ethical dilemmas are co-authored, contingent and contextual. PMID- 16045243 TI - Advance directives: the New Zealand context. AB - Advance directives convey consumers' wishes about accepting or refusing future treatment if they become incompetent. They are designed to communicate a competent consumer's perspective regarding the preferred treatment, should the consumer later become incompetent. There are associated ethical issues for health practitioners and this article considers the features that are relevant to nurses. In New Zealand, consumers have a legal right to use an advance directive that is not limited to life-prolonging care and includes general health procedures. Concerns may arise regarding a consumer's competence and the document's validity. Nurses need to understand their legal and professional obligations to comply with an advance directive. What role does a nurse play and what questions arise for a nurse when advance directives are discussed with consumers? This article considers the cultural dimensions, legal boundaries, consumers' and providers' perspectives, and the medical and nursing positions in New Zealand. PMID- 16045244 TI - The ethics of self-management preparation for chronic illness. AB - While nearly all patients with a chronic disease must self-manage their condition to some extent, preparation for these responsibilities is infrequently assured in the USA. The result can be significant harm and the undermining of a patient's ability to take advantage of life opportunities and be productive. Agreeing to care for a patient involves a moral responsibility to see that she or he receives the essential elements of care, including the ability to manage the disease on a daily basis. The research base for the efficacy of self-management and for how patients can be prepared to assume it is sufficiently strong that health care professionals must advocate for its inclusion in the routine evidence-based care of individuals with chronic disease. Because patient education is central to nursing's philosophy and practice, the profession should play a major role in removing structural barriers to self-management preparation and assuring its provision to a high standard of quality. PMID- 16045245 TI - Justifying coercion. AB - A grounded theory study of psychiatric nurses' experiences of administering medication to involuntary psychiatric patients revealed a basic social process of justifying coercion. Although the 17 nurses interviewed all reported success at avoiding the use of coercion, each had an individual approach to using the nurse patient relationship to do this. However, all the nurses used the same process to reconcile themselves to using coercion when it became necessary. This has three stages: assessment of need; negotiation; and justifying and taking coercive action. Two critical junctures--decision to engage and impasse - determine the progression from one stage to the next. The process of justifying coercion allows a nurse to engage in behavior generally disapproved of while retaining a self image of a 'good' nurse. PMID- 16045246 TI - Nurse moral distress and ethical work environment. AB - This study examined the relationship between moral distress intensity, moral distress frequency and the ethical work environment, and explored the relationship of demographic characteristics to moral distress intensity and frequency. A group of 106 nurses from two large medical centers reported moderate levels of moral distress intensity, low levels of moral distress frequency, and a moderately positive ethical work environment. Moral distress intensity and ethical work environment were correlated with moral distress frequency. Age was negatively correlated with moral distress intensity, whereas being African American was related to higher levels of moral distress intensity. The ethical work environment predicted moral distress intensity. These results reveal a difference between moral distress intensity and frequency and the importance of the environment to moral distress intensity. PMID- 16045247 TI - Emotional boundary work in advanced fertility nursing roles. AB - In this article we examine the nature of intimacy and knowing in the nurse patient relationship in the context of advanced nursing roles in fertility care. We suggest that psychoanalytical approaches to emotions may contribute to an increased understanding of how emotions are managed in advanced nursing roles. These roles include nurses undertaking tasks that were formerly performed by doctors. Rather than limiting the potential for intimacy between nurses and fertility patients, we argue that such roles allow nurses to provide increased continuity of care. This facilitates the management of emotions where a feeling of closeness is created while at the same time maintaining a distance or safe boundary with which both nurses and patients are comfortable. We argue that this distanced or 'bounded' relationship can be understood as a defence against the anxiety of emotions raised in the nurse-fertility patient relationship. PMID- 16045248 TI - Introductory engagement within the perinatal nursing relationship. AB - In this article, the theme of introductory engagement is developed through the conversational interviews and participatory observations I carried out with perinatal nurses and birthing women in the context of a feminist phenomenological methodology. Positioned against the landscape of hierarchical health care practices embedded with power dynamics and disembodied practices, this research explored the ways in which perinatal nurses related to birthing women in the context of relational care. The focus of attention in this article is to describe the theme of introductory engagement by way of a storied phenomenological text. PMID- 16045249 TI - Country profile: Canada's health care system. PMID- 16045250 TI - Infants can use distributional cues to form syntactic categories. AB - Nearly all theories of language development emphasize the importance of distributional cues for segregating words and phrases into syntactic categories like noun, feminine or verb phrase. However, questions concerning whether such cues can be used to the exclusion of referential cues have been debated. Using the headturn preference procedure, American children aged 1;5 were briefly familiarized with a partial Russian gender paradigm, with a subset of the paradigm members withheld. During test, infants listened on alternate trials to previously withheld grammatical items and ungrammatical items with incorrect gender markings on previously heard stems. Across three experiments, infants discriminated new grammatical from ungrammatical items, but like adults in previous studies, were only able to do so when a subset of familiarization items was double marked for gender category. The results suggest that learners can use distributional cues to category structure, to the exclusion of referential cues, from relatively early in the language learning process. PMID- 16045251 TI - Testing the agreement/tense omission model: why the data on children's use of non nominative 3psg subjects count against the ATOM. AB - One of the most influential recent accounts of pronoun case-marking errors in young children's speech is Schutze & Wexler's (1996) Agreement/Tense Omission Model (ATOM). The ATOM predicts that the rate of agreeing verbs with non nominative subjects will be so low that such errors can be reasonably disregarded as noise in the data. The present study tests this prediction on data from 12 children between the ages of 1; 8.22 and 3 ; 0.10. This is done, first, by identifying children who produced a reasonably large number of non-nominative 3psg subjects; second, by estimating the expected rate of agreeing verbs with masculine and feminine non-nominative subjects in these children's speech; and, third, by examining the actual rate at which agreeing verb forms occurred with non-nominative subjects in those areas of the data in which the expected error rate was significantly greater than 10%. The results show, first, that only three of the children produced enough non-nominative subjects to allow a reasonable test of the ATOM to be made; second, that for all three of these children, the only area of the data in which the expected frequency of agreeing verbs with non nominative subjects was significantly greater than 10% was their use of feminine case-marked subjects; and third, that for all three of these children, the rate of agreeing verbs with nonnominative feminine subjects was over 30%. These results raise serious doubts about the claim that children's use of non nominative subjects can be explained in terms of AGR optionality, and suggest the need for a model of pronoun case-marking error that can explain why some children produce agreeing verb forms with non-nominative subjects as often as they do. PMID- 16045252 TI - On learning to draw the distinction between physical and metaphorical motion: is metaphor an early emerging cognitive and linguistic capacity? AB - Situated within the framework of the conceptual metaphor theory (Lakoff & Johnson, 1999), this study investigated young children's understanding of metaphorical extensions of spatial motion. Metaphor was defined as a conceptual linguistic mapping between a source and a target domain. The study focused on metaphors that are structured by the source domain of motion in space (e.g. time flies by, ideas pass through one's mind, sickness crawls through one's body). The study investigated whether metaphor comprehension varied by the age of the participant, target domain of the metaphorical mapping, and the conventionality of the linguistic form with which the metaphor was conveyed. Data were gathered using a story comprehension task and a semi-structured interview from 60 monolingual Turkish-speaking children, at the mean ages 3;6, 4;5 and 5;5 (20 participants per age group), and 20 adult native speakers of Turkish. The results showed metaphor understanding to be an early emerging cognitive and linguistic capacity. PMID- 16045253 TI - Children's resistance to homonymy: an experimental study of pseudohomonyms. AB - Research in diachronic linguistics has shown that homonyms are often dispreferred in language. This study proposes that this trend is mirrored in the difficulties that children encounter in mapping homonyms. Two experiments are presented in support of this proposition. In Experiment 1, 16 preschool children (mean age = 4;6) are shown to perform quite well on tasks requiring them to assign novel meanings to nonsense words. They perform poorly, however, on tasks requiring them to assign a different, unrelated meaning to a known word. Experiment 2 (N = 18, mean age = 4;5) shows that preschoolers' performance on this task, however, improves when a known word appears in a syntactic frame that is not appropriate for the word (as when a verb appears in a noun syntactic frame), thereby providing a strong indication that a new meaning is appropriate. PMID- 16045254 TI - Acquisition of English comparative adjectives. AB - Two experiments investigated the acquisition of English comparative adjective forms, Adj + er and more Adj. In Experiment 1, 72 children, four- and seven-years old, indicated their preferences for the synthetic or periphrastic comparative form for 16 adjectives in a forced-choice judgement task; their responses were compared to those of a group of adults (Graziano-King, 2003). In Experiment 2, a group of 29 children, ranging in age from 5;1 to 10;9, and a group of 11 adults performed a forced-choice judgement task, similar to that of Experiment 1, and an elicited production task, responding to the same 32 adjectives for both tasks. The two studies together support an acquisition trajectory of three stages. In the first stage, children show no preference for either form of the comparative; in the second, they adopt a suffixation rule; and in the third, they abandon the general rule and become conservative learners, eventually reaching the adult target. PMID- 16045255 TI - The strength of children's knowledge of the role of root morphemes in the spelling of derived words. AB - The spelling of words in English is governed in part by the morphemes that make them up. This study examines the strength of children's knowledge of the role of root morphemes in spelling, specifically focusing on whether it can withstand interference by phonological changes. A total of 75 children between seven and nine years of age were given the first parts of two-morpheme transparent and opaque derived words (e.g. musical and objection) and one-morpheme control words (e.g. metal and portion). Children used the clues to a significant extent with the two-, but not the one-morpheme words. This effect was consistent across age groups and across both types of derived words. This is evidence that children's understanding of the link between morphemes and spelling is not impeded by changes in the sound of morphemes. These findings have substantial implications for models of spelling development and for educational practice. PMID- 16045256 TI - Notes on Ingram's whole-word measures for phonological development. AB - In this note we discuss pMLU, a whole-word measure for phonological development that was proposed by Ingram (2002). Ingram's rules for calculating pMLU are analysed and we point at the crucial role of the level of transcription for making pMLU measurements comparable over different corpora. The main aim of the paper is an assessment of the reliability and the validity of pMLU. The assessment is accomplished using a computational tool for measuring pMLU on two large Dutch CHILDES corpora. We propose minimal sample sizes for reliable measurements relative to the stage of phonological development. PMID- 16045257 TI - The role of prediction in construction-learning. AB - It is well-established that (non-linguistic) categorization is driven by a functional demand of prediction. We suggest that prediction likewise may well play a role in motivating the learning of semantic generalizations about argument structure constructions. We report corpora statistics that indicate that the argument frame or construction has roughly equivalent cue validity as a predictor of overall sentence meaning as the morphological form of the verb, and has greater category validity. That is, the construction is at least as reliable and more available than the verb. Moreover, given the fact that many verbs have quite low cue validity in isolation, attention to the contribution of the construction is essential. PMID- 16045258 TI - Asynchrony in the cognitive and lexical development of young children with Williams syndrome. AB - The present study investigates whether five-to-six-year-old children with Williams syndrome (N = 8) can form new object categories based on naming information alone, and compares them with five groups of typically developing children aged 2;0 to 6;0 (N = 34 children). Children were presented with triads of dissimilar objects; all objects in a triad were labelled, two of them with the same pseudoname. Name-based categorization was evaluated through object selection. Performance was above chance level for all groups. Performance reached a ceiling at about 4;0 for the typically developing children. For the children with Williams Syndrome, performance remained below chronological age level. The present results are discussed in light of previous findings of a failure to perform name-based categorization in younger children with Williams syndrome and the persistent asynchrony between cognitive and lexical development in this disorder. PMID- 16045259 TI - The use of anaphoric pronouns by French children in narrative: evidence from constrained text production. AB - This paper describes the acquisition of the 3rd person pronoun 'il/elle' (he, she, it) in seven to twelve-year-old French children (N = 58), in written production. An experiment was conducted to examine the relationship between the use of this anaphoric pronoun and the accessibility of the memory-trace of the corresponding referent in the texts. Referential accessibility in short texts was varied according to three factors: referential distance, thematization of the agent role (first sentence subject), and discourse focus. We found that the children were sensitive to the distance factor as early as 7;0, i.e. they used fewer personal pronouns when the referential distance increased. However, children of different ages differed in their weighting of the discourse focus factor and the thematization factor: the seven-year-olds (N = 18) and the eleven year-olds (N = 20) were sensitive to variation of the discourse focus but not the thematization factor, while for the nine-year-olds (N = 20) it was the reverse. The main results suggest: (a) when seven and nine-year-olds use the pronoun 'il/elle', they do not comply with the constraints associated with the accessibility of the memory-trace of the referent; (b) memory constraints have an effect from the age of 7;0, but only when the discourse focus is maintained. It was concluded that the discourse management of the French personal pronoun 'il/elle' is not totally mastered at 11;0: children cannot operationally integrate the whole array of constraints implied in anaphoric management. PMID- 16045260 TI - If you prescribe will they dispense? PMID- 16045261 TI - Fired for protecting patients? PMID- 16045262 TI - 10 more ways to work smarter. PMID- 16045263 TI - Another way to get sued. PMID- 16045264 TI - When the news is bad. PMID- 16045265 TI - Why I deliver bad news myself. PMID- 16045266 TI - Taking stock. A colleague's sudden death prompted the author to step back and think about his life. PMID- 16045267 TI - He's not my patient, he's my father. PMID- 16045268 TI - Financial survey. Is your net worth keeping pace? PMID- 16045270 TI - We don't have it so bad. PMID- 16045269 TI - Liability for house calls. PMID- 16045271 TI - Donald Watkins--the man who saved Richard Scrushy. PMID- 16045272 TI - Can GM slash worker benefits? PMID- 16045273 TI - Wrestling chiropractor. PMID- 16045274 TI - A hierarchical clustering approach for large compound libraries. AB - A modified version of the k-means clustering algorithm was developed that is able to analyze large compound libraries. A distance threshold determined by plotting the sum of radii of leaf clusters was used as a termination criterion for the clustering process. Hierarchical trees were constructed that can be used to obtain an overview of the data distribution and inherent cluster structure. The approach is also applicable to ligand-based virtual screening with the aim to generate preferred screening collections or focused compound libraries. Retrospective analysis of two activity classes was performed: inhibitors of caspase 1 [interleukin 1 (IL1) cleaving enzyme, ICE] and glucocorticoid receptor ligands. The MDL Drug Data Report (MDDR) and Collection of Bioactive Reference Analogues (COBRA) databases served as the compound pool, for which binary trees were produced. Molecules were encoded by all Molecular Operating Environment 2D descriptors and topological pharmacophore atom types. Individual clusters were assessed for their purity and enrichment of actives belonging to the two ligand classes. Significant enrichment was observed in individual branches of the cluster tree. After clustering a combined database of MDDR, COBRA, and the SPECS catalog, it was possible to retrieve MDDR ICE inhibitors with new scaffolds using COBRA ICE inhibitors as seeds. A Java implementation of the clustering method is available via the Internet (http://www.modlab.de). PMID- 16045275 TI - ThermoData Engine (TDE): software implementation of the dynamic data evaluation concept. AB - The first full-scale software implementation of the dynamic data evaluation concept {ThermoData Engine (TDE)} is described for thermophysical property data. This concept requires the development of large electronic databases capable of storing essentially all experimental data known to date with detailed descriptions of relevant metadata and uncertainties. The combination of these electronic databases with expert-system software, designed to automatically generate recommended data based on available experimental data, leads to the ability to produce critically evaluated data dynamically or 'to order'. Six major design tasks are described with emphasis on the software architecture for automated critical evaluation including dynamic selection and application of prediction methods and enforcement of thermodynamic consistency. The direction of future enhancements is discussed. PMID- 16045276 TI - A stepwise approach for defining the applicability domain of SAR and QSAR models. AB - A stepwise approach for determining the model applicability domain is proposed. Four stages are applied to account for the diversity and complexity of the current SAR/QSAR models, reflecting their mechanistic rationality (including metabolic activation of chemicals) and transparency. General parametric requirements are imposed in the first stage, specifying in the domain only those chemicals that fall in the range of variation of the physicochemical properties of the chemicals in the training set. The second stage defines the structural similarity between chemicals that are correctly predicted by the model. The structural neighborhood of atom-centered fragments is used to determine this similarity. The third stage in defining the domain is based on a mechanistic understanding of the modeled phenomenon. Here, the model domain combines the reliability of specific reactive groups hypothesized to cause the effect and the domain of explanatory variables determining the parametric requirements in order for functional groups to elicit their reactivity. Finally, the reliability of simulated metabolism (metabolites, pathways, and maps) is taken into account in assessing the reliability of predictions, if metabolic activation of chemicals is a part of the (Q)SAR model. Some of the stages of the proposed approach for defining the model domain can be eliminated depending on the availability and quality of the experimental data used to derive the model, the specificity of (Q)SARs, and the goals of their ultimate application. The performance of the proposed definition of the model domain is tested using several examples of (Q)SARs that have been externally validated, including models for predicting acute toxicity, skin sensitization, and biodegradation. The results clearly showed that credibility in predictions of QSAR models for chemicals belonging to their domain is much higher than for chemicals outside this domain. PMID- 16045277 TI - Stability and properties of polyhelicenes and annelated fused-ring carbon helices: models toward helical graphites. AB - The geometrical structures and properties of conjugated polyhelicenes and annelated fused-ring carbon helices with analogous frameworks were theoretically studied at the HF/6-31G and B3LYP/6-31G levels. These studies focused on the stability of the fused-ring structures with special emphasis on the helical geometrical arrangements. To elucidate bonding patterns, the orbitals, electron density contours, and the electrostatic potential of these helical compounds were analyzed. The structure of fused polynaphthalenes arranged in a helical spiral can be regarded as part of a locally helical graphite lattice that is expected to give rise to special electronic properties along the helically layered conjugated single sheet that can be regarded as a single extended pi system but also involving local layer-to-layer pi-pi interactions that are typical in ordinary graphite. This dual feature might lead to novel materials. PMID- 16045278 TI - A new rapid and effective chemistry space filter in recognizing a druglike database. AB - To develop a new chemistry space filter with high efficiency and accuracy, an analysis on distributions of as many as 50 structural and physicochemical properties was carried out on both druglike and nondruglike databases, viz. MACCS II Drug Data Report (MDDR), Comprehensive Medicinal Chemistry (CMC), and Available Chemicals Directory (ACD). Based on the analysis results, a chemistry space filter was developed that can effectively discriminate a druglike database from a nondruglike database. The filter is composed of two descriptors: one is a molecular saturation related descriptor, and the other is associated with the proportion of heteroatoms in a molecule. Both are molecular size independent. Therefore, the profiles of a druglike database could be characterized as proper molecular saturation and proper percentage of heteroatoms, revealing direct indices for designing and optimizing combinatorial libraries. The application of the new filter on the Chinese Natural Product Database (CNPD) suggested that CNPD is, as expected, a potential druglike database, testifying that the new filter is reliable. Therefore, this newly developed chemistry space filter should be a potent tool for identifying druglike molecules, thus, it would have potential applications in the research of combinatorial library design and virtual high throughput screening using computational approaches for drug discovery. PMID- 16045279 TI - A searching and reporting system for relational databases using a graph-based metadata representation. AB - Relational databases are the current standard for storing and retrieving data in the pharmaceutical and biotech industries. However, retrieving data from a relational database requires specialized knowledge of the database schema and of the SQL query language. At Anadys, we have developed an easy-to-use system for searching and reporting data in a relational database to support our drug discovery project teams. This system is fast and flexible and allows users to access all data without having to write SQL queries. This paper presents the hierarchical, graph-based metadata representation and SQL-construction methods that, together, are the basis of this system's capabilities. PMID- 16045280 TI - Posetic quantitative superstructure/activity relationships (QSSARs) for chlorobenzenes. AB - As a result of the widespread industrial use of polychlorinated hydrocarbons, they have accumulated in nearly all types of environmental compartments, especially in aquatic systems. Particularly, chloroaromatics are among the most undesirable industrial effluents because of their persistence and toxicity. To predict chlorobenzene (CB) toxicities, we make use of a novel scheme that looks beyond simple molecular structure to the manner in which such a structure embeds in an overall reaction network. Thence, a resultant modeling gives a quantitative superstructure/activity relationship (QSSAR) with the (chloro-substitution) reaction network viewed mathematically as a partially ordered set (or poset). Different numerical fittings to the overall poset lead to different QSSAR models, of which we investigate three: average poset, cluster expansion, and splinoid poset QSSAR models for the CBs' toxicities against various species (Poecilia reticulata,Pimephales promelas, Daphnia magna, Rana japonica, etc). Excellent results are obtained for all QSSAR toxicity models. On the basis of the poset reaction diagram, all three of these QSSAR models reflect, in distinct ways, the topology of the network that describes the interconversion of chemical species. Although in the majority of investigated datasets all poset QSSAR models give very good predictions, in some cases, they complement each other. These differences show that more reliable predictions can be obtained by using a consensus prediction that combines data from the three posetic models. PMID- 16045281 TI - Adaptive multicomponent analysis by genetic algorithms. AB - The applicability of genetic algorithms for solving multicomponent analyses is systematically examined. As a genetic algorithm (GA), the basic proposal of Goldberg is implemented in a straightforward manner to simulate multicomponent analyses in analogy to the well-established UV-vis or IR methods, especially multicomponent regression. The main focus of the study is to investigate the behavior of the genetic algorithm in order to compare it with the well-known behavior of multicomponent regression. A remarkable difference between the two methods is that the genetic algorithm method does not need any calibration procedure because of its pure searching characteristic. As important features of multicomponent systems, the degree of signal overlap (selectivity), the behavior of systems with known and unknown component numbers and qualities, and linear as well as nonlinear relationships between the analytical signal and concentration are varied within the simulations. According to multicomponent regression, recovering concentrations by a genetic algorithm is of limited applicability with the exception of systems at a low degree of signal overlap. On the other hand, the recovery of a probe spectrum in the analytical process always gives satisfactory results independent of the features of the probe system. The genetic algorithm obviously shows autoadaptive behavior in probe spectrum recovery. The quality and quantity of the resulting components may dramatically differ from the given probe, although the resulting spectrum is nearly the same. In such cases, the resulting component mixture can be interpreted as an imitation of the probe. As well probe spectra, theoretically designed spectra can also be autoadapted by genetic algorithms. The only limitation is that the desired spectrum must, of course, be incorporated into the search space defined by the involved components. Furthermore, a spectral signal is only one single property of a chemical compound or mixture. Because of the nonlinear search characteristic of genetic algorithms, any other chemical or physical property can also be treated as a desired property. Therefore, the conclusion of the study is well-founded that an old challenge of applied chemistry, namely, the development of new chemical products with desired properties, seems to be reachable under the control of genetic algorithms. PMID- 16045282 TI - "In-house likeness": comparison of large compound collections using artificial neural networks. AB - Binary classification models able to discriminate between data sets of compounds are useful tools in a range of applications from compound acquisition to library design. In this paper we investigate the ability of artificial neural networks to discriminate between compound collections from various sources aiming at developing an "in-house likeness" scoring scheme (i.e. in-house vs external compounds) for compound acquisition. Our analysis shows atom-type based Ghose Crippen fingerprints in combination with artificial neural networks to be an efficient way to construct such filters. A simple measure of the chemical overlap between different compound collections can be derived using the output scores from the neural net models. PMID- 16045283 TI - Improving the quantification of highly overlapping chromatographic peaks by using product unit neural networks modeled by an evolutionary algorithm. AB - This work investigates the ability of multiplicative (on the basis of product units) and sigmoidal neural models built by an evolutionary algorithm to quantify highly overlapping chromatographic peaks. To test this approach, two N methylcarbamate pesticides, carbofuran and propoxur, were quantified using a classic peroxyoxalate chemiluminescence reaction as a detection system for chromatographic analysis. The four-parameter Weibull curve associated with the profile of the chromatographic peak estimated by the Levenberg-Marquardt method was used as input data for both models. Straightforward network topologies (one output) allowed the analytes to be quantified with great accuracy and precision. Product unit neural networks provided better information ability, smaller network architectures, and more robust models (smaller standard deviation). The reduced dimensions of the selected models enabled the derivation of simple quantification equations to transform the input variables into the output variable. These equations can be more easily interpreted from a chemical point of view than those provided by sigmoidal neural networks, and the effect of both analytes on the characteristics of chromatographic bands, namely profile, dispersion, peak height, and residence time, can be readily established. PMID- 16045284 TI - Genetic programming for the induction of decision trees to model ecotoxicity data. AB - Automatic induction of decision trees and production rules from data to develop structure-activity models for toxicity prediction has recently received much attention, and the majority of methodologies reported in the literature are based upon recursive partitioning employing greedy searches to choose the best splitting attribute and value at each node. These approaches can be successful; however, the greedy search will necessarily miss regions of the search space. Recent literature has demonstrated the applicability of genetic programming to decision tree induction to overcome this problem. This paper presents a variant of this novel approach, using fewer mutation options and a simpler fitness function, demonstrating its utility in inducing decision trees for ecotoxicity data, via a case study of two data sets giving improved accuracy and generalization ability over a popular decision tree inducer. PMID- 16045285 TI - A general treatment of solubility. 3. Principal component analysis (PCA) of the solubilities of diverse solutes in diverse solvents. AB - A phenomenological study of solubility has been conducted using a combination of quantitative structure-property relationship (QSPR) and principal component analysis (PCA). A solubility database of 4540 experimental data points was used that utilized available experimental data into a matrix of 154 solvents times 397 solutes. Methodology in which QSPR and PCA are combined was developed to predict the missing values and to fill the data matrix. PCA on the resulting filled matrix, where solutes are observations and solvents are variables, shows 92.55% of coverage with three principal components. The corresponding transposed matrix, in which solvents are observations and solutes are variables, showed 62.96% of coverage with four principal components. PMID- 16045286 TI - Quantum mechanical structure-activity relationship analyses for skin sensitization. AB - Allergic contact dermatitis (ACD) results in inflammation of the skin due to sensitization of the immunologic system to a particular substance. The sensitization process is limited by the compound's ability to both permeate and react with proteins in the integumentary system. Currently, only in vivo animal tests such as the local lymph node assay (LLNA) are recognized by regulatory authorities for risk assessment of ACD. A quantitative structure-activity relationship has been developed to predict relative potency, which allows for the prediction of relative sensitization potentials. The experimental values used in this study include EC3 values (the concentration at which the stimulation index equals 3) from LLNA tests. The predictions in this model enable categorization of the compounds into three groups on the basis of risk of sensitization and enable screening of candidate molecules using rapid SAM1 semiempirical calculations prior to animal testing. The model may also be used to reduce the number of animals subjected to testing by providing estimated concentrations required for useful data of risk assessment. The effect of averaging available literature values on predictive ability is also investigated. The model includes halogenated compounds, aromatic compounds, alcohols, aldehydes, and ketones. The computational investigation resulted in a two-descriptor model that is consistent with the assumed mechanism for sensitization. PMID- 16045287 TI - Modeling the octanol-water partition coefficients by an optimized molecular connectivity index. AB - A procedure that makes it possible to generate a coherent model for prediction of the octanol-water partition coefficient within the molecular connectivity formalism was put forward. The method is based on the optimization of weights for corresponding skeletal atoms and is similar to the method for calculation of a variable connectivity index proposed by Randic. In contrast to Randic's method, we incorporate in the algorithm the possibility that the contribution of a term describing a carbon-heteroatom bond may be negative. When tested on a set of about 300 structurally diverse organic molecules, our procedure proved to be superior to the standard valence connectivity method. External validation on a smaller set of compounds confirmed the superiority of our procedure with respect to the standard one. Intramolecular interactions, which are operative in more complex compounds, are treated in a similar fashion to that in the Hansch-Leo or Rekker methods, by inclusion of empirical correction factors. PMID- 16045288 TI - Graph kernels for molecular structure-activity relationship analysis with support vector machines. AB - The support vector machine algorithm together with graph kernel functions has recently been introduced to model structure-activity relationships (SAR) of molecules from their 2D structure, without the need for explicit molecular descriptor computation. We propose two extensions to this approach with the double goal to reduce the computational burden associated with the model and to enhance its predictive accuracy: description of the molecules by a Morgan index process and definition of a second-order Markov model for random walks on 2D structures. Experiments on two mutagenicity data sets validate the proposed extensions, making this approach a possible complementary alternative to other modeling strategies. PMID- 16045289 TI - Application of the random forest method in studies of local lymph node assay based skin sensitization data. AB - The random forest and classification tree modeling methods are used to build predictive models of the skin sensitization activity of a chemical. A new two stage backward elimination algorithm for descriptor selection in the random forest method is introduced. The predictive performance of the random forest model was maximized by tuning voting thresholds to reflect the unbalanced size of classification groups in available data. Our results show that random forest with a proposed backward elimination procedure outperforms a single classification tree and the standard random forest method in predicting Local Lymph Node Assay based skin sensitization activity. The proximity measure obtained from the random forest is a natural similarity measure that can be used for clustering of chemicals. Based on this measure, the clustering analysis partitioned the chemicals into several groups sharing similar molecular patterns. The improved random forest method demonstrates the potential for future QSAR studies based on a large number of descriptors or when the number of available data points is limited. PMID- 16045290 TI - Stability studies of transition-metal linkage isomers using quantum mechanical methods. Groups 11 and 12 transition metals. AB - Several hypotheses to elucidate the linkage isomer preference of the thiocyanate (SCN(-)) ion have been offered. For complexes with small coordination numbers (i.e., 1 and 2) and groups 11 (Cu-triad) and 12 (Zn-triad) metals, different levels of theory and a variety of basis sets have been employed to study linkage isomerism. Similar results are obtained for all density functionals tested, pure and hybrid. Overall, good agreement, vis-a-vis experimentally identified linkage isomers, is achieved for ab initio techniques, whereas semiempirical quantum mechanical methods show a bias toward S-ligated isomers. Despite the seeming ease for the a priori prediction of the most stable thiocyanate isomers using acid/base principles, this research highlights the sensitivity of quantitative calculations of transition-metal linkage isomerism to the choice of basis set and electron correlation, particularly with post-Hartree-Fock treatments. PMID- 16045291 TI - Exploring phase-transfer catalysis with molecular dynamics and 3D/4D quantitative structure-selectivity relationships. AB - Quantitative Structure-Selectivity Relationships (QSSR) are developed for a library of 40 phase-transfer asymmetric catalysts, based around quaternary ammonium salts, using Comparative Molecular Field Analysis (CoMFA) and closely related variants. Due to the flexibility of these catalysts, we use molecular dynamics (MD) with an implicit Generalized Born solvent model to explore their conformational space. Comparison with crystal data indicates that relevant conformations are obtained and that, furthermore, the correct biphenyl twist conformation is predicted, as illustrated by the superiority of the resulting model (leave-one-out q(2) = 0.78) compared to a random choice of low-energy conformations for each catalyst (average q(2) = 0.22). We extend this model by incorporating the MD trajectory directly into a 4D QSSR and by Boltzmann weighting the contribution of selected minimized conformations, which we refer to as '3.5D' QSSR. The latter method improves on the predictive ability of the 3D QSSR (leave-one-out q(2) = 0.83), as confirmed by repeated training/test splits. PMID- 16045292 TI - Prediction of cytochrome P450 3A4, 2D6, and 2C9 inhibitors and substrates by using support vector machines. AB - Statistical learning methods have been used in developing filters for predicting inhibitors of two P450 isoenzymes, CYP3A4 and CYP2D6. This work explores the use of different statistical learning methods for predicting inhibitors of these enzymes and an additional P450 enzyme, CYP2C9, and the substrates of the three P450 isoenzymes. Two consensus support vector machine (CSVM) methods, "positive majority" (PM-CSVM) and "positive probability" (PP-CSVM), were used in this work. These methods were first tested for the prediction of inhibitors of CYP3A4 and CYP2D6 by using a significantly higher number of inhibitors and noninhibitors than that used in earlier studies. They were then applied to the prediction of inhibitors of CYP2C9 and substrates of the three enzymes. Both methods predict inhibitors of CYP3A4 and CYP2D6 at a similar level of accuracy as those of earlier studies. For classification of inhibitors of CYP2C9, the best CSVM method gives an accuracy of 88.9% for inhibitors and 96.3% for noninhibitors. The accuracies for classification of substrates and nonsubstrates of CYP3A4, CYP2D6, and CYP2C9 are 98.2 and 90.9%, 96.6 and 94.4%, and 85.7 and 98.8%, respectively. Both CSVM methods are potentially useful as filters for predicting inhibitors and substrates of P450 isoenzymes. These methods generally give better accuracies than single SVM classification systems, and the performance of the PP-CSVM method is slightly better than that of the PM-CSVM method. PMID- 16045293 TI - Improved FlexX docking using FlexS-determined base fragment placement. AB - We report on a novel hybrid FlexX/FlexS docking approach, whereby the base fragment of the test ligand is chosen by FlexS superposition onto a cocrystallized template ligand and then fed into FlexX for the incremental construction of the final solution. The new approach is tested on the diverse 200 protein-ligand complex dataset that has been previously described for FlexX validation. In total, 62.9% of the complexes can be reproduced at rank 1 by our approach, which compares favorably with 46.9% when using FlexX alone. In addition, we report "cross-docking" experiments in which several receptor structures of complexes with identical proteins have been used for docking all cocrystallized ligands of these complexes. The results show that, in almost all cases, the hybrid approach can acceptably dock a ligand into a foreign receptor structure using a different ligand template, can give solutions where FlexX alone fails, and tends to give solutions that are more accurately positioned. PMID- 16045294 TI - Nanoporous carbon allotropes by septupling map operations. AB - Spongy carbon nanostructures, also called schwarzites, have been synthesized. They consist of highly connected covalent networks, periodic in the three dimensions of Euclidean space. The intimate structure of schwarzites has a topology of triply periodic minimal surfaces. They can be tessellated by some geometric operations on maps, including the newly proposed septupling operations. Formulas for calculating the lattice parameters of iteratively transformed maps are presented. Examples are given for both finite/closed cages and infinite/open all-sp2 carbon structures. Strain energy calculations for structures, consisting of thousands of atoms, show that such carbon allotropes are very relaxed and approach to the non-strained graphite sheet. PMID- 16045295 TI - An analytical, variable resolution, complete description of static molecules and their intermolecular binding properties. AB - A fully analytical description of molecular shape, as defined by the shrink-wrap isodensity or solvent-excluded surfaces and local properties related to Coulomb, donor/acceptor, and polarizability (dispersion) interactions is described. The molecular surface and four local properties adequate for describing intermolecular interactions (the molecular electrostatic potential and the local ionization energy, electron affinity, and polarizability) are fitted to spherical harmonic expansions, which provide a compact and information-rich description of the properties of the static molecule. The resolution of the description can be varied from isotropic to near atomistic detail by adjusting the order of the individual spherical-harmonic expansions. Examples are given to illustrate the effect of truncating the different spherical-harmonic approximations. PMID- 16045296 TI - Enhanced virtual screening by combined use of two docking methods: getting the most on a limited budget. AB - Flexible ligand docking is a routine part of a modern structure-based lead discovery process. As of today, there are quite a number of commercial docking programs that can be used to screen large databases (hundreds of thousands to millions of compounds). However, limiting factors such as the number of commercial software licenses needed to perform docking simultaneously on multiple processors ("software cost") and the relatively long time required per molecule to get good results ("quality-to-speed") should be taken into account when planning a large docking run. How can we optimize the efficiency of selecting lead candidates by docking, in respect to the quality of the results, search speed, and software cost? We present a combination of two methods, our "fast-free approximate" in-house docking program and the "slow-costly-accurate" ICM-Dock, as an example of one solution to the problem. Our proposed protocol is illustrated by a series of virtual screening experiments aimed at identifying active compounds in the MDL Drug Data Report database. In more than half of the 20 cases examined, at least several actives per protein target were identified in approximately 24 hours per target. PMID- 16045297 TI - Modified ant colony optimization algorithm for variable selection in QSAR modeling: QSAR studies of cyclooxygenase inhibitors. AB - A new version of an ant colony optimization (ACO) algorithm has been proposed. A modified ACO algorithm is proposed to select variables in QSAR modeling and to predict inhibiting action of some diarylimidazole derivatives on cyclooxygenase (COX) enzyme. As a comparison to this method, the evolution algorithm (EA) was also tested. Experimental results have demonstrated that the modified ACO is a useful tool for variable selection that needs few parameters to be adjusted and converges quickly toward the optimal position. PMID- 16045298 TI - Model carbyne vs ideal and DNA catenanes. AB - The structure and stability of two-component carbyne catenanes, viewed as model compounds for DNA catenanes, have been estimated by molecular mechanics (MM) calculations. The carbyne catenane molecules studied were composed from interwined cyclic molecules constituted solely from carbon atoms bonded by alternating single and triple bonds. The total number of carbon atoms in molecules studied was set to 60, and the complementary sizes of the catenane components varied. The component cycles were entangled by 2, 4, ..., 11 crossings. For the catenanes with the number of crossings equal to six and seven more than one catenane topological type was considered. The calculated MM strain energy of carbyne catenanes was correlated with the averaged crossing number and length-to-diameter ratio of ideal catenanes as well as the electrophoretic mobility and sedimentation coefficient of DNA catenanes. To observe these correlations, for each topological type of carbyne catenane, it was necessary to find the proportion of the sizes of the two catenane components at which the MM strain energy is the lowest. Then, the values of these energies were correlated with characteristics of ideal and real DNA catenanes. The fits are significant and nonlinear. PMID- 16045299 TI - Prediction of UV and ESI-MS signal intensities. AB - All major pharmaceutical companies maintain large collections of compounds that are used either for screening against biological targets or as synthetic precursors. The quality assessment of these compounds is typically done by liquid chromatography combined with mass spectroscopy (LC/MS) and UV purity control. To facilitate the analysis of the analytical data, we have built computational models to predict UV and MS signal intensities under experimental LC/MS conditions. The discriminant partial-least-squares technique was used for classifying compounds into those most likely to yield a MS signal and others where the signal is below the detection limit (94% and 88% correct predictions, respectively). In the case of UV prediction, we compared this statistical linear regression technique to a knowledge-based approach. A combination of both techniques proved to be the most reliable (96/98% correct predictions of UV active/ UV-inactive compounds). Both models have been incorporated into the automated compound integrity profiling at F. Hoffmann-La Roche. PMID- 16045300 TI - Computational and conformational evaluation of FTase alternative substrates: insight into a novel enzyme binding pocket. AB - Protein farnesyltransferase (FTase) is an important anticancer drug target. In an effort to develop isoprenoid diphosphate-based FTase inhibitors, striking variations have been observed in the ability of conservatively modified analogues to bind to the enzyme. For example, 2Z-GGPP is an alternative substrate with high binding affinity, while GGPP is not an alternative substrate. Using the availability of high-resolution FTase crystal structures, we have used pharmacophore and docking studies to elucidate a new binding pocket for isoprenoid analogues. The unique conformations between the first two isoprene units of 2Z-GGPP, but not GGPP, allows 2Z-GGPP to exploit this new binding pocket. The discovered conformation allows the molecule to adopt a reactive conformation while placing hydrophobic groups within the predominately hydrophobic binding pocket. This computational finding is supported by NMR studies on (13)C-labeled 2Z-farnesol, which confirm that the computationally predicted conformation is also favored in solution. These discoveries suggest that ligand conformational flexibility may be an important design consideration for the development of both inhibitors and alternative substrates of FTase. PMID- 16045301 TI - Surface-integral QSPR models: local energy properties. AB - Surface-integral models based on AM1 semiempirical molecular orbital calculations are presented for the free energies of solvation in water, n-octanol, and chloroform and for the enthalpy of solvation in water. A parametrized function of four local properties calculated at the isodensity surface (the molecular electrostatic potential, local ionization energy, electron affinity, and polarizability) is integrated over the triangulated surface area to obtain the target quantity. The resulting models give results only slightly less accurate than those reported for parametrized generalized Born/polar surface area models despite relying only on gas-phase calculations. The water and octanol free-energy models were validated by calculating the water-octanol partition coefficient for a test set of organic compounds with moderate success. The models lead to a local solvation energy, which can be projected onto the molecular isodensity surface and provides insight into "hot" areas for solvation in water or the other solvents. PMID- 16045302 TI - Retrospective docking study of PDE4B ligands and an analysis of the behavior of selected scoring functions. AB - Scoring forms a major obstacle to the success of any docking study. In general, fast scoring functions perform poorly when used to determine the relative affinity of ligands for their receptors. In this study, the objective was not to rank compounds with confidence but simply to identify a scoring method which could provide a 4-fold hit enrichment in a screening sample over random selection. To this end, LigandFit, a fast shape matching docking algorithm, was used to dock a variety of known inhibitors of type 4 phosphodiesterase (PDE4B) into its binding site determined crystallographically for a series of pyrazolopyridine inhibitors. The success of identifying good poses with this technique was explored through RMSD comparisons with 19 known inhibitors for which crystallographic structures were available. The effectiveness of five scoring functions (PMF, JAIN, PLP2, LigScore2, and DockScore) was then evaluated through consideration of the success in enriching the top ranked fractions of nine artificial databases, constructed by seeding 1980 inactive ligands (pIC50 < 5) with 20 randomly selected inhibitors (pIC50 > 6.5). PMF and JAIN showed high average enrichment factors (greater than 4 times) in the top 5-10% of the ranked databases. Rank-based consensus scoring was then investigated, and the rational combination of 3 scoring functions resulted in more robust scoring schemes with (cScore)-DPmJ (consensus score of DockScore, PMF, and JAIN) and (cScore)-PPmJ (PLP2, PMF, and JAIN) yielding particularly good results. These cScores are believed to be of greater general application. Finally, the analysis of the behavior of the scoring functions across different chemotypes uncovered the inherent bias of the docking and scoring toward compounds in the same structural family as that employed for the crystal structure, suggesting the need to use multiple versions of the binding site for more successful virtual screening strategies. PMID- 16045303 TI - Molecular modeling studies focused on 5-HT7 versus 5-HT1A selectivity. Discovery of novel phenylpyrrole derivatives with high affinity for 5-HT7 receptors. AB - The present study discusses the well-known 5-HT7/5-HT1A selectivity issue through a new series of phenylpyrrole derivatives. The first hits emerged from a virtual screening performed on a chemolibrary. Further study led to an optimization of a preliminary 5-HT7 pharmacophore model. The importance of each pharmacophoric feature is confirmed, but these characteristics have to be coupled to geometric constraints in order to achieve a 5-HT7 selectivity. Indeed, 5-HT1A affinity probably arises from extended conformations, whereas a bent one appears to be best suited for 5-HT7 selectivity. PMID- 16045304 TI - Ligand-based virtual screening and in silico design of new antimalarial compounds using nonstochastic and stochastic total and atom-type quadratic maps. AB - Malaria has been one of the most significant public health problems for centuries. It affects many tropical and subtropical regions of the world. The increasing resistance of Plasmodium spp. to existing therapies has heightened alarms about malaria in the international health community. Nowadays, there is a pressing need for identifying and developing new drug-based antimalarial therapies. In an effort to overcome this problem, the main purpose of this study is to develop simple linear discriminant-based quantitative structure-activity relationship (QSAR) models for the classification and prediction of antimalarial activity using some of the TOMOCOMD-CARDD (TOpological MOlecular COMputer Design Computer Aided "Rational" Drug Design) fingerprints, so as to enable computational screening from virtual combinatorial datasets. In this sense, a database of 1562 organic chemicals having great structural variability, 597 of them antimalarial agents and 965 compounds having other clinical uses, was analyzed and presented as a helpful tool, not only for theoretical chemists but also for other researchers in this area. This series of compounds was processed by a k-means cluster analysis in order to design training and predicting sets. Afterward, two linear classification functions were derived in order to discriminate between antimalarial and nonantimalarial compounds. The models (including nonstochastic and stochastic indices) correctly classify more than 93% of the compound set, in both training and external prediction datasets. They showed high Matthews' correlation coefficients, 0.889 and 0.866 for the training set and 0.855 and 0.857 for the test one. The models' predictivity was also assessed and validated by the random removal of 10% of the compounds to form a new test set, for which predictions were made using the models. The overall means of the correct classification for this process (leave group 10% full-out cross validation) using the equations with nonstochastic and stochastic atom-based quadratic fingerprints were 93.93% and 92.77%, respectively. The quadratic maps based TOMOCOMD-CARDD approach implemented in this work was successfully compared with four of the most useful models for antimalarials selection reported to date. The developed models were then used in a simulation of a virtual search for Ras FTase (FTase = farnesyltransferase) inhibitors with antimalarial activity; 70% and 100% of the 10 inhibitors used in this virtual search were correctly classified, showing the ability of the models to identify new lead antimalarials. Finally, these two QSAR models were used in the identification of previously unknown antimalarials. In this sense, three synthetic intermediaries of quinolinic compounds were evaluated as active/inactive ones using the developed models. The synthesis and biological evaluation of these chemicals against two malaria strains, using chloroquine as a reference, was performed. An accuracy of 100% with the theoretical predictions was observed. Compound 3 showed antimalarial activity, being the first report of an arylaminomethylenemalonate having such behavior. This result opens a door to a virtual study considering a higher variability of the structural core already evaluated, as well as of other chemicals not included in this study. We conclude that the approach described here seems to be a promising QSAR tool for the molecular discovery of novel classes of antimalarial drugs, which may meet the dual challenges posed by drug resistant parasites and the rapid progression of malaria illnesses. PMID- 16045305 TI - A virtual screening approach for thymidine monophosphate kinase inhibitors as antitubercular agents based on docking and pharmacophore models. AB - Docking and pharmacophore screening tools were used to examine the binding of ligands in the active site of thymidine monophosphate kinase of Mycobacterium tuberculosis. Docking analysis of deoxythymidine monophosphate (dTMP) analogues suggests the role of hydrogen bonding and other weak interactions in enzyme selectivity. Water-mediated hydrogen-bond networks and a halogen-bond interaction seem to stabilize the molecular recognition. A pharmacophore model was developed using 20 dTMP analogues. The pharmacophoric features were complementary to the active site residues involved in the ligand recognition. On the basis of these studies, a composite screening model that combines the features from both the docking analysis and the pharmacophore model was developed. The composite model was validated by screening a database spiked with 47 known inhibitors. The model picked up 42 of these, giving an enrichment factor of 17. The validated model was used to successfully screen an in-house database of about 500,000 compounds. Subsequent screening with other filters gave 186 hit molecules. PMID- 16045306 TI - Interpreting computational neural network quantitative structure-activity relationship models: a detailed interpretation of the weights and biases. AB - In this work, we present a methodology to interpret the weights and biases of a computational neural network (CNN) quantitative structure-activity relationship model. The methodology allows one to understand how an input descriptor is correlated to the predicted output by the network. The method consists of two parts. First, the nonlinear transform for a given neuron is linearized. This allows us to determine how a given neuron affects the downstream output. Next, a ranking scheme for neurons in a layer is developed. This allows us to develop interpretations of a CNN model similar in manner to the partial least squares (PLS) interpretation method for linear models described by Stanton. The method is tested on three datasets covering both physical and biological properties. The results of this interpretation method correspond well to PLS interpretations for linear models using the same descriptors as the CNN models, and they are consistent with the generally accepted physical interpretations for these properties. PMID- 16045307 TI - Lead hopping using SVM and 3D pharmacophore fingerprints. AB - The combination of 3D pharmacophore fingerprints and the support vector machine classification algorithm has been used to generate robust models that are able to classify compounds as active or inactive in a number of G-protein-coupled receptor assays. The models have been tested against progressively more challenging validation sets where steps are taken to ensure that compounds in the validation set are chemically and structurally distinct from the training set. In the most challenging example, we simulate a lead-hopping experiment by excluding an entire class of compounds (defined by a core substructure) from the training set. The left-out active compounds comprised approximately 40% of the actives. The model trained on the remaining compounds is able to recall 75% of the actives from the "new" lead series while correctly classifying >99% of the 5000 inactives included in the validation set. PMID- 16045308 TI - Consensus scoring criteria for improving enrichment in virtual screening. AB - MOTIVATION: Virtual screening of molecular compound libraries is a potentially powerful and inexpensive method for the discovery of novel lead compounds for drug development. The major weakness of virtual screening-the inability to consistently identify true positives (leads)-is likely due to our incomplete understanding of the chemistry involved in ligand binding and the subsequently imprecise scoring algorithms. It has been demonstrated that combining multiple scoring functions (consensus scoring) improves the enrichment of true positives. Previous efforts at consensus scoring have largely focused on empirical results, but they have yet to provide a theoretical analysis that gives insight into real features of combinations and data fusion for virtual screening. RESULTS: We demonstrate that combining multiple scoring functions improves the enrichment of true positives only if (a) each of the individual scoring functions has relatively high performance and (b) the individual scoring functions are distinctive. Notably, these two prediction variables are previously established criteria for the performance of data fusion approaches using either rank or score combinations. This work, thus, establishes a potential theoretical basis for the probable success of data fusion approaches to improve yields in in silico screening experiments. Furthermore, it is similarly established that the second criterion (b) can, in at least some cases, be functionally defined as the area between the rank versus score plots generated by the two (or more) algorithms. Because rank-score plots are independent of the performance of the individual scoring function, this establishes a second theoretically defined approach to determining the likely success of combining data from different predictive algorithms. This approach is, thus, useful in practical settings in the virtual screening process when the performance of at least two individual scoring functions (such as in criterion a) can be estimated as having a high likelihood of having high performance, even if no training sets are available. We provide initial validation of this theoretical approach using data from five scoring systems with two evolutionary docking algorithms on four targets, thymidine kinase, human dihydrofolate reductase, and estrogen receptors of antagonists and agonists. Our procedure is computationally efficient, able to adapt to different situations, and scalable to a large number of compounds as well as to a greater number of combinations. Results of the experiment show a fairly significant improvement (vs single algorithms) in several measures of scoring quality, specifically "goodness-of-hit" scores, false positive rates, and "enrichment". This approach (available online at http://gemdock.life. nctu.edu.tw/dock/download.php) has practical utility for cases where the basic tools are known or believed to be generally applicable, but where specific training sets are absent. PMID- 16045309 TI - Predicting the phosphorylation sites using hidden Markov models and machine learning methods. AB - Accurately predicting phosphorylation sites in proteins is an important issue in postgenomics, for which how to efficiently extract the most predictive features from amino acid sequences for modeling is still challenging. Although both the distributed encoding method and the bio-basis function method work well, they still have some limits in use. The distributed encoding method is unable to code the biological content in sequences efficiently, whereas the bio-basis function method is a nonparametric method, which is often computationally expensive. As hidden Markov models (HMMs) can be used to generate one model for one cluster of aligned protein sequences, the aim in this study is to use HMMs to extract features from amino acid sequences, where sequence clusters are determined using available biological knowledge. In this novel method, HMMs are first constructed using functional sequences only. Both functional and nonfunctional training sequences are then inputted into the trained HMMs to generate functional and nonfunctional feature vectors. From this, a machine learning algorithm is used to construct a classifier based on these feature vectors. It is found in this work that (1) this method provides much better prediction accuracy than the use of HMMs only for prediction, and (2) the support vector machines (SVMs) algorithm outperforms decision trees and neural network algorithms when they are constructed on the features extracted using the trained HMMs. PMID- 16045310 TI - Inversion of the rectifying effect in diblock molecular diodes by protonation. AB - A new molecular diode based on biphenyl-co-bispyrimidine was synthesized and showed a pronounced rectifying effect. Further studies indicate that protonation on the nitrogen atoms of the diode molecule by strong acids can reversibly alter the rectifying direction. This phenomenon can be envisioned as a starting point for single molecule detection devices. PMID- 16045311 TI - Reaction of hydroxyfullerene with metal salts: a route to remediation and immobilization. AB - Hydroxylated fullerene reacts rapidly and irreversibly (across a wide pH range) with Fe(NO3)3, Al(NO3)3, CaCl2, CoCl2, CuCl2, KMnO4, Ag(NO3), and ZnCl2 under ambient aqueous conditions to produce insoluble metal-hydroxyfullerene cross linked polymers (M-fullerenol). Materials have been characterized by SEM, TEM, AFM, XPS, and UV-visible spectroscopy. Molecular mechanics calculations on the model systems, [Fe(C60O2)2] and [Fe(C60O2)3], show that both tetrahedral and octahedral coordination are possible. The rate of precipitation reaction is proportional to the concentration of both reagents. The interaction of hydroxyfullerenes with metals is an important issue with regard to waste treatment, fullerene exposure in the environment, and fullerene-based pharmaceutical agents. PMID- 16045312 TI - Synthesis and characterization of beta-haloalkenyl-lambda3-bromanes: stereoselective Markovnikov addition of difluoro(aryl)-lambda3-bromane to terminal acetylenes. AB - Reported here for the first time are the synthesis, isolation, and characterization of hypervalent beta-haloalkenyl-lambda3-bromanes. Exposure of terminal alkynes to p-trifluoromethylphenyl(difluoro)-lambda3-bromane activated by BF3-i-Pr2O resulted in fluoro-lambda3-bromanation of the triple bonds in a Markovnikov fashion, yielding (E)-beta-fluoroalkenyl-lambda3-bromanes stereoselectively in good yields. 5-Chloro-1-pentynes undergo domino lambda3 bromanation-chlorine shift-fluorination or lambda3-bromanation-chlorine shift alkyl shift-fluorination reaction, depending on the substituents and afford (E) beta-chloroalkenyl-lambda3-bromanes stereoselectively in high yields. The beta chloroalkenyl-lambda3-bromanes contain three kinds of halogen atoms, F, Cl, and Br, in the molecule. PMID- 16045313 TI - Stepwise growth of a single polymer chain. AB - By monitoring the modulation of an ionic current passing through a nanoreactor formed from a protein pore, the step-by-step growth of an individual polymer chain was monitored. The observation of polymer growth at the single-molecule level will be useful for studying the kinetics of chain growth or the movement of polymers under confinement. It might also be used to synthesize "molecular fishing lines" in situ, for applications in stochastic sensing. PMID- 16045314 TI - Ion sensing coupled to resonance energy transfer: a highly selective and sensitive ratiometric fluorescent chemosensor for Ag(I) by a modular approach. AB - We report a novel dimeric boradiazaindacene dye which can be converted in one step to an efficient resonance energy transfer (RET) dyad. In addition, if this modification is done with appropriate ligands, RET can be coupled to ion sensing. The utility of this approach is demonstrated in a highly selective, emission ratiometric chemosensor for Ag(I). PMID- 16045315 TI - Laminar flow-based electrochemical microreactor for efficient regeneration of nicotinamide cofactors for biocatalysis. AB - One of the long-standing challenges in biocatalysis is the search for methods to continuously regenerate essential cofactors such as NADH that would enable a wide range of enzymes to be used in the more environmentally friendly synthesis of chiral fine chemicals including pharmaceuticals, cosmetics, and food additives. This communication reports a microreactor-based cofactor regeneration method that exploits the microfluidic phenomenon of laminar flow: a reactant stream and a buffer stream are introduced in a microchannel and continue to flow side by side without turbulent mixing between two electrodes that cover opposing channel walls. Adjustment of the flow rate ratio of the two streams in laminar flow enables focusing of the reactant stream close to the cathode, thereby reversing a normally unfavorable reaction equilibrium essential for cofactor regeneration. The absence of a bulk phase in these microreactors prevents the undesired reverse reaction to take place, which has prevented the use of electrochemical cofactor regeneration in macroscale processes. Here, we demonstrate the regeneration of NADH with conversion efficiencies as high as 31%. We also show the subsequent in situ conversion of an achiral substrate, pyruvate, into a chiral product, l lactate, within this microreactor. PMID- 16045316 TI - Real-time membrane fusion of giant polymer vesicles. AB - Membrane fusion is very important for the formation of many complex organs in metazoans throughout evolution, such as muscles, bones, and placentae. Lipid vesicles (liposomes) are frequently used as model membranes to study the fusion process. This work demonstrates for the first time the real-time membrane fusion of giant polymer vesicles by directly displaying a series of high-resolution and real-time transformation images of individual vesicles. The fusion process includes the sequential steps of membrane contact, forming the center wall, symmetric expansion of fusion pore and complete fusion, undergoing the intermediates of "8" shape with a protruding rim at the contact site, peanut (pear) shape, and oblate sphere. The vesicle swells during fusion, and the fusing vesicle only deforms in the neck domain around the fusion pore in the lateral direction, which verifies the importance of the lateral tension on the fusion pore at the vesicle deformation level. The successful fusion of the synthetic and protein-free polymer vesicles reported here also supports that vesicle proximity combined with membrane perturbation suffices to induce membrane fusion, and that the protein is not necessary for the fusion process. PMID- 16045317 TI - A new synthetic approach to phenol derivatives: use of ring-closing olefin metathesis. AB - Phenol derivatives, which are one of the most important classes of aromatic compounds in organic chemistry, were synthesized by ruthenium-catalyzed ring closing olefin metathesis (RCM) of 1,4,7-trien-3-ones with versatile substitution patterns. The RCM reaction for producing phenol derivatives was also successful with 1,5,7-trien-3-one as another precursor. Most of the phenols prepared here could not be obtained easily by conventional methods. PMID- 16045318 TI - Substrate binding modulates the reduction potential of DNA photolyase. AB - The reduction potential of the (FADH-/FADH*) couple in DNA photolyase was measured, and the value was found to be significantly higher than the values estimated in the literature. In the absence of substrate, the enzyme has a reduction potential of 16 +/- 6 mV vs NHE. In the presence of excess substrate the reduction potential increases to 81 +/- 8 mV vs NHE. The increase in reduction potential has physiological relevance since it gives the catalytic state greater resistance to oxidation. This is the first measurement of a reduction potential for this class of DNA-repair enzymes and the larger family of blue-light photoreceptors. PMID- 16045319 TI - Kinetic resolution of axially chiral 2,2'-dihydroxy-1,1'-biaryls by palladium catalyzed alcoholysis. AB - Palladium-diamine complexes catalyzed kinetic resolution of axially chiral 2,2' dihydroxy-1,1'-biaryls by alcoholysis of vinyl ethers. The reaction proceeded with high selectivity for various kinds of biaryls. This process is applicable to not only binaphthols but also biphenols, which have been considered to be difficult for the enantioselective synthesis by known catalytic methods. PMID- 16045320 TI - The Ni-mediated cyclocarbonylation of allyl halides and alkynes made catalytic. Evidence supporting the involvement of pseudoradical Ni(i) species in the mechanism. AB - We report here on a highly efficient catalytic method to synthesize intermolecularly the cyclopentane skeleton from starting products as simple as allyl halides, alkynes, and carbon monoxide under very mild reaction conditions by means of a substoichiometric amount of iron, acetone, and a catalytic amount of Ni(II) iodide. PMID- 16045321 TI - Palladium pincer complex catalyzed substitution of vinyl cyclopropanes, vinyl aziridines, and allyl acetates with tetrahydroxydiboron. An efficient route to functionalized allylboronic acids and potassium trifluoro(allyl)borates. AB - Palladium-catalyzed boronation of vinyl cyclopropane, vinyl aziridine, and allyl acetate substrates could be accomplished using tetrahydroxydiboron reagent in the presence of SeCSe pincer complex catalyst 1a. These reactions result in allyl boronic acids, which were converted to synthetically useful trifluoro(allyl)borates or allyl boronates. The catalytic transformations proceed under mild and neutral conditions, and therefore many functionalities Br, COOEt, ArSO2(NH), OAc, and SiRMe2 are tolerated. The selectivity of the presented processes is very high, affording the linear products incorporating a trans double bond. PMID- 16045322 TI - Ultrastable Au nanocatalyst supported on surface-modified TiO2 nanocrystals. AB - The surfaces of TiO2 nanocrystals were modified with amorphous aluminum-oxide layers using a surface sol-gel process to control the interaction between supports and metal particles. Ultrastable Au nanocatalysts were prepared by the deposition of Au nanoparticles on the surface-modified TiO2 nanocrystals using a deposition-precipitation (DP) method. The TEM analysis showed that the Au nanoparticles on the surface-modified nanocrystal supports were highly stable with a sinter-resistant capability during high-temperature calcination. The HRTEM analysis revealed that the surface of the TiO2 nanocrystals was covered by an amorphous aluminum-oxide layer and the Au nanoparticles were primarily anchored to this amorphous layer. This amorphous aluminum-oxide layer played an extremely important role in the stabilization of the supported Au nanoparticles without affecting catalytic activities. The surface modification of nanocrystal supports highlights new opportunities in tailoring the stability and activity of supported nanocatalyst systems. PMID- 16045323 TI - Catalytic enantioselective Negishi reactions of racemic secondary benzylic halides. AB - This report describes the first enantioselective cross-couplings of racemic secondary benzylic halides, specifically, nickel-catalyzed Negishi reactions of bromides and chlorides. The catalyst components are commercially available and air-stable, and the reaction is not highly oxygen- or moisture-sensitive (it can be set up in the air). The method has been applied to the catalytic enantioselective synthesis of intermediates employed by others in the generation of bioactive compounds (e.g., trikentrin A and an androgen receptor agonist). PMID- 16045324 TI - Nanoparticles on the basis of highly functionalized dextrans. AB - New nanoparticles based on well-defined dextran esters were prepared by a dialysis process. Dextran was converted into a propionate with a degree of substitution of 1.70 and, subsequently, acylated under homogeneous reaction conditions with a pyroglutamic acid imidazolide, which is prepared in situ by conversion of pyroglutamic acid with N,N-carbonyldiimidazole. The synthesis path allows perfect control of the amphiphilicity and solubility. The highly functionalized polysaccharide derivative avoids the collapse of the nanostructure due to the prevention of hydrogen bond formation. The major fraction of the dextran propionate pyroglutamate nanoparticles investigated by SEM and AFM exhibits narrow size distribution with 370 nm as mean diameter and uniform spherical shape. The SEM image verifies that polymeric nanoparticles in the suspensions did not undergo any morphological changes within 3 weeks. PMID- 16045325 TI - Efficient complexation of N-acetyl amino acid carboxylates in water by an artificial receptor: unexpected cooperativity in the binding of glutamate but not aspartate. AB - A new tris-cation 1 binds N-acetyl amino acid carboxylates in water even at millimolar salt concentrations with Kass approximately 103 M-1 due to a clustering of electrostatic interactions. Binding is efficient enough to allow a naked-eyed detection using an indicator displacement assay. Furthermore, receptor 1 shows an unexpected 2:1 complex formation with strong positive cooperativity with glutamate but not aspartate. PMID- 16045326 TI - Modular monodentate phosphoramidite ligands for rhodium-catalyzed enantioselective hydrogenation. AB - A new class of monodentate phosphoramidite ligands (DpenPhos) has been developed on the basis of the modular concept for Rh(I)-catalyzed asymmetric hydrogenations of a variety of olefin derivatives, affording the corresponding optically active compounds in excellent yields and enantioselectivities. The ligands have the advantages of facile preparation, tunable structure, and broad scope of substrates in their Rh(I) complex-catalyzed asymmetric hydrogenations. PMID- 16045327 TI - Identification of partially disordered peptide intermediates through residue specific NMR diffusion measurements. AB - It is technically challenging to detect low-population partially disordered species that are in equilibrium with the folded and unfolded states. Residue specific translational diffusion experiments measured by pulsed field gradient NMR have been used to detect the presence, and define the conformation, of such equilibrium intermediates. The experiment is demonstrated for equilibrium solutions of related triple helical peptides that model a small region of type I collagen with and without a mutation known to cause osteogenesis imperfecta. The data show that residue-specific diffusion coefficients of an interconverting trimer to monomer system can allow discrimination between a simple two-state model and more complex multistate models involving partially disordered intermediates. PMID- 16045328 TI - Allosterically activated Diels-Alder catalysis by a ribozyme. AB - We describe the allosteric control of Diels-Alder reactions by a small organic effector, theophylline. This is achieved by converting a Diels-Alder ribozyme into an allosterically regulated system. In contrast to other published systems, we have a bond-forming reaction with two small-molecule substrates and multiple turnover. This system could be very attractive for the development of assays for a variety of analytes and can be regarded as a prototype of fully synthetic signaling cascades. PMID- 16045329 TI - Proximity-induced catalysis by the protein kinase ERK2. AB - Five hundred protein kinases phosphorylate 10 000 proteins in human cells. Frequently, more than one site in a protein is phosphorylated, and often by more than one protein kinase. The mechanistic basis underlying the overlapping specificity of the phospho-proteome is not well understood. We are interested in understanding why ERK2, a proline-directed protein kinase, phosphorylates only a fraction of the (S/T-P) sites found in the surface loops of proteins, at an appreciable rate. To address this fundamental question, we utilized a well established protein substrate EtsDelta138, which comprises a globular ERK2 recognition domain (pnt domain) and an unstructured peptide-like N-terminal tail. This tail contains T38, the sole ERK2 phosphorylation site. We mutated the TP motif, which is recognized by the active site and found that mutagenesis of the T 38/P-39 motif to TD, TR, TA, TG, and TV has no effect on the stability of the ternary complex but does decrease kcat. We also investigated the effect of perturbing the binding between ERK2 and the pnt domain, which occurs outside the active site, to find that mutation of the pnt domain (F120A) leads to a 10-fold decrease in binding but the kcat remains the same. The data support a mechanism of proximity-mediated catalysis, where the docking of the pnt domain, outside the active site, increases the effective concentration of the TP motif near the active site. The data are consistent with the notion that the interaction between ERK2 and the pnt domain provides uniform binding energy and stabilizes each enzyme intermediate and transition state to an equal extent. While other steps on the reaction pathway contribute towards the specificity of the ERK2 reaction, a docking interaction provides the initial basis for substrate recognition. Those residues within the docked complex, which have the ability to access the active site with an appropriate geometry, can be phosphorylated at an efficient rate if followed by a proline or small hydrophobic amino acid. PMID- 16045330 TI - Nanobelt self-assembly from an organic n-type semiconductor: propoxyethyl-PTCDI. AB - Nanobelt structures have been fabricated for an n-type semiconductor molecule, N,N'-di(propoxyethyl)perylene-3,4,9,10-tetracarboxylic diimide (PTCDI). The short alkyloxy side chain not only affords effective pi-pi stacking in polar solvents for self-assembling but also provides sufficient solubility in nonpolar solvents for solution processing. As revealed by both AFM and electron microscopies, the nanobelts have an approximately rectangular cross section, with a typical thickness of about 100 nm and a width in the range of 300-500 nm. The length of the nanobelts ranges from 10 to a few tens of micrometers. The highly organized molecular packing (uniaxial crystalline phase) has been deduced from the measurement of electron diffraction and polarized microscopy imaging. The detected optical axis is consistent with the one-dimensional stacking of the molecules. PMID- 16045331 TI - Interfacial polymerization within a simplified microfluidic device: capturing capsules. AB - A simple approach to a microfluidic device is described. The device is composed of flexible tubing and a needle inserted orthogonal to the long axis of the tubing. This design is well suited to creating oil-water interfaces allowing the formation of laminar flows and monodisperse emulsions. The system is characterized by mapping the phases observed as a function of organic phase flow and Reynolds number. In addition, the device allows interfacial polymerization reactions to capture low coefficient of variation capsules. The shell structure and surface are examined by scanning electron microscopy. PMID- 16045332 TI - 1,2-Halogen migration in haloallenyl ketones: regiodivergent synthesis of halofurans. AB - Selective 1,2-iodine, bromine, and chlorine migration in haloallenyl ketones in the presence of Au catalyst has been demonstrated. It was found that, depending on the nature of the Au catalyst used, either selective bromine migration or hydrogen shift occurs, leading to the formation of 3- or 2-bromofurans, respectively. Halirenium intermediate was proposed for the unusual 1,2-halogen migration. This cascade transformation allows for mild and efficient synthesis of various types of 3-halofurans. PMID- 16045333 TI - Synthesis and structure of fused alpha-oligothiophenes with up to seven rings. AB - To combine the stability of alpha-oligothiophenes with the planarity of acenes, fully fused oligothienoacenes were synthesized and their properties compared to the nonfused alpha-oligothiophenes. By employing removable solubilizing groups, our synthetic methodology made it possible to efficiently prepare and purify oligothienoacenes with up to seven fused rings. The key steps involved the halogen dance reaction and Pd-catalyzed coupling of Bu3SnSSnBu3 to introduce sulfur linkages. This approach eliminates alpha-beta anion equilibration, a significant improvement over the traditional method of introducing sulfur linkages via Li-Br exchange. X-ray diffraction data indicate that pentathienoacene and heptathienoacene adopt pi-stacked packing motifs in contrast to the herringbone packing of nonfused oligothiophenes. On the basis of the linear dependence of the longest lambdamax on the reciprocal number of double bonds of thienoacenes with three to seven rings, the band gap of polythienoacene is extrapolated to be 2.21 eV. PMID- 16045334 TI - Design of an organocatalyst for the enantioselective Diels-Alder reaction with alpha-acyloxyacroleins. AB - We have realized the first enantioselective organocatalytic Diels-Alder reaction between alpha-substituted acroleins, such as alpha-acyloxyacroleins, and not only cyclic but also acyclic dienes. alpha-Acyloxyacroleins are useful as synthetic equivalents of alpha-haloacroleins. The present catalyst could be prepared in situ from pentafluorobenzenesulfonic acid (2.5-3.0 equiv) and chiral triamine (1 equiv) derived from H-l-Phe-l-Leu-N(CH2CH2)2. The enantioselective Diels-Alder reaction of 5-(benzyloxymethyl)cyclopentadiene, cyclopentadiene, cyclohexadiene, 2,3-dimethylbutadiene, and isoprene with alpha-(p-methoxybenzoyloxy)acrolein catalyzed by the above chiral ammonium salt (2.5-20 mol %) at -20-22 degrees C gave the corresponding adducts with 83, 83, 91, 92, and 88% ee, respectively. PMID- 16045335 TI - Ni(II) Tol-BINAP-catalyzed enantioselective orthoester alkylations of N acylthiazolidinethiones. AB - The development of a Ni(II) (S)-Tol-BINAP-catalyzed enantioselective alkylation of N-acylthiazolidinethiones with trimethyl orthoformate is described. The reaction is general with respect to the thiazolidinethione, providing the desired adducts in good to excellent yield and with high levels of enantioselectivity (11 examples, 51-92% yield, 90-99% ee). The products are readily converted into a variety of synthetically useful derivatives in a straightforward manner. PMID- 16045336 TI - A synthetic breakthrough into an unanticipated stability regime: readily isolable complexes in which C16-C28 polyynediyl chains span two platinum atoms. AB - The oxidative cross-coupling of trans-(p-tol)(p-tol3P)2Pt(CC)2H (PtC4H) and excess H(CC)2SiEt3 (O2, cat. CuCl, TMEDA, acetone; Hay conditions) gives PtC8Si (29%), PtC12Si (30%), and PtC16Si (1%; Si = SiEt3). The less stable PtC6H is generated in situ from PtC6Si and n-Bu4N+F-; following the addition of ClSiMe3 (F scavenger) and excess H(CC)2SiEt3, the Hay conditions afford PtC10Si (59%) and PtC14Si (7%). Analogous sequences can be conducted with PtC8Si and PtC10Si. When PtC6Si, PtC8Si, and PtC10Si are similarly reacted in the absence of H(CC)2SiEt3, homocouplings to PtC12Pt (88%), PtC16Pt (70%), and PtC20Pt (72%) occur. However, analogous reactions of PtC12Si and PtC14Si fail, presumably due to the rapid decomposition of PtC12H and PtC14H. Bronsted acidity trends suggest that (CC)n moieties should become better leaving groups with increasing chain length. Thus, when PtC12Si and PtC14Si are subjected to the Hay conditions alone, PtC24Pt (36%) and PtC28Pt (51%) are isolated, with substantial recovery of starting material from the former reaction. Presumably adventitious water and/or other nucleophiles effect desilylation (as also seen in the cross-couplings), after which homocoupling is rapid due to the simultaneous availability of an oxidizing agent. PtC24Pt and PtC28Pt are thermally stable to >/=140 degrees C, whereas all other known dodecaynes and tetradecaynes rapidly decompose at room temperature. There is every reason to believe that this series can be further extended. UV-visible spectra show progressively red-shifted and more intense bands with epsilon > 400 000 M-1 cm-1. PMID- 16045337 TI - DNA interstrand cross-link formation initiated by reaction between singlet oxygen and a modified nucleotide. AB - DNA is the target of many anti-cancer therapies. These agents damage the biopolymer by oxidation or by alkylation. Interstrand DNA cross-links are believed to be the source of cytotoxicity of anti-tumor agents, such as mitomycin C, which alkylate the biopolymer. In contrast, deoxyguanosine oxidation is the result of reaction between DNA and singlet oxygen, which is the damaging species produced in photodynamic therapy. We have shown that, upon oxidation by singlet oxygen, an analogue of thymidine (2) rearranges to a methide, which forms DNA-DNA interstrand cross-links. This novel process suggests that 2 may be a useful adjuvant in photodynamic therapy. PMID- 16045338 TI - High-valent nonheme iron. Two distinct iron(IV) species derived from a common iron(II) precursor. AB - The reaction of [Fe(II)(beta-BPMCN)(OTf)2] (1, BPMCN = N,N'-bis(2-pyridylmethyl) N,N'-dimethyl-trans-1,2-diaminocyclohexane) with tBuOOH at low-temperature yields alkylperoxoiron(III) intermediates 2 in CH2Cl2 and 2-NCMe in CH3CN. At -45 degrees C and above, 2-NCMe converts to a pale green species 3 (lambda(max) = 753 nm, epsilon = 280 M(-1) cm(-1)) in 90% yield, identified as [Fe(IV)(O)(BPMCN)(NCCH3)]2+ by comparison to other nonheme [Fe(IV)(O)(L)]2+ complexes. Below -55 degrees C in CH2Cl2, 2 decays instead to form deep turquoise 4 (lambda(max) = 656, 845 nm; epsilon = 4000, 3600 M(-1) cm(-1)), formulated to be an unprecedented alkylperoxoiron(IV) complex [Fe(IV)(BPMCN)(OH)(OOtBu)]2+ on the basis of Mossbauer, EXAFS, resonance Raman, NMR, and mass spectral evidence. The reactivity of 1 with tBuOOH in the two solvents reveals an unexpectedly rich iron(IV) chemistry that can be supported by the BPMCN ligand. PMID- 16045339 TI - Engineering InAs(x)P(1-x)/InP/ZnSe III-V alloyed core/shell quantum dots for the near-infrared. AB - Quantum dots with a core/shell/shell structure consisting of an alloyed core of InAs(x)P(1-x), an intermediate shell of InP, and an outer shell of ZnSe were developed. The InAs(x)P(1-x) alloyed core has a graded internal composition with increasing arsenic content from the center to the edge of the dots. This compositional gradient results from two apparent effects: (1) the faster reaction kinetics of the phosphorus precursor compared to the arsenic precursor, and (2) a post-growth arsenic-phosphorus exchange reaction that increases the arsenic content. The cores have a zinc blend structure for all compositions and show tunable emission in the near-infrared (NIR) region. A first shell of InP leads to a red-shift and an increase in quantum yield. The final shell of ZnSe serves to stabilize the dots for applications in aqueous environments, including NIR biomedical fluorescence imaging. These NIR-emitting core/shell/shell InAs(x)P(1 x)/InP/ZnSe were successfully used in a sentinel lymph node mapping experiment. PMID- 16045340 TI - Sensitivity of ammonia interaction with single-walled carbon nanotube bundles to the presence of defect sites and functionalities. AB - Ammonia adsorption on single-walled carbon nanotubes (SWNTs) was studied by means of infrared spectroscopy at both cryogenic (approximately 94 K) and room (approximately 300 K) temperatures. At 94 K, vacuum-annealed SWNTs showed no detectable ammonia uptake. However, the ammonia adsorption was found to be sensitive to the functionalities and defects on the nanotube surfaces. NH3 adsorption was detected on HNO3-treated nanotubes, characterized by significant functionalities and defects, prior to vacuum annealing. NH3 desorbed from those nanotubes above 140 K, indicating a weak adsorbate-nanotube interaction (approximately 30 kJ/mol). Exposure of annealed samples to ambient air, which possibly regenerated functionalities and defects on nanotube surfaces, restored partially the ammonia uptake capacity. No ammonia adsorption on SWNTs was observed by infrared spectroscopy at room temperature with up to 80 Torr dosing pressure. This work suggests the influence of functionalities and/or defect densities on the sensitivity of SWNT chemical gas sensors. Our theoretical studies on NH3 adsorption on pristine and defective tubes, as well as oxidized tubes, corroborate these findings. PMID- 16045341 TI - Sterically directed functionalization of aromatic C-H bonds: selective borylation ortho to cyano groups in arenes and heterocycles. AB - Ir-catalyzed borylations of 4-substituted benzonitriles are described. In contrast to electrophilic aromatic substitutions and directed ortho metalations, C-H activation/borylation enables functionalization at the 2-position, adjacent to the cyano group, when the 4-subsitutent is larger than cyano. When an excess of borane reagent is used, diborylation can be achieved with a single regioisomer being formed in certain cases. Extension of sterically directed borylation to cyano-substituted, five- and six-membered ring heterocycles is also reported. PMID- 16045342 TI - Crystal structure prediction of aminols: advantages of a supramolecular synthon approach with experimental structures. AB - The supramolecular synthon approach to crystal structure prediction (CSP) takes into account the complexities inherent in crystallization. The synthon is a kinetically favored unit, and through analysis of commonly occurring synthons in a group of related compounds, kinetic factors are implicitly invoked. The working assumption is that while the experimental structure need not be at the global minimum, it will appear somewhere in a list of computationally generated structures so that it can be suitably identified and ranked upward using synthon information. These ideas are illustrated with a set of aminophenols, or aminols. In the first stage, a training database is created of the 10 isomeric methylaminophenols. The crystal structures of these compounds were determined. The prototypes 2-, 3-, and 4-aminophenols were also included in the training database. Small and large synthons in these 13 crystal structures were then identified. Small synthons are of high topological but low geometrical value and are used in negative screens to eliminate computationally derived structures that are chemically unreasonable. Large synthons are more restrictive geometrically and are used in positive screens ranking upward predicted structures that contain these more well-defined patterns. In the second stage, these screens are applied to CSP of nine new aminols carried out in 14 space groups. In each space group, up to 10 lowest energy structures were analyzed with respect to their synthon content. The results are encouraging, and the predictions were classified as good, unclear, or bad. Two predictions were verified with actual crystal structure determinations. PMID- 16045343 TI - Amphiphilic guest sorption of K2[Cr3O(OOCC2H5)6(H2O)3]2[alpha-SiW12O40] ionic crystal. AB - An ionic crystal K2[Cr3O(OOCC2H5)6(H2O)3]2[alpha-SiW12O40] x 3H2O (1a) is synthesized by the complexation of a Keggin-type polyoxometalate of [alpha SiW12O40]4- with K+ and a macrocation of [Cr3O(OOCC2H5)6(H2O)3]+. Compound 1a possesses both hydrophilic and hydrophobic channels in the crystal lattice. The 3 mol mol(-1) of the water of crystallization in 1a resides in the hydrophilic channel. The water of crystallization is removed by the evacuation at 303 K to form the guest-free phase 1b with small changes in the lattice lengths (+/-0.2 A). The water sorption profile is reproduced by the single rate constant. Therefore, the water sorbed probably resides in the hydrophilic channel. Compound 1b sorbs various kinds of polar organic molecules, and the amounts of < or = C3 alcohols are comparable to or larger than that of water, while chlorocarbons with no hydrogen-bonding ability and nonpolar molecules are excluded. Thus, 1b showed the amphiphilic sorption property. The states of the polar organic molecules sorbed in 1b have been quantitatively investigated using ethanol as a probe molecule. The IR, NMR, and single-crystal X-ray diffraction studies combined with the sorption kinetics reveal that ethanol molecules are mainly sorbed into the hydrophilic channel at P/P0 < or = 0.5, while the sorption into the hydrophobic channel is dominant at P/P0 > or = 0.6. Thus, it is demonstrated that ethanol molecules enter both hydrophilic and hydrophobic channels of 1b. PMID- 16045344 TI - Facile Diels-Alder reactions with pyridines promoted by tungsten. AB - The isoquinuclidine (2-azabicyclo[2.2.2]octane) core is found in numerous molecules of biological and medicinal importance, including the widely investigated Iboga alkaloids and their related bisindole Cantharanthus alkaloids (Sundberg, R. J.; Smith, S. Q. Alkaloids (San Diego, CA, United States) 2002, 59, 281-386). A diverse range of synthetic methods for the stereoselective construction of this architecture is required for the efficient development of related pharmaceuticals. Here, we report a fundamentally new methodology that constructs the isoquinuclidine core directly from pyridines, using a pi-basic tungsten complex to disrupt the aromatic stabilization of these otherwise inert heterocycles. By this approach, common pyridines are found to undergo stereoselective Diels-Alder reactions with electron-deficient alkenes under mild reaction conditions, thus providing access to a broad range of functionalized isoquinuclidines. Further, by using the common terpene alpha-pinene, a single enantiomer of the tungsten fragment can be isolated and used to provide access to enantio-enriched isoquinuclidines from pyridines. PMID- 16045345 TI - Computational and ESR studies of electron attachment to decafluorocyclopentane, octafluorocyclobutane, and hexafluorocyclopropane: electron affinities of the molecules and the structures of their stable negative ions as determined from 13C and 19F hyperfine coupling constants. AB - High-resolution ESR spectra of the ground-state negative ions of hexafluorocyclopropane (c-C3F6*-), octafluorocyclobutane (c-C4F8*-), and decafluorocyclopentane (c-C5F10*-) are reported and their isotropic 19F hyperfine coupling constants (hfcc) of 198.6 +/- 0.4 G, 147.6 +/- 0.4 G, and 117.9 +/- 0.4 G, respectively, are in inverse ratio to the total number of fluorine atoms per anion. Together with the small value of 5.2 +/- 0.4 G determined for the isotropic 13C hfcc of c-C4F8*-, these results indicate that in each case the singly occupied molecular orbital (SOMO) is delocalized over the equivalent fluorines and possesses a nodal plane through the carbon atoms of a time-averaged D(nh) structure. A series of quantum chemical computations were carried out to further characterize these anions and their neutral counterparts. Both the B3LYP density functional and second-order Moller-Plesset perturbation theory (MP2) indicate that c-C3F6*- adopts a D(3h) geometry and a (2)A2'' ground electronic state, that c-C4F8*- adopts a D(4h) geometry and a (2)A2u ground electronic state, and that c-C5F10*- adopts a C(s) structure and a (2)A' electronic state. Moreover, the 19F hyperfine coupling constants computed with the MP2 method and a high quality triple-zeta basis set are within 1% of the experimental values. Also, the values computed for the 13C hfcc of c-C4F8*- are consistent with the experimental value of 5.2 G. Therefore, in keeping with the ESR results, these negative ions derived from first-row elements can be characterized as pi* species. In addition, the hypervalency of these perfluorocycloalkane radical anions has been clarified. PMID- 16045346 TI - Putative DNA quadruplex formation within the human c-kit oncogene. AB - The DNA sequence, d(AGGGAGGGCGCTGGGAGGAGGG), occurs within the promoter region of the c-kit oncogene. We show here, using a combination of NMR, circular dichroism, and melting temperature measurements, that this sequence forms a four-stranded quadruplex structure under physiological conditions. Variations in the sequences that intervene between the guanine tracts have been examined, and surprisingly, none of these modified sequences forms a quadruplex arrangement under these conditions. This suggests that the occurrence of quadruplex-forming sequences within the human and other genomes is less than was hitherto expected. The c-kit quadruplex may be a new target for therapeutic intervention in cancers where there is elevated expression of the c-kit gene. PMID- 16045347 TI - Abiotic metallofoldamers as electrochemically responsive molecules. AB - Described are the design, synthesis, and study of nonbiological molecules based on salophen and salen ligands that fold into single-stranded helices in the presence of either Ni(II) or Cu(II). X-ray diffraction studies show that the materials fold into helical structures in the solid state, and a series of NMR studies provide strong evidence that the folded structures are conserved in solution. Metal coordination is required for folding, as NMR and X-ray show that the free ligands do not adopt helical structures. Two of the racemic metallofoldamers spontaneously resolve during crystallization from CHCl3/acetonitrile, and CD spectroscopy and optical rotation show that the resolved, crystalline materials racemize quickly when dissolved at 5 degrees C. This shows that the secondary structures can reorganize easily and can, therefore, provide the basis for responsive materials. By comparison, an analogue from enantiomerically pure (R,R)-(-)-trans-cyclohexanediamine showed a strong CD signal and a large specific rotation. Electrochemical experiments show that a structural reorganization occurs upon metal-centered reduction of a Cu(II) containing foldamer. When the reduction is carried out in the presence of coordinating ligands, it is proposed that apical binding of those ligands gives square pyramidal complexes. Semiempirical (AM1) calculations support that the helical structure would be disrupted by the reduction to Cu(I) with concomitant reorganization to a square pyramidal complex. PMID- 16045348 TI - Photoinduced charge transfer processes along triarylamine redox cascades. AB - In this paper, we describe the synthesis and photophysical properties of a series of acridine-triarylamine redox cascades. These cascades were designed in order to promote photoinduced hole transfer from an acridine fluorophore into an adjacent triarylamine. The excited dipolar state then injects a hole into the triarylamine redox cascade. Subsequently, the hole migrates along the redox gradient which was tuned by the substituents attached to the triarylamine redox centers. The rate of hole migration was determined by fluorescence lifetime measurements and is in the ns regime and depends strongly on the solvent polarity. The photophysical processes were also investigated by femtosecond broadband pump-probe spectroscopy. Our studies reveal different dynamic processes in the cascades depending on the solvent polarity, e.g., direct charge separation after photoexcitation vs a two step hole transfer mechanism. PMID- 16045349 TI - Optical spectra of delocalized dinitroaromatic radical anions revisited. AB - The optical spectra of nine dinitroaromatic radical anions (1,2- and 1,4 dinitrobenzene, 1,5- and 2,6-dinitro naphthalene, 4,4'-dinitrobiphenyl, 2,7 dinitro-9,9-dimethylfluorene, 2,6-dinitroanthracene, and 2,7- and 1,8 dinitrobiphenylene) in dimethylformamide are reported and analyzed. All have delocalized charge distribution, as demonstrated by the vibrational fine structure that is observed in their optical spectra: All show lowest energy absorption bands that correspond to an alpha-homo (highest occupied molecular orbital) to alpha-lumo (lowest unoccupied) transition, as shown by Koopmans-based UB3LYP calculation of the orbital separation of the neutral at the geometry of the radical anion. These single-point calculations are shown to be significantly more accurate for five of these compounds than the much more complex and expensive TD-DFT method. The two-state model should not be used to estimate the electronic coupling in delocalized intervalence compounds such as these. Neighboring orbital estimation of the electronic couplings show that using the two-state model greatly underestimates electronic coupling here. PMID- 16045350 TI - Ligand-stabilized aromatic three-membered gold rings and their sandwichlike complexes. AB - Electronic structure calculations (DFT) suggest that ligand-stabilized three membered gold(I) rings constituting the core structure in a series of cyclo Au3L(n)H(3-n) (L = CH3, NH2, OH and Cl; n = 1, 2, 3) molecules exhibit aromaticity, which is primarily due to 6s and 5d cyclic electron delocalization over the triangular Au3 framework (s- and d-orbital aromaticity). The aromaticity of the novel triangular gold(I) isocycles was verified by a number of established criteria of aromaticity. In particular, the nucleus-independent chemical shift, NICS(0), the upfield changes in the chemical shifts for Li+, Ag+, and Tl+ cations over the Au3 ring plane, and their interaction with electrophiles (e.g., H+, Li+, Ag+, and Tl+) are indicative for the aromaticity of the three-membered gold(I) rings. Interestingly, unlike the respective substituted derivatives of cyclopropenium cation and the bora-cyclopropene carbacyclic analogues, the aromatic Au3 rings, although exhibit comparable diatropicity, react with electrophiles in a different way affording 1:1 and 2:1 sandwichlike complexes. The bonding in the three-membered gold(I) rings is characterized by a common ring shaped electron density, more commonly seen in aromatic organic molecules and in "all-metal" aromatics, such as the cyclo-[Hg3]4- tetraanion. Moreover, the cation pi interactions in the 1:1 and 1:2 sandwichlike complexes formed upon reacting the Au3 rings with electrophiles, depending on the nature of the cation, are predicted to be predominantly electrostatic (Li+, Tl+) or covalent (H+, Ag+). The 1:2 complexes constitute a new class of sandwichlike complexes, which are expected to have novel properties and applications. PMID- 16045351 TI - Electrochemical templating of metal nanoparticles and nanowires on single-walled carbon nanotube networks. AB - The use of single-walled carbon nanotube (SWNT) networks as templates for the electrodeposition of metal (Ag and Pt) nanostructures is described. Pristine SWNTs, grown on insulating SiO2 surfaces using catalyzed chemical vapor deposition, served as the working electrode. In the simplest case, electrical contact was made by depositing a gold strip on the SWNT substrate (device 1). Deposition of Ag and Pt over extensive periods (30 s) resulted in a high density of particles on the SWNTs, with almost contiguous nanowire formation from the Au/SWNT boundary moving to isolated nanoparticles at further distances from the contact. For direct electrochemical studies of Ag and Pt nucleation, the assembly was coated in a resist layer and a small window opened up to expose only the electrically connected SWNTs to solution (device 2). In this case, the electrochemical signature in voltammetric and amperometric studies of metal deposition was due solely to processes at the SWNTs. Coupled with high-resolution microscopy measurements (atomic force microscopy and field emission scanning electron microscopy), this approach provided detail on the nucleation and growth mechanisms of Ag and Pt on SWNTs under electrochemical control. In particular, Ag growth was found to be rapid and progressive with an increasing nanoparticle density with time, whereas Pt deposition was characterized by lower nucleation densities and slower growth rates with a tendency for larger particles to be produced over long times. PMID- 16045352 TI - On the nature of the transition state in catechol O-methyltransferase. A complementary study based on molecular dynamics and potential energy surface explorations. AB - The way in which enzymes influence the rate of chemical processes is still a question of debate. The protein promotes the catalysis of biochemical processes by lowering the free energy barrier in comparison with the reference uncatalyzed reaction in solution. In this article we are reporting static and dynamic aspects of the enzyme catalysis in a bimolecular reaction, namely a methyl transfer from S-adenosylmethionine to the hydroxylate oxygen of a substituted catechol catalyzed by catechol O-methyltransferase. From QM/MM optimizations, we will first analyze the participation of the environment on the transition vector. The study of molecular dynamics trajectories will allow us to estimate the transmission coefficient from a previously localized transition state as the maximum in the potential of mean force profile. The analysis of the reactive and nonreactive trajectories in the enzyme environment and in solution will also allow studying the geometrical and electronic changes, with special attention to the chemical system movements and the coupling with the environment. The main result, coming from both analyses, is the approximation of the magnesium cation to the nucleophilic and the hydroxyl group of the catecholate as a result of a general movement of the protein, stabilizing in this way the transition state. Consequently, the free energy barrier of the enzyme reaction is dramatically decreased with respect to the reaction in solution. PMID- 16045353 TI - A joint experimental and theoretical study of cation-pi interactions: multiple decker sandwich complexes of ferrocene with alkali metal ions (Li+, Na+, K+, Rb+, Cs+). AB - A systematic study of cation-pi interactions between alkali metal ions and the cyclopentadienyl ring of ferrocene is presented. The alkali metal (Li+, Na+, K+, Rb+, Cs+) salts of the ditopic mono(pyrazol-1-yl)borate ligand [1,1' fc(BMe2pz)2]2- crystallize from dimethoxyethane as multiple-decker sandwich complexes with the M+ ions bound to the pi faces of the ferrocene cyclopentadienyl rings in an eta5 manner (fc = (C5H4)2Fe; pz = pyrazolyl). X-ray crystallography of the lithium complex reveals discrete trimetallic entities with each lithium ion being coordinated by only one cyclopentadienyl ring. The sodium salt forms polyanionic zigzag chains where each Na+ ion bridges the cyclopentadienyl rings of two ferrocene moieties. Linear columns [-CpR-Fe-CpR-M+ CpR-Fe-CpR-M+-](infinity) (R = [-BMe2pz]-) are established by the K+, Rb+, and Cs+ derivatives in the solid state. According to DFT calculations, the binding enthalpies of M+-eta5(ferrocene) model complexes are about 20% higher as compared to the corresponding M+-eta6(benzene) aggregates when M+ = Li+ or Na+. For K+ and Rb+, the degree of cation-pi interaction with both aromatics is about the same. The binding sequence along the M+-eta5(ferrocene) series follows a classical electrostatic trend with the smaller ions being more tightly bound. PMID- 16045354 TI - Mycosamine orientation of amphotericin B controlling interaction with ergosterol: sterol-dependent activity of conformation-restricted derivatives with an amino carbonyl bridge. AB - Amphotericin B (AmB 1) is known to assemble together and form an ion channel across biomembranes. The antibiotic consists of mycosamine and macrolactone moieties, whose relative geometry is speculated to be determinant for the drug's channel activity and sterol selectivity. To better understand the relationship between the amino-sugar orientation and drug's activity, we prepared conformation restricted derivatives 2-4, in which the amino and carboxyl groups were bridged together with various lengths of alkyl chains. K+ influx assays across the lipid bilayer membrane revealed that ergosterol selectivity was markedly different among derivatives; short-bridged derivative 2 almost lost the selectivity, while 3 showed higher ergosterol preference than AmB itself. Monte Carlo conformational analysis of 2-4 based on NOE-derived distances indicated that the amino-sugar moiety of 2 comes close to C41 because of the short bridge, whereas those of 3 and 4 are pointing outward. The mutual orientation of the amino-sugar moiety and macrolide ring is so rigid in derivatives 2 and 3 that these conformations should be unchanged upon complex formation in lipid membranes. These results strongly suggest that the large difference in sterol preference between derivatives 2 and 3 is ascribed to the different orientation of amino-sugar moieties. These findings allowed us to propose a simple model accounting for AmB-sterol interactions, in which hydrogen bonding between 2'-OH of AmB and 3beta-OH of ergosterol plays an important role. PMID- 16045355 TI - Generation of superparamagnetic liposomes revealed as highly efficient MRI contrast agents for in vivo imaging. AB - Maghemite (gamma-Fe2O3) nanocrystals stable at neutral pH and in isotonic aqueous media were synthesized and encapsulated within large unilamellar vesicles of egg phosphatidylcholine (EPC) and distearoyl-SN-glycero-3-phosphoethanolamine-N [methoxy(poly(ethylene glycol))-2000] (DSPE-PEG(2000), 5 mol %), formed by film hydration coupled with sequential extrusion. The nonentrapped particles were removed by flash gel exclusion chromatography. The magnetic-fluid-loaded liposomes (MFLs) were homogeneous in size (195 +/- 33 hydrodynamic diameters from quasi-elastic light scattering). Iron loading was varied from 35 up to 167 Fe(III)/lipid mol %. Physical and superparamagnetic characteristics of the iron oxide particles were preserved after liposome encapsulation as shown by cryogenic transmission electron microscopy and magnetization curve recording. In biological media, MFLs were highly stable and avoided ferrofluid flocculation while being nontoxic toward the J774 macrophage cell line. Moreover, steric stabilization ensured by PEG-surface-grafting significantly reduced liposome association with the macrophages. The ratios of the transversal (r2) and longitudinal (r1) magnetic resonance (MR) relaxivities of water protons in MFL dispersions (6 < r2/r1 < 18) ranked them among the best T2 contrast agents, the higher iron loading the better the T2 contrast enhancement. Magnetophoresis demonstrated the possible guidance of MFLs by applying a magnetic field gradient. Mouse MR imaging assessed MFLs efficiency as contrast agents in vivo: MR angiography performed 24 h after intravenous injection of the contrast agent provided the first direct evidence of the stealthiness of PEG-ylated magnetic-fluid-loaded liposomes. PMID- 16045356 TI - Water-assisted oxo mechanism for heme metabolism. AB - A mechanism of heme metabolism by heme oxygenase (HO) is discussed from B3LYP density functional theory calculations. The concerted OH group attack to the alpha-carbon by the iron-hydroperoxo species is investigated using a model with full protoporphyrin IX to confirm our previous conclusion that this species does not have sufficient oxidizing power for heme oxidation (J. Am. Chem. Soc. 2004, 126, 3672). Calculated activation energies and structures of the intermediates and transition state for this process remain unchanged from those for a small model with porphine in the previous study, which shows that the inclusion of the side chain of the porphyrin ring is not essential in describing the OH group transfer. The activation barrier for a direct oxo attack to the alpha-carbon by an iron-oxo model is calculated to be 49.8 kcal/mol, the barrier height of which looks very high for the enzymatic reaction under physiological conditions. This large activation energy is due to a highly bent porphyrin structure in the transition state. However, a bridging water molecule plays an important role in reducing the porphyrin distortion in the transition state, resulting in a remarkable decrease of the activation barrier to 13.9 kcal/mol. A whole-enzyme model with about 4000 atoms is constructed to elucidate functions of the protein environment in this enzymatic reaction using QM/MM calculations. The key water molecule is fixed in the protein environment to ensure the low-barrier and regioselective heme oxidation. A water-assisted oxo mechanism of heme oxidation by heme oxygenase is proposed from these calculational results. PMID- 16045357 TI - Noncovalent modification of chymotrypsin surface using an amphiphilic polymer scaffold: implications in modulating protein function. AB - We report here on a new amphiphilic homopolymer that binds noncovalently to proteins. This polymer not only binds to the target protein chymotrypsin with submicromolar affinity but also stabilizes the native structure of the protein. Since the polymer-protein binding process is based on electrostatic interaction, the bound protein can be released from the polymer surface and reactivated either by increasing the ionic strength or by adding complementary cationic surfactants. The electrostatic binding of polymer to the protein results in a marked change in the substrate specificity of chymotrypsin. PMID- 16045358 TI - Can the pi-facial selectivity of solvation be predicted by atomistic simulation? AB - This work is concerned with the rationalization and prediction of solvent and temperature effects in nucleophilic addition to alpha-chiral carbonyl compounds leading to facial diastereoselectivity. We study, using molecular dynamics simulations, the facial solvation of (R)-2-phenyl-propionaldehyde in n-pentane and n-octane at a number of temperatures and compare it with experimental selectivity data for the nBuLi addition leading to syn- and anti-(2R)-2-phenyl-3 heptanol, which give nonlinear Eyring plots with the presence of inversion temperatures. We have found from simulations that the facial solvation changes with temperature and alkane. Moreover, by introducing a suitable molecular chirality index we have been able to predict break temperatures (T(CI)) for the two solvents within less than 20 degrees of the inversion temperatures experimentally observed in the diastereoselective nBuLi addition. We believe this could lead to a viable approach for predicting inversion temperatures and other subtle solvent effects in a number of stereoselective reactions. PMID- 16045359 TI - Microfabrication of three-dimensional bioelectronic architectures. AB - The functionality and structural diversity of biological macromolecules has motivated efforts to exploit proteins and DNA as templates for synthesis of electronic architectures. Although such materials offer promise for numerous applications in the fabrication of cellular interfaces, biosensors, and nanoelectronics, identification of techniques for positioning and ordering bioelectronic components into useful patterns capable of sophisticated function has presented a major challenge. Here, we describe the fabrication of electronic materials using biomolecular scaffolds that can be constructed with precisely defined topographies. In this approach, a tightly focused pulsed laser beam capable of promoting protein photo-cross-linking in specified femtoliter volume elements is scanned within a protein solution, creating biomolecular matrices that either remain in integral contact with a support surface or extend as free standing structures through solution, tethered at their ends. Once fabricated, specific protein scaffolds can be selectively metallized via targeted deposition and growth of metal nanoparticles, yielding high-conductivity bioelectronic materials. This aqueous fabrication strategy opens new opportunities for creating electronic materials in chemically sensitive environments and may offer a general approach for creating microscopically defined inorganic landscapes. PMID- 16045360 TI - Cyano- and isocyanotris(trifluoromethyl)borates: syntheses, spectroscopic properties, and solid state structures of K[(CF3)3BCN] and K[(CF3)3BNC]. AB - A two step synthesis to the isocyanotris(trifluoromethyl)borate anion, [(CF3)3BNC]-, and its isomerization to the cyanotris(trifluoromethyl)borate anion, [(CF3)3BCN]-, at temperatures above 150 degrees C are presented. In the first step (CF3)3BNCH was obtained by reacting (CF3)3BCO with hydrogen cyanide followed by deprotonation of the HCN adduct with Li[N(SiMe3)2] in toluene. The thermal behavior of K[(CF3)3BNC] and K[(CF3)3BCN] were investigated by differential scanning calorimetry (DSC), and K[BF4] was identified as a major solid decomposition product. The enthalpy of the isocyanide-cyanide rearrangement, deltaH(iso) = -35 +/- 4 kJ mol(-1), was obtained from DSC measurements, and the activation energy, E(a) = 180 +/- 20 kJ mol(-1), from kinetic measurements. The isomerization was modeled as an intramolecular reaction employing DFT calculations at the B3LYP/6-311+G(d) level of theory yielding a reaction enthalpy of deltaH(iso) = -36.1 kJ mol(-1) and an activation energy of E(a) = 155.7 kJ mol(-1). The solid-state structures of K[(CF3)3BNC] and K[(CF3)3BCN] were determined by single-crystal X-ray diffraction. Both salts are isostructural and crystallize in the orthorhombic space group Pnma (no. 62). In the crystals the borate anions possess C(s) symmetry, while for the energetic minimum C3 symmetry is predicted by DFT calculations. The borate anions have been characterized by IR and Raman spectroscopy as well as by NMR spectroscopy. The assignment of the IR and Raman bands is supported by their calculated wavenumbers and intensities. The spectroscopic and structural properties of both borate anions are compared to the properties of the isoelectronic borane carbonyl (CF3)3BCO and the [B(CF3)4]- anion as well as to those of other related species. PMID- 16045361 TI - Mechanistic investigations of the ethylene tetramerisation reaction. AB - The unprecedented selective tetramerisation of ethylene to 1-octene was recently reported. In the present study various mechanistic aspects of this novel transformation were investigated. The unusually high 1-octene selectivity in chromium-catalyzed ethylene tetramerisation reactions is caused by the unique extended metallacyclic mechanism in operation. Both 1-octene and higher 1-alkenes are formed by further ethylene insertion into a metallacycloheptane intermediate, whereas 1-hexene is formed by elimination from this species as in other reported trimerisation reactions. This is supported by deuterium labeling studies, analysis of the molar distribution of 1-alkene products, and identification of secondary co-oligomerization reaction products. In addition, the formation of two C6 cyclic products, methylenecyclopentane and methylcyclopentane, is discussed, and a bimetallic disproportionation mechanism to account for the available data is proposed. PMID- 16045362 TI - X-ray diffraction and DOSY NMR characterization of self-assembled supramolecular metallocyclic species in solution. AB - Wide-angle X-ray scattering and diffusion NMR techniques have been used to obtain structural information on three self-assembled metallacyclic supramolecular complexes in solution: a rectangle, a triangle, and a three-diminsional cage. The low-angle region of the measured diffraction patterns and hydrodynamic radii calculations, determined from DOSY NMR experiments, suggest that the supramolecular assemblies retain their shape when dissolved in nitromethane. The experimental structure functions for the large-angle region have been analyzed, and the intramolecular contributions of the platinum-platinum interactions are discussed. These scattering measurements provide evidence that the supramolecular assemblies are not as rigid in solution as they are in the single crystal. Finally, by analysis of the radial distribution functions of the solutions, direct structural information (e.g., platinum-platinum intramolecular distances and coordination number) about the supramolecular assemblies has been obtained. PMID- 16045363 TI - Substrate discrimination by cholapod anion receptors: geometric effects and the "affinity-selectivity principle". AB - Cholapod anion receptors can achieve high affinities while maintaining compatibility with nonpolar media. Previously they have been shown to transport anions across cell and vesicle membranes. In the present work, the scope of the architecture is expanded and structure-selectivity relationships are investigated. Eight new receptors have been synthesized, with up to six H-bond donor centers. Using Cram's extraction method, these compounds plus five known examples have been tested for binding to seven monovalent anions (tetraethylammonium salts, wet chloroform as solvent). Association constants in excess of 10(10) M(-1) have been measured for several pairings. Selectivities vary with receptor geometry, as expected. More remarkably, they also depend on receptor strength: more powerful receptors show a wider range of binding free energies, and therefore a greater spread of Ka(X-)/Ka(Y-). This "affinity selectivity" effect can be derived from empirical relationships for H-bond strengths, and could prove widely operative in supramolecular chemistry. PMID- 16045364 TI - A systematic investigation of the synthetic utility of glycopeptide glycosyltransferases. AB - Glycosyltransferases involved in the biosynthesis of bacterial secondary metabolites may be useful for the generation of sugar-modified analogues of bioactive natural products. Some glycosyltransferases have relaxed substrate specificity, and it has been assumed that promiscuity is a feature of the class. As part of a program to explore the synthetic utility of these enzymes, we have analyzed the substrate selectivity of glycosyltransferases that attach similar 2 deoxy-L-sugars to glycopeptide aglycons of the vancomycin-type, using purified enzymes and chemically synthesized TDP beta-2-deoxy-L-sugar analogues. We show that while some of these glycopeptide glycosyltransferases are promiscuous, others tolerate only minor modifications in the substrates they will handle. For example, the glycosyltransferases GtfC and GtfD, which transfer 4-epi-L vancosamine and L-vancosamine to C-2 of the glucose unit of vancomycin pseudoaglycon and chloroorienticin B, respectively, show moderately relaxed donor substrate specificities for the glycosylation of their natural aglycons. In contrast, GtfA, a transferase attaching 4-epi-L-vancosamine to a benzylic position, only utilizes donors that are closely related to its natural TDP sugar substrate. Our data also show that the spectrum of donors utilized by a given enzyme can depend on whether the natural acceptor or an analogue is used, and that GtfD is the most versatile enzyme for the synthesis of vancomycin analogues. PMID- 16045365 TI - Modeling of stimulated hydrogel volume changes in photonic crystal Pb2+ sensing materials. AB - We modeled the stimulated hydrogel volume transitions of a material which binds Pb2+ and is used as a photonic crystal chemical sensing material. This material consists of a polymerized crystalline colloidal array (PCCA) hydrogel which contains a crown ether molecular recognition group. The PCCA is a polyacrylamide hydrogel which embeds a crystalline colloidal array (CCA) of monodisperse polystyrene spheres of approximately 100 nm. The array spacing is set to diffract light in the visible spectral region. Changes in the hydrogel volume induced by Pb2+ binding alter the array spacing and shift the diffracted wavelength. This system allows us to sensitively follow the hydrogel swelling behavior which results from the immobilization of the Pb2+ by the crown ether chelating groups. Binding of the Pb2+ immobilizes its counterions. This results in a Donnan potential, which results in an osmotic pressure which swells the hydrogel. We continue here our development of a predictive model for hydrogel swelling based on Flory's theory of gel swelling. We are qualitatively able to model the PCCA swelling but cannot correctly model the large responsivity observed at the lowest Pb2+ concentrations which give rise to the experimentally observed low detection limits for Pb2+. These PCCA materials enable stimulated hydrogel volume transitions to be studied. PMID- 16045366 TI - Reactivity of monolayer-protected gold nanoclusters at dye-sensitized liquid/liquid interfaces. AB - Hexanethiolate monolayer-protected gold nanoclusters (MPCs) were used as redox quenchers at the polarizable water/1,2-dichloroethane (DCE) interface. Photocurrent responses originating from the heterogeneous quenching of photoexcited water soluble porphyrin complexes by MPCs dissolved in the DCE phase were observed. As MPCs can function as both electron acceptors and donors, the photocurrent results from the superposition of two simultaneous processes, which correspond to the oxidation and reduction of MPCs. The magnitude of the net photocurrent is essentially determined by the balance of the kinetics of these two processes, which can be controlled by tuning the Galvani potential difference between the two phases. We show that, within the available potential window, the apparent electron-transfer rate constants follow classical Butler-Volmer dependence on the applied potential difference. PMID- 16045367 TI - Total synthesis of ningalin D. AB - A concise (nine-step) and effective (19% overall yield) total synthesis of ningalin D (1a) is disclosed and is based on a key 1,2,4,5-tetrazine --> 1,2 diazine --> pyrrole Diels-Alder strategy to assemble a fully substituted pyrrole core central to its structure. Additional highlights of the synthesis include a double Dieckmann condensation to introduce the C and D aryl rings enlisting substituents judiciously placed on the dienophile and intrinsic to the widely used tetrazine 2, a highly effective Suzuki coupling of the resulting C and D phenol triflates for introduction of the sterically demanding F and G aryl rings, and an unusually effective formal oxidative decarboxylation reaction cascade initiated by a Curtius rearrangement to directly provide the biphenylene quinone methide found imbedded in the structure of ningalin D. The cytotoxic and multidrug resistance (MDR) reversal activity of ningalin D, its derivatives, and the key synthetic intermediates are detailed. PMID- 16045368 TI - Results of the healthy body healthy spirit trial. AB - Healthy Body Healthy Spirit was a multicomponent intervention to increase fruit and vegetable (F & V) consumption and physical activity (PA) delivered through Black churches. Sixteen churches were randomly assigned to 3 intervention conditions. At baseline, 1,056 individuals were recruited across the 16 churches, of which 906 (86%) were assessed at 1-year follow-up. Group 1 received standard educational materials, Group 2 received culturally targeted self-help nutrition and PA materials, and Group 3 received the same intervention as did Group 2 as well as 4 telephone counseling calls based on motivational interviewing (MI) delivered over the course of 1 year. At 1-year follow-up, Groups 2 and 3 showed significant changes in both F & V intake and PA. Changes were somewhat larger for F & V. For F & V, but not PA, there was a clear additive effect for the MI intervention. PMID- 16045370 TI - Medication adherence and diabetes control in urban African Americans with type 2 diabetes. AB - In 181 urban African Americans with Type 2 diabetes, medication adherence was assessed using a measure designed specifically for an urban, impoverished sociodemographic population. Hemoglobin A-sub(1c), blood pressure and cholesterol levels, medication-related beliefs, and depression were assessed. Seventy-four percent of the sample reported adherence to diabetes medication. Adherence, adjusted for age, was associated with lower hemoglobin A-sub(1c). The specific behaviors associated with poorer diabetes control were forgetting to take medications and running out of medications. Knowledge of blood glucose goals differed for adherers and nonadherers. Blood pressure and cholesterol medication adherence rates were not associated with actual levels of blood pressure or lipids, respectively. These data suggest that specific medication-taking behaviors are important to diabetes control and constitute logical targets for interventions. ((c) 2005 APA, all rights reserved). PMID- 16045371 TI - Using UV photography to reduce use of tanning booths: a test of cognitive mediation. AB - Two laboratory studies were conducted in which a new type of intervention was used to reduce ultraviolet radiation (UV) exposure from tanning booth use among college students (Time 1 Ns=70 and 134). The intervention uses UV photography to highlight the damage to facial skin caused by previous UV exposure. When the authors controlled for baseline measures of booth use, students in both studies who viewed their UV photographs reported less booth use at a follow-up session 3 4 weeks later than did students not shown a copy of their photograph. Also, in both studies, the decline in use was significantly mediated by a Tanning Cognition Index composed of variables suggested by the prototype-willingness (prototype) model of health risk: tanning attitudes, tanner prototypes, and willingness to engage in risky UV exposure. PMID- 16045372 TI - Testing a model of pain appraisal and coping in children with chronic abdominal pain. AB - This prospective study of children with recurrent abdominal pain (N=133; ages 8- 15 years) used path analysis to examine relations among dispositional pain beliefs and coping styles, cognitions and behavior related to a specific pain episode, and short- and long-term outcomes. Children believing they could not reduce or accept pain appraised their episode-specific coping ability as low and reported passive coping behavior. Dispositional passive coping had direct effects on both episode-specific passive coping and long-term symptoms and disability. Accommodative coping (acceptance and self-encouragement) was associated with reduced episode-specific distress, which itself predicted reduced depressive symptoms 3 months later. Results suggest that coping-skill interventions for children with chronic pain should target reductions in passive coping and consider the potential benefits of accommodative coping strategies. PMID- 16045373 TI - Central adiposity is associated with cardiovascular reactivity to stress in adolescents. AB - The current study examined the association between central adiposity, measured by waist circumference, and cardiovascular reactivity to stress among 106 White and 105 Black adolescents, approximately 50% of whom were girls. Participants engaged in 4 laboratory tasks while cardiovascular reactivity measures were taken. Independent of body mass index, race, and gender, participants with a greater waist circumference exhibited greater systolic blood pressure reactivity and diastolic blood pressure reactivity (boys only). Race did not affect the results. Results from the present study suggest that central adiposity is associated with blood pressure reactivity early in life, especially in adolescent boys. PMID- 16045374 TI - A stress and coping model of medication adherence and viral load in HIV-positive men and women on highly active antiretroviral therapy (HAART). AB - The authors tested a structural model that incorporated age, time since diagnosis, social support, coping, and negative mood as predictors of medication adherence and HIV viral load on 188 men and 134 women on highly active antiretroviral therapy (HAART). The authors used psychosocial latent factors formed from baseline measures to predict latent factors of adherence, as assessed by electronic monitoring and self-report, and viral load defined by indicators assessed over a 15-month period. Results from the model indicate that greater negative mood and lower social support are related to greater use of avoidance oriented coping strategies. Use of these coping strategies by patients on HAART is related to poorer medication adherence and, subsequently, higher viral load. This model advances researchers' understanding of the contribution of psychosocial variables in predicting treatment adherence and disease progression in HIV-positive men and women. PMID- 16045375 TI - Optimistic social comparisons of older adults low in primary control: a prospective analysis of hospitalization and mortality. AB - Social comparison can be used strategically to bolster the self-concept. Such use may constitute secondary striving for control, when primary striving through action is unattainable. On the basis of the life span theory of control, the authors hypothesized and found that social comparison judgments would predict physical health outcomes among older adults with low primary control perceptions in the health domain. Only among such respondents, after age, sex, activities of daily living, chronic conditions, and prior hospitalization were adjusted for, did more positive social comparison judgments predict significantly lower odds of hospitalization and death over the next 2--6 years as reported in provincial health records. In later life, optimistic social comparisons may contribute to better health by providing secondary control. PMID- 16045377 TI - Optimism and pessimism as predictors of change in health after death or onset of severe illness in family. AB - The authors prospectively examined changes in health after a major life event (death or onset of severe illness in family) among 5,007 employees (mean age=44.8 years) whose optimism and pessimism levels were assessed in 1997 and major life events in 2000. Health was indicated by sickness absence days during a period covering 36 months prior to the event and 18 months after the event. Increase in sick days after the event was smaller and returned to the prevent level more quickly among highly optimistic individuals than among their counterparts with low optimism. Parallel changes were not observed in relation to pessimism. These findings suggest that optimism may reduce the risk of health problems and may be related to a faster recovery after a major life event. PMID- 16045376 TI - Effects of a weight maintenance diet on bulimic symptoms in adolescent girls: an experimental test of the dietary restraint theory. AB - It is widely accepted that dieting increases the risk for bulimia nervosa, but there have been few experimental tests of this theory. The authors conducted a randomized experiment with adolescent girls (N=188) to examine the effects of a weight maintenance diet on bulimic symptoms. A manipulation check verified that the diet intervention resulted in weight maintenance and significantly reduced the risk for obesity onset and weight gain observed in assessment-only controls. As hypothesized, the diet intervention resulted in significantly greater decreases in bulimic symptoms and negative affect than observed in controls. These experimental findings, which converge with those from a weight loss diet experiment, appear antithetical to dietary restraint theory and suggest instead that dietary restriction curbs bulimic symptoms. PMID- 16045378 TI - Positive emotion and health: going beyond the negative. AB - This study examined the relationships between positive emotions and health. Two positive emotions were considered, hope and curiosity, in conjunction with 3 physician-diagnosed disease outcomes: hypertension, diabetes mellitus, and respiratory tract infections. Medical data were abstracted over a 2-year period from 1,041 patient records from a multispecialty medical practice, and emotions were assessed through a mailed questionnaire. Across 3 disease outcomes, higher levels of hope were associated with a decreased likelihood of having or developing a disease. Higher levels of curiosity were also associated with decreased likelihood of hypertension and diabetes mellitus. Results suggest that positive emotion may play a protective role in the development of disease. PMID- 16045379 TI - A dyadic investigation of exercise support between cardiac patients and their spouses. AB - The authors examined married partners' similarity in reported exercise behavior as a moderator of the association between social support for exercise provided and received by extending an actor-partner dyadic effects model. Participants were married cardiac rehabilitation patients and their spouses (N=99 couples). For couples similar in their reported exercise behavior, a significant association was found between both partners' independent reports of providing exercise support to and receiving exercise support from one another (n=49 couples). However, for couples differing in their reported exercise behavior (n=50 couples), no association was found between either partner's provision and receipt of support for exercise. Findings have the potential to inform practitioners of patients who may not be receiving adequate social support for their recommended exercise. Future interventions may consider implementing dyadic educational or motivational strategies with patients and their spouses. PMID- 16045380 TI - HIV-related stigma among market workers in China. AB - HIV-related stigma was examined among 209 employees and owners of stalls in 5 markets in an eastern coastal city in China. Of the participants 53% were women and 47% were men; 100% were Han. Ages ranged from 18 to 49 years (M=35, SD=8.1). Half of the participants believed that punishment was an appropriate response toward those living with HIV (50%). Over half (56%) were unwilling to be friends with infected individuals. The majority thought that those living with HIV should be isolated (73%). They agreed that persons living with HIV should not take care of other people's children (85%). Punishing beliefs toward persons living with HIV were related to being male, older, married, less educated, and unwilling to be tested for HIV. PMID- 16045381 TI - Changing behavior: theoretical development needs protocol adherence. PMID- 16045382 TI - Modulation of the effective salience of a stimulus by direct and associative activation of its representation. AB - In 2 experiments, rats received exposure to presentations of a footshock preceded by a given cue. In the PRf (partial reinforcement) condition, this cue also occurred in the absence of the shock; in the CRf (continuous reinforcement) condition, it did not. Subsequent testing in which a new stimulus was used to signal the shock (Experiment 1) showed that the shock was more effective as a reinforcer for the PRf than for the CRf group. In Experiment 2, the shock was used as a conditioned stimulus signaling food delivery, and it was found that conditioning occurred more readily in the PRf than in the CRf group. These results accord with the hypothesis that preexposure to the shock results in a decline in its effective salience but that experience of a cue that signals shock in the absence of the shock itself attenuates this effect and helps maintain stimulus salience. PMID- 16045383 TI - Spontaneous recovery of excitation but not inhibition. AB - Five magazine approach experiments with rats and 1 sign-tracking experiment with pigeons explored the possibility of an analogue to spontaneous recovery of excitatory conditioning from extinction: spontaneous recovery of inhibitory conditioning from training. Stimuli were 1st treated as conditioned inhibitors and then as conditioned exciters or as irrelevant to reinforcement. At issue was whether the passage of time after the 2nd treatment would allow partial restoration of the initial conditioned inhibition. The experiments differed in the design used to study recovery, the manner of reinforcing the inhibitor, the means of testing for recovery, the time interval allowed for recovery, and the species used. None of the experiments found evidence for recovery of the inhibitory learning with time, despite the concurrently measured presence of spontaneous recovery of excitatory conditioning after extinction. These experiments suggest that changes with time may be preferential to the learning that occurs during extinction. PMID- 16045384 TI - Imitative learning of stimulus-response and response-outcome associations in pigeons. AB - A novel automated procedure was used to study imitative learning in pigeons. In Experiments 1 and 2, observer pigeons witnessed a demonstrator pigeon successfully performing an instrumental discrimination in which different discriminative stimuli indicated which of 2 topographically distinct responses (R1 and R2) resulted in the delivery of seed. The observers were then presented with the discriminative stimuli and given access to the response panel. Observer pigeons' behavior during the discriminative stimuli was influenced by how the demonstrator had responded during these stimuli. In Experiment 3, observers witnessed demonstrator pigeons performing R1 for Outcome 1 and R2 for Outcome 2. Observers then received a procedure designed to devalue Outcome 1 relative to Outcome 2 and were subsequently less likely to perform R1 than R2. These results suggest that pigeons can learn both stimulus response and response-outcome associations by observation. PMID- 16045385 TI - Bidirectional associations in humans and rats. AB - In 1 experiment with humans and 3 experiments with rats, the authors sought evidence of temporal integration of independently acquired temporal relationships, including forward and backward temporal associations. The experiments were designed and analyzed in the framework of the temporal coding hypothesis (e.g., L. D. Matzel, F. P. Held, & R. R. Miller, 1988; H. I. Savastano & R. R. Miller, 1998) as a strategy toward illuminating the use of forward and backward temporal associations and assessing the directional nature of these temporal associations. The results obtained suggest that the temporal content of learning about paired events involves a single bidirectional association instead of 2 independent unidirectional, forward and backward, associations. PMID- 16045386 TI - Acquisition with partial and continuous reinforcement in rat magazine approach. AB - Partial reinforcement, compared with continuous reinforcement, is widely considered detrimental to Pavlovian conditioned responding. However, time accumulation models predict an invariance in acquisition when learning is assessed as a function of number of reinforcements, not trials, and intertrial interval is held constant. Three experiments examined this prediction in a rat magazine-approach procedure. All experiments showed superior responding with continuous reinforcement. Experiments 1 and 3 used common tests in between- and within-subject designs, respectively. Experiment 2 showed the same pattern in a discrimination. Earlier results are reanalyzed informally and in a meta-analysis. Contrary to previous summaries of the literature, evidence points to superior conditioned responding with continuous reinforcement in a number of procedures. Results are generally consistent with traditional associative models of learning. PMID- 16045387 TI - Motivational control of second-order conditioning. AB - Two experiments examined the motivational specificity of the associations that support 2nd-order conditioning. In the 1st phase of each experiment rats were exposed to 2 visual conditioned stimuli (CSs) paired with either a saline or food pellet unconditioned stimulus (US) prior to exposure to 2nd-order conditioning using 2 auditory CSs, 1 paired with each visual CS. Rats' motivational state was then shifted prior to a test such that if and only if specific motivational features of the 1st-order training US played a role in the 2nd-order associative structure would responding to the 2nd-order cues shift appropriately with the state change. Even when the US was irrelevant to the training motivational state, shifts in state revealed that it was encoded within the associative structure supporting 2nd-order responding. PMID- 16045388 TI - Perceptual learning enhances retrospective revaluation of conditioned flavor preferences in rats. AB - Forward and backward blocking of taste preference learning was compared in rats. In the forward condition, thirsty rats were exposed to a flavor (A) in sucrose solution (+) or in water (-), after which they were exposed to A in compound with another flavor (B) in sucrose solution (i.e., AB+). In the backward condition, these phases were reversed. Consumption of B alone was assessed when rats were food deprived. In the forward condition, rats given A+ consumed less B than rats given A-, providing evidence of forward blocking, whereas in the backward condition, rats given A+ drank more of B than those given A-. Subsequent experiments found that alternating but not blocked preexposure to A and B, when given prior to training, produced blocking of B whether A+ was given before or after AB+, suggesting that prior failures to observe backward blocking reflect failures of discrimination. PMID- 16045389 TI - Ordinal judgments and summation of nonvisible sets of food items by two chimpanzees and a rhesus macaque. AB - Two chimpanzees and a rhesus macaque rapidly learned the ordinal relations between 5 colors of containers (plastic eggs) when all containers of a given color contained a specific number of identical food items. All 3 animals also performed at high levels when comparing sets of containers with sets of visible food items. This indicates that the animals learned the approximate quantity of food items in containers of a given color. However, all animals failed in a summation task, in which a single container was compared with a set of 2 containers of a lesser individual quantity but a greater combined quantity. This difficulty was not overcome by sequential presentation of containers into opaque receptacles, but performance improved if the quantitative difference between sizes was very large. PMID- 16045390 TI - Associative change in connectionist networks: an addendum. AB - The results of a recent study have provided direct support for the suggestion that conditional learning in rats is best characterized by a 3-layer connectionist network (M. J. Allman, J. Ward-Robinson, & R. C. Honey, 2004). In the 2 experiments reported here, rats were used to investigate the nature of the changes that occur when a stimulus compound is presented, whose components activate hidden units associated with food and no food, and either food or no food is presented. The results of both experiments, while controlling for the possible contribution of associations between these hidden units (within-layer links), provide evidence that the distribution of associative change between units in the hidden layer that are activated by the stimulus compound and those in the output layer (between-layer links) are unequal. They also indicate that associative change is more marked on trials on which no food was presented than on trials on which food was presented. PMID- 16045391 TI - Sensory preconditioning in spatial learning using a touch screen task in pigeons. AB - The authors used a touch screen-based visual-search task to investigate spatial integration in pigeons. First, pigeons were presented with a consistent spatial relationship between compound visual landmarks (LMs) A-X and B-Y, separately. Next, pigeons learned to find a hidden goal on the monitor in the presence of LMs A and B. The goal bore a consistent spatial relationship to LM A, but not to LM B. On nonreinforced probe tests, the peak and distribution of responses to LM X suggest that pigeons computed a novel X-goal spatial relationship on the basis of X-A and A-goal spatial vectors. Responses to LM Y, however, revealed no evidence of spatial integration. These results replicate and extend those of A. P. Blaisdell and R. G. Cook (2005) using an open-field task. PMID- 16045392 TI - Applying bubbles to localize features that control pigeons' visual discrimination behavior. AB - The authors trained pigeons to discriminate images of human faces that displayed: (a) a happy or a neutral expression or (b) a man or a woman. After training the pigeons, the authors used a new procedure called Bubbles to pinpoint the features of the faces that were used to make these discriminations. Bubbles revealed that the features used to discriminate happy from neutral faces were different from those used to discriminate male from female faces. Furthermore, the features that pigeons used to make each of these discriminations overlapped those used by human observers in a companion study (F. Gosselin & P.G. Schyns, 2001). These results show that the Bubbles technique can be effectively applied to nonhuman animals to isolate the functional features of complex visual stimuli. PMID- 16045393 TI - Psychology at the intersection of work and family: recommendations for employers, working families, and policymakers. AB - Demographic data show that major changes have been occurring in the everyday lives of families over the last generation, with the majority of mothers of young children in the workforce and an increasing number of men and women assuming caregiving responsibilities for older relatives. Thus, the 2 primary identities of most adults, defined by their multiple family and work roles, need to be coordinated in ways that promote positive family outcomes, returns on investments for employers, and societal values. Despite changes in the workforce, the world of work is still largely organized for a family model that is increasingly rare- one with a stay-at-home caregiver. Recommendations based on psychological and other social science research are offered to align the needs of working families and employers. PMID- 16045394 TI - Positive psychology progress: empirical validation of interventions. AB - Positive psychology has flourished in the last 5 years. The authors review recent developments in the field, including books, meetings, courses, and conferences. They also discuss the newly created classification of character strengths and virtues, a positive complement to the various editions of the Diagnostic and Statistical Manual of Mental Disorders (e. g., American Psychiatric Association, 1994), and present some cross-cultural findings that suggest a surprising ubiquity of strengths and virtues. Finally, the authors focus on psychological interventions that increase individual happiness. In a 6-group, random assignment, placebo-controlled Internet study, the authors tested 5 purported happiness interventions and 1 plausible control exercise. They found that 3 of the interventions lastingly increased happiness and decreased depressive symptoms. Positive interventions can supplement traditional interventions that relieve suffering and may someday be the practical legacy of positive psychology. PMID- 16045395 TI - Jerome M. Sattler: Gold Medal Award for Life Achievement in the Application of Psychology. PMID- 16045396 TI - A. Eugene Shapiro: Gold Medal Award for Life Achievement in the Practice of Psychology. PMID- 16045397 TI - James J. Gallagher: Gold Medal Award for Life Achievement in Psychology in the Public Interest. PMID- 16045398 TI - Howard Leventhal: Gold Medal Award for Life Achievement in the Science of Psychology. PMID- 16045399 TI - Thomas E. Ludwig: Charles L. Brewer Award for Distinguished Teaching in Psychology. PMID- 16045412 TI - Linking decision-making research and cancer prevention and control: important themes. AB - This article describes 6 themes underlying the multiple presentations from the Basic and Applied Decision Making in Cancer Control meeting, held February 19-20, 2004. The following themes have important implications for research and practice linking basic decision-making research to cancer prevention and control: (a) Traditional decision-making theories fail to capture real-world decision making, (b) decision makers are often unable to predict future preferences, (c) preferences are often constructed on the spot and thus are influenced by situational cues, (d) decision makers often rely on feelings rather than beliefs when making a decision, (e) the perspective of the decision maker is critical in determining preferences, and (f) informed decision making may--or may not--yield the best decisions. PMID- 16045413 TI - Making better decisions: from measuring to constructing preferences. AB - The authors examine how a constructive preferences perspective might change the prevailing view of medical decision making by suggesting that the methods used to measure preferences for medical treatments can change the preferences that are reported. The authors focus on 2 possible techniques that they believe would result in better outcomes. The 1st is the wise selection of default options. Defaults may be best applied when strong clinical evidence suggests a treatment option to be correct for most people but preserving patient choice is appropriate. The 2nd is the use of environments that explicitly facilitate the optimal construction of preferences. This seems most appropriate when choice depends on a patient's ability to understand and represent probabilities and outcomes. For each technique, the authors describe the background and literature, provide a case study, and discuss applications. PMID- 16045414 TI - Decision making as coping. AB - This article reviews a model of emotional trade-off difficulty in decision making. The model argues that decision makers are motivated to cope with the negative emotion associated with decision-processing operations, notably emotion generated by explicit trade-offs between highly valued attributes. The article begins to explore implications of this model for patient decision making in the cancer control domain. For instance, the model points to emotional reactions to decisions as both a cost and a barrier in the move toward greater patient participation in health care decision making. PMID- 16045415 TI - Regret in cancer-related decisions. AB - Decision-related regret is a negative emotion associated with thinking about a past or future choice. The thinking component generally takes the form of a wish that things were otherwise and involves a comparison of what actually did or will take place with some better alternative--a "counterfactual thought." For predecisional (anticipated) regret, the thinking involves a mental simulation of the outcomes that might result from different choice options. Prior research has focused on regret associated with decision outcomes, addressing especially (a) the comparison outcome selected and (b) whether the outcome resulted from action or inaction. More recent research examines regret associated with the choice itself and with the preceding decision process. Interest here has focused on the justifiability of the choice made or the process used. In this article, the authors review current regret research and propose directions for extending it to cancer-related decisions. PMID- 16045416 TI - Basic and applied decision making in cancer control. AB - Decision making is fundamental to all aspects of cancer care--prevention, detection, treatment, survivorship, and end of life--yet researchers and clinicians have limited knowledge of the ways in which patients and their health care providers make critical health decisions. Recognizing how important it is to understand how patients and their providers make potentially life-altering decisions, the National Cancer Institute developed a decision making in cancer control initiative. The goal of this initiative is to enhance understanding of human decision-making processes so that individuals can make more informed and satisfying choices regarding their health. This article describes the multidisciplinary meeting that provided the scientific foundation for this initiative. PMID- 16045417 TI - Affect, risk, and decision making. AB - Risk is perceived and acted on in 2 fundamental ways. Risk as feelings refers to individuals' fast, instinctive, and intuitive reactions to danger. Risk as analysis brings logic, reason, and scientific deliberation to bear on risk management. Reliance on risk as feelings is described with "the affect heuristic." The authors trace the development of this heuristic across a variety of research paths. The authors also discuss some of the important practical implications resulting from ways that this heuristic impacts how people perceive and evaluate risk, and, more generally, how it influences all human decision making. Finally, some important implications of the affect heuristic for communication and decision making pertaining to cancer prevention and treatment are briefly discussed. PMID- 16045418 TI - Short-term cost for long-term benefit: time preference and cancer control. AB - A tradeoff between short-term costs and long-term gains characterizes many cancer control behaviors, such as behavior change (e.g., quitting smoking), screening (e.g., mammography), and prevention (e.g., healthy diet). One factor that may influence these tradeoffs is time preference, or the value assigned to future outcomes relative to immediate ones. Studies of the relationship between individual differences in time preference and preventive health behaviors, however, have yielded mixed results. Time preference is related to addictive behaviors (e.g., smoking) but not to other preventive health behaviors (e.g., vaccination). This pattern of results suggests that time preference measures reflect an ability to forgo immediate gratification that is applicable to hot behaviors, such as smoking, but not to cold behaviors, such as vaccination. PMID- 16045419 TI - Hot-cold empathy gaps and medical decision making. AB - Prior research has shown that people mispredict their own behavior and preferences across affective states. When people are in an affectively "cold" state, they fail to fully appreciate how "hot" states will affect their own preferences and behavior. When in hot states, they underestimate the influence of those states and, as a result, overestimate the stability of their current preferences. The same biases apply interpersonally; for example, people who are not affectively aroused underappreciate the impact of hot states on other people's behavior. After reviewing research documenting such intrapersonal and interpersonal hot-cold empathy gaps, this article examines their consequences for medical, and specifically cancer-related, decision making, showing, for example, that hot-cold empathy gaps can lead healthy persons to expose themselves excessively to health risks and can cause health care providers to undertreat patients for pain. PMID- 16045420 TI - Misimagining the unimaginable: the disability paradox and health care decision making. AB - Good decision making often requires accurate predictions about how potential outcomes will make one feel. However, people often mispredict the emotional impact of unfamiliar circumstances. For example, they often overestimate the emotional impact that chronic illnesses and disability will have on their lives. In the present article, the authors look at possible sources of error in both the happiness reports of patients with chronic illness or disability and the happiness predictions of healthy people asked to imagine the same illnesses and disabilities. On balance, the available evidence suggests that, whereas patients misreport their well-being, healthy people also mispredict the emotional impact that chronic illness and disability will have on their lives. PMID- 16045421 TI - Advance directives and cancer decision making near the end of life. AB - Seriously ill individuals, including those seriously ill with cancer, are frequently encouraged to complete instructional advance directives (i.e., living wills) to ensure that their wishes about the use of life-sustaining treatment are honored if they should lose the ability to make decisions for themselves. The authors present a social psychological analysis making explicit a series of steps that must necessarily take place if living wills are to honor the wishes of incapacitated patients. They then focus on 3 key steps in the analysis and review relevant research from the medical and psychological literatures. In each case, this research raises serious questions about the psychological assumptions underlying the effective use of living wills in end-of-life decision making. Discussion focuses on the need for policy and law guiding the use of advance directives to be informed by both basic and applied research on judgment and decision making. PMID- 16045422 TI - Decision counseling in cancer prevention and control. AB - Informed and shared decision making are hallmarks of quality medical care. Although decision aids (e.g., brochures, decision boards, videos, interactive computer programs) can impart useful information, there is a dearth of work on theory-based approaches that help people clarify preferences and select a favored alternative. Decision counseling is a novel method that has been developed to address this need. In a decision counseling session, provider and patient identify personal values associated with decision alternatives, weigh the influence of relevant factors, clarify preference, and select an option from available alternatives. Decision counseling is described here in relation to 3 decision-making situations (i.e., having cancer screening, being tested for cancer risk, and joining a cancer chemoprevention trial). Preliminary findings suggest that decision counseling can help to clarify personal preferences related to health behavior choices and, thus, facilitate achievement of the ideals of informed and shared decision making. PMID- 16045423 TI - Decision making and decision support for hereditary breast-ovarian cancer susceptibility. AB - Genetic testing for disease susceptibility has the potential to revolutionize health care by allowing for individually tailored disease prevention strategies. To achieve this promise, patients and physicians must use the information obtained through genetic testing to make medical decisions that are consistent with patient preferences and that lead to reduced disease morbidity and mortality. However, decisions associated with genetic testing can be complex. In this article, the authors review decision making associated with genetic testing and the medical management of hereditary breast-ovarian cancer susceptibility. They focus on decisions regarding BRCA1/2 testing and prophylactic surgery among BRCA1 and BRCA2 mutation carriers. They highlight the role of patient preferences and decision support in this population. The studies reviewed indicate that although patients' preferences do predict genetic testing and management decisions, other factors also influence their decision making. In particular, the authors discuss the role of anxiety and worry in relation to testing and surgery decisions. PMID- 16045424 TI - Patients' values and clinical substituted judgments: the case of localized prostate cancer. AB - The authors examined agreement between patients' utilities and importance rankings and clinicians' judgments of these assessments using a multiattribute model representing 6 aspects of health states potentially associated with localized prostate cancer. Patients were interviewed individually shortly after diagnosis and at a follow-up visit to obtain time-tradeoff utilities for 4 health states, including current health, and importance ranks of the 6 attributes. Their clinicians independently provided views of what utilities and importance ranks would be in the patient's best interest. Using patient-clinician pairs as the unit of analysis, the authors discovered that only about 50% of the correlations across 4 health states were high enough (.80) to be acceptable for clinical use for substituted judgment. Their conclusion: Clinicians should recognize that their judgments of the utility of health states associated with localized prostate cancer may not correspond closely with those of the patient. PMID- 16045425 TI - Decision research strategies. AB - Cancer poses many, often difficult choices. Studying these choices poses several strategic decisions for researchers, including (a) whether to conduct formal analyses of the choices being studied, (b) whether to adopt a persuasive stance (or only facilitate independent decision making), (c) whether to focus on optimizing specific choices or securing broader mastery, and (d) which individual differences to address. Behavioral decision research's strategic approach is demonstrated in 4 contexts relevant to cancer-related decisions: (a) informed consent, (b) prevention, (c) infectious disease, and (d) medical emergencies. Each example contains interacting elements of normative analysis, identifying optimal choices; descriptive research, characterizing actual behavior; and prescriptive interventions, seeking to bridge the gap between the normative ideal and the descriptive reality. PMID- 16045426 TI - A contextual approach to treatment decision making among breast cancer survivors. AB - This article presents a contextual model of adjustment to cancer that frames research inquiry on treatment decision making among long-term breast cancer survivors. Psychosocial adaptation, of which treatment decision making is a part, is viewed within a social ecological framework (T. A. Revenson, 1990, 2003) that encompasses 4 contexts: the situational context, the interpersonal context, the sociocultural context, and the temporal context. Examples of how each context may influence decision-making processes are described, and guidelines for future research are provided. PMID- 16045427 TI - A communication model of shared decision making: accounting for cancer treatment decisions. AB - The authors present a communication model of shared decision making (CMSDM) that explicitly identifies the communication process as the vehicle for decision making in cancer treatment. In this view, decision making is necessarily a sociocommunicative process whereby people enter into a relationship, exchange information, establish preferences, and choose a course of action. The model derives from contemporary notions of behavioral decision making and ethical conceptions of the doctor-patient relationship. This article briefly reviews the theoretical approaches to decision making, notes deficiencies, and embeds a more socially based process into the dynamics of the physician-patient relationship, focusing on cancer treatment decisions. In the CMSDM, decisions depend on (a) antecedent factors that have potential to influence communication, (b) jointly constructed communication climate, and (c) treatment preferences established by the physician and the patient. PMID- 16045428 TI - [Stability of thermolabile pharmaceutical specialities under varios temperature conditions]. PMID- 16045429 TI - [Minocycline-induced pleurocarditis and eosinophilic pneumonia: a propos of a case]. PMID- 16045430 TI - [Omeprazol-associated intersticial nephritis. A case report]. PMID- 16045431 TI - [Dual agranulocytosis episode likely resulting from metamizole use]. PMID- 16045432 TI - [Cytostatic drug identification using bar codes: an essential need for patient safety]. PMID- 16045433 TI - [Monitoring free valproic acid concentrations in a post-surgical patient with sepsis]. PMID- 16045434 TI - [Hypersensitivity response to cyclophosphamide in a patient with systemic lupus erythematous]. PMID- 16045439 TI - L-Alanine induces changes in metabolic and signal transduction gene expression in a clonal rat pancreatic beta-cell line and protects from pro-inflammatory cytokine-induced apoptosis. AB - Acute effects of nutrient stimuli on pancreatic beta-cell function are widely reported; however, the chronic effects of insulinotropic amino acids, such as L alanine, on pancreatic beta-cell function and integrity are unknown. In the present study, the effects of prolonged exposure (24 h) to the amino acid L alanine on insulin secretory function, gene expression and pro-inflammatory cytokine-induced apoptosis were studied using clonal BRIN-BD11 cells. Expression profiling of BRIN-BD11 cells chronically exposed to L-alanine was performed using oligonucleotide microarray analysis. The effect of alanine, the iNOS (inducible nitric oxide synthase) inhibitor NMA (N(G)-methyl-L-arginine acetate) or the iNOS and NADPH oxidase inhibitor DPI (diphenylene iodonium) on apoptosis induced by a pro-inflammatory cytokine mix [IL-1beta (interleukin-1beta), TNF-alpha (tumour necrosis factor-alpha) and IFN-gamma (interferon-gamma)] was additionally assessed by flow cytometry. Culture for 24 h with 10 mM L-alanine resulted in desensitization to the subsequent acute insulin stimulatory effects of L-alanine. This was accompanied by substantial changes in gene expression of BRIN-BD11 cells. Sixty-six genes were up-regulated >1.8-fold, including many involved in cellular signalling, metabolism, gene regulation, protein synthesis, apoptosis and the cellular stress response. Subsequent functional experiments confirmed that L-alanine provided protection of BRIN-BD11 cells from pro-inflammatory cytokine-induced apoptosis. Protection from apoptosis was mimicked by NMA or DPI suggesting L-alanine enhances intracellular antioxidant generation. These observations indicate important long-term effects of L-alanine in regulating gene expression, secretory function and the integrity of insulin-secreting cells. Specific amino acids may therefore play a key role in beta-cell function in vivo. PMID- 16045441 TI - Impaired glutamine metabolism in NMDA receptor hypofunction induced by MK801. AB - Paradoxically, glutamate receptor antagonists have neurotoxic and psychotogenic properties in addition to their neuroprotective potential during excessive glutamate release. In the present study the non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist MK801 was used to examine glial-neuronal interactions in NMDA receptor hypofunction. Rats were given a subanesthetic dose of MK801 together with [1-13C]glucose and [1,2-13C]acetate, and brains were removed 20 min later. Analyses of extracts from cingulate, retrosplenial plus middle frontal cortices (CRFC) and temporal lobe were performed using HPLC and 13C and 1H nuclear magnetic resonance spectroscopy. Hypofunction of the NMDA receptor induced similar changes in both brain areas investigated; however, the changes were most pronounced in the temporal lobe. Generally, only labeling from [1 13C]glucose was affected by MK801. In CRFC and temporal lobe amounts of both labeled and unlabeled glutamine were increased, whereas those of aspartate were decreased. In the CRFC the decrease in labeling of aspartate was greater than the decrease in concentration, leading to decreased 13C enrichment. In temporal lobe, not in CRFC, increased concentrations of glutamate, GABA, succinate, glutathione and inositol were detected together with increased labeling of GABA and succinate from [1-13C]glucose. 13C Enrichment was decreased in glutamate and increased in succinate. The results point towards a disturbance in glutamate-glutamine cycling and thus interaction between neurons and glia, since labeling of glutamate and glutamine from glucose was affected differently. PMID- 16045445 TI - Evidence for low GluR2 AMPA receptor subunit expression at synapses in the rat basolateral amygdala. AB - Fast excitatory synaptic responses in basolateral amygdala (BLA) neurons are mainly mediated by ionotropic glutamate receptors of the alpha-amino-3-hydroxy-5 methylisoxazole-4-propionate (AMPA) subtype. AMPA receptors containing an edited GluR2 subunit are calcium impermeable, whereas those that lack this subunit are calcium permeable and also inwardly rectifying. Here, we sought to determine the extent to which synapses in the rat BLA have AMPA receptors with GluR2 subunits. We assessed GluR2 protein expression in the BLA by immunocytochemistry with a GluR2 subunit-specific antiserum at the light and electron microscopic level; for comparison, a parallel examination was carried out in the hippocampus. We also recorded from amygdala brain slices to examine the voltage-dependent properties of AMPA receptor- mediated evoked synaptic currents in BLA principal neurons. At the light microscopic level, GluR2 immunoreactivity was localized to the perikarya and proximal dendrites of BLA neurons; dense labeling was also present over the pyramidal cell layer of hippocampal subfields CA1 and CA3. In electron micrographs from the BLA, most of the synapses were asymmetrical with pronounced postsynaptic densities (PSD). They contained clear, spherical vesicles apposed to the PSD and were predominantly onto spines (86%), indicating that they are mainly with BLA principal neurons. Only 11% of morphological synapses in the BLA were onto postsynaptic elements that showed GluR2 immunoreactivity, in contrast to hippocampal subfields CA1 and CA3 in which 76% and 71% of postsynaptic elements were labeled (p < 0.001). Synaptic staining in the BLA and hippocampus, when it occurred, was exclusively postsynaptic, and particularly heavy over the PSD. In whole-cell voltage clamp recordings, 72% of BLA principal neurons exhibited AMPA receptor-mediated synaptic currents evoked by external capsule stimulation that were inwardly rectifying. Although BLA principal neurons express perikaryal and proximal dendritic GluR2 immunoreactivity, few synapses onto these neurons express GluR2, and a preponderance of principal neurons have inwardly rectifying AMPA-mediated synaptic currents, suggesting that targeting of GluR2 to synapses is restricted. Many BLA synaptic AMPA receptors are likely to be calcium permeable and could play roles in synaptic plasticity, epileptogenesis and excitoxicity. PMID- 16045446 TI - The mitochondrial uncoupler 2,4-dinitrophenol attenuates tissue damage and improves mitochondrial homeostasis following transient focal cerebral ischemia. AB - Ischemic stroke is caused by acute neuronal degeneration provoked by interruption of cerebral blood flow. Although the mechanisms contributing to ischemic neuronal degeneration are myriad, mitochondrial dysfunction is now recognized as a pivotal event that can lead to either necrotic or apoptotic neuronal death. Lack of suitable 'upstream' targets to prevent loss of mitochondrial homeostasis has, so far, restricted the development of mechanistically based interventions to promote neuronal survival. Here, we show that the uncoupling agent 2,4 dinitrophenol (DNP) reduces infarct volume approximately 40% in a model of focal ischemia reperfusion injury in the rat brain. The mechanism of protection involves an early decrease in mitochondrial reactive oxygen species formation and calcium uptake leading to improved mitochondrial function and a reduction in the release of cytochrome c into the cytoplasm. The observed effects of DNP were not associated with enhanced cerebral perfusion. These findings indicate that compounds with uncoupling properties may confer neuroprotection through a mechanism involving stabilization of mitochondrial function. PMID- 16045448 TI - Identification and functional characterization of cereblon as a binding protein for large-conductance calcium-activated potassium channel in rat brain. AB - Large-conductance Ca2+-activated K+ (BK(Ca)) channels are activated by membrane depolarization and modulated by intracellular Ca2+. Here, we report the direct interaction of cereblon (CRBN) with the cytosolic carboxy-terminus of the BK(Ca) channel alpha subunit (Slo). Rat CRBN contained the N-terminal domain of the Lon protease, a 'regulators of G protein-signaling' (RGS)-like domain, a leucine zipper (LZ) motif, and four putative protein kinase C (PKC) phosphorylation sites. RNA messages of rat cereblon (rCRBN) were widely distributed in different tissues with especially high-levels of expression in the brain. Direct association of rCRBN with the BK(Ca) channel was confirmed by immunoprecipitation in brain lysate, and the two proteins were co-localized in cultured rat hippocampal neurons. Ionic currents evoked by the rSlo channel were dramatically suppressed upon coexpression of rCRBN. rCRBN decreased the formation of the tetrameric rSlo complex thus reducing the surface expression of functional channels. Therefore, we suggest that CRBN may play an important role in assembly and surface expression of functional BK(Ca) channels by direct interaction with the cytosolic C-terminus of its alpha-subunit. PMID- 16045451 TI - Effects of creatine treatment on survival and differentiation of GABA-ergic neurons in cultured striatal tissue. AB - Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder, characterized by a prominent loss of GABA-ergic medium-sized spiny neurons in the caudate putamen. There is evidence that impaired energy metabolism contributes to neuronal death in HD. Creatine is an endogenous substrate for creatine kinases and thereby supports cellular ATP levels. This study investigated the effects of creatine supplementation (5 mm) on cell survival and neuronal differentiation in striatal cultures. Chronic creatine treatment resulted in significant increased densities of GABA-immunoreactive (-ir) neurons, although total neuronal cell number and general viability were not affected. Similar effects were seen after short-term treatment, suggesting that creatine acted as a differentiation factor. Inhibitors of transcription or translation did not abolish the creatine-mediated effects, nor did omission of extracellular calcium, whereas inhibition of mitogen activated protein kinase and phosphatidylinositol-3-kinase significantly attenuated the creatine induced increase in GABA-ir cell densities. Creatine exhibited significant neuroprotection against toxicity instigated either by glucose- and serum deprivation or addition of 3-nitropropionic acid. In sum, the neuroprotective properties in combination with promotion of neuronal differentiation suggest that creatine has potential as a therapeutic drug in the treatment of neurodegenerative diseases, like HD. PMID- 16045452 TI - Two types of detergent-insoluble, glycosphingolipid/cholesterol-rich membrane domains from isolated myelin. AB - Two different types of low-density detergent-insoluble glycosphingolipid-enriched membrane domain (DIG) fractions were isolated from myelin by extraction with Triton X-100 (TX-100) in 50 mM sodium phosphate buffer at room temperature (20 degrees C) (procedure 1), in contrast to a single low-density fraction obtained by extraction with TX-100 in Tris buffer containing 150 mM NaCl and 5 mM EDTA at 4 degrees C (procedure 2). Procedure 1 has been used in the past by others for myelin extraction to preserve the cytoskeleton and/or radial component of oligodendrocytes and myelin, whereas procedure 2 is now more commonly used to isolate myelin DIG fractions. The two DIG fractions obtained by procedure 1 gave opaque bands, B1 and B2, at somewhat lower and higher sucrose density respectively than myelin itself. The single DIG fraction obtained by procedure 2 gave a single opaque band at a similar sucrose density to B1. Both B1 and B2 had characteristics of lipid rafts, i.e. high galactosylceramide and cholesterol content and enrichment in GPI-linked 120-kDa neural cell adhesion molecule (NCAM)120, as found by others for the single low-density DIG fraction obtained by procedure 2. However, B2 had most of the myelin GM1 and more of the sulfatide than B1, and they differed significantly in their protein composition. B2 contained 41% of the actin, 100% of the tubulin, and most of the flotillin-1 and caveolin in myelin, whereas B1 contained more NCAM120 and other proteins than B2. The single low-density DIG fraction obtained by procedure 2 contained only low amounts of actin and tubulin. B1 and B2 also had size-isoform selectivity for some proteins, suggesting specific interactions and different functions of the two membrane domains. We propose that B1 may come from non-caveolar raft domains whereas B2 may derive from caveolin-containing raft domains associated with cytoskeletal proteins. Some kinases present were active on myelin basic protein suggesting that the DIGs may come from signaling domains. PMID- 16045453 TI - Evidence that protein kinase Calpha interacts with and regulates the glial glutamate transporter GLT-1. AB - Many of the sodium-dependent neurotransmitter transporters are rapidly (within minutes) regulated by protein kinase C (PKC), with changes in activity being correlated with changes in transporter trafficking to or from the plasma membrane. Our recent studies suggest that one of the classical subtypes of PKC, PKCalpha, may selectively mediate redistribution of the neuronal glutamate transporter, excitatory amino acid carrier (EAAC)1, and show that PKCalpha can be co-immunoprecipitated with EAAC1. When the glial glutamate transporter GLT-1a is transfected into C6 glioma cells, this transporter is internalized in response to activation of PKC, but the PKC subtype involved in this regulation is unknown. In the present study, expression of the phorbol ester-activated subtypes of PKC was examined in C6 glioma transfected with GLT-1. Of the classical subtypes, only PKCalpha was detected, and of the non-classical subtypes, PKCdelta and PKCepsilon were detected. In this system, phorbol ester-dependent internalization of GLT-1 was blocked by a general inhibitor of PKCs (bisindolylmaleimide II) and by concentrations of Go6976 that selectively block classical PKCs, but not by an inhibitor of PKCdelta (rottlerin). PKCalpha immunoreactivity was found in GLT-1 immunoprecipitates obtained from transfected C6 cells and from crude rat brain synaptosomes, a milieu that better mimics in vivo conditions. The amount of PKCalpha in both types of immunoprecipitate was modestly increased by phorbol ester, and this increase was blocked by a PKC antagonist. These studies suggest that PKCalpha may be required for the regulated redistribution of GLT-1. PMID- 16045454 TI - Decreases in phosphoinositide-3-kinase/Akt and extracellular signal-regulated kinase 1/2 signaling activate components of spinal motoneuron death. AB - Motoneuron dependence on target-derived trophic factors during development is well established, with loss of trophic support leading to the death of these cells. A complete understanding of the intracellular signal transduction machinery associated with extracellular survival signals requires the examination of individual pathways in various cellular and environmental contexts. In cells deprived of trophic support, and hence compromised for survival, phosphoinositide 3-kinase (PI3K) is decreased when compared with healthy cells supplied with trophic support. Extracellular signal-regulated kinase 1/2 (ERK1/2) signaling is dramatically decreased in deprived cells. We have examined the role of these two pathways to understand how changes in their activity regulate motoneuron survival and death. Pharmacological inhibition of PI3K attenuated motoneuron survival and was important in the regulation of Bcl-2 serine phosphorylation, limited release of cytochrome c into the cytoplasm and caspase activation. Bax translocation from cytoplasm to mitochondria was not altered when PI3K was inhibited. High levels of ERK1/2 inhibition robustly attenuated motoneuron survival in cells supplied with trophic support, whereas moderate inhibition of ERK1/2 activation had little effect. ERK1/2 inhibition in these cells decreased Bcl-2 phosphorylation and resulted in release of cytochrome c from the mitochondria. Bax translocation and caspase activation were not affected by ERK1/2 inhibition. These data reveal that changes in PI3K and ERK1/2 signaling lead to individual and overlapping effects on the cell-death machinery. Characterizing the role of these pathways is critical for a fundamental understanding of the development and degeneration of specific neuronal populations. PMID- 16045455 TI - Structural biology of NCAM homophilic binding and activation of FGFR. AB - In this review, we analyse the structural basis of the homophilic interactions of the neural cell adhesion molecule (NCAM) and the NCAM-mediated activation of the fibroblast growth factor receptor (FGFR). Recent structural evidence suggests that NCAM molecules form cis-dimers in the cell membrane through a high affinity interaction. These cis-dimers, in turn, mediate low affinity trans-interactions between cells via formation of either one- or two-dimensional 'zippers'. We provide evidence that FGFR is probably activated by NCAM very differently from the way by which it is activated by FGFs, reflecting the different conditions for NCAM-FGFR and FGF-FGFR interactions. The affinity of FGF for FGFR is approximately 10(6) times higher than that of NCAM for FGFR. Moreover, in the brain NCAM is constantly present on the cell surface in a concentration of about 50 microm, whereas FGFs only appear transiently in the extracellular environment and in concentrations in the nanomolar range. We discuss the structural basis for the regulation of NCAM-FGFR interactions by two molecular 'switches', polysialic acid (PSA) and adenosine triphosphate (ATP), which determine whether NCAM acts as a signalling or an adhesion molecule. PMID- 16045456 TI - Pyoderma, group A streptococci and parasitic skin diseases -- a dangerous relationship. PMID- 16045457 TI - Molecular epidemiology of drug resistance markers of Plasmodium falciparum malaria in Thailand. AB - To determine differences in the distribution of drug resistance mutations in the Plasmodium falciparum chloroquine resistance transporter (pfcrt) and P. falciparum multi-drug resistance 1 (pfmdr1) genes of P. falciparum isolates in Thailand, a study was conducted using polymerase chain reaction-restriction fragment length polymorphism to detect mutations in P. falciparum isolates obtained from three areas with different levels of in vivo mefloquine (MQ) resistance. All isolates carried mutant allele T76 of the pfcrt gene and wild type allele D1246 of the pfmdr1 gene except for one isolate, which showed the wild-type K76 allele. This isolate was obtained from Chanthaburi Province, an area with high MQ resistance. Relatively low rates of the mutant alleles D1042 and Y86 of the pfmdr1 gene were found among Thai isolates of P. falciparum. However, a statistically significant difference in the distribution was noted. Most of the mutant isolates were found among isolates from areas with moderate or low MQ resistance. Only one isolate with mixed mutant and wild-type N1042 and D1042 and two mutants of Y86 were found among the isolates from areas with high MQ resistance. The findings provide limited support for the hypothesis that mutant alleles of pfmdr1 may be associated with increased sensitivity to MQ. PMID- 16045458 TI - Efficacy and safety of two dosages of cotrimoxazole as preventive treatment for HIV-infected Malawian adults with new smear-positive tuberculosis. AB - OBJECTIVE: To assess the efficacy and safety of two different dosages of cotrimoxazole (CTX) in prophylaxis in HIV-positive new smear-positive pulmonary tuberculosis (TB) patients in Blantyre, Malawi. METHOD: Randomized, double-blind trial using 480 and 960 mg of CTX given to new TB patients, who were followed up until the end of the tuberculosis treatment. The primary outcome was survival. The outcome in the two groups was also compared with an unselected cohort of similar patients registered in Zomba, Malawi in 1995 and new smear-positive patients registered in the National Tuberculosis Programme in 1999. The secondary outcome was the occurrence of (opportunistic) events, especially bacterial pneumonia. RESULTS: There were no statistically significant differences in mortality and bacterial pneumonia between the groups receiving the two different dosages. The case fatality rate at the end of the tuberculosis treatment was 15.4% in the 480 mg group and 14.0% in the 960 mg group. This was lower than the case fatality rate in the Zomba cohort (19.2%, P = 0.10) and lower than the case fatality rate in the national programme (21.0%, P < 0.001). CTX was well tolerated. Compliance was fair. CONCLUSIONS: CTX prophylaxis may have a beneficial effect on mortality and morbidity in HIV-infected smear-positive tuberculosis patients in Malawi. The efficacy of both dosages is not significantly different. The intervention is cheap and easy to implement. These results would support implementation of CTX in this patient group until better strategies are available or evidence is convincingly presented to suggest that its benefit is marginal. PMID- 16045459 TI - Tuberculosis and HIV interaction in sub-Saharan Africa: impact on patients and programmes; implications for policies. AB - Sub-Saharan Africa carries the overwhelming share of the global burden of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) and of HIV associated tuberculosis (TB). The impact of HIV on TB patients and programmes has implications for TB control policies. The impact on patients includes the effect of HIV on diagnosis and on the patterns of HIV-related TB, the response of HIV infected TB patients to TB treatment, the benefits of antiretroviral therapy (ART), and the quality and continuity of care for TB patients. The impact on national TB programmes (NTPs) includes increased case load, impaired NTP performance, increased need for access to ART and difficulties in reaching TB control targets. Implications for policies include the need to promote TB and HIV/AIDS programme collaboration, aimed at improving NTP performance (TB case finding and treatment outcomes), quality and continuity of care, and monitoring and interpretation of progress towards TB control targets. In order to provide the recommended international standard of care for TB patients, clinicians need to be aware of the impact of HIV on TB patients and programmes and the implications for the policies that provide the framework for this standard. Conversely, policy-makers need to understand the impact of HIV on TB patients and programmes. This can help to ensure a firm evidence base for TB control policies aiming at the high standard of patient care that is at the heart of TB and HIV programmes. PMID- 16045460 TI - No evidence of prolonged enterovirus excretion in HIV-seropositive patients. AB - Mutations frequently occur in oral poliovirus vaccine (OPV) strains upon replication in the human intestine. These strains occasionally revert to being neurovirulent. The more prolonged the excretion of OPV, the higher the risk of reversion. OPV strains can be secreted for several months in humans presenting humoral immune system deficiencies. The duration of excretion of OPV strains or other enteroviruses in individuals infected with the human immunodeficiency virus (HIV) is unknown. We investigated whether HIV infection, which is very prevalent in the Central African Republic, causes prolonged excretion of enteroviruses and, in particular, of OPV strains in adults. We studied 28 HIV-infected adults living with children who were immunized with OPV during national immunization days (NIDs). Blood samples were collected to confirm HIV status and to evaluate immunodeficiency before the NIDs. Stool samples for enterovirus isolation were also collected before the NIDs, between the two rounds of immunization and 2, 4 and 6 months after the second round of immunization. No poliovirus was isolated from any stool sample. Eight enteroviruses were isolated from eight adults (maximum one strain per patient). Enteroviruses were not more frequently isolated from severely immunodeficient patients. Thus, HIV-infected adults do not appear to be at high risk of infection with OPV strains and the excretion of enteroviruses (and thus of polioviruses) does not seem to be prolonged in HIV infected adults. PMID- 16045461 TI - Impact of vector control on a dengue fever outbreak in Trinidad, West Indies, in 1998. AB - In 1998, Trinidad experienced its first major outbreak of dengue haemorrhagic fever. Data from the Trinidad Public Health Laboratory, the National Surveillance Unit and Insect Vector Control Division, Ministry of Health, Trinidad and Tobago were analysed to determine the impact of vector control measures on the dengue outbreak. Geographical Information Systems (GIS)/Global Positioning Systems (GPS) were used to map cases and to distinguish epidemiological clusters. The Aedes aegypti population densities were higher than the 5% transmission threshold in all counties. The spatial distribution of dengue fever cases was significantly correlated with the heavily populated east-west corridor in the north and several distinctly separate clusters in the western part of the island. The temporal distribution patterns showed significantly more dengue fever cases occurring during the rainy season than during the dry season. This study documents the importance of vector control in the prevention of dengue transmission since no vaccine is currently available, and emphasizes the urgent need to understand better the environmental factors which contribute to the proliferation of this disease vector Ae. aegypti. PMID- 16045462 TI - Effectiveness of dengue control practices in household water containers in Northeast Thailand. AB - OBJECTIVE: To investigate the influence of larval control methods (using temephos, keeping fish and covering containers with lids), water use and weekly cleaning of containers on the presence of Aedes aegypti larvae in water-storage containers in rural and urban households in Khon Kaen province. METHOD: Cross sectional questionnaire survey and larval survey covered 966 households and 5821 containers were inspected. RESULT: In rural and urban areas larval control was patchy and often ineffective. Consequently, the mosquito indices exceed the target indices for dengue control with the Breteau Indices of 201 and 113, and Container Indices of 25 and 28 in rural and urban areas, respectively. The containers most frequently infested with larvae were rectangular cement containers storing water for bathing (rural: 37.2%; urban: 35%) and flushing the toilets (rural: 35.7%; urban: 34.3%). Keeping fish [adjusted odds ratio (AOR): 0.08-0.16] was the most effective methods of control. Correctly covering containers with lids was similarly effective (AOR: 0.10-0.25) when used on jars for storing drinking water. However, frequent use of containers reduced the effectiveness of lids. Temephos was effective only in dragon jars in urban areas (AOR: 0.46) where a standard package of temephos were available. Weekly cleaning of containers was an effective method for larval control in most types of containers. A combination of control methods increased effectiveness. CONCLUSION: This study highlights the complex interaction of household water use and larval control practices as well as the importance of determining the most effective control measures compatible with water practices for implementing control promotion. PMID- 16045463 TI - Participatory evaluations of trachoma control programmes in eight countries. AB - OBJECTIVES: Trachoma is a major cause of blindness. The objective of this initiative was to conduct participatory process evaluations of the trachoma control programmes receiving support from the International Trachoma Initiative in eight countries. METHODS: During each 2- to 4-week evaluation we analysed information collected at the central, district and community level through interviews, focus groups, questionnaires, direct observation of trachoma control activities, and existing data. RESULTS: Mapping and assessment of disease prevalence had been completed in four of eight countries. Integration of trachoma control activities into national planning and district-level service provision varied. Intersectoral partnerships to implement the SAFE strategy (i.e. surgery, antibiotics, facial cleanliness and environmental change) were well established in a few countries. In all eight countries, the number of surgeries performed annually was insufficient; and quality of surgery was rarely monitored. Mass distribution of antibiotics was carried out well in extremely resource-poor settings and good coverage was achieved, although the strategy for antibiotic distribution varied. Inadequate water and sanitation remained a major problem in all programme areas. Monitoring of programme activities was generally inadequate. The Morocco programme is an example from which lessons and processes can be learnt and adapted to other programme countries. CONCLUSIONS: Significant achievements have been made in implementing the SAFE strategy. Scaling up of activities to true national coverage should be planned and implemented provided the resources can be made available. Further standardization of how to assess, implement and monitor trachoma control activities will facilitate expansion of the programme. PMID- 16045464 TI - Needle stick injuries among nurses in sub-Saharan Africa. AB - OBJECTIVES: Despite a heavy burden of HIV/AIDS and other blood borne infections, few studies have investigated needle stick injuries in sub-Saharan Africa. We conducted a cross-sectional study at Mulago national referral hospital in Kampala, Uganda, to assess the occurrence and risk factors of needle stick injuries among nurses and midwives. METHODS: A total of 526 nurses and midwives involved in the direct day-to-day management of patients answered a questionnaire inquiring about occurrence of needle stick injuries and about potential predictors, including work experience, work load, working habits, training, and risk behaviour. RESULTS: A 57% of the nurses and midwives had experienced at least one needle stick injury in the last year. Only 18% had not experienced any such injury in their entire career. The rate of needle stick injuries was 4.2 per person-year. Multiple logistic regression analysis showed that the most important risk factor for needle stick injuries was lack of training on such injuries (OR 5.72, 95% CI 3.41-9.62). Other important risk factors included working for more than 40 h/week (OR 1.90, 95% CI 1.20-3.31), recapping needles most of the time (OR 1.78, 95% CI 1.11-2.86), and not using gloves when handling needles (OR 1.91, 95% CI 1.10-3.32). CONCLUSIONS: The study showed a high rate of needle stick injuries among nurses and midwives working in Uganda. The strongest predictor for needle stick injuries was lack of training. Other important risk factors were related to long working hours, working habits, and experience. PMID- 16045465 TI - Effects of vitamin A supplementation on child mortality: evidence from Nepal's 2001 Demographic and Health Survey. AB - OBJECTIVE: To assess the effect of Nepal's vitamin A supplementation programme on child mortality at age 12-59 months. MATERIALS AND METHODS: Logistic regression, applied to retrospective data from Nepal's 2001 Demographic and Health Survey. RESULTS: After a number of potentially confounding variables are controlled, the effect of 100% community-level vitamin A coverage since the child's birth, relative to no coverage, is to reduce the odds of dying at age 12-59 months by slightly more than half (OR = 0.47, P = 0.03). CONCLUSIONS: The estimated beneficial effect of vitamin A supplementation on child mortality is larger than that found in most earlier clinical studies. This larger effect may be due mainly to the other health-related activities undertaken by the female community health volunteers who distribute vitamin A capsules. PMID- 16045466 TI - The epidemiology of animal bite injuries in Uganda and projections of the burden of rabies. AB - OBJECTIVES: Rabies is a global problem, although it is often under-reported in developing countries. We aimed at describing the profile of patients presenting to health centres with animal bite injuries in Uganda, and use a predictive model to estimate the mortality of rabies at a national level. METHODS: We conducted a passive surveillance study in Uganda based in a random sample of health centres supplied with rabies vaccine to determine the characteristics of bite injury patients and establish the age and sex profiles of patients, the site of bites and their severity, wound management techniques and details of the vaccination course given. We also applied a decision tree model to the data to estimate the rabies mortality from the bite injury data using an established protocol. RESULTS: We found that most patients are bitten by dogs, and that a considerable proportion of these are young children, who are at greater risk of developing rabies in the absence of treatment due to the location of the bites they receive. From conservative parameter estimates, we estimate that in the absence of post exposure prophylaxis (PET), 592 (95% CI 345-920) deaths would occur, and that if one dose of PET is sufficient for protection following a rabid animal bite, 20 (95% CI 5-50) deaths would occur annually. If a complete course of PET is required for protection following a rabid animal bite, up to 210 (95% CI 115-359) deaths would occur, as 41% of patients did not complete their course of PET. CONCLUSIONS: Active animal bite surveillance studies are required to improve our mortality estimates and determine the true burden of rabies in the Ugandan population. We emphasize the need for small-scale active case detection studies and improved data on the recognition of rabies in dogs as inputs for improving national-level estimates of rabies mortality. PMID- 16045467 TI - Community-based health insurance in developing countries: a study of its contribution to the performance of health financing systems. AB - We studied the potential of community-based health insurance (CHI) to contribute to the performance of health financing systems. The international empirical evidence is analysed on the basis of the three health financing subfunctions as outlined in the World Health Report 2000: revenue collection, pooling of resources and purchasing of services. The evidence indicates that achievements of CHI in each of these subfunctions so far have been modest, although many CHI schemes still are relatively young and would need more time to develop. We present an overview of the main factors influencing the performance of CHI on these financing subfunctions and discuss a set of proposals to increase CHI performance. The proposals pertain to the demand for and the supply of health care in the community; to the technical, managerial and institutional set-up of CHI; and to the rational use of subsidies. PMID- 16045468 TI - Networking in international health: introducing the Federation of European Societies for Tropical Medicine and International Health (FESTMIH). PMID- 16045469 TI - The Arabidopsis ROCK-N-ROLLERS gene encodes a homolog of the yeast ATP-dependent DNA helicase MER3 and is required for normal meiotic crossover formation. AB - Recent studies in Saccharomyces cerevisiae have unveiled that meiotic recombination crossovers are formed by two genetically distinct pathways: a major interference-sensitive pathway and a minor interference-insensitive pathway. Several proteins, including the MSH4/MSH5 heterodimer and the MER3 DNA helicase, are indispensable for the interference-sensitive pathway. MSH4 homologs have been identified in mice and Arabidopsis and shown to be required for normal levels of crossovers, suggesting that the function of MSH4 may be conserved among major eukaryotic kingdoms. However, it is not known whether an MER3-like function is also required for meiosis in animals and plants. We have identified an Arabidopsis gene that encodes a putative MER3 homolog and is preferentially expressed in meiocytes. T-DNA insertional mutants of this gene exhibit defects in fertility and meiosis. Detailed cytological studies indicate that the mutants are defective in homolog synapsis and crossover formation, resulting in a reduction of bivalents and in the formation of univalents at late prophase I. We have named this gene ROCK-N-ROLLERS (RCK) to reflect the mutant phenotype of chromosomes undergoing the meiotic 'dance' either in pairs or individually. Our results demonstrate that an MER3-like function is required for meiotic crossover in plants and provide further support for the idea that Arabidopsis, like the budding yeast, possesses both interference-sensitive and insensitive pathways for crossover formation. PMID- 16045470 TI - Alteration of TGA factor activity in rice results in enhanced tolerance to Xanthomonas oryzae pv. oryzae. AB - In dicotyledonous plants broad-spectrum resistance to pathogens is established after the induction of the systemic acquired resistance (SAR) response. In Arabidopsis the NPR1 protein can regulate SAR by interacting with members of the TGA class of basic, leucine-zipper transcription factors to alter pathogenesis related (PR) gene expression. Overexpression of (At)NPR1 in Arabidopsis enhances resistance to multiple pathogens. Similarly, overexpression of (At)NPR1 in rice enhances resistance to the bacterial pathogen, Xanthomonas oryzae pv. oryzae (Xoo). These results suggest that components of the (At)NPR1-mediated SAR defense response may be conserved between monocots and dicots. To determine whether or not rice TGA factors are involved in disease resistance responses, the effect of altering the function of rice TGA2.1 was analyzed in transgenic plants. Transgenic rice overexpressing an rTGA2.1 mutant, that can no longer bind DNA, and transgenic rice that have the endogenous rTGA2.1 silenced by dsRNA-mediated silencing were generated. Both types of transgenic rice displayed increased tolerance to Xoo, were dwarfed, and had altered accumulation of PR genes. The results presented in this study suggest that wild-type rTGA2.1 has primarily a negative role in rice basal defense responses to bacterial pathogens. PMID- 16045471 TI - Kanamycin reveals the role played by glutamate receptors in shaping plant resource allocation. AB - Ionotropic glutamate receptors (iGluRs) play important roles in neurotransmission in animals. There is growing evidence that iGluRs also play important roles in plants. Using a chemical genetics approach, which combined a pH-homeostasis mutant of Arabidopsis thaliana (de-etiolated3), several different iGluR agonists, molecular modelling, and reporter gene expression in transgenic plants, we provide evidence that iGluR agonism can induce dramatic changes in plant development and metabolism. Systematic hypothesis testing revealed a signalling circuit that integrates amino acid and sugar signals to affect elongation growth and the deposition of carbon into starch and lignins. The data show that aminoglycoside antibiotics, such as kanamycin, and polyamines impinge upon this circuit. These findings provide a mechanism for the conversion of amino acid and sugar signals into an appropriate response at the gene expression level, and underline the similarities in iGluR agonism between animals and plants. PMID- 16045472 TI - FHL is required for full phytochrome A signaling and shares overlapping functions with FHY1. AB - Phytochrome A (phyA) plays a primary role in initiating seedling de-etiolation and is the only plant photoreceptor known to be activated by far-red light (FR). The signaling intermediate FHY1 appears to either participate directly in relaying the phyA signal or to positively regulate a critical signaling event(s) downstream of phyA activation. Here we identify a homolog of FHY1 named FHL (FHY1 like) as a novel signaling factor essential for complete responsiveness to phyA. FHL possesses functional nuclear localization and nuclear export signals. Lines in which FHL function was abolished by insertional mutagenesis or attenuated by RNAi-mediated suppression displayed a weaker hyposensitivity to continuous FR than fhy1 null mutants and most reported phyA signaling mutants. However, hypocotyl elongation assays indicated that suppression of FHL expression in fhy1 3 caused an insensitivity of hypocotyl elongation to FR and blue light (B) indistinguishable from that seen in phyA. Real-time PCR indicates that in FR, FHY1 transcripts are approximately 15-fold more abundant than FHL transcripts. Although both FHY1 and FHL are capable of homo- and hetero-interaction via their C-termini, the ability of FHL overexpression to restore wild-type (WT) morphological and molecular phenotypes to fhy1-3 seedlings suggests that the extreme insensitivity to FR associated with suppression of FHL expression in fhy1 3 cannot be accounted for by a critical role for FHY1-FHL heterodimers in phyA signal transmission. Rather, we suggest that the relative abundances of FHY1 and FHL in WT plants account for the differences in the severity of fhy1 and fhl mutations. As for FHY1, FHL transcript accumulation is dependent on FHY3 and is decreased after exposure to FR, R or B light. These findings reiterate the prevalence of partial degeneracy in plant signaling networks that regulate responses crucial to survival. PMID- 16045473 TI - Characterization of a novel temperature-sensitive allele of the CUL1/AXR6 subunit of SCF ubiquitin-ligases. AB - Selective protein degradation by the ubiquitin-proteasome pathway has emerged as a key regulatory mechanism in a wide variety of cellular processes. The selective components of this pathway are the E3 ubiquitin-ligases which act downstream of the ubiquitin-activating and -conjugating enzymes to identify specific substrates for ubiquitinylation. SCF-type ubiquitin-ligases are the most abundant class of E3 enzymes in Arabidopsis. In a genetic screen for enhancers of the tir1-1 auxin response defect, we identified eta1/axr6-3, a recessive and temperature-sensitive mutation in the CUL1 core component of the SCF(TIR1) complex. The axr6-3 mutation interferes with Skp1 binding, thus preventing SCF complex assembly. axr6-3 displays a pleiotropic phenotype with defects in numerous SCF-regulated pathways including auxin signaling, jasmonate signaling, flower development, and photomorphogenesis. We used axr6-3 as a tool for identifying pathways likely to be regulated by SCF-mediated proteolysis and propose new roles for SCF regulation of the far-red light/phyA and sugar signaling pathways. The recessive inheritance and the temperature-sensitive nature of the pleiotropically acting axr6-3 mutation opens promising possibilities for the identification and investigation of SCF-regulated pathways in Arabidopsis. PMID- 16045474 TI - Cell adhesion in Arabidopsis thaliana is mediated by ECTOPICALLY PARTING CELLS 1- a glycosyltransferase (GT64) related to the animal exostosins. AB - Despite the fact that several hundred glycosyltransferases have been identified from sequencing of plant genomes, the biological functions of only a handful have been established to date. A Poplar glycosyltransferase 64 (GT64) family member that is differentially expressed during the cell division and elongation phases of cambial growth was identified from previously generated transcript profiling of cambium tissues. The predicted Poplar GT64 protein has a closely related Arabidopsis homolog ECTOPICALLY PARTING CELLS (EPC1). Mutation of the EPC1 gene, one of three Arabidopsis GT64 family members, results in plants with a dramatically reduced growth habit, defects in vascular formation and reduced cell cell adhesion properties in hypocotyl and cotyledon tissues. Secondary growth is enhanced in epc1 hypocotyl tissues and it is proposed that this results from the abnormal cell-cell adhesion within the cortical parenchyma cell layers. Loss of cell-cell contacts within cotyledon and leaf tissues is also proposed to account for vascular patterning defects and the fragile nature of epc1 tissues. The EPC1 protein thus plays a critical role during plant development in maintaining the integrity of organs via cell-cell adhesion, thereby providing mechanical strength and facilitating the movement of metabolites throughout the plant. PMID- 16045475 TI - Soluble starch synthase I: a major determinant for the synthesis of amylopectin in Arabidopsis thaliana leaves. AB - A minimum of four soluble starch synthase families have been documented in all starch-storing green plants. These activities are involved in amylopectin synthesis and are extremely well conserved throughout the plant kingdom. Mutants or transgenic plants defective for SSII and SSIII isoforms have been previously shown to have a large and specific impact on the synthesis of amylopectin while the function of the SSI type of enzymes has remained elusive. We report here that Arabidopsis mutants, lacking a plastidial starch synthase isoform belonging to the SSI family, display a major and novel type of structural alteration within their amylopectin. Comparative analysis of beta-limit dextrins for both wild type and mutant amylopectins suggests a specific and crucial function of SSI during the synthesis of transient starch in Arabidopsis leaves. Considering our own characterization of SSI activity and the previously described kinetic properties of maize SSI, our results suggest that the function of SSI is mainly involved in the synthesis of small outer chains during amylopectin cluster synthesis. PMID- 16045476 TI - Molecular tailoring of farnesylation for plant drought tolerance and yield protection. AB - Protecting crop yield under drought stress is a major challenge for modern agriculture. One biotechnological target for improving plant drought tolerance is the genetic manipulation of the stress response to the hormone abscisic acid (ABA). Previous genetic studies have implicated the involvement of the beta subunit of Arabidopsis farnesyltransferase (ERA1) in the regulation of ABA sensing and drought tolerance. Here we show that molecular manipulation of protein farnesylation in Arabidopsis, through downregulation of either the alpha- or beta-subunit of farnesyltransferase enhances the plant's response to ABA and drought tolerance. To test the effectiveness of tailoring farnesylation in a crop plant, transgenic Brassica napus carrying an ERA1 antisense construct driven by a drought-inducible rd29A promoter was examined. In comparison with the non transgenic control, transgenic canola showed enhanced ABA sensitivity, as well as significant reduction in stomatal conductance and water transpiration under drought stress conditions. The antisense downregulation of canola farnesyltransferase for drought tolerance is a conditional and reversible process, which depends on the amount of available water in the soil. Furthermore, transgenic plants were more resistant to water deficit-induced seed abortion during flowering. Results from three consecutive years of field trial studies suggest that with adequate water, transgenic canola plants produced the same amount of seed as the parental control. However, under moderate drought stress conditions at flowering, the seed yields of transgenic canola were significantly higher than the control. Using protein farnesyltransferase as an effective target, these results represent a successful demonstration of engineered drought tolerance and yield protection in a crop plant under laboratory and field conditions. PMID- 16045477 TI - Overexpression of ADC2 in Arabidopsis induces dwarfism and late-flowering through GA deficiency. AB - We have obtained Arabidopsis thaliana transgenic plants constitutively overexpressing ADC2, one of the two genes encoding arginine decarboxylase (ADC) in Arabidopsis. These plants contained very high levels of putrescine (Put) but no changes were observed in spermidine and spermine contents. The results obtained from quantification of free and conjugated polyamines suggest that conjugation may be a limiting step for control of Put homeostasis within a non toxic range for plant survival. Transgenic plants with increased levels of ADC2 transcript and elevated Put content showed dwarfism and late-flowering, and the phenotype was rescued by gibberellin A3 (GA3) application. The contents of bioactive GA4 and GA1, and of GA9 (a precursor of GA4), as well as the levels of AtGA20ox1, AtGA3ox1 and AtGA3ox3 transcripts (quantified by real-time PCR) were lower in the ADC2 overexpressor plants than in the wild type. No change in the expression of genes encoding earlier enzymes in the GA biosynthesis pathway was detected by microarray analysis. These results suggest that Put accumulation affects GA metabolism through the repression of biosynthetic steps catalyzed by GA 20-oxidase and GA 3-oxidase. PMID- 16045478 TI - Arabidopsis CUL3A and CUL3B genes are essential for normal embryogenesis. AB - Cullin (CUL)-dependent ubiquitin ligases form a class of structurally related multisubunit enzymes that control the rapid and selective degradation of important regulatory proteins involved in cell cycle progression and development, among others. The CUL3-BTB ligases belong to this class of enzymes and despite recent findings on their molecular composition, our knowledge on their functions and substrates remains still very limited. In contrast to budding and fission yeast, CUL3 is an essential gene in metazoans. The model plant Arabidopsis thaliana encodes two related CUL3 genes, called CUL3A and CUL3B. We recently reported that cul3a loss-of-function mutants are viable but exhibit a mild flowering and light sensitivity phenotype. We investigated the spatial and temporal expression of the two CUL3 genes in reproductive tissues and found that their expression patterns are largely overlapping suggesting possible functional redundancy. Thus, we investigated the consequences on plant development of combined Arabidopsis cul3a cul3b loss-of-function mutations. Homozygous cul3b mutant plants developed normally and were fully fertile. However, the disruption of both the CUL3A and CUL3B genes reduced gametophytic transmission and caused embryo lethality. The observed embryo abortion was found to be under maternal control. Arrest of embryogenesis occurred at multiple stages of embryo development, but predominantly at the heart stage. At the cytological level, CUL3 loss-of-function mutations affected both embryo pattern formation and endosperm development. PMID- 16045479 TI - Ex vitro composite plants: an inexpensive, rapid method for root biology. AB - Plant transformation technology is frequently the rate-limiting step in gene function analysis in non-model plants. An important tool for root biologists is the Agrobacterium rhizogenes-derived composite plant, which has made possible genetic analyses in a wide variety of transformation recalcitrant dicotyledonous plants. The novel, rapid and inexpensive ex vitro method for producing composite plants described in this report represents a significant advance over existing composite plant induction protocols, which rely on expensive and time-consuming in vitro conditions. The utility of the new system is validated by expression and RNAi silencing of GFP in transgenic roots of composite plants, and is bolstered further by experimental disruption, via RNAi silencing, of endogenous plant resistance to the plant parasitic nematode Meloidogyne incognita in transgenic roots of Lycopersicon esculentum cv. Motelle composite plants. Critical parameters of the method are described and discussed herein. PMID- 16045480 TI - A new seed-based assay for meiotic recombination in Arabidopsis thaliana. AB - Meiotic recombination is a fundamental biological process that plays a central role in the evolution and breeding of plants. We have developed a new seed-based assay for meiotic recombination in Arabidopsis. The assay is based on the transformation of green and red fluorescent markers expressed under a seed specific promoter. A total of 74 T-DNA markers were isolated, sequenced and mapped both physically and genetically. Lines containing red and green markers that map 1-20 cM apart were crossed to produce tester lines with the two markers linked in cis yielding seeds that fluoresced both in red and green. We show that these lines can be used for efficient scoring of recombinant types (red only or green only fluorescing seeds) in a seed population derived from a test cross (backcross) or self-pollination. Two tester lines that were characterized during several generations of backcross and self-pollination, one in the background of ecotype Landsberg and one in the ecotype Columbia, are described. We discuss the number of plants and seeds to be scored in order to obtain reliable and reproducible crossing over rate values. This assay offers a relatively high throughput method, with the benefit of seed markers (similar to the maize classical genetic markers) combined with the advantages of Arabidopsis. It advances the prospect to better understand the factors that affect the rate of meiotic crossover in plants and to stimulate this process for more efficient breeding and mapping. PMID- 16045482 TI - GABA and glycine are protective to mature but toxic to immature rat cortical neurons under hypoxia. AB - Although recent studies suggest that gamma-aminobutyric acid (GABA) and glycine may be 'inhibitory' to mature neurons, but 'excitatory' to immature neurons under normoxia, it is unknown whether inhibitory neurotransmitters are differentially involved in neuronal response to hypoxia in immature and mature neurons. In the present study, we exposed rat cortical neurons to hypoxia (1% O2) and examined the effects of three major inhibitory neurotransmitters (GABA, glycine and taurine) on the hypoxic neurons at different neuronal ages [days in vitro (DIV)4 20]. Our data showed that the cortical neurons expressed both GABA(A) and glycine receptors with differential developmental profiles. GABA (10-2000 microm) was neuroprotective to hypoxic neurons of DIV20, but enhanced hypoxic injury in neurons of DIV4. In comparison with delta-opioid receptor (DOR)-induced protection in DIV20 neurons exposed to 72 h of hypoxia, glycine-induced protection was weaker than that of DOR but stronger than that of GABA and taurine. These data suggest that the effects of the inhibitory neurotransmitters on hypoxic cortical neurons are age dependent, with GABA and glycine being neurotoxic to immature neurons and neuroprotective to mature neurons. PMID- 16045483 TI - Role of GSK-3beta activity in motor neuronal cell death induced by G93A or A4V mutant hSOD1 gene. AB - Point mutations such as G93A and A4V in the human Cu/Zn-superoxide dismutase gene (hSOD1) cause familial amyotrophic lateral sclerosis (fALS). In spite of several theories to explain the pathogenic mechanisms, the mechanism remains largely unclear. Increased activity of glycogen synthase kinase-3 (GSK-3) has recently been emphasized as an important pathogenic mechanism of neurodegenerative diseases, including Alzheimer's disease and ALS. To investigate the effects of G93A or A4V mutations on the phosphatidylinositol-3-kinase (PI3-K)/Akt and GSK-3 pathway as well as the caspase-3 pathway, VSC4.1 motoneuron cells were transfected with G93A- or A4V-mutant types of hSOD1 (G93A and A4V cells, respectively) and, 24 h after neuronal differentiation, their viability and intracellular signals, including PI3-K/Akt, GSK-3, heat shock transcription factor-1 (HSTF-1), cytochrome c, caspase-3 and poly(ADP-ribose) polymerase (PARP), were compared with those of wild type (wild cells). Furthermore, to elucidate the role of the GSK-3beta-mediated cell death mechanism, alterations of viability and intracellular signals in those mutant motoneurons were investigated after treating the cells with GSK-3beta inhibitor. Compared with wild cells, viability was greatly reduced in the G93A and A4V cells. However, the treatment of G93A and A4V cells with GSK-3beta inhibitor increased their viability by activating HSTF-1 and by reducing cytochrome c release, caspase-3 activation and PARP cleavage. However, the treatment did not affect the expression of PI3-K/Akt and GSK-3beta. These results suggest that the G93A or A4V mutations inhibit PI3 K/Akt and activate GSK-3beta and caspase-3, thus becoming vulnerable to oxidative stress, and that the GSK-3beta-mediated cell death mechanism is important in G93A and A4V cell death. PMID- 16045484 TI - Selective sparing of hippocampal CA3 cells following in vitro ischemia is due to selective inhibition by acidosis. AB - A brief global ischemic insult to the brain leads to a selective degeneration of the pyramidal neurons in the hippocampal CA1 region while the neurons in the neighbouring CA3 region are spared. The reason for this difference is not known. The selective vulnerability of CA1 neurons to ischemia can be reproduced in vitro in murine organotypic slice cultures, if the ion concentrations in the medium during the anoxic/aglycemic insult are similar to that in the brain extracellular fluid during ischemia in vivo. As acidosis develops during ischemia, we studied the importance of extracellular pH for selective vulnerability. We found that cell death in the CA1 and CA3 regions was equally prevented by removal of calcium from the medium or following blockade of the N-methyl-D-aspartate (NMDA) receptor by D-2 amino-5-phosphonopentanoic-acid (D-APV). On the other hand, damage to the CA3 neurons markedly decreased with decreasing pH following in vitro ischemia, while the degeneration of CA1 neurons was less pH dependent. Patch-clamp recordings from pyramidal neurons in the CA1 and CA3 regions, respectively, revealed a pronounced inhibition of NMDA-receptor mediated excitatory postsynaptic currents (EPSCs) at pH 6.5 that was equally pronounced in the two regions. However, when changing pH from 6.5 to 7.4 the recovery of the EPSCs was significantly slower in the CA3 region. We conclude that acidosis selectively protects CA3 pyramidal neurons during in vitro ischemia, and differentially affects the kinetics of NMDA receptor activation, which may explain the difference in vulnerability between CA1 and CA3 pyramidal neurons to an ischemic insult. PMID- 16045485 TI - The acute and the long-term effects of nigral lipopolysaccharide administration on dopaminergic dysfunction and glial cell activation. AB - Sustained reactive microgliosis may contribute to the progressive degeneration of nigral dopaminergic neurons in Parkinson's disease (PD), in 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine (MPTP) exposed human and in non-human primates. However, the temporal relationship between glial cell activation and nigral cell death is relatively unexplored. Consequently, the effects of acute (24 h) and chronic (30 days) glial cell activation induced by unilateral supranigral lipopolysaccharide (LPS) administration were studied in rats. At 24 h, LPS administration caused a marked reduction in the number of tyrosine hydroxylase immunoreactive (TH-ir) neurons in the substantia nigra (SN) but striatal TH-ir was unaffected. By 30 days, the loss of TH-positive neurons in the LPS-treated nigra was no greater than at 24 h although a heterogeneous loss of striatal TH-ir was present. The loss of nigrostriatal neurons was of functional significance, as at 30 days, LPS-treated rats exhibited ipsiversive circling in response to (+) amphetamine administration. At 24 h, there was a moderate increase in glial fibrillary acidic protein (GFAP)-ir astrocytes in the SN but a marked elevation of p47phox positive OX-42-ir microglia, and intense inducible nitric oxide synthase (iNOS)-ir and 3-nitrotyrosine (3-NT)-ir was present. However, by 30 days the morphology of OX-42-ir microglia returned to a resting state, the numbers were greatly reduced and no 3-NT-ir was present. At 30 days, GFAP-ir astrocytes were markedly increased in number and iNOS-ir was present in fibrillar astrocyte like cells. This study shows that acute glial activation leading to dopaminergic neuron degeneration is an acute short-lasting response that does not itself perpetuate cell death or lead to prolonged microglial activation. PMID- 16045486 TI - Birth-date-dependent segregation of the mouse cerebral cortical neurons in reaggregation cultures. AB - Cerebral cortical neurons form a six-layered structure in which their position depends on their birth date. This developmental process requires the presence of Reelin, which is secreted by Cajal-Retzius cells in the cortical marginal zone (MZ). However, it is still unclear whether the migration from the ventricular zone (VZ) to beneath the MZ is essential for the neurons to segregate into layers. Previous transplantation studies of ferret cerebral cortical neurons suggested that their ultimate laminar fate is, at least to some extent, determined in the VZ but it is unknown how 'laminar fate' eventually positions cells in a specific layer. To explore the segregation properties of mouse cortical cells that have not yet arrived beneath the MZ, embryonic day (E)16 VZ and intermediate zone (IMZ) cells were dissociated and allowed to reaggregate for 1-4 days in vitro. The results suggested that the migrating neurons in the IMZ at E16 preferentially located near the centre of the aggregates, more than did the proliferative cells from the VZ. The birth-date labelling followed by the dissociation-reaggregation culture suggested that the segregation properties of the E16 IMZ was characteristic of the E14-born cells, which were migrating in the IMZ at E16, but they were not general properties of migrating IMZ cells. This birth-date-dependent segregation mechanism was also observed in the Reelin signalling-deficient yotari cells. These findings suggest that cortical neurons acquire a birth-date-dependent segregation mechanism before their somas reach the MZ. PMID- 16045487 TI - Differentiation of an auditory neuronal cell line suitable for cell transplantation. AB - The auditory neuroblast cell line US/VOT-N33 (N33), which is conditionally immortal, was studied as an in vitro model for the differentiation of spiral ganglion neurons (SGNs) and as a candidate for cell transplantation in rodents. It expresses numerous molecular markers characteristic of auditory neuroblasts, including the transcription factors GATA3, NeuroD, Brn3a and Islet1, as well as the neuronal cytoskeletal protein beta3-tubulin. It displays active migratory behaviour in vitro and in vivo. In the presence of the fibroblast growth factors FGF1 or FGF2 it differentiates bipolar morphologies similar to those of native SGNs. In coculture with neonatal cochlear tissue it is repelled from epithelial surfaces but not from native SGNs, alongside which it extends parallel neuronal processes. When injected into the retina in vivo, EGFP-labelled N33 cells were traced for 1-2 weeks and migrated rapidly within the subretinal space. Cells that found their way into the retinal ganglion cell layer extended multiple processes but did not express beta3-tubulin. The ability of N33 to migrate, to differentiate, to localize with native SGNs in vitro and to survive in vivo suggests that they provide an effective model for SGN differentiation and for cell transplantation into the ear. PMID- 16045488 TI - Nociceptive spinal withdrawal reflexes but not spinal dorsal horn wide-dynamic range neuron activities are specifically inhibited by halothane anaesthesia in spinalized rats. AB - The aim of the present study was to investigate the spinal cord effects and sites of action of different inhaled concentrations (0.5-2%) of the anaesthetic, halothane. Simultaneous recordings were made of 3 Hz, suprathreshold (1.5 x T) electrically evoked spinal dorsal horn (DH) wide-dynamic range (WDR) neuron responses and of single motor unit (SMU) electromyographic (EMG) responses underlying the spinal withdrawal reflex in spinalized Wistar rats. Compared with the baseline responses obtained with 0.5% halothane, the electrically evoked early responses of the DH WDR neurons as well as the SMUs were only depressed by the highest, 2% concentration of halothane. In contrast, 1.5% halothane markedly inhibited the late responses of the DH WDR neurons, whereas 1% halothane started to significantly depress the late responses of the SMUs. Likewise, wind-up of the WDR neuron late responses was inhibited by 1.5-2% halothane, whereas 1-2% halothane significantly depressed wind-up of the SMU EMG late responses. The inhibitory effects of 2% halothane on the early and the late responses of the DH WDR neurons, but not of the SMUs, were completely reversed by opioid micro receptor antagonist naloxone (0.04 mg/kg). However, no significant effects of naloxone were found on different responses of the DH WDR neurons as well as the SMUs at 0.5-1% halothane, suggesting that different concentrations of halothane may modulate different spinal receptors. We conclude that halothane at high concentrations (1.5-2%) seems to play a predominant inhibitory role via spinal multireceptors on ventral horn (VH) motor neurons, and less on DH sensory WDR neurons, of the spinal cord. PMID- 16045489 TI - Inhibition of peripheral vanilloid TRPV1 receptors reduces noxious heat-evoked responses of dorsal horn neurons in naive, carrageenan-inflamed and neuropathic rats. AB - The vanilloid TRPV1 receptor, present on primary afferent fibres, is activated by noxious heat, low pH and endogenous vanilloids. Changes in the function or distribution of TRPV1 receptors may play an important role in pain induced by inflammation or neuropathy. The aim of the present study was to evaluate the role of peripheral TRPV1 receptors in thermal nociception in rat models of inflammatory and neuropathic pain. Here, we have determined the effects of peripheral administration of the potent TRPV1 receptor antagonist iodoresiniferatoxin (IRTX) on noxious heat (45 degrees C)-evoked responses of spinal wide dynamic range (WDR) neurons in naive, carrageenan-inflamed, sham operated and L5/6 spinal nerve-ligated (SNL) anaesthetized rats in vivo. In addition, effects of peripheral administration of IRTX on mechanically evoked responses of WDR neurons were determined in sham-operated and SNL rats. Carrageenan inflammation significantly (P<0.05) increased the 45 degrees C-evoked responses of WDR neurons. Intraplantar injection of the lower dose of IRTX (0.004 microg) inhibited (P<0.05) 45 degrees C-evoked responses of WDR neurons in carrageenan-inflamed, but not in naive, rats. The higher dose of IRTX (0.4 microg) significantly (P<0.05) inhibited 45 degrees C-evoked responses in both inflamed and naive rats. In sham-operated and SNL rats, IRTX (0.004 and 0.4 microg) significantly (P<0.05) inhibited 45 degrees C-evoked, but had no effect on mechanically evoked responses of WDR neurons. These data support the role of peripheral TRPV1 receptors in noxious thermal transmission in naive, inflamed and neuropathic rats, and suggest that there is an increased functional contribution of peripheral TRPV1 receptors following acute inflammation. PMID- 16045490 TI - Inhibitory effects of CB1 and CB2 receptor agonists on responses of DRG neurons and dorsal horn neurons in neuropathic rats. AB - Cannabinoid 2 (CB2) receptor mediated antinociception and increased levels of spinal CB2 receptor mRNA are reported in neuropathic Sprague-Dawley rats. The aim of this study was to provide functional evidence for a role of peripheral, vs. spinal, CB2 and cannabinoid 1 (CB1) receptors in neuropathic rats. Effects of the CB2 receptor agonist, JWH-133, and the CB1 receptor agonist, arachidonyl-2 chloroethylamide (ACEA), on primary afferent fibres were determined by calcium imaging studies of adult dorsal root ganglion (DRG) neurons taken from neuropathic and sham-operated rats. Capsaicin (100 nm) increased [Ca2+]i in DRG neurons from sham and neuropathic rats. JWH-133 (3 microm) or ACEA (1 microm) significantly (P<0.001) attenuated capsaicin-evoked calcium responses in DRG neurons in neuropathic and sham-operated rats. The CB2 receptor antagonist, SR144528, (1 microm) significantly inhibited the effects of JWH-133. Effects of ACEA were significantly inhibited by the CB1 receptor antagonist SR141716A (1 microm). In vivo experiments evaluated the effects of spinal administration of JWH-133 (8-486 ng/50 microL) and ACEA (0.005-500 ng/50 microL) on mechanically evoked responses of neuropathic and sham-operated rats. Spinal JWH-133 attenuated mechanically evoked responses of spinal neurons in neuropathic, but not sham operated rats. These inhibitory effects were blocked by SR144528 (0.001 microg/50 microL). Spinal ACEA inhibited mechanically evoked responses of neuropathic and sham-operated rats, these effects were blocked by SR141716A (0.01 microg/50 microL). Our data provide evidence for a functional role of CB2, as well as CB1 receptors on DRG neurons in sham and neuropathic rats. At the level of the spinal cord, CB2 receptors have inhibitory effects in neuropathic, but not sham-operated rats suggesting that spinal CB2 may be an important analgesic target. PMID- 16045491 TI - Gestational nicotine exposure reduces nicotinic cholinergic receptor (nAChR) expression in dopaminergic brain regions of adolescent rats. AB - Children of women who smoked during pregnancy are at increased risk of dependence when smoking is initiated during adolescence. We previously reported that gestational nicotine exposure attenuated dopamine release induced by nicotine delivered during adolescence. In this study, we determined the effects of gestational nicotine exposure on nicotinic cholinergic receptor (nAChR) expression. Timed pregnant rats received nicotine (2 mg/kg/day) or vehicle via mini-osmotic pumps during gestation. Treatments continued in pups via maternal nursing during postnatal days (PN) 2-14 (equivalent to the human in utero third trimester). On PN35, 125I-epibatidine binding to nAChR was measured. The Bmax values (fmol/mg) in prefrontal cortex (PFC), nucleus accumbens (NAcc), substantia nigra (SN) and ventral tegmental area (VTA) were reduced by 26.6% (P<0.05), 32.6% (P<0.01), 23.0% (P<0.01) and 27.6% (P<0.05), respectively. In addition, gender differences were found in vehicle-treated groups; in SN and VTA, females were 79.3% (P<0.005) and 82.9% (P=0.08) of males, respectively. The expression of nAChR subunit mRNAs was measured using real-time RT-PCR on laser-capture microdissected tissues. In adolescent VTA, gestational nicotine exposure reduced (P<0.05) nAChR subunit mRNAs encoding alpha3 (53.0%), alpha4 (23.9%), alpha5 (46.7%) and beta4 (61.4%). In NAcc core, the treatment increased alpha3 mRNA (75.8%). In addition, the number of neurons in VTA was reduced by 15.0% (P<0.001). These studies indicate that gestational exposure to nicotine induces long-lasting changes in nAChR expression that may underlie the vulnerability of adolescents to dependence on nicotine. PMID- 16045492 TI - Sonic hedgehog signalling in neurons of adult ventrolateral nucleus tractus solitarius. AB - The transmembrane receptor Patched (Ptc) mediates the action of the diffusing factor Sonic hedgehog (Shh), which is implicated in establishing morphogenetic gradients during embryonic development. Whereas alteration of Ptc function is associated with developmental abnormalities and brain tumors, its functional activity and roles in the adult brain have yet to be elucidated. Here we describe the complementary pattern of Shh and Ptc expression in the rat dorsal vagal motor nucleus and the ventrolateral nucleus tractus solitarius (vNTS), respectively. Those two interconnected structures regulate the cardiorespiratory function during hypoxia. Bath application of a subnanomolar concentration of aminoterminal Shh protein (ShhN) to a slice preparation of the vNTS induces a rapid decrease in neuronal firing followed by a bursting activity that propagates in the neuronal network. Intracellular current injections show that bursts result from an action on the neuronal membrane electro-responsiveness. Both inhibiting and bursting effects are blocked by the monoclonal Shh antibody 5E1 and may require the Ptc binding site of ShhN. Thus, ShhN acting on specific neuronal sites controls electrophysiological properties of differentiated neurons of the vNTS. We speculate on a retrocontrol of cardiorespiratory signals in the vNTS, by Shh generated in dorsal vagal motoneurons. PMID- 16045493 TI - Activity-dependent subcellular localization of NAC1. AB - The expression of the transcriptional regulator NAC1 is increased in the nucleus accumbens of rats withdrawn from cocaine self-administration, and in vivo studies indicate that the up-regulation is a compensatory mechanism opposing the acute effects of cocaine. Both mammalian two-hybrid assay and punctate localization largely in the nucleus suggest NAC1 is a transcriptional regulator. However, in this report it is shown that in differentiated PC12 and Neuro2A cells, as well as in primary cortical neurons, NAC1 is diffusely expressed not only in the cell nucleus but also in cytoplasm. Blockade of spontaneous electrical activity by tetrodotoxin prevented the diffuse expression of NAC1, and depolarization with high potassium concentrations induced diffuse cellular localization in non differentiating cells. The use of protein kinase C (PKC) inhibitors and activator, as well as the systematic mutation of potential PKC phosphorylation sites in NAC1, demonstrated that phosphorylation of residue S245 by PKC is a necessary event inducing diffuse NAC1 expression outside of the nucleus. These observations indicate a potential non-transcriptional role for NAC1 in the brain. PMID- 16045494 TI - Expression of zonula occludens-1 (ZO-1) and the transcription factor ZO-1 associated nucleic acid-binding protein (ZONAB)-MsY3 in glial cells and colocalization at oligodendrocyte and astrocyte gap junctions in mouse brain. AB - The PDZ domain-containing protein zonula occludens-1 (ZO-1) interacts with several members of the connexin (Cx) family of gap junction-forming proteins and has been localized to gap junctions, including those containing Cx47 in oligodendrocytes. We now provide evidence for ZO-1 expression in astrocytes in vivo and association with astrocytic connexins by confocal immunofluorescence demonstration of ZO-1 colocalization with astrocytic Cx30 and Cx43, and by ZO-1 coimmunoprecipitation with Cx30 and Cx43. Evidence for direct interaction of Cx30 with ZO-1 was obtained by pull-down assays that indicated binding of Cx30 to the second of the three PDZ domains in ZO-1. Further, we investigated mouse Y-box transcription factor MsY3, the canine ortholog of which has been termed ZO-1 associated nucleic acid-binding protein (ZONAB) and previously reported to interact with ZO-1. By immunofluorescence using specific antimouse ZONAB antibody, ZONAB was found to be associated with oligodendrocytes throughout mouse brain and spinal cord, and to be colocalized with oligodendrocytic Cx47 and Cx32 as well as with astrocytic Cx43. Our results extend the CNS cell types that express the multifunctional protein ZO-1, demonstrate an additional connexin (Cx30) that directly interacts with ZO-1, and show for the first time the association of a transcription factor (ZONAB) with ZO-1 localized to oligodendrocyte and astrocyte gap junctions. Given previous observations that ZONAB and ZO-1 in combination regulate gene expression, our results suggest roles of glial gap junction-mediated anchoring of signalling molecules in a wide variety of glial homeostatic processes. PMID- 16045495 TI - Multisensory responses and receptive field properties of neurons in the substantia nigra and in the caudate nucleus. AB - The basal ganglia are widely regarded as structures involved in sensorimotor co ordination, but little is known about the sensory background of their function. We publish here descriptions of the excitatory sensory responses and receptive field properties of the visual, auditory, somatosensory and multisensory caudate nucleus and substantia nigra pars reticulata neurons. Altogether 111 caudate nucleus and 124 substantia nigra sensory neurons were recorded in halothane anaesthetized, immobilized, artificially ventilated cats. The sensory properties of the caudate and nigral neurons were found to be quite similar. A majority of the units were unimodal while a significant proportion of them were multisensory. The visual and the somatosensory modalities predominated for both nuclei. The sensory receptive fields were extremely large. The visual and auditory receptive fields covered the whole physically approachable sensory field and the somatosensory receptive fields covered the whole body surface of the animal. The receptive field properties of the multisensory caudate and nigral units were similar to those of the unimodal neurons. We observed no signs of retinotopical or somatotopical organization within the basal ganglia. The particular sensory receptive field properties, together with the significant number of multisensory neurons in the basal ganglia, suggest the existence of a multisensory pathway of presumably tectal origin involving the caudate nucleus and the substantia nigra that may serve for the sensory feedback of motor actions co-ordinated by the basal ganglia. PMID- 16045496 TI - Increased measures of anxiety and weight gain in mice lacking the group III metabotropic glutamate receptor mGluR8. AB - To study the role of the metabotropic glutamate receptor 8 (mGluR8), mice lacking this receptor were generated by homologous recombination. Homozygous mGluR8 deficient mice are about 8% heavier than their wild-type age-matched controls after reaching 4 weeks of age. This weight difference is not caused by an altered food intake and is not exacerbated by feeding the animals a high-fat diet. Moreover, mGluR8-/- mice are mildly insulin resistant, possibly as a result of being overweight. Behavioral testing revealed a reduced locomotor activity of mGluR8-/- mice compared with wild-type mice during the first 3 days in a novel enclosed environment. However after 3 days, the locomotor activities of wild-type and mGluR8-/- mice were similar, suggesting a reduced exploratory behavior of mGluR8-/- mice in a novel enclosed environment. By contrast, there were no genotype differences in locomotor activity in the open field, plus maze, or in total time spent exploring objects during object recognition tests, indicating that there is a dissociation between effects of mGluR8 deficiency in exploratory activity in a novel safe enclosed environment vs. a more anxiogenic novel open environment. The absence of mGluR8 also leads to increased measures of anxiety in the open field and elevated plus maze. Whether the diverse phenotypic differences observed in mGluR8-/- mice result from the misregulation of a unique neural pathway, possibly in the thalamus or hypothalamus, or whether they are the consequence of multiple developmental and functional alterations in synaptic transmission, remains to be determined. PMID- 16045497 TI - S-cones do not contribute to the OFF-midget pathway in the retina of the marmoset, Callithrix jacchus. AB - It is well established that in primate retina both medium- and long-wavelength sensitive cone types provide input to the midget-parvocellular pathway. The question, however, whether short-wavelength-sensitive (S or 'blue') cones provide input to the OFF-division of the midget-parvocellular pathway is still controversial. In the present study, we investigated the connections of nearly 400 S-cones with OFF-midget bipolar cells in central and peripheral retina of a New World monkey, the marmoset. Horizontal sections or pieces of whole retinae were double-labelled with an antiserum to S-cone opsin to identify S-cones and antibodies to the cell adhesion molecule CD15 to identify OFF-midget bipolar cells. Peanut agglutinin coupled to a fluorescent tag was used to label the cone pedicles of all cone types. Peanut agglutinin was also used to distinguish S cones from the other cone types. The sections were analysed with deconvolution microscopy. We found that nearly all pedicles of medium- and long-wavelength sensitive cones are located opposite distinct dendritic clusters formed by OFF midget bipolar cells. By contrast, the S-cone pedicles are not located opposite dendritic clusters. Instead, S-cones make sparse contacts with CD15-labelled processes. Some of these processes protruded from OFF-midget bipolar clusters, whereas others could be traced to a diffuse bipolar cell type. Thus, in the marmoset retina the midget-parvocellular system does not carry a blue-OFF signal. PMID- 16045498 TI - Activity of substantia nigra pars reticulata neurons during smooth pursuit eye movements in monkeys. AB - Nuclei within the basal ganglia (BG), in particular the substantia nigra pars reticulata (SNr), subthalamic nucleus (STN) and caudate nucleus, are known to be involved in the generation of rapid or saccadic eye movements. Neurons in the SNr are active tonically and generally show a pause, but also increase, in discharge rate, for the appearance of visual stimuli and the generation of saccades. Recent experimental results in oculomotor regions of the brainstem reveal overlap in the neuronal pathways used for saccades and smooth pursuit, or slow tracking, eye movements. Whether the overlap of processing for saccades and pursuit extends to the oculomotor BG is unknown. In the present report, we were interested in whether the overlap between the pursuit and saccadic systems extends into the oculomotor BG. Using single-neuron recording and electrical stimulation techniques, we tested whether neurons within the saccade portion of the BG, the SNr, could be involved in smooth pursuit eye movements. Monkeys were required to follow visual targets with either a smooth eye movement or a saccade while we recorded from SNr neurons. We report here on SNr neuronal activity that was modulated during the performance of visually guided saccades and also during the initiation and the maintenance of smooth pursuit eye movements. Importantly, the modulation of neuronal activity during pursuit was present even when catch-up saccades were absent. The majority of SNr neurons was active tonically and their discharge ceased during pursuit, although some neurons also increased their discharge rate during smooth pursuit, similar to the behaviour reported for saccades. We also found that electrical stimulation of the SNr during the initiation of pursuit suppressed ipsiversive and, in some cases, enhanced contraversive pursuit. Our combined recording and stimulation results are consistent with the hypothesis that the overlap between the pursuit and saccadic systems extends, at least somewhat, into the BG and that the signal conveyed by the SNr can be used by the pursuit system. Like the signal for saccades, the SNr may provide a permissive disinhibition for pursuit eye movements. We hypothesize that alterations in this signal in BG diseased states such as Parkinson's may explain in part the deficits observed in smooth pursuit eye movements of these patients. PMID- 16045499 TI - Neural responses in cat visual cortex reflect state changes in correlated activity. AB - Cortical state is characterized by ongoing rhythmic neural activity. Changes in rhythmic activity and thus in cortical state are shown to occur spontaneously in the anesthetized cat. We were interested in whether these state changes have an affect on the cortical processing of sensory stimuli. This was investigated by recording spontaneous and stimulus-evoked local field potentials and multi-unit neuronal activity (MUA) from trans-cortical electrode arrays in the visual cortex of the anesthetized cat. Changes in cortical state were identified by calculating the cross-correlation strength and cross-coherency, between MUA channels at different layers and on separate electrode arrays. Spontaneous changes in rhythmic activity were associated with changes in the strength of stimulus-evoked multiple unit responses of cortical neurons. The highest multi-unit responses were found in periods when low-frequency rhythms of the electroencephalogram increase in magnitude and high-frequency rhythms decrease. Such changes in evoked responses were maximal at layer IV, the input layer of the visual cortex. Our findings suggest that stimulus response magnitude depends on rhythmic state and reflects changes in functional connectivity within the visual cortex. PMID- 16045500 TI - Frequency-dependent impairment of hippocampal LTP from NMDA receptors with reduced calcium permeability. AB - Changes in postsynaptic Ca2+ levels are essential for alterations in synaptic strength. At hippocampal CA3-to-CA1 synapses, the Ca2+ elevations required for LTP induction are typically mediated by NMDA receptor (NMDAR) channels but a contribution of NMDAR-independent Ca2+ sources has been implicated. Here, we tested the sensitivity of different protocols modifying synaptic strength to reduced NMDAR-mediated Ca2+ influx by employing mice genetically programmed to express in forebrain principal neurons an NR1 form that curtails Ca2+ permeability. Reduced NMDAR-mediated Ca2+ influx did not facilitate synaptic depression in CA1 neurons of these genetically modified mice. However, we observed that LTP could not be induced by pairing low frequency synaptic stimulation (LFS pairing) with postsynaptic depolarization, a protocol that induced robust LTP in wild-type mice. By contrast to LFS pairing, similar LTP levels were generated in both genotypes when postsynaptic depolarization was paired with high frequency synaptic stimulation (HFS). This indicates that the postsynaptic Ca2+ elevation also reached threshold during HFS in the mutant, probably due to summation of NMDAR-mediated Ca2+ influx. However, only in wild type mice did repeated HFS further enhance LTP. All tested forms of LTP were blocked by the NMDAR antagonist D-AP5. Collectively, our results indicate that only NMDAR-dependent Ca2+ sources (NMDARs and Ca2+-dependent Ca2+ release from intracellular stores) mediate LFS pairing-evoked LTP. Moreover, LTP induced by the first HFS stimulus train required lower Ca2+ levels than the additional LTP obtained by repeated trains. PMID- 16045501 TI - Roles of NMDA receptor NR2A and NR2B subtypes for long-term depression in the anterior cingulate cortex. AB - The anterior cingulate cortex (ACC) is thought to be important for the establishment, consolidation and retrieval of permanent memory. In many brain regions, including the hippocampus, it is suggested that long-term potentiation (LTP) and long-term depression (LTD), the cellular mechanisms for learning and memory, require the activation of glutamate N-methyl-D-aspartate receptors (NMDARs). In the hippocampus, the NR2A subunit is believed to be involved in the induction of LTP, whereas the NR2B subunit contributes to the formation of LTD. However, LTD has been less well studied in the ACC as compared with the hippocampus and little is known about the role of NMDA subtype receptors in cingulate LTD. Here we show that LTD can be induced by the combination of presynaptic stimulation with postsynaptic depolarization ('pairing training') in adult mouse ACC neurons. This form of LTD is an NMDAR- and voltage-dependent mechanism and a postsynaptic Ca2+ increase is required for the induction of LTD. Furthermore, our studies provide direct physiological evidence that both NR2A and NR2B subunits are involved in the induction of LTD in the ACC. PMID- 16045502 TI - How does transcranial DC stimulation of the primary motor cortex alter regional neuronal activity in the human brain? AB - Transcranial direct current stimulation (tDCS) of the primary motor hand area (M1) can produce lasting polarity-specific effects on corticospinal excitability and motor learning in humans. In 16 healthy volunteers, O positron emission tomography (PET) of regional cerebral blood flow (rCBF) at rest and during finger movements was used to map lasting changes in regional synaptic activity following 10 min of tDCS (+/-1 mA). Bipolar tDCS was given through electrodes placed over the left M1 and right frontopolar cortex. Eight subjects received anodal or cathodal tDCS of the left M1, respectively. When compared to sham tDCS, anodal and cathodal tDCS induced widespread increases and decreases in rCBF in cortical and subcortical areas. These changes in rCBF were of the same magnitude as task related rCBF changes during finger movements and remained stable throughout the 50-min period of PET scanning. Relative increases in rCBF after real tDCS compared to sham tDCS were found in the left M1, right frontal pole, right primary sensorimotor cortex and posterior brain regions irrespective of polarity. With the exception of some posterior and ventral areas, anodal tDCS increased rCBF in many cortical and subcortical regions compared to cathodal tDCS. Only the left dorsal premotor cortex demonstrated an increase in movement related activity after cathodal tDCS, however, modest compared with the relatively strong movement independent effects of tDCS. Otherwise, movement related activity was unaffected by tDCS. Our results indicate that tDCS is an effective means of provoking sustained and widespread changes in regional neuronal activity. The extensive spatial and temporal effects of tDCS need to be taken into account when tDCS is used to modify brain function. PMID- 16045503 TI - Blockade of NMDA receptors in the dorsomedial striatum prevents action-outcome learning in instrumental conditioning. AB - Although there is consensus that instrumental conditioning depends on the encoding of action-outcome associations, it is not known where this learning process is localized in the brain. Recent research suggests that the posterior dorsomedial striatum (pDMS) may be the critical locus of these associations. We tested this hypothesis by examining the contribution of N-methyl-D-aspartate receptors (NMDARs) in the pDMS to action-outcome learning. Rats with bilateral cannulae in the pDMS were first trained to perform two actions (left and right lever presses), for sucrose solution. After the pre-training phase, they were given an infusion of the NMDA antagonist 2-amino-5-phosphonopentanoic acid (APV, 1 mg/mL) or artificial cerebral spinal fluid (ACSF) before a 30-min session in which pressing one lever delivered food pellets and pressing the other delivered fruit punch. Learning during this session was tested the next day by sating the animals on either the pellets or fruit punch before assessing their performance on the two levers in extinction. The ACSF group selectively reduced responding on the lever that, in training, had earned the now devalued outcome, whereas the APV group did not. Experiment 2 replicated the effect of APV during the critical training session but found no effect of APV given after acquisition and before test. Furthermore, Experiment 3 showed that the effect of APV on instrumental learning was restricted to the pDMS; infusion into the dorsolateral striatum did not prevent learning. These experiments provide the first direct evidence that, in instrumental conditioning, NMDARs in the dorsomedial striatum are involved in encoding action-outcome associations. PMID- 16045504 TI - The role of the dorsomedial striatum in instrumental conditioning. AB - Considerable evidence suggests that, in instrumental conditioning, rats learn the relationship between actions and their specific consequences or outcomes. The present study examined the role of the dorsomedial striatum (DMS) in this type of learning after excitotoxic lesions and reversible, muscimol-induced inactivation. In three experiments, rats were first trained to press two levers for distinct outcomes, and then tested after training using a variety of behavioural assays that have been established to detect action-outcome learning. In Experiment 1, pre-training lesions of the posterior DMS abolished the sensitivity of rats' instrumental performance to both outcome devaluation and contingency degradation when tested in extinction, whereas lesions of the anterior DMS had no effect. In Experiment 2, both pre-training and post-training lesions of the posterior DMS were equally effective in reducing the sensitivity of performance both to devaluation and degradation treatments. In Experiment 3, the infusion of muscimol into the posterior DMS selectively abolished sensitivity of performance to devaluation and contingency degradation without impairing the ability of rats to discriminate either the instrumental actions performed or the identity of the earned outcomes. Taken together, these results suggest that the posterior region of the DMS is a crucial neural substrate for the acquisition and expression of action-outcome associations in instrumental conditioning. PMID- 16045505 TI - The robustness of perception. AB - The natural environment around us, which is often crowded, cluttered or even foggy, is subject to a dynamically changing composition of objects and events. The human brain is continuously perceiving, recognizing and evaluating this dynamic scene composition. If the perception of degraded visual objects is important, e.g. in the case of potential threat stimuli, the brain needs to be more sensitive in detecting these objects from the natural environment. It is therefore hypothesized that reacting to the dynamically changing environment involves a robust and quick processing of salient information, which can be either with or without conscious awareness. We investigated the dynamics and robustness of perception using pictures of three salience levels, i.e. fearful faces (most salient), neutral faces (salient) and houses (nonsalient), which appear from dynamically decreasing random visual noise. Stimuli were matched for luminance, contrast, brightness and spatial frequency information. Reaction times show a significantly earlier response for faces than for houses. Fearful faces were significantly more quickly detected than neutral faces. The neural correlates sustaining robust perception were investigated with event-related functional magnetic resonance imaging (fMRI). The amygdala showed a significant perception-related response for faces, as compared to houses, that was further enhanced for fearful faces as compared to neutral faces. Our data indicate that emotionally salient information processing is (i) mediated by the amygdala and (ii) more robust than for nonsalient stimuli as it shows a significantly lower perceptual threshold. PMID- 16045506 TI - Hypocretin/orexin-containing neurons are produced in one sharp peak in the developing ventral diencephalon. AB - The birth date of hypocretin-containing neurons was analysed using the bromodeoxyuridine method in the rat. The results indicate that these neurons are generated between embryonic days 11 (E11) and E14, with a sharp peak on E12. This spatiotemporal pattern of genesis contrasts with that of the co-distributed neurons producing the melanin-concentrating hormone in the lateral hypothalamic area, which have been described as generated in one large peak from E10 to E16. These observations may be linked to the relative distribution area of both populations. PMID- 16045507 TI - The role of the human posterior parietal cortex in memory-guided saccade execution: a double-pulse transcranial magnetic stimulation study. AB - The present study investigated the role of the right posterior parietal cortex (PPC) in the triggering of memory-guided saccades by means of double-pulse transcranial magnetic stimulation (dTMS). Shortly before saccade onset, dTMS with different interstimulus intervals (ISI; 35, 50, 65 or 80 ms) was applied. For contralateral saccades, dTMS significantly decreased saccadic latency with an ISI of 80 ms and increased saccadic gain with an ISI of 65 and 80 ms. Together with the findings of a previous study during frontal eye field (FEF) stimulation the present results demonstrate similarities and differences between both regions in the execution of memory-guided saccades. Firstly, dTMS facilitates saccade triggering in both regions, but the timing is different. Secondly, dTMS over the PPC provokes a hypermetria of contralateral memory-guided saccades that was not observed during FEF stimulation. The results are discussed within the context of recent neurophysiological findings in monkeys. PMID- 16045510 TI - The benefits and risks of inducing labour in patients with prior caesarean delivery: a systematic review. AB - OBJECTIVE: To evaluate the risks and benefits of inducing labour in women with a prior caesarean delivery. DESIGN: Systematic review. SAMPLE: Pregnant women with prior caesarean delivery. METHODS: Studies were identified using MEDLINE, EMBASE, HealthSTAR, Cochrane Database of Systematic Reviews, Cochrane Controlled Trials Register, Database of Abstracts of Reviews of Effectiveness, reference lists and experts. MAIN OUTCOME MEASURES: All studies reporting data for outcomes in women with induced labours and prior caesarean were eligible. Methodologic quality was evaluated using the criteria of the U.S. Preventive Services Task Force and the NHS Centre for Reviews and Dissemination. We assigned studies good, fair or poor rating. RESULTS: We reviewed 162 full text articles, identified 14 fair-quality studies, and found no good-quality studies. Compared with spontaneous labour, induction was more likely to result in caesarean delivery. Of women undergoing spontaneous labour, 20% had a caesarean (range 11-35%) compared with 32% receiving oxytocin (range 18-44%). In studies of PGE2, spontaneous labour resulted in caesarean delivery in 24% (range 18-51%) compared with 48% with PGE2 (range 28-51%). There was a non-significant increase in uterine ruptures among those induced compared with spontaneous labours. There were no maternal deaths; other maternal complications were infrequently reported. Only four studies reported on infant deaths; other infant outcomes were inadequately reported. CONCLUSION: Women with a history of caesarean attempting trial of labour who require induction have a higher rate of caesarean delivery and have a slightly elevated risk of rupture compared with similar women with spontaneous labour. More consideration is needed for potential confounders: dose, reasons for induction and appropriate comparison groups. PMID- 16045511 TI - Non-invasive intrapartum fetal ECG: preliminary report. AB - OBJECTIVES: To obtain fetal heart rate, detailed fetal electrocardiography (fECG) signals and uterine contractions during labour using a single device. DESIGN: Prospective observational study. SETTING: Delivery suite at a tertiary referral hospital, London, UK. POPULATION: Fifteen patients at median gestation of 39 weeks (range 24-41) were recruited at median cervical dilatation of 4.0 cm (range 0-10) of whom 8/15 (53%) had intact amniotic membranes. METHODS: Using 12 abdominally sited electrodes, we recorded the composite abdominal signal in pregnancies intrapartum. The recorded data were analysed off-line using a blind signal separation technique. MAIN OUTCOME MEASURES: Success of signal separation and fECG time intervals. RESULTS: Successful fECG signal acquisition was achieved in 12/15 (80%) patients and an averaged fECG waveform acquired. In these patients, P and QRS waves were seen in all cases, and T waves in 11/12 (92%). True beat-to-beat heart rate (HR) was displayed and measures of its variability obtained. The mother's ECG and uterine electrical activity, shown to match tocographically recorded uterine contractions, were also separated and displayed. Failure to acquire fECG in three cases was attributed to excessive abdominal muscular activity and electrical interference. CONCLUSIONS: This study demonstrates a non-invasive technique that displays detailed intrapartum fECG waveforms, HR variability, maternal ECG and uterine contractions simultaneously, all in a single device and which avoids the potential risks of invasive monitoring with a fetal scalp electrode. PMID- 16045512 TI - Recreational drugs and fetal gastroschisis: maternal hair analysis in the peri conceptional period and during pregnancy. AB - OBJECTIVE: To objectively measure individual recreational drug groups in maternal hair samples timed for the period of conception and different stages of pregnancy in expectant mothers with a diagnosis of fetal gastroschisis. DESIGN: Prospective observational study. SETTING: Tertiary level Fetal Medicine Unit in a University teaching hospital. POPULATION: Pregnant women with a diagnosis of fetal gastroschisis (n= 22) and a control group of women (n= 25) with a normal fetus. METHODS: Hair samples were cut from the vertex of the head of expectant mothers with a diagnosis of fetal gastroschisis and a matched control group in whom the fetus was normal. The samples were analysed by enzyme-linked immunosorbent assay (ELISA) for individual drug groups. Confirmatory tests using gas chromatography with mass spectrometry (GCMS) were used. MAIN OUTCOME MEASURES: Presence of recreational drug compounds in hair samples. RESULTS: In the group with fetal gastroschisis there were four proven positive cases for recreational drug abuse (18%) and there were none in the control group. CONCLUSIONS: The incidence of recreational drug use in the peri-conceptional period and the first trimester by expectant mothers with a diagnosis of fetal gastroschisis is 18%. This association may be linked to the recent increase in the incidence of gastroschisis among younger mothers. PMID- 16045513 TI - Customized birthweight centiles predict SGA pregnancies with perinatal morbidity. AB - OBJECTIVES: To determine the following: (1) the proportion of babies reclassified as small or appropriately grown using customized and population centiles; and (2) the relative risks of perinatal morbidity, including abnormal umbilical Doppler studies, in babies classified as small for gestational age (SGA) and appropriate for gestational age (non-SGA) using the two centile calculations. DESIGN: Cohort study in SGA and general hospital populations. SETTING: National Women's Hospital, Auckland, NZ. POPULATION: A cohort of SGA pregnancies (n= 374) and a general obstetric population (n= 12,879). METHODS: Pregnancy outcomes were compared between 'non-SGA both' (> or =10th% by population and customized centiles) and those who were 'SGA both' (<10th% by population and customized centiles), 'SGA customized only' (SGA by customized but non-SGA by population centiles) and 'SGA population only' (SGA by population but non-SGA by customized centiles). MAIN OUTCOME MEASURES: Maternal and newborn morbidity and perinatal death. RESULTS: In the SGA cohort 271 (72%) babies were 'SGA both', 27 (7%) were 'SGA customized only', 32 (9%) were 'population SGA only' and 44 (12%) were 'non SGA both'. In the general obstetric population 863 (6.7%) babies were 'SGA both', 445 (3.5%) were 'customized SGA only', 285 (2.2%) were 'population SGA only' and 11,286 (88%) were 'non-SGA both'. Perinatal death and newborn morbidity including nursery admission and long hospital stay were increased and comparable between 'SGA both' and 'customized SGA only' in both study populations. Newborn morbidity was low and comparable between 'population SGA only' and 'non-SGA both'. No perinatal deaths occurred in 'population SGA only' babies. Abnormal Doppler studies were more common in 'SGA both' or 'customized SGA only' but not in 'population SGA only' groups compared with 'non-SGA both'. CONCLUSIONS: Customized birthweight centiles identified small babies at risk of morbidity and mortality. Use of customized centiles is likely to detect more babies at risk of perinatal morbidity and mortality than would be detected by population centiles. PMID- 16045514 TI - Specific PGF(2alpha) receptor (FP) antagonism and human uterine contractility in vitro. AB - OBJECTIVE: PGF(2alpha) acts through its receptor, FP, as an important smooth muscle contractile agent. The aim of this study was to investigate the effects of specific FP antagonism, using the novel-specific FP non-competitive antagonist THG113.31, on spontaneous and agonist-elicited contractions in pregnant and non pregnant human myometrium in vitro. DESIGN: Scientific study. SETTING: University hospital and laboratories. Population Women undergoing caesarean section or hysterectomy. METHODS: Biopsies of human myometrium were obtained at elective caesarean section (n= 22) and from hysterectomy specimens from premenopausal women (n= 8). Dissected strips were mounted in tissue baths under physiological conditions. The effects of THG113.31 on spontaneous and oxytocin-induced contractions, in pregnant myometrium, and on phenylephrine-induced contractions, in non-pregnant myometrium, were measured. The effects of PGF(2alpha) on spontaneous contractions, in pregnant tissue, in the presence and absence of THG113.31, were investigated. The integrals of contractile activity measured were compared with those from simultaneously run control experiments. The pD(2) and mean maximal effect observed for THG113.31, and for PGF(2alpha) in the presence and absence of THG113.31, were calculated. MAIN OUTCOME MEASURES: Changes in contractility. RESULTS: THG113.31 exerted a potent relaxant effect in both spontaneous and oxytocin-induced contractility in pregnant tissue (P < 0.001), and phenylephrine-induced contractility in non-pregnant tissue (P < 0.001), compared with control experiments. PGF(2alpha) exerted a significant contractile effect on spontaneous contractions in pregnant tissue and this effect was not significantly attenuated by THG113.31 (P > 0.05). CONCLUSION: THG113.31 exerted a significant relaxant effect on human spontaneous and oxytocin-induced contractility but did not alter PGF(2alpha)-elicited contractility. These data raise questions about the exact mechanism of effect of THG113.31 and its interaction with FP. The uterorelaxant potency of THG113.31 in human myometrium in vitro indicates that it may be of limited use as a tocolytic compound. PMID- 16045515 TI - Obstetric management of a woman's first delivery and the implications for pelvic floor surgery in later life. AB - OBJECTIVE: To determine the influence of intrapartum care during a first delivery on the risk of pelvic floor surgery in later life. DESIGN: Nested case-control study with record linkage of a historical cohort and a current morbidity database. SETTING: Hospital births in Dundee 1952-1966. POPULATION: The 7556 primiparous women from the Walker cohort. METHODS: The cases (n= 352) were women who delivered a first singleton baby at term (> or =37 weeks) and subsequently had pelvic floor surgery. Controls (n= 1403) were women who delivered their first baby during the same time period and did not undergo surgery. Univariate and multivariate logistic regression analyses were performed taking account of demographic, anthropometric and obstetric factors. MAIN OUTCOME MEASURE: Pelvic floor surgery. RESULTS: Caesarean section was associated with a reduced risk of pelvic floor surgery compared with spontaneous vaginal delivery (odds ratio 0.16, 95% CI 0.05-0.55). Forceps delivery and infant birthweight >4.0 kg were not identified as significant risk factors (OR 0.94, 95% CI 0.71, 1.25, and OR 0.94, 95% CI 0.50, 1.75, respectively). Episiotomy and prolonged labour (>12 hours) may be associated risk factors but were of borderline significance (OR 1.46, 95% CI 0.99, 2.10, and OR 1.51, 95% CI 1.00, 2.27). CONCLUSION: Caesarean section in a first pregnancy appears to protect against pelvic floor surgery in later life. PMID- 16045516 TI - Breastfeeding expectations versus reality: a cluster randomised controlled trial. AB - OBJECTIVE: To evaluate the affect of an antenatal educational breastfeeding intervention on women's breastfeeding duration. DESIGN: Cluster randomised controlled trial. Unit of randomisation: electoral ward. The primary outcome was the proportion that fulfilled their antenatal breastfeeding expectation. Secondary outcomes were the number of women breastfeeding on discharge and at four months. Data were collected using a series of questionnaires and diaries. SETTING: Teaching hospital in North West of England. PARTICIPANTS: Women who expressed a desire to breast-feed at the start of their pregnancy. METHODS: Women were allocated to either routine antenatal education or an additional single educational group session supervised by a lactation specialist and attended by midwives from their locality. MAIN OUTCOME MEASURE: The proportion of women who fulfilled their expectation of breastfeeding. RESULT: One thousand three hundred and twelve women were randomised, with 1249 (95%) women available for analysis. There was no difference between the groups in the proportion of women who attained their expected duration of breastfeeding (OR 1.2; 95% CI 0.89-1.6; chi(2)= 1.4, df= 1, P= 0.2; mean cluster size 156, design effect 1.6). There were no differences between the groups in the uptake of breastfeeding on discharge (OR = 1.2; 95% CI 0.8-1.7; chi(2)= 1.1, df= 1, P= 0.3; mean cluster size 163, design effect = 2.0) or exclusively at four months (OR = 1.1; 95% CI 0.6-1.8; chi(2)= 0.07, df= 1, P= 0.8; mean cluster size 156, design effect 1.6). CONCLUSION: The provision of a single educational group session supervised by a lactation specialist, and attended by midwives and women, failed to promote the uptake of breastfeeding. Public health interventions, which encourage positive attitudes to breastfeeding within the family and wider community, should be developed and evaluated. PMID- 16045517 TI - Factors influencing repeat caesarean section: qualitative exploratory study of obstetricians' and midwives' accounts. AB - OBJECTIVE: To explore the views of health professionals on the factors influencing repeat caesarean section. DESIGN: Qualitative study involving semi structured interviews with professionals who care for women in pregnancy and labour. SETTING: Acute hospital trust with two maternity units and community midwifery service, Leicestershire, UK. SAMPLE: Twenty-five midwives and doctors. METHODS: Interviews with professionals were undertaken using a prompt guide. All interviews were audiotaped and transcribed verbatim. Analysis was based on the constant comparative method, assisted by QSR N5 software. MAIN OUTCOME MEASURES: Identification of factors influencing professional decision making about repeat caesarean section. RESULTS: Decision making in relation to repeat caesarean is a complex process involving several parties. Professionals identify the relevance of evidence for decision making for repeat caesarean. However, professionals feel that following strict protocols is of limited value because of the perceived substandard quality of evidence in this area, other external pressures and the contingent, unique and often unanticipated features of each case. Professionals also perceive that the organisation of care plays an important role in rates of repeat caesarean. CONCLUSIONS: Decision making for repeat caesarean is a social practice where standardised protocols may have limited value. Attention needs to be given to the multiple parties involved in the decision-making process. Reflective practice, opinion leadership and role modelling may offer ways forward but will require evaluation. PMID- 16045518 TI - Primary mode of delivery and subsequent pregnancy. AB - OBJECTIVE: To investigate the relationship between primary mode of delivery and subsequent pregnancy and to compare the findings with a previous study conducted on an earlier cohort from the same population. DESIGN: Population cohort. SETTING: Aberdeen City, Scotland. POPULATION: Women who delivered their first singleton child in Aberdeen Maternity Hospital between 1980 and 1997. METHODS: Population-based data relating to the index and next pregnancy event, if any, were obtained from the Aberdeen Maternity Neonatal Databank. Subsequent pregnancy was compared across the three modes of delivery groups using log rank tests and Cox proportional hazards regression models. MAIN OUTCOME MEASURE: First subsequent pregnancy following index delivery. RESULTS: Women who delivered by caesarean section (CS) were less likely to have a subsequent pregnancy compared with those who had a spontaneous vaginal delivery (SVD), hazard ratio = 0.91 (95% CI 0.87, 0.95). This confirmed the findings of a previous study conducted on an earlier cohort of the same population. The median time to next pregnancy following CS was 36.3 months, 31.8 months following instrumental vaginal delivery (IVD) and 30.4 months following SVD. In contrast to the earlier study where women who had an instrumental delivery were found to be an intermediate group, we found no difference in subsequent pregnancy following IVD compared with SVD, HR = 1.0 (95% CI 0.96, 1.03). CONCLUSIONS: Following an initial delivery by CS, fewer women went on to have another pregnancy compared with SVD. The incidence of subsequent pregnancy is similar following instrumental and SVD. PMID- 16045519 TI - Risk scoring system for prediction of obstetric anal sphincter injury. AB - OBJECTIVE: The objective was to begin the process of developing an antenatal risk scoring system, as a first step towards examining whether elective Caesarean section for women at high risk of injury could be an effective and acceptable intervention. DESIGN: Retrospective study. SETTING: Tertiary maternity unit in the UK. POPULATION: One hundred and twenty-three women who sustained an obstetric anal sphincter injury (OASI) and 123 controls without OASI. METHODS: Case notes of women with a third or fourth degree tear between 1997 and 1999 were examined for risk factors. Controls matched for age and week of delivery were identified from the maternity record database and case records reviewed for the presence of risk factors. Unweighted and weighted risk scores were produced using odds ratios, and compared between cases and controls. Receiver operating characteristics (ROC) curve analysis of the risk scores was performed to discriminate between cases and controls and to calculate the sensitivity and specificity of each scoring system. MAIN OUTCOME MEASURES: Odds ratio (OR) and 95% confidence interval (CI) for each risk factor. Sensitivity and specificity from ROC curves for weighted and unweighted risk score. RESULTS: Among the cases there were more nulliparous women (OR 1.77; CI 1.05-2.99) and a trend towards more women with an episiotomy (OR 1.57; CI 0.99-2.47). Among women with sphincter injury, trends towards more epidurals (OR 1.64; CI 0.97-2.75), and more babies weighing more than 4000 g among (OR 1.45; CI 0.85-2.49) were noted. The median unweighted risk score was 2 for cases and 2 for controls (P= 0.05), while the weighted risk score was 2.1 and 1.37 (P= 0.03), respectively. The ROC curves approximated to a straight line demonstrating very poor discrimination between cases and controls. CONCLUSION: The predictive test performed poorly, suggesting that the risk factors identified do not exert a large enough effect in a cohort of this size. PMID- 16045520 TI - Factors related to breast abscess after delivery: a population-based study. AB - OBJECTIVE: To investigate whether there are underlying factors, other than breastfeeding behaviours, which may contribute to the development of breast abscess during the year following delivery. DESIGN: A population-based study. SETTING: In Sweden. POPULATION: The 1,454,068 singleton deliveries during 1987 2000. Methods Data retrieval from two national patient registers: the Medical Birth Registry and the National Discharge Register. Stratified Mantel-Haenszel analysis. MAIN OUTCOME MEASURES: Odds ratios with 95% confidence intervals for possible risk factors for breast abscess in the year following delivery. RESULTS: During the year following delivery 1401 women had surgery because of a breast abscess. This gives a rate of 0.1%. Sixty-five percent of cases occurred between three and eight weeks postpartum. An annual increase in the odds ratio (OR) was found between 1993 and 1999. Lowest risk was found among mothers who were < or =24 years and a significantly increased risk among mothers > or =30 years. There was a 3.6-fold increased risk for breast abscess associated with primiparity and OR for post-maturity (>41 weeks) was 5. CONCLUSION: Primiparous women appear to be at a greater risk for the development of breast abscess during lactation than multiparous women. Mothers over the age of 30 years and those who give birth post maturely are also at a significantly increased risk. Primiparous women and women over 30 years could be targeted for extra information in preparation for parenthood classes about how to avoid over distension of the breasts during breastfeeding. Further research is needed to understand the role of post-maturity and to investigate why some women recover spontaneously from inflammatory processes of the breast. PMID- 16045521 TI - Faecal incontinence and mode of first and subsequent delivery: a six-year longitudinal study. AB - OBJECTIVE: To investigate the prevalence of persistent and long term postpartum faecal incontinence and associations with mode of first and subsequent deliveries. DESIGN: Longitudinal study. SETTING: Maternity units in Aberdeen, Birmingham and Dunedin. POPULATION: Four thousand two hundred and fourteen women who returned postal questionnaires three months and six years postpartum. METHODS: Symptom data were obtained from both questionnaires and obstetric data from case-notes for the index birth and the second questionnaire for subsequent births. Logistic regression investigated the independent effects of mode of first delivery and delivery history. MAIN OUTCOME MEASURES: Incontinence to bowel motions three months and six years after index birth. For delivery history, the outcome was incontinence only at six years. RESULTS: The prevalence of persistent faecal incontinence was 3.6%. Almost 90% of these women reported no symptoms before their first birth. The forceps delivery of a first baby was independently predictive of persistent symptoms (OR 2.06, 95% CI 1.40-3.04). A caesarean section first birth was not significantly associated with persistent symptoms (OR 1.07, 95% CI 0.64-1.81). Delivering exclusively by caesarean section also showed no association with subsequent symptoms (OR 1.04, 95% CI 0.72-1.50) but ever having forceps was significantly predictive (OR 1.48, 95% CI 1.18-1.87). Other factors independently associated with persistent faecal incontinence were older maternal age, increasing number of births and Asian ethnic group. Birthweight and long second stage were not significantly associated. CONCLUSIONS: The risk of persistent faecal incontinence is significantly higher after a first delivery by forceps. We found no evidence of a lower risk of subsequent faecal incontinence for exclusive caesarean section deliveries. PMID- 16045522 TI - Domestic violence risk during and after pregnancy: findings from a British longitudinal study. AB - OBJECTIVE: The objectives of this study were to examine the rates of domestic violence reported during and after pregnancy and to assess the importance of family adversity. DESIGN: Prospective longitudinal cohort study. SETTING: Bristol Avon, Southwest England. POPULATION: Seven thousand five hundred and ninety-one pregnant women with due dates between 1.4.91 and 31.12.92. METHODS: Questionnaires administered at 18 weeks of gestation and 8 weeks, 8 months, 21 months and 33 months postpartum. MAIN OUTCOME MEASURES: The experience of emotional or physical cruelty by an intimate partner at each time point. RESULTS: Fewer women reported domestic violence victimization during pregnancy than they did postpartum (18 weeks of gestation: 1% physical cruelty, 4.8% emotional cruelty, 5.1% any victimization; 33 months postpartum: 2.9% physical, 10.8% emotional, 11% any victimization). Women who reported being victimized during pregnancy also reported significantly higher levels of social adversity during pregnancy. The number of social adversities reported during pregnancy also predicted postpartum victimization. Women who reported only one adversity during pregnancy were 2.73 (95% CI, 2.16-3.45) times more likely to report physical victimization at 33 months postpartum. Women who reported 5 adversities during pregnancy were 14.69 (95% CI, 7.35-29.37) times more likely to report such victimization at 33 months postpartum. For emotional cruelty, women who reported only one adversity during pregnancy were 2.10 (95% CI 1.80-2.46) times more likely to report emotional victimization at 33 months postpartum and 6.10 (95% CI 3.51-10.59) times more likely to report such victimization when five or more adversities were present during pregnancy. CONCLUSIONS: Levels of social adversity reported in pregnancy are important predictors of concurrent and future victimization. Screening for social adversity factors could help identify women at high risk for future domestic violence. PMID- 16045523 TI - A randomised double blind trial comparing misoprostol or placebo in the management of early miscarriage. AB - OBJECTIVES: To study if misoprostol 400 microg, administered vaginally, increased the successful resolution of early miscarriage compared with placebo. DESIGN: Randomised, double blind placebo controlled study. SETTING: Sahlgrenska University Hospital, Goteborg, Sweden. SAMPLE: One hundred and twenty-six women seeking medical attention for early miscarriage. METHOD: Women with a non-viable, first trimester miscarriage were randomised to vaginal administration of misoprostol 400 microg or placebo. MAIN OUTCOME MEASURES: Main outcome measure was the proportion of successful complete resolution of miscarriage. Secondary outcomes were incidence of infection, bleeding, gastrointestinal side effects, pain, use of analgesics and length of sick leave between groups. RESULTS: Sixty four patients were randomised to misoprostol and 62 to placebo. Eighty-one percent in the misoprostol and 52% in the placebo group had a complete miscarriage within one week of the primary visit (RR 1.57; 95% CI 1.20-2.06). Patients in the misoprostol group reported more pain as assessed on a visual analogue scale (60.4 [31.0] vs 43.8 [37.1] mm; P < 0.007) and required analgesics more often (83%vs 61%, RR 1.35; 95% CI 1.08-1.70). There were no significant differences in the occurrence of gastrointestinal side effects, infection, reduction in haemoglobin or sick leave between the groups. CONCLUSIONS: Treatment with 400 mug misoprostol administered vaginally increased the success rate of resolvement of uncomplicated early miscarriages compared with placebo. However, women who received misoprostol experienced more pain and required more analgesics than those who did not. PMID- 16045524 TI - Modulation of cytokine production by dydrogesterone in lymphocytes from women with recurrent miscarriage. AB - OBJECTIVE: To examine the effects of dydrogesterone on the production of Th1 and Th2 cytokines by lymphocytes from women undergoing unexplained recurrent spontaneous miscarriage (RSM). DESIGN: Controlled prospective, clinical study conducted in a maternity hospital and a university-based immunology laboratory. SETTING: Faculty of Medicine, Kuwait University and Kuwait Maternity Hospital. SAMPLE: Thirty women with unexplained RSM. METHODS: Peripheral blood mononuclear cells (PBMC) from women with unexplained RSM were isolated from venous blood by density gradient sedimentation and stimulated with phytohaemagglutinin (PHA). Culture supernatants assayed for interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha, interleukin (IL)-4, IL-6 and IL-10 by ELISA. Levels of the progesterone-induced blocking factor (PIBF) were also measured. MAIN OUTCOME MEASURES: Cytokine production in the presence and absence of progesterone and dydrogesterone. RESULTS: Dydrogesterone significantly inhibited the production of the Th1 cytokines IFN-gamma (P= 0.0001) and TNF-alpha (P= 0.005) and induced an increase in the levels of the Th2 cytokines IL-4 (P= 0.03) and IL-6 (P= 0.017) resulting in a substantial shift in the ratio of Th1/Th2 cytokines. The effect of dydrogesterone was blocked by the addition of the progesterone-receptor antagonist mifepristone, indicating that dydrogesterone was acting via the progesterone receptor. Dydrogesterone induced the production of PIBF. CONCLUSION: Dydrogesterone inhibits the production of the Th1 cytokines IFN-gamma and TNF alpha from lymphocytes and up-regulates the production of the Th2 cytokines IL-4 and IL-6, inducing a Th1 to Th2 cytokine shift. PMID- 16045525 TI - A randomised controlled trial of mifepristone in combination with misoprostol administered sublingually or vaginally for medical abortion up to 13 weeks of gestation. AB - OBJECTIVE: To assess women's acceptability, the efficacy and side effects of sublingual versus vaginal administration of misoprostol in combination with mifepristone for medical abortion up to 13 weeks of gestation. DESIGN: Randomised controlled trial. SETTING: Aberdeen Royal Infirmary. POPULATION: Women undergoing medical abortion under the terms of the 1967 Abortion Act. METHODS: Mifepristone (200 mg) was given orally followed 36-48 hours later by misoprostol administration (sublingual: 600 microg; vaginal: 800 microg). A second dose of misoprostol 400 microg was given 3 hours later (sublingually or vaginally). Women between 9 and 13 weeks of gestation received a further (third) dose of misoprostol 400 microg (sublingually or vaginally), 3 hours later if abortion had not occurred. MAIN OUTCOME MEASURES: Women's acceptability, efficacy of the regimen and side effects experienced. RESULTS: A total of 340 women were recruited (171 sublingual and 169 vaginal). A total of 70% of women in the sublingual group expressed satisfaction with the route of misoprostol administration; 18% answered 'Don't know' while 12% were dissatisfied, compared with 68%, 28% and 4%, respectively, in the vaginal group (P= 0.02). There was no significant difference in the need for surgical evacuation for women in the sublingual (3/158, 1.9%) and vaginal groups (4/156, 2.6%) (P= 0.70). Women receiving misoprostol sublingually were more likely to experience diarrhoea (P < 0.01), shivering (P < 0.01) and unpleasant mouth taste (P < 0.01). CONCLUSIONS: Sublingual administration of misoprostol is an effective alternative to vaginal administration for medical abortion up to 13 weeks of gestation. The prevalence of prostaglandin-related side effects, however, was higher with this route of administration. PMID- 16045526 TI - A randomised controlled trial of microwave endometrial ablation without endometrial preparation in the outpatient setting: patient acceptability, treatment outcome and costs. AB - OBJECTIVE: To compare outpatient microwave endometrial ablation (MEA) in the postmenstrual phase to standard MEA treatment after drug preparation in a day case theatre environment. DESIGN: A randomised controlled trial. SETTING: A large United Kingdom teaching hospital. POPULATION: Two hundred and ten women complaining of excessive menstrual loss. METHODS: Two hundred and ten women with excessive menstrual loss were randomised. Ninety-seven women were treated as outpatients in the immediate post-menstrual phase and 100 were treated in an operating theatre after hormonal preparation. All procedures were commenced under local anaesthesia with or without conscious sedation. Analysis was by modified intention to treat. MAIN OUTCOME MEASURES: Primary outcome measures were satisfaction with treatment (measured at one year) and acceptability of treatment (measured at two weeks). Secondary outcome measures were menstrual outcome and financial cost. RESULTS: Significantly more women found treatment post-menses acceptable; 86 (89.5%) versus 76 (76.0%) [difference in proportions 13.6%, 95% CI (3.0%, 23.9%)]. Similar numbers in each arm were totally or generally satisfied with the treatment, 86 (92.5%) versus 84 (88.4%) [difference in proportions 4.1%, 95% CI (-4.7%, 12.9%)] while amenorrhoea rates at one year were comparable, 52 (55.9%) versus 60 (61.9%). [difference in proportions -5.9%, 95% CI (-19.8%, 7.6%)]. The mean health service costs were 124 pounds (95% CI 86-194 pounds) lower for the patients in the post-menses group. CONCLUSION: MEA performed under local anaesthesia (with or without conscious sedation) in the post-menstrual phase achieves high levels of satisfaction is very acceptable to patients and results in significantly reduced health service costs. Importantly menstrual outcomes are not affected by omission of drug preparation. There is now good evidence to support the use of MEA, without drug endometrial preparation, in the outpatient setting. PMID- 16045527 TI - Thermal ablation performed in a primary care setting: the South Warwickshire Experience. AB - OBJECTIVE: To assess the feasibility of performing outpatient thermal ablation in a primary care setting. DESIGN: Prospective study. SETTING: A small peripheral hospital in the UK used by local general practitioners and visiting hospital practitioners for its outpatient facilities. POPULATION: The area of South Warwickshire serves a population of 270,000. METHODS: Two general practitioners were trained to perform outpatient thermal ablation using the Thermachoice thermal ablation system (Thermachoice II). The unit functioned autonomously with support from a local gynaecologist and radiologist, accepting referrals from hospital consultants and general practitioners. MAIN OUTCOME MEASURES: Severity of menstrual loss, premenstrual symptoms, dysmenorrhoea and quality of life assessed by visual analogue scales before treatment and at one month, two months, one year and two years. RESULTS: Eighty-seven women were treated. No major complications were encountered from the procedure. Reduction of menstrual loss or cure was reported by over 94% of women. Premenstrual syndrome (PMS) and symptoms of dysmenorrhoea were also improved by treatment. The majority of women were satisfied with the operation at one month (96%), two months (93%), one year (92%) and two years (94%). CONCLUSIONS: Thermal ablation is a simple procedure well suited to an outpatient setting. There are few complications as a result of the use of the thermal ablation catheters, and this study has shown that the procedure can be undertaken in a primary care setting with excellent results. PMID- 16045528 TI - Randomised comparative trial of the levonorgestrel intrauterine system and mefenamic acid for the treatment of idiopathic menorrhagia: a multiple analysis using total menstrual fluid loss, menstrual blood loss and pictorial blood loss assessment charts. AB - OBJECTIVE: To compare the efficacy and tolerability of the levonorgestrel intrauterine system (LNG IUS) with mefenamic acid in the management of objective idiopathic menorrhagia. DESIGN: Phase III, Single centre, open, randomised, comparative, parallel group study. SETTING: District General Hospital in the United Kingdom. POPULATION: Fifty-one women with objective menorrhagia. METHODS: Twenty-five women randomised to receive the LNG IUS and 26 to oral mefenamic acid for six cycles. MAIN OUTCOME MEASURES: Change from baseline in menstrual blood loss (MBL), total menstrual fluid loss (TMFL) and pictorial blood loss assessment chart (PBAC) score at the third and sixth cycle of treatment. RESULTS: After six cycles the median menstrual blood loss was 5 mL in the LNG IUS group and 100 mL in the mefenamic acid group (P < 0.001). Median TMFL was 27 mL in the LNG IUS group and 157 mL in the mefenamic acid group (P < 0.001). Median PBAC score was 25 in the LNG IUS group and 159 in the mefenamic acid group. Changes in menstrual blood loss correlated strongly to changes in TMFL (r= 0.88) but PBAC correlated less well to blood loss and total fluid loss (r= 0.53 and r= 0.58). CONCLUSIONS: Both the LNG IUS and mefenamic acid significantly decreased menstrual blood loss, TMFL and PBAC scores. The LNG IUS produced greater reductions in all parameters than mefenamic acid. Comparison of the different measurements suggests that TMFL assessment may be an easier and a more relevant measure of symptom severity than menstrual blood loss. PMID- 16045529 TI - Socio-economic position across the life course and hysterectomy in three British cohorts: a cross-cohort comparative study. AB - OBJECTIVE: To examine the association between indicators of lifetime socio economic position and rates of hysterectomy in three British cohorts. DESIGN: Cross-cohort comparative study. SETTING: Two cohorts: England, Scotland and Wales. Third cohort: Aberdeen, Scotland. POPULATION: Three thousand two hundred and eight women born between 1919 and 1940, participating in the British Women's Heart and Health Study (BWHHS); 1394 women from the MRC National Survey of Health and Development (NSHD), followed up since birth in 1946; 3208 women born between 1950 and 1955, participating in the Aberdeen Children of the 1950s study, all with complete information on lifetime socio-economic position and hysterectomy status. METHODS: Relative indices of inequality were derived for markers of socio economic position in childhood and adulthood. Cox's regression models were used to test the association between these markers and hysterectomy. MAIN OUTCOME MEASURE: Self-reported hysterectomy with or without oophorectomy. RESULTS: Adverse socio-economic position in childhood and as indicated by educational status was associated with reduced rates of hysterectomy in the oldest of the three cohorts (BWHHS), whereas conversely in the NSHD and Aberdeen cohorts it was associated with increased rates of hysterectomy. The unadjusted hazards ratios for hysterectomy comparing worst to best socio-economic position for father's social class were 0.73 (0.56, 0.96) for women from the BWHHS, 1.77 (1.19, 2.65) for those from the NSHD and 2.06 (1.46, 2.89) for those from the Aberdeen cohort. Associations between markers of adult socio-economic position and hysterectomy tended to be weaker in all three cohorts and often did not reach conventional levels of statistical significance. CONCLUSIONS: Our results show that hysterectomy rates are influenced by childhood socio-economic position and educational attainment, but that the nature of this association varies across these three British cohorts born in different decades of the 20th century. That there were no consistent or strong associations between adult SEP and hysterectomy rates suggest that social factors influencing rates of hysterectomy are likely to be those experienced or which develop in early life rather than those which develop later. PMID- 16045530 TI - Long term review of laparoscopic sacrocolpopexy. AB - OBJECTIVE: Assessment of long term outcome following laparoscopic sacrocolpopexy. DESIGN: Retrospective follow up study using standardised examination with pelvic organ prolapse quantification system (POP-Q) and questionnaires. SETTING: A tertiary urogynaecology unit in the North West of England. POPULATION: One hundred and forty consecutive cases who had a laparoscopic sacrocolpopexy at St Mary's Hospital, Manchester, between 1993 and 1999. METHODS: Women completed questionnaires and were examined in gynaecology clinic or sent postal questionnaires if unable to attend the clinic. MAIN OUTCOME MEASURES: Adequacy of vault support and recurrent vaginal prolapse assessed by POP-Q score. Assessment of prolapse, urinary and bowel symptoms and sexual function using questionnaires. RESULTS: One hundred and three women were contacted after a median of 66 months. Sixty-six women were examined and a further 37 women filled in questionnaires only. Recurrent vault prolapse occurred in 4 of the 66 women who were examined. Prolapse had recurred or persisted in 21 of 66 women, with equal numbers of anterior and posterior vaginal wall prolapse. Overall, 81/102 (79%) said that their symptoms of prolapse were 'cured' or 'improved'; 39/103 (38%) still had symptoms of prolapse. For every two women who were cured of their urinary or bowel symptoms, one woman developed worse symptoms. CONCLUSIONS: Among the 66 women available for examination laparoscopic sacrocolpopexy provided good long term support of the vault in 92%. Forty-two percent of these women had recurrent vaginal wall prolapse. Despite this, 79% of women felt that their symptoms of prolapse were cured or improved following surgery. PMID- 16045531 TI - Mode of delivery and the risk of delivery-related perinatal death among twins at term: a retrospective cohort study of 8073 births. AB - OBJECTIVE: To determine the risk of perinatal death among twins born at term in relation to mode of delivery. DESIGN: Retrospective cohort study. SETTING: Scotland 1985-2001. POPULATION: All twin births at or after 36 weeks of gestation, excluding antepartum stillbirths and perinatal deaths due to congenital abnormality (n= 8073). METHODS: The outcome of first and second twins was compared using McNemar's test and the outcome of twin pairs in relation to mode of delivery was compared using exact logistic regression. MAIN OUTCOME MEASURES: Intrapartum stillbirth or neonatal death of either twin. RESULTS: Overall, there were six deaths of first twins and 30 deaths of second twins (OR for second twin 5.00, 95% CI 2.00-14.70). The odds ratio for death of the second twin due to intrapartum anoxia was 21 (95% CI 3.4-868.5). The associations were similar for twins delivered following induction of labour and for sex discordant twins. However, there was no association between birth order and the risk of death among 1472 deliveries by planned caesarean section. There was death of either twin among 2 of 1472 (0.14%) deliveries by planned caesarean section and 34 of 6601 (0.52%) deliveries by other means (P= 0.05, odds ratio for planned caesarean section 0.26 [95% CI 0.03-1.03]). The association was similar when adjusted for potential confounders. Assuming causality, we estimate that 264 caesarean deliveries (95% CI 158-808) would be required to prevent each death. CONCLUSION: Planned caesarean section may reduce the risk of perinatal death of twins at term by approximately 75% compared with attempting vaginal birth. This is principally due to reducing the risk of death of the second twin due to intrapartum anoxia. PMID- 16045532 TI - Bilateral iliococcygeal fixation for vaginal vault prolapse and enterocele repair using a new suturing device--the digital needle driver. AB - The objective of our study was to evaluate the surgical feasibility, efficacy and safety of the digital needle driver (DND 202), a modified, flexible surgical device, during iliococcygeal fixation (ICF) for vaginal vault prolapse and enterocele repair. A prospective longitudinal study was carried out among 21 consecutive patients who underwent bilateral iliococcygeal fixation at St George's Hospital, London. All patients filled a comprehensive questionnaire for pre- and post-operative prolapse, urinary, bowel and sexual symptoms and underwent pre- and post-operative site-specific vaginal examination, following the standardized International Continence Society scoring for prolapse, pre operative urodynamic studies and analysis of the surgical results. The outcome measures were the feasibility of the procedure, the time needed, intra- and post operative complications, short-term post-operative prolapse-associated symptoms and pelvic organ prolapse quantification. The mean age of the patients was 65 [5] years and the mean body mass index (kg/m(2)) was 23 [2.7]. In addition to ICF, 8 patients underwent vaginal hysterectomy, 18 had posterior repairs, 7 had anterior repairs and 6 had TVT. The mean time for ICF was 20 [11] minutes, the mean blood loss per surgical procedure was 264 [225] mL and the mean hospitalization time was 4.6 [1.2] days. Postoperatively, one patient had mesh erosion. At short-term post-operative evaluation none of the patient had prolapse symptoms. There was a statistically significant improvement in all stages of the apical and posterior walls prolapse (p < 0.001). The mean total vaginal length was significantly shorter postoperatively (7.8 [1.0] cm vs 6.6 [1.4] cm, p < 0.001). Thus, we can conclude that the use of DND device may facilitate the vaginal approach for vaginal vault prolapse and enterocele repair. PMID- 16045533 TI - Unusual presentation of Bartholin's gland duct cysts: anterior expansions. AB - The medical records of 32 patients who underwent surgery for a vulvar mucinous cyst located between the urethral meatus and the fourchette were reviewed. Nineteen (59%) patients had been referred to our centre because of cyst recurrence after incision or excision. The majority of the cysts were lined by a single layer of tall columnar cells with zones of squamous metaplasia. Subacute or chronic bartholinitis was demonstrated in 94% of the cases demonstrating that these cysts were anterior expansions of Bartholin's gland duct cysts. No ipsilateral recurrence was observed after cyst and gland excision. PMID- 16045534 TI - Peritoneal fluid concentrations of interleukin-17 correlate with the severity of endometriosis and infertility of this disorder. AB - This prospective study aimed to determine whether patients with endometriosis are having different level of interlukin-17 (IL-17) in peritoneal fluid when compared with patients without endometriosis. The patients with minimal/mild endometriosis had a significantly higher level of IL-17 in peritoneal fluid compared with those with moderate/severe endometriosis or without endometriosis. The concentration of IL-17 in peritoneal fluid was significantly higher when endometriosis and infertility coexist. However, the concentration of IL-17 in peritoneal fluid did not correlate with the phase of the menstrual cycle in the patients with or without endometriosis. Our study suggested that IL-17 might play an important role in the pathogenesis of early endometriosis and endometriosis-associated infertility. PMID- 16045535 TI - Are mediolateral episiotomies actually mediolateral? AB - This study investigated potential differences in the cutting of mediolateral episiotomy between doctors and midwives. Depth, length, distance from midline and shortest distance from the midpoint of the anal canal to the episiotomy were measured in a sample of primigravid women. The angle subtended from the sagittal or parasagittal plane was calculated. Two hundred and forty-one women participated of whom 98 (41%) had a mediolateral episiotomy. Doctors performed episiotomies that were significantly deeper, longer and more obtuse than those by midwives. No midwife and only 13 (22%) doctors performed truly mediolateral episiotomies. It appears that the majority of episiotomies are not truly mediolateral but closer to the midline. More focused training in mediolateral episiotomy technique is required. PMID- 16045536 TI - Neonatal death following termination of pregnancy. AB - The aim of this study was to look at neonatal death following termination of pregnancy in 31 cases over a six-year period in our region and to determine why this was occurring. We have highlighted two main areas of concern: failure to perform feticide in keeping with RCOG advice, and classification and registration below the clinical limit of viability. In these circumstances, appropriate counselling of the family and an antenatal plan for the postnatal care of the dying infant made prior to delivery are essential. We question the rationale of the existing classification and registration requirements below the clinical limit of viability. PMID- 16045537 TI - The effect of the Term Breech Trial on medical intervention behaviour and neonatal outcome in The Netherlands: an analysis of 35,453 term breech infants. PMID- 16045539 TI - Vitamin E for the treatment of dysmenorrhea. PMID- 16045540 TI - Dyspareunia following vaginal surgery for prolapse using polypropylene mesh. Re: paper by Milani et al. PMID- 16045542 TI - Overexpression of Fos-related antigen-1 in head and neck squamous cell carcinoma. AB - The activating protein-1 (AP-1) family of transcription factors has been implicated in the control of proliferation and differentiation of keratinocytes, but its role in malignant transformation is not clear. The aim of this study is to assess the pattern of mRNA expression of jun-fos AP-1 family members in 45 samples of head and neck squamous cell carcinomas (HNSCC) and matched adjacent mucosa by means of Northern blot analysis. Transcripts of all family members were identified, except for JunB that was detected only by means of reverse transcription polymerase chain reaction. Neither c-Fos nor JunD or FosB mRNA differed between tumours and normal tissues. We observed a strong Fos-related antigen-1 (Fra-1) and Fra-2 expression, but only Fra-1 mRNA densitometric values were higher in tumour, compared to normal adjacent mucosa (t-test, P = 0.006). A direct relationship between the positive expression of Fra-1 mRNA, above tumour median, was associated with the presence of compromised lymph nodes (Fischer exact test, P = 0.006). In addition, Fra-1 protein staining was assessed in a collection of 180 tumours and 29 histologically normal samples adjacent to tumours in a tissue array. Weak reactivity, restricted to the basal cell layer, was detected in 79% of tumour adjacent normal tissues, opposed to the intense reactivity of cancer tissues. In the subgroup of oral cancers, we have observed a shift in Fra-1 immunoreactivity, as long as the number of patients in each category, cytoplasmic or nuclear/cytoplasmic staining, was analysed (Fischer exact test, P = 0.0005). Thus, Fra-1 gene induction and accumulation of Fra-1 protein may contribute to the neoplastic phenotype in HNSCC. PMID- 16045541 TI - Dendritic cells in viral pathogenesis: protective or defective? AB - Dendritic cells (DC) are potent antigen-presenting cells that are critical in the initiation of immune responses to control and/or eliminate viral infections. Recent studies have investigated the effects of virus infection on the biology of DC. This review summarizes these changes, focusing on both the DC parameters affected and the viral factors involved. In addition, the central role of DC biology in the pathogenesis of several viral families, including herpesviruses, paramyxoviruses and retroviruses, is explored. The field of pathogen recognition by DC is addressed, focusing on its role in protecting the host from viral infection, as well as the ability of viruses to exploit such host receptor ligation and signalling to their replicative advantage. The hypothesis is proposed that virus and host have evolved a symbiotic relationship to ensure both viral transmission and host survival. PMID- 16045543 TI - Combined small-cell carcinoma of the stomach: p53 and K-ras gene mutational analysis supports a monoclonal origin of three histological components. AB - Primary small-cell carcinoma (SmCC) is extremely rare in stomach. We reported an autopsy case of combined gastric SmCC with p53 and K-ras mutational analysis. Histologically, the tumour was composed of well-differentiated adenocarcinoma surrounding the central dominant SmCC component with scattered nests of squamous cell carcinoma. Immunohistochemically, all the neoplastic components revealed strong expression for p53 protein. Based on these findings, we hypothesized that these histologically different components originated from a same progenitor cell that possessed p53 mutation. Using Laser-capture microdissection technique and mutational analysis, we identified the same point mutations of p53 gene (A-->G transversion in codon 239) and K-ras gene (G-->A transversion in codon 13) in all the neoplastic components, but not in the adjacent normal gastric epithelium. Our results strongly support a hypothesis that the combined SmCC of stomach in this case was of monoclonal origin. PMID- 16045544 TI - Mutual paracrine effects of colorectal tumour cells and stromal cells: modulation of tumour and stromal cell differentiation and extracellular matrix component production in culture. AB - Interactions of tumour and stromal cells influence tumour cell proliferation and differentiation, stromal cell phenotypic transdifferentiation and secretion of extracellular matrix (ECM) components. In this study, we established a monolayer and a three-dimensional cell-to-cell interaction model between canine mammary stromal cells and human colonic carcinoma cell lines (Caco-2 and HT-29) to investigate mutual paracrine effects of tumour cells and stromal cells on (i) tumour cell differentiation, (ii) production of ECM components and (iii) phenotypic transdifferentiation of stromal cells. We showed that when Caco-2 or HT-29 cells are cultured in collagen gels, they form a few small solid cell clusters with no lumina, but when cocultured with stromal cells, the tumour cells formed glandular structures with central lumina. This fibroblast-induced organization and differentiation of Caco-2 cells (not HT-29 cells) appeared to be mediated by transforming growth factor-beta (TGF-beta). Culturing of stromal cells, Caco-2 cells or HT-29 cells alone in both monolayers and gels resulted in weak tenascin-C expression in stromal cells and HT-29 cells and no expression in the Caco-2 cells. Coculturing of stromal cells with tumour cells resulted in increased tenascin-C expression in the stromal cells and HT-29 cells and induced expression of tenascin-C in the Caco-2 cells. This induction and increased expression of tenascin-C appeared to be mediated by TGF-beta. Culturing of stromal cells, Caco-2 cells or HT-29 cells alone on monolayers and in gels resulted in a weak expression of chondroitin sulfate (CS), chondroitin-6-sulfate (C-6-S) and versican in stromal cells and no expression in Caco-2 and HT-29 cells. Coculturing of stromal cells with tumour cells on monolayers and in gels resulted in increased CS, C-6-S and versican expression in stromal cells. This tumour cell-induced expression of CS, C-6-S and versican appeared to be mediated by TGF-beta and platelet-derived growth factor (PDGF). Coculturing of Caco-2 and HT-29 and stromal cells promoted the transdifferentiation of stromal cells into myofibroblasts, and this appeared to be mediated by TGF-beta. These results suggest that TGF-beta and PDGF are part of a paracrine system involved in stromal epithelial cell interaction important in stromal cell differentiation and ECM component production. PMID- 16045545 TI - Radiation sensitivities of 31 human oesophageal squamous cell carcinoma cell lines. AB - The purpose of this study was to determine the radiosensitivities of 31 human oesophageal squamous cell carcinoma cell lines with a colony-formation assay. A large variation in radiosensitivity existed among 31 cell lines. Such a large variation may partly explain the poor result of radiotherapy for this cancer. One cell line (KYSE190) demonstrated an unusual radiosensitivity. Ataxia telangiectasia-mutated (ATM) gene in these cells had five missense mutations, and ATM protein was truncated or degraded. Inability to phosphorylate Chk2 in the irradiated KYSE190 cells suggests that the ATM protein in these cells had lost its function. The dysfunctional ATM protein may be a main cause of unusual radiosensitivity of KYSE190 cells. Because the donor of these cells was not diagnosed with ataxia telangiectasia, mutations in ATM gene might have occurred during the initiation and progression of cancer. Radiosensitive cancer developed in non-hereditary diseased patients must be a good target for radiotherapy. PMID- 16045546 TI - Hyperproduction of fibrin and inefficacy of antithrombin III and alpha2 macroglobulin in the presence of bacterial porins. AB - Bacterial porins enhance the thrombin activity upon chromogen substrate chromozym. Should porin-dependent enhancement of thrombin activity take place also upon fibrinogen in vivo, this might greatly increase the fibrin production which, in turn, might lead to blood vessel obstruction. In this study, we demonstrate fibrin hyperproduction in a simplified coagulative system, consisting of fibrinogen and thrombin-pure molecules, in the presence of bacterial porins. In particular, bacterial porins, in the presence of thrombin, significantly increased the fibrin production compared with thrombin alone. Also, fibrin hyperproduction took place even in the presence of the thrombin inhibitors antithrombin III (AT III) or alpha2 macroglobulin (alpha2M). However, the thrombin-fibrinogen reaction in the presence of AT III or alpha2M did not generate fibrin, unless porins were present. In conclusion, porins not only enhance thrombin activity but also inhibit the antithrombin activity exerted by AT III or alpha2M. We hypothesize that, because of porins activity, fibrin is largely generated due to thrombin hyperactivation. Moreover, further fibrin is produced by thrombin, which is not blocked by two serpins for the presence of porins. These results might be relevant as to the occurrence of disseminated intravascular coagulation in sepsis by gram-negative bacteria, which are known to produce porins. PMID- 16045547 TI - Dietary copper supplements modulate aortic superoxide dismutase, nitric oxide and atherosclerosis. AB - The objective was to test the hypothesis that dietary copper inhibits atherosclerosis by inducing superoxide dismutase (SOD) and potentiating nitric oxide (NO). New Zealand White rabbits were fed either a cholesterol diet (n = 8) or a cholesterol diet containing 0.02% copper acetate (n = 8) for 13 weeks. We found that the intimal area was significantly smaller in the animals supplemented with copper (P < 0.005), although integrated plasma cholesterol levels were not significantly different. This was associated with a significant increase in aortic copper content (P < 0.05), SOD activity (P < 0.05) and Cu/Zn SOD mRNA (P < 0.05) and a significant decrease in nitrotyrosine content (P < 0.05). Furthermore, there was a positive correlation between aortic copper content and SOD activity (P < 0.005, R(2) = 0.83) and a negative correlation between aortic superoxide dimutase activity and nitrotyrosine content (P < 0.005, R(2) = 0.93). In organ bath experiments, the relaxation of precontracted carotid artery rings to calcium ionophore was greater in animals supplemented with copper. No difference in response to sodium nitroprusside was observed. These data suggest that in the cholesterol-fed rabbit, copper supplements inhibit the progression of atherosclerosis by increasing SOD expression, thereby reducing the interaction of NO with superoxide, and hence potentiating NO-mediated pathways that may protect against atherosclerosis. PMID- 16045548 TI - B-1 cells are pivotal for in vivo inflammatory giant cell formation. AB - The mechanisms that govern giant cell (GC) formation in inflammatory, neoplastic and physiologic conditions are far from being understood. Here, we demonstrate that B-1 cells are essential for foreign-body GC formation in the mouse. GCs were analysed on the surface of glass cover slips implanted into the subcutaneous tissue of the animals. It was demonstrated that GCs are almost absent on cover slips implanted into the subcutaneous tissue of BALB/c or CBA/N X-linked immunodeficient mice. As these animals do not have B-1 cells in the peritoneal cavity, they were reconstituted with B-1 cells obtained from cultures of adherent mouse peritoneal cells. Results showed that in B-1-reconstituted animals, the number of GCs on the implant surface surpassed the values obtained with preparations from wild animals. In animals selectively irradiated (pleural and peritoneal cavities) to deplete these cavities of B-1 cells, GCs were also not formed. Enriched suspensions of B-1 cells grown in culture were labelled with [(3)H]-tymidine and injected into the peritoneal cavity of naive mice before implantation of glass cover slips. After 4 days, about 17% of mononuclear cells had their nuclei labelled, and almost 70% of GCs had one or more of their nuclei labelled when analysed by histoautoradiographic technique. A few GCs expressed an immunoglobulin M when analysed by immunostaining and confocal microscopy. Overall, these data demonstrate that B-1 cells are pivotal in the mechanisms of foreign-body GC formation in the mouse. PMID- 16045549 TI - Biennial chances. PMID- 16045550 TI - Development of two search strategies for literature in MEDLINE-PubMed: nursing diagnoses in the context of evidence-based nursing. AB - PURPOSE: To develop and validate search filters for MEDLINE via PubMed according to two categories of the NLINKS-EBN matrix. METHODS: The search results of the search filters were compared to a gold standard. FINDINGS: The usage of nursing classification terms for the literature search in evidence-based nursing (EBN) is still limited because taxonomies are neither widely used in nursing literature nor applied for indexing by MEDLINE. The proposed filters achieved a sensitivity of 96% and a specificity of 94% for "secondary data" and a sensitivity of 87% and a specificity of 73% for "diagnostic tests." CONCLUSIONS: The usage of database specific search filters are a reliable and valid method to search for nursing classification terms in medical databases. PMID- 16045551 TI - Quality of nursing diagnoses: evaluation of an educational intervention. AB - PURPOSE: To investigate the effects on the quality of nursing diagnostic statements in patient records after education in the nursing process and implementation of new forms for recording. METHODS: Quasi-experimental design. Randomly selected patient records reviewed before and after intervention from one experimental unit (n = 70) and three control units (n = 70). A scale with 14 characteristics pertaining to nursing diagnoses was developed and used together with the instrument (CAT-CH-ING) for record review. FINDINGS: Quality of nursing diagnostic statements improved in the experimental unit, whereas no improvement was found in the control units. Serious flaws in the use of the etiology component were found. CONCLUSION. Nurses must be more concerned with the accuracy and quality of the nursing diagnoses and the etiology component needs to be given special attention. PRACTICE IMPLICATIONS: Education of RNs in nursing diagnostic statements and peer review using standardized evaluation instruments can be means to further enhance RNs' documentation practice. PMID- 16045552 TI - Recent trends of genitourinary endoscopy in children. AB - Downsizing and refinement of the pediatric endoscope in video-monitoring systems have facilitated genitourinary endoscopy even in small children without any traumatic instrumentation. Indications for endoscopy in children with hematuria or tractable urinary tract infection have been tailored for the rareness of genitourinary malignancy or secondary vesicoureteral reflux (VUR) as a result of infravesical obstruction. Most mechanical outlet obstructions can be relieved endoscopically irrespective of sex and age. Endoscopic decompression by puncture or incision of both intravesical and ectopic ureteroceles can be an initial treatment similar to open surgery for an affected upper moiety. Endoscopy is necessary following urodynamic study to exclude minor infravesical obstruction only in children with unexplained dysfunctional voiding. Genitourinary endoscopy is helpful for structural abnormalities before and at the time of repairing congenital urogenital anomalies. Endoscopic injection therapy of VUR has been established as a less invasive surgical treatment. Pediatric endoscopy will play a greater role in the armamentarium for most pediatric urological diseases through the analysis of visual data and discussion on the indications for endoscopy throughout the world. PMID- 16045553 TI - Effect of tamsulosin on the number and intensity of ureteral colic in patients with lower ureteral calculus. AB - BACKGROUND: We aimed to objectively determine whether tamsulosin as an alpha(1) blocker was effective in patients who had ureterolithiasis located in the lower part of the ureter. METHODS: Sixty patients with lower ureteral calculi (juxtavesical or intramural portion) were included in the present study. Conservative treatment, such as hydration and tenoxicam as a non-steroidal anti inflammatory drug, was given to group 1 (30 patients). Group 2 (30 patients) was given tamsulosin (0.4 mg daily) in addition to the conservative treatment. All patients were followed up and questioned about the numbers and intensity of ureteral colic, and the rates of spontaneous passage after the procedure. RESULTS: Spontaneous passage was observed in 22 of the 30 patients in group 1 (73.3%) and 26 of the 30 patients in group 2 (86.6%). The difference within groups 1 and 2 was not significant (P=0.196). The difference between both groups was not statistically significant either, with the stone diameter being 6 mm (P=0.635) or >6 mm (P=0.407). As group 1 patients were passing their stones, they had more ureteral colic episodes than group 2 patients. This difference was statistically significant and correlated well with the administration of tamsulosin (P=0.038). Group 1 patients reported higher scores according to a visual analog scale than group 2 patients. Also, this difference was statistically significant (P=0.000). CONCLUSIONS: We think that the treatment of alpha(1)-blockers decreased the number of ureteral colic episodes and the intensity of pain during spontaneous passage at the lower ureteral calculi. Also, it will be beneficial to patients' quality of life. PMID- 16045554 TI - Questionnaire survey on female urinary frequency and incontinence. AB - BACKGROUND: Urinary incontinence is a well-known bothersome symptom in women, which may cause physical and psychological problems. We conducted a questionnaire survey on female urinary incontinence to investigate the disease's impact on the quality of life (QoL), the reasons women don't seek medical attention and the information they wished to obtain. PATIENTS AND METHODS: From March to October 2002, a member of the Professional Women's Coalition for Sexuality and Health distributed a questionnaire about urinary frequency and urinary incontinence to women who were attending the lectures hosted by the group. RESULTS: We analysed 262 questionnaires: 158 people belonged to the Stress Urinary Incontinence (SUI) Group, 36 to the Overactive Bladder (OAB) Group, 22 to the Urinary Frequency (UF) Group and 18 to the Normal Group. 'Going out' was most influenced in all three groups with symptoms in their daily life. Compared with the Normal or SUI Group, the QoL in those belonging to the OAB and UF Groups was more deteriorated. The medical institution check-up rate in the SUI Group was the lowest at 7.1%, bringing down the overall consultation rate to 13.5%. More than 70% of respondents who didn't have a check-up said that they did not think it was a problem serious enough to require consultation. CONCLUSIONS: Urinary incontinence and frequency impairs women's QoL. It is important to provide information on these diseases and to provide medical treatments that cater to the needs of individual patients. PMID- 16045555 TI - Urinary control after the definitive reconstruction of cloacal anomaly. AB - PURPOSE: Urinary control after definitive repair of a cloacal anomaly is difficult to achieve. The present report aims to describe the clinical course of urinary control, and the need for the management of bladder dysfunction after reconstruction. METHODS: The present consecutive series consisted of 11 girls who underwent definitive repair of cloacal anomalies over a period of 11 years. Eight patients were associated with hydrocolpos. Radiological examination included a plain X-ray radiograph of the lumbosacral spine and a voiding cystourethrography with or without a urodynamic study. RESULTS: Reconstruction of the cloaca was performed on patients aged between 1 and 3 years using a posterior sagittal approach. Vaginal reconstruction was carried out 13 times in 11 patients using tubularized vaginal flap, distal rectal segment, perineal skin flap, or total urogenital sinus mobilization. Cystostomy or vesicostomy was carried out in four newborns/infants. Another seven patients could void spontaneously but incompletely with residual urine. Occult spinal dysraphism was found in five patients and hemisacrum in two patients. After definitive reconstruction, most patients acquired an adequate to normal bladder volume for 1-year-olds. Normal detrusor-sphincter function was seen in three patients. Detrusor areflexia was seen in two patients who underwent in utero vesico-amniotic shunt. Detrusor underactivity was observed in six patients. Bladder compliance was good in all patients except for one. No patients in the present series showed persistent urinary incontinence from the bladder neck or urethral dysfunction. CONCLUSION: It is postulated that wetting after definitive repair may be the result of overflow incontinence and poor bladder contractility rather than sphincter injury. The main clinical characteristic of bladder dysfunction was a failure to empty. We could not define the exact etiology, but iatrogenic injury from extensive dissection can lead to the higher risks of peripheral nerve damage. Accomplishment of definitive repair involves not only anatomical reconstruction, but also postoperative urinary control, including the initiation of clean intermittent catheterizations under repeated urodynamic evaluations. PMID- 16045556 TI - Determination of gelatinase A using a modified indirect hemagglutination assay in human prostate cancer screening and assessment of its correlation with prostate specific antigen parameters. AB - BACKGROUND: Prostate cancer is the most common malignancy affecting men and is a major cause of cancer death. There are increasing data on novel tumor markers, such as gelatinase A, which play a key role in tissue invasion and metastasis. OBJECTIVES: We designed a study to evaluate total gelatinase A content using a simple and applicable Indirect hemagglutination (IHA) test in harmony with gelatinase A activity in serum samples as compared with prostate-specifc antigen (PSA) parameters. METHODS: In this study, we analysed the circulating form of gelatinase A (MMP-2) in patients suffering from either benign prostate hyperplasia (n=54) or prostate cancer (n=26) versus normal individuals as control (n=26). The gelatinolytic activity was determined by zymography and total MMP-2 content was measured by a novel IHA method. Total PSA and free PSA were quantified using a standard ELISA technique. RESULTS: Correlation of densitometric analysis of gelatinase A activity and IHA titer is significant at the 0.01 level (P<0.01, rho=0.916). Correlation of PSA and IHA titer is significant at the 0.01 level (P<0.01, rho=0.746). Correlation of free PSA and IHA titer is significant at the 0.01 level (P<0.01, rho=0.749). Borderline of IHA titer in patients with prostate cancer was 512+/-1 tube titer, in benign prostate hyperplasia patients was 128+/-1 tube titer and the titer in normal individuals was 8+/-1 tube titer. CONCLUSIONS: These results demonstrate that assessment of gelatinase A might be a promising procedure for monitoring and screening patients with prostate cancer. PMID- 16045557 TI - Trends in prostate cancer incidence and survival in various socioeconomic classes: a population-based study. AB - OBJECTIVES: Prostate cancer is currently the commonest cancer in men of all ages in UK, but robust demographic data of its distribution in various socioeconomic classes is lacking. We aimed to analyze its incidence, mortality and survival trends in West Midlands, England, from 1986 to 2000 in terms of socioeconomic deprivation. METHODS: Data were collated from the regional cancer registry database and a well-validated demographic score, the Townsend band, was employed as an indicator of social deprivation, including four variables as proxy indicators of socioeconomic status. Individual cases were allocated to one of five deprivation categories using postcode at diagnosis. Regression trend analysis at 1 and 5 years was performed and a P-value derived from the t-test statistic. RESULTS: In the mid-1980s, the incidence rate ratio in affluent:deprived classes was 0.9, with age-standardized rates of 35.23 and 39.53 per 100 000 people. This ratio increased to 1.5 by 2000 with age-standardized rates of 95.98 and 63.13, respectively (172% increase in affluent compared with 60% in deprived). The affluent groups had a 7 and 13% survival advantage at 1 and 5 years; the survival advantage at 1 year was statistically significant (P=0.03). CONCLUSIONS: The preferential changes in incidence and survival in the affluent social classes are likely to be due to heightened awareness, resulting in increased prostate-specific antigen testing, which captures early and relatively slow-growing tumors with a better overall prognosis. If these bipolar trends are allowed to persist, then the gap between the affluent and deprived will continue to widen. PMID- 16045558 TI - Expression of sialylated MUC1 in prostate cancer: relationship to clinical stage and prognosis. AB - AIM: MUC1 is distributed among a variety of normal epithelial tissues, and overexpression of MUC1 is detected in several human cancers. This study aimed to elucidate whether sialylated MUC1 expression correlated with: (i) clinical stage of prostate cancer; (ii) pathological grade of prostate cancer; (iii) pretreatment serum level of prostate-specific antigen (PSA); or (iv) the disease prognosis in patients with prostate cancer who received endocrine therapy. METHODS: We collected 57 biopsy specimens from prostate cancer patients treated with only endocrine therapy, and 10 specimens of normal prostates. These specimens were stained immunohistochemically by using a novel monoclonal antibody, MY.1E12, to detect sialylated MUC1. The levels of expression, clinical stages, pathological grades, pretreatment serum level of PSA and the prognoses of the patients were statistically analyzed for correlations. RESULTS: There were statistically significant correlations between the expression of sialylated MUC1 and pathological grades (WHO grade, P<0.01; Gleason score, P<0.05). Expression increased according to the progression of the disease (existence of clinical metastasis, P<0.05; clinical T-stage, P<0.01). Patients with high serum levels of PSA had higher expression than those with low levels (P<0.01). The level of sialylated MUC1 significantly correlated with progression-free survival (P<0.01) and cause-specific survival (P<0.01) according to univariate analyses. Furthermore, the level significantly correlated with progression-free survival according to multivariate analysis. CONCLUSIONS: These results suggest that sialylated MUC1 plays an important role in the progression of prostate cancer, and that its expression level in the primary lesion is a useful marker for the prognoses of patients undergoing endocrine therapy. PMID- 16045559 TI - Clinical characteristics of prostate cancer in Japanese men in the eras before and after serum prostate-specific antigen testing. AB - BACKGROUND: We retrospectively reviewed a large series of Japanese men with histologically proven prostate cancer to assess the clinical characteristics of the cancer in three different eras of prostate cancer management since prostate specific antigen (PSA) testing started in 1988. METHODS: The medical records of 1125 patients treated between 1975 and 2002 were reviewed with respect to age, chief complaints, clinical stage, tumor grade, treatment options at each stage, and prognosis. We classified the patients as follows: those treated in the pre PSA era between 1975 and 1988 (n=182), those treated in the PSA era between 1988 and 1997 (n=301; PSA era phase 1) and the PSA era between 1998 and 2002 (n=642; PSA era phase 2). RESULTS: Compared with the pre-PSA era, there were significant increases in the proportion of well-differentiated adenocarcinoma with respect to the biopsy tumor grade (24 vs 35%, P<0.01), in the proportion of linically organ confined disease (21 vs 43%, P<0.001), and in the proportion of patients who underwent radical prostatectomy (13%vs 20%, P<0.01) after PSA testing was introduced. In addition, there was a significant difference in the proportion of subjects who were 70--79 years of age between the pre-PSA era (52%, 95/182) and the PSA era phase 2 (42%, 270/642, P<0.05). There was also a significant difference in the proportion of patients who underwent surgical castration between the pre-PSA era (78%) and PSA era phase 2 (10%, P<0.001). The proportion of patients participating in prostate cancer screening increased from 3% (pre-PSA era) to 11% (PSA era phase 1 and PSA era phase 2, P<0.05). In all clinical stages, there were significant differences between the pre- and post-PSA eras in cause-specific survival rates (5-year: 74 vs 94% in stages A and B, P<0.01; 54 vs 89% in stage C, P<0.001; 32 vs 53% in stage D, P<0.001). CONCLUSIONS: Migrations in the age of patients (toward younger patients), the stage of the cancer (towards earlier stages) and the histological findings (toward favorable findings), in addition to changes in treatment options, have contributed to the prolonged survival of Japanese men with prostate cancer after the PSA testing was introduced. PMID- 16045560 TI - Accuracy of ultrasonography and magnetic resonance imaging in the diagnosis of non-palpable testis. AB - BACKGROUND: Studies were needed to evaluate the accuracy of ultrasonography (US) and magnetic resonance imaging (MRI) for diagnosing non-palpable testis, and to study the clinical usefulness of US and MRI before surgical exploration. METHODS: Fifty-six patients with non-palpable testis aged 1--12 years were examined between 1993 and 2002. Forty-six patients (55 non-palpable testes) underwent US. Forty patients (47 non-palpable testes) were examined using MRI. Both US and MRI were used in 29 patients (34 non-palpable testes). We used Fisher's exact probability test to compare the results of these graphic examinations with the surgical or laparoscopic findings for the non-palpable testis. RESULTS: Ultrasonography had a sensitivity of 76%, a specificity of 100%, and an accuracy of 84% in the diagnosis of non-palpable testis, whereas MRI had a sensitivity of 86%, a specificity of 79%, and an accuracy of 85%. CONCLUSIONS: There were no statistically significant differences in the accuracy rates of US and MRI for diagnosing non-palpable testis. Laparoscopy or inguinal exploration should be performed even if these graphic examinations demonstrate the absence of a testis, because some patients with a non-palpable testis were found to be false negatives. We recommend US as the first step after clinical examination, and MRI could be performed when the US findings are negative preoperatively for a non palpable testis. PMID- 16045561 TI - Molecular evaluation of the SRY gene for gonads of patients with mixed gonadal dysgenesis. AB - AIM: To determine whether the SRY gene is present in the gonads of patients with mixed gonadal dysgenesis (MGD). METHODS: Molecular analysis was performed in three patients with MGD. Polymerase chain reactions were used to test for the presence of the SRY gene in the peripheral lymphocytes, testes and streak gonads. RESULTS: Chromosome analysis revealed 45,XO/46,XY in two patients, and 46,XY in the third patient. In the peripheral lymphocytes and testicular tissue, the SRY sequences were positive in all cases. However, the SRY sequence was detected in the streak gonad in only two of the three patients. CONCLUSIONS: It is interesting that we identified both SRY-positive and SRY-negative streak gonads. Although the SRY gene has a very important role in testicular differentiation, genes other than the SRY gene might also influence the development of the indifferent gonad in MGD. PMID- 16045562 TI - Effect of urothelium on bladder contractility in diabetic rats. AB - AIM: It is known that physiopathological changes in diabetes affect the function of the bladder. In this study, we aimed to demonstrate the possible effects of diabetes on the urothelium during this physiopathological process. METHODS: Diabetes was induced in rats by tail vein injection of 35 mg/kg streptozotocin. Eight weeks later, intact and denuded bladder strips were prepared from these rats. Electrical field stimulation (EFS; 0.5-32 Hz), carbachol (10(-8)-10(-3) mol/L; cumulative dosage-response curves) and KCl (120 mmol/L) were used for the evaluation of the contractile responses. All responses were expressed as mg tension developed per mg of bladder tissue. Weights of rats and of their bladders, blood glucose levels, and frequency- and concentration-response curves were compared using anova, the paired t-test and the independent t-test. Differences were considered significant at P<0.05. RESULTS: Although no differences related to the weight of bladders of the control and diabetic groups were observed, there were differences in blood glucose levels and body weights between the two groups. Similarly, although there were no differences between the data obtained with EFS and KCl from tissues with intact and denuded strips in the control group, carbachol responses significantly differed between intact and denuded strips in the non-diabetic group. These differences were not observed in the diabetic group. In the control groups, in the presence of additional strips with intact urothelium placed in the medium containing denuded tissue, the differences in contractile responses between the intact control strip and the denuded strip disappeared. CONCLUSIONS: Diabetes possibly changes the interaction between the relaxant factors that are released from urothelium and muscarinic stimulation, but these interactions are not completely understood yet. Consequently, the response of the bladder to contractile stimulants is also affected. Further studies are required to reveal the mechanism by which diabetes influences the urothelium. PMID- 16045563 TI - Renal artery pseudoaneurysm after laparoscopic partial nephrectomy for renal cell carcinoma in a solitary kidney. AB - Renal artery pseudoaneurysms are a well-documented complication following trauma or percutaneous urological procedures, but are rare after partial nephrectomy. We present the case of a 34-year-old woman who, after undergoing a left nephrectomy in childhood due to Wilms' tumor, had a pseudoaneurysm in a solitary kidney after laparoscopic right partial nephrectomy with extraperitoneal approach for a renal cell carcinoma. The segmental renal artery feeding the pseudoaneurysm was embolized with coils without significant loss of residual renal function. PMID- 16045564 TI - Combined small cell carcinoma and sarcomatoid squamous cell carcinoma in the renal pelvis. AB - We report here a case of combined small cell carcinoma and sarcomatoid squamous cell carcinoma in the renal pelvis. A 61-year-old female presented with right flank discomfort, microhematuria and progressive renal dysfunction. Following diagnosis of right renal pelvic carcinoma, radical nephroureterectomy with lymph node dissection was performed through a midline incision. The tumor was pathologically diagnosed to be combined small cell carcinoma and sarcomatoid squamous cell carcinoma in the renal pelvis. The patient had no evidence of recurrence or metastasis, 16 months postoperatively. Small cell carcinoma or sarcomatoid squamous cell carcinoma of the renal pelvis is very rare. We believe this is the first such case to be reported in the world. PMID- 16045565 TI - Urinary tract involvement in a child with epidermolysis bullosa simplex. AB - Epidermolysis bullosa is a rare disorder which may present various systemic involvement. We present a 15-month-old boy with epidermolysis bullosa simplex who was admitted with symptoms of hematuria and dysuria and discuss the pathology from a urological standpoint. PMID- 16045566 TI - Endoscopic treatment of multiple prostatic calculi causing urinary retention. AB - Although prostatic calculi are relatively common, the etiology of these stones is not clear. We report a case with multiple prostatic calculi causing urinary obstruction and a concomitant bladder stone. We treated these stones endoscopically. We found a lot of different sized stones endoscopically, some protruding into the urethra, some filling different cavities on the prostate. So these cavities suggest prostatic calculi may occur related to intraprostatic reflux in the congenital or acquired diverticulum of the prostatic tissue. In addition, the stone composition of the bladder and prostatic stones was the same. All of these results show that the origin of bladder and prostatic stones can be the same. This case also supports a theory of intraprostatic reflux and urine stasis. PMID- 16045567 TI - Intravesical tape erosion following the tension-free vaginal tape procedure for stress urinary incontinence. AB - We report a case of intravesical tape erosion which occurred 6 months after the tension-free vaginal tape (TVT) procedure, which was successfully treated by partial tape removal. A 75-year-old woman, who had complained of recurrent cystitis after TVT procedure, was referred to Kobe City General Hospital in August 2003. The intravesical tape erosion was revealed by cystoscopy and computed tomography, and the tape was removed with a retropubic approach. The recurrent cystitis has been cured and the stress urinary incontinence has not recurred. The intravesical tape erosion was rare, with only seven reported cases including the present case. PMID- 16045568 TI - Surgical management of vaginal wound healing defects after tension-free vaginal tape placement. AB - The use of tension-free vaginal tape (TVT) made of polypropylene mesh carries the risk of causing vaginal wound healing defects, which are usually detected within 4 weeks of operation. We present three cases of vaginal wound healing defects after TVT procedures. These defects were managed using surgical techniques aimed at preserving the mesh, but all failed. There are many options for dealing with the problems of vaginal wounds following TVT. These include further observation, mesh removal, suturing the wound, or using tissue flaps to cover it. However, the latter two options cannot be recommended as they usually fail. PMID- 16045569 TI - Complete duplication of the bladder, urethra and external genitalia in a male neonate with an imperforate anus. AB - A rare case of a male neonate with complete duplication of the bladder, urethra and external genitalia with associated anomalies is reported. In addition to the above, the patient had a hypoplastic left kidney, bilateral bifid scrotums (four hemi-scrotums), an imperforate anus and a mass on the perineum. PMID- 16045570 TI - Severe ischemia of the glans penis following circumcision: a successful treatment via pentoxifylline. AB - Severe ischemia or necrosis of glans penis is rare. We report the case of an 11 year-old boy with severe glanular ischemia occurring 24 h after circumcision. This was successfully treated with pentoxifylline injection for 5 days, and while the black color of the glans penis changed to brownish at 48 h, appearances were close to normal at 5 days. The patient did not require any surgical intervention, and was discharged without sequelae. We suggest that pentoxifylline might be considered as a treatment of choice for severe ischemia of glans penis. PMID- 16045571 TI - Abstracts of the 35th Annual Scientific Meeting of the Australian and New Zealand Society of Nuclear Medicine, Melbourne, Australia, 30 April-2 May 2005. PMID- 16045572 TI - Stent insertion or endoluminal brachytherapy as palliation of patients with advanced cancer of the esophagus and gastroesophageal junction. Results of a randomized, controlled clinical trial. AB - SUMMARY: Esophageal cancer often presents as advanced stage disease with a dismal prognosis, with only 10-15% of patients surviving 5 years. Therefore, in a large proportion of patients, palliative treatment is the only option available. The aim of this study was to prospectively compare the palliative effect of self expandable stent placement with that of endoluminal brachytherapy regarding the effect on quality of life and on specific symptoms. Sixty-five patients with advanced cancer of the esophagus or gastroesophageal junction were randomized to treatment with either an Ultraflex expandable stent or high-dose-rate endoluminal brachytherapy with 7 Gy x 3 given in 2-4 weeks. Clinical assessment and health related quality of life (HRQL) were measured at inclusion and 1, 3, 6, 9 and 12 months later. The HRQL was measured with standardized questionnaires (European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core 30, European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Oesophageal Module and Hospital Anxiety and Depression Scale). Twenty-eight patients completed the stent treatment and 24 patients the brachytherapy. The group of patients treated with stent reported significantly better HRQL scores for dysphagia (P < 0.05) at the 1-month follow-up, but most other HRQL scores, including functioning and symptom scales, deteriorated. Among brachytherapy-treated patients, improvement was found for the dysphagia-related scores at the 3-months follow-up, whereas other significant changes of scores were few. The median survival time was comparable in the two groups (around 120 days). In conclusion, insertion of self-expandable metal stents offered a more instant relief of dysphagia compared to endoluminal brachytherapy, but HRQL was more stable in the brachytherapy group. PMID- 16045573 TI - Nonesophageal side-effects after antireflux surgery plus acid-suppression duodenal diversion surgery in patients with long-segment Barrett's esophagus*. AB - SUMMARY: During the last years we have employed acid-suppression duodenal diversion procedures (truncal vagotomy-partial gastrectomy plus Roux-en-Y gastrojejunostomy) in addition to antireflux surgery in order to treat all the pathophysiological factors involved in the genesis of Barrett's esophagus. We have observed very good results concerning the clinical and objective control of GERD at the long-term follow up after this procedure. However, it could be associated with other nonesophageal symptoms or side-effects. This study was conducted to evaluate the presence of gastrointestinal symptoms (diarrhea, vomiting, dumping, weight loss and anastomotic ulcers) after this operation. In this prospective study 73 patients were assessed using a careful clinical questionnaire asking regarding these complications at the early (< 6 months) and late (> 6 months) follow-up (average of 32.4 months). In the early postoperative period, diarrhea was present in 64% (19% considered severe 10-90 days after surgery), dumping in 41% and loss of weight in 71% of cases. Diarrhea occurred daily in 47.7% in the early postoperative period, but only in 16% of cases after 1 year. Shortly after surgery, steatorrea was observed in 9% of cases and responded well to medical treatment. Severe diarrhea or dumping was rare (5% of cases). These symptoms improved significantly after 1 year with medical management (45%, 20% and 30%, respectively) and 42% of patients regained their normal body weight. Only two patients presented anastomotic ulcers and were treated satisfactory with proton pump inhibitors. Revisional surgery was indicated in two patients with severe dumping syndrome. Most side-effects identified by this study were mild and diminished 1 year after operation. PMID- 16045574 TI - Elevated serum levels of C-reactive protein are indicative of a poor prognosis in patients with esophageal cancer. AB - SUMMARY: Despite improved results gained by the combination of neoadjuvant chemotherapy and radiotherapy with surgery, the prognosis for the esophageal carcinoma patient remains poor. There is a need for prognostic markers that can help improve patient selection. The aim of this study was to evaluate whether serum levels of C-reactive protein (CRP) have any influence on the patient prognosis in terms of prediction of chemoradiosensibility, and overall and disease-free survival. Blood samples were obtained at the time of diagnosis in 67 patients with biopsy-proven primary carcinoma of the esophagus. Although not significantly, the mean CRP level was higher than that of 20 patients with benign esophageal pathology who served as controls. No significant correlation could be found between CRP levels and baseline characteristics such as age, gender, histology, cancer localization, tumor depth, lymph node status or metastatic spread. However, patients with CRP levels higher than 6 mg/L were more frequently non-responders to chemoradiotherapy (P = 0.035), had a shorter overall survival (P = 0.061) and a shorter disease-free survival (P = 0.016). It is concluded from these results that pretreatment measurement of serum CRP levels in patients with esophageal cancer could to be used in routine practice as indicators of chemoradiosensibility and prognosis. PMID- 16045575 TI - Use of low dose dopamine, gabexate mesilate and ulinastatin reduces the water balance and pulmonary complication in thoracic esophagectomy patients. AB - SUMMARY: In spite of improvements in surgical technique and perioperative care, severe lung complication remains as the leading cause of morbidity in thoracic esophageal cancer patients who undergo esophagectomy. The purpose of this study was to evaluate the safety and effectiveness of postoperative drug therapy using low dose dopamine, gabexate mesilate, and ulinastatin on postoperative lung complication in esophageal cancer patients. Sixty-one patients operated for esophageal cancer from 1996 to 2000 were treated postoperatively with low dose dopamine (300 microg/kg/h), gabexate mesilate (80 mg/h), and ulinastatin (300 000 unit/day) as a study group. Seventy-four patients operated from 1987 to 1994 served as an historical control group. Various preoperative and perioperative medical parameters and water balance were analyzed. Postoperative pulmonary complications were observed in 26 patients (35.1%) in the control group and three patients (4.9%) in the study group, respectively (P < 0.0001). Preoperative and perioperative variables were not significantly different between the groups. Water balance from operation to postoperative day 3 in the study group was significantly lower than the control group. Postoperative use of low dose dopamine, gabexate mesilate, and ulinastatin significantly reduced pulmonary complications after esophagectomy. This may be partly attributable to negative water balance during the early postoperative days. PMID- 16045576 TI - Does transthoracic compared to transhiatal resection alter the early postoperative course of oesophagectomy? AB - SUMMARY: Several techniques for esophageal resections have been reported. However, clear clinical evidence is still lacking whether any of the procedures is superior to the others regarding morbidity and mortality in the early postoperative period. Two operative approaches for esophageal carcinoma, transhiatal and transthoracic, were compared with respect to operative morbidity, mortality and systemic inflammatory response. In our prospective study between 2000 and 2002 83 patients were investigated. In a retrospective post hoc analysis patients were divided into two groups due to the performed operational procedure, transhiatal (TH) or transthoracic (TT). Multiple Organ Dysfunction Score (MODS) was monitored daily (t1,t2,t3). Serum procalcitonin (PCT) levels were determined on admission to the ICU (t0), then 24 hourly (t24,t48,t72). Microalbuminuria (M:Cr) was measured before (tp), and after surgery (t0,t6,t24,t48,t72). For statistical analysis Wilcoxon rank sum test, Mann-Whitney U-test and chi-square test were used as appropriate. We examined 52 patients in the TH group, and 31 patients in the TT group. There was no significant difference between the two groups regarding age, male/female ratio, and SAPS II scores. Operations lasted significantly longer in the TT group: 375 (300-480) min compared to the TH group 240 375 (180-319) min, P < 0.001. ICU mortality was similar in both groups (TH: 46 survivors/6 non-survivors; TT: 27 survivors/4 non-survivors; P = 0.607, respectively). Daily MODS did not differ significantly between the two groups. The observed inflammatory markers (PCT and M:Cr) followed the pattern we described earlier, without significant difference. In this study there was no difference in the clinical and biochemical variables of the patients between the transthoracic and transhiatal groups. The observed early postoperative morbidity and mortality may indicate, that both procedures can be considered as procedures with similar postoperative risk. PMID- 16045577 TI - The effect of esophageal myotomy and myectomy on the lower esophageal sphincter pressure profile: intraoperative computerized manometry study. AB - SUMMARY: The surgical treatment of achalasia, based on Heller's myotomy is the procedure of choice to reduce the sphincterial high pressure zone, either by laparotomy or, most recently, by laparoscopy. What is the right length of the myotomy? Many authors have reported 10-15% postoperative residual dysphagia, due to the incomplete gastric myotomy and not to esophageal pouring. The aim of this study is to experimentally determine the modifications induced by Heller's myotomy and myectomy of the esophago-gastric junction on lower esophageal sphincter (LES) pressure profile, using a computerized manometric system. Myotomy of the esophageal portion of the LES (i.e. without dissection of the gastric fibers) has not modified the parameters considered, while the dissection of gastric fibers for at least 2-3 cm on the anterior gastric wall has created a significant modification of the LES pressure profile. Our observations seem to confirm and more clearly demonstrate the important role played by gastric fibers in sustaining the sphincteric HPZ. Moreover, analysis of our data, showed the need to always perform a complete myotomy. This was objectively shown during the intervention by means of intraoperative manometry, in order to significantly reduce the possibility of a dysphagic relapse, caused by inadequate treatment. PMID- 16045578 TI - Laparoscopic mesh repair antireflux surgery for treatment of large hiatal hernia. AB - SUMMARY: One of the most frequently occurring anatomic failures after laparoscopic fundoplication is migration of the wrap into the chest, with or without disruption. This so-called 'slipped' Nissen fundoplication may be the result of inadequate closure of the diaphragmatic crura or rupture of the sutures or disruption of the muscle fibers approached. From January 2000 to December 2002, a total of seven patients (four male) with a mean age of 56 years (range 22 72 years), were considered for laparoscopic antireflux procedure using DACRON mash to reinforce the crural hiatal closure. The patients were operated under general anesthesia; laparoscopy was performed by classical approach with five trocars. The mean operative time was 120 minutes (range 40-240 min). There were no deaths. The average of postoperative hospital stay was 3.5 days (range, 3-5). Patients returned to normal activities usually on postoperative day 10 (range, 7 15). The follow-up time was at least 2 years. There was only one late complication related to the use of DACRON mesh at the hiatus, due to migration of the mesh into the esophageal lumen causing disphagia. In conclusion the mesh repair antireflux surgery is a good alternative for closing the diaphragmatic defect in large hiatal hernias or to correct this problem in case of recurrence or Barrett's esophagus. PMID- 16045579 TI - Analysis of protein expression patterns in Barrett's esophagus using MALDI mass spectrometry, in search of malignancy biomarkers. AB - SUMMARY: In order to detect early changes of malignant degeneration in Barrett's esophagus (BE), and to reduce the cost of surveillance, molecular biomarkers of early malignancy have been sought, with limited success, using genomic and immunohistochemical tools. We postulate that direct analysis of epithelial proteins using mass spectrometry will provide protein profiles capable of identifying patients at high risk of developing malignancy. Our aim is to find transitional protein signals that show a cancer profile within histologically benign BE, which can be used as indicators of early malignant change. Fourteen fresh-frozen, resected esophageal cancer specimens were analyzed using laser capture microdissection and matrix-assisted laser desorption/ionization mass spectrometry. Samples of squamous epithelium, and both benign and malignant Barrett's epithelium, were compared for differences in protein expression. Reliable differentiation of squamous and Barrett's epithelium was demonstrated. A comparison of benign and malignant Barrett's epithelium identified a number of cancer-specific protein peaks that were deletion or expression variations from benign epithelium. In four instances the proteins (7350, 8446, 10850, and 14693) appeared to be early malignant changes in histologically benign BE. Mass spectrometry performed upon fresh-frozen Barrett's epithelium, obtained by laser capture microdissection, displays reproducible, tissue-specific, protein profiles. Distinct differences are demonstrated between benign and malignant epithelium, some of which appear to be candidate biomarkers of early malignant change. This technique reliably displays cellular protein expression in esophageal epithelium and deserves further study as a tool to identify early malignant degeneration in BE. PMID- 16045580 TI - Association of methylenetetrahydrofolate reductase and thymidylate synthase promoter polymorphisms with genetic susceptibility to esophageal and cardia cancer in a Chinese high-risk population. AB - SUMMARY: Methylenetetrahydrofolate reductase (MTHFR) and thymidylate synthase (TS) are key enzymes in folate metabolism, which is essential for normal DNA methylation and synthesis. Common polymorphisms at the MTHFR nucleotides position 677 (C-T) and a 28-bp tandem repeat polymorphism (2R or 3R) in the TS promoter enhancer region (TSER) have been reported to be functional and are supposed to disturb the normal DNA methylation and synthesis leading to carcinogenesis. To investigate the association between these polymorphisms and the risk of esophageal squamous cell carcinoma (ESCC) and gastric cardia adenocarcinoma (GCA), we conducted a case-control study in the Anyang area where the incidence of ESCC is highest in northern China. Subjects consisted of 275 cases with ESCC, 129 cases with GCA and 310 sex- and age-matched cancer-free controls. The risk was evaluated in terms of age-sex adjusted odds ratios (ORs) and 95% confidence intervals (CIs) by unconditional logistic regression model. The ORs for the MTHFR677TT genotype compared with the MTHFR677CC/CT genotype were 1.62 (95% CI = 1.15-2.30) and 1.81 (1.17-2.81) for ESCC and GCA, respectively. The ORs for the TSER 2R/2R genotype relative to the other genotypes were 2.44 (0.89-6.73) and 3.94 (1.29-12.0) for SCC and GCA, respectively. These findings indicated that the folate metabolism plays an important role in carcinogenesis of ESCC and GCA and the common functionally polymorphisms MTHFRC677T and TSER have substantial influence in this metabolic pathway. PMID- 16045581 TI - Overexpression of glucose transporter 1 in esophageal squamous cell carcinomas: a marker for poor prognosis. AB - SUMMARY: Recently, Glut1 (human erythrocyte glucose transporter) expression has been demonstrated in various tumors. The aim of this study is to evaluate the prognostic utility of Glut1 expression in esophageal carcinomas. We studied Glut1 expression by immunohistochemistry of paraffin sections from 63 esophageal squamous cell carcinomas. All 63 carcinomas expressed Glut1. The mean percentage of positively stained tumor cells was 77.8% (median, 84.7%). There were two staining patterns in positive cells: 'strongly positive' and 'weakly positive'. The percentage of 'strongly positive' cells (%Glut1-SP) ranged from 0% to 95.6% (mean, 32.3%; median, 27.4%). The 5-year survival rate for patients with a high %Glut1-SP (> 30%) was significantly lower than that for patients with a low %Glut1-SP (< 30%) (P < 0.01). Statistical analysis revealed that the relative risk of death for patients with high %Glut1-SP was 2.02 times that for patients with low %Glut1-SP (P = 0.064), suggesting a possible independent predictive value for %Glut1-SP. PMID- 16045582 TI - Effect of one-step 100% ethanol fixation and modified manual microdissection on high-quality RNA recovery from esophageal carcinoma specimen. AB - SUMMARY: This paper attempts to determine an optimal fixation protocol for stabilizing RNA during microdissection so as to obtain high-quality RNA from specific cell populations procured from esophageal carcinoma specimens, and to develop a manual microdissection that can facilitate the procurement. The special features of our protocol include one-step dehydration of tissue sections in 100% ethanol immediately after cryosectioning, a self-made T-shape plate (T plate) and "exclusion microdissection" procedure. The quality of RNA isolated from dissected cells was analyzed by neutral agarose gel electrophoresis and reverse transcription-polymerase chain reaction (RT-PCR) to detect genes of different abundance levels. One-step 100% ethanol fixation of cryosections effectively stabilized RNA integrity for agelong period of time while maintaining histological morphology comparable to that using the conventional procedure, indicating that it is a valid protocol for preservation of RNA in microdissected samples. In conjunction with the application of the T plate and 'exclusion microdissection' procedure, which efficiently simplifies manual microdissection procedure, allowing maximal procurement of target cells from complex primary tissues, full use of every single specimen for maximal procurement of target cells from the sections was allowed. The RNA isolated from 5 different stage specific cell populations of an esophageal carcinoma specimen was of high quality and sufficient in quantity for various downstream molecular analyses. Our method is suitable for a wide spectrum of expression analysis in diverse clinical settings. PMID- 16045583 TI - Heartburn and multiple-site foregut perforations as primary manifestation of Crohn's disease. AB - SUMMARY: Crohn's disease may affect any segment of the digestive tract, more commonly the distal ileum, colon and/or perianal region. There is an increasing number of reports dealing with foregut Crohn's disease. We present the case of a patient with a history of heartburn and multiple spontaneous perforations of the esophagus, duodenum and jejunum as a primary manifestation of Crohn's disease who required emergency surgical and endoscopic procedures. Early detection of Crohn's disease may decrease the incidence of acute life-threatening complications provided that appropriate medical treatment is administered and a multidisciplinary approach is offered to these patients. PMID- 16045584 TI - Presternal gastric bypass for late postpneumonectomy esophagopleural fistula. AB - SUMMARY: A 71-year-old diabetic patient underwent right pneumonectomy with wide mediastinal lymph node dissection for lung cancer (right upper lobe). Postoperatively he developed pleura empyema that was successfully treated - drainage and Eloesser window, followed by adjuvant radiotherapy. Two months later he developed an esophagopleural fistula. Due to the patient's physical condition primary repair of the esophageal rupture was considered a high-risk operation. Stenting was also considered as inappropriate due to the existing contamination. Bypassing with the use of the stomach as conduit was preferred due to its simplicity compared to the colon. In order to avoid mediastinum after the postradiation alterations and because of the Eloesser window we adopted a presternal subcutaneous position. Twenty-eight months after the by pass procedure the patient is in good health being able to eat and drink, has gained weight and shows no evidence of malignancy. Presternal gastric esophageal bypass has never been reported as a treatment for esophagopleural fistula. This case report indicates its possible successful use in this debilitating setting, although more experience is needed. PMID- 16045585 TI - Delayed esophageal reconstruction for aortoesophageal fistula caused by an aortic arch aneurysm with microvascular anastomosis of the left gastric artery and vein. AB - SUMMARY: We report a case of aorto esophageal fistula (AEF) with delayed esophageal reconstruction employing microvascular anastomosis. We demonstrate here that our method is useful for delayed esophageal reconstruction following AEF. PMID- 16045586 TI - Synchronous spontaneous perforation of the esophagus and a duodenal ulcer. AB - SUMMARY: Although 41% of patients with spontaneous rupture of the esophagus also suffer from gastro duodenal ulcer disease, cases of synchronous spontaneous esophageal and duodenal ulcer perforation have thus far not been reported in the literature. We report on the case of a 61-year-old man who presented with a 72 hour history of esophageal rupture and duodenal ulcer perforation. Following appropriate circulatory resuscitation we performed double resection; involving the esophagus, cardia and the distal part of the stomach, followed by substitution by means of gastro-jejunal transposition as a one-stage procedure. With reference to this case with a favorable outcome, we are presenting an analysis of indications for resectional surgery in advanced spontaneous esophageal perforation. PMID- 16045587 TI - Specialized advanced clinical placements for nurse practitioner students. PMID- 16045588 TI - Speaking with one voice. PMID- 16045589 TI - Management of seasonal allergic rhinitis: comparative efficacy of the newer generation prescription antihistamines. AB - PURPOSE: To discuss the efficacy and safety of second-generation prescription antihistamines, cetirizine, desloratadine, and fexofenadine, and to inform nurse practitioners (NP) who are in a position to optimally manage patients presenting with seasonal allergic rhinitis (SAR). DATA SOURCES: Selected published clinical trials. CONCLUSIONS: NP, who are ideally placed to successfully manage the majority of patients with SAR, should be fully aware of the treatment options available. Although first-generation over-the-counter antihistamines are convenient to use and are readily accessible, patients run the risk of experiencing adverse effects, such as sedation. For this reason, second generation antihistamines should be considered a better alternative; however, they need to be evaluated individually on the basis of their proven efficacy and safety. Both cetirizine and fexofenadine are equally effective at relieving the symptoms of SAR; however, cetirizine has been shown to be more likely to cause sedation and impairment. IMPLICATIONS FOR PRACTICE: If left untreated, SAR can have a substantial effect on patients' quality of life, resulting in fatigue, irritability, sleep disorders, and cognitive impairment. PMID- 16045590 TI - Asperger syndrome in children. AB - PURPOSE: To review Asperger syndrome characteristics, assessment tools, interventions, outcomes, and the role of the nurse practitioner in diagnosing and caring for children with Asperger syndrome. DATA SOURCES: Review of published literature on and diagnostic criteria of the condition. CONCLUSIONS: Asperger syndrome is a pervasive developmental disorder or an autism spectrum disorder that is thought to have an incidence higher than that of autism. Asperger syndrome is different from autism, with a lack of delayed language as the most distinct difference between Asperger syndrome and autism. IMPLICATIONS FOR PRACTICE: Because of the importance of early diagnosis of Asperger syndrome for outcome improvement, screening at all well-child visits from infancy on is of utmost importance to primary care pediatric nurse practitioners. With early diagnosis, timely intervention is possible, which is proven to show improvement in outcomes. PMID- 16045591 TI - Optimizing the diagnosis and treatment of migraine. AB - PURPOSE: To describe and discuss short headache questionnaires, which can simplify and improve the diagnosis of migraine. DATA SOURCES: Review of the worldwide scientific literature on short diagnostic questionnaires for migraine. CONCLUSIONS: A new three-question Headache Screen addressing disability due to recurring headaches, headache duration, and changes in headache characteristics and/or pattern over the previous 6 months displayed high sensitivity when used to survey >3000 migraineurs, correctly identifying 77% of migraineurs diagnosed by International Headache Society (IHS) criteria, clinical impression, or the presence of recurring, disabling headaches. IMPLICATIONS FOR PRACTICE: The underdiagnosis and undertreatment of migraine are problems that may be attributed to many causes involving both patients and medical providers. These include stringent diagnostic criteria established by the IHS, which fail to easily classify many common migraine presentations, the lack of clear outcome measures of successful management of migraine, and failure to recognize the iatrogenic role of prescription and nonprescription medications as an etiologic factor in chronic daily headache. The recent development of reliable, clinically useful, short headache questionnaires that are focused on headache impact facilitates the understanding and diagnosis of migraine for both patients and healthcare professionals. As a diagnostic tool, the Headache Screen has the potential to expand appropriate medication use, leading to improved functional status and quality of life for migraineurs. PMID- 16045592 TI - Comparing perceptions of cancer fatalism among African American patients and their providers. AB - PURPOSE: To describe perceptions of cancer fatalism and identify demographic correlates; to explore whether providers believe their patients are fatalistic about cancer and compare these views to the patients' views. DATA SOURCES: Both patients (n= 52) and providers (n= 35) were recruited at federally funded, community primary care centers. Data were collected using the Powe Fatalism Inventory, the Perceived Patient Fatalism Inventory, and a demographic data questionnaire. Data were analyzed using descriptive statistics, Pearson correlations, and t-test. CONCLUSIONS: The majority of patients were African American women. The majority of providers were physicians and nurses. Patients indicated low perceptions of cancer fatalism, but providers believed patients were highly fatalistic. As the patients' educational level increased, perceptions of cancer fatalism decreased. IMPLICATIONS FOR PRACTICE: The providers' belief that patients are fatalistic about cancer may influence patient-provider communication. They may be less likely to recommend screening, and patients may be less likely to initiate a discussion about cancer. Strategies are needed that target providers and their patients to address actual and/or perceived perceptions and their influence on cancer screening. PMID- 16045593 TI - The collaborative healthcare team: tensive issues warranting ongoing consideration. AB - PURPOSE: To provide a qualitative assessment of those tensive issues that warrant ongoing consideration by physicians (MDs) and advanced practice nurses (APNs) currently working in professional pairings on an interdisciplinary healthcare team. DATA SOURCES: Verbatim transcripts of semistructured interviews from five MDs and eight APNs. Questions related to what MDs and APNs should expect from others in the practice; what characteristics were valued in the colleagues with whom they were paired; and what improvements might enhance their association. CONCLUSIONS: Tensive issues were identified in five categories including autonomy and interdependence, professional role expectations, flexible role enactment, proactive problem solving, and action learning. IMPLICATIONS FOR PRACTICE: By understanding the tensive issues that could potentially influence the quality of collaboration, MDs and APNs can consider communication measures geared toward strengthening their partnerships. PMID- 16045594 TI - Calculating insulin resistance in the primary care setting: why should we worry about insulin levels in euglycemic patients? AB - PURPOSE: To describe measures that would determine which patients are insulin resistant and at risk for the metabolic syndrome and its sequelae cardiovascular diseases (CVD) and to analyze methods to determine the presence of insulin resistance and the advantages or disadvantages of each. DATA SOURCES: Review of the multidisciplinary clinical and research literature. CONCLUSIONS: Insulin resistance occurs early in the trajectory of the metabolic syndrome, making it a prime candidate for timely interventions to reduce risk for both type 2 diabetes and CVD. Therefore, prompt recognition of insulin resistance prior to the development of the full metabolic syndrome, type 2 diabetes, and/or CVD may assist in the prevention of morbidity and premature mortality. Likewise, because many insulin-resistant patients belong to minority racial groups (i.e., African American, Hispanic, Native American, or Pacific Islanders), early identification may have a positive impact on the reduction of cardiovascular health disparities. IMPLICATIONS FOR PRACTICE: Documenting the presence of insulin resistance will assist the practitioner to determine if a low-risk patient is in jeopardy for development of type 2 diabetes and/or CVD. Early cardiovascular risk identification is important to clinical practice as it allows more time for the practitioner to counsel patients for the essential planning needed to make lifestyle changes. PMID- 16045595 TI - Gastric cancer awareness campaign in Asia. PMID- 16045596 TI - Gastric cancer: Worldwide burden and prevention opportunities. AB - Gastric cancer is a major worldwide problem in developing and developed countries. It contributes substantially to the economic medical resource burden and population mortality in many countries. The absolute numbers of new cases of death are expected to increase significantly over the next decades because of the enlarging and aging of the world population. The putative factors are better understood today, and control of these factors, together with screening can help prevent this cancer from becoming an increasing burden to society. PMID- 16045597 TI - Gastritis and gastric cancer: which morphological type of Helicobacter gastritis is a precancerous risk? PMID- 16045598 TI - Etiology and prevention of gastric cancer in Linqu County, Shandong Province, China. PMID- 16045599 TI - The changing trends of the incidence of gastric cancer after Helicobacter pylori eradication in Shandong area. PMID- 16045600 TI - Eradication of Helicobacter pylori and gastric cancer: experience in Fujian Province of China. PMID- 16045601 TI - Endoscopic submucosal dissection for early gastric cancer. AB - Endoscopic mucosal resection (EMR) has been accepted as a treatment of early gastric cancer (EGC). The number of EMR procedures for EGC has been increasing because a patient's quality of life after EMR is superior to that after surgical gastrectomy. In Japan, most EMR have been performed using the strip biopsy method; however, in some cases we experienced local recurrences when multiple fragments had been resected by this method. Single-fragment resection is preferable for correct histological diagnosis and a patient's prognosis. Recently, dramatic changes have occurred in the operational technique and design of the accessory apparatus. In 1996, we developed a special endoscopic knife, called the insulation-tipped electrosurgical knife (IT knife), which consists of a conventional diathermic needle knife with a ceramic ball at the top to minimize the risk of perforation. This knife can cut submucosa safely and remove a lesion completely. We named this method endoscopic submucosal dissection (ESD). Using this procedure, 96% of the tumors (471/488) were completely resected in one piece. During the follow-up period, there were no recurrent cases. One-piece resection by ESD using the IT knife improves the quality of life and prognosis for patients with early gastric cancer. PMID- 16045602 TI - Irritable bowel syndrome and inflammatory bowel disease: interrelated diseases? AB - In the past inflammatory bowel disease (IBD), celiac disease and irritable bowel syndrome (IBS) were regarded as completely separate disorders. Now, with the description of inflammation, albeit low-grade, in IBS, and of symptom overlap between IBS and celiac disease, this contention has come under question. Is there true overlap between these disorders? Despite the limitations of available data one cannot but be struck by some areas of apparent convergence: IBD and celiac disease in remission, lymphocytic colitis and microscopic inflammation in IBS, in general, and, especially, in the post-infectious IBS category. The convergence between latent celiac disease and sub-clinical IBD, on the one hand, and IBS, on the other, appears, based on available evidence, to be somewhat spurious and may largely relate to misdiagnosis, a phenomenon which may also explain the apparent evolution of IBS into IBD in some studies. Similarities between IBS and lymphocytic colitis are more striking and less readily dismissed; as for IBS, well documented instances of progression of lymphocytic colitis to full-blown IBD are infrequent, suggesting a true separation between this disorder and classical IBD. Do IBS and lymphocytic colitis represent different responses to similar triggers? Will some of the 'inflamed' IBS subgroup be reclassified as part of the spectrum of lymphocytic colitis in the future? Will inflammation emerge as a common underlying factor in the pathogenesis of IBS? The answer to these and many questions must await further study of this fascinating area. PMID- 16045603 TI - Follow-up on the gamma-glutamyl transpeptidase in patients with severe dysplasia in an area of high incidence of esophageal cancer. AB - OBJECTIVE: To investigate the clinical value of cytological gamma-glutamyl transpeptidase (gamma-GT) staining in the reduction of the morbidity and mortality of esophageal cancer in an area of China that has a high incidence of esophageal cancer. METHODS: The results of pathology, cytological examination with gamma-GT staining in 451 patients with severe dysplasia were analyzed and followed-up for 3 years. RESULTS: At the 1-year follow-up, 17 of 231 patients with positive gamma-GT staining (7.4%) had developed esophageal cancer, but no cancer was detected in 95 patients with negative gamma-GT staining. Among 215 patients who were followed-up for 3 years, the risk of developing cancer in gamma GT staining positive patients with grade I or II severe dysplasia was 5.7% and 14.3%, respectively. However, esophageal cancer was not found in patients with negative gamma-GT staining or in patients with positive gamma-GT staining who were younger than 40 years. CONCLUSIONS: These findings suggest that gamma-GT staining can be used as a cancer risk marker of esophageal epithelial dysplasia. Esophageal dysplasia needs to be further categorized according to gamma-GT staining for esophageal cancer prevention, and intervention is required for gamma GT staining positive patients with severe dysplasia in order to increase the cost benefit ratio of esophageal cancer prevention. PMID- 16045604 TI - CD14 expression on Kupffer cells during the course of carbon tetrachloride mediated liver injury. AB - OBJECTIVE: To investigate the expression of CD14 on Kupffer cells during the course of carbon tetrachloride (CCl(4))-mediated liver injury and its role in the activation of Kupffer cells. METHODS: Rats were administered CCl(4) twice weekly for up to 8 weeks. Kupffer cells were isolated from normal and CCl(4)-treated rats by the combined 'collagenase-pronase' perfusion method, discontinuous density gradient centrifugation. On the day after isolation, the cells were incubated with RPMI-1640 containing varying doses of lipopolysaccharide (LPS) for 6 h. Supernatants were then collected for measuring the concentration of tumor necrosis factor-alpha (TNF-alpha) by enzyme-linked immunosorbent assay (ELISA). The expression of CD14 mRNA on Kupffer cells were determined by RT-PCR. The plasma concentrations of endotoxin were determined by chromogenic substrate Limulus amebocyte lysate assay. RESULTS: Basic TNF-alpha production of Kupffer cells isolated from CCl(4)-treated rats at 4 and 6 weeks was significantly higher than that of normal (P < 0.05). Following LPS stimulation the production of TNF alpha was markedly increased in Kupffer cells from the 2-, 4- and 6-week treatment groups (P < 0.05). Moreover, LPS-induced TNF-alpha production was dose dependent. CD14 mRNA expression on Kupffer cells isolated from CCl(4)-treated rats was elevated following 2 weeks of CCl(4) administration and the maximum elevation occurred at 6 weeks. Gene expression was decreased in Kupffer cells after 8 weeks of CCl(4) treatment. CCl(4) administration elicited extensive changes in liver morphology, including steatosis, inflammation and necrosis. The plasma concentrations of endotoxin of CCl(4)-treated rats were increased during the time of liver injury. CONCLUSION: Up-regulation of CD14 expression in Kupffer cells during CCl(4)-mediated chronic liver injury indicates cell activation and that they are more sensitive to LPS stimulation. Kupffer cells are critical effector cells in the early stage of liver injury. PMID- 16045605 TI - Evaluation of the inhibitory effect of antisense oligodeoxynucleotides on the growth of hepatitis C-associated hepatocellular carcinoma cells in vitro. AB - OBJECTIVE: Hepatitis C virus (HCV) RNA is invariably detected in the serum and tumor tissue of anti-HCV-positive patients with hepatocellular carcinoma (HCC). The inflammation and cirrhosis caused by HCV could be the promoter for development of HCC or HCC could be the consequence of HCV infection independent of the effect of cirrhosis. The ability of the core protein of HCV to modulate gene transcription, cell proliferation and cell death by interacting with cellular genes that regulate cell growth and differentiation is involved in the pathogenesis of HCC. HCV NS3 protease is an attractive target for antiviral agent development because it is required for viral replication. Recent studies that constructed an in vitro model of HCC demonstrated that antisense oligodeoxynucleotides (AS-ODN) interfered with NS3 translation in a dose dependent fashion and significantly inhibited protease activity. We studied the in vitro effect of AS-ODN on the rate of growth of the HCC cells grown in culture associated with HCV. METHODS: Core biopsy was taken from 20 patients with HCC associated with HCV and each one was divided into two parts: group I to which antisense was added and group II which served as a control group. Comparison of cell viability between tubes with and without AS-ODN was done using MTT assay, LDH assay, cell cycle analysis, trypan blue exclusion test and colony formation in soft agar. RESULTS: Colony formation in soft agar was inhibited in group I compared with the control group and the inhibition was highly significant (P < 0.01). The LDH concentration in culture supernatant and the trypan blue exclusion test, both reflecting cellular death, was higher in group I than group II and the difference was highly significant (P < 0.01). MTT assay showed a highly significant decrease in cell activation in group I than in group II (P < 0.01). The percentage of cells in the G(0)/G(1) phase was higher in group I than in group II and the difference was significant (P = 0.04). There was an insignificant difference between both groups in the percentage of cells in S phase (P = 0.378). The inhibitory effect of AS-ODNs on tumor cells in G(2)/M phase was highly significant compared with the control group (P < 0.01). CONCLUSIONS: AS-ODN has a significant inhibitory effect on the growth of HCV associated HCC cells grown in fluid culture, and there is potential for the use of AS-ODN as oncotherapy. PMID- 16045606 TI - Fighting an enemy within: cytoplasmic inhibitors of bacterial cysteine proteases. AB - The genes encoding secreted, broad-spectrum activity cysteine proteases of Staphylococcus spp. (staphopains) and Streptococcus pyogenes (streptopain, SpeB) are genetically linked to genes encoding cytoplasmic inhibitors. While staphopain inhibitors have lipocalin-like folds, streptopain is inhibited by a protein bearing the scaffold of the enzyme profragment. Bioinformatic analysis of other prokaryotic genomes has revealed that two more species may utilize this same genetic arrangement to control streptopain-like proteases with lipocalin-like inhibitors, while three other species may employ a C-terminally located domain that resembles the profragment. This apparently represents a novel system that bacteria use to control the intracellular activity of their proteases. PMID- 16045607 TI - Where asymmetry in gene expression originates. AB - A general problem in developmental biology concerns the process by which cells of one type divide to give dissimilar daughter cells. Even though these daughter cells may be genetically identical, they can differ morphologically and physiologically and have different fates. As one of the simplest differentiation processes, Bacillus subtilis sporulation represents an excellent model system for studying cell differentiation. Several decades of study of this process have provided insight into cell cycle regulation and development. This review summarizes important advances in our understanding of asymmetric gene expression during spore formation with an emphasis on developmental stages that lead to asymmetric septum formation and especially to activation of the first compartment specific sigma factor -sigma(F). PMID- 16045608 TI - The phage-shock-protein response. AB - The phage-shock-protein (Psp) system responds to extracytoplasmic stress that may reduce the energy status of the cell. It is conserved in many different bacteria and has been linked to several important phenotypes. Escherichia coli psp mutants have defects in maintenance of the proton-motive force, protein export by the sec and tat pathways, survival in stationary phase at alkaline pH, and biofilm formation. Yersinia enterocolitica psp mutants cannot grow when the secretin component of a type III secretion system is mislocalized, and have a severe virulence defect in animals. A Salmonella enterica psp mutation exacerbates some phenotypes of an rpoE null mutant and the psp genes of S. enterica and Shigella flexneri are highly induced during macrophage infection. PspA, the most abundant of the Psp proteins, is required for most of the phenotypes associated with the Psp system. Therefore, PspA is probably an effector that may play a role in maintaining cytoplasmic membrane integrity and/or the proton-motive force. However, PspA is not required for the ability to tolerate secretin mislocalization, which suggests an important physiological role for other Psp proteins. This article summarizes our current understanding of the Psp system: inducing signals, the underlying signal transduction mechanisms, the physiological roles it may play, and a genomic analysis of its conservation. PMID- 16045609 TI - C-di-GMP: the dawning of a novel bacterial signalling system. AB - Bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) has come to the limelight as a result of the recent advances in microbial genomics and increased interest in multicellular microbial behaviour. Known for more than 15 years as an activator of cellulose synthase in Gluconacetobacter xylinus, c-di-GMP is emerging as a novel global second messenger in bacteria. The GGDEF and EAL domain proteins involved in c-di-GMP synthesis and degradation, respectively, are (almost) ubiquitous in bacterial genomes. These proteins affect cell differentiation and multicellular behaviour as well as interactions between the microorganisms and their eukaryotic hosts and other phenotypes. While the role of GGDEF and EAL domain proteins in bacterial physiology and behaviour has gained appreciation, and significant progress has been achieved in understanding the enzymology of c-di-GMP turnover, many questions regarding c-di-GMP-dependent signalling remain unanswered. Among these, the key questions are the identity of targets of c-di-GMP action and mechanisms of c-di-GMP-dependent regulation. This review discusses phylogenetic distribution of the c-di-GMP signalling pathway in bacteria, recent developments in biochemical and structural characterization of proteins involved in its metabolism, and biological processes affected by c-di GMP. The accumulated data clearly indicate that a novel ubiquitous signalling system in bacteria has been discovered. PMID- 16045610 TI - Specificity and selectivity determinants of peptide transport in Lactococcus lactis and other microorganisms. AB - Peptide transport in microorganisms is important for nutrition of the cell and various signalling processes including regulation of gene expression, sporulation, chemotaxis, competence and virulence development. Peptide transport is mediated via different combinations of ion-linked and ATP-binding cassette (ABC) transporters, the latter utilizing single or multiple peptide-binding proteins with overlapping specificities. The paradigm for research on peptide transport is Lactococcus lactis, in which the uptake of peptides containing essential amino acids is vital for growth on milk proteins. Differential expression and characteristics of peptide-binding proteins in several Lactococcus lactis strains resulted in apparent conflicts with older literature. Recent developments and new data now make the pieces of the puzzle fall back into place again and confirm the view that the oligopeptide-binding proteins determine the uptake selectivity of their cognate ABC transporters. Besides reviewing the current data on binding specificity and transport selectivity of peptide transporters in L. lactis, the possible implications for peptide utilization by other bacterial species are discussed. PMID- 16045611 TI - SpeB-Spi: a novel protease-inhibitor pair from Streptococcus pyogenes. AB - This study presents evidence for a novel protease-protease inhibitor couple, SpeB Spi, in the human pathogen Streptococcus pyogenes. The gene for the inhibitor Spi is located directly downstream of the gene for the streptococcal cysteine protease SpeB. Spi is 37% identical and 70% similar to the sequence of the SpeB propeptide, suggesting that Spi and the SpeB propeptide might bind to SpeB in an analogous manner. Secondary structure predictions and molecular modelling suggested that Spi would adopt a structure similar to the SpeB propeptide. The spi gene was co-transcribed with speB on the 1.7 knt and 2.2 knt transcripts previously identified for speB. The Spi protein was purified by SpeB-affinity chromatography from the S. pyogenes cytoplasm. Recombinant Spi was produced and purified, and shown to bind to SpeB and to inhibit its protease activity. Although a similar genetic arrangement of protease and inhibitor is present in staphylococci, this is the first example of an inhibitor molecule that is a structural homologue of the cognate propeptide, and which is genetically linked to the protease gene. Thus, this represents a novel system whereby bacteria may control the intracellular activity of their proteases. PMID- 16045612 TI - Interaction of Rv1625c, a mycobacterial class IIIa adenylyl cyclase, with a mammalian congener. AB - The adenylyl cyclase Rv1625c from Mycobacterium tuberculosis codes for a protein with six transmembrane spans and a catalytic domain, i.e. it corresponds to one half of the pseudoheterodimeric mammalian adenylyl cyclases (ACs). Rv1625c is active as a homodimer. We investigated the role of the Rv1625c membrane domain and demonstrate that it efficiently dimerizes the protein resulting in a 7.5-fold drop in K(m) for ATP. Next, we generated a duplicated Rv1625c AC dimer by a head to-tail concatenation. This produced an AC with a domain order exactly as the mammalian pseudoheterodimers. It displayed positive cooperativity and a 60% increase of v(max) compared with the Rv1625c monomer. Further, we probed the compatibility of mycobacterial and mammalian membrane domains. The second membrane anchor in the Rv1625c concatamer was replaced with membrane domain I or II of rabbit type V AC. The mycobacterial and either mammalian membrane domains are compatible with each other and both recombinant proteins are active. A M. tuberculosis Rv1625c knockout strain was assayed in a mouse infection model. In vitro growth characteristics and in vivo organ infection and mortality were unaltered in the knockout strain indicating that AC Rv1625c alone is not a virulence factor. PMID- 16045613 TI - DNA polymerase I acts in translesion synthesis mediated by the Y-polymerases in Bacillus subtilis. AB - Translesion synthesis (TLS) across damaged DNA bases is most often carried out by the ubiquitous error-prone DNA polymerases of the Y-family. Bacillus subtilis encodes two Y-polymerases, Pol Y1 and Pol Y2, that mediate TLS resulting in spontaneous and ultraviolet light (UV)-induced mutagenesis respectively. Here we show that TLS is a bipartite dual polymerase process in B. subtilis, involving not only the Y-polymerases but also the A-family polymerase, DNA polymerase I (Pol I). Both the spontaneous and the UV-induced mutagenesis are abolished in Pol I mutants affected solely in the polymerase catalytic site. Physical interactions between Pol I and either of the Pol Y polymerases, as well as formation of a ternary complex between Pol Y1, Pol I and the beta-clamp, were detected by yeast two- and three-hybrid assays, supporting the model of a functional coupling between the A- and Y-family polymerases in TLS. We suggest that the Pol Y carries the synthesis across the lesion, and Pol I takes over to extend the synthesis until the functional replisome resumes replication. This key role of Pol I in TLS uncovers a new function of the A-family DNA polymerases. PMID- 16045614 TI - HilD, HilC and RtsA constitute a feed forward loop that controls expression of the SPI1 type three secretion system regulator hilA in Salmonella enterica serovar Typhimurium. AB - Salmonella enterica serovar Typhimurium invades intestinal epithelial cells using a type three secretion system (TTSS) encoded on Salmonella Pathogenicity Island 1 (SPI1). The SPI1 TTSS injects effector proteins into the cytosol of host cells where they promote actin rearrangement and engulfment of the bacteria. We previously identified RtsA, an AraC-like protein similar to the known HilC and HilD regulatory proteins. Like HilC and HilD, RtsA activates expression of SPI1 genes by binding upstream of the master regulatory gene hilA to induce its expression. HilA activates the SPI1 TTSS structural genes. Here we present evidence that hilA expression, and hence the SPI1 TTSS, is controlled by a feedforward regulatory loop. We demonstrate that HilC, HilD and RtsA are each capable of independently inducing expression of the hilC, hilD and rtsA genes, and that each can independently activate hilA. Using competition assays in vivo, we show that each of the hilA regulators contribute to SPI1 induction in the intestine. Of the three, HilD has a predominant role, but apparently does not act alone either in vivo or in vitro to sufficiently activate SPI1. The two-component regulatory systems, SirA/BarA and OmpR/EnvZ, function through HilD, thus inducing hilC, rtsA and hilA. However, the two-component systems are not responsible for environmental regulation of SPI1. Rather, we show that 'SPI1 inducing conditions' cause independent activation of the rtsA, hilC and hilD genes in the absence of known regulators. Our model of SPI1 regulation provides a framework for future studies aimed at understanding this complicated regulatory network. PMID- 16045615 TI - Disruption of the developmental programme of Trypanosoma brucei by genetic ablation of TbZFP1, a differentiation-enriched CCCH protein. AB - The regulation of differentiation is particularly important in microbial eukaryotes that inhabit multiple environments. The parasite Trypanosoma brucei is an extreme example of this, requiring exquisite gene regulation during transmission from mammals to the tsetse fly vector. Unusually, trypanosomes rely almost exclusively on post-transcriptional mechanisms for regulated gene expression. Hence, RNA binding proteins are potentially of great significance in controlling stage-regulated processes. We have previously identified TbZFP1 as a trypanosome molecule transiently enriched during differentiation to tsetse midgut procyclic forms. This small protein (101 amino acids) contains the unusual CCCH zinc finger, an RNA binding motif. Here, we show that genetic ablation of TbZFP1 compromises repositioning of the mitochondrial genome, a specific event in the strictly regulated differentiation programme. Despite this, other events that occur both before and after this remain intact. Significantly, this phenotype correlates with the TbZFP1 expression profile during differentiation. This is the first genetic disruption of a developmental regulator in T. brucei. It demonstrates that programmed events in parasite development can be uncoupled at the molecular level. It also further supports the importance of CCCH proteins in key aspects of trypanosome cell function. PMID- 16045616 TI - Bacillus anthracis CapD, belonging to the gamma-glutamyltranspeptidase family, is required for the covalent anchoring of capsule to peptidoglycan. AB - Several examples of bacterial surface-structure anchoring have been described, but they do not include polyglutamate capsule. Bacillus anthracis capsule, which is composed only of poly-gamma- d-glutamate, is one of the two major virulence factors of the bacterium. We analysed its anchoring. We report that the polyglutamate is anchored directly to the peptidoglycan and that the bond is covalent. We constructed a capD mutant strain, capD being the fourth gene of the capsule biosynthetic operon. The mutant bacilli are surrounded by polyglutamate material that is not covalently anchored. Thus, CapD is required for the covalent anchoring of polyglutamate to the peptidoglycan. Sequence similarities suggest that CapD is a gamma-glutamyltranspeptidase. Furthermore, CapD is cleaved at the gamma-glutamyltranspeptidase consensus cleavage site, and the two subunits remain associated, as necessary for gamma-glutamyltranspeptidase activity. Other Gram positive gamma-glutamyltranspeptidases are secreted, but CapD is located at the Bacillus surface, associated both with the membrane and the peptidoglycan. Polyglutamate is hydrolysed by CapD indicating that it is a CapD substrate. We suggest that CapD catalyses the capsule anchoring reaction. Interestingly, the CapD(-) strain is far less virulent than the parental strain. PMID- 16045617 TI - Horizontal transfer of Yersinia high-pathogenicity island by the conjugative RP4 attB target-presenting shuttle plasmid. AB - The high-pathogenicity island (HPI) encodes a highly efficient yersiniabactin system of iron acquisition responsible for mouse lethality in Yersinia. Although the HPI is widely disseminated among Enterobacteriaceae it lacks functions necessary for its replication and transmission. Therefore, the mechanism of its horizontal transfer and circulation is completely obscure. On the other hand, the HPI is a genetically active island in the bacterial cell. It encodes a functional recombinase and is able to transpose to new targets on the chromosome. Here we report on a possible mechanism of the HPI dissemination based on site-specific recombination of the excised HPI with the attB-presenting (asn tRNA gene) RP4 promiscuous conjugative shuttle plasmid. The resulting cointegrate can be transferred by conjugation to a new host, where it dissociates, and the released HPI integrates into any unoccupied asn tRNA gene target in the genome. This mechanism has been proven both with the 'mini' island carrying only the attP recognition site and genes coding for recombination enzymes and with the complete HPI labelled with an antibiotic resistance marker. After acquisition of the mobilized complete form of the HPI, the ability of the HPI-cured Yersinia enterocolitica WA-TH(-) strain to produce yersiniabactin has been restored. Such 'trapping' of pathogenicity islands and subsequent shuffling to new hosts by a conjugative replicon carrying a suitable attB site could be applied to other functional integrative elements and explain wide dissemination of PAIs. PMID- 16045618 TI - NssR, a member of the Crp-Fnr superfamily from Campylobacter jejuni, regulates a nitrosative stress-responsive regulon that includes both a single-domain and a truncated haemoglobin. AB - Consistent with its role as a nitric oxide (NO)-detoxifying globin in Campylobacter jejuni, Cgb (Campylobacter globin) expression is strongly and specifically induced following exposure to nitrosative stress, suggesting a previously unrecognized capacity for NO-related stress sensing in this food-borne pathogen. In this study, Fur and PerR have been eliminated as major regulators of cgb, and NssR (Cj0466), a member of the Crp-Fnr superfamily, has been identified as the major positive regulatory factor that controls nitrosative stress responsive expression of this gene. Accordingly, disruption of nssR resulted in the abolition of inducible cgb expression, which was restored by a complementing chromosomal insertion of the wild-type gene with its indigenous promoter at a second location. The NssR-deficient mutant was more sensitive to NO-related stress than a cgb mutant and this phenotype most likely arises from the failure of these cells to induce other NO-responsive components in addition to Cgb. Indeed, analysis of global gene expression, by microarray and confirmatory real time polymerase chain reaction (PCR) in the wild type and nssR mutant, not only confirmed the dependence of inducible cgb expression on NssR, but also revealed for the first time a novel NssR-dependent nitrosative stress-responsive regulon. This regulon of at least four genes includes Cj0465c, a truncated globin. Consistent with NssR being a Crp-Fnr superfamily member, an Fnr-like binding sequence (TTAAC-N(4)-GTTAA) was found upstream of each gene at locations -40.5 to -42.5 relative to the centre of the binding sites and the transcription start point. Site-directed mutagenesis confirmed that this cis-acting motif mediates the nitrosative stress-inducible expression of cgb. PMID- 16045619 TI - Polyphosphate kinase regulates error-prone replication by DNA polymerase IV in Escherichia coli. AB - The ppk gene encodes polyphosphate kinase (Ppk), an enzyme that catalyses the polymerization of inorganic phosphate into long chains of polyphosphate (polyP). An insertion mutation in ppk causes a decrease in adaptive mutation in Escherichia coli strain FC40. Adaptive mutation in FC40 mostly results from error prone DNA polymerase IV (Pol IV), encoded by dinB; most of the antimutagenic phenotype of the ppk mutant disappears in a dinB mutant strain. In addition, the ppk mutant causes a decrease in growth-dependent mutations produced by overexpressing Pol IV. However, the amount of Pol IV protein is unchanged in the ppk mutant strain, indicating that the activity or fidelity of Pol IV is altered. Adaptive mutation is inhibited both by the absence of Ppk, which results in low amounts of polyP, and by overproduction of Ppk, which results in high amounts of polyP, suggesting that an optimal level of polyP is necessary. Taken together, these results suggest a novel mechanism involving polyP that directly or indirectly regulates DNA polymerase activity or fidelity. PMID- 16045620 TI - BrgE is a regulator of Myxococcus xanthus development. AB - We report here the identification and characterization of a member of the Myxococcus xanthus SdeK signal transduction pathway, BrgE. This protein was identified as an SdeK-interacting component using a yeast two-hybrid screen, and we further confirmed this interaction by the glutathione S-transferase (GST) pulldown assay. Additional yeast two-hybrid analyses revealed that BrgE preferentially interacts with the putative amino-terminal sensor domain of SdeK, but not with the carboxy-terminal kinase domain. A brgE insertion strain was shown to be blocked in development between aggregation and mound formation, and decreased by 50-fold in viable spore production compared with the parental wild type. These phenotypes are similar to those of sdeK mutants. The brgE mutation also altered expression of a sample of Tn5 lac developmental markers that are also SdeK regulated. Finally, we demonstrated that a brgE sdeK double mutant has a more severe sporulation defect than either of the two single mutants, suggesting that BrgE and SdeK act synergistically to regulate wild-type levels of sporulation. In sum, these data suggest that BrgE operates as an auxiliary factor to stimulate the SdeK signal transduction pathway by directly binding to the amino-terminal sensor domain of SdeK. PMID- 16045621 TI - Large increases in attractant concentration disrupt the polar localization of bacterial chemoreceptors. AB - In bacterial chemotaxis, the chemoreceptors [methyl-accepting chemotaxis proteins (MCPs)] transduce chemotactic signals through the two-component histidine kinase CheA. At low but not high attractant concentrations, chemotactic signals must be amplified. The MCPs are organized into a polar lattice, and this organization has been proposed to be critical for signal amplification. Although evidence in support of this model has emerged, an understanding of how signals are amplified and modulated is lacking. We probed the role of MCP localization under conditions wherein signal amplification must be inhibited. We tested whether a large increase in attractant concentration (a change that should alter receptor occupancy from c. 0% to > 95%) would elicit changes in the chemoreceptor localization. We treated Escherichia coli or Bacillus subtilis with a high level of attractant, exposed cells to the cross-linking agent paraformaldehyde and visualized chemoreceptor location with an anti-MCP antibody. A marked increase in the percentage of cells displaying a diffuse staining pattern was obtained. In contrast, no increase in diffuse MCP staining is observed when cells are treated with a repellent or a low concentration of attractant. For B. subtilis mutants that do not undergo chemotaxis, the addition of a high concentration of attractant has no effect on MCP localization. Our data suggest that interactions between chemoreceptors are decreased when signal amplification is unnecessary. PMID- 16045622 TI - MsmR, a specific positive regulator of the Streptococcus pyogenes FCT pathogenicity region and cytolysin-mediated translocation system genes. AB - As a prerequisite for colonization or causing local infections, Streptococcus pyogenes (group A streptococci, GAS) need to specifically adhere to eukaryotic cell surfaces. Predominantly responsible adhesin genes are contained in a genotype-specific pattern within the FCT region of the GAS genome. In this study, MsmR, belonging to AraC/XylS type transcriptional regulators, was identified in the FCT region as a positive regulator of the major fibronectin-binding adhesin protein F2 in a serotype M49 strain. Compared with the wild-type strain, the msmR mutant showed reduced binding to immobilized fibronectin and decreased adherence to and internalization into human pharyngeal epithelial cells. These results suggested that altered levels of fibronectin-binding proteins in the mutant affect eukaryotic cell attachment and internalization. Complete transcriptome and reporter fusion assay data revealed that MsmR positively regulates FCT region genes including Nra and cytolysin-mediated translocation system genes. Consistent with the genetic data, the mutant showed attenuated streptolysin O activity and eukaryotic cell cytotoxity. Direct binding of recombinant MsmR to nga, nra/cpa and prtF2 promoter regions was confirmed by EMSA assays. As prior analysis demonstrated the Nra regulator negatively affects gene expression from the FCT region, MsmR and Nra appear to adversely control crucial virulence factor expression in GAS and thus contribute to a fine-tuned balance between local destructive process and metastatic spreading of the bacteria. PMID- 16045623 TI - Roles for fibrinogen, immunoglobulin and complement in platelet activation promoted by Staphylococcus aureus clumping factor A. AB - Staphylococcus aureus is an important cause of infective endocarditis (IE) in patients without a history of prior heart valve damage. The ability to stimulate the activation of resting platelets and their subsequent aggregation is regarded as an important virulence factor of bacteria that cause IE. Clumping factor A is the dominant surface protein responsible for platelet activation by S. aureus cells in the stationary phase of growth. This study used Lactococcus lactis as a surrogate host to study the mechanism of ClfA-promoted platelet activation. Expression of ClfA from a nisin-inducible promoter demonstrated that a minimum level of surface-expressed ClfA was required. Using platelets that were purified from plasma, the requirement for both bound fibrinogen and immunoglobulin was demonstrated. The immunoglobulin G (IgG) requirement is consistent with the potent inhibition of platelet activation by a monoclonal antibody specific for the platelet FcgammaRIIa receptor. Furthermore the IgG must contain antibodies specific for the ClfA A domain. A model is proposed whereby bacterial cells armed with a sufficient number of surface-bound fibrinogen molecules can engage resting platelet glycoprotein GPIIb/IIIa, aided by bound IgG molecules, which encourages the clustering of FcgammaRIIa receptors. This can trigger activation of signal transduction leading to activation of GPIIb/IIIa and aggregation of platelets. In addition, analysis of a mutant of ClfA totally lacking the ability to bind fibrinogen revealed a second, although less efficient, mechanism of platelet activation. The fibrinogen-independent pathway required IgG and complement deposition to trigger platelet aggregation. PMID- 16045624 TI - Ability of IncP-9 plasmid pM3 to replicate in Escherichia coli is dependent on both rep and par functions. AB - IncP-9 plasmids are common in Pseudomonas species and can be transferred to other Gram-negative eubacteria but tend not to be stably maintained outside their natural host genus. A 1.3 kb ori V-rep fragment from IncP-9 plasmid pM3 was sufficient for autonomous replication in Pseudomonas putida but not in Escherichia coli. Replication of ori V-rep in E. coli was restored when additional rep was provided in trans, suggesting that the replication defect resulted from insufficient rep expression from its natural promoter. A promoter deficiency in E. coli was confirmed by reporter gene assays, transcriptional start point mapping and mutation of the promoter recognition elements. Dissection of the pM3 mini-replicon, pMT2, showed that this replication deficiency in E. coli is suppressed by additional determinants from its par operon: ParB, which can be supplied in trans, and its target, the par operon promoter, required in cis to ori V-rep. We propose that ParB binding to its target either changes plasmid DNA and thus promoter conformation or by spreading or looping contacts RNAP at the rep promoter so that rep expression is sufficient to activate ori V. PMID- 16045625 TI - Zap1 activation domain 1 and its role in controlling gene expression in response to cellular zinc status. AB - The Zap1 transcription factor is a central player in zinc homeostasis in yeast. This protein regulates the expression of genes involved in zinc accumulation and storage. For most of its target genes, Zap1 activates expression in zinc-limited cells and this function is inhibited in replete cells. Zap1 has two activation domains, AD1 and AD2, which are independently regulated by zinc status. In this study, we characterized AD1 and its regulation by zinc. AD1 was mapped using deletions to residues 332-402 of Zap1. The region required for the zinc responsiveness of this activation domain, designated 'ZRD(AD1), was mapped to residues 182-502. Thus, AD1 is embedded within its larger zinc-responsive domain. Using a combination of in silico analysis, random mutagenesis and site-directed mutagenesis, we identified key residues within ZRD(AD1) required for its regulation by zinc. Most of these residues are cysteines and histidines that could potentially serve as Zn(II) ligands. These results suggest that ZRD(AD1) senses zinc by direct Zn(II) binding. Consistent with this hypothesis, purified ZRD(AD1) bound multiple Zn(II) ions. Finally, our results indicate that, in the context of the full-length Zap1 protein, AD1 and AD2 are both critical to the full control of gene expression in response to zinc. PMID- 16045626 TI - Function of the thioredoxin proteins in Cryptococcus neoformans during stress or virulence and regulation by putative transcriptional modulators. AB - The thioredoxin system, consisting of thioredoxin, thioredoxin reductase and NADPH, is known to protect cells against oxidative stress. This disulphide reducing system is present in Cryptococcus neoformans and consists of two small, dithiol thioredoxin proteins and one thioredoxin reductase. In this study, we describe the thioredoxin proteins, Trx1 and Trx2, and present their importance not only to stress resistance, but also to the virulence of C. neoformans. Using real-time polymerase chain reaction, we show the induction of both thioredoxin genes during oxidative and nitrosative stress. We describe through deletion studies that the trx1delta mutant has a severe growth defect and is sensitive to multiple stresses, while the trx2delta mutant is only sensitive to nitric oxide stress. Using gene replacement studies, we demonstrate that the thioredoxin protein products are partially redundant in function, but there is differential gene regulation which is especially important to nitrosative stress resistance. We have also identified two putative transcription factors, Atf1 and Yap4, which appear to differentially regulate the thioredoxin system under different conditions. Atf1 is necessary for oxidative stress induction, and Yap4 is necessary for nitrosative stress induction of the thioredoxin genes in C. neoformans. While these two putative transcription factors each appear to be dispensable for survival in macrophages and virulence in mice, we show the more highly expressed thioredoxin, TRX1, is necessary for survival of C. neoformans in the oxidative environment of macrophages and important for virulence of this fungal pathogen. PMID- 16045627 TI - Mycobacterium tuberculosis appears to lack alpha-ketoglutarate dehydrogenase and encodes pyruvate dehydrogenase in widely separated genes. AB - Mycobacterium tuberculosis (Mtb) persists for prolonged periods in macrophages, where it must adapt to metabolic limitations and oxidative/nitrosative stress. However, little is known about Mtb's intermediary metabolism or antioxidant defences. We recently identified a peroxynitrite reductase-peroxidase complex in Mtb that included products of the genes sucB and lpd, which are annotated to encode the dihydrolipoamide succinyltransferase (E2) and lipoamide dehydrogenase (E3) components of alpha-ketoglutarate dehydrogenase (KDH). However, we could detect no KDH activity in Mtb lysates, nor could we reconstitute KDH by combining the recombinant proteins SucA (annotated as the E1 component of KDH), SucB and Lpd. We therefore renamed the sucB product dihydrolipoamide acyltransferase (DlaT). Mtb lysates contained pyruvate dehydrogenase (PDH) activity, which was lost when the dlaT gene (formerly, sucB) was disrupted. Purification of PDH from Mtb yielded AceE, annotated as an E1 component of PDH, along with DlaT and Lpd. Moreover, anti-DlaT antibody coimmunoprecipitated AceE. Finally, recombinant AceE, DlaT and Lpd, although encoded by genes that are widely separated on the chromosome, reconstituted PDH in vitro with Km values typical of bacterial PDH complexes. In sum, Mtb appears to lack KDH. Instead, DlaT and Lpd join with AceE to constitute PDH. PMID- 16045629 TI - Assessing obesity: are ethnic differences in body mass index and waist classification criteria justified? PMID- 16045630 TI - Prevalence of obesity in Canada. AB - Excess weight represents a critical and common health problem in Canada. The last survey of a national representative sample based on measured anthropometrics has been conducted in 1992. According to surveys using measured data, the prevalence of obesity (body mass index, BMI = 30.0 kg m(-2)) between 1970 and 1992 for those aged 20-69 years increased from 8% to 13% in men and 13% to 15% in women. The proportion of Canadians displaying a BMI > or =25.0 kg m(-2) increased from 47% to 58% in men and from 34% to 41% in women in the same period. The most recent prevalence estimates from self-reported data in a national representative sample indicated that 15% of the adult population (> or =18 years) was affected by obesity, while an additional 33% was classified in the overweight category (BMI 25.0-29.9 kg m(-2)) in 2003. However, it has been suggested that self-reported height and weight underestimate the prevalence of obesity by approximately 10%. Canadian children, aboriginal populations, and immigrants are some of the vulnerable groups particularly at risk of excess weight or for which the increase in the recent decades has been greater than the national increase. The increases in overweight and obesity over the past 30 years among Canadians have been dramatic. It will be possible to precisely analyse the current situation and its evolution in the last 10 years when data based on measured height and weight will be released, that is, in 2005 and after. PMID- 16045631 TI - Prevalence of obesity in Australia. AB - Australia is a large country approximately equal in area to mainland United States. The relatively small population of around 20 million are composed primarily of Caucasians. Extensive immigration from many different countries has made Australia one of the most culturally diverse populations in the world. Indigenous Australians make up only 2.4% of the total population. Australia has a prosperous Western-style capitalist economy, and spends approximately 830 million dollars on the direct health care costs of obesity. For Australians, it is now more common to have a weight problem, with overweight affecting 48% of men and 30% of women and obesity affecting a further 19% of men and 22% of women. This paper reports on recent epidemiological studies documenting the extent of overweight and obesity in adults and children in Australia. PMID- 16045632 TI - Prevalence of obesity in Iran. AB - Recent descriptive studies suggest that Iran has geared in the nutrition and epidemiological transition processes. Therefore, while the problems of undernutrition (e.g. growth retardation and micronutrient deficiencies) still exist, the burden of overweight/obesity and diet-related chronic diseases is increasing. The prevalence of overweight (body mass index > or = 85th reference percentiles) among urban 15-39 and 40-69 year olds is estimated at about 22% and 40% respectively. Corresponding values in rural areas are 16% and 26%. The transition seems faster among female population at national level. There are however, great differences between different provinces. Urgent preventive strategies are needed to simultaneously tackle both forms of malnutrition in the country. PMID- 16045633 TI - Ethnic comparisons of obesity in the Asia-Pacific region: protocol for a collaborative overview of cross-sectional studies. AB - The prevalence of overweight and obesity has been rapidly increasing within countries of the Asia-Pacific region, with adverse consequences for health. The Obesity in Asia Collaboration (OAC) was initiated to provide reliable evidence concerning the relationships between anthropometrical markers of adiposity with cardiovascular risk factors within, and across, ethnic groups. This is to be achieved through the collection of cross-sectional studies from the Asia-Pacific region. These data will be used to examine the ethnic-specific associations of different anthropometric measures of adiposity (such as body mass index, waist circumference, waist-hip ratio and waist-height ratio) with cardiovascular risk factors (fasting blood glucose and blood pressure). Estimates of the relative risks for type 2 diabetes and hypertension with increased adiposity will be obtained using data from contributing studies in conjunction with those from the previously established Asia Pacific Cohort Studies Collaboration. To date, 12 countries and regions from the Asia-Pacific region with information on nearly 300,000 individuals, are participants in the OAC. Data from other eligible studies are sought. The findings from the OAC will provide reliable evidence concerning the association of adiposity with cardiovascular risk factors across ethnic groups. It will also provide a strong evidence base upon which to define ethnic-specific cut-points for overweight and obesity. PMID- 16045634 TI - The fundamental flaw in obesity research. AB - The basic problem with comparative diet trials is our inability to measure what people eat. All conventional instruments depend on subjects' reports. Most trials lack independent biochemical, physiological or genetic measures of intake. So, we do not know if subjects actually follow the diets being tested and compared. We can assess weight gain/loss, but we fail in a fundamental scientific requirement, accurately measuring the independent variable in a causal experiment. Worse, we know most subjects under-report their energy intake and its components, the obese especially. The problem is compounded by attempts to show diets' effects on other risk factors, like triglycerides. Researchers seek to correlate two variables, without having accurately measured one of them, producing misleading associations. The consequence is we do not know if the results of any current diet trials are valid or reliable. Developing rigorous measures of food intake is the highest priority in obesity research. That involves improvements in technology as well as science. We need: (1) biomarkers of intake for energy, macro- and micro-nutrients and other food components relevant to weight gain/loss; (2) field measuring instruments that are cheap, rapid, painless, non intrusive and self-administerable; and (3) electronic data transmission systems that preclude subjects' ability to misreport. PMID- 16045635 TI - Evidence of a possible link between obesogenic food advertising and child overweight. AB - A recent review of the literature concluded that advertising of foods on television may influence children's food choices and encourage unhealthy diets, but the review acknowledged there was a lack of clear evidence in coming to this conclusion. The present paper examines ecological evidence for a link between advertising to children and the risk of overweight using data from surveys of advertising on children's television and estimates of the prevalence of overweight among children, in the USA, Australia and eight European countries. A significant association was found between the proportion of children overweight and the numbers of advertisements per hour on children's television, especially those advertisements that encourage the consumption of energy-dense, micronutrient-poor foods (r = 0.81, P < 0.005). A weaker, negative association was found between the proportion of children overweight and the number of advertisements encouraging healthier diets (r = -0.56, P < 0.10). The quantity of advertising on children's television appears to be related to the prevalence of excess body weight among children. Furthermore, the content of the advertising appears to have a specific effect. The findings justify the need for taking precautionary measures to reduce children's exposure to obesogenic marketing practices. PMID- 16045636 TI - Utility and validity of the eating behavior inventory in clinical obesity research: a review of the literature. AB - The Eating Behavior Inventory (EBI) is a questionnaire designed to assess behaviours associated with weight loss and weight management. In the 25 years since it was first published, the EBI has been used to examine weight management behaviours of adults and children in more than 20 weight loss investigations. This paper systematically reviews the use of the EBI in these studies. Results indicate that baseline EBI scores have remained quite consistent over time and are generally not influenced by patient variables. The EBI has also been shown to be consistently sensitive to behavioural weight management interventions, but it appears that the amount of change in EBI scores has decreased slightly over time. Finally, the amount of change in EBI score with treatment typically correlates positively with the amount of weight subjects lose. Overall, the research that has used the EBI has provided solid and consistent evidence of the utility of the EBI as a valid tool for assessing weight management behaviours. PMID- 16045637 TI - Laparophobia: a cognitive perspective on appetite control in anorexia nervosa. AB - Decades of research have demonstrated that anorexia nervosa (AN) may be associated with aberrant cognition, yet, its role in maintaining stringent dieting has received relatively little attention from mainstream researchers of eating disorders. The purpose of the present article is to highlight cognitive ('top-down') factors that are considered responsible for anticipatory anxiety of stoutness and frank fat-phobia (laparophobia). A cognitive model proposed departs from the formulation suggesting that phobia of over-eating is superimposed on avoidant tendencies ('environmental autonomy syndrome'), whereas excessive exercising becomes a natural coping strategy with laparophobia, an instrument of reward. AN ideation involves complex neuronal circuitries and multiple neurochemical components that may conceivably represent a mirror image of those underlying obesity. The emphasis on phobia and aberrant membrane excitability akin to channelopathies behoves the clinicians to be aware of potential uses of drugs acting at the gamma-aminobutyric acid and the N-methyl-D-aspartate/AMPA [2 amino-3-(3-hydroxy-5-methylisoxazol-4-yl) propionic acid] receptors sites as the adjuncts to conventional agents in managing AN. PMID- 16045638 TI - The role of meal replacements in obesity treatment. AB - Effective weight loss strategies are needed to help overweight and obese people lose weight and maintain weight loss in the long term. Meal replacements for weight loss are available in the community; however, their efficacy in the longer term is not clear. This review summarizes recent evidence on the use of meal replacements as a weight loss strategy. PMID- 16045639 TI - Safety of low-carbohydrate diets. AB - Low-carbohydrate diets have re-emerged into the public spotlight and are enjoying a high degree of popularity as people search for a solution to the population's ever-expanding waistline. The current evidence though indicates that low carbohydrate diets present no significant advantage over more traditional energy restricted diets on long-term weight loss and maintenance. Furthermore, a higher rate of adverse side-effects can be attributed to low-carbohydrate dieting approaches. Short-term efficacy of low-carbohydrate diets has been demonstrated for some lipid parameters of cardiovascular risk and measures of glucose control and insulin sensitivity, but no studies have ascertained if these effects represent a change in primary outcome measures. Low-carbohydrate diets are likely effective and not harmful in the short term and may have therapeutic benefits for weight-related chronic diseases although weight loss on such a program should be undertaken under medical supervision. While new commercial incarnations of the low-carbohydrate diet are now addressing overall dietary adequacy by encouraging plenty of high-fibre vegetables, fruit, low-glycaemic-index carbohydrates and healthier fat sources, this is not the message that reaches the entire public nor is it the type of diet adopted by many people outside of the world of a well designed clinical trial. Health effects of long-term ad hoc restriction of inherently beneficial food groups without a concomitant reduction in body weight remains unanswered. PMID- 16045640 TI - Conjugated linoleic acid evokes de-lipidation through the regulation of genes controlling lipid metabolism in adipose and liver tissue. AB - Conjugated linoleic acid (CLA) is a unique lipid that elicits dramatic reductions in adiposity in several animal models when included at < or = 1% of the diet. Despite a flurry of investigations, the precise mechanisms by which conjugated linoleic acid elicits its dramatic effects in adipose tissue and liver are still largely unknown. In vivo and in vitro analyses of physiological modifications imparted by conjugated linoleic acid on protein and gene expression suggest that conjugated linoleic acid exerts its de-lipidating effects by modulating energy expenditure, apoptosis, fatty acid oxidation, lipolysis, stromal vascular cell differentiation and lipogenesis. The purpose of this review shall be to examine the recent advances and insights into conjugated linoleic acid's effects on obesity and lipid metabolism, specifically focused on changes in gene expression and physiology of liver and adipose tissue. PMID- 16045641 TI - Fuel sensing and the central nervous system (CNS): implications for the regulation of energy balance and the treatment for obesity. AB - This review describes the product of the 3-day International Association for the Study of Obesity (IASO) Stock Conference held in March 2004 and sponsored by Abbott Laboratories. The conference was focused on how the mechanisms by which individual cells sense their own fuel status might influence the energy balance of the entire organism. Whether you are a single-celled organism or a sophisticated mammal with a large cerebral cortex, it is critical that cellular activity be matched to the available fuel necessary for that activity. Rapid progress has been made in the last decade in our understanding of the critical metabolic events that cells monitor to accomplish this critical task. More recent developments have begun to apply this understanding to how critical populations of neurones may monitor similar events to control both food intake and energy expenditure. The picture that emerges is that numerous peripheral fuel sensors communicate to the central nervous system (CNS) via neural and humoral routes. Moreover, it has been known for decades that specific populations of neurones sense changes in ambient glucose levels and adjust their firing rate in response and changes in neuronal glucose metabolism can influence energy balance. The CNS, however, does not just sense glucose but rather appears to be sensitive to a wide range of metabolic perturbations associated with fuel availability. This information is used to adjust both caloric intake and the disposition of fuels in the periphery. Increased understanding of these CNS fuel-sensing mechanisms may lead to novel therapeutic targets for obesity. PMID- 16045643 TI - How to learn from adverse events? PMID- 16045644 TI - Frontiers in Scandinavian anaesthesiology. PMID- 16045645 TI - Some aspects of Nordic anesthesiology: past, present and future. PMID- 16045646 TI - Team communication in the operating room. AB - Good communication is vital for safe patient care and good team functioning, not only in the Operating Room but also in all areas of healthcare, as well as in other safety critical industries. Examples from aviation demonstrate both the failures and the successes that can arise from poor and excellent communication. There are six components of effective teamwork: situational awareness, problem identification, decision making, workload distribution, time management and conflict resolution. Practising these, and self-evaluation of team communication, should help to improve team function and contribute to making patient care safer. PMID- 16045647 TI - Genetic variability and clinical efficacy of morphine. AB - The individual variability of opioid pharmacology suggests that the patients' genetic disposition influences the response to opioids. Given the complexity of morphine pharmacology, variability may be caused by several genes. We review data which shows that variability in genes coding the enzyme metabolizing morphine (UGT2B7 gene), mu-opioid receptors (OPRM gene) and blood-brain barrier (BBB) transport of morphine by multidrug resistance transporters (MDR1 gene) influences the clinical efficacy of morphine. Furthermore, variability in an enzyme degrading catecholamines (COMT gene) alters the efficacy of morphine demonstrating that genetic variability in non-opioid systems may indirectly influence the clinical efficacy from morphine. Thus, results obtained so far strongly argue that opioid efficacy is partly related to inborn properties caused by genetic variability. PMID- 16045648 TI - The future of CPR. PMID- 16045649 TI - Future of regional anaesthesia. AB - The development and refinement of regional anaesthetic techniques for various types of surgery, mainly obstetric, ophthalmic and orthopaedic surgery, and of continuous regional analgesia continues. Suitable analgesic drug mixtures, and concentrations, will be further tested in order to find the ideal analgesic regimen for each type of surgery and for the individual patient. No new local anaesthetics or equipment for clinical use are expected in the near future. Improvement therefore depends much on how the anaesthesiologists use the present drugs, needles, nerve detection devices, catheters and pumps. During training in regional anaesthesia for the speciality of anaesthesiology and intensive care medicine, it may suffice to concentrate only on certain common techniques such as epidural block, spinal block, axillary brachial plexus block, intravenous regional anaesthesia and femoral nerve block. Rare regional anaesthetic blocks and invasive techniques should be mastered and taught by specially trained regional anaesthesiology experts. In chronic pain, regional anaesthetic blocks with local anesthetics are not expected to play any major therapeutic role. However, nerve blocks can be useful for diagnostic purposes and in order to facilitate rehabilitation in chronic pain syndromes. PMID- 16045650 TI - Follow-up after intensive care: another role for the intensivist? PMID- 16045651 TI - The future role of the anaesthesiologist in pain management. AB - BACKGROUND: Relieving pain during and after surgery and trauma has always been a basic duty of anaesthesiologists. With their skills and expertise in regional analgesia and pharmacological analgesia, anaesthesiologists have improved management of severe cancer pain. Will there be a place for anaesthesiologists in multidisciplinary pain clinics managing chronic non-cancer pain patients in the future? METHODS: This is a personal review of the development of pain management as a growing part of the responsibilities of anaesthesiologists during the past three decades and the importance of continued involvement of anaesthesiologists in this interesting and challenging aspect of clinical medicine. RESULTS AND CONCLUSIONS: Optimal management of pain during and after surgery is a prerequisite for successful short- and long-term rehabilitation after surgery. After surgery, reducing dynamic pain with prolonged optimal epidural analgesia and regional blocks facilitates mobilization and reduces chronic pain. The expertise of well-trained anaesthesiologists in skillfully using regional analgesia and pharmacological pain relief continues to be in demand in palliative care. Some interventional techniques are useful in relieving chronic non-cancer pain in selected patients. Well-trained anaesthesiologist-pain clinicians can perform interventional treatments safely. No doubt, anaesthesiologists will continue to have important roles in pain management in the future. PMID- 16045652 TI - Factorial design provides evidence to guide practice of anaesthesia. AB - Many scientific articles are written merely to get something published, neglecting the clinician who would like the medical literature to guide their practice. Evidence-based medicine is expected to help in clinical decision making. Systematic reviews of the literature followed by a meta-analysis of randomized, controlled trials (RCT) have claimed to represent the highest strength of evidence. However, the results published in meta-analyses have not always been confirmed in subsequent large RCTs. An analysis of 12 large RCTs and 19 meta-analyses addressing the same questions found that the outcomes of these large RCTs were not predicted accurately 35% of the time by previously published meta-analyses. Therefore, meta-analyses of several small RCTs do not obviate the need for large, multicentre RCTs, which can still be considered as a gold standard for the development of clinical guidelines or practice plans. Moreover, large RCTs using a factorial design can be highly efficient because they can answer several clinical questions at the same time and offer the only systematic approach to investigate an interaction of combinations in multimodal approaches. PMID- 16045653 TI - Identification and characterization of errors and incidents in a medical intensive care unit. AB - BACKGROUND: To assess the frequency, type, consequences, and associations of errors and incidents in a medical intensive care unit (ICU). METHODS: Two-hundred and sixteen consecutive patients with predominantly cardiovascular and pulmonary disorders admitted between December 2002 and February 2003 were enrolled. Demographic data, SAPS II, and TISS-28 were obtained for all patients. Prior to patient enrolment all staff members (physicians, nurses, physiotherapists) were repeatedly encouraged to make use of the Incident Report Form (IRF) and detailed descriptions on how, why and when to use the IRF were provided. RESULTS: During the observation period of 64 days, 50 errors involving 32 patients (15%) were reported. Patients subjected to errors were more severely ill (SAPS II 42 +/- 25 vs. 32 +/- 18, P < 0.05), had a higher hospital mortality (38% vs. 9%), and a longer ICU stay (11 +/- 18 vs. 3 +/- 5 days, P < 0.05). Gender, age and TISS-28 were equally distributed. Each day of ICU stay increased the risk by 8% (odds ratio 1.078, 95% confidence interval 1.034-1.125, P < 0.001), and by 2.3% per SAPS II point (odds ratio 1.023, 95% confidence interval 1.006-1.040, P < 0.001). The majority of errors and incidents were judged as 'human failures' (73%), and 46 errors and incidents (92%) as 'avoidable'. CONCLUSIONS: The identification and characterization of errors and incidents combined with contextual information is feasible and may provide sufficient background information for areas of quality improvement. Areas with a high frequency of errors and incidents need to undergo process evaluation to avoid future occurrence. PMID- 16045654 TI - Comparison of perioperative spirometric data following spinal or general anaesthesia in normal-weight and overweight gynaecological patients. AB - BACKGROUND: There is limited data comparing the impact of spinal anaesthesia (SA) and general anaesthesia (GA) on perioperative lung function. Here we assessed the differences of these two anaesthetic techniques on perioperative lung volumes in normal-weight (BMI < 25) and overweight (BMI 25-30) patients using spirometry. METHODS: We prospectively studied 84 consenting patients having operations in the vaginal region receiving either GA (n = 41) or SA (n = 43). Both groups (GA and SA) were further divided into two subgroups each (normal-weight vs. overweight). We measured vital capacity (VC), forced vital capacity (FVC), forced expiratory volume in 1 s (FEV1), midexpiratory (MEF25-75) and peak expiratory flow rates (PEFR) at the preoperative assessment (baseline), after premedication, after effective SA, and 20 min, 1 h, 2 h and 3 h after the operation (last measurement after patient mobilization). RESULTS: Premedication was associated with a small but significant decrease in lung volumes in direct correlation with BMI (-5%). Spinal anaesthesia resulted in a significant reduction in lung volumes in overweight as opposed to normal-weight patients. Postoperatively, lung volumes were significantly more reduced following GA than SA as indicated by differences in mean VC (SD) of -12 (6)% vs. -6 (5)% 20 min after the end of the operation in the normal-weight and -18 (5)% vs. -10 (5)% in the overweight patients. There was a significant impact of BMI on postoperative respiratory function, which was significantly more important in the GA group than in the SA group, and recovery of lung volumes was more rapid in the normal-weight patients than in the overweight patients, particularly in the SA group. CONCLUSION: In gynaecological patients undergoing vaginal surgery, the impact of anaesthesia on postoperative lung function as assessed by spirometry was significantly less after SA than GA, particularly in overweight patients. PMID- 16045655 TI - Fluid shift is moderate and short-lived during acute crystalloid hemodilution and normothermic cardiopulmonary bypass in piglets. AB - BACKGROUND: Crystalloids are commonly used as priming solutions during cardiopulmonary bypass (CPB). Consequently, hemodilution is a regular occurrence at the start of a CPB. This study describes the time-course variations of hemodynamic parameters, plasma volume (PV) and fluid exchange following crystalloid hemodilution at start of normothermic CPB. METHODS: Forty-five anesthetized piglets were given 60-min normothermic CPB. Ringer's solution was used as priming solution and maintenance fluid. Fluid input/losses, PV, colloid osmotic pressures (plasma/interstitium), hematocrit, and s-proteins were measured, and fluid extravasation rates (FER) and intravascular protein-masses calculated. RESULTS: Start of CPB resulted in a 25-30% hemodilution. To keep the fluid level of the CPB-reservoir constant after start of bypass, fluid addition [2.08 +/- 0.36 (mean +/- SEM) ml kg(-1) min(-1)] was necessary during the first 5 min. Thereafter the fluid needs to be leveled off [0.17 +/- 0.03 ml kg(-1) min( 1) (10-60 min), P < 0.001]. Fluid extravasation rate increased immediately following hemodilution from a baseline value of 0.08 +/- 0.01 to 1.75 +/- 0.34 ml kg(-1) min(-1) with a delayed decrease compared to fluid additions, to reach a 'steady-state' level of 0.22 +/- 0.03 ml kg(-1) min(-1) after 30 min (P < 0.001). Differences in time-course variations between fluid added and fluid extravasated were accompanied by changes in PV and mean arterial pressure. The colloid osmotic gradient decreased about 50% throughout the study and could partly explain the increased FER. CONCLUSION: Acute crystalloid hemodilution contributes to fluid overload during normothermic CPB. The resulting increase in fluid extravasation is, however, moderate, short-lived and levels off to baseline values within 30 min. PMID- 16045656 TI - Changes in jugular bulb oxygen saturation during off-pump coronary artery bypass graft surgery. AB - BACKGROUND: The effect of haemodynamic derangement during coronary artery anastomosis in off-pump coronary artery bypass surgery on cerebral blood flow has not been elucidated. Jugular bulb oxygen saturation is a useful indicator of cerebral blood flow provided that the cerebral metabolic rate is constant. This study was designed to evaluate the changes in jugular bulb oxygen saturation during off-pump coronary artery bypass surgery. METHODS: With IRB approval, 48 patients were included. After anaesthesia, an 18-G catheter was introduced into the jugular bulb. Haemodynamic variables and oxygen profiles from gas analysis of jugular bulb blood and arterial blood were obtained: after sternotomy (baseline); at 5 min after the beginning of the anastomosis of the left anterior descending artery, obtuse marginal artery, and right coronary artery; and after sternal closure. RESULTS: Cardiac index and mixed venous oxygen saturation decreased significantly during anastomosis of all three arteries compared to the baseline value. Although the changes in jugular bulb oxygen saturation during anastomosis were statistically significant compared to its baseline value, jugular bulb oxygen saturation remained within normal limit throughout the study. CONCLUSIONS: Jugular bulb oxygen saturation, which represents the global cerebral oxygenation, was well maintained during the anastomosis of all coronary arteries despite significant haemodynamic changes during off-pump coronary artery bypass (OPCAB). PMID- 16045657 TI - Dopamine or norepinephrine infusion during thoracic epidural anesthesia? Differences in hemodynamic effects and plasma catecholamine levels. AB - BACKGROUND: During thoracic epidural anesthesia, an intravenous dopamine infusion augments the systemic pressure response and modifies plasma catecholamine levels. If such an altered response occurs when norepinephrine is infused is not clear. Therefore, dopamine and norepinephrine induced circulatory and catecholamine responses were studied before and during thoracic epidural anesthesia. METHODS: Nine chloralose-anesthetized dogs were equipped with thoracic epidural catheters. Dopamine (5, 10, and 20 microg kg(-1) min(-1)), and norepinephrine (0.1, 0.25, and 0.5 microg kg(-1) min(-1)) were infused before and during epidural anesthesia, while cardiovascular performance and plasma catecholamine changes were studied. RESULTS: Thoracic epidural anesthesia decreased arterial pressure, and cardiac contractility. The systemic pressure response induced by dopamine was augmented during epidural anesthesia. Norepinephrine did not increase arterial pressure and myocardial contractility as markedly as dopamine, and cardiac output was not altered. Thoracic epidural anesthesia attenuated the plasma norepinephrine level. Plasma dopamine levels were augmented by the dopamine infusion during epidural anesthesia, while plasma norepinephrine levels were attenuated. In contrast, norepinephrine augmented the plasma norepinephrine levels during epidural anesthesia. In general, plasma norepinephrine levels were three to six times higher during a norepinephrine infusion compared to a dopamine infusion. CONCLUSION: The cardiovascular response to a graded dopamine infusion is augmented during thoracic epidural anesthesia, while norepinephrine-induced effects are unaltered. The modified plasma catecholamine levels may contribute to the hemodynamic differences between dopamine and norepinephrine infusions during thoracic epidural anesthesia. PMID- 16045658 TI - Renal effects of hypotensive anaesthesia in combination with acute normovolaemic haemodilution with hydroxyethyl starch 130/0.4 or isotonic saline. AB - BACKGROUND: Hypotensive anaesthesia (HA) and acute normovolaemic haemodilution (ANH) are used separately to decrease per-operative blood loss. Reducing blood viscosity by adding ANH to HA may appear profitable in a situation with lowered perfusion pressure and concern about organ ischemia. The aim of this study was to clarify the influence of HA in combination with ANH using crystalloid or colloid as replacement fluid on renal function. METHODS: Hypotensive anaesthesia was induced in 11 patients referred to major spine surgery using sevoflurane in combination with fentanyl/remifentanil. Acute normovolaemic haemodilution was carried out by drawing venous blood into standard blood bags and replacing it by isotonic saline 0.9% (Group S) or HES 130/0.4 (Group V). Renal function was evaluated before, during and up to 8 h after hypotension as the glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) by means of 51Cr EDTA and 125I-Hippuran clearances. RESULTS: Lowering mean arterial blood pressure decreased GFR and ERPF in both groups. During hypotension ERPF was lower in Group S (n = 5) than Group V (n = 6). Renal function was normalized postoperatively. We found a positive but non-significant correlation between the relative GFR change and the duration of hypotension. CONCLUSION: In conclusion, our study demonstrated that renal function, assessed by GFR and ERPF, is transiently reduced during the combination of hypotensive anaesthesia and acute normovolaemic haemodilution. A colloid-based fluid regime (HES 130/0.4) used for haemodilution may preserve renal function to a greater extent than a crystalloid-based regime (0.9% saline). PMID- 16045659 TI - Does ICU length of stay influence quality of life? AB - BACKGROUND: Patients with prolonged stay in the intensive care unit (ICU) use a disproportionate share of resources. However, it is not known if such treatment results in impaired quality of life (QOL) as compared to patients with a short length of stay (LOS) when taking into account the initial severity of illness. METHODS: Prospective, observational case-control study in a university hospital surgical and trauma adult ICU. All patients admitted to the ICU during a 1-year period were included. Patients with a cumulative LOS in the ICU > 7 days, surviving up to 1 year after ICU admission and consenting were identified (group L, n = 75) and matched to individuals with a shorter stay (group S). Matching criteria were diagnostic group and severity of illness. Health-related quality of life (HRQOL) was assessed 1 year after admission using the short-form 36 (SF-36) and was compared between groups and to the general population. Further, overall QOL was estimated using a visual analogue scale (VAS) and willingness to consent to future intensive care, and was compared between groups L and S. RESULTS: Based on ANCOVA, a significant difference between groups L and S was noted for two out of eight scales: role physical (P = 0.033) and vitality (P = 0.041). No differences were found for the physical component summary (P = 0.065), the mental component summary (P = 0.267) or the VAS (P = 0.316). Further, there was no difference in expectation to consent to future intensive care (P = 0.149). As compared to the general population, we found similar scores for the mental component summary and for three of eight scales in group L and five of eight scales in group S. CONCLUSIONS: When taking into account severity of illness, HRQOL 1 year after intensive care is comparable between patients with a short and a long LOS in the ICU. Thus, prolonged stay in the ICU per se must not be taken as an indicator of future poorer HRQOL. However, as compared to the general population, significant differences, mostly in physical aspects of QOL, were found for both groups of patients. PMID- 16045660 TI - Impact of gender on treatment and outcome of ICU patients. AB - BACKGROUND: Gender modifies immunologic responses caused by severe trauma or critical illness. The aim of this study was to investigate the impact of gender on hospital mortality, length of intensive care unit (ICU) stay, and intensity of care of patients treated in ICUs. METHODS: Data on 24,341 ICU patients were collected from a national database. We measured severity of illness with Acute Physiology and Chronic Health Evaluation II (APACHE II) scores and intensity of care with Therapeutic Intervention Scoring System (TISS) scores. We used logistic regression analysis to test the independent effect of gender on hospital mortality. We compared the lengths of ICU stay and the intensity of care of men and women. RESULTS: Male gender was associated with increased hospital mortality among postoperative ICU patients [adjusted odds ratio 1.33 (95% confidence interval 1.12-1.58, P = 0.001)] but not among medical patients [adjusted odds ratio 1.02 (95% confidence interval 0.92-1.13, P = 0.74)]. Male gender was associated with an increased risk of death particularly in the oldest age group (75 years or older) and among the patients with relatively low APACHE II scores (<16). Mean length of ICU stay was 3.2 days for men and 2.6 days for women (P < 0.001). Male patients comprised 61.7% of the study population but consumed 66.0% of days in intensive care. CONCLUSION: Male gender contributes to poor outcome in postoperative ICU patients. Approximately two-thirds of ICU resources are consumed by male patients. PMID- 16045661 TI - Inflammatory response during hyperglycemia and hyperinsulinemia in a porcine endotoxemic model: the contribution of essential organs. AB - BACKGROUND: During euglycemia acute hyperinsulinemia diminishes the cytokine response to endotoxin [Lipopolysaccharide (LPS)] exposure. In this study we elucidated whether acute hyperglycemia and hyperinsulinemia modify the cytokine content in several organs during LPS challenge in a porcine model. METHODS: Pigs (35-40 kg) were randomized to either normoglycemia (group 1, n = 8) or hyperglycemia and hyperinsulinemia (group 2, n = 8), anesthetized and mechanically ventilated. Both groups received a 180-min intravenous infusion of LPS (total 10 microg kg(-1)). Groups 1 and 2 were clamped at plasma glucose concentrations of 5 mM and 15 mM, respectively. Group 1 maintained a baseline insulin level while the hyperglycemic group exhibited increased insulin levels. RESULTS: Circulating cytokines, cytokine mRNA and cytokine protein content were examined in the heart, liver, kidneys, lungs, spleen, adipose and muscle tissue. After LPS exposure, in both groups vast and equal plasma cytokines were elicited by approximately 70-5000-fold. A 10-fold higher level of IL-10, IL-6 and TNF alpha protein was found in kidney tissue compared to the other organs together with a 3-10-fold increase of TNF-alpha in adipose tissue. However, cytokine mRNAs as well as organ function were without statistical difference between the groups. CONCLUSION: Endotoxemia elicited a pronounced cytokine response in both plasma and at organ level. The kidneys and adipose tissue showed the highest cytokine protein content. Acute hyperglycemia apparently counteracts the well-established anti-inflammatory effects of insulin on the inflammatory response in a LPS challenged porcine model. Whether the observation can be extrapolated to more long-term stress-exposure remains to be clarified. PMID- 16045662 TI - Helicopter transport of sick neonates: a 14-year population-based study. AB - BACKGROUND: The Norwegian Helicopter Emergency Medical Service (HEMS) employs anesthesiologists and paramedics who are not formally trained in neonatology to provide stabilization and transport of sick neonates. We describe neonatal transport by HEMS in central Norway and report the outcome. METHODS: Retrospective analysis of HEMS missions during the 14-year period 1988-2001. RESULTS: A total of 252 neonates were transported, indicating a prevalence of 0.90 per 100 newborn. Median response time was 42 min, on-scene stabilization time 38 min and transportation time 30 min. Median gestational age (GA) was 38 weeks and 4.8% of the neonates were <1000 g and/or <28 weeks. Main clinical problems were respiratory disease, asphyxia and malformations; 30 neonates (12%) died within 1 year. No deaths were transport-related. Tube or ventilator problems were noted in seven out of 66 transports of ventilated neonates. Other technical problems were few. Regarding ventilation, oxygenation and circulation, the clinical condition of most neonates improved during transport and the median temperature rose from 36.5 degrees C to 37.0 degrees C. Hypoglycemia (<2 mmol l( 1)) was documented in 19 missions after transport; of these, eight received a glucose infusion. Four neonates might have profited from HEMS-delivered surfactant therapy. DISCUSSION: Helicopter Emergency Medical Service in central Norway provides rapid medical assistance in a wide spectrum of neonatal problems, but more attention should be paid to proper ventilation and prevention of hypothermia and hypoglycemia. PMID- 16045663 TI - Training trauma teams in the Nordic countries: an overview and present status. AB - BACKGROUND: During the last decade there has been an increased interest in the organisation and quality of trauma care in the Nordic countries. Still, most patients are initially cared for at hospitals with low caseloads of severe trauma. More than 200 hospitals offer initial care to trauma patients. Training of trauma teams using simulators or simulated patients has evolved in the same period, as one important factor to overcome lack of practical training. This overview describes the present state of trauma team training in the Nordic countries. METHODS: Members of a Nordic working group on the use of simulation in medicine reviewed present literature on training with simulation and described the present use of team training in their own countries during winter 2004. RESULTS: There is an increasing amount of evidence indicating that training of teams with simulation reduces treatment errors and improves performance. The training activities do not need to be complex, but skilled debriefing seems necessary. Few Nordic hospitals train their trauma teams. The training activities vary considerably between and within countries. CONCLUSION: There is considerable evidence supporting an increased use of experience gained in other high-risk domains where training in communication, leadership and decision-making is the focus for safety and improvement efforts. There is a need for more widespread training of trauma teams. The different training activities actually undertaken should be scientifically evaluated. PMID- 16045664 TI - Is ECG-guidance a helpful method to correctly position a central venous catheter during prehospital emergency care? AB - BACKGROUND: Insertion of a central venous catheter (CVC) in an emergency situation is challenging and may be potentially associated with more complications. Because CVC positioning by ECG-guidance may help to decrease the frequency of a malpositioned catheter, we decided to prospectively evaluate the usefulness of positioning a CVC by ECG-guidance during prehospital emergency care. METHODS: Prospective observational study during which all patients requiring CVC placement during prehospital care were included. We compared two periods of 1 year during which CVCs were inserted without and then with the help of ECG-guidance. RESULTS: Eighty successive patients were included. We observed a significant reduction of incorrectly positioned CVCs with ECG-guidance (13% vs. 38%, P < 0.05) and a decreased number of chest X-rays needed to verify the position of the CVC (40 vs. 54, P < 0.05). CONCLUSION: ECG-guidance is a safe and feasible technique which significantly improved the rate of CVCs correctly positioned during prehospital emergency care. PMID- 16045665 TI - Postpartum neurologic symptoms following single-shot spinal block for labour analgesia. AB - BACKGROUND: As part of a quality assurance program, we investigated the incidence of postpartum neurologic symptoms in multiparous parturients receiving spinal block for labour analgesia, now in routine use in our labour ward. METHODS: Two hundred and twenty-nine consecutive multiparous parturients presenting for vaginal delivery and requesting spinal analgesia were asked to participate in this prospective study. All parturients received our standard intrathecal analgesia (ITA): 2.5 mg bupivacaine (1 ml) + 25 microg fentanyl (0.5 ml) using a 27-gauge Quincke-type needle. The patients filled in a questionnaire on the first day after delivery and again upon discharge. Complaints typical of neurologic sequelae were noted and a neurologic examination was performed, if necessary. All patients with postdural puncture headache (PDPH) and transient neurologic symptoms (TNSs) were interviewed by telephone 2 weeks after discharge to determine the course of the symptoms. RESULTS: Two hundred and twelve parturients were included in the study. Eighteen (8.5%) parturients complained of PDPH, the severity of which was mild in eight (4%), moderate in seven (3%), and severe in three (1%) patients, respectively. Fifteen (7%) mothers were treated with analgesics or bedrest only. Three (1%) patients were given an epidural blood patch. The paramedian approach was associated with the development of PDPH (P = 0.04). Transient neurologic symptoms were experienced by nine (4.2%) mothers, lasting 1-3 days, mostly presenting as bilateral pain in the buttocks or thighs. One parturient suffered from paraesthesia of the left foot lasting for 3 days. Forty (19%) mothers complained of non-postural headache and 28 (13%) of new-onset back pain. Three mothers (1%) would not want to receive a further spinal block. CONCLUSION: Transient neurologic symptoms (TNSs) after spinal block occurred infrequently. The incidence of PDPH was higher than in the obstetric population in general and calls for re-evaluation of our spinal block methods. Despite the occurrence of neurologic sequelae, patient acceptability was high. PMID- 16045666 TI - Single-shot spinal block for labour analgesia in multiparous parturients*. AB - BACKGROUND: Intrathecal analgesia (ITA) is effective in late, rapidly progressing labour. In 1998, our hospital implemented the use of single-shot spinal block for pain relief in multiparous parturients. As part of a quality assurance programme, we assessed the analgesic efficacy, obstetric and neonatal outcomes and maternal satisfaction after this form of analgesia now in routine use in our labour ward. METHODS: Two hundred and twenty-nine consecutive multiparous parturients presenting for vaginal delivery and requesting analgesia were asked to participate in this prospective study. All parturients received our standard ITA: 2.5 mg bupivacaine (1 ml) + 25 microg fentanyl (0.5 ml) inserted via the L2-3 or L3-4 interspace. Routine monitoring included maternal vital signs, uterine contraction and fetal heart rate tracing. Pain scores (visual analogue scale (VAS) 0-10), sensory levels, motor block, side-effects and maternal satisfaction were recorded. Satisfactory analgesia was defined as a decrease in pain scores to three or less within 20 min after injection. The number of parturients requesting additional analgesia and the duration of spinal block (time from injection until request for additional medication after satisfactory analgesia had worn off) were recorded. RESULTS: Two hundred and nine parturients were included in the study. Satisfactory analgesia was achieved in 153 (73%) parturients. Fifty-five (26%) women requested additional analgesia: 38 (18%) because of unsatisfactory analgesia and 17 (8%) because analgesia ended before delivery. The duration of spinal block was 101 +/- 34 min. Pruritus occurred in 64%, fetal bradycardia in 7% and hypotension in 2% of deliveries. Pain relief was rated excellent by 65%, moderate by 20% and inadequate by 14% of the parturients. One hundred and sixty nine (81%) women stated that they would like to have spinal analgesia again for pain relief during delivery. CONCLUSION: The majority of multiparous parturients found ITA adequate for pain relief during delivery. However, modifications are required in terms of improved timing, reliability and duration of analgesia. PMID- 16045667 TI - Infraclavicular block causes less discomfort than axillary block in ambulatory patients. AB - BACKGROUND: This randomized study was designed to compare discomfort caused by axillary or infraclavicular blocks in ambulatory patients. We identified which of the three block components, needle passes, local anesthetic (LA) injections, and electrical stimulations, is most painful and quantified pain intensity on a visual analog scale (VAS 0-100). We also assessed onset and quality of analgesia, adverse events and patients' acceptance. METHODS: Eighty patients were studied. In axillary group-A, four LA injections were made after stimulating median, musculocutaneous, ulnar and radial nerves. In infraclavicular group-I, the whole LA volume was injected after stimulating median or ulnar or radial nerves. Patients were ready for surgery when they had analgesia/anesthesia distal to the elbow. RESULTS: Median intensity of block discomfort was 22 in A group and 10 in I group (P < 0.01). There was no difference in distribution of the most painful block components between the groups. Block performance times were 4 min in I group and 7 min in A group (P < 0.01). Block onset times were 18 min in A group and 20 min in I group (NS). There was one block failure in I group. Three patients in A group and five in I group required supplementary blocks (NS). Transient adverse events occurred in 14 A-group and two I-group patients (P<0.01). Thirty-seven I-group and 33 A-group patients were satisfied with the block (NS). CONCLUSIONS: Infraclavicular block by single injection caused less discomfort and fewer adverse events than axillary block by multiple injections. Block effectiveness, onset time and patients' acceptance were similar. PMID- 16045668 TI - Comparison of two different techniques for brachial plexus block: infraclavicular versus axillary technique. AB - BACKGROUND: Brachial plexus block via the axillary approach is problematic in patients with limited arm mobility. In such cases, the infraclavicular approach may be a valuable alternative. The purpose of our study was to compare axillary and infraclavicular techniques for brachial plexus block in patients undergoing arm or forearm surgery. METHODS: After institutional approval and informed consent were obtained, 30 patients (ASA physical status I or II) scheduled for forearm and hand surgery under brachial plexus anesthesia were included in the study. Patients were randomly allocated into two groups. Brachial plexus block was performed via the axillary approach in the Group A patients and via the infraclavicular approach in the Group I patients using a peripheral nerve stimulator. All blocks were performed with a total dose of 40 ml 0.375% bupivacaine. RESULTS: In each nerve territory (radial, ulnar, median, and musculocutaneous), the mean values of the degree and the duration of the sensory block and motor block were not significantly different between the two groups (P > 0.05). Inadvertent vessel puncture was significantly more frequent in the axillary approach (P < 0.05). CONCLUSION: Brachial plexus block performed via the infraclavicular approach is as safe and effective as the axillary approach. Infraclavicular approach may be preferred to the axillary approach when the upper arm mobility is impaired or not desired. PMID- 16045669 TI - Itching for progress. AB - The study of itch remains a neglected field, with a number of barriers limiting satisfactory therapy in the majority of instances. We review recent advances, focusing on the identification of the neural pathways, distinct from pain, that signal itch; methods to measure itch as scratch in humans; and the identification of a role for H4 receptors in mediating itch (in the mouse). PMID- 16045670 TI - Incontinentia pigmenti case series: clinical spectrum of incontinentia pigmenti in 53 female patients and their relatives. AB - A retrospective case series of 53 female patients with incontinentia pigmenti (IP) including 28 secondary cases (female relatives of probands) was reviewed and compared with other series in an attempt to estimate more accurately the true disease burden of patients with IP. We found that, while the frequency of the first three cutaneous stages of IP was comparable with previous studies, none of the secondary cases manifested any serious neurological complications but all displayed stage IV pale anhidrotic reticulate lines on their posterior calves. This important clinical feature of IP in secondary cases has previously been under-represented in studies that often involved only paediatric probands. Hence, mildly affected cases of IP are often undiagnosed and under-represented in case series to date, possibly leading to inappropriately high estimates of neurological and eye involvement. With the availability of genetic testing, it is now feasible to confirm the variability of the phenotype and the risk of complications in IP. PMID- 16045671 TI - Vulval squamous cell carcinoma arising in chronic hidradenitis suppurativa. AB - We report a case of vulval squamous cell carcinoma (SCC) arising in chronic hidradenitis suppurativa (HS). The patient had a complex medical history including a 25-year-history of Crohn's disease. In addition she had recently received immunosuppressive therapy for nephrotic syndrome secondary to membranous glomerulonephritis. A painful nodule was noted on the vulva that was clinically very suspicious of SCC. An excision biopsy confirmed the diagnosis. There are few publications in the English literature citing association between HS and the development of SCC. The first report in the English literature of vulval SCC arising in chronic HS was published in 1999. We wish to draw attention to the possibility that patients with HS may develop SCC in lesional skin. A painful lump or ulcer could easily be mistaken for an inflammatory lesion and a low threshold for biopsy is warranted. We suggest constant vigilance with regard to malignant change in ano-genital HS as the diagnosis can be difficult. PMID- 16045672 TI - Cutaneous calcification following liver transplantation. AB - Ectopic calcification following liver transplantation has been reported to occur in various internal organs but there have been few reports of skin involvement. The pathogenesis is uncertain with previous reports suggesting that the calcifications could be either dystrophic or metastatic. The large amount of intravenous calcium needed to correct hypocalcaemia secondary to blood product transfusion is thought to play a central role. We report a case of calcinosis cutis developing after liver transplantation in a 22-year-old woman at sites where no intravenous calcium had been administered. In previously published cases serum calcium and phosphate levels were reported as normal. In our case serum calcium levels were also within or below normal limits with the exception of a transient rise in the immediate post-operative period. Our case supports earlier hypotheses that short-lived and often undetected elevations in the calcium phosphate product are implicated in this condition. PMID- 16045673 TI - Antibiotic prophylaxis in patients with valvular heart defects undergoing dermatological surgery remains a confusing issue despite apparently clear guidelines. AB - The guidelines pertaining to prescription of prophylactic antibiotics to prevent endocarditis during dermatological surgery appear clear and well-documented. The British Society for Dermatological Surgery, in agreement with the British Society for Antimicrobial Chemotherapy, state that antibiotic prophylaxis for endocarditis is not required for routine dermatological surgery procedures even in the presence of a pre-existing heart lesion. Pre-existing cardiac lesions include prosthetic valves, history of bacterial endocarditis, congenital cardiac malformation, rheumatic or other acquired valvular dysfunction, hypertrophic cardiomyopathy or mitral valve prolapse with regurgitation. It is important to distinguish between antibiotic prophylaxis for wound infection and that for bacterial endocarditis. Routine procedures, such as punches, shaves, curettage and simple excisions, performed on clean intact skin have an extremely low risk of wound infection (1-4%). The risk of wound infection increases to 5-15% with clean-contaminated skin surgery that includes procedures involving eroded or ulcerated skin, respiratory or buccal mucosa, flexural areas and protracted procedures such as Mohs' micrographic surgery. In such cases, antibiotic prophylaxis may be considered in patients with a cardiac lesion because a wound infection may result in bacteraemia and subsequent endocarditis. This should therefore not be considered 'routine' dermatological surgery. In contaminated, dirty and/or infected classes of wounds the risk of wound infection is higher (> 25%). Elective skin surgery should be postponed if possible until the wound infection is treated with therapeutic antibiotics. PMID- 16045674 TI - Acinic cell carcinoma on the lower lip resembling a mucocele. AB - A 64-year-old woman presented with a 2-week history of an asymptomatic mass involving the lower lip. Histopathological examination showed a well circumscribed tumour composed of many lobules separated by thin, fibrous connective tissue. Individual lobules were composed of round or polyhedral tumour cells, which had a characteristic finely granular and vacuolated cytoplasm and eccentric hyperchromatic nuclei. Positive staining was observed with Periodic acid-Shiff, and immunohistochemistry for cytokeratin, alpha-1 antitrypsin, and S 100 protein resulting in a final diagnosis of acinic cell carcinoma. Acinic cell carcinoma represents a well-established, although uncommon, entity in the classification of neoplasms of salivary gland origin. The parotid salivary gland is the most frequent site of acinic cell carcinoma, whereas the lip is a particularly unusual site. The unusual presentation of this tumour may lead to confusion with a mucocele. Given these findings, we suggest that acinic cell carcinoma should be considered in the differential diagnosis of any mucocele-like mass on the lower lip. PMID- 16045675 TI - Dyschromatosis universalis hereditaria: two cases in a Chinese family. AB - Two cases of dyschromatosis universalis hereditaria (DUH) from a Chinese family are presented. Case 1 was a 62-year-old woman who had a generalized and progressive hyper- and hypopigmentation of the skin from the age of 8 years. Her brother had also developed a similar skin pigmentary defect from about the same age. Histopathological and ultrastructural examination of lesional skin showed increased melanin content in epidermal keratinocytes but no changes in the appearance or number of melanocytes. Although hereditary defects may influence melanogenesis resulting in a pigmentary anomaly, the pathogenesis of DUH remains unclear. PMID- 16045676 TI - Two cases of atypical membranous aplasia cutis with hair collar sign: one with dermal melanocytosis, and the other with naevus flammeus. AB - We report two atypical cases of membranous aplasia cutis surrounded by a rim of hairs, one case associated with dense dermal melanocytosis and the other with naevus flammeus, with characteristic clinical features. A rim of hypertrichosis, 'hair collar' sign, is proposed to have a close association with neuroectodermal defects. A failure of the normal closure of the cranial neural tube might have affected foetal skin development, including melanoblast migration and capillary network formation. The changes in the present cases, as well as the hair collar sign may suggest a complex hamartomatous nature of membranous aplasia cutis. PMID- 16045677 TI - Persistent severe amiodarone-induced photosensitivity. AB - Amiodarone, a benzofuran derivative, has been used therapeutically as an antiarrhythmic and coronary vasodilator in Europe since 1964. One of its commoner side effects is cutaneous photosensitivity; more rarely, after ingestion of the drug for around 12 months, a slate-grey or violaceous discoloration of sun exposed sites may gradually develop. Both of these side effects usually resolve within 2 years of discontinuation of the drug. We now present a woman who developed both photosensitivity and a slate-grey discoloration whilst taking amiodarone; on discontinuation of the drug, the dyspigmentation gradually resolved, but the photosensitivity has persisted and the patient remains symptomatic more than 17 years later. PMID- 16045678 TI - A case of bullous pemphigoid associated with autoantibodies targeting antigenic sites other than the NC16a domain of BP180. AB - Bullous pemphigoid (BP) is associated with autoantibodies to the 180-kDa BP antigen (BP180), and the antigenic site exists on noncollagenous 16a (NC16a) domain of BP180. We now report a male BP patient whose IgG autoantibodies did not react against the NC16a domain of BP180 by either immunoblotting or ELISA, whereas they did react with BP180 extracted from normal human keratinocytes. Anti BP180 cicatricial pemphigoid was ruled out due to the lack of conjunctival mucosal involvement and the absence of scarring in the oral cavity. Our findings indicate that there is an antigenic reactive region other than NC16a on the extracellular domain of BP180. There have been few reports describing detailed clinical features of BP caused by autoantibodies targeting antigenic sites other than the NC16a domain. We conclude that it is difficult to differentiate their clinical features from those associated with autoantibodies targeting the NC16a domain of BP180. PMID- 16045679 TI - Invasive dermatophytosis with lymph node involvement in an immunocompetent patient. AB - A 23-year-old man presented with annular and arcuate, hyperpigmented, itchy, scaly plaques over the trunk and lower extremities for 5 years progressing to verrucous papules and nodules for the last 1.5 years. He also had nontender, inguinal and axillary lymphadenopathy. Skin and lymph node biopsies showed granulomatous inflammation and special stains demonstrated long septate hyphae. Tissue cultures grew Trichophyton verrucosum. The patient was treated with itraconazole 100 mg twice daily for 8 months, resulting in complete clearance of the lesions. PMID- 16045680 TI - Folliculosebaceous cystic hamartoma arising within a port-wine stain. AB - A 34-year-old woman presented with 2-year history of a dome-shaped papule on a well-circumscribed, thickened, port-wine stain on the left side of the chin. Squeezing on the port-wine-stain plaque revealed many comedos within dilated follicular orifices. The papule was excised and submitted for histological examination. Histopathological study showed a lobular neoplasm, comprising dilated, cystic pilosebaceous structures surrounded by fibrous stroma, bearing the characteristics of folliculosebaceous cystic hamartoma. The reported case shows that, in addition to the vascular nature, both ectodermal and mesenchymal abnormalities may be involved in port-wine stains. PMID- 16045681 TI - Seasonal occurrence of impetigo: a retrospective 8-year review (1996-2003). AB - Impetigo, a common skin infection, has shown seasonal variation in African, Australian and Indian studies. We investigated seasonal variation of impetigo in a UK paediatric population. A total of 1552 children with impetigo were seen in the Accident and Emergency (A&E) department between 1996 and 2003. The number of impetigo cases was always higher in late summer than in winter, and furthermore, increased year on year. These changes could not be accounted for by variation in total patient numbers seen in A&E, and suggest a correlation between impetigo frequency and climatic temperature. Possible reasons for these findings include exposed skin due to loose clothing in the summer leading to more skin-to-skin contact and minor trauma. PMID- 16045682 TI - A case of multicentric reticulohistiocytosis with multiple lytic skull lesions. AB - Multicentric reticulohistiocytosis (MR) is a rare multisystemic disorder of unknown aetiology characterized by cutaneous and joint manifestations. It is associated with malignancy in up to 31% of cases. Common radiological findings are peri-articular erosions and osteolytic punched-out areas. We present a case of MR with cutaneous nodules, joint pains, and multiple lytic skull lesions--a combination that has not been described before. Osteolytic activity of proinflammatory cytokines (tumour necrosis factor-alpha and interleukin-1) may explain the peri-articular erosions often seen in MR, and the multiple lytic skull lesions seen in our patient. PMID- 16045683 TI - Successful adjuvant treatment of severe bullous pemphigoid by tryptophan immunoadsorption. AB - Bullous pemphigoid (BP) is an autoimmune blistering skin disease associated with circulating autoantibodies to the hemidesmosomal antigens BP180 and BP230. We report two cases of therapy-refractory BP adjuvantly treated by tryptophan immunoadsorption. In both patients, this treatment was associated with rapid clinical improvement and reduction in the required corticosteroid dosage. In addition, levels of circulating anti-BP180 autoantibodies decreased markedly. Antibodies that were eluted from the tryptophan matrix bound to BP180 and induced dermal-epidermal separation in cryosections of human skin. Our observations suggest that immunoadsorption may be a helpful adjuvant treatment in severe BP. PMID- 16045684 TI - A case of acquired smooth muscle hamartoma of the scrotum. AB - We report a case of acquired smooth muscle hamartoma (ASMH) of tunica dartos, believed to be only the fifth so far reported. A 24-year-old man had a 6-month history of an asymptomatic tight and thickened scrotum. The skin was difficult to pinch. A biopsy specimen from the skin showed increased and proliferated smooth muscle bundles composed of well-differentiated and uniform spindle cells that showed typical features of acquired smooth muscle hamartoma. Interestingly, dilatations of the lymph vessels were noted in the upper dermis above the proliferated smooth muscles. It has been reported recently that long-standing severe lymphoedema may cause histological features mimicking ASMH. As the present case was not preceded by oedema of the scrotum, we consider this case to be true ASMH. PMID- 16045685 TI - Lucio phenomenon and Lucio leprosy. AB - Lucio phenomenon is a peculiar reactional state associated with Lucio leprosy; both exhibit a restricted global distribution. The exact underlying pathomechanism of Lucio phenomenon, which may be fatal at times, still needs further elaboration. A case of relapse of partially treated nodular lepromatous leprosy presenting with Lucio phenomenon is reported, along with a brief review of the literature. PMID- 16045686 TI - A case of Schopf-Schulz-Passarge syndrome. AB - Schopf-Schulz-Passarge syndrome (SSPS) is a rare ectodermal dysplasia characterized by hypodontia, hypotrichosis, nail dystrophy, palmoplantar keratoderma, and periocular and eyelid margin apocrine hidrocystomas. Several other skin tumours have been described in association with this syndrome, in particular, multiple palmoplantar eccrine syringofibroadenoma (ESFA). We report a case of SSPS with diffuse palmoplantar hyperkeratosis, which was shown by histology and immunocytochemistry to be due to ESFA. PMID- 16045687 TI - Inflammatory epidermolysis bullosa acquisita with coexistent IgA antibodies to plectin. AB - We present a case of inflammatory epidermolysis bullosa acquisita (EBA) with IgA antibodies to plectin. Analysis of lesional skin biopsies by electron microscopy revealed the split level to be in the sublamina densa zone, corresponding to the diagnosis of EBA. Direct immunofluorescence of perilesional skin demonstrated u serrated depositions of IgG and IgA that under immunoelectron microscopy were shown to be located in the sublamina densa. In contrast, indirect immunofluorescence on salt-split skin revealed circulating IgA antibodies that stained the roof rather than the floor of the blister. Immunoblotting showed these serum antibodies to be directed to the cytoplasmic hemidesmosomal antigen plectin. The antiplectin specificity of these antibodies was confirmed by 'knockout' immunofluorescence analysis; the serum IgA did not bind to skin sections of a patient with plectin-deficient epidermolysis bullosa. To our knowledge, this case demonstrates for the first time the existence of IgA antibodies against plectin. PMID- 16045688 TI - Direct characterization of human T cells in pemphigus vulgaris reveals elevated autoantigen-specific Th2 activity in association with active disease. AB - Pemphigus vulgaris (PV) is a blistering skin disorder mediated by autoantibodies targeting the epidermal adhesion molecule desmoglein 3 (Dsg3). As Th2-associated cytokines are necessary for directing antibody production, it is hypothesized that Dsg3-specific Th2 activity is associated with active disease. We used cell surface-matrix technology in combination with flow cytometry to characterize the Dsg3-reactive T-cell population using peripheral blood mononucleocytes sampled from PV patients stratified by active (n = 9) or remittent disease (n = 6), and healthy human leucocyte antigen-matched controls (n = 5). We evaluated interferon gamma-producing CD4+ cells (Th1) and interleukin (IL)-10- or IL-4-producing CD4+ cells (Th2). The mean frequency of Th2 CD4+ T cells was significantly elevated for five of nine PV patients with active disease. No significant Th2 responses were detected for patients with remittent disease or controls. There was a significant association of Th2 activity with active disease compared with remittent and control groups (P = 0.026 and P =0.012, respectively), and Th2 activity was significantly correlated with anti-Dsg3 IgG titre (P = 0.044). One patient with remittent disease converted from a Th2-negative to a Th2-positive response with the initiation of disease activity. An antigen-specific CD4- lymphocyte response was detected in five PV patients (36%), and was shown to correlate closely with the CD8+ population. These results are consistent with the hypothesis that Th2 response directs autoantibody production and is therefore associated with disease activity in PV. PMID- 16045689 TI - Expression and serum levels of MMP-2 and MMP-9 during human melanoma progression. AB - Matrix metalloproteinases (MMP)-2 and -9 have been implicated in malignant tumour progression, partly because they degrade collagen type IV, a major component of basement membranes. Biopsy specimens from 56 patients with primary melanoma and 7 with cutaneous or nodal metastases were studied by immunohistochemistry. Of 39 patients with estimated good prognosis, 70.5% of melanomas were negative for MMP 2, compared with only 47% of 17 melanomas in patients who developed metastasis during the 3-year follow-up. All skin and nodal metastases were negative for MMP 2 and positive for MMP-9. Of 14 thick melanomas, 9 were mostly positive for MMP-2 expression, suggesting a possible association with the invasiveness of the melanoma. MMP-2 and MMP-9 plasma levels were analysed in another 29 patients with melanoma (10 stage I and II, 9 stage III, and 10 stage IV) and in 10 healthy controls. No difference in MMP-9 plasma levels was found among the groups. Higher MMP-2 concentrations were observed in patients with metastatic disease (stage IV) than in those with primary melanoma (stage I) or in controls. Serial levels in two patients who passed from stage I to stage III or IV showed no significant difference in MMP-2 or -9 values. We conclude that MMP-2 expression might be associated with progression of the melanoma. Circulating MMP-2 and -9 levels have shown low sensitivity and specificity, so they do not seem to be good tumour markers in patients with melanoma. PMID- 16045690 TI - CTLA-4 gene 49A/G polymorphism in Turkish patients with Behcet's disease. AB - Genetic factors predisposing individuals to Behcet's disease (BD) are considered to play important roles in the development of the disease. Patients with BD exhibit elevated levels of pro-inflammatory cytokines, and affected organs show a significant neutrophil and lymphocyte infiltration. Current evidence suggests that the activated lymphocytes contribute to neutrophil and endothelial cell activation in these patients. The cytotoxic T lymphocyte-associated antigen (CTLA)-4 molecule plays an important role in immune regulation by downregulating T-cell activation, and the CTLA-4 49A/G polymorphism in the exon 1 has been shown to be associated with a number of autoimmune diseases. In an attempt to demonstrate whether there is an association of the CTLA-4 49A/G polymorphism with BD in the Turkish population, we genotyped 59 Turkish patients and 99 healthy individuals for single-nucleotide polymorphisms. For this purpose, genomic DNA was obtained from the peripheral blood of individuals and the region of interest was amplified using PCR. Genotyping was performed using the BbvI restriction endonuclease. It was shown that the distribution of the CTLA-4 exon 1 49A/G allele and genotype frequencies did not differ between patients with BD and healthy controls. However, allele and genotype frequencies of CTLA-4 49 A and A/A were significantly higher in patients with ocular involvement compared with patients without these symptoms (90.6% vs. 65.1%, odds ratio (OR) = 9.67, P = 0.011; and 81.25% vs. 39.5%, OR = 9.56, P = 0.015, respectively). A statistically significant difference in the A allele frequency was observed in patients with erythema nodosum-like lesions (86.1% vs. 65.8%, OR = 6.24, P = 0.04). There was also an increase in A/A genotype frequency, but the difference was not statistically significant (72.2% vs. 41.5%, OR = 6.5, P = 0.068). Our data suggest that BD patients with ocular involvement and erythema nodosum-like lesions have a higher frequency of both the A allele and the A/A genotype at position 49 of exon 1 of CTLA-4. These results may also indicate that CTLA-4 is a disease-modifying rather than a susceptibility gene for BD. PMID- 16045691 TI - Erosive lichen planus of the vulva: weak circulating basement membrane zone antibodies are present. AB - The objective of this study was to investigate whether circulating basement membrane zone (BMZ) antibodies are present in erosive lichen planus (LP) of the vulva. In total, 56 consecutive women with biopsy-confirmed erosive LP of the vulva were recruited from a vulval clinic in a district general hospital and teaching hospital in Oxfordshire. Indirect immunofluorescence (IgG and IgA) was performed on 56 sera, and 15 were tested to IgG subclasses (1-4). Immunoblotting was carried out on salt-split and urea-extracted epidermal skin extracts on 11. The main outcome measure was the presence or absence of staining at the BMZ. Of the 56 sera, 34 (61%) had weak (neat or 1 : 5) epidermal-binding BMZ antibodies (25 had IgG, 5 had IgA, 4 had both IgG and IgA). All 15 sera tested to IgG showed epidermal binding to one or more IgG subclasses: IgG1 (7 sera), IgG2 (7), IgG3 (7) and IgG4 (0). Immunoblotting identified IgG antibodies to bullous pemphigoid (BP)180 (10/11) and BP230 (2/11). The majority (61%) of patients with vulval erosive LP had circulating serum IgG BMZ antibodies, chiefly reacting with BP180. There was subclass restriction of the IgG response to IgG1, 2 and 3. The significance of these antibodies is uncertain, but they may be a marker for the disease. PMID- 16045692 TI - Muscarinic cholinergic receptors in the human melanoma cell line SK-Mel 28: modulation of chemotaxis. AB - Primary and metastatic human melanomas express muscarinic receptors. In embryonic tissues, expression of muscarinic receptors is correlated with morphogenesis. The hypothesis has been put forward that muscarinic receptors are involved in morphogenetic movements in the embryo, and in cellular movements in melanoma cells during invasive growth. The purpose of the present study was to characterize the muscarinic receptors in the human melanoma cell line SK-Mel 28 and to test in a Boyden chamber assay whether the chemotactic activity towards fibronectin can be influenced by muscarinic stimulation. In Western blots with the monoclonal antibody M35, muscarinic receptors were localized in a strong band at 66 kDa, and in a weak band at 63 kDa. Western blot with M3 subtype specific antibodies reproduced the line at 66 kDa. RT-PCR revealed mRNA for subtypes M3 and M5. These findings suggest that SK-Mel 28 cells express a large number of subtype M3 and a small number of subtype M5 receptors. Microscopic observation of calcium mobilization after muscarinic stimulation indicated that all cells carried functional muscarinic receptors. A standardized chemotaxis assay was established in modified Boyden chambers using fibronectin as chemotactic agent. After addition of carbachol to the upper compartment, an increase of fibronectin induced chemotaxis of approximately 30% was observed, an effect abrogated by atropine. These results demonstrate that muscarinic cholinergic treatment has a modulatory effect on fibronectin-induced chemotaxis in SK-Mel 28 melanoma cells, indicating that the muscarinic system is involved in regulation of cell movement. PMID- 16045693 TI - Micronucleus frequency in the oral mucosa and lymphocytes of patients with Behcet's disease. AB - Behcet's disease (BD) is a chronic, multisystemic, inflammatory disorder characterized mainly by recurrent oral and genital aphthous ulcerations and uveitis. Our study aimed to determine the genetic damage in patients with BD. The micronucleus (MN) frequency was counted in peripheral lymphocytes and exfoliated cells of the patients with BD. MN analysis was performed in peripheral lymphocytes of 30 patients with BD and in 20 healthy controls by the cytokinesis block method, and on uncultured cells of the oral cavity in 10 patients and 9 healthy controls. We found significantly higher MN rates in lymphocytes of the patients than the control subjects (P = 0.000). There were no significant differences between the patients with or without treatment (P = 0.860). The MN frequency in exfoliated cells of the patients was higher than in those of healthy controls (P = 0.013), and there was no significant difference between the exfoliated cells of the treated and untreated patients (P = 0.201). Our results indicate that genetic damage may play a secondary but important part in the aetiology of BD and that treatment with colchicine does not induce MN. PMID- 16045694 TI - Anti-inflammatory effects of tretinoin (all-trans-retinoic acid) 0.1% and adapalene 0.1% in rats. AB - In this study, the anti-inflammatory effects of tretinoin (all-trans-retinoic acid) 0.1% cream and adapalene 0.1% gel were compared in rats to determine whether there was a difference between these agents. Thirty-six rats of either sex were divided into six groups (two control groups, and an etodolac, indomethacin, tretinoin and adapalene group) of six animals each. Each group was given different drugs or chemicals. The inhibitory activities of the drugs were determined on carrageenan-induced rat-paw oedema. The inhibition rate (53.48%) in the tretinoin group was found to be higher than adapalene and controls (P < 0.05). Adapalene was found to have an inhibition rate of 10.28%, and when compared with the other groups, was found to have no statistically significant anti-inflammatory activity. We conclude that tretinoin has a higher anti inflammatory activity than adapalene and thus should be preferred for the treatment of inflammatory lesions. PMID- 16045695 TI - A homozygous nonsense mutation in the EVER2 gene leads to epidermodysplasia verruciformis. AB - Epidermodysplasia verruciformis (EV) is a genodermatosis with mainly autosomal recessive inheritance. Pathogenic mutations in two adjacent genes, EVER1 and EVER2, have recently been identified. In this study, we performed mutation detection for the EVER1 and EVER2 genes on samples from a Chinese patient with EV, who had consanguineous parents. A homozygous C-->T transition at nucleotide position 568 within exon 6 of the EVER2 gene was detected. The mutation led to a premature translation termination (R190X) and the predicted protein lacked 537 amino acids. This novel nonsense mutation is, to our knowledge, the first mutation reported in Chinese patients with EV. PMID- 16045696 TI - Novel mutations in the ATP2C1 gene in two patients with Hailey-Hailey disease. AB - Benign familial chronic pemphigus (Hailey-Hailey disease, HHD) is a rare hereditary condition characterized by development of blisters at sites of friction and in the intertriginous areas. Mutations in the ATP2C1 gene, which encodes the human secretory pathway calcium ATPase 1 (hSPCA1), have been identified as possible causative mutations. Studying Hungarian patients with HHD, we found two novel, distinct, heterozygous mutations. In a 65-year-old man with a 41-year history of severe recurrent symptoms, a single nucleotide insertion, 1085insA, was detected. In a patient whose symptoms were induced by environmental contact allergens, we found a nonsense mutation, Q506X, in exon 17. Our study further illustrates the diversity of mutational events in the pathogenesis of HHD. PMID- 16045697 TI - Palisaded neutrophilic and granulomatous dermatitis. PMID- 16045698 TI - Prolonged dermatitis distant to the site of squaric acid dibutyl ester applications and recovery of alopecia areata. PMID- 16045699 TI - Merkel cell carcinoma in a patient with chronic sarcoidosis. PMID- 16045700 TI - Oral cyclophosphamide in a case of cyclosporin and steroid-resistant chronic urticaria showing autoreactivity on autologous serum skin testing. PMID- 16045701 TI - Erythema annulare centrifugum successfully treated with metronidazole. PMID- 16045702 TI - Intractable erythema nodosum associated with severe breast abscesses: reports of two cases. PMID- 16045703 TI - Pubertal giant sebaceous hyperplasia over the nose. PMID- 16045704 TI - Collision of basal cell carcinoma with seborrhoeic keratosis: a dermoscopic aid to histopathology? PMID- 16045705 TI - Acquired port wine stain following oral isotretinoin. PMID- 16045706 TI - Plasma cell balanitis of Zoon treated successfully with topical tacrolimus. PMID- 16045707 TI - Cutaneous presentation of a myxoid cyst occurring at the knee joint. PMID- 16045708 TI - Wrinkly skin syndrome. PMID- 16045709 TI - A role for tubular compression in the management of psoriasis. PMID- 16045710 TI - Remarkable follow-up experiences of a severe persistent case of pemphigoid gestationis. PMID- 16045711 TI - Infantile acquired zinc deficiency resembling acrodermatitis enteropathica. PMID- 16045712 TI - Nodular scabies mimicking urticaria pigmentosa in an infant. PMID- 16045713 TI - Perianal cytomegalovirus ulcer following herpes simplex virus in a patient with idiopathic thrombocytopenic purpura treated with immunosuppressants. PMID- 16045714 TI - Hailey-Hailey disease failing to respond to treatment. PMID- 16045715 TI - Successful treatment of erosive pustular dermatosis of the scalp with topical tacrolimus. PMID- 16045716 TI - The timing of intravenous immunoglobulin therapy in Stevens-Johnson syndrome and toxic epidermal necrolysis. PMID- 16045717 TI - Multiple purpuric lesions on the lower limbs: an unusual presentation of a rare condition at an unusual age. PMID- 16045718 TI - Skin tumours and a family history of cancer. PMID- 16045719 TI - Painful skin induration in a neonate. PMID- 16045720 TI - Herpetic whitlow: a forgotten diagnosis. PMID- 16045721 TI - Tuberculous cold abscess resembling a lymphatic malformation. PMID- 16045726 TI - Immunological profile of peripheral blood lymphocytes and monocytes/macrophages in Kawasaki disease. AB - Kawasaki disease (KD) is an acute illness of early childhood characterized by prolonged fever, diffuse mucosal inflammation, indurative oedema of the hands and feet, a polymorphous skin rash and nonsuppurative lymphadenopathy. The histopathological findings in KD comprise panvasculitis with endothelial necrosis, and the infiltration of mononuclear cells into small and medium-sized blood vessels. The levels of many proinflammatory cytokines, chemokines and adhesion molecules can be elevated in sera from children with KD at the acute stage. Although many immunological studies on KD involving peripheral blood have been reported, the data obtained remain controversial. This review focuses on the immune response of peripheral blood lymphocytes and monocytes/macrophages during acute KD. PMID- 16045727 TI - Insights into the role of fibroblasts in human autoimmune diseases. AB - Traditional wisdom has considered fibroblasts as contributing to the structural integrity of tissues rather than playing a dynamic role in physiological or pathological processes. It is only recently that they have been recognized as comprising diverse populations of cells exhibiting complex patterns of biosynthetic activity. They represent determinants that react to stimuli and help define tissue remodelling through the expression of molecules imposing constraints on their cellular neighbourhood. Moreover, fibroblasts can initiate the earliest molecular events leading to inflammatory responses. Thus they must now be viewed as active participants in tissue reactivity. In this short review, I will provide an overview of contemporary thought about the contribution of fibroblasts to the pathogenesis of autoimmune processes through their expression of, and responses to, mediators of inflammation and tissue remodelling. PMID- 16045728 TI - Histological analysis of CD11c-DTR/GFP mice after in vivo depletion of dendritic cells. AB - To investigate the dependence of individual immunological processes on DC, a transgenic mouse system (CD11c-DTR/GFP mice) has been developed that allows conditional depletion of CD11c+ DC in vivo through administration of diphtheria toxin. We have performed careful histological analysis of CD11c-DTR/GFP mice at different time points after diphtheria toxin injection and confirmed the transient depletion of CD11c+ cells from lymph nodes and spleen. Unexpectedly, the injection of diphtheria toxin completely depleted marginal zone and metallophilic M(Phi) from the spleen and their sinusoidal counterparts from the lymph nodes. This finding limits the use of CD11c-DTR/GFP mice for the analysis of the role of DC to models and read outs that are proven to be independent of marginal zone and sinusoidal M(Phi). PMID- 16045729 TI - Adenovirus-mediated gene delivery of interleukin-10, but not transforming growth factor beta, ameliorates the induction of Graves' hyperthyroidism in BALB/c mice. AB - Interleukin-10 (IL-10) and transforming growth factor-beta (TGF-beta) are well known anti-inflammatory cytokines. We have studied the effect of adenovirus mediated IL-10 and TGF-beta gene delivery on the induction of Graves' hyperthyroidism in our mouse model that involves repeated injections of adenovirus expressing the thyrotropin receptor A subunit (AdTSHR). We first constructed adenoviruses encoding the two cytokines (AdIL10 and AdTGF(beta)) and confirmed expression by in vitro infection of COS cells. Susceptible BALB/c mice were injected twice with AdTSHR alone or together with AdIL10 or AdTGF(beta), and bled two weeks after the second immunization. Significantly elevated serum thyroxine levels were seen in 26% of mice immunized with AdTSHR and AdIL10 versus 61% with AdTSHR alone. Levels of thyroid stimulating antibody, but not nonstimulating antibody, were also decreased, and TSHR-specific splenocyte secretion of interferon-gamma in recall assays was impaired in mice treated with AdIL10. In contrast, AdTGF(beta) had little effect on hyperthyroidism. Overall, our findings demonstrate that gene delivery of IL-10, but not TGF-beta, suppresses the induction of Graves' hyperthyroidism in a mouse model. However, the effect of IL-10 is less powerful than we observed previously with T helper type 2-inducers including adenovirus expressing IL-4, Shistosoma mansoni infection or alpha-galactosylceramide. PMID- 16045730 TI - Macrophage migration inhibitory factor contributes to the development of acute dextran sulphate sodium-induced colitis in Toll-like receptor 4 knockout mice. AB - Toll-like receptor 4 (TLR4), which recognizes lipopolysaccharides, plays an important role in the innate immune response. In this study, we investigated the role of TLR4 in the development of experimental colitis with regard to the biological actions of macrophage migration inhibitory factor (MIF) using TLR4 null ((-/-)) mice. TLR4(-/-) mice were given 2% dextran sulphate sodium (DSS) in drinking water to induce colitis, which was clinically and histologically as severe as that seen in wild-type (WT) mice. The level of tumour necrosis factor (TNF)-alpha in colon tissues was increased in WT mice but unchanged in TLR4(-/-) mice. The level of myeloperoxidase (MPO) activity in colon tissues was increased by DSS administration in both TLR4(-/-) and WT mice. The expression of MIF was up regulated in the colons of TLR4(-/-) mice with acute DSS-induced colitis. An anti MIF antibody significantly suppressed colitis and elevation of matrix metalloproteinase-13 in TLR4(-/-) mice. The current results obtained from TLR4(-/ ) mice provide evidence that MIF plays a critical role in the development of acute DSS-induced colitis. PMID- 16045731 TI - Interleukin-10 in murine metal-induced systemic autoimmunity. AB - Systemic autoimmune diseases have a complicated and largely unknown aetiology and pathogenesis, but they are at least partly obeying the rules of an ordinary immune response. Cytokines are therefore important in the pathogenesis as demonstrated by the recent success in treating rheumatoid arthritis with anti cytokine agents. The suppressive functions in the immune system have lately received much interest. One of the cytokines in focus in this respect is interleukin (IL)-10. We recently observed that in heavy-metal induced systemic autoimmunity, genetically resistant mice show a strong increase in IL-10 mRNA expression, which was not seen in susceptible mice. We have therefore examined the possible regulating effect of IL-10 on the induction and manifestation of systemic autoimmunity in this model. We took two approaches: a targeted mutation of the IL-10 gene in a strain resistant to heavy metal-induced autoimmunity, and treatment with recombinant IL-10 in the genetically susceptible A.SW strain during the induction of autoimmunity by metals. The wild-type C57BL/6 J (B6-WT) strain did not react with lymphoproliferation, polyclonal B cell activation, anti nucleoar autoantibodies (ANoA) or tissue immune-complex (IC) deposits in response to inorganic mercury (Hg) or silver (Ag). However, serum IgG1 and IgE showed a modest increase during Hg treatment, while Ag caused a weak increase in IgE and IgG2a. The B6.129P2-Il10(tm1Cgn)/J strain (IL-10-deficient mice) did not develop antinucleolar antibodies (ANoA) during Hg treatment, but showed a higher median titre of homogeneous ANA compared with Hg-treated B6-WT mice. Both control and Hg treated (but not Ag-treated) IL-10-deficient mice showed an increase in splenic weight and serum IgG1 compared with B6-WT control and Hg-treated mice. An early, significant increase in serum IgE was seen in Hg-treated IL-10-deficient and WT mice compared with the controls; the increase was 42- and sixfold, respectively. During ongoing intense treatment with rIL-10 in combination with Hg the susceptible A.SW mice showed a reduced development of ANoA and antichromatin antibodies, as well as serum IgE, compared with mice receiving Hg but not rIL-10. In conclusion, IL-10 suppresses several aspects of HgIA, but is not crucial for resistance to heavy metal-induced autoimmunity. Peroral silver treatment suppresses the spontaneous immune activation seen in IL-10-deficient mice. PMID- 16045732 TI - Exposure to Mycobacterium avium induces low-level protection from Mycobacterium bovis infection but compromises diagnosis of disease in cattle. AB - We assessed the effect of exposure to Mycobacterium avium on the development of immune responses and the pathogenesis of disease observed following Mycobacterium bovis challenge. A degree of protection against M. bovis was observed in calves which were pre-exposed to M. avium as assessed by the extent of lesions and bacterial load compared to the M. bovis alone group. The immune response following M. bovis challenge in cattle previously inoculated with M. avium was biased towards antigens (PPD) present in M. avium, whereas the response following M. bovis alone was biased towards antigens present in M. bovis, indicating an imprinting of memory to avian antigens on T lymphocytes. A consequence of the memory to M. avium antigens was failure to diagnose M. bovis infection by the skin test or the IFN(gamma) assay in some of the animals which had lesions of tuberculosis at necropsy. The use of M. bovis specific antigens ESAT-6 and CFP-10 increased IFN(gamma) test specificity in animals previously exposed to M. avium but the responses to these antigens were lower than those observed in animals exposed to M. bovis alone. The implication is that responses to M. avium, although providing some immunity, may mask diagnosis of M. bovis infection, even when specific antigens are employed, potentially contributing to disease transmission in the field. PMID- 16045733 TI - Ex vivo stimulation of cord blood mononuclear cells by dexamethasone and interleukin-7 results in the maturation of interferon-gamma-secreting effector memory T cells. AB - The effects of dexamethasone phosphate and interleukin-7 upon the proliferation of T-cells and the production of interferon-gamma in the newborn's cord blood mononuclear cell cultures were studied. The capability of dexamethasone to enhance T-cell proliferation induced by anti-CD3 with interleukin-7 in some newborn cord blood mononuclear cell cultures was identified. Dexamethasone suppressed production of interferon-gamma in 68-h cell cultures stimulated with anti-CD3 both in the presence of interleukin-7 and without it. However, a 68-h cultivation of newborn blood cells with dexamethasone, anti-CD3 and interleukin-7 resulted in the accumulation of T-lymphocytes capable of producing interferon gamma after restimulation. As a result of it the amount of interferon-gamma producing CD7(+) T-cells and the concentration of interferon-gamma in cultural supernatants were maximal in the cell cultures incubated with anti-CD3, interleukin-7 and dexamethasone during the first 68 h and subsequently restimulated with phorbol 12-myristate 13-acetate and ionomycin. The stimulation of neonatal or adult blood cells by dexamethasone, anti-CD3 and interleukine-7 also causes a decrease in the number of naive T-cells and central memory cells and an increase in the number of effector memory CD7(+)CD45RA(+)CD62L(-) cells in cultures. It is possible that these effects are caused by the influence of dexamethasone on IL-7 receptor expression: it is known that IL-7 receptor alpha chain gene is a glucocorticoid-inducible gene. PMID- 16045734 TI - HUVECs from newborns with a strong family history of myocardial infarction overexpress adhesion molecules and react abnormally to stimulating agents. AB - Atherosclerosis is a complex disease involved in major fatal events such as myocardial infarction and stroke. It is the result of interactions between metabolic, dietetic and environmental risk factors acting on a genetic background that could result in endothelial susceptibility. Our aim was to determine the patterns of expression of adhesion molecules and whether phosphatidylserine is translocated to the cell surface of human umbilical vein endothelial cells (HUVECs) isolated from healthy newborns born to parents with a strong family history of myocardial infarction under TNF-alpha or oxLDL stimulated conditions. Compared to control HUVECs, experimental cords showed: (a) a four-fold increase in VCAM-1 expression under basal conditions, which showed no change after stimulation with the pro-atherogenic factors; (b) a two-fold increase in basal P selectin expression that reached a 10-fold increase with any of the pro atherogenic factors; (c) a basal ICAM-1 expression similar to P-selectin that was not modified by the pro-atherogenic molecules; (d) a similar PECAM-1 expression. Unexpectedly, phospathidylserine expression in experimental cord HUVECs was significantly increased (211 817 versus 3354 TFU) but was not associated to apoptotic death as the percentage of dead cells induced by TNF-alpha treatment was very low (0.55 versus 9.87% in control HUVECs). The latter result was corroborated by TUNEL staining. T cell adherence to HUVECs was highly up regulated in the genetically predisposed samples. The analysis of nonpooled HUVECs, from newborns to family predisposed myocardial-infarction individuals, might represent a useful strategy to identify phenotypical and functional alterations, and hopefully, to take early preventive actions. PMID- 16045735 TI - Interleukin-4 and interleukin-13 enhance CCL26 production in a human keratinocyte cell line, HaCaT cells. AB - Eotaxin-2/CCL24 and eotaxin-3/CCL26 are CC chemokines and their receptor, CC chemokine receptor 3 is preferentially expressed on eosinophils. It was reported that vascular endothelial cells and dermal fibroblasts produced CCL26. However, the regulation of CCL24 and CCL26 production in keratinocytes has not been well documented. We investigated the expression and production of CCL24 and CCL26 in the human keratinocyte cell line, HaCaT cells. Reverse transcription and polymerase chain reaction was performed using these cells and Enzyme-linked immunosorbent assay was carried out using supernatant of these cells. The production of CCL24 in HaCaT cells was slightly enhanced by IL-4 and that of CCL26 was strongly enhanced by IL-4 and IL-13. Furthermore, TNF-alpha generated a synergistic effect on IL-4 enhanced CCL26 production. Dexamethasone, IFN-gamma and the p38 mitogen-activated protein kinase inhibitor SB202190 inhibited IL-4 enhanced CCL26 production. IL-4 enhanced production of CCL26 was inhibited by leflunomide and JAK inhibitor 1, but not by JAK3 inhibitor, which indicates that it is mediated by JAK1-STAT6-dependent pathway. This result also strongly suggests the involvement of the type 2 IL-4 receptor in IL-4 enhanced production of CCL26. These results suggest that keratinocytes are involved in the migration of CC chemokine receptor 3 positive cells such as eosinophils in a Th2-dominant situation like atopic dermatitis. PMID- 16045736 TI - CXCL12 and CXCR4 expression by human gingival fibroblasts in periodontal disease. AB - CXCL12 is a CXC chemokine that is related to lymphocyte infiltration and angiogenesis in inflammatory sites such as arthritis. However, the expression and roles of CXCL12 in periodontal disease are uncertain. The aim of this study was to assess the expression of CXCL12 and its receptor, CXCR4, in periodontal tissue and to investigate the properties of CXCL12 and CXCR4 expression by human gingival fibroblasts (HGF). RT-PCR analysis revealed that CXCL12 and CXCR4 mRNA were expressed in both normal gingival tissues and periodontal diseased tissues. Immunohistochemistry disclosed that CXCL12 was expressed and CXCR4 positive cells were found in both normal and periodontal diseased gingival tissues. Our in vitro experiments elucidated that HGF constitutively produced CXCL12, and the levels were enhanced by stimulation with tumour necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), transforming growth factor-beta (TGF-beta), regulated upon activation normal T cell expressed and secreted (RANTES) and macrophage inflammatory protein 3(alpha) (MIP-3(alpha)). On the other hand, heat killed Porphyromonas gingivalis (P. gingivalis) and P. gingivalis LPS reduced the CXCL12 production by HGF. Flow cytometry analysis clarified that CXCR4 was highly expressed on HGF, and CXCR4 expression was abrogated by TNF-alpha, IFN-gamma and P. gingivalis LPS. Moreover, CXCL12 induced vascular endothelial growth factor (VEGF) production by HGF. Our results demonstrated that CXCL12 might be related to CXCR4+ cells infiltration and angiogenesis both in normal periodontal tissues and periodontal diseased tissue. P. gingivalis, a known periodontal pathogen, inhibits the production of CXCL12 and the expression of CXCR4 by HGF. This fact means that P. gingivalis may inhibit CXCR4+ cells infiltration and neovascularization in periodontal tissue and escape from the immune response. PMID- 16045737 TI - Toxoplasma gondii regulates recruitment and migration of human dendritic cells via different soluble secreted factors. AB - We investigated in vitro the properties of soluble factors produced by Toxoplasma gondii on the recruitment, maturation and migration of human dendritic cells (DC) derived from CD34+ progenitor cells. We used soluble factors including excreted secreted antigens (ESA) produced under various conditions by the virulent type I RH strain (ESA-RH) and the less virulent PRU type II strain (ESA-PRU). Soluble factors of both T. gondii strains appeared to possess a chemokine-like activity that attracted immature DC. This recruitment activity required the presence of functional CCR5 molecules on the cell membrane. Incubation of DC for 24 h with ESA triggered the migration of a large percentage of these cells towards the chemokine MIP-3beta; ESA-PRU was more efficient than ESA-RH. ESA produced in absence of exogenous protein and crude extract did not induce DC migration but retained recruitment activity. These data indicate that recruitment activity and migration-inducing activity are not governed by the same factors. Moreover, incubation of DC for 48 h with ESA did not modify the expression of costimulation or maturation markers (CD83, CD40, CD80, CD86 or HLA-DR), but induced a decrease in CCR6 expression associated with an increased expression of CCR7. Taken together, these results suggest that T. gondii controls recruitment and migration of immature DC by different soluble factors and may induce a dysfunction in the host-specific immune response. PMID- 16045738 TI - CDC staging based on absolute CD4 count and CD4 percentage in an HIV-1-infected Indian population: treatment implications. AB - CD4+ T-cell levels are an important criterion for categorizing HIV-related clinical conditions according to the CDC classification system and are therefore important in the management of HIV by initiating antiretroviral therapy and prophylaxis for opportunistic infections due to HIV among HIV-infected individuals. However, it has been observed that the CD4 counts are affected by the geographical location, race, ethnic origin, age, gender and changes in total and differential leucocyte counts. In the light of this knowledge, we classified 600 HIV seropositive antiretroviral treatment (ART)-naive Indian individuals belonging to different CDC groups A, B and C on the basis of CDC criteria of both CD4% and CD4 counts and receiver operating characteristic (ROC) curves were generated. Importantly, CDC staging on the basis of CD4% indicated significant clinical implications, requiring an early implementation of effective antiretroviral treatment regimen in HIV-infected individuals deprived of treatment when classified on the basis of CD4 counts. PMID- 16045739 TI - Circulating anti-actin and anti-ATP synthase antibodies identify a sub-set of patients with idiopathic nephrotic syndrome. AB - Idiopathic nephrotic syndrome (iNS) with resistance or dependence to steroids is a common disease in children but in spite of an increasing clinical impact its pathogenesis is unknown. We screened for the presence of circulating antibodies against glomerular (podocytes, mesangium) and tubular cells (tubular epithelia) a cohort of 60 children with iNS including 8 patients with a familial trait of iNS or with proven mutation of NPHS1-NPHS2 and 12 with good sensitivity to steroids. Positive sera were found in 8 cases, all belonging to the category without familial trait/molecular defects. The targets of antibodies were characterized with Western blot and MALDI-Mass utilizing beta-hexyl cell extracts separated with two-dimensional electrophoresis. In all cases antibodies of the IgM class were directed against ATP synthase beta chain alone (4 cases) or in combination with actin (3 cases); one child presented IgG against aldose reductase. The clinical picture was nephrotic syndrome with steroid resistance or dependence and variable cyclosporin sensitivity; 3 patients developed end stage renal failure. The basic pathology picture was focal segmental glomerulosclerosis (FSGS) in 4 cases and mesangial proliferative glomerulonephrites with deposition of IgM in 2. Overall, patients with circulating auto-antibodies could not be readely differentiated on clinical grounds with the exception of 3 children who developed positivity for antinuclear antibodies during the follow-up. Affinity-purified IgM from one patient who underwent plasmapheresis for therapeutical pourposes (but not from a normal pool) induced proteinuria in Sprague-Dawley rats and concomitant human IgM deposition within glomeruli. This is the first report of circulating anti-actin/ATP synthase beta chain antibodies in a subset of patients with iNS. Both pathological significance and clinical impact given by the presence of these antibodies and the relationship with other conditions such as lupus-erythematosus, characterized by their presence, must be defined. PMID- 16045740 TI - Antibody to severe acute respiratory syndrome (SARS)-associated coronavirus spike protein domain 2 cross-reacts with lung epithelial cells and causes cytotoxicity. AB - Both viral effect and immune-mediated mechanism are involved in the pathogenesis of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) infection. In this study, we showed that in SARS patient sera there were autoantibodies (autoAbs) that reacted with A549 cells, the type-2 pneumocytes, and that these autoAbs were mainly IgG. The autoAbs were detectable 20 days after fever onset. Tests of non-SARS-pneumonia patients did not show the same autoAb production as in SARS patients. After sera IgG bound to A549 cells, cytotoxicity was induced. Cell cytotoxicity and the anti-epithelial cell IgG level were positively correlated. Preabsorption and binding assays indicated the existence of cross reactive epitopes on SARS-CoV spike protein domain 2 (S2). Furthermore, treatment of A549 cells with anti-S2 Abs and IFN-gamma resulted in an increase in the adherence of human peripheral blood mononuclear cells to these epithelial cells. Taken together, we have demonstrated that the anti-S2 Abs in SARS patient sera cause cytotoxic injury as well as enhance immune cell adhesion to epithelial cells. The onset of autoimmune responses in SARS-CoV infection may be implicated in SARS pathogenesis. PMID- 16045741 TI - An increased MRP8/14 expression and adhesion, but a decreased migration towards proinflammatory chemokines of type 1 diabetes monocytes. AB - In the early development of type 1 diabetes macrophages and dendritic cells accumulate around the islets of Langerhans at sites of fibronectin expression. It is thought that these macrophages and dendritic cells are derived from blood monocytes. Previously, we showed an increased serum level of MRP8/14 in type 1 diabetes patients that induced healthy monocytes to adhere more strongly to fibronectin (FN). Here we show that MRP8/14 is expressed and produced at a higher level by type 1 diabetes monocytes, particularly after adhesion to FN, creating a positive feedback mechanism for a high fibronectin-adhesive capacity. Also adhesion to endothelial cells was increased in type 1 diabetes monocytes. Despite this increased adhesion the transendothelial migration of monocytes of type 1 diabetes patients was decreased towards the proinflammatory chemokines CCL2 and CCL3. Because non-obese diabetic (NOD) mouse monocytes show a similar defective proinflammatory migration, we argue that an impaired monocyte migration towards proinflammatory chemokines might be a hallmark of autoimmune diabetes. This hampered monocyte response to proinflammatory chemokines questions whether the early macrophage and dendritic cell accumulation in the diabetic pancreas originates from an inflammatory-driven influx of monocytes. We also show that the migration of type 1 diabetes monocytes towards the lymphoid tissue-related CCL19 was increased and correlated with an increased CCR7 surface expression on the monocytes. Because NOD mice show a high expression of these lymphoid tissue related chemokines in the early pancreas it is more likely that the early macrophage and dendritic cell accumulation in the diabetic pancreas is related to an aberrant high expression of lymphoid tissue-related chemokines in the pancreas. PMID- 16045742 TI - Sam68 is tyrosine phosphorylated and recruited to signalling in peripheral blood mononuclear cells from HIV infected patients. AB - Human immunodeficiency virus (HIV) codes for a protein, Rev, that mediates the viral RNA export from the nucleus to the cytoplasm. Recently, it has been found that Sam68, the substrate of Src associated in mitosis, is a functional homologue of Rev, and a synergistic activator of Rev activity. Thus, it has been suggested that Sam68 may play an important role in the post-transcriptional regulation of HIV. Sam68 contains an RNA binding motif named KH [homology to the nuclear ribonucleoprotein (hnRNP) K]. Tyrosine phosphorylation of Sam68 and binding to SH3 domains have been found to negatively regulate its RNA binding capacity. Besides, tyrosine phosphorylation of Sam68 allows the formation of signalling complexes with other proteins containing SH2 and SH3 domains, suggesting a role in signal transduction of different systems in human lymphocytes, such as the T cell receptor, and leptin receptor, or the insulin receptor in other cell types. In the present work, we have found that Sam68 is tyrosine phosphorylated in peripheral blood mononuclear cells (PBMC) from HIV infected subjects, leading to the formation of signalling complexes with p85 the regulatory subunit of PI3K, GAP and STAT-3, and decreasing its RNA binding capacity. In contrast, PBMC from HIV infected subjects have lower expression levels of Sam68 compared with controls. These results suggest that Sam68 may play some role in the immune function of lymphocytes in HIV infection. PMID- 16045743 TI - 2B4 expression on natural killer cells increases in HIV-1 infected patients followed prospectively during highly active antiretroviral therapy. AB - Human immunodeficiency virus (HIV)-1 infection influences natural killer (NK) cell expression of inhibitory NK receptors and activating natural cytotoxicity receptors. It is unknown whether expression of the co-stimulatory NK cell receptor 2B4 (CD244) on NK cells and CD3+ CD8+ cells are affected by highly active antiretroviral therapy (HAART), low-level viraemia, proviral-DNA or immune activation in HIV-1 infected patients. A total of 101 HAART-treated HIV-1 infected patients with < or = 200 HIV-RNA copies/ml were followed prospectively for 24 months. HIV-RNA was investigated 3-monthly and 2B4 expression on CD3- CD16+ NK cells and CD3+ CD8+ cells, proviral-DNA and plasma soluble tumour necrosis factor receptor (sTNFr)-II were investigated 6-monthly. For comparison, 2B4 expression was investigated in 20 healthy individuals. The concentration of 2B4+ NK cells was initially reduced in HIV-1 infected patients (P < 0.001) but increased to a normal level during the 24 months' follow-up. The concentration of CD3+ CD8+ 2B4+ cells in HIV-1 infected patients was normal and did not change during follow-up. The relative fluorescence intensity (RFI) of 2B4 increased on both NK cells and CD3+ CD8+ cells during follow-up (both P < 0.001). Higher levels of proviral-DNA carrying cells and plasma sTNFrII were associated with reductions in the concentration of 2B4+ NK cells (all P < 0.05). HIV-RNA had no effect on 2B4 expression on NK cells or CD3+ CD8+ cells. These findings demonstrate that the concentration of 2B4+ NK cells normalizes during long-term HAART in HIV-1 infected patients. The finding that proviral-DNA and sTNFrII were associated negatively with the concentration of 2B4+ NK cells suggests that immune activation in HIV-1 infected patients receiving HAART influences the target cell recognition by NK cells. PMID- 16045744 TI - The direct and indirect allogeneic presentation pathway during acute rejection after human cardiac transplantation. AB - Alloreactive T cells may be activated via a direct or an indirect antigen presentation pathway. We questioned whether the frequency of interferon (IFN) gamma producing cells determined by enzyme-linked immunospot (ELISPOT) assay is an effective tool to monitor the direct and/or indirect presentation pathway. Secondly, we wondered whether early and late acute rejection (AR) are associated with both pathways. Before (n = 15), during (n = 18) and after (n = 16) a period of AR, peripheral blood mononuclear cell (PBMC) samples were tested from 13 heart transplant recipients. The direct presentation pathway was always present. The number of IFN-gamma producing cells reactive to this pathway increased significantly (P = 0.04) during AR and the number decreased (P = 0.005) after AR therapy. In contrast, the indirect allogeneic presentation pathway was present in only eight of 18 AR samples. When the indirect presentation pathway was detectable, it increased significantly during AR. Five of eight of these AR occurred more than 6 months after transplantation. The ELISPOT assay, enumerating alloreactive IFN-gamma producing cells, is a valuable tool to determine the reactivity via both the direct and the indirect presentation pathway. The direct presentation pathway always plays a role in AR, while the indirect pathway contributes especially to late AR. PMID- 16045745 TI - Detection of in vitro interferon-gamma and serum tumour necrosis factor-alpha in multidrug-resistant tuberculosis patients. AB - Multidrug-resistant tuberculosis (MDR-TB) is known as having a poor prognosis with a weak response to therapy and very high death rates. The aim of this work was to assess the immune response to the RD1-encoded antigen ESAT-6 of Mycobacterium tuberculosis in MDR-TB patients and compare to non-resistant (NR) TB patients and healthy controls (HC). Evaluation of interferon (IFN)-gamma production showed that, although 55% of the MDR patients were responsive to ESAT 6, they produced lower IFN-gamma levels (553 +/- 11 pg/ml) when compared to NR-TB (1179 +/- 163 pg/ml; P < 0.05) but not to controls (412 +/- 65.7 pg/ml). Differences in the response to ESAT-6 and to its overlapping peptides mixture were also significant between MDR versus treated pulmonary NR-TB. Furthermore, a very low rate of response to PPD (23.5%) and to Ag85B (33.3%) was noted in MDR-TB patients as compared to the other groups. To determine the inflammatory response in patients' groups, detection of tumour necrosis factor (TNF)-alpha was assessed in their sera before and during chemotherapy. Mean TNF-alpha levels in MDR-TB (43.8 +/- 9 pg/ml) paralleled those found in treated pulmonary, and it was significantly different (P < 0.05) from the values found in untreated NR and HC. Interestingly, secretion of IFN-gamma and TNF-alpha were predominant in MDR patients who presented with bilateral pulmonary lesions and lung cavitation. The present data indicate that the overall immune response to mycobacterial antigens is decreased in resistant TB and the major role inflammatory cytokines may play in perpetuating pulmonary tissue damage. PMID- 16045746 TI - FoxP3(+)CD4(+)CD25(+) T cells with regulatory properties can be cultured from colonic mucosa of patients with Crohn's disease. AB - Summary CD4(+)CD25(+) regulatory T cells (T(regs)) are involved in the maintenance of peripheral tolerance and ensure a balanced immune response competent of fighting pathogens and at the same time recognizing commensals as harmless. This feature is lost in Crohn's disease (CD). The forkhead/winged helix transcription factor FoxP3 is a master gene for T(reg) function and defects in the FoxP3 gene lead to a clinical picture similar to inflammatory bowel disease (IBD). Murine colitis can be cured by adoptive transfer of T(regs) and ex vivo generated gut-specific T(regs) represent an attractive option for therapy in CD. Thus, defective T(regs) could contribute to the development of CD. We cultured biopsies of colonic mucosa in the presence of high concentrations of interleukin (IL)-2 and IL-4 to overcome the anergic nature of naturally occurring CD4(+)CD25(+) T(regs) in the mucosa. We investigated the expression of FoxP3 and regulatory potential of gut-derived CD4(+)CD25(+) T cells cultured from patients with CD and healthy individuals. The FoxP3 expression was analysed by reverse transcriptase polymerase chain reaction (RT-PCR), and the suppressive effect of FoxP3(+)CD4(+)CD25(+) T cells on proliferation and cytokine production of autologous CD4(+) T cells was assessed by flow cytometry. Cultured gut-derived T cells with CD4(+)CD25(+) phenotype expressed FoxP3 and were able as the freshly isolated T(regs) from peripheral blood to suppress proliferation and cytokine production of autologous CD4(+) T cells. Thus, we demonstrate that FoxP3(+)CD4(+)CD25(+) T cells with regulatory properties can be propagated in vitro from inflamed mucosa of CD patients, which may be of interest in adoptive immunotherapy. PMID- 16045747 TI - Rare naturally occurring immune responses to three epitopes from the widely expressed tumour antigens hTERT and CYP1B1 in multiple myeloma patients. AB - The widely expressed tumour antigens hTERT and CYP1B1 are commonly expressed in multiple myeloma (MM) cells. Several trials targeting these antigens by immunotherapy have been initiated. The aim of this study was to explore whether patients with MM have an endogenous pre-existing immune response against recently identified epitopes from hTERT and CYP1B1. Peripheral blood T cells from 27 HLA A*0201+ multiple myeloma patients at different stages of disease and 20 healthy HLA-A*0201+ donors were enriched and studied for the presence of hTERT- and CYP1B1-specific cytotoxic T cells using MHC tetramer detection and short-term ex vivo expansion. No significant expansion of tetramer-positive cells was detected in the peripheral blood of either MM patients or healthy controls when cells were stained with tetramers containing the dominant hTERT-derived epitope or two peptides derived from CYP1B1. A single ex vivo peptide stimulation led to the detection of a small population (0.3-0.5%) of hTERT-specific cells in two of 27 patients with MM. None of the patients or controls showed significant expansion of CYP1B1-specific cells after a single peptide stimulation. Thus, endogenous in vivo priming of T cells against hTERT and CYP1B1 is a rare event in MM patients. These results suggest that strategies targeting hTERT and CYP1B1 may have to utilize techniques to induce T cell responses from a naive precursor frequency. PMID- 16045748 TI - Origins of DNA replication in the three domains of life. AB - Replication of DNA is essential for the propagation of life. It is somewhat surprising then that, despite the vital nature of this process, cellular organisms show a great deal of variety in the mechanisms that they employ to ensure appropriate genome duplication. This diversity is manifested along classical evolutionary lines, with distinct combinations of replicon architecture and replication proteins being found in the three domains of life: the Bacteria, the Eukarya and the Archaea. Furthermore, although there are mechanistic parallels, even within a given domain of life, the way origins of replication are defined shows remarkable variation. PMID- 16045749 TI - BCR kinase phosphorylates 14-3-3 Tau on residue 233. AB - The breakpoint cluster region protein, BCR, has protein kinase activity that can auto- and trans-phosphorylate serine, threonine and tyrosine residues. BCR has been implicated in chronic myelogenous leukaemia as well as important signalling pathways, and as such its interaction with 14-3-3 is of major interest. 14-3-3tau and zeta isoforms have been shown previously to be phosphorylated in vitro and in vivo by BCR kinase on serine and threonine residue(s) but site(s) were not determined. Phosphorylation of 14-3-3 isoforms at distinct sites is an important mode of regulation that negatively affects interaction with Raf kinase and Bax, and potentially influences the dimerization of 14-3-3. In this study we have further characterized the BCR-14-3-3 interaction and have identified the site phosphorylated by BCR. We show here that BCR interacts with at least five isoforms of 14-3-3 in vivo and phosphorylates 14-3-3tau on Ser233 and to a lesser extent 14-3-3zeta on Thr233. We have previously shown that these two isoforms are also phosphorylated at this site by casein kinase 1, which, in contrast to BCR, preferentially phosphorylates 14-3-3zeta. PMID- 16045750 TI - Nuclear aggregates of polyamines are supramolecular structures that play a crucial role in genomic DNA protection and conformation. AB - In a previous study we showed that natural polyamines interact in the nuclear environment with phosphate groups to form molecular aggregates [nuclear aggregates of polyamines (NAPs)] with estimated molecular mass values of 8000, 4800 and 1000 Da. NAPs were found to interact with genomic DNA, influence its conformation and interfere with the action of nucleases. In the present work, we demonstrated that NAPs protect naked genomic DNA from DNase I, whereas natural polyamines (spermine, spermidine and putrescine) fail to do so. In the context of DNA protection, NAPs induced noticeable changes in DNA conformation, which were revealed by temperature-dependent modifications of DNA electrophoretic properties. In addition, we presented, for NAPs, a structural model of polyamine aggregation into macropolycyclic compounds. We believe that NAPs are the sole biological forms by which polyamines efficiently protect genomic DNA against DNase I, while maintaining its dynamic structure. PMID- 16045751 TI - Functional interaction between RNA helicase II/Gu(alpha) and ribosomal protein L4. AB - RNA helicase II/Gu(alpha) is a multifunctional nucleolar protein involved in ribosomal RNA processing in Xenopus laevis oocytes and mammalian cells. Downregulation of Gu(alpha) using small interfering RNA (siRNA) in HeLa cells resulted in 80% inhibition of both 18S and 28S rRNA production. The mechanisms underlying this effect remain unclear. Here we show that in mammalian cells, Gu(alpha) physically interacts with ribosomal protein L4 (RPL4), a component of 60S ribosome large subunit. The ATPase activity of Gu(alpha) is important for this interaction and is also necessary for the function of Gu(alpha) in the production of both 18S and 28S rRNAs. Knocking down RPL4 expression using siRNA in mouse LAP3 cells inhibits the production of 47/45S, 32S, 28S, and 18S rRNAs. This inhibition is reversed by exogenous expression of wild-type human RPL4 protein but not the mutant form lacking Gu(alpha)-interacting motif. These observations have suggested that the function of Gu(alpha) in rRNA processing is at least partially dependent on its ability to interact with RPL4. PMID- 16045752 TI - Structural analysis of the N-glycans of the major cysteine proteinase of Trypanosoma cruzi. Identification of sulfated high-mannose type oligosaccharides. AB - Trypanosoma cruzi, the parasitic protozoan that causes Chagas disease, contains a major cysteine proteinase, cruzipain. This lysosomal enzyme bears an unusual C terminal extension that contains a number of post-translational modifications, and most antibodies in natural and experimental infections are directed against it. In this report we took advantage of UV-MALDI-TOF mass spectrometry in conjunction with peptide N-glycosidase F deglycosylation and high performance anion exchange chromatography analysis to address the structure of the N-linked oligosaccharides present in this domain. The UV-MALDI-TOF MS analysis in the negative-ion mode, using nor-harmane as matrix, allowed us to determine a new striking feature in cruzipain: sulfated high-mannose type oligosaccharides. Sulfated GlcNAc2Man3 to GlcNAc2Man9 species were identified. In accordance, after chemical or enzymatic desulfation, the corresponding signals disappeared. In addition, by UV-MALDI-TOF MS analysis (a) a main population of high-mannose type oligosaccharides was shown in the positive-ion mode, (b) lactosaminic glycans were also identified, among them, structures corresponding to monosialylated species were detected, and (c) as an interesting fact a fucosylated oligosaccharide was also detected. The presence of the deoxy sugar was further confirmed by high performance anion exchange chromatography. In conclusion, the total number of oligosaccharides occurring in cruzipain was shown to be much higher than previous estimates. This constitutes the first report on the presence of sulfated glycoproteins in Trypanosomatids. PMID- 16045753 TI - Mechanistic investigation of a highly active phosphite dehydrogenase mutant and its application for NADPH regeneration. AB - NAD(P)H regeneration is important for biocatalytic reactions that require these costly cofactors. A mutant phosphite dehydrogenase (PTDH-E175A/A176R) that utilizes both NAD and NADP efficiently is a very promising system for NAD(P)H regeneration. In this work, both the kinetic mechanism and practical application of PTDH-E175A/A176R were investigated for better understanding of the enzyme and to provide a basis for future optimization. Kinetic isotope effect studies with PTDH-E175A/A176R showed that the hydride transfer step is (partially) rate determining with both NAD and NADP giving (D)V values of 2.2 and 1.7, respectively, and (D)V/K(m,phosphite) values of 1.9 and 1.7, respectively. To better comprehend the relaxed cofactor specificity, the cofactor dissociation constants were determined utilizing tryptophan intrinsic fluorescence quenching. The dissociation constants of NAD and NADP with PTDH-E175A/A176R were 53 and 1.9 microm, respectively, while those of the products NADH and NADPH were 17.4 and 1.22 microm, respectively. Using sulfite as a substrate mimic, the binding order was established, with the cofactor binding first and sulfite binding second. The low dissociation constant for the cofactor product NADPH combined with the reduced values for (D)V and k(cat) implies that product release may become partially rate determining. However, product inhibition does not prevent efficient in situ NADPH regeneration by PTDH-E175A/A176R in a model system in which xylose was converted into xylitol by NADP-dependent xylose reductase. The in situ regeneration proceeded at a rate approximately fourfold faster with PTDH E175A/A176R than with either WT PTDH or a NADP-specific Pseudomonas sp.101 formate dehydrogenase mutant with a total turnover number for NADPH of 2500. PMID- 16045754 TI - Identification of a novel thyroid hormone-sulfating cytosolic sulfotransferase, SULT1 ST5, from zebrafish. AB - By employing RT-PCR in conjunction with 3'-RACE, a full-length cDNA encoding a novel zebrafish cytosolic sulfotransferase (SULT) was cloned and sequenced. Sequence analysis revealed that this zebrafish SULT (designated SULT1 ST5) is, at the amino acid sequence level, close to 50% identical to human and dog SULT1B1 (thyroid hormone SULT). A recombinant form of zebrafish SULT1 ST5 was expressed using the pGEX-2TK bacterial expression system and purified from transformed BL21 (DE3) cells. Purified zebrafish SULT1 ST5 migrated as a 34 kDa protein and displayed substrate specificity for thyroid hormones and their metabolites among various endogenous compounds tested. The enzyme also exhibited sulfating activities toward some xenobiotic phenolic compounds. Its pH optima were 6.0 and 9.0 with 3,3',5-triiodo-l-thyronine (l-T3) as substrate and 6.0 with beta naphthol as substrate. Kinetic constants of the enzyme with thyroid hormones and their metabolites as substrates were determined. Quantitative evaluation of the regulatory effects of divalent metal cations on the l-T3-sulfating activity of SULT1 ST5 revealed that Fe2+, Hg2+, Co2+, Zn2+, Cu2+, Cd2+ and Pb2+ exhibited dramatic inhibitory effects, whereas Mn2+ showed a significant stimulation. Developmental stage-dependent expression experiments revealed a significant level of expression of this novel zebrafish thyroid hormone-sulfating SULT at the beginning of the hatching period during embryogenesis, which gradually increased to a high level of expression throughout the larval stage into maturity. PMID- 16045755 TI - Selenium affects biosilica formation in the demosponge Suberites domuncula. Effect on gene expression and spicule formation. AB - Selenium is a trace element found in freshwater and the marine environment. We show that it plays a major role in spicule formation in the demosponge Suberites domuncula. If added to primmorphs, an in vitro sponge cell culture system, it stimulates the formation of siliceous spicules. Using differential display of transcripts, we demonstrate that, after a 72-h exposure of primmorphs to selenium, two genes are up-regulated; one codes for selenoprotein M and the other for a novel spicule-associated protein. The deduced protein sequence of selenoprotein M (14 kDa) shows characteristic features of metazoan selenoproteins. The spicule-associated protein (26 kDa) comprises six characteristic repeats of 20 amino acids, composed of 10 distinct hydrophobic regions ( approximately 9 amino acids in length). Recombinant proteins were prepared, and antibodies were raised against these two proteins. Both were found to stain the central axial filament, which comprises the silicatein, as well as the surface of the spicules. In the presence of selenium, only the genes for selenoprotein M and spicule-associated protein are up-regulated, whereas the expression of the silicatein gene remains unchanged. Finally we show that, in the presence of selenium, larger silica aggregates are formed. We conclude that selenium has a stimulatory effect on the formation of siliceous spicules in sponges, and it may be involved in the enzymatic synthesis of biosilica components. PMID- 16045756 TI - Myocyte enhancer factor 2 (MEF2) is a key modulator of the expression of the prothoracicotropic hormone gene in the silkworm, Bombyx mori. AB - Prothoracicotropic hormone (PTTH) plays a central role in controlling molting, metamorphosis, and diapause termination in insects by stimulating the prothoracic glands to synthesize and release the molting hormone, ecdysone. Using Autographa californica nucleopolyhedrovirus (AcNPV)-mediated transient gene transfer into the central nervous sytem (CNS) of the silkworm, Bombyx mori, we identified two cis-regulatory elements that participate in the decision and the enhancement of PTTH gene expression in PTTH-producing neurosecretory cells (PTPCs). The cis element mediating the enhancement of PTTH gene expression binds the transcription factor Bombyx myocyte enhancer factor 2 (BmMEF2). The BmMEF2 gene was expressed in various tissues including the CNS. In brain, the BmMEF2 gene was expressed at elevated levels in two types of lateral neurosecretory cells, namely PTPCs and corazonin-like immunoreactive lateral neurosecretory cells. Overexpression of BmMEF2 cDNA caused an increase in the transcription of PTTH. Therefore, BmMEF2 appears to be particularly important in the brain where it is responsible for the differentiation of lateral neurosecretory cells, including the enhancement of PTTH gene expression. This is the first report to identify a target gene of MEF2 in the invertebrate nervous system. PMID- 16045757 TI - An antimicrobial peptide tachyplesin acts as a secondary secretagogue and amplifies lipopolysaccharide-induced hemocyte exocytosis. AB - In the horseshoe crab, bacterial lipopolysaccharide (LPS) induces exocytosis by granular hemocytes, resulting in the secretion of various defense molecules, such as lectins and antimicrobial peptides, via a G protein-mediating signaling pathway. This response is a key component of the horseshoe crab innate immune response against infectious microorganisms. Here, we report an endogenous amplification mechanism for LPS-induced hemocytes exocytosis. The concentration of LPS required for maximal secretion decreased in proportion to the density of hemocytes, suggesting the presence of a positive feedback mechanism for secretion via a mediator secreted from hemocytes. The exocytosed fluid of hemocytes was found able to induce hemocyte exocytosis in the absence of LPS. Furthermore, tachyplesin, a major antimicrobial peptide of hemocytes, was able to trigger exocytosis in an LPS-independent manner, which was inhibited by a phospholipase C inhibitor, U-73122, and a G protein inhibitor, pertussis toxin. Surface plasmon resonance analysis showed that tachyplesin directly interacts with bovine G protein. These findings suggest that the tachyplesin-induced hemocyte exocytosis also occurs via a G protein-mediating signaling pathway. We concluded that tachyplesin functions not only as an antimicrobial substance, but also as a secondary secretagogue of LPS-induced hemocyte exocytosis, leading to the amplification of the innate immune reaction at sites of injury. PMID- 16045758 TI - Template requirements and binding of hepatitis C virus NS5B polymerase during in vitro RNA synthesis from the 3'-end of virus minus-strand RNA. AB - In our attempt to obtain further information on the replication mechanism of the hepatitis C virus (HCV), we have studied the role of sequences at the 3'-end of HCV minus-strand RNA in the initiation of synthesis of the viral genome by viral RNA-dependent RNA polymerase (RdRp). In this report, we investigated the template and binding properties of mutated and deleted RNA fragments of the 3'-end of the minus-strand HCV RNA in the presence of viral polymerase. These mutants were designed following the newly established secondary structure of this viral RNA fragment. We showed that deletion of the 3'-SL-A1 stem loop significantly reduced the level of RNA synthesis whereas modifications performed in the SL-B1 stem loop increased RNA synthesis. Study of the region encompassing the 341 nucleotides of the 3'-end of the minus-strand RNA shows that these two hairpins play a very limited role in binding to the viral polymerase. On the contrary, deletions of sequences in the 5'-end of this fragment greatly impaired both RNA synthesis and RNA binding. Our results strongly suggest that several domains of the 341 nucleotide region of the minus-strand 3'-end interact with HCV RdRp during in vitro RNA synthesis, in particular the region located between nucleotides 219 and 239. PMID- 16045759 TI - Mapping the functional domains of human transcobalamin using monoclonal antibodies. AB - Recombinant human transcobalamin (TC) was probed with 17 monoclonal antibodies (mAbs), using surface plasmon resonance measurements. These experiments identified five distinct epitope clusters on the surface of holo-TC. Western blot analysis of the CNBr cleavage fragments of TC allowed us to distribute the epitopes between two regions, which spanned either the second quarter of the TC sequence GQLA...TAAM(103-198) or the C-terminal peptide LEPA...LVSW(316-427). Proteolytic fragments of TC and the synthetic peptides were used to further specify the epitope map and define the functional domains of TC. Only one antibody showed some interference with cobalamin (Cbl) binding to TC, and the corresponding epitope was situated at the C-terminal stretch TQAS...QLLR(372 399). We explored the receptor-blocking effect of several mAbs and heparin to identify TC domains essential for the interaction between holo-TC and the receptor. The receptor-related epitopes were located within the TC sequence GQLA...HHSV(103-159). The putative heparin-binding site corresponded to a positively charged segment KRSN...RTVR(207-227), which also seemed to be necessary for receptor binding. We conclude that conformational changes in TC upon Cbl binding are accompanied by the convergence of multiple domains, and only the assembled conformation of the protein (i.e. holo-TC) has high affinity for the receptor. PMID- 16045760 TI - Enlarging the gas access channel to the active site renders the regulatory hydrogenase HupUV of Rhodobacter capsulatus O2 sensitive without affecting its transductory activity. AB - In the photosynthetic bacterium Rhodobacter capsulatus, the synthesis of the energy-producing hydrogenase, HupSL, is regulated by the substrate H2, which is detected by a regulatory hydrogenase, HupUV. The HupUV protein exhibits typical features of [NiFe] hydrogenases but, interestingly, is resistant to inactivation by O2. Understanding the O2 resistance of HupUV will help in the design of hydrogenases with high potential for biotechnological applications. To test whether this property results from O2 inaccessibility to the active site, we introduced two mutations in order to enlarge the gas access channel in the HupUV protein. We showed that such mutations (Ile65-->Val and Phe113-->Leu in HupV) rendered HupUV sensitive to O2 inactivation. Also, in contrast with the wild-type protein, the mutated protein exhibited an increase in hydrogenase activity after reductive activation in the presence of reduced methyl viologen (up to 30% of the activity of the wild-type). The H2-sensing HupUV protein is the first component of the H2-transduction cascade, which, together with the two-component system HupT/HupR, regulates HupSL synthesis in response to H2 availability. In vitro, the purified mutant HupUV protein was able to interact with the histidine kinase HupT. In vivo, the mutant protein exhibited the same hydrogenase activity as the wild-type enzyme and was equally able to repress HupSL synthesis in the absence of H2. PMID- 16045761 TI - Consequences of COP9 signalosome and 26S proteasome interaction. AB - The COP9 signalosome (CSN) occurs in all eukaryotic cells. It is a regulatory particle of the ubiquitin (Ub)/26S proteasome system. The eight subunits of the CSN possess sequence homologies with the polypeptides of the 26S proteasome lid complex and just like the lid, the CSN consists of six subunits with PCI (proteasome, COP9 signalosome, initiation factor 3) domains and two components with MPN (Mpr-Pad1-N-terminal) domains. Here we show that the CSN directly interacts with the 26S proteasome and competes with the lid, which has consequences for the peptidase activity of the 26S proteasome in vitro. Flag-CSN2 was permanently expressed in mouse B8 fibroblasts and Flag pull-down experiments revealed the formation of an intact Flag-CSN complex, which is associated with the 26S proteasome. In addition, the Flag pull-downs also precipitated cullins indicating the existence of super-complexes consisting of the CSN, the 26S proteasome and cullin-based Ub ligases. Permanent expression of a chimerical subunit (Flag-CSN2-Rpn6) consisting of the N-terminal 343 amino acids of CSN2 and of the PCI domain of S9/Rpn6, the paralog of CSN2 in the lid complex, did not lead to the assembly of an intact complex showing that the PCI domain of CSN2 is important for complex formation. The consequence of permanent Flag-CSN2 overexpression was de-novo assembly of the CSN complex connected with an accelerated degradation of p53 and stabilization of c-Jun in B8 cells. The possible role of super-complexes composed of the CSN, the 26S proteasome and of Ub ligases in the regulation of protein stability is discussed. PMID- 16045762 TI - Altered deoxyribonucleotide pools in T-lymphoblastoid cells expressing the multisubstrate nucleoside kinase of Drosophila melanogaster. AB - The multisubstrate nucleoside kinase of Drosophila melanogaster (Dm-dNK) can be expressed in human solid tumor cells and its unique enzymatic properties makes this enzyme a suicide gene candidate. In the present study, Dm-dNK was stably expressed in the CCRF-CEM and H9 T-lymphoblastoid cell lines. The expressed enzyme was localized to the cell nucleus and the enzyme retained its activity. The Dm-dNK overexpressing cells showed approximately 200-fold increased sensitivity to the cytostatic activity of several nucleoside analogs, such as the pyrimidine nucleoside analogs (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU) and 1 beta-d-arabinofuranosylthymine (araT), but not to the antiherpetic purine nucleoside analogs ganciclovir, acyclovir and penciclovir, which may allow this technology to be applied in donor T cells and/or rescue graft vs. host disease to permit modulation of alloreactivity after transplantation. The most pronounced effect on the steady-state dNTP levels was a two- to 10-fold increased dTTP pool in Dm-dNK expressing cells that were grown in the presence of 1 microm of each natural deoxyribonucleoside. Although the Dm-dNK expressing cells demonstrated dNTP pool imbalances, no mitochondrial DNA deletions or altered mitochondrial DNA levels were detected in the H9 Dm-dNK expressing cells. PMID- 16045763 TI - Peroxiredoxin II functions as a signal terminator for H2O2-activated phospholipase D1. AB - Phospholipase D1 (PLD1) is a signal-transduction regulated enzyme which regulates several cell intrinsic processes including activation of NAPDH oxidase, which elevates intracellular H2O2. Several proteins have been reported to interact with PLD1 in resting cells. We sought to identify proteins that interact with PLD1 after phorbol 12-myristate 13-acetate (PMA) stimulation. A novel interaction with peroxiredoxin II (PrxII), an enzyme that eliminates cellular H2O2, which is a known stimulator of PLD1, was identified by PLD1-affinity pull-down and MS. PMA stimulation was confirmed to promote physical interaction between PLD1 and PrxII and to cause PLD1 and PrxII to colocalize subcellularly. Functional significance of the interaction was suggested by the observation that over-expression of PrxII specifically reduces the response of PLD1 to stimulation by H2O2. These results indicate that PrxII may have a signal-terminating role for PLD1 by being recruited to sites containing activated PLD1 after cellular stimulation involving production of H2O2. PMID- 16045764 TI - The Alzheimer beta-peptide shows temperature-dependent transitions between left handed 3-helix, beta-strand and random coil secondary structures. AB - The temperature-induced structural transitions of the full length Alzheimer amyloid beta-peptide [A(beta)(1-40) peptide] and fragments of it were studied using CD and 1H NMR spectroscopy. The full length peptide undergoes an overall transition from a state with a prominent population of left-handed 3(1) (polyproline II; PII)-helix at 0 degrees C to a random coil state at 60 degrees C, with an average DeltaH of 6.8 +/- 1.4 kJ.mol(-1) per residue, obtained by fitting a Zimm-Bragg model to the CD data. The transition is noncooperative for the shortest N-terminal fragment A(beta)(1-9) and weakly cooperative for A(beta)(1-40) and the longer fragments. By analysing the temperature-dependent 3J(HNH(alpha)) couplings and hydrodynamic radii obtained by NMR for A(beta)(1-9) and A(beta)(12-28), we found that the structure transition includes more than two states. The N-terminal hydrophilic A(beta)(1-9) populates PII-like conformations at 0 degrees C, then when the temperature increases, conformations with dihedral angles moving towards beta-strand at 20 degrees C, and approaches random coil at 60 degrees C. The residues in the central hydrophobic (18-28) segment show varying behaviour, but there is a significant contribution of beta-strand-like conformations at all temperatures below 20 degrees C. The C-terminal (29-40) segment was not studied by NMR, but from CD difference spectra we concluded that it is mainly in a random coil conformation at all studied temperatures. These results on structural preferences and transitions of the segments in the monomeric form of A(beta) may be related to the processes leading to the aggregation and formation of fibrils in the Alzheimer plaques. PMID- 16045765 TI - Unravelling the functional interaction structure of a cellular network from temporal slope information of experimental data. AB - Due to the unavoidable nonbiological variations accompanying many experiments, it is imperative to consider a way of unravelling the functional interaction structure of a cellular network (e.g. signalling cascades or gene networks) by using the qualitative information of time-series experimental data instead of computation through the measured absolute values. In this spirit, we propose a very simple but effective method of identifying the functional interaction structure of a cellular network based on temporal ascending or descending slope information from given time-series measurements. From this method, we can gain insight into the acceptable measurement error ranges in order to estimate the correct functional interaction structure and we can also find guidance for a new experimental design to complement the insufficient information of a given experimental dataset. We developed experimental sign equations, making use of the temporal slope sign information from time-series experimental data, without a specific assumption on parameter perturbations for each network node. Based on these equations, we further describe the available specific information from each part of experimental data in detail and show the functional interaction structure obtained by integrating such information. In this procedure, we use only simple algebra on sign changes without complicated computations on the measured absolute values of the experimental data. The result is, however, verified through rigorous mathematical definitions and proofs. The present method provides us with information about the acceptable measurement error ranges for correct estimation of the functional interaction structure and it further leads to a new experimental design to complement the given experimental data by informing us about additional specific sampling points to be chosen for further required information. PMID- 16045766 TI - Local stability identification and the role of a key aromatic amino acid residue in staphylococcal nuclease refolding. AB - Staphylococcal nuclease (SNase) is a model protein that contains one domain and no disulfide bonds. Its stability in the native state may be maintained mainly by key amino acids. In this study, two point-mutated proteins each with a single base substitution [alanine for tryptophan (W140A) and alanine for lysine (K133A)] and two truncated fragment proteins (positions 1-139 [SNase(1-139) or W140O] and positions 1-141 [SNase(1-141) or E142O]) were generated. Differential scanning microcalorimetry in thermal denaturation experiments showed that K133A and E142O have nearly unchanged DeltaH(cal) relative to the wild-type, whereas W140A and W140O display zero enthalpy change (DeltaH(cal) approximately 0). Far-UV CD measurements indicate secondary structure in W140A but not W140O, and near-UV CD measurements indicate no tertiary structure in either W140 mutant. These observations indicate an unusually large contribution of W140 to the stability and structural integrity of SNase. PMID- 16045767 TI - Local stability identification and the role of key acidic amino acid residues in staphylococcal nuclease unfolding. AB - Staphylococcal nuclease is a single domain protein with 149 amino acids. It has no disulfide bonds, which makes it a simple model for the study of protein folding. In this study, 20 mutants of this protein were generated each with a single base substitution of glycine for negatively charged glutamic acid or aspartic acid. Using differential scanning microcalorimetry in thermal denaturation experiments, we identified two mutants, E75G and E129G, having approximately 43% and 44%, respectively, lower DeltaH(cal) values than the wild type protein. Furthermore, two mutants, E75Q and E129Q, were created and the results imply that substitution of the Gly residue has little influence on destabilization of the secondary structure that leads to the large perturbation of the tertiary protein structure stability. Two local stable areas formed by the charge-charge interactions around E75 and E129 with particular positively charged amino acids are thus identified as being significant in maintenance of the three dimensional structure of the protein. PMID- 16045768 TI - Two separate regions essential for nuclear import of the hnRNP D nucleocytoplasmic shuttling sequence. AB - Heterogeneous nuclear ribonucleoprotein (hnRNP) D/AUF1 functions in mRNA genesis in the nucleus and modulates mRNA decay in the cytoplasm. Although it is primarily nuclear, it shuttles between the nucleus and cytoplasm. We studied the nuclear import and export of the last exon-encoding sequence common to all its isoforms by its expression as a green fluorescent protein-fusion protein in HeLa cells and by heterokaryon assay. The C-terminal 19-residue sequence (SGYGKVSRRGGHQNSYKPY) was identified as an hnRNP D nucleocytoplasmic shuttling sequence (DNS). In vitro nuclear transport using permeabilized cells indicated that nuclear import of DNS is mediated by transportin-1 (Trn-1). DNS accumulation in the nucleus was dependent on Trn-1, Ran, and energy in multiple rounds of nuclear transport. Use of DNS with deletions, alanine scanning mutagenesis and point mutations revealed that two separate regions (the N-terminal seven residues and the C-terminal two residues) are crucial for in vivo and in vitro transport as well as for interaction with Trn-1. The N- and C-terminal motifs are conserved in the shuttling sequences of hnRNP A1 and JKTBP. PMID- 16045769 TI - The binding of IMP to ribonuclease A. AB - The binding of inosine 5' phosphate (IMP) to ribonuclease A has been studied by kinetic and X-ray crystallographic experiments at high (1.5 A) resolution. IMP is a competitive inhibitor of the enzyme with respect to C>p and binds to the catalytic cleft by anchoring three IMP molecules in a novel binding mode. The three IMP molecules are connected to each other by hydrogen bond and van der Waals interactions and collectively occupy the B1R1P1B2P0P(-1) region of the ribonucleolytic active site. One of the IMP molecules binds with its nucleobase in the outskirts of the B2 subsite and interacts with Glu111 while its phosphoryl group binds in P1. Another IMP molecule binds by following the retro-binding mode previously observed only for guanosines with its nucleobase at B1 and the phosphoryl group in P(-1). The third IMP molecule binds in a novel mode towards the C-terminus. The RNase A-IMP complex provides structural evidence for the functional components of subsite P(-1) while it further supports the role inferred by other studies to Asn71 as the primary structural determinant for the adenine specificity of the B2 subsite. Comparative structural analysis of the IMP and AMP complexes highlights key aspects of the specificity of the base binding subsites of RNase A and provides a structural explanation for their potencies. The binding of IMP suggests ways to develop more potent inhibitors of the pancreatic RNase superfamily using this nucleotide as the starting point. PMID- 16045770 TI - Insertion of the plant photosystem I subunit G into the thylakoid membrane. AB - Subunit G of photosystem I is a nuclear-encoded protein, predicted to form two transmembrane alpha-helices separated by a loop region. We use in vitro import assays to show that the positively charged loop domain faces the stroma, whilst the N- and C-termini most likely face the lumen. PSI-G constructs in which a His- or Strep-tag is placed at the C-terminus or in the loop region insert with the same topology as wild-type photosystem I subunit G (PSI-G). However, the presence of the tags in the loop make the membrane-inserted protein significantly more sensitive to trypsin, apparently by disrupting the interaction between the loop and the PSI core. Knock-out plants lacking PSI-G were transformed with constructs encoding the C-terminal and loop-tagged PSI-G proteins. Experiments on thylakoids from the transgenic lines show that the C-terminally tagged versions of PSI-G adopt the same topology as wild-type PSI-G, whereas the loop-tagged versions affect the sensitivity of the loop region to trypsin, thus confirming the in vitro observations. Furthermore, purification of PSI complexes from transgenic plants revealed that all the tagged versions of PSI-G are incorporated and retained in the PSI complex, although the C-terminally tagged variants of PSI-G were preferentially retained. This suggests that the loop region of PSI-G is important for proper integration into the PSI core. Our experiments demonstrate that it is possible to produce His- and Strep-tagged PSI in plants, and provide further evidence that the topology of membrane proteins is dictated by the distribution of positive charges, which resist translocation across membranes. PMID- 16045772 TI - Serosal involvement in gastrointestinal cancer: its assessment and significance. AB - There is an increasing burden upon diagnostic histopathologists to identify accurately factors of prognostic and therapeutic implication in gastrointestinal cancer. It is perhaps partly because of the use of rigid sequential staging systems, such as the Dukes' classification, that some factors, perhaps most notably involvement of surgical margins (especially in rectal cancer) and serosal involvement (particularly in oesophageal, colonic and rectal cancer), have been relatively neglected until more recently. This is surprising and concerning because both of these pathologically derived parameters strongly correlate with subsequent locoregional recurrence and, ultimately, with prognosis. Whilst the occurrence and significance of serosal involvement have been well recognized in gastric cancer for many years, relatively little attention has been paid to the phenomenon in oesophageal cancer and yet both pleural and peritoneal involvement may be comparatively commonly identified in oesophageal cancer. Serosal involvement and transperitoneal spread are also of considerable prognostic importance in primary appendiceal carcinoma. Only more recently has the significance of serosal involvement been appreciated in colonic and rectal cancer. In the colon, the phenomenon is now recognized to be one of the most important factors in predicting transperitoneal spread and overall prognosis. Furthermore, there is increasing interest in alternative novel strategies, including intraperitoneal chemotherapy and radical peritoneal surgery, as legitimate therapeutic options in many gastrointestinal cancers. PMID- 16045773 TI - Ki67 expression in different epithelial types in columnar lined oesophagus indicates varying levels of expanded and aberrant proliferative patterns. AB - AIMS: To evaluate proliferative patterns in metaplastic columnar epithelia of the oesophagus, classified as oxynto-cardiac (n = 43), cardiac (n = 45) intestinal without dysplasia (n = 41), dysplastic intestinal epithelium (n = 25), and adenocarcinoma (n = 15) by Ki67 immunohistochemistry. METHODS AND RESULTS: Abnormal patterns of Ki67 immunoreactivity were classified into (i) expanded proliferation, characterized by increased levels of Ki67 expression in the deep and mid third of the foveolar pit; and (ii) aberrant proliferation, characterized by positive staining in the surface epithelium and superficial third of the foveolar pit. A significant step-wise increase in the frequency of expanded proliferation was seen in oxynto-cardiac, cardiac, intestinal and dysplastic intestinal epithelium indicative of increasing levels of damage. Aberrant proliferation was absent in oxynto-cardiac mucosa, present at a low and similar level in cardiac, intestinal and low-grade dysplastic epithelia and at a significantly increased frequency in high-grade dysplasia. CONCLUSIONS: These findings suggest that oxynto-cardiac mucosa occurs in a low damage environment and intestinal metaplasia in a high damage environment along the length of the columnar lined oesophageal segment. Aberrant proliferative patterns with Ki67 staining are not useful in differentiating reactive epithelia from low-grade dysplasia, but may prove useful in the diagnosis of high-grade dysplasia. PMID- 16045774 TI - Pathological response following long-course neoadjuvant chemoradiotherapy for locally advanced rectal cancer. AB - AIMS: To standardize the pathological analysis of total mesorectal excision specimens of rectal cancer following neoadjuvant chemoradiotherapy for locally advanced disease (T3/T4), including tumour regression. METHODS AND RESULTS: Standardized dissection and reporting was used for 60 patients who underwent total mesorectal excision following long-course chemoradiotherapy. Tumour regression was scored by two pathologists (K.S., D.G.) using both an established 5-point tumour regression grade (TRG), and a novel 3-point grade. Both scores were evaluated for interobserver variability. A complete or near-complete pathological response (3-point TRG 1) was found in 10 patients (17%). Using the 5 point TRG, there was good agreement between both pathologists (kappa = 0.64). Using the 3-point grade, agreement was excellent (kappa = 0.84). No disease recurrence has been reported in patients with a complete, or near complete pathological response (3-point TRG 1), after a mean follow-up of 22 months. CONCLUSION: Tumour regression grade is a useful method of scoring tumour response to chemoradiotherapy in rectal cancer. TRG 1 and 2 can be regarded as a complete pathological response (ypT0). A modified 3-point grade has the advantage of better reproducibility, with similar prognostic significance. PMID- 16045775 TI - Immunohistochemical study of autoimmune pancreatitis using anti-IgG4 antibody and patients' sera. AB - AIMS: Autoimmune pancreatitis (AIP), characterized by raised serum IgG4 levels, is frequently complicated by disorders of extrapancreatic organs. The aim of the present study was to examine immunohistochemically which extrapancreatic organs are affected, and whether an autoantibody to such organs is present in the serum of AIP patients. METHODS: Various tissues/organs obtained from AIP patients were studied immunohistochemically with an anti-IgG4 antibody. To examine the presence of an autoantibody in the serum of AIP patients, sera were incubated with various normal organs/tissues extracted for other diseases, followed by detection with an anti-IgG4 antibody. Sera were also examined before and after glucocorticoid therapy. RESULTS: Marked infiltration of IgG4+ plasma cells was observed in the pancreas, liver, bile duct and salivary gland of many of the AIP patients examined. The normal epithelia of the pancreatic ducts, bile ducts, gallbladder and salivary gland ducts reacting with the patients' sera were detectable by the anti-IgG4 antibody. Following glucocorticoid therapy the IgG4 antibody from the patients' sera showed decreased reactivity with these tissues. CONCLUSIONS: AIP may also affect extrapancreatic organs, the serum of AIP patients may contain an IgG4 autoantibody to various organs and glucocorticoid therapy may improve such disorders. PMID- 16045776 TI - Usefulness of epithelial membrane antigen (EMA) to discriminate between perineural invasion and perineural indentation in prostatic carcinoma. AB - AIMS: Perineural invasion (PNI) is one of the few unequivocal criteria to diagnose adenocarcinoma of the prostate. Distinguishing PNI from perineural indentation (PNIn), however, is sometimes difficult. The aim of this study was to discriminate between PNI and PNIn using EMA immunohistochemistry. METHODS AND RESULTS: We selected representative sections from 87 prostatectomies with prostatic adenocarcinoma. Normal peripheral nerves were continuously encircled with perineurium, which was immunoreactive for EMA. We identified 1319 PNI by carcinomas, 368 PNIn by carcinomas, and 303 PNIn by benign glands. We categorized the EMA immunoreactivity patterns into three classes: samples that displayed discontinuity or complete loss of the perineurium (Type A), samples where there were carcinomas or benign glands in the perineural space or peripheral nerves (Type B) and samples that showed no changes in the perineurium (Type C). For PNI we observed Type A, Type B, and Type C patterns in 55.3%, 24.8% and 19.9% of carcinomas, respectively. The incidence of each of those patterns in PNIn by carcinomas was 32.1%, 14.9% and 53.0%, respectively. Cases of PNIn by benign glands showed Type A or Type C patterns. They did not, however, exhibit Type B patterns. CONCLUSION: EMA immunostaining will aid the diagnosis of prostatic adenocarcinoma. PMID- 16045777 TI - The continued value of central histopathological review of testicular tumours. AB - AIMS: Central histopathological review of testicular tumours prior to definitive treatment can have an important impact on patient management. This study was designed to assess the continued value of central review in the light of increasing subspecialization and increased numbers of consultant histopathologists. MATERIALS AND RESULTS: The original and review reports of 291 testicular cancer specimens from 1998 to 2002 were analysed, looking particularly at major diagnosis, vascular invasion and the tumour elements within non seminomatous germ cell tumours (NSGCT). When a diagnosis was altered any effect on subsequent patient management was assessed. There was a discrepancy in tumour type in 11 cases (4%) compared with 6% in 1992-1997. The commonest change was from seminoma to NSGCT or combined germ cell tumour (5/11). There was also diagnostic difficulty with spermatocytic seminoma (3/11). The clinical management of all 11 cases was influenced as a result of the review diagnosis. Discrepancies in vascular invasion were noted in 13 of the 126 NSGCTs (10%) compared with 20% in 1992-1997. Differences in NSGCT tumour elements, though clinically less important, were frequent in both groups. CONCLUSIONS: There continues to be a small number of significant and clinically important errors identified following central histopathological review of testicular tumours. This study highlights the value of central review and supports its continued practice in the management of testicular tumours. PMID- 16045778 TI - Differentiating keratoacanthoma from squamous cell carcinoma by the use of apoptotic and cell adhesion markers. AB - AIMS: Keratoacanthomas (KA) are well-differentiated squamoproliferative skin lesions that grow rapidly and regress spontaneously. In contrast, squamous cell carcinomas (SCC) can have variable differentiation, inexorably progress and on occasion metastasize. Distinguishing between KA and SCC using haematoxylin and eosin-stained sections from an initial biopsy can often be difficult. There is also some debate as to whether KA is simply a variety of well-differentiated SCC or a distinct entity. METHODS AND RESULTS: Initial biopsy sections from 25 cases of SCC and 20 of KA were labelled with markers for both the initiation (the cytolytic receptor P2X7) and end-stage (terminal deoxynucleotidyl transferase biotin-dUTP nick end labelling) of apoptosis, telomerase-associated protein (TP1) and the cell adhesion protein E-cadherin. As this was a retrospective study, the clinical outcome of each case was known. This resulted in a unique labelling pattern of each marker for SCC and KA, allowing a differential diagnosis between the two conditions. The simplest marker to use for this purpose was anti-P2X7. Sections from five cases that were initially very difficult to diagnose were correctly identified as SCC using this method. CONCLUSIONS: These results support the view that KA has a different pathogenesis and biochemistry from that of SCC, and is a distinct entity. Anti-P2X7 labelling, using routine immunohistochemical techniques, provides a method for differentially diagnosing these conditions. PMID- 16045779 TI - Dermal non-neural granular cell tumour (so-called primitive polypoid granular cell tumour): a distinctive entity further delineated in a clinicopathological study of 11 cases. AB - AIMS: Cutaneous and soft tissue granular cell tumour is a well-characterized benign neoplasm of neural origin. However, there remains a subcategory of granular cell tumour, first described by Le Boit as 'primitive polypoid granular cell tumour', that shows no obvious line of differentiation. The aim of this study is to further the characterization of this lesion by undertaking a clinicopathological review. METHODS AND RESULTS: Eleven cases of dermal non neural granular cell tumour were retrieved from one of the authors referral archives (E.C.) and both the histology and immunohistochemistry reviewed. Clinical data with follow-up were obtained from the referring pathologists. The lesions most commonly occurred in young to middle-aged adults (nine cases, median = 33 years, age range 6-56 years), with a slight female predominance. They presented as painless nodules, mainly on the extremities or face. Local excision was the treatment of choice and up to date follow-up reveals no sign of recurrence. Histologically, eight cases were polypoid, while three cases were endophytic. The tumours were composed of elongated spindle-shaped to polygonal or round cells with prominent granular cell change, and tumour nuclei showing mild focal atypia to rare moderate atypia. Mitotic activity ranged from one to nine mitoses per 10 high-power fields (median = 2, mean = 3.8). Immunohistochemical labelling of the tumour cells demonstrated expression for NKI-C3 (n = 11), focal, weak positivity for CD68 (n = 10) and FXIIIa (n = 2). There was negative staining for S100 protein, smooth muscle actin, Melan-A, CD34, desmin and cytokeratin. CONCLUSIONS: This analysis of 11 cases contributes to the characterization of this recently described entity, which despite some atypical histological features and no obvious line of differentiation, behaves in a completely indolent fashion. PMID- 16045780 TI - Microvessel density for melanoma prognosis. AB - AIMS: Two-hundred and four accessible cases of malignant melanoma from the Grampian region of Scotland, collected over a period of 4 years, were studied for standard prognostic indicators for comparison with microvessel density. METHODS AND RESULTS: The range of tumour thickness varied from in-situ melanoma to 14.8 mm. Microvessel density was assessed using the Chalkley technique on sections immunostained with CD31 antibody to identify endothelium. Vessel counts were performed in the peritumoral host tissue of all specimens. Strong correlation was observed between microvessel density at the tumour edge and tumour thickness (P < 0.001). Multifactorial regression analysis confirms Chalkley estimation as a reliable predictor of the risk of recurrence of melanoma (P < 0.005). The predictive value was found to be superior to the Breslow score, for tumours more than 2 mm thick. CONCLUSIONS: Microvessel assessment of primary melanoma using the Chalkley score technique provides reliable prognostic information on the risk of recurrence of the tumour, particularly for melanomas deeper than 2 mm. It remains to be seen whether this methodology can predict response to novel anti angiogenic therapies currently entering trials. PMID- 16045781 TI - Expression of GCDFP-15 and AR decreases in larger or node-positive apocrine carcinomas of the breast. AB - AIMS: Apocrine carcinoma of the breast is typically, though not always, positive for gross cystic disease fluid protein-15 (GCDFP-15). In order to clarify the clinical significance of GCDFP-15 in apocrine carcinomas, GCDFP-15 expression was examined in apocrine carcinomas of different stages and compared with clinicopathological factors. Apocrine lesions reportedly exhibit an unusual immunohistochemical status, expressing androgen receptors (AR) instead of oestrogen receptors (ER), progesterone receptors (PR), or bcl-2. Their expression was also examined. METHODS AND RESULTS: Fifty-two apocrine carcinomas were examined immunohistochemically. Thirty-nine (75%) and 29 (56%) were positive for GCDFP-15 and AR, respectively. GCDFP-15 positivity was significantly lower in infiltrating carcinomas than intraductal carcinomas (P = 0.0111). In infiltrating carcinomas, GCDFP-15 positivity was significantly low in tumours > or = 15 mm (P = 0.0005) and node-positive tumours (P = 0.0004). Similar phenomena were observed for AR. Rare cases were positive for ER (3.8%), PR (5.8%), and bcl-2 (1.9%). CONCLUSIONS: GCDFP-15 positivity is transient and should not be considered a definitive marker of apocrine carcinomas. Cases which have apocrine features but lack GCDFP-15 expression should rather be considered as advanced apocrine carcinomas. ER/PR/bcl-2 negativity will sometimes be helpful to confirm the diagnosis of apocrine carcinoma, because it is more consistent than GCDFP-15/AR positivity. PMID- 16045782 TI - Observer variability in the histopathological reporting of core biopsies of papillary breast lesions is reduced by the use of immunohistochemistry for CK5/6, calponin and p63. AB - AIM: To investigate agreement on core biopsy diagnosis of papillary breast lesions, which is acknowledged as a difficult area, and to determine the effect of the use of immunohistochemistry (IHC) to assist diagnosis. STUDY DESIGN: Haematoxylin and eosin (H&E) sections of 129 core biopsies of papillary breast lesions were circulated to four observers who categorized each case as: B2 (benign), B3a (epithelial proliferation, probably benign but requiring biopsy), B3b (epithelial proliferation with cytological or architectural atypia), B4 (probably malignant but insufficient material or artefact to allow diagnosis), B5 (malignant papillary lesion). In all cases (n = 127) IHC was performed for cytokeratin (CK) 5/6, calponin, p63 (myoepithelial markers), and slides recirculated. RESULTS: There was unanimous agreement in 44% of cases on H&E only which rose to 91% after the use of IHC. Overall, unweighted kappa (Ku; five categories) rose from 0.54 to 0.91. The main effect of IHC was to reduce the use of intermediate categories (B3a, B3b and B4) and allow definitive diagnosis (B2 or B5). CONCLUSION: Agreement on H&E sections alone in papillary core biopsies of breast is only 44% (Ku = 0.54) but is significantly increased to 91% (Ku = 0.91) by the use of IHC for CK5/6, calponin and p63. PMID- 16045783 TI - Galectin-3 expression in papillary microcarcinoma of the thyroid. AB - AIMS: Galectin-3 is a beta-galactoside binding protein, recently recognized as a promising molecular marker of thyroid malignancy. As reported in several studies, galectin-3 is highly expressed in papillary thyroid carcinoma, but its expression has not been investigated in papillary microcarcinoma, which is a variant of papillary thyroid carcinoma. METHODS AND RESULTS: Using a monoclonal antibody to galectin-3 and the avidin-biotin-peroxidase complex (ABC) immunohistochemical technique, we analysed galectin-3 expression in 63 cases of papillary microcarcinoma. The results showed immunohistochemical reactivity for galectin-3 in 51 (80.9%) cases. Intensity of staining varied from strong or moderate to weak. Galectin-3 localization was mostly cytoplasmic, but also membranous or nuclear in some cells. Immunohistochemical expression of galectin-3 was not found in 12 (19.1%) cases. Most galectin-3 negative microcarcinomas (10/12) were of the non-classical type, i.e. without papillary architecture. Neither the frequency nor the intensity of a positive reaction was related to tumour size. CONCLUSIONS: Galectin-3 gene is expressed at the protein level in most papillary microcarcinomas, although with slightly lower frequency than that reported for clinically evident papillary thyroid carcinoma. The presence of galectin-3 in clinically silent microcarcinomas may indicate that galectin-3 is not related to growth or aggressiveness of papillary thyroid microcarcinomas but rather plays some other role in thyroid tumour biology. PMID- 16045784 TI - Spontaneous coronary artery dissection. PMID- 16045785 TI - Metastatic extramammary Paget's disease with HER2 amplification to renal cell carcinoma: a unique presentation of cancer in cancer metastasis. PMID- 16045786 TI - A novel entity: low-grade mucinous tubular renal carcinoma. PMID- 16045787 TI - 'Secretory endometrioid-like' adenocarcinoma of the lung. PMID- 16045788 TI - Nodal peripheral T-cell lymphoma associated with Warthin's tumour. PMID- 16045789 TI - Mucormycosis of the stomach in a patient with multiple trauma. PMID- 16045790 TI - Rhinoscleroma of the larynx. PMID- 16045791 TI - Number of levels needed for diagnosis of endometrial biopsies. PMID- 16045792 TI - Effect of short schemes on body composition measurements using air-displacement plethysmography. AB - BACKGROUND: Air-displacement plethysmography (ADP) is becoming a popular method to assess body composition. Several studies have shown certain types of clothing can affect measurements of body density, however no study has specifically investigated the effect of cotton gym shorts and spandex bicycle shorts on body density. METHODS: Thirty-seven males (23.0 +/- 3.2 yr., 177.3 +/- 5.4 cm., 74.8 +/- 7.5 kg.) and thirty-eight females (23.7 +/- 5.3 yr., 163.6 +/- 8.4 cm., 57.1 +/- 7.0 kg.) had their body density measured by ADP in three clothing schemes: 1) a tight fitting Speedo swim suit (criterion measure), 2) cotton gym shorts, and 3) spandex bicycle shorts. The clothing was provided by the University of Oklahoma Body Composition Laboratory and the testing schemes were performed in random order. RESULTS: The regression of body density by the criterion measure against body density while wearing cotton gym shorts for the entire group (y = 0.001 + 0.991x, SEE = 0.003 g/cm3) and the females (y = 0.059 + 0.934x, SEE = 0.003 g/cm3) did not significantly deviate from the line of identity. However in males the regression significantly deviated from the line of identity (y = 0.052 + 0.944x, SEE = 0.002 g/cm3). Body density by the criterion measure and body density while wearing spandex bicycle shorts did not significantly differ from the line of identity for the entire group (y = -0.018 + 1.013x SEE = 0.003 g/cm3), in males (y = -0.002 + 1.001x, SEE = 0.003 g/cm3), or females (y = 0.073 + 0.925x, SEE = 0.003 g/cm3). Residual plot analysis revealed no group or gender bias in either the cotton gym shorts or in the spandex bicycle shorts. CONCLUSION: It would appear bicycle spandex shorts are an acceptable alternative to a Speedo like swim suit, however we advise that subjects adhere to the strict clothing protocol that is recommended by the manufacturer. PMID- 16045793 TI - A novel combination of multiple primary carcinomas: urinary bladder transitional cell carcinoma, prostate adenocarcinoma and small cell lung carcinoma--report of a case and review of the literature. AB - BACKGROUND: The incidence of multiple primary malignant neoplasms increases with age and they are encountered more frequently nowadays than before, the phenomenon is still considered to be rare. CASE PRESENTATION: We report a case of a man in whom urinary bladder transitional cell carcinoma, metachronous prostate adenocarcinoma and small cell lung carcinoma were diagnosed within an eighteen month period. The only known predisposing factor was that he was heavy smoker (90 100 packets per year). The literature on the phenomenon of multiple primary malignancies in a single patient is reviewed and the data is summarized. CONCLUSION: It is important for the clinicians to keep in mind the possibility of a metachronous (successive) or a synchronous (simultaneous) malignancy in a cancer patient. It is worthy mentioning this case because clustering of three primary malignancies (synchronous and metachronous) is of rare occurrence in a single patient, and, to our knowledge, this is the first report this combination of three carcinomas appearing in the same patient. PMID- 16045794 TI - Genotyping of Mycobacterium tuberculosis clinical isolates in two cities of Turkey: description of a new family of genotypes that is phylogeographically specific for Asia Minor. AB - BACKGROUND: Population-based bacterial genetics using repeated DNA loci is an efficient approach to study the biodiversity and phylogeographical structure of human pathogens, such as Mycobacterium tuberculosis, the agent of tuberculosis. Indeed large genetic diversity databases are available for this pathogen and are regularly updated. No population-based polymorphism data were yet available for M. tuberculosis in Turkey, at the crossroads of Eurasia. RESULTS: A total of 245 DNAs from Mycobacterium tuberculosis clinical isolates from tuberculosis patients residing in Turkey (Malatya n = 147 or Ankara n = 98) were genotyped by spoligotyping, a high-throughput genotyping method based on the polymorphism of the Direct Repeat locus. Thirty-three spoligotyping-defined clusters including 206 patients and 39 unique patterns were found. The ST41 cluster, as designated according to the international SpolDB3 database project, represented one fourth and when gathered to three genotypes, ST53, ST50 and ST284, one half of all the isolates. Out of 34 clinical isolates harboring ST41 which were further genotyped by IS6110 and by MIRU-VNTR typing, a typical 2-copy IS6110-RFLP pattern and a "215125113322" MIRU-VNTR pattern were observed among 21 clinical isolates. Further search in various databases confirms the likely Turkish-phylogeographical specificity of this clonal complex. CONCLUSION: We described a new phylogeographically-specific clone of M. tuberculosis, designated LAM7-TUR. Further investigations to assess its frequency within all regions of Turkey and its phylogeographical origin and phylogenetic position within the global M. tuberculosis phylogenetic tree will shed new light on its endemicity in Asia Minor. PMID- 16045795 TI - The epidemiology of low back pain in primary care. AB - This descriptive review provides a summary of the prevalence, activity limitation (disability), care-seeking, natural history and clinical course, treatment outcome, and costs of low back pain (LBP) in primary care. LBP is a common problem affecting both genders and most ages, for which about one in four adults seeks care in a six-month period. It results in considerable direct and indirect costs, and these costs are financial, workforce and social. Care-seeking behaviour varies depending on cultural factors, the intensity of the pain, the extent of activity limitation and the presence of co-morbidity. Care-seeking for LBP is a significant proportion of caseload for some primary-contact disciplines. Most recent-onset LBP episodes settle but only about one in three resolves completely over a 12-month period. About three in five will recur in an on-going relapsing pattern and about one in 10 do not resolve at all. The cases that do not resolve at all form a persistent LBP group that consume the bulk of LBP compensable care resources and for whom positive outcomes are possible but not frequent or substantial. PMID- 16045796 TI - Is neutrophil elastase the missing link between emphysema and fibrosis? Evidence from two mouse models. AB - BACKGROUND: The separation of emphysema from fibrosis is not as clear-cut as it was thought in early studies. These two pathologies may be present at the same time in human lungs and in mice either instilled with elastolytic enzymes or bleomycin or exposed to cigarette-smoke. According to a current view, emphysema originates from a protease/antiprotease imbalance, and a role for antiproteases has also been suggested in the modulation of the fibrotic process. In this study we investigate in experimental animal models of emphysema and fibrosis whether neutrophil elastase may constitute a pathogenic link between these two pathologies. METHODS: This study was done in two animal models in which emphysema and fibrosis were induced either by bleomycin (BLM) or by chronic exposure to cigarette-smoke. In order to assess the protease-dependence of the BLM-induced lesion, a group mice was treated with 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride, a serine proteinase inhibitor active toward neutrophil elastase. Lungs from each experimental group were used for the immunohistochemical assessment of transforming growth factor-beta (TGF-beta) and transforming growth factor-alpha (TGF-alpha) and for determination of the mean linear intercept as well as the percent volume densities of fibrosis and of emphysematous changes. Additionally, the lungs were also assessed for desmosine content and for the determination of elastase levels in the pulmonary interstitium by means of immunoelectron microscopy. RESULTS: We demonstrate that in BLM-treated mice (i) the development of elastolytic emphysema precedes that of fibrosis; (ii) significant amount of elastase in alveolar interstitium is associated with an increased expression of TGF-beta and TGF-alpha; and finally, (iii) emphysematous and fibrotic lesions can be significantly attenuated by using a protease inhibitor active against neutrophil elastase. Also, in a strain of mice that develop both emphysema and fibrosis after chronic cigarette-smoke exposure, the presence of elastase in alveolar structures is associated with a positive immunohistochemical reaction for reaction for both TGF-beta and TGF-alpha. CONCLUSION: The results of the present study strongly suggest that neutrophil elastase may represent a common pathogenic link between emphysema and fibrosis. Proteases and in particular neutrophil elastase could act as regulatory factors in the generation of soluble cytokines with mitogenic activity for mesenchymal cells resulting either in emphysema or in fibrosis or both. PMID- 16045797 TI - The color of a Dalmatian's spots: linkage evidence to support the TYRP1 gene. AB - BACKGROUND: The distinctive coat pattern of a Dalmatian is the result of the interaction of several loci. While the encoded function of these genes is not fully understood, it is known the Piebald, Ticking, and Flecking loci interact to produce the Dalmatian's classic pigmented spots on a white background. The color of the pigmented spots in purebred Dalmatians can either be black or liver, but the locus responsible for color determination is unknown. Studies have been conducted to determine the underlying genes involved in coat color determination in the dog, e.g., in the Labrador Retriever, but none to date have addressed black versus liver in the Dalmatian. RESULTS: A genome scan was conducted in a multi-generational kindred of Dalmatians segregating black and liver spot color. Linkage analysis was performed using a total of 113 polymorphic microsatellite markers from the kindred. Linkage was found between spot color and a single microsatellite marker, FH2319 (LOD = 12.5) on chromosome 11. CONCLUSION: The TYRP1 (Brown) locus is located at position 50.1 Mb on chromosome 11, which is approximately 0.4 Mb from marker FH2319. Given the recent characterization of TYRP1 genetic variations in the dog and the linkage evidence reported here, TYRP1 is likely responsible for the spot color variation of black versus liver seen in the Dalmatian. PMID- 16045798 TI - Induction of compulsive-like washing by blocking the feeling of knowing: an experimental test of the security-motivation hypothesis of obsessive-compulsive disorder. AB - BACKGROUND: H. Szechtman and E. Woody (2004) hypothesized that obsessive compulsive disorder results from a deficit in the feeling of knowing that normally terminates thoughts or actions elicited by security motivation. To test the plausibility of this proposed mechanism, an experiment was conducted to produce an analog of washing in obsessive-compulsive disorder by eliciting a scenario of potential harm and using hypnosis to block changes in internally generated feelings that would normally occur during washing. RESULTS: Participants reacted with increased disgust, anxiety, and heart rate to their mental images of contamination and potential danger. As predicted, high but not low hypnotizable participants showed a significant prolongation of washing when change in feelings during washing was blocked hypnotically. CONCLUSION: Results show that blocking the affective signal that is normally generated during security-related behaviors, such as washing, leads to prolonged performance of these behaviors. This finding lends support to the plausibility of the proposed model of obsessive-compulsive disorder. PMID- 16045799 TI - A generic RNA-pulsed dendritic cell vaccine strategy for renal cell carcinoma. AB - We present a generic dendritic cell (DC) vaccine strategy for patients with renal cell carcinoma (RCC) based on the use of RNA as a source of multiplex tumor associated antigens (TAAs). Instead of preparing RNA from tumor tissue of each individual RCC patient, we propose to substitute RNA prepared from a well characterized highly immunogenic RCC cell line (RCC-26 tumor cells) as a generic source of TAAs for loading of DCs. We demonstrate here that efficient RNA transfer can be achieved using lipofection of immature DCs, which are subsequently matured with a cytokine cocktail to express high levels of MHC and costimulatory molecules as well as the chemokine receptor CCR7. Neither RNA itself nor the lipid component impacted on the phenotype or the cytokine secretion of mature DCs. Following RNA loading, DCs derived from HLA-A2-positive donors were able to activate effector-memory cytotoxic T lymphocytes (CTLs) specific for a TAA ligand expressed by the RCC-26 cell line. CTL responses to RNA loaded DCs reached levels comparable to those stimulated directly by the RCC-26 tumor cells. Furthermore, DCs expressing tumor cell RNA primed naive T cells, yielding T cell lines with cytotoxicity and cytokine secretion after contact with RCC tumor cells. RCC-26 cell lines are available as good manufacturing practice (GMP)-certified reagents enabling this source of RNA to be easily standardized and adapted for clinical testing. In addition, well defined immune monitoring tools, including the use of RNA expressing B cell lines, are available. Thus, this DC vaccine strategy can be directly compared with an ongoing gene therapy trial using genetically-engineered variants of the RCC-26 cell line as vaccines for RCC patients with metastatic disease. PMID- 16045801 TI - Can context justify an ethical double standard for clinical research in developing countries? AB - BACKGROUND: The design of clinical research deserves special caution so as to safeguard the rights of participating individuals. While the international community has agreed on ethical standards for the design of research, these frameworks still remain open to interpretation, revision and debate. Recently a breach in the consensus of how to apply these ethical standards to research in developing countries has occurred, notably beginning with the 1994 placebo controlled trials to reduce maternal to child transmission of HIV-1 in Africa, Asia and the Caribbean. The design of these trials sparked intense debate with the inclusion of a placebo-control group despite the existence of a 'gold standard' and trial supporters grounded their justifications of the trial design on the context of scarcity in resource-poor settings. DISCUSSION: These 'contextual' apologetics are arguably an ethical loophole inherent in current bioethical methodology. However, this convenient appropriation of 'contextual' analysis simply fails to acknowledge the underpinnings of feminist ethical analysis upon which it must stand. A more rigorous analysis of the political, social, and economic structures pertaining to the global context of developing countries reveals that the bioethical principles of beneficence and justice fail to be met in this trial design. CONCLUSION: Within this broader, and theoretically necessary, understanding of context, it becomes impossible to justify an ethical double standard for research in developing countries. PMID- 16045800 TI - Mesenchymal stem cells avoid allogeneic rejection. AB - Adult bone marrow derived mesenchymal stem cells offer the potential to open a new frontier in medicine. Regenerative medicine aims to replace effete cells in a broad range of conditions associated with damaged cartilage, bone, muscle, tendon and ligament. However the normal process of immune rejection of mismatched allogeneic tissue would appear to prevent the realisation of such ambitions. In fact mesenchymal stem cells avoid allogeneic rejection in humans and in animal models. These finding are supported by in vitro co-culture studies. Three broad mechanisms contribute to this effect. Firstly, mesenchymal stem cells are hypoimmunogenic, often lacking MHC-II and costimulatory molecule expression. Secondly, these stem cells prevent T cell responses indirectly through modulation of dendritic cells and directly by disrupting NK as well as CD8+ and CD4+ T cell function. Thirdly, mesenchymal stem cells induce a suppressive local microenvironment through the production of prostaglandins and interleukin-10 as well as by the expression of indoleamine 2,3,-dioxygenase, which depletes the local milieu of tryptophan. Comparison is made to maternal tolerance of the fetal allograft, and contrasted with the immune evasion mechanisms of tumor cells. Mesenchymal stem cells are a highly regulated self-renewing population of cells with potent mechanisms to avoid allogeneic rejection. PMID- 16045802 TI - Therapeutic Electromagnetic Field (TEMF) and gamma irradiation on human breast cancer xenograft growth, angiogenesis and metastasis. AB - BACKGROUND: The effects of a rectified semi-sinewave signal (15 mT amplitude, 120 pulses per second, EMF Therapeutics, Inc.) (TEMF) alone and in combination with gamma irradiation (IR) therapy in nude mice bearing a human MDA MB231 breast cancer xenograft were tested. Green fluorescence protein transfected cancer cells were injected into the mammary fat pad of young female mice. Six weeks later, mice were randomly divided into four treatment groups: untreated controls; 10 minute daily TEMF; 200 cGy of IR every other day (total 800 cGy); IR plus daily TEMF. Some mice in each group were euthanized 24 hours after the end of IR. TEMF treatment continued for 3 additional weeks. Tumor sections were stained for: endothelial cells with CD31 and PAS or hypoxia inducible factor 1alpha (HIF). RESULTS: Most tumors <35 mm3 were white but tumors >35 mm3 were pink and had a vascularized capsule. The cortex within 100 microns of the capsule had little vascularization. Blood vessels, capillaries, and endothelial pseudopods were found at >100 microns from the capsule (subcortex). Tumors >35 mm3 treated with IR 24 hours previously or with TEMF had decreased blood vessels in the subcortex and more endothelial pseudopods projecting into hypoxic, HIF positive areas than tumors from the control group. Mice that received either IR or TEMF had significantly fewer lung metastatic sites and slower tumor growth than did untreated mice. No harmful side effects were attributed to TEMF. CONCLUSION: TEMF therapy provided a safe means for retarding tumor vascularization, growth and metastasis. PMID- 16045803 TI - Evaluation of normalization methods for cDNA microarray data by k-NN classification. AB - BACKGROUND: Non-biological factors give rise to unwanted variations in cDNA microarray data. There are many normalization methods designed to remove such variations. However, to date there have been few published systematic evaluations of these techniques for removing variations arising from dye biases in the context of downstream, higher-order analytical tasks such as classification. RESULTS: Ten location normalization methods that adjust spatial- and/or intensity dependent dye biases, and three scale methods that adjust scale differences were applied, individually and in combination, to five distinct, published, cancer biology-related cDNA microarray data sets. Leave-one-out cross-validation (LOOCV) classification error was employed as the quantitative end-point for assessing the effectiveness of a normalization method. In particular, a known classifier, k nearest neighbor (k-NN), was estimated from data normalized using a given technique, and the LOOCV error rate of the ensuing model was computed. We found that k-NN classifiers are sensitive to dye biases in the data. Using NONRM and GMEDIAN as baseline methods, our results show that single-bias-removal techniques which remove either spatial-dependent dye bias (referred later as spatial effect) or intensity-dependent dye bias (referred later as intensity effect) moderately reduce LOOCV classification errors; whereas double-bias-removal techniques which remove both spatial- and intensity effect reduce LOOCV classification errors even further. Of the 41 different strategies examined, three two-step processes, IGLOESS-SLFILTERW7, ISTSPLINE-SLLOESS and IGLOESS-SLLOESS, all of which removed intensity effect globally and spatial effect locally, appear to reduce LOOCV classification errors most consistently and effectively across all data sets. We also found that the investigated scale normalization methods do not reduce LOOCV classification error. CONCLUSION: Using LOOCV error of k-NNs as the evaluation criterion, three double-bias-removal normalization strategies, IGLOESS SLFILTERW7, ISTSPLINE-SLLOESS and IGLOESS-SLLOESS, outperform other strategies for removing spatial effect, intensity effect and scale differences from cDNA microarray data. The apparent sensitivity of k-NN LOOCV classification error to dye biases suggests that this criterion provides an informative measure for evaluating normalization methods. All the computational tools used in this study were implemented using the R language for statistical computing and graphics. PMID- 16045804 TI - Resistance to aspirin is increased by ST-elevation myocardial infarction and correlates with adenosine diphosphate levels. AB - BACKGROUND: To be fully activated platelets are dependent on two positive feedback loops; the formation of thromboxane A2 by cyclooxygenase in the platelets and the release of ADP. We wanted to evaluate the effect of aspirin on platelet function in patients with acute coronary syndromes and we hypothesized that increased levels of ADP in patients with acute coronary syndromes could contribute to aspirin resistance. METHODS: Platelet activity in 135 patients admitted for chest pain was assessed with PFA-100. An epinephrine-collagen cartridge (EPI-COLL) was used for the detection of aspirin resistance together with an ADP-collagen cartridge (ADP-COLL). ADP was measured with hplc from antecubital vein samples. Three subgroups were compared: chest pain with no sign of cardiac disease (NCD), NonST-elevation myocardial infarction (NSTEMI) and STEMI. RESULTS: Platelet activation was increased for the STEMI group compared NCD. Aspirin resistance defined as <193 sec in EPI-COLL was 9.7 % in NCD, and increased to 26.0 % (n.s.) in NSTEMI and 83.3 % (p < 0.001) in STEMI. Chronic aspirin treatment significantly reduced platelet aggregation in NCD and NSTEMI, but it had no effect in STEMI. Plasma levels of ADP were markedly increased in STEMI (905 +/- 721 nmol/l, p < 0.01), but not in NSTEMI (317 +/- 245), compared to NCD (334 +/- 271, mean +/- SD). ADP levels correlated with increased platelet activity measured with ADP-COLL (r = -0.30, p < 0.05). Aspirin resistant patients (EPI-COLL < 193 sec) had higher ADP levels compared to aspirin responders (734 +/ 807 vs. 282 +/- 187 nmol/l, mean +/- SD, p < 0.05). CONCLUSION: Platelets are activated and aspirin resistance is more frequent in STEMI, probably due to a general activation of platelets. ADP levels are increased in STEMI and correlates with platelet activation. Increased levels of ADP could be one reason for increased platelet activity and aspirin resistance. PMID- 16045805 TI - Internet-based search of randomised trials relevant to mental health originating in the Arab world. AB - BACKGROUND: The internet is becoming a widely used source of accessing medical research through various on-line databases. This instant access to information is of benefit to busy clinicians and service users around the world. The population of the Arab World is comparable to that of the United States, yet it is widely believed to have a greatly contrasting output of randomised controlled trials related to mental health. This study was designed to investigate the existence of such research in the Arab World and also to investigate the availability of this research on-line. METHODS: Survey of findings from three internet-based potential sources of randomised trials originating from the Arab world and relevant to mental health care. RESULTS: A manual search of an Arabic online current contents service identified 3 studies, MEDLINE, EMBASE, and PsycINFO searches identified only 1 study, and a manual search of a specifically indexed, study-based mental health database, PsiTri, revealed 27 trials. CONCLUSION: There genuinely seem to be few trials from the Arab world and accessing these on-line was problematic. Replication of some studies that guide psychiatric/psychological practice in the Arab world would seem prudent. PMID- 16045806 TI - In normal rat, intraventricularly administered insulin-like growth factor-1 is rapidly cleared from CSF with limited distribution into brain. AB - BACKGROUND: Putatively active drugs are often intraventricularly administered to gain direct access to brain and circumvent the blood-brain barrier. A few studies on the normal central nervous system (CNS) have shown, however, that the distribution of materials after intraventricular injections is much more limited than presumed and their exit from cerebrospinal fluid (CSF) is more rapid than generally believed. In this study, we report the intracranial distribution and the clearance from CSF and adjacent CNS tissue of radiolabeled insulin-like growth factor-1 after injection into one lateral ventricle of the normal rat brain. METHODS: Under barbiturate anesthesia, 125I-labeled insulin-like growth factor-1 (IGF-1) was injected into one lateral ventricle of normal Sprague-Dawley rats. The subsequent distribution of IGF-1 through the cerebrospinal fluid (CSF) system and into brain, cerebral blood vessels, and systemic blood was measured over time by gamma counting and quantitative autoradiography (QAR). RESULTS: Within 5 min of infusion, IGF-1 had spread from the infused lateral ventricle into and through the third and fourth ventricles. At this time, 25% of the infused IGF-1 had disappeared from the CSF-brain-meningeal system; the half time of this loss was 12 min. The plasma concentration of cleared IGF-1 was, however, very low from 2 to 9 min and only began to rise markedly after 20 min. This delay between loss and gain plus the lack of radiotracer in the cortical subarachnoid space suggested that much of the IGF-1 was cleared into blood via the cranial and/or spinal nerve roots and their associated lymphatic systems rather than periventricular tissue and arachnoid villi. Less than 10% of the injected radioactivity remained in the CSF-brain system after 180 min. The CSF and arteries and arterioles within the subarachnoid cisterns were labeled with IGF-1 within 10 min. Between 60 and 180 min, most of the radioactivity within the cranium was retained within and around these blood vessels and by periaqueductal gray matter. Tissue profiles at two sites next to ventricular CSF showed that IGF 1 penetrated less than 1.25 mm into brain tissue and appreciable 125I-activity remained at the tissue-ventricular CSF interface after 180 min. CONCLUSION: Our findings suggest that entry of IGF-1 into normal brain parenchyma after lateral ventricle administration is limited by rapid clearance from CSF and brain and slow movement, apparently by diffusion, into the periventricular tissue. Various growth factors and other neuroactive agents have been reported to be neuroprotective within the injured brain after intraventricular administration. It is postulated that the delivery of such factors to neurons and glia in the injured brain may be facilitated by abnormal CSF flow. These several observations suggest that the flow of CSF and entrained solutes may differ considerably between normal and abnormal brain and even among various neuropathologies. PMID- 16045807 TI - Biomechanics of bone-fracture fixation by stiffness-graded plates in comparison with stainless-steel plates. AB - BACKGROUND: In the internal fixation of fractured bone by means of bone-plates fastened to the bone on its tensile surface, an on-going concern has been the excessive stress-shielding of the bone by the excessively-stiff stainless-steel plate. The compressive stress-shielding at the fracture-interface immediately after fracture-fixation delays callus formation and bone healing. Likewise, the tensile stress-shielding of the layer of the bone underneath the plate can cause osteoporosis and decrease in tensile strength of this layer. METHOD: In order to address this problem, we propose to use stiffness-graded plates. Accordingly, we have computed (by finite-element analysis) the stress distribution in the fractured bone fixed by composite plates, whose stiffness is graded both longitudinally and transversely. RESULTS: It can be seen that the stiffness graded composite-plates cause less stress-shielding (as an example: at 50% of the healing stage, stress at the fracture interface is compressive in nature i.e. 0.002 GPa for stainless steel plate whereas stiffness graded plates provides tensile stress of 0.002 GPa. This means that stiffness graded plate is allowing the 50% healed bone to participate in loadings). Stiffness-graded plates are more flexible, and hence permit more bending of the fractured bone. This results in higher compressive stresses induced at the fractured faces accelerate bone healing. On the other hand, away from the fracture interface the reduced stiffness and elastic modulus of the plate causes the neutral axis of the composite structure to be lowered into the bone resulting in the higher tensile stress in the bone-layer underneath the plate, wherein is conducive to the bone preserving its tensile strength. CONCLUSION: Stiffness graded plates (with in built variable stiffness) are deemed to offer less stress-shielding to the bone, providing higher compressive stress at the fractured interface (to induce accelerated healing) as well as higher tensile stress in the intact portion of the bone (to prevent bone remodeling and osteoporosis). PMID- 16045808 TI - Evaluating teaching effectiveness in nursing education: an Iranian perspective. AB - BACKGROUND: The main objective of this study was to determine the perceptions of Iranian nurse educators and students regarding the evaluation of teaching effectiveness in university-based programs. METHODS: An exploratory descriptive design was employed. 143 nurse educators in nursing faculties from the three universities in Tehran, 40 undergraduate, and 30 graduate students from Tehran University composed the study sample. In addition, deans from the three nursing faculties were interviewed. A researcher-developed questionnaire was used to determine the perceptions of both faculty and students about evaluating the teaching effectiveness of nurse educators, and an interview guide was employed to elicit the views of deans of faculties of nursing regarding evaluation policies and procedures. Data were analyzed using parametric and nonparametric statistics to identify similarities and differences in perceptions within the Iranian nurse educator group and the student group, and between these two groups of respondents. RESULTS: While faculty evaluation has always been a major part of university based nursing programs, faculty evaluation must be approached more analytically, objectively, and comprehensively to ensure that all nursing educators receive the fairest treatment possible and that the teaching-learning process is enhanced. CONCLUSION: Educators and students stressed that systematic and continuous evaluation as well as staff development should be the primary goals for the faculty evaluation process. The ultimate goals is the improvement of teaching by nurse educators. PMID- 16045809 TI - Overexpression of cathepsin K during silica-induced lung fibrosis and control by TGF-beta. AB - BACKGROUND: Lung fibrosis is characterized by tissue remodeling resulting from an imbalance between synthesis and degradation of extracellular organic matrices. To examine whether cathepsin(s) (Cat) are important in the development of pulmonary fibrosis, we assessed the expression of four Cat known for their collagenolytic activity in a model of silica-induced lung fibrosis. METHODS: Different strains of mice were transorally instilled with 2.5 mg crystalline silica or other particles. Cat expression (Cat K, S, L and B) was quantified in lung tissue and isolated pulmonary cells by quantitative RT-PCR. In vitro, we assessed the effect of different cytokines, involved in lung inflammatory and fibrotic responses, on the expression of Cat K by alveolar macrophages and fibroblasts. RESULTS: In lung tissue, Cat K transcript was the most strongly upregulated in response to silica, and this upregulation was intimately related to the fibrotic process. In mouse strains known for their differential response to silica, we showed that the level of Cat K expression following silica treatment was inversely related to the level of TGF-beta expression and the susceptibility of these strains to develop fibrosis. Pulmonary macrophages and fibroblasts were identified as Cat K overproducing cells in the lung of silicotic mice. In vitro, Cat K was downregulated in mouse and human lung fibroblasts by the profibrotic growth factor TGF-beta1. CONCLUSION: Altogether, these data suggest that while Cat K may contribute to control lung fibrosis, TGF-beta appears to limit its overexpression in response to silica particles. PMID- 16045810 TI - Randomized controlled trial of the cognitive side-effects of magnetic seizure therapy (MST) and electroconvulsive shock (ECS). AB - Magnetic seizure therapy (MST) is under development as a means of improving the cognitive side-effect profile of electroconvulsive therapy (ECT) by inducing more spatially delimited seizures that spare cortical regions involved in memory. We tested whether MST had a cognitive side-effect profile distinct from electroconvulsive shock (ECS) in a non-human primate model, using the Columbia University Primate Cognitive Profile, which has been shown to be sensitive to the cognitive effects of ECS. Using a within-subject cross-over design, daily ECS, MST, and sham (anaesthesia-only) interventions were administered in 5-wk blocks. Rhesus macaques (n = 2) were trained on a long-term memory task, an anterograde learning and memory task, and a combined anterograde and retrograde task where learning and memory were evaluated for new and previously learned 3-item lists. Acutely following each intervention, monkeys were tested on the cognitive battery twice daily, separated by a 3-h retention interval. Overall, monkeys were least accurate following ECS (p's < 0.05) compared to sham and MST. This effect was most marked for long-term memory of a constant target, short-term memory of a variable target and recall of previously learned 3-item lists. Monkeys were slowest to complete all tasks following ECS (p's = 0.0001). Time to task completion following MST did not differ from sham. These findings suggest that MST results in a more benign acute cognitive side-effect profile than ECS in this model, consistent with initial observations with human MST. PMID- 16045811 TI - The interaction of versican with its binding partners. AB - Versican belongs to the family of the large aggregating chondroitin sulfate proteoglycans located primarily within the extracellular matrix (ECM). Versican, like other members of its family, has unique N- and C-terminal globular regions, each with multiple motifs. A large glycosaminoglycan-binding region lies between them. This review will begin by outlining these structures, in the context of ECM proteoglycans. The diverse binding partners afforded to versican by virtue of its modular design will then be examined. These include ECM components, such as hyaluronan, type I collagen, tenascin-R, fibulin-1, and -2, fibrillin-1, fibronectin, P- and L-selectins, and chemokines. Versican also binds to the cell surface proteins CD44, integrin beta 1, epidermal growth factor receptor, and P selectin glycoprotein ligand-1. These multiple interactors play important roles in cell behaviour, and the roles of versican in modulating such processes are discussed. PMID- 16045812 TI - Protein kinase clk/STY is differentially regulated during erythroleukemia cell differentiation: a bias toward the skipped splice variant characterizes postcommitment stages. AB - Clk/STY is a LAMMER protein kinase capable to phosphorylate serine/arginine-rich (SR) proteins that modulate pre-mRNA splicing. Clk/STY alternative splicing generates transcripts encoding a full-length kinase and a truncated catalytically inactive protein. Here we showed that clk/STY, as well as other members of the family (e.g. clk2, clk3 and clk4), are up-regulated during HMBA-induced erythroleukemia cell differentiation. mRNAs coding for the full-length and the truncated forms were responsible for the overall increased expression. In clk/STY, however, a switch was observed for the ratio of the two alternative spliced products. In undifferentiated cells the full-length transcript was more abundant whereas the transcript encoding for the truncated form predominated at latter stages of differentiation. Surprisingly, overexpression of clk/STY did not alter the splicing switch upon differentiation in MEL cells. These results suggest that clk/STY might contribute to control erythroid differentiation by a mechanism that implicates a balance between these two isoforms. PMID- 16045813 TI - Identification of novel nuclear localization signal within the ErbB-2 protein. AB - ErbB2, a member of the receptor tyrosine kinase family, is frequently over expressed in breast cancer. Proteolysis of the extracellular domain of ErbB2 results in constitutive activation of ErbB2 kinase. Recent study reported that ErbB2 is found in the nucleus. Here, we showed that ErbB2 is imported into the nucleus through a nuclear localization signal (NLS)-mediated mechanism. The NLS sequence KRRQQKIRKYTMRR (aa655-668) contains three clusters of basic amino acids and it is sufficient to target GFP into the nucleus. However, mutation in any basic amino acid cluster of this NLS sequence significantly affects its nuclear localization. Furthermore, it was found that this NLS is essential for the nuclear localization of ErbB2 since the intracellular domain of Erb2 lacking NLS completely abrogates its nuclear translocation. Taken together, our study identified a novel nuclear localization signal and reveals a novel mechanism underlying ErbB2 nuclear trafficking and localization. PMID- 16045814 TI - Gene expression alteration during redox-dependent enhancement of arsenic cytotoxicity by emodin in HeLa cells. AB - Emodin (1,3,8-trihydroxy-6-methylanthraquinone) could enhance the sensitivity of tumor cells to arsenic trioxide (As2O3)-induced apoptosis via generation of ROS, but the molecular mechanism has not been elucidated. Here, we carried out cDNA microarray-based global transcription profiling of HeLa cells in response to As2O3/emodin cotreatment, comparing with As2O3-only treatment. The results showed that the expression of a number of genes was substantially altered at two time points. These genes are involved in different aspects of cell function. In addition to redox regulation and apoptosis, ROS affect genes encoding proteins associated with cell signaling, organelle functions, cell cycle, cytoskeleton, etc. These data suggest that based on the cytotoxicity of As2O3, emodin mobilize every genomic resource through which the As2O3-induced apoptosis is facilitated. PMID- 16045815 TI - Induction of HLA-G expression in a melanoma cell line OCM-1A following the treatment with 5-aza-2'-deoxycytidine. AB - The non-classical HLA class I antigen HLA-G is an immune modulator which inhibits the functions of T cells, NK cells, and the Dendritic cells (DC). As a result, HLA-G expression in malignant cells may provide them with a mechanism to escape the immune surveillance. In melanoma, HLA-G antigen expression has been found in 30% of surgically removed lesions but in less than 1% of established cell lines. One possible mechanism underlying the differential HLA-G expression in vivo and in vitro is that the HLA-G gene is epigenetically repressed in melanoma cells in vitro. To test this hypothesis, we treated the HLA-G negative melanoma cell line OCM-1A with the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (5-AC) and analyzed whether HLA-G expression can be restored. Our data strongly suggest that HLA-G is silenced as a result of CpG hypermethylation within a 5' regulatory region encompassing 220 bp upstream of the start codon. After treatment, HLA-G mRNA expression was dramatically increased. Western blot and flow cytometry showed that HLA-G protein was induced. Interestingly, HLA-G cell surface expression on the 5-AC treated OCM-1A cells is much less than that on the HLA-G positive JEG-3 cells while a similar amount of total HLA-G was observed. Possible mechanisms for the difference were analyzed in the study such as cell cold treatment, peptide loading and antigen processing machinery components (APM) as well as beta2 microglobulin (beta2-m) expression. Data revealed that the APM component calreticulin might be involved in the lower HLA-G surface expression on OCM-1A cells. Taken together, our results indicated that DNA methylation is an important epigenetic mechanism by which HLA-G antigen expression is modulated in melanoma cells in vitro. Furthermore, to the first time, we hypothesized that the deficiency of calreticulin might be involved in the low HLA-G surface expression on the 5-AC treated OCM-1A cells. PMID- 16045816 TI - Ultrastructure and histochemistry of rat myocardial capillary endothelial cells in response to diabetes and hypertension. AB - Insufficient growth and rarefaction of capillaries, followed by endothelial dysfunction may represent one of the most critical mechanisms involved in heart damage. In this study we examined histochemical and ultrastructural changes in myocardial capillary endothelium in two models of heart failure streptozotocin induced diabetes mellitus (STZ) and NO-deficient hypertension in male Wistar rats. Diabetes was induced by a single i.v. dose of STZ (45 mg/kg) and chronic 9 week stage was analysed. To induce NO-deficient hypertension, animals were treated with inhibitor of NO synthase L-nitroarginine methylester (L-NAME) (40 mg/kg) for 4 weeks. Left ventricular tissue was processed for enzyme catalytic histochemistry of capillary alkaline phosphatase (AlPh), dipeptidyl peptidase IV (DPP IV), and endothelial NO synthase/NADPH-diaphorase (NOS) and for ultrastructural analysis. In diabetic and hypertensive rats, lower/absent AlPh and DPP IV activities were found in focal micro-areas. NOS activity was significantly reduced and persisted only locally. Quantitative evaluation demonstrated reduction of reaction product intensity of AlPh, DPP and NOS by 49.50%, 74.36%, 20.05% in diabetic and 62.93%, 82.71%, 37.65% in hypertensive rats. Subcellular alterations of endothelial cells were found in heart of both groups suggesting injury of capillary function as well as compensatory processes. Endothelial injury was more significant in diabetic animals, in contrast the adaptation was more evident in hypertensive ones. CONCLUDING: both STZ-induced diabetes- and NO-deficient hypertension-related cardiomyopathy were accompanied by similar features of structural remodelling of cardiac capillary network manifested as angiogenesis and angiopathy. The latter was however, predominant and may accelerate disappearance of capillary endothelium contributing to myocardial dysfunction. PMID- 16045817 TI - Mesenchymal stem cells derived from human placenta suppress allogeneic umbilical cord blood lymphocyte proliferation. AB - Human placenta-derived mononuclear cells (MNC) were isolated by a Percoll density gradient and cultured in mesenchymal stem cell (MSC) maintenance medium. The homogenous layer of adherent cells exhibited a typical fibroblast-like morphology, a large expansive potential, and cell cycle characteristics including a subset of quiescent cells. In vitro differentiation assays showed the tripotential differentiation capacity of these cells toward adipogenic, osteogenic and chondrogenic lineages. Flow cytometry analyses and immunocytochemistry stain showed that placental MSC was a homogeneous cell population devoid of hematopoietic cells, which uniformly expressed CD29, CD44, CD73, CD105, CD166, laminin, fibronectin and vimentin while being negative for expression of CD31, CD34, CD45 and alpha-smooth muscle actin. Most importantly, immuno-phenotypic analyses demonstrated that these cells expressed class I major histocompatibility complex (MHC-I), but they did not express MHC-II molecules. Additionally these cells could suppress umbilical cord blood (UCB) lymphocytes proliferation induced by cellular or nonspecific mitogenic stimuli. This strongly implies that they may have potential application in allograft transplantation. Since placenta and UCB are homogeneous, the MSC derived from human placenta can be transplanted combined with hematopoietic stem cells (HSC) from UCB to reduce the potential graft-versus-host disease (GVHD) in recipients. PMID- 16045818 TI - Calmodulin regulates the post-anaphase reposition of centrioles during cytokinesis. AB - A transient postanaphase repositioning of the centriole is found to control the completion of cytokinesis. Using a green fluorescent protein-calmodulin fusion protein as a living cell probe, we have previously found that calmodulin is associated with the initiation and progression of cytokinesis. In this study, we further studied the effect of calmodulin on the repositioning of the centriole and subsequent cell cycle progression. When activity of calmodulin is inhibited, the regression of the centriole from the intercellular bridge to the cell center is blocked, and thus the completion of cell division is repressed and two daughter cells are linked by longer cell bridge in perturbed cells. W7 treatment during cytokinesis also results in unfinished cytokinesis and stopped G1 phase. These results suggest that calmodulin activity is required for centriole repositioning and can affect the completion of cytokinesis and cell cycle progression. PMID- 16045819 TI - The Wizard of Oz...if he only had stem cells! PMID- 16045820 TI - Wage increases in the clinical laboratory: how are workers faring against inflation and in comparison to other health professions? AB - High vacancy rates in the clinical laboratory profession have led to the use of wage increases and financial incentives to attract and retain workers. American Society for Clinical Pathology (ASCP) surveys indicate that wages for Medical Technologists and Medical Laboratory Technicians have been steadily rising in the past few years following years of little or no increases. When adjusted for inflation, the real wage increases have even modestly exceeded the inflation rate. However, wages in the clinical laboratory remain lower than in several other allied health professions with comparable educational preparation. Achieving competitive wages will be important in addressing the long-term need to attract more students to the clinical laboratory. PMID- 16045821 TI - Recentralizing phlebotomy back into the laboratory. AB - In many hospitals, the responsibility for performing phlebotomy has shifted from the laboratory to the nursing unit. However, few nursing programs have incorporated phlebotomy instruction into their employee training. As a result, organizations have experienced varying degrees of success with this shift. This article shares the experiences of one hospital that recentralized phlebotomy duties one year ago, including information on data collection, financial analysis, and training orientation. PMID- 16045822 TI - Proficiency testing protocol. PMID- 16045823 TI - The medical review officer and conflicts of interest. PMID- 16045824 TI - Quality and personnel competency. PMID- 16045827 TI - Clinical trials in giant cell arteritis. PMID- 16045826 TI - TNF-blocking therapy in rheumatoid arthritis and ankylosing spondylitis: why is cost-effectiveness a major issue? PMID- 16045825 TI - West meets East in rheumatology. PMID- 16045828 TI - Classification of the vasculitides: are they clinically useful? AB - Several classification systems for the vasculitides have been developed over the second half of the past century. These classifications systems have been modified according to the advances made in the differentiation of these syndromes, particularly their histopathologic features (the vessel size involved) and their clinical features. Although these classifications have been widely used in the clinical setting for the diagnosis of these disorders, they were developed for research purposes. Given the common clinical and, sometimes, histopathologic features of the different vasculitides, a classification system that facilitates a diagnosis and the formulation of a therapeutic plan is still needed. The development and utilization of novel serum markers and new imaging modalities coupled with advances in the understanding of these disorders, may allow the development of a classification system for the vasculitides that will be of greater value in the clinical setting. PMID- 16045830 TI - Cutaneous vasculitis. AB - Vasculitis is defined as inflammation of blood vessels and can affect multiple organs. Several classification systems exist to categorize vasculitis such as vessel size, presence of anti-neutrophil cytoplasmic antibody, pathogenesis of the inflammation, and type of inflammatory cell infiltrate. Cutaneous vasculitis occurs as a manifestation of many diseases including rheumatologic diseases, hypersensitivity syndromes, infections, and malignancies. The diagnosis of the cutaneous vasculitis and the underlying cause requires a complete history and physical exam and usually a biopsy or angiogram. The treatment depends on the etiology of the inflammation and includes immunosuppression, withdrawal of the offending agent, antibacterial/antiviral agents, and chemotherapies. A clear understanding and approach to this condition will improve the physician's ability to provide optimal patient care. PMID- 16045829 TI - Epidemiology of systemic vasculitis. AB - The systemic vasculitides are heterogeneous conditions of unknown etiology characterized by inflammation and necrosis of different sized blood vessels. Wegener's Granulomatosis, microscopic polyangiitis, and Churg Strauss syndrome are associated with anti-neutrophil cytoplasmic antibodies and affect small and medium blood vessels. They are very rare in childhood and peak in the 65 to 70 year old age group. Wegener's Granulomatosis appears to be more common in the North of Europe compared with the South. All are more common in whites compared with other populations. Genetic and environmental factors, including infection, drugs, and silica, are important in etiology. Giant cell arteritis is predominantly a disease of whites over the age of 50. It appears more common in individuals with Nordic descent. Incidence may be increasing over time and cyclical variation in disease may reflect an infectious etiology. Takayasu arteritis is a disease of the aorta and its branches, however pulmonary and cardiac arteries may be involved. Patients are usually under 40-years of age at presentation and there are no apparent differences in incidence or clinical characteristics/aortic involvement across the globe. Kawasaki disease (KD) and Henoch-Schonlein purpura are diseases of children and rarely affect adults. Both have been reported to be more common in Asians than whites. The incidence of KD is higher in Japan and China compared with other regions. No definite trigger factors have been found, but KD has been linked to infection, house dust mite and chemicals, and Henoch-Schonlein purpura to a pesticide and drugs. PMID- 16045831 TI - Infection-related vasculitis. AB - Infection-related vasculitis constitutes the most common cause of secondary vasculitis. A great variety of microorganisms can induce directly or indirectly inflammatory vascular damage resulting in vascular occlusion, tissue ischemia, and necrosis. In the developed world hepatitis B and C-related vasculitis remain the most common clinical syndromes, while HIV-associated vasculitis remains a concern in developing countries. PMID- 16045832 TI - Polyarteritis nodosa revisited. AB - Polyarteritis nodosa (PAN), the prototype of systemic vasculitis, is a rare condition characterized by necrotizing inflammation of medium-sized or small arteries without glomerulonephritis or vasculitis in arterioles, capillaries, or venules. Signs and symptoms of this disease are primarily attributable to diffuse vascular inflammation and ischemia of affected organs. Virtually any organ with the exception of the lungs may be affected, with peripheral neuropathy and symptoms from osteoarticular, renal artery, and gastrointestinal tract involvement being the most frequent clinical manifestations. A clear distinction between limited versus systemic disease and idiopathic versus hepatitis B related PAN should be done because there are differences in the implicated pathogenetic mechanisms, their treatment, and prognosis. Currently, corticosteroids plus cyclophosphamide is the standard of care for idiopathic PAN, in particular for patients with adverse prognostic factors (more severe disease), in whom this combination prolonged survival. In contrast for hepatitis B related PAN treatment consists of schemes that include plasmapheresis and antiviral agents. PMID- 16045833 TI - Outcome measures in psoriatic arthritis clinical trials. AB - Outcome measures in psoriatic arthritis (PsA) have been primarily borrowed from the assessment of rheumatoid arthritis and ankylosing spondylitis, although several specific measures for PsA have been established. The advent of new therapeutic agents for the treatment of PsA has made the need for specific outcome measures for PsA more critical to evaluate the heterogeneous manifestations of this disease and features that are unique to its assessment. Several outcome measures have been validated for use in PsA clinical trials while others are being evaluated by groups such as The Group for Assessment of Psoriasis and Psoriatic Arthritis for future use in clinical trials. PMID- 16045834 TI - Genetic factors in psoriatic arthritis. AB - The genetic factors that are associated with psoriatic arthritis (PsA) are intricately linked with those that predispose to psoriasis itself. The strongest association is with human leukocyte antigen-Cw*0602, although true susceptibility may lie with one of the neighboring genes along a disease-associated haplotype. There are a number of interesting candidate genes within the major histocompatibility complex (MHC) region with strong functional relevance that have been investigated in PsA. In addition, several areas outside the MHC complex have been highlighted as a result of genetic linkage studies in psoriasis. PsA is a complex, multifactorial disease where multiple genes are likely to influence disease susceptibility, severity, and clinical phenotype. The current evidence for genetic factors in psoriasis and PsA will be reviewed. PMID- 16045835 TI - Treatment advances in psoriatic arthritis. AB - Treatment of psoriatic arthritis, like the treatment of rheumatoid arthritis, now commonly includes the use of inhibitors of tumor necrosis factor in addition to traditional synthetic disease-modifying antirheumatic drugs. This paper examines the most recent data from therapeutic trials in psoriatic arthritis, with particular emphasis on the effectiveness of the tumor necrosis factor inhibitors. Recent data on potential future therapies is discussed as well, along with data on the mechanisms of current therapies that may have relevance for future treatment approaches. PMID- 16045836 TI - The role of ultrasound in the diagnosis and management of psoriatic arthritis. AB - Psoriatic arthritis (PsA) presents many diagnostic, management and research challenges for rheumatologists who wish to obtain early diagnosis, differentiate synovitis and enthesitis, monitor disease activity accurately and objectively, prevent the development of structural damage, deliver local therapy accurately, and obtain PsA tissue for research purposes. Musculoskeletal ultrasound (MSUS) is widely used by European rheumatologists in their clinical practice to meet these challenges and has the potential to become the rheumatologist's stethoscope in Europe and North America. This paper examines the evidence that MSUS can improve clinical evaluation of patients with PsA for synovitis and enthesitis, that MSUS is more sensitive than plain radiography in detecting structural damage in joints, that MSUS can improve the success of joint aspiration and guide biopsy of PsA tissues. Recent exciting developments in the management of PsA are detailed including the role of power Doppler in the diagnosis of enthesitis in PsA, the role of MSUS in objective monitoring of disease activity, the evaluation of MSUS in the diagnosis of sacroiliitis, and the use of MSUS to guide therapeutic injection of the sacroiliac joints. PMID- 16045837 TI - Angiogenesis in psoriasis and psoriatic arthritis: clues to disease pathogenesis. AB - Psoriasis is a common chronic dermatosis occurring in 2% of the population and associated with an inflammatory arthritis--psoriatic arthritis (PsA)--in up to 40% of cases. PsA accounts for approximately 15% of patients attending early synovitis clinics, therefore it represents the second most common diagnostic category after rheumatoid arthritis. There are a number of common pathogenic features that link the skin and the joint inflammatory processes. Angiogenesis appears to be a fundamental inflammatory response early in the pathogenesis and significant abnormalities of vascular morphology and angiogenic growth factors have been described in psoriasis and PsA. This paper will explore the recent published evidence to support the hypothesis that dysregulated angiogenesis provides a primary pathogenic mechanism in psoriasis skin and in the PsA joint. PMID- 16045838 TI - Dorsal modified calcaneal plate for extensive midfoot arthrodesis. AB - BACKGROUND: Extensive midfoot fusions can be challenging because of bone loss, deformity, and soft tissue anatomy. Several options have been advocated, including multiple screw fixation, medial plating, and plantar plating. We report a new technique using a dorsally-placed, modified calcaneal plate for treatment of this difficult clinical problem. METHODS: Patients undergoing extensive (more than four joints) midfoot arthrodeses with a dorsally-placed, modified calcaneal plate between 2000 and 2003 were retrospectively reviewed. Diagnoses included Charcot arthropathy (four), osteoarthritis (two), posttraumatic osteoarthritis (two), massive bone loss from previous infection (one), and residual clubfoot deformity (one). Patients with active midfoot infections were excluded. During the study period, midfoot arthrodeses with a dorsal calcaneal plate were done in 10 patients. Of these, nine patients were available for review. Arthrodeses were attempted in 62 joints in these nine patients. Autogenous grafting was used in three patients (23 joints), allograft was used in six patients (39 joints). Patients were maintained nonweightbearing until radiographs or computed tomography conclusively showed union. RESULTS: One of the 10 patients died from an unrelated cause. In the nine remaining patents, 95% (59 of 62) of joints fused within 4 months of surgery. Postoperative complications included nonunion with broken screws in one patient, and three wound problems successfully treated with local dressings. Secondary procedures included one revision arthrodesis and two hardware removals. Patient satisfaction with this procedure was very high (eight of nine). CONCLUSIONS: The use of a dorsal calcaneal plate is a viable method of fixation for achieving fusion in extensive midfoot arthropathy. The plate is low profile and easily moldable to conform to dorsal midfoot anatomy. It can be placed without extensive plantar or medial foot dissection and maintains midfoot alignment until bony fusion occurs. In patients with complex midfoot pathology requiring multijoint fusions, the results have been satisfactory. PMID- 16045839 TI - Combination of hook plate and tibial pro-fibular screw fixation of osteoporotic fractures: a clinical evaluation of operative strategy. AB - BACKGROUND: Internal fixation of osteoporotic ankle fractures is technically difficult and may fail because of unreliable purchase. This study was undertaken to determine if a combination of a hook plate and tibial pro-fibular screws can provide secure fixation until fracture union. METHODS: Thirty-one patients between the ages of 55 and 90 years had open reduction and internal fixation of ankle fractures between April, 2001, and April, 2003. Sixteen patients with an average age of 71.4 years had ankle fracture fixation with a combination of hook plate and tibial pro-fibular screws for the distal fibular fracture, and 15 patients with an average age of 71.9 years had fixation of their ankle fractures with standard fixation technique using AO/ASIF principles but no tibial pro fibular screws. All patients were followed with clinical and radiologic assessment at 2 weeks, 6 weeks, and 12 weeks postoperatively. At an average of 15.8 months after injury, patients also completed a mailed questionnaire with the Olerud-Molander ankle score and the AOFAS ankle-hindfoot score for preoperative and postoperative status. RESULTS: All patients who had tibial pro-fibular screw fixation had fracture union without hardware failure or complications. In the standard fixation group two patients had wound breakdown and one had a valgus malunion with screw pull out. The AOFAS and Olerud-Molander scores for the standard open reduction and internal fixation were 57.3 and 82.8 before injury and 37 and 43.8 postoperatively, respectively. The AOFAS and Olerud-Molander scores for the hook plate and tibial pro-fibular fixation group were 55.9 and 81.3 before injury and 42.4 and 50.3 postoperatively, respectively. CONCLUSIONS: The combination of hook plate and tibial pro-fibular screws in osteoporotic ankle fractures in a series of patients has not been reported before. This novel technique provides stable fixation for osteoporotic ankle fractures in elderly patients until union is achieved with good clinical scores. PMID- 16045840 TI - Bone suture anchor fixation in the lower extremity: a review of insertion principles and a comparative biomechanical evaluation. AB - BACKGROUND: Suture anchors have been developed for the fixation of ligaments, capsules, or tendons to bone. These devices have led to improved fixation, smaller incisions, earlier limb mobility, and improved outcomes. They were originally developed for use in shoulder reconstructions but are now used in almost all extremities. In the lower leg they are used in the tibia, the talus, the calcaneus, tarsal bones, and phalanges. Nevertheless, techniques for insertion and mechanisms of failure are not well described. METHODS: Five suture anchors were studied to determine the pullout strength in four distal cadaver femurs and four proximal cadaver tibias from 55- and 62-year-old males. Eight hundred ninety Newton line was used, testing the anchors to failure with an Instron testing device (Instron, Norwood, MA). The anchor devices were inserted randomly and tested blindly (12 tests per anchor device, 60 tests in all). RESULTS: Two anchors in each group tested failed at low loads. Both types of plastic anchors had failures at the eyelet. Average pullout strength varied from 85.4 to 185.6 N. CONCLUSIONS: Insertion techniques are specific for each device, and they must be followed for optimal fixation. In this study, in all five groups of anchors tested two of the 12 anchors in each group failed with minimal force. On the basis of this finding we recommend that, if suture anchor fixation is necessary, at least two anchors should be used. Since there appears to be a percentage of failure in all devices, the second anchor can serve as a backup. It is imperative that surgeons be familiar with the insertion techniques of each device before use. PMID- 16045841 TI - Plantar pressures in the forefoot after lateral column lengthening: a cadaver study comparing the Evans osteotomy and calcaneocuboid fusion. AB - BACKGROUND: Excessive varus alignment of the forefoot after lateral column lengthening has been reported to lead to overloading of the lateral foot. The purpose of this study was to investigate whether there is a difference between the Evans opening wedge calcaneal osteotomy (Evans) and the calcaneocuboid distraction arthrodesis (CCDA) with respect to lateral forefoot loading. METHODS: In each of 12 matched pairs of cadaver feet, plantar pressure measurements of the intact specimens were obtained during simulated foot-flat and early heel-rise phases of gait and again after randomly performing the Evans procedure on one foot and the CCDA on the other foot. Cervical plate fixation was used for immediate stability. RESULTS: Both procedures resulted in statistically significant increased loading of the lateral forefoot and decreased loading of the medial forefoot compared with the preoperative status. The relative increase in lateral pressures was statistically greater with the CCDA than with the Evans. The average increase in pressure under the fifth metatarsal head in the foot-flat phase was 46% +/- 42% (range-4% to 141%) with the Evans and 104% +/- 58% (range 9% to 216%) with the CCDA (p = 0.003). In the early heel-rise phase, the increase in pressure was 50% +/- 43% (range 2% to 108%) and 96% +/- 65% (range 12% to 263%), respectively (p = 0.02). CONCLUSION: The experimental data suggest that lateral column overload may be more likely with the CCDA than with the Evans. Physicians should be aware of the likelihood of increasing lateral column loads with both the CCDA and the Evans procedure. It may be possible to avoid this problem by using less lateral column lengthening than the 1 cm used in this study or by considering a medial column arthrodesis or plantarflexion osteotomy to balance forefoot loading. PMID- 16045842 TI - Blood flow to the metatarsal head after chevron bunionectomy. AB - BACKGROUND: Avascular necrosis (AVN) of the metatarsal head is a known, but infrequent, complication of a chevron bunionectomy. An increased incidence of AVN has been reported with the addition of an adductor tenotomy and lateral release. The purpose of this study was to document the vascular insult to the metatarsal head during a chevron bunionectomy with adductor tenotomy and lateral release through direct blood flow measurements during the procedure. METHODS: A prospective study of 20 patients was performed. A chevron bunionectomy was performed under regional anesthesia without tourniquet control by two surgeons. Blood flow recordings were documented during key portions of the procedure: baseline, medial capsulotomy, adductor tenotomy and lateral release, and chevron osteotomy. The Periflux PF3 laser Doppler probe (Perimed, Inc., North Royalton, OH) was used to record blood flow. Mauchly's test of Sphericity and a pairwise comparison were performed with significance defined as p < 0.05. All patients had radiographs 3 months postoperatively to assess AVN. RESULTS: There was a statistically significant decrease in blood flow to the metatarsal head at each portion of the procedure. The greatest insult occurred with the medial capsulotomy (45% decrease). The lateral release and adductor tenotomy caused a 13% decrease and the chevron osteotomy caused a 13% decrease, totaling 71% decrease from the baseline. There was no evidence of AVN at 3 months, and all patients had radiographic evidence of union without recurrence or over correction. CONCLUSION: The addition of the adductor tenotomy with lateral release often is crucial to obtain the desired correction and avoid the complication of recurrence. The disruption of microvascular blood flow to the metatarsal head after a chevron bunionectomy with adductor tenotomy and lateral release was not complete, totaling 71% in the study population. The greatest insult occurred with the medial capsulotomy. There were no occurrences of AVN. Despite the fear of AVN, this study demonstrated that an adductor tenotomy and lateral release could be done safely completed with a chevron bunionectomy. PMID- 16045843 TI - Tibiotalocalcaneal arthrodesis: a biomechanical assessment of stability. AB - BACKGROUND: Combined ankle and subtalar (tibiotalocalcaneal) arthrodesis is a procedure that can be used to successfully treat disabling foot and ankle arthropathy and is a reasonable salvage alternative to amputation for the treatment of nonbraceable neuropathic, diabetic, degenerative, or rheumatoid joints. Although many methods of tibiotalocalcaneal (TTC) arthrodesis have been described in the literature, the most popular current methods involve the use of crossed cancellous bone screws, plates, or a locked retrograde intramedullary rod. Fusion in these patients can be difficult, with significant complications including infection, malunion, and nonunion. A persistent nonunion can lead to failure of the hardware and recurrent deformity. METHODS: We biomechanically tested the stability and micromotion in four methods of TTC arthrodesis using liquid metal strain gauges and Instron (Norwood, MA) material testing systems. Anatomically identical synthetic bones with properties very similar to human bone were instrumented and tested. Four instrumentation techniques were tested: 1) three crossed 6.5-mm cancellous screws, 2) two crossed 6.5-mm cancellous screws, 3) locked retrograde intramedullary rod, and 4) locked retrograde intramedullary rod augmented with a single anteromedial bone staple. Six separate specimens for each technique were tested. RESULTS: The three crossed cancellous screw technique provided the greatest stability with respect to micromotion (p < 0.05). The addition of a tibiotalar staple to the locked intramedullary rod conferred stability nearly equal to that of the three crossed cancellous screw fixation (p < 0.05). The locked intramedullary rod group and the two crossed cancellous screw group allowed significant micromotion at the arthrodesis sites, which was a full order of magnitude higher (p < 0.05) than in the three crossed cancellous screw group and the staple augmented intramedullary rod group. CONCLUSIONS: Biomechanically, a staple augmented locked intramedullary rod for TTC arthrodesis confers excellent stability nearly equal to the three crossed cancellous screw technique for TTC arthrodesis. PMID- 16045844 TI - Functional limitations of patients with end-stage ankle arthrosis. AB - BACKGROUND: Arthritis and other rheumatic conditions are the leading causes of disability among adults in the United States. The purpose of this report was to describe the self-reported functional limitations of a group of patients with end stage ankle arthrosis. METHOD: Patients who presented for operative management of end-stage ankle arthrosis at the University of Minnesota and Harborview Medical Center completed a Musculoskeletal Functional Assessment (MFA) as part of their preoperative clinical evaluation. Data from patients evaluated during the time period April, 1995, through May, 2004, were used for this project. RESULTS: Four hundred and twenty-six patients with the diagnosis of end-stage ankle arthrosis completed baseline questionnaires. Six of the 426 patients received care on both ankles during the time of this project. The average age of patients at the time of completion of the questionnaire was 56.7 years. There were 241 men and 185 women. The primary underlying causes identified by the treating surgeon at the time of surgery were primary osteoarthritis with no known prior trauma (66), previous trauma (tibial fracture, foot fractures, or ankle ligamentous disruption) (296), rheumatoid arthritis (24), no known cause (21), and a variety of diseases or infections (19). In all domains, the patients with end-stage ankle arthrosis showed statistically significant differences from a general population sample. CONCLUSIONS: The effects of ankle arthritis as demonstrated by this data are severe. Most of these patients were severely limited in function. Without a data-driven understanding of the limitations the patients have, it is difficult to make an effective argument for focused research to solve the problems. Without understanding the patients' needs, it is impossible to assess the effect of treatment. The information in this paper provides a baseline understanding of effect of the current functional limitations of patients with end-stage ankle arthrosis. PMID- 16045845 TI - Quantification of muscle strength and imbalance in neurogenic pes cavus, compared to health controls, using hand-held dynamometry. AB - BACKGROUND: Pes cavus foot deformity in neuromuscular disease is thought to be related to an imbalance of musculature around the foot and ankle. The most common cause of neurogenic pes cavus is Charcot-Marie-Tooth (CMT) disease. The aim of this investigation was to objectively quantify muscle strength and imbalance using hand-held dynamometry in patients diagnosed with CMT and pes cavus, compared to healthy controls. METHODS: Muscles responsible for inversion, eversion, plantarflexion, and dorsiflexion of the foot and ankle were measured in 55 subjects (11 CMT patients with a frank pes cavus, and 44 healthy controls with normal feet) using the Nicholas hand-held dynamometer (HHD). Test-retest reliability of the HHD procedure also was determined for each of the four muscle groups in the healthy controls. RESULTS: Test-retest reliability of the HHD procedure was excellent (ICC3,1 = 0.88 to 0.95) and the measurement error was low (SEM = 0.3 to 0.7 kg). Patients with CMT were significantly weaker than normal for all foot and ankle muscle groups tested (p < 0.001). Strength ratios of inversion-to-eversion and plantarflexion-to-dorsiflexion were significantly higher in the patients with CMT and pes cavus compared to individuals with normal foot types (p > 0.01). CONCLUSIONS: Hand-held dynamometry is an objective and reliable instrument to measure muscle strength and imbalance in patients with CMT and a pes cavus foot deformity. PMID- 16045846 TI - Ankle arthrodesis with a retrograde femoral nail for Charcot ankle arthropathy. AB - BACKGROUND: Tibiocalcaneal arthrodesis with retrograde intramedullary nailing has gained acceptance as a salvage procedure for a multitude of ankle and hindfoot disorders and is frequently used in Charcot arthropathy of the ankle. Because of the severe osteopenia often associated with Charcot arthropathy of the ankle, an area of stress concentration leading to stress fracture at the proximal aspect of the nail has been identified. METHODS: To determine if this potential complication can be avoided, nine consecutive diabetic individuals with Charcot arthropathy of the ankle had ankle arthrodesis with a longer retrograde femoral nail. The average age of the patients was 52.3 years. Their average weight was 102.6 kg. RESULTS: Fusion was evident on radiographs in all nine patients at an average of 10.5 weeks. None of the patients developed a stress fracture or evidence of stress concentration at the proximal metaphyseal tip of the nails. One wound infection resolved after debridement and antibiotic therapy, and one postoperative hematoma resolved without surgery. At an average 32-month followup, all patients were ambulatory, using commercially available therapeutic footwear. None had developed a new foot ulcer, infection, or new episode of Charcot arthropathy. CONCLUSIONS: The use of a retrograde femoral nail for ankle arthrodesis in patients with Charcot arthropathy appears to decrease the risk of stress fracture compared with shorter nails without increasing the risk of other complications. PMID- 16045847 TI - Plantar forefoot pressure changes after second metatarsal neck osteotomy. AB - BACKGROUND: The aim of this study was to evaluate plantar pressure changes after second metatarsal neck osteotomy using the Weil technique. METHODS: Six below knee cadaver specimens were used. Each specimen was held in a custom-built apparatus and loaded to 500 N for a period of 3 seconds. Using a computerized Musgrave pedobarograph, pressure measurements were made before and after osteotomy in both neutral and 45-degree heel rise positions. All osteotomies were made at an angle of approximately 20 degrees relative to the long axis of the metatarsal shaft. The metatarsal heads were displaced proximally by 5 mm and fixed with a single Kirschner wire. RESULTS: After osteotomy there was an average decrease in pressure beneath the second metatarsal from 70.6 to 45.1 kPa in neutral and from 813.0 to 281.4 kPa in heel rise, representing statistically significant (p < or = 0.05) decreases of 36% and 65%, respectively. There also were significant decreases beneath the third metatarsal in both neutral (39%) and heel rise (37%), and beneath the fourth metatarsal in neutral position (28%). A significant pressure increase occurred beneath the first metatarsal in neutral (23%). No significant pressure changes occurred under the fifth metatarsal in either position. CONCLUSION: Overall, our results indicated that the Weil metatarsal neck osteotomy is effective at offloading the second metatarsal head at neutral and heel rise positions. PMID- 16045848 TI - Morton neuroma: comparative results of two conservative methods. AB - BACKGROUND: The initial treatment of Morton neuromas consists of conservative methods that include shoe modifications and steroid injections. The purpose of this prospective study was to compare the efficacy of these two methods to determine which is more effective as the initial treatment method. METHODS: Eighty-two patients with Morton neuromas were randomly assigned to receive either footwear modification with orthoses or steroid injections as initial treatment. Outcomes were evaluated at 1 month, 6 months, and 12 months. RESULTS: Patient satisfaction was significantly better (p < 0.01) in the group treated with steroid injections than those treated with shoe modifications at all three followup intervals. At 12-month followup, 82% of those treated with steroid injections had complete or partial relief of pain compared to 63% of those treated with footwear modifications alone. CONCLUSION: Steroid injections as initial treatment and shoe modifications with steroid injections at 6 months appear to give better results in Morton neuromas than shoe modifications alone, but the difference in the two groups were not statistically significant at one year followup. PMID- 16045849 TI - The anatomic features of the sural nerve with an emphasis on its clinical importance. AB - BACKGROUND: The sural nerve is formed by the union of the medial and lateral cutaneous nerves of the leg that originate from the tibial and common peroneal nerves. Operative procedures and traumatic injuries to the popliteal fossa, leg, ankle and foot place the sural nerve and its branches at risk. The aim of this study was to describe the course, variations and some clinically significant relations of the sural nerve. METHODS: The sural nerve was dissected in 30 lower limbs (leg-ankle-foot) of 15 cadavers. The specimens were measured, drawn and photographed. RESULTS: In 18 specimens (60%) the sural nerve originated from the union of the medial and lateral cutaneous nerves of the leg in the upper two thirds of the leg (classic type). The union of the medial and lateral cutaneous branches was in the distal third of the leg in three specimens (10%). The lateral cutaneous nerve was absent in five (16.7%), and the medial cutaneous nerve was absent in 2 (6.7%) specimens. In two specimens (6.7%) the nerves had separate courses. The mean distance between the most prominent part of the lateral malleolus and the sural nerve was 12.76 +/- 8.79 mm. The mean distance between the tip of the lateral malleolus and sural nerve was 13.15 +/- 6.88 mm. The most common distribution of the sural nerve in the foot was to the lateral side of the fifth toe (60%), followed by the lateral two and a half toes (26.7%). CONCLUSIONS: These described variations and measurements should be helpful for planning operative approaches that minimize the risk of sural nerve injury. PMID- 16045850 TI - Primary malignant non-Hodgkin lymphoma of the talus: a case report. PMID- 16045851 TI - Combined talar body, calcaneal, and navicular fracture: a case report. PMID- 16045852 TI - Aspergillus osteomyelitis and lymphangitis in immunocompromised patient after toenail clipping. PMID- 16045853 TI - Clinical tip: stabilization of the proximal Ludloff osteotomy. PMID- 16045854 TI - Immunodeficiencies: the clinician is not alone. PMID- 16045855 TI - Detection of four distinct groups of hen egg allergens binding IgE in the sera of children with egg allergy. AB - BACKGROUND: There appears to be a lack of agreement in the literature on the allergenicity of hen egg proteins. This may be partly due to the use of impure proteins in some cases. Egg yolk proteins have also been largely ignored in such studies. We therefore set out to determine, using especially purified proteins, their relative allergenicity, and to observe whether there were any relationships between their potency and the sensitivity of patients to them. METHODS AND RESULTS: The sera of 40 patients with clinically observed hen egg hypersensitivity were tested for specific IgE binding to purified egg white and egg yolk proteins using the radioallergosorbent test (RAST). Statistical treatment by correspondence analysis of the percent radioactive uptakes in the RAST to the 8 proteins demonstrated that there were four distinct groups of patients reacting in a similar way to four discrete sets of proteins. CONCLUSIONS: The first three sets of allergens consisted of egg white proteins as follows: firstly, lysozyme and ovalbumin; secondly, ovomucoid; and thirdly, ovomucin. The fourth set contained the egg white protein ovotransferrin and the egg yolk proteins apovitellenins I and VI and phosvitin. The existence of patient groups may explain why various workers have reported different allergens to be important in egg hypersensitivity. A sufficiently large number of patients must be examined so as to give a representative distribution across each group, otherwise the results may be biased towards one allergen. PMID- 16045856 TI - Aeroallergens sensitization in an allergic paediatric population of Cova da Beira, Portugal. AB - Cova da Beira is an interior central region of Portugal, with a population of 93,000 inhabitants. The first pollen counts performed in Portugal revealed the highest values of the country in this area. The aim of this study was to assess the aeroallergens sensitization in an allergic population, according to the age groups. In a 5 year period (1995-2000) 1790 consecutive outpatients were observed for suspected allergic symptoms. We included in this study all the 557 paediatric (< or = 15 years old) observed patients (317 male (57%) and 240 female (43%) with an average age of 7.6 < or = 4.2 years old). They were divided in three age groups (Group I: < or = 5 yr; Group II: 6-10 yr; Group III: 11-15 yr). 371 patients were submitted to skin prick tests to aeroallergens. 86.5% of these patients were sensitised to at least one allergen extract. The most representative aeroallergens sensitization were grasses mixture (44.9%), D. pteronyssinus (32.5%), D. farinae (29.1%), Olea europea (27.5%), Parietaria judaica (23.4%), cat dander (16.1%), Artemisia vulgaris (17.6%), Robinia pseudoacacia (12.2%), Platanus acerifolia (11.4%), Tilia cordata (11.4%) moulds mixture (11.2%), Plantago lanceolata (10.6%), dog dander (10.4%), and Pinus radiata (7.5%). The sensitisation to indoor aeroallergens, was similar in all age groups and it was less important than that of pollens. The prevalence of sensitisation to grasses was the greatest in all ages and the house dust mites sensitization was the second most prevalent. The highest pollens counts in this region could explain the early sensitisation even in young children. PMID- 16045857 TI - Different clinical and laboratory evolutions in ataxia-telangiectasia syndrome: report of four cases. AB - We report four patients with ataxia-telangiectasia syndrome that presented varied neurologic evolution. Three patients initially presented neurologic alterations of slow progression, evolving to late immunocompromised conditions. The fourth patient presented, from symptom onset, immune and neurologic debilitation, that were both severe and of fast progression. The chronological sequence of the most commonly observed immunocompromised conditions were in our patients, in ascending order, IgA deficiency, IgG2 deficiency and the neutrophil phagocytosis stage and common variable immunodeficiency. The first two reports are of sisters in whom the diagnosis was done between the ages of three and six years, having ocular apraxia, cerebellar ataxia and telangiectasia. Slow progression of neurologic debilitation was observed, without presentation of intermittent infections. The patients began presenting accentuated immunocompromised conditions at the ages of 14 and 17 years, dying at the ages of 16 and 20 years, respectively, due to severe infections that were resistant to treatment. The diagnosis of the third case was established when the patient was two years old, presenting ataxia and telangiectasia. Syndrome progression was slow, presenting at the age of eight years more accentuated neurologic disorders and IgA deficiency. The fourth case presented significant neurologic compromise at the age of five, simultaneous to IgA and IgG2 deficiency, and repeating pneumonias and sinusitis. At this time, intravenous gammaglobulin reposition was done. The neurologic and immune disorders progressed rapidly, and at the age of eight presented the inability to walk. At this time inversion of the CD4/CD8 ration was verified through laboratory tests. PMID- 16045858 TI - The effects of long term use of inhaled corticosteroids on linear growth, adrenal function and bone mineral density in children. AB - In this study we aimed to investigate the long term effects of inhaled steroids on linear growth, adrenal function and bone mineral density. Thirty children with moderate asthma were randomly divided into two groups. Fifteen children (8 boys, 7 girls mean age; 10.6 +/- 2.1) were treated with budesonide (group 1), and 15 (9 boys, 6 girls, mean age; 9.6 +/- 2.4). with fluticasone propionate (group 2). Control group included 30 children. Anthropometric assessment, symptom and medication scores, pulmonary functions, bone mineral density, serum and urine cortisol levels and ACTH stimulation test were evaluated at the beginning of the study and after one year period. Symptom and medication scores, pulmonary functions improved significantly in both groups (p < 0.05). The mean annual growth was similar in group 1 and 2 and control group. Bone mineral density was comparable with control group at the beginning of the study and after one year. Mean serum cortisol level diminished at the end of the therapy but no significant differences were found between the initial and end values in respect to urine cortisol levels and cortisol/creatinine ratio. Of three groups ACTH stimulation test revealed that there were no significant difference between study and control groups. In conclusion, although we did not observed any side effects of inhaled corticosteroids we suggest that children treated with inhaled corticosteroids for a long time should be followed closely with respect to side effects. PMID- 16045859 TI - Frequency in allergy to proteins of latex in health care workers. AB - OBJECTIVE: The objective of this study is to evaluate allergy prevalence to latex proteins in health care workers at the Laboratory and Surgery Room of Hospital CIMA Chihuahua. METHODOLOGY AND RESULTS: A thorough clinical chart was recorded for all health care workers studied: hematic biometry, total IgE by ELISA method, specific IgE to latex antigen by ELISA (pharmacy CAP system), cutaneous tests with latex antigen (Aphi de Mexico, Hevea Brasiliensis Biopal Inc. Spoken WA), through scarification, together with histamine and Evans (positive-negative control). The number included is 99 individuals. Specific IgE to latex in 4 cases was positive (4%) and in the cutaneous tests to latex, 24 cases (24%) resulted positive. CONCLUSIONS: The study reports a prevalence of (4%) when performing the specific IgE to latex and (24%) to the cutaneous test with antigen to total latex. This data allows us to continue evaluating the personnel at risk at the hospital, with better results in the administration of Labor Medicine at this medical institution. PMID- 16045860 TI - [Exposition and sensitization to Tyrophagus putrescentiae in a allergic population to Dermatophagoides pteronyssinus in Huelva, Spain]. AB - BACKGROUND: In this work we analyzed the allergological importance of Tyrophagus putrescentiae (Tp) in Huelva (SE Spain). We studied the level of exposition and the grade of sensitization to Tp in a group of patients sensitized to Dermatophagoides pteronyssinus (Dpt). The allergenic cross-reactivity between Dpt and Tp was determined by RAST inhibition. METHODS AND RESULTS: We analyzed house dust samples from the dwellings of allergic patients with documented Dpt sensitization. Skin test (ST), conjunctival provocation (CP) and/or specific IgE (RAST) to Tp were performed when Tp was identificated in the house dust sample of the patient. Among the 136 dust samples studied, Dpt was the most frequently identified mite species (94.8%) and Tp was found in third position (41.1%) after Glycyphagus domesticus (54.4%). Among the 45 patients studied, 23 (51.1%) presented, at least, two positive tests, 18 (40%) were not sensitized to Tp and 4 (8.8%) showed contradictory results. 26 patients (57.7%) inhabited in urban areas and 19 (42.2%) in rural regions. We determined specific IgE (RAST) to Tp in 25 patients, and the results were positive in 12, with only 7 with values greater than 2 kU/L. No significant correlation were found between IgE-antibody levels to Dpt and Tp. The RAST inhibition studies confirmed the low cross-reactivity between these mites and only in one patient Dpt partially inhibited the IgE binding to Dpt extract. CONCLUSIONS: Tp was the second more frequent storage mite in the house dust samples from patients allergic to Dpt in Huelva. However, only half of the exposed patients were sensitized to Tp and the majority inhabited in urban areas. No significant correlation were found between IgE-antibody levels to Dpt and Tp. The RAST inhibition studies confirmed the low cross-reactivity between these mites. PMID- 16045861 TI - Safety and tolerability of ultra-rush (20 minutes) sublingual immunotherapy in patients with allergic rhinitis and/or asthma. AB - BACKGROUND: The safety and good tolerability of sublingual immunotherapy (SLIT) has already been proved in allergic patients, but only one study has investigated the occurrence of immediate adverse reactions in allergic patients after a 2-hour ultra-rush regimen of SLIT performed with a chemically modified extract (sublingual monomeric allergoid, Lais, Lofarma S.p.A., Milan). The objective of the present study was to evaluate the occurrence of immediate adverse reactions in allergic patients after a very fast (20 minutes) ultra-rush regimen of sublingual allergoid SLIT. METHODS AND RESULTS: We studied 105 patients: 28 children (20 male, mean age 13.3 +/- 2.1 yr) and 77 adults (29 male, mean age 34.7 +/- 9.9 years) with a history of intermittent/persistent rhinitis or intermittent/mild persistent asthma due to House Dust Mite (n = 56), Parietaria (n = 34) and Timothy-grass (n = 15) The build-up ultra-rush phase involved the administration, every five minutes, of increasing doses of the sublingual allergoid SLIT. All patients tolerated the treatment very well. Only one patient out of 105 (0.9%) had a mild local symptoms (gastric pirosis) that occurred 30 minutes after the last initial dose and spontaneously disappeared as the treatment was continued. CONCLUSIONS: These data show the excellent safety and tolerability profile of an ultra-rush SLIT regimen performed with a chemically modified extract, even when high doses were administered through an extremely short induction phase (20 minutes), thus confirming the previously reported results. PMID- 16045862 TI - [Safety of rush immunotherapy with Hymenoptera venom]. AB - OBJECTIVES: To determine the incidence and nature of adverse events associated with the induction of rush Hymenoptera venom immunotherapy. MATERIAL AND METHODS: Between 1998 and 2003, we administered venom immunotherapy to 48 patients allergic to bee or wasp venom, by means of a rush immunotherapy protocol (3 days). RESULTS: We observed no severe adverse reactions in any patients. 12 patients developed only local reactions at the site of injections that did not required any pharmacological treatment. Two patients experienced mild systemic reactions consisting of diffuse urticaria on day 3. Both adverse reactions were treated with intravenous antihistamines. CONCLUSIONS: Our experience confirms that rapid venom immunotherapy is safe and should be considered in every case especially for patients during the stinging insect season when a rapid protection is required. PMID- 16045863 TI - Angioedema induced by inhalation of vapours from cooked white bean in a child. AB - BACKGROUND: There are few references of allergic reaction to beans in childhood. We report the case of a seven years old boy who suffered from angioedema associated to inhalation of vapours from cooked white bean. METHODS: skin prick tests (SPT) were performed by prick-by-prick with cooked white bean and legumes. It was also determined total IgE and specific IgE antibodies to bean and legumes with the use of the CAP enzymo-immunoassay. Subsequently, a oral challenge test was carried out with white bean. RESULTS: The prick-by-prick with white bean was positive in our case, and negative in ten controls patients. Specific IgE in patient serum, assayed by CAP was positive for white bean and green bean. The patient developed angioedema after ingestion cooked white bean. CONCLUSION: we demonstrated a type I hypersensitivity to white bean in a seven years old child by SPT, specific IgE antibodies and challenge test. PMID- 16045864 TI - [Treatment with nebulized lidocaine in steroid-dependent asthma]. AB - BACKGROUND: The management of glucocorticoid dependent and resistant asthma encompasses a group of problems, affecting as much control of clinical symptoms as the onset of adverse effects to prolonged use of steroids. MATERIALS AND METHODS: Woman 52 years old, steroid dependent asthma fifteen years ago. She has important adverse effects in consequence of the use of systemic glucocorticoids. The employment of methotrexate attained decrease in the dosage of steroids temporarily. The worsening of the clinical situation and deterioration of secondary effects by use of steroids necessitated treatment with nebulized lidocaine. A 2% solution to initial dosage was administered three times daily. The maximal dosage needed was 80 mg three times daily. RESULTS: After three months of lidocaine treatment, the patient was able to reduce oral steroids eventually discontinuing these drugs, and the effects secondary to exogenous hypercortisolism disappeared. Moreover, an improvement in clinical symptoms and quality of life was observed with stability in the pulmonary function. CONCLUSIONS: The use of nebulized lidocaine could be useful and provide alternatives in patients with severe asthma. PMID- 16045865 TI - An unusual adverse drug reaction? AB - INTRODUCTION: Adult Still Disease (ASD) is a rare (incidence 1-1,6/1,000,000 in Europe) seronegative polyarthropathy diagnosed with clinical criteria, excluding other etiologies. Minocycline, a semisynthetic derivative of tetracycline, has been associated with many adverse effects. We present the case of a 18-years-old man with a high suspicion of an adverse drug reaction (ADR), that was finally diagnosed of ASD. MATERIAL AND METHODS: In order to exclude other diseases, laboratory test, radiology and allergological studies were performed. The results of the allergological studies excluded the possibility of an ADR. The rest of the results determined the diagnosis of ASD. CONCLUSION: It's necessary to emphasize the importance of an accurate differential diagnosis in cases like this, because many diseases may mimic an ADR, and may be underdiagnosed (or misdiagnosed). PMID- 16045866 TI - Cutaneous reactions due to diltiazem and cross reactivity with other calcium channel blockers. AB - BACKGROUND: The spectrum of cutaneous eruptions in association with calcium channel blockers is extensive, varying from exanthemas to severe adverse events. Reactions due to diltiazem occur more frequently than with other calcium channel blockers. Patch testing has been used as confirmatory testing in patients with extensive cutaneous reactions. Cross-reactivity among these drugs have not been established. MATERIAL: We present 3 patients: 1) A 54-year-old man developed a generalized erythema-multiforme-like reaction followed by erythrodermia and exfoliative dermatitis 6-7 days after starting on diltiazem. The drug was stopped and remission was obtained with emollients and systemic corticosteroids and antihistamines within 12 days. 2) A 80-year-old woman experienced a pruritic exanthematous eruption on her trunk which evolved to generalized erythrodermia and superficial desquamation. This reaction appeared 10 days after taking diltiazem, and gradually improved in 10-12 days after discontinuation of this drug. 3) A 79-year-old man presented with erythema and pruritus initially on the back, and then affecting thorax, extremities and face. He had started treatment with diltiazem three days before. Diltiazem was stopped and steroid and antihistamine therapy was given. His skin condition improved, but 3 days later the patient received verapamil with worsening of previous situation. He recovered within 7 days. METHODS AND RESULTS: Two to six months after the reaction, we carried out epicutaneous tests with calcium channel blockers from different groups. Diltiazem proved positive (at 48 and 96 hours) in the three patients; nifedipine was also positive in patient 2, and verapamil in patient 3. Controlled administration of verapamil was well tolerated in patient 2 after the reaction, and the patient 1 has taken nifedipine without problems. CONCLUSIONS: 1) We report 3 cases of cutaneous reactions due to diltiazem. 2) Epicutaneous tests have been useful for diagnosis. 3) As one of patients had positive patch tests to diltiazem and nifedipine, and other one with diltiazem and verapamil, more studies are needed to demonstrate cross reactions among calcium channel blockers. PMID- 16045867 TI - [Pediatric environmental health in Spain. Where are the pediatricians?]. PMID- 16045868 TI - [Risk factors for mortality in very low birth weight infants with respiratory distress syndrome]. AB - INTRODUCTION: In the last few years the life expectancy of very low birth weight (VLBW) infants has improved. When these patients have respiratory distress syndrome, it is difficult to know with any certainty which factors have the greatest influence on mortality. The aim of this study was to determine which variables, among a series, have the greatest influence on mortality from a multivariate perspective. PATIENTS AND METHODS: A sample of 209 VLBW infants born over a long period (15 years and 7 months) was studied. The following variables were analyzed: date of birth, degree of respiratory distress syndrome, sex, birth weight, weeks of gestation, born within or elsewhere, prenatal corticoid administration, type of gestation, type of delivery, amniorrhexis time, Apgar test at 1 and 5 minutes, surfactant administration, hours of life at which the first dose of surfactant was administered, and early sepsis. A multiple logistic regression analysis was developed using Hosmer-Lemeshow methodology. RESULTS: The following variables were identified as significant: birth weight, Apgar test at 5 minutes, prenatal corticoids, degree of respiratory distress syndrome, and surfactant administration. The remaining variables were less important in the multivariate analysis. CONCLUSIONS: Higher birth weight and Apgar score at 5 minutes, prenatal corticoid and surfactant administration, and a lower degree of respiratory distress syndrome reduce mortality. The logistic regression model used quantifies how these factors behave and allows the probability of mortality in VLBW infants with respiratory distress syndrome to be estimated. PMID- 16045869 TI - [Birth weight and familial smoking]. AB - INTRODUCTION: Tobacco smoking during pregnancy is an important risk factor in the pediatric population and has been associated with an increase in low birth weight (LBW) infants. PATIENTS AND METHODS: We carried out a retrospective case-control study of infants admitted to the Infants Department of the Hospital Universitario Infantil Nino Jesus in Madrid. Data from 2370 infants admitted to the hospital between 2002 and 2004 were collected. RESULTS: The odds ratio (OR) for having a LBW infant was 1.42 (95 % CI: 1.017-1.985) among mothers who smoked during pregnancy and was 1.37 (95 % CI: 1.014-1.863) among women whose partners smoked. Gestational age was also a predictive factor of birth weight: the OR was 0.585 (95 % CI: 0.545-0.628). No significant differences for risk of LBW were found between sexes (OR: 1.25; 95 % CI: 0.934-1.671). CONCLUSIONS: Our results should reinforce the importance of smoking prevention during and after pregnancy in both parents, which could reduce many complications in children's health with a high medical, social and economic cost. PMID- 16045870 TI - [Hospitalizations for varicella in the Hospital Infantil La Fe, Valencia,Spain, 2001-2004]. AB - INTRODUCTION: Varicella is a common, highly contagious disease. It is usually benign but has potentially serious complications. PATIENTS AND METHODS: To assess the clinical characteristics and the associated cost of varicella hospitalization, we reviewed the medical records of children hospitalized for varicella between 2001 and 2004. Children with coincidental varicella hospitalized for a different reason were excluded. RESULTS: Of 1177 children with varicella attended at the emergency room, 101 (8.6 %) were hospitalized. The median age was 3.2 years (21 days to 18.9 years). Twenty-eight children had underlying disease. Thirty-seven children had no complications and the reason for admission was: a) risk of severe varicella (21 immunocompromised children, three neonates), and b) high fever or observation (13 cases). The 64 remaining children were admitted for 66 complications of varicella. The most common complications were skin/soft tissue infections (33 patients) and the leading cause was Streptococcus pyogenes (n = 13) and Staphylococcus aureus (n = 10) isolated in blood or the site of infection. Other complications were pneumonia (13 children), neurological (febrile seizures in nine, meningoencephalitis in two, acute disseminated encephalomyelitis in one, cerebellitis in one), hematological (neutropenia in one, Henoch-Schonlein purpura in one and thrombopenic purpura in three) and osteoarticular (synovitis in one and septic arthritis in one). One patient died of multiorgan failure. During the study period, the rate of emergency room visits due to varicella doubled and the number of admissions for complications tripled. The mean length of hospital stay was 6.8 days (range: 1-28 days) and the total associated cost was 397,314.14 Euro, excluding symptomatic treatment. CONCLUSIONS: The high morbidity associated with varicella and its complications, as well as the high social costs of this disease, support the implementation of routine varicella vaccination. This could reduce the total number of cases, their severity, direct costs, generated by medical care, and indirect costs, generated by the disease and hospitalization. PMID- 16045871 TI - [Epidemiology of community-acquired pneumonia in children aged less than 5 years old in the Autonomous Community of Valencia (Spain)]. AB - OBJECTIVE: To assess the incidence of community acquired pneumonia in the Autonomous Community of Valencia in Spain, and describe its treatment and complications. METHODS: A retrospective cohort comprising 654 children born in 1995 and 1996 in Valencia and followed-up during the first 5 years of life by nine pediatricians was studied. The number of cases of pneumonia, treatment, complications and interventions was recorded. RESULTS: Ninety-nine episodes of community-acquired pneumonia in 80 children were recorded (1.24 cases/child). Fifty-one cases (51.1 %) occurred before the child's third birthday and 38 (38.4 %) occurred between the third and the fourth. Of the 99 cases, 51 were diagnosed in primary care and 46 in the emergency room. There was a mean of 2.44 visits per process in primary care (range 1-6). All the children were treated with antibiotics. The most frequently used were amoxicillin-clavulanate (43.3 %) and cefuroxime (26.3 %). Fourteen patients required a change of antibiotic. Twenty three percent of the children were hospitalized. CONCLUSION: The incidence of community-acquired pneumonia in Valencia was 30.3 cases/1000 children aged less than 5 years/ year (95 % CI: 18.7-46.8), and the incidence of hospitalization was 7.03 cases/1000 children aged less than 5 years/year. PMID- 16045872 TI - [Clinical and epidemiological study of community-acquired pneumonia in children aged less than 5 years old]. AB - OBJECTIVE: To describe the clinical characteristics of community-acquired pneumonia and associated morbidity in children under 5 years old. PATIENTS AND METHOD: We performed a prospective epidemiological study in 12 primary care clinics and two pediatric emergency departments in Vizcaya, Spain. Demographic, clinical, radiological, laboratory and treatment data were recorded at diagnosis. Different pneumonia groups were established on the basis of radiological images (lobar pattern), total leucocyte count (> 15,000/ml) and C-reactive protein value (> 80 mg/l). These groups ranged from definitive pneumococcal pneumonia (pneumococcus isolated in usually sterile specimens) to pneumonia of probable non pneumococcal etiology. All patients were followed-up for 7 to 15 days after diagnosis to ascertain outcome. RESULTS: Between February and April 2003, 412 children with pneumonia were enrolled. The mean age was 33.4 6 15.34 months and 21 % of the patients had received the pneumococcal conjugate vaccine. Overall 20 % had general malaise and 14.6 % required admission, with a mean length of hospital stay of 4.10 +/- 2.21 days. The mean length of treatment was 9.51 +/- 2.44 days and the mean number of visits to the primary care pediatrician was 2.02 6 1.10. Fifty-three patients (12.9 %) had definitive or highly probable pneumococcal pneumonia and these children had higher temperature (38.74 +/- 0.84 vs 38.38 +/- 0.94 degrees C), a higher percentage of general malaise (50.9 % vs 15.3 %) and a higher hospitalization rate (41.5 % vs 10.5 %). CONCLUSIONS: Community-acquired pneumonia in children under 5 years old provokes high morbidity. According to clinical, radiological and laboratory data, suspected pneumococcal pneumonia seems more severe than forms probably caused by other agents. PMID- 16045873 TI - [Contribution of flexible bronchoscopy to the diagnosis of upper airway alterations]. AB - INTRODUCTION: Flexible bronchoscopy (FB) is becoming an increasingly common diagnostic and therapeutic technique and is currently an essential procedure in pediatric pulmonology departments. Because the procedure is easy to perform under sedation and topical anesthesia, avoiding surgery with general anesthesia, which is required for rigid bronchoscopy, it has allowed the use of airway endoscopy in infants and children to be increased and has reduced patient risks. OBJECTIVE: To analyze the contribution of FB to the diagnosis of upper airway alterations and to characterize the frequency and type of upper airway anomalies. PATIENTS AND METHODS: We retrospectively reviewed the FB performed in our center between January 1993 and March 2003 in children aged less than 14 years old. RESULTS: A total of 456 FB were performed in 378 children. One hundred eleven FB (24.5 % of all endoscopies) were required for suspected upper airway anomalies and 55.8 % corresponded to stridor. Upper airway inspection revealed abnormality in 30.7 % (140 FB). There were no severe complications. CONCLUSION: FB is useful in the diagnosis of upper airway abnormalities. The main indication for FB is stridor and the most common abnormal finding of the upper airway is laryngomalacia. PMID- 16045874 TI - [Pediatric environmental health specialty units in Europe: from theory to practice]. AB - BACKGROUND: The World Health Organization encourages the development of strategies to identify, increase awareness and solve problems of Pediatric Environmental Health (PEH) in units and centers of excellence. Pediatricians are in an excellent position to identify children at environmental risk, advise parents about the best way to reduce or prevent these risks, and recommend changes in health policies to politicians. OBJECTIVE: To inform pediatricians about the underlying principles and activities of pediatric environmental health specialty units (PEHSU) in Spain and Europe. To analyze up-to-date knowledge and the steps required for the development of PEH in Spain and Europe. MATERIAL AND METHODS: Review of the strategies, initiatives and examples of useful practices for national and international organizations involved in the development and implementation of PEH. Report of activities of PEHSUs. RESULTS: In Spain and Europe pediatricians do not receive training in PEH. PEHSUs are centers specialized in the diagnosis, treatment, and prevention of environmentally related diseases. These centers are composed of multidisciplinary teams of health and other professionals, coordinated by pediatricians specialized in PEH, that have the study of environment-related pediatric problems as a common denominator. The activity of these units emphasizes teaching, research, scientific activity, the development of environmental history in pediatrics, diagnosis and treatment of the "pediatric environmental wound", assessment of risks in pediatrics, and specialization in the critical review of the literature on PEH to train pediatricians to be experts in environmental health. The activities and fields of PEHSU can be summarized as: medical care, training, research, and community health (school health). CONCLUSIONS: PEH should be considered by Ministries of Health and Pediatrics Associations as an important emergent area for current and future generations of pediatricians. Thus, it is necessary: a) to develop an integral plan for pediatric training in PEH; b) to actively seek funding and support to increase the presence of PEHSUs in Europe, and c) to create expert committees in PEH in National Associations of Pediatrics in Europe. PMID- 16045875 TI - [End-of-life care in the pediatric intensive care unit. A literature review]. AB - INTRODUCTION: In the last few years, there has been growing concern in the literature about issues related to end-of-life care in pediatric intensive care units (PICUs), with special attention on the family/patient unit, communication, and a dignified death. OBJECTIVE: To evaluate the experience and development of end-of-life care in PICUs through a literature review, by determining the type of studies that have been performed, their topics, the issues discussed, and their development in the last few years. MATERIAL AND METHODS: Review of the medical literature in Medline and the database of the National Library of Medicine Gateway, using the key words from MeSH: "end of life", "pediatric intensive care", "critical care", "palliative care", "death", and "compassionate care". The earliest year of the search was 1990. The languages selected were English and Spanish. Inclusion criteria were the relationship with the topic to be studied, excluding articles with no abstract. Additional searches were made of references in selected articles. RESULTS: Eighty-one articles were retrieved from the initial search. Of these, 43 were selected as the most relevant investigations in end-of-life care in ICUs and 18 placed special emphasis on the PICU. More than half of the articles (62 %) were reviews and the remaining articles were descriptive or observational studies. The number of publications increased after 1995. Most of the studies were performed in the USA or Canada and only three studies were performed in Spain. CONCLUSIONS: In the last few years, several studies have been performed that reveal increasing concern about limits to therapeutic intervention and the need to improve end-of-life care in the PICU setting. PMID- 16045876 TI - [Percutaneous tracheostomy under endoscopic guidance in adolescents]. AB - Percutaneous tracheostomy (PT) is a well-established procedure in adult patients who require long-term airway control. Because experience with this procedure is scarce in pediatrics, we report the results of PT by means of a progressive dilatational method with endoscopic guidance in three critically-ill adolescents with severe head injury. PT was performed by means of the Ciaglia Blue Rhino method at the bedside, guided by fiberoptic bronchoscopy by the pediatric intensive care unit staff. No hemodynamic or respiratory compromise was observed during the procedure. No complications were attributable to PT. Two patients were decannulated without incidents. The reported cases indicate that PT can be performed safely, at least in adolescents. PMID- 16045877 TI - [Bacterial tracheitis: an infectious cause of upper airway obstruction to be considered in children]. AB - INTRODUCTION: Bacterial tracheitis is a cause of severe upper respiratory airway obstruction. It has been considered a rare entity whose clinical and epidemiological features are not well known. PATIENTS AND METHODS: The medical records of children diagnosed with bacterial tracheitis and admitted to our pediatric intensive care unit (PICU) from June 1992 to May 2004 (12 years) were analyzed. The following variables were recorded: age, sex, personal history, need for endotracheal intubation, duration of intubation, isolated bacteria, antibiotic treatment, steroid administration, and length of stay in the PICU. RESULTS: Twelve patients (six girls and six boys) were included in this study. Age ranged from 1 month to 13 years (median 24 months). Four children had a history of acute laryngotracheitis, one had Down syndrome and another had interauricular communication. All patients needed orotracheal intubation for 2 to 72 hours (median 48 hours). All patients received empirical antibiotic therapy: cephalosporins in 11 children (91 %) (in association with vancomycin or teicoplanin in four of them) and erythromycin in one (9 %). Corticosteroids were administered in nine patients (75 %). Bacterial pathogens were isolated from tracheal secretion culture in seven children (58 %) and the most common was Staphylococcus aureus (42 %). Three children (25 %) developed lobar pneumonia and one developed congestive cardiac insufficiency as a complication of bacterial tracheitis. In all patients, clinical outcome was satisfactory. CONCLUSIONS: Although the absolute frequency of bacterial tracheitis is low, in our environment this entity represents a considerable cause of severe upper respiratory airway obstruction secondary to an infectious process. Therefore, it should be considered in patients who present with stridor and fever. If the correct treatment is provided, the prognosis is generally good. PMID- 16045878 TI - [Chronic maxillary atelectasis: an uncommon cause of persistent radiological opacification]. AB - Chronic maxillary atelectasis (CMA) is an infrequent entity characterized by a persistent decrease in maxillary sinus volume due to inward bowing of one or more antral walls. The cause of CMA is probably maxillary sinus hypoventilation secondary to ostiomeatal complex obstruction. This creates negative pressure, leading to thinning of the sinus walls. We report the case of a 14-year-old asymptomatic girl with an opacified maxillary sinus on radiology following unsuccessful antibiotic treatment. A coronal computed tomography scan showed partial right antral opacification, with a laterally malpositioned medial infundibular wall and fontanel retraction, and consequently CMA was diagnosed. The interest of this case lies in the infrequency of CMA and the need to distinguish it from maxillary sinusitis. PMID- 16045879 TI - [Benign intracranial hypertension and heterozygosity for factor V Leiden mutation]. AB - Benign idiopathic intracranial hypertension (BIH) in association with prothrombotic conditions has been reported with increasing frequency in the medical literature. Recently, activated protein C resistance (APCR) has been identified as a factor in some cases. Because of its high prevalence, factor V Leiden mutation (FVL) is the most frequent coagulation abnormality associated with cerebral venous thrombosis. Reduced craniospinal fluid reabsorption due to damaged arachnoid villi secondary to microthrombus formation has been proposed as an explanation for the physiopathology of BIH and FVL. We describe two patients with a diagnosis of BIH, in whom the only risk factor was heterozygosity for FVL mutation. PMID- 16045880 TI - [Inhaled iloprost: a therapeutic alternative for persistent pulmonary hypertension of the newborn]. PMID- 16045881 TI - [Multiple pulmonary abscesses due to Staphylococcus aureus in a patient with a Port-A-Cath]. PMID- 16045882 TI - [Epidural empyema after laparoscopy for hypotrophic pyloric stenosis]. PMID- 16045883 TI - [Probable methemoglobinemia following EMLA administration]. PMID- 16045884 TI - [Cyclic neutropenia with anti-NA2 antibodies and treatment with recombinant granulocyte colony-stimulating factor]. PMID- 16045885 TI - [Congenital triangular alopecia]. PMID- 16045886 TI - [Early gender assignment in cases of genital ambiguity at birth]. PMID- 16045887 TI - [Popliteal mass associated with osteochondroma]. PMID- 16045888 TI - Growth hormone receptor gene expression in the skeletal muscle of normal and double-muscled bovines during foetal development. AB - The expression of the growth hormone receptor (GHR) gene was investigated in semitendinosus muscle during bovine foetal development in both normal and double muscled Charolais foetuses which differ with respect to muscle development. Northern-blot analysis of foetal muscle RNA preparations with a GHR cDNA probe identified the 4.5 kb GHR mRNA as early as 130 days post-conception. In double muscled animals, the expression of GHR mRNA increased from 130 to 210 days of gestation while it stayed stable in normal ones. It was significantly higher (P < 0.05) in double-muscled foetuses compared to normal ones from the second third of gestation. Northern-blot analysis of foetal muscle RNA preparations from both genotypes with a beta-actin cDNA probe, revealed lower beta-actin gene expression in double-muscled foetuses than in normal ones, suggesting a delay in the differentiation of muscle cells. In situ hybridisation revealed the localisation of specific GHR mRNA in muscle cells at all gestation stages analysed (130, 170, 210 days post-conception) but not in connective tissue surrounding the muscle cells. At the adult stage, the hybridisation signal was also very high and observed in muscle cells only. These results show the ontogeny of GHR mRNA in bovine muscle and demonstrate a difference between normal and double-muscled animals. PMID- 16045889 TI - Dose effect of alpha-linolenic acid on lipid metabolism in the hamster. AB - In order to meet dietary requirements, the consumption of alpha-linolenic acid (ALA, 18:3 n-3) must be promoted. However, its effects on triglyceride (TG) and cholesterol metabolism are still controversial, and may be dose-dependent. The effects of increasing dietary ALA intakes (1%, 10%, 20% and 40% of total FA) were investigated in male hamsters. ALA replaced oleic acid while linoleic and saturated FA were kept constant. Triglyceridemia decreased by 45% in response to 10% dietary ALA and was not affected by higher intakes. It was associated with lower hepatic total activities of acetyl-CoA-carboxylase (up to -29%) and malic enzyme (up to -42%), which were negatively correlated to ALA intake (r(2) = 0.33 and r(2) = 0.38, respectively). Adipose tissue lipogenesis was 2-6 fold lower than in the liver and was not affected by dietary treatment. Substitution of 10% ALA for oleic acid increased cholesterolemia by 15% but, as in TG, higher ALA intakes did not amplify the response. The highest ALA intake (40%) dramatically modified the hepatobiliary metabolism of sterols: cholesterol content fell by 45% in the liver and increased by 28% in the faeces. Besides, faecal bile acids decreased by 61%, and contained more hydrophobic and less secondary bile acids. Thus, replacing 10% oleic acid by ALA is sufficient to exert a beneficial hypotriglyceridemic effect, which may be counteracted by the slight increase in cholesterolemia. Higher intakes did not modify these parameters, but a very high dose resulted in adverse effects on sterol metabolism. PMID- 16045890 TI - Testing the antagonistic effect of follistatin on BMP family members in ovine granulosa cells. AB - Follistatin was first demonstrated as an activin-binding protein, neutralizing its actions. However, there is emerging evidence that follistatin inhibits the action of other members of the transforming growth factor beta(TGFbeta) / bone morphogenetic protein (BMP) superfamily. Recently, numerous BMP factors have been shown to play important roles in regulating folliculogenesis and ovulation rate in mammals, and such a potential antagonistic role of follistatin is of particular interest in the context of ovarian function. Using a biological test based on progesterone production by ovine primary granulosa cells in culture, we show that follistatin was a strong antagonist of activin A, but not BMP-2 or BMP 4 actions. In contrast, noggin, a known specific BMP antagonist, had no effect on activin A but strongly neutralized BMP-2 and BMP-4 actions. BMP-6 action was only slightly reduced by both follistatin and noggin. Our data led to the conclusion that follistatin would not represent a determinant physiological modulator of the biological effect of BMP factors on granulosa cells. PMID- 16045891 TI - Effect of feeding live yeast products to calves with failure of passive transfer on performance and patterns of antibiotic resistance in fecal Escherichia coli. AB - Fifty-two newborn Holstein calves with serum IgG concentrations less than 0.73 g.dL(-1) were randomly assigned to one of four treatments: no added live yeast (control), 0.5 g of live yeast added to the grain for 84 d (SC; Saccharomyces cerevisiae), 0.5 g of live yeast added to the milk for 42 d (SB; S. cerevisiae, spp. boulardii), and 0.5 g of live yeast added to the grain for 84 d and to the milk for 42 d (SCSB). Calves were offered 440 g of milk replacer DM for the first 42 d and grain for ad libitum intake throughout the study. Plasma was analyzed weekly for concentrations of glucose and beta-hydroxybutyrate. Escherichia coli isolated from fecal samples collected every 2 weeks were used for determination of antibiotic resistance patterns. Calves receiving SC consumed more grain DM, had increased weight gain prior to weaning, and increased plasma glucose concentrations compared to controls. Days with diarrhea were reduced by feeding live yeast to calves. Antibiotic resistance in fecal E. coli was associated with the age of calves with highest levels of resistance observed in the 3 d calves. While calves receiving SCSB had higher levels of antibiotic resistance than controls, this effect was not associated with any of the other treatments. Improvements in performance of calves with failure of passive transfer were observed when live yeast was added only to the grain. PMID- 16045892 TI - In vitro metabolism of rumenic acid in bovine liver slices. AB - Ruminant products are the major source of CLA for humans. However, during periods of fat mobilization, the liver might play an important role in CLA metabolism which would limit the availability of the latter for muscles and milk. In this context, rumenic acid (cis-9, trans-11 CLA) metabolism in the bovine liver (n = 5) was compared to that of oleic acid (n = 3) by using the in vitro liver slice method. Liver slices were incubated for 17 h in a medium containing 0.75 mM of FA mixture and 55 microM of either [1-(14)C] rumenic acid or [1-(14)C] oleic acid at 37 degrees C under an atmosphere of 95% O(2)-5% CO(2). Rumenic acid uptake by liver slices was twice (P = 0.009) that of oleic acid. Hepatic oxidation of both FA (> 50% of incorporated FA) led essentially to the production of acid-soluble products and to a lower extent to CO(2) production. Rumenic acid was partly converted (> 12% of incorporated rumenic acid) into conjugated C18:3. CLA and its conjugated derivatives were mainly esterified into polar lipids (71.7%), whereas oleic acid was preferentially esterified into neutral lipids (59.8%). Rumenic acid secretion as part of VLDL particles was very low and was one-fourth lower than that of oleic acid. In conclusion, rumenic acid was highly metabolized by bovine hepatocytes, especially by the oxidation pathway and by its conversion into conjugated C18:3 for which the biological properties need to be elucidated. PMID- 16045893 TI - Nutritional value of meat: the influence of nutrition and physical activity on vitamin B12 concentrations in ruminant tissues. AB - An important nutritional characteristic of ruminant meat is its high content in vitamin B12. The variability of these contents is not known. Three studies were been set up in order to test the influence of the animal species (2 studies on Charolais steers slaughtered at 30-32 months of age, n = 24 and n = 30 and a third one on lambs slaughtered at 4.5 months of age, n = 21), of the nature of the diet (grass vs. maize silage, lucerne or concentrate diets) and of physical activity (without or with walking) on the vitamin B12 contents of different muscle types (rather oxidative (Rectus Abdominis, RA), intermediate (Longissimus Dorsi, LD), or glycolytic (Semi Tendinosus, ST)) and on the liver. The animals were supplemented in macro and trace minerals according to usual feeding practices in France in order to theoretically avoid any risk of deficiency. For this reason, cobalt allowances, which are necessary for the ruminal synthesis of vitamin B12, could differ among treatments. The results indicate the following: (1) cobalt allowances varied widely among treatments, from (sub-)deficient to plethoric allowances, influencing vitamin B12 contents of the liver, and muscles (only in case of deficiency), (2) the effects of dietary treatments or of physical exercise were essentially related to differences in cobalt allowances, (3) the oxidative type muscle (RA) showed contents which were double those in glycolytic type muscle (RA 10.8 vs. ST 5.0 ng.g(-1)) and (4) vitamin B12 contents of raw muscles were lower than the values indicated in tables of feed composition for humans for cooked meat (0.5 to 1 vs. 2 to 3 microg.100 g(-1)). PMID- 16045895 TI - The minerals of milk. AB - The salt of milk constitutes a small part of milk (8-9 g.L(-1)); this fraction contains calcium, magnesium, sodium and potassium for the main cations and inorganic phosphate, citrate and chloride for the main anions. In milk, these ions are more or less associated between themselves and with proteins. Depending on the type of ion, they are diffusible (cases of sodium, potassium and chloride) or partially associated with casein molecules (cases of calcium, magnesium, phosphate and citrate), to form large colloidal particles called casein micelles. Today, our knowledge and understanding concerning this fraction is relatively complete. In this review, the different models explaining (i) the nature and distribution of these minerals (especially calcium phosphate) in both fractions of milk and (ii) their behaviour in different physico-chemical conditions, are discussed. PMID- 16045896 TI - E. coli proteolytic activity in milk and casein breakdown. AB - Previous studies have focused on both LPS and E. coli experimental mastitis and underlined the respective roles of endogenous proteolysis (including plasmin from the blood stream and other proteases from milk leukocytes), as well as the presence of E. coli in a more intricate system. The aim of this study was to assess the role of E. coli in milk proteolysis and especially that of its proteases in casein breakdown. The first part consisted in the incubation of 104 cfu.mL(-1) of the E. coli strain in raw milk at 37 degrees C for 24 h; the same milk was also incubated with 0.04% sodium azide. Several parameters were evaluated: CFU, plasmin activity, gelatinase activity and pH 4.6 insoluble peptides, including the proportion of gamma-CN. The profile of gelatinase activity was determined by zymography and identified by immunoblotting. In the second part of the study, we examined the profile of CN (alphas-, beta- and kappa CN) breakdown by E. coli lysate. The results suggest that E. coli proteases have a direct effect on CN, and the increase of gamma-CN in inoculated milk may be generated by both plasmin and the gelatinase. Moreover, the gelatinase activity in the inoculated milk was higher after 24 h of incubation. PMID- 16045897 TI - Focus on the supramolecular structure of milk fat in dairy products. AB - Bovine fat is dispersed in raw milk as natural milk fat globules, with an average diameter of 4 microm, which are enveloped in a biological membrane, the milk fat globule membrane (MFGM). However, dairy processes modify the supramolecular structure and the surface composition of milk fat. Thus, milk fat is present in many dairy products under various forms. In this study, we focused on the fact that natural milk fat globules are rarely consumed in their native state, i.e. in fresh raw milk. In most drinking milks, fat globules are homogenised in order to avoid their rising at the surface of the products. Furthermore, fat globules are heat treated to avoid the growth of micro-organisms. As a consequence of the technological process applied, the volume-weighted average diameter of fat globules in drinking milks is in the range 0.2-0.5 microm. Homogenisation of fat globules led to the partial disruption of the MFGM and to the adsorption of milk proteins. Moreover, this study showed that in cheeses, milk fat can be dispersed as (i) fat globules with the MFGM, (ii) aggregates of fat globules, (ii) homogenised fat globules, (iii) free fat and (iv) a combination of different phases and structures. The knowledge of the supramolecular structure of milk fat in dairy products is of primary importance regarding its technological, sensorial and nutritional properties. PMID- 16045898 TI - Mammary transmission of caprine arthritis encephalitis virus: a 3D model for in vitro study. AB - Transmission of Caprine Arthritis Encephalitis virus (CAEV) from the mother to offspring is principally mediated by infected cells from colostrum and milk. The infection of the dam is often sub-clinical, and results in increased cellularity of milk, sometimes exacerbated by bacterial co-infections. Although monocytes are the major viral host cells, several other cell types, including epithelial mammary cells, fibroblasts and endothelial cells show low levels of in vivo infection. In vitro, however, all phenotypes of mammary gland cells are individually highly sensitive to CAEV infection. This suggests that local mechanisms act to control viral expression. Our goal is to analyse the mechanisms regulating local virus infection, including the physiological status of the mammary gland and bacterial co-infections. In this work, we present the development of a model for the in vitro reconstitution of mammary gland tissue using 3D cultures in Matrigel. Mononuclear cells from the blood are added to the 3D cultures in vitro. In these experimental conditions, the mammary cells spontaneously organize into mammospheres. Blood leucocytes migrate into the culture gel, and localize particularly at the periphery of the mammospheres. Mammospheres were susceptible to infection in vitro by CAEV, as shown by a cytopathic effect and expression of late CAEV antigen p30. This model will allow the in vitro study of virus expression, transfer of infection to mammary gland cells and interactions between the mammary gland cells, infected monocytes and immunocompetent cells. It will allow the study of mechanisms participating in the control of passage of pathogens into milk, according to the physiological and CAEV-infection status of the animal, microenvironment and the presence of bacterial co-infections. PMID- 16045899 TI - Neurorescue effects of VEGF on a rat model of Parkinson's disease. AB - Vascular endothelial growth factor (VEGF) has been shown to display neuroprotective effects on dopaminergic (DA) neurons. Here, we investigated the neurorescue effects of VEGF on 6-hydroxydopamine (6-OHDA)-treated DA neurons in vitro and in vivo. Initially, we examined in vitro whether 1, 10, or 100 ng/ml of VEGF administration at 2 or 4 h after 6-OHDA treatment rescued DA neurons derived from E14 murine ventral mesencephalon. The earlier treatment of VEGF suppressed 6 OHDA-induced loss of DA neurons more than the delayed treatment. Next, we examined whether the continuous infusion of VEGF had neurorescue effects in a rat model of Parkinson's disease. We established a human VEGF secreting cell line (BHK-VEGF) and encapsulated the cells into hollow fibers. The encapsulated cells were unilaterally transplanted into the striatum of adult rats at 1 or 2 weeks after 6-OHDA lesions, and animals subsequently underwent behavioral and immunohistochemical evaluations. Compared to lesioned rats that received BHK Control capsules, lesioned rats transplanted with BHK-VEGF capsules showed a significant reduction in the number of amphetamine-induced rotations, a significant preservation of TH-positive neurons in the substantia nigra pars compacta, and a remarkable glial proliferation in the striatum, with the earlier transplantation exerting much more benefits than the delayed transplantation. Parallel studies revealed that the observed in vitro and in vivo neurorescue effects were likely mediated by VEGF's angiogenic and glial proliferative effects, as well as its direct effects on the neurons. Our results suggest that VEGF is a highly potent neurorescue molecule for Parkinson's disease therapy. PMID- 16045900 TI - Ketamine pre-treatment dissociates the effects of electroconvulsive stimulation on mossy fibre sprouting and cellular proliferation in the dentate gyrus. AB - Electroconvulsive stimulation (ECS), the experimental analogue of electroconvulsive therapy (ECT), has been shown to produce both functional and structural effects in the hippocampal formation in infrahuman species. These changes may relate to the antidepressant and cognitive effects of ECT observed in patients treated for severe depressive disorders. Recent studies have described both enhanced neurogenesis in the dentate gyrus of the hippocampus and sprouting of mossy fibre projections from granule cells. The behavioural significance of these effects remains uncertain. In this study, we examined whether ketamine, a clinically available non-competitive NMDA receptor channel blocker, could block both of these "trophic" effects. Rats were given a course of eight spaced ECS or sham treatments under either halothane or ketamine anaesthesia. The thymidine analogue bromodeoxyuridine was administered to assess the degree of hippocampal cell proliferation and mossy fibre sprouting was quantified using the Timm staining method. Pre-treatment with ketamine dissociated these effects such that mossy fibre sprouting was attenuated significantly, while cell proliferation was unaffected. This dissociation may prove useful in determining the behavioural significance of these hippocampal changes, if any, for either the antidepressant or cognitive consequences of ECT. PMID- 16045901 TI - Positional isomers of sulfated oligosaccharides obtained from agarans and carrageenans: preparation and capillary electrophoresis separation. AB - Partial reductive hydrolysis was used to produce oligosaccharide alditols from repetitive sulfated galactans obtained from four Rhodophyta species: kappa carrageenan (from Kappaphycus alvarezii), theta-carrageenan (Gigartina skottsbergii-alkali-treated lambda-carrageenan), agarose 6-sulfate (Gracilaria domingensis), and pyruvylated agarose 2-sulfate (Acanthophora spicifera-alkali treated pyruvylated agaran sulfate). Each hydrolyzate was submitted to anion exchange and gel-filtration chromatography, and the isolated oligosaccharide alditols were identified by 1D and 2D NMR spectroscopy and by ESI mass spectrometry. The positional isomers of the sulfated oligosaccharide alditols were then completely resolved by capillary electrophoresis in a borate buffer. Attempts to correlate the availability of the hydroxyl groups for borate complexation with the relative migration of the oligosaccharides are presented. PMID- 16045902 TI - Expression and regulation of murine macrophage angiopoietin-2. AB - Our understanding of angiogenesis has increased significantly in the past few years with the discovery of angiopoietins (Ang). Specifically, Ang2 has been associated with pathologic as well as normal vascularization. While previous studies have shown that a major source of Ang2 has been endothelial cells and tumor cells, we reasoned that macrophages would also have the ability to express angiopoietins, specifically Ang2, due to that cell's role in wound healing, tumor angiogenesis, and a number of non-oncological diseases, such as rheumatoid arthritis and psoriasis. In this study, murine macrophages constitutively expressed both transcripts and protein for Ang2 but not Ang1 or Ang3. The secretion of Ang2 was enhanced by treatment with lipopolysaccharide, interferon gamma, prostaglandin E2 and other cyclic AMP-elevating agents, as well as vascular endothelial growth factor (VEGF). Cyclic AMP-dependent protein kinase (PKA) played a major role in this enhancement since the PKA inhibitor, H89, blocked secretion of Ang2. Since stimulation of the PKA pathway can lead to macrophage production of VEGF, it is possible that enhancement of Ang2 production by macrophages may be due to autocrine responsiveness to VEGF. Adding anti-VEGF antibodies to the supernatants of stimulated macrophages blocked secretion of Ang2. This study is the first to show murine macrophage production of Ang2 and to provide evidence that it can be regulated. Understanding the regulation of macrophage Ang2 production is especially important in an effort to target the pathologic role of macrophages while preserving their role in immunity and homeostasis. PMID- 16045903 TI - Effect of inducers of DT-diaphorase on the haemolytic activity and nephrotoxicity of 2-amino-1,4-naphthoquinone in rats. AB - Reduction of naphthoquinones by DT-diaphorase is often described as a detoxification reaction. This is true for some naphthoquinone derivatives, such as alkyl and di-alkyl naphthoquinones, but the situation with other substances, such as 2-hydroxy-1,4-naphthoquinone, is more complex. In the present study, the effect of several substances that are known to increase tissue activities of DT diaphorase on the toxicity of 2-amino-1,4-naphthoquinone has been investigated. Like 2-hydroxy-1,4-naphthoquinone, the 2-amino-derivative was found to cause both haemolytic anaemia and renal tubular necrosis in rats. Again like 2-hydroxy-1,4 naphthoquinone, the severity of the haemolysis induced by the 2-amino derivative was increased in animals pre-treated with inducers of DT-diaphorase, but the degree of nephrotoxicity was decreased. With these substances, therefore, DT diaphorase both activates and detoxifies the quinone, depending on the target organ. It is not possible to generalize with regard to the effects of modulation of tissue levels of DT-diaphorase on naphthoquinone toxicity in vivo, since this may change not only the severity of the toxic effects, but also the target organ specificity. In evaluating the possible therapeutic applications of such compounds, the possibility of toxic effects upon the blood and kidney must be borne in mind. In man, renal damage by compounds such as 2-hydroxy- and 2-amino 1,4-naphthoquinone may be a particular problem, because of the low level of DT diaphorase in human liver. PMID- 16045904 TI - Angiotensinogen M235T polymorphism is associated with coronary artery disease severity. AB - INTRODUCTION: Previous reports relating coronary artery disease and functional variants of the renin-angiotensin system have been contradictory in establishing the role of these polymorphisms in coronary artery disease (CAD) development. The aim of the present study is to determine if there is an association between the M235T variant of the angiotensinogen gene and severity of coronary artery disease in patients with clinically suspected disease undergoing cineangiogram. METHODS: The angiotensinogen M235T variant was analyzed in 871 consecutive patients with clinically suspected coronary artery disease submitted to coronary angiography study. Three different angiographic scores were used to determine severity of the disease analyzing 20 coronary segments. All patients were evaluated considering classical risk factors for coronary artery disease throughout a medical oriented questionnaire, anthropometric measures, and blood glucose and lipid profile determination. RESULTS: Presence of the 235T allele was associated with higher angiographic extension scores in univariate analysis (p < 0.05). The 235T allele was also associated with an increased risk of presenting 3-vessel disease (p < 0.05). In addition, the T allele was significantly associated with higher Gensini's scores both in univariate (p = 0.05) and multivariate analyses (p < 0.05). Finally, a 1.86 fold increase in the risk of multivessel disease (95% CI: 1.174-2.947) was associated with the TT genotype independently of other cardiovascular risk factors associated with disease extension. CONCLUSION: The data hereby provide further support for the association between angiotensinogen M235T polymorphism and CAD severity independently of other cardiovascular risk factors. PMID- 16045905 TI - Characteristics and health impacts of volatile organic compounds in photocopy centers. AB - This study investigates the indoor air quality of typical photocopy centers in Taiwan to evaluate the human health risk following inhalation exposure. Both personal and area samplings were conducted at seven photocopy centers in the Tainan area from July 2002 to March 2003, which covered both summer and winter seasons in Taiwan. The benzene, toluene, ethylbenzene, xylenes, and styrene (BTEXS) measurements indicated no difference between personal and area samplings (P>0.05) and found that air conditioning improves indoor air quality. The additive factor at each photocopy center was significantly below 1.0, based on the current BTEXS permissible exposure limits in Taiwan. However, the mean benzene and styrene levels in the current study were 138 and 18 times, respectively, higher than those in another study conducted in the United States. Comparison of mass ratios of BTEXS with those of several chamber studies revealed that the photocopier is not the only volatile organic compound (VOC) source in photocopy centers. The lifetime cancer and noncancer risks for workers exposed to VOCs were also assessed. Results show that all seven centers in this study had a lifetime cancer risk exceeding 1x10(-6) (ranging from 2.5x10(-3) to 8.5x10(-5)). Regarding noncancer risk, levels of toluene, ethylbenzene, xylenes, and styrene were below the reference levels in all photocopy centers; however, the hazard indices for all still exceeded 1.0 (range 26.2-1.8) because of the high level of benzene in the photocopy centers. PMID- 16045906 TI - The role of bile duct reactive change in the pathogenesis of liver fibrosis due to hepatitis C. AB - The question addressed here is: does the bile duct reactive component of hepatitis C disease progress during the progression of the disease to cirrhosis? The question is important because if the answer to the question is yes, then an important correllated question is: does the bile duct reactive component contribute to the fibrotic change which leads to cirrhosis? The first question is addressed in the present study of a series of liver biopsies taken at the four stages of liver fibrosis in patients with hepatitis C. Sixty-four patients with hepatitis who had been biopsied for staging purposes were reviewed retrospectively. The liver biopsies were routinely stained with antibodies for liver cells, bile duct cells, activated stellate cells and cells in S phase of the cell cycle and histochemical stains for collagen and basement membrane. Selective biopsies were stained for stem cells and oval cells. There was a progressive increase in metaplastic bile ductules but the increase did not reach a significant level until stages III and IV of fibrosis. There was a positive correlation between the number of ductules formed and the stage of liver fibrosis. The incidence of proliferating metaplastic ductules was low and did not change significantly during the progression of the stage of the fibrosis. Stains for oval cells and stem cells were negative. It is concluded that the answer to the question posed is: bile ductule reaction does increase during the development of cirrhosis caused by hepatitis C but the increase is due to bile ductular metaplasia, not due to proliferation. PMID- 16045907 TI - Constitutive nitric oxide synthase activity is required to trigger ischemic tolerance in mouse retina. AB - Profound morphologic and functional protection against retinal ischemic injury can be achieved if the tissue is 'preconditioned' one day earlier with a brief, noninjurious ischemic challenge. To begin to address the mechanistic basis of this 'ischemic tolerance', we used genetic and pharmacologic approaches to test the hypothesis that nitric oxide (NO) derived from one of the three NO synthase (NOS) isoforms was responsible for triggering the adaptive response to brief preconditioning ischemia. Retinae of adult mice were preconditioned with 5-min preconditioning ischemia and subjected to 45-min injurious ischemia 24 hr later. Some animals were treated with the constitutive NOS inhibitor L-nitroarginine (5 mg/kg, i.p.) 1 hr before preconditioning. Retinal layer thicknesses and cell counts were determined one week postischemia in 5-mum thin sections, and flash electroretinograms were obtained at 1 and 7 days postischemia. We confirmed that ischemic preconditioning afforded morphologic and functional protection in the strains of wild-type mice studied. Histopathologic analyses of inducible NOS (iNOS) knockout mice revealed that ischemic preconditioning was completely effective, whereas ischemic tolerance was not achieved in the retinae of endothelial NOS (eNOS) and neuronal NOS (nNOS) knockout mice. The participation of the constitutive NOS enzymes in preconditioning-induced tolerance was confirmed by the finding that administration of the NOS inhibitor L-NA to wild type mice prior to ischemic preconditioning blocked the development of ischemic tolerance. These cross-validating genetic and pharmacologic findings indicate that NO derived from both eNOS and nNOS is a required molecular signal in the adaptive response to ischemic preconditioning in the retina. PMID- 16045908 TI - Lens epithelium supports axonal regeneration of retinal ganglion cells in a coculture model in vitro. AB - The purpose of this study was to determine whether the lens epithelium influences the survival or axonal growth of regenerating retinal ganglion cells. The optic nerves of adult albino rats were injured in order to induce axonal regeneration, and axon growth was then studied in retinal explants in the presence of cocultivated lens capsules carrying living epithelial cells. In the first series of experiments, cocultivation of retinal explants with lens epithelium in immediate proximity resulted in penetration of fibers into the lens epithelium, indicating that it supported axonal growth. In the second series of experiments, co-explants were placed 0.5-1.0mm from each other. The numbers of outgrowing retinal axons were determined both with respect to the retinal eccentricity and the topological relationship with the lenticular co-explant. The Wilcoxon matched pairs signed-rank test was used to determine if the numbers of axons differed significantly between four regions of the explants. Significantly more axons grew out from the retinal edge facing the lenticular explant than from its opposite side, indicating that the lens epithelium supports axon growth. The numbers of surviving retinal ganglion cells in culture were determined after retrograde prelabelling with a neuroanatomical tracer. The number of fluorescent ganglion cells within the retinal explants did not significantly differ between the groups (Mann-Whitney test). These findings indicate that the lens epithelium influences both the amount of axonal regeneration and the direction of growth without affecting the survival rate of retinal ganglion cells in vitro. PMID- 16045909 TI - Effects of acute delivery of endothelin-1 on retinal ganglion cell loss in the rat. AB - The vasoconstrictive peptide, Endothelin-1 (ET-1) has been found at elevated levels in glaucomatous eyes. In this study, a single 5mul intraocular injection of ET-1 was injected into the rat eye in order to characterize an in vivo retinal ganglion cell (RGC)-specific cell death model. The most effective concentration of ET-1 at inducing RGC loss at 2 weeks post-injection was determined using 5, 50 and 500mum concentrations of ET-1. The density of surviving RGCs was determined by counting Fluorogold labelled RGCs. A significant loss (25%) of RGCs was observed using only the 500mum concentration when compared to PBS-injected controls. GFAP immunohistochemistry revealed an increase in GFAP expression in Muller cell end-feet, as well as a total increase in GFAP expression (80%), following ET-1 treatment. These changes in GFAP expression are indicative of glial hyperactivity in response to stress. The specificity of ET-1 mediated cell death for RGCs was determined by measuring the changes in retinal thickness and TUNEL labeling. Retinal thickness was quantified using confocal and light microscopy. In confocal measurements, Yo Pro-1 was used to stain nuclear layers and the thickness of retinal layers determined from reconstructions. No significant loss in thickness was observed in any retinal layers. The same observations were seen in semi-thin sections when viewed by conventional transmitted light microscopy. The lack of significant thickness changes in the outer nuclear, outer plexiform or inner nuclear layer suggests that there was no significant cell loss in the retina other than in the RGC layer. Exclusive co localization of TUNEL-labelled nuclei with Fluorogold-labelled cytoplasm provided additional evidence for RGC-specific death that most likely occurs via an apoptotic mechanism. A cell death time course was performed to determine RGC loss over time. RGC losses of 25, 25, 36 and 44% were observed at 1, 2, 3 and 4 weeks post-ET-1 injection, compared to PBS-injected controls. The total number of remaining RGC axons was determined by multiplying the number of optic nerve (ON) axons per unit area, by the cross-sectional area. There was a 31% loss in total ON axons in ET-1 treated eyes at 3 weeks post injection. Functional integrity of the visual system was determined by observing changes in the pupillary light reflex. ET-1 treatment resulted in a slowing of the pupil velocity by 31% and an average increase in the duration of contraction of 1.85sec (32% increase). These experiments provide evidence that acute ET-1 injections can produce RGC-specific cell death and many cellular changes that are similar to glaucoma. This potential glaucoma model leaves the optic nerve intact and may be used in subsequent experiments, which are involved in increasing RGC survival and functional recovery. PMID- 16045910 TI - Neuronal cell loss accompanies the brain tissue response to chronically implanted silicon microelectrode arrays. AB - Implantable silicon microelectrode array technology is a useful technique for obtaining high-density, high-spatial resolution sampling of neuronal activity within the brain and holds promise for a wide range of neuroprosthetic applications. One of the limitations of the current technology is inconsistent performance in long-term applications. Although the brain tissue response is believed to be a major cause of performance degradation, the precise mechanisms that lead to failure of recordings are unknown. We observed persistent ED1 immunoreactivity around implanted silicon microelectrode arrays implanted in adult rat cortex that was accompanied by a significant reduction in nerve fiber density and nerve cell bodies in the tissue immediately surrounding the implanted silicon microelectrode arrays. Persistent ED1 up-regulation and neuronal loss was not observed in microelectrode stab controls indicating that the phenotype did not result from the initial mechanical trauma of electrode implantation, but was associated with the foreign body response. In addition, we found that explanted electrodes were covered with ED1/MAC-1 immunoreactive cells and that the cells released MCP-1 and TNF-alpha under serum-free conditions in vitro. Our findings suggest a potential new mechanism for chronic recording failure that involves neuronal cell loss, which we speculate is caused by chronic inflammation at the microelectrode brain tissue interface. PMID- 16045911 TI - Effects of lipids and aging on the neurotoxicity and neuronal loss caused by intracerebral injections of the amyloid-beta peptide in the rat. AB - The influence of diet and age on the area of lesion and on the neuronal density in the cerebral cortex was studied in rats following local injections of the amyloid-beta peptide (Abeta1-40) in PBS vehicle into the left frontal and cingulate cortices and compared with effects of injections of PBS alone into the corresponding regions of the right hemisphere The experiments were carried out in two groups of animals: one group of young adult rats and a second group of aged rats. Each group of animals, depending on the diet received, was divided into high-cholesterol, high-fat, and a control group. In order to evaluate the interaction of Abeta/PBS-cholesterol and of Abeta/PBS-fat, animals without dietary manipulation receiving Abeta and PBS injection were used as controls. The results showed that the greatest area of lesion was at Abeta injection sites in the high-cholesterol fed group of aged animals. The results also revealed a significant variance in the neuronal density by group and by injection type. Thus, high-cholesterol fed animals showed a greater reduction in neuronal density at Abeta and PBS-injected sites than that seen in the high-fat or control groups. The results also indicate that the loss of neurons at the Abeta injection site exceeds that seen in the PBS-injected area. The greatest reduction in the neuronal density was found at Abeta-injected site in the high-cholesterol fed group of aged animals. In conclusion, our findings indicate an interaction between lipids, age, and Abeta neurotoxicity, and might provide insights into the basic mechanisms involved in a short-term (acute-to-subchronic) response to Abeta peptide. PMID- 16045912 TI - Intra-sexual variation in male reproduction in teleost fish: a comparative approach. AB - The occurrence of intra-sexual variation in reproduction is a widespread phenomenon in teleosts. One such form of variation consists in the occurrence of alternative male types: males that invest resources in mate attraction and males that exploit the investment of the former males, by trying to sneak fertilizations during spawning. These alternative reproductive tactics can be classified according to their plasticity during the life span of the individuals (i.e., fixed vs. sequential vs. reversible). Furthermore, the differences between morphs within a given species may involve a set of different traits, including reproductive behavior, the differentiation of male morphological traits, and the patterns of gonad tissue allocation and the differentiation of gonadal accessory glands. In this paper, we review the available data on four species exhibiting different types of intra-sexual plasticity in reproduction that have been studied in our lab. The data on the proximate mechanisms, androgens and forebrain arginine-vasotocin (AVT), underlying these alternative tactics suggest that between-morph differences in androgen levels, especially in 11-ketotestosterone, are especially present in species where the alternative male types have evolved morphological traits that are tactic-specific (i.e., sexual polymorphisms) and that differences in AVT appear to be related to between-morph differences in the expression of courtship behavior. Therefore, this comparative approach leads us to propose that the different endocrine systems are involved in the differentiation of different sets of traits that make up alternative phenotypes, and that the differentiation of alternative tactics is not controlled by a single endocrine system (e.g., androgens). PMID- 16045913 TI - Obesity in youth is not an independent predictor of carotid IMT in adulthood. The Cardiovascular Risk in Young Finns Study. AB - Being obese in childhood may be associated with increased cardiovascular morbidity and mortality in adulthood. We examined the relationship between obesity and overweight identified in youth and carotid artery intima-media thickness assessed in adulthood. As part of the longitudinal Cardiovascular Risk in Young Finns Study, we assessed tracking of body mass index (BMI) from youth (ages 3-18 years) to young adulthood (ages 24-39 years) in a cohort of 2,260 subjects. BMI measured in youth was significantly associated with BMI measured in adulthood. The risk of being obese in adulthood (BMI > 30 kg/m(2)) was increased by three-fold in subjects who had been overweight or obese (BMI > 80th percentile) in childhood (ages 3-9 years) and by four-fold in subjects who had been overweight or obese in adolescence (ages 12-18 years). Age and sex adjusted adult IMT values were comparable in subjects who had been consistently overweight/obese in youth and adulthood and in subjects who became obese in adulthood, 0.642 mm versus 0.634 mm, respectively. IMT values were lower (overall P < 0.0001) and comparable in subjects who had remained consistently non-obese and those who had been obese in youth but had become non-obese in adulthood, 0.610 mm versus 0.627 mm, respectively. We conclude that being obese in youth is associated with increased carotid IMT in adulthood, but this relationship is explained by significant tracking of body mass from youth to adulthood. PMID- 16045914 TI - A structural basis for the aortic stress-strain relation in uniaxial tension. AB - A constitutive law that includes three analytical expressions was recently proposed to approximate the low, physiologic, and high-stress parts of the aortic stress-strain relation in uniaxial tension, consistent with the biphasic nature of the aortic wall under passive conditions. This consistency, and the fact that previous phenomenological uniaxial laws have only indirectly been related to vessel wall structure, motivates the investigation of the structural basis underlying the newly proposed three-part constitutive law. For this purpose, longitudinally oriented aortic strips were fixed in Karnovsky's solution, while subjected to various pre-selected levels of uniaxial tensile stress. Light microscopy examination disclosed that the elastic lamellae gradually unfolded at low and were almost straight at physiologic and high stresses, while collagen fibers reoriented in the longitudinal axis at low, started uncoiling at physiologic, and straightened massively at high stresses. In the circumferential sections, the elastic lamellae and the circumferentially distributed collagen bundles remained wavy at all levels of longitudinally applied stress. These microstructural changes suggest that elastin becomes load-bearing at low, and collagen at physiologic but mostly at high stresses, so that the first and third parts of the constitutive law are in turn due to the presence of elastin and collagen alone, and the second due to both elastin and collagen. The structural basis of this constitutive law allows physically significant interpretation of its parameters, offering insight into how the aortic microstructure determines the macromechanical response. PMID- 16045915 TI - Estimating propulsive forces--sink or swim? AB - The purpose of this study was to investigate the validity of hydrodynamic force estimation in swimming as calculated by the quasi-static approach. To achieve this a full-scale mechanical arm was developed, built and tested. The mechanical arm, covered with a prosthetic shell and driven at the shoulder was used to simulate a single plane underwater rotation at four elbow configurations. A computer program controlled the shoulder movement to achieve a replicable angular velocity profile for each arm movement. A strain gauge system was used to directly measure the generated arm torque. Repeated trials were conducted at fixed elbow angles of 110 degrees, 135 degrees, 160 degrees and 180 degrees. All trials were filmed using a three-dimensional underwater set-up. Each trial was digitised at 25 Hz and the hydrodynamic drag force profile of the hand calculated using the quasi-static procedure. From these data, the estimated shoulder torque was calculated and compared to the direct measurement of shoulder torque from the mechanical arm. The results showed that the arm produced a repeatable movement through the water. The shoulder torque profiles using the direct measure (the arm) and the indirect measures (quasi-static approach) differed considerably. The quasi-static approach appears not to accurately reflect the hydrodynamic force profile generated by the arm movement in swimming. Furthermore, it seems that the swimmer's hand contribution is overstated in up to date studies. It is essential that the propulsive mechanisms in swimming be further investigated if factors underpinning an optimal technique are to be established. PMID- 16045916 TI - Surface modification of sepiolite with quaternary amines. AB - This study was aimed at elucidating the mechanism of adsorption of quaternary amines, stearyldimethylbenzylammoniumchloride (SDBAC), as monomers and as micelles, distearyldimethylammoniumchloride (DDAC) and hexadecyltrimethylammoniumchloride (HTAC), on the surface of sepiolite. The adsorption capacity for these surfactants onto sepiolite, calculated by fitting the experimental data to the Langmuir-Freundlich equation, were 324% (SDBAC), 278% (DDAC), and 258% (HTAC) of the cation exchange capacity of sepiolite. The Mg(2+) ions released during the exchange process were higher than the CEC value of sepiolite because of the simultaneous dissolution of the present minerals. The water adsorption decreased with the increasing surfactant loading up to 250 mmol/kg of sepiolite, which can be ascribed to an intensification of the hydrophobic properties. With loadings above 250 mmol/kg, the water adsorption increases. Simple kinetic analysis of SDBAC adsorption was performed. The properties and the type of bonding between the surfactants and sepiolite were investigated by DT, TG, and DTG analysis. During the gradual heating in oxidizing atmosphere, the adsorbed organic material is oxidized giving rise to significant exothermic peaks. The exothermic peak temperatures in the range 200-500 degrees C depended on the surfactant loadings and provided evidence of the formation of multilayers on the sepiolite surface. PMID- 16045917 TI - Histopathological classification of lesions associated with natural paratuberculosis infection in cattle. AB - Paratuberculosis-associated lesions in 116 naturally infected adult cows, with or without clinical signs, were classified histopathologically. Tissue samples obtained focused on gut-associated lymphoid tissue. Lesions were divided into five categories. Focal lesions (n=68 cases), consisted of small granulomas in the ileal and jejunal lymph nodes or the ileocaecal lymphoid tissue. In the multifocal type (n=13 cases), small granulomas or scattered giant cells appeared in some intestinal villi, as well as in the lymph nodes. Diffuse multibacillary lesions (n=15 cases), associated with severe granulomatous enteritis affecting different intestinal locations and lymph nodes, were formed by macrophages containing large numbers of acid-fast bacilli. In diffuse lymphocytic lesions (n=3 cases), lymphocytes were the main inflammatory cells, with some macrophages or giant cells containing few if any mycobacteria. In diffuse intermediate forms (n=17 cases), the infiltrate was formed by abundant lymphocytes and macrophages, and mycobacteria were present to varying degrees related to the number of macrophages. Clinical signs and gross lesions were mainly associated with diffuse forms. Thickening of the intestinal wall, which was the most common macroscopical finding, was related to the degree of submucosal change. Mycobacterium avium subsp. paratuberculosis was identified by culture or PCR in all cows with diffuse lesions, and in 55.5 and 37% of those with multifocal or focal forms, respectively. The importance of sampling the ileal and caudal jejunal lymph nodes to find histological lesions of paratuberculosis in cattle is emphasized. PMID- 16045918 TI - Orbital meningioma with a granular cell component in a dog, with extracranial metastasis. AB - A meningioma with a significant component of granular cells is described in the left ocular orbit of a 5-year-old male Golden retriever dog that presented with exophthalmos. The neoplastic mass surrounded the optic nerve. Microscopically, the tumour was characterized by nests of large, round to polygonal cells, variable in size, with eccentric nuclei and abundant eosinophilic cytoplasm, containing PAS-positive granules. In focal areas, cells were smaller and occasionally formed whorl-like structures. Immunohistochemical analysis revealed that neoplastic cells reacted positively for vimentin and, with less intensity, for neuron specific enolase and S-100, whereas they were negative for glial fibrillary acidic protein and cytokeratins. Metastatic growths, formed by similar cells, were seen in the lung and heart. PMID- 16045919 TI - Nucleomorphometric analysis of canine cutaneous mast cell tumours. AB - Thirty-five canine cutaneous mast cell tumours (CCMCTs) were analysed by computerized nuclear morphometry. In each case, the nuclei of at least 100 neoplastic cells were measured, and the mean nuclear area (MNA), mean nuclear perimeter (MNP) and mean nuclear form factor (FF) were calculated. Significant differences in respect of MNA and MNP occurred between tumours of grades I and III and between those of grades II and III (P<0.01) but not between tumours of grades I and II (P>0.01). No significant differences in respect of FF were observed between tumours of different grades. The results obtained indicate that nuclear morphometric analysis may assist in the grading of CMCTs. PMID- 16045920 TI - Tissue distribution of phocine herpesvirus-1 (PhHV-1) in infected harbour seals (Phoca vitulina) from the central Californian coast and a comparison of diagnostic methods. AB - The polymerase chain reaction (PCR) was used to determine the tissue distribution of phocine herpesvirus-1 (PhHV-1) DNA in 20 stranded Pacific harbour seals (17 pups and three seals older than one year) that died during rehabilitation. The aim was to begin to define stages of infection and to investigate the relation between the presence of PhHV-1 in tissues, histological lesions and serology. PhHV-1 DNA was detected in a wide range of tissues from 10/17 pups and 3/3 subadults or adults. Different clinical patterns emerged from the examination of ante- and post-mortem samples. These patterns probably represented pups with active PhHV-1 infection, pups recovering from infection, and older harbour seals with chronic, reactivated infection. As PhHV-1 DNA was detected in tissues in the absence of typical histological lesions in seven seals and in the absence of PhHV 1 specific antibodies in four seals, it is clear that both histological examination and serology underestimate the presence of infection. These results showed that infection can occur in the absence of obvious disease and that seroconversion may be associated with clinical recovery. PMID- 16045921 TI - Neuroendocrine carcinoma in the nasal cavity of ten dogs. AB - Neuroendocrine (NE) carcinoma was diagnosed in 10 dogs. In six cases examined by cephalometric radiography and computerized tomography, a large mass was seen to fill the nasal cavity. Histopathologically, sheets, nests or ribbons of neoplastic cells were separated by delicate or thick fibrovascular stroma. The neoplastic cells were round, oval, or spindle-shaped; cytoplasmic granules and hyperchromatic nuclei with prominent nucleoli were present. Neoplastic cells were invariably immunohistochemically positive for cytokeratin (CK) AE1/AE3, neuron specific enolase, chromogranin A and vasoactive intestinal polypeptide. Eight dogs were positive for S100 protein, seven for synaptophysin, five for protein gene product 9.5, two for somatostatin, and one for Leu-7. Immunolabelling gave negative results for CK 8, CK 19, calcitonin, calcitonin gene-related polypeptide, neurofilaments, serotonin, gastrin and glial fibrillary acidic protein. Ultrastructurally, the neoplastic cells contained a large number of round, membrane-bounded, densely-cored granules corresponding to neurosecretory granules. These observations were consistent with the neuroendocrine nature of the carcinomas. PMID- 16045922 TI - Immuno-histochemical demonstration of the role of chlamydiaceae in renal, uterine and Salpingeal disease of the koala, and demonstration of chlamydiaceae in novel sites. AB - Numerous bacteria, including Chlamydophila pecorum and Chlamydophila pneumoniae, are known to occur in diseased sites in koalas. In the present study the significance of such organisms was investigated by demonstrating their distribution in situ, in tissues collected opportunistically from wild koalas. Chlamydiaceae were demonstrated in epithelial cells and macrophages in association with pyogranulomatous pyelonephritis (8/11 kidneys), focal interstitial nephritis (3/21), and active inflammation and fibrosis of the entire upper female reproductive tract (10/10). In one case of pyelonephritis, Gram positive cocci were also demonstrated in association with Chlamydiaceae and, in another, haematogenous filamentous bacteria appeared to be the sole aetiological agent. Three cases of chlamydial metritis were also superficially co-infected by a mixture of other bacteria. Chlamydiaceae were also demonstrated in pulmonary alveolar macrophages and epithelial cells in association with pneumonitis, and in hepatic and splenic macrophages in one koala. The study illustrated the prominent role of Chlamydiaceae in renal disease and disease of the uterus, uterine tube and ovarian bursa, with implications for pathogenesis and therapy. In addition, macrophages appeared to be a potential site of latent persistence from which systemic spread of infection might occur. PMID- 16045923 TI - Enumeration and characterization of virus-specific B cells by multicolor flow cytometry. AB - To better characterize B cell responses induced to influenza virus, we developed an assay to directly quantify and characterize virus-specific B cells. We used purified and biotinylated whole virus as well as the major influenza virus surface antigen, hemagglutinin (HA) to label virus-specific B cells induced by immunization of mice with whole influenza virus in adjuvant. Immunization with adjuvant alone caused non-specific binding of whole virus to a large number of B cells in the draining lymph nodes as assessed by flow cytometry. This precluded the use of whole virus as a specific staining reagent. In contrast, staining with bromelain-cleaved purified and biotinylated influenza virus HA identified a small population of B cells (roughly 1%) only in the draining lymph nodes of virus immunized mice. FACS-purification and subsequent ELISPOT analysis showed that HA labeled B cells contained the vast majority of virus-specific antibody-secreting cells at day 10 after immunization. Overall, virus-specific antibody-secreting cells comprised roughly 10% of the HA-labeled cells. Using HA-staining in conjunction with 8-color flow cytometry we further demonstrated that close to 90% of the HA-labeled cells were CD19+ IgD- CD23- CD24high CD38low germinal center B cells, many of which had incorporated bromodeoxyuridine, indicating recent cell division in vivo. We conclude that viral HA can be used in conjunction with cell surface and intracytoplasmic stains in multicolor flow cytometry to provide detailed phenotypic and functional information on virus HA-specific B cells. PMID- 16045924 TI - A universal strategy for stable intracellular antibodies. AB - The expression of intracellular antibodies (intrabodies) in mammalian cells has provided a powerful tool to manipulate microbial and cellular signalling pathways in a highly precise manner. However, several technical hurdles have thus far restricted their more widespread use. In particular, single-chain antibodies (scFvs) have been reported to fold poorly in the reducing environment of the cytoplasm and as such there has been a reluctance to use scFv-phage libraries as a source of intrabodies unless a preselection step was applied to identify these rare scFvs that could fold properly in the absence of disulfide bonds. Recently, we reported that scFvs can be efficiently expressed within the cytoplasm of bacteria when fused at the C-terminus of the Escherichia coli maltose-binding protein (MBP). Here, we demonstrate that such MBP-scFvs are similarly stabilized when expressed in the mammalian cell cytoplasm as well as other compartments. This was demonstrated by comparing MBP-scFv fusions to the corresponding unfused scFvs that activate a defective beta-galactosidase enzyme, others that neutralize the wild-type beta-galactosidase enzyme, and an antibody that blocks the epidermal growth factor receptor. In all cases, the MBP-scFvs significantly outperformed their unfused counterparts. Our results suggest that fusion of scFvs to MBP, and possibly to other "chaperones in the context of a fusion protein", may provide a universal approach for efficient expression of intrabodies in the mammalian cell cytoplasm. This strategy should allow investigators to bypass much of the in vitro scFv characterization that is often not predictive of in vivo intrabody function and provide a more efficient use of large native and synthetic scFv-phage libraries already in existence to identify intrabodies that will be active in vivo. PMID- 16045925 TI - Immunological methods for quantifying free and total serum IgE levels in allergy patients receiving omalizumab (Xolair) therapy. AB - Omalizumab (humanized-IgG1 anti-human IgE Fc, Xolair) complexes circulating IgE, blocking IgE binding to high affinity epsilon Fc receptors (FcepsilonR1) on mast cells and basophils. Free (non-Omalizumab bound) IgE levels in serum are a measure of effective Omalizumab dosing. The goal of this study was to quantify free (non-Omalizumab-complexed) and total serum IgE levels in asthma patients on Xolair. The concentration of (non-Omalizumab bound) free IgE in human serum was measured using a solid phase immunoenzymetric assay (IEMA) in which IgE was captured from serum with monoclonal anti-human IgE (clone HP6061) and detected with labeled-FcepsilonR1alpha. In a companion total human serum IEMA, IgE was captured from serum with the same anti-human IgE (clone HP6061) and all bound IgE was detected with labeled monoclonal anti-human IgE Fc (clone HP6029). Free and total IgE levels were quantified in pre- and 1 and 3 months post Omalizumab therapy sera from 12 allergic asthma patients. In the absence of Omalizumab, working ranges of the free and total IgE IEMAs were comparable (10-1000 kIU/l), with excellent precision, reproducibility and parallelism. Pre-Omalizumab total and free IgE levels by IEMA were highly correlated (r2=0.99, Y=0.9X+0.32, p<0.001), as were total serum IgE levels by IEMA and ImmunoCAP-250 (r2=0.98, Y=1.1X-0.05, p<0.001, n=33). In vitro reduction of free IgE (>90%) occurred at [Omalizumab:IgE] molar ratios of 2-20. Total IgE levels in 12 asthmatics increased from pre-therapy levels (52-658 kIU/l) by 1.5-5.5-fold at 1 month and 1.7-8.6 fold at 3 months of uninterrupted Omalizumab treatment. Free IgE levels fell by 49%-97% at 1 month and 45%-98% by 3 months of Omalizumab treatment. Free and total IgE levels by IEMA aid in monitoring patients receiving Omalizumab therapy. PMID- 16045926 TI - Structure of the F1-binding domain of the stator of bovine F1Fo-ATPase and how it binds an alpha-subunit. AB - The peripheral stalk of ATP synthase holds the alpha3beta3 catalytic subcomplex stationary against the torque of the rotating central stalk. In bovine mitochondria, the N-terminal domain of the oligomycin sensitivity conferral protein (OSCP-NT; residues 1-120) anchors one end of the peripheral stalk to the N-terminal tails of one or more alpha-subunits of the F1 subcomplex. Here we present the solution structure of OSCP-NT and an NMR titration study of its interaction with peptides representing N-terminal tails of F1 alpha-subunits. The structure comprises a bundle of six alpha-helices, and its interaction site contains adjoining hydrophobic surfaces of helices 1 and 5; residues in the region 1-8 of the alpha-subunit are essential for the interaction. The OSCP-NT is similar to the N-terminal domain of the delta-subunit from Escherichia coli ATP synthase (delta-NT), except that their surface charges differ (basic and acidic, respectively). As the charges of the adjacent crown regions in their alpha3beta3 complexes are similar, the OSCP-NT and delta-NT probably do not contact the crowns extensively. The N-terminal tails of alpha-subunit tails are probably alpha-helical, and so this interface, which is essential for the rotary mechanism of the enzyme, appears to consist of helix-helix interactions. PMID- 16045927 TI - Benzoquinoquinoxaline derivatives stabilize and cleave H-DNA and repress transcription downstream of a triplex-forming sequence. AB - Oligopyrimidine*oligopurine sequences with potential to form intramolecular triple helix structures (H-DNA) have been found mainly in high eukaryote genomes. However, the natural occurrence and function of H-DNA remains elusive largely because we lack appropriate reagents to demonstrate the formation of these structures in cells. We examined whether a triple-helix specific stabilizing compound, benzoquinoquinoxaline (BQQ), and its 1,10-phenanthroline derivative can be efficiently utilized to study the formation and stabilization of an intramolecular triple-helical DNA structure in growing Escherichia coli cells and in vitro. Cell uptake of BQQ was confirmed by fluorescence microscopy. A plasmid carrying an H-DNA forming sequence upstream of a reporter gene was used to assess the effects of H-DNA formation and stabilization in growing cells. The presence of the H-DNA forming sequence dramatically repressed beta-lactamase expression, and sub-growth-inhibitory doses of BQQ caused a further 40% reduction. Most importantly, repression was dependent on the triple-helix forming sequence and correlated with the addition of BQQ. As the abundance of the H-DNA forming plasmid was not affected by the addition of BQQ, the dose-dependent reduction at the protein level observed here is likely caused by repression of transcription. Finally, the triple-helix specific interaction of BQQ with the target DNA sequence was demonstrated using a triple-helix directed cleavage assay by BQQ 1,10-phenanthroline conjugate in vitro. PMID- 16045928 TI - Solution structure of human prolactin. AB - We report the solution structure of human prolactin determined by NMR spectroscopy. Our result is a significant improvement over a previous structure in terms of number and distribution of distance restraints, regularity of secondary structure, and potential energy. More significantly, the structure is sufficiently different that it leads to different conclusions regarding the mechanism of receptor activation and initiation of signal transduction. Here, we compare the structure of unbound prolactin to structures of both the homologue ovine placental lactogen and growth hormone. The structures of unbound and receptor bound prolactin/placental lactogen are similar and no noteworthy structural changes occur upon receptor binding. The observation of enhanced binding at the second receptor site when the first site is occupied has been widely interpreted to indicate conformational change induced by binding the first receptor. However, our results indicate that this enhanced binding at the second site could be due to receptor-receptor interactions or some other free energy sources rather than conformational change in the hormone. Titration of human prolactin with the extracellular domain of the human prolactin receptor was followed by NMR, gel filtration and electrophoresis. Both binary and ternary hormone-receptor complexes are clearly detectable by gel filtration and electrophoresis. The binary complex is not observable by NMR, possibly due to a dynamic equilibrium in intermediate exchange within the complex. The ternary complex of one hormone molecule bound to two receptor molecules is on the contrary readily detectable by NMR. This is in stark contrast to the widely held view that the ternary prolactin-receptor complex is only transiently formed. Thus, our results lead to improved understanding of the prolactin-prolactin receptor interaction. PMID- 16045929 TI - Nucleoporin domain topology is linked to the transport status of the nuclear pore complex. AB - Nuclear pore complexes (NPCs) facilitate macromolecular exchange between the nucleus and cytoplasm of eukaryotic cells. The vertebrate NPC is composed of approximately 30 different proteins (nucleoporins), of which around one third contain phenylalanine-glycine (FG)-repeat domains that are thought to mediate the main interaction between the NPC and soluble transport receptors. We have recently shown that the FG-repeat domain of Nup153 is flexible within the NPC, although this nucleoporin is anchored to the nuclear side of the NPC. By using domain-specific antibodies, we have now mapped the domain topology of Nup214 in Xenopus oocytes and in human somatic cells by immuno-EM. We have found that whereas Nup214 is anchored to the cytoplasmic side of the NPC via its N-terminal and central domain, its FG-repeat domain appears flexible, residing on both sides of the NPC. Moreover, the spatial distribution of the FG-repeat domains of both Nup153 and Nup214 shifts in a transport-dependent manner, suggesting that the location of FG-repeat domains within the NPC correlates with cargo/receptor interactions and that they concomitantly move with cargo through the central pore of the NPC. PMID- 16045930 TI - The periplasmic binding protein of a tripartite tricarboxylate transporter is involved in signal transduction. AB - A new type of solute importer has been identified recently in various bacterial genera and called the tripartite tricarboxylate transporter (TTT). TTTs consist of two cytoplasmic membrane proteins and a periplasmic solute-binding protein. In the whooping cough agent Bordetella pertussis, a TTT system that has been called BctCBA mediates the uptake of citrate, with BctA and BctB being the membrane components and BctC, the periplasmic protein. Here, we describe that the expression of the bctCBA operon is induced by the presence of citrate in the milieu. The signalling cascade involves both BctC and the signal transduction two component system BctDE, encoded by an operon adjacent to bctCBA. Furthermore, two hybrid analyses and affinity chromatography experiments indicated that citrate liganded BctC interacts with the periplasmic domain of the sensor protein, BctE. Thus, BctC is part of the signalling cascade leading to upregulation of the transporter operon in the presence of its solute, a new function for periplasmic binding proteins of TT transporters. PMID- 16045931 TI - Conformational differences between arrestin2 and pre-activated mutants as revealed by hydrogen exchange mass spectrometry. AB - Arrestins are regulatory proteins that bind specifically to ligand-activated phosphorylated G protein-coupled receptors to terminate G protein-mediated signaling, cause the internalization of the receptor-arrestin complex, and initiate additional intracellular signaling cascades. Multiple lines of evidence suggest that arrestin normally exists in an inactive basal state and undergoes conformational activation in the process of receptor binding. "Pre-activated" phosphorylation-independent arrestin mutants display increased binding to ligand activated but unphosphorylated receptors. The mutations are believed to expose key receptor-binding regions, allowing the mutants to mimic, to some extent, the transition of arrestin to its active state. In the present study, amide hydrogen exchange (HX) and mass spectrometry (MS) were used to examine the inactive conformation of wild-type arrestin2 and compare its solution conformation with two pre-activated mutants (R169E and 3A (I385A, V386A, F387A)). The results suggest an unexpected level of structural organization within arrestin elements containing clathrin and adaptin2-binding sites that were previously believed to be completely disordered. Increased deuterium incorporation was observed in both mutant forms compared with wild-type, indicating a change in the conformation of the mutants. Three regions demonstrated significant differences in deuterium incorporation: the first 33 residues of the N terminus and residues 243-255 (both previously implicated in receptor interaction), and residues 271-299. The results suggest that subtle differences in conformation are responsible for the significant difference in biological activity displayed by pre-activated arrestin mutants and that similar changes occur in the process of arrestin binding to the receptor. PMID- 16045932 TI - Structural dynamics of precursor and product of the RNA enzyme from the hepatitis delta virus as revealed by molecular dynamics simulations. AB - The hepatitis delta virus (HDV) ribozyme is a self-cleaving RNA enzyme involved in the replication of a human pathogen, the hepatitis delta virus. Recent crystal structures of the precursor and product of self-cleavage, together with detailed kinetic analyses, have led to hypotheses on the catalytic strategies employed by the HDV ribozyme. We report molecular dynamics (MD) simulations (approximately 120 ns total simulation time) to test the plausibility that specific conformational rearrangements are involved in catalysis. Site-specific self cleavage requires cytidine in position 75 (C75). A precursor simulation with unprotonated C75 reveals a rather weak dynamic binding of C75 in the catalytic pocket with spontaneous, transient formation of a H-bond between U-1(O2') and C75(N3). This H-bond would be required for C75 to act as the general base. Upon protonation in the precursor, C75H+ has a tendency to move towards its product location and establish a firm H-bonding network within the catalytic pocket. However, a C75H+(N3)-G1(O5') H-bond, which would be expected if C75 acted as a general acid catalyst, is not observed on the present simulation timescale. The adjacent loop L3 is relatively dynamic and may serve as a flexible structural element, possibly gated by the closing U20.G25 base-pair, to facilitate a conformational switch induced by a protonated C75H+. L3 also controls the electrostatic environment of the catalytic core, which in turn may modulate C75 base strength and metal ion binding. We find that a distant RNA tertiary interaction involving a protonated cytidine (C41) becomes unstable when left unprotonated, leading to disruptive conformational rearrangements adjacent to the catalytic core. A Na ion temporarily compensates for the loss of the protonated hydrogen bond, which is strikingly consistent with the experimentally observed synergy between low pH and high Na+ concentrations in mediating residual self cleavage of the HDV ribozyme in the absence of divalents. PMID- 16045933 TI - Road safety effects of roundabouts in Flanders. AB - INTRODUCTION: This paper analyzes the effect on road safety of 95 roundabouts that were built in Flanders between 1994 and 1999. RESULTS: The study shows that the effect on the number and severity of road accidents adjusted for the trend and regression to the mean is significant, but varies considerably in accordance with the speed limit regime on the intersection. Roundabouts are most effective on intersections of a main road with a high speed limit (90 km/h) and an adjacent road with a lower speed limit (50 or 70 km/h). The empirical analysis reveals a reduction of 34% (varying between 15% and 59%) for the total number of injury accidents, 30% (7%-45%) for light injury accidents, and 38% (27%-72%) for serious injury accidents. This study also takes a closer look at the impact of different post-implementation periods using accident data of 1-, 3-, or 6-years after the construction of a roundabout on the calculated effectiveness results and warns for a severe underestimation when a one-year period is used. IMPACT ON INDUSTRY: An effective traffic safety policy based on scientific results thus requires some patience from the policy-makers. PMID- 16045935 TI - Characterization of circulating monocytes expressing HLA-DR or CD71 and related soluble factors for 2 weeks after severe, non-thermal injury. AB - BACKGROUND: Severe injury is associated with changes in monocytes that may contribute to poor outcomes. Longitudinal characterization of monocyte response patterns after trauma may provide added insight into these immunological alterations. METHODS: Venous blood obtained seven times during post-injury days 1 through 13 from 61 patients with an injury severity score >20 was assessed by flow cytometry for monocytes (CD14+) expressing HLA-DR or CD71 (transferrin receptor) and for circulating levels of interleukin (IL) 1alpha, IL-1beta, IL-6, soluble CD14 (sCD14), tumor necrosis factor-alpha (TNF-alpha), prostaglandin E(2) (PGE(2)), thromboxane B(2) (TXB(2)), and endotoxin. Urine neopterin was measured by high-pressure liquid chromatography, expressed as a neopterin-creatinine ratio. RESULTS: Trauma patients had leucocytosis days 1 through 13, monocytosis days 5 through 13, reduced proportions of CD14+HLA-DR+ cells days 2 through 5, and elevated proportions of CD14+CD71+ cells days 1 through 13. Neopterin was elevated all days, peaking on day 10. sCD14 was elevated days 2 through 13, and there were sporadic elevations of IL-1alpha, IL-1beta, IL-6, TNF-alpha, PGE(2), TXB(2), and endotoxin. Sepsis syndrome patients (n = 6) had larger and more prolonged reductions in CD14+HLA-DR+ cells and higher neopterin values, in comparison with uneventful patient outcomes. CONCLUSIONS: Altered proportions of monocytes expressing HLA-DR and CD71 and elevated sCD14 and urine neopterin levels, for up to 2 weeks after severe injury, underscores an extended period of profound immunological effects. Additional studies to more fully assess temporal monocyte response patterns after severe injury, including activation, may be warranted. PMID- 16045936 TI - Stem cell transplantation as a therapeutic approach to organ failure. AB - BACKGROUND: Stem cell transplantation is one of the next great frontiers for surgery. Stem cells, which are undifferentiated and self-renewing, have shown the ability to differentiate into cardiomyocytes, as well as many other cell types for potential therapeutic use by surgeons. MATERIALS AND METHODS: As a result, stem cells have the potential to undo irreversible cellular damage, something traditional therapies could not cure. However, numerous issues must be resolved to permit safe and effective clinical application of stem cell therapy. These include the interpretation of cellular labeling, the origin of replicating myocytes, the homing mechanism of stem cells, and the differentiation process. RESULTS: Successful translational research will depend on precise delivery of these cells in real time to the area of interest, e.g., the spinal cord, liver, or heart. Surgeons will be better able to excise and replace/regrow, rather than excise alone. As such, a basic understanding of stem cell biology will benefit the surgeon scientist and clinical surgeon. CONCLUSIONS: The review: 1) discusses myocardial regeneration; 2) defines and categorizes stem cells; 3) presents evidence of stem cell transdifferentiation into cardiomyocytes; and, 4) delineates the therapeutic potential of stem cells in the treatment of ischemic heart disease. PMID- 16045937 TI - Possible constraints on adaptive variation in sex ratio at birth in humans and other primates. AB - There is general agreement that adaptive variation of sex ratio at birth has not been decisively demonstrated in primates (including human beings). So some workers have questioned whether it actually exists. Others have conjectured that it exists but is subject to as yet unidentified 'constraints' (factors opposing the modifying influences of selection in the phenotype). Meanwhile though most workers have called for research to reveal the proximate causes of sex ratio variation, few (if any) have directed studies toward that end. Here it is argued that hormonal action is responsible both for much adaptive and non-adaptive sex ratio variation, and for constraints on the adaptive variation. My hypothesis proposes that levels of steroid hormones (testosterone and oestrogen) of both parents around the time of conception are positively associated with offspring sex ratio (proportion male at birth) of mammals including man. Testosterone in men and oestrogen in women are also known to be positively associated with the health, attractiveness and fertility of individual human beings. However, high levels of testosterone in women are frequently associated with adverse medical conditions. It is suggested that for these reasons (and contrary to some adaptive theory) some classes of people (particularly women) in suboptimal health ("condition") produce excesses of sons. It seems that gonadal hormones are responsible for adaptive variation; and that maternal adrenal hormones are responsible for maladaptive variation. In evolutionary terms, gonadal hormones precede adrenal hormones. PMID- 16045938 TI - Influence of caste polyethism on longevity of workers in social insect colonies. AB - Different patterns of division of labor can affect the expected longevity of social insects workers. It has been earlier suggested that when tasks performed inside and outside colony are equally risky then the expected longevity of workers in colonies with caste polyethism is greater than that in colonies without polyethism. To verify these predictions I used a model assuming two sets of tasks, associated with different mortality rates. In the colony without polyethism the workers preformed safe and risky tasks in turn, while in the colony with caste polyethism the workers specialized in only one set of tasks. The outcomes suggest that the expected longevity of workers in colonies with caste polyethism cannot be greater than that in colonies without polyethism. Only if there is no aging and under some special and rare conditions are there no differences in expected longevity between colonies with and without caste polyethism. If aging is independent of activity, caste polyethism does not shorten longevity when all tasks in the colony are equally risky. The results can explain why caste polyethism is not as widespread among social insects as age polyethism. PMID- 16045939 TI - Signal transduction networks: topology, response and biochemical processes. AB - Conventionally, biological signal transduction networks are analysed using experimental and theoretical methods to describe specific protein components, interactions, and biochemical processes and to model network behavior under various conditions. While these studies provide crucial information on specific networks, this information is not easily converted to a broader understanding of signal transduction systems. Here, using a specific model of protein interaction we analyse small network topologies to understand their response and general properties. In particular, we catalogue the response for all possible topologies of a given network size to generate a response distribution, analyse the effects of specific biochemical processes on this distribution, and analyse the robustness and diversity of responses with respect to internal fluctuations or mutations in the network. The results show that even three- and four-protein networks are capable of creating diverse and biologically relevant responses, that the distribution of response types changes drastically as a function of biochemical processes at protein level, and that certain topologies strongly pre dispose a specific response type while others allow for diverse types of responses. This study sheds light on the response types and properties that could be expected from signal transduction networks, provides possible explanations for the role of certain biochemical processes in signal transduction and suggests novel approaches to interfere with signaling pathways at the molecular level. Furthermore it shows that network topology plays a key role on determining response type and properties and that proper representation of network topology is crucial to discover and understand so-called building blocks of large networks. PMID- 16045940 TI - Some properties of the speciation model for food-web structure-mechanisms for degree distributions and intervality. AB - We present a mathematical analysis of the speciation model for food-web structure, which had in previous work been shown to yield a good description of empirical data of food-web topology. The degree distributions of the network are derived. Properties of the speciation model are compared to those of other models that successfully describe empirical data. It is argued that the speciation model unifies the underlying ideas of previous theories. In particular, it offers a mechanistic explanation for the success of the niche model of Williams and Martinez and the frequent observation of intervality in empirical food webs. PMID- 16045941 TI - The evolution of n-player cooperation-threshold games and ESS bifurcations. AB - An evolutionary game of individuals cooperating to obtain a collective benefit is here modelled as an n-player Prisoner's Dilemma game. With reference to biological situations, such as group foraging, we introduce a threshold condition in the number of cooperators required to obtain the collective benefit. In the simplest version, a three-player game, complex behaviour appears as the replicator dynamics exhibits a catastrophic event separating a parameter region allowing for coexistence of cooperators and defectors and a region of pure defection. Cooperation emerges through an ESS bifurcation, and cooperators only thrive beyond a critical point in cost-benefit space. Moreover, a repelling fixed point of the dynamics acts as a barrier to the introduction of cooperation in defecting populations. The results illustrate the qualitative difference between two-player games and multiple player games and thus the limitations to the generality of conclusions from two-player games. We present a procedure to find the evolutionarily stable strategies in any n-player game with cost and benefit depending on the number of cooperators. This was previously done by Motro [1991. Co-operation and defection: playing the field and the ESS. J. Theor. Biol. 151, 145-154] in the special cases of convex and concave benefit functions and constant cost. PMID- 16045942 TI - Inorganic nitrogen control in wastewater treatment ponds from a fish farm (Orbetello, Italy): denitrification versus Ulva uptake. AB - The aim of this study was to quantify the N removal efficiency of an Ulva-based phytotreatment system receiving wastewaters from a land-based fish farm (Orbetello, Italy), to identify the main biogeochemical pathways involved and to provide basic guidelines for treatment implementation and management. Fluxes of O2 and nutrients in bare and in Ulva colonised sediments were assessed by light/dark core incubations; denitrification by the isotope pairing technique and Ulva growth by in situ incubation of macroalgal disks in cages. O2 and nutrient budgets were estimated as sum of individual processes and further verified by 24 h investigations of overall inlet and outlet loads. Ulva uptake (up to 7.8 mmol Nm(-2) h(-1)) represented a net sink for water column and regenerated NH4+ whilst N removal via denitrification (10-170 micromol Nm(-2) h(-1)) accounted for a small percentage of inorganic nitrogen load (<5%). Laboratory experiments demonstrated a high potential for denitrification (over 800 microM Nm(-2) h(-1)) indicating that N loss could be enhanced. The control of Ulva standing stocks by optimised harvesting of surplus biomass may represent an effective strategy to maximise DIN removal and could result in the assimilation of approximately 50% of produced inorganic nitrogen. PMID- 16045943 TI - Pesticide contamination of the coastline of Martinique. AB - In January and February 2002, the presence of certain agricultural pesticides throughout the coastline of the Caribbean island of Martinique was investigated. The tropical climate of the French West Indies is suitable for banana production, which requires intensive use of pesticides. An inventory of all pesticides used on the island (compounds and tonnage) was compiled. Surveys and analyses revealed the presence of pesticides in the plumes of seven rivers. The organochlorine chlordecone and metabolites of aldicarb were detected at nearly all of the monitored sites, even though the use of chlordecone has been prohibited since 1993. Two triazines (ametryn and simazine) were also identified. The concentrations of carbamates and triazines detected in the water and sediment samples from Martinique are comparable to those reported for mainland France. Chlordecone concentrations in the sediment and particulate matter samples were, however, particularly high in the samples from Martinique. Toxicological implications are discussed. Of particular concern are the high levels of chlordecone (which is bioaccumulating and carcinogenic) and further monitoring of this compound is recommended, especially in fish and other sea-food products. PMID- 16045944 TI - Attentional orienting triggered by gaze in schizophrenia. AB - The ability to detect the direction of another person's gaze and to shift our own attention reflexively in the same direction facilitates the sharing of attention with other people. Such sharing of attention would seem critical for the maintenance of normal social cognition. Social cognition is severely impaired in people with schizophrenia. So, we used spatial cuing paradigms to investigate reflexive (Experiment 1) and controlled (Experiment 2) attentional orienting triggered by gaze in schizophrenia. In Experiment 1, 30 patients and 24 controls detected targets appearing right or left of a central image of a head turned right, left, or straight-ahead. These gaze-cues were non-predictive. Patients, but not controls, showed a significant congruency advantage at 100 ms SOA. The congruency advantage was similar in patients and controls at 300-800 ms SOA. In Experiment 2, 20 patients and 24 controls detected targets 300-800 ms after a central gaze-cue that pointed away from the target on 80% of trials. Controls, but not patients, were able to reverse the reflexive congruency advantage at 800 ms SOA. This study provides the first evidence that people with schizophrenia show abnormally sensitive gaze-triggered reflexive orienting. Findings are discussed in light of recent neuroimaging work investigating the neural basis of social orienting and social cognition. PMID- 16045945 TI - Dietary restriction alters fine motor function in rats. AB - A number of standard behavioral tasks in animal research utilize food rewards for positive reinforcement. In order to enhance the motivation to participate in these tasks, animals are usually placed on a restricted diet. While dietary restriction (DR) has been shown to have beneficial effects on recovery after brain injury, life span and aging processes, it might also represent a stressor. Since stress can influence a broad range of behaviors, the purpose of this study was to assess whether DR may have similar effects on skilled movement. Adult male Long-Evans rats were trained and tested in a skilled reaching task both prior to and during a mild food restriction regimen that maintained their body weights at 90-95% of baseline weight for eight days. The observations revealed that DR decreased reaching success and increased the number of attempts to grasp a single food pellet. The animals appeared to be more frantic when attempting to reach for food pellets, and the time taken to reach for 20 pellets decreased following the onset of DR. A second experiment investigating behaviors that do not require food rewards, including a ladder rung walking task and an open field test, confirmed that rats on DR display deficits in skilled movements and are hyperactive. These findings suggest that results obtained in motor tasks using food rewards need to be interpreted with caution. The findings are discussed with respect to stress associated with DR. PMID- 16045946 TI - Acylated peonidin glycosides from duskish mutant flowers of Ipomoea nil. AB - Five acylated peonidin glycosides were isolated from the pale gray-purple flowers of a duskish mutant in the Japanese morning glory (Ipomoea nil or Pharbitis nil) as major pigments, along with a known anthocyanin, Heavenly Blue Anthocyanin (HBA). Three of these were based on peonidin 3-sophoroside and two on peonidin 3 sophoroside-5-glucoside as their deacylanthocyanins; both deacylanthocyanins were acylated with caffeic acid and/or glucosylcaffeic acids. By spectroscopic and chemical methods, the structures of the former three pigments were determined to be 3-O-[2-O-(6-O-(trans-caffeoyl)-beta-D-glucopyranosyl)-beta-D-glucopyranoside], 3-O-[2-O-(6-O-(3-O-(beta-D-glucopyranosyl)-trans-caffeoyl)-beta-D-glucopyranosyl) 6-O-(4-O-(6-O-(3-O-(beta-D-glucopyranosyl)-trans-caffeoyl)-beta-D-glucopyranosyl) trans-caffeoyl)-beta-glucopyranoside], and 3-O-[2-O-(6-O-(trans-caffeoyl)-beta-D glucopyranosyl)-6-O-(4-O-(6-O-(3-O-(beta-D-glucopyranosyl)-trans-caffeoyl)-beta-D glucopyranosyl)-trans-caffeoyl)-beta-D-glucopyranoside] of peonidin. The structures of the latter two pigments were also confirmed as 3-O-[2-O-(6-O-(trans caffeoyl)-beta-D-glucopyranosyl)-beta-D-glucopyranoside]-5-O-beta-D glucopyranoside, and 3-O-[2-O-(6-O-(trans-caffeoyl)-beta-D-glucopyranosyl)-6-O-(4 O-(6-O-(3-O-(beta-D-glucopyranosyl)-trans-caffeoyl)-beta-D-glucopyranosyl)-trans caffeoyl)-beta-D-glucopyranoside]-5-O-beta-D-glucopyranoside of peonidin. The mutation affecting glycosylation and acylation in anthocyanin biosynthesis of Japanese morning glory was discussed. PMID- 16045947 TI - Composition of fatty acids triacylglycerols and unsaponifiable matter in Calophyllum calaba L. oil from Guadeloupe. AB - The composition of the kernel oils of two Calophyllum species (Calophyllum calaba L. and Calophyllum inophyllum L.) was investigated. The physico-chemical properties and fatty acid composition of the kernel oils were examined. In two species, oleic acid C18:1 (39.1-50%) is the dominating fatty acid followed by linoleic acid C18:2 (21.7-31.1%) as the second major fatty acid. Stearic C18:0 (13.4-14.3%) and palmitic C16:0 (11-13.7%) acids are the major saturates. The oils contains an appreciable amount of unsaturated fatty acids (70.8-73.10%). Most of the fatty acids are present as triacylglycerol (76.7-84%), twenty one triacylglycerols are detected with predominantly unsaturated triacylglycerols. The total unsaponifiable content, its general composition and the identity of the components of the sterol and tocopherol fractions are presented. In both species, analysis of the unsaponifiable fractions revealed the preponderance of phytosterols, mainly stigmasterol (35.8-45.1%) and beta-sitosterol (41.1-43.1%). Among the eight tocopherols and tocotrienols present in two species, variations exist; alpha-tocopherol (183 mg/kg) is the main tocopherol in Calophyllum calaba L. and Delta-tocotrienol (236 mg/kg) is the dominant tocotrienol in Calophyllum inophyllum L. PMID- 16045948 TI - The Shetland Islands scrapie monitoring and control programme: analysis of the clinical data collected from 772 scrapie suspects 1985-1997. AB - There were 574 scrapie positive suspects (histopathological scrapie lesions present) and 198 scrapie negative suspects (histopathological scrapie lesions absent). The greatest number of scrapie cases were recorded in sheep of 2, 3 and 4 years of age which represented 17%, 36% and 23% of the scrapie positive suspects, respectively. The sign sensitivities and specificities for the ten recorded signs were, respectively: pruritus (62%, 42%), ataxia (23%, 74%), hyperaesthesia (32%, 74%), wool loss (25%, 73%), fleece discolouration (29%, 85%), bruxism (23%, 69%), nibbling reflex (17%, 58%), head rubbing (47%, 78%), poll rubbing (25%, 83%). These single signs had poor discriminatory values with likelihood ratios close to one (range 0.89-1.21); combinations of the four signs, pruritus, wool loss, ataxia, hyperaesthesia and emaciation were more discriminatory (range 0.30-4.3). This study covered a time period when bovine spongiform encephalopathy (BSE) might have been introduced into the sheep population on the Shetland Islands via contaminated feed. No temporal changes could be detected in the age structure of the affected animals. PMID- 16045949 TI - An epidemiological evaluation of Mycobacterium bovis infections in wild game animals of the Spanish Mediterranean ecosystem. AB - Recreational hunting of indigenous wild artiodactyls has been one of the most lucrative and rapidly growing industries in Western Spain over the last five years. In the absence of careful ecological management, one consequence of the commercial exploitation of this natural resource has been the appearance of outbreaks of infectious disease; most notably bovine tuberculosis. From the outset of the study in 1997, we have observed a steady increase in prevalence of Mycobacterium bovis (M. bovis) in both species reaching 1.74 (+/-0.17) in deer in 2002 and 2.32 (+/-0.24) in wild boar. The latter species seems to be most severely affected with pulmonary lesions appearing more chronic than those observed in deer. In this study, we describe the epidemiology of M. bovis in European wild boar (Sus scrofa) and Iberian red deer (Cervus elaphus hispanicus) in Extremadura (W. Spain); a region where there are large areas of natural habitat for these species. PMID- 16045950 TI - PCR detection of colostrum-associated Maedi-Visna virus (MVV) infection and relationship with ELISA-antibody status in lambs. AB - A recent large-scale experimental study showed that bottle-feeding ovine colostrum from seropositive ewes results in high MVV-seroconversion in lambs. In contrast, relatively few lambs that naturally suckled colostrum from seropositive dams seroconverted as a result of it. Furthermore, lambs fed uninfected bovine colostrum readily seroconverted when mixed with ovine-colostrum lambs indicating that horizontal MVV transmission between lambs was efficient. MVV-infection was further investigated in the same samples using two PCR tests targeting sequences in the long-terminal repeats (LTR) and POL MVV genes. PCR-tests confirmed previous serological findings. However, the LTR-PCR was more sensitive and allowed detecting infection earlier than the other tests, including 5-8% of new born lambs from seropositive dams, providing more evidence that prenatal MVV infection may be more important than considered. The degree of agreement between PCR and antibody tests in individual samples was low up to 6 months of age and moderate at 10 months-old. Nine percent of lambs were always PCR-negative but seroconverted and 19% of lambs were PCR-positive at least once and did not seroconvert. However, seroconversion was associated with increasing number of times lambs were PCR-positive and ovine colostrum-fed lambs were more frequently PCR-positive than other lambs. The significance of these findings in terms of MVV infection, epidemiology and control is discussed. PMID- 16045951 TI - Benzothieno and benzofurano annelated estranes. AB - The preparation of estrone derived benzothieno- and benzofurano fused steroids is described. Keystep is an intramolecular thienyl(/furyl)-ene-yne cyclization of 16 ethynyl-17-heterarylestra-1,3,5(10),16-tetraenes. The cyclization was carried out under Pt as well as under Ru catalysis. PMID- 16045952 TI - Pharmacokinetics of prednisone and prednisolone in a case of hypothyroidism: effect of replacement therapy. AB - We found this particular case during the course of a clinical trial designed to assess the pharmacokinetics of oral prednisone in normal and diseased children. The plasma concentrations of prednisone, its main metabolite prednisolone, and endogenous cortisol were measured by HPLC at selected times during 8-h periods starting at 7:30 a.m. One 9.9-year-old administered prednisone 0.5mg/kg p.o. was found to be hypothyroid (TSH: 351microIU/mL; fT4: <2pg/mL; fT3: <1pg/mL); four age-matched normal boys (aged 6.6+/-4.9 years) served as a control group. In comparison with the controls, the hypothyroid boy showed a marked increase in the total AUC of prednisone (3360microg h/L versus 215+/-83microg h/L) and prednisolone (4040microg h/L versus 724+/-77microg h/L), and an altered pattern of endogenous cortisol, which is known to be impaired in hypothyroid subjects. After 6 months of thyroxine replacement therapy (75microg/day), the AUCs of prednisone and prednisolone returned to normal values (prednisone: 248microg h/L; prednisolone: 528microg h/L), as did the pattern of circadian cortisol secretion. In conclusion, our data indicate that the pharmacokinetics of prednisone and prednisolone can be profoundly altered by hypothyroidism, and subsequently restored by thyroxine replacement therapy. PMID- 16045953 TI - Modification of N-Methyl-N-Nitrosourea initiated bladder carcinogenesis in Wistar rats by terephthalic acid. AB - The effect of terephthalic acid (TPA) on urinary bladder carcinogenesis was examined. Male Wistar rats were initiated by injection of N-Methyl-N-Nitrosourea (MNU) (20 mg/kg b.w. ip) twice a week for 4 weeks, then given basal diet containing 5% TPA, 5% TPA plus 4% Sodium bicarbonate (NaHCO3) or 1% TPA for the next 22 weeks, and then euthanized. 5% TPA treatment induced a high incidence of urinary bladder calculi and a large amount of precipitate. Though 5% TPA plus 4% Sodium bicarbonate (NaHCO3) and 1% TPA treatment did not induce urinary bladder calculi formation, they resulted in a moderate increase in urinary precipitate. Histological examination of urinary bladder revealed that MNU-5% TPA treatment resulted in a higher incidence of simple hyperplasia, papillary or nodular hyperplasia (PN hyperplasia), papilloma and cancer than MNU control. MNU-5% TPA plus 4% Sodium bicarbonate (NaHCO3) and 1% TPA treatment increased slightly the incidence of simple hyperplasia and PN hyperplasia (not statistically significant). The major elements of the precipitate are phosphorus, potassium, sulfur, chloride, calcium and TPA. The present study indicated that the calculi induced by TPA had a strong promoting activity on urinary bladder carcinogenesis and the precipitate containing calcium terephthalate (CaTPA) may also have weak promoting activity on urinary bladder carcinogenesis. PMID- 16045954 TI - In silico prediction of harmful effects triggered by drugs and chemicals. AB - While the computer-assisted discovery and optimization of drug candidates based on the known three-dimensional structure of the macromolecular target (structure based design) or a binding-site surrogate (receptor modeling) is doubtless one of the more potent approaches in rational drug design, the simulation and quantification of side effects triggered by drugs and chemicals are still in their infancy. Major obstacles include the often not available 3D structure of the molecular target, the low specificity of the involved bioregulators and the identification of the controlling metabolic pathways. In the recent past, our laboratory has explored concepts allowing to simulate receptor-mediated toxic phenomena by developing algorithms, allowing to construct realistic 3D binding site surrogates of receptors known or assumed triggering adverse effects and validating them against large batches of molecular data. The underlying technology (software Quasar and Raptor, respectively) specifically allows for induced fit, solvation phenomena and entropic effects. It has been applied to various systems both of pharmacological and toxicological interest including the neurokinin-1, chemokine-3, bradykinin B(2), steroid, 5 HT(2A), aryl hydrocarbon, estrogen and androgen receptor, respectively. In this account, we describe the design of a virtual laboratory allowing for a reliable estimation of harmful effects triggered by drugs, chemicals and their metabolites in silico. In the recent past, the Biographics Laboratory 3R has compiled a 3D database including the surrogates of three major receptor systems known to mediate adverse effects (the aryl hydrocarbon, the estrogen and the androgen receptor, respectively) and validated them against a total of 345 compounds (drugs, chemicals, toxins) using multidimensional QSAR technologies. Within this pilot project, we could demonstrate that our virtual laboratory is able to both recognize toxic compounds substantially different from those used in the training set as well as to classify harmless compounds as being nontoxic. This suggests that our approach may be used for the prediction of adverse effects of drug molecules and chemicals. It is the aim to provide cost-covering access to this technology- particularly to universities, hospitals and regulatory bodies--as it bears a significant potential to recognize hazardous compounds early in the development process and hence improve resource and waste management as well as reduce animal testing. The Biographics Laboratory 3R is a non-profit-oriented organization aimed at reducing animal experimentation in the biomedical sciences by computational approaches (cf. http://www.biograf.ch). PMID- 16045955 TI - Electrostatic interactions govern both nucleation and elongation during phage P22 procapsid assembly. AB - Icosahedral capsid assembly is an example of a reaction controlled solely by the interactions of the proteins involved. Bacteriophage P22 procapsids can be assembled in vitro by mixing coat and scaffolding proteins in a nucleation limited reaction, where scaffolding protein directs the proper assembly of coat protein. Here, we investigated the effect of the buffer composition on the interactions necessary for capsid assembly. Different concentrations of various salts, chosen to follow the electroselectivity series for anions, were added to the assembly reaction. The concentration and type of salt was found to be crucial for proper nucleation of procapsids. Nucleation in low salt concentrations readily occurred but led to bowl-like partial procapsids, as visualized by negative stain electron microscopy. The edge of the partial capsids remained assembly-competent since coat protein addition triggered procapsid completion. The addition of salt to the partial capsids also caused procapsid completion. In addition, each salt affected both assembly rates and the extent of procapsid formation. We hypothesize that low salt conditions increase the coat protein:scaffolding protein affinity, causing excessive nuclei to form, which decreases coat protein levels leading to incomplete assembly. PMID- 16045956 TI - Competition in bistable vision is attribute-specific. AB - We employ ambiguous figures and rivalrous stimuli that have multiple ambiguous properties to show that the different attributes of an ambiguous stimulus can undergo independent switching dynamics. This suggests that competition is distributed and attribute-specific, consistent with the known functional segregation of visual processing. Conflicting evidence that binocular rivalry is an early or late visual process may be better understood as evidence for attribute-specific competition occurring at multiple stages of visual processing. Specifically, we show that whether perceptual selection during binocular rivalry is early and eye-based or late and percept-based depends on the particular ambiguous attributes of the rivalrous stimulus. PMID- 16045957 TI - Irrepressible saccades from a tectal lesion in a Rhesus monkey. AB - We present a case of spontaneously occurring irrepressible saccades in an experimental Rhesus monkey. Though eye jerks are sometimes associated with cerebellar disease, central demyelination or brainstem lesions, there is little consensus on their neurological mechanisms. From neurological and anatomical investigation we report that these irrepressible saccades were caused by a discrete cerebrovascular accident that involved the rostral superior colliculus along with its commissure, and with minor invasion of periaqueductal gray and adjacent mesencephalic reticular formation. Other suspected structures, like the raphe interpositus, substantia nigra and the cerebellum, were unaffected. PMID- 16045958 TI - Bistable Glass-pattern motion reveals two different processes. AB - Examining the two motion processes is an elusive task due to the difficulty of finding a proper stimulus paradigm. A rotational Glass pattern created with a random-dot array by superimposing its rotated version on the top of it can provide such a paradigm. If we displace only its rotated part in the vertical or horizontal direction, a bistable motion occurs; local dot motion in the same direction and Glass-pattern motion in the orthogonal direction. From two experiments, we found local dot motion is predominant in short spatiotemporal range and global pattern motion in long spatiotemporal range. Since the stimulus allows us to maintain all of its properties identical except for the changes in spatiotemporal parameters, this result shows more robustly that the energy-based first-order motion favors short spatiotemporal ranges while the pattern-based second-order motion favors long spatiotemporal ranges. PMID- 16045959 TI - Effect of myopia on visual acuity measured with laser interference fringes. AB - The aim was to determine how visual acuity is affected by myopia when optical factors of the eye are controlled. Grating acuity was measured with interference fringes to avoid the effects of aberrations, and ocular biometry was used to compensate for differences in retinal image size among subjects. Distance spectacle refractions ranged from +2.25 to -14.75 D. The retinal magnification factor (RMF) in mm/deg was computed for each eye from the distance refraction, central corneal power and ultrasound biometry. A forced-choice orientation discrimination method was used to measure acuity for high-contrast 543 nm laser interference fringes in three retinal locations: the fovea, and at 4 deg and 10 deg eccentricity in the temporal retina. Acuity, expressed in c/deg and adjusted for spectacle magnification, was not significantly correlated with refraction at any of the three retinal locations. When acuity was converted to retinal spatial frequency units (c/mm) via the RMF, acuity decreased with increasing myopia at all three retinal locations (significantly at the fovea and at 10 deg eccentricity). Retinal acuity values in highly myopic subjects (>6 D) are consistent with retinal sampling distances that are larger than published values of human cone or ganglion cell spacing. The results imply that a highly myopic eye has retinal neurons that are more widely spaced than normal, but the increased axial length enlarges the retinal image enough to compensate for the retinal stretching. The data are consistent with a retinal stretching model that primarily affects the posterior pole. PMID- 16045960 TI - Dynamic control of ocular disaccommodation: first and second-order dynamics. AB - Velocity and acceleration characteristics provide valuable information about dynamic control of accommodation. We investigated velocity and acceleration of disaccommodation (near-far focusing) from three starting positions. Peak velocity and peak acceleration of disaccommodation increased with the proximity of starting position however for a given starting position they were invariant of response magnitude. These results suggest that all disaccommodation responses are initiated towards a constant primary destination and are switched mid-flight to attain the desired final position. Large discrepancies between the primary destination and desired final position appear to produce overshoots and oscillations of small responses from proximal starting positions. PMID- 16045961 TI - Effects of global shape on angle discrimination. AB - Previous studies have been inconclusive as to whether angle discrimination performance can be predicted by the sensitivity of orientation discrimination mechanisms or by that of mechanisms specialised for angle coding. However, these studies have assumed that angle discrimination is independent of the shape of the object of which the angle is a part. This assumption was tested by measuring angle discrimination using angles that were parts of different triangular shapes. Angle discrimination thresholds were lowest when angles were presented in isosceles triangles (sides forming the angle were of identical length). Performance was significantly poorer when angles were presented in scalene triangles (sides of different lengths) and as much as three times worse when the sides forming the angle varied randomly in length between presentations. Comparing orientation discrimination for single lines with angle discrimination for different stimulus conditions (isosceles, scalene and random triangles) leads to conflicting conclusions as to the mechanisms underlying angle perception: line orientation sensitivity correctly predicts angle discrimination for random triangles, but underestimates angle acuity for isosceles triangles. The fact that performance in angle discrimination tasks is strongly dependant on the overall stimulus geometry implies that geometric angles are computed by mechanisms that are sensitive to global aspects of the stimulus. PMID- 16045962 TI - Estimating the dual-enzyme kinetic parameters for Cr (VI) reduction by Shewanella oneidensis MR-1 from soil column experiments. AB - Hexavalent chromium (Cr (VI)) contamination of soil and groundwater is considered a major environmental concern. Bioreduction of Cr (VI) to trivalent chromium (Cr (III)) can be considered an effective technology in remediating Cr (VI) contaminated sites. Among the Cr (VI) reducing bacteria, Shewanella oneidensis MR 1 (MR-1) is relatively effective. Reduction of Cr (VI) by MR-1 is defined by the dual-enzyme kinetic model. The feasibility of bioreduction of Cr (VI) is usually evaluated by performing batch experiments which may not accurately represent the subsurface environment. The objective of this paper is to present a rational approach to determine the dual-enzyme reaction kinetics of Cr (VI) reduction by MR-1 from continuous flow soil column experiments which more accurately simulate the subsurface environment. Kinetic parameters obtained from soil column data were found to be much smaller than that obtained from batch data. The stable enzyme induced reaction rate was found to be 127th and the average deactivating enzyme induced reaction rate was 117th of the corresponding values obtained from batch reactor data as reported in literature. PMID- 16045963 TI - Modeling monochloramine loss in the presence of natural organic matter. AB - A comprehensive model describing monochloramine loss in the presence of natural organic matter (NOM) is presented. The model incorporates simultaneous monochloramine autodecomposition and reaction pathways resulting in NOM oxidation. These competing pathways were resolved numerically using an iterative process evaluating hypothesized reactions describing NOM oxidation by monochloramine under various experimental conditions. The reaction of monochloramine with NOM was described as biphasic using four NOM specific reaction parameters. NOM pathway 1 involves a direct reaction of monochloramine with NOM (k(doc1) = 1.05 x 10(4)-3.45 x 10(4) M(-1) h(-1)). NOM pathway 2 is slower in terms of monochloramine loss and attributable to free chlorine (HOCl) derived from monochloramine hydrolysis (k(doc2) = 5.72 x 10(5)-6.98 x 10(5) M(-1) h(-1)), which accounted for the majority of monochloramine loss. Also, the free chlorine reactive site fraction in the NOM structure was found to correlate to specific ultraviolet absorbance at 280 nm (SUVA280). Modeling monochloramine loss allowed for insight into disinfectant reaction pathways involving NOM oxidation. This knowledge is of value in assessing monochloramine stability in distribution systems and reaction pathways leading to disinfection by-product (DBP) formation. PMID- 16045964 TI - Effect of EDTA and Fe-EDTA complex concentration on TCF Kraft mill effluent degradability. Batch and continuous treatments. AB - The effect of ethylenediaminetetracetic acid (EDTA) and Fe-EDTA complex on synthetic totally chlorine-free (TCF) effluent degradability in batch and continuously operating reactors was evaluated. Under batch treatment, the addition of EDTA and Fe-EDTA complex was studied in the range of 80 to 320 mg l( 1). Under continuously operated reactors, the Fe-EDTA complex concentration varied from 20 to 80 mg l(-1), and the hydraulic retention time (HRT) varied from 48 to 24 h. Sludge oxygen uptake rate (OUR) and chemical oxygen demand (COD) removal decreased when EDTA concentration increased in the influent under batch treatment; however, this inhibitory effect was reduced by the addition of Fe-EDTA complex. Without the addition of EDTA, COD removal decreased from 71% to 8%. The most efficient EDTA removal treatment (almost 10%) was the treatment of 80 mg l( 1) Fe-EDTA. Under continuously operated reactors, COD removal was greater than 57% in the synthetic TCF effluent with a Fe-EDTA concentration that varied from 20 to 80 mg l(-1); however, EDTA removal was lower than 25% in all cases. Synthetic TCF effluent with a Fe -EDTA concentration higher than 80 mg l(-1) could not be treated by the activated sludge treatment due to EDTA's inhibitory effect on the sludge. PMID- 16045965 TI - Enhanced post-denitrification without addition of an external carbon source in membrane bioreactors. AB - This study investigates a post-denitrification process without the addition of an external carbon source combined with an enhanced biological phosphorus removal (EBPR) in a membrane bioreactor (MBR). Three trial plants, with two different process configurations, were operated on two different sites, and a variety of accompanying batch tests were conducted. It was shown that even without dosing of an external carbon source, denitrification rates (DNR) much above endogenous rates could be obtained in post-denitrification systems. Furthermore, the anaerobic reactor located ahead of the process had a positive impact on the DNR. Given these surprising results, the project team decided to identify the carbon source used by the microorganisms in the post-denitrification process. Batch tests could demonstrate that lysis products do not play a major role as a C source for post-denitrification. The following hypothesis was proposed to explain the observations: the glycogen, internally stored by the substrate accumulating bacteria, if anaerobic conditions are followed by aerobic conditions could act as carbon source for denitrification in post-denitrification system. First exploratory batch tests, where the glycogen evolution was monitored, corroborate this assumption. PMID- 16045966 TI - Effects, transfer, and fate of RDX from aged soil in plants and worms. AB - The objectives of this study were to provide data that can be used to predict exposure-based effects of RDX in aged soil on multiple endpoint organisms representing two trophic levels. These data can be used for defining criteria or reference values for environmental management and conducting specific risk assessment. Dose-response experiments formed the basis for the evaluation of toxic effects and transfer of contaminants from soil into two trophic levels. Long-term exposure tests were conducted to evaluate chronic, sublethal, toxicity and transfer of aged soil-based explosives, with RDX as main contaminant. In these tests, plants were exposed for 55 days in the greenhouse, biomass was determined and residues of explosives parent compounds and RDX metabolites were analyzed using HPLC techniques. Worms were exposed for 28 days (Eisenia fetida) and 42 days (Enchytraeus crypticus) in the laboratory, biomass and number were determined, and tissues were analyzed for explosives compounds. The plants tolerated concentrations up to 1,540 mg RDX kg(-1) soil-DW. Biomass of Lolium perenne was not significantly related to soil-RDX concentration, while biomass of Medicago sativa significantly increased. No screening benchmark for RDX in soil for plants was calculated, since concentrations up to 1,540 mg kg(-1) soil failed to reduce biomass by 20% as required for a LOEC. RDX, RDX-metabolite MNX, and accompanying HMX concentrations in plants were significantly related to concentrations in soil after 55 days of exposure (RDX: R(2) = 0.77-0.89; MNX R(2) = 0.53-0.77; HMX: R(2) = 0.67-0.71). The average bioconcentration factors (BCF) were for RDX 17 in L. perenne and 37 in M. sativa, and for HMX 2 in L. perenne and 44 in M. sativa. The worms also tolerated concentrations up to 1,540 mg RDX kg(-1) soil-DW. Biomass of E. fetida adults decreased with soil-RDX concentration, and a LOEC of 1,253 mg kg(-1) soil-DW was estimated. RDX concentrations in E. fetida were significantly related to concentrations in soil after 28-day exposure (R(2) = 0.88). The average BCF in E. fetida for RDX was 1. Because in response to exposure to RDX-contaminated soil the RDX concentrations in plants increased initially and decreased subsequently, while those in worms increased continuously, RDX in worm tissues may accumulate to higher concentrations than in plant tissues, regardless of the low average BCF for worms. PMID- 16045967 TI - Developmental and behavioral effects of embryonic exposure to the polybrominated diphenylether mixture DE-71 in the killifish (Fundulus heteroclitus). AB - Exposures to penta polybrominated diphenylether (PeBDE) cause neurobehavioral toxicity in developing mice and rats. As levels of these ubiquitous contaminants are increasing in the environment, this raises concern that wildlife may also suffer such effects, with consequences for their ability to catch prey and avoid predators. PeBDE levels in wild-caught fish have been steadily escalating over the past fifteen years. To our knowledge, behavioral consequences of piscine embryonic exposure to PeBDE has not yet been studied. The objectives of this investigation were to characterize effects on development in an environmentally relevant fish model, and test for latent behavioral effects following cessation of exposure. Embryos from the estuarine minnow, Fundulus heteroclitus, were exposed from day 0-7 post fertilization to the industrial PeBDE mixture, DE-71 (0.001 to 100 microg l(-1)). Embryos were assayed for hatching success, development, and microsomal enzyme cytochrome P4501A (CYP1A) activity, which was determined by analysis of in ovo ethoxy-resorufin-O-deethylase (EROD) activation in embryos. Larval fish were assayed for predation ability, activity level, and fright response to a simulated predator. Juvenile fish were assayed for learning ability in a three-chambered fish maze. No induction of embryonic EROD activity was observed, nor was a high dose of DE-71 able to inhibit EROD activity induced by beta-naphthoflavone. No deformities were detected, but a subtle developmental asymmetry with respect to tail curvature direction was observed, and a hatching delay of up to 4.5 days was noted. Behavioral test results suggest that embryonic exposure to DE-71 may alter activity level, fright response, predation rates, and learning ability in subsequent life stages. PMID- 16045968 TI - Evaluation of biodegradation kinetic constants for aromatic compounds by means of aerobic batch experiments. AB - Kinetics of aerobic biodegradation have been investigated for twenty aromatic species using sludges collected from the aeration basin of municipal sewage treatment plants. The reproducibility of the results is tested with respect to the sludges period of collection and the wastewater treatment plant where they are taken. The comparison of kinetic constants, estimated for the investigated chemicals, allows to evaluate the reactivity effect of single groups (i.e., -OH, CH3, -Cl, -NO2) into the aromatic structures. The search for easy structure reactivity relationships is also attempted by means of contributing group methods. PMID- 16045969 TI - The palaeoecology of the micromammals from the late middle Pleistocene site of Hoedjiespunt 1 (Cape Province, South Africa). AB - The palaeontological site of Hoedjiespunt 1 (HDP1) represents a fossilized hyaena lair. A rich mammalian fauna, including four hominid teeth, has been recovered from the site. Micromammals were recovered from the same sediments as the larger fauna. Taphonomic analysis suggests that the micromammal assemblages from HDP1 were accumulated by a barn owl. The barn owl produces micromammal assemblages that provide a broad sample of micromammals, within a certain size range, living in the hunting area of the owl. There are size-related and other biases inherent in the prey selection of this predator, and owls may roost in one area and hunt in another however, the barn owl has frequently been found to provide a better indication of micromammals living within an area than trapping. The micromammals from HDP1 were used to reconstruct the microhabitats in the vicinity of the site. Two taxonomic habitat indexes were used to assess the environment and dominant habitat types at Hoedjiespunt 1. The variability and adaptability of many of the southern African micromammals complicates interpretation of the results, however, it appears that the micromammals from the HDP1 fossil assemblages utilized habitats of open, scrub vegetation, and rocky and sandy areas. It is suggested that the environment was not markedly different from today, but it may have been relatively more arid. A comparison between HDP1 and other fossil sites in the area dating from the terminal Pleistocene to the Holocene indicates that HDP1 is lacking certain species that are common to all the other west coast fossil sites. There is some discrepancy in the environment indicated by the large mammals as compared that indicated by to the micromammals at the site. It is suggested that this discrepancy may reflect the fact that an owl is likely to have hunted in the vicinity of the hyaena den, probably in the more open areas around the roost site, whereas the macrofauna, accumulated by the further-ranging brown hyaena (Hyaena brunnea), represents environments from further afield. PMID- 16045970 TI - Ro5-4864, a synthetic ligand of peripheral benzodiazepine receptor, reduces aging associated myelin degeneration in the sciatic nerve of male rats. AB - The peripheral-type benzodiazepine receptor (PBR) is a protein predominantly located in the mitochondrial outer membrane that plays an important role in the regulation of cell survival and proliferation. Previous studies have shown an enhanced expression of PBR in the regenerating sciatic nerve, suggesting that this protein may be involved in the regenerative response. The rat sciatic nerve suffers important structural alterations with aging, including alterations in the morphology of myelin sheaths and a decrease in the number of myelinated fibers. In this study, we have assessed the effect of two PBR ligands, Ro5-4864 and PK 11195, to determine whether PBR may influence aging-associated morphological changes in the sciatic nerve. The treatment of 23-month-old, Sprague-Dawley male rats for 1 month with Ro5-4864 significantly reduced the percentage of fibers with myelin decompaction and increased the total number of myelinated fibers. In contrast, PK-11195, a PBR ligand that binds to a different site than Ro5-4864 in the PBR molecule, did not significantly affect any of the parameters analyzed. These findings support the potential role of PBR ligands to prevent aging associated peripheral nerve degeneration. PMID- 16045971 TI - Novel mechanisms of aspirin pharmacologic actions: a model for studying herbal natural products. PMID- 16045972 TI - Effects of donepezil, nicotine and haloperidol on the central serotonergic system in mice: implications for Tourette's syndrome. AB - We have previously reported that acute and chronic donepezil and nicotine administration significantly attenuate DOI-induced head twitch response (HTR) in mice. This behavior, primarily mediated by stimulation of 5-HT2A receptors, has been proposed to model tic symptoms seen in Tourette's syndrome (TS). Haloperidol, a drug widely used to treat symptoms of TS, has also been reported to reduce DOI-induced head shakes in rodents when administered acutely. These findings suggest an inhibitory interaction of these drugs with 5-HT2A receptors. To test this hypothesis, we evaluated the effects of chronic donepezil, nicotine and haloperidol on expression levels of 5-HT2A mRNA and 5-HT2A receptor density in select brain regions. Initially, we established a dose-response relationship for the acute and chronic haloperidol and DOI-induced HTR. Male ICR mice were treated twice daily with donepezil (0.1 mg/kg), nicotine (0.5 mg/kg), and once daily with haloperidol (0.4 mg/kg) for 14 days and were sacrificed 16-18 h after the last injection. These drug regimens were chosen because of their significant effects on DOI-induced HTR. Donepezil significantly increased 5-HT2A mRNA level, but not the receptor density in the striatum. In the midbrain, donepezil significantly decreased the receptor density without affecting the 5-HT2A mRNA level. In the frontal cortex, only haloperidol significantly reduced the 5-HT2A receptor density. The cortex was the only area where donepezil, nicotine and haloperidol significantly reduced the 5-HT2A receptor density. The results suggest that the anti-tic properties of donepezil, nicotine and haloperidol in this paradigm might be due to antagonism of cortical 5-HT2A receptors. Thus, further investigation of involvement of cortical 5-HT2A receptors in TS as well as evaluation of selective 5-HT2A receptor antagonists in this disorder is warranted. PMID- 16045973 TI - Application of sensitive enzymeimmunoassays for oxytocin and prolactin determination in blood plasma of yaks (Poephagus grunniens L.) during milk let down and cyclicity. AB - Highly sensitive and specific enzymeimmunoassays for oxytocin and prolactin determination in yak plasma using the biotin-streptavidin amplification system and the second antibody coating technique were validated and applied for determining their profiles during milk let down and cyclicity in yaks. Oxytocin EIA was conducted taking duplicate 200 microl of unknown plasma samples and standards per well. The lowest detection limit was 0.2 pg/well, which corresponded to 1pg/ml plasma. Prolactin EIA was carried out directly in 50 microl of yak plasma. The sensitivity of EIA procedure was 5 pg/well prolactin, which corresponded to 0.1 ng/ml plasma. Mean plasma prolactin concentrations although high at estrus were not statistically different (P > 0.05) from the hormone concentrations on other days. Mean plasma prolactin concentrations during non-breeding season were significantly higher (P < 0.001) than that recorded in breeding season. Oxytocin and prolactin profiles were also obtained in two yaks before, during and after milking. A sharp release of oxytocin and prolactin shortly after udder stimulation was observed. High levels of oxytocin and prolactin were maintained during milking, falling sharply thereafter. PMID- 16045974 TI - The variability of ovum pick-up response and in vitro embryo production from monozygotic twin cows. AB - Oocytes were recovered by ovum pick up (OPU) from nine pairs of monozygotic twin German Simmental cows. The hypothesis was that there is less variability between identical twins versus among non-related individuals in the variation in the recovery of oocytes by OPU and in the efficiency of in vitro embryo production. Estrous cycles were synchronized with two doses of cloprostenol, 11 days apart. Beginning 3-4 days after synchronized estrus, OPU was done twice weekly (every 3 or 4 days; total of 11 sessions). The influence of repeated OPU on estrous cyclicity was established by estrus detection, plasma progesterone concentrations, and ovarian ultrasonography. There were no differences among days of collection for the number and quality of cumulus oocyte-complexes (COCs), and rates of cleavage and blastocyst formation. A total of 1,661 COCs, including 657 (39.6%) good-quality COCs, were recovered. From 1,457 (87.7%) cultured COCs, 827 zygotes cleaved and 314 blastocysts were produced on Day 7. The total number of COCs and the blastocyst rates varied among pairs of monozygotic twins; within pairs, only slight differences were observed. In conclusion, recovery of COCs and production of embryos had substantially less variation within pairs of monozygotic twins than among non-related cattle. PMID- 16045975 TI - The cell agglutination agent, phytohemagglutinin-L, improves the efficiency of somatic nuclear transfer cloning in cattle (Bos taurus). AB - One of the several factors that contribute to the low efficiency of mammalian somatic cloning is poor fusion between the small somatic donor cell and the large recipient oocyte. This study was designed to test phytohemagglutinin (PHA) agglutination activity on fusion rate, and subsequent developmental potential of cloned bovine embryos. The toxicity of PHA was established by examining its effects on the development of parthenogenetic bovine oocytes treated with different doses (Experiment 1), and for different durations (Experiment 2). The effective dose and duration of PHA treatment (150 microg/mL, 20 min incubation) was selected and used to compare membrane fusion efficiency and embryo development following somatic cell nuclear transfer (Experiment 3). Cloning with somatic donor fibroblasts versus cumulus cells was also compared, both with and without PHA treatment (150 microg/mL, 20 min). Fusion rate of nuclear donor fibroblasts, after phytohemagglutinin treatment, was increased from 33 to 61% (P < 0.05), and from 59 to 88% (P < 0.05) with cumulus cell nuclear donors. The nuclear transfer (NT) efficiency per oocyte used was improved following PHA treatment, for both fibroblast (13% versus 22%) as well as cumulus cells (17% versus 34%; P < 0.05). The cloned embryos, both with and without PHA treatment, were subjected to vitrification and embryo transfer testing, and resulted in similar survival (approximately 90% hatching) and pregnancy rates (17-25%). Three calves were born following vitrification and embryo transfer of these embryos; two from the PHA-treated group, and one from non-PHA control group. We concluded that PHA treatment significantly improved the fusion efficiency of somatic NT in cattle, and therefore, increased the development of cloned blastocysts. Furthermore, within a determined range of dose and duration, PHA had no detrimental effect on embryo survival post-vitrification, nor on pregnancy or calving rates following embryo transfer. PMID- 16045976 TI - Final part-word repetitions in school-age children: two case studies. AB - In contrast to the many published accounts of the disfluent repetition of sounds at the beginnings of words, cases where it is predominantly the final parts of words that are repeated have been reported relatively rarely. With few exceptions, those studies that have been published have described either pre school children or neurologically impaired subjects. The purpose of this case report was to describe final part-word repetitions in the speech of two school age boys of normal intelligence with no known neurological lesions. Their speech was recorded during spontaneous conversation, reading, and sentence repetition. The repetitions occurred in all three speaking conditions, although the majority of instances were observed in spontaneous speech, and on both content words and function words. The participants exhibited no apparent awareness of the disfluencies, no abnormal muscle tension, and no accessory behaviours. Each child produced word-final repeated fragments whose phonological structure was highly predictable according to his individual set of rules. The results are discussed in terms of possible motor and cognitive explanations for the disfluencies. EDUCATIONAL OBJECTIVES: As a result of this activity, the participant will be able to: (1) summarize prior research into final part-word repetition; (2) describe the detailed characteristics of final part-word repetitions as displayed by two children of normal intelligence; and (3) discuss ways in which the behaviour might be explained as part of a model of speech production. PMID- 16045977 TI - Childhood stuttering and dissociations across linguistic domains. AB - The purpose of this investigation was to evaluate the possible presence of dissociations in the speech and language skills of young children who do (CWS) and do not stutter (CWNS) using a correlation-based statistical procedure [Bates, E., Appelbaum, M., Salcedo, J., Saygin, A. P., & Pizzamiglio, L. (2003). Quantifying dissociations in neuropsychological research. Journal of Clinical and Experimental Neuropsychology, 25, 1128-1153]. Participants were 45 preschool CWS and 45 CWNS between the ages of 3;0 and 5;11 (years;months), with the two groups matched by age, gender, race, and parental socioeconomic status. Children participated in a parent-child interaction for the purpose of disfluency analysis and responded to four standardized speech-language tests for subsequent analyses as main dependent variables. Findings indicated that CWS were over three times more likely than CWNS to exhibit dissociations across speech-language domains, with 44 cases of dissociation for CWS and 14 for CWNS across 10 possible comparisons. Results suggest that there may be a subgroup of CWS who exhibit dissociations across speech-language domains, which may result in a greater susceptibility to breakdowns in speech fluency. EDUCATIONAL OBJECTIVES: The reader will be able to: (1) summarize findings from previous studies examining differences in speech and language performance between children who do and do not stutter; (2) describe what is meant by "dissociations" in the speech and language skills of young children who do and do not stutter; and (3) discuss three hypotheses that could account for the present findings that suggest CWS, more often than CWNS, exhibit dissociations in their speech-language system. PMID- 16045978 TI - The effects of boric acid and phosphoric acid on the compressive strength of glass-ionomer cements. AB - OBJECTIVES: Both boric acid (H3BO3) and phosphoric acid (H3PO4) are components of dental cements, commonly incorporated into glass (as ingredients in the melt) and occasionally added to the powder or liquid components. This study investigated the effect of boric acid addition to an experimental glass-ionomer powder and the effect of phosphoric acid addition to a glass-ionomer liquid on the 24-h compressive strength. METHODS: Boric acid powder was added in various concentrations to an experimental glass-ionomer powder and, separately, phosphoric acid was added to an experimental glass-ionomer liquid. Powders and liquids were dosed into capsules at various powder:liquid ratios and cements thus formed were assessed for 24-h compressive strength. RESULTS: Incorporation of boric acid in glass-ionomer powder resulted in a pronounced decrease (p < 0.05 at 1% boric acid) in compressive strength. Addition of phosphoric acid produced initially stronger cements (up to 13% increase at 1% phosphoric acid) before also declining. SIGNIFICANCE: The incorporation of less than 2% w/w phosphoric acid in glass-ionomer liquids may improve cement strengths without compromising clinical usefulness. The incorporation of boric acid in glass-ionomer cements is contraindicated. PMID- 16045979 TI - Comparative assessment of the roughness, hardness, and wear resistance of aesthetic bracket materials. AB - OBJECTIVE: The purpose of this study was to assess critical properties of orthodontic aesthetic bracket materials. METHOD: Samples of polycarbonate, poly(oxy)methylene, ultra high molecular weight polyethylene (UHMWPE), and polycrystalline alumina raw material used from bracket manufacturing were subjected to: (a) profilometry for the determination of Ra, Rq, Rmax and Rz roughness parameters; (b) Vickers hardness (HV50) testing; and (c) wear resistance determined by the scratch test. The results were analyzed with one-way ANOVA and Tukey multiple comparisons test at alpha=0.05 level of significance. RESULTS: The UHMWP and alumina specimens showed the highest roughness values for all parameters. The highest hardness, amongst the polymeric raw materials, was obtained from the poly(oxy)methylene appliances. Differences were also noted between the polycarbonate raw material of different manufacturers implying an effect from the manufacturing process. With the exception of alumina, the highest wear resistance was found for the poly(oxy)methylene specimens. SIGNIFICANCE: The results of this study reveal the variability among aesthetic plastic bracket raw materials, their reduced hardness and wear resistance relative to alumina as well as the inappropriateness of UHMWPE as alternative bracket material. PMID- 16045980 TI - Polymerization characteristics of light- and auto-curing resins for individual splints. AB - OBJECTIVES: The aim of this in vitro study was to assess different resins for occlusal splints regarding a number of polymerization characteristics. METHODS: One autopolymerizing resin (Palapress) and four light-curing resins (Acrylight, Primosplint, Triad TranSheet Colorless and Pink) were tested for flexural strength, modulus of elasticity, water sorption, volumetric shrinkage, and conversion of double bonds. These resins were polymerized for 10 and 15 min in a light oven at frequencies between 320 and 400 nm or 400 and 500 nm, respectively. Data were treated statistically by ANOVA and by Scheffe test. RESULTS: Concerning flexural strength and modulus of elasticity, Triad TranSheet Colorless showed significantly greater values than Primosplint and Palapress (p<0.05). The light curing resins Acrylight and Primosplint featured significantly higher water sorption when cured for 10 min compared to all other resins (p<0.05). Acrylight displayed significantly higher volumetric shrinkage and a higher quantity of remaining double bonds compared to all other light-curing resins (p<0.05). Primosplint, Triad TranSheet Colorless and Pink showed significantly lower shrinkage values than autopolymerizing Palapress (p<0.05). For all light curing resins, polishing of the samples resulted in a significantly lower quantity of remaining double bonds (p<0.05) compared to unpolished samples. SIGNIFICANCE: With respect to the polymerization properties evaluated, the light-curing resins reached similar material properties to the autopolymerizing resin or even surpassed them. Light-curing resins seem to be suitable for the fabrication of occlusal splints and can been seen as an alternative to autopolymerizing polymethylmethacrylates. PMID- 16045981 TI - A preliminary study of the interaction of glass-ionomer dental cements with amino acids. AB - OBJECTIVES: The aim of the study was to determine how the presence of low concentrations of the amino acids glycine and alanine affected the interaction of restorative glass-ionomer cements with water. METHODS: For each material (Fuji IX and Ketac Molar, both handmixed), sets of six cylindrical specimens of 4 mm diameter x 6 mm height were prepared and weighed; then stored individually in one of: deionized water, 0.1% by mass glycine, 0.5% glycine, 0.1% alanine (Sigma Aldrich) and 0.5% alanine. After 1 week, the specimens were re-weighed and the pH of each solution determined. The solutions were then bulked and analyzed for ion content (Na, Ca, Al, Si and P) using ICP-OES. In a separate series of experiments, disks (12 mm diameter x 1 mm thick) were prepared, and exposed to similar solutions for periods of a week, after which Vicker's hardness was determined. Differences were analysed by one-way ANOVA, followed by Neumann-Keuls post-hoc analysis (P<0.05). RESULTS: All specimens increased in weight, and altered pH of the storage solution to similar values. Both cements released less aluminum, but generally more sodium, calcium and silicon in the presence of amino acids. Little or no phosphorus was found to be released under any conditions. Surface hardness of Fuji IX was not influenced by the presence of amino acids, but for Ketac Molar it was generally lower in the presence of amino acids. SIGNIFICANCE: These results show that the interaction of glass-ionomer cements with water is influenced by the presence of low concentrations of amino acids, and that this may alter surface hardness. PMID- 16045982 TI - Ligament tissue engineering: an evolutionary materials science approach. AB - The anterior cruciate ligament (ACL) is important for knee stabilization. Unfortunately, it is also the most commonly injured intra-articular ligament. Due to poor vascularization, the ACL has inferior healing capability and is usually replaced after significant damage has occurred. Currently available replacements have a host of limitations, this has prompted the search for tissue-engineered solutions for ACL repair. Presently investigated scaffolds range from twisted fiber architectures composed of silk fibers to complex three-dimensional braided structures composed of poly (L-lactic acid) fibers. The purpose of these tissue engineered constructs is to apply approaches such as the use of porous scaffolds, use of cells, and the application of growth factors to promote ligament tissue regeneration while providing mechanical properties similar to natural ligament. PMID- 16045983 TI - Improved synthesis and in vitro characterization of chitosan-thioethylamidine conjugate. AB - The aim of this study was to establish improved reaction conditions for the synthesis of chitosan-thioethylamidine (Ch-TEA) conjugate and to evaluate the properties of the obtained Ch-TEA conjugate. The influence of different factors on the coupling reaction, such as concentration of chitosan solution, employment of reducing agent and deprotection of S-acetyl groups, was evaluated. The cohesive properties and stability of the obtained conjugate were evaluated by disintegration test and by oxidation experiments, respectively. The adhesive properties of Ch-TEA conjugate were evaluated in vitro on freshly excised porcine mucosa via tensile studies and the rotating cylinder method. The permeation enhancing effect of Ch-TEA conjugate was evaluated in Ussing-type chambers by using rhodamine 123 as model compound. The resulting conjugate displayed 225 micromol immobilized free thiol groups and 102 micromol disulfide bonds per gram polymer. The degree of modification depends mostly on the chitosan concentration employed and the deprotection of S-acetyl groups with hydroxylamine. During oxidation studies the amount of thiol groups decreased by 61%. Disintegration studies of tablets comprising Ch-TEA demonstrated stability for 48 h. In tensile studies, the total work of adhesion of the conjugate was determined to be 5.1 fold increased in comparison to unmodified chitosan. Results from the rotating cylinder method showed more than a 13-fold increase in the adhesion time of thiolated chitosan versus unmodified chitosan. The apparent permeability coefficient (Papp) of the system 0.5% (w/v) Ch-TEA conjugate with 5% (w/v) glutathione was calculated to be 5.35 x 10(-8)cm/s, while the Papp value of the system 0.5% (w/v) unmodified chitosan was 1.73 x 10(-8)cm/s. These features should render Ch-TEA useful as an excipient for various drug delivery systems. PMID- 16045984 TI - Ligand density characterization of peptide-modified biomaterials. AB - A simple fluorescence based characterization method was developed to assess ligand density on peptide-modified biomaterials. The method exploits the exquisite sensitivity of proteolysis for the purpose of liberating a fluorescently labeled probe fragment from an immobilized peptide. The released fragment can then be detected in solution using high-throughput fluorometry. In silico screening tools identified the enzyme chymotrypsin as a promising candidate for releasing a detectable probe fragment from the fluorescently labeled peptide, Ac-CGGNGEPRGDTYRAYK(FITC)GG-NH(2). After chymotrypsin digestion of the peptide in solution was first characterized using mass spectrometry and HPLC, a basic enzyme mediated release protocol was developed and implemented to generate peptide-binding isotherms on various peptide-modified biomaterials. The new method is sensitive, has good signal-to-noise ratio (S/N), and is easily standardized. Furthermore, the technique can be applied independent of material chemistry and geometry, making it a suitable alternative to radiolabeling for a wide range of biomaterial applications. PMID- 16045985 TI - Molecular polymorphism and expression analysis of MHC class II B gene from red sea bream (Chrysophrys major). AB - MHC class II (major histocompatibility complex class II) plays an important role in the immune response of vertebrates. Its function is to present antigenic peptides to the T-cell receptor. In order to study the function and molecular polymorphism of class II B gene in fish, we have isolated cDNAs encoding class II B from spleen cDNA library of red sea bream (Chrysophrys major) by using EST sequencing, and examined genomic organization, molecular polymorphism and expression of red sea bream class II B gene. As in other vertebrates, five exons and four introns were identified in red sea bream class II B gene. Seven class II B alleles were identified from seven individuals of red sea bream. The deduced amino acid sequence of red sea bream MHC class II B 1(Chma-DAB*0101) had 87.1, 85.1, 87.1, 90.4, 87.1, 90.8% identity with those of red sea bream class II B 2, 3, 4, 5, 6, 7(Chma-DAB*0201-Chma-DAB*0701), respectively, and had 75.2, 74.5, 55.9, 55.1, 34.3 and 30.4% identity with those of striped sea bass, cichlid, rainbow trout, Atlantic salmon, mouse and human, respectively. Four different class II B alleles were observed in a single individual and two different 3' untranslated region (3' UTR) sequences from this individual may infer the existence of two loci at least. Semi-quantitative RT-PCR demonstrated that high expression was detected in liver, head kidney, kidney, intestine, gill, stomach, hear and spleen, low expression in muscle and blood. Challenge of red sea bream with the pathogenic bacteria, Vibrio anguillarum, resulted in a significant decrease in the expression of MHC class II B mRNA from 5 to 72 h after infection in liver, spleen, head kidney and intestine, followed by a recovery to normal level after 96 h. PMID- 16045986 TI - Health effects in children aged 3-6 years induced by environmental lead exposure. AB - OBJECTIVES: To investigate the involvement of oxidative damage in lead-induced toxicity in children aged 3-6 years and to enlighten whether oxidative stress indicators are correlated with the known indices of lead toxicity. METHODS: Blood samples were collected from 408 subjects (217 boys and 191 girls) in the urban kindergartens. The age range of the subjects was 3-6 years. Blood lead levels (BLLs) were analyzed by flameless atomic absorption spectrophotometry. Activities of delta-aminolevulinic acid dehydratase (ALAD), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and contents of glutathione (GSH) in erythrocyte and levels of plasma malondialdehyde (MDA) were analyzed spectrophotometrically in these children. RESULTS: Children with BLLs >or = 100 microg/L had significantly decreased erythrocyte ALAD activities and increased plasma MDA levels compared to the children with BLLs < 100 microg/L. No significant changes were observed in erythrocyte SOD and GSH-Px activities and GSH levels associated with elevated BLLs in these children. CONCLUSION: Present data indicate that oxidative damage could be induced by lead in children with BLLs > or = 100 microg/L, and this may partly be attributed to the inhibited ALAD activities. Statistically significant changes of oxidative stress parameters in preschool children while BLLs were more than 100 microg/L could be implicated that oxidative damage might contribute to lead-induced intellectual impairment. PMID- 16045987 TI - The role of dopamine in conditioning and latent inhibition: what, when, where and how? AB - It is well established that dopamine is released in the nucleus accumbens (NAC) in animals in rewarding or reinforcing situations, and widely believed that this release is the substrate of, or at least closely related to, the experience of reward. The demonstration of conditioned release of dopamine by stimuli conditioned to primary rewards has reinforced this view. However, a number of observations do not sit comfortably with this interpretation, most notably that dopamine is released equally effectively in NAC by aversive stimuli, and stimuli conditioned to them. Furthermore, additional release of dopamine is seen during conditioning, even if motivational stimuli of either type are not involved. It is suggested here that one important action of NAC dopamine release is to restore the salience of potential conditioned stimuli, when this has been reduced by prior un-reinforced experience. The paradigm of latent inhibition (LI) demonstrates a behavioural effect of this type, and extensive studies on the role of dopamine in LI have been undertaken by us and others. Those studies are reviewed here, together with some previously unpublished data, to demonstrate that (1) amphetamine disruption of LI is indeed a function of calcium-dependant dopamine release in the NAC at the time of conditioning; (2) other drugs acting on LI via changes in dopamine transmission act at the same locus; (3) the disruptive effect of indirect dopamine agonists on LI can be prevented by either D-1 selective receptor antagonists, or D-2 selective receptor antagonists. It is concluded that dopamine release in these very varied behavioural contexts (reward, punishment, conditioning, modulation of salience) must be differentiated in some way, and that this should be investigated. An alternative explanation, if they are not differentiated, would be that the release in fact does have the same functional significance in each case. We suggest that this common significance might be the broadening of attention to take in potentially conditionable stimuli, which have previously been devalued. PMID- 16045988 TI - Structural and functional characterization of glycosylation in an immunoglobulin G1 to Cryptococcus neoformans glucuronoxylomannan. AB - Analysis of the N-linked oligosaccharides of the murine IgG1 monoclonal antibody (mAb) to Cryptococcus neoformans by LC/MS revealed five different core fucosylated, biantennary complex-type oligosaccharides at Asn-293, with the major species being a mono-galactosylated oligosaccharide with the glycosyl composition of Hex4HexNAc4Fuc (39% of the total glycan pool). The primary sequence predicted from nucleic acid sequencing differed from that measured by mass spectrometry at position 33 (ASN to ASP), a finding that may represent post-translational modification caused by spontaneous ASP deamination. Analysis of mAb 18B7 from three hybridoma clones revealed the same heterogenous N-glycan pattern, indicating that diversity in oligosaccharide structures originated from individual cells. The binding of native and de-glycosylated mAb 18B7 to cryptococcal Ag was comparable but the de-glycosylated 18B7 had shorter serum half-life and did not activate complement (C). De-glycosylated mAb 18B7 was opsonic for C. neoformans with murine macrophages through a mechanism that involved C-independent ingestion through the C receptor. Passive administration of de-glycosylated mAb 18B7 mediated comparable protective efficacy to the native mAb in mice with lethal infection. The results imply that the contribution of N glycan structure to immunoglobulin function varies depending on the Ag-Ab system. PMID- 16045989 TI - Gene expression profiling of the effects of intravenous immunoglobulin in human whole blood. AB - Intravenous immunoglobulin (IVIG) is involved in many complex mechanisms that act in synergy including expression and function of Fc receptors, complement activation, the cytokine network, interaction with the anti-idiotypic network and modulation of B and T cell activation. To gain insight into the early effects of IVIG on this broad range of activities at the gene level we performed DNA microarray analysis. Human whole blood was incubated in vitro for 4 h followed by extraction of RNA which was hybridized to a chip containing 8793 genes. About 75 upregulated genes and 21 downregulated genes were identified using a cut off for the false discovery rate of 5%. These genes are associated with a wide range of cellular immune functions in line with the broad mechanism of action of IVIG. A striking upregulation of a series of genes coding for chemokines was measured. This finding was confirmed at the protein level as pharmacologically relevant concentrations of CXCL9 and CXCL10 were measured in serum. Interestingly, IVIG shows a partial overlap of its gene expression program with lipopolysaccharide. Our data suggests multiple hypotheses regarding the pharmacology of IVIG that must be validated by complementary studies. PMID- 16045990 TI - A unique Vgamma5-T-cell population in the murine mammary gland. AB - Mammary epithelia and the epidermis share a common embryologic origin. Like resident epidermal T cells (ETC), a significant number of murine mammary gland T cells (MTC) express Vgamma5-T-cell receptors (TCR). MTC were assessed to see if they express the same invariant Vgamma5/Vdelta1-TCRs as those of ETC. MTC-derived hybridomas, positive for Vgamma5-TCRs, expressed Vdelta5 instead of Vdelta1. Additionally, they showed non-germline additions in the gamma-TCR and delta-TCR junctions, sharing among them an identical 19-nucleotide N-region in the delta TCR junctions. Vgamma5-TCR and Vdelta5-TCR gene sequences from fresh peripartum MTC also contained variable N-regions, including a 7-nucleotide N-region similar to one found in the hybridomas. Thus, Vgamma5-TCR+ MTC are distinct from Vgamma5 TCR+ ETC by expressing variable non-canonical TCRs, predicting differences in the two T-cell populations in ligand specificity and in function. PMID- 16045991 TI - Aluminium, antiperspirants and breast cancer. AB - Aluminium salts are used as the active antiperspirant agent in underarm cosmetics, but the effects of widespread, long term and increasing use remain unknown, especially in relation to the breast, which is a local area of application. Clinical studies showing a disproportionately high incidence of breast cancer in the upper outer quadrant of the breast together with reports of genomic instability in outer quadrants of the breast provide supporting evidence for a role for locally applied cosmetic chemicals in the development of breast cancer. Aluminium is known to have a genotoxic profile, capable of causing both DNA alterations and epigenetic effects, and this would be consistent with a potential role in breast cancer if such effects occurred in breast cells. Oestrogen is a well established influence in breast cancer and its action, dependent on intracellular receptors which function as ligand-activated zinc finger transcription factors, suggests one possible point of interference from aluminium. Results reported here demonstrate that aluminium in the form of aluminium chloride or aluminium chlorhydrate can interfere with the function of oestrogen receptors of MCF7 human breast cancer cells both in terms of ligand binding and in terms of oestrogen-regulated reporter gene expression. This adds aluminium to the increasing list of metals capable of interfering with oestrogen action and termed metalloestrogens. Further studies are now needed to identify the molecular basis of this action, the longer term effects of aluminium exposure and whether aluminium can cause aberrations to other signalling pathways in breast cells. Given the wide exposure of the human population to antiperspirants, it will be important to establish dermal absorption in the local area of the breast and whether long term low level absorption could play a role in the increasing incidence of breast cancer. PMID- 16045992 TI - The efficacy of deferiprone on tissues aluminum removal and copper, zinc, manganese level in rabbits. AB - The effect of 1,2-dimethyl-3-hydroxypyrid-4-one [deferiprone (DE)] on aluminum mobilization and elimination from tissues and serum as well as the influence on the excretion of trace elements, copper, zinc and manganese in rabbits was investigated. Sixteen New Zealand rabbits were randomly divided into three groups: control, Al-only and Al+DE. The Al-only and Al+DE animals received injections of Al2(SO43.18H2O 600 micromol Al/kg 5 days per week for 3 weeks. One week after the last Al injection the Al+DE rabbits were given deferiprone 750 micromol/kg/day intragastrically for 2 weeks. At the 42nd day the animals were sacrificed and the organs were taken and digested. Blood was taken from the ear artery three times (at the initiation of the experiment, before and after deferiprone administration). The aluminum and copper, zinc, manganese were determined by atomic absorption spectrophotometry. Our results showed that deferiprone could highly mobilize aluminum stores from tissues. At the end of experiment the aluminum contents of bone, kidney, liver and brain in Al+DE were significantly lower than that in Al-only rabbits. The copper, zinc, manganese contents were not affected by deferiprone administration. PMID- 16045993 TI - RAPD for the typing of coagulase-negative staphylococci implicated in catheter related bloodstream infection. AB - OBJECTIVES: A rapid random amplification of polymorphic DNA (RAPD) technique was developed to distinguish between strains of coagulase-negative staphylococci (CoNS) involved in central venous catheter (CVC)-related bloodstream infection. Its performance was compared with that of pulsed-field gel electrophoresis (PFGE). METHODS: Patients at the University Hospital Birmingham NHS Foundation Trust, U.K. who underwent stem cell transplantation and were diagnosed with CVC related bloodstream infection due to CoNS whilst on the bone marrow transplant unit were studied. Isolates of CoNS were genotyped by PFGE and RAPD, the latter employing a single primer and a simple DNA extraction method. RESULTS: Both RAPD and PFGE were highly discriminatory (Simpson's diversity index, 0.96 and 0.99, respectively). Within the 49 isolates obtained from blood cultures of 33 patients, 20 distinct strains were identified by PFGE and 25 by RAPD. Of the 25 strains identified by RAPD, nine clusters of CoNS contained isolates from multiple patients, suggesting limited nosocomial spread. However, there was no significant association between time of inpatient stay and infection due to any particular strain. CONCLUSION: The RAPD technique presented allows CoNS strains to be genotyped with high discrimination within 4h, facilitating real-time epidemiological investigations. In this study, no single strain of CoNS was associated with a significant number of CVC-related bloodstream infections. PMID- 16045994 TI - Rapid serodiagnosis of Staphylococcus aureus surgical site infection following median sternotomy. AB - OBJECTIVES: To determine the sensitivity and specificity of a novel ELISA for the serodiagnosis of surgical site infection (SSI) due to staphylococci following median sternotomy. METHODS: Twelve patients with a superficial sternal SSI and 19 with a deep sternal SSI due to Staphylococcus aureus were compared with 37 control patients who also underwent median sternotomy for cardiac surgery but exhibited no microbiological or clinical symptoms of infection. A further five patients with sternal SSI due to coagulase-negative (CoNS) staphylococci were studied. An ELISA incorporating a recently recognised exocellular short chain form of lipoteichoic acid (lipid S) recovered from CoNS, was used to determine serum levels of anti-lipid S IgG in all patient groups. RESULTS: Serum anti-lipid S IgG titres of patients with sternal SSI due to S. aureus were significantly higher than the control patients (P<0.0001). In addition, patients with deep sternal SSI had significantly higher serum anti-lipid S IgG titres than patients with superficial sternal SSI (P = 0.03). Serum anti-lipid S IgG titres of patients with sternal SSI due to CoNS were significantly higher than the control patients (P = 0.001). CONCLUSION: The lipid S ELISA may facilitate the diagnosis of sternal SSI due to S. aureus and could also be of value with infection due to CoNS. PMID- 16045995 TI - An unusual transitory increase of lupus anticoagulant in dengue virus infection complicated with cerebral ischaemia. AB - Dengue is a common mosquito-transmitted viral disease prevalent at many undeveloped and developing countries. Although, neurological complication and hemostatic disturbance are common in dengue virus infection, cerebral ischaemia is barely mentioned. A 61-year-old woman experienced an acute onset of right hemiparesis in her early course of dengue virus type II infection. Extensive laboratory investigation revealed an increase of lupus anticoagulant activity and a prolonged activated partial thromboplastin time that were normalized in the convalescent stage. Her human leucocyte antigens were A2, A11, B8, Cw7, DR4 and DR9. This patient highlights the co-operation between transient activation of humoral immunity and preferential immunogenetic for coagulopathic thrombosis in specific viral disease. PMID- 16045996 TI - Proposal for vaccination against SARS coronavirus using avian infectious bronchitis virus strain H from The Netherlands. PMID- 16045997 TI - Immunopharmacology--antibodies for specific modulation of proteins involved in neuronal function. AB - The application of antibodies to living neurones has the potential to modulate function of specific proteins by virtue of their high specificity. This specificity has proven effective in determining the involvement of many proteins in neuronal function where specific agonists and antagonists do not exist, e.g. ion channel subunits. We discuss studies where antibodies modulate functions of voltage gated sodium, voltage gated potassium, voltage gated calcium hyperpolarisation activated cyclic nucleotide (HCN gated) and transient receptor potential (TRP) channels. Ligand gated channels studied in this way include nicotinic acetylcholine receptors, purinoceptors and GABA receptors. Antibodies have also helped reveal the involvement of different intracellular proteins in neuronal functions including G-proteins as well as other proteins involved in trafficking, phosphoinositide signalling and neurotransmitter release. Some suggestions for control experiments are made to help validate the method. We conclude that antibodies can be extremely valuable in determining the functions of specific proteins in living neurones in neuroscience research. PMID- 16045998 TI - Macrophages and not granulocytes are involved in cervical ripening. AB - To clarify the role of leucocytes in human cervical ripening and dilatation, cervical biopsies were obtained from six non-pregnant women, eight women undergoing early termination of pregnancy, 18 pregnant women undergoing elective Caesarean section at term (both with and without a ripe cervix as determined by Bishop score) and 11 women after term vaginal delivery. Leucocytes were localised by immunohistochemistry labelling and quantified in subepithelial and deep stromal areas. CD45+ leucocytes were more numerous in the subepithelial area of the cervix than in the deep stroma in all groups (P<0.01). CD14+ macrophages and CD15+ granulocytes were increased in both the subepithelial and deep stromal areas only in the vaginal delivery group (P<0.01). The number of macrophages in the ripening cervix (Bishop score above 4) was higher than in the unripe cervix (Bishop score 4 or less; P<0.05) with no differences in other leucocyte populations. CD3+ CD8+ T cells in the subepithelial area were reduced in late pregnancy and after vaginal delivery (P<0.01), but showed no relationship to Bishop score. Macrophages and granulocytes may be involved in the process of cervical dilatation, but macrophage infiltration into the ripening cervix before labour suggests their role in the ripening process. Reduced numbers of CD3+ CD8+ T-lymphocytes in late pregnancy and after vaginal delivery suggests that local immunity is down-regulated in the late pregnancy period. Regional differences in leucocyte subpopulations in the cervix indicate that leucocyte infiltration is likely to be regulated by local factors. PMID- 16045999 TI - The transfer of maternal IgE and other immunoglobulins specific for Trichostrongylus colubriformis larval excretory/secretory product to the neonatal lamb. AB - The transference of immunoglobulins from six New Zealand Romney ewes to their lambs was examined. Immunoglobulin levels were determined in ewe plasma, colostrum and lamb plasma shortly after birth and before the lambs fed, in lamb plasma 2 days after birth, and lamb plasma, ewe plasma and milk 30 days after parturition. Levels of total IgE, and IgE, IgG1, IgG2, IgM, and IgA with specificity for Trichostronglus colubriformis third stage larval secretory/excretory products (TcL3E/S) were determined. Mean levels of total IgE were three times higher in colostrum than in parturient ewe plasma while only trace amounts were detected in milk at 30 days after birth (107.7, 34.3, and 0.2U ml(-1), respectively, differences between means P< or =0.01). Mean total IgE in lamb plasma rose from being undetectable before suckling to levels comparable to those of the ewes by 2 days after birth (21.7U ml(-1)) and then declined to low levels by 30 days (0.4U ml(-1)). Total IgE levels in lamb plasma were significantly correlated with levels in ewe plasma and colostrum (r=0.91, P< or =0.01; r=0.96, P< or =0.003, respectively). The transference of TcL3E/S-specific IgE, IgG1 and IgA was substantial with mean levels of these antibodies in lamb plasma at 2 days comparable to that in parturient ewe plasma (absorbance levels in lamb plasma of 0.283, 0.537, and 0.334, respectively). Proportionally less maternal IgM and IgG2 appeared to be transferred to the lambs (absorbance of 0.112 and 0.081, respectively). Levels of TcL3E/S-specific IgE and IgG1 in lamb plasma at 2 days were significantly correlated with levels in parturient ewe plasma and colostrum (r=0.89 and 0.82, 0.85 and 0.96; all P< or =0.05, respectively). These results indicate that IgE is concentrated in ewe colostrum and that substantial amounts of maternal IgE are transferred to lambs via colostrum. Further, the results suggest that humoral immunity against gastro intestinal nematode parasites and potentially other parasites in colostrum-fed lambs may approximate that of the ewe. The implications of the transference of humoral immunity through colostrum in ruminants for the passive protection and the development of active immunity against parasites remains to be fully explored. PMID- 16046000 TI - The BDNF-Val66Met polymorphism: implications for susceptibility to multiple sclerosis and severity of disease. AB - Neurodegeneration following inflammatory injury is considered to be a pathological correlate of irreversible disability in patients with multiple sclerosis. The availability of neurotrophins could influence the probability or rate of disease progression and the time of onset. The BDNF-Val66Met-polymorphism leads to altered intracellular transport and secretion of BDNF, and is thus a logical candidate for a gene that influences susceptibility and, more specifically, the clinical course of multiple sclerosis. In order to test this hypothesis we genotyped the polymorphism in 951 UK multiple sclerosis trio families, but found no evidence for association before (p=0.63) or after stratification for clinical course (p=0.73). PMID- 16046001 TI - Effects of sex hormones on costimulatory molecule expression in multiple sclerosis. AB - Sex hormones play a central role as modulators of immune responses and autoimmune diseases. We hypothesized that suppression of MS disease during pregnancy may be mediated by sex steroid hormones via regulation of costimulatory molecules such as CD40L or CD80/CD86 (B7-1/B7-2). We tested two sex hormones that are implicated in immune suppression during pregnancy: estriol and progesterone. We also examined whether this regulation is gender-specific or disease-related. PBMC from untreated relapsing remitting multiple sclerosis (RR MS) patients and controls were examined for expression of T cell and monocyte costimulatory molecules following mitogen stimulation in the presence or absence of sex hormones. In the absence of hormones, we confirmed that mitogen stimulation induced significantly more CD40L on the surface of CD4(+)T cells in MS patients compared to controls, and we extend these findings by showing there were no gender differences in induction of CD40L. Although supra-physiologic doses of hormones mildly suppressed CD40L expression on activated T cells, in vitro exposure to typical pregnancy-related physiologic doses of estriol or progesterone showed very little or no suppression of CD40L. On monocytes, neither estriol nor progesterone significantly altered the expression of CD80/CD86. These results suggest that physiologic doses of estriol or progesterone cannot alter CD40L on T cells or CD80/CD86 on monocytes sufficiently to explain the improvement observed in MS during pregnancy. Thus, although amelioration of MS and other autoimmune diseases during pregnancy is thought to be due to increased sex hormones, the present results do not support a role for suppression of costimulation via estriol or progesterone. PMID- 16046002 TI - Reply to: "M. Delaroche, R. Thiebaut, R. Dauman, Behavioural audiometry: protocols for measuring hearing thresholds in babies aged 4-18 months" [Int. J. Pediatr. Otorhinolaryngol. 68 (2004) 1233-1243]. PMID- 16046003 TI - Visualization of infectious pancreatic necrosis virus (IPNV) particles labeled with fluorescent probes. AB - Infectious pancreatic necrosis virus (IPNV) particles were labeled with SYBR Green I or a monoclonal antibody and FITC-conjugated secondary antibody and examined in a fluorescence microscope. Labeled viral particles were visualized in a narrow range of pixels. Comparing IPNV particles with fluorescent phage T4 virions, the former, as expected, were seen smaller in size. The method allows the rapid and accurate counting of viral particles both on filters and bound to the cell surface. In addition, IPNV particles can be specifically enumerated in the presence of other virions and the ratio between physical particles and virus infectivity can be easily calculated as well. PMID- 16046004 TI - A monoclonal antibody-based ELISA allows discrimination between responses induced by bovine herpesvirus subtypes 1 (BoHV-1.1) and 2 (BoHV-1.2). AB - Bovine herpesvirus type 1 (BoHV-1) has distinct subtypes according to genomic characterization. Immune responses induced by BoHV-1 subtype 1 (BoHV-1.1) are not distinguishable from those induced by BoHV-1 subtype 2 (BoHV-1.2) through conventional serological methods. In the present report, an enzyme linked immunosorbent assay is described that allows discrimination between immune responses in cattle immunized with either subtype, based on a monoclonal antibody that recognizes specifically the amino-terminal region of glycoprotein C (gC) on BoHV-1.1 strains, thus not reacting with BoHV-1.2a. The test displayed a sensitivity of 92%, specificity of 90% and a good correlation with serum neutralization tests on samples from BoHV-1.1-immunized calves (kappa = 0.799). The test may be useful to provide new insights into the roles played by each of these two subtypes in the epidemiology of BoHV-1 infections. PMID- 16046005 TI - STOP knockout and NMDA NR1 hypomorphic mice exhibit deficits in sensorimotor gating. AB - Schizophrenia is a chronic and debilitating disease which is thought to arise from a neuro-developmental disorder. Both the stable tubule-only polypeptide (STOP) protein and the N-methyl-D-aspartate (NMDA) NR1 subunit are involved in neuronal development and physiology. It has therefore been postulated that transgenic mice lacking either the STOP or the NMDAR1 gene would show a 'schizophrenic-like' phenotype. Here, STOP knockout and NMDA NR1 hypomorphic mice were assessed in a behavioural measure that can be used to detect schizophrenic like phenotypes: a change in sensorimotor gating, measured through prepulse inhibition (PPI). STOP knockout mice were further assessed in another measure of 'schizophrenic-like behaviour': hyperlocomotion. The PPI deficit exhibited by both the STOP knockout and NMDA knockdown mice could not be reversed by acute treatment with the atyptical antipsychotic, clozapine (1 mg/kg, i.p.) but the hyperlocomotion shown by the STOP knockout mice was reversed with the same acute dose of clozapine. PMID- 16046006 TI - Social and structural housing conditions influence the development of a depressive-like phenotype in the learned helplessness paradigm in male mice. AB - Structural and social factors are known to play a crucial role in the pathogenesis of depression. Since animal models of depression are a major tool to gain insights into the mechanisms involved in the pathophysiology of this disease it is important not only to exploit but also to be aware of factors that may affect these models. As housing represents a fundamental external factor, which is controversially debated to affect the animals' emotionality, this study aimed to investigate the impact of different social and structural housing conditions on the development of a depressive-like syndrome in the learned helplessness paradigm. Group housing in an impoverished environment led to an increased vulnerability in the learned helplessness paradigm. Groups that were housed enriched, however, were less helpless. Furthermore impoverished conditions did not increase the vulnerability in single housed animals. Regarding emotionality in the animals, basal anxiety was reduced and the exploration was enhanced by group housing and enriched environment. These results suggest that housing conditions significantly influence the outcome of learned helplessness studies. PMID- 16046007 TI - Sexual dimorphism in the vasopressin system: lack of an altered behavioral phenotype in female V1a receptor knockout mice. AB - Previous findings with an AVP V1a receptor knockout mouse (V1aRKO) demonstrate a significant role for this receptor in anxiety-like behavior in males. Here we report the lack of anxiety-like effects of the null mutation in female mice. V1aRKO females performed normally on all tests for anxiety-like behavior. This sex difference may be due to the sexual dimorphism in the extra-hypothalamic vasopressin system, with males having significantly more vasopressin fibers in this system. PMID- 16046008 TI - The screening of chemicals for juvenoid-related endocrine activity using the water flea Daphnia magna. AB - U.S. Environmental Protection Agency is charged with developing a screening and testing paradigm for detecting endocrine toxicity of chemicals that are subject to regulation under the Food Quality Protection and the Safe Drinking Water Acts. In this study, we developed and evaluated a screening assay that could be employed to detect juvenoid-related endocrine-modulating activity in an invertebrate species. Juvenoid activity, anti-juvenoid activity, and juvenoid potentiator activity of chemicals was assessed using the water flea Daphnia magna. Male sex determination is under the regulatory control of juvenoid hormone, presumably methyl farnesoate, and this endpoint was used to detect juvenoid modulating activity of chemicals. Eighteen chemicals were evaluated for juvenoid agonist activity. Positive responses were detected with the juvenoid hormones methyl farnesoate and juvenile hormone III along with the insect growth regulating insecticides pyriproxyfen, fenoxycarb, and methoprene. Weak juvenoid activity also was detected with the cyclodiene insecticide dieldrin. Assays performed repetitively with compounds that gave either strong positive, weak positive, or negative response were 100% consistent indicating that the assay is not prone to false positive or negative responses. Five candidate chemicals were evaluated for anti-juvenoid activity and none registered positive. Four chemicals (all trans-retinoic acid, methoprene, kinoprene, bisphenol A) also were evaluated for their ability to potentiate the activity of methyl farnesoate. All registered positive. Results demonstrate that an in vivo assay with a crustacean species customarily employed in toxicity testing can be used to effectively screen chemicals for juvenoid-modulating activity. PMID- 16046009 TI - Clathrin-independent endocytosis: new insights into caveolae and non-caveolar lipid raft carriers. AB - A number of recent studies have provided new insights into the complexity of the endocytic pathways originating at the plasma membrane of mammalian cells. Many of the molecules involved in clathrin coated pit internalization are now well understood but other pathways are less well defined. Caveolae appear to represent a low capacity but highly regulated pathway in a restricted set of tissues in vivo. A third pathway, which is both clathrin- and caveolae-independent, may constitute a specialized high capacity endocytic pathway for lipids and fluid. The relationship of this pathway, if any, to macropinocytosis or to the endocytic pathways of lower eukaryotes remains an interesting open question. Our understanding of the regulatory mechanisms and molecular components involved in this pathway are at a relatively primitive stage. In this review, we will consider some of the characteristics of different endocytic pathways in high and lower eukaryotes and consider some of the common themes in endocytosis. One theme which becomes apparent from comparison of these pathways is that apparently different pathways can share common molecular machinery and that pathways considered to be distinct actually represent similar basic pathways to which additional levels of regulatory complexity have been added. PMID- 16046010 TI - Self-rated health and social role as predictors for 6-year total mortality among a non-disabled older Japanese population. AB - We examined whether social role and self-rated health in an older population were predictors for 6-year total mortality among a non-disabled community-dwelling older population in Saku City, Nagano Prefecture, Japan, surveyed in 1992 and 1998. A total of 8090 men and women aged 65-99 years who reported no disability in performing activities of daily living (ADL) at the time of the survey in 1992 and provided information on their survival status at follow-up 6 years later were analyzed in this study. One dependent variable was survival status in 1998 and independent variables were various factors potentially associated with total mortality, which were obtained from a questionnaire survey at the baseline. During the 6-year interval, having poor self-rated health and poor social roles were identified as significant predictors for total mortality among both men and women. This study revealed that social role and self-rated health are independent predictors for 6-year total mortality for non-disabled Japanese aged 65 years or older. PMID- 16046011 TI - Key factors associated with the under-prescription of statins in elderly coronary heart disease patients: Results from the ELIAGE and ELICOEUR surveys. AB - INTRODUCTION: The reasons why statins are under-utilized in elderly patients remain poorly understood. The aim of this study was to identify the reasons given by cardiologist for the non-prescription of statins in elderly CHD patients. METHODS: Two cross-sectional pharmaco-epidemiological surveys were carried out among French cardiologists. The sample consisted of 1148 coronary patients aged 35 to 69 years and 1489 patients aged > or =70 years. Patients' risk factors, medical history, treatments, lipid values and the physicians' various motives for the non-prescription of statins were recorded. RESULTS: Patients not treated with statins reached 37% in the age-group > or =70 years and 14% in the age-group 35 69 years. The main reason given for statin non-prescription was the lack of a medical indication (2.5% of the age-group 35-69 years and 14% of the age-group > or =70 years). Among patients > or =70 years, the lack of indication was more often cited in the following conditions: 1) in very old patients (36% of lack of indication in the age-group >85 years vs. 10% in 70-75 years), 2) when lipid values were not available (20% when data were not available vs. 9%) and 3) when the patient had no prior history of myocardial infarction (MI) (20% when no history of MI vs. 7%). These factors were not associated with lack of indication among patients <70 years. History of intolerance or side effect was given for 1.3% and 14% of patients for each of the groups (35-69 and > or =70) and poor overall patient adherence was cited in 1% and 2%, respectively. CONCLUSION: The primary reason for the under-prescription of statins in elderly coronary patients is the perceived lack of indication, which stresses the need of extensive guidelines for prescription in elderly patients. Several factors associated with this perception seem to be specific to the elderly. PMID- 16046012 TI - A hidden cause of angina pectoris: when ABG analysis is the only helpful diagnostic tool. PMID- 16046013 TI - Impact of time past exercise testing on prognostic variables in heart failure. AB - BACKGROUND: Cardiopulmonary exercise testing (CPET) is considered a standard of care in heart failure (HF). The impact that duration of time post-evaluation has on the prognostic value of CPET, however, has not been explored. The purpose of this investigation was to examine the ability of peak oxygen consumption (VO(2)) and the minute ventilation-carbon dioxide production (VE/VCO(2)) slope to predict cardiac-related events at different time intervals post CPET. METHODS: Two hundred fifty-eight subjects diagnosed with HF underwent CPET. The ability of peak VO(2) and VE/VCO(2) slope to predict cardiac-related events without a time limit, one year post CPET and greater than one year post CPET was examined. RESULTS: Both peak VO(2) and VE/VCO(2) slope were significant predictors of cardiac-related mortality and hospitalization during the no time constraint and one year post CPET tracking scenarios (Hazard Ratio Range: 3.5-12.4, p<0.001). Peak VO(2) was no longer a significant predictor of mortality greater than one year post CPET. Generally, as time following CPET surpassed one year, the prognostic sensitivity of both variables slightly increased ( approximately 5%) while specificity sharply decreased ( approximately 20%). CONCLUSIONS: Both peak VO(2) and VE/VCO(2) slope are significant predictors of cardiac-related events in HF. However, the prognostic characteristics of these variables following CPET appear to be time-sensitive. PMID- 16046014 TI - A systems model of training for patients in phase 2 cardiac rehabilitation. AB - BACKGROUND: The aim of this study was to apply a systems model of training for athletes in patients with coronary artery disease (CAD) undergoing phase 2 cardiac rehabilitation. METHODS: Data from six patients with CAD undergoing 20 conventional training sessions were used to test this model. The method required daily training quantification using heart rate, and regular assessment of real exercise tolerance using a constant-duration test. Convolution of training quantity with real exercise tolerance was provided for every patient model exercise tolerance, by minimizing the residual sum of squares. RESULTS: The results showed that application of the systems model of training to the six patients resulted in a strong fit between real and model exercise tolerances: r(2) = 0.77, 0.79, 0.83, 0.84, 0.85 and 0.92, respectively (P < 0.05). DISCUSSION: Moreover, the systems model admitted for all patients corresponded to one first-order transfer function, which was fitness. This result was in contrast with the systems models reported in athletes and in a patient with CAD undergoing phase 3 cardiac rehabilitation which were found to include two first-order transfer functions: fitness and fatigue, simultaneously. CONCLUSION: The systems model of training of patients with CAD undergoing phase 2 suggested two practical consequences: first, the improvement of exercise tolerance is directly related to training, and consequently second, patients who enter immediately phase 3 training may prevent a decrease in their exercise tolerance. PMID- 16046015 TI - Effects of a high dose intravenous bolus amiodarone in patients with atrial fibrillation and a rapid ventricular rate. AB - BACKGROUND: Amiodarone, given as intravenous bolus has not yet been studied in patients with atrial fibrillation and a high ventricular rate. METHODS: One hundred consecutive patients with atrial fibrillation and a ventricular rate above 135 bpm were randomized to receive either 450 mg amiodarone or 0.6 mg digoxin given as a single bolus through a peripheral venous access. If the ventricular rate exceeded 100 bpm after 30 min, another 300 mg amiodarone or 0.4 mg digoxin were added. Primary endpoints of the study were the ventricular rate and the occurrence of sinus rhythm after 30 and 60 min. Secondary endpoints were blood pressure during the first hour after drug administration, and safety regarding drug induced hypotension, and phlebitis at the infusion site. RESULTS: Baseline heart rate was 144+/-19 in the amiodarone group and 145+/-15 in the digoxin group (p=0.72). Following amiodarone, heart rate was 104+/-25 after 30 min compared to 116+/-23 in the digoxin group (p=0.02) and 94+/-22 versus 105+/ 22 after 60 min (p=0.03). After 30 min, sinus rhythm was documented in 14 (28%) patients following amiodarone compared to 3 (6%) patients in the digoxin group (p=0.003), and after 60 min in 21 (42%) versus 9 (18%) patients (p=0.012). Asymptomatic hypotension was observed in 4 amiodarone treated patients, and superficial phlebitis in 1 patient. CONCLUSIONS: Amiodarone, given as an intravenous bolus is relatively safe and more effective than digoxin for heart rate control and conversion to sinus rhythm in patients with atrial fibrillation and a rapid ventricular rate. PMID- 16046016 TI - Impact of exercise training on muscle function and ergoreflex in Fontan patients: a pilot study. AB - BACKGROUND: Several studies have demonstrated persistent reduced exercise capacity in Fontan patients even after surgical intervention. The purpose of this study was to evaluate if the skeletal muscle function of these patients is abnormal, if it correlates with exercise tolerance and if it can be improved by exercise training. METHODS: We evaluated the functional capacity of seven patients who underwent Fontan procedure (age:16+/-5 years, mean+/-SD) and seven healthy children (19+/-7 years) paired for age, sex, height and weight. Evaluation included pulmonary evaluation, neuromuscular function and exercise tolerance. Secondly, an 8-week exercise training program was performed by five of these patients. RESULTS: The ergoreflex contribution to absolute diastolic blood pressure was higher (12.5+/-4.8 vs. 5.6+/-4.2 mmHg; p=0.04) in Fontan patients vs. healthy subjects whereas a trend was encountered regarding the ergoreflex contribution to absolute systolic blood pressure (9.0+/-7.0 vs. 0.4+/-9.0 mmHg; p=0.09). Furthermore, time to fatigue of the non-dominant forearm muscles was shorter in Fontan patients vs. healthy subjects (431+/-290 vs. 847+/-347 s; p=0.03). Following exercise training, there was a significant reduction of the ergoreflex contribution to absolute values of systolic blood pressure (9.8+/-0.9 vs. 0.3+/-2.7 mmHg; p<0.05). There was an association between muscle strength and VO2 peak in Fontan patients (upper limb: r=0.895; p<0.01; lower limb: r=0.838; p<0.05, respectively). CONCLUSIONS: Skeletal muscle function in Fontan patients is abnormal which may have an impact in the reduced exercise tolerance encountered in these patients. Exercise training may have beneficial impacts on the skeletal muscle function in this population. PMID- 16046017 TI - Prognostic value of resting end-tidal carbon dioxide in patients with heart failure. AB - BACKGROUND: Cardiopulmonary exercise testing (CPET) variables provide valuable prognostic information in the heart failure (HF) population. The purpose of the present study is to assess the ability of resting end-tidal carbon dioxide partial pressure (PETCO2) to predict cardiac-related events in patients with HF. METHODS: 121 subjects diagnosed with compensated HF underwent CPET on an outpatient basis. Mean age and ejection fraction were 49.3 years (+/-14.7) and 28.4% (+/-13.4), respectively. Resting P(ET)CO2 was determined immediately prior to the exercise test in the seated position. Peak oxygen consumption (VO2) and the minute ventilation-carbon dioxide production (VE/VCO2) slope were also acquired during CPET. RESULTS: There were 41 cardiac-related hospitalizations and 9 cardiac-related deaths in the year following CPET. Mean resting P(ET)CO2, peak VO2 and VE/VCO2 slope were 34.1 mmHg (+/-4.6), 14.5 ml*kg(-1)*min(-1) (+/-5.1) and 35.9 (+/-8.7) respectively. Univariate Cox regression analysis revealed that resting P(ET)CO2 (Chi-square=28.4, p<0.001), peak VO2 (Chi-square=21.6, p<0.001) and VE/VCO2 slope (Chi-square=54.9, p<0.001) were all significant predictors of cardiac related events. Multivariate Cox regression analysis revealed resting P(ET)CO2 added to the prognostic value of VE/VCO2 slope in predicting cardiac related events (residual Chi-square=4.4, p=0.04). Peak VO2 did not add additional value and was removed (residual Chi-square=3.2, p=0.08). CONCLUSIONS: These results indicate a resting ventilatory expired gas variable possesses prognostic value independently and in combination with an established prognostic marker from the CPET. Resting P(ET)CO2 may therefore be a valuable objective measure to obtain during both non-exercise and exercise evaluations in patients with HF. PMID- 16046018 TI - A comparative study of markers of inflammation for the assessment of cardiovascular risk in patients presenting to the emergency department with acute chest pain suggestive of acute coronary syndrome. AB - BACKGROUND: The role of inflammation in the pathogenesis of acute coronary syndrome (ACS) is established. Little is known however, regarding the use of inflammatory markers as predictors of future cardiovascular events in patients presenting to the emergency department (ED) with suspected ACS. HYPOTHESIS: To assess whether biomarkers that predict cardiovascular risk in apparently healthy individuals and coronary artery disease patients are useful predictors of future cardiovascular events in patients presenting to the ED with chest pain suggestive of ACS. METHODS: We compared the abilities of serum C-reactive protein (hs-CRP), albumin and leukocyte count to identify subjects with ACS and those who are at high risk of developing events during a 30-day follow-up. RESULTS: 144 patients (mean age 62+/-13 years, 45 female) presenting to the ED <3 h after the onset of symptoms suggestive of ACS were evaluated. Final hospital diagnoses were non ischemic chest pain in 43 (30%) and ACS in 101 (70%) patients. Patients with ACS had significantly higher leukocyte count (p<0.0001) and hs-CRP levels (p<0.02) and lower albumin concentrations, compared to patients with NICP (p<0.0001). Lower albumin concentrations (p=0.03) and hs-CRP (p=0.049) were predictors of recurrent events at 30 days. On multivariate analysis, however, only leukocyte count was a predictor of ACS (OR 20.9; 95% CI: 3.7-19.5; p=0.01) and high hs-CRP levels were a predictor of clinical outcome (OR 2.8; 95% CI: 1.5-5.2; p=0.001). CONCLUSIONS: Leukocyte count is an independent predictor of ACS in patients presenting to the ED with chest pain suggestive of ACS and high hs-CRP levels are an independent predictor of clinical outcome in ACS patients. PMID- 16046019 TI - The value of genetic information in selecting dairy replacements. AB - The objective of this study was to empirically determine the economic value of genetic information in the selection of dairy replacements, and assess whether this value was sufficient to prompt producers to select replacements on this basis. The data set consisted of 1982 Michigan Holstein replacements in 115 herds. Each herd had a minimum of 10 replacements that were born in the last 6 months of 1992 and calved within the last 6 months of 1994. The data for each replacement included the estimated breeding value (EBV) for milk at the beginning and end of the rearing period, and the estimated lifetime profit corrected for the opportunity cost of postponed replacement (ELPCOC). The replacement selection decision for a profit-maximizing dairy producer selecting 70 or 80% of the replacements was modeled. We modeled three methods of selection: genetic, random and ex poste. Genetic selection was evaluated using the EBV milk available at the beginning or end of the rearing period. For each herd, the profit associated with each of the three methods of selection was simulated. The value of the genetic information and perfect information were the differences in herd profits of genetic selection and ex poste selection relative to random selection, respectively. The difference in value of the genetic information between the end of the rearing period and the beginning of the rearing period was the increase in value of the genetic information due to updating. The value of information was calculated as the average herd profit per replacement. The value of the genetic information ranged from 22 dollars/replacement to 30 dollars/replacement and was statistically greater than zero at a 95% confidence level. It is unclear whether this value is sufficient to prompt producers to select replacements on the basis of EBV milk as has been recommended. The negative value of EBV milk (from the end of the rearing period when selecting 80% of the replacements) for 32 herds was consistent with the noisiness of the genetic estimates as messages of ELPCOC. The increased value of the genetic information due to updating was approximately 5 dollars/replacement. This increased value is likely insufficient to warrant delaying replacement selection decisions solely to obtain the updated information. The value of EBV milk was approximately 4 dollars/replacement higher when selecting 70% of the replacements versus 80%. The genetic information captured between 15% (selecting 70% at the beginning of the rearing period) and 20% (selecting 80% at the end of the rearing period) of the value of perfect information. PMID- 16046020 TI - Induction of CTGF by TGF-beta1 in normal and radiation enteritis human smooth muscle cells: Smad/Rho balance and therapeutic perspectives. AB - BACKGROUND AND PURPOSE: Transforming Growth Factor beta1 (TGF-beta1) and its downstream effector Connective Tissue Growth Factor (CTGF/CCN2), are well known fibrogenic activators and we previously showed that the Rho/ROCK pathway controls CTGF expression in intestinal smooth muscle cells isolated from patients with delayed radiation enteritis. The aim of the present work was to investigate the balance between Smad and Rho signalling pathways in the TGF-beta1 CTGF induction and modulation of radiation-induced fibrogenic differentiation after addition of pravastatin, an inhibitor of Rho isoprenylation. PATIENTS AND METHODS: Primary human smooth muscle cells isolated from normal (N-SMC) or radiation enteritis (RE SMC) biopsies were incubated with TGF-beta1 (10 ng/ml). Induction of CTGF, as well as nucleo-cytoplasmic distribution of phospho-Smad2/3, Smad2/3 and Smad4 were analysed by Western blot and immunocytochemistry. Smad DNA binding was assessed by EMSA and Rho activation was measured by pull-down assay. RESULTS: After TGF-beta1 addition, Smads were translocated to the nucleus in both cell types. Nuclear accumulation of Smad as well as their DNA-binding activity were higher in N-SMC than in RE-SMC, whereas the opposite was observed for Rho activation, suggesting a main involvement of Rho pathway in sustained fibrogenic differentiation. This hypothesis was further supported by the antifibrotic effect observed in vitro after cell treatment with pravastatin (i.e. decreased expression of CTGF, TGF-beta1 and Collagen Ialpha2). CONCLUSIONS: Our results suggest that TGF-beta1-induced CTGF transactivation mainly depends on the Smad pathway in N-SMC, whereas in RE-SMC, Smad and Rho pathways are involved. Inhibition of Rho activity by pravastatin alters fibrogenic differentiation in vitro which opens up new therapeutic perspectives. PMID- 16046021 TI - Incidence of trichothecenes and zearalenone in poultry feed mixtures from Slovakia. AB - A total of 50 samples of poultry feed mixtures of Slovakian origin were analyzed for eight toxicologically significant Fusarium mycotoxins, namely zearalenone (ZON), A-trichothecenes: diacetoxyscirpenol (DAS), T-2 toxin (T-2) and HT-2 toxin (HT-2) and B-trichothecenes: deoxynivalenol (DON), 3-acetyl-deoxynivalenol (3 ADON), 15-acetyl-deoxynivalenol (15-ADON) and nivalenol (NIV). The A trichothecenes and the B-trichothecenes were detected by means of high pressure liquid chromatography with tandem mass spectrometry detection (HPLC-MS/MS) and gas chromatography electron capture detection (GC-ECD), respectively. Reversed phase-high performance liquid chromatography with a fluorescence detector (RP HPLC-FLD) was used for ZON detection. The most frequent mycotoxin detected was T 2, which was found in 45 samples (90%) in relatively low concentrations ranging from 1 to 130 microg kg(-1) (average 13 microg kg(-1)), followed by ZON that was found in 44 samples (88%) in concentrations ranging from 3 to 86 microg kg(-1) (average 21 microg kg(-1)). HT-2 and DON were detected in 38 (76%) and 28 (56%) samples, respectively, in concentrations of 2 to 173 (average 18 microg kg(-1)) for HT-2 and 64 to 1230 microg kg(-1) sample (average 303 microg kg(-1)) for DON. The acetyl-derivatives of DON were in just four samples, while NIV was not detected in any of the samples investigated. In as many as 22 samples (44%), a combination of four simultaneously co-occurring mycotoxins, i.e. T-2, HT-2, ZON and DON, was revealed. Despite the limited number of samples investigated during this study poultry feed mixtures may represent a risk from a toxicological point of view and should be regarded as a potential source of the Fusarium mycotoxins in Central Europe. This is the first reported study dealing with zearalenone and trichothecene contamination of poultry mixed feeds from Slovakia. PMID- 16046022 TI - Development and characterization of a monoclonal antibody directed against human telomerase reverse transcriptase (hTERT). AB - Telomerase activity plays an important role in the two complementary processes of cellular immortalization and senescence. This enzyme is active in almost all tumors, but also in inflammatory and many normal proliferating cells. Therefore, the main limits of molecular determinations, such as telomeric repeat amplification protocol assay is that they are not able to discriminate between the enzymatic activity of tumor and normal cells. The most appropriate technique for this would be immunohistochemical determination using monoclonal antibodies. Very few monoclonal antibodies (Mabs) directed against the human telomerase reverse transcriptase (hTERT) are commercially available and in the present study, we developed a new Mab directed against this protein (TERT-3 36-10) to investigate the possibility of detecting immunoreactivity to this Mab by immunohistochemical and flow cytometric approaches. Immunohistochemical determination showed a lack of reactivity to the Mab in highly differentiated striated muscle tissue, a variable reactivity in dysplastic cervical epithelial tissue and similar and widespread immunoreactivity in cell lines and clinical tumors. Furthermore, we demonstrated the ability of this Mab to inhibit enzyme activity in cell extract from MCR bladder tumor cell line. PMID- 16046023 TI - The role of cavitation in acoustically activated drug delivery. AB - Pluronic P105 micelles are potential candidates as chemotherapy drug delivery vehicles using ultrasonic stimulation as a release trigger. Acoustic power has been previously shown to release two anthracycline agents from these polymeric carriers. In this study, an ultrasonic exposure chamber with fluorescence detection was used to examine the mechanism of doxorubicin release from P105 micelles. Acoustic spectra were collected and analyzed, at the same spatial position as fluorescence data, to probe the role of cavitation in drug release. Our study showed a strong correlation between percent drug release and subharmonic acoustic emissions, and we attribute the drug release to collapse cavitation that perturbs the structure of the micelle and releases drug. PMID- 16046024 TI - Depressive symptoms of type 2 diabetics treated with insulin compared to diabetics taking oral anti-diabetic drugs: a Korean study. AB - OBJECTIVE: The purpose of this study was to examine the depressive symptoms of type 2 diabetic patients who were treated with insulin compared to those diabetics treated with oral anti-diabetic drugs in Korea. METHODS: A total 204 outpatients with type 2 diabetes were invited to complete a questionnaire using the Beck depression inventory (BDI) to measure depressive symptoms. Age, gender, body mass index, serum lipid profile, and a social history including marital status, occupation and educational background were also assessed. The presence of diabetic complications was evaluated by examining the patients' medical records. Diabetic patients who were not treated with anti-diabetic drugs were excluded. All the study subjects were classified into two groups based on their mode of therapy: the oral drug group and insulin group. The insulin group included patients treated with insulin-oral drug combinations as well as those treated solely with insulin. RESULTS: Overall, 32.4% of our diabetic subjects showed depressive symptoms with the criterion being a BDI score > or = 16. Compared to the oral drug group, the insulin group showed a significantly higher frequency of depressive symptoms (insulin group, 48.0%; oral drug group, 27.3%; p<0.01) and higher BDI scores (insulin group, 16.6+/-10.7; oral drug group, 12.6+/-8.7; p<0.01). Moreover, after an adjustment for social factors, the degree of hyperglycemia and the presence of diabetic complications, the insulin group showed a significantly higher frequency of depression (odds ratio 4.38, 95% CI 1.66-11.6, p=0.003), compared to the oral drug group. CONCLUSIONS: The data showed that insulin treatment is related to the presence of depressive symptoms, and the importance of more careful psychological support of Korean insulin treated type 2 diabetic patients is strongly suggested. PMID- 16046025 TI - Divining the role of liver biopsy in hepatitis C. PMID- 16046026 TI - The impact of decentralised drug-budgets in Sweden - a survey of physicians' attitudes towards costs and cost-effectiveness. AB - In Sweden decentralised drug-budgets at health-care facility levels were introduced in 1997 in an attempt to contain increasing pharmaceutical expenditures. This paper reports the findings of a postal survey which investigates whether decentralised drug-budgets according to a so-called primary care based model in Swedish health care have led to increased cost awareness and changed attitudes towards cost-minimisation and cost-effectiveness as decision making criteria among physicians. In particular, it was investigated whether there were differences in this respect between general practitioners (GPs) and specialists. The postal survey was sent to 1,520 Swedish physicians from a stratified sample of Swedish county councils. A total of 738 physicians responded (response rate 49%). Statistical analysis was performed of logistic regression analysis and independent or paired samples t-tests. The results suggest that GPs have a higher degree of cost awareness than specialists. Physicians with experience of decentralised drug-budgets have a higher degree of cost awareness than other physicians. However, the rating of the top four decision-making criteria; therapeutic effects, side effects, compliance and cost-effectiveness, were not significantly different when comparing GPs against specialists, and physicians practising in county councils with decentralised drug-budgets against other physicians. The main barriers to considering costs to a greater extent were perceived difficulties in switching drugs and a fear among physicians of losing credibility among patients. In conclusion, decentralisation of drug-budgets according to the Swedish primary-care based model increases cost awareness, especially among GPs. Such responsibility, however, does not create strong incentives for physicians to reconsider the importance of cost-effectiveness in relation to other decision-making criteria when prescribing. Parallel interventions are needed to meet the objective of cost-effective prescribing. PMID- 16046027 TI - Social capital and farmer's willingness-to-join a newly established community based health insurance in rural China. AB - In 2002, China announced a new funding strategy that would reestablish community based health insurance (CHI) in rural areas, whereby the Chinese government will entice farmers' participation by providing each participant an annual subsidy of 10-20 Yuan (US 1.25-2.50 dollars). However, there is no evidence demonstrating how many farmers would be willing-to-join (WTJ) such newly developed government subsidized voluntary-based CHI scheme and what factors influence farmers' willingness-to-join. In this study, we examine the probability of farmers' willingness-to-join such CHI under the different scenarios of government subsidy and individual contribution, and also explore factors that influence farmers' willingness-to-join with the emphasis on social capital. The study is based on data collected from a 2002 household survey conducted in Fengsan Township, located in China's Guizhou Province. Logistic regression is used in the analysis. The findings from this study show that even with the government subsidy to the premium, the probability of WTJ the new voluntary-based CHI only reach 50%. The results also indicate that community level social capital, as measured by reciprocity index, and individual level social capital, as measured by trust index, are significantly and positively associated with the probability of farmers' WTJ newly developed government subsidized CHI. Policy recommendations have been made based on those findings. PMID- 16046028 TI - Both the phosphoinositide and receptor binding activities of Dab1 are required for Reelin-stimulated Dab1 tyrosine phosphorylation. AB - Reelin-stimulated tyrosine phosphorylation of the Dab1 adaptor protein is required during brain development for Reelin-dependent neuron positioning in the cerebral cortex and various other laminated regions. Dab1 contains an amino terminal PTB/PI domain through which it can bind to Reelin receptors and membrane phosphoinositides. The relative contributions of these binding activities were unknown. Here, we identify a mutation in the PTB domain of Dab1 that inhibits membrane localization without inhibiting receptor binding. In neurons, this mutation reduces both basal and Reelin-stimulated Dab1 tyrosine phosphorylation. In contrast, a mutation that inhibits receptor binding reduces Reelin-stimulated but not basal tyrosine phosphorylation. These results support a model in which phospholipids recruit Dab1 to membranes but do not play a direct role in relaying the Reelin signal, while direct Dab1-receptor interaction is responsible for relaying the Reelin signal but not for membrane recruitment. PMID- 16046029 TI - Effectiveness of transbronchial needle aspiration in the diagnosis of exophytic endobronchial lesions and submucosal/peribronchial diseases of the lung. AB - The role of transbronchial needle aspiration (TBNA) in diagnosing endobronchial lung cancers has not been elucidated. The definitive combination of procedures that offers the best diagnostic yield following fiberoptic bronchoscopy remains controversial. This study was designed to investigate the diagnostic yield of transbronchial needle aspiration and other cytologic and histologic diagnostic procedures (i.e., forceps biopsy, brushing, and washing) and to assess the optimal combination for diagnosing endobronchial lung cancers. This prospective study included 95 patients presenting with visible tumors detected during bronchoscopic procedure as either an exophytic endobronchial lesion (EEL) or submucosal-peribronchial disease (SPD). Transbronchial needle aspiration, forceps biopsy, brushing, and washing were performed in all patients, and 91 patients were diagnosed. Rates of positive results were 75.8% for needle aspiration, 71.6% for forceps biopsy, 61.1% for brushing, and 32.6% for washing. Needle aspiration was used as the sole diagnostic method in 11, forceps biopsy was the sole diagnostic method in 5, and brushing was the sole diagnostic method in 4 patients. Washing was not used as the sole diagnostic method in any case. Forceps biopsy yielded the highest diagnostic rate for an EEL (86.4%); however, when compared with needle aspiration (77.9%), no significant difference was observed between these two procedures (P = 0.302). In patients with a diagnosis of SPD, needle aspiration was determined to be the sole diagnostic method in eight patients. In this group of patients, the highest rate of diagnosis was achieved with needle aspiration (72.2%), and when compared with forceps biopsy (47.2%), a significant difference between the two procedures (forceps biopsy versus needle aspiration) was observed (P = 0.049). By adding transbronchial needle aspiration to the conventional diagnostic methods (forceps biopsy, brushing, and washing), the rate of diagnosis increased from 82.1% to 95.8% (P = 0.001), and in patients with a diagnosis of SPD, this rate increased from 69.4% to 94.4% (P = 0.008). In patients with a diagnosis of an EEL, addition of needle aspiration led to an increase in diagnostic yield but this difference was not statistically significant (89.8% versus 96.6%, P = 0.250). In endobronchial lung cancers, transbronchial needle aspiration is a safe method that can be used together with conventional diagnostic procedures to increase the diagnostic yield and should be considered a valuable diagnostic tool, particularly in cases of SPD. The highest rate of diagnostic yield in this study was obtained using a combination of forceps biopsy, transbronchial needle aspiration, and brushing; washing did not contribute to this high rate. PMID- 16046030 TI - Normal neuroanatomical variation due to age: the major lobes and a parcellation of the temporal region. AB - We used high-resolution MRI to investigate gray and white matter aging in the major lobes of the cerebrum (frontal, parietal, temporal, occipital) and the major sectors of the temporal lobe (temporal pole, superior temporal gyrus, infero-temporal region, parahippocampal gyrus, amygdala, hippocampus). Subjects included 87 adults between the ages of 22 and 88 years. Regions of interest were hand-traced on contiguous 1.5mm coronal slices. For the cerebrum in general, gray matter decreased linearly with age, resulting in a decline of about 9.1-9.8% between the ages of 30 and 70 years, and a decline of 11.3-12.3% by the age of 80. In contrast, white matter volume increased until the mid-50s, after which it declined at an accelerated rate. At 70 years, white matter volume was only 5.6 6.4% less than at 30 years, but by age 80, a cubic regression model predicted that the decrease would be 21.6-25.0%. Multivariate analyses indicate that the frontal gray matter was most strongly associated with age, while occipital gray and white matter were least associated. Reduction in volume in the hippocampus was best modeled by a cubic regression model rather than a linear model. No sex differences in aging were found for any regions of interest. PMID- 16046031 TI - Telomere shortening in T lymphocytes of older individuals with Down syndrome and dementia. AB - Telomere shortening has been recently correlated with Alzheimer's disease status. Therefore, we hypothesized that a possible association might exist for adults with Down syndrome (DS). Using blind, quantitative telomere protein nucleic acid FISH analyses of metaphase and interphase preparations from 18 age-matched trisomy 21 female study participants with and without dementia, we have observed increased telomere shortening in adults with DS and dementia (p < .01). From this initial study, we conclude that telomere shortening is associated with dementia in this high-risk population and suggest that additional research may show that telomere shortening may be a biological marker of dementia status. PMID- 16046032 TI - Assessment of PINK1 (PARK6) polymorphisms in Finnish PD. AB - Recent data has demonstrated that mutations in PINK1, encoding PTEN-induced kinase 1, are a cause of early onset recessive parkinsonism (PARK6 locus). Common variability in genes implicated in hereditary forms of parkinsonism may be a predisposing factor in sporadic Parkinson's disease (PD). We analyzed whether six different genetic variants within and surrounding PINK1 contribute to the risk of sporadic PD in a Finnish case-control series. Our results indicate that this gene does not play a major role in the genetic predisposition to PD in this population. PMID- 16046033 TI - [Cerebral venous thrombosis]. AB - INTRODUCTION: Although more rare than arterial thrombosis, cerebral venous thrombosis are a non-negligible cause of stroke. Characterised by the large diversity of clinical presentations and etiologies, they have a much better prognosis than arterial stroke. The evolution remains unforeseeable, with a non negligible proportion of worsening at the acute phase and diagnosis must be early to begin as soon as possible the treatment, which is at present based on heparin therapeutics. CURRENT KNOWLEDGE AND KEY POINTS: Neuroimaging examinations are essential for diagnosis of CVT. MR Imaging with MR venography is the key procedure. New sequences are on evaluation in CVT bringing some physiopathogical arguments (Diffusion weighted imaging) or help for diagnosis (with T2* MRI sequence). If D-dimers dosage is helpful for diagnosis of deep venous thrombosis, its interest remains to be determined during CVT. CONCLUSION: CVT diagnosis is a challenge for the clinician. Because of the multiple causes and favorising factors, CVT are at the convergence of many specialties and could thus benefit of each one contribution for better understanding the physiopathology, improving earlier diagnosis or identifying the severe forms that could require right away more aggressive treatments than heparin. The interest of local thrombolysis or thrombectomy remains to be determined in an international randomised study. PMID- 16046034 TI - Nutritional assessment and management in hospitalised patients: implication for DRG-based reimbursement and health care quality. AB - INTRODUCTION: Malnutrition is associated with a higher morbidity resulting in an increased need for medical resources and economic expenses. In order to ensure sufficient nutritional care it is mandatory to identify the effect of malnutrition and nutritional care on direct cost and reimbursement. The primary aim of this study was to evaluate the economic effect of a nutritional screening procedure on the identification and coding of malnutrition in the G-DRG system. METHODS: All G-DRG relevant parameters of 541 consecutive patients at a gastroenterology ward were documented. Moreover, all patients were screened for malnutrition by a dietician according to the subjective global assessment (SGA). Patients were then grouped into the appropriate G-DRG and the effective cost weight (CW) was calculated. RESULTS: Ninety-two of 541 patients (19%) were classified malnourished (SGA B or C). Recognition of malnutrition increase from 4% to 19%. Malnourished patients exhibited a significantly increased length of hospital stay (7.7+/-7 to 11+/-9, P<0.0001). In 26/98 (27%) patients, the coding of malnutrition was considered relevant by grouping and resulted in a rise of DRG benefit. Mean case mix value and patients' complexity and comorbidity level (PCCL) increased after including malnutrition in the codification (CV 1.53+/-2.9 to 1.65+/-2.9, P=0.001 and PCCL 2.69+/-1.4 to 3.47+/-0.82, P<0.0001). The reimbursement increase by 360/malnourished patient or an additional reimbursement of 35280 (8.3% of the total reimbursement for all patients of 423186). Nutritional support in a subgroup of 50 randomly selected patients resulted in additional costs of 10268 . Forty-four of these patients (86%) were classified malnourished (32 SGA B and 12 SGA C). However, the subsequent reimbursement covered only approximately 75% of the expenses (7869), but did not include the potential financial benefits resulting from clinical interventions. CONCLUSION: Malnourished patients can be detected with a structured assessment and documentation of nutritional status and this is partly reflected in the G-DRG/ICD 10 system. In addition to increasing direct health care reimbursement, nutritional screening and intervention has the potential to improve health care quality. PMID- 16046035 TI - Influenza vaccination coverage rates in five European countries-a population based cross-sectional analysis of two consecutive influenza seasons. AB - INTRODUCTION: Influenza continues to be a considerable health problem in Europe. Vaccination is the only preventive measure, reducing mortality and morbidity of influenza in all age groups. OBJECTIVES: The objective of this survey was to assess the level of influenza vaccination coverage during two consecutive influenza seasons (2002/2003 and 2003/2004) in six European countries, to understand the driving forces and barriers to vaccination and to determine vaccination intentions for the following winter. METHODS: We conducted a random sampling, telephone-based household survey among non-institutionalised individuals representative of the population aged 14 and over. The surveys used the same questionnaire for two consecutive winters: 2002/2003 and 2003/2004 data were used for Germany, Italy, Spain and the United Kingdom. 2001/2002 and 2002/2003 data were used for France. The data were subsequently pooled. Four target groups were determined for analysis: (1) persons aged 65 and over; (2) people working in the medical field; (3) persons suffering from chronic illness and (4) a group composed of persons aged 65 and over or working in the medical field or suffering from a chronic illness. RESULTS: The overall sample consisted of 20,118 individuals. The influenza vaccination coverage rate increased from 21.3% in the first season to 23.2% in the second season. The increase in coverage is statistically significant (p=0.01). The most frequent reasons for being vaccinated given by vaccines were: influenza, considered to be a serious illness which people wanted to avoid (55.8%), having received advice from the family doctor or nurse to be vaccinated (55.2%) and not wanting to infect family and friends (36.1%). Reasons for not being vaccinated mentioned by people who have never been vaccinated were: not expecting to catch influenza (40.4%), not having considered vaccination before (33.3%) and not having received a recommendation from the family doctor to be vaccinated (27.3%). Options encouraging influenza vaccination are: recommendation by the family doctor or nurse (53.1%), more available information on the vaccine regarding efficacy and tolerance (32.1%) and more information available about the disease (26.7%). Adjusted odds ratios for target group vaccination were between 3.6 (Germany) and 13.7 (UK). Vaccination rates among healthcare workers were generally very low. Adjusted odds ratios were between 0.7 (Germany) and 1.5 (Spain). CONCLUSION: The vaccination coverage during the second season increased in comparison to the first season. The family doctor is the most important source of encouragement for people to be vaccinated against influenza. It seems that the public would be more likely to be vaccinated if they had more information on the efficacy and tolerance of the vaccine, as well as the disease. We, therefore, suggest that family doctors be better informed on influenza vaccine and the disease itself, so that they can actively inform their patients on these topics. PMID- 16046036 TI - Cost-effectiveness of varicella vaccination of healthcare workers. AB - OBJECTIVE: Although transmission of VZV is recognized as an important cause for morbidity in health care workers (HCWs), there is no general agreement on the recommended immunization policy. This study aimed to evaluate several of such prevention policies in economic terms. SETTINGS: Analysis of the cost per avoided future varicella cases among HCWs in Israel. METHODS: A cost-effectiveness analysis was performed by comparing the cost per avoided case of varicella among a theoretical cohort of 63,353 physician and nurses aged less than 45 years in Israel. Four policies were examined: (a) do nothing (status quo); (b) vaccination of susceptible workers using VZV serotesting; with (c) or without anamnestic history of chickenpox; and (d) presumptive mass vaccination of all eligible workers. A Markov-based model was developed using data from our recent seroepidemiological study in target population and from the literature. RESULTS: Screening and vaccination of susceptible workers using anamnestic selection is expected to reduce future cases, within 20 years since vaccination, from 58.3 to 33.0 with an incremental cost of 23,713 US dollars per avoided cases. Using only serological tests to detect susceptible workers would prevent additional 5.7 cases with an incremental cost of 206,692 US dollars per avoided case. Vaccinating all HCWs without serotesting, raises the costs markedly, with almost identical effectiveness, resulting in an incremental cost of 10.4 million US dollars per avoided case. Sensitivity analyses do not alter the ranking of the options. CONCLUSION: From the employer's perspective, routine varicella vaccination program for HCWs with or without selection of susceptible workers is extremely expensive compared to other high-cost practiced approaches. Substantial reduction in cost of vaccination may alter this conclusion. PMID- 16046037 TI - Candidacy of LPS-based glycoconjugates to prevent invasive meningococcal disease: developmental chemistry and investigation of immunological responses following immunization of mice and rabbits. AB - Glycoconjugates were prepared by covalently linking the immunogenic protein carrier CRM(197) to O-deacylated lipopolysaccharide (LPS) derived from Neisseria meningitidis (strain H44/76), immunotype L3 galE LPS. This mutant strain elaborates a truncated LPS structure that displays immunological epitopes characteristic of 76% of Group B meningococcal (NmB) strains. CRM(197) was covalently linked either to the reducing glucosamine residue of the lipid A region of the O-deacylated LPS or to a 2-keto-3-deoxy-octulosonic acid (Kdo) residue in the inner core region of the O-deacylated LPS. In both rabbits and mice a much stronger IgG response to the immunising antigen was generated in those animals that received conjugates linked via the lipid A region. Sera from mice that were immunized with these conjugates were assayed for their reactivity with LPS, both mutant and wild-type, of several homologous and heterologous NmB strains. Sera obtained from mice immunized with conjugates in which the carrier protein was linked via the Kdo moiety were only able to react with O-deacylated, but not fully acylated (native), LPS from the homologous strain. However, sera obtained from mice that were immunized with conjugates, in which the carrier protein was coupled to the lipid A region, reacted predominately with inner core epitopes that contained phosphoethanolamine at the same 3-position of the distal heptose residue (HepII) of the inner core LPS as was present on the immunising antigen. Additionally it was observed that sera from rabbits immunised with lipid A linked conjugates, unlike the mice responses, were generally not as specific for LPS antigens that contained phosphoethanolamine at the same 3-position as was present on the immunising antigen, but showed a broader inner core recognition, whereas those rabbits that received the Kdo-linked conjugates gave only a very weak non-specific response to all immunotypes. Finally, the sera from two out of six mice that had received lipid A linked conjugates had bactericidal activity against L3 wild-type NmB strain 8047 and one of these was able to passively protect against meningococcal infection in an infant rat model. This study demonstrates evidence towards the proof-in-principle that by using Nm inner core LPS conjugates coupled via the lipid A region with an intact phosphoethanolamine at the O-3 position of the HepII of the inner core LPS, it is possible to elicit functional and protective antibodies against meningococcal infection. PMID- 16046038 TI - Conflicts in how interpreters manage their roles in provider-patient interactions. AB - Interpreters face challenges because of the various role expectations that others have placed on them and then adopt specific strategies to manage these conflicts. This study examines the conflicts in medical interpreters' role performances, the sources of these conflicts, and interpreters' strategies for resolving conflicts. It is based on in-depth interviews with 26 medical interpreters from 17 languages in the Midwestern area of the USA. The results showed that interpreters experienced four sources of conflicts in their role performances: (a) others' communicative practices, (b) changes in participant dynamics, (c) institutional constraints, and (d) unrealistic role expectations. To resolve conflicts, interpreters justified their roles by identifying the source and location of an assignment, (re)defining the relationships and identities of the provider and the patient, and adopting specific communicative strategies. This study highlights the importance of speaker and contextual factors on interpreters' communicative strategies and management of role conflicts. PMID- 16046039 TI - Representations of mad cow disease. AB - This paper examines the reporting of the story of Bovine Spongiform Encephalopathy (BSE) and its human derivative variant Creutzfeld-Jacob Disease (vCJD) in the British newspapers. Three 'snapshots' of newspaper coverage are sampled and analysed between the period 1986 and 1996 focusing on how representations of the disease evolved over the 10-year period. Social representations theory is used to elucidate how this new disease threat was conceptualised in the newspaper reporting and how it was explained to the UK public. This paper examines who or what was said to be at risk from the new disease, and whether some individuals or groups held to blame for the diseases' putative origins, the appearance of vCJD in human beings, and its spread. PMID- 16046040 TI - Heroic frames: discursive constructions around the requested death movement in Australia in the late-1990s. AB - This paper focuses on a critical development in the life of the requested death movement [McInerney, F. (2000). "Requested Death": A new social movement. Social Science & Medicine, 50(1), 137-54.], that being the passage of the Northern Territory of Australia's Rights of the Terminally Ill (ROTI) Act 1995. This legislation, for the first time anywhere in the world, allowed for lawful euthanasia and physician-assisted suicide, thereby fulfilling key requirements of the movement. Taking a constructionist perspective, I analyzed discursive representations of dying, death and medicine in selected Australian print media during this time period (1995-1997). The media's predilection for reporting dramatic and unusual death coincided with the movement's construction of contemporary dying as horrific, intractable, and intolerable. Across all analyzed publications and genres, an heroic discourse was found to be a dominant influence, couched within a dramatic framing that served to reinforce many of the claims of the requested death movement. The framing of requested death activists as heroes, and of requested death itself as a redeeming and transforming act for those seeking it, were preeminent in press portrayals. The dominance of this heroic discourse suggests that such media and movement frames worked in tandem to both resonate with and reinforce popular Australian notions of terminal illness and dying in the late 20th century. PMID- 16046041 TI - Gender imbalance in infant mortality: a cross-national study of social structure and female infanticide. AB - Sex differentials in infant mortality vary widely across nations. Because newborn girls are biologically advantaged in surviving to their first birthday, sex differentials in infant mortality typically arise from genetic factors that result in higher male infant mortality rates. Nonetheless, there are cases where mortality differentials arise from social or behavioral factors reflecting deliberate discrimination by adults in favor of boys over girls, resulting in atypical male to female infant mortality ratios. This cross-national study of 93 developed and developing countries uses such macro-social theories as modernization theory, gender perspectives, human ecology, and sociobiology/evolutionary psychology to predict gender differentials in infant mortality. We find strong evidence for modernization theory, human ecology, and the evolutionary psychology of group process, but mixed evidence for gender perspectives. PMID- 16046042 TI - 'Between the demands of truth and government': health practitioners, trust and immunisation work. AB - Applied and theoretical work on health governance and governmentality has often not engaged with the perspectives of practitioners and, in particular, with their understandings about trust. In this paper, we address these absences through looking at a critical technology of governmentality--child immunisation. We do this, first, by examining theoretical links between risk, trust and knowledge in relation to the governance of health. We then draw on findings from our secondary analysis of qualitative data based on interviews with key actors in primary care in Scotland charged with the delivery of a particular immunisation--the MMR vaccine. While many practitioners, like parents, have typically perceived immunisation--along with other public health initiatives-as obligatory and as part of 'good citizenship', we argue that in relation to MMR and in the context of clinical governance, practitioners are having to engage in complex negotiations about knowledge and trust--negotiations that are at the heart of governing health at a distance. PMID- 16046043 TI - Postnatal development of rat pups is altered by prenatal methamphetamine exposure. AB - There are studies showing that drug abuse during pregnancy may have impairing effect on progeny of drug-abusing mothers. Methamphetamine (MA) is one of the most common illicit drugs throughout the world. The purpose of the present study was to assess the effect of prenatal MA exposure on postnatal development of rat pups before the time of separation from their mothers. Female rats were injected with MA (5 mg/kg daily) for the duration of their pregnancy. Pups were then tested throughout the lactation period. They were weighed daily and the ano genital distance was measured on postnatal day (PD) 1. Development of postural motor reaction was tested by righting reflex on surface between PD 1 and 12, and righting reflex in mid-air after PD 12 until successfully accomplished. On PD 15 homing test was examined as a test of pup acute learning. On PD 23 sensory-motor coordination was examined using the rotarod and bar-holding tests. Additionally, the markers of physical maturation, such as eye opening, testes descent in males and vaginal opening in females were also recorded. The birth weight in prenatally MA-exposed pups was lower than in controls or saline-exposed pups regardless of sex. There were no changes induced by prenatal MA exposure in weight gain or in sexual maturation. In righting reflexes, we demonstrated that pups exposed prenatally to MA were slower in righting reflex on surface and that they accomplished the test of righting reflex in mid-air later than controls or saline exposed pups. The performance of homing test was not affected by prenatal drug exposure. The sensory-motor coordination was impaired in prenatally MA-exposed pups when testing in the rotarod test. Also, the number of falls in the bar holding test was higher in MA-exposed pups than in controls. There were no sex differences in any measures. Thus, the present study demonstrated that prenatal MA exposure impairs development of postural motor movements of rat pups during the first 3 weeks after birth, while not affecting physical or sexual maturation. PMID- 16046044 TI - Hexachlorocyclohexane differentially alters the antioxidant status of the brain regions in rat. AB - Hexachlorocyclohexane (HCH), a highly persistent organochlorine insecticide, is neurotoxic at acute doses and causes degenerative effects on chronic exposure. HCH has been reported to induce oxidative stress in cells and tissues. Mammalian brain is sensitive to oxidative stress which is implicated in neurodegenerative diseases. Effect of HCH on the brain regions, cortex, cerebellum, midbrain and brainstem, has been investigated by studying the response of antioxidant enzymes in rats treated orally with HCH at 25 and 100mg/kg b.w. for 2 weeks. Lipid peroxidation and glutathione depletion was seen in all the brain regions of HCH treated rats. The brain regions showed distinct variation in the antioxidant enzyme activities. Activities of glutathione peroxidase, glutathione reductase, glutathione-S-transferase and catalase were markedly induced whereas superoxide dismutase was inhibited at higher dose in all the brain regions. Marked induction and inhibition of the antioxidant enzymes, especially in the cortex and to varying degrees in other brain regions, was seen in HCH treated rats. These biochemical changes suggest vulnerability to oxidative stress in the brain is region-specific. Whether these changes are adaptive or compromise the capacity of the brain to deal with the HCH-induced oxidative stress that could lead to degenerative neurotoxic manifestations remain to be understood. PMID- 16046045 TI - Plasmalogen metabolism-related enzymes in rat brain during aging: influence of n 3 fatty acid intake. AB - Plasmalogens (Pls) are phospholipids containing a vinyl-ether bond at the sn-1 position of the glycerol backbone. They represent between 1/2 and 2/3 of the ethanolamine phospholipids in the brain. During aging, the Pls content in human brain falls down. However, the role of Pls metabolism-related enzymes in the regulation of Pls levels remains to be determined. Dihydroxyacetone phosphate acyltransferase (DHAP-AT) is the enzyme involved in the first step of Pls biosynthesis. In the brain, a phospholipase A2, which selectively acts on Pls, has been isolated (Pls-PLA2s). In this work, we aimed to evaluate the impact of DHAP-AT (a key enzyme of Pls biosynthesis) and Pls-PLA2 (a specific Pls degradation enzyme) on the evolution of Pls content in the rat brain during aging. The influence of n-3 fatty acid intake was also evaluated. Littermates from two generations of n-3 deficient rats were fed an equilibrated diet containing either alpha-LNA alone or with two doses of DHA. After weaning, 3, 9 or 21 months of diet, rats were sacrificed. Enzymatic assays were performed, Pls levels were assessed and the sn-2 position of ethanolamine Pls was analyzed. DHAP AT activity significantly increased between weaning and 3 months with a concomitant increase of brain Pls, which reached maximal levels after 9 months. Then, Pls levels and DHAP-AT activity significantly decreased while Pls-PLA2s activity significantly increased. Dietary n-3 fatty acids had no effect on DHAP AT activity and on Pls levels. In conclusion, the increase of brain Pls content in the first part of the life may be related to the high increase of DHAP-AT activity, probably stimulated by DHA. In aged animals, the decrease of Pls levels may mainly be caused to an increase of their degradation by Pls-PLA2. Dietary DHA may not oppose the physiologic aging. PMID- 16046046 TI - A new approach to preventing relapse in opiate addicts: a psychometric evaluation. AB - The present study investigated psychological aspects after different methods of withdrawal treatments in opiate addicts had been conducted. Two pharmacological strategies based on delivering an opioid agonist or antagonist were used for withdrawal in opiate addicts. After detoxification, the antagonist was delivered by a pellet implanted subcutaneously. Four days after the beginning of the treatment several psychological variables such as craving, anxiety, depression, and mood were assessed and compared with data from actively consuming opiate addicts and healthy controls. In addition, 6 and 12 weeks later the relapse rates were assessed. Compared with addicts detoxified and treated with Levomethadone as well as actively heroin consuming addicts, subjects treated with Naltrexone demonstrated significantly higher positive psychological outcome concerning all assessed variables and significantly lower relapse rates. Naltrexone implants prove prevention of relapse during the most vulnerable period after detoxification. Compared with Levomethadone withdrawal, they lead to a significantly better psychological condition in patients. PMID- 16046047 TI - Concentrations of interleukin (IL)-1alpha, IL-1 soluble receptor type II (IL-1 sRII) and IL-1 receptor antagonist (IL-1 Ra) in the peritoneal fluid and serum of infertile women with endometriosis. AB - OBJECTIVE: Endometriosis is an immune system-related gynaecological disease, characterised by an increase in number and activation of peritoneal macrophages. One of macrophage-derived factors is interleukin (IL)-1. The effects of IL-1 are inhibited by IL-1 receptor type II (IL-1 RII), soluble forms of IL-1 RII (IL-1 sRII) and IL-1 receptor antagonist (IL-1 Ra). The aim of our work was to study the IL-1alpha, IL-1 sRII and IL-1 Ra levels in the peritoneal fluid (PF) and serum of women with endometriosis in relation to stage of disease. STUDY DESIGN: Concentrations of IL-1alpha, IL-1 sRII and IL-1 Ra were measured by ELISA assay in the PF and serum of 58 women; 43 with and 15 without endometriosis (control group). RESULTS: Elevated PF and serum IL-1alpha and IL-1 Ra levels in the women with endometriosis in comparison with the control group were observed. IL-1 sRII levels in PF and serum were higher in the controls than in the women with endometriosis. Concentrations of IL-1alpha and IL-1 sRII were higher in advanced endometriosis, but higher IL-1 Ra was observed in the early stage of the disease. CONCLUSION: Impairment of regulation IL-1 activity in the peritoneal fluid and serum of women with endometriosis may play an important role in the pathogenesis and development of the disease. PMID- 16046048 TI - Statistical kinetic approach for modeling lifespan. AB - Lifespan regulation through gene expression involves complex biochemical processes. Unfortunately, current mathematical models for treating lifespan data afford little insight into the mechanisms that control longevity. In this work, we demonstrate the use of a novel kinetic model to successfully fit the lifespan curves of the nematode, Caenorhabditis elegans. Our findings show that population aging may be treated analogously to a dispersive chemical process [P.J. Skrdla, R.T. Robertson., J. Phys. Chem. B 109 10611 (2005)]. Much like the Gompertz model, only two fit parameters, alpha and beta, are needed to adequately describe the entire data set for each nematode population. These parameters relate a 'global first-order time constant' and a 'global second-order rate constant', with units of (time) and (time)(-2), respectively. In C. elegans, the increased longevity resulting from DAF-16 (a transcription factor) activity in the intestinal tissue correlates with a larger alpha value and a smaller beta value; the opposite is true for animals with shorter lifespans. A basic physical interpretation of the two parameters is provided. PMID- 16046049 TI - Application of a new type of moving bio-film in aerobic sequencing batch reactor (aerobic-SBR). AB - A moving bio-film (MB), made from the inner tube of used tyres was applied in a conventional-aerobic-SBR for increasing the system efficiency and quality of bio sludge due to good sedimentation (the density of 1.925+/-0.21 g/cm(3)), non biodegradability and re-usability of the media without any regeneration. The total bio-sludge mass of the MB-aerobic-SBR was about 30% higher than that of the conventional-aerobic-SBR resulting in a reduction of the F/M value of the system and amount of suspended bio-sludge waste. The amount of suspended bio-sludge waste, SVI and SRT of the MB-aerobic-SBR under a low organic loading of 80+/-9.3g BOD(5)/m(3)-d were 1,485+/-146 mg/d, 51+/-3.7 ml/g and 10.1+/-5.1 days, respectively while they were 1,800+/-152 mg/d, 69+/-4.0 ml/g and 8.3+/-5.3 days, respectively in the conventional-aerobic-SBR. The BOD(5), TKN and TP removal efficiencies of the MB-aerobic-SBR were about 1-2, 2-3 and 10-12% higher, respectively, than that of the conventional-aerobic-SBR. Also, the BOD(5) and COD removal efficiencies of the MB-aerobic-SBR were higher than 95% even when the system was operated with synthetic wastewater containing 800 mg/l BOD(5) under a very low HRT of 1.5 days (organic loading of 528+/-50.8 g BOD(5)/m(3)-d). The effluent BOD(5), COD, total kjeldahl nitrogen, total phosphorus and suspended solids of the MB-aerobic-SBR under a high organic loading of 528+/-50.8 g BOD(5)/m(3)-d were 45+/-5.1, 37+/-3.6, 4.1+/-1.0, 1.5+/-0.80 and 41+/-2mg/l, respectively. PMID- 16046050 TI - In vitro tumoral progression of human bladder carcinoma: role for TGFbeta. AB - OBJECTIVE: Investigating whether extracellular factors are possible actors in tumoral progression in bladder carcinoma. METHODS: RT112/G2 bladder tumour cells were grown in presence of TGFbeta and analysed by immunological and cDNA microarray techniques. RESULTS: TGFbeta inhibited cell proliferation, reduced TNFalpha- and IFNgamma-induced apoptosis by decreasing TNFalpha-RI and IFNgamma-R antigen expression. It also inhibited cleaved caspase 8 and 9 expression, decreased E-cadherin, and increased BclxL and cyclooxygenase-2 expression. The cDNA microarray approach showed that TGFbeta up-regulated the expression of genes with defined roles in tumoral progression sometimes associated with poor outcome in bladder cancer. CONCLUSION: These results suggest that a part of the bladder tumoral progression process may be related to the action of exogenous TGFbeta confirming the possible role for the microenvironment. PMID- 16046051 TI - Does the tertiary Gleason pattern influence the PSA progression-free interval after retropubic radical prostatectomy for organ-confined prostate cancer? AB - INTRODUCTION: The Gleason sum is an important prognostic parameter for patients treated with radical prostatectomy for localized prostate cancer. However, frequently more than two predominant Gleason patterns are present in one specimen. In this study we investigated the prognostic significance of tertiary Gleason patterns in radical prostatectomy specimens. PATIENTS AND METHODS: Between 1994 and 2001, 277 patients underwent radical retropubic prostatectomy (RRP) for clinically localised prostate cancer in our institute. We collected information on Gleason score and cancer volume (CV) for all tumour localizations, clinical and pathological stage, seminal vesicle invasion (SVI) and extra capsular extension (ECE). In case one pattern was seen in more than 95% of the tumour, this pattern was used both for the primary and secondary Gleason pattern, and any other pattern (actually the secondary pattern) was called tertiary. Charts were examined retrospectively for clinical follow up. PSA progression was defined as two subsequent rising PSA measurements above 0.10 ng/ml. Kaplan-Meier time to PSA progression was compared between patients with and without a tertiary pattern. RESULTS: Overall, of the 223 patients, 106 (48%) were found to have a tertiary pattern, which on average, was 7% of the total tumour volume. Patients with a tertiary pattern had a 5-year risk of PSA progression of 37.3% versus 12.6% in case no tertiary Gleason pattern was present (log rank p=0.0002). There was no prognostic difference between patients with a higher-grade tertiary pattern as compared to those with a lower grade tertiary pattern. CONCLUSIONS: If present, a tertiary Gleason pattern, whether better or worse than the primary or secondary pattern, is an indication for a worse outcome, as indicated by a shorter time to PSA progression. This suggests that tumour multifocality, rather than the presence of a higher-grade tertiary Gleason pattern has prognostic value. PMID- 16046052 TI - EAU guidelines on prostate cancer. AB - OBJECTIVES: The first summary of the European Association of Urology (EAU) guidelines on prostate cancer was published in 2001. These guidelines have been continuously updated since many important changes affecting the clinical management of patients with prostate cancer have occurred over the past years. The aim of this paper is to present a summary of the 2005 update of the EAU guidelines on prostate cancer. METHODS: A literature review of the new data has been performed by the working panel. The guidelines have been updated and level of evidence/grade of recommendation added to the text. This enables readers to better understand the quality of the data forming the basis of the recommendations. RESULTS: A full version is available at the EAU Office or at . Systemic prostate biopsies under ultrasound guidance is the preferred diagnostic method and the use of periprostatic injection of a local anaesthetic can significantly reduce pain/discomfort associated with the procedure. Active treatment (surgery or radiation) is mostly recommended for patients with localized disease and a long life expectancy with radical prostatectomy being the only treatment evaluated in a randomized controlled trial. Follow-up is at large based on prostate specific antigen (PSA) and a disease-specific history with imaging only indicated when symptoms occur. Cytotoxic therapy has become an option for selected patients with hormone refractory prostate cancer. CONCLUSION: The knowledge in the field of prostate cancer is rapidly changing. These EAU guidelines on prostate cancer summarize the most recent findings and put them into clinical practice. PMID- 16046053 TI - Minimally invasive treatment of renal cell carcinoma: comparison of 4 different monopolar radiofrequency devices. AB - OBJECTIVES: Radiofrequency Ablation is an investigational treatment option for RCC. The aim of our study was to test the ablation algorithms of four different RF systems in a standardized ex vivo perfused porcine kidney model. MATERIALS AND METHODS: A multiline monopolar dry electrode (impedance-based system), a multiline monopolar dry electrode (temperature-based system), a single monopolar wet electrode (impedance-based system) and a single monopolar dry, internally cooled electrode (impedance-based system) were selected. RF energy was applied at different treatment parameters (power with and without control, tissue temperature, saline enhancement) for 1, 3, 5 and 9 minutes in healthy perfused ex vivo porcine tissue. Each treatment parameter was repeated 5 times. Maximum vertical, long-axis and short-axis diameters of the macroscopic lesion were measured and lesion volumes/ shapes were calculated. RESULTS: Lesion volumes increased significantly with the pre-selected tissue temperature and saline enhancement. Saline enhancement created larger, but irregular shaped lesions. The impedance-based system created lesion volumes that were predictable by treatment time and generator power. Lesions were unpredictable when uncontrolled generator power was applied. The created lesion shape was dependent on the selected electrode configuration. CONCLUSIONS: The currently available monopolar RFA systems offer different specific technical features to control tissue ablation. Detailed knowledge of the specific characteristics of each RF system is necessary to provide a higher chance of successful clinical outcome by complete and reliable ablation. PMID- 16046054 TI - Detection of mRNA levels for the estrogen alpha, estrogen beta and androgen nuclear receptor genes in archival breast cancer tissue. AB - Previous studies in our laboratory have shown association of nuclear receptor expression and histological breast cancer grade. To further investigate these findings, it was the objective of this study to determine if expression levels of the estrogen alpha, estrogen beta and androgen nuclear receptor genes varied in different breast cancer grades. RNA extracted from paraffin embedded archival breast tumour tissue was converted into cDNA and cDNA underwent PCR to enable quantitation of mRNA expression. Expression data was normalised against the 18S ribosomal gene multiplex and analysed using ANOVA. Analysis indicated a significant alteration of expression for the androgen receptor in different cancer grades (P=0.014), as well as in tissues that no longer possess estrogen receptor alpha proteins (P=0.025). However, expression of estrogen receptors alpha and beta did not vary significantly with cancer grade (P=0.057 and 0.622, respectively). Also, the expression of estrogen receptor alpha or beta did not change, regardless of the presence of estrogen receptor alpha protein in the tissue (P=0.794 and 0.716, respectively). Post-hoc tests indicate that the expression of the androgen receptor is increased in estrogen receptor negative tissue as well as in grade 2 and grade 3 tumours, compared to control tissue. This increased expression in late stage breast tumours may have implications to the treatment of breast tumours, particularly those lacking expression of other nuclear receptor genes. PMID- 16046055 TI - Lack of enhancing effects of degraded lambda-carrageenan on the development of beta-catenin-accumulated crypts in male DBA/2J mice initiated with azoxymethane. AB - Effect of degraded lambda-carrageenan, which induces colitis in rodents, on the development of beta-catenin-accumulated crypts (BCAC) being putative precancer lesions of colon cancer was investigated in male DBA/2J mice initiated with azoxymethane (AOM). In a preliminary experiment, male DBA/2J mice among seven different strains (A/J, BALB/c, C3H/HeN, C57BL/6J, CBA/N, DBA/1J, and DBA/2J) of male mice were most sensitive to degraded lambda-carrageenan. Therefore, male DBA/2J mice were intraperitonially injected AOM (10 mg/kg body weight), and then 2% degraded lambda-carrageenan in drinking water for one or two weeks, starting one week after dosing of AOM. Thereafter animals were no further treated up to week 26. At week 26, the frequency of BCAC in the colonic mucosa was 12.50+/-2.46 in the AOM alone group, 11.30+/-3.50 in the AOM/degraded lambda-carrageenan (for one week) group, and 11.60+/-2.27 in the AOM/degraded lambda-carrageenan (for two weeks) group. The findings suggest that degraded lambda-carrageenan treatment for one or two weeks did not affect the occurrence of BCAC. Our results may indicate no enhancing or promoting effects of degraded lambda-carrageenan on colon carcinogenesis in mice initiated with AOM. PMID- 16046056 TI - Tissue-specificity of prostate specific antigens: comparative analysis of transcript levels in prostate and non-prostatic tissues. AB - Activation of immune defense mechanisms against tumor antigens appears to be a promising therapeutic option for advanced prostate cancer (PCa). Specific immunotherapy critically depends on target antigens that are selectively expressed in the tumorous and optional in the normal prostate tissue in sufficient amounts. Although several prostate antigens have been described and some have already been used in clinical trials, a detailed comparative evaluation of their tissue-specificity and expression levels is still lacking. We determined the transcript levels of eight prostate targets (PSA, PAP, PSCA, PSGR, Prostein, PSMA, AIbZIP, trp-p8) in 16 different tissues by quantitative PCR and calculated a tissue-specificity index (TSI) for each molecule. Besides a preferential expression in prostate for all targets, striking differences in the expression levels and TSI were revealed which may be important for the selection of appropriate antigens for immunotherapy of PCa. PMID- 16046057 TI - Correlation of reversion-inducing cysteine-rich protein with kazal motifs (RECK) and extracellular matrix metalloproteinase inducer (EMMPRIN), with MMP-2, MMP-9, and survival in colorectal cancer. AB - mRNA, and latent and active levels MMP-2 and -9 were higher in tumor tissue compared to normal tissue from 63 patients with colorectal cancer, whereas RECK and EMMPRIN levels were lower. Correlations between mRNA, latent, and active MMP were particular high for MMP-2 in tumor tissue (R(s)=0.6-0.8, P<0.001). For active MMP-2, but not for MMP-9, a significant negative partial correlation (R(p)=-0.440, P<0.001) for RECK was found in tumor tissue, which was confirmed by linear regression analysis. In exploratory survival analyses we found that in patients with localized disease the RECK level in normal or tumor tissue had a significant (P=0.017) association with overall survival. PMID- 16046058 TI - Cathepsin D: newly discovered functions of a long-standing aspartic protease in cancer and apoptosis. AB - The lysosomal aspartic protease cathepsin D (cath-D) is over-expressed and hyper secreted by epithelial breast cancer cells. This protease is an independent marker of poor prognosis in breast cancer being correlated with the incidence of clinical metastasis. Cath-D over-expression stimulates tumorigenicity and metastasis. Indeed it plays an essential role in the multiple steps of tumor progression, in stimulating cancer cell proliferation, fibroblast outgrowth and angiogenesis, as well as in inhibiting tumor apoptosis. A mutated cath-D devoid of catalytic activity still proved mitogenic for cancer, endothelial and fibroblastic cells, suggesting an extra-cellular mode of action of cath-D involving a triggering, either directly or indirectly, of an as yet unidentified cell surface receptor. Cath-D is also a key mediator of induced-apoptosis and its proteolytic activity has been involved generally in this event. During apoptosis, mature lysosomal cath-D is translocated to the cytosol. Since cath-D is one of the lysosomal enzymes which requires a more acidic pH to be proteolytically active relative to the cysteine lysosomal enzymes, such as cath-B and -L, it is open to question whether cytosolic cath-D might be able to cleave substrate(s) implicated in the apoptotic cascade. This review summarises our current knowledge on cath-D action in cancer progression and metastasis, as well as its dual function in apoptosis. PMID- 16046059 TI - Sonochemical treatment of fly ash for dye removal from wastewater. AB - Fly ash samples modified by NaOH solution and sonochemical treatment were tested for a basic dye (methylene blue) adsorption in aqueous solution. It is found that sonochemical treatment of fly ash can significantly increase the adsorption capacity depending on the concentration of NaOH and treatment time. The untreated FA and the sonochemically treated sample exhibits adsorption capacity at 6 x 10( 6)mol/g and 1.2 x 10(-5)mol/g at 30 degrees C, respectively. The adsorption tests show that solution pH and adsorption temperature also influence the adsorption behaviour. The adsorption isotherms can be fitted by Langmuir and Freudlich models, while the two-site Langmuir heterogeneous model will present the best result. PMID- 16046060 TI - A rating system for determination of hazardous wastes. AB - Although hazardous waste lists and their classification methodologies are nearly the same in most of the countries, there are some gaps and subjectiveness in determining the waste as hazardous waste. A rating system for the determination of waste as a hazardous waste is presented in this study which aims to overcome the problems resulted from the existing methodologies. Overall rating value (ORV) calculates and quantifies the waste as regular, non-regular or hazardous waste in an "hourglass" scale. "ORV" as a cumulative-linear formulation in proposed model consists of components such as ecological effects of the waste (Ee) in terms of four main hazard criteria: ignitability, reactivity, corrosivity and toxicity; combined potential risk (CPR) including carcinogenic effect, toxic, infectious and persistence characteristics; existing lists and their methodology (L) and decision factor (D) to separate regular and non-regular waste. Physical form (f) and quantity (Q) of the waste are considered as factors of these components. Seventeen waste samples from different sources are evaluated to demonstrate the simulation of the proposed model by using "hourglass" scale. The major benefit of the presented rating system is to ease the works of decision makers in managing the wastes. PMID- 16046061 TI - Prediction of optimum sorption isotherm: comparison of linear and non-linear method. AB - Equilibrium parameters for Bismarck brown onto rice husk were estimated by linear least square and a trial and error non-linear method using Freundlich, Langmuir and Redlich-Peterson isotherms. A comparison between linear and non-linear method of estimating the isotherm parameters was reported. The best fitting isotherm was Langmuir isotherm and Redlich-Peterson isotherm equation. The results show that non-linear method could be a better way to obtain the parameters. Redlich Peterson isotherm is a special case of Langmuir isotherm when the Redlich Peterson isotherm constant g was unity. PMID- 16046062 TI - The effect of pH, temperature and concentration on electrooxidation of phenol. AB - The electrochemical oxidation of phenol on Pt electrode has been investigated in 1M NaOH and 0.5M H2SO4 solutions at different temperature and different phenol concentration. A simplified mechanism for the electrochemical oxidation of phenol is proposed which includes a selective oxidation parallel with PtO(x) formation. Activation energy, which was obtained from the experimental data, was almost same in studied mediums. This means that electrooxidation mechanism for phenol must be same. Other thermodynamic values such as DeltaS, DeltaH were calculated from the experimental data. PMID- 16046063 TI - Natural attenuation of MTBE at two petroleum-hydrocarbon spill sites. AB - Methyl tert-butyl ether (MTBE) has been used as a gasoline additive to improve the combustion efficiency and to replace lead since 1978. Because it is widely used and it has been disposed inappropriately, MTBE has become a prevalent groundwater contaminant worldwide. In this study, two petroleum-hydrocarbon contaminated sites (Sites A and B) were selected to evaluate the occurrence and effectiveness of natural attenuation of MTBE at these two sites. Field investigation results indicate that the natural attenuation mechanisms of MTBE at both sites were occurring with the first-order attenuation rates of 0.0021 and 0.0048 1day(-1) at Sites A and B, respectively. Results also reveal that the intrinsic biodegradation pattern was the most important mechanism among the natural attenuation processes at both sites. Results from BIOSCREEN simulation suggest that biodegradation was responsible for 78 and 59% of MTBE mass reduction at Sites A and B, respectively. Investigation results show that MTBE plume at Site B could be effectively controlled via natural attenuation processes. However, MTBE plume at Site A has migrated to a farther downgradient area and passed the boundary line of the site. Thus, more active groundwater remedial technologies should be applied at Site A to protect the downgradient environment. Results from this study suggest that natural attenuation might be feasible to be used as a remedial option for the remediation of MTBE-contaminated site on the premise that (1) detailed site characterization has been conducted and (2) the occurrence and effectiveness of natural attenuation processes have been confirmed. PMID- 16046064 TI - Brain activation of spider phobics towards disorder-relevant, generally disgust- and fear-inducing pictures. AB - The present functional magnetic resonance imaging study investigated the fear and disgust reactivity of patients suffering from spider phobia. Ten phobics and 13 control subjects were scanned while viewing alternating blocks of phobia relevant, generally fear-inducing, disgust-inducing and affectively neutral pictures. The patient group rated the spider pictures as being more disgust and fear evoking than the control group, and showed greater activation of the visual association cortex, the amygdalae, the right dorsolateral prefrontal cortex and the right hippocampus. Specific phobia-related activation occurred in the supplementary motor area. The patients also showed greater amygdala activation during the presentation of generally disgust- and fear-inducing pictures. This points to an elevated sensitivity to repulsive and threatening stimuli in spider phobics and implicates the amygdala as a crucial neural substrate. PMID- 16046065 TI - The controlled differentiation of human neural stem cells into TH-immunoreactive (ir) neurons in vitro. AB - The expansion of human neural stem cells in vitro might overcome the poor donor supply of human fetal neural tissue in transplantation for Parkinson's disease. However, the differentiation of human neural stem cells into dopaminergic neurons has proven difficult. In the present study, we investigated the effects of cytokines, trophic factors of developmental striatum and Ginkgolide on differentiation of human neural stem cells (hNSCs) into TH-ir neurons. The immunoreactivity to tyrosine hydroxylase (TH), a distinctive marker for dopamine neurons was used to assess dopaminergic neuronal phenotype. We demonstrate that human neural stem cells expanded in vitro can efficiently differentiate into TH ir neurons by induction. These stem cells might serve as a continuous, on-demand source of cells for therapeutic transplantation in patients with Parkinson's disease. PMID- 16046066 TI - Language lateralization development in children with autism: insights from the late field magnetoencephalogram. AB - Left hemisphere dominance represents the typical language lateralization profile for the majority of neurologically healthy, right-handed individuals. We investigated hemispheric dominance for language in language-impaired children with autism and typically developing controls to investigate the hypothesis that atypical functional specialization for language represents one component of developmental language impairment in autism. Late field magnetoencephalography (MEG) recordings were used to calculate a hemispheric Lateralization Index from the neuromagnetic activity evoked by passive auditory presentation of vowel stimuli. Results indicate that children with autism and typically developing children follow opposite maturational trajectories in language lateralization; while leftward lateralization (i.e. left hemisphere dominance) emerged from bilaterally symmetric neuronal activation as age increased in our sample of typically developing children, rightward lateralization emerged from bilaterally symmetric activity as age increased in our sample of children with autism. PMID- 16046067 TI - A practical method to detect and correct for lens distortion in the TEM. AB - A practical, offline method for experimental detection and correction for projector lens distortion in the transmission electron microscope (TEM) operating in high-resolution (HR) and selected area electron diffraction (SAED) modes is described. Typical TEM works show that, in the simplest case, the distortion transforms on the recording device, which would be a circle into an ellipse. The first goal of the procedure described here is to determine the elongation and orientation of the ellipse. The second goal is to correct for the distortion using an ordinary graphic program. The same experimental data set may also be used to determine the actual microscope magnification and the rotation between SAED patterns and HR images. The procedure may be helpful in several quantitative applications of electron diffraction and HR imaging, for instance while performing accurate lattice parameter determination, or while determining possible metrical deviations (cell edges and angles) from a given symmetry. PMID- 16046068 TI - Determination of the Fe content of embedded Cu-rich particles in ferritic alloys using energy-filtered TEM. AB - A novel technique for the quantification of the iron content of copper precipitates in ferritic steels is presented. Energy-filtered (EF) imaging has been used to extract elemental maps with high spatial resolution. These maps contain enough information to attempt the quantification of the signal produced by the precipitates when either a line profile is measured across them or the whole image signal is integrated. Assumptions such as sphericity of the precipitates and composition variations are discussed. Special attention to the assessment of drift on the information extracted from EF images has been taken. Minimum detectability and optimum acquisition conditions are discussed. PMID- 16046069 TI - Wasting and mortality in beef cattle parasitized by Eurytrema coelomaticum in the State of Parana, southern Brazil. AB - A sporadic wasting syndrome affecting beef cattle herds parasitized by Eurytrema coelomaticum is described in the State of Parana, southern Brazil. The disease was characterized by progressive weight loss, poor body condition despite plenty of good quality forage available, and death. Annual losses ranged from 1 to 3%. The clinical course of the disease varied from 2 to 10 months after the onset of the first clinical signs. At necropsy, one of the three distinct following patterns of lesions were observed: the pancreas was of normal size and color; small, shrunken, white and markedly and diffusely firm (fibrosis); or slightly enlarged and dark with a shriveled capsular surface. Myriads of leaf-shaped trematodes of the genus Eurytrema were packed inside multiple dilated ducts with thickened, whitish fibrous walls, or the flukes were embedded in the remaining pancreatic parenchyma. Microscopic findings included extensive loss of the pancreatic parenchyma with replacement fibrosis, intralesional flukes and eggs, and ductal hyperplasia. Inflammatory reaction varied from absent to severe with the presence of a granulomatous reaction around the trematode eggs. One affected animal had high plasma amylase concentration (1580U/L) suggesting exocrine pancreatic insufficiency. Glucose blood levels were not significantly increased. We suggest that diffuse and marked fibrosis of the pancreas induced by the presence of large amounts of pancreatic flukes is the cause of the sporadic cases of chronic wasting and death of cattle in this geographic area. PMID- 16046070 TI - A structural model of a seven-transmembrane helix receptor: the Duffy antigen/receptor for chemokine (DARC). AB - The Duffy antigen/receptor for chemokine (DARC) is an erythrocyte receptor for malaria parasites (Plasmodium vivax and Plasmodium knowlesi) and for chemokines. In contrast to other chemokine receptors, DARC is a promiscuous receptor that binds chemokines of both CC and CXC classes. The four extracellular domains (ECDs) of DARC are essential for its interaction with chemokines, whilst the first (ECD1) is sufficient for the interaction with malaria erythrocyte-binding protein. In this study, we elaborate and analyze structural models of the DARC. The construction of the 3D models is based on a comparative modeling process and on the use of many procedures to predict transmembrane segments and to detect far homologous proteins with known structures. Threading, ab initio, secondary structure and Protein Blocks approaches are used to build a very large number of models. The conformational exploration of the ECDs is performed with simulated annealing. The second and fourth ECDs are strongly constrained. On the contrary, the ECD1 is highly flexible, but seems composed of three consecutive regions: a small beta-sheet, a linker region and a structured loop. The chosen structural models encompass most of the biochemical features and reflect the known experimental data. They may be used to analyze functional interaction properties. PMID- 16046071 TI - Inhibition of several protein phosphatases by a non-covalently interacting microcystin and a novel cyanobacterial peptide, nostocyclin. AB - Microcystins produced by cyanobacterial 'blooms' in reservoirs and lakes pose significant public health problems because they are highly toxic due to potent inhibition of protein serine/threonine phosphatases in the PPP family. A dehydrobutyrine (Dhb)-containing microcystin variant [Asp3, ADMAdda5, Dhb7]microcystin-HtyR isolated from Nostoc sp. was found to potently inhibit PP1, PP2A, PPP4 and PPP5 with IC50 values similar to those of microcystin-LR. However, in contrast to microcystin-LR, which forms a covalent bond with a cysteine residue in these protein phosphatases, Asp,ADMAdda,Dhb-microcystin-HtyR did not form any covalent interaction with PP2A. Since the LD50 for Asp,ADMAdda,Dhb microcystin-HtyR was 100 microg kg(-1) compared to 50 microg kg(-1) for microcystin-LR, the data indicate that the non-covalent inhibition of protein phosphatases accounts for most of the harmful effects of microcystins in vivo. A 3-amino-6-hydroxy-2-piperidone containing cyclic peptide, nostocyclin, also isolated from Nostoc sp., was non-toxic and exhibited more than 500-fold less inhibitory potency towards PP1, PP2A, PPP4 and PPP5, consistent with the conclusion that potent inhibition of one or more these protein phosphatases underlies the toxicity of microcystins, both lacking and containing Dhb. PMID- 16046072 TI - Dynamic joint-aligned PIP and DIP corrective-flexion/extension orthosis for post burn finger contractures. PMID- 16046073 TI - An unusual cause of burns due to cupping: complication of a folk medicine remedy. PMID- 16046074 TI - Rhabdomyolysis caused by hot air sauna burn. AB - Sauna-related burns are rare, even in Finland where sauna bathing is a popular leisure pastime. Burns induced by hot air are even more rare and constitute a very small subgroup of all sauna burns. Hot air burns are characterised by a combination of full thickness skin damage with deep tissue destruction. We report here on six consecutive patients suffering from hot air sauna burns with rhabdomyolysis. All six patients were middle-aged, the majority of them men. Acute excessive consumption of alcohol exacerbated by a hot environment resulted in dehydration and loss of consciousness. Immobility and prolonged exposure to hot, dry air resulted in third degree regional burns with 5-32% TBSA. Rhabdomyolysis was evident on admission. The laboratory values of plasma creatine kinase (P-CK), plasma myoglobin (PM), blood pH, and serum potassium (S-K) during the first five days were evaluated. Aggressive fluid management and correction of acidosis and myoglobinuria were started on admission. Surgical management consisted of early, aggressive excision at fascial level, in some patients involving sacrifice of the upper layers of muscle. Even so, mortality in this small series was 50%. The best indicator of poor prognosis was a highly elevated CK value on the second post-injury day. PMID- 16046075 TI - An unusual burn with varnish: a case report. PMID- 16046076 TI - Salivary alpha amylase as marker for adrenergic activity during stress: effect of betablockade. AB - Free salivary cortisol is an established non-invasive marker of hypothalamus pituitary adrenal (HPA) axis activity. In contrast, such a well-characterized salivary marker for activity of the sympatho-adrenal medullar (SAM) system is still missing. As one potential candidate salivary alpha amylase (sAA) has been suggested. In humans increases in sAA levels have been observed in response to physiological and psychological stress. The present study aimed at exploring the effects of a pharmacological manipulation (betablockade) on sAA in the context of a stressful fMRI experiment on emotional information processing. Thirty young healthy subjects participated in a double blind group comparison study and received 80 mg of the betablocker (BB) propranolol or a placebo (PL). Salivary samples were obtained before and 90 min (pre-scan) and 135 min (post-scan) after drug application. In addition heart rate and blood pressure were assessed. During rest a significant drug by time interaction was observed, lowering sAA levels as well as heart rate and systolic blood pressure in the betablocker treatment group. During the scanning procedure, in which participants were confronted with highly negative emotional pictures, the significant increase in sAA levels in the PL group compared to the BB group persisted. No additional change was noticed in heart rate or blood pressure during scanning in the PL or BB group. The current pharmacological study in the human provides direct evidence for the sensitivity of sAA to changes in adrenergic activation, specifically in reaction to psychological stress. PMID- 16046077 TI - Social stress, coping strategies and tumor development in male mice: behavioral, neuroendocrine and immunological implications. AB - The relationships between acute social stress, immunological alterations and the development of pulmonary metastases of B16F10 melanoma were analyzed. In particular, the effects of different behavioral coping strategies on the development of the metastases were studied. Tumor bearing and tumor non-bearing mice were subjected for 24h to a sensory contact social stress model. This included two 5 min sessions of direct social interaction with their resident cagemates (which had been selected for consistent levels of aggression). The subjects' behavior was videotaped and assessed. Corticosterone, IL-2, IL-12 and splenic cell proliferation responses to Con-A were determined 1h and 3 days post stress. Lung metastatic foci numbers were determined 21 days after inoculation (15 days post-stress). Social stress increased the number of pulmonary metastases and the serum level of corticosterone but decreased the splenic proliferative capacity. No direct relationship could be established between the development of the metastases and the assayed interleukin response. A combination of cluster and discriminant analyses established that there were three types of coping strategies. Subjects engaging in a strategy characterized by an absence of attack, low non-social exploration levels and high levels of defense, subordination and avoidance, developed most pulmonary metastases. Social stress effects on tumor development appear to depend on the subject's coping strategy in such situations (although one cannot rule out the possibility that differences in the development of the disease per se are responsible for the different behavioral patterns observed). PMID- 16046078 TI - Peer group reputation and smoking and alcohol consumption in early adolescence. AB - Research has shown that substance use might serve beneficial, developmental functions for adolescents, in particular concerning young people's functioning in friendships and intimate relationships. Nonetheless, a major problem in these studies is that information on social functioning is based on adolescent's self reports, which undoubtedly might lead to an overestimation of the social functions of substance use. In the present study, we collected data of 3361 early and mid-adolescents at secondary schools in the Netherlands. Information on individual student's social behavior and psychological traits were gathered by using sociometric measures. All respondents were asked to evaluate all classmates on measures, such as sociability, self-confidence, achievement, withdrawal, and aggression. Person- and variable-centered analyses clearly showed that highest levels of smoking and drinking were found in adolescents who score high on sociability and self-confidence, and relatively low on aggression inattentiveness, achievement-withdrawal, and emotionality-nervousness. This suggests that beneficial functions of substance use are not only in the eyes of the beholder, at least not in that of the individual drinker or smoker. PMID- 16046079 TI - Nanobacteria and intervertebral disc degeneration. PMID- 16046080 TI - Stability and instability in alcohol diagnosis from ages 18 to 21 and ages 21 to 25 years. AB - BACKGROUND: Only in recent years have longitudinal studies of adolescents diagnosed alcohol use disorders and these have not distinguished between abuse and dependence. This study describes the course of disorder from age 18 to age 25 for abuse and dependence and investigates the extent to which continuities in disorder can be explained by background factors. METHODS: A birth cohort of 1,265 individuals from Christchurch, New Zealand, followed annually to age 16 years then at 18, 21 and 25 years (1,003 at age 25). DSM-IV diagnoses were made from reports of alcohol symptoms at 18, 21 and 25 years. RESULTS: The most stable diagnosis was that of no diagnosis, with 83-91% staying the same from one interview to the next. There were high rates of remission to no disorder; 57-75% for those with initial abuse and 50-54% of those with initial dependence. Nonetheless prior diagnosis was a strong predictor of subsequent diagnosis (ORs of 3.7-27.6). Adjustment for background risk factors reduced these odds ratios but all remained significant and substantial (minimum 2.6). CONCLUSIONS: The dual finding of substantial discontinuity and substantial continuity indicates that both public health and treatment interventions are warranted. PMID- 16046081 TI - Antiangiogenic activities of polysaccharides isolated from medicinal fungi. AB - Extracted polysaccharides from medicinal fungi, including Antrodia cinnamomea, Antrodia malicola, Antrodia xantha, Antrodiella liebmannii, Agaricus murrill, and Rigidoporus ulmarius, were investigated for their effects on vascular endothelial growth factor (VEGF)-induced tube formation in endothelial cells (ECs). Chemical analysis revealed that myo-inositol, sorbitol, fucose, galactosamine, glucosamine, galactose, glucose, and mannose were the neutral sugars in these polysaccharides. These fungal polysaccharides showed no toxicity to ECs. For the inhibition of endothelial tube formation, extracted polysaccharides from A. xantha and R. ulmarius were shown to produce greater inhibition compared to those from other fungi. Fucose, glucose and mannose were the predominant monosaccharides from these two fungi. These results suggest that monosaccharides may play a role in the inhibitory effect of these fungi on endothelial tube formation. In contrast to the inhibition on tube formation from polysaccharides of A. cinnamomea and A. malicola, polysaccharides from A. xantha and R. ulmarius, with molecular weight between 2693-2876 and 304-325 kDa, were critical for this inhibitory activity. Our results show that polysaccharides isolated from A. xantha and R. ulmarius provide greater antiangiogenesis than those from commercialized A. murrill (Brazilian mushroom) and A. cinnamomea. These studies provide a basis for the potential development of these polysaccharides for antiangiogenesis usage. PMID- 16046082 TI - Escherichia coli phosphoglucose isomerase can be substituted by members of the PGI family, the PGI/PMI family, and the cPGI family. AB - The Escherichia coli strain Pgi-UdhA, a mutant of the strain MG1655, is deficient in both the pgi gene and the udhA gene and cannot grow on glucose as carbon and energy source. This strain was transformed with different pET-plasmids containing archaeal or bacterial pgi, cpgi or pgi/pmi genes from the three known PGI families (PGI, PGI/PMI, cPGI). Growth could be restored upon plasmid-based expression of pgi, pgi/pmi or cpgi genes indicating that these heterologous proteins can substitute for E. coli PGI. However, complete restoration of the growth rate could not be obtained by any of the PGIs, PGI/PMIs, or cPGIs used. The data indicate that the PGI function of the three PGI families is functionally exchangeable in glycolysis. PMID- 16046083 TI - Characterization of two major groups of diarrheagenic Escherichia coli O26 strains which are globally spread in human patients and domestic animals of different species. AB - Twenty-three Escherichia coli O26 strains from humans, cattle, sheep, pigs and chicken were investigated for virulence markers and for genetic similarity by pulsed field gel electrophoresis and multi locus sequence typing. Two groups of genetically closely related O26 strains were defined. One group is formed by enteropathogenic (EPEC) and enterohemorrhagic (EHEC) E. coli strains, which do not ferment rhamnose and dulcitol and most of these carry a plasmid encoding enterohemolysin. The other group consists of rhamnose and dulcitol fermenting EPEC strains, which carry plasmids encoding alpha-hemolysin. Multiple species of domestic animals were shown to serve as a reservoir for human pathogenic O26 EPEC and EHEC strains. PMID- 16046084 TI - Evolutionary plasticity of methionine biosynthesis. AB - Methionine is an essential cellular constituent, the initiator of protein synthesis and a precursor in many metabolic activities, such as methylation and formylation. Here we investigate the genomic distribution of the methionine biosynthetic pathway and analyze its evolutionary history by reconstructing the phylogeny of its enzymatic components. We demonstrate the evolutionary complexity of methionine synthesis and describe the various mechanisms that have shaped this biosynthetic pathway: gene duplication, functional reassignment, lateral acquisition and gene loss. Lateral gene transfer within and between domains and gene recruitment have played an important role in the evolution of this pathway, especially in its first and third enzymatic steps--homoserine activation and homocysteine methylation. These analyses are also the basis of predictions regarding methionine synthesis in Archaea, where the pathway is yet to be characterized. This study illustrates how diverse molecular solutions can fulfill a conserved function in living beings. PMID- 16046085 TI - Short term heart rate variability in healthy neonates: normative data and physiological observations. AB - BACKGROUND: The measurement of heart rate variability (HRV) reveals information on the functional state of the autonomic nervous system (ANS). Moreover, several diseases are known to be accompanied by a reduction in HRV. Currently, there are no data on HRV within larger samples of healthy neonates. AIM: This study was initiated to establish normative data of HRV parameters and to assess physiological observations in healthy newborn infants. STUDY DESIGN: We performed 10 min recordings of HRV in 80 healthy neonates. We computed time domain parameters. Furthermore power spectrum analysis was calculated by Fourier Transformation in 3 frequency bands. The total power as well as the peak frequencies was estimated for each band. RESULTS: The normative data of each HRV parameter for healthy newborns are presented as mean, standard deviation and the 5th and 95th percentiles. Increasing postnatal age is accompanied by a significant (p<0.05) elevation of HRV parameters. Baroreceptor loop fluctuations were delayed from 0.1 Hz (adults) to 0.07 Hz in healthy newborns. A respiratory peak according to respiratory sinus arrhythmia (RSA) as it is common in adults was not detectable. CONCLUSIONS: Normative data of HRV allows the interpretation of clinical data in neonatal diseases affecting the ANS. Moreover, we observed physiological deviations in healthy newborns in comparison to adults. PMID- 16046086 TI - Is one-wave follicular growth during the estrous cycle a usual phenomenon in water buffaloes (Bubalus bubalis)? AB - The pattern of growth and regression of ovarian follicles was monitored once daily for one complete estrous cycle in eight individual water buffaloes by ultrasonographic scanning of the ovaries for an entire interovulatory interval of normal cycle length. One-wave follicular growth was observed in five animals and two-wave follicular growth in three buffaloes during the estrous cycle. The first follicular wave of a two-wave cycle emerged significantly earlier (P < 0.05) than the emergence of the solitary wave of a one-wave cycle. One- and two-wave cycles differed significantly (P < 0.05) with respect to the mean interovulatory interval (21.0 +/- 0.54 days versus 22.7 +/- 0.33 days) and the mean interestrus interval (20.8 +/- 0.58 days versus 22.3 +/- 0.66 days). The overall linear growth rate of the ovulatory follicle was significantly greater (P < 0.01) in a two-wave cycle compared to that of a one-wave cycle (1.17 +/- 0.33 mm/day versus 0.32 +/- 0.01 mm/day). In a one-wave pattern, the growth profile of the solitary dominant follicle was atypical, showing three distinct phases, i.e. growth phase, regression phase and regrowth phase culminating in ovulation. The level of plasma progesterone steadily increased from day 0 of estrous cycle, attained peak level on day 14 and declined thereafter. A slower growth rate of the dominant follicle was observed in the presence of higher plasma progesterone concentration. The present study shows that one-wave follicular growth is a normal phenomenon in suckled water buffaloes. PMID- 16046087 TI - Novel lipid-based formulations enhancing the in vitro dissolution and permeability characteristics of a poorly water-soluble model drug, piroxicam. AB - Lipid-based delivery systems are becoming increasingly popular as carriers of drugs due to their ability to overcome barriers to oral absorption. The purpose of this study was to prepare novel lipid-based formulations of a model drug, piroxicam (PXCM), a poorly water-soluble non-steroidal anti-inflammatory drug (NSAID) using 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) phospholipid alone, and in combination with polyethylene glycol (PEG 4600). Lipid based drug delivery systems were prepared using conventional methods of preparation and the following aspects were evaluated (1) in vitro dissolution behavior, (2) absorption via Caco-2 cell monolayers and (3) stability of formulations over a 12-month period. In addition, physical characterization studies using differential scanning calorimetry (DSC) were also performed. Formulations of PXCM were prepared using DMPC in the following combinations (A) 1:1 and (B) 2:1 and a mixture of DMPC and PEG 4600 (C) 2:1:1, respectively. Dissolution studies conducted in phosphate buffered saline (PBS, pH 7.4, 37+/-0.5 degrees C) using the USP type II (paddle) dissolution apparatus showed an increase in dissolution rate and extent of the PXCM from all solid dispersion formulations when compared to the control. As such, the rate of drug release was observed to be fastest with formulation (C) showing the greatest increase of over two-fold compared to the control. Release of PXCM from formulations (A) and (B) was intermediate with the latter showing superior dissolution behavior despite containing lower amounts of the carrier lipid than the former. This observation indicates a possible existence of threshold levels for phospholipids carriers beyond which dissolution could be adversely affected. DSC studies further confirmed the dissolution behavior of these formulations demonstrating different levels of amorphous to crystalline nature. Results of HPLC analysis from Caco-2 cell culture studies showed increase in transport of PXCM from all formulations, with formulation (C) showing the maximum increase followed by formulations (B) and (A), when compared to control. The apparent permeability coefficients (Papp) were calculated to be 7.92x10(-6), 9.48x10(-6), 9.2x10(-6) and 5.6x10(-6)cm/s for formulations (A)-(C) and control, respectively. Overall, permeation appeared to improve for all formulations over the control. Stability studies at various temperatures showed all formulations to have good stability for the first 6 months; then a decline in dissolution rates was observed, especially for PEG based lipid carrier systems, attributed to the increase in crystalline content of the solid dispersions upon storage. PMID- 16046088 TI - Anti-HIV-1 activity of propolis in CD4(+) lymphocyte and microglial cell cultures. AB - An urgent need for additional agents to treat human immunodeficiency virus type 1 (HIV-1) infection led us to assess the anti-HIV-1 activity of the natural product propolis in CD4(+) lymphocytes and microglial cell cultures. Propolis inhibited viral expression in a concentration-dependent manner (maximal suppression of 85 and 98% was observed at 66.6 microg/ml propolis in CD4(+) and microglial cell cultures, respectively). Similar anti-HIV-1 activity was observed with propolis samples from several geographic regions. The mechanism of propolis antiviral property in CD4(+) lymphocytes appeared to involve, in part, inhibition of viral entry. While propolis had an additive antiviral effect on the reverse transcriptase inhibitor zidovudine, it had no noticeable effect on the protease inhibitor indinavir. The results of this in vitro study support the need for clinical trials of propolis or one or more of its components in the treatment of HIV-1 infection. PMID- 16046089 TI - New insight in the neuropharmacological activity of Humulus lupulus L. AB - The purpose of the present study was to investigate the effects of Humulus lupulus CO2 extract and its fraction containing alpha-acids on the central nervous system of rats. Both tested substances were able to prolong pentobarbital sleeping time, without affecting the latency to the loss of the righting reflex. This effect was dose-dependent, starting from a minimal dose of 10 mg/kg. Neither the extract nor its alpha-acid fraction affected the locomotor activity in the open field test or exerted an anxiolytic effect in rats submitted to the elevated plus-maze test. Interestingly both compounds reduced the immobility time during the behavioral despair test when administered three times (24, 5 and 1 h) before the test. In conclusion this report shows that Humulus lupulus CO2 extract exerts: (a) a pentobarbital sleep-enhancing property without influencing the motor behavior of rats; (b) an antidepressant activity. The same effects were elicited by the administration of the Humulus lupulus fraction containing alpha acids, which can be considered as the major responsible for the enhanced pentobarbital effect and for the antidepressant property. PMID- 16046090 TI - In vitro cytotoxic, antileishmanial and antifungal activities of ethnopharmacologically selected Gabonese plants. AB - Seventy-seven crude extracts from leaves and stem barks of 15 Gabonese plants used in traditional medicine were evaluated for their cytotoxic, antileishmanial and antifungal activities. Most of the extracts exhibited cytotoxic activities toward human monocytes, and most particularly the hydromethanolic 50% (v/v) fraction of Ganophyllum giganteum leaves (IC(50)=1.3 microg/ml) as well as the methanolic extracts of Polyalthia suaveolens, Dioscorea preussii, Augouardia letestui leaves and Cola lizae stem barks (IC(50)<5 microg/ml). The methanolic extract of Polyalthia suaveolens displayed a strong antiproliferative activity against the promastigote form of Leishmania infantum parasites and presented a good antifungal activity on all the tested strains (IC(50)<1mg/ml). This extract was divided into six fractions: fraction F6 demonstrated a cytotoxic activity stronger than those of the crude extract (IC(50)=0.6 microg/ml), fractions F4 and F5 were devoid of cytotoxicity (IC(50)>100 microg/ml) and displayed interesting antileishmanial activity against the intracellular amastigote form of the parasite (IC(50)=5.6 and 12.4 microg/ml), respectively. However, the antifungal activity observed for the crude extract could not be recovered in the corresponding fractions. PMID- 16046091 TI - DNA profiling of trace DNA recovered from bedding. AB - Trace DNA is often detected on handled items and worn clothing examined in forensic laboratories. In this study, the potential transfer of trace DNA to bedding by normal contact, when an individual sleeps in a bed, is examined. Volunteers slept one night on a new, lower bed sheet in their own bed and one night in a bed foreign to them. Samples from the sheets were collected and analysed by DNA profiling. The results indicate that the DNA profile of an individual can be obtained from bedding after one night of sleeping in a bed. The DNA profile of the owner of the bed could also be detected in the foreign bed experiments. Since mixed DNA profiles can be obtained from trace DNA on bedding, caution should be exercised when drawing conclusions from DNA profiling results obtained from such samples. This transfer may have important repercussions in sexual assault investigations. PMID- 16046092 TI - Correlation of age and ossification of the medial clavicular epiphysis using computed tomography. AB - OBJECT: The aim of this study was the examination of relationship between the age and the ossification of medial epiphysis of the clavicle referred to CT examination. MATERIALS AND METHODS: Concerning the epiphyseal ossification of the clavicle CT's of 100 patients (50 male and 50 female) between 16 and 25 years (10 patients for each year) were analyzed by three viewers. RESULTS: In the legal relevant age segment (16-25a) we saw a turnover from stage 3 to stage 4 at the age of 21 years. The calculated empiric distribution function showed 95% of stage 4 over 21 years while 75% of the patients with stage 3 were under 21 years. A reconstruction kernel suitable for osseous structures should be used, images should be viewed or presented in a bone window. CONCLUSION: According to these results it can be concluded that a person with stage 4 is probably 21 years or older, while a stage 3 leads to an estimated age under 21 years. On the other hand, a confidence level of 99.67% is not reached. Therefore, CT of the medial epiphysis of the clavicle will only be suitable for age estimation around the age of 21 years, if this relevant statistic obstacle is defeated. Bearing this in mind, further studies are needed to evaluate slice thickness as the most critical parameter. PMID- 16046093 TI - Taste disturbance after tonsillectomy and laryngomicrosurgery. AB - OBJECTIVE: To investigate the incidence of taste disturbance after tonsillectomy and laryngomicrosurgery. STUDY DESIGN: A prospective study. METHODS: Thirty-five consecutive patients underwent tonsillectomy with electric coagulator at Kanebo Memorial Hospital from September 2002 to May 2004. Another 35 consecutive patients underwent laryngomicrosurgery in a routine way during the same period. Both procedures were performed by two experienced doctors. The postoperative taste function was evaluated with questionnaire, electrogustometry (EGM), and serum levels of Zn, Cu and Fe. RESULTS: After tonsillectomy, 3/35 patients (8.6%) complained of taste disturbance, due to the pressure on the tongue in two, and Zn deficiency after lack of intake in one. All three patients recovered taste within 1.5 months. None of our patients had taste disturbance due to surgical insult of the lingual branch of the glossopharyngeal nerve. After laryngomicrosurgery, only one patient (2.9%) complained of taste disturbance with elevation of the EGM threshold that resulted from the pressure on the tongue. In addition, abnormal sensation in the tongue occurred in 9 cases (25.7%) after tonsillectomy and in 2 cases (5.7%) after laryngomicrosurgery, and feeling of thirst occurred in 12 cases (34.3%) and in 8 cases (22.9%), respectively. CONCLUSION: Taste disturbance occurs in few cases when experienced surgeons perform tonsillectomy or laryngomicrosurgery. However, it is recommended to inform the patients of the potential postoperative complications such as taste disturbance and abnormal sensation of the tongue. PMID- 16046094 TI - Stress views in the radiography of scapholunate instability. AB - AIM: On the suspected carpal instabilities stress views are recommended but not often used. The present study evaluates the reliability of the dorsal and volar stress radiographs on patients with posttraumatic wrist pain. PATIENTS AND METHODS: Stress radiographs of the wrists were examined in 22 patients with chronic wrist pain and the results were compared with scaphoid shift test and standard and positional views. The stress examination consists of applying to the wrist dorsal and volar stresses on the hand. RESULTS: Static scapholunate instability was diagnosed in 4 patients in whom 3 of them had positive scaphoid shift test sign as well. There were, however, 18 patients with dynamic scapholunate instability in whom the standard films were normal but dorsal stress radiography showed gap greater than 3mm between the scaphoid and lunate. CONCLUSION: Stress tests may provide considerable information in the evaluation of a patient who has a painful wrist in whom routine and special views do not demonstrate scapholunate dissociation. PMID- 16046095 TI - Purification of human tissue prokallikrein excreted from insect cells by liquid chromatography. AB - Tissue kallikrein, generally existing in living bodies as prokallikrein, is a serine proteinase that has proven of great significance to treat hypertension, cardiopathy and nephropathy. Although the extraction of tissue kallikrein from human urine is the most commonly used method to obtain such a protein, not only the yield is very little, but also the procedure is rather complex. Furthermore, the biological safety is uncertain. Therefore, the preparation of such a protein by genetic engineering method, including gene expression, cell culture, separation and purification, is very important. In this paper, a new method to obtain purified tissue prokallikrein excreted from insect cells by liquid chromatography has been proposed. In contrast to the previously published papers, the purification procedure is simplified to only three steps with the final yield of 57% and the purity of 95%, which is not only convenient, but also low-cost and suitable for the large-scale preparation of such a protein. The purified protein is further validated as prokallikrein by high performance liquid chromatography mass spectrometry and amino acid sequencing. PMID- 16046096 TI - Determination of ankle external fixation stiffness by expedited interactive finite element analysis. AB - Interactive finite element analysis holds the potential to quickly and accurately determine the mechanical stiffness of alternative external fixator frame configurations. Using as an example Ilizarov distraction of the ankle, a finite element model and graphical user interface were developed that provided rapid, construct-specific information on fixation rigidity. After input of specific construct variables, the finite element software determined the resulting tibial displacement for a given configuration in typically 15s. The formulation was employed to investigate constructs used to treat end-stage arthritis, both in a parametric series and for five specific clinical distraction cases. Parametric testing of 15 individual variables revealed that tibial half-pins were much more effective than transfixion wires in limiting axial tibial displacement. Factors most strongly contributing to stiffening the construct included placing the tibia closer to the fixator rings, and mounting the pins to the rings at the nearest circumferential location to the bone. Benchtop mechanical validation results differed inappreciably from the finite element computations. PMID- 16046097 TI - Effects of sarin on the operant behavior of guinea pigs. AB - The present study evaluated the dose-response effects of subacute exposure to sublethal doses of the organophosphorus (OP) chemical warfare nerve agent (CWNA) sarin (GB) on the operant behavior of guinea pigs. Dietary restricted guinea pigs, trained to respond for food under a progressive ratio (PR) schedule of reinforcement, were injected five times per week (Monday-Friday) for 2 weeks with fractions (0.1, 0.2, and 0.4) of the established LD(50) of GB (42 microg/kg). Changes in body weight, whole blood (WB) acetylcholinesterase (AChE) levels, and operant performances were monitored over the 2 weeks of GB exposure and for an additional 2 weeks following the termination of exposures. There were dose related changes in body weight and WB AChE levels throughout the exposure and post-exposure periods. Several parameters of PR performance were disrupted during exposure to 0.4 LD50 GB, however, concurrent weight loss indicated the presence of overt toxicity. PR performance recovered following the termination of exposures. Lower doses (0.1 and 0.2 LD50) of GB failed to produce reliable effects on operant performance during the exposure period. Overall responding decreased during exposure to 0.4 LD50 GB, resulting in reduced response rates and break points. The decrease in overall response rates was attributed to an increase in pausing since there was no decrease in running rate. Motor effects of 0.4 LD50 GB were evident as an increase in the proportion of lever press durations > or = 1.0 s. In the present study, doses of GB lower than 0.4 LD50 produced no marked alteration of operant performance in guinea pigs, although WB AChE levels were maximally inhibited to 20% of control. PMID- 16046098 TI - Global exponential stability of generalized recurrent neural networks with discrete and distributed delays. AB - This paper is concerned with analysis problem for the global exponential stability of a class of recurrent neural networks (RNNs) with mixed discrete and distributed delays. We first prove the existence and uniqueness of the equilibrium point under mild conditions, assuming neither differentiability nor strict monotonicity for the activation function. Then, by employing a new Lyapunov-Krasovskii functional, a linear matrix inequality (LMI) approach is developed to establish sufficient conditions for the RNNs to be globally exponentially stable. Therefore, the global exponential stability of the delayed RNNs can be easily checked by utilizing the numerically efficient Matlab LMI toolbox, and no tuning of parameters is required. A simulation example is exploited to show the usefulness of the derived LMI-based stability conditions. PMID- 16046099 TI - TNF-alpha G-308A polymorphism is associated with rheumatic fever and correlates with increased TNF-alpha production. AB - Previous studies suggested that abnormal regulation of TNF-alpha production may have a role in the pathogenesis of rheumatic fever (RF). Polymorphism at the promoter region of TNF-alpha gene (-308 A) has recently been shown to be associated with rheumatic heart disease (RHD) in Mexican patients. Although this polymorphism has long been shown to affect TNF-alpha gene expression in cell lines, its role in production of the cytokine in RF patients has not been studied. We therefore investigated TNF-alpha G-308A single nucleotide polymorphism and its effect on TNF-alpha production in 71 Turkish RF patients and 89 ethnically matched healthy controls. The TNF-alpha-308A allele frequency was found to be significantly higher in RF patients (RHD+arthritis) than in healthy controls [p<0.0032 Odds ratio (OR)=3.4, 95% confidence interval (CI) (1.5-7.7)]. When RHD patients were analyzed as a separate group, significant difference persisted [p<0.0055, OR=3.3, 95% CI (1.5-7.6)]. More importantly, ELISPOT analysis demonstrated that existence of A allele was associated with higher TNF alpha production compared with G allele. Our data suggest that carrying a high responder TNF-alpha-308A allele may be a genetic factor in increasing the susceptibility to develop RF disease. PMID- 16046100 TI - Ketogenic diet increases calbindin-D28k in the hippocampi of male ICR mice with kainic acid seizures. AB - The ketogenic diet (KD) increased the expression of calbindin-D(28k) (CB) in the interneurons of the hippocampus compared with the normal diet (ND)-fed mice. Also, 2 days after kainic acid (KA) administration, numerous CB-expressing astrocytes were found in the KD-fed mice compared with those of the ND-fed mice. These results suggest that the neuroprotective effect of the KD on the KA-induced toxicity may be, in part, mediated via an increased expression of CB. PMID- 16046101 TI - Comparison of gatifloxacin and levofloxacin administered at various dosing regimens to hospitalised patients with community-acquired pneumonia: pharmacodynamic target attainment study using North American surveillance data for Streptococcus pneumoniae. AB - This work aimed at determining the target attainment potential of gatifloxacin and levofloxacin in specific age-related patient populations such as elderly (> or =65 years) versus younger (<65 years) hospitalised patients with community acquired pneumonia (CAP). Previously described population pharmacokinetic models of gatifloxacin and levofloxacin administration in patients with serious CAP were utilised to simulate gatifloxacin and levofloxacin pharmacokinetics. Pharmacokinetic simulations and susceptibility data for Streptococcus pneumoniae from the ongoing national surveillance study, Canadian Respiratory Organism Susceptibility Study (CROSS), were then used to produce pharmacodynamic indices of free-drug area under the curve over 24h relative to the minimum inhibitory concentration (free-drug AUC(0-24)/MIC(all)). Monte Carlo simulations were then used to analyse target attainment both of gatifloxacin and levofloxacin to achieve free-drug AUC(0-24)/MIC(all)> or =30 against S. pneumoniae in patients with CAP. Dosing regimens for gatifloxacin were 400 mg once daily (qd) administered to younger patients (<65 years) and gatifloxacin 200 mg qd to elderly patients (> or =65 years). Dosing regimens for levofloxacin were simulated as 500 mg, 750 mg and 1000 mg qd administered to elderly patients as well as younger patients. Monte Carlo simulations using gatifloxacin 400mg against S. pneumoniae yielded probabilities of achieving free-drug AUC(0 24)/MIC(all) of 30 of 96.6% for all patients, 92.3% for younger patients and 97.7% for elderly patients. When administered to elderly patients, a reduced dose of gatifloxacin 200mg qd could achieve a target attainment potential of 91.4%. Monte Carlo simulation using levofloxacin 500 mg qd yielded probabilities of achieving free-drug AUC(0-24)/MIC(all) of 30 of 92.3% for all patients, 95.7% for elderly patients compared with 72.7% for younger patients. Using levofloxacin 750 mg and 1000 mg qd had probabilities of achieving free-drug AUC(0-24)/MIC(all) of 30 of 97.0% and 98.3%, 98.1% and 99.2%, and 90.1% and 95.2% for all patients, elderly patients and younger patients, respectively. The probability of achieving free-drug AUC(0-24)/MIC(all) of 100 was low both with gatifloxacin and levofloxacin, except in the case of elderly patients receiving levofloxacin in a dose of 1000 mg qd (78.5%). We conclude that gatifloxacin and levofloxacin pharmacokinetics in elderly patients with CAP are markedly different from those of younger patients. Higher gatifloxacin/levofloxacin AUC and longer half-life (t(1/2)) values in elderly patients with CAP compared with younger patients provide better pharmacodynamic parameters (free-drug AUC(0-24)/MIC) leading to a higher probability of pharmacodynamic target attainment and improved bacteriological outcome against S. pneumoniae. Gatifloxacin 400mg qd results in a high probability of target attainment and improved bacteriological outcome against S. pneumoniae both in young and elderly CAP patients. However, gatifloxacin administered at a lowered dose of 200 mg qd in elderly patients could still be successful in producing a favourable antibacterial effect. Levofloxacin administered at a dose of 750 mg qd results in a high probability of target attainment and improved bacteriological outcome against S. pneumoniae in all patients with CAP. PMID- 16046102 TI - Double-blind, placebo-controlled pharmacodynamic studies with a nutraceutical and a pharmaceutical dose of ademetionine (SAMe) in elderly subjects, utilizing EEG mapping and psychometry. AB - In a double-blind, placebo-controlled crossover study, the effects of S-adenosyl l-methionine (SAMe) on brain function measures of 12 normal elderly volunteers (6 m/6 f, aged 57-73 years, mean: 61 years) were investigated by means of EEG mapping and psychometry. In random order, the subjects were orally administered a pharmaceutical dose of 1600 mg SAMe, a nutraceutical dose of 400 mg SAMe and placebo, each over a period of 15 days, with wash-out periods of 2 weeks in between. EEG recordings, psychometric tests and evaluations of tolerability and side effects were carried out 0, 1, 3 and 6 h after drug administration on days 1 and 15. Multivariate analysis based on MANOVA/Hotelling T2 tests of quantitative EEG data demonstrated significant central effects of SAMe as compared with placebo after acute, subacute and superimposed drug administration of both the nutraceutical and the pharmaceutical dose. EEG changes induced by SAMe were characterized by an increase in total power, a decrease in absolute and relative power in the delta/theta and slow alpha frequencies, an increase in absolute and relative power in the alpha-2 and beta frequencies as well as an acceleration of the alpha centroid and the centroid of the total power spectrum. The delta/theta and the beta centroid showed variable changes over time. The dominant alpha frequency was accelerated, the absolute and relative power in the dominant alpha frequency attenuated after SAMe as compared with placebo. These acute and subacute pharmaco-EEG findings in elderly subjects are typical of activating antidepressants. Time-efficacy calculations showed that acute oral administration of SAMe in both the nutraceutical and the pharmaceutical dose induced the pharmacodynamic peak effect in the first hour with a subsequent decline. The 3rd and 6th hours still showed a significant encephalotropic effect after the 1600 mg dose. The maximum EEG effect was noted after 2 weeks of oral administration of both 1600 mg/die and 400 mg/die. The superimposed dose induced significant encephalotropic effects in the 3rd hour after 400 mg and in the 3rd and 6th hours after 1600 mg as compared with pre-treatment. Dose-efficacy calculations showed that the pharmaceutical dose of 1600 mg had a more pronounced effect on the CNS than the nutraceutical dose of 400 mg, with both doses being superior to placebo. Psychometric tests concerning noopsychic and thymopsychic measures as well as critical flicker fusion frequency generally demonstrated a lack of differences between SAMe and placebo, which reflects a good tolerability of the drug in elderly subjects. This was corroborated by the findings on side effects, pulse and blood pressure. PMID- 16046103 TI - The recognition potential: semantic processing or the detection of differences between stimuli? AB - The recognition potential is traditionally described as an electrical index elicited when subjects view a recognizable stimulus. Recent studies further show that it may be influenced by semantic processing. In this study, we investigated whether this observed influence is really produced by differences in semantic processing or whether it might be caused by the detection of differences between sequentially presented stimuli. In two different experiments, we systematically altered the type of background images presented while keeping the recognizable word constant. Analyses revealed that the same recognizable words elicited an RP with different amplitudes and latencies when viewed under different background conditions. Control stimuli, which were identical to background stimuli, did not elicit the RP. Hence, we postulate that when using the rapid stream stimulation paradigm, RP might also be influenced by the detection of differences between sequentially input stimuli. It is necessary to clarify whether RP changes are caused by the processing of the stimuli or by the detection of difference between successively input stimuli before any conclusion could be made. PMID- 16046104 TI - Immunoassay signal amplification on glass slides based on electroless deposition. AB - Saturated coverage of antibodies was first constructed on glass slides coated with polyelectrolyte multilayer; then sandwich immunoreactions were carried out. Following immunoreactions, the glass slides were immersed in a freshly prepared solution containing 0.01 wt% HAuCl4 and 0.4 mM NH2OH.HCl. The growth of immunogold nanoparticles on glass slides was monitored with UV-vis spectrometer in real time. The data indicated that the immunoassay signal amplification really depends on the time of electroless deposition process and the concentration of antigen. PMID- 16046105 TI - Adaptation and optimization of the emulsification-diffusion technique to prepare lipidic nanospheres. AB - In this study, the emulsification-diffusion method traditionally used to prepare polymeric nanoparticles was adapted to obtain lipidic nanospheres (LN) using four model lipids. The method consists of dissolving the lipid in a partially water miscible solvent (previously saturated with water) at room temperature or at controlled temperature depending on lipid solubility. This organic phase is emulsified in an aqueous solution of a stabilizing agent (saturated with solvent) by conventional stirring at the same temperature used to dissolve the lipid. This oil-in-water emulsion is then diluted with an excess of water at controlled temperature in order to provoke the diffusion from the internal phase into the external phase thereby causing lipid aggregation in the form of LN. This new approach for the preparation of LN has clear advantages over the existing methods, namely: (i) it is efficient and versatile; (ii) easy implementation and scaling up (with no need of high energy sources); (iii) high reproducibility and narrow size distribution; (iv) less physical stress (i.e., long exposure to high temperatures and to mechanical dispersion); (v) it is not necessary to dissolve the drug in the melted lipid. The selection of the water-miscible solvent and the stabilizers are critical parameters to obtain lipidic particles in the nanometric range. In general, solvents with high water miscibility and stabilizers able to form stable emulsions are preferred. The results demonstrated that it was possible to reduce the particle size by increasing the process temperature, the stirring rate, the amount of stabilizer, and by lowering the amount of lipid. Control of the preparative variables allowed to obtain LN with diameters under 100 nm. It was found that the influence of preparative parameters was associated with a mechanism based on a physicochemical instability. In this sense, it is suggested that the rapid solvent diffusion produces regions of local supersaturation near the interface, and LN are formed due to the ensuing interfacial phase transformations and lipid aggregation that occur in these interfacial domains. In terms of stability, only poly(vinyl alcohol) (PVAL) was able to preserve the physical stability of the dispersion for long periods after preparation. This effect was attributed to the ability of PVAL chains to form a strongly attached layer on the nanoparticle surface with an excellent repulsion effect. PMID- 16046106 TI - The rheological properties of self-emulsifying systems, water and microcrystalline cellulose. AB - The rheological properties of mixtures of equal parts of a range of ratios of a self-emulsifying system (MP) and water (W) added to microcrystalline cellulose (MCC), have been measured by an extrusion capillary rheometer. These measurements allow assessment of both the shear and tension components of flow plus the elastic behaviour of the wet powder masses, although the results for the estimation of shear stress require careful interpretation due to the limitation of the measuring system and the assumptions made in their derivation. The results indicate that there are three regions of behaviour of the systems, which are all significantly different from the mixtures containing only W and MCC. At low MP contents (1.5--23%), the masses increase in their resistance to shear and elongational flow and have lower elasticity. These similarities in behaviour occur in spite of considerable increase in the viscosity of the MPW mixtures and a change to non-Newtonian flow of the fluid. The behaviour of the 46% MP system is intermediate between these systems and the high MP concentrations (69, 80 and 92%). These latter systems show less resistance to shear and elongational flow than the first group of concentrations, but show considerably higher levels of elasticity. As the resistance to shear decreases, so does the impairment of the surface of the extrudate. There is clear evidence of a systematic change in behaviour of the wet powder masses as the values for the angle of entry of the wet mass into the die when plotted against the ratio of the resistance to die entry (upstream pressure loss) to the shear stress within the die, is linear on a log/log scale. Also, the values of compliance of the systems as a function of shear stress fall on a common curve. Changes in the ratio of the MPW to MCC for a system for a single level of MP (46%) resulted in a change in the values of the rheological parameters but not the type of behaviour. As all these wet powder masses had been shown previously to form pellets by the process of extrusion/spheronization, it is clear that systems with a wide range of rheological characteristics can be processed and no single rheological parameter can be used to provide complete characterisation of the processability of such systems. PMID- 16046108 TI - Direct or indirect? Graphical models for neural oscillators. AB - Univariate and bivariate time series analysis techniques have enabled new insights into neural processes. However, these techniques are not feasible to distinguish direct and indirect interrelations in multivariate systems. To this aim multivariate times series techniques are presented and investigated by means of simulated as well as physiological time series. Pitfalls and limitations of these techniques are discussed. PMID- 16046109 TI - [Intestinal parasitic infections in Togolese children under five years of age and PCIME recommendations]. PMID- 16046110 TI - [Ewing's tumor]. AB - Ewing's tumor (ET) is a malignant bone tumor occurring in children and young adults. ET affects mainly bones of the central axis, and almost always involves soft tissue infiltration. The discovery of a unique genetic alteration, which is a reciprocal translocation most frequently resulting in the fusion of the EWS gene situated on chromosome 22 with the FLI-1 gene on chromosome 11, currently places ET among neuroectodermal tumors. Moreover, this translocation is a tumor specific genetic marker at the basis of defining ET today and is used as a diagnostic and potentially prognostic tool complementary to imaging and histopathological work-up. Since the 1970 s, important progress has been made in the clinical management of ET patients. Multiagent chemotherapy in association with local treatment (surgery and/or radiation) has clearly improved outcome. The introduction of systemic treatment was justified by the frequent sub-clinical diffusion of apparently localized ET. Intensified therapeutic strategies have for the first time cured some metastatic ET patients, but at the cost of major side effects. Treatment is currently adapted as a result of a better definition of prognostic factors as well as a better assessment of its adverse effects. Improvement in global patient care and increased management of specific acute complications associated with ET (often interwoven with iatrogeneous effects) represent an important step towards improving the quality of life for ET patients as well as preventing long term complications. In the light of present studies, the majority of surviving adults today describe their health and quality of life as good. ET is a fascinating example of the progress made not only in the diagnostic and therapeutic approach to cancer but also in the comprehension of the mechanisms behind carcinogenesis, and consequently reflects the revolution of medicine over the last century. PMID- 16046111 TI - MKK3/6-p38 MAPK negatively regulates murine MMP-13 gene expression induced by IL 1beta and TNF-alpha in immortalized periodontal ligament fibroblasts. AB - Matrix metalloprotease-13 (MMP-13) or collagenase-3 is involved in a number of pathologic processes such as tumor metastasis and angiogenesis, osteoarthritis, rheumatoid arthritis and periodontal diseases. These conditions are associated with extensive degradation of both connective tissue and bone. This report examines gene regulation mechanisms and signal transduction pathways involved in Mmp-13 expression induced by proinflammatory cytokines in periodontal ligament (PDL) fibroblasts. Mmp-13 mRNA expression was increased 10.7 and 9.5 fold after stimulation with IL-1beta (5 ng/mL) and TNF-alpha (10 ng/mL), respectively. However, inhibition of p38 MAPKinase with SB203580 resulted in significant (p<0.001) induction (23.2 and 18.1 fold, respectively) of Mmp-13 mRNA as assessed by real time PCR. Negative regulation of IL-1beta induced Mmp-13 expression was confirmed by inhibiting p38 MAPK gene expression with siRNA. Transient transfection of dominant negative forms of MKK3 and MKK6 also resulted in increased levels of Mmp-13 mRNA after IL-1beta stimulation. Mmp-13 mRNA expression induced by TNF-alpha was decreased by JNK and ERK inhibition. Western blot and zymogram analysis indicated that Mmp-13 protein expression induced by the proinflammatory cytokines were also upregulated by inhibition of p38 MAPK. Reporter gene experiments using stable cell lines harboring 660-bp sequence of the murine Mmp-13 proximal promoter indicated that transcriptional mechanisms were at least partially involved in this negative regulation of Mmp-13 expression by p38 MAPK and upstream MKK3/6. These results suggest a negative transcriptional regulatory mechanism mediated by p38 MAPK and upstream MKK3/6 on Mmp-13 expression induced by proinflammatory cytokines in PDL fibroblasts. PMID- 16046112 TI - Lipoxins and lipoxin analogs in asthma. AB - The pathobiology of asthma is characterized by production of eicosanoids, a diverse family of bioactive fatty acids that play important roles in regulating airway inflammation and reactivity. Lipoxins (LXs) are products of arachidonic acid metabolism that are distinct from leukotrienes (LTs) and prostaglandins (PGs) in structure and function. Unlike the pro-inflammatory PGs and LTs, LXs display counter-regulatory actions. Cell-type specific biological actions have been uncovered for LXs and LX stable analogs that promote resolution of acute inflammatory responses. At least two classes of receptors, CysLT1 receptors and LXA4 receptors (named ALX), can interact with LXA4 and LXA4 analogs to mediate their biological actions. LXs are generated during asthma and LXA4 signaling blocks asthmatic responses in humans and experimental model systems. Of interest, respiratory diseases of increased severity, such as aspirin-intolerant asthma, cystic fibrosis and steroid-dependent, severe asthma, display defective generation of these protective lipid signals. Together, these findings indicate a pivotal role for LXs in mediating airway homeostasis. PMID- 16046113 TI - Epidemiological survey on incidence and treatment of community acquired pneumonia in Italy. AB - STUDY OBJECTIVES: To estimate annual incidence of community acquired pneumonia (CAP) in an Italian general population sample. DESIGN AND PARTICIPANTS: Two hundred and eighty-seven family practitioners (64.6% of those selected) recorded suspected or ascertained CAP cases for 1 year. Information on smoking habit, respiratory symptoms and signs, co-morbidity, antibiotic and corticosteroid therapy, hospitalization, mortality and recovery were obtained. RESULTS: Six hundred and ninety-nine case forms were collected (53.1% females, mean age 59.6+/ 19.5, 20.6% smokers). CAP incidence rates per 1000 population were: 1.69 in men vs. 1.71 in women; 2.33 in the North vs. 1.29 in the Centre-South of Italy; between 0.73 in 14-, and 3.34 in 64+year-old subjects. Main symptoms and signs were cough (73.3%), crackles (72.8%), dullness (57.3%), asthenia (53.4%). 59.5% of subjects had concurrent diseases, mostly cardiac and respiratory. 77.2% of cases had chest X-ray (with parenchymal density in 90.6%). Phlegm microbiological examination was performed in 12.8% of cases. First choice antibiotics were cephalosporins (45.8%), macrolides (20.2%), other beta-lactams (18.6%), and fluoroquinolones (12.2%). Rates of hospitalization and of mortality were 31.8% and 6.0%, respectively. CONCLUSION: This study confirmed that the annual CAP incidence rate in the general population of South Europe is about 2 per 1000 population and showed a wide choice of antibiotic treatment. PMID- 16046114 TI - Effect of carbon dioxide injection on production of wood cement composites from waste medium density fiberboard (MDF). AB - The possibility of recycling waste medium density fiberboard (MDF) into wood cement composites was evaluated. Both new fibers and recycled steam exploded MDF fibers had poor compatibility with cement if no treatment was applied, due to interference of the hydration process by the water soluble components of the fiber. However, this issue was resolved when a rapid hardening process with carbon dioxide injection was adopted. It appears that the rapid carbonation allowed the board to develop considerable strength before the adverse effects of the wood extractives could take effect. After 3-5 min of carbon dioxide injection, the composites reached 22-27% of total carbonation and developed 50 70% of their final (28-day) strength. Composites containing recycled MDF fibers had slightly lower splitting tensile strength and lower tensile toughness properties than those containing new fibers especially at a high fiber/cement ratio. Composites containing recycled MDF fibers also showed lower values of water absorption. Unlike composites cured conventionally, composites cured under CO(2) injection developed higher strength and toughness with increased fiber content. Incorporation of recycled MDF fibers into wood cement composites with CO(2) injection during the production stage presents a viable option for recycling of this difficult to manage waste material. PMID- 16046115 TI - Endometrial cancer: population attributable risks from reproductive, familial and socioeconomic factors. AB - In this study, based on the Swedish Family-Cancer Database, risk factors and their population attributable fractions (PAFs) for endometrial cancer were studied. Over 700,000 women at ages 51-68 years, accumulating 23 million person years at risk, were entered into Poisson analysis. Overall, reproductive factors (parity and age at last birth) showed a relative risk (RR) of 1.91 and a PAF of 45.51% when the reference group was women with a parity of 3+ and the last childbirth at ages over 34 years. The RR for family history was 2.33 but the PAF was only 2.09%. The RR for socioeconomic factors was a modest 1.12 but the PAF was 6.34%. The combined PAF of these three types of risk factors was 51.84%. Although the present analysis lacked data on some important risk factors for endometrial cancer, the results suggest that a large proportion of the etiology of endometrial cancer can be defined by known epidemiological risk factors. PMID- 16046116 TI - Influence of P-glycoprotein expression on in vitro cytotoxicity and in vivo antitumour activity of the novel pyrrolobenzodiazepine dimer SJG-136. AB - SJG-136 is a novel pyrrolobenzodiazepine dimer analogue that acts as a minor groove interstrand DNA cross-linking agent. The present study investigated the impact of ABCB1 (mdr-1) expression on the activity of SJG-136 using both in vitro and in vivo systems. SJG-136 was highly potent in the colon cancer cell lines HCT 116, HT-29 and SW620 (IC50 0.1-0.3 nM). However, HCT-8 and HCT-15 cells expressing significant levels of mdr-1 were less sensitive (IC50 2.3 and 3.7 nM, respectively) using a SRB assay. The cytotoxicity was increased in HCT-15 and A2780(AD) in presence of 5 microg/ml verapamil. Mdr-1 mRNA expression was determined by qRT-PCR and correlated to SJG-136 IC50s (r2=0.86, P=0.0001). Isogenic 3T3 cells expressing mdr-1 cDNA (3T3 pHamdr-1) were less sensitive to SJG-136 than the parental 3T3 cells (IC50 208 and 6.3 nM, respectively). Finally, SJG-136 (120 microg/kg/d dx5) was highly active against A2780 xenografts (SGD=275) but not A2780(AD) xenografts (SGD=67). PMID- 16046117 TI - Adjuvant endocrine therapy in postmenopausal women with early breast cancer: where are we now? AB - Tamoxifen has been the standard of care for adjuvant endocrine therapy of early breast cancer. In postmenopausal women, data now suggest that alternative agents (aromatase inhibitors [AIs]) may have improved long-term risk:benefit profiles and thus have the potential to improve outcome. The 'Arimidex', Tamoxifen, alone or in combination (ATAC) trial has shown that anastrozole provides improved disease-free survival (DFS) and time to recurrence, significantly reduced time to distant metastases and superior overall tolerability compared with tamoxifen when used as initial adjuvant therapy. Results have already led to a reconsideration of current recommendations for adjuvant therapy. Other ongoing trials include studies that are evaluating the benefits of sequencing of endocrine agents both within the standard 5-year adjuvant treatment period and as additional therapy in the post-adjuvant period. Three recently reported trials have suggested that switching from tamoxifen to an AI after 2-3 years of treatment leads to better outcomes than 5 years of tamoxifen. Finally, the NCIC MA 17 trial has shown that switching to an AI after 5 years of tamoxifen improves DFS compared with placebo. These are momentous discoveries that have improved our biological understanding and will inevitably change the management of breast cancer in the near future. PMID- 16046118 TI - Immunoregulation by 1,25-dihydroxyvitamin D3: basic concepts. AB - 1,25-Dihydroxyvitamin D(3) (1,25(OH)(2)D(3)), the biologically active metabolite of Vitamin D(3), not only regulates bone and calcium metabolism but also exerts other biological activities, including immunomodulation via the nuclear Vitamin D receptor expressed in antigen-presenting cells and activated T cells. This regulation is mediated through interference with nuclear transcription factors such as NF-AT and NF-kappaB or by direct interaction with Vitamin D responsive elements in the promoter regions of cytokine genes. Dendritic cells (DCs) are primary targets for the immunomodulatory activity of 1,25(OH)(2)D(3), as indicated by inhibited DC differentiation and maturation, leading to down regulated expression of MHC-II, costimulatory molecules and IL-12. Moreover, 1,25(OH)(2)D(3) enhances IL-10 production and promotes DC apoptosis. Together, these effects of 1,25(OH)(2)D(3) inhibit DC-dependent T cell activation. Immunomodulation by 1,25(OH)(2)D(3) and its analogs in vivo has been demonstrated in different models of autoimmune diseases and transplantation. Moreover, combining analogs with other immunosuppressants leads to synergism in models of autoimmunity and transplantation. The availability of 1,25(OH)(2)D(3) analogs with immunomodulatory activity at non-hypercalcemic doses may allow exploitation of their immunomodulatory effects in a clinical setting of treatment of autoimmune diseases and prevention of allograft rejection. PMID- 16046119 TI - Optimization of cultural and nutritional conditions for accumulation of poly-beta hydroxybutyrate in Synechocystis sp. PCC 6803. AB - Poly-beta-hydroxybutyrate (PHB) accumulation in the unicellular cyanobacterium, Synechocystis sp. PCC 6803, was studied under various cultural and nutritional conditions. Under controlled condition, cells harvested at the stationary phase of growth depicted maximum accumulation of PHB, i.e., 4.5% (w/w of dry cells) as compared to lag (1.8%) or logarithmic (2.9%) phases of cultures. A temperature range of 28-32 degrees C and pH between 7.5 and 8.5 were preferred for PHB accumulation. Cells cultivated under regular light-dark cycles accumulated more PHB (4.5%) than those grown under continuous illumination (2.4%). Nitrogen and phosphorus starvation stimulated PHB accumulation up to the tune of 9.5 and 11% (w/w of dry cells), respectively. Synechocystis cells pre-grown in glucose (0.1%) supplemented BG-11 medium when subjected to P-deficiency in presence of acetate (0.4%), PHB accumulation was boosted up to 29% (w/w of dry cells), the value almost 6-fold higher with respect to photoautotrophic condition. Fishpond discharges were found as suitable media for PHB accumulation in the test cyanobacterium. PMID- 16046120 TI - Diprolyl nitriles as potent dipeptidyl peptidase IV inhibitors. AB - Dipeptidyl peptidase IV (DPP4) is a multifunctional type II transmembrane serine peptidase which regulates various physiological processes, most notably plasma glucose homeostasis by cleaving peptide hormones glucagon-like peptide-1 and glucose-dependent insulinotropic polypeptide. Inhibition of DPP4 is a potentially valuable therapy for type 2 diabetes. Synthesis and structure-activity relationships of a series of substituted diprolyl nitriles are described, leading to the identification of compound 1 with a measured DPP4 K(i) of 3.6 nM. PMID- 16046121 TI - Novel peptide derivatives of bleomycin A5: synthesis, antitumor activity and interaction with DNA. AB - A series of novel amino acid and peptide derivatives of bleomycin (BLM) A(5) were synthesized. All the compounds possessed significant antitumor activities in vitro against HL-60, BGC-823, PC-3MIE8, and MDA-MB-435 cell lines. Their antitumor activities against MDA-MB-435 were 10-fold higher than BLM A5. The DNA cleavage studies indicated that the hydrophobic amino acid or peptide derivatives of BLM A5 could induce higher cleavage ratio of double to single strand DNA than BLM A5. From the DNA binding studies, we found that the derivatives containing either D-conformation amino acid or basic amino acid could facilitate DNA binding of BLM. PMID- 16046122 TI - Biphenyl-indanones: allosteric potentiators of the metabotropic glutamate subtype 2 receptor. AB - We have identified and synthesized a series of biphenyl-carboxylic acid indanones as allosteric potentiators of the metabotropic glutamate receptor 2. Structure activity relationship studies directed toward improving the potency and the brain to plasma ratio of the initial lead led to the discovery of 5 and 23 (EC50=111 and 5 nM, respectively). PMID- 16046123 TI - Discovery and structure-activity relationships of novel sulfonamides as potent PTP1B inhibitors. AB - A series of novel sulfonamides containing a single difluoromethylene-phosphonate group were discovered to be potent inhibitors of protein tyrosine phosphatase 1B. Structure-activity relationships around the scaffold were investigated, leading to the identification of compounds with IC50 or Ki values in the low nanomolar range. These sulfonamide-based inhibitors exhibit 100 and 30 times higher inhibitory activity than the corresponding tertiary amines and carboxamides, respectively. PMID- 16046124 TI - Modulation of base selectivity for a base-discriminating fluorescent nucleobase by addition of mercury ion. AB - We altered the fluorescence emission selectivity of a base-discriminating fluorescent base, (Py)U, from A-selective to T-selective by the addition of mercury ion. The strong fluorescence from a duplex containing the (Py)U/T base pair was specific to the mercury ion among divalent metal ions, providing a unique method for sensing mercury ions in aqueous solutions. PMID- 16046125 TI - Discovery of novel conformationally restricted diazocan peptidomimetics as inhibitors of interleukin-1beta synthesis. AB - A novel diazocan containing dipeptide mimetic was synthesized via reductive N-N bond cleavage of a pyrazolidino-pyrazolidine using Raney-Ni and evaluated as an ICE inhibitor. This versatile 8-membered ring containing scaffold possesses an N 5 ring nitrogen that was used to explore structure-activity relationships in a cell-based assay measuring inhibition of interleukin-1beta. PMID- 16046126 TI - Study on supramolecular complexing ability vis-a-vis estimation of pKa of substituted sulfonamides: dominating role of Balaban index (J). AB - The supramolecular complexing ability vis-a-vis pKa estimation of a large series of 43 sulfonamides was made using a series of molecular descriptors including topological indices. The set of topological indices chosen also contains Balaban (J) and a variety of Balaban type indices: J, Jz, Jm, Jv, Jc, and Jp. The results have shown that the most discriminating Balaban index (J) in multi-parametric regression analysis combined with indicator parameters yields excellent models and also establishes the superiority of the J index over other Balaban type indices. The statistics is improved when one of the indicator parameters is replaced by molar volume (MV). The results are discussed critically using a variety of statistics. PMID- 16046127 TI - Synthesis and in vitro study of novel 7-O-acyl derivatives of Oroxylin A as antibacterial agents. AB - A series of Oroxylin A derivatives, prepared by alkylation and condensation, were fully characterized by spectroscopic methods. All the derivatives were screened for antibacterial activity against a panel of susceptible and resistant Gram positive and Gram-negative organisms. It was observed that acylation of 7-OH group in Oroxylin A significantly enhanced the activity as compared to their parent compound (Oroxylin A). PMID- 16046128 TI - Squaric monoamide monoester as a new class of reactive immunization hapten for catalytic antibodies. AB - A squaric monoester monoamide motif was employed as an effective reactive immunogen for the discovery of monoclonal antibodies with reactive residue(s) in their combining sites. Two antibodies, 2D4 and 3C8, were uncovered that enhance paraoxon hydrolysis over background. Kinetic analysis of these antibodies was performed and interestingly both undergo a single turnover event due to covalent modification within the antibody combining site. Because antibodies 2D4 and 3C8 result in covalent attachment and thus inactivation of paraoxon, they could be useful probes for investigating paraoxon intoxication. PMID- 16046129 TI - Synthesis and evaluation of tricyclic pyrrolopyrimidinones as dipeptide mimetics: inhibition of interleukin-1beta-converting enzyme. AB - The application of a tricyclic pyrrolopyrimidinone scaffold for the synthesis of peptidomimetic inhibitors of interleukin-1beta-converting enzyme (ICE) is reported. The synthesis of the tricyclic scaffold and conversion of it to a variety of target ICE inhibitors were accomplished in 4-5 steps. In vitro biological evaluation of the tricyclic pyrrolopyrimidinones revealed fair to good ICE inhibitors, with the most active compound exhibiting an IC50 of 14 nM in a caspase-1 enzyme binding assay. PMID- 16046130 TI - Synthesis of aryl-1,2,4-triazine-3,5-diones as antagonists of the gonadotropin releasing hormone receptor. AB - Several efficient synthetic routes for 2-, 4-, and 6-aryl-1,2,4-triazine-3,5 diones were developed. Derivatives were synthesized and studied as gonadotropin releasing hormone antagonists in an effort to understand structure-activity relationships of the monocyclic compounds. PMID- 16046132 TI - Haemoglobin levels following unicompartmental knee arthroplasty: influence of transfusion practice and surgical approach. AB - A retrospective review was undertaken of preoperative and day 3 postoperative haemoglobin (Hb) levels in all unicompartmental knee arthroplasty (UKA) procedures performed by a single surgeon. Sixty-six UKAs were performed using the same prosthesis through an open approach with patella eversion. This group was compared with 212 UKAs performed using a minimally invasive approach without patella eversion, with an implant and instrumentation specifically devised for this approach. Both groups were well matched for patient demographics, surgical and anaesthetic techniques, thromboprophylaxis, and postoperative regimes. All patients received transfusions with pre-donated blood, except the unilateral minimally invasive approach group. An analysis of covariance was undertaken to examine the influence of the surgical approach and whether surgery was unilateral or bilateral, taking into account preoperative Hb levels and units of blood transfused. The average fall in Hb following UKA, adjusted for other variables, was 2.73 g/dl with an open approach compared to 1.82 g/dl with a minimally invasive approach. This difference was significant (p=0.0044). The average postoperative Hb in the minimally invasive group was 12.05 g/dl (range, 8.8 to 15.8 g/dl). Patients undergoing unilateral minimally invasive UKA are unlikely to develop symptomatic anaemia and should not be required to predonate blood or undergo transfusion. PMID- 16046133 TI - High tibial osteotomy: long term survival analysis and patients' perspective. AB - High tibial osteotomy has been generally accepted as a useful treatment for unicompartment osteoarthritis of the knee to improve a patient's quality of life. Few studies have examined the outcome from the patient's perspective. A survival analysis of one to twenty-one years was conducted on 67 knees including analysis of the patients' satisfaction level with this procedure. Cumulative survival probability of 89.5% at 5 years, 74.7% at 10 years and 66.9 % for 15 and 20 years was reported. Ninety-one percent of patients had improvement in pain score and would choose to have this surgery again. Average patient satisfaction level was 75.5%. Forty-eight percent of patients were able to perform at a higher level of activity than before surgery, although none was able to perform at the level prior to the onset of knee pathology. It was concluded that this procedure was able to improve a patient's quality of life and achieved high satisfaction among patients. There is an increasing role of high tibial osteotomy as an adjunct to an autologous chondrocyte implantation procedure. PMID- 16046134 TI - Preparation of transition-state analogues of sterol 24-methyl transferase as potential anti-parasitics. AB - There is an urgent need for new drugs to treat leishmaniasis and Chagas disease. One important drug target in these organisms is sterol biosynthesis. In these organisms the main endogenous sterols are ergosta- and stigmata-like compounds in contrast to the situation in mammals, which have cholesterol as the sole sterol. In this paper we discuss the design, synthesis and evaluation of potential transition state analogues of the enzyme Delta24(25)-methyltransferase (24-SMT). This enzyme is essential for the biosynthesis of ergosterol, but not required for the biosynthesis of cholesterol. A series of compounds were successfully synthesised in which mimics of the S-adenosyl methionine co-factor were attached to the sterol nucleus. Compounds were evaluated against recombinant Leishmania major 24-SMT and the parasites L. donovani and Trypanosoma cruzi in vitro, causative organisms of leishmaniasis and Chagas disease, respectively. Some of the compounds showed inhibition of the recombinant Leishmania major 24-SMT and induced growth inhibition of the parasites. Some compounds also showed anti parasitic activity against L. donovani and T. cruzi, but no inhibition of the enzyme. In addition, some of the compounds had anti-proliferative activity against the bloodstream forms of Trypanosoma brucei rhodesiense, which causes African trypanosomiasis. PMID- 16046135 TI - Cyclic peptide models of the Ca2+-binding loop of alpha-lactalbumin. AB - A series of cyclic peptides with different linkers were designed and synthesized to model the elbow-type Ca2+-binding loop of alpha-lactalbumin (LA). All amino acids of the Ca2+-binding loop are strikingly well conserved among LAs of different species with the sequence Lys79-Phe-Leu-Asp82-Asp-Asp-Leu-Thr- Asp87 Asp88, where three carboxylates of Asp82, Asp87, and Asp88 and the amide carbonyl oxygen atoms of Lys79 and Asp84 participate in Ca2+ binding. Alanine-containing models were also prepared for monitoring the role of the binding (82, 87-88) and nonbinding Asp residues (83-84) in coordinating the cation. The structural features of synthetic peptides and their Ca2+-binding properties were investigated in solution by circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy. In water, the CD curves show a strong negative band below 200 nm as a sign of the presence of unfolded conformers. In TFE, all cyclic peptides were found to have a CD spectrum, reflecting the presence of folded (turn) conformers. The effect of Ca2+ was dependent on the structure and concentration of the model and the Ca2+ to peptide ratio (r(cat)). A surprising time dependence of the FTIR spectra of Ca2+ complexes of the Ala-containing peptides was observed. The shape of the broad amide I band showed no more change after approximately 60 min. Contrary to this, the deprotonation of the side chain COOH group(s) and formation of the final coordination sphere of Ca2+ took more time. Infrared spectra showed that in the Ca2+ complex of model comprising the binding Asp residues of LA, the cation is coordinated to the COO- groups of all three Asps, while in the complex of model comprising nonbinding Asp residues of LA, the two neighboring Asp side chains form a bridged Ca2+-binding system. PMID- 16046136 TI - Chiral anthracene and anthrone templates as stereocontrolling elements in Diels Alder/retro Diels-Alder sequences. AB - The development of chiral anthracene templates for use in Diels-Alder/retro Diels Alder sequences is described. A summary of past results and new progress is reported. PMID- 16046137 TI - Ultrastructure of eggs of Paracapillaria (Crossicapillaria) philippinensis and evidence related to its life cycle. AB - The varied ultrastructure of the eggshell of Paracapillaria (Crossicapillaria) philippinensis, collected from a human sample, is reported from a scanning electron microscopy study. Two distinct egg shapes were identified: typical peanut-shaped and swollen peanut-shaped. Both thick and thin eggshells were detected. Thick eggshells are either fairly smooth or bear a beam-like network in relation to the pillars in their surface ultrastructure. Thin eggshells are transparent allowing visibility of the coiled larva within. Presence of the thin shell provides supportive evidence of autoinfection involved in the life cycle of this medically important parasite. PMID- 16046138 TI - Alternative separation methods of no-carrier-added 111In produced by heavy ion activation of silver. AB - 7Li3+ irradiation on natural silver target results in the formation of 111In. No carrier-added 111In was separated from bulk silver matrix by a column chromatographic method using the recently synthesized inorganic cation exchanger ceric vanadate as stationary phase and 10(-3)M HNO3 as mobile phase. PMID- 16046139 TI - Optically stimulated luminescence in forensics. AB - Tracking the former location of a radioactive source represents an attractive attribute in today's society, where the threat of a radiological dispersal device (RDD) is considered high. Development of a novel, retrospective detection technique based on optically stimulated luminescence is underway. This will allow confirmation of the former location of a radioactive source. Materials near the source's former location may be probed for information on accumulated dose by taking advantage of radiation-induced charge trapping that occurs in many common materials. The prototype detector design and analysis techniques are described. PMID- 16046140 TI - Coincident thresholds of mutant protein for paralytic disease and protein aggregation caused by restrictively expressed superoxide dismutase cDNA. AB - Familial amyotrophic lateral sclerosis (FALS) has been modeled in transgenic mice by introducing mutated versions of human genomic DNA encompassing the entire gene for Cu,Zn superoxide dismutase (SOD1). In this setting, the transgene is expressed throughout the body and results in mice that faithfully recapitulate many pathological and behavioral aspects of FALS. By contrast, transgenic mice made by introducing recombinant vectors, encoding cDNA genes, that target mutant SOD1 expression to motor neurons, only, or astrocytes, only, do not develop disease. Here, we report that mice transgenic for human SOD1 cDNA with the G37R mutation, driven by the mouse prion promoter, develop motor neuron disease. In this model, expression of the transgene is highest in CNS (both neurons and astrocytes) and muscle. The gene was not expressed in cells of the macrophage lineage. Although the highest expressing hemizygous transgenic mice fail to develop disease by 20 months of age, mice homozygous for the transgene show typical ALS-like phenotypes as early as 7 months of age. Spinal cords and brain stems from homozygous animals with motor neuron disease were found to contain aggregated species of mutant SOD1. The establishment of this SOD1-G37R cDNA transgenic model indicates that expression of mutant SOD1 proteins in the neuromuscular unit is sufficient to cause motor neuron disease. The expression levels required to induce disease coincide with the levels required to induce the formation of SOD1 aggregates. PMID- 16046141 TI - Apaf1 mediates apoptosis and mitochondrial damage induced by mutant human SOD1s typical of familial amyotrophic lateral sclerosis. AB - Several studies have indicated that apoptotic pathways are responsible for the loss of motor neurons that constitute the hallmark of amyotrophic lateral sclerosis (ALS). In this study, we demonstrate that apoptosis induced by the expression of several mutant Cu,Zn superoxide dismutases (SOD1) typical of familial ALS is mediated by Apaf1, a scaffold protein involved in neural development. Using different cell lines of neuronal origin and modulating the expression of both mutant SOD1s and Apaf1, we show that the removal of Apaf1 prevents cells death. Interestingly, intercepting activation of the caspases cascade is also effective in preventing both the mitochondrial damage and the increase in the production of reactive oxygen species induced by fALS-SOD1, even in the presence of cytochrome c release. This death pathway may be crucial also for the pathogenesis of the sporadic form of the disease, where markers of increased oxidative stress and mitochondria damage have been found. PMID- 16046142 TI - DNA methylating and demethylating treatments modify phenotype and cell wall differentiation state in sugarbeet cell lines. AB - In plants organogenesis, cell differentiation and dedifferentiation are fundamental processes allowing high developmental plasticity. Such plasticity involved epigenetic mechanisms but limited knowledge is available concerning quantitative aspects. Three sugarbeet (Beta vulgaris L. altissima) cell lines originating from the same mother plant and exhibiting graduate states of morphogenesis were used to assess whether these differences could be related or not to changes in DNA methylation levels. Methylcytosine percentages from 18.3 to 28.8% and distinct levels of DNA methyltransferase (EC 2.1.1.37) activities were shown in the three cell lines. The lowest methylcytosine percentage was associated to organogenesis. In order to test the plasticity of these cell lines, various treatments causing DNA hypo or hypermethylation were performed at different times and concentrations. In this collection of treated lines with+/ 10% of methylcytosine percentages, loss of organogenic properties and cell dedifferentiation were observed. As cell wall formation fits well with cell differentiation state, the lignification process was further investigated in treated and untreated lines as a biochemical marker of the phenotypic changes. For example, peroxidase specific activities (EC 1.11.1.7) varied from 0.7 to 0.02 pkat mg(-1) of protein in organogenic and dedifferentiated lines, respectively. A negative relationship between peroxidase activities, incorporation of cell wall bound phenolic compounds as ferulate and sinapate derivatives and methylcytosine percentages was obtained. This is the first biochemical evidence that phenotypic changes in plant cells induced by DNA hypo- or hypermethylating treatments are correlated in a linear relationship to modifications of the cell wall differentiation state. PMID- 16046143 TI - Probing the differential effects of infrared light sources IR1072 and IR880 on human lymphocytes: evidence of selective cytoprotection by IR1072. AB - Light therapy, both laser and LED, have been shown to provide clinical benefit in many therapeutic arenas. The effects of IR1072 and IR880 were investigated, using a range of single and multiple irradiation protocols, for their effect on freshly prepared human lymphocytes stimulated with phytohemagglutinin. Viable cell numbers remained significantly higher after irradiation with IR1072 and were significantly lower after IR880 irradiation compared to untreated controls, following a daily single irradiation over a 5-day period. Cell numbers were significantly higher after pre-treatment with IR1072 and exposure to UVA, compared to cells treated with UVA only. Cells irradiated twice on Day 3 post harvest with various wavebands confirm on Day 5, an increase in % cell viability after IR1072, and IR1072 alternating with IR1268 irradiation, and a decrease in % cell viability after IR880 irradiation alone. Further, wavebands tested displayed no significant differences compared to the control. Cells were collected after exposure on Days 3 and 5 with IR1072 and IR880 treatments and protein levels were compared using quantitative immunoblotting probed with an anti-iNOS antibody. Following IR1072, but not IR880, treatment there was a 4.9+/-2.1-fold higher iNOS protein expression in treated cells compared to the control on Day 5 post treatment. PMID- 16046144 TI - Post-transplant cutaneous T-cell lymphomas. AB - Post-transplant cutaneous lymphomas are rare. Cutaneous T-cell lymphomas account for 30% of these lymphomas. The clinical appearance of the skin lesions is identical to cutaneous lymphomas observed in non-immunosuppressed patients, with infiltrated plaques, nodular and ulcerated tumors, but with an increased frequency of erythroderma. Standard histology and immunohistochemistry are also consistent with the features of mycosis fungoides and CD30+ cutaneous lymphomas observed in the general population. However, the pronostic differs from the usually favourable outcome of cutaneous T-cell lymphomas, as 8 out of the 13 patients of our series died, in less than 1 year for 6 of them. This unfavourable course appears to be the same as that observed for systemic T-cell lymphoma in transplant recipients. In contrast to post-transplant B-cell lymphomas (systemic and primary cutaneous), the link to a virus has not been demonstrated. The prognosis is also less favourable for post-transplant cutaneous T-cell lymphomas than for post-transplant cutaneous B-cell lymphomas. PMID- 16046145 TI - Assembly of the N-cadherin complex during synapse formation involves uncoupling of p120-catenin and association with presenilin 1. AB - N-cadherin is an adhesion receptor that participates in both interaction between immature pre- and postsynaptic neurons and in the stabilization and function of matured neuron-neuron synapses. To better understand how the N-cadherin complex contributes to synapse formation, we examined its distribution and composition during synapse formation in the chick ciliary neurons. It was found that at early phases of synaptogenesis, N-cadherin is distributed in small clusters on the cell surface and primarily associates with p120-catenin and beta-catenin. In contrast, as synaptic contacts matured, larger N-cadherin clusters were found localized adjacent to the active zone and associated with PS1 and gamma-catenin, while p120 and beta-catenin were dispersed among other cell regions, including axons. As it is known that PS1 binds gamma-catenin and that uncoupled p120-catenin can alter the cytoskeleton via its effect on Rho GTPases, these changes in the molecular composition of the N-cadherin complex (represented by the uncoupling of p120 catenin and association with PS1) may correspond to distinct functional states of the complex involved in synaptic maturation. PMID- 16046146 TI - The integrated response of the human cerebro-cerebellar and limbic systems to acupuncture stimulation at ST 36 as evidenced by fMRI. AB - Clinical and experimental data indicate that most acupuncture clinical results are mediated by the central nervous system, but the specific effects of acupuncture on the human brain remain unclear. Even less is known about its effects on the cerebellum. This fMRI study demonstrated that manual acupuncture at ST 36 (Stomach 36, Zusanli), a main acupoint on the leg, modulated neural activity at multiple levels of the cerebro-cerebellar and limbic systems. The pattern of hemodynamic response depended on the psychophysical response to needle manipulation. Acupuncture stimulation typically elicited a composite of sensations termed deqi that is related to clinical efficacy according to traditional Chinese medicine. The limbic and paralimbic structures of cortical and subcortical regions in the telencephalon, diencephalon, brainstem and cerebellum demonstrated a concerted attenuation of signal intensity when the subjects experienced deqi. When deqi was mixed with sharp pain, the hemodynamic response was mixed, showing a predominance of signal increases instead. Tactile stimulation as control also elicited a predominance of signal increase in a subset of these regions. The study provides preliminary evidence for an integrated response of the human cerebro-cerebellar and limbic systems to acupuncture stimulation at ST 36 that correlates with the psychophysical response. PMID- 16046147 TI - Spatiotemporal characteristics of form analysis in the human visual cortex revealed by rapid event-related fMRI adaptation. AB - The integration of local elements to coherent forms is at the core of understanding visual perception. Accumulating evidence suggests that both early retinotopic and higher occipitotemporal areas contribute to the integration of local elements to global forms. However, the spatiotemporal characteristics of form analysis in the human visual cortex remain largely unknown. The aim of this study was to investigate form analysis at different spatial (global vs. local structure) and temporal (different stimulus presentation rates) scales across stages of visual analysis (from V1 to the lateral occipital complex-LOC) in the human brain. We used closed contours rendered by Gabor elements and manipulated either the global contour structure or the orientation of the local Gabor elements. Our rapid event-related fMRI adaptation studies suggest that contour integration and form processing in early visual areas is transient and limited within the local neighborhood of their cells' receptive field. In contrast, higher visual areas appear to process the perceived global form in a more sustained manner. Finally, we demonstrate that these spatiotemporal properties of form processing in the visual cortex are modulated by attention. Attention to the global form maintains sustained processing in occipitotemporal areas, whereas attention to local elements enhances their integration in early visual areas. These findings provide novel neuroimaging evidence for form analysis at different spatiotemporal scales across human visual areas and validate the use of rapid event-related fMRI adaptation for investigating processing across stages of visual analysis in the human brain. PMID- 16046148 TI - Attributions on the brain: neuro-imaging dispositional inferences, beyond theory of mind. AB - People need to predict what other people will do, and the other person's perceived disposition is the preferred mode of prediction. People less often use, for example, shared social norms to explain another person's behavior. Social psychology's last half-century of research on attribution theory offers precise, validated paradigms for testing how people think about other people's minds. Neuro-imaging data from one classic attribution paradigm shows the unique priority given to inferring chronic, idiosyncratic dispositions (unique attitudes, individual personality, idiosyncratic intent), compared to other kinds of mental contents. Specifically, sentences describing behavior that is low in consensus across actors, low in distinctiveness across entities, and high in consistency over time (compared with the other 7 low-high combinations) uniquely elicits (a) person attributions and (b) activation in the superior temporal sulcus. Ignoring consensus, both low-distinctiveness, high-consistency combinations (compared to 6 remaining combinations) also activate the MPFC, consistent with decades of behavioral data showing that general social cognition neglects consensus information. Thus, activated areas converge with prior neuro imaging data on theory of mind and social cognition, but more precisely isolate the exact nature of the inferences that activate these areas. PMID- 16046149 TI - Brain activation during execution and motor imagery of novel and skilled sequential hand movements. AB - This experiment used functional magnetic resonance imaging (fMRI) to compare functional neuroanatomy associated with executed and imagined hand movements in novel and skilled learning phases. We hypothesized that 1 week of intensive physical practice would strengthen the motor representation of a hand motor sequence and increase the similarity of functional neuroanatomy associated with executed and imagined hand movements. During fMRI scanning, a right-hand self paced button press sequence was executed and imagined before (NOVEL) and after (SKILLED) 1 week of intensive physical practice (n = 54; right-hand dominant). The mean execution rate was significantly faster in the SKILLED (3.8 Hz) than the NOVEL condition (2.5 Hz) (P < 0.001), but there was no difference in execution errors. Activation foci associated with execution and imagery was congruent in both the NOVEL and SKILLED conditions, though activation features were more similar in the SKILLED versus NOVEL phase. In the NOVEL phase, activations were more extensive during execution than imagery in primary and secondary cortical motor volumes and the cerebellum, while during imagery activations were greater in the striatum. In the SKILLED phase, activation features within these same volumes became increasingly similar for execution and imagery, though imagery more heavily activated premotor areas, inferior parietal lobe, and medial temporal lobe, while execution more heavily activated the precentral/postcentral gyri, striatum, and cerebellum. This experiment demonstrated congruent activation of the cortical and subcortical motor system during both novel and skilled learning phases, supporting the effectiveness of motor imagery-based mental practice techniques for both the acquisition of new skills and the rehearsal of skilled movements. PMID- 16046150 TI - Molecular phylogenetics of 'river dolphins' and the baiji mitochondrial genome. AB - It is well known that the classical river dolphins are not a natural group, but up to now the phylogenetic relationships among them are not very clear because different views have been referred from different studies. In the present study, we determined the complete nucleotide sequence of the mitochondrial (mt) genome of the baiji (Lipotes vexillifer), the most endangered cetacean species, and conducted phylogenetic analyses for the classical river dolphins based on data from cetacean mitochondrial genomes available. In our analyses, the classical river dolphins split into two separate lineages, Platanista and Lipotes+(Inia+Pontoporia), having no sister relationship with each other, and the Platanista lineage is always within the odontocete clade instead of having a closer affinity to Mysticeti. The position of the Platanista is more basal, suggesting separate divergence of this lineage well before the other one. The Lipotes has a sister relationship with Inia+pontoporia, and they together formed the sister group to the Delphinoidea. This result strongly supports paraphyly of the classical river dolphins, and the nonplatanistoid river dolphins do represent a monophyletic grouping, with the Lipotidae as the sister taxa to (Iniidae+Pontoporiidae), and is well congruent with the studies based on short interspersed repetitive elements (SINEs). PMID- 16046151 TI - Amitriptyline and nortriptyline serum determination by micellar liquid chromatography. AB - INTRODUCTION: Amitriptyline and nortriptyline are tricyclic antidepressants which act by enhancing the actions of norepinephrine and serotonin caused by blocking the re-uptake of various neurotransmitters at the neuronal membrane. A micellar liquid chromatographic procedure was developed to determine these drugs in serum samples for use in clinical monitoring. METHODS: The chromatographic determination of these highly hydrophobic substances was carried out using a 0.15 M SDS-6% (v/v) pentanol buffered at pH 7, in a C18 column, and electrochemical detection at 650 mV. The flow-rate was 1.5 mL/min. The analysis time was 14 min. RESULTS: The limits of detection (ng/mL) in serum were 0.25 and 0.31 for amitriptyline and nortriptyline, respectively. Repeatability and intermediate precision were evaluated at three different concentrations in serum samples. DISCUSSION: Untreated serum samples were injected directly into the HPLC system after filtration, leading to be a simple procedure that can be applied in routine analyses for Therapeutic Drug Monitoring. PMID- 16046152 TI - Impact of early hemispherotomy in a case of Ohtahara syndrome with left parieto occipital megalencephaly. AB - This report illustrates the usefulness and safety of very early hemispherotomy in an infant with Ohtahara syndrome (OS) secondary to left parieto-occipital megalencephaly. It provides evidence that surgical intervention might provide promising results in selected cases, and that young age is not a contraindication for this type of surgery. PMID- 16046153 TI - Low-frequency repetitive transcranial magnetic stimulation for seizure suppression in patients with extratemporal lobe epilepsy-a pilot study. AB - We evaluated the effect of low-frequency repetitive transcranial magnetic stimulation (rTMS) on seizure frequency in adult patients with medically intractable extratemporal lobe epilepsy (ETLE). Seven patients with medically intractable ETLE received low-frequency rTMS at 0.9 Hz, basically two sets of 15 min stimulation per day for five days in a week, with the stimulus intensity of 90% of resting motor threshold (RMT). The number of seizures during two weeks before and after the stimulation of one week was compared. Furthermore, RMT and active motor threshold (AMT) were measured before and after rTMS for each daily session. After low-frequency rTMS of one week, the frequency of all seizure types, complex partial seizures (CPSs) and simple partial seizures was reduced by 19.1, 35.9 and 7.4%, respectively. The patients with smaller difference between RMT and AMT before rTMS had higher reduction rate of CPSs. A favorable tendency of seizure reduction, though not statistically significant, during two weeks after low-frequency rTMS was demonstrated in medically intractable ETLE patients. As far as CPSs are concerned, smaller decrease of motor threshold by voluntary muscle contraction was associated with better response to rTMS. PMID- 16046154 TI - Anti-Mullerian hormone in premenopausal women and after spontaneous or surgically induced menopause. AB - OBJECTIVE: The objectives of this study were: (1) to determine anti-Mullerian hormone (AMH) levels in menopausal women, and (2) to confirm the source of AMH in cycling women and its disappearance after the removal of the source. METHODS: An observational and prospective study was conducted. RESULTS: A total of 47 women were recruited for the study. The study population consisted of the following groups of patients: (A) women of late reproductive age (n = 24; mean age +/- SD, 44 +/- 2.8 years); (B) menopausal women (n =14; mean age, 56 +/- 4 years); and (C) regularly cycling women undergoing surgical menopause (n = 9; mean age, 43 +/ 4 years). Blood samples were obtained from all patients. In patients undergoing surgery, blood samples were obtained before and after surgery. AMH was undetectable in 13 of 14 postmenopausal women, whereas it was undetectable in only two of 24 women of late reproductive age. A significant negative correlation has been found between AMH and age or follicle-stimulating hormone (FSH) in women of late reproductive age. In women who were candidates for oophorectomy, samples were obtained 3-5 days after surgery. AMH was undetectable after the surgery in all women. CONCLUSIONS: We found that AMH levels decreased in women in the late reproductive period and that menopause and ovariectomy in regularly cycling women are associated to undetectable AMH in serum. These observations confirm that the ovary could be the only source of AMH in women and that it is a novel marker for ovarian aging. PMID- 16046155 TI - First-trimester nuchal translucency screening in pregnant women who subsequently developed gestational diabetes. AB - OBJECTIVE: To compare the nuchal translucency (NT) thickness at 11-14 weeks of gestation in women who will later develop gestational diabetes mellitus (GDM) with that of women with normal glucose tolerance (NGT) in the second trimester of pregnancy. METHODS: Four hundred sixty-four women underwent NT screening at 11-14 weeks of gestation according to the standards of the Fetal Medicine Foundation. They all underwent an oral glucose tolerance test (OGTT) between 24 and 28 weeks. RESULTS: Women with GDM (n = 135) were significantly older (34.0 years [+/-4.8] vs 29.4 years [+/-5.2]; P < .001), had a significantly higher body mass index (BMI) (28.2 kg/m2 [+/-6.0] vs 24.4 [+/-5.0]; P < .001), and had a significantly higher serum concentration of hemoglobin A1c (HbA1c) at the time of the NT measurement (5.38% [+/-0.6] vs 4.95% [+/-0.4]; P < .001] than women with NGT (n = 329). There is no significant difference in the NT measurement at 11 to 14 weeks of gestation between women with GDM and NGT after correction for crown rump length (CRL) (r = -.08, P = .45). We performed multiple linear regression analysis with NT as the dependent variable, and BMI, maternal age, CRL, and GDM/NGT as independent variables. CRL (P < .001) was significantly related to the NT measurement, whereas GDM/NGT, BMI, and maternal age were not. CONCLUSION: As no significant difference in the NT measurement between women with GDM and NGT was observed, the risk estimation for chromosomal abnormality derived from the maternal age and NT measurement can also be used in women with glucose disorders. PMID- 16046156 TI - The effect of serum from women with preeclampsia on JAR (trophoblast-like) cell line. AB - OBJECTIVE: Pathologic placentation has been implicated in the pathogenesis of preeclamsia. We sought to assess the effect serum obtained from women with preeclampsia would have on JAR human choriocarcinoma cells regarding growth, invasiveness, and matrix metalloproteinase (MMP) secretion as compared to normotensive pregnant woman. METHODS: Blood was collected from 11 healthy pregnant women and from10 patients with preeclampsia at 28-33 weeks of gestation. The JAR human choriocarcinoma cell line was cultured in the presence of 10% serum obtained from each group. Cell proliferation, invasiveness, and MMP secretion was measured using a cell proliferation kit, the Matrigel (BD Biosciences, Beit Ha'Emek, Israel) invasion assay, and gel zymography, respectively. RESULTS: Cell growth increased by 6% when exposed to serum from patients with preeclampsia compared to 30% from controls (P <.01). Trophoblast invasion was significantly (P <.01) reduced in the preeclampsia group (21 +/- 1.9%) compared to controls (27 +/ 2.5%). Valid MMP-2 secretion was reduced by 51% in the preeclampsia group compared to controls (P <.05). CONCLUSION: Serum obtained from women with preeclampsia contains a factor or factors that exhibit an inhibitory effect on JAR trophoblast cell proliferation, invasiveness, and MMP-2 secretion. These factors may be involved in the pathologic placentation associated with the pathogenesis of preeclampsia. PMID- 16046157 TI - Complementary roles for the amygdala and hippocampus during different phases of appetitive information processing. AB - Evidence collected from rodent models of memory storage suggests that rapid forms of learning engage the involvement of multiple brain regions each of which may participate in a different component of information processing. The present study used temporary inactivation of the amygdala and hippocampus during different phases of information processing on a one-trial appetitive-conditioning task to examine how these two regions might participate in the storage of appetitive memories. Male Long Evans rats were chronically implanted into the amygdala or dorsal hippocampus and food deprived. Rats were trained on a radial maze conditioned cue preference task where training occurred in one 40-min session and testing took place 24 h later. The amygdala or hippocampus was inactivated separately with muscimol (50 ng/microl) injected immediately before or after training, or immediately before testing. Saline-injected rats displayed a conditioned preference by spending more time in the arm that previously contained food than in the arm that did not contain food. Muscimol injected into the amygdala before training or testing blocked the conditioned preference. Muscimol injected into the hippocampus immediately after training blocked the conditioned preference. These results suggest that the processing of memories may require multiple contributions from separate brain systems for at least short-term (24 h) storage. The resulting output from each system may converge on a similar downstream target to influence behavior. PMID- 16046158 TI - Structural model of the Plasmodium CDK, Pfmrk, a novel target for malaria therapeutics. AB - Malaria, with 300-500 million clinical cases resulting in 1-3 million fatalities a year, is one of the most deadly tropical diseases. As current antimalarial therapeutics become increasingly ineffective due to parasitic resistance, there exists an urgent need to develop and pursue new therapeutic strategies. Recent genome sequencing and molecular cloning projects have identified several enzymes from Plasmodium (P.) falciparum that may represent novel drug targets, including a family of proteins that are homologous to the mammalian cyclin-dependent kinases (CDKs). CDKs are essential for the control of the mammalian cell cycle and, based on the conservation of the CDKs across species, the plasmodial CDKs are expected to play a crucial role in parasitic growth. Here we present a 3D structural model of Pfmrk, a putative human CDK activating kinase (CAK) homolog in P. falciparum. Notable features of the present structural model include: (1) parameterization of the Mg2+ hexacoordination system using ab initio quantum chemical calculations to accurately represent the ATP-kinase interaction; and (2) comparison between the docking scores and measured binding affinities for a series of oxindole-based Pfmrk inhibitors of known activity. Detailed analysis of inhibitor-Pfmrk binding interactions enabled us to identify specific residues (viz. Met66, Met75, Met91, Met94 and Phe143) within the Pfmrk binding pocket that may play an important role in inhibitor binding affinity and selectivity. The availability of this Pfmrk structural model, together with insights gained from analysis of ligand-receptor interactions, should promote the rational design of potent and selective Pfmrk inhibitors as antimalarial therapeutics. PMID- 16046159 TI - Phosphodiesterase 4 regulation of cyclic AMP in pulmonary remodelling: potential roles for isoform selective inhibitors. PMID- 16046160 TI - Fasting and refeeding cause rapid changes in intestinal tissue mass and digestive enzyme capacities of Atlantic salmon (Salmo salar L.). AB - Fasting and refeeding effects on gastrointestinal morphology and digestive enzyme activities of Atlantic salmon, held in tanks of seawater at 9 degrees C and 31 per thousand salinity, were addressed in two trials. Trial 1: Fish (mean body mass 1190 g) were fasted for 40 days and intestines sampled at day 0, 2, 4, 11, 19 and 40. Trial 2: Fish (1334 g), fasted for 50 days, were refed and sampled at day 0, 3 and 7. Mass, length, protein, and maltase, lactase, and leucine aminopeptidase (LAP) activities were analyzed for stomach (ST), pyloric caeca (PC), proximal (PI), mid (MI), and distal intestine (DI). PC contributed 50% of gastrointestinal mass and 75% of enzyme capacity. Fasting decreased mass and enzyme capacities by 20-50% within two days, and 40-75% after 40 days. In PC, specific brush border membrane (BBM) maltase activity decreased whereas BBM LAP increased during fasting. Upon refeeding, enzyme capacities were mostly regenerated after one week. The results suggest that refeeding should start slowly with about 25% of estimated feed requirement during the first 3 days, but may then be stepped up rapidly. Investigations of digestive processes of fed fish should only be performed when intestines are feed-filled to avoid bias due to effects of fasting. PMID- 16046161 TI - Effects of dietary soybean protein levels on energy budget of the southern catfish, Silurus meridionalis. AB - Energy budgets were calculated for the southern catfish, Silurus meridionalis, fed diets replacing 0%, 13%, 26%, 39%, 52% and 65% fish meal protein with soybean meal (SBM) protein with or without methionine supplementation to apparent satiation at 27.5 degrees C. With increasing dietary soybean protein levels (SPL), the feed energy lost in feces, excretion and metabolism increased, while that available for growth decreased (P<0.05). When 0.12% or 0.26% methionine at 39% SPL was added to reach that in body carcass or the control group (0% SPL), no significant differences were found in each component of energy budgets. When 0.21% or 0.33% methionine at 52% SPL was added to reach the content of methionine in body carcass or the control group, energy spent on growth increased, but that on excretion and metabolism decreased (P<0.05). These results suggested that the differences in growth rate among the southern catfish fed the diets with different SPL were due to decreasing absorption rate, increasing excretion and metabolism with increasing dietary SPL. The most important factor limiting the use of soybean protein was the imbalance of essential amino acids, which resulted in more energy spent on metabolism and excretion, less energy on growth. Supplementation of methionine produced a relatively better amino acid profile and subsequently improved the utilization of soybean protein at high SPL, which resulted in less energy used for metabolism or lost in excretion and more energy available for growth. PMID- 16046162 TI - Carbohydrate metabolism of eggs of the whitefish, Coregonus spp. during embryogenesis and its relationship with egg quality. AB - The present study investigated the changes in carbohydrate metabolism of eggs of the whitefish, Coregonus spp. during embryogenesis (unfertilized eggs to embryos in the eyed stage). Occurrence of glycolysis was proved by activities of phosphofructokinase (PFK-1) and pyruvate kinase and by decreasing levels of hexose, pentose phosphate pathway by transaldolase (non-oxidative path) and glucose-6-phosphate dehydrogenase activities (oxidative path) and by increasing ribose levels, fructose synthesis (polyol pathway) by sorbitol dehydrogenase activities, gluconeogenesis by activities of glucose-6-phosphatase. Glycolysis and pentose phosphate pathway had highest activities up to the epiboly stage, gluconeogenesis from epiboly stage to the eyed embryo stage. Coregonus spp. eggs contained hexoses, ketoses, 6-deoxyhexoses, heptoses and uronic acids with hexoses, ketoses, and 6-deoxysugars occurring free and in bound form. Hexoses were found in highest quantities, followed by ketoses, and 6-deoxyhexoses. Levels of these compounds changed in a specific way during embryogenesis. During all investigated stages of embryogenesis, the levels of ribose, heptose, and ketose were correlated with the percentage of eyed stage embryos developing out of the fertilized eggs (egg viability). In distinct embryonic stages, the levels of hexoses and 6-deoxyhexoses and the activities of glucose-6-phosphatase were also correlated with egg quality. This ascertains the importance of carbohydrate metabolism for developing eggs. PMID- 16046163 TI - Maternal nutrient restriction during early to mid gestation up-regulates cardiac insulin-like growth factor (IGF) receptors associated with enlarged ventricular size in fetal sheep. AB - Intrauterine undernutrition is associated with a high incidence of cardiovascular diseases in adulthood. We previously showed that maternal nutrient restriction during early to mid gestation produces ventricular enlargement, although the mechanism is unknown. We examined myocardial expression of insulin-like growth factor I (IGF-1), IGF-2, IGF binding protein 3 (IGFBP-3), IGF-receptor 1 (IGF-1R) and IGF-2R in fetal sheep with maternal undernutrition. Multiparous ewes were fed with 50% (nutrient-restricted, NR) or 100% (control-fed, C) of NRC requirements from day 28 to 78 of gestation. Some of NR and C ewes were euthanized on day 78, and the rest were fed 100% NRC requirements from day 79 to 135 of gestation. At necropsy on day 78 or day 135 of gestation, gravid uteri were recovered. mRNA expression of IGF-1 and IGF-2 in ventricles were measured with RT-PCR, and protein expression of IGF-1R, IGF-2R, IGFBP-3 was quantitated with Western blot. Crown-rump length was reduced and left ventricle was enlarged in NR fetuses on day 78. At day 135 after re-alimentation, ventricular weights were similar between the two groups although ventricular wall thicknesses were greater in NR than C fetuses. No difference was found in IGF-1, IGF-2 or IGFBP-3 levels between the NR and C groups at either gestational age. Protein expression of IGF-1R and IGF-2R in the left ventricle and IGF-1R in the right ventricle was significantly elevated in the NR group on day 78 of gestation. Only IGF-1R expression remained elevated after late gestational re-alimentation in association with increases in ventricular wall thickness. Our study suggest that maternal undernutrition from early to mid gestation may change the expression of IGF-1R and IGF-2R in fetal myocardium, and play a role in cardiac ventricular enlargement in fetal sheep. PMID- 16046164 TI - Decoding Tat: the biology of HIV Tat posttranslational modifications. AB - The Tat protein is a viral transactivator that activates HIV transcription through complex interactions with RNA and host cell factors. Tat undergoes multiple posttranslational modifications that regulate the dynamics and complexity of these interactions. The biology of these modifications and their role in Tat function are reviewed. PMID- 16046165 TI - Peptides mimicking Vibrio cholerae O139 capsular polysaccharide elicit protective antibody response. AB - Vibrio cholerae is the etiological agent of cholera. V. cholerae serogroup O1 had been, until 1992, the only serogroup responsible for large epidemics and pandemics of cholera. In 1992, a new serotype of V. cholerae emerged in South East Asia that caused a massive outbreak of cholera in India and neighboring countries. The new serotype was named V. cholerae O139. The main differences between V. cholerae O139 and O1 are that the former possesses a capsular polysaccharide and different lipopolysaccharide. Capsular polysaccharides are, in general, T-independent antigens giving rise to poor immune responses lacking immunological memory. In order to overcome this, monoclonal antibodies against the capsular polysaccharide of V. cholerae O139 were used to screen different phage-displayed random peptide libraries. Eight different phage clones were selected and characterized using enzyme immunoassay with the monoclonal antibodies, and then tested for specificity by competition with V. cholerae O139 capsular polysaccharide. Selected peptides were sequenced, synthesized and conjugated to bovine serum albumin (BSA) and keyhole limpet hemocyanin (KLH). The conjugated peptides were used to immunize mice. It is evident that the anti peptide mouse antibodies bind to the V. cholerae O139 capsular polysaccharide. In addition, the anti-peptide antibodies are protective in a suckling mouse model. The protective efficacy is both specific and dose-dependent. A PCT (PCT/IT2003/000489) with the publication number WO 2004/056851 has been filed for the sequences of the eight peptides. PMID- 16046166 TI - Identification of macrophage induced genes of Corynebacterium pseudotuberculosis by differential fluorescence induction. AB - Corynebacterium pseudotuberculosis is the etiological agent of the sheep disease caseous lymphadenitis. We have developed a promoter reporter system for this organism based on expression of the green fluorescent protein (gfp) gene from Aequorea victoria. A promoterless vector, pSM20, containing the gfp gene was constructed, and promoters were inserted upstream of the gfp gene. Upon transformation into C. pseudotuberculosis, fluorescence could be visualised by fluorescence microscopy, and relative promoter strength measured by flow cytometry. The usefulness of this system for measuring changes in gene expression was demonstrated by measuring fluorescence levels of heat shocked C. pseudotuberculosis carrying a dnaK promoter construct. Replication of C. pseudotuberculosis within J774 macrophages could be monitored by fluorescence microscopy. The establishment of the system allowed the use of differential fluorescence to identify genes that showed up-regulation following macrophage infection. Genes coding for a non-ribosomal peptide synthetase and the beta chain of a propionyl CoA carboxylase were identified as possessing promoters that demonstrated enhanced activity following macrophage infection by C. pseudotuberculosis. PMID- 16046167 TI - Purification, potency and immunogenicity analysis of Vero cell culture-derived rabies vaccine: a comparative study of single-step column chromatography and zonal centrifuge purification. AB - Continuous Vero cell lines are more suitable for large-scale production of rabies vaccine. The purification of Vero cell-derived rabies vaccine is critical because of the residual cellular DNA and serum proteins. The perfection of techniques using column chromatography with different matrix material, gel filtration and zonal centrifugation is of paramount importance for the optimal purification of rabies vaccine, leaving minimal residual cellular DNA, below the permissible level of 100 pg per dose and serum protein content of 1 ppm. In this study the potency, immunogenicity and safety of Vero cell-derived rabies vaccines were compared following purification by densely or loosely packed DEAE-sepharose CL-6B columns with different bed heights or by zonal centrifugation. The optimal virus recovery and maximum removal of substrate DNA and serum proteins were achieved only when the sepharose CL-6B column bed height was maintained at a thickness of 2-2.5 cm. The rabies virus material was purified by layering over the matrix without applying pressure. DEAE-sepharose CL-6B column purification using a simplified, cost effective technique as described in this study enhances the antigen yield by 50% in comparison with zonal purification. PMID- 16046168 TI - Characterization of estrogen-responsive epithelial cell lines and their infectivity by genital Chlamydia trachomatis. AB - Chlamydial attachment and infectivity in vitro and ascending disease and sequelae in vivo have been reported to be enhanced/modulated by estrogen. Endometrial carcinoma cell lines Ishikawa and HEC-1B and the breast cancer lines MCF-7 and HCC-1806 were examined for Chlamydia trachomatis E infectivity. Estrogen receptor (ER) presence was confirmed by Western blot and qRT-PCR analyses. FACS analysis was used to determine the percent of plasma membrane-localized ERs (mERs), and their activity was tested by estrogen binding and competitive estrogen antagonists assays. Chlamydiae grew in all cell lines with HEC (90%) >> MCF-7 (57%)>Ishikawa (51%) >> HCC-1806 (20%). The cell line ER isoform composition was re-defined as: ERalpha + ERbeta + for MCF-7, HCC-1806 and Ishikawa; and ERbeta only for HEC-1B. HeLa cells were also tested and found to express ERbeta, but not ERalpha. A small percentage of both ERs were surface-exposed and functionally active. The endometrium-predominant ERbeta isoform was found in all cell lines, including those most representative of the common sites of C. trachomatis infection. Thus, the role of chlamydial attachment/infectivity will now be analyzed in ERbeta+and-isogenic HEC-1B cells. PMID- 16046169 TI - Perpetuation of immunological memory: role of serum antibodies and accessory cells. AB - A self-sustaining mechanism for perpetuation of immunological memory is proposed which requires the involvement of idiotypic-anti-idiotypic antibody immune complexes, antigen-presenting cells and follicular dendritic cells, but not long lived memory cells or persisting antigens. PMID- 16046170 TI - Visceral leishmaniasis in organ transplant recipients: 11 new cases and a review of the literature. AB - Eleven new cases of visceral leishmaniasis (VL) are reported in organ transplant patients in France. The epidemiological, clinical, biological, diagnostic and therapeutic features are reviewed, based on these cases and 46 cases reported in the literature. VL was most commonly associated with renal transplantation (77% of the cases). Most patients were from Southern European countries. The main clinical symptom was fever. Leucopoenia and anaemia were the most frequent haematological disorders. Diagnosis was by direct finding of the parasite in smears of bone marrow (85.2%) or, by positive serology (90.9%). Without antileishmanial treatment, VL in transplant recipients was fatal. Treatment using either antimonials or amphotericine B gave similar cure rates of around 80% of the cases. But toxicity was higher for antimonials. Relapses occurred in 14.3%. PMID- 16046171 TI - Pathophysiology of disk-related low back pain and sciatica. II. Evidence supporting treatment with TNF-alpha antagonists. AB - Strong evidence suggests that TNF-alpha may be among the chemical factors involved in disk-related sciatica. TNF-alpha is involved in the genesis of nerve pain in animal models and may promote pain-signal production from nerve roots previously subjected to mechanical deformation. In animal experiments, TNF-alpha has been identified in nucleus pulposus and Schwann cells. Local production of endogenous TNF-alpha may occur early in the pathogenic process. Exposure to exogenous TNF-alpha induces electrophysiological, histological, and behavioral changes similar to those seen after exposure to nucleus pulposus, and these changes are more severe when mechanical compression is applied concomitantly. TNF alpha antagonists diminish or abolish abnormalities in animal models. Other cytokines may be involved also, as suggested by the potent inhibitory effects of compounds such as doxycycline. Two open-label studies in humans suggest dramatic efficacy of TNF-alpha antagonists in alleviating disk-related sciatica. In contrast, the results of the only controlled study available to date do not support a therapeutic effect of TNF-alpha antagonists. Thus, whether TNF-alpha antagonist therapy is warranted in patients with disk-related sciatica remains an open question, and further randomized controlled studies are needed. PMID- 16046172 TI - Septic knee arthritis after intra-articular hyaluronate injection. Two case reports. AB - Intraarticular sodium hyaluronate injection to treat osteoarthritis is associated with minor side effects. Infections seem uncommon. We report two cases of septic knee arthritis. One patient was an 80-year-old woman who was admitted for Staphylococcus aureus knee arthritis after several intraarticular injections of sodium hyaluronate and corticosteroids. In the other patient, a 78-year-old woman, Neisseria mucosa knee arthritis occurred after a single sodium hyaluronate injection. Faultless aseptic technique is essential when administering hyaluronate viscosupplementation. Patients should be informed of the risk of septic arthritis. PMID- 16046173 TI - Pathophysiology of disk-related sciatica. I.--Evidence supporting a chemical component. AB - Sciatica in patients with disk disease was long ascribed to pressure put on the sciatic nerve root by a herniated disk. However, a role for chemical factors acting in conjunction with this mechanical insult is suggested by a number of clinical observations: disk surgery does not consistently provide pain relief, large disk herniations are not always symptomatic, severe pain may be present in patients without imaging evidence of nerve root compression, the severity of symptoms and neurological signs is not well correlated with the size of the disk herniation, and conservative therapy is often effective. Experimental studies have provided further evidence for a chemical component: disk herniations can undergo spontaneous resorption, the intervertebral disk is immunogenic, and mediators for inflammation have been identified within intervertebral disk tissue. The current pathophysiological theory incriminates proinflammatory substances secreted by the nucleus pulposus (NP). When preexisting or concomitant mechanical injury to a nerve root occurs, these substances can cause nerve root pain. Animal experiments have established that the NP can induce functional and structural nerve root abnormalities in the absence of mechanical compression and that this effect is mediated by substances located at the surface of NP cells. Methylprednisolone, diclofenac, indomethacin, doxycycline, and cyclosporine induce variable inhibition of this effect. Available information points to tumor necrosis factor-alpha (TNF-alpha) as the main candidate among substances potentially responsible for nerve root pain. Therefore, trials of TNF-alpha antagonists in patients with disk-related sciatica are warranted. PMID- 16046174 TI - Bone mineral density in premenopausal women with major depression. AB - AIM: To investigate the relationship between the major depression and bone mineral density (BMD) in premenopausal women. MATERIAL AND METHODS: We compared BMD, plasma cortisol level, osteocalcin and C-telopeptide levels of 35 premenopausal women with major depression with those of 30 healthy women who were matched for age and body mass index. Major depression was diagnosed according to Diagnostic and Statistical Manual of Mental Disorders (fourth edition) criteria. Nineteen patients had mild and 16 patients had moderate severity of major depression as measured by Hamilton rating scale for depression. RESULTS: Women with any risk factor for osteoporosis were excluded from the study. All women underwent BMD measurement by DEXA at lumbar (L2-4) and femoral neck region. After an overnight fasting, plasma cortisol levels were measured at 08:00 h by using competitive immunoassay method. Osteocalcin and C-telopeptide were used for the evaluation of bone turnover. There were no significant differences in BMD, plasma cortisol level, osteocalcin and C-telopeptide levels between the patients and the control groups. There was also no correlation between the plasma cortisol level, the duration and the severity of disease, antidepressant drug use and BMD. CONCLUSION: Major depression had no significant effect on BMD and bone turnover markers in our patient group of mild to moderate severity of the disorder. PMID- 16046175 TI - Comparison of bone loss induced in female rats by hindlimb unloading, ovariectomy, or both. AB - INTRODUCTION: Clinical and experimental evidence supports a link between the effects of mechanical loading and those of estrogens on bone. The objective of this study was to compare bone loss induced in female rats by hindlimb unloading, ovariectomy, or both. MATERIALS AND METHODS: Thirty-six female Wistar rats aged 12 weeks were randomized to bilateral surgical ovariectomy without tail suspension (OV) or with tail suspension for 30 days (OV-TS) or to sham surgery without tail suspension (control group, C) or with tail suspension for 30 days (TS). Bone mineral density (BMD) of the distal femoral metaphysis was measured in g/cm2 by dual X-ray absorptiometry in all 12 animals on days 0, 7, 14, and 30. RESULTS: On D14 and D30, BMD (mean+/-S.D.) was significantly lower in the OV, TS, and OV-TS groups than in the control group (D14: 0.239+/-0.014, 0.243+/-0.016, and 0.227+/-0.018, respectively, vs. 0.258+/-0.005 in the controls; P<0.05; and D30: 0.241+/-0.011, 0.227+/-0.015, and 0.200+/-0.018, respectively, vs. 0.279+/ 0.009 in the controls; P<0.001). On D30, the percentage BMD change versus baseline (mean+/-S.D.) differed significantly between the combination (OV-TS) group (-14.26+/-8.14) and the single-intervention groups (OV: +0.99+/-6.44, P<0.001; and TS: -6.36+/-4.56, P<0.05). As early as D7, bone loss was significantly greater in the combination (OV + TS) group than in the OV group ( 1.79%+/-7.17 vs. +4.29%+/-9.55; P<0.05). CONCLUSION: In female rats, the rate and severity of bone loss were greater when estrogen deprivation was combined with mechanical unloading than when either intervention was used alone. Mechanical unloading induced a greater degree of bone loss than did estrogen deprivation. In this model of high-rate bone loss, mechanical unloading may predominate over estrogen deprivation in the genesis of bone loss. PMID- 16046176 TI - Biphasic surface amorphous layer lubrication of articular cartilage. AB - The biphasic nature of articular cartilage has been acknowledged for some time and is known to play an important role in many of the biomechanical functions performed by this unique tissue. From the lubrication point of view however, a simple biphasic model is unable to account for the extremely low friction coefficients that have been recorded experimentally, particularly during start up. In addition, research over the last decade has indicated the presence of a surface amorphous layer on top of articular cartilage. Here, we present results from a finite element model of articular cartilage that includes a thin, soft, biphasic surface amorphous layer (BSAL). The results of this study show that a thin BSAL, with lower elastic modulus, dramatically altered the load sharing between the solid and liquid phases of articular cartilage, particularly in the near-surface regions of the underlying bulk cartilage and within the surface amorphous layer itself where the fluid load support exceeded 85%. By transferring the load from the solid phase to the fluid phase, the biphasic surface layer improves lubrication and reduces friction, whilst also protecting the underlying cartilage surface by 'shielding' the solid phase from elevated stresses. The increase in lubrication effectiveness is shown to be greatest during short duration loading scenarios, such as shock loads. PMID- 16046177 TI - Demystifying biomedical signals: a student centred approach to learning signal processing. AB - The processing and analysis of physiological signals has become firmly established in clinical medicine and biomedical research. Many of the users of this technology however do not come from an engineering or science background, and traditional approaches in teaching signal processing are thus not appropriate for them. We have therefore developed a series of modular courses that are aimed specifically at an audience with a background in medicine, health-care or the life-sciences. In these courses, we focus on the concepts, principles and rationale of applying signal processing methods, rather than the mathematical foundations of the techniques. Thus, we aim to remove some of the perceived 'mystery' often surrounding this subject. The very practical approach, with hands on experience using the MATLAB software, has been well received, with strong evidence that students have learnt to apply their knowledge. This paper describes the learning and teaching approach taken, and some of the experience acquired. PMID- 16046178 TI - e-Learning system ERM for medical radiation physics education. AB - The objective of this paper is to present the Education for Radiation in Medicine (ERM) e-Learning System. The system was developed, tested and piloted in the Inter-University Medical Physics Centre, Plovdiv, Bulgaria. It was based on the results of EU Project TEMPUS S-JEP 09826. The ERM e-Learning System is an integrated on-line system for remote education covering aspects of Medical Radiation Physics education (M.Sc. level). It provides user-friendly interface and optimised functionality with three different access levels: trainee, professor and administrator. The minimum server requirements and the standard client side working environment turn the system into a good, cost effective and easy to support solution for remote education. PMID- 16046179 TI - KISS--a new approach to self-controlled e-learning of selected chapters in Medical Engineering and other fields at bachelor and master course level. AB - Modern life style requires new methods for individual lifelong learning, based on access at every time and from every place. This fundamental requirement is provided by the Internet. The Internet technology promises an increasing potential in the future for e-learning or tele-learning. Some special requirements are password-controlled access, applicability of most commercially available PCs and laptops equipped with standard software (Microsoft Internet Explorer 6.0), central evaluation of the students' performance, inclusion of an examination part, provision of a picture gallery and a comprehensive glossary accessible in the learning mode. The KISS-shell has been developed based on the Oracle 10g application server in combination with a relational data base (Oracle 8i) on the server side and a web browser based interface using JavaScript for user control of data input on the client side (Kontrolliertes Intelligentes Selbstgesteuertes Studium, KISS). The first tutorial application has been realized with a chapter about cardiac pacemakers. The weight of that chapter (or module) is about 2 ECTS (i.e. the equivalent of 30 working hours; European Credit Transfer System, ECTS). The internal structure of the chapter is organized in sequential mode. It consists of five main sections. Each of those five sections is subdivided into five subsections of comparable length. Progression from one subsection to the next is possible only after successfully passing through the respective examination. The whole learning programme with the pacemaker chapter has been evaluated by 10 students. The system will be presented together with first experiences including the evaluation results. Until now the program has not been used for training purposes. PMID- 16046180 TI - CXC chemokines in angiogenesis. AB - CXC chemokines display pleiotropic effects in immunity, regulating angiogenesis, and mediating organ-specific metastases of cancer. In the context of angiogenesis, CXC chemokines are a unique family of cytokines, known for their ability to behave in a disparate manner in the regulation of angiogenesis. Members that contain the 'ELR' motif are potent promoters of angiogenesis, and mediate their angiogenic activity via binding and activating CXCR2 on endothelium. In contrast, members, in general, those are inducible by interferons and lack the ELR motif (ELR-) are potent inhibitors of angiogenesis, and bind to CXCR3 on endothelium. This review will discuss the biology of these angiogenic and angiostatic CXC chemokines and discuss their disparate angiogenic activity in the context of a variety of disorders. PMID- 16046181 TI - An EM algorithm for shape classification based on level sets. AB - In this paper, we propose an expectation-maximization (EM) approach to separate a shape database into different shape classes, while simultaneously estimating the shape contours that best exemplify each of the different shape classes. We begin our formulation by employing the level set function as the shape descriptor. Next, for each shape class we assume that there exists an unknown underlying level set function whose zero level set describes the contour that best represents the shapes within that shape class. The level set function for each example shape in the database is modeled as a noisy measurement of the appropriate shape class's unknown underlying level set function. Based on this measurement model and the judicious introduction of the class labels as the hidden data, our EM formulation calculates the labels for shape classification and estimates the shape contours that best typify the different shape classes. This resulting iterative algorithm is computationally efficient, simple, and accurate. We demonstrate the utility and performance of this algorithm by applying it to two medical applications. PMID- 16046182 TI - Candida yeast long chain fatty alcohol oxidase is a c-type haemoprotein and plays an important role in long chain fatty acid metabolism. AB - The industrial yeasts Candida tropicalis or Candida cloacae are able to grow on a variety of long chain alkanes and fatty acids as the sole carbon source. The complete oxidation of these substrates involves two sequential oxidative pathways: omega-oxidation, comprising the P450 alkane oxidase, a flavin-dependent membrane-bound long chain fatty alcohol oxidase [FAO] and a possible separate aldehyde oxidase [F.M. Dickinson, C. Wadforth, Purification and some properties of alcohol oxidase from alkane-grown Candida tropicalis, Biochem. J. 282 (1992) 325-331], and the beta-oxidation pathway, which utilises acylCoA substrates. We recently purified the membrane-bound long chain fatty alcohol oxidase FAO1 and confirmed it is also a c-type haemoprotein. Multiple isoforms may exist for many of these long chain fatty alcohol oxidases and the in vivo requirements for individual genes with respect to specific substrates are still being elucidated. In vitro reconstitution experiments have demonstrated that in Candida maltosa, the cytochrome P450 52A3 gene product can completely oxidise alkanes to dicarboxylic acids [U. Scheller, T. Zimmer, D. Becher, F. Schauer, W. Schunck, Oxygenation Cascade in Conversion of n-Alkanes to, -Dioic Acids Catalyzed by Cytochrome P450 52A3, J. Biol. Chem. 273 (1998) 32528-32534], potentially obviating requirements for a long chain alcohol oxidase. Here, we directly determine in vivo the role of the long chain alcohol oxidase (FAOT) in C. tropicalis, grown on a variety of substrates, followed by gene deletion. The faot double knockout has no detectable faot activity and is incapable of growth on octadecane, but it grows well on oleic acid, palmitic acid and shorter chain alkanes/fatty acids. A spontaneous mutation[s] may have occurred in the faot double gene knockout of C. tropicalis resulting in its inability to grow on oleic acid and hexadecane. The mutations demonstrate that different pathways of octadecane, hexadecane, oleic acid and palmitic acid utilisation exist in C. tropicalis. PMID- 16046183 TI - Significance of bed porosity, bran and specific surface area in solid-state cultivation of Aspergillus oryzae. AB - In this paper, the effects of bed porosity, bran and specific surface area on the oxygen uptake rate and alpha-amylase production during growth of Aspergillus oryzae on wheat grain and wheat-flour substrate are reported. The high oxygen uptake rate found during cultivation of A. oryzae on wheat-flour substrate was not reached on wheat grain. This is mainly due to the bran of the wheat grain. Using wheat-flour substrates, it was shown that extra bed porosity increased the alpha-amylase production and oxygen uptake rates. Furthermore, the peak oxygen uptake rate decreased with increasing surface area-volume ratio of the substrate particles, while the alpha-amylase production and the cumulative oxygen uptake per gram of initial substrate dry matter increased. The present work does not support a direct correlation between aerial mycelia and enzyme production. There is, however, a correlation between the alpha-amylase yield and the cumulative oxygen uptake (not the uptake rate). This implies that aerial mycelia could accelerate alpha-amylase production even if they do not increase the yield. PMID- 16046184 TI - Unravelling Cryptosporidium and Giardia epidemiology. AB - Molecular biology has provided insights into the taxonomy and epidemiology of Cryptosporidium and Giardia, which are major causes of protozoal diarrhoea in humans worldwide. For both genera, previously unrecognized differences in disease, symptomatology, zoonotic potential, risk factors and environmental contamination have been identified using molecular tools that are appropriate for species, genotype and subtype analysis. In this article, to improve understanding of the epidemiology of cryptosporidiosis and giardiasis, we consider specific requirements for the development of more-effective molecular identification and genotyping systems that should be applicable to both clinical and environmental samples. PMID- 16046185 TI - Targeting malaria parasite proteins to the erythrocyte. AB - The intraerythrocytic stages of the protozoan parasite Plasmodium falciparum reside within a parasitophorous vacuole (PV) and set up unique "extraparasite, intraerythrocyte" protein-trafficking pathways that target parasite-encoded proteins to the erythrocyte cytoplasm and cell surface. Two recent articles report the identification of trafficking motifs that regulate the transport of parasite-encoded proteins across the PV. These articles greatly aid the annotation of the parasite "secretome" catalog of proteins that are targeted to the erythrocyte cytoplasm or cell membrane. PMID- 16046186 TI - Pexel/VTS: a protein-export motif in erythrocytes infected with malaria parasites. AB - The trafficking of proteins from the malaria parasite into the erythrocyte host has an important role in both the adaptation by the parasite of its immediate environment and the pathophysiology of disease. The molecular basis of these trafficking processes, particularly export from the parasitophorous vacuole (PV), is poorly understood and a matter of some controversy. In this article, we highlight two recent, independent reports that have provided new insights into protein translocation across the PV membrane, characterizing a novel signalling motif as a key signature in cataloguing the parasite secretome. PMID- 16046187 TI - Artemisinin resistance: how can we find it? PMID- 16046188 TI - Mitral annular motion as a surrogate for left ventricular function: correlation with brain natriuretic peptide levels. AB - BACKGROUND: Pulsed-wave (PW) Doppler tissue velocities of the mitral annulus correlate well with Left Ventricular (LV) diastolic(D) and systolic(S) functions. Brain natriuretic peptide (BNP) levels have been shown to be elevated in patients with symptomatic LV dysfunction (Dys) and correlate to the severity of symptoms and prognosis. OBJECTIVES: To validate the accuracy of mitral annular motion (MAM) assessed by Doppler Tissue Imaging (DTI) & M-mode Echocardiography (MME) as a surrogate for determination of LV function in comparison with BNP. METHODS: A series of 133 patients with a variety of cardiac pathologies referred for echocardiography and 20 healthy age & sex matched volunteers as a control group were included the study. Ejection fraction (EF) of LV, Doppler recordings of the mitral inflow, MME and PWDTI data (from each of 4 mitral annular sites, inferior, anterior, septum and lateral) were obtained. Mean peak (S) MAM velocity (Sm), mean annular early (D) velocity (Em) by PWDTI and mean mitral annular plane (S) excursion (MAPSE) by MME were calculated by averaging of values measured at each annular site. BNP levels were measured by a rapid immunoassay and blinded to cardiologist making the assessment of LV function. RESULTS: MAPSE < 12 mm determined by MME has 90% sensitivity, 88% specificity & 89% accuracy for detection of LVEF <50%, while these values were 94%, 93% & 94% respectively for (Sm) < 8 cm/s determined by PWDTI. BNP level>75 pg/ml has 98% sensitivity, 90% specificity & 97% accuracy for detection of LV Dys either (S,D, or both). BNP levels were significantly higher in patients with combined (S & D) Dys. Than those with only (S) Dys, the later group had significantly higher BNP levels than those with only (D) Dys. (1054.5 +/- 202.3 pg/ml vs. 500 +/- 39.9 pg/ml & 500 +/- 39.9 pg/ml vs. 215.3 +/- 100.9 pg/ml respectively, P < 0.001) & each were significantly higher than control group (12.3 +/- 5.7 pg/ml, P < 0.001). Significant correlations (P < 0.001 for all) were found between BNP levels and Em (r =-0.82), Sm (r=-0.7), early transmitral (E) to Em ratio (r=0.61), MAPSE (r= 0.54), LVEF(r=-0.64) & LV end D dimension (r=0.63). CONCLUSION: MME and PWDTI used for assessment of MAM are useful methods for evaluation of LV function but parameters measured by PWDTI correlate more strongly with plasma BNP levels than those measured by MME and provide a simple, sensitive, accurate and reproducible tool for early diagnosis of LV dysfunction. PMID- 16046189 TI - What is a "normal" right ventricle? AB - AIMS: The aim of this project was to define normal values of right ventricular (RV) volumes and ejection fraction (EF) in healthy population using 2D echocardiography. METHODS AND RESULTS: The "patient" group comprised 91 healthy volunteers aged 17-62 years. RV volumetry was based on ellipsoidal shell model method. Left ventricular (LV) volumes were assessed by Teichholz formula. All volumes were indexed per m(2) of BSA and the rate distribution of measured and calculated values were evaluated. The normal range of individual parameters was expressed as mean value+/-2 standard deviations (delta). A pair test was used to compare corresponding results of the RV and LV measurements. The regression analysis was used to test the relationship between LV and RV volumes and age. Indexed enddiastolic and endsystolic RV volumes were 79.1+/-29.9ml and 32.6+/ 19.7ml, respectively, EF being 50+/-9.7% in men and 58+/-13.6% in women. No correlation with patient's age was observed. CONCLUSION: Enddiastolic and endsystolic volumes of RV were significantly higher than those of LV. EF of RV was lower as compare to LV. Right ventricular EF in men was lower than that in women. There was no correlation between EF and patient's age. PMID- 16046190 TI - Gliotoxin-induced cytotoxicity in three salmonid cell lines: cell death by apoptosis and necrosis. AB - Epithelial (CHSE-214), fibroblast (RTG-2) and macrophage (RTS11) cell lines from Chinook salmon and rainbow trout were tested for their sensitivity to gliotoxin, a fungal metabolite. Gliotoxin treatment for 6 or 24 h caused cell viability to decrease in a dose-dependent manner, with effective concentrations (EC50s) being similar for the three cell lines but varying with exposure time. Under some exposure conditions, hallmarks of apoptosis were detected. Apoptosis was evaluated by the appearance of fragmented nuclei upon H33258 staining and of genomic DNA laddering into 180 bp oligomers. Gliotoxin induced cell detachment in RTG-2 and CHSE-214 cultures, under some conditions. These were the only cultures of these two cell lines in which apoptosis was detected, and apoptotic cells appeared more frequent in the detached population. At the highest concentration, 15 microM, the cells died by an alternative mode, likely necrosis. By contrast, in RTS11 cultures cell detachment was not observed, and apoptosis occurred over a wider concentration range, even 15 microM, reaching levels of over 90%. The preferential death by necrosis for epithelial cells (CHSE-214) and by apoptosis for macrophages (RTS11) could be a beneficial host response to gliotoxin producing fungi, leading respectively to the development and then resolution of inflammation. PMID- 16046191 TI - Design and rationale of the Utah obesity study. A study to assess morbidity following gastric bypass surgery. AB - PURPOSE: This paper details the design and baseline characteristics of a study on the morbidity associated with Roux-en-Y gastric bypass surgery (GBP) in severely obese adults. This study is designed to assess the effectiveness of GBP in reducing morbidity and maintaining weight loss. A wide array of clinical tests and psycho-behavioral questionnaires are included as part of the study. METHODS: Three groups (n=1156 severely obese) have been recruited for this study: cases who were approved for and participated in surgery (n=415), a control group of GBP seeking individuals who were denied surgery (n=420) and a control group that was randomly chosen from a population of severely obese participants who were not seeking GBP (n=321). Clinical measures include: a physician interview and detailed medical history, resting electro- and echocardiograms, a submaximal exercise treadmill test and electrocardiogram, pulmonary function, limited polysomnography, resting metabolic rate, anthropometrics, resting and exercise blood pressure, comprehensive blood chemistry and urinalysis and dietary, quality of life and physical activity questionnaires. Most participants (76%) were tested following an overnight stay in a clinical research center. Remaining participants underwent less extensive testing in an outpatient clinic. RESULTS: Baseline characteristics of the 1156 participants are available for selected measures. Mean+/-S.D. for BMI was 46+/-7.5 kg/m(2) (range=33 to 92) and for age was 44+/ 11.4 years (range=18 to 72). The prevalence of diabetes and hypertension was 19% and 35%, respectively. Of the participants who had an echocardiogram or polysomnogram, 92% had left-ventricular hypertrophy and 85% had mild to severe sleep apnea. The two control groups were similar to the surgical group. At approximately 24 months, all participants will have a second clinical examination. Statistical comparisons of changes in morbidity variables will be made between the surgical and control groups. CONCLUSIONS: This study design facilitates assessment of risks and benefits of GBP to perform recommendations on whether or not to perform surgery on the severely obese patient. Baseline and 2 year exams provide valuable data for comparison to future long-term follow-up data that can be collected at 5 and 10 years. PMID- 16046192 TI - DNA-arrays with electrical detection: a label-free low cost technology for routine use in life sciences and diagnostics. AB - Fast and highly parallel DNA analysis are essential for improved biomedical research and development. Currently fluorescence-based methods are state of the art in DNA microarray analysis. The necessity to modify the target DNA with labels is costly, laborious and requires skilled personnel. Moreover, false positive calls from unspecific adsorption are possible and it is difficult to discriminate perfect matching target sequences from those with a single mismatch. In this paper a new and simple electrochemical approach for hybridisation detection without the need of labelling the target DNA is described. The EDDA (Electrically Detected Displacement Assay) method uses a solution of short redox labelled signalling oligonucleotides (oligonucleotides carrying a covalently attached redox active compound like ferrocene) to characterize the hybridisation state of label-free capture probe DNA immobilised on gold electrodes. The number of capture probes associated with signalling oligonucleotides is determined by chronocoulometry. This technique allows to separate the electrochemical response of capture probe associated signal probes from the response of freely diffusing signalling probes. In the absence of the complementary target sequences the redox labelled signalling probes at the surface give rise to an instantaneous increase of the detection signal, while freely diffusing signalling probes show a significantly delayed response. Hybridisation with targets complementary to the capture probe displace the loosely associated signalling probes thereby decreasing the instantaneous signal. Besides an introduction to the EDDA technology, data validating the method for biological material will be presented and an outlook to the detection of single nucleotide polymorphisms (SNPs) is given. PMID- 16046193 TI - Comparative TFIIS-mediated transcript cleavage by mammalian RNA polymerase II arrested at a lesion in different transcription systems. AB - Upon prolonged arrest at a cyclobutane pyrimidine dimer (CPD), RNAPII can reverse translocate, misaligning the 3'-end of the RNA from its active site. Transcription factor SII (TFIIS) is required for cleavage of the disengaged 3' end and restoration of its correct positioning. We have previously shown in vitro that when RNAPII is arrested at a CPD, TFIIS-induced cleavage results in shortened transcripts. Here, we hypothesized that the pattern of transcript cleavage does not depend solely upon TFIIS itself, but also on some other general transcription factors (GTFs) and/or their effects on RNAPII. To test this hypothesis we compared three in vitro transcription systems which differ with respect to the mode of initiation and the requirement for GTFs. The first consisted of RNAPII and GTFs from rat liver, and required a eukaryotic promoter for initiation. The other two supported transcription in the absence of any GTFs or promoter sequences. In each case, a CPD on the transcribed strand was a complete block for RNAPII translocation. However, the effect of TFIIS on transcript cleavage varied. In the promoter-initiated system, distinct transcripts up to about 20 nucleotides shorter than the uncleaved original one were produced. In the other two systems, the transcripts were degraded nearly completely. Introduction of GTFs partially interfered with cleavage, but failed to reproduce the pattern of transcript lengths observed with the promoter initiated system. Our results suggest that the extent of TFIIS-mediated transcript cleavage is a well-orchestrated process, depending upon other factors (or their effects on RNAPII), in addition to TFIIS itself. PMID- 16046194 TI - DNA-PK is responsible for enhanced phosphorylation of histone H2AX under hypertonic conditions. AB - Exposure of cells to hypertonic medium after X-irradiation results in a 3-4-fold increase in the phosphorylation of histone H2AX (gammaH2AX) at sites of radiation induced DNA double-strand breaks. This increase was previously associated with salt-induced radiosensitization and inhibition of repair of DNA double-strand breaks. To examine possible mechanisms for the increase in foci size, chemical inhibitors of kinase and phosphatase activity and cell lines deficient in ATM and DNA-PK, two kinases known to phosphorylate H2AX, were examined. H2AX kinase and phosphatase activity were maintained in the presence of high salt. ATM mutant HT144 melanoma cells showed the expected 3-4-fold increase in H2AX phosphorylation in the presence of 0.5M Na(+). However, DNA-PKcs deficient M059J cells failed to respond to hypertonic treatment and M059J Fus1 cells corrected for this deficiency showed the expected increase in foci size. Although the active phosphoform of ATM, phosphoserine-1981, increased after irradiation, the level was unaffected by the addition of 0.5M Na(+). Instead, 0.5M Na(+) caused a partial redistribution of serine-1981-ATM to perinuclear regions. Hypertonic medium added after irradiation was effective in inhibiting rejoining of the radiation-induced double-strand breaks even in DNA-PK deficient M059J cells. We suggest that hypertonic treatment following irradiation inhibits double-strand break rejoining that in turn maintains DNA-PK activity at the site of the break, enhancing the size of the gammaH2AX foci. PMID- 16046195 TI - In vitro evaluation of efficacy of 5 methods of disinfection on mouthpieces and facemasks contaminated by strains of cystic fibrosis patients. AB - INTRODUCTION: Home-nebulizers are a potential source of bacterial infection of the respiratory tract in patients suffering from cystic fibrosis. Recommendations for disinfecting this equipment are often arbitrary and sometimes contradictory. OBJECTIVE: To assess in vitro the effectiveness of 5 methods of disinfecting this equipment. METHODS: 160 mouthpieces and 160 masks of nebulizers were artificially and massively contaminated with 16 strains of germs found in patients with cystic fibrosis (Staphylococcus aureus, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Burkholderia cenocepacia, Alcaligenes xylosoxydans). A controlled comparison was carried out of the five methods of disinfection (hypochlorite solution (0.02% active chlorine), acetic acid 3.5%, Hexanios 0.5%, washing-up detergent 0.5% and a dishwasher), tested with and without drying. Standardised bacteriological sampling took place 4 h after disinfecting. RESULTS: Following treatment, the disappearance of the germ was recorded in 84.1% of cases, and effective disinfecting (reduction>5 log CFU/mL) in another 10.6%. Disinfection failure (5.3%) was found almost only in the case of acetic acid against Staphylococcus aureus. CONCLUSION: With the exception of acetic acid, the methods of disinfecting tested in this study appeared to be effective against common bacterial pathogens in cystic fibrosis. PMID- 16046196 TI - MBL2 polymorphisms screening in a regional Italian CF Center. AB - We performed MBL2 genotyping in 47 CF patients-cared of at the regional CF Centre of Trieste-trying to establish a correlation within allelic variants of MBL2 and modification of patients' clinical outcome. FEV1 values were significantly lowered and a significantly earlier age at onset of Pseudomonas aeruginosa colonisation was found in CF patients with at least one MBL2 variant. PMID- 16046197 TI - Restoration of respiratory muscle function following spinal cord injury. Review of electrical and magnetic stimulation techniques. AB - Respiratory complications are a leading cause of morbidity and mortality in patients with spinal cord injury. Several techniques, currently available or in development, have the capacity to restore respiratory muscle function allowing these patients to live more normal lives and hopefully reduce the incidence of respiratory complications. Bilateral phrenic nerve pacing, a clinically accepted technique to restore inspiratory muscle function, allows patients with ventilator dependent tetraplegia complete freedom from mechanical ventilation. Compared to mechanical ventilation, phrenic nerve pacing provides patients with increased mobility, improved speech, improved comfort level and reduction in health care costs. The results of clinical trials of laparoscopically placed intramuscular diaphragm electrodes suggest that diaphragm pacing can also be achieved without the need for a thoracotomy and associated long hospital stay, and without manipulation of the phrenic nerve which carries a risk of phrenic nerve injury. Other clinical trials are being performed to restore inspiratory intercostal function. In patients with only unilateral phrenic nerve function who are not candidates for phrenic nerve pacing, combined intercostal and unilateral diaphragm pacing appears to provide benefits similar to that of bilateral diaphragm pacing. Clinical trials are also underway to restore expiratory muscle function. Magnetic stimulation, surface stimulation and spinal cord stimulation of the expiratory muscles are promising techniques to restore an effective cough mechanism in this patient population. These techniques hold promise to reduce the incidence of respiratory tract infections, atelectasis and respiratory failure in patients with spinal cord injury and reduce the morbidity and mortality associated with these complications. PMID- 16046198 TI - Development of mechanics and pulmonary reflexes. AB - The mechanical properties of the respiratory system are paramount in converting neural output into ventilation. The highly compliant chest wall of the newborn results in chest distortion and volume loss during inspiration and, as the chest is also unable to resist the inward recoil of the lung, there is a reduction in lung volume at end expiration (functional residual capacity) and a tendency for alveoli to collapse. Vagal innervation of the lungs and airways is responsible for eliciting various reflexes that result in the dynamic modification of respiratory mechanics and an improvement in ventilation. From the first breath, the newborn increases the frequency of augmented breaths to improve lung compliance and prolongs the expiratory time constant in order to increase the amount of air remaining in the lung at end expiration and help prevent lung collapse. This review examines the respiratory mechanics of the mammalian neonate at birth and during early development together with the vagal reflexes that are responsible for the dynamic modification of respiratory mechanics in order to ensure that effective gas exchange occurs from birth. PMID- 16046199 TI - Development and validation of a high performance liquid chromatography assay for 17alpha-methyltestosterone in fish feed. AB - 17alpha-Methyltestosterone (MT) is used to manipulate the gender of a variety of fish species. A high performance liquid chromatography (HPLC) internal standard method for the determination of 17alpha-methyltestosterone in fish feed using 3beta-methoxy-17beta-hydroxyandrost-5-en-7-one as internal standard (IS) has been developed. The method has been validated for the quantitation of MT in fish feed using 245 nm UV absorbance as the parent wavelength and 255 nm as a qualifier wavelength. The method was validated in the concentration range of 15.0-120 mg/kg of 17alpha-methyltestosterone in fish feed. Method was also found to be suitable for other feeds. PMID- 16046200 TI - Enzymatic diagnosis of medium-chain acyl-CoA dehydrogenase deficiency by detecting 2-octenoyl-CoA production using high-performance liquid chromatography: a practical confirmatory test for tandem mass spectrometry newborn screening in Japan. AB - Many of the previously described enzymatic assay methods for the diagnosis of medium-chain acyl-CoA dehydrogenase (MCAD) deficiency have been dependent upon the measurement of radioisotope-labeled co-products or reduction of electron acceptors. We have developed a direct assay method to detect 2-enoyl-CoA production using high-performance liquid chromatography (HPLC). Crude cell lysate prepared from lymphocytes were incubated with n-octanoyl-CoA and ferrocenium hexafluorophosphate. The detection of 2-octenoyl-CoA was significantly reproducible. We applied the assay to samples from four infants suspected to have MCAD deficiency by tandem mass spectrometry (MS/MS) newborn screening conducted in the Hiroshima area of Japan. Three of them were proved to have pathologically reduced residual enzyme activities, although they were associated with various clinical and biochemical phenotypes. In addition, another symptomatic Japanese patient and her presymptomatic sibling who were detected by MS/MS selective screening were successfully diagnosed by our enzymatic assay. These results indicate that the method can be a useful confirmatory test for MS/MS screening of MCAD deficiency. PMID- 16046201 TI - Studies on the metabolism and toxicological detection of the new designer drug 4' methyl-alpha-pyrrolidinobutyrophenone (MPBP) in rat urine using gas chromatography-mass spectrometry. AB - The aim of the presented study was to identify the metabolites of the new designer drug 4'-methyl-alpha-pyrrolidinobutyrophenone (MPBP) in rat urine using GC-MS techniques. After enzymatic hydrolysis, extraction and various derivatizations, seven metabolites of MPBP could be identified suggesting the following metabolic steps: oxidation of the 4'-methyl group to the corresponding alcohol and further oxidation to the respective carboxy compound, hydroxylation of the pyrrolidine ring followed by dehydrogenation to the corresponding lactam or reduction of the keto group to the 1-dihydro compound. A previously published GC-MS-based screening procedure for pyrrolidinophenones involving enzymatic hydrolysis and mixed-mode solid-phase extraction of urine samples allowed detection of MPBP metabolites. Assuming similar metabolism and dosages in humans, an intake of MPBP should be detectable via its metabolites in urine. PMID- 16046202 TI - High-performance liquid chromatographic determination of lamivudine in human serum using liquid-liquid extraction; application to pharmacokinetic studies. AB - A simple, fast, and sensitive high performance liquid chromatographic (HPLC) assay was developed for quantitation of lamivudine in human serum. Lamivudine is polar compound and its extraction from the human serum in previously published HPLC methods involved either protein precipitation or solid phase extraction techniques. However, existence of endogenous peaks which interfere with the drug or appeared as late eluting peaks and lead to long run time of analysis has been reported. Application of either an ion pairing agent in the mobile phase or time consuming column purge has been used in the published methods. Present paper describes liquid - liquid extraction of lamivudine and internal standard (famotidine) using dichloromethane-isopropyl alcohol (1:1, v/v) as an extracting solvent and salting out approach. The mobile phase was a mixture of phosphate buffer (0.05 M) containing triethylamine (1 mL/L, v/v; pH 3.5) and methanol (91:9, v/v) at a flow rate of 2.2 mL/min. The analysis was performed on a column (150 mm x 6 mm i.d.) which was packed with 5 microm particles of ODS packing material. Under these conditions no interference in the assay from any endogenous substance was observed. The limit of quantification was evaluated to be 5 ng/mL. Accuracy and precision of the method were also studied and the technique was shown to be selective and linear into the concentration range of 5-2500 ng/mL. This method has been used in two randomized crossover bioequivalence studies of 100 and 150 mg lamivudine preparations in 12 and 24 healthy volunteers, respectively. PMID- 16046203 TI - Body mass index and health among Union Army veterans: 1891-1905. AB - This paper explores the relationship between BMI and several health conditions among Union Army veterans who had medical examinations between 1891 and 1905. We find that BMI, when used as a proxy of nutrition, helps to explain morbidity and mortality differentials among veterans. There is evidence suggesting that among Union Army veterans extremely low or high BMIs were both associated with poor health, as indicated by a higher level of disability rating, higher risk of developing certain diseases, and higher mortality risk than those associated with having normal weight. Compared to veterans with normal weight, underweight veterans were more likely to be diagnosed with cardiovascular, respiratory and gastrointestinal diseases, but were less likely to be diagnosed with rheumatic and musculo-skeletal conditions at the first examination. High BMI levels are also associated with a higher risk of developing cardiovascular and rheumatic diseases, and higher mortality in the 20 years after the first examination. We performed a longitudinal analysis to study the association between earlier BMI as well as weight change and later development of diseases. The results suggest that, as a predictor of diseases, the explanatory power of BMI becomes lower the farther into the future we try to predict. Compared with those who maintained the same weight, veterans who gained weight were associated with a lower risk of being diagnosed with gastrointestinal diseases at their second examination. PMID- 16046204 TI - Bone as an ion exchange organ: evidence for instantaneous cell-dependent calcium efflux from bone not due to resorption. AB - The current study tests the hypothesis that basal level and minute-by-minute correction of plasma Ca2+ by outward and inward Ca2+ fluxes from and into an exchangeable ionic pool in bone is controlled by an active partition system without contributions from the bone remodeling system. Direct real-time measurements of Ca2+ fluxes were made using the scanning ion-selective electrode technique (SIET) on living bones maintained ex vivo in physiological conditions. SIET three-dimensional measurements of the local Ca2+ concentration gradient (10 microm spatial resolution) were performed on metatarsal bones of weanling mice after drilling a 100-mum hole through the cortex to expose the internal bone extracellular fluid (BECF) to the bathing solution, whose composition mimicked the extracellular fluid (ECF). Influxes of Ca2+ towards the center of the cortical hole (15.1+/-4.2 pmol cm-2 s-1) were found in the ECF and were reversed to effluxes (7.4+/-2.9 pmol cm-2 s-1) when calcium was depleted from the ECF, mimicking a plasma demand. The reversal from influx to efflux and vice versa was immediate and fluxes in both directions were steady throughout the experimental time (>or=2 h, n=14). Only the efflux was nullified within 10 min by the addition of 10 mM/L Na-Cyanide (n=7), demonstrating its cell dependence. The timeframes of the exchanges and the stability of the Ca2+ fluxes over time suggest the existence of an exchangeable calcium pool in bone. The calcium efflux dependency on viable cells suggests that an active partition system might play a central role in the short-term error correction of plasma calcium without the contribution of bone remodeling. PMID- 16046205 TI - Biological properties and mechanism of action of ibandronate: application to the treatment of osteoporosis. AB - Bisphosphonates, with their proven efficacy and safety, are the most commonly prescribed treatment for women with postmenopausal osteoporosis; however, optimal efficacy is often not achieved due to poor patient adherence to medication. Poor adherence leads to an increased risk of fracture, which itself results in morbidity, elevated healthcare costs and potentially, mortality. Although weekly rather than daily dosing of bisphosphonates has improved adherence, there remains a significant problem, and dosing less frequently than weekly has been suggested as a possible means for further improving adherence. Ibandronate is a new bisphosphonate that has a specific structure and set of characteristics that enable less frequent dosing than currently available bisphosphonates. This review provides details of the general structural features of all bisphosphonates and how these are understood to contribute to their functions in osteoporosis treatment. From this, the unique structure of ibandronate is described, along with how this translates into the high antiresorptive potency, favorable bone binding, persistence in bone, and good tolerability that permits less frequent dosing. Finally, the clinical evidence for ibandronate is briefly presented, demonstrating the viability of less frequent dosing, with its potential benefits for patient convenience and adherence to therapy. PMID- 16046206 TI - Novel insights into actions of bisphosphonates on bone: differences in interactions with hydroxyapatite. AB - Bisphosphonates are now the most widely used drugs for diseases associated with increased bone resorption, such as osteoporosis. Although bisphosphonates act directly on osteoclasts, and interfere with specific biochemical processes such as protein prenylation, their ability to adsorb to bone mineral also contributes to their potency and duration of action. The aim of the present study was to compare the binding affinities for hydroxyapatite (HAP) of 6 bisphosphonates currently used clinically and to determine the effects of these bisphosphonates on other mineral surface properties including zeta potential and interfacial tension. Affinity constants (K(L)) for the adsorption of bisphosphonates were calculated from kinetic studies on HAP crystal growth using a constant composition method at 37 degrees C and at physiological ionic strength (0.15 M). Under conditions likely to simulate bisphosphonate binding onto bone, there were significant differences in K(L) among the bisphosphonates for HAP growth (pH 7.4) with a rank order of zoledronate > alendronate > ibandronate > risedronate > etidronate > clodronate. The measurements of zeta potential show that the crystal surface is modified by the adsorption of bisphosphonates in a manner best explained by molecular charges related to the protonation of their side-chain moieties, with risedronate showing substantial differences from alendronate, ibandronate, and zoledronate. The studies of the solid/liquid interfacial properties show additional differences among the bisphosphonates that may influence their mechanisms for binding and inhibiting crystal growth and dissolution. The observed differences in kinetic binding affinities, HAP zeta potentials, and interfacial tension are likely to contribute to the biological properties of the various bisphosphonates. In particular, these binding properties may contribute to differences in uptake and persistence in bone and the reversibility of effects. These properties, therefore, have potential clinical implications that may be important in understanding differences among potent bisphosphonates, such as the apparently more prolonged duration of action of alendronate and zoledronate compared with the more readily reversible effects of etidronate and risedronate. PMID- 16046207 TI - Bone graft versus BMP-7 in a critical size defect--cranioplasty in a growing infant model. AB - Little data are available as regards to the action of bone morphogenetic protein 7 (rhBMP-7) in growing organisms. We put forward two hypotheses: Firstly, that regeneration of calvarial defects with autologous bone grafts would result in equivalent volume and shape as compared to calvaria regenerated with BMP-7. Secondly, that cranial development would remain undisturbed in infant individuals. A one-sided defect of the parietal bone (2x4 cm) including the coronal suture was generated in 2-month-old minipigs (n=17). Group 1: no further treatment (n=5); group 2: particulated iliac bone graft (n=6); group 3: rhBMP-7 composite (500 microg/g collagen+Carboxymethylcellulose, n=6). After the experimental period (4 months) with fluorochrome labeling, examination was performed by computed-tomography and non-decalcified histology. Group 1: major bony gaps remained, proving that defects of critical size were generated. Group 2: minor bony gaps remained, the bone volume was significantly reduced on the treated as compared to untreated sides (P=0.028). Group 3: bony continuity was seen in all cases and no significant difference of bone volumes of treated versus untreated sides (P=0.075) was found. Skull diameters increased by 16.4% but the physiological centrifugal cranial expansion remained undisturbed. Our first hypothesis was contradicted: contrary to our former assumption, bone induction by rhBMP-7 was superior to particulated bone transplants. In this growing model, calvaria approaching normal volume and shape were observed. However, only the quantity not the quality of bone regenerates was different. Our second hypothesis was confirmed: disruption of further cranial development was not seen after bone transplantation or rhBMP-7 implantation. PMID- 16046208 TI - Inhibition of cortical and cancellous bone formation in glucocorticoid-treated OVX sheep. AB - INTRODUCTION: Little is known about the effects of steroids on cortical bone. The purpose of this study was to explore the effects of glucocorticoid treatment on cortical bone density, strength, and formation of new bone in ovariectomized sheep and to compare it to cancellous bone. MATERIALS AND METHODS: Sixteen ovariectomized merino sheep either received a 6-month glucocorticoid treatment (GLU; 0.45 mg/kg/day Methylprednisolone s.c.) or were left untreated (control). After 2 and 4 months, newly formed bone was labeled by the administration of calceingreen and Tetracycline-hydrochloride. After 12 months, the animals were sacrificed and cortical specimens were obtained from the tibial diaphysis. Cancellous bone specimens were harvested from the proximal tibia. All specimens were scanned for apparent BMD by pQCT and tested mechanically. Formation of new bone was analyzed using histological slices of the femoral condyles and cross sections of the mid-diaphysis of the tibia. RESULTS: The intracortical formation of new bone in glucocorticoid-treated sheep was 70% lower after 2 months and 80% lower after 4 months. Six months after the termination of the steroid administration, the active bone area was 20% lower than in the control group. Cortical width and cortical bone area were reduced by 7-8% and marrow area increased by 8% in steroid-treated animals compared to control animals. Neither cortical apparent bone mineral density nor biomechanical parameters of cortical bone specimens differed between the groups. Cancellous bone formation in steroid treated animals declined by 68% after 2 months and by 90% after 4 months. After 1 year, cancellous bone formation was reduced by 38% compared to control. The apparent cancellous bone mineral density and cancellous bone compressive stiffness were reduced by 34% and 55%, respectively. CONCLUSION: A six-month glucocorticoid treatment of ovariectomized sheep resulted in a substantial reduction of bone formation both in cortical and in cancellous bone and reduced cortical width by increased endosteal resorption. Beyond changes in cancellous bone, impaired cortical bone remodeling may add to the increased fracture risk at the hip or the distal radius in patients treated with steroids. PMID- 16046209 TI - Polychrome labeling of bone with seven different fluorochromes: enhancing fluorochrome discrimination by spectral image analysis. AB - Bone formation and remodeling in vivo can be assessed by polychrome labeling using calcium-binding fluorescent dyes. The number of fluorochromes, however, limits this technique due to the fact that with increasing number, fluorescent spectra inevitably overlap, which makes discrimination more difficult. In order to enhance discrimination, we performed spectral image analysis. Non-critical size defects of the femur of male Wistar rats served as a model for bone formation. Eight different fluorochromes (calcein blue, xylenol orange, calcein, alizarine complexone, doxycycline, rolitetracycline, hematoporphyrin, and BAPTA) were administered sequentially subcutaneously every third day starting at day 4 after surgery. Following, bone specimen were embedded in methylmethacrylate and analyzed by spectral image acquisition using a Sagnac type interferometer (ASI, Israel). Seven of the eight applied fluorochromes could be resolved using spectral image examination. With BAPTA, we present a new fluorochrome suitable for bone labeling. Due to the superior sensitivity of the spectral image acquisition, the thickness of the bone sections could be reduced so that 5 mum thick sections could be analyzed. Spectral decomposition and subsequent linear unmixing allows depiction of each individual fluorochrome without interference of any other, enabling a reliable and superior morphometric analysis of labeled regions. PMID- 16046210 TI - Sex-specific quantitative trait loci contribute to normal variation in bone structure at the proximal femur in men. AB - Bone structure is an important determinant of osteoporotic fracture. In women, bone structure is highly heritable, and several quantitative trait loci (QTL) have been reported. There are few comparable data in men. This study in men aimed at establishing the heritability of bone structure at the proximal femur, identifying QTL contributing to normal variation in bone structure, and determining which QTL might be sex-specific. Bone structure at the proximal femur was measured in 205 pairs of brothers age 18-61. Heritability was calculated, and linkage analysis performed on phenotypes at the proximal femur. Heritability estimates ranged from 0.99 to 0.39. A genome wide scan identified suggestive QTL (LOD>2.2) for femoral shaft width on chromosome 14q (LOD=2.69 at position 99 cM), calcar femorale at chromosome 2p (LOD=3.97 at position 194 cM) and at the X chromosome (LOD=3.01 at position 77 cM), femoral neck width on chromosome 5p (LOD=2.28 at position 0 cM), femoral head width on chromosome 11q (LOD=2.30 at position 131 cM) and 15q (LOD=3.11 at position 90 cM), and pelvic axis length on chromosome 4q (LOD=4.16 at position 99 cM) and 17q (LOD=2.80 at position 112 cM). Comparison with published data in 437 pairs of premenopausal sisters from the same geographical region suggested that 3 of the 7 autosomal QTL were male specific. This study demonstrates that bone structure at the proximal femur in healthy men is highly heritable. The occurrence of sex-specific genes in humans for bone structure has important implications for the pathogenesis and treatment of osteoporosis. PMID- 16046211 TI - Implicit and explicit appraisals of the importance of intrusive thoughts. AB - To evaluate cognitive theories of obsessions, the current study experimentally manipulated appraisals of the importance of intrusive thoughts. Undergraduate students (N = 156) completed measures of obsessive-compulsive disorder (OCD) symptoms and beliefs and were primed with a list of commonly reported unwanted thoughts. Participants were then informed that unwanted thoughts are either (1) significant and indicative of their personal values, or (2) meaningless, or participants (3) received no instructions about unwanted thoughts. Participants then completed implicit and explicit measures of self-evaluation and interpretations of their unwanted thoughts. Results indicated that the manipulation shifted implicit appraisals of unwanted thoughts in the expected direction, but not self-evaluations of morality or dangerousness. Interestingly, explicit self-esteem and beliefs about the significance of unwanted thoughts were associated with measures of OCD beliefs, whereas implicit self-evaluations of dangerousness were better predicted by the interaction of pre-existing OCD beliefs with the manipulation. Results are discussed in terms of divergent predictors of implicit and explicit responses to unwanted thoughts. PMID- 16046212 TI - Role of benoxaprofen and flunoxaprofen acyl glucuronides in covalent binding to rat plasma and liver proteins in vivo. AB - Benoxaprofen (BNX) has been implicated in rare but serious hepatotoxicity which led to its withdrawal from the world market. Flunoxaprofen (FLX), a structural analog, appears to be less toxic. It has been postulated that the nonsteroidal antiinflammatory drugs associated toxicity may be related to covalent modification of proteins by their reactive acyl glucuronides, and the extent of covalent protein binding depends on both reactivity of the acyl glucuronide and the exposure to the reactive metabolite. The disposition of BNX and FLX in rats were compared upon intravenous administration of 20 mg/kg of BNX, FLX or their metabolites. Covalent binding of BNX and FLX to plasma and liver proteins were also determined, and an immunochemical approach was used to detect their hepatic targets. Similar concentrations of plasma protein adducts for BNX and FLX were detected even though the AUC of BNX-glucuronide (BNX-G) was almost twice that of FLX-glucuronide (FLX-G). Similar concentrations of liver protein adducts for BNX and FLX were also detected at 8 h, however, the hepatobiliary exposure of BNX-G was only 1/3rd that of FLX-G indicating that BNX-G was more reactive than FLX-G, which was in agreement with in vitro data. Proteins of 110 and 70 kDa were the major liver protein targets modified by covalent attachment of BNX and FLX. In conclusion, measuring covalent binding to tissue proteins in animals in addition to plasma adducts should be considered when evaluating and comparing carboxylic acid analogs that form reactive acyl glucuronides. PMID- 16046213 TI - Activation of the aryl hydrocarbon receptor by berberine in HepG2 and H4IIE cells: Biphasic effect on CYP1A1. AB - Berberine has long been considered a candidate for an antimalarial drug. It exerts a plethora of biological activities and has been used in the treatment of diarrhea and gastro-enteritis for centuries. Here we provide evidence that berberine activates the aryl hydrocarbon receptor (AhR) in human hepatoma (HepG2) and rat hepatoma cells stably transfected with a dioxin responsive element fused to the luciferase gene (H4IIE.luc). AhR was activated by high doses of berberine (10-50 microM) after 6 and 24 h of incubation as revealed by CYP1A1 mRNA expression (HepG2) and AhR-dependent luciferase activity (H4IIE.luc). Berberine induced nuclear translocation of AhR-GFP chimera transiently transfected to Hepa1c1c7 cells. In contrast, low doses of berberine (<1 microM) and prolonged times of the treatments (48 h) failed to produce any activation of AhR in H4IIE.luc cell line. HPLC analysis ruled out the hypothesis that the loss of berberine capacity to activate AhR in H4IIE.luc cells is due to metabolic inactivation of the alkaloid. We demonstrate that berberine is a potent inhibitor (IC50=2.5 microM) of CYP1A1 catalytic activity (EROD) in HepG2 cell culture and in recombinant CYP1A1 protein. Collectively, our results imply that while berberine activates the Ah receptor, it is accompanied by inactivation of the catalytic activity of CYP1A1 and occurs at concentrations that exceed those predicted to occur in vivo. Given these data, it appears that activation of the AhR pathway by berberine has a low toxicological potential. PMID- 16046214 TI - Oxidized LDL induces a coordinated up-regulation of the glutathione and thioredoxin systems in human macrophages. AB - Using DNA microarray analysis, we found that human macrophages respond to oxidized low-density lipoprotein (oxLDL) by activating the antioxidative glutathione and thioredoxin systems. Several genes of the glutathione and thioredoxin systems were expressed at high levels in macrophages when compared to 80 other human tissues and cell types, indicating that these systems may be of particular importance in macrophages. The up-regulation of three genes in these systems, thioredoxin (P < 0.005), thioredoxin reductase 1 (P < 0.001) and glutathione reductase (P < 0.001) was verified with real-time RT-PCR, using human macrophages from 10 healthy donors. To investigate the possible role of these antioxidative systems in the development of atherosclerosis, expression levels in macrophages from 15 subjects with atherosclerosis (12 men, 3 women) and 15 matched controls (12 men, 3 women) were analyzed using DNA microarrays. Two genes in the glutathione system Mn superoxide dismutase (P < 0.05) and catalase (P < 0.05) differed in expression between the groups. We conclude that macrophage uptake of oxidized LDL induces a coordinated up-regulation of genes of the glutathione and thioredoxin systems, suggesting that these systems may participate in the cellular defense against oxidized LDL and possibly modulate the development of atherosclerosis. PMID- 16046215 TI - Pleiotropic QTL on chromosome 19q13 for triglycerides and adiposity: the HERITAGE Family Study. AB - Motivated by strong correlations between plasma levels of triglycerides (TG) and adiposity traits, we conducted a series of bivariate genome-wide linkage analyses of TG with body mass index (BMI), total fat mass (FAT), percentage of body fat (FATPC), and abdominal subcutaneous fat (ASF). Maximum lod scores of 3.3, 3.0, 2.2 and 2.4, respectively, were found on chromosome 19q13. This linkage region includes the APOE gene, a predictor of variation in lipid-lipoprotein levels, and the hormone-sensitive lipase (LIPE) gene, a key enzyme in the mobilization of fatty acids from triglyceride stores. In addition, the adiposity measures together with the APOE marker showed significant association with TG levels (p = 0.02 to p = 0.03). In summary, these results suggest that one or more QTLs in the 19q13 region jointly influence TG levels and adiposity. Polymorphisms in the APOE gene, and possibly LIPE gene, appear to be strong candidates for the source of this pleiotropic QTL. PMID- 16046216 TI - Cigarette smoking, regular exercise, and peripheral blood flow. AB - Acute and chronic smoking reduces peripheral blood flow and shear stress, contributing to the increased incidence of peripheral arterial disease in smokers. Currently, it is not known whether physical activity status influences peripheral blood flow among chronic smokers. Blood flow was measured using Doppler ultrasound on the common femoral artery in nine young otherwise healthy sedentary smokers (eight males/one female) and nine physically-active smokers (six males/three females). Physically-active smokers performed strenuous exercise 4.4 times/week for 8 h/week. No significant differences in body fat, blood pressure, and total cholesterol were observed between groups. Basal femoral artery blood flow was approximately 50% higher in physically-active smokers compared with sedentary smokers (259+/-108 ml min(-1) versus 173+/-47 ml min(-1), P<0.05). The higher basal femoral artery blood flow in physically-active smokers compared with sedentary smokers was associated with a 47% higher femoral artery vascular conductance (2.99+/-1.2 U versus 2.03+/-0.5 U, P<0.05) and a 39% lower vascular resistance (0.38+/-0.13 U versus 0.53+/-0.15 U, P<0.05). Cardiac output, systemic vascular resistance, femoral intima-media thickness, and plasma norepinephrine concentration were not different between the groups. We concluded that smokers who habitually perform physical activity demonstrate greater levels of peripheral blood flow and peripheral vascular conductance. The findings from the present cross-sectional study suggest that chronic smokers may be able to negate, at least in part, the adverse effects of chronic smoking on the peripheral vasculature by performing regular physical activity. PMID- 16046217 TI - Asymmetric dimethyl arginine levels correlate with cardiovascular risk factors in patients with erectile dysfunction. AB - BACKGROUND: Erectile dysfunction is related to penile arterial endothelial nitric oxide production. Asymmetric dimethylarginine (ADMA) and E-selectin are often considered plasma markers of endothelial function. OBJECTIVE: This study investigated the relationship between these plasma markers and cardiovascular risk factors in patients with erectile dysfunction. METHODS AND RESULTS: Cardiovascular risk factors, ADMA and E-selectin were assessed in 45 patients with erectile dysfunction. Plasma markers showed associations with baseline risk factors. E-selectin levels showed an inverse relationship with age (p = 0.005) and statin therapy (p = 0.03) and a weak association with concomitant beta blocker therapy (p = 0.05). Compared to these relatively weak associations with cardiovascular risk factors, ADMA levels showed strong associations with pulse pressure (p < 0.001), lack of smoking (p = 0.002) and lipoprotein (a) (p = 0.004) concentrations and weak associations with LDL-cholesterol (p = 0.02), and C reactive protein levels (p = 0.04). ADMA levels correlated with E-selectin (partial r = 0.76; p < 0.001) after adjustment for lipoprotein (a), pulse pressure and smoking. No change in E-selectin or ADMA levels was seen after 70 days therapy with sildenafil and no relationship was found between either plasma marker and the acute pulse wave response to a single challenge dose of sildenafil. CONCLUSION: ADMA levels correlate at baseline with some cardiovascular risk factors including inflammatory markers and lipoprotein (a) in patients with erectile dysfunction. PMID- 16046218 TI - Preoperative antithrombin III activity predicts outcome after surgical repair of acute type A aortic dissection. AB - BACKGROUND: Acute Stanford type A aortic dissection is associated with substantial perioperative morbidity and mortality. A sepsis-like state may lead to antithrombin (AT) III consumption and deficiency. The impact of preoperative AT III activity on outcome in patients undergoing emergency surgery is yet unknown. METHODS: We measured preoperative AT III activity in 99 consecutive patients undergoing emergency aortic surgery for Stanford type A aortic dissection during a 4-year period in a retrospective study. Cardiovascular co morbidities, risk factors and surgical data were recorded and patients were followed for 30-day mortality, and occurrence of multiple organ failure (MOF). RESULTS: During the first 30 days, 15 patients (15%) died, and 8 patients (8%) had MOF. Median AT III levels (IQR) in 30-day non-survivors versus survivors were 64% (52-72) versus 90% (75-97) (p<0.001), and in patients with versus without MOF were 66% (52.3-77.3) versus 88% (72-96) (p=0.018), respectively. Adjusted odds ratios for 30-day mortality and MOF for AT III activity (per % increments) were 0.92 (p=0.007), and 0.96 (p=0.012), respectively, indicating a significant inverse relationship between AT III activity and outcome. CONCLUSION: There is a strong inverse association between preoperative AT III activity and adverse outcome in patients undergoing surgical repair of acute Stanford type A aortic dissection. Larger studies are necessary to determine a cut-off value for AT III and to assess whether patients with low AT III levels benefit targeted therapeutic interventions. PMID- 16046219 TI - Associations of leisure time physical activity, self-rated physical fitness, and estimated aerobic fitness with serum C-reactive protein among 3,803 adults. AB - OBJECTIVE: Serum C-reactive protein (CRP), a marker of systemic inflammation, is a risk factor for cardiovascular disease. Obesity and physical activity are associated with CRP, though population studies are sparse. METHODS AND RESULTS: We assessed the cross-sectional relationship of physical activity, self-rated fitness and estimated aerobic fitness with serum CRP among a population-based random sample of 3,803 Finnish adults aged 25-74 years. Conditioning, commuting and non-conditioning physical activity was measured by a recall and self-rated physical fitness by a questionnaire. A non-exercise test was used to estimate aerobic fitness. The combined association of aerobic fitness and waist-to-hip ratio (WHR) on CRP was also assessed. Self-rated fitness and aerobic fitness had inverse associations with CRP (adjusted for age, WHR, smoking, anti-hypertensive drugs, aspirin, diabetes, hypercholesterolemia, hormone replacement therapy and menopausal status) in both sexes (p < 0.001). An inverse age-adjusted association of conditioning and non-conditioning physical activity with CRP was found in both sexes. After further adjustments, associations remained significant in women for conditioning and non-conditioning activity (p < 0.001 and p = 0.010, respectively) and borderline significant in men (p = 0.074 and p = 0.070, respectively). Commuting physical activity was inversely associated with CRP among women only (p = 0.012). CONCLUSIONS: Physical activity and fitness may have an anti-inflammatory effect, which is independent of obesity. PMID- 16046220 TI - Heart valve calcification in young patients with systemic lupus erythematosus: a window to premature atherosclerotic vascular morbidity and a risk factor for all cause mortality. AB - The objective of the study was to evaluate the association between heart valve calcification and atherosclerosis and outcome in systemic lupus erythematosus (SLE). One-hundred and seven patients with SLE (mean age 45.9 +/- 14.7 years) were studied by 2D transthoracic echocardiography. Mitral annulus calcification (MAC) was detected in 24 patients (22.6%) and aortic valve calcification (AVC) in 22 (20.1%). Both MAC and AVC were associated with older age (r = 0.2, p = 0.02; r = 0.40, p T in the apolipoprotein A5 gene is associated with an increased risk of coronary artery disease and altered triglyceride levels in a Chinese population. AB - Elevation in plasma triglycerides (TG) has been widely accepted as a coronary artery disease (CAD) risk predictor. Recently, a new apolipoprotein playing an important role in TG metabolism named apolipoprotein AV (apoAV) was discovered, which is encoded by the APOA5 gene. Several single nucleotide polymorphisms (SNPs) of APOA5 associated with increased TG concentrations have been identified. We here report that a recently identified genetic variant, c.553G>T in the APOA5 gene which causes a substitution of a cysteine for a glycine residue at amino acid residue 185(G185C) is also associated with increased TG levels. To investigate the association between this genetic variation and the risk of CAD, a case-control study comprising 232 patients with CAD and 302 controls from the same area of China was performed. The minor allele frequencies of c.553G > T for the CAD and control groups were 7.76 and 3.97%, respectively (P = 0.008). In both the CAD and control groups, the T allele carriers had higher serum TG levels than homozygous carriers of the major G allele (CAD group: 2.67 +/- 1.48 mmol/l versus 1.95 +/- 1.02 mmol/l, P = 0.021; controls: 2.31 +/- 1.20 mmol/l versus 1.68 +/- 0.95 mmol/l, P = 0.002). After adjustment for age, gender, body mass index, smoking status, glucose and presence of hypertension, the odds ratio (OR) for CAD in the T allele carriers was 2.089 (95% CI = 1.140-3.830, P = 0.017), in comparison to the individuals without the T allele. These results suggest that the APOA5 c.553G > T polymorphism is an important predictor for hypertriglyceridemia and CAD. PMID- 16046222 TI - Plasma cystatin-C and development of coronary heart disease: The PRIME Study. AB - The pathogenesis of ischemic coronary events involves degradation of the extracellular matrix in atherosclerotic lesions. The cysteine protease inhibitor cystatin-C may be involved in this phenomenon. The association of plasma cystatin C with the incidence of myocardial infarction-coronary death and angina, was examined in a nested case-control (two controls per case) design within the prospective cohort study (Prospective Epidemiological Study of Myocardial Infarction (PRIME Study)) which included 9,758 men aged 50-59 years who were free of coronary heart disease (CHD) on entry and followed for a 5-year period. Three hundred and thirteen participants suffered myocardial infarction or coronary death (n = 159) or angina pectoris (n = 154) during follow-up. Cystatin-C was positively correlated with body mass index (BMI), low-density lipoprotein (LDL) cholesterol, triglycerides and several inflammatory markers such as fibrinogen (r = 0.18), C-reactive protein (CRP) (r = 0.24), interleukin-6 (= 0.20), tumor necrosis factor-alpha (TNFalpha) (r = 0.27) and two TNFalpha receptors: TNFR1A (r = 0.43) and TNFR1B (r = 0.41); and negatively with high-density lipoprotein (HDL) cholesterol (r = -0.25). After adjustment for traditional risk factors (age, diabetes, smoking, hypertension, BMI, triglycerides, LDL- and HDL-cholesterol), cystatin-C was significantly associated with the occurrence of the first ischemic coronary event. However, this association was no longer significant when CRP was included in the analysis. A decrease in glomerular filtration rate did not explain higher cystatin-C in cases than in controls. Cystatin-C appears to participate in the inflammatory phenomenon observed in the atherosclerotic process. Cystatin-C is not a more predictive risk marker of CHD than CRP or interleukin-6, but could be useful in detecting moderate chronic renal disease. PMID- 16046223 TI - Muscle contributions to support during gait in an individual with post-stroke hemiparesis. AB - Walking requires coordination of muscles to support the body during single stance. Impaired ability to coordinate muscles following stroke frequently compromises walking performance and results in extremely low walking speeds. Slow gait in post-stroke hemiparesis is further complicated by asymmetries in lower limb muscle excitations. The objectives of the current study were: (1) to compare the muscle coordination patterns of an individual with flexed stance limb posture secondary to post-stroke hemiparesis with that of healthy adults walking very slowly, and (2) to identify how paretic and non-paretic muscles provide support of the body center of mass in this individual. Simulations were generated based on the kinematics and kinetics of a stroke survivor walking at his self-selected speed (0.3 m/s) and of three speed-matched, healthy older individuals. For each simulation, muscle forces were perturbed to determine the muscles contributing most to body weight support (i.e., height of the center of mass during midstance). Differences in muscle excitations and midstance body configuration caused paretic and non-paretic ankle plantarflexors to contribute less to midstance support than in healthy slow gait. Excitation of paretic ankle dorsiflexors and knee flexors during stance opposed support and necessitated compensation by knee and hip extensors. During gait for an individual with post stroke hemiparesis, adequate body weight support is provided via reorganized muscle coordination patterns of the paretic and non-paretic lower limbs relative to healthy slow gait. PMID- 16046224 TI - A branching process, its application in biology: influence of demographic parameters on the social structure in mammal groups. AB - Branching processes are widely used in biology. This theoretical tool is used in cell dynamics, epidemics and population dynamics. In population dynamics, branching processes are mainly used to access extinction probabilities of populations, groups or families, with the Galton-Watson branching process. Many mammal species live in socially-structured groups, and the smallest units of these groups are lineages (or families) of kin-related individuals. In many primate species, these lineages are matrilines, as females remain in their natal groups most of the time, whereas males generally disperse. Lineage parameters, such as numbers of matrilines, size of each matriline and average degree of relatedness, could strongly influence the genetic composition of groups. Evidence indicates that division along matrilines could induce substantial differentiation among fission groups. Here, we develop a novel mathematical model based on the branching process theory describing demographic dynamics of groups. The main result of this model is an explicit analytical expression of the joint distribution of numbers of lineages and sizes of socially-structured groups. We investigated the influence of parameters such as natality and mortality on the outcome of the process, including extinction probability. Finally, we discuss this theoretical result with respect to biological significance. PMID- 16046225 TI - Optimization of inclusive fitness. AB - The first fully explicit argument is given that broadly supports a widespread belief among whole-organism biologists that natural selection tends to lead to organisms acting as if maximizing their inclusive fitness. The use of optimization programs permits a clear statement of what this belief should be understood to mean, in contradistinction to the common mathematical presumption that it should be formalized as some kind of Lyapunov or even potential function. The argument reveals new details and uncovers latent assumptions. A very general genetic architecture is allowed, and there is arbitrary uncertainty. However, frequency dependence of fitnesses is not permitted. The logic of inclusive fitness immediately draws together various kinds of intra-genomic conflict, and the concept of 'p-family' is introduced. Inclusive fitness is thus incorporated into the formal Darwinism project, which aims to link the mathematics of motion (difference and differential equations) used to describe gene frequency trajectories with the mathematics of optimization used to describe purpose and design. Important questions remain to be answered in the fundamental theory of inclusive fitness. PMID- 16046226 TI - The role of p38 MAP kinase and c-Jun N-terminal protein kinase signaling in the differentiation and apoptosis of immortalized neural stem cells. AB - The two distinct members of the mitogen-activated protein (MAP) kinase family c Jun N-terminal protein kinase (JNK) and p38 MAP kinase, play an important role in central nervous system (CNS) development and differentiation. However, their role and functions are not completely understood in CNS. To facilitate in vitro study, we have established an immortal stem cell line using SV40 from fetal rat embryonic day 17. In these cells, MAP kinase inhibitors (SP600125, SB202190, and PD98059) were treated for 1, 24, 48, and 72 h to examine the roles of protein kinases. Early inhibition of JNK did not alter phenotypic or morphological changes of immortalized cells, however overexpression of Bax and decrease of phosphorylated AKT was observed. The prolonged inhibition of JNK induced polyploidization of immortalized cells, and resulted in differentiation and inhibition of cell proliferation. Moreover, JNK and p38 MAP kinase but not ERK1/2 was activated, and p21, p53, and Bax were overexpressed by prolonged inhibition of JNK. These results indicate that JNK and p38 MAP kinase could play dual roles on cell survival and apoptosis. Furthermore, this established cell line could facilitate study of the role of JNK and p38 MAP kinase on CNS development or differentiation/apoptosis. PMID- 16046227 TI - Combined effects of mechanical and ischemic injury to cortical cells: secondary ischemia increases damage and decreases effects of neuroprotective agents. AB - Traumatic brain injury (TBI) involves direct mechanical damage, which may be aggravated by secondary insults such as ischemia. We utilized an in vitro model of stretch-induced injury to investigate the effects of mechanical and combined mechanical/ischemic insults to cultured mouse cortical cells. Stretch injury alone caused significant neuronal loss and increased uptake of the dye, propidium iodide, suggesting cellular membrane damage to both glia and neurons. Exposure of cultures to ischemic conditions for 24h, or a combination of stretch and 24h of ischemia, caused greater neuronal loss compared to stretch injury alone. Next, we tested the neuroprotective effects of superoxide dismutase (SOD), and the nitric oxide (NO) synthase inhibitors 7-nitroindazole (7-NINA) and lubeluzole. In general, these agents decreased neuronal loss following stretch injury alone, but were relatively ineffective against the combined injury paradigm. A combination of SOD with 7-NINA or lubeluzole offered no additional protection than single drug treatment against stretch alone or combined injury. These results suggest that the effects of primary mechanical damage and secondary ischemia to cortical neurons are cumulative, and drugs that scavenge superoxide or reduce NO production may not be effective for treating the secondary ischemia that often accompanies TBI. PMID- 16046228 TI - Testing for a deficit in single-case studies: effects of departures from normality. AB - In neuropsychological single-case research inferences concerning a patient's cognitive status are often based on referring the patient's test score to those obtained from a modestly sized control sample. Two methods of testing for a deficit (z and a method proposed by Crawford and Howell [Crawford, J. R. & Howell, D. C. (1998). Comparing an individual's test score against norms derived from small samples. The Clinical Neuropsychologist, 12, 482-486]) both assume the control distribution is normal but this assumption will often be violated in practice. Monte Carlo simulation was employed to study the effects of leptokurtosis and the combination of skew and leptokurtosis on the Type I error rates for these two methods. For Crawford and Howell's method, leptokurtosis produced only a modest inflation of the Type I error rate when the control sample N was small-to-modest in size and error rates were lower than the specified rates at larger N. In contrast, the combination of leptokurtosis and skew produced marked inflation of error rates for small Ns. With a specified error rate of 5%, actual error rates as high as 14.31% and 9.96% were observed for z and Crawford and Howell's method respectively. Potential solutions to the problem of non normal data are evaluated. PMID- 16046229 TI - UV inactivation of adenovirus type 41 measured by cell culture mRNA RT-PCR. AB - Adenoviruses are among the most resistant waterborne pathogens to UV disinfection, yet of the 51 serologically distinct human adenoviruses, only a few have been evaluated for their sensitivities to UV irradiation. Human enteric adenoviruses (Ad40 and Ad41) are difficult to cultivate and reliably assay for infectivity, requiring weeks to obtain cytopathogenic effects (CPE). Inoculated cell cultures often deteriorate before the appearance of distinctive CPE making it difficult to obtain reliable and reproducible data regarding UV inactivation. Adenovirus is a double-stranded DNA virus and produces messenger RNA (mRNA) during replication in host cells. The presence of viral mRNA in host cells is definitive evidence of infection. We recently developed a rapid and reliable cell culture-mRNA RT-PCR assay to detect and quantify adenovirus infectivity. Viral mRNA recovered from cell cultures 5-7 days after infection was purified on oligo dT latex, treated with DNase, and amplified by RT-PCR using the primers specific for a conserved region of the hexon late mRNA transcript. Treatment of approximately 10(4) Ad41 with different doses of 254 nm germicidal UV radiation resulted in a dose-dependent loss of infectivity. As UV doses were increased from 75 to 200 mJ/cm2, virus survival decreased and no virus infectivity (measured by detectable mRNA) was found at a dose of 225 mJ/cm2 or higher. Our results using the cell culture mRNA RT-PCR assay indicate that Ad41 is more resistant to UV radiation than in a previous study using a conventional cell culture infectivity assay. Results were more similar to those found for Ad 40 using CPE as a measure of infectivity in another previous study. PMID- 16046230 TI - Inhibition of dentine demineralization by zinc oxide: in vitro and in situ studies. AB - OBJECTIVE: The objective of this study was to evaluate the inhibitory effect of zinc oxide on dentine demineralization in vitro and in situ. METHODS: In the in vitro study, dentine specimens treated with a zinc oxide suspension were demineralized in a pH 5 solution. In the in situ study, subjects wore dentine specimens on their teeth and instructed to rinse with zinc-containing toothpaste slurry three times a day for 14 days. The design of study was a crossover one. RESULTS: Microradiography employed in the in vitro study revealed the dentine surfaces treated with distilled water had a lower mineral content than those treated by zinc. Toothpaste with zinc had a statistically significant, 49% greater inhibitory efficacy on dentine demineralization over the control. In the in situ study, zinc-containing toothpaste was also confirmed to have an inhibitory effect on dentine demineralization when compared to the zinc-free toothpaste. These in vitro and in situ studies revealed that zinc oxide can have a significant effect on the inhibition of dentine demineralization, even when the zinc oxide is blended into toothpaste. SIGNIFICANCE: This suggests that zinc oxide may be effective in the prevention of root caries. PMID- 16046231 TI - Investigations of step-growth thiol-ene polymerizations for novel dental restoratives. AB - OBJECTIVES: The goal of this work was to investigate the feasibility of formulating novel dental restorative materials that utilize a step-growth thiol ene photopolymerization. Particularly, we are aiming to significantly reduce the polymerization shrinkage and shrinkage stress while retaining adequate physical properties as compared to current dimethacrylatre-based systems. METHODS: The thiol-ene system is composed of a 4:3 molar mixture of triallyl-1,3,5-triazine 2,4,6-trione (TATATO) and pentaerythritol tetramercaptopropionate (PETMP). The simultaneous measurement of shrinkage stress and functional group conversion was performed. Solvent extraction of unreacted monomers and dynamic mechanical analysis on the polymer networks that were formed were also studied. Flexural strength was measured for both filled and unfilled PETMP/TATATO and Bis GMA/TEGDMA systems. RESULTS: Photopolymerization of PETMP/TATATO occurs at a much higher rate, with the maximum polymerization rate six times faster, than Bis GMA/TEGDMA cured under the identical conditions. The results from the simultaneous measurement of shrinkage stress and conversion showed that the onset of shrinkage stress coincides with the delayed gel point conversion, which is predicted to be 41% for the 3:4 stoichiometric PETMP/TATATO resin composition. The maximum shrinkage stress developed for PETMP/TATATO was about 0.4 MPa, which was only approximately 14% of the maximum shrinkage stress of the Bis-GMA/TEGDMA system. Adequate flexural strength and flexural modulus values were obtained for both filled and unfilled PETMP/TATATO systems. SIGNIFICANCE: The dramatically reduced shrinkage stress, increased polymerization rate, significance increased functional group conversion, and decreased leachable species are all benefits for the use-of thiol-ene systems as potential dental restorative materials. PMID- 16046232 TI - Thiol-ene oligomers as dental restorative materials. AB - OBJECTIVE: The aim of this work was to prereact thiol-ene monomers to create reactive thiol or vinyl (ene)-functionalized oligomers, and to investigate the use of these materials as novel dental restorative material. Investigation has focused on the application of oligomeric thiol-ene materials as dental restorative resins with lower polymerization shrinkage and polymerization stress as compared to monomeric thiol-ene systems and particularly with respect to current dimethacrylate-based systems. METHODS: Reactive thiol-functionalized oligomers were created via photopolymerization using triallyl-1,3,5-triazine 2,4,6-trione (TATATO), trimethylolpropane tris(3-mercaptopropionate) (trithiol) and pentaerythritol tetramercaptopropionate (tetrathiol). Kinetic and mechanical investigation of Bis-GMA/TEGDMA, and oligomeric and monomeric thiol-ene systems were conducted. More specifically, polymerization shrinkage and stress, polymerization kinetics, glass transition temperature, flexural strength and flexural modulus were evaluated. RESULTS: Upon evaluation, the polymerization stress of oligomeric thiol-ene systems was dramatically reduced by as much as 33% when compared with the stress exhibited by monomeric thiol-ene systems and as much as a 92% reduction in stress relative to the current dimethacrylate-based dental restorative materials. Furthermore, the flexural strength and modulus of the monomeric and oligomeric thiol-ene resins were not significantly different. SIGNIFICANCE: Oligomeric thiol-ene systems offer potential as alternative dental restorative resins due to the significant reduction in polymerization shrinkage and stress while retaining the mechanical properties of monomer-based thiol-ene resins. PMID- 16046233 TI - In vitro and in vivo studies on blends of isotactic and atactic poly (3 hydroxybutyrate) for development of a dura substitute material. AB - Blends of semicrystalline isotactic poly(3-hydroxybutyrate) (PHB) with amorphous atactic PHB (at-PHB) were prepared by solution-casting using 30%, 50% and 70% at PHB, and were studied for medical applications. The mechanical properties of the blends including the elastic modulus and elongation at break are strongly affected by the blend composition. The elastic modulus decreases with increasing fraction of at-PHB in the blend from 3350 MPa in the case of PHB to 170 MPa of PHB/at-PHB (30/70). In contrast, the elongation at break increases from 2% in pure PHB up to 50% in the case of the blend with 70% at-PHB. The in vitro degradation is changed as well. The molecular weight of PHB/at-PHB (30/70) is reduced to 5% after 2 years storage in phosphate buffer compared to 35% for pure PHB stored at identical conditions. The in vitro cell vitality is slightly reduced depending on the composition. PHB/at-PHB blends with 30% and 50% at-PHB were selected as dura substitute in minipigs based on the results of the in vitro investigation and the mechanical testing. Patch films with a structured surface on one side were fabricated by a dipping-leaching method. Dura defects were clinically and histologically examined 3, 6, and 9 months after implantation, confirming defect closure, prevention of adhesions to brain tissue, and no signs of inflammation or malignant degeneration. The PHB-based patch materials fulfill the requirements which are necessary for a dura substitute. PMID- 16046234 TI - Progression of myelodysplasia to acute lymphoblastic leukaemia: implications for disease biology. AB - Myelodysplastic syndrome (MDS) comprises a group of clonal haematopoietic disorders characterized by peripheral blood cytopenias, bone marrow hypercellularity, and abnormal blood cell differentiation. Approximately 30% of cases of MDS eventually progress to acute myelogenous leukemia (AML), while progression of MDS into acute lymphoblastic leukemia (ALL) is rare. In this report, we describe a case of MDS that progressed to ALL, and review the 21 previously reported cases of MDS to ALL transformation. We review the cancer stem cell model and its application to these disorders, and discuss the implications of the rarity of transformation of MDS to ALL for the biology of MDS and the pathogenesis of ALL. PMID- 16046235 TI - Etodolac induces apoptosis and inhibits cell adhesion to bone marrow stromal cells in human myeloma cells. AB - OBJECTIVES: Cyclooxygenase-2 (COX-2) is reported to regulate apoptosis and to be an important cellular target for therapy. METHODS: We examined whether etodolac, meloxicam, and thalidomide inhibited proliferation and induced apoptosis in myeloma cell lines (RPMI 8226 and MC/CAR cells). RESULTS: Etodolac induced apoptosis more strongly compared with thalidomide or meloxicam. Etodolac induced down-regulation of Bcl-2 protein and mRNA, activation of Caspase-9, -7 and -3, cIAP-1 and Survivin, and loss of mitochondrial membrane potential in a dose dependent manner. In addition, when myeloma cells were coincubated with 50 microM etodolac on bone marrow stromal cells (BMSCs), myeloma cell adhesion to BMSCs was significantly inhibited compared with thalidomide or meloxicam coincubation, and the adhesion molecules VLA-4, LFA-1 (CD11a), CXCX4, and CD44 were suppressed on myeloma cells treated with etodolac. Moreover, 50-100 microM racemate of etodolac significantly inhibited the proliferation of myeloma cells compared to 100 microM R-etodolac or S-etodolac. CONCLUSIONS: Etodolac induced loss of mitochondrial membrane potential and apoptosis via a COX-2-independent pathway, suppressed the expression of adhesion molecules, and inhibited myeloma cell adhesion to BMSCs compared with thalidomide or meloxicam. The activities of etodolac potentially extend to the treatment of patients with myeloma resistant to standard chemotherapy, including thalidomide. PMID- 16046236 TI - Immunoglobulins and immunoglobulin genes of the horse. AB - Antibodies of the horse were studied intensively by many notable immunologists throughout the past century until the early 1970's. After a large gap of interest in horse immunology, additional basic studies on horse immunoglobulin genes performed during the past 10 years have resulted in new insights into the equine humoral immune system. These include the characterization of the immunoglobulin lambda and kappa light chain genes, the immunoglobulin heavy chain constant (IGHC) gene regions, and initial studies regarding the heavy chain variable genes. Horses express predominately lambda light chains and seem to have a relatively restricted germline repertoire of both lambda and kappa chain variable genes. The IGHC region contains eleven constant heavy chain genes, seven of which are gamma heavy chain genes. It is suggested that all seven genes encoding IgG isotypes are expressed and have distinct functions in equine immune responses. PMID- 16046237 TI - Nonylphenol-induced thymocyte apoptosis involved caspase-3 activation and mitochondrial depolarization. AB - Although the effect of 4-nonylphenol on cells of immune system have long been recognized, little is known about the effect of 4-nonylphenol on the induction of apoptosis and related signaling events in the lymphoid cells. In the present study, we used cultured thymocytes of mice to investigate the ability of 4 nonylphenol to induce the apoptosis of thymocytes and to explore the role of signal transduction pathway leading to apoptosis. The results showed that the cytotoxic effects of 4-nonyphenol involved DNA fragmentation (DNA ladder), characteristic of apoptosis. Staining of 4-nonyphenol-treated thymocytes with DNA binding fluorochrome Hoechst 33258 showed the typical apoptotic nuclei condensation and fragmentation of chromatin. The rates of apoptosis of the 4 nonylphenol-treated thymocytes increased significantly at 4 and 6 h, which were determined by analysis of hypodiploid cells and FITC-Annexin V and PI double staining. Flow cytometer analysis also revealed that the loss of mitochondrial membrane potential and increased activity of caspase-3 occurred concomitantly with the onset of 4-nonyphenol-induced apoptosis. Furthermore, a caspase-3 inhibitor, z-DEVD-fmk protected thymocytes from apoptosis induced by 4 nonyphenol. These results suggest that 4-nonylphenol induces thymocyte apoptosis via caspase-3 activation and mitochondrial depolarization. PMID- 16046238 TI - Hematopoietic prostaglandin D2 synthase in the chicken Harderian gland. AB - The Harderian gland (HG), a sero-mucous secreting organ in the eye orbit, has long been recognized as immunologically important in chickens. During experimentation to characterize immune components of the gland, proteomics analysis revealed the presence of hematopoietic prostaglandin D synthase (H PGDS). Extraction of total RNA followed by RT-PCR produced cDNA of 597 base pairs. DNA sequencing revealed nucleic acid and predicted amino acid sequences that were 99% aligned with the one published sequence for chicken H-PGDS of the spleen. Alignment with murine, rat, and human H-PGDS were 69, 69, and 66%, respectively. Ocular vaccination of chickens with a Newcastle Disease/Infectious Bronchitis vaccine (Mass.-Ark. Strain) induced an increase in H-PGDS expression determined by real-time PCR. Furthermore, immunohistochemistry of frozen HG sections showed positive stained cells for both H-PGDS and mast cell tryptase in the sub-epithelial cell layers of the HG ducts. Based on the potent vasoactive role of PGD(2), it appears that the chicken HG is a site of active mucosal immunity partially mediated by PGD(2) synthesized by H-PGDS in the gland. PMID- 16046239 TI - Fluctuation of the cytokine expression in the liver during the chronic woodchuck hepatitis virus (WHV) infection is not related to viral load. AB - The woodchuck together with the woodchuck hepatitis virus (WHV) is an excellent model to study the pathogenesis of hepadnaviral infections. Chronic WHV infection causes severe liver disease and hepatocellular carcinoma in woodchucks. The mechanism of viral clearance is not fully understood, interferons seem to play a major role in down-regulating viral replication prior to elimination of infected hepatocytes. We investigated on the pattern of cytokine and T-cell-marker expression in livers of woodchucks chronically infected with WHV. RNase protection-assay (RPA) was used to determine mRNA of woodchuck specific genes (TNF-alpha, IFN-gamma, IL-15, CD3, CD4, CD8). Serial liver biopsies were performed daily or weekly in eight chronic WHV-carrier woodchucks. Cytokine/T cell-marker expression differed significantly between the time points up to +/ 50% within each woodchuck. The different expression patterns of cytokines or T cell-markers did not correlate to the (weak) fluctuations in the viremia but may explain the observed fluctuations in the WHV/HBV-load in chronically infected individuals. Furthermore, we observed associations between cytokine and T-cell marker expression. The marginal fluctuations in viremia during the chronic infection may indicate, that, once the chronic hepadnaviral infection is established, cytokines/interferons expressed endogenously (i.e. not vector-borne or injected) play only a minor role. PMID- 16046240 TI - Antiviral drug discovery and development: where chemistry meets with biomedicine. AB - The successful development of antiviral drugs is highly dependent on a close interaction and collaboration between the chemist and the biologist (biomedic). This is illustrated by a number of representative examples: S adenosylhomocysteine (SAH) hydrolase inhibitors which display broad-spectrum antiviral activity, bromovinyldeoxyuridine (BVDU) and derivatives thereof, that are highly selective inhibitors of varicella-zoster virus (VZV), (dideoxy)nucleoside reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs) which are now widely used in the treatment of HIV infections (AIDS), the bicyclams (i.e. AMD3100) which were originally discovered as anti-HIV agents, then found to be potent CXCR4 antagonists and now being pursued for a number of indications such as stem cell mobilization, and the acyclic nucleoside phosphonates which have heralded a new strategy for the treatment of various DNA virus (herpes-, adeno-, pox-, papillomavirus) infections (cidofovir), hepatitis B (adefovir) and AIDS (tenofovir). PMID- 16046241 TI - The toxicokinetics of cadmium in carp under normoxic and hypoxic conditions. AB - Temporal depletion of oxygen, i.e. hypoxic events, frequently occurs in natural waters. It has been suggested that accumulation of micro-pollutants increases in aquatic animals as a result of an increased ventilation rate during such occasions. The observed increased toxicity under hypoxia of micro-pollutants may support this hypothesis, but for heavy metals the available uptake studies are contradictory. The present study tests whether accumulation of cadmium in common carp, Cyprinus carpio (L.) is increased under hypoxia and if the toxicokinetics are altered. A cadmium toxicity study was performed in which the cadmium uptake rates were determined using the radiotracer 109Cd under hypoxia and normoxia. The cadmium toxicokinetics were studied with radiotracer experiments at 100% air saturation, 50%, and 25% saturation from 6.5 nmol/L Cd at 25 degrees C. We could confirm the higher sensitivity of carp to cadmium under hypoxia. Hypoxic conditions did not influence the uptake rates or the accumulation dynamics. Therefore, the increased sensitivity of carp for Cd under hypoxia cannot be explained by a higher Cd body burden, initiated by a higher uptake rate or lower elimination rate under hypoxia. Additional, possible indirect effects, such as internal anoxia due to gill damage, could play a role in Cd toxicity under hypoxia. PMID- 16046242 TI - Prevalence and correlates of metabolic syndrome (MS) in older adults. AB - Metabolic syndrome (MS) is known to increase risks for cardiovascular disease. Risks for cardiovascular disease also increase with aging. The aim of the study was to describe prevalence and correlates of MS in older adults. Patients aged 65 years and over without any acute illness that were referred to our geriatrics unite for comprehensive geriatric assessment were included in this cross sectional study. MS was defined by using the WHO and the National Cholesterol Education Program (NCEP) definitions. The correlates were age, gender, low-grade inflammation as assessed by C-reactive protein (CRP) levels, high-homocysteine, total and LDL cholesterol, lipoprotein-a (Lip-a), apolipoprotein-A (Apo-A), apolipoprotein-B (Apo-B), nutrition point, coronary artery disease (CAD) and cerebrovascular event history. Total 1255 patients, 789 (62.9%) females, 466 (37.1%) males with a mean age of 71.8+/-6.3 years were included in our study. MS prevalence was 16.2% with WHO definition and 23.8% with NCEP definition. Prevalence of CAD in MS and non-metabolic syndrome (non-MS) patients was 38.4% versus 29.5% (p=0.010) in WHO and 35.3% versus 29.6% (p=0.066) in NCEP group, respectively. Prevalence of cerebrovascular event history was 11.3% versus 6.2% in WHO (p=0.008) and 9.9% versus 6.1% (p=0.026) in NCEP group, respectively. Multiple logistic regression analysis was performed to investigate the independent association of variables with the MS. Female sex, high-homocysteine, low-grade inflammation, CAD and cerebrovascular event history was found to be associated with both modified WHO and NCEP MS groups in the multivariate analysis. This study has shown that MS is common in elderly patients with strongly related to CAD and cerebrovascular events. Low grade inflammation as assessed by CRP and high-homocysteine level is strongly related to MS. PMID- 16046243 TI - Metalloproteinase activity is the sole factor responsible for the growth promoting effect of conditioned medium in Trichoplusia ni insect cell cultures. AB - Conditioned medium (CM) taken from a serum-free culture of Trichoplusia ni (BTI Tn-5B1-4, High Five) cells on days 2 and 3, shortened the lagphase and increased the maximum cell density when added to T. ni cultures with low-inoculum cell density. Gel filtration fractions of CM, eluting at around 45kDa, stimulated cell proliferation even better than CM. A protein in the gel filtration fraction was identified by N-terminal amino acid sequencing as a proteinase, related to a snake venom metalloproteinase. Casein zymography showed, multiple metalloproteinase bands between 48 and 25kDa, as well as precursor forms above 48kDa. Metalloproteinase bands below the main band at 48kDa were autocatalytic degradation products. Metalloproteinase activity was the sole factor responsible for the growth stimulating effect of CM as shown by using the specific metalloproteinase inhibitor dl-thiorphan. Metalloproteinases have recently been shown to release growth factors from sequestering extracellular proteins. We propose that the metalloproteinase is involved in autocrine regulation of T. ni proliferation in serum-free media. In addition, a gel filtration fraction of CM, eluting at about 10kDa, inhibited cell growth. Apart from a lysozyme precursor protein and a cyclophilin-like protein, a kazal-type proteinase inhibitor could be identified in this fraction. PMID- 16046244 TI - Nucleolar localization of non-structural protein 3b, a protein specifically encoded by the severe acute respiratory syndrome coronavirus. AB - The open reading frame 3 (ORF3) of the severe acute respiratory syndrome coronavirus (SARS-CoV) genome encodes a predicted 154-amino acid protein, which lacks similarities to any known protein, and is named 3b. In this study, it was shown that 3b protein was predominately localized to nucleus with EGFP tag at its N- or C-terminus. The localization patterns were similar in different transfected cells. Immuno-fluorescence assay revealed that 3b protein was co-localized well with C23 in nucleolus. C23, B23 and fibrillarin all are important nucleolar proteins, which localize in the region of the nucleolus. Co-transfection of p3b EGFP with pC23-DsRed, pB23-DsRed and pfibrillarin-DsRed further confirmed 3b's nucleolus localization. With construction of serial truncated mutants of 3b, a region (residues 134-154 aa) responsible for nucleolar localization was determinated in 3b protein. These results provide a new insight for further functional studies of SARS-CoV 3b protein. PMID- 16046245 TI - Motivating operations in appetite research. AB - Appetite research frequently employs principles derived from behaviour analysis. However, it has yet to utilise the more recent theoretical advances in this field. This paper describes the concept of the motivating operation (MO)--a behaviour analytic formulation of motivation. An MO is an environmental event that (a) establishes or abolishes the reinforcing or punishing effect of another event and (b) evokes or abates behaviours associated with that event. The paper describes both unconditioned and conditioned MOs and the ways in which they may help account for a variety of eating behaviours. It then goes on to highlight the main ways in which the MO account differs from other theories of motivation employed in appetite research. These relate to (1) the ways in which they account for non-regulatory feeding, (2) the extent to which they address cognitive variables and (3) their underlying philosophical assumptions and subsequent relation to intervention. PMID- 16046246 TI - Everyday dietary behaviour and the relationship between attention and meal size. AB - Previously, it has been demonstrated that overeating can occur when an individual engages in a cognitive task during a meal. One possibility, therefore, is that task performance can provide a measure of attention to dietary control. To explore this idea we measured performance on an otherwise irrelevant task during a meal. In two experiments, we compared intake and performance across five groups of females; dieters, and four groups of non-dieters, each with contrasting high and low scores on the restraint scale of the Dutch Eating Behaviour Questionnaire and the disinhibition scale of the Three-Factor Eating Questionnaire. Our analysis suggests that a correspondence can exist between task performance and intake (Experiment 1) and this might be influenced by the extent to which foods are regarded as forbidden (Experiment 2). However, aspects of our data are difficult to reconcile with the idea that task performance reflects a simple measure of attention to dietary control. Instead, one possibility is that individuals differ in the way that they choose to allocate attention during a meal. In relation to this idea, we discuss the possibility that some individuals choose to avoid food-related cognition by engaging strategically with other aspects of their cognitive environment. PMID- 16046247 TI - Feeding effects of melanocortin ligands--a historical perspective. AB - The process of energy homeostasis is a highly regulated process involving interacting signals between a variety of anorexigenic and orexigenic peptides, proteins and signaling molecules. The melanocortin system is an important component of this complex regulatory network. Involvement of the melanocortin pathway in the control of food intake and body weight regulation has been studied extensively in the past two decades. Previous studies that involve central administration of melanocortin molecules and examination of molecules that effect food intake in melanocortin knockout (KO) mice (MC3R, MC4R, POMC, AGRP and NPY) have been examined. In this review, we have summarized feeding studies that have resulted in the recognition of the melanocortin system as a major contributor to the complex neuroendocrine system regulating energy homeostasis. PMID- 16046248 TI - How to build an "active" patient? The work of AIDS associations in France. AB - "What is an "active" patient?" is a question that arises in most medicine and illness-related social science research. This article examines the normative work carried out by AIDS associations in France to define an "active" patient in healthcare and research. While the fight against AIDS is often presented as being homogenous, we look at the diversity of opinion between different associations (Aides, Act Up-Paris, Actions Traitements and Positifs). We find four different cases: the patient as manager of his illness, the empowerment of patients, the science-wise patient and the experimenter. Systematic comparison of these cases shows that these perceptions of the "active" patient, in terms of the same pathology, are based upon different ways of seeing: the nature of the relationships between the different types of knowledge of the illness (scientific knowledge, clinical knowledge, experience of the illness) and the distribution of roles and powers among the various actors in the healthcare system (the government, pharmaceutical companies, the medical profession, the patients). This article highlights the historical dynamics which allow us to have a better understanding of these differences, especially the major distinction between two generations of associations, which adopted different positions with regard to their public identity. PMID- 16046249 TI - Aggregation of fuzzy views of a large number of stakeholders for multi-objective flood management decision-making. AB - Flood management decision-making problems are often associated with multiple objectives and multiple stakeholders. The challenge remains on how to obtain the diversified opinions of a large number of stakeholders where uncertainty plays a major role. In response to this challenge, a methodology has been proposed to capture the views of multiple stakeholders using fuzzy set theory and fuzzy logic. Three possible forms of individual stakeholders' input, scale (crisp), linguistic (fuzzy), and conditional (fuzzy), are analyzed to obtain the aggregated input by using Fuzzy Expected Value. The methodology is applied to flood management in the Red River Basin, Manitoba, Canada. The outputs derived from the three different input types show good correlation. Thus, the proposed methodology shows alternative ways of soliciting the opinion of a large number of stakeholders and aggregating those opinions for use in multi-objective decision making. PMID- 16046250 TI - Removal of Pb(II) from wastewater using wheat bran. AB - The adsorption of Pb(II) ions from aqueous solutions on wheat bran (WB) has been investigated as a function of initial concentration, adsorbent dose, adsorbent particle size, agitation speed, temperature, contact time and pH of solution. The equilibrium process was described well by the Langmuir isotherm model with maximum sorption capacities of 69.0, 80.7 and 87.0 mgg(-1) of Pb(II) on wheat bran at 20, 40 and 60 degrees C, respectively. Thermodynamic parameters, i.e. DeltaG(0), DeltaH(0) and DeltaS(0) have also been calculated for the system and the sorption process was found to be endothermic. Good correlation coefficients were obtained for the pseudo second-order kinetic model. The metal ion could be stripped by addition of 0.5M HCl, making the adsorbent regeneration and its reutilization possible. PMID- 16046251 TI - Multi-criteria natural resource management with preferentially dependent decision criteria. AB - The traditional assumption with multi-criteria natural resource management models is that the values of the decision alternatives with respect to one criterion can be assessed independently of the values of the decision alternatives with respect to other criteria. In practice, however, the assumption of independent decision criteria is not always realistic. For example, the importance of the amount of old forests to biodiversity may depend on the amount of dead wood. This paper shows how multi-criteria natural resource management problems can be analysed in a case involving dependent decision criteria. The proposed models are based on pair-wise comparisons of items in the ratio scale and statistical regression analysis of the pair-wise comparisons data. The basic idea behind the dependent models is to ask the decision maker (e.g. forest owner) to assess the decision alternatives or their characteristics simultaneously with respect to dependent decision criteria. The separate models for discrete and continuous cases are given, which enable the number of potential management alternatives to vary from a few to infinite. The models proposed allow a more realistic description of the decision problem in multi-criteria natural resource management. PMID- 16046252 TI - Functions of CXCL12 and CXCR4 in breast cancer. AB - The chemokine CXCL12 (SDF-1) and its cognate receptor CXCR4 were first identified in the context of trafficking and homeostasis of immune cells, such as T lymphocytes. Subsequently, it has been determined that CXCR4 regulates several key processes in a wide variety of cancers. Functions of CXCL12 and CXCR4 in cancer first were described in metastatic breast cancer, and more recent studies also have identified roles for this signaling pathway in primary breast tumors. This review focuses on functions of CXCR4 and CXCL12 in primary and metastatic breast cancer, including molecular mechanisms of action and relationships of this pathway to other key regulators of breast cancer progression. We also describe pre-clinical studies indicating the potential to exploit CXCR4 as a new molecular target for diagnosis and treatment of breast cancer in patients. PMID- 16046253 TI - Lifetime and regeneration of immobilized titania for photocatalytic removal of aqueous hexavalent chromium. AB - Immobilized titania (TiO2) batch reactors reduced hexavalent chromium (Cr(VI)) in the form of potassium dichromate (K2Cr2O7) to trivalent chromium (Cr(III)) in aqueous solution at pH 3 under 171 W/m2 light intensity. The light source was a 125-W ultraviolet (UV) lamp. The Cr(VI) reduction showed zero-order kinetics (k0), while the Cr(VI) adsorption fitted with first-order kinetics (k(1st)). Adsorption capacity increased with increasing initial Cr(VI) concentration, and the area of immobilized TiO2 limited the reduction efficiency. The lifetime of fresh immobilized TiO2 was approximately 14 h. In addition, the regeneration of TiO2 with 3M sodium hydroxide (NaOH) was necessary to improve adsorption reaction. PMID- 16046254 TI - In vitro penetration of fresh and vitrified swine oocytes by homologous spermatozoa using different incubation systems. AB - The present study consisted of two experiments. In the first one, ejaculates from four boars were used to compare in vitro penetration (IVP) rates of fresh and vitrified swine oocytes by homologous spermatozoa in four treatments: fresh oocytes in conventional incubation (CO2 incubator) (FC), vitrified oocytes in conventional incubation (VC), fresh oocytes in submarine (bag) incubation (FS) and vitrified oocytes in submarine incubation (VS). The IVP rates for FC, VC, FS and VS were 46.5, 44.3, 36.9 and 33.1%, respectively. Analysis through Chi-square tests identified no differences in IVP rates between FC and VC and between FS and VS (P > 0.05), but IVP rate for FC was greater (P < 0.05) than those for both FS and VS. Besides IVP rate for VC did not differ (P > 0.05) from those for FC and FS, but it was greater than that for VS (P < 0.05). Logistic regression analysis identified differential effects of treatments dependant on individual boars. The second experiment evaluated the influence of semen storage period on the semen quality of the two boars associated with greater IVP rate in the first experiment. Semen quality was estimated by IVP rate using the VC treatment and by the following methods: sperm motility, sperm morphology, hypoosmotic swelling test (HOST) and thermal stress test (TST). According to analysis using Chi-square tests, IVP rate did not differ (P > 0.05), for the first boar, between 0 (100.0%) and 24 h of semen storage (98.1%) nor after 48 and 72 h (66.0 and 59.3%, respectively), but IVP rates were greater during the 0-24 h period compared with the 48-72 h period (P < 0.05). For the second boar, IVP rate at 0 h (50.6%) was greater (P < 0.05) than at 24, 48 and 72 h of semen storage (34.3, 28.3 and 24.0%, respectively), with no further differences observed after 24 h (P > 0.05). Logistic regression analysis identified that the effect of storage on IVP rate was influenced by the effect of individual boars. No differences in semen quality during the storage period were identified by conventional methods of semen evaluation, for either boar (P > 0.05) using analysis of variance with repeated measures. These results indicate that IVP test can be used to estimate boar fertility, even when vitrified oocytes are used (if using conventional CO2 incubators) or using an alternative submarine incubation system (if using fresh oocytes). The IVP test was the only method of semen evaluation that identified the reduction in semen quality up to 72 h of storage. PMID- 16046255 TI - Endocrine profiles and reproductive output in European ground squirrels after unilateral ovariectomy. AB - In the course of a long-term study on reproductive physiology and behaviour in female European ground squirrels (Spermophilus citellus) we investigated interactions among reproductive timing, maternal effort and hibernation. To gain information on the state of follicular development shortly before hibernation, histological investigations of ovaries were carried out. To collect the ovaries, unilateral ovariectomy was chosen as the least invasive method. The aim of this study was to monitor patterns of hormone secretion, behavioural parameters and reproductive output in females after unilateral ovariectomy, and compare them with intact individuals to assess potential effects of the treatment. Reproductive performance including duration of gestation and lactation, litter size, and offspring mass at natal emergence were similar in treated and untreated individuals. Before weaning, plasma estradiol levels were higher in the hemicastrated group than in intact females. Progesterone concentrations tended to be lower in unilaterally ovariectomized females during gestation and postlactation. In both groups we measured cortisol metabolites in fecal samples to estimate basal adrenal activity. Untreated individuals had significantly higher cortisol levels than hemicastrated females during lactation. In all other phases no significant differences in patterns of cortisol excretion were found between the two groups. Prehibernatory body-mass did not differ between treated and untreated females. These results indicate that unilateral ovariectomy had no significant negative effects on reproductive timing and output in female European ground squirrels. Compensatory activity of the remaining ovary does occur, concerning estradiol secretion even a kind of overcompensation was reported. These elevated estradiol levels in unilaterally ovariectomized females could play a role in limiting the degree of follicular compensation. PMID- 16046256 TI - Improving the therapeutic efficacy of peptides and proteins: a role for polysialic acids. AB - Peptide and protein drugs are a growing class of therapeutics. However, their effective application in the clinic is compromised by problems, for instance proteolysis in the circulating blood, premature clearance through the kidneys, and immunogenicity. A number of approaches have been used to circumvent such shortcomings including changes in the primary peptide structure, entrapment into nanoparticles (e.g. liposomes) and conjugation to polymers. Polysialylation, namely, conjugation of peptides and proteins to the naturally occurring, biodegradable alpha-(2-->8) linked polysialic acid is a recent development, which promises to be at least as effective as PEGylation but without its potential toxicity. Polysialylation of a range of peptide and protein therapeutics has led to markedly reduced proteolysis, retention of their activity in vivo, prolongation of their half-life in the circulation and reduction in immunogenicity and antigenicity. It is anticipated that polysialylation will lead to a new generation of peptide and protein constructs with significantly improved pharmacological profiles. PMID- 16046257 TI - Reliability and validity of the Child Behavior Checklist Obsessive-Compulsive Scale. AB - This study examined the psychometric properties of the Obsessive-Compulsive Scale (OCS) of the Child Behavior Checklist (CBCL). Participants included 48 youth with obsessive-compulsive disorder (OCD), 41 with a non-OCD internalizing disorder, and 101 with an externalizing disorder. Confirmatory factor analysis of the 8 item OCS did not result in an adequate fit. Exploratory factor analysis identified a 1-factor model consisting of 6 items. Adequate internal consistency for the revised OCS (OCS-R) was obtained, and convergent validity was supported by moderate relationships with other OCD indices. The OCS-R had stronger associations with measures of OCD symptoms than with measures of depression and externalizing behaviors. Youth with OCD had significantly higher OCS-R scores than those with internalizing and externalizing disorders. Suggestions for cutoff scores are provided using results from ROC analyses. Overall, these findings suggest that the OCS-R is a reliable and valid instrument for the assessment of pediatric OCD. PMID- 16046258 TI - Association between RANTES promoter polymorphism -401A and enhanced RANTES production in atopic dermatitis patients. PMID- 16046259 TI - Childhood respiratory infections and hospital admissions for COPD. AB - BACKGROUND: Chronic Obstructive Pulmonary Disease (COPD) exacerbations are associated with viral infections. We wished to determine if respiratory viral infection of children in the community was associated with hospital admissions of patients with exacerbations of COPD. METHODS: We collected data over a 45-month period from the Northern Ireland Regional Virus Laboratory and from a general hospital in the same locality. We studied the relationship between upper respiratory infections in children and COPD admissions. We also examined the role of school holidays. RESULTS: Correlations were seen between the frequency of all viral infections in children and the number of adult COPD hospitalizations (P<0.005). Subgroup analysis showed distinct relationships with epidemics of; influenza A (P<0.001), influenza B (P<0.05), adenovirus (P=0.05), respiratory syncytial virus (P<0.005) and hospital admissions of patients with COPD. There were significantly fewer COPD admissions in the week after the start of a school holiday period (P<0.05). CONCLUSIONS: When children are hospitalized with viral respiratory infection there is an associated rise in adult COPD admissions. This suggests exacerbations of COPD are associated with epidemics of respiratory viruses. When children are on school holidays there is a reduction in COPD admissions in the community. This provides further support for respiratory viruses in the pathogenesis of COPD exacerbations. PMID- 16046260 TI - Do current treatment protocols adequately prevent airway remodeling in children with mild intermittent asthma? AB - BACKGROUND: Asthma treatment per Global Initiative for Asthma (GINA) guidelines targets airway remodeling and achievement of normal lung function. OBJECTIVE: To study inhaled steroid therapy on airway remodeling and achievement of normal lung function. SUBJECTS AND SET UP: Asthmatic patients were followed over 5 years . The children were divided into two groups. Group I (148 children-treated group) consisted of all the children using inhaled steroids on a regular basis and group II (50 children-untreated group) included patients with mild intermittent asthma who did not use inhaled steroids and were treated with bronchodilators as needed. Detailed medication and compliance history were recorded and the children underwent regular pre- and post-bronchodilator spirometry and physician examination. RESULTS: The incidence of airways remodeling as defined by the low FEV1/FVC post-bronchodilator was markedly increased over the 5-year period in the untreated group compared to the treated group. In the treated group the FEV1/FVC ratio post-bronchodilator decreased from 35% to 20.9% (P<0.009%) compared with the untreated group where the number of children with low FEV(1)/FVC post bronchodilator increased from 10% to 28% by the end of the study period. CONCLUSION: Patients with mild asthma who do not receive regular inhaled steroids are at increased risk for airway remodeling. Therefore, we recommend that children with mild intermittent asthma undergo regular pulmonary function testing and that if any deterioration is detected in the FEV1/FVC ratio post bronchodilators; preventative inhaled corticosteroid therapy be initiated. PMID- 16046261 TI - Dietary recommendations to meet both endocrine and autocrine needs of Vitamin D. AB - In the most recent revision of the dietary recommendations for Americans and Canadians in 1997, a recommended intake for Vitamin D was set in the absence of an estimation of mean requirements. There are now new data to estimate average requirements; however, there must be consideration of factors affecting need in populations and of total body tissue needs including the prevention and treatment of cancer. A recent study provides dietary dose-response data in the absence of sun exposure, and a mean requirement of 12.5microg (500IU) was found for Caucasian men. A seasonal build up (summer) and waning (winter) of Vitamin D stores implies that the requirement of Vitamin D in complete absence of yearly summertime sun exposure would approach levels of intake that mimic Vitamin D gained from sun exposure. High prevalence of Vitamin D insufficiency and the re emergence of rickets have been observed worldwide. For many countries without mandatory staple food fortification, Vitamin D intake is often too low to sustain healthy circulating levels of 25 hydroxyvitamin D. Even in some countries that require (mandatory) or allow fortification (optional), Vitamin D intakes are low in some groups due to their unique dietary patterns, such as low milk consumption, vegetarian diet, limited or no use of dietary supplements, or changes away from traditional food consumption. Supplement use can significantly increase Vitamin D intakes across all age and gender groups but the benefit is primarily gained in persons whose intakes are close to adequate. African American men and women have greater prevalence of Vitamin D insufficiency, which may be a factor in their susceptibility to certain cancers. New recommendations for Vitamin D should be made for the otherwise healthy populations in greatest need of dietary Vitamin D due to lack of adequate sun exposure. PMID- 16046262 TI - The rationale for deltanoids in therapy for myeloid leukemia: role of KSR-MAPK C/EBP pathway. AB - The evidence for the promising potential for derivatives of Vitamin D (deltanoids) in the treatment of myeloid leukemias is increasing, but currently is not matched by the understanding of the precise mechanisms by which these anti neoplastic effects are achieved. Unlike solid tumors in which growth retardation by deltanoids appears to result from inhibition of cell proliferation and the promotion of cell death by apoptosis, control of myeloid leukemia proliferation by deltanoids results from the induction of differentiation of the immature myelo monocytic cells towards functional monocytic cells. We present here the accumulating evidence that a pathway that is initiated by deltanoid activation of Vitamin D receptor (VDR) and leads to monocytic differentiation of human myeloblastic HL60 cells, includes the MEK-ERK and JNK mitogen-activated protein kinases (MAPKs), their positive and negative regulators and a downstream effector C/EBPbeta. As in other cells, the abundance of VDR protein increases shortly after an exposure of HL60 cells to 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2) D(3)). Other early events include a parallel upregulation of kinase suppressor of Ras (KSR-1) and the activation of the ERK MAPK pathway and data suggest that KSR-1 acts to amplify the signal provided by low concentrations of 1alpha,25(OH)(2) D(3). Maintenance of monocytic differentiation may be enhanced by JNK, but diminished by p38, MAPK signaling. Downstream, one of the targets of these pathways is C/EBPbeta, which can directly interact with the promoter for CD14, a gene characteristically expressed in monocytes. Importantly, in freshly obtained acute myeloid leukemia (AML)-M2 cells exposed to PRI-2191, a novel deltanoid with a modified side chain, upregulation of C/EBPbeta paralleled the induction of monocytic differentiation. These data provide a basis for the hypothesis that deltanoid-induced upregulation of C/EBPbeta bypasses the block to granulocytic differentiation in myeloid leukemia cells by redirecting the cells to monocytic differentiation. PMID- 16046263 TI - Time-dependent 241Am activity in the environment from decay of 241Pu released in the Chernobyl accident. AB - The time-dependent activity of the alpha-emitting radionuclide 241Am, resulting from beta-decay of 241Pu present in the accidental release from Unit 4 of the Chernobyl NPP, has been studied. The ratios of the activity of 241Pu to those of 239, 240Pu and of 238, 239, 240Pu at the time of the accident have been derived as 69.6+0.6 and 47.1+0.4, respectively. Based on these results, a prediction of the future activity ratio 241Am/238,239,240Pu has been made, showing that it will increase slowly to the value 1 in the year 2005 and to a maximum value of 1.7 at the end of this century, followed by a gradual decrease. PMID- 16046264 TI - Tension pneumomediastinum after severe vomiting in a 21-year-old female. AB - A 21-year-old female with chronic membranoproliferative nephritis was admitted for suspected esophageal disruption and asthma after severe, prolonged vomiting. At the time of admission she presented with dyspnea, tachypnea, arterial hypotension and tachycardia. Physical examination showed discrete signs of ectopic air at the neck and distended cervical veins. CT-scan of the chest showed severe mediastinal emphysema with compression of the right atrium. After cervical mediastinotomy the cardiorespiratory parameters normalized immediately. Esophagoscopy showed multiple longitudinal mucosal tears between 25 and 45 cm; fluoroscopically, there was no leakage of contrast medium. Following conservative treatment the patient recovered completely and was discharged on day 8. PMID- 16046266 TI - Early open thoracotomy and mediastinopleural irrigation for severe descending necrotizing mediastinitis. AB - OBJECTIVE: Descending necrotizing mediastinitis (DNM) is a severe infection spreading from the cervical region to the mediastinal connective tissue. The mortality rate was reported as 40% until the 1980s. Since DNM is uncommon, few reports of large series of patients with DNM (i.e. more than 10 cases) have been published. The present aim was to evaluate our treatment strategy for DNM by retrospective chart review. METHODS: Retrospective chart review was performed in 10 patients with DNM between 1991 and 2003. The mean age was 53.8+/-23.3 years (median 58, range 16-82). The causes of DNM were primary peritonsillar or parapharyngeal abscess in 5 patients, post-extraction odontogenic abscess in 3, cervical abscess of post-tracheostomy in 1, and unknown in 1 patient. In nine cases, the abscess extended from the cervical region to the lower mediastinum. Immediately after the diagnosis of DNM, broad-spectrum antibiotics were administered empirically, and surgical treatments consisting of cervical drainage, thoracotomy with radical surgical debridement of the mediastinum and excision of necrotic tissue, decortication, and irrigation were performed in all cases. Post-operatively, mediastinopleural irrigation with saline was performed once or twice a day until a culture of pleural effusion became negative. RESULTS: The mean duration of chest tube retention was 26.7+/-17.0 days, and the mean hospital stay was 62.3+/-33.9 days. Five patients suffered from severe complications including septic shock, acute respiratory distress syndrome, disseminated intravascular coagulation, and pan-peritonitis due to duodenal perforation. The outcome was favorable in 8 patients. Of those with severe complications, two patients, who were older than 75 and had diabetes, died of multiple organ failure due to septic shock. Therefore, the mortality rate was 20%. CONCLUSION: Our treatment strategy for severe DNM was efficacious for early treatment and reduced the mortality rate. Early detection of DNM, and immediate thoracotomy and irrigation of the mediastinum and thoracic cavity, are recommended. PMID- 16046267 TI - Hybrid treatment of a symptomatic aortic and left subclavian false aneurysm. PMID- 16046269 TI - Genetic engineering of Escherichia coli for the production of NI,NII diacetylchitobiose (chitinbiose) and its utilization as a primer for the synthesis of complex carbohydrates. AB - Chitinbiose was produced at more than 4 g L-1 by a high cell density culture of an Escherichia coli strain that co-expressed the rhizobial chitinoligosaccharide synthase gene nodC and a truncated form of the chitinase gene chiA which has been designed to be functionally produced in the E. coli cytoplasm. Chitinpentaose, which has previously been shown to be produced by the nodC protein in growing E. coli, was formed as an intermediate that was subsequently hydrolyzed into chitinbiose by the chitinase encoded by chiA. Chitinbiose was mainly recovered in the extracellular medium and to prevent its catabolism, the genes for the chitinbiose PTS permease had to be disrupted. When the additional gene lgtB for beta1,4-galactosyltransferase was expressed, intracellular chitinbiose was converted into the trisaccharide Galbeta-4GlcNAcbeta-4GlcNAc which could serve as acceptor for glycosyltransferase that recognize the terminal N-acetyllactosamine structure. PMID- 16046270 TI - A case report of acute hepatitis due to brucellosis. PMID- 16046271 TI - Could iodine be effective in the treatment of human immunodeficiency virus and AIDS-associated opportunistic infections? PMID- 16046272 TI - Genetic diversity at 15 microsatellite loci among the Adi Pasi population of Adi tribal cluster in Arunachal Pradesh, India. AB - Genetic polymorphisms at 15 tetrameric short tandem repeat (STR) loci were studied in 203 healthy individuals of Adi Pasi population from Arunachal Pradesh, India. All the loci analyzed were highly polymorphic and there was no significant deviation from the Hardy-Weinberg equilibrium (HWE) excepting D8S1179 and D18S51. Other forensic useful statistical parameters were also calculated and the 15 microsatellite markers selected for this study were found to be suitable for human identification and population genetic studies. PMID- 16046273 TI - A fatal clomipramine intoxication case of a chronic alcoholic patient: application of postmortem hair analysis method of clomipramine and ethyl glucuronide using LC/APCI/MS. AB - Toxicological investigations of postmortem specimens of a 26-year-old man were performed with the use of LC/APCI/MS. They revealed in the blood of the deceased clomipramine (9.49 microg/g) and its main metabolite norclomipramine (1.10 microg/g) at concentrations explaining the fatal outcome. The presence of these xenobiotics in a 12-cm-long strand of hair (clomipramine, 7.60 ng/mg in I segment; 4.19 ng/mg in II segment; 1.86 ng/mg in III segment; norclomipramine, 5.71 ng/mg in I segment; 9.71 ng/mg in II segment; 4.13 ng/mg in III segment) confirmed the fact obtained from the medical history that the deceased had been receiving clomipramine as an antidepressant for 1 year prior to his death. The analysis demonstrated ethanol in autopsy blood (2.5mg/ml) and urine (3.2mg/ml); ethyl glucuronide as a marker of chronic alcohol abuse was detected in the deceased's hair (0.44 ng/mg in I segment; 0.07 ng/mg in II segment; n.d. in III segment). These findings may suggest the contribution of alcohol in the mechanism of drug-ethanol interaction, which in consequence might have affected the biotransformation of clomipramine in the final period of his life and evoked the ultimate toxic effect. PMID- 16046274 TI - Population data for 15 loci (Identifiler Kit) in a sample from the Valley of Mexico. AB - We analyzed 242 individuals from the Valley of Mexico, including the larger and more cosmopolitan city of this country. They were PCR-typed for 15 STR loci with the AmpFlSTR Identifiler PCR Amplification Kit (Applied Biosystems). Allele frequencies for each STR were estimated and compared to previous reports. Genotype distribution by locus and by two-loci combination was in agreement with Hardy-Weinberg expectations for all fifteen STRs. This STR system in Mexican mestizos presented a combined probability of exclusion (PE) and discrimination (PD) longer than 99.999%, respectively. PMID- 16046275 TI - Re-engineering the process of medical imaging physics and technology education and training. AB - The extensive availability of digital technology provides an opportunity for enhancing both the effectiveness and efficiency of virtually all functions in the process of medical imaging physics and technology education and training. This includes degree granting academic programs within institutions and a wide spectrum of continuing education lifelong learning activities. Full achievement of the advantages of technology-enhanced education (e-learning, etc.) requires an analysis of specific educational activities with respect to desired outcomes and learning objectives. This is followed by the development of strategies and resources that are based on established educational principles. The impact of contemporary technology comes from its ability to place learners into enriched learning environments. The full advantage of a re-engineered and implemented educational process involves changing attitudes and functions of learning facilitators (teachers) and resource allocation and sharing both within and among institutions. PMID- 16046276 TI - Nursing informatics knowledge and competencies: a national survey of nursing education programs in the United States. AB - An online survey of deans/directors of 266 baccalaureate and higher nursing programs in the U.S. was developed by informatics expert nurses. Participants (1) identified nursing informatics (NI) competencies and knowledge of undergraduate and/or graduate students in their nursing programs; (2) determined faculty preparedness to teach NI and to use informatics tools; and (3) provided perceptions of NI requirements of local practicing nurses. Frequency data and qualitative responses were analyzed. Approximately half of undergraduate nursing programs were teaching information literacy skills and required students to enter with word-processing and email skills. Least visible informatics content at all levels included the use of information system data standards, the Nursing Information and Data Set Evaluation Center criteria, the unified medical language system (UMLS), and the nurse's role in the life cycle of an information system. Almost 50% of respondents perceived faculty as "novice" and "advanced beginners" in teaching and using NI applications. Participants reported no future plans to offer NI training in their region. Findings have major implications for nurse faculty, staff developers, and program administrators who are planning continuing education opportunities and designing nursing curricula that prepare nurses for use of the electronic health record and 21st century professional practice. PMID- 16046277 TI - Focal cooling for epilepsy: an alternative therapy that might actually work. AB - The therapy of focal epilepsy remains inadequate. Many patients who have localization-related seizures find themselves either overmedicated with anticonvulsants or suffering from frequent seizures. While surgical resection can lead to excellent outcomes in up to 60% of patients with neocortical epilepsy, there are obviously many who either fail surgery or are deemed inappropriate surgical candidates. We are currently determining the efficacy of local cooling for the therapy of certain focal epilepsies. We have attempted to adapt new technologies borrowed from electrical and mechanical engineering to develop cooling devices that will ultimately improve the diagnosis and therapy of these focal epilepsies. The present review describes the rationale for this research and our progress to date. PMID- 16046278 TI - Episodic depersonalization in focal epilepsy. AB - In this report a patient with episodic depersonalization is described. As the depersonalization episodes had been attributed to partial seizures, this patient was treated with antiepileptic medication. However, clinical evaluation with long term video/EEG revealed no evidence of seizure activity during the depersonalization episodes. On the other hand, further evaluation revealed findings that are frequently associated with focal epilepsy. In addition to episodic depersonalization, this patient had secondary generalized seizures. The relationship between episodic depersonalization, temporal lobe pathology, and epilepsy is discussed against the background of this case. PMID- 16046279 TI - Consciousness, epilepsy, and emotional qualia. AB - The last decade has seen a renaissance of consciousness studies, witnessed by the growing number of scientific investigations on this topic. The concept of consciousness is central in epileptology, despite the methodological difficulties concerning its application to the multifaced ictal phenomenology. The authors provide an up-to-date review of the neurological literature on the relationship between epilepsy and consciousness and propose a bidimensional model (level vs contents of consciousness) for the description of seizure-induced alterations of conscious states, according to the findings of recent neuroimaging studies. The neurophysiological correlates of ictal loss and impairment of consciousness are also reviewed. Special attention is paid to the subjective experiential states associated with medial temporal lobe epilepsy. Such ictal phenomenal experiences are suggested as a paradigm for a neuroscientific approach to the apparently elusive philosophical concept of qualia. Epilepsy is confirmed to represent a privileged window over basic neurobiological mechanisms of consciousness. PMID- 16046280 TI - Does statin monotherapy address the multiple lipid abnormalities in type 2 diabetes? AB - Lipid abnormalities, which are common in type 2 diabetes, predispose to a greatly increased risk of coronary heart disease. This characteristic dyslipidaemia includes decreased concentrations of high-density lipoprotein cholesterol (HDL C), elevated triglycerides, and a small, dense, atherogenic form of low-density lipoprotein cholesterol (LDL-C). Insulin resistance and obesity, which is commonly present in type 2 diabetes, act in concert to disrupt normal lipoprotein metabolism; reverse cholesterol transport in particular. The proatherogenic changes, which result from this process include enrichment of very-low-density lipoprotein with cholesteryl esters and enrichment of LDL with triglycerides. Results from both the Pravastatin Pooling Project and the Heart Protection Study demonstrate that, although people with diabetes obtain the same relative risk reduction with statin therapy, the absolute benefit derived is much lower than for comparable individuals without diabetes. In order to achieve improved outcomes in diabetes patients, it will be important to address other abnormalities in their lipid profiles, including elevated triglycerides and low HDL-C. PMID- 16046281 TI - What is the consequence of an abnormal lipid profile in patients with type 2 diabetes or the metabolic syndrome? AB - Patients with type 2 diabetes have an atherogenic lipid profile, which greatly increases their risk of coronary heart disease (CHD) compared with people without diabetes. The largest disparity in lipid levels among people with and without diabetes occurs for high-density lipoprotein cholesterol (HDL-C) and triglycerides: triglycerides tend to be markedly higher and HDL-C moderately lower in patients with diabetes, in contrast to the negligible difference observed in low-density lipoprotein cholesterol (LDL-C) and total cholesterol. However, patients with type 2 diabetes are more likely to have the atherogenic form of LDL-C than people without diabetes, as well as low HDL-C, which restricts reverse cholesterol transport and may also be associated with increased lipid oxidation. Among patients who have suffered a myocardial infarction, increased LDL-C is apparent in early adulthood, whereas a detectable difference in HDL-C levels becomes increasingly apparent with age and most pronounced after age 60 years, compared with healthy controls. Evidence indicates that the increased risk of macrovascular complications of type 2 diabetes begins long before the onset of clinical hyperglycaemia. Despite successful reduction of LDL-C with statin therapy, patients continue to be at increased risk for CHD if their HDL-C levels remain suboptimal, in part due to persistence of enhanced lipid exchange. Observational data suggest that increasing HDL-C should be much more potent therapeutically than a similar proportionate decrease in LDL-C. PMID- 16046282 TI - Why dietary restriction substantially increases longevity in animal models but won't in humans. AB - Caloric restriction (CR) extends maximum longevity and slows aging in mice, rats, and numerous non-mammalian taxa. The apparent generality of the longevity increasing effects of CR has prompted speculation that similar results could be obtained in humans. Longevity, however, is not a trait that exists in a vacuum; it evolves as part of a life history and the physiological mechanisms that determine longevity are undoubtedly complex. Longevity is intertwined with reproduction and there is a cost to reproduction. The impact of this cost on longevity can be age-independent or age-dependent. Given the complexity of the physiology underlying reproductive costs and other mechanisms affecting life history, it is difficult to construct a simple model for the relationship between the particulars of the physiology involved and patterns of mortality. Consequently, we develop a hypothesis-neutral model describing the relationship between diet and longevity. Applying this general model to the special case of human longevity and diet indicates that the benefits of caloric restriction in humans would be quantitatively small. PMID- 16046283 TI - The neuroprotective actions of corticotropin releasing hormone. AB - Corticotropin-releasing hormone (CRH) modulates the activity of the hypothalamic pituitary-adrenal (HPA) axis, and has a key role in mediating neuroendocrine effects that occur in response to stressful stimuli. Disruption of the CRH system however has been shown to be closely associated with the progression of Alzheimer's disease (AD), and these observations prompted an investigation into the potential neuroprotective effects of the hormone. In addition to its regulatory affects on the molecular processes that underlie AD i.e., amyloid precursor protein (APP) processing and potentially tau phosphorylation, evidence is provided that the neuroprotective effects of CRH are mediated by a number of diverse mechanisms. These stem from activation of its high affinity receptor, the CRH type 1 receptor, and involve the induction of protective intracellular pathways including PKA-CREB that eventually lead to expression of neurotrophic factors. Conversely, inhibition of harmful events, such as caspase activation during apoptosis may also occur. Taken together, an impressive amount of evidence has accumulated recently, highlighting this new and potentially important function of CRH. PMID- 16046284 TI - Chromatin and the DNA damage response. AB - The impact of chromatin structure upon the DNA damage response is becoming increasingly apparent. We can reasonably expect many more papers showing how chromatin and chromatin modifications impact upon aspects of the DNA damage response. Here, we present our perspective on some recent developments in this exciting area of cell biology. We aim that this review will be of interest to those who study the DNA damage response, but not usually in the context of chromatin, and equally to those who study chromatin, but not the DNA damage response. It seems likely that these two communities will increasingly share common questions and interests. PMID- 16046285 TI - A new and simple solid-phase extraction method for LC determination of pyronaridine in human plasma. AB - A new approach using a simple solid-phase extraction technique has been developed for the determination of pyronaridine (PND), an antimalarial drug, in human plasma. After extraction with C18 solid-phase sorbent, PND was analyzed using a reverse phase chromatographic method with fluorescence detection (at lambda(ex)=267 nm and lambda(em)=443 nm). The mean extraction recovery for PND was 95.2%. The coefficient of variation for intra-assay precision, inter-assay precision and accuracy was less than 10%. The quantification limit with fluorescence detection was 0.010 microg/mL plasma. The method described herein has several advantages over other published methods since it is easy to perform and rapid. It also permits reducing both, solvent use and sample preparation time. The method has been used successfully to assay plasma samples from clinical pharmacokinetic studies. PMID- 16046286 TI - A novel HPLC procedure for detection and quantification of aminoacetone, a precursor of methylglyoxal, in biological samples. AB - Increase in methylglyoxal is thought to be involved in different pathological conditions. Deamination of aminoacetone by semicarbazide-sensitive amine oxidase (SSAO) leads to production of methylglyoxal. We have synthesized aminoacetone and developed a novel HPLC procedure for its quantitative determination. The urinary excretion of aminoacetone is approximately 20-30 microg/mouse/day, and the concentration is about 0.5 microg/g in mouse liver and small intestine. SSAO inhibitor increases aminoacetone levels in both tissues and urines. Results confirm that aminoacetone is an endogenous substrate for SSAO. However, data also indicate that deamination is not the only catabolic pathway for aminoacetone. PMID- 16046287 TI - An improved HPLC method for determination of carotenoids in human serum. AB - An HPLC method was developed to determine the various carotenoids in human serum. A C-30 column and a mobile phase of 100% methanol (A) and 100% methylene chloride (B) with the following gradient elution were used: 90% A and 10% B in the beginning, maintained for 5 min, decreased to 78% A at 15 min, 62% A at 30 min, 52% A at 40 min, 41% A at 50 min, 38% A at 55 min, maintained for 3 min, and returned to 100% A at 65 min. A total of 21 carotenoids, including all-trans forms of lutein, zeaxanthin, alpha-cryptoxanthin, beta-cryptoxanthin, alpha carotene, beta-carotene and lycopene, as well as their 14 cis-isomers were resolved within 51 min at a flow rate of 1.0 mL/min and detection at 476 nm. all trans-beta-Carotene was found to be present in highest amount (256.3-864.2 ng/mL), followed by all-trans-lycopene (64.4-569.2 ng/mL), all-trans-lutein (137.9-450.3 ng/mL), all-trans-alpha-cryptoxanthin (55.7-188.2 ng/mL), all-trans beta-cryptoxanthin (43.1-134.5 ng/mL), all-trans-alpha-carotene (20.0-122.1 ng/mL) and all-trans-zeaxanthin (9.1-21.3 ng/mL). Similar trend was observed for cis-isomers of carotenoids. PMID- 16046288 TI - High-throughput analysis of vitamin C in human plasma with the use of HPLC with monolithic column and UV-detection. AB - Vitamin C plays a central role in the body. One of its important functions is its role as an antioxidant, and accurate measurements are important for interpretations of this role. However, its reactive nature and instability complicates the assessment, especially in biological samples. A high-throughput chromatographic method using monolithic column and UV-detection was developed for the assessment of plasma ascorbic acid and total ascorbic acid. The method showed excellent analytical sensitivity, specificity, precision, recovery and linearity during the validation study. The method was used for the assessment of ascorbic acid and total ascorbic acid during several clinical studies. PMID- 16046289 TI - Identification of human calphoglin-induced phosphoglucomutase phosphorylation in Escherichia coli. AB - Orthologous proteomes, universal protein networks conserved from bacteria to mammals, dictate the core functions of cells. To isolate mammalian protein sequences that interact with bacterial signaling proteins, a BLASTP genome search was performed using catalytic domains of bacterial phosphoryl-transfer enzymes as probes. A [32P]phosphoryl-transfer assay of these mammalian cDNA-expressing Escherichia coli cells was used to screen proteins retrieved from the database. Here we report that the expression of a human protein, named calphoglin, resulted in a significant increase in the phosphorylation of a 55-kDa protein in E. coli. The phosphorylation of the 55-kDa protein was acid-stable and its isoelectric point was determined to be 5.4. The 55-kDa protein was sequentially purified from an E. coli extract using three chromatography and two-dimensional polyacrylamide gel electrophoresis. Finally, the 55-kDa protein was purified 830-fold to homogeneity and the N-terminal amino acid sequence was analyzed. The sequence obtained, AIHNRAGQPAQQ, was identical to the N-terminal amino acids of E. coli phosphoglucomutase (PGM). This method may be applicable to the detection and analysis of other orthologous proteomes. PMID- 16046290 TI - ACCF/AHA clinical competence statement on cardiac imaging with computed tomography and magnetic resonance. PMID- 16046291 TI - Proopiomelanocortin-deficient mice are hypersensitive to the adverse metabolic effects of glucocorticoids. AB - Congenital lack of proopiomelanocortin (POMC) causes obesity and glucocorticoid deficiency. The responses of Pomc-/- and wild-type mice to the administration of corticosterone were compared. In study 1, mice were given corticosterone supplemented water (CORT) for 10 days, resulting in plasma CORT levels within the physiological range, with partial suppression of hypothalamic corticotropin releasing hormone expression to a similar degree between genotypes. Body weight, fat mass, and food intake increased in CORT-treated Pomc-/- but not wild-type mice. CORT increased plasma insulin levels 50-fold in Pomc-/- versus 14-fold in wild-type mice (P < 0.01) and increased hypothalamic agouti-related protein (AgRP) expression by more than 200% in Pomc-/- versus 40% in wild type (P < 0.05). In study 2, mice were given CORT from weaning, and Pomc-/- but not wild type mice developed hyperglycemia, ketonuria, and hepatic steatosis by 8-12 weeks. Thus, Pomc-/- mice are hypersensitive to the adverse metabolic effects of glucocorticoids. Additionally, as the levels of plasma CORT achieved, especially in study 1, were not grossly supraphysiological, we conclude that glucocorticoid deficiency may afford Pomc-/- mice some protection from the full adverse consequences of melanocortin deficiency. This may occur through a mechanism involving the suppression of AgRP by the hypoadrenal state. PMID- 16046292 TI - Reduction of macrophage infiltration and chemoattractant gene expression changes in white adipose tissue of morbidly obese subjects after surgery-induced weight loss. AB - In human obesity, the stroma vascular fraction (SVF) of white adipose tissue (WAT) is enriched in macrophages. These cells may contribute to low-grade inflammation and to its metabolic complications. Little is known about the effect of weight loss on macrophages and genes involved in macrophage attraction. We examined subcutaneous WAT (scWAT) of 7 lean and 17 morbidly obese subjects before and 3 months after bypass surgery. Immunomorphological changes of the number of scWAT-infiltrating macrophages were evaluated, along with concomitant changes in expression of SVF-overexpressed genes. The number of scWAT-infiltrating macrophages before surgery was higher in obese than in lean subjects (HAM56+/CD68+; 22.6 +/- 4.3 vs. 1.4 +/- 0.6%, P < 0.001). Typical "crowns" of macrophages were observed around adipocytes. Drastic weight loss resulted in a significant decrease in macrophage number (-11.63 +/- 2.3%, P < 0.001), and remaining macrophages stained positive for the anti-inflammatory protein interleukin 10. Genes involved in macrophage attraction (monocyte chemotactic protein [MCP]-1, plasminogen activator urokinase receptor [PLAUR], and colony stimulating factor [CSF]-3) and hypoxia (hypoxia-inducible factor-1alpha [HIF 1alpha]), expression of which increases in obesity and decreases after surgery, were predominantly expressed in the SVF. We show that improvement of the inflammatory profile after weight loss is related to a reduced number of macrophages in scWAT. MCP-1, PLAUR, CSF-3, and HIF-1alpha may play roles in the attraction of macrophages in scWAT. PMID- 16046293 TI - Donor islet endothelial cells participate in formation of functional vessels within pancreatic islet grafts. AB - Pancreatic islet transplantation has emerged as a therapy for type 1 diabetes and is today performed using both freshly isolated and cultured islets. Islet blood vessels are disrupted during islet isolation; therefore, proper revascularization of the transplanted islets is of great importance for islet graft function and survival. We have studied intraislet endothelial cells after islet isolation, during islet culture, and following islet transplantation. By isolating islets from the transgenic Tie2-GFP (green fluorescent protein) mouse, characterized by an endothelial cell-specific expression of GFP, living endothelial cells could be studied in intact islets utilizing two-photon laser-scanning microscopy (TPLSM). Intraislet endothelial cells were found to survive islet transplantation but to rapidly disappear during islet culture. By transplanting freshly isolated Tie2 GFP islets and applying a novel ex vivo model for simultaneous perfusion and TPLSM imaging of the graft-bearing kidneys, GFP fluorescent endothelial cells were found to extensively contribute to vessels within the islet graft vasculature. Real-time imaging of the flow through the islet graft vasculature confirmed that the donor-derived vessels were functionally integrated. Hence, intraislet endothelial cells have the capability of participating in revascularization of pancreatic islets subsequent to transplantation. Therefore, preservation of intraislet endothelial cell mass may improve long-term graft function. PMID- 16046294 TI - Mechanisms of compensatory beta-cell growth in insulin-resistant rats: roles of Akt kinase. AB - The physiological mechanisms underlying the compensatory growth of beta-cell mass in insulin-resistant states are poorly understood. Using the insulin-resistant Zucker fatty (fa/fa) (ZF) rat and the corresponding Zucker lean control (ZLC) rat, we investigated the factors contributing to the age-/obesity-related enhancement of beta-cell mass. A 3.8-fold beta-cell mass increase was observed in ZF rats as early as 5 weeks of age, an age that precedes severe insulin resistance by several weeks. Closer investigation showed that ZF rat pups were not born with heightened beta-cell mass but developed a modest increase over ZLC rats by 20 days that preceded weight gain or hyperinsulinemia that first developed at 24 days of age. In these ZF pups, an augmented survival potential of beta-cells of ZF pups was observed by enhanced activated (phospho-) Akt, phospho BAD, and Bcl-2 immunoreactivity in the postweaning period. However, increased beta-cell proliferation in the ZF rats was only detected at 31 days of age, a period preceding massive beta-cell growth. During this phase, we also detected an increase in the numbers of small beta-cell clusters among ducts and acini, increased duct pancreatic/duodenal homeobox-1 (PDX-1) immunoreactivity, and an increase in islet number in the ZF rats suggesting duct- and acini-mediated heightened beta-cell neogenesis. Interestingly, in young ZF rats, specific cells associated with ducts, acini, and islets exhibited an increased frequency of PDX 1+/phospho-Akt+ staining, indicating a potential role for Akt in beta-cell differentiation. Thus, several adaptive mechanisms account for the compensatory growth of beta-cells in ZF rats, a combination of enhanced survival and neogenesis with a transient rise in proliferation before 5 weeks of age, with Akt serving as a potential mediator in these processes. PMID- 16046295 TI - Expression of CD68 and macrophage chemoattractant protein-1 genes in human adipose and muscle tissues: association with cytokine expression, insulin resistance, and reduction by pioglitazone. AB - To examine the role of adipose-resident macrophages in insulin resistance, we examined the gene expression of CD68, a macrophage marker, along with macrophage chemoattractant protein-1 (MCP-1) in human subcutaneous adipose tissue using real time RT-PCR. Both CD68 and MCP-1 mRNAs were expressed in human adipose tissue, primarily in the stromal vascular fraction. When measured in the adipose tissue from subjects with normal glucose tolerance, covering a wide range of BMI (21-51 kg/m2) and insulin sensitivity (S(I)) (0.6-8.0 x 10(-4)min(-1).microU(-1).ml( 1)), CD68 mRNA abundance, which correlated with the number of CD68-positive cells by immunohistochemistry, tended to increase with BMI but was not statistically significant. However, there was a significant inverse relation between CD68 mRNA and S(I) (r=-0.55, P=0.02). In addition, there was a strong positive relationship among adipose tissue CD68 mRNA, tumor necrosis factor-alpha (TNF-alpha) secretion in vitro (r=0.79, P<0.005), and plasma interleukin-6 (r=0.67, P < 0.005). To determine whether improving S(I) in subjects with impaired glucose tolerance (IGT) was associated with decreased CD68 expression, IGT subjects were treated for 10 weeks with pioglitazone or metformin. Pioglitazone increased S(I) by 60% and in the same subjects reduced both CD68 and MCP-1 mRNAs by >50%. Furthermore, pioglitazone resulted in a reduction in the number of CD68-positive cells in adipose tissue and reduced plasma TNF-alpha. Metformin had no effect on any of these measures. Thus, treatment with pioglitazone reduces expression of CD68 and MCP-1 in adipose tissue, apparently by reducing macrophage numbers, resulting in reduced inflammatory cytokine production and improvement in S(I). PMID- 16046296 TI - Insulin resistance and type 2 diabetes in high-fat-fed mice are linked to high glycotoxin intake. AB - Dietary advanced glycosylation end products (AGEs) have been linked to insulin resistance in db/db(++) mice. To test whether dietary AGEs play a role in the progression of insulin resistance in normal mice fed high-fat diets, normal C57/BL6 mice were randomly assigned to high-fat diets (35% g fat), either high (HAGE-HF group; 995.4 units/mg AGE) or low (by 2.4-fold LAGE-HF group; 329.6 units/mg AGE) in AGE content for 6 months. Age-matched C57/BL6 and db/db(++) mice fed regular diet (5% g fat, 117.4 units/mg AGE) served as controls. After 6 months, 75% of HAGE-HF mice were diabetic and exhibited higher body weight (P < 0.001), fasting glucose (P < 0.001), insulin (P < 0.001), and serum AGEs (P < 0.01) than control mice, while none of the LAGE-HF mice were diabetic despite a similar rise in body weight and plasma lipids. The HAGE-HF group displayed markedly impaired glucose and insulin responses during glucose tolerance tests and euglycemic and hyperglycemic clamps and altered pancreatic islet structure and function compared with those of LAGE-HF mice, in which findings resembled those of control mice. The HAGE-HF group had more visceral fat (by two- and fourfold) and more AGE-modified fat (by two- and fivefold) than LAGE-HF and control mice, respectively. In the HAGE-HF group, plasma 8-isoprostane was higher (P < 0.01) and adiponectin lower (P < 0.001) than control mice, while in the LAGE HF group, these were more modestly affected (P < 0.05). These results demonstrate that the development of insulin resistance and type 2 diabetes during prolonged high-fat feeding are linked to the excess AGEs/advanced lipoxidation end products inherent in fatty diets. PMID- 16046297 TI - Carnosine as a protective factor in diabetic nephropathy: association with a leucine repeat of the carnosinase gene CNDP1. AB - The risk of diabetic nephropathy is partially genetically determined. Diabetic nephropathy is linked to a gene locus on chromosome 18q22.3-q23. We aimed to identify the causative gene on chromosome 18 and to study the mechanism by which the product of this gene could be involved in the development of diabetic nephropathy. DNA polymorphisms were determined in 135 case (diabetic nephropathy) and 107 control (diabetes without nephropathy) subjects. The effect of carnosine on the production of extracellular matrix components and transforming growth factor-beta (TGF-beta) after exposure to 5 and 25 mmol/l d-glucose was studied in cultured human podocytes and mesangial cells, respectively. A trinucleotide repeat in exon 2 of the CNDP1 gene, coding for a leucine repeat in the leader peptide of the carnosinase-1 precursor, was associated with nephropathy. The shortest allelic form (CNDP1 Mannheim) was more common in the absence of nephropathy (P = 0.0028, odds ratio 2.56 [95% CI 1.36-4.84]) and was associated with lower serum carnosinase levels. Carnosine inhibited the increased production of fibronectin and collagen type VI in podocytes and the increased production of TGF-beta in mesangial cells induced by 25 mmol/l glucose. Diabetic patients with the CNDP1 Mannheim variant are less susceptible for nephropathy. Carnosine protects against the adverse effects of high glucose levels on renal cells. PMID- 16046298 TI - Regulation of renal lipid metabolism, lipid accumulation, and glomerulosclerosis in FVBdb/db mice with type 2 diabetes. AB - Diabetic kidney disease has been associated with the presence of lipid deposits, but the mechanisms for the lipid accumulation have not been fully determined. In the present study, we found that db/db mice on the FVB genetic background with loss-of-function mutation of the leptin receptor (FVB-Lepr(db) mice or FVBdb/db) develop severe diabetic nephropathy, including glomerulosclerosis, tubulointerstitial fibrosis, increased expression of type IV collagen and fibronectin, and proteinuria, which is associated with increased renal mRNA abundance of transforming growth factor-beta, plasminogen activator inhibitor-1, and vascular endothelial growth factor. Electron microscopy demonstrates increases in glomerular basement membrane thickness and foot process (podocyte) length. We found that there is a marked increase in neutral lipid deposits in glomeruli and tubules by oil red O staining and biochemical analysis for cholesterol and triglycerides. We also detected a significant increase in the renal expression of adipocyte differentiation-related protein (adipophilin), a marker of cytoplasmic lipid droplets. We examined the expression of sterol regulatory element-binding protein (SREBP)-1 and -2, transcriptional factors that play an important role in the regulation of fatty acid, triglyceride, and cholesterol synthesis. We found significant increases in SREBP-1 and -2 protein levels in nuclear extracts from the kidneys of FVBdb/db mice, with increases in the mRNA abundance of acetyl-CoA carboxylase, fatty acid synthase, and 3-hydroxy 3-methylglutaryl-CoA reductase, which mediates the increase in renal triglyceride and cholesterol content. Our results indicate that in FVBdb/db mice, renal triglyceride and cholesterol accumulation is mediated by increased activity of SREBP-1 and -2. Based on our previous results with transgenic mice overexpressing SREBP-1 in the kidney, we propose that increased expression of SREBPs plays an important role in causing renal lipid accumulation, glomerulosclerosis, tubulointerstitial fibrosis, and proteinuria in mice with type 2 diabetes. PMID- 16046299 TI - Association of common variation in the HNF1alpha gene region with risk of type 2 diabetes. AB - It is currently unclear how often genes that are mutated to cause rare, early onset monogenic forms of disease also harbor common variants that contribute to the more typical polygenic form of each disease. The gene for MODY3 diabetes, HNF1alpha, lies in a region that has shown linkage to late-onset type 2 diabetes (12q24, NIDDM2), and previous association studies have suggested a weak trend toward association for common missense variants in HNF1alpha with glucose-related traits. Based on genotyping of 79 common SNPs in the 118 kb spanning HNF1alpha, we selected 21 haplotype tag single nucleotide polymorphisms (SNPs) and genotyped them in >4,000 diabetic patients and control subjects from Sweden, Finland, and Canada. Several SNPs from the coding region and 5' of the gene demonstrated nominal association with type 2 diabetes, with the most significant marker (rs1920792) having an odds ratio of 1.17 and a P value of 0.002. We then genotyped three SNPs with the strongest evidence for association to type 2 diabetes (rs1920792, I27L, and A98V) in an additional 4,400 type 2 diabetic and control subjects from North America and Poland and compared our results with those of the original sample and of Weedon et al. None of the results were consistently observed across all samples, with the possible exception of a modest association of the rare (3-5%) A98V variant. These results indicate that common variants in HNF1alpha either play no role in type 2 diabetes, a very small role, or a role that cannot be consistently observed without consideration of as yet unmeasured genetic or environmental modifiers. PMID- 16046300 TI - Physiological increases in uncoupling protein 3 augment fatty acid oxidation and decrease reactive oxygen species production without uncoupling respiration in muscle cells. AB - Decreased uncoupling protein (UCP)3 is associated with insulin resistance in muscle of pre-diabetic and diabetic individuals, but the function of UCP3 remains unclear. Our goal was to elucidate mechanisms underlying the negative correlation between UCP3 and insulin resistance in muscle. We determined effects of physiologic UCP3 overexpression on glucose and fatty acid oxidation and on mitochondrial uncoupling and reactive oxygen species (ROS) production in L6 muscle cells. An adenoviral construct caused a 2.2- to 2.5-fold increase in UCP3 protein. Palmitate oxidation was increased in muscle cells incubated under normoglycemic or hyperglycemic conditions, whereas adenoviral green fluorescent protein infection or chronic low doses of the uncoupler dinitrophenol had no effect. Increased UCP3 did not affect glucose oxidation, whereas dinitrophenol and insulin treatments caused increases. Basal oxygen consumption, assessed in situ using self-referencing microelectrodes, was not significantly affected, whereas dinitrophenol caused increases. Mitochondrial membrane potential was decreased by dinitrophenol but was not affected by increased UCP3 expression. Finally, mitochondrial ROS production decreased significantly with increased UCP3 expression. Results are consistent with UCP3 functioning to facilitate fatty acid oxidation and minimize ROS production. As impaired fatty acid metabolism and ROS handling are important precursors in muscular insulin resistance, UCP3 is an important therapeutic target in type 2 diabetes. PMID- 16046301 TI - Increased p85/55/50 expression and decreased phosphotidylinositol 3-kinase activity in insulin-resistant human skeletal muscle. AB - Insulin resistance is predominantly characterized by decreased insulin-stimulated glucose uptake into skeletal muscle. In the current study, we have assessed various aspects of the phosphatidylinositol (PI) 3-kinase pathway in skeletal muscle biopsies obtained from normal, obese nondiabetic, and type 2 diabetic subjects, before and after a 5-h insulin infusion. We found a highly significant inverse correlation between in vivo insulin sensitivity (as measured by the glucose infusion rate) and increased protein expression of p85/55/50, protein kinase C (PKC)-theta activity, levels of pSer307 insulin receptor substrate (IRS) 1 and p-Jun NH2-terminal kinase (JNK)-1, and myosin heavy chain IIx fibers. Increased basal phosphorylation of Ser307 IRS-1 in the obese and type 2 diabetic subjects corresponds with decrease in insulin-stimulated IRS-1 tyrosine phosphorylation, PI 3-kinase activity, and insulin-induced activation of Akt and, more prominently, PKC-zeta/lambda. In summary, increased expression of the PI 3 kinase adaptor subunits p85/55/50, as well as increased activity of the proinflammatory kinases JNK-1, PKC-theta, and, to a lesser extent, inhibitor of kappaB kinase-beta, are associated with increased basal Ser307 IRS-1 phosphorylation and decreased PI 3-kinase activity and may follow a common pathway to attenuate in vivo insulin sensitivity in insulin-resistant subjects. These findings demonstrate interacting mechanisms that can lead to impaired insulin-stimulated PI 3-kinase activity in skeletal muscle from obese and type 2 diabetic subjects. PMID- 16046302 TI - Preconditioning the diabetic heart: the importance of Akt phosphorylation. AB - Conflicting evidence exists whether diabetic myocardium can be protected by ischemic preconditioning (IPC). The phosphatidylinositol 3-kinase (PI3K)-Akt pathway is important in IPC. However, components of this cascade have been found to be defective in diabetes. We hypothesize that IPC in diabetic hearts depends on intact signaling through the PI3K-Akt pathway to reduce myocardial injury. Isolated perfused Wistar (normal) and Goto-Kakizaki (diabetic) rat hearts were subjected to 1) 35 min of regional ischemia and 120 min of reperfusion with infarct size determined; 2) preconditioning (IPC) using 5 min of global ischemia followed by 10 min of reperfusion performed one, two, or three times before prolonged ischemia; or 3) determination of Akt phosphorylation after stabilization or after one and three cycles of IPC. In Wistar rats, one, two, and three cycles of IPC reduced infarct size 44.7 +/- 3.8% (P < 0.05), 31.4 +/- 4.9% (P < 0.01), and 34.3 +/- 6.1% (P < 0.01), respectively, compared with controls (60.7 +/- 4.5%). However, in diabetic rats only three cycles of IPC significantly reduced infarction to 20.8 +/- 2.6% from 46.6 +/- 5.2% in controls (P < 0.01), commensurate with significant Akt phosphorylation after three cycles of IPC. To protect the diabetic myocardium, it appears necessary to increase the IPC stimulus to achieve the threshold for cardioprotection and a critical level of Akt phosphorylation to mediate myocardial protection. PMID- 16046303 TI - Skeletal muscle AMP-activated protein kinase phosphorylation parallels metabolic phenotype in leptin transgenic mice under dietary modification. AB - Leptin augments glucose and lipid metabolism independent of its effect on satiety. Administration of leptin in rodents increases skeletal muscle beta oxidation by activating AMP-activated protein kinase (AMPK). We previously reported that, as hyperleptinemic as obese human subjects, transgenic skinny mice overexpressing leptin in liver (LepTg) exhibit enhanced insulin sensitivity and lipid clearance. To assess skeletal muscle AMPK activity in leptin-sensitive and insensitive states, we examined phosphorylation of AMPK and its target, acetyl CoA carboxylase (ACC), in muscles from LepTg under dietary modification. Here we show that phosphorylation of AMPK and ACC are chronically augmented in LepTg soleus muscle, with a concomitant increase in the AMP-to-ATP ratio and a significant decrease in tissue triglyceride content. Despite preexisting hyperleptinemia, high-fat diet (HFD)-fed LepTg develop obesity, insulin resistance, and hyperlipidemia. In parallel, elevated soleus AMPK and ACC phosphorylation in regular diet-fed LepTg is attenuated, and tissue triglyceride content is increased in those given HFD. Of note, substitution of HFD with regular diet causes a robust recovery of soleus AMPK and ACC phosphorylation in LepTg, with a higher rate of body weight reduction and a regain of insulin sensitivity. In conclusion, soleus AMPK and ACC phosphorylation in LepTg changes in parallel with its insulin sensitivity under dietary modification, suggesting a close association between skeletal muscle AMPK activity and sensitivity to leptin. PMID- 16046304 TI - Insulin and inositol 1,4,5-trisphosphate trigger abnormal cytosolic Ca2+ transients and reveal mitochondrial Ca2+ handling defects in cardiomyocytes of ob/ob mice. AB - Obesity, insulin resistance, and type 2 diabetes are leading causes of heart failure, and defective cellular Ca2+ handling seems to be a fundamental problem in diabetes. Therefore, we studied the effect of insulin on Ca2+ homeostasis in normal, freshly isolated mouse ventricular cardiomyocytes and whether Ca2+ handling was changed in an animal model of obesity and type 2 diabetes, ob/ob mice. Electrically evoked Ca2+ transients were smaller and slower in ob/ob compared with wild-type cardiomyocytes. Application of insulin (6 or 60 nmol/l) increased the amplitude of Ca2+ transients in wild-type cells by approximately 30%, whereas it broadened the transients and triggered extra Ca2+ transients in ob/ob cells. The effects of insulin in ob/ob cells could be reproduced by application of a membrane-permeant inositol trisphosphate (IP3) analog and blocked by a frequently used IP3 receptor inhibitor, 2-aminoethoxydiphenyl borate. In ob/ob cardiomyocytes, insulin increased the IP3 concentration and mitochondrial Ca2+ handling was impaired. In conclusion, we propose a model where insulin increases IP3 in ob/ob cardiomyocytes, which prolongs the electrically evoked Ca2+ release. This, together with an impaired mitochondrial Ca2+ handling, results in insulin-mediated extra Ca2+ transients in ob/ob cardiomyocytes that may predispose for arrhythmias in vivo. PMID- 16046305 TI - beta-cell function in morbidly obese subjects during free living: long-term effects of weight loss. AB - Insulin hypersecretion and insulin resistance are physiologically linked features of obesity. We tested whether extreme hypersecretion impairs beta-cell function under free-living conditions and whether major weight loss modifies insulin hypersecretion, insulin sensitivity, and beta-cell function. Plasma glucose, C peptide, and free fatty acid concentrations were measured at hourly intervals during 24 h of normal life (including calorie-standardized meals) in 20 morbidly obese nondiabetic patients (BMI 48.4 +/- 1.7 kg/m2) and 7 nonobese age- and sex matched control subjects; 8 of the obese patients were restudied 6 months and 2 years following biliopancreatic diversion. Insulin secretion was reconstructed from C-peptide levels by deconvolution and related to concurrent glucose levels through a mathematical model incorporating key features of beta-cell function: rate sensitivity, beta-cell glucose sensitivity, and potentiation. Insulin sensitivity (by the euglycemic insulin clamp technique) was reduced by 50% in obese subjects (23.1 +/- 2.5 of obese subjects vs. 52.9 +/- 4.9 micromol.min(-1) . kg(FFM)(-1) of control subjects, means +/- SE, P = 0.0004) as was mean 24-h insulin clearance (median 809 [interquartile range 451] vs. 1,553 [520] ml.min( 1) . m(-2), P < 0.001) due to a 50% reduction in hepatic insulin extraction (P < 0.01). Over 24 h, insulin secretion was doubled in obese subjects (468 nmol [202] in obese subjects vs. 235 [85] of control subjects, P=0.0002). Despite the hypersecretion, beta-cell glucose sensitivity, rate sensitivity, and potentiation were similar in obese and control subjects. Six months postoperatively (weight loss = 33 +/- 3 kg), both insulin hypersecretion (282 nmol [213]) and insulin sensitivity (51.6 +/- 3.7 micromol.min(-1).kg(FFM)(-1)) were normalized. At 2 years (weight loss = 50 +/- 8 kg), insulin sensitivity was supernormal (68.7 +/- 3.3 micromol.min(-1).kg(FFM)(-1)) and insulin secretion was lower than normal (167 nmol [37]) (both P < 0.05 vs. control subjects). In conclusion, severe uncomplicated obesity is characterized by gross insulin hypersecretion and insulin resistance, but the dynamic aspects of beta-cell function are intact. Malabsorptive bariatric surgery corrects both the insulin hypersecretion and the insulin resistance at a time when BMI is still high. With continued weight loss over a 2-year period, moderately obese subjects become supersensitive to insulin and, correspondingly, insulin hyposecretors. PMID- 16046306 TI - Subcutaneous oxyntomodulin reduces body weight in overweight and obese subjects: a double-blind, randomized, controlled trial. AB - This study investigated the effect of subcutaneously administered oxyntomodulin on body weight in healthy overweight and obese volunteers. Participants self administered saline or oxyntomodulin subcutaneously in a randomized, double blind, parallel-group protocol. Injections were self-administered for 4 weeks, three times daily, 30 min before each meal. The volunteers were asked to maintain their regular diet and level of physical exercise during the study period. Subjects' body weight, energy intake, and levels of adipose hormones were assessed at the start and end of the study. Body weight was reduced by 2.3 +/- 0.4 kg in the treatment group over the study period compared with 0.5 +/- 0.5 kg in the control group (P = 0.0106). On average, the treatment group had an additional 0.45-kg weight loss per week. The treatment group demonstrated a reduction in leptin and an increase in adiponectin. Energy intake by the treatment group was significantly reduced by 170 +/- 37 kcal (25 +/- 5%) at the initial study meal (P = 0.0007) and by 250 +/- 63 kcal (35 +/- 9%) at the final study meal (P = 0.0023), with no change in subjective food palatability. Oxyntomodulin treatment resulted in weight loss and a change in the levels of adipose hormones consistent with a loss of adipose tissue. The anorectic effect was maintained over the 4-week period. Oxyntomodulin represents a potential therapy for obesity. PMID- 16046307 TI - Disruption of the gamma-interferon signaling pathway at the level of signal transducer and activator of transcription-1 prevents immune destruction of beta cells. AB - beta-cells under immune attack are destroyed by the aberrant activation of key intracellular signaling cascades. The aim of the present study was to evaluate the contribution of the signal transducer and activator of transcription (STAT)-1 pathway for beta-cell apoptosis by studying the sensitivity of beta-cells from STAT-1 knockout (-/-) mice to immune-mediated cell death in vitro and in vivo. Whole islets from STAT-1-/- mice were completely resistant to interferon (IFN) gamma (studied in combination with interleukin [IL]-1beta)-mediated cell death (92 +/- 4% viable cells in STAT-1-/- mice vs. 56 +/- 3% viable cells in wild-type controls, P < or = 0.001) and had preserved insulin release after exposure to IL 1beta and IFN-gamma. Moreover, analysis of cell death in cytokine-exposed purified beta-cells confirmed that protection was due to absence of STAT-1 in the beta-cells themselves. Deficiency of STAT-1 in islets completely prevented cytokine-induced upregulation of IL-15, interferon inducible protein 10, and inducible nitric oxide synthase transcription but did not interfere with monocyte chemoattractant protein 1 and macrophage inflammatory protein 3alpha expression. In vivo, STAT-1-/- mice were partially resistant to development of diabetes after multiple low-dose streptozotocin injections as reflected by mean blood glucose at 12 days after first injection (159 +/- 28 vs. 283 +/- 81 mg/dl in wild-type controls, P < or = 0.05) and diabetes incidence at the end of the follow-up period (39 vs. 73% in wild-type controls, P < or = 0.05). In conclusion, the present results indicate that STAT-1 is a crucial transcription factor in the process of IFN-gamma-mediated beta-cell death and the subsequent development of immune-mediated diabetes. PMID- 16046308 TI - Role of insulin secretion and sensitivity in the evolution of type 2 diabetes in the diabetes prevention program: effects of lifestyle intervention and metformin. AB - Insulin resistance and beta-cell dysfunction, two factors central to the pathogenesis of type 2 diabetes, were studied in relation to the development of diabetes in a group of participants with impaired glucose tolerance in the Diabetes Prevention Program (DPP) at baseline and after specific interventions designed to prevent diabetes. Participants were randomly assigned to placebo (n = 1,082), metformin (850 mg twice a day) (n = 1,073), or intensive lifestyle intervention (n = 1,079). The diabetes hazard rate was negatively associated with baseline insulin sensitivity (hazard rate ratio = 0.62-0.94 per SD difference, depending on treatment group and measure of sensitivity) and with baseline insulin secretion (hazard rate ratio = 0.57-0.76 per SD). Improvements in insulin secretion and insulin sensitivity were associated with lower hazard rates in all treatment arms (hazard rate ratio = 0.46-0.95 per SD increase and 0.29-0.79 per SD increase, respectively). In multivariate models that included the three metabolic variables (changes in body weight, insulin sensitivity, and insulin secretion) each significantly and independently predicted progression to diabetes when adjusted for the other two variables. The intensive lifestyle intervention, which elicited the greatest reduction in diabetes incidence, produced the greatest improvement in insulin sensitivity and the best preservation of beta cell function after 1 year, whereas the placebo group, which had the highest diabetes incidence, had no significant change in insulin sensitivity and beta cell function after 1 year. In the metformin group, diabetes risk, insulin sensitivity, and beta-cell function at 1 year were intermediate between those in the intensive lifestyle and placebo groups. In conclusion, higher insulin secretion and sensitivity at baseline and improvements in response to treatment were associated with lower diabetes risk in the DPP. The better preventive effectiveness of intensive lifestyle may be due to improved insulin sensitivity concomitant with preservation of beta-cell function. PMID- 16046309 TI - Compromised arterial function in human type 2 diabetic patients. AB - Diabetes is associated with a perturbation of signaling pathways in vascular tissue, which causes vasomotor dysfunction such as hypertension and accelerated atherosclerosis. In the present study, the mechanisms of vasomotor dysfunction, Akt (Thr308 and Ser473) phosphorylation and expression of endothelial NO (nitric oxide) synthase, and inducible NO synthase were investigated in human diabetic internal mammary arteries. The phospho-Akt (Thr308) level in arteries from diabetic patients was reduced to about one-half of the level in nondiabetic patients, suggesting impaired insulin signaling in human diabetic vascular tissue. Augmented vasoconstriction was observed in diabetic arteries, due in part to deficiency of basal and stimulated NO production. This correlated with decreased endothelial NO synthase expression and activity in diabetic vessels. The sensitivity of diabetic vessels to the NO donor, sodium nitroprusside, was reduced as well, suggesting that NO breakdown and/or decreased sensitivity of smooth muscle to NO are also responsible for abnormal vasoconstriction. In addition, the abnormal vasoconstriction in diabetic vessels was not completely abolished in the presence of Nomega-nitro-L-arginine methyl ester, revealing that NO-independent mechanisms also contribute to vasomotor dysfunction in diabetes. In conclusion, diabetes downregulates the Akt-signaling pathway and compromises human arterial function through a decrease in NO availability as well as through NO-independent mechanisms. PMID- 16046310 TI - 3,4-Dideoxyglucosone-3-ene induces apoptosis in renal tubular epithelial cells. AB - Diabetes complications are caused by hyperglycemia. Hyperglycemia results in increased concentrations of glucose degradation products. The study of peritoneal dialysis solution biocompatibility has highlighted the adverse effects of glucose degradation products. Recently, 3,4-dideoxyglucosone-3-ene (3,4-DGE) has been identified as the most toxic glucose degradation product in peritoneal dialysis fluids. Its role in renal pathophysiology has not been addressed. 3,4-DGE induces apoptosis in murine renal tubular epithelial cells in a dose- and time-dependent manner. Peak apoptosis is observed after 72 h of culture. The lethal concentration range is 25-50 micromol/l. 3,4-DGE results in Bax oligomerization, release of cytochrome c from mitochondria, activation of caspases-9 and -3, and Bid proteolysis. Apoptosis induced by 3,4-DGE is caspase dependent and could be prevented by the broad-spectrum caspase inhibitor zVAD-fmk (Z-Val-Ala-DL-Asp fluoromethylketone) and by specific inhibitors of caspases-2, -8, and -9. However, caspase inhibition did not prevent eventual cell death. In contrast, antagonism of Bax by a Ku-70-derived peptide or antisense oligonucleotides prevented both apoptosis and cell death. In conclusion, 3,4-DGE promotes apoptosis of cultured renal parenchymal cells by a Bax- and caspase-dependent mechanism. A role for 3,4-DGE in diabetes complications in the kidney and in the modulation of residual renal function in peritoneal dialysis should be further explored. PMID- 16046311 TI - Platelet function profiles in patients with type 2 diabetes and coronary artery disease on combined aspirin and clopidogrel treatment. AB - To assess platelet function profiles in diabetic and nondiabetic patients on aspirin and clopidogrel therapy, two patient populations were included to investigate the 1) acute effects of a 300-mg clopidogrel loading dose (group 1, n = 52) and 2) long-term effects of clopidogrel (group 2, n = 120) on platelet function in diabetic compared with nondiabetic patients already on aspirin treatment. Patients were stratified according to the presence of type 2 diabetes. Platelet aggregation was assessed using light transmittance aggregometry (groups 1 and 2). Platelet activation (P-selectin expression and PAC-1 binding) was determined using whole-blood flow cytometry (group 2). Clopidogrel response was also assessed. In group 1, platelet aggregation was significantly increased in diabetic (n = 16) compared with nondiabetic (n = 36) patients at baseline and up to 24 h following a 300-mg loading dose (P = 0.005). In group 2, platelet aggregation and activation were increased in diabetic (n = 60) compared with nondiabetic (n = 60) subjects (P < 0.05 for all platelet function assays). Diabetic subjects had a higher number of clopidogrel nonresponders (P = 0.04). Diabetic patients have increased platelet reactivity compared with nondiabetic subjects on combined aspirin and clopidogrel treatment. Reduced sensitivity to antiplatelet drugs may contribute to the increased atherothombotic risk in diabetic patients. PMID- 16046312 TI - Chemical ablation of gastric inhibitory polypeptide receptor action by daily (Pro3)GIP administration improves glucose tolerance and ameliorates insulin resistance and abnormalities of islet structure in obesity-related diabetes. AB - Glucose-dependent insulinotropic polypeptide (gastric inhibitory polypeptide [GIP]) is an important incretin hormone secreted by endocrine K-cells in response to nutrient ingestion. In this study, we investigated the effects of chemical ablation of GIP receptor (GIP-R) action on aspects of obesity-related diabetes using a stable and specific GIP-R antagonist, (Pro3)GIP. Young adult ob/ob mice received once-daily intraperitoneal injections of saline vehicle or (Pro3)GIP over an 11-day period. Nonfasting plasma glucose levels and the overall glycemic excursion (area under the curve) to a glucose load were significantly reduced (1.6-fold; P < 0.05) in (Pro3)GIP-treated mice compared with controls. GIP-R ablation also significantly lowered overall plasma glucose (1.4-fold; P < 0.05) and insulin (1.5-fold; P < 0.05) responses to feeding. These changes were associated with significantly enhanced (1.6-fold; P < 0.05) insulin sensitivity in the (Pro3)GIP-treated group. Daily injection of (Pro3)GIP reduced pancreatic insulin content (1.3-fold; P < 0.05) and partially corrected the obesity-related islet hypertrophy and beta-cell hyperplasia of ob/ob mice. These comprehensive beneficial effects of (Pro3)GIP were reversed 9 days after cessation of treatment and were independent of food intake and body weight, which were unchanged. These studies highlight a role for GIP in obesity-related glucose intolerance and emphasize the potential of specific GIP-R antagonists as a new class of drugs for the alleviation of insulin resistance and treatment of type 2 diabetes. PMID- 16046313 TI - Rosiglitazone reduces glucose-stimulated insulin secretion rate and increases insulin clearance in nondiabetic, insulin-resistant individuals. AB - Compensatory hyperinsulinemia permitting insulin-resistant individuals to maintain normal glucose tolerance is associated with a left shift in the glucose stimulated insulin secretion rate (GS-ISR) dose-response curve and decrease in the insulin metabolic clearance rate (I-MCR). To see whether these changes would reverse with improvement in insulin sensitivity, 14 nondiabetic insulin-resistant subjects received rosiglitazone for 12 weeks (4 mg daily for 4 weeks and then 8 mg daily for 8 weeks). Insulin-mediated glucose uptake was quantified by measuring the steady-state plasma glucose concentration during the insulin suppression test. GS-ISR and I-MCR were determined during a 240-min graded intravenous glucose infusion. I-MCR was also calculated during the insulin suppression test. After rosiglitazone treatment, insulin sensitivity improved with significant fall in steady-state plasma glucose (means +/- SE from 13.5 +/- 0.62 to 9.8 +/- 1.02 mmol/l, P < 0.001). In response, the integrated GS-ISR decreased by 21% (P < 0.001), with a right shift in the dose-response curve. Calculated I-MCR increased by 34% (P = 0.008) during the insulin suppression test and by 21% (P = 0.03) during the graded glucose infusion. In conclusion, enhanced insulin sensitivity in rosiglitazone-treated nondiabetic insulin-resistant individuals was associated with a shift to the right in the GS-ISR dose-response curve and an increase in I-MCR. PMID- 16046314 TI - Glycogen phosphorylase inhibition in type 2 diabetes therapy: a systematic evaluation of metabolic and functional effects in rat skeletal muscle. AB - Inhibition of hepatic glycogen phosphorylase is a promising treatment strategy for attenuating hyperglycemia in type 2 diabetes. Crystallographic studies indicate, however, that selectivity between glycogen phosphorylase in skeletal muscle and liver is unlikely to be achieved. Furthermore, glycogen phosphorylase activity is critical for normal skeletal muscle function, and thus fatigue may represent a major development hurdle for this therapeutic strategy. We have carried out the first systematic evaluation of this important issue. The rat gastrocnemius-plantaris-soleus (GPS) muscle was isolated and perfused with a red cell suspension, containing 3 micromol/l glycogen phosphorylase inhibitor (GPi) or vehicle (control). After 60 min, the GPS muscle was snap-frozen (rest, n = 11 per group) or underwent 20 s of maximal contraction (n = 8, control; n = 9, GPi) or 10 min of submaximal contraction (n = 10 per group). GPi pretreatment reduced the activation of the glycogen phosphorylase a form by 16% at rest, 25% after 20 s, and 44% after 10 min of contraction compared with the corresponding control. AMP-mediated glycogen phosphorylase activation was impaired only at 10 min (by 21%). GPi transiently reduced muscle lactate production during contraction, but other than this, muscle energy metabolism and function remained unaffected at both contraction intensities. These data indicate that glycogen phosphorylase inhibition aimed at attenuating hyperglycaemia is unlikely to negatively impact muscle metabolic and functional capacity. PMID- 16046315 TI - Therapeutic roles of peroxisome proliferator-activated receptor agonists. AB - Peroxisome proliferator-activated receptors (PPARs) play key roles in the regulation of energy homeostasis and inflammation, and agonists of PPARalpha and gamma are currently used therapeutically. Fibrates, first used in the 1970s for their lipid-modifying properties, were later shown to activate PPARalpha. These agents lower plasma triglycerides and VLDL particles and increase HDL cholesterol, effects that are associated with cardiovascular benefit. Thiazolidinediones, acting via PPARgamma, influence free fatty acid flux and thus reduce insulin resistance and blood glucose levels. PPARgamma agonists are therefore used to treat type 2 diabetes. PPARalpha and -gamma agonists also affect inflammation, vascular function, and vascular remodeling. As knowledge of the pleiotropic effects of these agents advances, further potential indications are being revealed, including roles in the management of cardiovascular disease (CVD) and the metabolic syndrome. Dual PPARalpha/gamma agonists (currently in development) look set to combine the properties of thiazolidinediones and fibrates, and they hold considerable promise for improving the management of type 2 diabetes and providing an effective therapeutic option for treating the multifactorial components of CVD and the metabolic syndrome. The functions of a third PPAR isoform, PPARdelta, and its potential as a therapeutic target are currently under investigation. PMID- 16046316 TI - Galanin-like peptide rescues reproductive function in the diabetic rat. AB - Galanin-like peptide (GALP) is expressed in the hypothalamic arcuate nucleus and is regulated by leptin and insulin. Centrally administered GALP stimulates gonadotropin secretion and sexual behavior in the rat. Type 1 diabetes is associated with reduced expression of GALP, as well as an overall decline in reproductive function. We postulated that tonic activity of GALP in the brain is required to sustain normal reproductive activity. To test this hypothesis, we examined whether central (intracerebroventricular) immunoblockade of GALP would reduce sexual behaviors and serum levels of luteinizing hormone (LH) in normal adult male rats. We found that GALP antibody reversibly reduced serum levels of LH and abolished male sexual behaviors (P < 0.05 and 0.001, respectively). Second, we tested whether intracerebroventricular GALP could restore normal plasma LH levels and sexual behavior in diabetic animals. We compared groups of diabetic rats that received intracerebroventricular GALP or vehicle and found that GALP increased serum levels of LH and sexual behavior. Third, we examined whether intracerebroventricular administration of affinity-purified GALP antibody could block the effect of insulin and leptin in reversing the effects of diabetes on LH and sexual behavior. We found that treatment of diabetic animals with insulin and leptin nearly normalized LH levels and sexual behaviors; however, this effect was attenuated by intracerebroventricular administration of GALP antibody (P < 0.05). These observations demonstrate that endogenous GALP provides trophic support to the neuroendocrine reproductive axis, including sexual behavior. PMID- 16046317 TI - A synonymous coding polymorphism in the alpha2-Heremans-schmid glycoprotein gene is associated with type 2 diabetes in French Caucasians. AB - alpha2-Heremans-Schmid glycoprotein (AHSG) is an abundant plasma protein synthesized predominantly in the liver. The AHSG gene, consisting of seven exons and spanning 8.2 kb of genomic DNA, is located at chromosome 3q27, a susceptibility locus for type 2 diabetes and the metabolic syndrome. AHSG is a natural inhibitor of the insulin receptor tyrosine kinase, and AHSG-null mice exhibit significantly enhanced insulin sensitivity. These observations suggested that the AHSG gene is a strong positional and biological candidate for type 2 diabetes susceptibility. Direct sequencing of the AHSG promoter region and exons identified nine common single nucleotide polymorphisms (SNPs) with a minor allele frequency > or =5%. We carried out a detailed genetic association study of the contribution of these common AHSG SNPs to genetic susceptibility of type 2 diabetes in French Caucasians. The major allele of a synonymous coding SNP in exon 7 (rs1071592) presented significant evidence of association with type 2 diabetes (P = 0.008, odds ratio 1.27 [95% CI 1.06-1.52]). Two other SNPs (rs2248690 and rs4918) in strong linkage disequilibrium with rs1071592 showed evidence approaching significance. A haplotype carrying the minor allele of SNP rs1071592 was protective against type 2 diabetes (P = 0.014). However, our analyses indicated that rs1071592 is not associated with the evidence for linkage of type 2 diabetes to 3q27. PMID- 16046318 TI - Association of non-HLA genes with type 1 diabetes autoimmunity. AB - Approximately 50% of the genetic risk for type 1 diabetes is attributable to the HLA region. We evaluated associations between candidate genes outside the HLA region-INS, cytotoxic T-lymphocyte-associated antigen (CTLA)-4, interleukin (IL) 4, IL-4R, and IL-13 and islet autoimmunity among children participating in the Diabetes Autoimmunity Study in the Young (DAISY). Children with persistent islet autoantibody positivity (n = 102, 38 of whom have already developed diabetes) and control subjects (n = 198) were genotyped for single nucleotide polymorphisms (SNPs) in the candidate genes. The INS-23Hph1 polymorphism was significantly associated with both type 1 diabetes (OR = 0.30; 95% CI 0.13-0.69) and persistent islet autoimmunity but in the latter, only in children with the HLA-DR3/4 genotype (0.40; 0.18-0.89). CTLA-4 promoter SNP was significantly associated with type 1 diabetes (3.52; 1.22-10.17) but not with persistent islet autoimmunity. Several SNPs in the IL-4 regulatory pathway appeared to have a predisposing effect for type 1 diabetes. Associations were found between both IL-4R haplotypes and IL-4-IL-13 haplotypes and persistent islet autoimmunity and type 1 diabetes. This study confirms the association between the INS and CTLA-4 loci and type 1 diabetes. Genes involved in the IL-4 regulatory pathway (IL-4, IL-4R, IL-13) may confer susceptibility or protection to type 1 diabetes depending on individual SNPs or specific haplotypes. PMID- 16046319 TI - A large-scale association analysis of common variation of the HNF1alpha gene with type 2 diabetes in the U.K. Caucasian population. AB - HNF1alpha (TCF1) is a key transcription factor that is essential for pancreatic beta-cell development and function. Rare mutations of HNF1alpha cause maturity onset diabetes of the young. A common variant, G319S, private to the Oji-Cree population, predisposes to type 2 diabetes, but the role of common HNF1alpha variation in European populations has not been comprehensively assessed. We determined the linkage disequilibrium and haplotype structure across the HNF1alpha gene region using 29 single nucleotide polymorphisms (SNPs). Eight tagging SNPs (tSNPs) that efficiently capture common haplotypes and the amino acid-changing variant, A98V, were genotyped in 5,307 subjects (2,010 type 2 diabetic case subjects, 1,643 control subjects, and 1,654 members of 521 families). We did not find any evidence of association between the tSNPs or haplotypes and type 2 diabetes. We could exclude odds ratios (ORs) >1.25 for all tSNPs. The rare V98 allele (approximately 3% frequency) showed possible evidence of association with type 2 diabetes (OR 1.23 [95% CI 0.99-1.54], P = 0.07), a result that was supported by meta-analysis of this and published studies (OR 1.31 [1.08-1.59], P = 0.007). Further studies are required to investigate this association, demonstrating the difficulty of defining the role of rare (<5%) alleles in type 2 diabetes risk. PMID- 16046320 TI - Association between common polymorphisms of the proopiomelanocortin gene and body fat distribution: a family study. AB - Rare mutations in the proopiomelanocortin (POMC) gene cause severe early-onset childhood obesity. However, it is unknown whether common variants in POMC are responsible for variation in body weight or fat distribution within the commonly observed range in the population. We tested for association between three polymorphisms spanning the POMC gene and obesity phenotypes in 1,428 members of 248 families. There was significant association between genotypes at the C8246T (P < 0.0001) and C1032G (P = 0.003) polymorphisms and waist-to-hip ratio (WHR) corrected for age, sex, smoking, exercise, and alcohol consumption. Each T allele at C8246T (or G allele at C1032G) was associated with a 0.2-SD-higher WHR in a codominant fashion. When WHR was additionally corrected for BMI, thus providing a measure of body fat distribution throughout the range of BMI, there remained significant evidence for association with both markers that was of similar magnitude and statistical significance. There was no association between genotype at any polymorphism and BMI or plasma leptin level. These data show that genetic variants at the POMC locus influence body fat distribution within the normal range, suggesting a novel role for POMC in metabolic regulation. PMID- 16046321 TI - An uncommonly serious case of an uncommon sport injury. AB - BACKGROUND: A 55 year old man sustained a severe ocular injury when hit by a cricket ball even though he was wearing a helmet. METHODS: A suprachoroidal haemorrhage was drained and dense intravitreal blood was removed. An inferior buckle was applied with the use of intraocular gas. A macular haemorrhage resolved slowly. RESULTS: Despite several surgical procedures over 1.5 years, the final visual acuity of the patient was only 6/60 because of a dense macular scar. CONCLUSIONS: Helmets worn as protection when playing cricket need to be designed better and be of better material. Eye protection should be worn at all levels of play. PMID- 16046322 TI - Isolated oculomotor nerve palsy from minor head trauma. AB - Isolated third cranial nerve palsies in head trauma patients can be the result of direct or indirect damage to the oculomotor nerve. They are usually associated with severe head trauma. We reported a case of isolated oculomotor nerve palsy associated with minor head injury. No initial loss of consciousness was recalled. Computed tomography (CT), magnetic resonance imaging (MRI), and magnetic resonance angiography (MRA) of the brain were normal. Previous reports in the literature were reviewed and the possible mechanism of injury was discussed. Head injuries are commonly seen in sports settings. Our case illustrated that even minor head trauma can cause isolated oculomotor nerve palsy in the absence of abnormal brain imaging findings. PMID- 16046323 TI - Bad hair day... PMID- 16046324 TI - Comment: genetic test available for sports performance. PMID- 16046325 TI - Soccer injury in the lower extremities. AB - Information about soccer injuries is required to develop prevention and rehabilitation programmes. Most soccer injuries occur in the lower extremities. This type of injury is reviewed here. Definitions of injury, injury rate, injury percentage, mechanism of injury, anatomical region of injury, type of injury, and severity of injury are summarised. In each section, a description and summary of the data are provided. Finally, the limitations of the studies and suggestions to improve the investigation of soccer injuries are provided. PMID- 16046326 TI - Reliable change assessment in sport concussion research: a comment on the proposal and reviews of Collie et al. AB - The proposal of Collie et al for determining reliable change in sports medicine and their review of other current procedures did not address mathematical or theoretical aspects. This article attempts to fill this lacuna, in order to examine the validity of their proposal and the suitability of their review. The conclusions drawn are that Collie et al presented their method too carelessly and their review of other methods leaves serious drawbacks and mistakes unnoticed. A scheme is here provided showing which of the current methods can be used in which situation. PMID- 16046327 TI - Body movements on the men's competition mushroom: a three dimensional analysis of circular swings. AB - OBJECTIVES: To develop a method for the three dimensional analysis of body movements and body positions during the performance of circular swings on the competition mushroom, an apparatus used by young gymnasts for pommel horse training. METHODS: Five experienced male gymnasts, all of national level, performed three series of 10 circular swings on the competition mushroom. An optoelectronic instrument was used for the detection of the three dimensional movement of 13 body landmarks. From landmark trajectories, several technical measurements were obtained: diameters of ideal circles of ankles, hips, shoulders; deviation of the ankle diameters from circularity and from the horizontal plane; angle between the shoulder, hip, and ankle. The values were used for a quantitative assessment of performance of the five gymnasts. RESULTS: During the exercise, each ankle should follow a nearly horizontal circular path (deviation from circularity ranged from 3.6% to 6%, deviation from horizontality was 9.4-19.7%), there should be an angle of about 180 degrees at the hips (actual values 146-153 degrees ), and the shoulders should move as little as possible, and only in the lateral plane, without major anteroposterior movements (shoulder movement was 27-31% of ankle movement, hip movement was 16-20%). CONCLUSIONS: The method could help coaches and gymnasts to determine which parts of the body are not repeating a selected movement with sufficient accuracy and to quantify improvements made after a specific training programme. PMID- 16046328 TI - Ocular injury in hurling. AB - OBJECTIVES: To describe the clinical characteristics of ocular injuries sustained in hurling in the south of Ireland and to investigate reasons for non-use of protective headgear and eye wear. METHODS: Retrospective review of the case notes of 310 patients who attended Cork University Hospital or Waterford Regional Hospital between 1 January 1994 and 31 December 2002 with ocular injuries sustained during a hurling match. A confidential questionnaire on reasons for non use of protective headgear and eye wear was completed by 130 players. RESULTS: Hurling related eye injuries occurred most commonly in young men. Fifty two patients (17%) required hospital admission, with hyphaema accounting for 71% of admissions. Ten injuries required intraocular surgical INTERVENTION: retinal detachment repair (5); macular hole surgery (1); repair of partial thickness corneal laceration (1); repair of globe perforation (1); enucleation (1); trabeculectomy for post-traumatic glaucoma (1). Fourteen eyes (4.5%) had a final best corrected visual acuity (BCVA) of <6/12 and six (2%) had BCVA <3/60. In the survey, 63 players (48.5%) reported wearing no protective facemask while playing hurling. Impairment of vision was the most common reason cited for non-use. CONCLUSIONS: Hurling related injury is a significant, and preventable, cause of ocular morbidity in young men in Ireland. The routine use of appropriate protective headgear and faceguards would result in a dramatic reduction in the incidence and severity of these injuries, and should be mandatory. PMID- 16046330 TI - Response of bone metabolism related hormones to a single session of strenuous exercise in active elderly subjects. AB - OBJECTIVE: To evaluate the effect of strenuous exercise on bone metabolism and related hormones in elderly subjects. METHODS: Twenty one active elderly subjects (11 men and 10 women; mean age 73.3 years) showing a mean theoretical Vo2max of 151.4% participated. Concentrations of plasma ionised calcium (iCa), serum intact parathyroid hormone (iPTH), 25-hydroxyvitamin D (25(OH)D), and 1.25-dihydroxy vitamin D3 (1.25(OH)2D3), as well as the bone biochemical markers type I collagen C-telopeptide for bone resorption and osteocalcin and bone alkaline phosphatase for bone formation, were analysed before and after a maximal incremental exercise test. RESULTS: At basal level, iPTH was positively correlated with age (r = 0.56, p < 0.01) and negatively correlated with 25(OH)D (r = -0.50; p < 0.01) and 1.25(OH)2D3 (r = -0.47; p < 0.05). Moreover, 25(OH)D and 1.25(OH)2D3 levels were negatively correlated with age (r = -0.50, p < 0.01 and r = -0.53, p < 0.01, respectively). After exercise, iCa and 25(OH)D decreased (p < 0.001 and p = 0.01, respectively) while iPTH increased (p < 0.001). The levels of 1.25(OH)2D3, bone biochemical markers, haematocrit, and haemoglobin were unchanged. The variations in iCa and 25(OH)D were not related to age and/or sex. The iPTH variation was directly related to basal iPTH levels (p < 0.01) and indirectly related to age. CONCLUSIONS: In active elderly subjects, strenuous exercise disturbed calcium homeostasis and bone related hormones without immediate measurable effect on bone turnover. Although an increase in iPTH could have an anabolic action on bone tissue, our findings from our short term study did not allow us to conclude that such action occurred. PMID- 16046331 TI - Cooling methods used in the treatment of exertional heat illness. AB - OBJECTIVE: To review the different methods of reducing body core temperature in patients with exertional heatstroke. METHODS: The search strategy included articles from 1966 to July 2003 using the databases Medline and Premedline, Embase, Evidence Based Medicine (EBM) reviews, SPORTDiscus, and cross referencing the bibliographies of relevant papers. Studies were included if they contained original data on cooling times or cooling rates in patients with heat illness or normal subjects who were subjected to heat stress. RESULTS: In total, 17 papers were included in the analysis. From the evidence currently available, the most effective method of reducing body core temperature appears to be immersion in iced water, although the practicalities of this treatment may limit its use. Other methods include both evaporative and invasive techniques, and the use of chemical agents such as dantrolene. CONCLUSIONS: The main predictor of outcome in exertional heatstroke is the duration and degree of hyperthermia. Where possible, patients should be cooled using iced water immersion, but, if this is not possible, a combination of other techniques may be used to facilitate rapid cooling. There is no evidence to support the use of dantrolene in these patients. Further work should include a randomised trial comparing immersion and evaporative therapy in heatstroke patients. PMID- 16046332 TI - Injuries in elite motorcycle racing in Japan. AB - OBJECTIVES: To investigate the incidence and pattern of injuries, relative risks, and factors affecting incidence among elite motorcycle competitors in Japan. METHODS: A total of 117 elite motorcycle competitors including 36 road racers, 60 motocross racers, and 21 trial bike riders completed a questionnaire about injuries. RESULTS: Sixty major injuries (25 in road racing, 32 in motocross, and three in trial bike riding) were reported. The most common injuries were fractures (45), followed by ligament injuries (8), dislocations (5), and soft tissue injuries (2). The overall injury rate was 22.4 per 1000 hours, and the death rate was zero. There was no significant correlation between risk of injury and age, experience, or accumulated competition points. CONCLUSIONS: Injury rates in competitions such as road racing and motocross are high, and therefore additional safety measures are needed to protect competitors from injury. PMID- 16046333 TI - Accidental breaches of the doping regulations in sport: is there a need to improve the education of sportspeople? AB - OBJECTIVES: To identify educational needs of elite sportspeople with respect to the doping laws. METHODS: A questionnaire survey of 196 Olympic level sportspeople from the fields of athletics, cycling, rowing, and sailing. The questionnaire addressed the date and source of the last doping educational update, the usefulness of current resources, sources of help, and possible ways of improving the system. The questionnaire also sought to estimate the use of nutritional supplements in these sports. RESULTS: Seventy four (38%) athletes responded to the questionnaire. Over 90% of responders had received a doping educational update in the last six months, and most agreed with the statement "I have received the information I need to avoid getting into trouble with the doping laws". Despite this, more than half of responders agreed with the statements "I should receive reminders more often" and "The authorities should do more to educate sportspeople". In addition, there were four people who admitted taking a banned substance by accident. Forty one (55%) reported taking supplements. The team doctor was the most popular source if information on a substance or product was required, with 62% and 66% of subjects stating that they would contact their team doctor when based in the United Kingdom and abroad respectively. The UK Sports website was often suggested in relation to ways of improving knowledge. CONCLUSIONS: There is a need to alter the educational process, particularly with respect to contingency planning for minor illness. The use of internet based resources for up to date information about banned substances needs to be promoted, and access to the internet improved. The educational needs of team doctors with respect to the doping laws need to be assessed. PMID- 16046334 TI - Effect of aerobic exercise training on mtDNA deletion in soleus muscle of trained and untrained Wistar rats. AB - BACKGROUND: According to the theory of mitochondrial aging, oxidative stress plays a major role in aging and age related degenerative diseases. Since oxygen consumption and reactive oxygen species rate increase during aerobic exercise, we hypothesised that heavy aerobic training could lead to enhanced mitochondrial DNA (mtDNA) deletion in postmitotic tissues, leading in turn to premature aging and degenerative diseases. METHODS: Sixty adult male 2 month old Wistar14848 rats were divided into six equal groups. Two groups were trained for 3 months by running on a treadmill (5 days/week, incline 6 degrees; group 1: 40 m/min, 20 min/day; group 2: 20 m/min, 40 min/day), while two sedentary groups participated in aerobic exercise only at the end of the study (incline 6 degrees; group 3: 40 m/min; group 4: 20 m/min). To control for physical and physiological parameters, two groups of untrained animals were killed at the beginning (group 6) and end (group 5) of the study. Expand long PCR was used to investigate mtDNA deletion in soleus muscle and a sequencing method was used to confirm the mtDNA deletion break point. RESULTS: Our results did not show any mtDNA deletion in untrained rats or in those that underwent moderate training (group 2) We only found mtDNA deletion (4.6 kb) in the soleus muscle of heavily trained rats (group 1). CONCLUSIONS: These results demonstrate that one session of aerobic exercise does not cause mtDNA deletion in skeletal muscle. The difference in results between heavy and moderate aerobic training may be due to low work rate or up-regulation of inducible antioxidant systems in moderate training. PMID- 16046335 TI - "Bounce at the Bell": a novel program of short bouts of exercise improves proximal femur bone mass in early pubertal children. AB - OBJECTIVES: To examine the effects of a simple and inexpensive physical activity intervention on change in bone mass and structure in school aged children. METHODS: Fifty one children (n = 23 boys and 28 girls; mean age 10.1 years) participated in "Bounce at the Bell" which consisted of 10 counter-movement jumps 3x per day (total approximately 3 min/day). Controls were 71 matched children who followed usual school practice. We assessed dietary calcium, physical activity, physical performance, and anthropometry in September and after 8 months of intervention (June). We measured bone mineral content (BMC) and bone area at the lumbar spine, total body, and proximal femur. Proximal femur scans were also analysed for bone geometry and structural strength using the hip structural analysis program. Lean and fat mass (g) were also calculated. RESULTS: Groups were similar at baseline and did not differ in weight, height, total body, lumbar spine, proximal femur, or femoral neck BMC. Control children had a greater increase in adjusted total body BMC (1.4%). Intervention children gained significantly more BMC at the total proximal femur (2%) and the intertrochanteric region (27%). Change in bone structural parameters did not differ between groups. CONCLUSIONS: This novel, easily implemented exercise program, took only a few minutes each day and enhanced bone mass at the weight bearing proximal femur in early pubertal children. A large, randomised study of boys and girls should be undertaken powered to test the effectiveness of Bounce at the Bell in children at different stages of maturity, and in boys and girls independently. PMID- 16046336 TI - Role of exercise stress test in master athletes. AB - BACKGROUND: The effectiveness of cardiovascular screening in minimising the risk of athletic field deaths in master athletes is not known. OBJECTIVE: To evaluate the prevalence and clinical significance of ST segment depression during a stress test in asymptomatic apparently healthy elderly athletes. METHODS: A total of 113 male subjects aged over 60 were studied (79 trained and 34 sedentary); 88 of them (62 trained and 26 sedentary) were followed up for four years (mean 2.16 years for athletes, 1.26 years for sedentary subjects), with a resting 12 lead electrocardiogram (ECG), symptom limited exercise ECG on a cycle ergometer, echocardiography, and 24 hour ECG Holter monitoring. RESULTS: A significant ST segment depression at peak exercise was detected in one athlete at the first evaluation. A further case was seen during the follow up period in a previously "negative" athlete. Both were asymptomatic, and single photon emission tomography and/or stress echocardiography were negative for myocardial ischaemia. The athletes remained symptom-free during the period of the study. One athlete died during the follow up for coronary artery disease: he showed polymorphous ventricular tachycardia during both the exercise test and Holter monitoring, but no significant ST segment depression. CONCLUSIONS: The finding of false positive ST segment depression in elderly athletes, although still not fully understood, may be related to the physiological cardiac remodelling induced by regular training. Thus athletes with exercise induced ST segment depression, with no associated symptoms and/or complex ventricular arrhythmias, and no adverse findings at second level cardiological testing, should be considered free from coronary disease and safe to continue athletic training. PMID- 16046338 TI - Medical survey of female boxing in Italy in 2002-2003. AB - BACKGROUND: Female boxing has been permitted in Italy since 2001. According to the latest Italian laws, athletes applying to become boxers have to pass a pre participation medical examination. OBJECTIVE: To collect novel medical information from the pre-participation visits and mandatory pre-competition and post-competition examinations for all fights involving Italian female boxers in 2002-2003. METHODS: A retrospective study on all official female boxing competitions in Italy from January 2002 to October 2003 was conducted. A prospective study on 28 amateur female boxers was also carried out. RESULTS: Retrospective study: data from 664 examinations were collected. Pre-match examinations were negative. After competitions, 19/645 visits showed some injuries, with mild, soft tissue facial lesions, epistaxis, and hand-wrist problems being the most common. Prospective study: no major lesions were found during the study. One fibroadenoma, one ovarian cyst, and one intramural uterine myoma were found. One boxer was referred to a neurologist because of non-specific electroencephalographic (EEG) abnormalities, which persisted six months later. On a re-admission examination, which was needed because of a contest that was stopped because the referee judged that she was receiving blows to the head that were dangerous, one boxer showed non-specific EEG alterations and nystagmus. A cerebral magnetic resonance imaging scan was normal. She was allowed to participate in competitions again when her EEG returned to normal and clinical signs disappeared. Deviation of the nasal septum was quite common (68%). No major eye injuries were reported. CONCLUSION: Probably because of the correct preventive medical approach, female boxing is much safer than expected, and no major lesions (requiring hospital admission) were reported. Any lesions to the breast and reproductive system could not be considered to be boxing related. PMID- 16046339 TI - Effect of incremental exercise on initiation and movement times in a choice response, whole body psychomotor task. AB - OBJECTIVES: To examine how exercise at moderate and maximal intensities affects performance on a choice response time, whole body psychomotor task. METHODS: Subjects (n = 12) were tested on a three-choice response time task, after rest and after exercise at 70% and 100% maximum power output (W*max). The dependent variables were time to begin forward momentum (initiation time) and time to complete the movement (movement time). Stride time for the first and second strides and number of strides to cover first 1.1 m were also measured. Blood lactate concentrations and heart rate were recorded before and after completion of each psychomotor test. The subjects subjectively assessed the amount of effort that they used to complete the task. RESULTS: Repeated measures analysis of variance showed a significant effect for initiation (F2,22 = 11.47, p < 0.001) and movement times (F2,22 = 14.61, p < 0.001). Post hoc least significant difference (LSD) tests showed that initiation time after exercise at 70% W*max was significantly faster than that in the other two conditions. Speed of initiation after rest was significantly quicker than that after exercise at W*max. For movement time, LSD tests showed that time after maximal exercise was significantly slower than that in the other two conditions. Stride time for the second stride showed a significant effect (F2,22 = 6.20, p < 0.01). LSD tests found that time after exercise at W*max was significantly slower in the other two conditions. Stepwise multiple regression analyses found that the increment of change, from rest, of lactate concentrations could significantly predict the increment of change in initiation (R2 = 0.40) and movement (R2 = 0.50) times. CONCLUSIONS: Exercise affects a whole body task differently from purely cognitive tasks. Central factors are probably more important than peripheral factors. PMID- 16046340 TI - UEFA Champions League study: a prospective study of injuries in professional football during the 2001-2002 season. AB - BACKGROUND: No previous study on adult football involving several different countries has investigated the incidence and pattern of injuries at the highest club competitive level. OBJECTIVE: To investigate the risk exposure, risk of injury, and injury pattern of footballers involved in UEFA Champions League and international matches during a full football season. METHOD: Eleven top clubs (266 players) in five European countries were followed prospectively throughout the season of 2001-2002. Time-lost injuries and individual exposure times were recorded during all club and national team training sessions and matches. RESULTS: A total of 658 injuries were recorded. The mean (SD) injury incidence was 9.4 (3.2) injuries per 1000 hours (30.5 (11.0) injuries per 1000 match hours and 5.8 (2.1) injuries per 1000 training hours). The risk of match injury was significantly higher in the English and Dutch teams than in the teams from France, Italy, and Spain (41.8 (3.3) v 24.0 (7.9) injuries per 1000 hours; p = 0.008). Major injuries (absence > 4 weeks) constituted 15% of all injuries, and the risk of major injury was also significantly higher among the English and Dutch teams (p = 0.04). National team players had a higher match exposure, with a tendency towards a lower training injury incidence than the rest of the players (p = 0.051). Thigh strain was the most common injury (16%), with posterior strains being significantly more common than anterior ones (67 v 36; p < 0.0001). CONCLUSIONS: The risk of injury in European professional football is high. The most common injury is the thigh strain typically involving the hamstrings. The results suggest that regional differences may influence injury epidemiology and traumatology, but the factors involved are unclear. National team players have a higher match exposure, but no higher risk of injury than other top level players. PMID- 16046341 TI - Effects of weight bearing and non-weight bearing exercises on bone properties using calcaneal quantitative ultrasound. AB - OBJECTIVE: This study was designed to investigate bone properties using heel quantitative ultrasound (QUS) in young adults participating in various sports. METHODS: A cross sectional study was performed on Chinese male students (n = 55), aged 18-22 years. Subjects with previous fractures or suffering from any diseases known to affect bone metabolism or taking any medication with such an effect, were not included. The subjects were categorised according to their main sporting activities, including soccer (n = 15) (a high impact, weight bearing exercise), dancing (n = 10) (a low impact, weight bearing exercise), and swimming (n = 15) (non-weight bearing exercise). A sedentary group acted as controls (n = 15). A reproducibility study of the velocity of sound (VOS) and the broadband ultrasound attenuation (BUA) measurement was performed and analysed using the intraclass correlation coefficient (ICC). RESULTS: There was good intra-investigator and inter-investigator agreement (ICC > or = 0.8; p < 0.05) in the measurement of BUA and VOS. No significant differences in BUA and VOS (p > 0.05) were found between the dominant and non-dominant heel. Soccer players (137 +/- 4.3 dB/MHz; 1575 +/- 56 m/s; 544.1 +/- 48.4) and dancers (134.6 +/- 3.7 dB/MHz; 1538 +/- 46 m/s; 503.0 +/- 37.0) had significantly higher BUA, VOS, and stiffness index (SI) scores (p < 0.05), respectively, than swimmers (124.1 +/- 5.1 dB/MHz; 1495 +/- 42 m/s; 423.3 +/- 46.9) and the sedentary control group (119.9 +/- 6.1 dB/MHz; 1452 +/- 41 m/s; 369.9 +/- 46.4). A trend of a significant linear increase with the weight bearing and high impact exercise was revealed in all QUS parameters (p < 0.05). CONCLUSION: This cross sectional study indicated that regular participation in weight bearing exercise in young people might be beneficial for accruing peak bone mass and optimising bone structure. PMID- 16046342 TI - Open label study of intranasal sumatriptan (Imigran) for footballer's headache. AB - OBJECTIVE: To study the efficacy and practicality of treating headache in professional footballers with intranasal sumatriptan. METHODS: An open label drug trial was performed in elite Australian footballers using intranasal sumatriptan (20 mg) treatment for acute headache. The main outcome measures were treatment response at 30 minutes, two hours, and 24 hours using two criteria: (a) initial severity moderate or severe to nil or mild; (b) stricter criteria of initial severity moderate to severe to subsequent nil headache. RESULTS: Thirty eight attacks were analysed. The two hour response showed that 86% of attacks of migraine with aura and all of the attacks of migraine without aura responded to treatment with sumatriptan nasal spray. Complete relief of headache at two hours was reported by 71% of players with migraine with aura and 90% of those without aura. Recurrence rates were generally low, with 0% of migraine headaches and 25% non-migraine attacks recurring at 24 hours. Minor side effects were reported in 28 attacks. CONCLUSIONS: This pilot open label trial suggests that sumatriptan nasal spray may be a valuable, effective, and convenient treatment of headache in professional sport. There are potential risks of this drug that need to be considered. PMID- 16046343 TI - Effects of intra-session concurrent endurance and strength training sequence on aerobic performance and capacity. AB - AIM: To examine the effects of the sequencing order of individualised intermittent endurance training combined with muscular strengthening on aerobic performance and capacity. METHODS: Forty eight male sport students (mean (SD) age 21.4 (1.3) years) were divided into five homogeneous groups according to their maximal aerobic speeds (vV*o2max). Four groups participated in various training programmes for 12 weeks (two sessions a week) as follows: E (n = 10), running endurance training; S (n = 9), strength circuit training; E+S (n = 10) and S+E (n = 10) combined the two programmes in a different order during the same training session. Group C (n = 9) served as a control. All the subjects were evaluated before (T0) and after (T1) the training period using four tests: (1) a 4 km time trial running test; (2) an incremental track test to estimate vV*o2max; (3) a time to exhaustion test (t(lim)) at 100% vV*o2max; (4) a maximal cycling laboratory test to assess V*o2max. RESULTS: Training produced significant improvements in performance and aerobic capacity in the 4 km time trial with interaction effect (p < 0.001). The improvements were significantly higher for the E+S group than for the E, S+E, and S groups: 8.6%, 5.7%, 4.7%, and 2.5% for the 4 km test (p < 0.05); 10.4%, 8.3%, 8.2%, and 1.6% for vV*o2max (p < 0.01); 13.7%, 10.1%, 11.0%, and 6.4% for V*o2max (ml/kg(0.75)/min) (p < 0.05) respectively. Similar significant results were observed for t(lim) and the second ventilatory threshold (%V*o2max). CONCLUSIONS: Circuit training immediately after individualised endurance training in the same session (E+S) produced greater improvement in the 4 km time trial and aerobic capacity than the opposite order or each of the training programmes performed separately. PMID- 16046344 TI - Short term and long term detraining: is there any difference between young-old and old people? AB - OBJECTIVES: To assess the effects of short (six weeks) and long (52 weeks) term detraining on functional fitness in elderly people, and to determine whether these effects differ according to age in elderly people. METHODS: Elderly subjects, aged 60-86 years, completed a nine week multicomponent exercise training programme. They performed the senior fitness test after six and 52 weeks, and the responses of 12 young-old subjects (YO, aged 60-73 years) and nine older subjects (O, aged 74-86 years) were compared. RESULTS: Functional fitness improved during the exercise training period. Short term detraining caused a loss of this improvement in functional performance. Performance on the chair stand test for both YO and O groups and on the up and go and six minute walk tests for the YO group remained significantly higher than before training after six weeks of detraining (p<0.013). Performance in all tests reverted to the pre-training values or lower after 52 weeks of detraining in both groups. In the O group, performances in the six minute walk test and arm curl test were lower than before training (p<0.013). The components of functional fitness most affected by detraining were agility with short term detraining, and aerobic endurance and upper extremity strength with long term detraining. CONCLUSION: Changes in functional capacity after short and long term detraining are affected by age in elderly adults. PMID- 16046345 TI - Effects of six weeks of detraining on retention of functional fitness of old people after nine weeks of multicomponent training. AB - OBJECTIVES: To examine the effects of age on functional fitness after six weeks of detraining. METHODS: Elderly subjects, aged 60-86 years, completed a nine week multicomponent exercise training programme. They performed the senior fitness test every two weeks during the six week detraining period, and the responses of 12 young-old subjects (YO, aged 60-73 years) and nine older subjects (O, aged 74 86 years) were compared. RESULTS: Functional fitness improved during the exercise training period. Performances in the chair stand and six minute walk for the O group had significantly declined compared with post-training values after two weeks of detraining (p<0.01), whereas there were no significant changes in the YO group. Scores on the functional fitness tests declined further between two and four weeks of detraining in both of the groups (p<0.01). In the YO group, there were significant losses in performance on the chair stand, chair sit and reach, and six minute walk tests, and in the O group on the chair stand and up and go tests after six weeks of detraining compared with after four weeks of detraining (p<0.01). The components of functional fitness most affected by detraining were lower extremity flexibility after two and four weeks of detraining, and agility/dynamic balance after six weeks of detraining. CONCLUSION: Changes in lower extremity flexibility, up and go, and six minute walk performances in response to six weeks of detraining are affected by age in elderly adults. PMID- 16046346 TI - Surgical repair of complete proximal hamstring tendon ruptures in water skiers and bull riders: a report of four cases and review of the literature. AB - OBJECTIVE: Hamstring strains are one of the most common muscle strains in athletes; however, complete rupture of the proximal hamstring origin is rare and results from significant trauma. The objective of this paper is to present our experience of management of complete ruptures where surgical repair resulted in good results in both acute and delayed cases. METHODS: Two water skiers and two bull riders sustained complete rupture of the proximal origin of the hamstring muscles. All underwent repair of the hamstring origin and sciatic nerve neurolysis. A post operative hamstring rehabilitation programme was instituted. Regular follow up was performed at 2, 3, 6, 9, and 12 months. RESULTS: At a minimum final follow up of 12 months all patients had regained functional knee flexion strength with no pain and a near normal range of knee flexion. All four individuals were able to return to their previous line of work and three were able to return to their pre-injury level of sport. CONCLUSION: Complete rupture of the hamstring origin is a potentially devastating sports injury that has implications affecting the individual's activities of daily living as well as potential as a sportsperson. Surgical repair restores the distorted anatomy, allows early functional rehabilitation, and avoids the potential debilitating neurological problem of gluteal sciatica. PMID- 16046347 TI - Incidence of serious injury and death during sport and recreation activities in Victoria, Australia. AB - BACKGROUND: Participation in sport and recreation is widely encouraged for general good health and the prevention of some non-communicable diseases. However, injury is a significant barrier to participation, and safety concerns are a factor in the decision to participate. An understanding of the sport/recreation activities associated with serious injury is useful for informing physical activity choices and for setting priorities for the targeting of injury prevention efforts. OBJECTIVES: To describe the epidemiology of serious injuries sustained in sport/recreation activities by adults in Victoria, Australia. METHODS: The Victorian State Trauma Registry and the National Coroner's Information Service were used to identify and describe sport/recreation related serious injuries, including deaths, occurring during the period July 2001 to June 2003. Age adjusted rates of serious injury and death were calculated using participation figures for each sport and general population data. RESULTS: There were 150 cases of serious injury and 48 deaths. The rates of serious injury and death were 1.8 and 0.6 per 100,000 participants per year respectively. Motor, power boat, and equestrian sports had the highest rates of serious injury. Most deaths were due to drowning. CONCLUSION: Although the risk of serious injury through sport/recreation participation is low, motor, power boat, and equestrian sports should be priorities for further research into injury prevention. Most sport/recreation related deaths are due to drowning, highlighting this area for prevention efforts. PMID- 16046349 TI - High VO2MAX with no history of training is due to high blood volume: an alternative explanation. PMID- 16046350 TI - Head injuries. PMID- 16046351 TI - Heading in football. Part 1: development of biomechanical methods to investigate head response. AB - OBJECTIVES: There has been growing controversy regarding long term effects of repeated low severity head impacts such as when heading a football. However, there are few scientific data substantiating these concerns in terms of the biomechanical head response to impact. The present study aimed to develop a research methodology to investigate the biomechanical response of human subjects during intentional heading and identify strategies for reducing head impact severity. METHODS: A controlled laboratory study was carried out with seven active football players, aged 20-23 and of average stature and weight. The subjects were fitted with photographic targets for kinematic analysis and instrumented to measure head linear/angular accelerations and neck muscle activity. Balls were delivered at two speeds (6 m/s and 8 m/s) as the subjects executed several specific forward heading manoeuvres in the standing position. Heading speeds up to 11 m/s were seen when the head closing speed was considered. One subject demonstrating averaged flexion-extension muscle activity phased with head acceleration data and upper torso kinematics was used to validate a biofidelic 50th percentile human model with a detailed head and neck. The model was exercised under ball incoming speeds of 6-7 m/s with parameter variations including torso/head alignment, neck muscle tensing, and follow through. The model output was subsequently compared with additional laboratory tests with football players (n = 3). Additional heading scenarios were investigated including follow through, non-active ball impact, and non-contact events. Subject and model head responses were evaluated with peak linear and rotational accelerations and maximum incremental head impact power. RESULTS: Modelling of neck muscle tensing predicted lower head accelerations and higher neck loads whereas volunteer head acceleration reductions were not consistent. Modelling of head-torso alignment predicted a modest reduction in volunteer head accelerations. Exaggerated follow through while heading reduced volunteer head accelerations modestly. CONCLUSION: Biomechanical methods were developed to measure head impact response. Changing the biomechanics of currently accepted heading techniques will have inconsistent benefits towards the reduction of head loading. Furthermore, mathematical modelling suggested an increased risk of neck loads with one alternative technique. No consistent recommendations can be made on the basis of the current study for altering heading techniques to reduce impact severity. PMID- 16046352 TI - Heading in football. Part 2: biomechanics of ball heading and head response. AB - OBJECTIVES: Controversy surrounding the long term effects of repeated impacts from heading has raised awareness among the public and the medical community. However, there is little information about the human response to the impacts and what measures can be taken to alter their effect. The objective of the current study was to gain a better understanding of heading biomechanics through the implementation of a numerical model and subsequent investigation of parameters related to heading technique and ball characteristics. METHODS: A controlled laboratory study was carried out with seven active football players, aged 20-23 years who underwent medical screening and were instrumented with accelerometers mounted in bite plates and electromyographic electrodes on the major neck muscle groups. Balls were delivered at two speeds (6 m/s and 8 m/s) as the subjects demonstrated several specific heading manoeuvres. Photographic targets were tracked via high speed video to measure heading kinematics. One subject demonstrating reasonably averaged flexion-extension muscle activity phased with head acceleration data and upper torso kinematics was used to validate a biofidelic 50th percentile human numerical model with detailed representation of the head and neck. RESULTS: Heading kinematics and subject responses were used with a detailed numerical model to simulate impact biomechanics for a baseline heading scenario. Changes to heading techniques and ball characteristics which mitigated head impact response were identified. CONCLUSION: A numerical model combined with biomechanical measurement techniques is an important tool for parametric investigation of strategies to reduce head impact severity via changes in heading technique or the physical properties of the ball. PMID- 16046353 TI - A six year prospective study of the incidence and causes of head and neck injuries in international football. AB - OBJECTIVE: To identify those risk factors that have the greatest impact on the incidence of head and neck injuries in international football. METHOD: A case control study of players sustaining head and neck injuries during 20 FIFA tournaments (men and women) from 1998 to 2004. Video recordings of incidents were used to identify a range of parameters associated with the incidents. Team physicians provided medical reports describing the nature of each injury. chi2 tests (p< or =0.01) and 95% confidence intervals were used to assess differences in distribution and incidence of injury, respectively. RESULTS: In total, 248 head and neck injuries were recorded of which 163 were identified and analysed on video sequences. The commonest injuries were contusions (53%), lacerations (20%), and concussions (11%). The incidence of all head and neck injuries was 12.5/1000 player hours (men 12.8, women 11.5) and 3.7 for lost-time injuries (men 3.5, women 4.1). The commonest causes of injury involved aerial challenges (55%) and the use of the upper extremity (33%) or head (30%). The unfair use of the upper extremity was significantly more likely to cause an injury than any other player action. Only one injury (a neck muscle strain) occurred as a result of heading the ball throughout the 20 tournaments equivalent to 0.05 injuries/1000 player hours. CONCLUSIONS: Players' actions most likely to cause a head or neck injury were the use of the upper extremity or the head but in the majority of cases these challenges were deemed to be fair and within the laws of the game. PMID- 16046354 TI - Heading in football. Part 3: effect of ball properties on head response. AB - OBJECTIVES: Head impacts from footballs are an essential part of the game but have been implicated in mild and acute neuropsychological impairment. Ball characteristics have been noted in literature to affect the impact response of the head; however, the biomechanics are not well understood. The present study determined whether ball mass, pressure, and construction characteristics help reduce head and neck can impact response. METHODS: Head responses under ball impact (6-7 m/s) were measured with a biofidelic numerical human model and controlled human subject trials (n = 3). Three ball masses and four ball pressures were investigated for frontal heading. Further, the effect of ball construction in wet/dry conditions was studied with the numerical model. The dynamic ball characteristics were determined experimentally. Head linear and angular accelerations were measured and compared with injury assessment functions comprising peak values and head impact power. Neck responses were assessed with the numerical model. RESULTS: Ball mass reductions up to 35% resulted in decreased head responses up to 23-35% for the numerical and subject trials. Similar decreases in neck axial and shear responses were observed. Ball pressure reductions of 50% resulted in head and neck response reductions up to 10-31% for the subject trials and numerical model. Head response reductions up to 15% were observed between different ball constructions. The wet condition generally resulted in greater head and neck responses of up to 20%. CONCLUSION: Ball mass, pressure, and construction can reduce the impact severity to the head and neck. It is foreseeable that the benefits can be extended to players of all ages and skill levels. PMID- 16046355 TI - Effectiveness of headgear in football. AB - OBJECTIVES: Commercial headgear is currently being used by football players of all ages and skill levels to provide protection from heading and direct impact. The clinical and biomechanical effectiveness of the headgear in attenuating these types of impact is not well defined or understood. This study was conducted to determine whether football headgear has an effect on head impact responses. METHODS: Controlled laboratory tests were conducted with a human volunteer and surrogate head/neck system. The impact attenuation of three commercial headgears during ball impact speeds of 6-30 m/s and in head to head contact with a closing speed of 2-5 m/s was quantified. The human subject, instrumented to measure linear and angular head accelerations, was exposed to low severity impacts during heading in the unprotected and protected states. High severity heading contact and head to head impacts were studied with a biofidelic surrogate headform instrumented to measure linear and angular head responses. Subject and surrogate responses were compared with published injury assessment functions associated with mild traumatic brain injury (MTBI). RESULTS: For ball impacts, none of the headgear provided attenuation over the full range of impact speeds. Head responses with or without headgear were not significantly different (p>0.05) and remained well below levels associated with MTBI. In head to head impact tests the headgear provided an overall 33% reduction in impact response. CONCLUSION: The football headgear models tested did not provide benefit during ball impact. This is probably because of the large amount of ball deformation relative to headband thickness. However, the headgear provided measurable benefit during head to head impacts. PMID- 16046356 TI - Biomechanical investigation of head impacts in football. AB - OBJECTIVES: This study sought to measure the head accelerations induced from upper extremity to head and head to head impact during the game of football and relate this to the risk of mild traumatic brain injury using the Head Impact Power (HIP) index. Furthermore, measurement of upper neck forces and torques will indicate the potential for serious neck injury. More stringent rules or punitive sanctions may be warranted for intentional impact by the upper extremity or head during game play. METHODS: Game video of 62 cases of head impact (38% caused by the upper extremity and 30% by the head of the opposing player) was provided by F MARC. Video analysis revealed the typical impact configurations and representative impact speeds. Upper extremity impacts of elbow strike and lateral hand strike were re-enacted in the laboratory by five volunteer football players striking an instrumented Hybrid III pedestrian model crash test manikin. Head to head impacts were re-enacted using two instrumented test manikins. RESULTS: Elbow to head impacts (1.7-4.6 m/s) and lateral hand strikes (5.2-9.3 m/s) resulted in low risk of concussion (<5%) and severe neck injury (<5%). Head to head impacts (1.5-3.0 m/s) resulted in high concussion risk (up to 67%) but low risk of severe neck injury (<5%). CONCLUSION: The laboratory simulations suggest little risk of concussion based on head accelerations and maximum HIP. There is no biomechanical justification for harsher penalties in this regard. However, deliberate use of the head to impact another player's head poses a high risk of concussion, and justifies a harsher position by regulatory bodies. In either case the risk of serious neck injury is very low. PMID- 16046357 TI - Value of neuropsychological testing after head injuries in football. AB - This paper reviews the pros and cons of the traditional paper and pencil and the newer computerised neuropsychological tests in the management of sports concussion. The differences between diagnosing concussion on the field and neuropsychological assessment at follow up and decision making with regard to return to play are described. The authors also discuss the issues involved in interpreting the results of neuropsychological testing (comparison with population norms versus player's own baseline test results) and potential problems of such testing in football. Finally, suggested recommendations for neuropsychological testing in football are given. PMID- 16046358 TI - Reproducibility of computer based neuropsychological testing among Norwegian elite football players. AB - BACKGROUND: Head injuries account for 4-22% of all football injuries. The rate of brain injuries is difficult to assess, due to the problem of defining and grading concussion. Thus computerised testing programs for cognitive function have been developed. OBJECTIVE: To assess the reliability of a computerised neuropsychological test battery (CogSport) among Norwegian professional football players. METHODS: Norwegian professional football league players (90.3% participation) performed two consecutive baseline Cogsport tests before the 2004 season. CogSport consists of seven different subtasks: simple reaction time (SRT), choice reaction time (ChRT), congruent reaction time (CgRT), monitoring (MON), one-back (OBK), matching (Match) and learning (Learn). RESULTS: There was a small but significant improvement from repeated testing for the reaction time measurements of all seven subtasks (SRT: 0.7%, ChRT: 0.4%, CgRT: 1.2%, MON: 1.3%, OBK: 2.7%, Match: 2.0%, Learn: 1.1%). The coefficient of variation (CV) ranged from 1.0% to 2.7%; corresponding intraclass correlation coefficients ranged from 0.45 (0.34 to 0.55) to 0.79 (0.74 to 0.84). The standard deviation data showed higher CVs, ranging from 3.7% (Learn) to 14.2% (SRT). Thus, the variance decreased with increasing complexity of the task. The accuracy data displayed uniformly high CV (10.4-12.2) and corresponding low intraclass correlation coefficient (0.14 (0.01 to 0.26) to 0.31 (0.19 to 0.42)). CONCLUSION: The reproducibility for the mean reaction time measures was excellent, but less good for measures of accuracy and consistency. Consecutive testing revealed a slight learning effect from test 1 to test 2, and double baseline testing is recommended to minimise this effect. PMID- 16046359 TI - Effects of heading exposure and previous concussions on neuropsychological performance among Norwegian elite footballers. AB - BACKGROUND: Cross-sectional studies have indicated that neurocognitive performance may be impaired among football players. Heading the ball has been suggested as the cause, but recent reviews state that the reported deficits are more likely to be the result of head injuries. OBJECTIVE: To examine the association between previous concussions and heading exposure with performance on computer based neuropsychological tests among professional Norwegian football players. METHODS: Players in the Norwegian professional football league (Tippeligaen) performed two consecutive baseline neuropsychological tests (Cogsport) before the 2004 season (90.3% participation, n = 271) and completed a questionnaire assessing previous concussions, match heading exposure (self reported number of heading actions per match), player career, etc. Heading actions for 18 players observed in two to four matches were counted and correlated with their self-reported values. RESULTS: Neither match nor lifetime heading exposure was associated with neuropsychological test performance. Nineteen players scored below the 95% confidence interval for one or more subtasks, but they did not differ from the rest regarding the number of previous concussions or lifetime or match heading exposure. The number of previous concussions was positively associated with lifetime heading exposure (exponent (B) = 1.97(1.03-3.75), p = 0.039), but there was no relation between previous concussions and test performance. Self-reported number of headings correlated well with the observed values (Spearman's rho = 0.77, p < 0.001). CONCLUSION: Computerised neuropsychological testing revealed no evidence of neuropsychological impairment due to heading exposure or previous concussions in a cohort of Norwegian professional football players. PMID- 16046361 TI - Depressive symptoms are associated with increased systemic vascular resistance to stress. AB - OBJECTIVE: The deleterious effects of major depressive disorder on cardiovascular (CV) functioning are well known. However, the etiologic mechanisms underlying this association are incompletely understood. In the current study, subjects with varying degrees of depressive symptoms performed a stress task while CV reactivity was measured. We hypothesized that high levels of depressive symptoms would be associated with altered CV reactivity. METHODS: Ninety-one healthy volunteer subjects performed reactivity testing while measures of impedance cardiography and autonomic nervous system function were obtained. Subjects completed the Center for Epidemiological Studies Depression Scale (CES-D) and were categorized into either the high depressive (i.e., CES-D > or =16) or low depressive (i.e., CES-D <16) symptoms group. RESULTS: Task performance was associated with increases in systemic vascular resistance (SVR) (p = .001), mean arterial pressure (p = .001), and heart rate (p = .005), and decreases in cardiac output (p = .001), heather index (p = .001), and stroke volume (p = .05). After controlling for screening mean arterial pressure, an interaction effect of stress by mood group on SVR (p = .01) was observed; subjects with high amounts of depressive symptoms manifested significantly greater SVR at baseline and in response to a stressor task than did subjects with low amounts of depressive symptoms. CONCLUSIONS: These results suggest a mechanism that may partially explain the increased CV morbidity associated with depressive symptoms. In future studies, it may be useful to examine if treatment of depressive symptoms alters CV reactivity. PMID- 16046362 TI - Cardiovascular stress responses in young adulthood associated with family-of origin relationship experiences. AB - OBJECTIVE: The impact of relationships within the family-of-origin on the development of physiological stress responses has significant consequences for long-term vulnerability to stress-related illness. METHODS: The current study evaluated systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate (HR) responses to a challenge task in 150 young adults from families characterized by parental loss, divorce, or intact marriages. RESULTS: Overall, higher-quality family relationships were associated with stronger recovery of SBP. For DBP and HR, interactions were found in which higher-quality family relationships were associated with stronger recovery in the loss group relative to the divorce and intact groups. Good support was found for a mediational model outlining self-regulatory abilities as a pathway linking family relationships to SBP reactivity and recovery. CONCLUSIONS: Findings provide further evidence that family-of-origin relationship experiences can affect cardiovascular responses to later-life stress. PMID- 16046363 TI - Psychological distress as a predictor of CHD events in men: the effect of persistence and components of risk. AB - OBJECTIVE: This paper examines the role of psychological distress in the etiology of coronary heart disease (CHD), with particular reference to the persistence of distress symptoms, the contribution that undetected CHD at baseline makes to the observed associations and to the effect of separate components of psychological distress. METHOD: 5449 men in an occupational cohort (79% of the total), with at least two prior measurements of the General Health Questionnaire (GHQ-30), were followed for CHD events (including CHD death, nonfatal myocardial infarction (MI), and angina) for (mean) 6.8 years. Psychological distress was measured using the GHQ-30, and general/anxiety, depression and sleep subscales were created based on a principal components analysis. RESULTS: Psychological distress increased the risk of CHD events, with the risk highest in men with recent onset of distress. Age-adjusted hazard ratios were 1.48 (1.03-2.13) for persistent and 1.77 (1.13-2.78) for new distress. Angina events accounted for much of the observed associations. This increased risk was independent of conventional CHD risk factors, markers of underlying CHD, or measures of reporting bias, and it was related to anxiety items and sleep disturbance rather than depressive symptoms. CONCLUSIONS: Psychological distress increases the risk of a future diagnosis of angina in men. This risk is not accounted for by the presence of underlying CHD. These results highlight the importance of identifying both the role of underlying atherosclerosis in the pathway linking distress to heart disease and the timing of action of the components of psychological distress. PMID- 16046364 TI - Effects of partner support on resting oxytocin, cortisol, norepinephrine, and blood pressure before and after warm partner contact. AB - OBJECTIVE: We examined whether the magnitude of plasma oxytocin (OT), norepinephrine (NE), cortisol, and blood pressure (BP) responses before and after a brief episode of warm contact (WC) with the spouse/partner may be related to the strength of perceived partner support. METHODS: Subjects were 38 cohabiting couples (38 men, 38 women) aged 20 to 49 years. All underwent 10 minutes of resting baseline alone, 10 minutes of WC together with their partner, and 10 minutes of postcontact rest alone. RESULTS: Greater partner support (based on self-report) was related to higher plasma oxytocin in men and women across the protocol before and after WC. In women, higher partner support was correlated with lower systolic blood pressure (SBP) during solitary rest after WC but not before. Also, higher OT in women was linked to lower BP at baseline and to lower NE at all 4 measurements. CONCLUSION: Greater partner support is linked to higher OT for both men and women; however, the importance of OT and its potentially cardioprotective effects on sympathetic activity and BP may be greater for women. PMID- 16046365 TI - The effect of a telephone counseling intervention on self-rated health of cardiac patients. AB - OBJECTIVE: The objective of this study was to evaluate the effectiveness of a telephone-based intervention on psychological distress among patients with cardiac illness. METHODS: We recruited hospitalized patients surviving an acute coronary syndrome with scores on the Hospital and Anxiety Depression Scale (HADS) indicating mild to severe depression and/or anxiety at 1 month postdischarge. Recruited patients were randomized into either an intervention or control group. Intervention patients received up to six 30-minute telephone-counseling sessions focused on identifying cardiac-related fears. Control patients received usual care. For both groups, we collected patients' responses to the HADS and to the Global Improvement (CGI-I) subscale of the Clinical Global Impressions (CGI) Scale at baseline and at 2, 3, and 6 months postbaseline using Interactive Voice Recognition (IVR) technologies. We used mixed-effects analysis to estimate patients' changes in CGI-I measures over the three time points of data collection postbaseline. RESULTS: We enrolled 100 patients, and complete CGI-I measures were collected for 79 study patients. The mean age was 60 years (standard deviation = 10), and 67% of the patients were male. A mixed-effects analysis confirmed that patients in the intervention group had significantly greater improvements in self rated health (SRH) between baseline and month 3 than the control group (p = .01). Between month 3 and month 6, no significant differences in SRH improvements were observed between the control and intervention groups. CONCLUSIONS: Study patients reported greater SRH improvement resulting from the telephone-based intervention compared with control subjects. Future research should include additional outcome measures to determine the effect of changes in SRH on patients with comorbid physical and emotional disorders. PMID- 16046366 TI - Hostility scores are associated with increased risk of cardiovascular events in women undergoing coronary angiography: a report from the NHLBI-Sponsored WISE Study. AB - OBJECTIVE: To evaluate hostility-related personality traits assessed by the Cook Medley Hostility Inventory and to relate these to the occurrence of adverse cardiac events in women with suspected myocardial ischemia. METHODS: The cohort included 506 women with suspected coronary artery disease from the National Heart, Lung, and Blood Institute-sponsored Women's Ischemia Syndrome Evaluation (WISE) study. We examined individual components of the Cook Medley Hostility Score (CMHS) measuring cynicism, hostile affect, and aggressive responding, and a total CMHS (sum of these three) and associations with adverse events (defined as hospitalization for angina, nonfatal myocardial infarction, stroke, congestive heart failure (CHF) other vascular events and death) during 3 to 6 years follow up using Cox proportional hazard modeling. RESULTS: Women with adverse events had higher total CMHS (10.6 +/- 5.5) than women without any of these events (9.2 +/- 5.1) p = .02. They also had poorer survival by Kaplan-Meier analysis (log-rank p < .05). Unadjusted Cox models showed that the individual scores of cynicism and aggressive responding and the total CMHS were associated with more adverse events (all p < .05). Women with total CMHS above the median had a 35% increase risk of an adverse event in comparison to women with lower scores. In a risk-adjusted Cox model, the hazard ratio for an adverse event was 1.5 (p = .03) for women with total CMHS above the median. CONCLUSION: In this cohort of women with suspected myocardial ischemia, higher Cook Medley scores reflecting cynicism, hostile affect, and aggressive responding were associated with poorer 3 to 6 year event free survival and a higher risk of adverse events. After adjusting for risk factors and CAD, the association with risk for adverse events increased. PMID- 16046367 TI - A greater reduction in high-frequency heart rate variability to a psychological stressor is associated with subclinical coronary and aortic calcification in postmenopausal women. AB - OBJECTIVE: Reduced cardiac parasympathetic activity, as indicated by a reduced level of clinic or ambulatory high-frequency heart rate variability (HF-HRV), is associated with an increased risk for atherosclerosis and coronary artery disease. We tested whether the reduction in HF-HRV to a psychological stressor relative to a baseline level is also associated with subclinical coronary or aortic atherosclerosis, as assessed by calcification in these vascular regions. METHOD: Spectral estimates of 0.15 to 0.40 Hz HF-HRV were obtained from 94 postmenopausal women (61-69 years) who engaged in a 3-minute speech-preparation stressor after a 6-minute resting baseline. A median of 282 days later, electron beam tomography (EBT) was used to measure the extent of coronary and aortic calcification. RESULTS: In univariate analyses, a greater reduction in HF-HRV from baseline to speech preparation was associated with having more extensive calcification in the coronary arteries (rho = -0.29, p = .03) and in the aorta (rho = -0.22, p = .06). In multivariate analyses that controlled for age, education level, smoking status, hormone therapy use, fasting glucose, high density lipoproteins, baseline HF-HRV, and the stressor-induced change in respiration rate, a greater stressor-induced reduction in HF-HRV was associated with more calcification in the coronary arteries (B = -1.21, p < .05), and it was marginally associated with more calcification in the aorta (B = -0.92, p = .09). CONCLUSION: In postmenopausal women, a greater reduction in cardiac parasympathetic activity to a psychological stressor from baseline may be an independent correlate of subclinical atherosclerosis, particularly in the coronary arteries. PMID- 16046368 TI - Depressive symptoms and carotid artery intima-media thickness in young adults: the Cardiovascular Risk in Young Finns Study. AB - OBJECTIVE: Depression and coronary heart disease are often comorbid conditions, but the mechanism behind this link is largely unknown. We tested the hypothesis that a high level of depressive symptoms in healthy young adults would be related to more prevalent preclinical atherosclerosis. METHODS: We studied the association between depressive symptoms and carotid atherosclerosis in 1126 young adults (410 men and 716 women) as part of the ongoing population-based Cardiovascular Risk in Young Finns Study. The participants responded to a revised version of Beck's Depression Inventory in 1992, 1997, and 2001. Carotid atherosclerosis was assessed by measuring the thickness of the common carotid artery intima-media complex with ultrasound in 2001. Cardiovascular risk factors were measured in childhood/adolescence (1980) and in adulthood (2001). RESULTS: In men, high scorers of depressive symptoms in 2001 had higher carotid artery intima-media thickness (0.63 mm) compared with those with low or moderate scores on depressive symptoms (0.57 mm). This relationship (B = 0.08, F[1, 405] = 9.24, p = .003) persisted after adjustment for age and cardiovascular risk factors in adolescence and adulthood. Depression scores in 1992 and 1997 were not predictive of intima-media thickness. In women, no association was found between depressive symptoms and intima-media thickness. CONCLUSIONS: Depressive symptoms during early adulthood seem to be associated with higher levels of carotid intima-media thickness in men, but not in women. PMID- 16046369 TI - Mental disorders and revascularization procedures in a commercially insured sample. AB - OBJECTIVE: The objective of this study was to determine if receipt of revascularization was similar among commercially insured adults with mental disorders compared with people without mental disorders. METHODS: This was a retrospective analysis of a 100% sample of Blue Cross/Blue Shield of Iowa administrative claims data, 1996 to 2001. Logistic regression was used to calculate unadjusted and adjusted odds ratios (OR) for receipt of angioplasty (PTCA) and bypass graft surgery (CABG) within 30 days of discharge. RESULTS: A total of 3368 adults, aged 18 to 64 years, were hospitalized for myocardial infarction (MI) and 40% (n = 1342) had a mental disorder. Subjects with mental disorders were more likely to be younger, female, urban residents, and to have increased cardiovascular and medical comorbidity. They were similarly likely as subjects without mental disorders to have received PTCA (OR, 1.10; 95% confidence interval [CI], 0.95-1.29) and CABG (OR, 0.89; 95% CI, 0.71-1.11) in analyses adjusted for demographic and clinical characteristics. Revascularization rates did not differ by mental disorder type, with few exceptions. CONCLUSIONS: Receipt of revascularization was similar for patients with and without mental disorders. Our results may differ from previous findings as a result of the younger population studied and increased comorbidity in people with mental disorders, which may have resulted in a contraindication for surgical intervention. Conversely, the increased burden of comorbidity could suggest that these patients should have received PTCA at higher rates because of the better prognosis associated with revascularization as compared with medical management. Prospective analyses with review of clinical data and behavioral risk factors are necessary to determine why some patients with mental illness may be less likely to receive cardiac interventions. PMID- 16046370 TI - Relationship between work stress and body mass index among 45,810 female and male employees. AB - OBJECTIVE: The proportion of overweight and obese people has grown rapidly, and obesity has now been widely recognized as an important public health problem. At the same time, stress has increased in working life. The 2 problems could be connected if work stress promotes unhealthy eating habits and sedentary behavior and thereby contributes to weight gain. This study explored the association between work stress and body mass index (BMI; kg/m2). METHODS: We used cross sectional questionnaire data obtained from 45,810 female and male employees participating in the ongoing Finnish Public Sector Cohort Study. We constructed individual-level scores, as well as occupational- and organizational-level aggregated scores for work stress, as indicated by the demand/control model and the effort-reward imbalance model. Linear regression analyses were stratified by sex and socioeconomic status (SES) and adjusted for age, marital status, job contract, smoking, alcohol consumption, physical activity, and negative affectivity. RESULTS: The results with the aggregated scores showed that lower job control, higher job strain, and higher effort-reward imbalance were associated with a higher BMI. In men, lower job demands were also associated with a higher BMI. These associations were not accounted for by SES, although an additional adjustment for SES attenuated the associations. The results obtained with the individual-level scores were in the same direction, but the relationships were weaker than those obtained with the aggregated scores. CONCLUSIONS: This study shows a weak association between work stress and BMI. PMID- 16046371 TI - Cognitive and affective determinants of decisions to attend a group psychosocial support program for women with breast cancer. AB - OBJECTIVE: This prospective study assesses the roles of illness beliefs, emotion regulation factors, and sociodemographic characteristics in decisions to participate in a group support program for women recently diagnosed with breast cancer. METHOD: Women recruited during clinic visits 2 to 4 weeks after diagnosis completed measures of affective and cognitive factors identified by Leventhal's Common-Sense Model of illness self-regulation: cancer-related distress, avoidance tendencies, beliefs that the breast cancer was caused by stress and altered immunity, and personal control beliefs. Measures of general anxiety and depression, social support, and demographic characteristics were also completed; prognostic status information was obtained from medical records. All women were encouraged to participate in a free, 12-week program offering coping skills training and group support. Participation was recorded by program staff. RESULTS: Of the 110 women, 54 (49%) participated in the group support program and 56 (51%) did not. Logistic regression analyses revealed that participation was predicted by stronger beliefs that the cancer was caused by altered immunity, higher cancer related distress, lower avoidance tendencies, and younger age. CONCLUSIONS: Participation in the group psychosocial support program appeared to be guided by cognitive and affective factors identified by the Common-Sense Model. Psychosocial support programs and informational materials promoting their use may attract more participants if they are tailored to focus on resolving cancer related distress rather than on general anxiety or depression, appeal to those with high avoidance tendencies, address the role of immune function in cancer progression, and meet the needs of older participants. PMID- 16046372 TI - Central cholecystokinin activity in irritable bowel syndrome, panic disorder, and healthy controls. AB - OBJECTIVE: Irritable bowel syndrome (IBS) and panic disorder (PD) coexist with a high frequency. However, the nature of this relationship remains obscure. We have proposed that PD and IBS may share a common dysfunction of the central cholecystokinin (CCK) system. To test this hypothesis, we assessed whether the enhanced panicogenic response to CCK-tetrapeptide (CCK-4) observed in PD is also present in IBS. METHODS: Eight psychiatrically healthy IBS patients, 8 PD patients with no history of IBS, and 12 normal controls received a bolus injection of CCK-4 and placebo on two separate days in a double-blind, randomized fashion. RESULTS: Consistent with previous findings, panicogenic sensitivity to CCK-4 was enhanced in PD patients relative to controls. In contrast, IBS patients exhibited a response that was comparable to controls. Interestingly, CCK-4 induced nausea and abdominal distress were decreased in IBS patients relative to the other groups. No diagnostic difference was noted for cardiovascular response to CCK-4. CONCLUSION: These data indicate that IBS patients with no lifetime psychiatric history do not share the CCK-2 receptor dysfunction implicated in the pathophysiology of PD and that this dysfunction may not be a common mechanism for both CNS and enteric nervous system disorders. Nevertheless, the results suggest that a dysfunction of the CCK system may be involved in the pathophysiology of some enteric symptoms associated with IBS. PMID- 16046373 TI - Specific effects of depression, panic, and somatic symptoms on illness behavior. AB - OBJECTIVE: In terms of restricted financial resources, the contribution of mental disorders to health care use and illness behavior is highly relevant. However, the specific contributions of panic disorder, depression, and somatic complaints to illness behavior in unselected samples is unclear. METHODS: A representative sample of 2507 inhabitants of Germany was selected and grouped into people with panic disorder (30), somatic syndrome (102), major depression (24), depressive syndrome (77), and controls (2269). Assessment instruments were the complete version of the Patient Health Questionnaire (PHQ), the Scale for the Assessment of Illness Behavior (SAIB) as well as measures of health care use. RESULTS: Although all clinical groups showed increased illness behavior, their impact was independent and specific. Subjects fulfilling the criteria of panic disorder showed the highest scores for illness behavior and health care use. People with mental disorders showed a greater increase in visits to general practitioners and medical specialists than to psychiatrists or psychologists. Regression analysis revealed that somatic complaints and depression have independent associations with illness behavior and health care use. CONCLUSIONS: Different mental and psychosomatic disorders contribute independently to health care use and other aspects of illness behavior. PMID- 16046374 TI - Lung function and cognitive ability in a longitudinal birth cohort study. AB - OBJECTIVE: The objective of this study was to examine the association between forced expiratory volume in 1 second (FEV1) and cognitive ability in midlife in the normal population. METHODS: Multiple regression was used to test associations between FEV1 and cognitive function in 1778 men and women in the MRC National Survey of Health and Development, also known as the British 1946 birth cohort. Analyses were adjusted for sex, body size (birth weight, adult height, weight, and chest circumference), socioeconomic status, lifetime smoking, and a range of health indicators, including early respiratory vulnerability (infant lower respiratory infection, childhood asthma, and exposure to atmospheric pollution). RESULTS: FEV1 at 43 years was associated with slower psychomotor speed (peg placement) at the same age and with slower decline in psychomotor speed (letter search speed) from 43 to 53 years, independently of the previously mentioned potential confounders. These independent associations were not observed, however, for adult verbal ability, verbal memory, or rate of decline in memory, which were significantly explained by socioeconomic status and adolescent cognitive ability. In a subsequent analysis, adolescent cognition was positively associated with FEV1, although not with rate of decline in FEV1 from 43 to 53 years, again independently of the previously mentioned confounders. CONCLUSIONS: Cognitive function and FEV1 are positively associated across the life course. One possible explanation lies in the parallel action of endocrine, autonomic, and motor control systems on respiration and higher mental function. Because respiration and mental function are both associated with functional capacity and survival, this is a matter of potential clinical significance. PMID- 16046375 TI - Do depressive symptoms predict declines in physical performance in an elderly, biracial population? AB - OBJECTIVE: We investigated whether depressive symptoms, assessed by the 10-item Center for Epidemiological Studies Depression Scale (CES-D), predicted change in physical function in elderly adults. METHODS: Participants were from a biracial, population-based sample of adults aged 65 and older (N: 4069; 61% black; 61% female). Physical function was assessed as a summary performance measure of tandem stand, measured walk, and repeated chair stand (mean [standard deviation], 10.3 [3.5]; range, 0-15), commonly used measures of overall physical health in older adults. Generalized estimating equation models estimated physical function across 3 assessments over 5.4 years of follow up as a function of CES-D scores at baseline. RESULTS: Adjusting for age, sex, race, and education, each 1-point higher CES-D score was associated with a 0.34-point lower absolute level of physical performance (p < .0001), but there was no evidence of a CES-D by time interaction (p = .84), indicating that depressive symptoms at baseline were not associated with greater physical performance decline over time. In secondary analyses, with CES-D scores modeled in 4 categories, overall physical performance showed a graded, inverse association across CES-D categories (p's < .0001). However, we observed no threshold effect for depressive symptoms in relation to change in physical performance. Compared with the referent group (CES-D = 0), the 2 middle CES-D categories (CES-D = 1 or 2-3) evidenced some decline in physical performance over time, but the highest CES-D group (CES-D > or =4) showed no significant physical decline over time (p = .89). CONCLUSION: We observed a strong cross-sectional association between depressive symptoms and overall physical performance. Physical function declined over time, yet depressive symptoms did not consistently contribute to greater decline over an average of 5.4 years of follow up among older adults. Findings highlight the importance of longitudinal models in understanding the relation between depressive symptomatology and physical health. PMID- 16046376 TI - Colon motility during a panic attack. AB - OBJECTIVE: To document the temporal relationship between a panic attack and high amplitude propagating contractions. METHODS: Colon manometry was used to discriminate between functional defecation problems and colon neuromuscular disease. By chance, the patent developed a panic attack during the test session. RESULTS: Coincident with the panic attack, there was a continuous series of high amplitude propagating contractions. There were 15 high amplitude propagating contractions over 45 minutes, initially at a rate of 4 per 10 minutes, gradually slowing to 1.5 per 10 minutes. CONCLUSIONS: These data may explain the cause for gastrointestinal distress and diarrhea in some patients with panic attacks. PMID- 16046377 TI - The relationships among risk recognition, autonomic and self-reported arousal, and posttraumatic stress symptomatology in acknowledged and unacknowledged victims of sexual assault. AB - OBJECTIVE: This study examined differences among acknowledged sexual assault victims, unacknowledged sexual assault victims, and nonvictims in risk recognition and its psychophysiological correlates. METHODS: Acknowledged and unacknowledged victims of sexual assault and nonvictims (n = 97) listened to a hypothetical date rape interaction and were asked to indicate the point at which the man had become sexually inappropriate. Self-report and physiological measures of emotional responding and a measure of risk recognition were used to evaluate both between- and within-subjects' differences. RESULTS: Relative to nonvictims and acknowledged victims, unacknowledged victims of sexual assault took significantly longer to recognize risk. Acknowledged victims displayed decreased heart rate activity to a portion of the hypothetical interaction, but self reported greater arousal in response to the interaction and greater posttraumatic stress arousal symptomatology relative to nonvictims only. Posttraumatic stress symptomatology was related to self-reported emotional reactivity and response latency. CONCLUSIONS: The findings suggest that acknowledged and unacknowledged victims exhibit specific autonomic and behavioral response patterns that may perpetuate the cycle of traumatization. PMID- 16046378 TI - Prenatal stress alters cytokine levels in a manner that may endanger human pregnancy. AB - OBJECTIVE: Recent data suggest that prenatal stress negatively affects pregnancy and infant outcome. Existing studies implicate dysregulation of the immune and endocrine systems in stress-related increases in premature labor and poor birth outcome, but no published studies have directly addressed the relationships among these variables during pregnancy. We sought to test the hypothesis that high levels of psychosocial stress and low levels of social support during pregnancy alter maternal cytokine profiles in a manner that contributes to poor birth outcomes. METHODS: Psychosocial stress and social support were measured in 24 women with overtly normal pregnancies once during each trimester of pregnancy. Levels of interleukin-10 (IL-10), IL-6, and tumor necrosis factor-alpha (TNF alpha) were assessed concurrently with stress and support measurements. RESULTS: High social support was associated with low stress scores. Elevated stress scores were positively correlated with higher levels of the proinflammatory cytokines IL 6 and TNF-alpha, and with low levels of the antiinflammatory cytokine IL-10. CONCLUSIONS: These findings provide initial support for our hypothesis that stress-related neural immune interactions may contribute to pregnancy complications and poor outcome, but require further study to determine the mechanism and significance of these effects. PMID- 16046379 TI - Personality factors as determinants of depression in postpartum women: a prospective 1-year follow-up study. AB - OBJECTIVE: Personality has been associated with clinical depression in general. However, few studies have investigated personality in relation to postpartum depression, and these studies reported inconclusive findings. Therefore, the present study focused on neuroticism and introversion in the prediction of postpartum depression. METHOD: In a population-based prospective study, women were screened during mid-pregnancy on standard risk factors for depression. In a group of randomly selected women (n = 277), neuroticism and introversion were measured at 32 weeks gestation. Clinical depression (Research Diagnostic Criteria) and depressive symptoms (Edinburgh Postnatal Depression Scale) were measured at 32 weeks gestation and at 3, 6, and 12 months postpartum. RESULTS: High neuroticism was associated with an increased risk of clinical depression and depressive symptoms during the postpartum period. The combination of high neuroticism and high introversion was the only independent predictor of clinical depression across the first year postpartum (odds ratios: 3.08, 4.64, and 6.83 at 3, 6, and 12 months postpartum, respectively, p < .05-.01), even when controlling for clinical depression during pregnancy. History of depression was the only other independent predictor during the early but not during the late postpartum. Inclusion of personality not only significantly improved the detection of women at increased depression risk but also the identification of women with an extremely low depression risk. CONCLUSIONS: Personality may be an important and stable determinant of postpartum depression. The combination of high neuroticism and high introversion considerably improved the risk estimates for clinical depression across the first year postpartum. PMID- 16046380 TI - Depressive symptoms predict norepinephrine response to a psychological stressor task in Alzheimer's caregivers. AB - OBJECTIVE: Increased depressive symptoms have been associated with increased basal plasma norepinephrine (NE), and may be associated with exaggerated NE response to stress. This exaggerated response may play a role in health consequences associated with caring for a loved-one with Alzheimer's disease. The current study examined the relations between the level of depressive symptoms in spousal caregivers and the physiological response to a psychological stress task. METHODS: Fifty-five spousal caregivers (mean age 73 +/- 8 years) completed the depression subscale of the Brief Symptom Inventory (BSI). Plasma NE levels were assessed before and immediately after a speech stressor conducted at the caregiver's home. Hierarchical linear regression was used to determine whether caregiver depressive symptoms significantly improved prediction of change in NE levels beyond other factors theoretically and empirically related to NE. RESULTS: Level of depressive symptoms significantly predicted post-stressor change in NE levels (p < .01), even when controlling for age, caregiver distress, presence of caregiver hypertension, and care recipient level of cognitive function. Higher levels of depressive symptoms were associated with a greater plasma NE response to the psychological stress task. CONCLUSIONS: Depressive symptoms may act to exaggerate NE response to the stress of caregiving, potentially leading to an allostatic load that might predispose caregivers to negative health consequences, including cardiovascular morbidity. PMID- 16046381 TI - Repeated thermal therapy diminishes appetite loss and subjective complaints in mildly depressed patients. AB - OBJECTIVE: We observed that repeated thermal therapy improved appetite loss and general well-being in patients with chronic heart failure. The purpose of this study is to clarify the effects of repeated thermal therapy in mildly depressed patients with appetite loss and subjective complaints. METHODS: Twenty-eight mildly depressed inpatients with general fatigue, appetite loss, and somatic and mental complaints were randomly assigned to thermal therapy group (n = 14) or nonthermal therapy group (n = 14). Patients in the thermal therapy group were treated with 60 degrees C far-infrared ray dry sauna for 15 minutes and were then kept at bed rest with a blanket for 30 minutes once a day, 5 days a week for a total of 20 sessions in 4 weeks. RESULTS: Four weeks after admission, somatic complaints, hunger, and relaxation scores significantly improved (p < .001, p < .0001, p < .0001, respectively) and mental complaints slightly improved (p = .054) in the thermal therapy group compared with the nonthermal therapy group. Furthermore, the plasma ghrelin concentrations and daily caloric intake in the thermal therapy group significantly increased compared with the nonthermal therapy group (p < .05). CONCLUSIONS: These findings suggest that repeated thermal therapy may be useful for mildly depressed patients with appetite loss and subjective complaints. PMID- 16046382 TI - Baroreflex sensitivity is reduced in depression. AB - OBJECTIVES: Depression is independently associated with increased cardiovascular morbidity and mortality, including sudden cardiac death, and this risk is observed even in patients who have been successfully treated for depression. Recent studies have emphasized the importance of impaired baroreceptor sensitivity (BRS) as a predisposing factor for sudden death in patients with manifest cardiac disease. Our objective was to test the hypothesis that BRS is impaired in subjects with depression in remission and with no other cardiac risk factors. METHODS: We measured BRS by the sequence method in 36 patients with treated recurrent depression, who were euthymic at the time of study and with no manifest cardiac disease or "conventional" cardiac risk factors, compared with 39 healthy controls. Exclusion criteria included manifest heart disease or any risk factor for IHD (smoking, hypertension, diabetes, hypercholesterolemia, or body mass index >30). Nine subjects were not on any medication, and 22 were taking antidepressants. None of the controls was taking any medication. RESULTS: BRS was significantly lower in patients than in controls (19.5 [1.78] versus 25.4 [1.69] ms/mm Hg, p = .017). Analysis of covariance, in which age, sex, cholesterol, and body mass index were included, also showed that depression was a significant (p = .027) predictor of BRS. There was no significant difference in BRS adjusted by age and sex between the subjects taking antidepressants compared with those on no medications (p = .40). CONCLUSIONS: These data indicate that BRS is impaired in otherwise healthy patients with depression and may contribute to their increased cardiac risk. PMID- 16046383 TI - Analysis of variance is easily misapplied in the analysis of randomized trials: a critique and discussion of alternative statistical approaches. AB - Analysis of variance (ANOVA) is a statistical method that is widely used in the psychosomatic literature to analyze the results of randomized trials, yet ANOVA does not provide an estimate for the difference between groups, the key variable of interest in a randomized trial. Although the use of ANOVA is frequently justified on the grounds that a trial incorporates more than two groups, the hypothesis tested by ANOVA for these trials--"Are all groups equivalent?"--is often scientifically uninteresting. Regression methods are not only applicable to trials with many groups, but can be designed to address specific questions arising from the study design. ANOVA is also frequently used for trials with repeated measures, but the consequent reporting of "group effects," "time effects," and "time-by-group interactions" is a distraction from statistics of clinical and scientific value. Given that ANOVA is easily misapplied in the analysis of randomized trials, alternative approaches such as regression methods should be considered in preference. PMID- 16046384 TI - Chronic bronchitis, cigarette smoking, and the subsequent onset of depression and anxiety: results from a prospective population-based cohort study. AB - OBJECTIVE: The authors used data from a prospective, population-based cohort study to examine: (a) whether the presence of chronic bronchitis predicts the subsequent onset of depression or anxiety, and (b) if the incidence of depressed or anxious cases was different for smokers compared with nonsmokers. MATERIALS AND METHODS: For studying the relation between chronic bronchitis and anxiety or depression, we used data from respectively 4468 and 4520 respondents. RESULTS: The number of incident anxious (19.1%, n = 17) and depressed (14.0%, n = 13) cases was highest in employees with chronic bronchitis compared with employees without respiratory complaints (4.3%, n = 189 and 3.3%, n = 145, respectively). The presence of chronic bronchitis was associated with a significant increase in anxious and depressed cases (odds ratio (OR) for anxiety = 5.09, 95% confidence interval (CI) 2.91, 8.89; OR for depression = 4.38, 95% CI 2.35, 8.16). The incidence of anxiety as well as depression was strongest in the smokers group (OR for anxiety = 8.94, 95% CI 4.08, 19.59; OR for depression = 7.56, 95% CI 3.37, 16.96). CONCLUSIONS: This prospective study shows significantly higher levels of anxiety as well as depression in employees with chronic bronchitis. Results also seem to indicate that smoking cigarettes modifies this association, resulting in an increased risk of depression and anxiety in employees with chronic bronchitis who smoke. PMID- 16046385 TI - End-tidal pCO2 in blood phobics during viewing of emotion- and disease-related films. AB - OBJECTIVE: Many patients with blood, injection, and injury (BII) phobia respond to specific stimuli with vasovagal dysregulation and fainting. However, little is known about the role of hyperventilation in the distress of these patients. Hyperventilation, defined by subnormal arterial pCO2 levels, induces anxiety and may promote the development of fainting. We studied end-tidal pCO2 in 12 patients with BII phobia and 14 nonanxious controls during presentation of emotional films. METHOD: Ten film clips were shown, two in each of 5 categories: pleasant, unpleasant, neutral, BII-related (surgery), and asthma-related (portraying labored breathing). For each subject, two subsets were created, each containing one clip from each category. For one subset, the instruction was simply to view the film, and for the other subset, to view the film while tensing the leg muscles. PCO2, heart rate, blood pressure, and leg electromyogram were recorded continuously during viewing, and self-report of symptoms and emotion was collected after each film. RESULTS: Patients reported the greatest anxiety and disgust during surgery films. PCO2 was relatively stable throughout all categories except surgery films, during which minima were below 30 mm Hg, indicating significant hypocapnia. Cardiovascular variables suggested biphasic patterns in two patients with BII phobia. These patients, together with one additional patient and one control who were close to fainting after or during one surgery film, also showed a marked fall in pCO2) Leg muscle tension raised heart rate and systolic blood pressure for all films, but was not related to near fainting or endurance in surgery film viewing. CONCLUSION: Hyperventilation is part of the fear response of patients with BII phobia, but was transitory in experimental fear induction using surgery films. Its role in real-life exposure and fainting deserves further study. PMID- 16046386 TI - Glucose tolerance predicts short-term refeeding outcome in females with anorexia nervosa. AB - OBJECTIVE: Little is known about biologic predictors of refeeding outcome in anorexia nervosa (AN). Because nutritional status mirrors glucose metabolism during an oral glucose tolerance test (OGTT) in AN, this study investigated whether pretreatment glucose response patterns during the OGTT might be associated with refeeding progress in patients with AN. METHODS: Sixty-four female patients with anorexia (33 restrictors and 31 binge/purgers) and 13 healthy control subjects underwent an OGTT before nutritional rehabilitation, including desensitization to fear of energy intake of 1000 to 1600 kcal/day. Patients were divided into flat-type responders, impaired glucose tolerance (IGT) type responders, and normal-type glucose responders. Daily energy intake, weekly weight gain, and the duration of desensitization period were evaluated until the 12th week. RESULTS: The patients with anorexia consisted of 20 flat-type, 21 IGT type, and 23 normal- type responders. Normal-type responders required a shorter time to complete the desensitization period than other responders (p = .003 for restrictors, p < .001 for binge/purgers). In terms of refeeding progress, significant group effects for daily energy intake and weekly weight gain were evident in restrictors (p = .006, p = .028, respectively) and binge/purgers (p < .001, p = .003, respectively); normal-type responders showed good refeeding progress compared with other responders in both AN subtypes. CONCLUSIONS: The present study found a close relationship between pretreatment glucose responses, therapeutic progress of desensitization to fear of energy intake, and refeeding progress in both AN subtypes. Our findings suggest that glucose tolerance may be a useful predictor of short-term refeeding outcome in this disorder. PMID- 16046387 TI - Lost between the cracks: pain patients denied inpatient treatment for illicit drug addiction. PMID- 16046388 TI - Placebo research and the spirit of informed consent. PMID- 16046389 TI - Leukotriene B4 receptor and the function of its helix 8. AB - More than 30 lipid ligands, which express their biological activities through cognate G-protein-coupled receptors (GPCRs), have been reported. Among them, leukotriene B(4) (LTB(4)) is a potent lipid mediator involved in host defense, inflammation, and the immune responses. Two GPCRs for LTB(4) (BLT1 and BLT2) have been cloned and analyzed. Recent studies using genetically engineered mice suggest that BLT1 plays an important role in several inflammatory diseases including ischemic reperfusion tissue injury, atherosclerosis, and bronchial asthma. BLT1 is also a good tool to study the molecular mechanism of GPCR activation and inactivation in vitro. In this brief review, we focus on the biological and biochemical properties of BLT1 with special attention to the putative helix 8 of the receptor. PMID- 16046390 TI - The conformation of a signal peptide bound by Escherichia coli preprotein translocase SecA. AB - To understand the structural nature of signal sequence recognition by the preprotein translocase SecA, we have characterized the interactions of a signal peptide corresponding to a LamB signal sequence (modified to enhance aqueous solubility) with SecA by NMR methods. One-dimensional NMR studies showed that the signal peptide binds SecA with a moderately fast exchange rate (Kd approximately 10(-5) m). The line-broadening effects observed from one-dimensional and two dimensional NMR spectra indicated that the binding mode does not equally immobilize all segments of this peptide. The positively charged arginine residues of the n-region and the hydrophobic residues of the h-region had less mobility than the polar residues of the c-region in the SecA-bound state, suggesting that this peptide has both electrostatic and hydrophobic interactions with the binding pocket of SecA. Transferred nuclear Overhauser experiments revealed that the h region and part of the c-region of the signal peptide form an alpha-helical conformation upon binding to SecA. One side of the hydrophobic core of the helical h-region appeared to be more strongly bound in the binding pocket, whereas the extreme C terminus of the peptide was not intimately involved. These results argue that the positive charges at the n-region and the hydrophobic helical h-region are the selective features for recognition of signal sequences by SecA and that the signal peptide-binding site on SecA is not fully buried within its structure. PMID- 16046391 TI - Transforming activity of the Rho family GTPase, Wrch-1, a Wnt-regulated Cdc42 homolog, is dependent on a novel carboxyl-terminal palmitoylation motif. AB - Wrch-1 is a Rho family GTPase that shares strong sequence and functional similarity with Cdc42. Like Cdc42, Wrch-1 can promote anchorage-independent growth transformation. We determined that activated Wrch-1 also promoted anchorage-dependent growth transformation of NIH 3T3 fibroblasts. Wrch-1 contains a distinct carboxyl-terminal extension not found in Cdc42, suggesting potential differences in subcellular location and function. Consistent with this, we found that Wrch-1 associated extensively with plasma membrane and endosomes, rather than with cytosol and perinuclear membranes like Cdc42. Like Cdc42, Wrch-1 terminates in a CAAX tetrapeptide (where C is cysteine, A is aliphatic amino acid, and X is any amino acid) motif (CCFV), suggesting that Wrch-1 may be prenylated similarly to Cdc42. Most surprisingly, unlike Cdc42, Wrch-1 did not incorporate isoprenoid moieties, and Wrch-1 membrane localization was not altered by inhibitors of protein prenylation. Instead, we showed that Wrch-1 is modified by the fatty acid palmitate, and pharmacologic inhibition of protein palmitoylation caused mislocalization of Wrch-1. Most interestingly, mutation of the second cysteine of the CCFV motif (CCFV > CSFV), but not the first, abrogated both Wrch-1 membrane localization and transformation. These results suggest that Wrch-1 membrane association, subcellular localization, and biological activity are mediated by a novel membrane-targeting mechanism distinct from that of Cdc42 and other isoprenylated Rho family GTPases. PMID- 16046392 TI - Domains and maturation processes that regulate the activity of ADAMTS-2, a metalloproteinase cleaving the aminopropeptide of fibrillar procollagens types I III and V. AB - Processing of fibrillar collagens is required to generate collagen monomers able to self-assemble into elongated and cylindrical collagen fibrils. ADAMTS-2 belongs to the "A disintegrin and metalloproteinase with thrombospondin type 1 motifs" (ADAMTS) family. It is responsible for most of the processing of the aminopropeptide of type I procollagen in the skin, and it also cleaves type II and type III procollagens. ADAMTS are complex secreted enzymes that are implicated in various physiological and pathological processes. Despite accumulating evidence indicating that their activity is regulated by ancillary domains, additional information is required for a better understanding of the specific function of each domain. We have generated 17 different recombinant forms of bovine ADAMTS-2 and characterized their processing, activity, and cleavage specificity. The results indicated the following: (i) activation of the ADAMTS-2 zymogen involves several cleavages, by proprotein convertases and C terminal processing, and generates at least seven distinct processed forms; (ii) the C-terminal domain negatively regulates enzyme activity, whereas two thrombospondin type 1 repeats are enhancer regulators; (iii) the 104-kDa form displays the highest aminoprocollagen peptidase activity on procollagen type I; (iv) ADAMTS-2 processes the aminopropeptide of alpha1 type V procollagen homotrimer at the end of the variable domain; and (v) the cleaved sequence (PA) is different from the previously described sites ((P/A)Q) for ADAMTS-2, redefining its cleavage specificity. This finding and the existence of multiple processed forms of ADAMTS-2 strongly suggest that ADAMTS-2 may be involved in function(s) other than processing of fibrillar procollagen types I-III. PMID- 16046393 TI - Differential membrane localization of ERas and Rheb, two Ras-related proteins involved in the phosphatidylinositol 3-kinase/mTOR pathway. AB - Two Ras-related proteins, ERas and Rheb, which are involved in the phosphatidylinositol 3-kinase pathway, display high GTP affinity and have atypical CAAX motifs. The factors governing the intracellular localization of ERas and Rheb are incompletely understood. In the current study, we show by confocal microscopy that ERas is localized to the plasma membrane, whereas Rheb is confined to the endomembranes. Membrane localization of the two proteins was abolished by mutation of the cysteine of the CAAX motif. Membrane targeting was also abolished by a farnesyltransferase inhibitor but not by a geranylgeranyltransferase inhibitor. In mouse fibroblasts deficient in either Rce1 (Ras converting enzyme 1) or Icmt (isoprenylcysteine carboxyl methyltransferase), ERas was mislocalized mainly to the Golgi apparatus, whereas Rheb showed diffuse localization. Mutation of cysteines in the hypervariable region of ERas prevented the plasma membrane localization of ERas, very strongly suggesting that palmitoylation of the cysteines is essential for membrane targeting. The hypervariable region of Rheb does not contain cysteines or polybasic residues, and when it was replaced with the hypervariable region of H Ras, Rheb displayed plasma membrane localization. These data indicate that ERas shares the same posttranslational modifications with H-Ras and N-Ras and is localized at the plasma membrane. Rheb also shares the same membrane-targeting pathway but because of the absence of palmitoylation is located on endomembranes. PMID- 16046394 TI - Contribution of nuclear factor of activated T cells c1 to the transcriptional control of immunoreceptor osteoclast-associated receptor but not triggering receptor expressed by myeloid cells-2 during osteoclastogenesis. AB - Bone homeostasis depends on the coordination of osteoclastic bone resorption and osteoblastic bone formation. Receptor activator of NF-kappaB ligand (RANKL) induces osteoclast differentiation through activating a transcriptional program mediated by the key transcription factor nuclear factor of activated T cells (NFAT) c1. Immunoreceptors, including osteoclast-associated receptor (OSCAR) and triggering receptor expressed by myeloid cells (TREM)-2, constitute the co stimulatory signals required for RANKL-mediated activation of calcium signaling, which leads to the activation of NFATc1. However, it remains unknown whether the expression of immunoreceptors are under the control of NFATc1. Here we demonstrate that the expression of OSCAR, but not that of TREM-2, is up-regulated during osteoclastogenesis and markedly suppressed by the calcineurin inhibitor FK506, suggesting that OSCAR is transcriptionally regulated by NFATc1. NFATc1 expression results in the activation of the OSCAR promoter, which was found to be further enhanced by co-expression of PU.1 and microphthalmia-associated transcription factor (MITF). We further provide evidence that NFATc1 specifically regulates OSCAR by chromatin immunoprecipitation assay and quantification of OSCAR and TREM-2 mRNA in NFATc1-/- cells. Thus, OSCAR but not TREM-2 is involved in the positive feedback loop of the immunoreceptor-NFATc1 pathway during osteoclastogenesis. Although several immunoreceptors have been identified as co stimulatory molecules for RANKL, the expression and function are differentially regulated. These mechanisms, possibly together with the delicate regulation of their ligands on osteoblasts, may provide the exquisite machinery for the modulation of osteoclastogenesis in the maintenance of bone homeostasis. PMID- 16046395 TI - Allelic expression imbalance of human mu opioid receptor (OPRM1) caused by variant A118G. AB - As a primary target for opioid drugs and peptides, the mu opioid receptor (OPRM1) plays a key role in pain perception and addiction. Genetic variants of OPRM1 have been implicated in predisposition to drug addiction, in particular the single nucleotide polymorphism A118G, leading to an N40D substitution, with an allele frequency of 10-32%, and uncertain functions. We have measured allele-specific mRNA expression of OPRM1 in human autopsy brain tissues, using A118G as a marker. In 8 heterozygous samples measured, the A118 mRNA allele was 1.5-2.5-fold more abundant than the G118 allele. Transfection into Chinese hamster ovary cells of a cDNA representing only the coding region of OPRM1, carrying adenosine, guanosine, cytidine, and thymidine in position 118, resulted in 1.5-fold lower mRNA levels only for OPRM1-G118, and more than 10-fold lower OPRM1 protein levels, measured by Western blotting and receptor binding assay. After transfection and inhibition of transcription with actinomycin D, analysis of mRNA turnover failed to reveal differences in mRNA stability between A118 and G118 alleles, indicating a defect in transcription or mRNA maturation. These results indicate that OPRM1-G118 is a functional variant with deleterious effects on both mRNA and protein yield. Clarifying the functional relevance of polymorphisms associated with susceptibility to a complex disorder such as drug addiction provides a foundation for clinical association studies. PMID- 16046396 TI - The extracellular matrix and inflammation: fibromodulin activates the classical pathway of complement by directly binding C1q. AB - Components that propagate inflammation in joint disease may be derived from cartilage since the inflammation resolves after joint replacement. We found that the cartilage component fibromodulin has the ability to activate an inflammatory cascade, i.e. complement. Fibromodulin and immunoglobulins cause comparable deposition of C1q, C4b, and C3b from human serum. Using C1q and factor B deficient sera in combination with varying contents of metal ions, we established that fibromodulin activates both the classical and the alternative pathways of complement. Further studies revealed that fibromodulin binds directly to the globular heads of C1q, leading to activation of C1. However, deposition of the membrane attack complex and C5a release were lower in the presence of fibromodulin as compared with IgG. This can be explained by the fact that fibromodulin also binds complement inhibitor factor H. Factor H and C1q bind to non-overlapping sites on fibromodulin, but none of the interactions is mediated by the negatively charged keratan sulfate substituents of fibromodulin. C1q but not factor H binds to an N-terminal fragment of fibromodulin previously implicated to be affected in cartilage stimulated with the inflammatory cytokine interleukin 1. Taken together our observations indicate fibromodulin as one factor involved in the sustained inflammation of the joint. PMID- 16046397 TI - The non-gastric H,K-ATPase is oligomycin-sensitive and can function as an H+,NH4(+)-ATPase. AB - We used the baculovirus/Sf9 expression system to gain new information on the mechanistic properties of the rat non-gastric H,K-ATPase, an enzyme that is implicated in potassium homeostasis. The alpha2-subunit of this enzyme (HKalpha2) required a beta-subunit for ATPase activity thereby showing a clear preference for NaKbeta1 over NaKbeta3 and gastric HKbeta. NH4(+), K+, and Na+ maximally increased the activity of HKalpha2-NaKbeta1 to 24.0, 14.2, and 5.0 micromol P(i) x mg(-1) protein x h(-1), respectively. The enzyme was inhibited by relatively high concentrations of ouabain and SCH 28080, whereas it was potently inhibited by oligomycin. From the phosphorylation level in the presence of oligomycin and the maximal NH4(+)-stimulated ATPase activity, a turnover number of 20,000 min( 1) was determined. All three cations decreased the steady-state phosphorylation level and enhanced the dephosphorylation rate, disfavoring the hypothesis that Na+ can replace H+ as the activating cation. The potency with which vanadate inhibited the cation-activated enzyme decreased in the order K+ > NH4(+) > Na+, indicating that K+ is a stronger E2 promoter than NH4(+), whereas in the presence of Na+ the enzyme is in the E1 form. For K+ and NH4(+), the E2 to E1 conformational equilibrium correlated with their efficacy in the ATPase reaction, indicating that here the transition from E2 to E1 is rate-limiting. Conversely, the low maximal ATPase activity with Na+ is explained by a poor stimulatory effect on the dephosphorylation rate. These data show that NH4(+) can replace K+ with similar affinity but higher efficacy as an extracellular activating cation in rat nongastric H,K-ATPase. PMID- 16046398 TI - Potent mechanism-based inhibitors for matrix metalloproteinases. AB - Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that play important roles in physiological and pathological conditions. Both gelatinases (MMP-2 and -9) and membrane-type 1 MMP (MMP-14) are important targets for inhibition, since their roles in various diseases, including cancer, have been well established. We describe herein a set of mechanism-based inhibitors that show high selectivity to gelatinases and MMP-14 (inhibitor 3) and to only MMP-2 (inhibitors 5 and 7). These molecules bind to the active sites of these enzymes, initiating a slow binding profile for the onset of inhibition, which leads to covalent enzyme modification. The full kinetic analysis for the inhibitors is reported. These are nanomolar inhibitors (Ki) for the formation of the noncovalent enzyme-inhibitor complexes. The onset of slow binding inhibition is rapid (k(on) of 10(2) to 10(4) M(-1) s(-1) and the reversal of the process is slow (k(off) of 10(-3) to 10(-4) s(-1)). However, with the onset of covalent chemistry with the best of these inhibitors (e.g. inhibitor 3), very little recovery of activity (<10%) was seen over 48 h of dialysis. We previously reported that broad spectrum MMP inhibitors like GM6001 enhance MT1-MMP-dependent activation of pro-MMP-2 in the presence of tissue inhibitor of metalloproteinases 2. Herein, we show that inhibitor 3, in contrast to GM6001, had no effect on pro MMP-2 activation by MT1-MMP. Furthermore, inhibitor 3 reduced tumor cell migration and invasion in vitro. These results show that these new inhibitors are promising candidates for selective inhibition of MMPs in animal models of relevant human diseases. PMID- 16046399 TI - Dodecameric structure of the small heat shock protein Acr1 from Mycobacterium tuberculosis. AB - Small heat shock proteins are a ubiquitous and diverse family of stress proteins that have in common an alpha-crystallin domain. Mycobacterium tuberculosis has two small heat shock proteins, Acr1 (alpha-crystallin-related protein 1, or Hsp16.3/16-kDa antigen) and Acr2 (HrpA), both of which are highly expressed under different stress conditions. Small heat shock proteins form large oligomeric assemblies and are commonly polydisperse. Nanoelectrospray mass spectrometry showed that Acr2 formed a range of oligomers composed of dimers and tetramers, whereas Acr1 was a dodecamer. Electron microscopy of Acr2 showed a variety of particle sizes. Using three-dimensional analysis of negative stain electron microscope images, we have shown that Acr1 forms a tetrahedral assembly with 12 polypeptide chains. The atomic structure of a related alpha-crystallin domain dimer was docked into the density to build a molecular structure of the dodecameric Acr1 complex. Along with the differential regulation of these two proteins, the differences in their quaternary structures demonstrated here supports their distinct functional roles. PMID- 16046400 TI - Human high density lipoproteins are platforms for the assembly of multi-component innate immune complexes. AB - Human innate immunity to non-pathogenic species of African trypanosomes is provided by human high density lipoprotein (HDL) particles. Here we show that native human HDLs containing haptoglobin-related protein (Hpr), apolipoprotein L I (apoL-I) and apolipoprotein A-I (apoA-I) are the principle antimicrobial molecules providing protection from trypanosome infection. Other HDL subclasses containing either apoA-I and apoL-I or apoA-I and Hpr have reduced trypanolytic activity, whereas HDL subclasses lacking apoL-I and Hpr are non-toxic to trypanosomes. Highly purified, lipid-free Hpr and apoL-I were both toxic to Trypanosoma brucei brucei but with specific activities at least 500-fold less than those of native HDLs, suggesting that association of these apolipoproteins within the HDL particle was necessary for optimal cytotoxicity. These studies show that HDLs can serve as platforms for the assembly of multiple synergistic proteins and that these assemblies may play a critical role in the evolution of primate-specific innate immunity to trypanosome infection. PMID- 16046401 TI - T-oligo treatment decreases constitutive and UVB-induced COX-2 levels through p53 and NFkappaB-dependent repression of the COX-2 promoter. AB - Chronically irradiated murine skin and UV light-induced squamous cell carcinomas overexpress the inducible isoform of cyclooxygenase (COX-2), and COX-2 inhibition reduces photocarcinogenesis in mice. We have reported previously that DNA oligonucleotides substantially homologous to the telomere 3'-overhang (T-oligos) induce DNA repair capacity and multiple other cancer prevention responses, in part through up-regulation and activation of p53. To determine whether T-oligos affect COX-2 expression, human newborn keratinocytes and fibroblasts were pretreated with T-oligos or diluent alone for 24 h, UV-irradiated, and processed for Western blotting. In both cell types, T-oligos transcriptionally down regulated base-line and UV light-induced COX-2 expression, coincident with p53 activation. In fibroblasts with wild type versus dominant negative p53 (p53(WT) versus p53(DN)), T-oligos decreased constitutive expression of a COX-2 reporter plasmid by >50%. We then examined NFkappaB, a known positive regulator of COX-2 transcription. In p53(WT) but not in p53(DN) fibroblasts and in human keratinocytes, T-oligos decreased readout of an NFkappaB promoter-driven reporter plasmid and decreased NFkappaB binding to DNA. After T-oligo treatment and subsequent UV irradiation, binding of the transcriptional co-activator protein p300 to NFkappaB was decreased, whereas binding of p300 to p53 was increased. Human skin explants provided with T-oligos had markedly decreased COX-2 immunostaining both at base-line and post-UV light, coincident with increased p53 immunostaining. We conclude that T-oligos transcriptionally down-regulate COX-2 expression in human skin via activation and up-regulation of p53, at least in part by inhibiting NFkappaB transcriptional activation. Decreased COX-2 expression may contribute to the observed ability of T-oligos to reduce photocarcinogenesis. PMID- 16046402 TI - Serglycin-deficient cytotoxic T lymphocytes display defective secretory granule maturation and granzyme B storage. AB - Cytotoxic T lymphocytes eliminate infected and tumor cells mainly by perforin/granzyme-induced apoptosis. Earlier studies suggested that serglycin proteoglycans form macromolecular complexes with granzymes and perforin in the cytotoxic granule. Serglycin-proteoglycans may also be involved in the delivery of the cytolytic machinery into target cells. We have developed a serglycin deficient mouse strain, and here we studied the importance of serglycin proteoglycans for various aspects of cytotoxic T lymphocyte function. 35SO4(2-) radiolabeling of serglycin-deficient cells demonstrated a dramatic reduction of incorporated label as compared with wild type cells, indicating that serglycin is by far the dominating proteoglycan species produced by the cytotoxic T lymphocyte. Moreover, lack of serglycin resulted in impaired ability of cytotoxic T lymphocytes to produce secretory granule of high electron density, although granule of lower electron density were produced both in wild type and serglycin deficient cells. The serglycin deficiency did not affect the mRNA expression for granzyme A, granzyme B, or perforin. However, the storage of granzyme B, but not granzyme A, Fas ligand, or perforin, was severely defective in serglycin deficient cells. Serglycin-deficient cells did not display defects in late cytotoxicity toward target cell lines. Taken together, these results point to a key role for serglycin in the storage of granzyme B and for secretory granule maturation but argue against a major role for serglycin in the apoptosis mediated by cytotoxic T lymphocytes. PMID- 16046403 TI - Human organic anion transporter hOAT1 forms homooligomers. AB - Human organic anion transporter hOAT1 belongs to a superfamily of organic anion transporters, which play critical roles in the body disposition of clinically important drugs, including anti-human immunodeficiency virus therapeutics, anti tumor drugs, antibiotics, anti-hypertensives, and anti-inflammatories. To gain insight into the regulation of hOAT1, detailed information on its structural assembly is essential. In the present study, we investigate the quaternary structure of hOAT1 using combined approaches of chemical cross-linking, gel filtration chromatography, co-immunoprecipitation, cell surface biotinylation, and metabolic labeling. Chemical cross-linking of intact membrane proteins from LLC-PK1 cells stably expressing hOAT1 converted quantitatively hOAT1 monomer to putative trimer and higher order of oligomer, indicating that hOAT1 is present in the membrane as multimeric complexes. When co-expressed in LLC-PK1 cells, FLAG tagged hOAT1 co-immunoprecipitated with myc-tagged hOAT1. The hOAT1 oligomer was also detected in gel filtration chromatography of total membranes from hOAT1 expressing LLC-PK1 cells. Cell surface biotinylation with membrane-impermeable reagents and metabolic labeling with [(35)S]methionine followed by immunoprecipitation showed that the oligomeric hOAT1 did not contain any other proteins. Taken together, this is the first study demonstrating that hOAT1 exists in the plasma membrane as a homooligomer, possibly trimer, and higher order of oligomer. PMID- 16046404 TI - Myelin basic protein-primed T cells of female but not male mice induce nitric oxide synthase and proinflammatory cytokines in microglia: implications for gender bias in multiple sclerosis. AB - Females are more susceptible than males to multiple sclerosis (MS). However, the underlying mechanism behind this gender difference is poorly understood. Because the presence of neuroantigen-primed T cells within the CNS is necessary for the development of MS, the present study was undertaken to investigate the activation of microglia by myelin basic protein (MBP)-primed T cells of male, female, and castrated male mice. Interestingly, MBP-primed T cells isolated from female and castrated male but not from male mice induced the expression of inducible nitric oxide synthase (iNOS) and proinflammatory cytokines (interleukin-1beta (IL 1beta), IL-1alpha, IL-6, and tumor necrosis factor-alpha) in microglia by cell cell contact. Again there was no apparent defect in male microglia, because MBP primed T cells isolated from female and castrated male but not male mice were capable of inducing the production of NO in male primary microglia. Inhibition of female T cell contact-mediated microglial expression of proinflammatory molecules by dominant-negative mutants of p65 and C/EBPbeta suggest that female MBP-primed T cells induce microglial expression of proinflammatory molecules through the activation of NF-kappaB and C/EBPbeta. Interestingly, MBP-primed T cells of male, female, and castrated male mice were able to induce microglial activation of NF kappaB. However, MBP-primed T cells of female and castrated male but not male mice induced microglial activation of C/EBPbeta. These studies suggest that microglial activation of C/EBPbeta but not NF-kappaB by T cell:microglial contact is a gender-specific event and that male MBP-primed T cells are not capable of inducing microglial expression of proinflammatory molecules due to their inability to induce the activation of C/EBPbeta in microglia. This novel gender sensitive activation of microglia by neuroantigen-primed T cell contact could be one of the mechanisms behind the female-loving nature of MS. PMID- 16046405 TI - Phosphorylation of phosphophoryn is crucial for its function as a mediator of biomineralization. AB - Phosphoproteins of the organic matrix of bone and dentin have been implicated as regulators of the nucleation and growth of the inorganic Ca-P crystals of vertebrate bones and teeth. One such protein identified in the dentin matrix is phosphophoryn (PP). It is highly acidic in nature because of a high content of aspartic acid and phosphate groups on serines. The 244-residue carboxyl-terminal domain of rat PP, predominantly containing the aspartic acid-serine repeats, has been cloned, and the corresponding protein has been expressed recombinantly in Escherichia coli. This portion of PP, named DMP2 (dentin matrix protein 2), is not phosphorylated by the bacteria and thus provided a means to study the function of the phosphate groups, the major post-translational modification of native PP. The recombinant DMP2 (rDMP2) possessed much lower calcium binding capacity than native PP. Small angle x-ray scattering experiments demonstrated that PP folds to a compact globular structure upon calcium binding, whereas rDMP2 maintained an unfolded structure. In vitro nucleation experiments showed that PP could nucleate plate-like apatite crystals in pseudophysiological buffer, whereas rDMP2 failed to mediate the transformation of amorphous calcium phosphate to apatite crystals under the same experimental conditions. Collagen binding experiments demonstrated that PP favors the formation of collagen aggregates, whereas in the presence of rDMP2 thin fibrils are formed. Overall these results suggested that the phosphate moieties in phosphophoryn are important for its function as a mediator of dentin biomineralization. PMID- 16046406 TI - A nine-transmembrane domain topology for presenilin 1. AB - Presenilin (PS) provides the catalytic core of the gamma-secretase complex. Gamma secretase activity leads to generation of the amyloid beta-peptide, a key event implicated in the pathogenesis of Alzheimer disease. PS has ten hydrophobic regions, which can all theoretically form membrane-spanning domains. Various topology models have been proposed, and the prevalent view holds that PS has an eight-transmembrane (TM) domain organization; however, the precise topology has not been unequivocally determined. Previous topological studies are based on non functional truncated variants of PS proteins fused to reporter domains, or immunocytochemical staining. In this study, we used a more subtle N-linked glycosylation scanning approach, which allowed us to assess the topology of functional PS1 molecules. Glycosylation acceptor sequences were introduced into full-length human PS1, and the results showed that the first hydrophilic loop is oriented toward the lumen of the endoplasmic reticulum, whereas the N terminus and large hydrophilic loop are in the cytosol. Although this is in accordance with most current models, our data unexpectedly revealed that the C terminus localized to the luminal side of the endoplasmic reticulum. Additional studies on the glycosylation pattern after TM domain deletions, combined with computer-based TM protein topology predictions and biotinylation assays of different PS1 mutants, led us to conclude that PS1 has nine TM domains and that the C terminus locates to the lumen/extracellular space. PMID- 16046407 TI - Essential constituents of the 3'-phosphoesterase domain of bacterial DNA ligase D, a nonhomologous end-joining enzyme. AB - DNA ligase D (LigD) catalyzes end-healing and end-sealing steps during nonhomologous end joining in bacteria. Pseudomonas aeruginosa LigD consists of a central ATP-dependent ligase domain fused to a C-terminal polymerase domain and an N-terminal 3'-phosphoesterase (PE) module. The PE domain catalyzes manganese dependent phosphodiesterase and phosphomonoesterase reactions at a duplex primer template with a short 3'-ribonucleotide tract. The phosphodiesterase, which cleaves a 3'-terminal diribonucleotide to yield a primer strand with a ribonucleoside 3'-PO4 terminus, requires the vicinal 2'-OH of the penultimate ribose. The phosphomonoesterase converts the terminal ribonucleoside 3'-PO4 to a 3'-OH. Here we show that the PE domain has a 3'-phosphatase activity on an all DNA primer-template, signifying that the phosphomonoesterase reaction does not depend on a 2'-OH. The distinctions between the phosphodiesterase and phosphomonoesterase activities are underscored by the results of alanine scanning, limited proteolysis, and deletion analysis, which show that the two reactions depend on overlapping but nonidentical ensembles of protein functional groups, including: (i) side chains essential for both ribonuclease and phosphatase activity (His-42, His-48, Asp-50, Arg-52, His-84, and Tyr-88); (ii) side chains important for 3'-phosphatase activity but not for 3' ribonucleoside removal (Arg-14, Asp-15, Glu-21, Gln-40, and Glu-82); and (iii) side chains required selectively for the 3'-ribonuclease (Lys-66 and Arg-76). These constellations of critical residues are unique to LigD-like proteins, which we propose comprise a new bifunctional phosphoesterase family. PMID- 16046408 TI - Ultrasonication-induced amyloid fibril formation of beta2-microglobulin. AB - To obtain insight into the mechanism of fibril formation, we examined the effects of ultrasonication, a strong agitator, on beta2-microglobulin (beta2-m), a protein responsible for dialysis-related amyloidosis. Upon sonication of an acid unfolded beta2-m solution at pH 2.5, thioflavin T fluorescence increased markedly after a lag time of 1-2 h with a simultaneous increase of light scattering. Atomic force microscopy images showed the formation of a large number of short fibrils 3 nm in diameter. When the sonication-induced fibrils were used as seeds in the next seeding experiment at pH 2.5, a rapid and intense formation of long fibrils 3 nm in diameter was observed demonstrating seed-dependent fibril growth. We then examined the effects of sonication on the native beta2-m at neutral pH, conditions under which amyloid deposits occur in patients. In the presence of 0.5 mm sodium dodecyl sulfate, a model compound of potential trigger and stabilizer of amyloid fibrils in patients, a marked increase of thioflavin T fluorescence was observed after 1 day of sonication at pH 7.0. The products of sonication caused the accelerated fibril formation at pH 7.0. Atomic force microscopy images showed that the fibrils formed at pH 7.0 have a diameter of more than 7 nm, thicker than those prepared at pH 2.5. These results indicate that ultrasonication is one form of agitation triggering the formation of amyloid fibrils of beta2-m, producing fibrils adapted to the respective pH. PMID- 16046409 TI - Giardia lamblia RNA cap guanine-N2 methyltransferase (Tgs2). AB - Tgs1 is the enzyme responsible for converting 7-methylguanosine RNA caps to the 2,2,7-trimethylguanosine cap structures of small nuclear and small nucleolar RNAs. Whereas budding yeast Saccharomyces cerevisiae and fission yeast Schizosaccharomyces pombe encode a single Tgs1 protein, the primitive eukaryote Giardia lamblia encodes two paralogs, Tgs1 and Tgs2. Here we show that purified Tgs2 is a monomeric enzyme that catalyzes methyl transfer from AdoMet (K(m) of 6 microm) to m(7)GDP (K(m) of 65 microm; k(cat) of 14 min(-1)) to form m(2,7)GDP. Tgs2 also methylates m(7)GTP (K(m) of 30 microm; k(cat) of 13 min(-1)) and m(7)GpppA (K(m) of 7 microm; k(cat)) of 14 min(-1) but is unreactive with GDP, GTP, GpppA, ATP, CTP, or UTP. We find that the conserved residues Asp-68, Glu-91, and Trp-143 are essential for Tgs2 methyltransferase activity in vitro. The m(2,7)GDP product formed by Tgs2 can be converted to m(2,2,7)GDP by S. pombe Tgs1 in the presence of excess AdoMet. However, Giardia Tgs2 itself is apparently unable to add a second methyl group at guanine-N2. This result implies that 2,2,7 trimethylguanosine caps in Giardia are either synthesized by Tgs1 alone or by the sequential action of Tgs2 and Tgs1. The specificity of Tgs2 raises the prospect that some Giardia mRNAs might contain dimethylguanosine caps. PMID- 16046410 TI - Expression of an uncleavable N-terminal RasGAP fragment in insulin-secreting cells increases their resistance toward apoptotic stimuli without affecting their glucose-induced insulin secretion. AB - Apoptosis of pancreatic beta cells is implicated in the onset of type 1 and type 2 diabetes. Consequently, strategies aimed at increasing the resistance of beta cells toward apoptosis could be beneficial in the treatment of diabetes. RasGAP, a regulator of Ras and Rho GTPases, is an atypical caspase substrate, since it inhibits, rather than favors, apoptosis when it is partially cleaved by caspase-3 at position 455. The antiapoptotic signal generated by the partial processing of RasGAP is mediated by the N-terminal fragment (fragment N) in a Ras phosphatidylinositol 3-kinase-Akt-dependent, but NF-kappaB-independent, manner. Further cleavage of fragment N at position 157 abrogates its antiapoptotic properties. Here we demonstrate that an uncleavable form of fragment N activates Akt, represses NF-kappaB activity, and protects the conditionally immortalized pancreatic insulinoma betaTC-tet cell line against various insults, including exposure to genotoxins, trophic support withdrawal, and incubation with inflammatory cytokines. Fragment N also induced Akt activity and protection against cytokine-induced apoptosis in primary pancreatic islet cells. Fragment N did not alter insulin cell content and insulin secretion in response to glucose. These data indicate that fragment N protects beta cells without affecting their function. The pathways regulated by fragment N are therefore promising targets for antidiabetogenic therapy. PMID- 16046411 TI - Diet-induced obesity in C57BL/6J mice causes increased renal lipid accumulation and glomerulosclerosis via a sterol regulatory element-binding protein-1c dependent pathway. AB - Obesity and metabolic syndrome are associated with glomerulosclerosis and proteinuria, but the mechanisms are not known. The purpose of this study was to determine if there is altered renal lipid metabolism and increased expression of sterol regulatory element-binding proteins (SREBPs) in a model of diet-induced obesity. C57BL/6J mice that were fed a high fat, 60 kcal % saturated (lard) fat diet (HFD) developed obesity, hyperglycemia, and hyperinsulinemia compared with those that were fed a low fat, 10 kcal % fat diet (LFD). In contrast, A/J mice were resistant when fed the same diet. C57BL/6J mice with HFD exhibited significantly higher levels of renal SREBP-1 and SREBP-2 expression than those mice with LFD, whereas in A/J mice there were no changes with the same treatment. The increases in SREBP-1 and SREBP-2 expression in C57BL/6J mice resulted in renal accumulation of triglyceride and cholesterol. There were also significant increases in the renal expression of plasminogen activator inhibitor-1 (PAI-1), vascular endothelial growth factor (VEGF), type IV collagen, and fibronectin, resulting in glomerulosclerosis and proteinuria. To determine a role for SREBPs per se in modulating renal lipid metabolism and glomerulosclerosis we performed studies in SREBP-1c(-/-) mice. In contrast to control mice, in the SREBP-1c(-/-) mice with HFD the accumulation of triglyceride was prevented, as well as the increases in PAI-1, VEGF, type IV collagen, and fibronectin expression. Our results therefore suggest that diet-induced obesity causes increased renal lipid accumulation and glomerulosclerosis in C57BL/6J mice via an SREBP-1c-dependent pathway. PMID- 16046412 TI - Cellular uptake of mammalian heparanase precursor involves low density lipoprotein receptor-related proteins, mannose 6-phosphate receptors, and heparan sulfate proteoglycans. AB - Mammalian heparanase, strongly implicated in the regulation of cell growth, migration, and differentiation, plays a crucial role in inflammation, angiogenesis, and metastasis. There is thus a clear need for understanding how heparanase activity is regulated. Cells can generate an active form of the enzyme from a larger inactive precursor protein by a process of secretion-recapture, internalization, and proteolytic processing in late endosomes/lysosomes. Cell surface heparan sulfate proteoglycans are the sole known components with a role in this trafficking of the heparanase precursor. Here, we provide evidence that heparan sulfate proteoglycans are not strictly required for this process. More importantly, by heparanase transfection, binding, and uptake experiments and by using a combination of specific inhibitors and receptor-defective cells, we have identified low density lipoprotein receptor-related proteins and mannose 6 phosphate receptors as key elements of the receptor system that mediates the capture of secreted heparanase precursor and its trafficking to the intracellular site of processing/activation. PMID- 16046413 TI - The G alpha(o/i)-coupled cannabinoid receptor-mediated neurite outgrowth involves Rap regulation of Src and Stat3. AB - The study of the signaling pathways regulating neurite outgrowth in culture is important because of their potential role in neuronal differentiation in vivo. We have previously shown that the G alpha(o/i)-coupled CB1 cannabinoid receptor (CB1R) activates Rap1 to induce neurite outgrowth. G alpha(o/i) also activates the Src-Stat3 pathway. Here, we studied the relationship between the G alpha(o/i) Rap1 and Src-Stat3 pathways and the role of these signaling pathways in CB1R mediated neurite outgrowth in Neuro-2A cells. The CB1 agonist HU-210 induced pertussis toxin-sensitive Src and Stat3 phosphorylation. Dominant negative (DN) mutants of Src and Stat3 blocked CB1R-induced neurite outgrowth. Constitutively active Rap 1B and Ral-activated Src and CB1R-induced Src phosphorylation was inhibited by Rap1-DN and Ral-DN, indicating that both Rap1 and Ral mediate downstream signaling from G alpha(o/i) for Src activation. Rap1-activated Ral and Ral-DN blocked Rap-induced Src phosphorylation. G alpha(o)-induced Stat3 activation was blocked by Ral-DN, whereas v-Src-induced Stat3 activation was not inhibited by Ral-DN, indicating that the CB1R, through G alpha(o), mediates the sequential activation of Rap1 to Ral to Src to Stat3 in Neuro-2A cells. Downstream of Src, the CB1R also activated Rac1 and JNK, which enhanced CBR1 mediated Stat3 activation. Rac-DN blocked CB1R-induced activation of JNK. Pharmacological inhibition of JNK blocked Src and CB1R activation of Stat3, indicating that Rac and JNK are also involved in CB1R-mediated neurite outgrowth. Overall, this study demonstrated that G alpha(o/i)-coupled CB1R triggers neurite outgrowth in Neuro-2A through the activation of a signaling network containing two pathways that bifurcate at Src and converge at Stat3. PMID- 16046414 TI - A novel platinum compound inhibits constitutive Stat3 signaling and induces cell cycle arrest and apoptosis of malignant cells. AB - Previous studies have established constitutive activation of Stat3 protein as one of the molecular changes required for tumorigenesis. To develop novel therapeutics for tumors harboring constitutively active Stat3, compounds from the NCI 2000 diversity set were evaluated for inhibition of Stat3 DNA-binding activity in vitro. Of these, a novel platinum (IV) compound, IS3 295, interacted with Stat3 and inhibited its binding to specific DNA-response elements. Further analysis suggested noncompetitive-type kinetics for the inhibition of Stat3 binding to DNA. In human and mouse tumor cell lines with constitutively active Stat3, IS3 295 selectively attenuated Stat3 signaling, thereby inducing cell growth arrest at G0/G1 phase and apoptosis. Moreover, in transformed cells, IS3 295 repressed expression of cyclin D1 and bcl-xL, two of the known Stat3 regulated genes that are overexpressed in malignant cells, suggesting that IS3 295 mediates anti-tumor cell activity in part by blocking Stat3-mediated sub version of cell growth and apoptotic signals. Together, our findings provide evidence for the inhibition of Stat3 activity and biological functions by IS3 295 through interaction with Stat3 protein. This study represents a significant advance in small molecule-based approaches to target Stat3 and suggests potential new applications for platinum (IV) complexes as modulators of the Stat3 pathway for cancer therapy. PMID- 16046416 TI - Portal imaging. PMID- 16046415 TI - A novel mechanism of G protein-dependent phosphorylation of vasodilator stimulated phosphoprotein. AB - Vasodilator-stimulated phosphoprotein (VASP) is a major substrate of protein kinase A (PKA). Here we described the novel mechanism of VASP phosphorylation via cAMP-independent PKA activation. We showed that in human umbilical vein endothelial cells (HUVECs) alpha-thrombin induced phosphorylation of VASP. Specific inhibition of Galpha13 protein by the RGS domain of a guanine nucleotide exchange factor, p115RhoGEF, inhibited thrombin-dependent phosphorylation of VASP. More importantly, Galpha13-induced VASP phosphorylation was dependent on activation of RhoA and mitogen-activated protein kinase kinase kinase, MEKK1, leading to the stimulation of the NF-kappaB signaling pathway. alpha-Thrombin dependent VASP phosphorylation was inhibited by small interfering RNA-mediated knockdown of RhoA, whereas Galpha13-dependent VASP phosphorylation was inhibited by a specific RhoA inhibitor botulinum toxin C3 and by a dominant negative mutant of MEKK1. We determined that Galpha13-dependent VASP phosphorylation was also inhibited by specific PKA inhibitors, PKI and H-89. In addition, the expression of phosphorylation-deficient IkappaB and pretreatment with the proteasome inhibitor MG-132 abolished Galpha13- and alpha-thrombin-induced VASP phosphorylation. In summary, we have described a novel pathway of Galpha13 induced VASP phosphorylation that involves activation of RhoA and MEKK1, phosphorylation and degradation of IkappaB, release of PKA catalytic subunit from the complex with IkappaB and NF-kappaB, and subsequent phosphorylation of VASP. PMID- 16046417 TI - Perception research in medical imaging. PMID- 16046418 TI - Review article: Acute serious and fatal reactions to contrast media: our current understanding. AB - Serious or fatal reactions to a contrast medium (CM) are unpredictable but fortunately rare. History of serious reaction to CM, bronchial asthma or multiple allergies increases the incidence of serious reactions by a factor of 5. Serious or fatal reaction to CM could be due to direct effect on basophils and mast cells or IgE mediated (type 1 hypersensitivity reaction). Activation of the kinin system leading to the formation of bradykinin could also be involved. Complement activation is unlikely to be a primary factor in initiating a serious reaction to CM. Avoiding CM administration in patients at high risk of serious reaction is advisable, but if the administration is deemed essential all precautions should be implemented and measures to treat serious reactions should be readily available. Oxygen supplementation, intravenous administration of physiological fluids and intramuscular injection of 0.5 ml adrenalin (1:1000) should be considered in the first line management of acute anaphylaxis. The ability to assess and treat serious CM reaction effectively is an essential skill that the radiologist should have and maintain. PMID- 16046419 TI - Radiological characterization of spinocerebellar ataxia type 6. AB - Spinocerebellar ataxia type 6 (SCA-6) is a rare, autosomal dominant neurodegenerative condition characterized by adult onset cerebellar ataxia and ocular movement disorders. The presentation is non-specific, and radiological characterization would be of diagnostic benefit. There is little published on the radiological appearances of SCA-6, and there are conflicting reports in the literature. We report the radiological findings in a group of 10 patients with SCA-6. PMID- 16046420 TI - Invasive pulmonary aspergillosis: frequency and meaning of the "hypodense sign" on unenhanced CT. AB - The purpose of this study was to establish the diagnostic value of central hypointensity ("hypodense sign") in lung consolidations or nodules, in severely immunocompromised or neutropenic patients, suspected of having invasive pulmonary aspergillosis (IPA), and to assess its recognition on unenhanced CT scans. Serial CT scans of the lung were retrospectively reviewed in 43 consecutive immunosuppressed patients with IPA, and assessed for the presence of the hypodense sign using standard mediastinal and lung windowing settings, as well as a special, narrower window setting (width 110-140 HU; level 15-40 HU). The temporal relationship between the occurrence of the first CT-finding suspicious of IPA and the appearance of the hypodense sign, as well as between this and the occurrence of the crescent sign, cavitation or reduction in lesion size, was evaluated. Additionally, CT-scans from 89 immunocompromised patients with viral (n=45) or bacterial (n=44) pneumonia, investigated in the same time period at our institution were reviewed, with respect to the presence of the "hypodense" sign. Unenhanced CT scans revealed the hypodense sign in 11 neutropenic patients and 2 severely immunocompromised patients, out of a total of 43 patients with IPA evaluated in this study (30.2%). The mean time between the appearance of the first CT-findings of IPA (large nodule or consolidation +/- positive halo sign) and the hypodense sign was 7.8 days, while the time interval between the hypodense sign and the occurrence of crescent sign, cavitation, or decrease of the lesion's size was 8.3 days. The hypodense sign did not occur in any of the patients with viral or bacterial pneumonia, in the control series. We consider the hypodense sign to be a supplementary tool in the diagnosis of IPA. Its sensitivity was low in our series, but the high specificity makes it valuable in predicting IPA, anticipating the occurrence of cavitation or crescent sign, which are considered specific, but late findings of IPA. The hypodense sign is recognizable also on unenhanced CT, when a narrower lung window setting is used. PMID- 16046421 TI - Differentiation of focal nodular hyperplasia and hepatocellular adenoma by contrast-enhanced ultrasound. AB - Non-invasive differentiation of focal nodular hyperplasia (FNH) and hepatocellular adenoma (HCA) is difficult. The aim of this study was to assess the accuracy of contrast-enhanced phase inversion ultrasound to differentiate between histologically proven FNH and HCA, analysing the arterial and (early) portal venous phase. 32 patients with histological proven FNH (n=24) or HCA (n=8) have been included in this prospective study. Examination technique: Siemens Elegra, phase inversion harmonic imaging (PIHI) with low mechanical index (MI)<0.2-0.3 using SonoVue (BR 1). The contrast enhancing tumour characteristics were evaluated during the hepatic arterial (starting 8-22 s) and early portal venous phase (starting 12-30 s). The image analysis was performed by three examiners. In 23 of 24 patients with FNH the contrast pattern revealed pronounced arterial and (early) portal venous enhancement. Homogeneous enhancement was detected during the hepatic arterial phase in all eight patients with HCA. In contrast to patients with FNH, no enhancement was seen during the portal venous phase. In conclusion, contrast-enhanced phase inversion ultrasound demonstrated pronounced arterial and portal venous enhancement in patients with focal nodular hyperplasia. In contrast, after homogeneous enhancement during hepatic arterial phase, no enhancement during hepatic portal venous phase was detected in patients with hepatocellular adenoma. Therefore, this technique might improve the functional characterization of benign hypervascular focal liver lesions. PMID- 16046422 TI - Imaging of diffuse metastatic and dystrophic pulmonary calcification in children after haematopoietic stem cell transplantation. AB - The authors describe three cases of diffuse pulmonary calcification; two metastatic in children with acute transitory renal failure and the other dystrophic in a child with leukaemia. All three patients underwent haematopoietic stem cell transplantation (HSCT). Chest radiographs disclosed diffuse calcification within the lungs. The distribution of this calcification was bilateral but asymmetric. Diagnosis was made in two cases by high resolution computed tomography (HRCT) and in one case by HRCT and bone scan. Radiological characteristics, scintigraphic features, pathological mechanism and clinical outcome of such pulmonary calcification are discussed. PMID- 16046423 TI - Diagnosis of ectopic gastric mucosa using 99Tcm-pertechnetate: spectrum of scintigraphic findings. AB - We highlight the spectrum of scintigraphic findings likely to be encountered in patients with ectopic gastric mucosa with illustrative cases for each such finding. 11 children (aged 4 months to 120 months, all males) underwent (99)Tc(m) pertechnetate scintigraphic evaluation for ectopic gastric mucosa. Functioning ectopic gastric mucosa was detected in Meckel's diverticulae in three patients, in small bowel duplications in four, in a gastric duplication in one, and in intrathoracic foregut duplication cysts in three. Ectopic functioning gastric mucosa in Meckel's diverticulum, and gastric duplication is visualized simultaneously with the stomach while in intestinal duplications tracer activity can be visualized in the dynamic sequence and even before gastric tracer visualization. In the three patients with intrathoracic duplication cysts, the functioning ectopic gastric mucosa was evident only in the delayed (99)Tc(m) pertechnetate images, much later than the visualization of stomach activity. Therefore, acquisition of delayed images are useful when the initial images are equivocal or negative in children with intrathoracic foregut duplication cysts. In addition, we suggest a hypothesis of a possible mechanism for the uptake of pertechnetate in ectopic gastric mucosa in some patients with intrathoracic forget duplication cysts. In conclusion, a variety of scintigraphic patterns may be found in patients with ectopic gastric mucosa undergoing (99)Tc(m) pertechnetate scintigraphy depending upon the location and size of the ectopic tissue. PMID- 16046424 TI - Pantak Therapax SXT 150: performance assessment and dose determination using IAEA TRS-398 protocol. AB - The performance assessment and beam characteristics of the Therapax SXT 150 unit, which encompass both low and medium-energy beams, were evaluated. Dose determination was carried out by implementing the International Atomic Energy Agency (IAEA) TRS-398 protocol and measuring all the dosimetric parameters in order to have a solid, consistent and reliable data set for the unit. Mechanical movements, interlocks and applicator characteristics agreed with specifications. The timer exhibited good accuracy and linearity. The output was very stable, with good repeatability, long-term reproducibility and no dependence on tube head orientation. The measured dosimetric parameters included beam first and second half-value layers (HVLs), absorbed dose rate to water under reference conditions, central axis depth dose distributions, output factors and beam profiles. Measured first HVLs agreed with comparable published data, but the homogeneity coefficients were low in comparison with typical values found in the literature. The timer error was significant for all filters and should be taken into consideration for the absorbed dose rate determination under reference conditions as well as for the calculation of treatment times. Percentage depth-dose (PDD) measurements are strongly recommended for each filter-applicator combination. The output factor definition of the IAEA TRS-398 protocol for medium-energy X-ray qualities involves the use of data that is difficult to measure. Beam profiles had small penumbras and good symmetry and flatness except for the lowest energy beam, for which a heel effect was observed. PMID- 16046425 TI - The effect of Butterworth and Metz reconstruction filters on volume and ejection fraction calculations with 99Tcm gated myocardial SPECT. AB - This study was carried out to measure the differences produced by change of reconstruction filter in calculations of left-ventricular end-diastolic volumes, end-systolic volumes, stroke-volumes and left-ventricular ejection-fractions from (99)Tc(m) Sestamibi (Bristol-Myers Squibb) gated myocardial perfusion SPECT studies. 30 patients had gated SPECT myocardial perfusion imaging at rest. The acquired projections were separately filtered with two filters, a low-pass filter (Butterworth) and an edge-enhancement filter (Metz). Each study was then further processed to determine left-ventricular end-diastolic volume, end-systolic volume, stroke volume and ejection fraction, and to assess defect size. The results for each patient with the two filters were compared. Calculated end diastolic volumes, end-systolic volumes and left-ventricular ejection fractions, for each filter, were well correlated. Stroke volumes showed worse correlation. The differences between left-ventricular ejection-fractions, end-diastolic volumes and end-systolic volumes were statistically significant. There was no significant difference in stroke volumes. Ejection fractions were inversely correlated with defect size, but change in ejection fraction due to filter was not. End-diastolic and end-systolic volumes were correlated with defect size, but change in volumes due to filter was not. Thus the results for changes produced by choice of filter are not dependent on defect size. Using different reconstruction pre-filters in gated myocardial perfusion SPECT significantly changes the results of calculations of physiological parameters. Each centre should be consistent in the use of filters as this may affect the clinical consequences of the result. PMID- 16046426 TI - An assessment of clinically optimal gold marker length and diameter for pelvic radiotherapy verification using an amorphous silicon flat panel electronic portal imaging device. AB - Verification of target organ position is essential for the accurate delivery of conformal radiotherapy. Megavoltage electronic portal imaging with flat panel amorphous silicon detectors delivers high quality images that can be used for verification of bony landmark position. Gold markers implanted into the target organ can be visualized and used as a surrogate of actual organ position. On-line compensation for marker displacement, by adjusting patient position, can reduce geometric errors associated with radiation delivery. This study assesses the optimal marker length and diameter to be used with an amorphous silicon (a-Si) flat panel detector and electronic portal images (EPIs), prior to implementation of a clinical programme of gold marker insertion in prostate cancer patients. Seven marker sizes varying from 3 mm to 8 mm in length and 0.8 mm to 1.1 mm in diameter were investigated in a group of patients undergoing pelvic radiotherapy using an 8 MV Elekta SL20 linear accelerator. Markers were placed on the skin entry and exit sites of the treatment beam and EPIs in both lateral and anterior pelvic views were acquired. Three observers independently assessed visibility success and failure using a subjective scoring system. Markers less than 5 mm in length or 0.9 mm in diameter were poorly visualized (<70% visualization success in lateral EPIs). The marker measuring 0.9 mm x 5 mm appears to be clinically optimal in pelvic radiotherapy patients (80% visualization success in lateral EPIs) and will be used for actual organ implantation. PMID- 16046427 TI - Short communication: Setup variations in locoregional radiotherapy for breast cancer: an electronic portal imaging study. AB - Recent trials demonstrating a survival benefit with locoregional radiotherapy (LRRT) to the chest wall and regional nodes in women with node-positive breast cancer have led to increased use of complex techniques to match three or more radiation fields, but information on setup reproducibility with LRRT for breast cancer is scarce. This study reports the magnitude and directions of random and systematic deviations in LRRT for breast cancer using an offline electronic portal imaging verification protocol. Electronic portal images (EPIs) of 46 consecutive women treated with LRRT for breast cancer from March 2001 to February 2002 with LRRT were analysed. Comparisons of EPIs to the corresponding digitally reconstructed radiographs were performed offline with anatomy matching. Displacements in mm were recorded in the superior-inferior (SI), medial-lateral (ML), and anterior-posterior (AP) directions. Random errors ranged from 2.0 mm to 2.5 mm for the breast/chest wall tangential treatments and 2.3 mm to 3.9 mm for the supraclavicular nodal treatments. Systematic errors occurred to a greater degree in the AP direction for the tangential fields and in the ML direction for the supraclavicular field. Displacements of > or =10 mm were found in 1.2% of breast/chest wall tangential treatments and in 6.2% of supraclavicular nodal treatments. These data demonstrate that EPI is a useful tool to verify setup reproducibility in LRRT for breast cancer. PMID- 16046428 TI - Short communication: A printed image quality test phantom for mammography. AB - This communication describes a novel design for a mammographic image quality test phantom, the final design of which was produced as a radiographer weekly quality assurance phantom for breast screening and symptomatic mammography. The phantom is based on low contrast test features which are built up by superimposing sheets of Mylar overhead projector transparency, on which the test features are printed using a standard LaserJet printer. The required radiation contrast at mammographic energies is produced by the approximately 50% by weight component of iron oxide (Fe(3)O(4)) present in the toner. An easily replicated design of mammographic image quality phantom based on LaserJet printed test features is described. Approximately 40 of these phantoms were constructed, and these have been used successfully for 5 years in both breast screening and symptomatic mammography. The phantom design offers a performance similar to much more expensive mammographic contrast-detail phantoms, but suffers from the disadvantage that high contrast resolution bar patterns cannot be produced using the standard printing process. PMID- 16046429 TI - Short communication: A method for verified access when using soft copy display. AB - Soft copy display is a rapidly developing area. To date, most soft copy systems can be classed by their application, e.g. review or reporting. With technology convergence this distinction is becoming less defined by the hardware and more defined by the software functionality. Although it is accepted that routine quality assurance should be conducted on soft copy monitors, this would be logistically difficult to achieve if any monitor within a hospital could be used for image review or reporting. This work proposes a simple psychophysical check to ensure optimal display performance before viewing software can be run. This is in the form of a challenge/response code constructed from letters just above the threshold of detection. This verified login would act as a portal to launching the image viewing software. The developed system was tested on three different types of monitor and five observers. Results indicate that the verified login was able to control access for displays below the optimal settings but was not as sensitive for adjustments above the optimum. However it is believed this is still of value as the lower presentation will compress the display gamma curve and reduce detail contrast. It also provides a minimum level of audit and quality control that might otherwise be missing. PMID- 16046430 TI - Case report: Foreign body granuloma mimicking liver metastasis. AB - In this study, foreign body granuloma mimicking liver metastasis diagnosed on routine follow-up examination in a 41-year-old woman with rectal adenocarcinoma is reported. To our knowledge, this is the first study in English-language literature reporting foreign body granuloma indistinguishable from liver metastasis on radiological examination. PMID- 16046431 TI - Case report: MR imaging features of endometriosis at the umbilicus. AB - The radiological features of extrapelvic endometriosis at the umbilicus with large ovarian endometriomas are described. In this patient, the umbilical lesion appears as hypointense on T(1) weighted and T(2) weighted MR images. The MR characteristics of endometriosis at the umbilicus are compared with those found within the pelvis. PMID- 16046432 TI - Case report: Gluteal injection site granulomas: false positive finding on FDG-PET in patients with non-small cell lung cancer. AB - Positron-emission-tomography (PET) with fludeoxyglucose F-18 ([(18)F] fluoro-2 deoxy-D-glucose, FDG) has become an established imaging modality in patients with lung cancer for mediastinal lymph node staging and the detection of extrathoracic metastases. However, tracer accumulations are not limited to malignant tissue but are also found in muscles and benign inflammatory processes. We report on two patients with lung cancer in whom FDG-PET revealed suspicious tracer accumulations in the buttock. Ultrasound (US) revealed a hyperechogenic nodule with poorly defined margins in both patients. On specific inquiry both patients reported on repeated "intramuscular" gluteal injections. Histology after US guided biopsy showed an accumulation of macrophages within fibrous tissue, compatible with injection site granulomas. The reported cases underline that (18)F-FDG may accumulate in benign, ancillary processes that have to be distinguished from distant metastases. Tracer accumulation in the buttocks may be highly suggestive of injection site granulomas, especially if the patient reports on "intramuscular" injections. In this setting, US is a widely available modality to distinguish metastasis from adipose tissue necrosis. PMID- 16046433 TI - Case report: Fatal acute exacerbation of usual interstitial pneumonia in ulcerative colitis. AB - Pulmonary involvement in ulcerative colitis may manifest as a variety of disorders. Ulcerative colitis-related interstitial lung disease is exceedingly rare and has been reported to be steroid-responsive. We describe the first case of a patient with acute exacerbation of ulcerative colitis-induced usual interstitial pneumonia, who did not respond to corticosteroid therapy and died 12 weeks after the onset of pulmonary symptoms. Early recognition of pulmonary disease in patients with ulcerative colitis is necessary to initiate further diagnostic work-up and may aid treatment decisions. PMID- 16046434 TI - Case report: Portal and systemic venous gas in a patient after lumbar puncture. AB - The presence of portal and systemic venous gas is traditionally regarded as an ominous radiological sign indicating a grave prognosis. With advances in imaging technology, the incidence of its detection has increased along with its association with clinically benign disorders. We present a young patient with systemic and portal venous gas after traumatic lumbar puncture. PMID- 16046435 TI - Abandoning linear no threshold. PMID- 16046436 TI - Ultrasound-guided catheter removal. PMID- 16046437 TI - Case of the Month: A patient with acute abdominal pain and a peritoneal mass on ultrasound and multidetector CT. PMID- 16046439 TI - Significant variation in mortality and functional outcome after acute ischaemic stroke between Western countries: data from the tinzaparin in acute ischaemic stroke trial (TAIST). AB - BACKGROUND: The medical care of patients with acute stroke varies considerably between countries. This could lead to measurable differences in mortality and functional outcome. OBJECTIVE: To compare case mix, clinical management, and functional outcome in stroke between 11 countries. METHODS: All 1484 patients from 11 countries who were enrolled into the tinzaparin in acute ischaemic stroke trial (TAIST) were included in this substudy. Information collected prospectively on demographics, risk factors, clinical features, measures of service quality (for example, admission to a stroke unit), and outcome were assessed. Outcomes were adjusted for treatment assignment, case mix, and service relative to the British Isles. RESULTS: Differences in case mix (mostly minor) and clinical service (many of prognostic relevance) were present between the countries. Significant differences in outcome were present between the countries. When assessed by geographical region, death or dependency were lower in North America (odds ratio (OR) adjusted for treatment group only = 0.52 (95% confidence interval, 0.39 to 0.71) and north west Europe (OR = 0.54 (0.37 to 0.78)) relative to the British Isles; similar reductions were found when adjustments were made for 11 case mix variables and five service quality measures. Similarly, case fatality rates were lower in North America (OR = 0.44 (0.30 to 0.66)) and Scandinavia (OR = 0.50 (0.33 to 0.74)) relative to the British Isles, whether crude or adjusted for case mix and service quality. CONCLUSIONS: Both functional outcome and case fatality vary considerably between countries, even when adjusted for prognostic case mix variables and measures of good stroke care. Differing health care systems and the management of patients with acute stroke may contribute to these findings. PMID- 16046440 TI - Household based treatment of drinking water with flocculant-disinfectant for preventing diarrhoea in areas with turbid source water in rural western Kenya: cluster randomised controlled trial. AB - OBJECTIVE: To compare the effect on prevalence of diarrhoea and mortality of household based treatment of drinking water with flocculant-disinfectant, sodium hypochlorite, and standard practices in areas with turbid water source in Africa. DESIGN: Cluster randomised controlled trial over 20 weeks. SETTING: Family compounds, each containing several houses, in rural western Kenya. PARTICIPANTS: 6650 people in 605 family compounds. INTERVENTION: Water treatment: flocculant disinfectant, sodium hypochlorite, and usual practice (control). MAIN OUTCOME MEASURES: Prevalence of diarrhoea and all cause mortality. Escherichia coli concentration, free residual chlorine concentration, and turbidity in household drinking water as surrogates for effectiveness of water treatment. RESULTS: In children < 2 years old, compared with those in the control compounds, the absolute difference in prevalence of diarrhoea was -25% in the flocculant disinfectant arm (95% confidence interval -40 to -5) and -17% in the sodium hypochlorite arm (-34 to 4). In all age groups compared with control, the absolute difference in prevalence was -19% in the flocculant-disinfectant arm ( 34 to -2) and -26% in the sodium hypochlorite arm (-39 to -9). There were significantly fewer deaths in the intervention compounds than in the control compounds (relative risk of death 0.58, P = 0.036). Fourteen per cent of water samples from control compounds had E coli concentrations < 1 CFU/100 ml compared with 82% in flocculant-disinfectant and 78% in sodium hypochlorite compounds. The mean turbidity of drinking water was 8 nephelometric turbidity units (NTU) in flocculant-disinfectant households, compared with 55 NTU in the two other compounds (P < 0.001). CONCLUSIONS: In areas of turbid water, flocculant disinfectant was associated with a significant reduction in diarrhoea among children < 2 years. This health benefit, combined with a significant reduction in turbidity, suggests that the flocculant-disinfectant is well suited to areas with highly contaminated and turbid water. PMID- 16046441 TI - Scale-freeness and biological networks. AB - The notion of scale-freeness and its prevalence in both natural and artificial networks have recently attracted much attention. The concept of scale-freeness is enthusiastically applied to almost any conceivable network, usually with affirmative conclusions. Well-known scale-free examples include the internet, electric lines among power plants, the co-starring of movie actors, the co authorship of researchers, food webs, and neural, protein-protein interactional, genetic, and metabolic networks. The purpose of this review is to clarify the relationship between scale-freeness and power-law distribution, and to assess critically the previous related works, especially on biological networks. In addition, I will focus on the close relationship between power-law distribution and lognormal distribution to show that power-law distribution is not a special characteristic of natural selection. PMID- 16046443 TI - From SRY to SOX9: mammalian testis differentiation. AB - Sry (sex-determining region on the Y chromosome) is a master gene that initiates testis differentiation of the bipotential indifferent gonad in mammals. In mice, Sry expression is transiently activated in a center-to-pole wave along the anteroposterior (AP) axis of developing XY gonads. Shortly after the onset of Sry activation, Sox9 (Sry-related HMG box-9), a fundamental testis-differentiation gene common to all vertebrates, is also activated in a center-to-pole pattern similar to the initial Sry expression profile. Several male-specific cellular events, such as glycogenesis, coelomic epithelium proliferation, mesonephric migration and vasculogenesis, are induced in XY gonads following the onset of Sry and Sox9 expression. This paper mainly focuses on recent advances in elucidating the regulatory mechanisms of Sry and Sox9 expression and male-specific cellular events immediately downstream of SRY action during the initial phases of testis differentiation. PMID- 16046442 TI - Temperature, genes, and sex: a comparative view of sex determination in Trachemys scripta and Mus musculus. AB - Sex determination, the step at which differentiation of males and females is initiated in the embryo, is of central importance to the propagation of species. There is a remarkable diversity of mechanisms by which sex determination is accomplished. In general these mechanisms fall into two categories: Genetic Sex Determination (GSD), which depends on genetic differences between the sexes, and Environmental Sex Determination (ESD), which depends on extrinsic cues. In this review we will consider these two means of determining sex with particular emphasis on two species: a species that depends on GSD, Mus musculus, and a species that depends on ESD, Trachemys scripta. Because the structural organization of the adult testis and ovary is very similar across vertebrates, most biologists had expected that the pathways downstream of the sex-determining switch would be conserved. However, emerging data indicate that not only are the initial sex determining mechanisms different, but the downstream pathways and morphogenetic events leading to the development of a testis or ovary also are different. PMID- 16046444 TI - Probing of specific binding of synthetic sulfonylurea with the insulinoma cell line MIN6. AB - To overcome the limitation of conventional sulfonylurea (SU) for investigation of biological mechanisms related to KATP channels, a hypoglycemic sulfonylurea (SU) was conjugated with a non-reducing glucose bearing polystyrene (PS) derivative to provide enhanced interaction with an insulinoma cell line (MIN6). The specific interaction between the SU (K+ channel closer)-conjugated copolymer and MIN6 cells was confirmed by confocal laser microscopic images using rhodamine B isothiocyanate (RITC)-labeled SU-conjugated polymer, which revealed the specific interaction between SU-conjugated polymer and MIN6 cells. Moreover, the location of labeled polymer and the site of Ca2+ ion mobilization obtained from the same MIN6 cells were identical. Based on the specificity and insulinotropic activity, the SU-conjugated polymer is expected to be useful tool for the study of biological mechanisms of KATP channels. PMID- 16046445 TI - Role of the N-terminal domain of endoinulinase from Arthrobacter sp. S37 in regulation of enzyme catalysis. AB - Endoinulinase from Arthrobacter sp. S37 (EnIA), a member of the glycoside hydrolase family 32, is unique in that, unlike other members of the family, it contains a 250-residue N-terminal domain including a "laminin-G like jelly-roll" fold. This unique N-terminal domain is here suggested to be involved in dimerization and catalysis. The essentially inactive nature of enzymes produced by N-terminal truncation (Delta15, Delta45, Delta70, and Delta250) supported the pivotal role of this unique domain in catalysis and the need for its structural integrity. Significant reductions in the enzyme efficiency (kcat/Km) were observed when mutations were introduced at highly conserved tryptophan residues (Trp75 and Trp141) in the laminin-G like jelly-roll fold, implying their involvement in catalysis. Results from size-exclusion chromatography of the native and chimeric enzymes in the presence and absence of the domain suggested that the N-terminal domain could mediate dimerization. PMID- 16046446 TI - Roles of three well-conserved arginine residues in mediating the catalytic activity of tobacco acetohydroxy acid synthase. AB - Acetohydroxy acid synthase (AHAS, EC 2.2.1.6; also known as acetolactate synthase, ALS) catalyzes the first common step in the biosynthesis of valine, leucine, and isoleucine in plants and microorganisms. AHAS is the target of several classes of herbicides. In the present study, the role of three well conserved arginine residues (R141, R372, and R376) in tobacco AHAS was determined by site-directed mutagenesis. The mutated enzymes, referred to as R141A, R141F, and R376F, were inactive and unable to bind to the cofactor, FAD. The inactive mutants had the same secondary structure as that of the wild type. The mutants R141K, R372F, and R376K exhibited much lower specific activities than the wild type, and moderate resistance to herbicides such as Londax, Cadre, and/or TP. The mutation R141K showed a strong reduction in activation efficiency by ThDP, while the mutations R372K and R376K showed a strong reductions in activation efficiency by FAD in comparison to the wild type enzyme. Taking into account the data presented here and the homology model constructed previously [Le et al. (2004) Biochem. Biophys. Res. Commun. 317, 930-938], it is suggested that the three amino acid residues studied (R141, R372, and R376) are located essentially at the enzyme active site, and, furthermore, that residues R372 and R376 are possibly responsible for the binding of the enzyme to FAD. PMID- 16046447 TI - Ribosomal proteins cross-linked to the initiator AUG codon of a mRNA in the translation initiation complex by UV-irradiation. AB - Eukaryotic ribosomal proteins constituting the binding site for the initiator codon AUG on the ribosome at the translation initiation step were investigated by UV-induced cross-linking between protein and mRNA. The 80S-initiation complex was formed in a rabbit reticulocyte cell-free system in the presence of sparsomycin with radiolabeled Omega-fragment as a template, which was a 73-base 5'-leader sequence of tobacco mosaic virus RNA having AUG at the extreme 3'-terminal end and extended with 32pCp. Two radioactive peaks were sedimented by sucrose gradient centrifugation, one being the 80S initiation complex formed at the 3' terminal AUG codon, and the other presumably a "disome" with an additional 80S ribosome bound at an upstream AUU codon, formed when Omega-fragment was incubated with sparsomycin [Filipowicz and Henni (1979) Proc. Natl. Acad. Sci. USA 76, 3111 3115]. Cross-links between ribosomal proteins and the radiolabeled Omega-fragment were induced in situ by UV-irradiation at 254 nm. After extensive nuclease digestion of the complexes, ribosomal proteins were separated by two-dimensional gel electrophoresis. Autoradiography identified the proteins S7, S10, S25, S29, and L5 of the 80S initiation complex and S7, S25, S29 and L5 of that in the disome as 32P-labeled proteins. Together with the results of cross-linking experiments of other investigators and recently solved crystal structures of prokaryotic ribosomes, the spatial arrangement of eukaryotic ribosomal proteins at the AUG-binding domain is discussed. PMID- 16046448 TI - Sphingosine 1-phosphate-related metabolism in the blood vessel. AB - Sphingosine 1-phosphate (Sph-1-P) is a bioactive lipid released from activated platelets and plays an important role in vascular biology. In this study, we investigated Sph-1-P-related metabolism in the blood vessel, mainly using radio labeled Sph and Sph-1-P. Sph was metabolically stable in the plasma, while it was converted into Sph-1-P in the presence of activated platelets. When the mixture of Sph-1-P and plasma was fractionated on a gel-filtration column, all Sph-1-P co eluted with protein fractions that coincide with lipoproteins and albumin by agarose gel electrophoresis. When evaluated by polyacrylamide gel electrophoresis, 7.2 +/- 3.8%, 53.3 +/- 6.4%, and 39.5 +/- 7.9% of the radioactivity of Sph-1-P added to plasma was recovered in the low-density lipoprotein (LDL), high-density lipoprotein (HDL), and albumin fractions, respectively. On the other hand, 5.2 +/- 3.2%, 38.4 +/- 5.5%, and 56.3 +/- 5.7% of the radioactivity of Sph-1-P converted from Sph in collagen-stimulated platelets and released into the plasma was recovered in the LDL, HDL, and albumin fractions, respectively. When Sph-1-P release from activated platelets was examined, a stronger response was observed in the presence of albumin than lipoproteins, suggesting efficient Sph-1-P extraction from platelets by albumin. Finally, Sph-1-P, which is stable in the plasma, was markedly degraded by an ectophosphatase activity in the presence of vascular endothelial cells or in whole blood. Although Sph-1-P is stable in the plasma, it is likely that the level of this bioactive lipid is dynamically controlled by various factors including release from platelets, distribution among plasma proteins, and degradation by ectophosphatase. PMID- 16046449 TI - Nitrated and oxidized products of a single tryptophan residue in human Cu,Zn superoxide dismutase treated with either peroxynitrite-carbon dioxide or myeloperoxidase-hydrogen peroxide-nitrite. AB - We reported previously that a single tryptophan residue, Trp32, in human Cu,Zn superoxide dismutase is specifically modified by peroxynitrite-CO2 [Yamakura et al. (2001) Biochim. Biophys. Acta 1548, 38-46]. In this study, we modified Cu,Zn superoxide dismutase by using a combination of myeloperoxidase, hydrogen peroxide, and nitrite. The modified enzyme showed no loss of copper and zinc, and 15% less enzymatic activity. Trp32 was the only significant amino acid lost. After trypsin digestion of the modified SOD with peroxynitrite-CO2 and the myeloperoxidase system, six newly appearing peptides containing tryptophan derivatives were observed on microLC-ESI-Q-TOF mass analyses and HPLC with a photodiode-array detector. The derivatives of the tryptophan residue exhibiting mass increases of 4, 16 (2 peaks), 32, 45 (major), and 45 Da (minor) were identified as kynurenine, oxindole-3-alanine and its derivatives, dihydroxytryptophan, 6-nitrotryptophan and 5-nitrotryptophan, respectively. We further identified 6-nitrotryptophan from the 1H-NMR spectrum for the pronase digested product and calculated the yield of 6-nitrotryptophan as being about 30% for each of the modification methods. The tryptophan residue in the modified human Cu,Zn-superoxide dismutase gave the same spectra for the products including 6-nitrotryptophan as the major nitrated product with the two different modification systems. PMID- 16046450 TI - Purification and characterization of a hemoglobin degrading aspartic protease from the malarial parasite Plasmodium vivax. AB - Aspartic proteases of human malarial parasites are thought to play key roles in essential pathways of merozoite release, invasion and host cell hemoglobin degradation during the intraerythrocytic stages of their life cycle. Therefore, we have purified and characterized Plasmodium vivax aspartic protease, to determine if this enzyme can be used as potential drug target/immunogen, and its inhibitors as potential antimalarial drug. The P. vivax aspartic protease has been purified by a combination of ion exchange and size exclusion chromatographies and HPLC. Its properties were examined in order to define a role in the hemoglobin degradation process. The purified enzyme migrated as a single band on native PAGE and SDS/PAGE with a molecular mass of 40 kDa. Gelatin zymogram analyses revealed a clear zone of proteolytic activity corresponding to the band obtained on native PAGE and SDS/PAGE. The enzyme has an optimal pH of 4.0 and exhibits its highest activity at 37 degrees C. The enzyme is inhibited by pepstatin, but not by other inhibitors including o-phenanthroline, EDTA, PMSF or E-64, supporting its designation as an aspartic protease; its IC50 value was found to be 3.0 microM. A Lineweaver Burk double reciprocal plot with pepstatin shows that the inhibition is competitive with respect to the substrate. Ca2+ and Mg2+ ions enhance the protease activity, whereas Cu2+ and Hg2+ ions were found to be inhibitory. The pivotal role of aspartic protease in initiating hemoglobin degradation in P. vivax malaria parasite is also demonstrated. PMID- 16046451 TI - Improved estimation of the secondary structures of proteins by vacuum-ultraviolet circular dichroism spectroscopy. AB - The vacuum-ultraviolet circular dichroism (VUVCD) spectra of 16 globular proteins (insulin, lactate dehydrogenase, glucose isomerase, lipase, conalbumin, transferrin, catalase, subtilisin A, alpha-amylase, staphylococcal nuclease, papain, thioredoxin, carbonic anhydrase, elastase, avidin, and xylanase) were successfully measured in aqueous solutions at 25 degrees C from 260 to 160 nm under a high vacuum using a synchrotron-radiation VUVCD spectrophotometer. These proteins exhibited characteristic CD spectra below 190 nm that were related to their different secondary structures, which could not be detected with a conventional CD spectrophotometer. The component spectra of alpha-helices, beta strands, turns, and unordered structures were obtained by deconvolution analysis of the VUVCD spectra of 31 reference proteins including the 15 proteins reported in our previous paper [Matsuo, K. et al. (2004) J. Biochem. 135, 405-411]. Prediction of the secondary-structure contents using the SELCON3 program was greatly improved, especially for alpha-helices, by extending the short-wavelength limit of CD spectra to 160 nm and by increasing the number of reference proteins. The numbers of alpha-helix and beta-strand segments, which were calculated from the distorted alpha-helix and beta-strand contents, were close to those obtained on X-ray crystallography. These results demonstrate the usefulness of synchrotron radiation VUVCD spectroscopy for the secondary structure analysis of proteins. PMID- 16046452 TI - Expression in Escherichia coli, purification and characterization of Thermoanaerobacter tengcongensis ribosome recycling factor. AB - A very promising approach to understanding the mechanism of protein thermostability is to investigate the structure-function relationship of homologous proteins with different thermostabilities. Ribosome recycling factor (RRF), which is an essential factor for protein synthesis in bacteria, may be a good candidate for such study. In this report, a ribosome recycling factor from Thermoanaerobacter tengcongensis was expressed and characterized. This protein contains 184 residues, shows 51.4% identity to that of Escherichia coli RRF, and has very strong antigenic cross-reactivity with antibody to E. coli RRF. In vivo activity assay shows that weak residual activity may remain in TteRRF in E. coli cells. Circular dichroism spectral analysis shows that TteRRF has a very similar secondary structure to that of E. coli RRF, implying that they have similar tertiary structures. However, their thermostabilities are significantly different. To find which domain of RRF is mainly responsible for maintaining stability, TteDI/EcoDII and EcoDI/TteDII RRF chimeras were created. Their domain I and domain II are from E. coli and T. tengcongensis RRFs, respectively. The results of GdnHCl and heat induced denaturation of the chimeric RRFs suggest that the domain I plays a major role in maintaining the stability of the RRF molecule. PMID- 16046453 TI - Removal of tightly bound ADP induces distinct structural changes of the two tryptophan-containing regions of the ncd motor domain. AB - ncd is a molecular motor belonging to the kinesin superfamily. In solution, it is a homo-dimer of a 700 amino acid polypeptide. The C-terminus of each polypeptide forms a globular domain of about 40 kDa, the motor domain with ATPase activity. The ATPase site of the motor domain of kinesin family members, including ncd, binds ADP tightly, the release of which is facilitated by microtubules during the mechanochemical ATPase cycle. Previously, we studied the spectroscopic characteristics of the ncd motor domain, focusing on interactions of the transition-moment-dipoles between ADP and aromatic amino acid side chains using circular dichroism (CD) spectroscopy. In the present study, we generated several ncd motor domain mutants. In each, a tryptophanyl or specific tyrosyl residue was mutated. We found that Trp370 and Tyr442, the latter of which stacks directly with the adenine moiety of bound ADP, caused the bound ADP to exhibit peculiar CD signals. In addition, fluorescence measurements revealed that Trp370, but not Trp473, was responsible for the emission intensity change depending on the presence or absence of bound ADP. This fluorescence result implies that the structural change induced at the ADP-binding site (on the release of the ADP) is transmitted to the region that includes Trp370, which is relatively close to the ADP-binding site but not in direct contact with the ADP-binding region. In contrast, Trp473 in the region that is in contact with the alpha-helical coiled coil stalk did not experience the structural changes caused on removal of ADP. The distinct behavior of these two tryptophanyl residues suggests that the ncd motor domain has a bifacial architecture made up of a relatively deformable side including the nucleotide binding site and a more rigid one. PMID- 16046454 TI - Hindlimb casting decreases muscle mass in part by proteasome-dependent proteolysis but independent of protein synthesis. AB - The hypothesis of the present study was that rats subjected to short-term unilateral hindlimb immobilization would incur skeletal muscle wasting and concomitant alterations in protein synthesis, controllers of translation, and indexes of protein degradation. Rats were unilaterally casted for 1, 3, or 5 days to avoid complications associated with other disuse models. In the casted limb, gastrocnemius wet weight decreased 12% after 3 days and thereafter remained constant. In contrast, the contralateral control leg displayed a steady growth rate over time. The rate of protein synthesis and translational efficiency were unchanged in the immobilized muscle at day 5. The total amount and phosphorylation state of regulators of translational initiation and elongation were unaltered. The mRNA contents of polyubiquitin and the ubiquitin ligases muscle atrophy F-box (MAFbx)/Atrogin-1 and muscle RING finger 1 (MuRF1) were elevated in immobilized muscle at all time points, with peak expression occurring at day 3. Daily injection of the type II glucocorticoid receptor antagonist RU 486 did not prevent decreases in gastrocnemius wet weight nor increases in mRNA for MAFbx/Atrogin-1 and MuRF1. However, in vivo administration of the proteasome inhibitor Velcade prevented 53% of wet weight loss associated with 3 days of immobilization. These data suggest that the loss of skeletal muscle mass in this model of disuse appears to be glucocorticoid independent, can be partially rescued with a potent proteasome inhibitor, and is associated with enhanced mRNA expression of multiple factors that contribute to ubiquitin- proteasome-dependent degradation and are likely to control the remodeling of immobilized skeletal muscle during atrophy. PMID- 16046455 TI - Effects of 2 wk of GH administration on 24-h indirect calorimetry in young, healthy, lean men. AB - The present study was designed as a randomized, double-blind placebo (Plc) controlled study to determine the effect of 2 wk of growth hormone administration (GH-adm.) on energy expenditure (EE) and substrate oxidation in healthy humans. Sixteen young healthy men were divided into two groups. The study consisted of two 24-h measurements (indirect calorimetry), separated by 2 wk of either Plc or GH injections (6 IU/day). At baseline, no significant differences were observed between the two groups in any of the measured anthropometric, hormonal, or metabolic parameters, neither did the parameters change over time in the Plc group. GH-adm. resulted in a 4.4% increase in 24-h EE (P < 0.05) and an increase in fat oxidation by 29% (P < 0.05). However, a decrease in the respiratory quotient was only observed in the postabsorptive phase after an overnight fast (0.84 +/- 0.1 to 0.79 +/- 0.1, P < 0.05). Furthermore, lean body mass (LBM) was increased by GH-adm. only [62.8 +/- 2.5 kg (baseline) vs. 64.7 +/- 2.4 kg (after), P < 0.001]. In conclusion, GH-adm. increases 24-h EE, which may be partly explained by increased LBM. Furthermore, GH-adm. stimulates fat combustion, especially in the postabsorptive state. PMID- 16046456 TI - Leptin inhibits hypothalamic Npy and Agrp gene expression via a mechanism that requires phosphatidylinositol 3-OH-kinase signaling. AB - Phosphatidylinositol 3-OH-kinase (PI3K) and STAT3 are signal transduction molecules activated by leptin in brain areas controlling food intake. To investigate their role in leptin-mediated inhibition of hypothalamic neuropeptide Y (Npy) and agouti-related peptide (Agrp) gene expression, male Sprague-Dawley rats (n = 5/group) were either fed ad libitum or subjected to a 52-h fast. At 12 h intervals, the PI3K inhibitor LY-294002 (LY, 1 nmol) or vehicle was injected intracerebroventricularly (ICV) as a pretreatment, followed 1 h later by leptin (3 microg icv) or vehicle. Fasting increased hypothalamic Npy and Agrp mRNA levels (P < 0.05), and ICV leptin administration prevented this increase. As predicted, LY pretreatment blocked this inhibitory effect of leptin, such that Npy and Agrp levels in LY-leptin-treated animals were similar to fasted controls. By comparison, leptin-mediated activation of hypothalamic STAT3 signaling, as measured by induction of both phospho-STAT3 immunohistochemistry and suppressor of cytokine signaling-3 (Socs3) mRNA, was not significantly attenuated by ICV LY pretreatment. Because NPY/AgRP neurons project to the hypothalamic paraventricular nucleus (PVN), we next investigated whether leptin activation of PVN neurons is similarly PI3K dependent. Compared with vehicle, leptin increased the number of c-Fos positive cells within the parvocellular PVN (P = 0.001), and LY pretreatment attenuated this effect by 35% (P = 0.043). We conclude that leptin requires intact PI3K signaling both to inhibit hypothalamic Npy and Agrp gene expression and activate neurons within the PVN. In addition, these data suggest that leptin activation of STAT3 is insufficient to inhibit expression of Npy or Agrp in the absence of PI3K signaling. PMID- 16046457 TI - 5-Aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside renders glucose output by the liver of the dog insensitive to a pharmacological increment in insulin. AB - This study aimed to test whether stimulation of net hepatic glucose output (NHGO) by increased concentrations of the AMP analog, 5-aminoimidazole-4-carboxamide-1 beta-d-ribosyl-5-monophosphate, can be suppressed by pharmacological insulin levels. Dogs had sampling (artery, portal vein, hepatic vein) and infusion (vena cava, portal vein) catheters and flow probes (hepatic artery, portal vein) implanted >16 days before study. Protocols consisted of equilibration (-130 to 30 min), basal (-30 to 0 min), and hyperinsulinemic-euglycemic (0-150 min) periods. At time (t) = 0 min, somatostatin was infused, and basal glucagon was replaced via the portal vein. Insulin was infused in the portal vein at either 2 (INS2) or 5 (INS5) mU.kg(-1).min(-1). At t = 60 min, 1 mg.kg(-1).min(-1) portal venous 5-aminoimidazole-4-carboxamide-1-beta-d-ribofuranoside (AICAR) infusion was initiated. Arterial insulin rose approximately 9- and approximately 27-fold in INS2 and INS5, respectively. Glucagon, catecholamines, and cortisol did not change throughout the study. NHGO was completely suppressed before t = 60 min. Intraportal AICAR stimulated NHGO by 1.9 +/- 0.5 and 2.0 +/- 0.5 mg.kg(-1).min( 1) in INS2 and INS5, respectively. AICAR stimulated tracer-determined endogenous glucose production similarly in both groups. Intraportal AICAR infusion significantly increased hepatic acetyl-CoA carboxylase (ACC, Ser(79)) phosphorylation in INS2. Hepatic ACC (Ser(79)) phosphorylation, however, was not increased in INS5. Thus intraportal AICAR infusion renders hepatic glucose output insensitive to pharmacological insulin. The effectiveness of AICAR in countering the suppressive effect of pharmacological insulin on NHGO occurs even though AICAR-stimulated ACC phosphorylation is completely blocked. PMID- 16046458 TI - Somatostatin and dopamine receptor expression in lung carcinoma cells and effects of chimeric somatostatin-dopamine molecules on cell proliferation. AB - To study somatostatin/dopamine (SS/D) synergy in a human cell system constitutively expressing SS and D receptors (SSR and DR, respectively), we characterized the expression of SSR and DR subtypes in the non-small-cell lung cancer line Calu-6, and then we evaluated the effect on cell proliferation of SS/D chimeric molecules (BIM-23A387 and BIM-23A370), which bind with high affinity both sst(2) and D(2)R, and compared the results with those obtained by using SS-14 and subtype-selective SS analogs (SSA) and D agonists (DA). Because Calu-6 cells produce insulin-like growth factor (IGF) and IGF-binding protein (IGFBP) peptides, which play a role in the autocrine/paracrine control of cell growth, we also investigated the effects of chimeric compounds on secretion and expression of IGF system components. Relative high levels of sst(2) and the long isoform of the D(2)R were detected by real-time RT-PCR and Western blot in Calu 6, together with sst(5) and to a lesser extent sst(3) and D(4)R. BIM-23A387 and BIM-23A370 significantly inhibited growth of Calu-6, whereas IGF-IGFBP secretion or expression was unaffected, suggesting a direct inhibitory effect. The inhibition of cell growth, measured by both [(3)H]thymidine incorporation and cell count, was significantly lower when individual SSA and DA control peptides or subtype-specific SSA and DA were tested. BIM-23A370 was more potent than BIM 23A387 (P < 0.001). These findings show that SS/D chimeras can inhibit Calu-6 proliferation in an IGF-independent manner and suggest that this enhanced potency might be because of the induction of SSR/DR dimerization. The Calu-6 cell line, constitutively expressing SSR and DR, provides a suitable model to elucidate the mechanism of action of SSA and DA on regulation of cell growth and to characterize the interaction between SSR and DR. PMID- 16046459 TI - Transmitting tissue ECM distribution and composition, and pollen germinability in Sarcandra glabra and Chloranthus japonicus (Chloranthaceae). AB - BACKGROUND: and Aims Free-flowing surface exudates at the stigmatic (wet versus dry stigma) and adaxial epidermis at the site of angiospermy in carpels of Chloranthaceous species have been proposed to comprise a continuous extracellular matrix (ECM) operating in pollen tube transmission to the ovary. The aim of this research was to establish the spatial distribution and histo/immunochemical composition of the ECM involved in pollen tube growth in Sarcandra glabra and Chloranthus japonicus (Chloranthaceae). METHODS: Following confirmation of the pollen tube pathway, the histo/immunochemical make-up of the ECM was determined with histochemistry on fresh tissue to detect cuticle, esterase, proteins, pectins, and lipids and immunolocalization at the level of the TEM on sections from cryofixed/freeze-substituted tissue to detect molecules recognized by antibodies to homogalacturonans (JIM7, 5), arabinogalactan-proteins (JIM13) and cysteine-rich adhesion (SCA). KEY RESULTS: Pollen germinability is low in both species. When grains germinate, they do so on an ECM comprised of an esterase positive cuticle proper (dry versus wet stigma). Pollen tubes do not track the surface ECM of stigma or adaxial epidermal cells at the site of angiospermy. Instead, tubes grow between stigmatic cells and subsequently along the inner tangential walls of the stigmatic and adaxial carpel cells at the site of angiospermy. Pollen tubes enter the ovary locule at the base of the funiculus. The stigmatic ECM is distinct by virtue of the presence of anti-JIM5 aggregates, lipids, and a protein recognized by anti-SCA. CONCLUSIONS: The Chloranthaceae joins a growing number of basal angiosperm taxa whereby pollen tubes germinate on a dry versus wet stigma to subsequently grow intercellularly en route to the ovary thereby challenging traditional views that the archetype pollen tube pathway was composed of the surface of stigma and adaxial epidermal cells covered with a free-flowing exudate. PMID- 16046460 TI - Competition for nitrogen between Australian native grasses and the introduced weed Nassella trichotoma. AB - BACKGROUND: and Aims Nassella trichotoma is an unpalatable perennial grass weed that invades disturbed native grasslands in temperate regions of south-eastern Australia. This experiment investigated whether elevated N levels, often associated with disturbance, increases the competitiveness of N. trichotoma relative to C3 and C4 native Australian grasses. METHODS: A pot experiment investigated competitive interactions between four native grasses, two C3 species (Microlaena stipoides and Austrodanthonia racemosa) and two C4 species (Themeda australis and Bothriochloa macra), and N. trichotoma at three different N levels (equivalent to 0, 60 and 120 kg ha-1) and three competing densities (zero, one and eight neighbouring plants), using an additive design. KEY RESULTS: All native grasses were competitive with N. trichotoma at low N levels, but only M. stipoides was competitive at high N. High densities of native grasses (8:1) had a major competitive effect on N. trichotoma at all N levels. The competitive ranking of native grasses, across all N levels, on N. trichotoma was: M. stipoides>A. racemosa>B. macra>T. australis. The C3 species were generally more competitive than the C4 species and C4 grasses were not inherently more productive at low N levels, in contrast to the results of other studies. CONCLUSION: To resist invasion from N. trichotoma, these native grasses need to be maintained at a high density and/or biomass. The results do not support the theory that species such as N. trichotoma, with high tissues density, are always less competitive than those of low tissue density; in this case competitiveness depended on N levels. The ability of N. trichotoma to accumulate biomass at a higher rate than these native grasses, helps to explain why it is a major weed in disturbed Australian native grasslands. PMID- 16046461 TI - Methicillin-resistant Staphylococcus aureus heterogeneously resistant to vancomycin in a Turkish university hospital. AB - OBJECTIVES: We investigated vancomycin-intermediate Staphylococcus aureus (VISA) and heterogeneously vancomycin-intermediate S. aureus (hetero-VISA) isolates from clinical specimens of hospitalized patients at Hacettepe University over a 4 year period. METHODS: Strains were screened for VISA and hetero-VISA by using brain heart infusion agar containing 4 mg/L vancomycin (BHI-V4) and macro Etest. Confirmation of the isolates that were found to have intermediate susceptibility to vancomycin with either of the methods was done by population analysis of subpopulations with reduced susceptibility to vancomycin. The MIC of vancomycin for the isolates grown on BHI-V4 was determined by the microdilution method. RESULTS: Among 256 methicillin-resistant S. aureus (MRSA) isolates, 145 grew on BHI-V4. Forty-six of these were also found to be heterogeneously vancomycin intermediate strains when screened with the macro Etest. There were no VISA among 256 MRSA tested but 46 (17.97%) S. aureus strains with reduced susceptibility to vancomycin were identified by population analysis. Vancomycin MIC values for all isolates with reduced susceptibility were between or=1.0 mg/kg/day is as effective as amphotericin B at improving survival and reducing organ burdens in this murine model of disseminated C. tropicalis. PMID- 16046463 TI - Evaluation of a 10 {micro}g cefoxitin disc for the detection of methicillin resistance in Staphylococcus aureus by BSAC methodology. PMID- 16046464 TI - Surveillance and epidemiology of MRSA bacteraemia in the UK. AB - Surveillance of bacteraemia caused by methicillin-resistant Staphylococcus aureus (MRSA) in the UK has involved collection of data from hospital microbiology laboratories via several mechanisms, including a voluntary reporting scheme that has been operational in England and Wales since 1989 and mandatory reporting schemes that have been running independently in England, Wales, Scotland and Northern Ireland since 2001. In addition, surveillance schemes involving panels of participating sentinel laboratories that submit isolates for centralized susceptibility testing, such as the Bacteraemia Resistance Surveillance Programme run by the BSAC, have also been established. Each of these data sources have particular advantages, but they also have their individual limitations, with the result that they each give an incomplete picture if considered in isolation. However, by pooling the findings from these different but complementary surveillance programmes, a much more comprehensive and credible picture of the problem posed by MRSA is produced. These schemes have shown both a dramatic rise in the total numbers of cases of S. aureus bacteraemia reported annually and an increase in the proportion of such cases that involve MRSA (from 2% in 1990 to >40% in the early 2000s), although the most recent data indicate a slight reversal of these trends. Characterization of isolates of MRSA shows a marked temporal relationship between the rise in MRSA bacteraemias and the emergence and spread of two strains of epidemic MRSA, EMRSA-15 and EMRSA-16. Surveillance and control of MRSA infection continue to be high profile and further developments to the mandatory surveillance system in England are likely in the near future. PMID- 16046465 TI - Use of Etest MBL strips for the detection of carbapenemases in Acinetobacter baumannii. PMID- 16046466 TI - Molecular basis of resistance to trimethoprim, chloramphenicol and sulphonamides in Bordetella bronchiseptica. AB - OBJECTIVES: To date, little is known about the molecular basis of antimicrobial resistance in Bordetella bronchiseptica, an important respiratory tract pathogen in pigs, dogs and cats. The aim of this study was to identify genes coding for trimethoprim resistance present in porcine B. bronchiseptica and to determine their localization, transferability and association with other resistance genes. METHODS: Six B. bronchiseptica isolates with elevated MICs of trimethoprim were investigated by PCR for the presence of trimethoprim resistance genes and their association with class 1 integrons. The amplicons obtained were cloned and sequenced. Plasmid localization of these integrons was confirmed by transformation and conjugation. Isolates carrying the same integron were compared for their genetic relatedness by XbaI and SpeI pulsed-field gel electrophoresis (PFGE). RESULTS: Five B. bronchiseptica isolates carried a class 1 integron with two gene cassettes, one carrying the trimethoprim resistance gene dfrA1 and the other the chloramphenicol resistance gene catB3. This integron was present on a common conjugative plasmid in four of the five isolates and on the chromosome in the remaining isolate. All five B. bronchiseptica isolates proved to be related on the basis of their PFGE patterns. Another isolate had a class 1 integron with a dfrB1 and a catB2 cassette on a structurally different conjugative plasmid. The sulphonamide resistance gene sul1 was detected in the 3'-conserved segment of both types of integrons. CONCLUSIONS: This is the first report of trimethoprim, chloramphenicol and sulphonamide resistance genes and class 1 integrons in B. bronchiseptica isolates. PMID- 16046467 TI - Successful treatment of Acinetobacter meningitis with meropenem and rifampicin. PMID- 16046468 TI - In vitro and in vivo activity of combination antimicrobial agents on Haemophilus ducreyi. AB - OBJECTIVES: Development of single dose antibiotic treatments for chancroid has been followed by drug-resistant Haemophilus ducreyi in endemic areas. We examined the activity and interactions of antimicrobial agents and combinations against H. ducreyi. METHODS: We evaluated the in vitro susceptibility of three virulent strains of H. ducreyi to ceftriaxone, azithromycin, rifabutin and streptomycin, and each two-drug combination by the agar dilution method. We then tested each two-antibiotic combination for activity by the chequerboard method. Lastly, we chose the antibiotic combination with the lowest fractional inhibitory concentration index (FICI) and tested combined sub-therapeutic doses, the highest doses which had no effect alone on lesion healing compared with controls, for in vivo interaction in the temperature-dependent rabbit model of H. ducreyi infection. RESULTS: Each H. ducreyi strain was susceptible in vitro to each antibiotic and two-antibiotic combination, and combined ceftriaxone and streptomycin had the lowest FICI at 0.63. In five treated animals versus three untreated controls, combined sub-therapeutic doses of ceftriaxone (0.05 mg/kg) and streptomycin (10 mg/kg) reduced mean (SD) duration of culture positivity from 7.3 (1.1) to 2.6 (1.7) days (P<0.001), time to 50% reduction in lesion size from 9.7 (1.5) to 5.8 (0.8) days (P<0.005), and time to resolution of ulcer from 11.7 (2.3) to 6.6 (1.7) days (P<0.05). CONCLUSIONS: Ceftriaxone and streptomycin have in vivo synergic interaction against H. ducreyi lesions in the temperature dependent rabbit model of infection. Antibiotic combinations may be evaluated clinically as single-dose therapy for chancroid. PMID- 16046469 TI - Mechanisms of resistance to fluoroquinolones and carbapenems in Pseudomonas putida. AB - OBJECTIVES: Pseudomonas putida is an uncommon opportunistic pathogen, usually susceptible to antimicrobial agents. Data concerning resistance to antimicrobial agents in clinical P. putida isolates are limited. PATIENTS AND METHODS: Susceptibilities to fluoroquinolones, carbapenems and other antibiotics were characterized in five clinical isolates of P. putida recovered from different patients with urinary tract infections as causative pathogens. Fluoroquinolone and carbapenem resistance were characterized genetically by the methods of PCR and DNA sequencing. Outer membrane protein (OMP) profiles were characterized by SDS-PAGE. RESULTS: Four of five isolates were resistant or intermediate to both fluoroquinolones and carbapenems. Nucleotide sequences in the quinolone resistance-determining regions suggested that amino acid mutations such as Thr-83 ->Ile in GyrA and Glu-469-->Asp in GyrB may contribute to high resistance to fluoroquinolones. Four metallo-beta-lactamase-producing isolates that showed resistance to carbapenems carried the IMP-type metallo-beta-lactamase genes. A combined effect of reduced production of 46 kDa OMP and metallo-beta-lactamase production was shown by a P. putida isolate exhibiting the highest MICs of carbapenems. CONCLUSIONS: This study identified mechanisms of resistance to fluoroquinolones and carbapenems in clinical P. putida isolates. PMID- 16046470 TI - Type 4 sphingosine 1-phosphate G protein-coupled receptor (S1P4) transduces S1P effects on T cell proliferation and cytokine secretion without signaling migration. AB - Sphingosine 1-phosphate (S1P) has diverse effects on T cells that are mediated by the predominant S1P1 and S1P4 G protein-coupled receptors (GPCRs). S1P4 is expressed principally by leukocytes, but little is known of its T cell effects in immunity. Two approaches were used to investigate S1P4 signals in T cells. First, S1P4 was introduced into D10G4.1 mouse Th2 cells and EL4.IL-2 mouse T cells lacking endogenous S1P GPCRs. Second, mouse splenic CD4 T cells were treated with FTY720 to suppress S1P1 and leave S1P4 GPCRs as the only functionally relevant S1P receptor. Unlike S1P1, S1P4 failed to transduce chemotactic responses of any of the S1P4-only T cells to S1P or the phyto-S1P ligand selective for S1P4, or to suppress their chemotactic responses to chemokines. The S1P-S1P4 axis significantly inhibited T cell proliferation in each of the S1P4-only T cells activated by anti-CD3 and anti-CD28 MoAbs. Secretion of IL-4 by S1P4-D10G4.1 cells, IL-2 by S1P4-EL4.IL-2, and IFN-gamma by FTY720-treated CD4 T cells were significantly inhibited by S1P. In contrast, S1P enhanced secretion of IL-10 by stimulated S1P4-D10G4.1 T cells. Thus, S1P4 mediates immunosuppressive effects of S1P by inhibiting proliferation and secretion of effector cytokines, while enhancing secretion of the suppressive cytokine IL-10. PMID- 16046471 TI - IKKbeta phosphorylates p65 at S468 in transactivaton domain 2. AB - The transcription factor nuclear factor-kappa B (NF-kappaB) subunit p65 is phosphorylated by IkappaB kinase (IKK) at S536 in transactivation domain (TAD) 1. In this study, we investigate the presence of IKK sites in TAD2 of p65. Recombinant IKKbeta, but not IKKalpha, phosphorylated a GST-p65 substrate in which TAD1 was deleted. Mutational analysis revealed S468 as the only IKK site in TAD2. S468 phosphorylation occurred rapidly after TNF-alpha and IL-1beta in T cell, B cell, cervix carcinoma, hepatoma, breast cancer, and astrocytoma lines and in primary hepatic stellate cells as well as peripheral blood mononuclear cells. S468-phosphorylated p65 coimmunoprecipitated with IkappaBalpha, indicating that p65 is phosphorylated while bound to IkappaBalpha. Dominant negative IKKbeta or pharmacological IKK inhibition blocked S468 phosphorylation after TNF-alpha or IL-1beta, whereas dominant negative IKKalpha or inhibitors of MEK, p38, JNK, PI-3 kinase, or GSK-3 had no effect. p65S468A-reconstituted p65-/- mouse embryonic fibroblasts (MEFs) showed a small, but significant, elevation of NF-kappaB-driven luciferase activity and RANTES mRNA levels after TNF-alpha and IL-1beta in comparison to wtp65-reconstituted MEFs. p65 nuclear translocation was not altered in p65S468A-expressing MEFs. In conclusion, our results indicate that 1) IKKbeta phosphorylates multiple p65 sites, 2) IKKbeta phosphorylates p65 in an IkappaB p65 complex, and 3) S468 phosphorylation slightly reduces TNF-alpha- and IL-1beta induced NF-kappaB activation. PMID- 16046472 TI - Distinct molecular targets for the central respiratory and cardiac actions of the general anesthetics etomidate and propofol. AB - General anesthetics are among the most widely used and important therapeutic agents. The molecular targets mediating different endpoints of the anesthetic state in vivo are currently largely unknown. The analysis of mice carrying point mutations in neurotransmitter receptor subunits is a powerful tool to assess the contribution of the respective receptor subtype to the pharmacological actions of clinically used general anesthetics. We examined the involvement of beta3 containing GABA(A) receptors in the respiratory, cardiovascular, hypothermic, and sedative actions of etomidate and propofol using beta3(N265M) knock-in mice carrying etomidate- and propofol-insensitive beta3-containing GABA(A) receptors. Although the respiratory depressant action of etomidate and propofol, as determined by blood gas analysis, was almost absent in beta3(N265M) mice, the cardiac depressant and hypothermic effects, as determined by radiotelemetry, and the sedative effect, as determined by decrease of motor activity, were still present. Taken together with previous findings, our results show that both immobilization and respiratory depression are mediated by beta3-containing GABA(A) receptors, hypnosis by both beta3- and beta2-containing GABA(A) receptors, while the hypothermic, cardiac depressant, and sedative actions are largely independent of beta3-containing GABA(A) receptors. PMID- 16046473 TI - Changes in dysferlin, proteins from dystrophin glycoprotein complex, costameres, and cytoskeleton in human soleus and vastus lateralis muscles after a long-term bedrest with or without exercise. AB - This study was designed to evaluate the effects of hypokinesia and hypodynamia on cytoskeletal and related protein contents in human skeletal muscles. Twelve proteins: dystrophin and its associated proteins (DGC), dysferlin, talin, vinculin and meta-vinculin, alpha-actinin, desmin, actin, and myosin, were quantitatively analyzed during an 84-day long-term bedrest (LTBR). The preventive or compensatory effects of maximal resistance exercise (MRE) as a countermeasure were evaluated. Most of these proteins are involved in several myopathies, and they play an important role in muscle structure, fiber cohesion, cell integrity maintenance, and force transmission. This is the first comparison of the cytoskeletal protein contents between slow postural soleus (SOL) and mixed poly functional vastus lateralis (VL) human muscles. Protein contents were higher in VL than in SOL (from 12 to 94%). These differences could be mainly explained by the differential mechanical constraints imposed on the muscles, i.e., cytoskeletal protein contents increase with mechanical constraints. After LTBR, proteins belonging to the DGC, dysferlin, and proteins of the costamere exhibited large increases, higher in SOL (from 67 to 216%) than in VL (from 32 to 142%). Plasma membrane remodeling during muscle atrophy is probably one of the key points for interpreting these modifications, and mechanisms other than those involved in the resistance of the cytoskeleton to mechanical constraints may be implicated (membrane repair). MRE compensates the cytoskeletal changes induced by LTBR in SOL, except for gamma-sarcoglycan (+70%) and dysferlin (+108%). The exercise only partly compensated the DGC changes induced in VL, and, as for SOL, dysferlin remained largely increased (+132%). Moreover, vinculin and metavinculin, which exhibited no significant change in VL after LTBR, were increased with MRE during LTBR, reinforcing the pre-LTBR differences between SOL and VL. This knowledge will contribute to the development of efficient space flight countermeasures and rehabilitation methods in clinical situations where musculoskeletal unloading is a component. PMID- 16046474 TI - Hydrogen peroxide alters membrane and cytoskeleton properties and increases intercellular connections in astrocytes. AB - Excess hydrogen peroxide (H2O2) is produced in the pathogenesis of brain injuries and neurodegenerative diseases. H2O2 may damage cells through direct oxidation of lipids, proteins and DNA or it can act as a signaling molecule to trigger intracellular pathways leading to cell death. In this study, H2O2 caused plasma membranes of primary astrocytes to become more gel-like, while artificial membranes of vesicles composed of rat brain lipid extract became more liquid crystalline-like. Besides the effects on membrane phase properties, H2O2 promoted actin polymerization, induced the formation of cell-to-cell tunneling nanotube (TNT)-like connections among astrocytes and increased the colocalization of myosin Va with F-actin. Myosin Va was also observed in the H2O2-induced F-actin enriched TNT-like connections. Western blot analysis suggests that H2O2 triggered the phosphorylation of the p38 mitogen-activated protein kinase (MAPK), and that SB203580, a specific inhibitor of p38 MAPK, suppressed the changes in membrane phase properties and cytoskeleton resulting from H2O2 treatment. These results suggest that H2O2 alters astrocyte membranes and the cytoskeleton through activation of the p38 MAPK pathway. PMID- 16046475 TI - Drosophila Vps16A is required for trafficking to lysosomes and biogenesis of pigment granules. AB - Mutations that disrupt trafficking to lysosomes and lysosome-related organelles cause multiple diseases, including Hermansky-Pudlak syndrome. The Drosophila eye is a model system for analyzing such mutations. The eye-color genes carnation and deep orange encode two subunits of the Vps-C protein complex required for endosomal trafficking and pigment-granule biogenesis. Here we demonstrate that dVps16A (CG8454) encodes another Vps-C subunit. Biochemical experiments revealed a specific interaction between the dVps16A C-terminus and the Sec1/Munc18 homolog Carnation but not its closest homolog, dVps33B. Instead, dVps33B interacted with a related protein, dVps16B (CG18112). Deep orange bound both Vps16 homologs. Like a deep orange null mutation, eye-specific RNAi-induced knockdown of dVps16A inhibited lysosomal delivery of internalized ligands and interfered with biogenesis of pigment granules. Ubiquitous knockdown of dVps16A was lethal. Together, these findings demonstrate that Drosophila Vps16A is essential for lysosomal trafficking. Furthermore, metazoans have two types of Vps-C complexes with non-redundant functions. PMID- 16046476 TI - Inhibitory phosphorylation of a mitotic cyclin-dependent kinase regulates the morphogenesis, cell size and virulence of the smut fungus Ustilago maydis. AB - The regulation of cyclin-dependent kinase (CDK) activity through inhibitory phosphorylation seems to play an important role in the eukaryotic cell cycle. We have investigated the influence that inhibitory phosphorylation of the catalytic subunit of mitotic CDK has on cell growth and pathogenicity of the corn smut fungus Ustilago maydis. This model pathogen is worthy of attention since it is well suited to analyze the relationships between the cell cycle, morphogenesis and pathogenicity. We set out to study these relationships by producing a cdk1 mutant allele that was refractory to inhibitory phosphorylation. The expression of this mutant in U. maydis cells dramatically altered their morphology. Since this kind of mutation makes the CDK catalytic subunit resistant to regulation by Wee1-related kinases in other organisms, we characterized the orthologous Wee1 kinase from U. maydis. We found that Wee1 is essential in U. maydis. Overexpression of wee1 produces cell cycle arrest in G2, the target of Wee1 apparently being the Cdk1/Clb2 complex, which is required specifically for the onset of mitosis. Given the connection between the cell cycle control and pathogenesis in U. maydis, we also analyzed whether cells with impaired inhibitory phosphorylation of Cdk1 were able to infect plants. We found that inhibitory phosphorylation was required for mating, a prerequisite to initiate pathogenic development. By examining plant-specific expression of the constitutively unphosphorylated cdk1(AF) allele, we also found that appropriate levels of inhibitory phosphorylation were required at stages of infection subsequent to penetration by the fungus. These data reinforces the connections between cell cycle, morphogenesis and virulence in this smut fungus. PMID- 16046477 TI - Actin remodeling requires ERM function to facilitate AQP2 apical targeting. AB - This study provides the first evidence that actin reorganization during AQP2 vesicular trafficking to the plasma membrane requires the functional involvement of ERM (ezrin/radixin/moesin) proteins cross-linking actin filaments with plasma membrane proteins. We report that forskolin stimulation was associated with a redistribution of moesin from intracellular sites to the cell cortex and with a concomitant enrichment of moesin in the particulate fraction in renal cells. Introduction of a peptide reproducing a short sequence of moesin within the binding site for F-actin induced all the key effects of forskolin stimulation, including a decrease in F-actin, translocation of endogenous moesin, and AQP2 translocation. A straightforward explanation for these effects is the ability of the peptide to uncouple moesin from its putative effector. This modifies the balance between the active and inactive forms of moesin. Extraction with Triton X 100, which preserves cytoskeletal associated proteins, showed that forskolin stimulation or peptide introduction reduced the amount of phophorylated moesin, a molecular modification known to stabilize moesin in an active state. Our data point to a dual role of moesin in AQP2 trafficking: it might modulate actin depolymerization and it participates in the reorganization of F-actin-containing cytoskeletal structures close to the fusion sites of the AQP2-bearing vesicles. PMID- 16046478 TI - Induction of heat shock proteins in B-cell exosomes. AB - Exosomes are nanometer-sized vesicles secreted by a diverse range of live cells that probably have physiological roles in modulating cellular immunity. The extracellular factors that regulate the quantity and phenotype of exosomes produced are poorly understood, and the properties of exosomes that dictate their immune functions are not yet clear. We investigated the effect of cellular stress on the exosomes produced by B-lymphoblastoid cell lines. Under steady-state conditions, the exosomes were positive for hsp27, hsc70, hsp70 and hsp90, and other recognised exosome markers such as MHC class I, CD81, and LAMP-2. Exposing cells to heat stress (42 degrees C for up to 3 hours), resulted in a marked increase in these heat shock proteins (hsps), while the expression of other stress proteins such as hsp60 and gp96 remained negative, and other exosome markers remained unchanged. Stress also triggered a small increase in the quantity of exosomes produced [with a ratio of 1.245+/-0.07 to 1 (mean+/-s.e.m., n=20) of 3-hour-stress-exosomes to control-exosomes]. Flow-cytometric analysis of exosome-coated beads and immuno-precipitation of intact exosomes demonstrated that hsps were located within the exosome lumen, and not present at the exosome surface, suggesting that such exosomes may not interact with target cells through cell-surface hsp-receptors. Functional studies further supported this finding, in that exosomes from control or heat-stressed B cells did not trigger dendritic cell maturation, assessed by analysis of dendritic-cell-surface phenotype, and cytokine secretion profile. Our findings demonstrate that specific alterations in exosome phenotype are a hitherto unknown component of the cellular response to environmental stress and their extracellular function does not involve the direct activation of dendritic cells. PMID- 16046479 TI - MARCKS is a major PKC-dependent regulator of calmodulin targeting in smooth muscle. AB - Calmodulin (CaM) is a ubiquitous transducer of intracellular Ca(2+) signals and plays a key role in the regulation of the function of all cells. The interaction of CaM with a specific target is determined not only by the Ca(2+)-dependent affinity of calmodulin but also by the proximity to that target in the cellular environment. Although a few reports of stimulus-dependent nuclear targeting of CaM have appeared, the mechanisms by which CaM is targeted to non-nuclear sites are less clear. Here, we investigate the hypothesis that MARCKS is a regulator of the spatial distribution of CaM within the cytoplasm of differentiated smooth muscle cells. In overlay assays with portal-vein homogenates, CaM binds predominantly to the MARCKS-containing band. MARCKS is abundant in portal-vein smooth muscle ( approximately 16 microM) in comparison to total CaM ( approximately 40 microM). Confocal images indicate that calmodulin and MARCKS co distribute in unstimulated freshly dissociated smooth-muscle cells and are co targeted simultaneously to the cell interior upon depolarization. Protein-kinase C (PKC) activation triggers a translocation of CaM that precedes that of MARCKS and causes multisite, sequential MARCKS phosphorylation. MARCKS immunoprecipitates with CaM in a stimulus-dependent manner. A synthetic MARCKS effector domain (ED) peptide labelled with a photoaffinity probe cross-links CaM in smooth-muscle tissue in a stimulus-dependent manner. Both cross-linking and immunoprecipitation increase with increased Ca(2+) concentration, but decrease with PKC activation. Introduction of a nonphosphorylatable MARCKS decoy peptide blocks the PKC-mediated targeting of CaM. These results indicate that MARCKS is a significant, PKC-releasable reservoir of CaM in differentiated smooth muscle and that it contributes to CaM signalling by modulating the intracellular distribution of CaM. PMID- 16046480 TI - PI3K activation by IGF-1 is essential for the regulation of membrane expansion at the nerve growth cone. AB - Exocytotic incorporation of plasmalemmal precursor vesicles (PPVs) into the cell surface is necessary for axonal outgrowth and is known to occur mainly at the nerve growth cone. We have demonstrated recently that plasmalemmal expansion is regulated at the growth cone by IGF-1, but not by BDNF, in a manner that is quasi independent of the neuron's perikaryon. To begin elucidating the signaling pathway by which exocytosis of the plasmalemmal precursor is regulated, we studied activation of the IRS/PI3K/Akt pathway in isolated growth cones and hippocampal neurons in culture stimulated with IGF-1 or BDNF. Our results show that IGF-1, but not BDNF, significantly and rapidly stimulates IRS/PI3K/Akt and membrane expansion. Inhibition of PI3K with Wortmannin or LY294002 blocked IGF-1 stimulated plasmalemmal expansion at the growth cones of cultured neurons. Finally, our results show that, upon stimulation with IGF-1, most active PI3K becomes associated with distal microtubules in the proximal or central domain of the growth cone. Taken together, our results suggest a critical role for IGF-1 and the IRS/PI3K/Akt pathway in the process of membrane assembly at the axonal growth cone. PMID- 16046481 TI - Bub1 and aurora B cooperate to maintain BubR1-mediated inhibition of APC/CCdc20. AB - The spindle checkpoint maintains genome stability by inhibiting Cdc20-mediated activation of the anaphase promoting complex/cyclosome (APC/C) until all the chromosomes correctly align on the microtubule spindle apparatus via their kinetochores. BubR1, an essential component of this checkpoint, localises to kinetochores and its kinase activity is regulated by the kinesin-related motor protein Cenp-E. BubR1 also inhibits APC/C(Cdc20) in vitro, thus providing a molecular link between kinetochore-microtubule interactions and the proteolytic machinery that regulates mitotic progression. Several other protein kinases, including Bub1 and members of the Ipl1/aurora family, also regulate anaphase onset. However, in human somatic cells Bub1 and aurora B kinase activity do not appear to be essential for spindle checkpoint function. Specifically, when Bub1 is inhibited by RNA interference, or aurora kinase activity is inhibited with the small molecule ZM447439, cells arrest transiently in mitosis following exposure to spindle toxins that prevent microtubule polymerisation. Here, we show that mitotic arrest of Bub1-deficient cells is dependent on aurora kinase activity, and vice versa. We suggest therefore that the checkpoint is composed of two arms, one dependent on Bub1, the other on aurora B. Analysis of BubR1 complexes suggests that both of these arms converge on the mitotic checkpoint complex (MCC), which includes BubR1, Bub3, Mad2 and Cdc20. Although it is known that MCC components can bind and inhibit the APC/C, we show here for the first time that the binding of the MCC to the APC/C is dependent on an active checkpoint signal. Furthermore, we show that both Bub1 and aurora kinase activity are required to promote binding of the MCC to the APC/C. These observations provide a simple explanation of why BubR1 and Mad2 are essential for checkpoint function following spindle destruction, yet Bub1 and aurora B kinase activity are not. Taken together with other observations, we suggest that these two arms respond to different spindle cues: whereas the Bub1 arm monitors kinetochore-microtubule attachment, the aurora B arm monitors biorientation. This bifurcation in the signalling mechanism may help explain why many tumour cells mount a robust checkpoint response following spindle damage, despite exhibiting chromosome instability. PMID- 16046482 TI - Accelerated dendritic-cell migration and T-cell priming in SPARC-deficient mice. AB - On their path to draining lymph nodes, epidermal Langerhans cells traverse collagen-dense connective tissue before reaching lymphatic vessels. The matricellular protein SPARC (secreted protein, acidic and rich in cysteine), which is induced during inflammation and tissue repair, organizes collagen deposition in tissue stroma. We analyzed Langerhans cell and dendritic-cell migration and its impact on T-cell priming in SPARC-null (SPARC(-/-)) and SPARC sufficient (SPARC(+/+)) mice. Although the same number of Langerhans cells populate the ear skin of SPARC(-/-) and SPARC(+/+) mice, more Langerhans cells were found in the lymph nodes draining antigen-sensitized ears of SPARC(-/-) mice and significantly more Langerhans cells migrated from null-mice-derived ear skin explants. Such favored Langerhans cell migration is due to the host environment, as demonstrated by SPARC(+/+)>SPARC(-/-) and reciprocal chimeras, and have a profound influence on T-cell priming. Contact-, delayed type-hypersensitivity and naive T-cell receptor-transgenic T-cell priming, together indicate that the lack of SPARC in the environment accelerates the onset of T-cell priming by hastening Langerhans cell/dendritic-cell migration. PMID- 16046483 TI - Pathophysiology of mifepristone-induced septic shock due to Clostridium sordellii. AB - OBJECTIVE: To explain the role of mifepristone in medical abortions that results in fulminant and lethal septic shock due to Clostridium sordellii. DATA SOURCES: MEDLINE, PubMed, and Google Scholar databases were searched (1984-March 2005). Key search terms were mifepristone, RU38486, RU486, Mifeprex, medical abortion, septic shock, innate immune system, cytokines, and Clostridium sordellii. STUDY SELECTION AND DATA EXTRACTION: All articles identified from the data sources were evaluated and all information deemed relevant was included for the information related to the development of the understanding of the pathophysiology of mifepristone-induced septic shock due to C. sordellii. DATA SYNTHESIS: The mechanisms of action of mifepristone were incorporated into the pathophysiology of septic shock due to C. sordellii. Mifepristone, by blocking both progesterone and glucocorticoid receptors, interferes with the controlled release and functioning of cortisol and cytokines. Failure of physiologically controlled cortisol and cytokine responses results in an impaired innate immune system that results in disintegration of the body's defense system necessary to prevent the endometrial spread of C. sordellii infection. The abnormal cortisol and cytokine responses due to mifepristone coupled to the release of potent exotoxins and an endotoxin from C. sordellii are the major contributors to the rapid development of lethal septic shock. CONCLUSIONS: Theoretically, it appears that the mechanisms of mifepristone action favor the development of infection that leads to septic shock and intensifies the actions of multiple inflammatory cytokines, resulting in fulminant, lethal septic shock. PMID- 16046484 TI - The role of coenzyme Q10 in heart failure. AB - OBJECTIVE: To review the clinical data demonstrating the safety and efficacy of coenzyme Q10 (CoQ10) in heart failure (HF). DATA SOURCES: Pertinent literature was identified through MEDLINE (1966-January 2005) using the search terms coenzyme Q10, heart failure, antioxidants, and oxidative stress. Only articles written in the English language and evaluating human subjects were used. DATA SYNTHESIS: HF impairs the ability of the heart to maintain its normal cardiac output. Following an initial insult, cardiac remodeling ensues, resulting in left ventricular dilation and hypertrophy. Oxidative stress is also increased, while CoQ10 levels are decreased in patients with HF. This has led to the hypothesis that CoQ10, an antioxidant, may decrease oxidative stress, impair remodeling, and improve cardiac function. CONCLUSIONS: Large, well-designed studies on this topic are lacking. The limited data from well-designed trials indicate there may be some minor benefits with CoQ10 therapy in ejection fraction and end diastolic volume. CoQ10 therapy has been shown to be relatively safe with a low incidence of adverse effects. PMID- 16046485 TI - Amoxicillin/telithromycin-induced rash in infectious mononucleosis. PMID- 16046486 TI - Patient characteristics influencing evaluation of written medicine information: lessons for patient education. AB - BACKGROUND: Written medicine information (WMI) is considered an important component of patient education. Despite the wealth of information on many aspects of WMI, there is a paucity of studies examining how patient characteristics influence use and evaluation of WMI. OBJECTIVE: To investigate the influence of patient characteristics on the evaluation and intended future use of consumer medicine information (CMI), a form of WMI. METHODS: A questionnaire was administered to patients from 3 rheumatology/pain clinics in teaching hospitals and 40 community pharmacies. The questionnaire examined patients' perceptions of CMI (comprehension, perceived usefulness, design rating) and likelihood of using CMI in the future. Information on patient characteristics (demographic data, health literacy level) was also collected. Multiple regression analysis was used to examine associations between patient characteristics and their evaluation and intended future use of CMI. RESULTS: A total of 479 patients participated. Comprehension of CMI was associated with speaking primarily English at home, having attained secondary education or higher, and having adequate health literacy levels. Perceived usefulness of CMI was influenced by age and number of medications. Design rating was influenced by type of CMI, patient age, gender, and highest level of education. Intended future use was affected by health literacy level. In addition to individual patient characteristics, overall comprehension and perceived usefulness of CMI also influenced its intended future use. CONCLUSIONS: Patient characteristics were found to influence evaluation and intended future use of CMI. These findings should be taken into consideration in future research, development of WMI, and education of patients in everyday practice. PMID- 16046487 TI - Quetiapine cross-reactivity with plasma tricyclic antidepressant immunoassays. AB - BACKGROUND: Toxicology screens obtained on patients who have overdosed on drugs frequently include tricyclic antidepressants (TCAs) as part of the evaluation. Quetiapine is an antipsychotic agent with structural similarity to the TCAs. OBJECTIVE: To determine whether quetiapine may cross-react with plasma TCA immunoassays in vitro using commonly available autoanalyzers. METHODS: Quetiapine stock solution was added to 9 separate samples of pooled drug-free human plasma to produce concentrations ranging from 1 to 640 ng/mL that were verified by gas chromatography. No quetiapine metabolites were present. Each spiked plasma sample was tested in a blinded fashion using the Abbott Tricyclic Antidepressant TDx Assay on the TDxFLx autoanalyzer in 2 separate laboratories, the Syva Emit tox Serum Tricyclic Antidepressant Assay on the AU400 autoanalyzer and the S TAD Serum Tricyclic Antidepressant Screen on the ACA-Star 300 autoanalyzer. The TDx assay is quantitative, while Emit and S TAD are qualitative screening assays with a threshold of 300 ng/mL for TCA positivity. The outcome of interest was a positive TCA result. RESULTS: The quantitative assay showed concentration-related TCA cross-reactivity beginning at quetiapine concentrations of 5 ng/mL. The 640 ng/mL spiked sample produced TCA results of 379 and 385 ng/mL in labs 1 and 2, respectively. The qualitative assays were screened as TCA positive at quetiapine concentrations of 160 and 320 ng/mL for the S TAD and Emit assays, respectively. CONCLUSIONS: Quetiapine cross-reacts with quantitative and qualitative plasma TCA immunoassays in a concentration-dependent fashion. Therapeutic use or overdose of quetiapine may result in a false-positive TCA immunoassay result. PMID- 16046488 TI - Duloxetine associated with smoking cessation. PMID- 16046489 TI - Identifying and assisting low-literacy patients with medication use: a survey of community pharmacies. AB - BACKGROUND: Nearly one-half of adult Americans have limited functional literacy skills. Low patient literacy is associated with poor medication adherence and health outcomes. However, little is known about how pharmacies address literacy related needs among patrons. OBJECTIVE: To determine the frequency with which pharmacies identify and provide appropriate assistance to patients with limited literacy skills and provide specific recommendations to help improve pharmacists' recognition of low health literacy, as well as strategies to improve adherence in this population. METHODS: Through a telephone-based survey of Atlanta-area pharmacies, we obtained information on (1) whether the pharmacy attempted to identify patients with limited literacy skills, (2) what measures were taken by the pharmacy to optimize the health care of low-literacy patients, especially with regard to medication adherence, and (3) what services the pharmacy offered to improve adherence in general. RESULTS: The response rate among eligible pharmacies was 96.8% (N = 30). Only 2 (7%) pharmacies reported attempting to identify literacy-related needs among their patrons. One of these facilities provided additional verbal counseling to assist low-literacy patients, and the other pharmacy involved family members, provided verbal counseling, and had patients repeat instructions to confirm comprehension. Most pharmacies reported availability of adherence aids that could help low-literacy patients if such patients were identified and targeted to receive additional assistance. These included verbal and written counseling (offered at 73% of pharmacies), packaging or organizing aids (27%), refill services (17%), and graphic or multimedia aids (13%). CONCLUSIONS: Pharmacies infrequently attempt to identify and assist patients with limited literacy skills. PMID- 16046490 TI - Clozapine-induced hyperlipidemia resolved after switch to aripiprazole therapy. AB - OBJECTIVE: To report a case of severe clozapine-induced hypercholesterolemia and hypertriglyceridemia that resolved after therapy was switched to aripiprazole. CASE SUMMARY: A 42-year-old white man with schizoaffective disorder experienced new-onset hyperlipidemia with the addition of clozapine therapy. Despite treatment with various antihyperlipidemic agents, his total cholesterol level reached 477 mg/dL and his triglyceride level reached 4758 mg/dL. After a decrease in adherence with clozapine and subsequent deterioration, the patient was hospitalized and his antipsychotic therapy was switched to aripiprazole. The patient's lipid levels improved dramatically to the point that antihyperlipidemic treatment was discontinued. Due to lack of adequate symptomatic relief of psychiatric symptoms, the patient was ultimately switched back to clozapine therapy, at which time his lipid levels started to worsen again. DISCUSSION: There is a critical scarcity of data that relate to aripiprazole-induced lipid changes. Some studies have suggested that aripiprazole is not associated with the development of hyperlipidemia. Our case indicates that aripiprazole therapy may not have an adverse effect on lipid levels, even in patients who have a history of hyperlipidemia induced by another atypical antipsychotic. CONCLUSIONS: Should aripiprazole be found to have a definitive lipid-neutral effect, then clinicians would be wise to factor this finding into overall benefit-versus-risk considerations in the antipsychotic treatment selection process, especially in a society in which cardiovascular disease continues to be a principal cause of morbidity and mortality. PMID- 16046491 TI - Impact of the clinical pharmacist on readmission in patients with acute coronary syndrome. AB - BACKGROUND: Previous studies have reported a positive impact of pharmacists on care of patients with chronic illnesses. The impact of the clinical pharmacist on hospital readmission in patients with acute coronary syndromes (ACS) has yet to be evaluated, as of this writing. OBJECTIVE: To evaluate the impact of the clinical pharmacist as a direct patient-care team member on cardiac-related readmission in patients admitted to the general cardiology unit with ACS. METHODS: A prospective, nonrandomized observational study compared patients who received standard practice care with patients admitted to a service with a clinical pharmacist to provide care at the bedside. Patients admitted to and discharged from the general cardiology unit for ACS were included. The primary endpoint of the study was cardiac-related readmission at 30 days following hospital discharge. Secondary endpoints included length of stay and medication utilization. Interventions provided by the clinical pharmacist in the study group were documented. RESULTS: Cardiac readmission at 30 days was similar between the groups (p = 0.59). In the subset of patients with unstable angina, readmission in the study group was significantly lower than in the control group (1.3% vs 9.1%; p = 0.04). Patients in both groups were similarly managed using drug therapy and invasive coronary interventions. The medical staff's rate of acceptance of recommendations provided by the pharmacist was 94.4%. The most common interventions were medication education and identification of indicated therapy. CONCLUSIONS: The addition of pharmacists did not decrease readmission in patients with ACS. The finding of significant reduction in readmission in the subset of patients with unstable angina should be considered "hypothesis generating" for future randomized studies to confirm the results. PMID- 16046492 TI - BlastXtract--a new way of exploring translated searches. AB - SUMMARY: Searches of translated, unannotated genomic DNA sequences against protein databases is a useful early-stage method for discovering protein homologues encoded by the sequence, but generates huge amounts of output data that quickly become impregnable. BlastXtract is a web-based tool for managing and visualizing results from large translated BLAST and FastA searches. It combines the speed and storage benefits of relational database management systems with an easy-to-use graphical navigation map, and greatly facilitates the early exploration of genomic sequence. AVAILABILITY: BlastXtract can be downloaded from http://bioinfo.ucc.ie/blastxtract/. PMID- 16046493 TI - Extraction of regulatory gene/protein networks from Medline. AB - MOTIVATION: We have previously developed a rule-based approach for extracting information on the regulation of gene expression in yeast. The biomedical literature, however, contains information on several other equally important regulatory mechanisms, in particular phosphorylation, which we now expanded for our rule-based system also to extract. RESULTS: This paper presents new results for extraction of relational information from biomedical text. We have improved our system, STRING-IE, to capture both new types of linguistic constructs as well as new types of biological information [i.e. (de-)phosphorylation]. The precision remains stable with a slight increase in recall. From almost one million PubMed abstracts related to four model organisms, we manage to extract regulatory networks and binary phosphorylations comprising 3,319 relation chunks. The accuracy is 83-90% and 86-95% for gene expression and (de-)phosphorylation relations, respectively. To achieve this, we made use of an organism-specific resource of gene/protein names considerably larger than those used in most other biology related information extraction approaches. These names were included in the lexicon when retraining the part-of-speech (POS) tagger on the GENIA corpus. For the domain in question, an accuracy of 96.4% was attained on POS tags. It should be noted that the rules were developed for yeast and successfully applied to both abstracts and full-text articles related to other organisms with comparable accuracy. AVAILABILITY: The revised GENIA corpus, the POS tagger, the extraction rules and the full sets of extracted relations are available from http://www.bork.embl.de/Docu/STRING-IE PMID- 16046494 TI - CHORAL: a differential geometry approach to the prediction of the cores of protein structures. AB - MOTIVATION: Although the cores of homologous proteins are relatively well conserved, amino acid substitutions lead to significant differences in the structures of divergent superfamilies. Thus, the classification of amino acid sequence patterns and the selection of appropriate fragments of the protein cores of homologues of known structure are important for accurate comparative modelling. RESULTS: CHORAL utilizes a knowledge-based method comprising an amalgam of differential geometry and pattern recognition algorithms to identify conserved structural patterns in homologous protein families. Propensity tables are used to classify and to select patterns that most likely represent the structure of the core for a target protein. In our benchmark, CHORAL demonstrates a performance equivalent to that of MODELLER. PMID- 16046495 TI - pIQPNNI: parallel reconstruction of large maximum likelihood phylogenies. AB - SUMMARY: IQPNNI is a program to infer maximum-likelihood phylogenetic trees from DNA or protein data with a large number of sequences. We present an improved and MPI-parallel implementation showing very good scaling and speed-up behavior. PMID- 16046496 TI - TileMap: create chromosomal map of tiling array hybridizations. AB - MOTIVATION: Tiling array is a new type of microarray that can be used to survey genomic transcriptional activities and transcription factor binding sites at high resolution. The goal of this paper is to develop effective statistical tools to identify genomic loci that show transcriptional or protein binding patterns of interest. RESULTS: A two-step approach is proposed and is implemented in TileMap. In the first step, a test-statistic is computed for each probe based on a hierarchical empirical Bayes model. In the second step, the test-statistics of probes within a genomic region are used to infer whether the region is of interest or not. Hierarchical empirical Bayes model shrinks variance estimates and increases sensitivity of the analysis. It allows complex multiple sample comparisons that are essential for the study of temporal and spatial patterns of hybridization across different experimental conditions. Neighboring probes are combined through a moving average method (MA) or a hidden Markov model (HMM). Unbalanced mixture subtraction is proposed to provide approximate estimates of false discovery rate for MA and model parameters for HMM. AVAILABILITY: TileMap is freely available at http://biogibbs.stanford.edu/~jihk/TileMap/index.htm. SUPPLEMENTARY INFORMATION: http://biogibbs.stanford.edu/~jihk/TileMap/index.htm (includes coloured versions of all figures). PMID- 16046497 TI - Automated Microarray Image Analysis Toolbox for MATLAB. AB - The Automated Microarray Image Analysis (AMIA) Toolbox for MATLAB is a flexible, open-source, microarray image analysis tool that allows the user to customize analyses of microarray image sets. This tool provides several methods to identify and quantify spot statistics, as well as extensive diagnostic statistics and images to evaluate data quality and array processing. The open, modular nature of AMIA provides access to implementation details and encourages modification and extension of AMIA's capabilities. AVAILABILITY: The AMIA Toolbox is freely available at http://www.pnl.gov/statistics/amia. The AMIA Toolbox requires MATLAB 6.5 (R13) (MathWorks, Inc. Natick, MA), as well as the Statistics Toolbox 4.1 and Image Processing Toolbox 4.1 for MATLAB or more recent versions. CONTACT: amanda.white@pnl.gov PMID- 16046498 TI - A statistical model for HIV-1 sequence classification using the subtype analyser (STAR). AB - MOTIVATION: HIV-1 antiretroviral drug resistance testing produces large amounts of HIV-1 protease and reverse transcriptase sequences. These provide an excellent resource to study the incidence, spread and clinical significance of HIV-1 subtypes. We have produced a program, Subtype Analyser (STAR) that rapidly and accurately subtypes HIV-1. Here we have determined a robust and statistically validated model for subtype assignment. RESULTS: We have significantly extended our HIV-1 subtyping tool (STAR), such that each query sequence when evaluated against subtype profile alignments, returns a discriminating score based on the ratio of subtype positive to negative amino acid positions. These scores were transformed into a Z-score distribution and evaluated. Of the 141 sequences used to define the subtype alignments, 98% were correctly reclassified. Inclusion of additional recombination detection within STAR increased the detection of known recombinant sequences to 95%. AVAILABILITY: STAR is available as compiled (Linux Fedora 3) or source code from http://pgv19.virol.ucl.ac.uk/download/star_linux.tar CONTACT: p.kellam@ucl.ac.uk SUPPLEMENTARY INFORMATION: http://pgv19.virol.ucl.ac.uk/download/star_supplement PMID- 16046499 TI - Effect of radio contrast media on residual renal function in peritoneal dialysis patients--a prospective study. AB - BACKGROUND: Residual renal function is an independent predictor of survival in peritoneal dialysis patients. Systemic administration of radio contrast media (CM) may increase the risk of acute renal failure in patients with impaired renal function not on dialysis. There are few data on the influence of CM administration in dialysis patients. METHODS: We investigated residual renal function in 10 continuous ambulatory peritoneal dialysis (CAPD) patients who underwent elective diagnostic intravenous or intra-arterial administration of CM (study group). Iopromide (a iodinated, non-ionic hypo-osmolar CM) was used for all interventions. The median dose of CM given was 107.5 ml/patient. Residual renal function (calculated as the average of renal creatinine and renal urea clearance) was measured on the day before the intervention (baseline), on days 1 7, day 10 and day 30 after intervention. Eight CAPD patients without exposure to CM acted as the control group. RESULTS: There was no significant difference between the two groups in age, gender, diabetes, duration of dialysis and renal clearance at baseline. In the study group, we observed a temporary decline of residual renal clearance after administration of CM (P<0.05; Friedman test). On day 30, clearances were not significantly different from baseline. In the control group, there was no significant change of residual clearance during the observation period. Repeated measures ANOVA revealed no significant difference in the course of residual renal function between study and control groups. The decline of residual renal clearance between baseline and a routine visit after 4 months was comparable between groups. CONCLUSION: Administration of iopromide did not lead to a persistent decline of residual renal function in CAPD patients. Nevertheless, non-ionic hypo-osmolar CM should be given to these patients with the lowest possible dose and only if there is a real clinical indication. PMID- 16046500 TI - The relationship of plasma ghrelin level to energy regulation, feeding and left ventricular function in non-diabetic haemodialysis patients. AB - OBJECTIVE: Inadequate plasma ghrelin levels determine the suppression of appetite, nutritional state and morbidity. We investigated the correlation between plasma ghrelin levels and appetite, nutritional status and cardiovascular morbidity in maintenance haemodialysis (HD) patients. METHODS: We measured plasma ghrelin levels at 2 h intervals during a 24 h period in 22 non-diabetic HD patients, who were grouped into normal intake or disturbed intake according to subjective global assessments, and in six healthy controls. RESULTS: A significant positive correlation existed between the 24 h plasma ghrelin profile and most time-specific plasma ghrelin levels in non-diabetic HD patients. Ghrelin levels in the abnormal intake group were higher than those in the normal intake group. A significant positive correlation existed between ghrelin and left ventricle functions, including left ventricle mass index (r = 0.75, P = 0.008), left ventricle mass (r = 0.57, P = 0.03) and interventricular septum thickness (r = 0.70, P = 0.009). An inverse correlation existed between plasma ghrelin and right ventricular dimension (r = -0.45, P = 0.035). Body mass index (r = -0.64, P = 0.033) and body fat content (r = -0.619, P = 0.002) had significant negative correlations with plasma ghrelin. CONCLUSIONS: Anorexia was found in patients with higher plasma ghrelin levels. Plasma ghrelin levels in non-diabetic HD patients showed a significant correlation with left ventricular function. PMID- 16046501 TI - Fistula dysfunction secondary to a subcutaneous myelomatous deposit. PMID- 16046502 TI - Is there a link between cytomegalovirus infection and new-onset posttransplantation diabetes mellitus? Potential mechanisms of virus induced beta cell damage. PMID- 16046503 TI - Haemodialysis catheter-associated infection: common pathogens in unusual places. PMID- 16046504 TI - VEGF -460 genotype plays an important role in progression to chronic kidney disease stage 5. AB - BACKGROUND: Changes in renal vasculature, with vascular and interstitial fibrosis, are hallmarks of progression to chronic kidney disease (CKD) stage 5. Vascular endothelial growth factor (VEGF) is a potent angiogenic and vascular permeability factor. Transforming growth factor-beta1 (TGF-beta1) plays a critical role in promoting extracellular matrix (ECM) deposition and fibrosis. This study investigates whether genetic polymorphisms of VEGF or TGF-beta1 are associated with (i) progressive decline in renal function in patients with glomerular disorders (cohort 1) and (ii) predisposition to CKD stage 5 in a separate group of renal transplant recipients with various primary diseases (cohort 2). METHODS: Two patient groups were studied. Cohort 1 comprised 91 patients with biopsy-proven glomerular disease who were followed-up for 5 years before categorization as either non-progressors (with stable serum creatinine or < or =30% increase over 5 years, n = 39) or progressors (requiring dialysis, transplantation or whose serum creatinine increased by >30% over 5 years, n = 52). Cohort 2 comprised 107 patients with various primary renal diseases, who had reached CKD stage 5 and undergone renal transplantation at the time of study. All patients were genotyped for the VEGF polymorphisms at positions -460 (C/T) and +405 (G/C). Linkage disequilibrium (LD) was established using EHplus. SNPHAP was used to estimate haplotype frequency and to infer haplotypes to all patients. Cohort 1 patients were genotyped for the TGF-beta1 polymorphisms at positions 800, -509, codons 10 and 25. Genotyping was performed by polymerase chain reaction-restriction length polymorphism (PCR-RFLP). RESULTS: In cohort 1, there was a significant increase in frequency of the -460 VEGF CC genotype 30.8 vs 5.1%, P = 0.008; odds ratio (OR), CC vs TT 10.67, 95% confidence interval (CI), 1.94-58.72 and C allele 56.7 vs 37.2%, P = 0.009; OR 2.22, 95% CI, 1.21-4.04, in the progressor patients when compared with the non-progressors. In cohort 2, there was a significant increase in the VEGF -460 CC genotype when compared with healthy volunteers 37 vs 20.8%, P = 0.011; OR CC vs TT 1.59, 95% CI, 0.72-3.51. The -460 and +405 polymorphisms were in LD P < 0.00007. There were significant differences in diplotype (haplotype pair) frequencies in cohort 1 and 2, P = 0.018, which confirmed the importance of the -460C allele. There were no associations between the VEGF +405 or TGF-beta1 polymorphisms and progressive renal disease. CONCLUSION: In this study, we have demonstrated an association between the VEGF -460 polymorphism and progression to CKD stage 5. The function of this polymorphism remains unclear although previous evidence suggests that promoter constructs containing this single nucleotide polymorphism (SNP) have been associated with increased activity. Clearly there is a role for TGF-beta1 in chronic kidney disease. However, this study found no associations with four TGF beta1 polymorphisms in this cohort. PMID- 16046505 TI - Fibromyalgia: its prevalence in haemodialysis patients and its relationships with clinical and laboratory parameters. AB - OBJECTIVE: Our aim was to determine the prevalence of fibromyalgia syndrome (FS) in chronic haemodialysis (HD) patients and to identify possible links between FS and various clinical and laboratory parameters. METHODS: We studied 122 chronic HD patients and 89 healthy age- and sex-matched controls, classified according to the American College of Rheumatology (ACR) classification criteria for FS. Age, sex, causes of renal failure, length of time on dialysis and marital status were recorded, and questions were asked about symptoms related to FS. All subjects completed the Fibromyalgia Impact Questionnaire (FIQ). Laboratory data obtained in the preceding 6 months were re-evaluated. RESULTS: Nine (7.4%) of the 122 HD patients and four of the 89 control subjects (4.5%) fulfilled the ACR criteria for definite FS (P = 0.56). The mean ages of the subjects who had definite FS and those who did not were similar. Most of the subjects diagnosed with definite FS were female (11 out of 13). The HD patients had higher FIQ scores than the controls, regardless of FS diagnosis. Among the HD patients, those with definite FS had a significantly higher mean FIQ score than all the other HD patients combined. In the all HD patients group, fatigue, irritable bowel syndrome and personal histories of depression were correlated with FS, whereas duration of HD, aetiology of renal failure, laboratory parameters and hepatitis B or C virus infection were not. CONCLUSION: The prevalence of FS appeared to be similar in chronic HD patients and the general population; also, FS-related symptoms appear to be similar in HD patients and the general population who have FS. No laboratory parameter was correlated with frequency of FS. PMID- 16046506 TI - Effectiveness of a multidisciplinary kidney disease clinic in achieving treatment guideline targets. AB - BACKGROUND: We have demonstrated previously that at referral most chronic kidney disease (CKD) patients have suboptimal metabolic and hypertension control. Although several studies suggest that CKD clinics improve patient outcome, in fact there are minimal published data describing the actual effect of such clinics on these parameters. METHODS: We performed a historical prospective review of a cohort of 340 CKD patients referred to our multidsciplinary clinic in 1998 or 1999, with estimated creatinine clearance (CCr) <60 ml/min. Data regarding blood pressure (BP) control, metabolic/anaemia parameters, medications, access planning and dialysis starts were collected. RESULTS: The number of patients followed was 234, 144, 100 and 70 at years 1-4 of follow-up, respectively. Twenty-five percent of the patients were diabetic, and 25% were suspected to have ischaemic nephropathy; mean age was 67+/-15 years. Although phosphate control improved from referral, below a CCr of 30 ml/min, 27% of visits showed hyperphosphataemia. Thirty-one percent of patients with CCr <15 ml/min had haemoglobin <100 g/l at follow-up despite the availability of erythropoietin. BP improved from a mean of 151/80 mmHg at referral to 137/75 mmHg in subsequent visits. At follow-up visits, 62% of BPs were still >130 mmHg systolic or 85 mmHg diastolic. For proteinuric patients (>1 g/day), 75% of follow-up visits showed BP >125/75 mmHg, despite angiotensin-converting enzyme inhibitor use increasing from 35% at referral to 79% at follow-up. Twenty-four percent of patients started renal replacement therapy, initially haemodialysis (HD) in 57%, peritoneal dialysis (PD) in 35% and pre-emptive transplant in 8%. Thirty-eight percent of dialysis starts occurred within 6 months of referral, but PD was the modality in half of these. Only half of the HD patients started using an aterio-venous fistula, and of those using a central catheter 11 of 24 had been followed >6 months, but only four of them had attempted fistula creation. CONCLUSIONS: CKD clinic attendance was associated with improvements in metabolic and BP control, and was able to facilitate the use of PD even for late referrals. However, even the multidisciplinary model with nephrologists, nurse educators and dietitians was unable to achieve guideline-recommended metabolic, anaemia, BP and access targets for a significant number of patients. PMID- 16046507 TI - Effects of smoking on renal function in patients with type 1 and type 2 diabetes mellitus. AB - BACKGROUND: Smoking increases the risk of end-stage renal failure in patients with primary renal disease. Whether and to what extent smoking affects the kidneys in diabetic patients with normal renal function and variable degrees of proteinuria has not been fully studied. METHODS: We followed 185 patients with type 1 or 2 diabetes mellitus and with or without signs of overt renal disease for at least 3 years, median 5.1 (3-6.8) years. Each patient had a baseline visit and at least four follow-up visits (average 4.8+/-0.3). Cases were patients who were smoking (n = 44) at the time the survey was started. Controls were patients who had never smoked (n = 141). Glomerular filtration rate (GFR) was estimated using the MDRD formula. Multiple logistic regression was used to correct for confounding factors. RESULTS: At baseline, smokers were younger (47+/-14 vs 54+/ 16 years, P < 0.01), and had a lower GFR (95+/-26 ml/min) than non-smokers (107+/ 33 ml/min, P < 0.05). Mean GFR remained constant during follow-up in non-smokers (106+/-31 ml/min), but decreased significantly in smokers (83+/-22 ml/min, P < 0.0001), and this relationship persisted when adjusted for retinopathy, glycaemic control, age, body habitus, ACE-inhibitor treatment, blood pressure control or severity of proteinuria. The effect of smoking on GFR decline was stronger in patients with type 1 diabetes or male gender. CONCLUSIONS: Cigarette smoking causes a decrease in GFR in diabetic patients with normal or near-normal renal function, independent of confounding factors including severity of proteinuria. The latter finding suggests a mechanism independent of glomerular damage. PMID- 16046508 TI - Assessing the utility of the stop dialysate flow method in patients receiving haemodiafiltration. AB - BACKGROUND: The stop dialysate flow (SDF) method of post-dialysis urea sampling is the most commonly used method in the UK. It can also be used with a published formula to predict 30 min equilibrated urea accurately. The method has not been validated in patients undergoing haemodiafiltration (HDF). Given the increased use of HDF across Europe, we felt it prudent to assess the utility of the SDF method and prediction equation in this modality. METHODS: Fourteen patients from two renal units were studied. Blood samples were taken at 1 min intervals from the arterial side of the dialysis circuit in the first 5 min after HDF had ceased whilst blood circulation continued. A peripheral sample was taken from the contralateral arm immediately after HDF had ceased and a 30 min sample was taken from the arterial needle. These samples were used to assess the utility of 5 min arterial blood urea and the 30 min prediction formula, respectively. RESULTS: Blood urea measured from the arterial circuit at 5 min correlated closely with the contralateral sample taken immediately post-HDF, with no significant difference (6.45+/-2.11 vs 6.52+/-2.19 mmol/l, P = 0.39). The use of 5 min arterial blood urea and prediction formula allowed an accurate prediction of 30 min urea (R2 = 0.96). CONCLUSIONS: The use of the SDF method with a 5 min post HDF arterial sample is valid in patients receiving HDF. The previously published prediction formula for estimating 30 min urea is also valid using the 5 min post HDF sample. PMID- 16046509 TI - Drug interaction between sevelamer and furosemide. PMID- 16046510 TI - Successful reintroduction of a different erythropoiesis-stimulating agent after pure red cell aplasia: relapse after successful therapy with prednisone. AB - We report a 3-year case history that describes a 78-year-old woman with recurrent transfusion-dependent pure red cell aplasia (PRCA) secondary to recombinant epoetin use that was responsive to immunosuppressant therapy. The patient had kidney disease of unknown aetiology (estimated glomerular filtration rate of 13 ml/min/1.73 m2) and was not on dialysis. After 16 months of therapy with subcutaneous Eprex, she developed anti-erythropoietin antibody-confirmed PRCA and was started on high dose prednisone (50 mg per day). Within 5 months, the patient's serum was clear of antibodies and, under the cover of low dose prednisone (5-7.5 mg per day), therapy with a different erythropoiesis stimulating compound (Aranesp) was initiated due to persistent fatigue and anaemia. At 3 months of therapy, the serum anti-erythropoietin antibodies remained negative and, due to the patient's requests, and after discussion, prednisone therapy was discontinued. Unfortunately, 3 months after cessation of prednisone, a recurrence of PRCA was confirmed by the development of profound anaemia and reappearance of anti-erythropoietin antibodies in the patient's serum. High dose prednisone (50 mg per day) was reinstituted, whereupon, 2 months later, antibodies were again confirmed to be negative. This case report demonstrates the responsiveness of PRCA to simple immunosuppressive therapy, and the ability to introduce different erythropoiesis-stimulating agents in the presence of such therapy. It appears that there may be problems associated with discontinuation of immunosuppressive therapy in the presence of sustained erythropoiesis-stimulating agent therapy in those in whom the condition has occurred previously. PMID- 16046511 TI - A rare case of alveolar haemorrhage due to malignant hypertension. PMID- 16046512 TI - Melagatran anticoagulation during haemodialysis--'Primum non nocere'. PMID- 16046513 TI - Thalidomide for the nephrologist. PMID- 16046514 TI - A case of leptospirosis presenting with end-stage renal failure. PMID- 16046515 TI - Expression of matrix metalloproteinase-9 associated with ets-1 proto-oncogene in rat tubulointerstitial cells. AB - BACKGROUND: Ets-1 proto-oncogene exhibits multiple activities in the transcriptional regulation of numerous genes including metalloproteinase (MMP)-1, -3 and -9. MMPs play an important role in the remodelling of extracellular matrix in various renal diseases. However, the role of the Ets-1-MMP axis in advanced renal diseases is uncertain. In the present study, we investigated whether Ets-1 is involved in interleukin (IL)-1-mediated expression of MMPs in tubulointerstitial cells. METHODS: Rat renal fibroblasts (NRK-49F) and tubular epithelial cells (NRK-52E) were cultured and allocated to an IL-1beta-treated group (10 ng/ml), a platelet-derived growth factor (PDGF)-BB-treated group (25 ng/ml) and a control group. Protein and mRNA were extracted after 1, 6, 12 and 24 h of treatment. Parallel flasks were treated with 2 muM ets-1 antisense oligodeoxynucleotides (ODNs) before exposure to IL-1beta. The expression of Ets-1 protein was evaluated by western blotting. The activities of MMPs were evaluated by gelatin zymography. The expression of ets-1 and/or MMP-9 mRNA was evaluated semiquantitatively by real-time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: In NRK-49F cells, Ets-1 protein increased significantly by 6.8 fold at 6 h, and MMP-9 activity increased significantly by 9.9-fold at 12 h in the IL-1beta-treated group compared with controls. MMP-2 and -3 activities also increased significantly in the IL-1beta-treated group. In NRK-52E cells, Ets-1 protein was 3.1 times higher at 1 h, and the latent form of MMP-9 activity increased 3.4-fold at 6 h in the IL-1beta group compared with controls. However, MMP-2 or MMP-3 activities were not markedly altered by IL-1beta treatment compared with controls. When the cells were treated with ets-1 antisense ODNs before IL-1beta treatment, Ets-1 protein expression decreased at least 50%, and MMP-9 activity was clearly inhibited in both cells. We also confirmed that MMP-9 activity was upregulated on days 21 and 28 in renal cortex of rat crescentic glomerulonephritis. CONCLUSIONS: The Ets-1 transcriptional factor may participate in IL-1beta-mediated MMP-9 expression in tubulointerstitial cells. PMID- 16046516 TI - The impact of topical mupirocin on peritoneal dialysis infection in Singapore General Hospital. AB - BACKGROUND: Peritonitis and exit-site infections (ESI) are major causes of morbidity in peritoneal dialysis (PD) patients. The application of topical mupirocin to exit sites reduces such complications, and prolongs life in PD. Since the year 2000, this topical treatment has been used in our hospital on new PD patients. We analysed the results of this protocol, and studied the effects of comorbidities on the incidence of peritonitis. METHODS: We studied 740 incident PD patients, who were divided into two groups based on year of entry into PD (Group 1 from January 1998 to December 1999 inclusive, topical mupirocin not used, and Group 2 from January 2000 to March 2004 inclusive, topical mupirocin used). The variables we studied included gender, age, diabetic status, ischaemic heart disease, peripheral vascular disease, cerebrovascular disease and serum albumin. RESULTS: The application of topical mupirocin at the exit site led to a significant reduction in the rate of peritonitis (0.443 vs 0.339 episodes per patient-year; P<0.0005) and in ESI (0.168 vs 0.156 episodes per patient-year; P<0.005), results attributed primarily by the significant (P<0.005) reduction in Staphylococcus aureus infection. There was also an unexpected lowering of Pseudomonas aeruginosa peritonitis in the mupirocin group (P<0.005). Stepwise multiple logistic regression analysis revealed that only the application of mupirocin and serum albumin levels were significant predictors of peritonitis. CONCLUSIONS: Our study, although retrospective, has demonstrated that the topical use of mupirocin was associated with a significant reduction in ESI and peritonitis and, unexpectedly, with findings of fewer incidences of Pseudomonas peritonitis. Serum albumin level before the initiation of PD was a strong predictor of subsequent peritonitis. Mupirocin, with its low toxicity, ease of application and demonstrable beneficial effect in reducing ESI and peritonitis is now used on all of our incident PD patients. PMID- 16046517 TI - Questionnaire-based evaluation of gastrointestinal disorders in de novo renal transplant patients receiving either mycophenolate mofetil or enteric-coated mycophenolate sodium. AB - BACKGROUND: Gastrointestinal (GI) disorders are one of the main adverse events in patients treated by mycophenolic acid (MPA). The aim of our prospective questionnaire-based study was to assess GI side-effects in de novo renal transplant patients receiving either mycophenolate mofetil (MMF) or enteric coated mycophenolate sodium (EC-MPS). METHODS: Between January 2002 and April 2003, all patients receiving MPA with a functioning allograft at 1 month post transplantation were enrolled in this study (n = 130). Ninety-three of them received MMF (group I), and 37 patients received EC-MPS (group II). Each month, every patient completed a questionnaire regarding GI disorders. RESULTS: During the first year post-transplantation, GI disorders occurred in 31 patients from the MMF group (33.3%) and 12 patients from the EC-MPS group (32.4%) (not significant). The incidence of upper GI disorders was also similar in both groups. Diarrhoea was observed in 18 patients (19.3%) from group I, and in five patients from group II (13.5%) (not significant). Its frequency and severity were similar in both groups. Weight loss was observed in three patients receiving MMF. Diarrhoea resolved spontaneously in 10 patients from group I and in all patients from group II. For the other eight patients in group I, the diarrhoea required MMF discontinuation in three patients and dose reduction in five patients. CONCLUSIONS: In conclusion, in this questionnaire-based evaluation, the incidence of GI disorders was similar in patients receiving either MMF or EC-MPS during the first year post-transplantation. PMID- 16046518 TI - Herbal teas interfere with cyclosporin levels in renal transplant patients. PMID- 16046519 TI - Age, the riddle of renal transplantation. PMID- 16046520 TI - Health status and quality of life reported by incident patients after 1 year on haemodialysis or peritoneal dialysis. AB - BACKGROUND: It has been suggested that there are no large differences in the quality of life of incident patients starting on haemodialysis (HD) and peritoneal dialysis (PD), but few studies have addressed this issue. METHODS: Association of modality with incident patients' health status and quality of life scores was investigated with propensity score (PS) analysis and also with traditional multivariable regression analyses. We compared patient reported health status and quality of life scores after 1 year of therapy in 455 HD and 413 PD patients who participated in a national study, stayed on the same modality and had complete socio-demographic and clinical information needed to create a PS indicating their expected probability of starting on PD. RESULTS: One year scores on the majority of health status and quality of life measures were not significantly different for HD and PD patients within propensity-matched quintiles. PD patients' scores were higher than HD patients' scores on effects of kidney disease, burden of kidney disease, staff encouragement and satisfaction with care in some quintiles, and traditional regression analyses confirmed that dialysis modality was associated with patients' scores on these variables. CONCLUSIONS: This study provides support for making the choice of PD more widely available as an option to patients initiating chronic dialysis therapy. Patient lifestyle opportunities associated with use of PD, a home-based and self-care therapy, may also apply to home-based HD or in-centre self-care HD. Patients' expectations regarding treatment and their attitudes toward management of their health may interact with treatment modality to shape patient-reported experience on dialysis; this is an important focus for future studies. PMID- 16046521 TI - Origins of the mutational origin of cancer. PMID- 16046522 TI - CD4+ T-cell death induced by infectious and noninfectious HIV-1: role of type 1 interferon-dependent, TRAIL/DR5-mediated apoptosis. AB - It has been proposed that direct and indirect mechanisms contribute to the unresolved issue of CD4(+) T-cell depletion that results from HIV-1 infection. We recently reported that plasma levels of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) are elevated in HIV-1-infected patients and that they correlate with viral load. The present study investigates the expression of TRAIL death receptor 5 (DR5) in the peripheral-blood mononuclear cells (PBMCs) of HIV-1-infected patients and its role in CD4(+) T-cell death. DR5 expression was elevated and associated with the apoptotic marker annexin V. Apoptosis was reduced in CD4(+) T cells when cultured with anti-DR5 antibody. CD4(+), but not CD8(+), T cells from uninfected donors expressed TRAIL, DR5, and activated caspase-3 when cultured with infectious or noninfectious HIV-1, resulting in preferential apoptosis of CD4(+) T cells. TRAIL, caspase-3 expression, and apoptosis were type 1 interferon (IFN) dependent. Induction of apoptosis and DR5 expression required glycoprotein 120 (gp120)-CD4 interaction. Finally, we analyzed DR5 expression by CD4(+) T cells in highly active antiretroviral therapy (HAART)-treated patients. The decreased viral loads and increased CD4 counts of HAART-responsive patients were associated with a decrease in DR5 mRNA expression by CD4(+) T lymphocytes. We propose a novel model in which a type 1 IFN-regulated TRAIL /DR5 mechanism induces apoptosis of HIV-1-exposed CD4(+) T cells. PMID- 16046523 TI - In vivo analysis of replication and immunogenicity of proviral clones of human T lymphotropic virus type 1 with selective envelope surface-unit mutations. AB - Human T-cell leukemia virus type 1 (HTLV-1) is the causative agent of adult T cell lymphoma/leukemia (ATL). The HTLV-1 envelope gene exhibits limited variability when examined from infected individuals, but has not been tested using infectious clones of the virus in animal models. In vitro assays indicate that HTLV-1 envelope (Env) Ser75Ile, Asn95Asp, and Asn195Asp surface unit (SU) mutants are able to replicate in and immortalize lymphocytes. Herein, we examined the effects of these Env mutants in rabbits inoculated with HTLV-1 immortalized ACH.75, ACH.95, or ACH.195 cell lines (expressing full-length molecular clones with the SU mutations) or the ACH.1 cell line (expressing wild-type SU). All rabbits became infected, and the fidelity of the mutations was maintained throughout the 8-week study. However, SU point mutations resulted in decreased antibody responses to viral group-associated antigen (Gag) and Env antigens. ACH.195 rabbits had a selective decreased antibody response to SU, and one ACH.195 rabbit had an antibody response to both HTLV-1 and HTLV-2 SUs. Some mutant inoculation groups had altered proviral loads. However, peripheral-blood mononuclear cell (PBMC) proviral loads did not correlate with antibody responses. Our data are the first to demonstrate that mutations in critical determinants of HTLV-1 Env SU altered antibody responses and proviral loads, but do not prevent viral replication in vivo. PMID- 16046524 TI - RAS-blocking bisphosphonate zoledronic acid inhibits the abnormal proliferation and differentiation of juvenile myelomonocytic leukemia cells in vitro. AB - Juvenile myelomonocytic leukemia (JMML) is a clonal myeloproliferative/myelodysplastic disorder of early childhood with a poor prognosis. JMML cells are characterized by hypersensitivity to granulocyte macrophage colony-stimulating factor (GM-CSF) caused by a continuously activated GM-CSF receptor-retrovirus-associated sequence (RAS) signal transduction pathway through various molecular mechanisms, resulting in spontaneous GM colony formation in vitro. Bisphosphonate zoledronic acid (ZOL), a RAS-blocking compound, suppressed colony formation from bone marrow (BM) cells of 8 patients with JMML and 5 healthy control subjects without and with GM-CSF (10 ng/mL), respectively, in a dose-dependent manner in clonal culture. At 10 microM ZOL, however, spontaneous GM colony formation from JMML BM cells decreased to 3%, but the formation of G colonies containing granulocytes, but no macrophages, was enhanced, whereas 40% of GM colonies were retained and G colony formation was not affected in culture of normal BM cells with GM-CSF. In suspension culture, cytochemical and flow cytometric analyses showed that 10 microM ZOL also inhibited spontaneous proliferation and differentiation along monocyte/macrophage lineage of JMML BM cells but not the development of normal BM cells by GM-CSF. The inhibitory effect of ZOL on JMML cells was confirmed at a single-clone level and observed even at 3 microM. The current result offers a novel approach to therapy in JMML. PMID- 16046525 TI - CD94 1A transcripts characterize lymphoblastic lymphoma/leukemia of immature natural killer cell origin with distinct clinical features. AB - Most lymphoblastic lymphomas (LBLs) are regarded as neoplasms of immature T cells because they express cytoplasmic CD3 and frequently carry T-cell receptor (TCR) gene rearrangements. Immature natural killer (NK) and T cells, however, have a common bipotent T/NK-cell precursor in the thymus, and NK cells also express cytoplasmic CD3. Thus, some LBLs could arise from immature NK cells. Mature NK cells express 2 CD94 transcripts: 1A, induced by interleukin 15 (IL-15), and 1B constitutively. Because immature NK cells require IL-15 for development, CD94 1A transcripts could be a marker of NK-LBL. To test this hypothesis, we used laser capture microdissection to isolate IL-15 receptor alpha(+) lymphoid cells from the thymus and showed that these cells contained CD94 1A transcripts. We then assessed for CD94 transcripts in 21 cases of LBL that were cytoplasmic CD3(+), nuclear terminal deoxynucleotidyl transferase positive (TdT(+)), and CD56(-), consistent with either the T-cell or NK-cell lineage. We found that 7 LBLs expressed CD94 1A transcripts without TCR gene rearrangements, suggesting NK-cell lineage. Patients with NK-LBL were younger than patients with T-LBL (15 years versus 33 years; P = .11) and had a better 2-year survival (100% versus 27%; P < .01). These results improve the current classification of LBL and contribute to our understanding of NK-cell differentiation. PMID- 16046526 TI - The antileukemia effect of HLA-matched NK and NK-T cells in chronic myelogenous leukemia involves NKG2D-target-cell interactions. AB - To study natural killer (NK) cell-mediated antileukemic activity in chronic myelogenous leukemia (CML), we investigated the ability of HLA-matched and mismatched CD56(+) cells to inhibit granulocyte macrophage-colony-forming unit (CFU-GM) formation by leukemic CD34(+) cells. In 14 HLA-identical donor-recipient pairs, donor CD56(+) cells inhibited CML CFU-GM comparably to effectors from 14 HLA-mismatched unrelated individuals (mean inhibition 42% +/- 9% vs 39.5% +/- 7% at a 10:1 effector-to-target (E/T) ratio), suggesting that killer inhibitory receptor (KIR) incompatibility was not essential for an antileukemic effect. Both CD56(+)CD3(-) (natural killer [NK]) and CD56(+)CD3(+)(NK-T) cells inhibited CFU GM growth of CML but not normal CD34(+) cells. A mechanism for this leukemia specific cytotoxicity was suggested by the abnormal overexpression of major histocompatibility class I chain-related gene A or gene B (MICA/B) on CML CD34 cells and their ability to bind the NK activation ligand NKG2D. However, in vivo, CML cells may avoid NK-cell-mediated immune destruction by immune escape, shedding MICA into the plasma, thereby down-regulating NKG2D on CML CD56(+) cells. PMID- 16046527 TI - Inhibiting the IGF-1 receptor tyrosine kinase with the cyclolignan PPP: an in vitro and in vivo study in the 5T33MM mouse model. AB - Insulin-like growth factor 1 (IGF-1) plays a pleiotropic role in multiple myeloma (MM), that is, in survival, proliferation, chemotaxis, and angiogenesis. Strategies targeting the IGF-1 receptor (IGF-1R) may therefore be important to develop efficient anti-MM agents. In this work we investigated the effect of an IGF-1R tyrosine kinase (IGF-1RTK) inhibitor (picropodophyllin or PPP) in the 5T33MM mouse model. In vitro data showed that PPP reduced IGF-1R autophosphorylation and downstream ERK activation, leading to inhibition of IGF-1 stimulated proliferation and vascular endothelial growth factor (VEGF) secretion of MM cells. In an in vivo study, PPP reduced the bone marrow tumor burden and serum paraprotein in 5T33MM mice by 77% and 90%, respectively, compared to vehicle-treated animals. Angiogenesis was assessed by quantifying the microvessel density on CD31-stained paraffin sections and this was reduced by 60% in the PPP treated group. In a separate survival experiment, Kaplan-Meier analysis demonstrated a significant increase in survival in PPP-treated 5T33MM animals compared to the vehicle controls (28 versus 18 days). These data suggest that the IGF-1RTK inhibitor PPP possesses a marked antitumor activity and strongly points to the possibility of using IGF-1R inhibitors in the treatment of MM. PMID- 16046528 TI - Prevalence, clinical profile, and prognosis of NPM mutations in AML with normal karyotype. AB - Mutation of the nucleophosmin (NPM) gene has been reported as the most frequent mutation in acute myeloid leukemia (AML), especially in the presence of a normal karyotype. In this subgroup of intermediate-risk AML, the identification of other gene mutations (eg, FLT3, CCAAT/enhancer-binding protein-alpha [CEBPA]) has helped to refine the prognosis. This study explored the prevalence and the prognostic impact of NPM mutations in a cohort of 106 patients with normal karyotype AML. NPM exon 12 mutations were detected by polymerase chain reaction (PCR) and fragment analysis for the insertion/deletion globally resulting in a 4 bp insertion. NPM mutations were detected in 47% of patients and were associated with a high white blood cell count, involvement of the monocytic lineage (M4/M5), and a decreased prevalence of CEBPA mutations. Complete remission rate and long term outcome did not differ between NPM-mutated and -nonmutated patients. Prospective studies are needed to confirm the definitive place of NPM mutation detection to predict AML response to therapy. PMID- 16046529 TI - Cytomegalovirus MCK-2 controls mobilization and recruitment of myeloid progenitor cells to facilitate dissemination. AB - Murine cytomegalovirus encodes a secreted, pro-inflammatory chemokine-like protein, MCK-2, that recruits leukocytes and facilitates viral dissemination. We have shown that MCK-2-enhanced recruitment of myelomonocytic leukocytes with an immature phenotype occurs early during infection and is associated with efficient viral dissemination. Expression of MCK-2 drives the mobilization of a population of leukocytes from bone marrow that express myeloid marker Mac-1 (CD11b), intermediate levels of Gr-1 (Ly6 G/C), platelet-endothelial-cell adhesion molecule-1 (PECAM-1, CD31), together with heterogeneous levels of stem-cell antigen-1 (Sca-1, Ly-6 A /E). Recombinant MCK-2 mediates recruitment of this population even in the absence of viral infection. Recruitment of this cell population and viral dissemination via the bloodstream to salivary glands proceeds normally in mice that lack CCR2 and MCP-1 (CCL2), suggesting that recruitment of macrophages is not a requisite component of pathogenesis. Thus, a systemic impact of MCK-2 enhances the normal host response and causes a marked increase in myelomonocytic recruitment with an immature phenotype to initial sites of infection. Mobilization influences levels of virus dissemination via the bloodstream to salivary glands and is dependent on a myelomonocytic cell type other than mature macrophages. PMID- 16046530 TI - Influence of T-cell depletion on chronic graft-versus-host disease: results of a multicenter randomized trial in unrelated marrow donor transplantation. AB - Donor-derived T cells have been proposed to play a role in pathogenesis of chronic graft-versus-host disease (cGVHD). The impact of ex vivo T-cell depletion (TCD) on cGVHD was analyzed in a randomized multicenter trial involving unrelated donor marrow transplants. A total of 404 patients diagnosed with hematologic malignancies received a total body irradiation-based myeloablative conditioning regimen. GVHD prophylaxis included TCD plus cyclosporine (CSA) or unmodified grafts with CSA plus methotrexate (M/C). Median recipient age was 31.2 years (range, 0.5-55.6 years); median follow-up time since randomization was 4.2 years. The mean number of T cells infused was 1 log lower on the TCD arm. The incidence of cGVHD at 2 years was similar between the TCD and M/C arms, 29% versus 34% (P = .27), respectively. Survival at 3 years from diagnosis of cGVHD was also similar, (TCD 51% versus M/C 58%; P = .29). The proportion of patients with cGVHD who discontinued immunosuppression at 5 years was not different (TCD 72% versus M/C 63%; P = .27), and incidence of serious infections and leukemia relapse were similar on both treatment arms. In spite of a significant reduction of acute GVHD, TCD did not reduce the incidence of cGVHD or improve survival in patients who developed cGVHD. PMID- 16046531 TI - Plasmodium falciparum rhoptry protein RSP2 triggers destruction of the erythroid lineage. AB - The destruction of erythrocytes and defects in erythropoiesis are among the most frequently observed causes of morbidity in severe Plasmodium falciparum malaria. The molecular mechanisms involved remain unclear, despite extensive investigation. We show here, for the first time, that tagging with the parasite rhoptry protein ring surface protein 2 (RSP2) is not restricted to the surfaces of normal erythrocytes, as previously reported, but that it extends to erythroid precursor cells in the bone marrow of anemic malaria patients. Monoclonal mouse antibodies and human sera from patients with severe anemia, reacting with RSP2 tagged erythrocytes, induced cell destruction by phagocytosis and complement activation in vitro. Our observations reveal a new parasite mechanism implicated in the destruction of normal erythrocytes and probably dyserythropoiesis in malaria patients. These data suggest that the tagging of host cells with RSP2 may trigger anemia in falciparum malaria. PMID- 16046532 TI - Diffuse large B-cell lymphoma subgroups have distinct genetic profiles that influence tumor biology and improve gene-expression-based survival prediction. AB - Gene-expression profiling has identified 3 major subgroups of diffuse large B cell lymphoma (DLBCL): germinal center B-cell-like (GCB), activated B-cell-like (ABC), and primary mediastinal DLBCL (PMBCL). Using comparative genomic hybridization (CGH), we investigated the genetic alterations of 224 cases of untreated DLBCL (87 GCB-DLBCL, 77 ABC-DLBCL, 19 PMBCL, and 41 unclassified DLBCL) previously characterized by gene-expression profiling. The DLBCL subgroups differed significantly in the frequency of particular chromosomal aberrations. ABC-DLBCL had frequent trisomy 3, gains of 3q and 18q21-q22, and losses of 6q21 q22, whereas GCB-DLBCL had frequent gains of 12q12, and PMBCL had gains of 9p21 pter and 2p14-p16. Parallel analysis of CGH alterations, locus-specific gene expression profiles, and global gene-expression signatures revealed that DNA amplifications and gains had a substantial impact on the expression of genes in the involved chromosomal regions, and some genes were overexpressed in a DLBCL subgroup-specific fashion. Unexpectedly, specific chromosomal alterations were associated with significant changes in gene-expression signatures that reflect various aspects of lymphoma cell biology as well as the host response to the lymphoma. In addition, gains involving the chromosomal region 3p11-p12 provided prognostic information that was statistically independent of the previously defined gene-expression-based survival model, thereby improving its predictive power. PMID- 16046533 TI - Targeting MLL-AF4 with short interfering RNAs inhibits clonogenicity and engraftment of t(4;11)-positive human leukemic cells. AB - The chromosomal translocation t(4;11) marks infant acute lymphoblastic leukemia associated with a particularly dismal prognosis. The leukemogenic role of the corresponding fusion gene MLL-AF4 is not well understood. We show that transient inhibition of MLL-AF4 expression with small interfering RNAs impairs the proliferation and clonogenicity of the t(4; 11)-positive human leukemic cell lines SEM and RS4;11. Reduction of mixed-lineage leukemia (MLL)-ALL-1 fused gene from chromosome 4 (AF4) levels induces apoptosis associated with caspase-3 activation and diminished BCL-X(L) expression. Suppression of MLL-AF4 is paralleled by a decreased expression of the homeotic genes HOXA7, HOXA9, and MEIS1. MLL-AF4 depletion inhibits expression of the stem-cell marker CD133, indicating hematopoietic differentiation. Transfection of leukemic cells with MLL AF4 siRNAs reduces leukemia-associated morbidity and mortality in SCID mice that received a xenotransplant, suggesting that MLL-AF4 depletion negatively affects leukemia-initiating cells. Our findings demonstrate that MLL-AF4 is important for leukemic clonogenicity and engraftment of this highly aggressive leukemia. Targeted inhibition of MLL-AF4 fusion gene expression may lead to an effective and highly specific treatment of this therapy-resistant leukemia. PMID- 16046534 TI - Tropical mountain cradles of dry forest diversity. PMID- 16046535 TI - Functional conformational motions in the turnover cycle of cholesterol oxidase. AB - Reexamining experimental data of single-molecule fluorescence correlation spectroscopy for cholesterol oxidase, we find that the existing Michaelis-Menten models with dynamical disorder cannot explain strong correlations between subsequent turnover cycles revealed in the diagonal feature in the joint statistical distribution of adjacent "on" times of this enzyme. We suggest that functional conformational motions representing ordered sequences of transitions between a set of conformational substates are involved, along with equilibrium conformational fluctuations in the turnover cycle of cholesterol oxidase. A two channel model of single-enzyme dynamics, including a slow functional conformational motion in one of the channels, is proposed that allows us to reproduce such strong correlations. PMID- 16046536 TI - Lipotechoic acid in lactobacilli: D-alanine makes the difference. PMID- 16046537 TI - Obesity and metabolic perturbations after loss of aquaporin 7, the adipose glycerol transporter. PMID- 16046538 TI - Inhibition of drug-resistant mutants of ABL, KIT, and EGF receptor kinases. AB - To realize the full potential of targeted protein kinase inhibitors for the treatment of cancer, it is important to address the emergence of drug resistance in treated patients. Mutant forms of BCR-ABL, KIT, and the EGF receptor (EGFR) have been found that confer resistance to the drugs imatinib, gefitinib, and erlotinib. The mutations weaken or prevent drug binding, and interestingly, one of the most common sites of mutation in all three kinases is a highly conserved "gatekeeper" threonine residue near the kinase active site. We have identified existing clinical compounds that bind and inhibit drug-resistant mutant variants of ABL, KIT, and EGFR. We found that the Aurora kinase inhibitor VX-680 and the p38 inhibitor BIRB-796 inhibit the imatinib- and BMS-354825-resistant ABL(T315I) kinase. The KIT/FLT3 inhibitor SU-11248 potently inhibits the imatinib-resistant KIT(V559D/T670I) kinase, consistent with the clinical efficacy of SU-11248 against imatinib-resistant gastrointestinal tumors, and the EGFR inhibitors EKB 569 and CI-1033, but not GW-572016 and ZD-6474, potently inhibit the gefitinib- and erlotinib-resistant EGFR(L858R/T790M) kinase. EKB-569 and CI-1033 are already in clinical trials, and our results suggest that they should be considered for testing in the treatment of gefitinib/erlotinib-resistant non-small cell lung cancer. The results highlight the strategy of screening existing clinical compounds against newly identified drug-resistant mutant variants to find compounds that may serve as starting points for the development of next generation drugs, or that could be used directly to treat patients that have acquired resistance to first-generation targeted therapy. PMID- 16046539 TI - A different point of hue. PMID- 16046540 TI - NK cytotoxicity against CD4+ T cells during HIV-1 infection: a gp41 peptide induces the expression of an NKp44 ligand. AB - HIV infection leads to a state of chronic immune activation and progressive deterioration in immune function, manifested most recognizably by the progressive depletion of CD4+ T cells. A substantial percentage of natural killer (NK) cells from patients with HIV infection are activated and express the natural cytotoxicity receptor (NCR) NKp44. Here we show that a cellular ligand for NKp44 (NKp44L) is expressed during HIV-1 infection and is correlated with both the progression of CD4+ T cell depletion and the increase of viral load. CD4+ T cells expressing this ligand are highly sensitive to the NK lysis activity mediated by NKp44+ NK cells. The expression of NKp44L is induced by the linear motif NH2 SWSNKS-COOH of the HIV-1 envelope gp41 protein. This highly conserved motif appears critical to the sharp increase in NK lysis of CD4+ T cells from HIV infected patients. These studies strongly suggest that induction of NKp44L plays a key role in the lysis of CD4+ T cells by activated NK cells in HIV infection and consequently provide a framework for considering how HIV-1 may use NK cell immune surveillance to trigger CD4+ T cells. Understanding this mechanism may help to develop future therapeutic strategies and vaccines against HIV-1 infection. PMID- 16046541 TI - Trehalose 6-phosphate regulates starch synthesis via posttranslational redox activation of ADP-glucose pyrophosphorylase. AB - Trehalose is the most widespread disaccharide in nature, occurring in bacteria, fungi, insects, and plants. Its precursor, trehalose 6-phosphate (T6P), is also indispensable for the regulation of sugar utilization and growth, but the sites of action are largely unresolved. Here we use genetic and biochemical approaches to investigate whether T6P acts to regulate starch synthesis in plastids of higher plants. Feeding of trehalose to Arabidopsis leaves led to stimulation of starch synthesis within 30 min, accompanied by activation of ADP-glucose pyrophosphorylase (AGPase) via posttranslational redox modification. The response resembled sucrose but not glucose feeding and depended on the expression of SNF1 related kinase. We also analyzed transgenic Arabidopsis plants with T6P levels increased by expression of T6P synthase or decreased by expression of T6P phosphatase (TPP) in the cytosol. Compared with wild type, leaves of T6P synthase expressing plants had increased redox activation of AGPase and increased starch, whereas TPP-expressing plants showed the opposite. Moreover, TPP expression prevented the increase in AGPase activation in response to sucrose or trehalose feeding. Incubation of intact isolated chloroplasts with 100 muM T6P significantly and specifically increased reductive activation of AGPase within 15 min. Results provide evidence that T6P is synthesized in the cytosol and acts on plastidial metabolism by promoting thioredoxin-mediated redox transfer to AGPase in response to cytosolic sugar levels, thereby allowing starch synthesis to be regulated independently of light. The discovery informs about the evolution of plant metabolism and how chloroplasts of prokaryotic origin use an intermediate of the ancient trehalose pathway to report the metabolic status of the cytosol. PMID- 16046542 TI - Engineering a signal transduction mechanism for protein-based biosensors. AB - Hybridization-induced conformational changes have been successfully used in biosensors for the transduction of DNA-binding events into readily observable optical or electronic signals. Similar signal transduction has not, however, proven of equal utility in protein-based biosensors. The discrepancy arises because, unlike ssDNA, most proteins do not undergo significant conformational changes upon ligand binding. Here, we describe a solution to this problem. We show that an arbitrarily selected, normally well folded protein can be rationally engineered such that it undergoes ligand-induced folding. The engineered protein responds rapidly (milliseconds) and selectively to its target, and it couples recognition with the largest possible conformational change: folding. These traits suggest that ligand-induced folding could serve as an ideal signal transduction mechanism. Consistent with this claim, we demonstrate a label-free optical biosensor based on the effect that is sufficiently selective to detect its target even in complex, contaminant-ridden samples such as blood serum. PMID- 16046544 TI - Molecular structure as a blueprint for supramolecular structure chemistry in confined spaces. PMID- 16046543 TI - Crystalline molecular machines: encoding supramolecular dynamics into molecular structure. AB - Crystalline molecular machines represent an exciting new branch of crystal engineering and materials science with important implications to nanotechnology. Crystalline molecular machines are crystals built with molecules that are structurally programmed to respond collectively to mechanic, electric, magnetic, or photonic stimuli to fulfill specific functions. One of the main challenges in their construction derives from the picometric precision required for their mechanic operation within the close-packed, self-assembled environment of crystalline solids. In this article, we outline some of the general guidelines for their design and apply them for the construction of molecular crystals with units intended to emulate macroscopic gyroscopes and compasses. Recent advances in the preparation, crystallization, and dynamic characterization of these interesting systems offer a foothold to the possibilities and help highlight some avenues for future experimentation. PMID- 16046545 TI - Jade-1, a candidate renal tumor suppressor that promotes apoptosis. AB - Medical therapies are lacking for advanced renal cancer, so there is a great need to understand its pathogenesis. Most renal cancers have defects in the von Hippel Lindau tumor suppressor pVHL. The mechanism by which pVHL protein functions in renal tumor suppression remains unclear. Jade-1 is a short-lived, kidney-enriched transcription factor that is stabilized by direct interaction with pVHL. Loss of Jade-1 stabilization by pVHL correlates with renal cancer risk, making the relationship between Jade-1 and renal cancer compelling. We report that Jade-1 expression was barely detectable in all tested renal cancer cell lines, regardless of VHL status. Strikingly, proteasome inhibitor treatment increased endogenous Jade-1 expression up to 10-fold. Jade-1 inhibited renal cancer cell growth, colony formation, and tumor formation in nude mice. Intriguingly, Jade-1 also affected the pattern of cell growth in monolayer culture and 3D culture. Jade-1 increased apoptosis by 40-50% and decreased levels of antiapoptotic Bcl-2. Antisense Jade-1-expressing cells confirmed these results. Therefore, Jade-1 may suppress renal cancer cell growth in part by increasing apoptosis. Jade-1 may represent a proapoptotic barrier to proliferation that must be overcome generally in renal cancer, perhaps initially by pVHL inactivation and subsequently by increased proteasomal activity. Therefore, Jade-1 may be a renal tumor suppressor. PMID- 16046546 TI - Epidemiological evidence of an early wave of the 1918 influenza pandemic in New York City. AB - The 1918 "Spanish flu" was the fastest spreading and most deadly influenza pandemic in recorded history. Hypotheses of its origin have been based on a limited collection of case and outbreak reports from before its recognized European emergence in the summer of 1918. These anecdotal accounts, however, remain insufficient for determining the early diffusion and impact of the pandemic virus. Using routinely collected monthly age-stratified mortality data, we show that an unmistakable shift in the age distribution of epidemic deaths occurred during the 1917/1918 influenza season in New York City. The timing, magnitude, and age distribution of this mortality shift provide strong evidence that an early wave of the pandemic virus was present in New York City during February-April 1918. PMID- 16046547 TI - Transfection of keratin 18 gene in human breast cancer cells causes induction of adhesion proteins and dramatic regression of malignancy in vitro and in vivo. AB - This study shows that high keratin 18 (K18) expression in tumor cells is associated with reduced invasiveness in vitro and lack of tumorigenicity in nude mice. We previously showed that high K18 expression correlated with a good prognosis and that reducing K18 expression increased the aggressiveness of established breast cancer cell lines. To confirm these observations, we transfected the human K18 gene into the human breast cancer cell line MDA-MB-231 and isolated a stable overexpressing clone. The forced K18 expression was associated with a complete loss of the previously strong vimentin expression in the parent cell line, induction of the K18 dimerization partner K8, and up regulation of adhesion proteins. These changes were accompanied by a dramatic reduction in the aggressiveness of the K18 transfectants in vitro and in vivo. We conclude that forced reexpression of K18 causes at least partial redifferentiation of the tumor cell, followed by a corresponding regression of malignant phenotype. PMID- 16046548 TI - Lipopolysaccharide prevents doxorubicin-induced apoptosis in RAW 264.7 macrophage cells by inhibiting p53 activation. AB - The effect of lipopolysaccharide on doxorubicin-induced cell death was studied by using mouse RAW 264.7 macrophage cells. Pretreatment with lipopolysaccharide at 10 ng/mL prevented doxorubicin-induced cell death and the inhibition was roughly dependent on the concentration of lipopolysaccharide. Posttreatment with lipopolysaccharide for 1 hour also prevented doxorubicin-induced cell death. Lipopolysaccharide inhibited DNA fragmentation and caspase-3 activation in doxorubicin-treated RAW 264.7 cells, suggesting the prevention of doxorubicin induced apoptosis. Lipopolysaccharide did not significantly inhibit doxorubicin induced DNA damage detected by single-cell gel electrophoresis (comet) assay. Lipopolysaccharide definitely inhibited the stabilization and nuclear translocation of p53 in doxorubicin-treated RAW 264.7 cells. Lipopolysaccharide, as well as being an inhibitor of p53, abolished doxorubicin-induced apoptosis. Therefore, p53 was suggested to play a pivotal role in the prevention of doxorubicin-induced apoptosis in RAW 264.7 cells by lipopolysaccharide. PMID- 16046549 TI - Proteasome structures affected by ionizing radiation. AB - Exposure of cells to ionizing radiation slows the rate of degradation of substrates through the proteasome. Because the 26S proteasome degrades most short lived cellular proteins, changes in its activity might significantly, and selectively, alter the life span of many signaling proteins and play a role in promoting the biological consequences of radiation exposure, such as cell cycle arrest, DNA repair, and apoptosis. Experiments were therefore undertaken to identify the radiation target that is associated with the proteasome. Regardless of whether they were irradiated before or after extraction and purification from human prostate cancer PC3 cells, 26S proteasomes remained intact but showed a rapid 30% to 50% dose-independent decrease in their three major enzymatic activities following exposure to 1 to 20 Gy. There was no effect on 20S proteasomes, suggesting that the radiation-sensitive target is located in the 19S cap of the 26S proteasome, rather than in the enzymatically active core. Because the base of the 19S cap contains an ATPase ring that mediates substrate unfolding, pore opening, and translocation of substrates into the catalytic chamber, we examined whether the ATPase activity of purified 26S proteasomes was affected. In fact, in vitro irradiation of proteasomes enhanced their ATPase activity. Furthermore, pretreatment with low concentrations of the free radical scavenger tempol was able to prevent both the radiation-induced decrease in proteolytic activity and the increase in ATP utilization, indicating that free radicals are mediators of these radiation-induced phenomena. Finally, we have shown that cell irradiation results in the accumulation of proteasome substrates: polyubiquitinated proteins and ornithine decarboxylase, indicating that the observed decrease in proteasome function is physiologically relevant. PMID- 16046550 TI - Phosphorylation of AML1/RUNX1 regulates its degradation and nuclear matrix association. AB - The acute myeloid leukemia 1 (AML1) transcription factors are key regulators of hematopoietic differentiation. Cellular AML1c protein is found in the nucleus and can be separated into two fractions, one soluble in buffers containing salt and nonionic detergent and the other insoluble and tightly bound to the nuclear matrix. We find that the AML1c protein is modified by both phosphorylation and ubiquitination. Our studies show that the majority of the ubiquitinated AML1c is associated with the insoluble nuclear matrix. Treatment of cells with the proteasome inhibitor PS341 (Velcade, Bortezomib) increases the levels of ubiquitinated AML1c. Mutation of the four phosphorylation sites necessary for transcriptional regulation (serine 276, serine 293, serine 303, and threonine 300) mimics the effects of the proteasome inhibitor, increasing the levels of ubiquitinated, matrix-bound AML1c. We find that the soluble and insoluble forms of AML1c are degraded at a similar rate. However, mutation of these four serine/threonine residues statistically increases the half-life of the matrix associated AML1c. Thus, phosphorylation of AML1c on specific serine/threonine residues controls both transcriptional activity and rate of degradation. PMID- 16046551 TI - Transforming growth factor-alpha inhibits the intrinsic pathway of c-Myc-induced apoptosis through activation of nuclear factor-kappaB in murine hepatocellular carcinomas. AB - Nuclear factor-kappaB (NF-kappaB) plays an important role during liver neoplastic development through transcriptional regulation of prosurvival genes, which then counteract the death-inducing signals elicited by the host immune response. The c Myc proto-oncogene is frequently deregulated in liver tumors. Furthermore, enforced expression of c-Myc in the liver promotes the development of hepatocellular carcinomas, a process that is accelerated by coexpression with transforming growth factor-alpha (TGF-alpha). TGF-alpha/c-Myc-derived hepatocellular carcinomas display reduced apoptotic levels compared with those of single c-Myc transgenic hepatocellular carcinomas, suggesting that TGF-alpha provides a survival advantage to c-Myc-transformed hepatocytes. Given that TGF alpha/c-Myc hepatocellular carcinomas display constitutive NF-kappaB activity, here, we have tested the hypothesis that enforced expression of TGF-alpha results in constitutive NF-kappaB activation and enhanced cell survival using TGF-alpha/c Myc-derived hepatocellular carcinoma cell lines. We show that TGF-alpha induces NF-kappaB through the phosphatidylinositol 3-kinase/Akt axis in these bitransgenic hepatocellular carcinomas. Furthermore, we found that adenovirus mediated inhibition of NF-kappaB activity impairs the ability of TGF-alpha/c-Myc derived tumor cells to grow in an anchorage-independent fashion due to sensitization to c-Myc-induced apoptosis. Lastly, we show that NF-kappaB inhibits c-Myc-induced activation of caspase-9 and caspase-3 through up-regulation of the antiapoptotic target genes Bcl-X(L) and X-linked inhibitor of apoptosis (XIAP). Overall, these results underscore a crucial role of NF-kappaB in disabling apoptotic pathways initiated by oncogenic transformation. PMID- 16046552 TI - Molecular consequences of silencing mutant K-ras in pancreatic cancer cells: justification for K-ras-directed therapy. AB - Mutation of the K-ras gene is an early event in the development of pancreatic adenocarcinoma and, therefore, RNA interference (RNAi) directed toward mutant K ras could represent a novel therapy. In this study, we examine the phenotypic and molecular consequences of exposure of pancreatic tumor cells to mutant-specific K ras small interfering RNA. Specific reduction of activated K-ras via RNAi in Panc 1 and MiaPaca-2 cells resulted in cellular changes consistent with a reduced capacity to form malignant tumors. These changes occur through distinct mechanisms but likely reflect an addiction of each cell line to oncogene stimulation. Both cell lines show reduced proliferation after K-ras RNAi, but only MiaPaca-2 cells showed increased apoptosis. Both cell lines showed reduced migration after K-ras knockdown, but changes in integrin levels were not consistent between the cell lines. Both cell lines showed alteration of the level of GLUT-1, a metabolism-associated gene that is downstream of c-myc, with Panc-1 cells demonstrating decreased GLUT-1 levels, whereas MiaPaca-2 cells showed increased levels of expression after K-ras knockdown. Furthermore, after K-ras RNAi, there was a reduction in angiogenic potential of both Panc-1 and MiaPaca-2 cells. Panc-1 cells increased the level of expression of thrombospondin-1, an endogenous inhibitor of angiogenesis, whereas MiaPaca-2 cells decreased the production of vascular endothelial growth factor, a primary stimulant of angiogenesis in pancreatic tumors. We have found that silencing mutant K-ras through RNAi results in alteration of tumor cell behavior in vitro and suggests that targeting mutant K-ras specifically might be effective against pancreatic cancer in vivo. PMID- 16046553 TI - Up-regulation of prostaglandin biosynthesis by leukotriene C4 in elicited mice peritoneal macrophages activated with lipopolysaccharide/interferon-{gamma}. AB - Leukotrienes (LT) and prostaglandins (PG) are proinflammatory mediators generated by the conversion of arachidonic acid via 5-lipoxygenase (5-LO) and cyclooxygenase (COX) pathways. It has long been proposed that the inhibition of the 5-LO could enhance the COX pathway leading to an increased PG generation. We have found that in in vitro models of inflammation, such as mice-elicited peritoneal macrophages activated with lipopolysaccharide (LPS)/interferon-gamma (IFN-gamma), the deletion of the gene encoding for 5-LO or the enzyme activity inhibition corresponded to a negative modulation of the COX pathway. Moreover, exogenously added LTC(4), but not LTD(4), LTE(4), and LTB(4), was able to increase PG production in stimulated cells from 5-LO wild-type and knockout mice. LTC(4) was not able to induce COX-2 expression by itself but rather potentiated the action of LPS/IFN-gamma through the extracellular signal-regulated kinase-1/2 activation, as demonstrated by the use of a specific mitogen-activated protein kinase (MAPK) kinase inhibitor. The LT-induced increase in PG generation, as well as MAPK activation, was dependent by a specific ligand-receptor interaction, as demonstrated by the use of a cys-LT1 receptor antagonist, although also a direct action of the antagonist used, on PG generation, cannot be excluded. Thus, the balance between COX and 5-LO metabolites could be of great importance in controlling macrophage functions and consequently, inflammation and tumor promotion. PMID- 16046554 TI - Differential expression of function-related antigens on blood monocytes in children with hemolytic uremic syndrome. AB - Monocytes (Mo) mediate central functions in inflammation and immunity. Different subpopulations of Mo with distinct phenotype and functional properties have been described. Here, we investigate the phenotype and function of peripheral Mo from children with hemolytic uremic syndrome (HUS). For this purpose, blood samples from patients in the acute period of HUS (HUS AP) were obtained on admission before dialysis and/or transfusion. The Mo phenotypic characterization was performed on whole blood by flow cytometry, and markers associated to biological functions were selected: CD14 accounting for lipopolysaccharide (LPS) responsiveness, CD11b for adhesion, Fc receptor for immunoglobulin G type I (FcgammaRI)/CD64 for phagocytosis and cytotoxicity, and human leukocyte antigen (HLA)-DR for antigen presentation. Some of these functions were also determined. Moreover, the percentage of CD14+ CD16+ Mo was evaluated. We found that the entire HUS AP Mo population exhibited reduced CD14, CD64, and CD11b expression and decreased LPS-induced tumor necrosis factor production and Fcgamma-dependent cytotoxicity. HUS AP showed an increased percentage of CD14+ CD16+ Mo with higher CD16 and lower CD14 levels compared with the same subset from healthy children. Moreover, the CD14++ CD16- Mo subpopulation of HUS AP had a decreased HLA-DR expression, which correlated with severity. In conclusion, the Mo population from HUS AP patients presents phenotypic and functional alterations. The contribution to the pathogenesis and the possible scenarios that led to these changes are discussed. PMID- 16046555 TI - Energy response of an aluminium oxide detector in kilovoltage and megavoltage photon beams: an EGSnrc Monte Carlo simulation study. AB - A Monte Carlo study of the energy response of an aluminium oxide (Al(2)O(3)) detector in kilovoltage and megavoltage photon beams relative to (60)Co gamma rays has been performed using EGSnrc Monte Carlo simulations. The sensitive volume of the Al(2)O(3) detector was simulated as a disc of diameter 2.85 mm and thickness 1 mm. The phantom material was water and the irradiation depth chosen was 2.0 cm in kilovoltage photon beams and 5.0 cm in megavoltage photon beams. The results show that the energy response of the Al(2)O(3) detector is constant within 3% for photon beam energies in the energy range of (60)Co gamma rays to 25 MV X rays. However, the Al(2)O(3) detector shows an enhanced energy response for kilovoltage photon beams, which in the case of 50 kV X rays is 3.2 times higher than that for (60)Co gamma rays. There is essentially no difference in the energy responses of LiF and Al(2)O(3) detectors irradiated in megavoltage photon beams when these Al(2)O(3) results are compared with literature data for LiF thermoluminescence detectors. However, the Al(2)O(3) detector has a much higher enhanced response compared with LiF detectors in kilovoltage X-ray beams, more than twice as much for the case of 50 kV X rays. PMID- 16046556 TI - Rat testis as a radiobiological in vivo model for radionuclides. AB - The radiobiological effect of intracellularly localised radionuclides emitting low energy electrons (Auger electrons) has received much attention. Most in vivo studies reported have been performed in the mouse testis. We have investigated the rat testis as an in vivo radiobiological model, with sperm-head survival, testis weight loss and also alteration in the blood plasma hormone levels of FSH and LH as radiobiological endpoints. Validation of the rat testis model was evaluated by using mean absorbed doses of up to 10 Gy from intratesticularly (i.t.) injected (111)In oxine or local X-ray irradiation. Biokinetics of the i.t. injected radionuclide was analysed by scintillation camera imaging and used in the absorbed dose estimation. By the analysis of the autoradiographs, the activity distribution was revealed. Cell fractionation showed (111)In to be mainly associated with the cell nuclei. External irradiations were monitored by thermoluminescence dosimeters. The sperm-head survival was the most sensitive radiobiological parameter correlated to the mean absorbed dose, with a D(37) of 2.3 Gy for (111)In oxine and 1.3 Gy for X rays. The levels of plasma pituitary gonadal hormones FSH and LH were elevated for absorbed doses >7.7 Gy. This investigation shows that the radiobiological model based on the rat testis has several advantages compared with the previously commonly used mouse testis model. The model is appropriate for further investigations of basic phenomena such as radiation geometry, intracellular kinetics and heterogeneity, crucial for an understanding of the biological effect of low-energy electrons. PMID- 16046557 TI - Determination of radon concentrations of the Dikili geothermal area in western Turkey. AB - For this study, a geothermal area around Izmir-Dikili, located in the western part of Turkey, was chosen as a measuring site and the radon concentrations of environmental samples were determined. Indoor radon monitoring was performed for 3 months in dwellings located in different part of the region using passive CR-39 nuclear track detectors. The radon concentrations of water samples drawn from wells, municipal supplies, village fountains and spas in the area were measured using a liquid scintillation detector (Packard Tri-Carb 2770 TR/SL, A-277001). Soil samples collected from each location were analysed and (226)Ra concentrations were determined using a gamma ray spectrometer connected to an HPGe detector. Annual effective doses from radon inhalation and ingestion were calculated. PMID- 16046558 TI - Comparison doses of secondary neutron with the heavy ions in a 75-Mev/n heavy ion beam. AB - The angular distributions for neutrons of energy >6 MeV that are induced by 75 MeV/n 12C6+ and 16O8+ ions were measured with the activation method of Al threshold detectors at the radiobiological terminal of HIRFL. The data were obtained by a high-purity Ge(HpGe) detector. The results show that the neutron angular distributions produced by heavy ion beams are strongly peaked in the forward direction and decreased exponentially with angles in experimental area. The experimental conditions for these measurements were similar to those for biological experiments, so the results should be representative of neutrons produced by heavy ions during the biological experiments and tumour therapy. Comparing with the neutron doses produced by the heavy ion beam, the heavy ion dose is the main factor in biological effects and tumour therapy response, so the contribution of neutron dose can be neglected. PMID- 16046559 TI - Prehypertension, patient outcomes, and the knowledge base of family medicine. PMID- 16046560 TI - Prehypertension and cardiovascular morbidity. AB - PURPOSE: The Seventh Report of the Joint National Commission (JNC 7) on High Blood Pressure established prehypertension (120 to 139 mm Hg systolic or 80 to 89 mm Hg diastolic) as a new risk category. We aim to determine the risk of major cardiovascular events associated with blood pressure in the prehypertensive range in a longitudinal, population-based cohort. METHODS: Analyses were conducted on participants in the National Health and Nutrition Examination Survey I (1971 1975) observed for 18 years for major cardiovascular disease events. Cox proportional hazard ratios were calculated to assess relative risk of cardiovascular disease, including stroke, myocardial infarction, and heart failure, in participants with prehypertension and normal blood pressure (<120/80 mm Hg). RESULTS: Prehypertension was associated with increased risk for cardiovascular disease (1.79 [95% confidence interval (CI) 1.40-2.24]) in unadjusted analysis. After adjustment for cardiovascular risk factors, the relationship of prehypertension to cardiovascular disease was diminished but persisted (1.32 [95% CI 1.05-1.65]). Ninety-three percent of prehypertensive individuals had at least 1 cardiovascular risk factor. Low prehypertension (120 129/80-84 mm Hg) was associated with increased cardiovascular disease in unadjusted analyses (1.56 [95% CI 1.23-1.98]) but was not statistically significant in adjusted analyses (1.24 [95% CI 0.96-1.59]). High-normal blood pressure (130-139/85-89 mm Hg) remained a predictor of cardiovascular disease in unadjusted (2.13 [95% CI 1.64-2.76]) and adjusted (1.42 [95% CI 1.09-1.84]) analyses. CONCLUSIONS: In a longitudinal, population-based, US cohort, prehypertension was associated with increased risk of major cardiovascular events independently of other cardiovascular risk factors. These findings, along with the presence of cardiovascular risk factors in the majority of participant sample with prehypertension, support recommendations for physicians to actively target lifestyle modifications and multiple risk reduction in their prehypertensive patients. PMID- 16046561 TI - Impact of an electronic medical record on diabetes quality of care. AB - PURPOSE: This study was designed to evaluate the impact of electronic medical record (EMR) implementation on quality of diabetes care. METHODS: We conducted a 5-year longitudinal study of 122 adults with diabetes mellitus at an intervention (EMR) clinic and a comparison (non-EMR) clinic. Clinics had similarly trained primary care physicians, similar patient populations, and used a common diabetes care guideline that emphasized the importance of glucose control. The EMR provided basic decision support, including prompts and reminders for diabetes care. Preintervention and postintervention frequency of testing for glycated hemoglobin (HbA1c) and low-density lipoprotein (LDL) levels were compared with and without adjustment for patient age, sex, comorbidity, and baseline HbA1c level. RESULTS: Frequency of HbA1c tests increased at the EMR clinic compared with the frequency at the non-EMR clinic (P < .001). HbA1c levels improved in both clinics (P < .05) with no significant differences between clinics 2 years (P = .10) or 4 years (P = .27) after EMR implementation. Similar results were observed for LDL levels. CONCLUSIONS: In this controlled study, EMR use led to an increased number of HbA1c and LDL tests but not to better metabolic control. If EMRs are to fulfill their promise as care improvement tools, improved implementation strategies and more sophisticated clinical decision support may be needed. PMID- 16046562 TI - Implementing an electronic medical record in a family medicine practice: communication, decision making, and conflict. AB - PURPOSE: Electronic medical record (EMR) systems offer substantial opportunities to organize and manage clinical data in ways that can potentially improve preventive health care, the management of chronic illness, and the financial health of primary care practices. The functionality of EMRs as implemented, however, can vary substantially from that envisaged by their designers and even from those who purchase the programs. The purpose of this study was to explore how unique aspects of a family medicine office culture affect the initial implementation of an EMR. METHODS: As part of a larger study, we conducted a qualitative case study of a private family medicine practice that had recently purchased and implemented an EMR. We collected data using participant observation, in-depth interviews, and key informant interviews. After the initial data collection, we shared our observations with practice members and returned 1 year later to collect additional data. RESULTS: Dysfunctional communication patterns, the distribution of formal and informal decision-making power, and internal conflicts limited the effective implementation and use of the EMR. The implementation and use of the EMR made tracking and monitoring of preventive health and chronic illness unwieldy and offered little or no improvement when compared with paper charts. CONCLUSIONS: Implementing an EMR without an understanding of the systemic effects and communication and the decision-making processes within an office practice and without methods for bringing to the surface and addressing conflicts limits the opportunities for improved care offered by EMRs. Understanding how these common issues manifest within unique practice settings can enhance the effective implementation and use of EMRs. PMID- 16046563 TI - Increasing capacity for innovation in bureaucratic primary care organizations: a whole system participatory action research project. AB - PURPOSE: We wanted to identify what organizational features support innovation in Primary Care Groups (PCGs). METHODS: Our study used a whole system participatory action research model. Four research teams provided complementary insights. Four case study PCGs were analyzed. Two had an intervention to help local facilitators reflect on their work. Data included 70 key informant interviews, observations of clinical governance interventions and committee meetings, analysis of written materials, surveys and telephone interviews of London Primary Care Organizations, interviews with 20 nurses, and interviews with 6 finance directors. A broad range of stakeholders reviewed data at annual conferences and formed conclusions about trustworthy principles. Sequential research phases were refocused in the light of these conclusions and in response to the changing political context. RESULTS: Five features were associated with increased organizational capacity for innovation: (1) clear structures and a vision for corporate and clinical governance; (2) multiple opportunities for people to reflect and learn at all levels of the organization, and connections between these "learning spaces"; (3) both clinicians and managers in leadership roles that encourage participation; (4) the right timing for an initiative and its adaptation to the local context; and (5) external facilitation that provides opportunities for people to make sense of their experiences. Low morale was commonly attributed to 3 features: (1) overwhelming pace of reform, (2) inadequate staff experience and supportive infrastructure, and (3) financial deficits. CONCLUSIONS: These features together may support innovation in other primary care bureaucracies. The research methodology enabled people from different backgrounds to make sense of diverse research insights. PMID- 16046564 TI - Stories from frequent attenders: a qualitative study in primary care. AB - PURPOSE: Patients who make frequent office visits (frequent attenders) in primary care are often considered a major burden on resources, yet we know little about their perceptions and expectations. We wanted to explore how these patients viewed their rates of consultation, what they expected from the consultation, and how they perceived their relationship with the primary health care team. METHODS: Using a qualitative study design, we undertook in-depth semi-structured interviews with frequent attenders at 4 primary care practices of the Mersey Primary Care R&D Consortium in the North West of England. Participants were identified on the basis of office visits at least twice the mean standardized rate for 1 year and a medical assessment that these visits had no important clinical outcome. Interviews with 30 patients aged 24 to 81 years (18 men) were audiotaped and transcribed, and the text was methodically coded; data were analyzed by generating common themes. RESULTS: Participants were unable or unwilling to quantify their consultation rates. Despite the assertion by many participants that family doctors are caring, authority figures, there was an underlying tension between such perceptions and the apparent medical mismanagement of symptoms. Their expectations of the consultation were complex and included the presentation of old and new symptoms implicitly embedded within an illness framework. Gaining access to family doctors was generally perceived as problematic. CONCLUSION: The criteria held by family doctors and researchers regarding the appropriate rate of consultations in primary care may not be shared by patients who attend frequently. Such patients require family doctors to acknowledge their symptoms and to provide reassurance. PMID- 16046565 TI - Predicting persistently high primary care use. AB - PURPOSE: We wanted to identify risk factors for persistently high use of primary care. METHODS: We analyzed outpatient office visits to practitioners in family medicine, general internal medicine, general pediatrics, and obstetrics for 1997 1999 among patients in a small Midwestern city covered by a fee-for-service insurance plan with no co-payments for physician visits and no requirement for referral to specialty care. Logistic regression was used to predict which patients with 10 or more primary care visits in 1997 would repeat high use in 1998 based on demographic and diagnostic categories (adjusted clinical groups [ACGs]). A confirmatory data set (high primary care use in 1998 persistent into 1999) was used to evaluate the model. RESULTS: Two percent of the 54,074 patients had 10 or more primary care visits in 1997, and of these, almost 19% had 10 or more visits in the next year. Among adults, 4 ambulatory diagnosis groups (ADGs) were simultaneously positive predictors of repeated high primary care visits: unstable chronic medical conditions, see and reassure conditions, minor time limited psychosocial conditions, and minor signs and symptoms. Meanwhile, pregnancy was negatively associated. The area under the receiver operating characteristic (ROC) curve was 0.794 for adults in the developmental data set and 0.752 in the confirmatory data set, indicating a moderately accurate assessment. A satisfactory model was not developed for pediatric patients. CONCLUSIONS: Many persistently high primary care users appear to be overserviced but underserved, with underlying problems not addressed by a medical approach. Some may benefit from psychosocial support, whereas others may be good candidates for disease management interventions. PMID- 16046566 TI - Do patients treated with dignity report higher satisfaction, adherence, and receipt of preventive care? AB - PURPOSE: Although involving patients in their own health care is known to be associated with improved outcomes, this study was conducted to determine whether respecting persons more broadly, such as treating them with dignity, has additional positive effects. METHODS: Using data from the Commonwealth Fund 2001 Health Care Quality Survey of 6,722 adults living in the United States, we performed survey-weighted logistic regression analysis to evaluate independent associations between 2 measures of respect (involvement in decisions and treatment with dignity) and patient outcomes (satisfaction, adherence, and receipt of optimal preventive care). Then we calculated adjusted probabilities of these outcomes and performed stratified analyses to examine results across racial/ethnic groups. RESULTS: After adjustment for respondents' demographic characteristics, the probability of reporting a high level of satisfaction was higher for those treated with dignity vs not treated with dignity (0.70 vs 0.38, P < .001) and for those involved in, versus not involved in, decisions (0.70 vs 0.39, P < .001). These associations were consistent across all racial/ethnic groups. Being involved in decisions was significantly associated with adherence for whites, whereas being treated with dignity was significantly associated with adherence for racial/ethnic minorities. The probability of receiving optimal preventive care was marginally greater for those treated with dignity (0.68 vs 0.63, P = .054), but did not differ with respect to involvement in decisions (0.67 vs 0.67, P = .95). CONCLUSIONS: Being treated with dignity and being involved in decisions are independently associated with positive outcomes. Although involving patients in decisions is an important part of respecting patient autonomy, it is also important to respect patients more broadly by treating them with dignity. PMID- 16046567 TI - Trust in one's physician: the role of ethnic match, autonomy, acculturation, and religiosity among Japanese and Japanese Americans. AB - PURPOSE: Trust is a cornerstone of the physician-patient relationship. We investigated the relation of patient characteristics, religiosity, acculturation, physician ethnicity, and insurance-mandated physician change to levels of trust in Japanese American and Japanese patients. METHODS: A self-administered, cross sectional questionnaire in English and Japanese (completed in the language of their choice) was given to community-based samples of 539 English-speaking Japanese Americans, 340 Japanese-speaking Japanese Americans, and 304 Japanese living in Japan. RESULTS: Eighty-seven percent of English-speaking Japanese Americans, 93% of Japanese-speaking Japanese Americans, and 58% of Japanese living in Japan responded to trust items and reported mean trust scores of 83, 80, and 68, respectively, on a scale ranging from 0 to 100. In multivariate analyses, English-speaking and Japanese-speaking Japanese American respondents reported more trust than Japanese respondents living in Japan (P values <.001). Greater religiosity (P <.001), less desire for autonomy (P <.001), and physician patient relationships of longer duration (P <.001) were related to increased trust. Among Japanese Americans, more acculturated respondents reported more trust (P <.001), and Japanese physicians were trusted more than physicians of another ethnicity. Among respondents prompted to change physicians because of insurance coverage, the 48% who did not want to switch reported less trust in their current physician than in their former physician (mean score of 82 vs 89, P <.001). CONCLUSIONS: Religiosity, autonomy preference, and acculturation were strongly related to trust in one's physician among the Japanese American and Japanese samples studied and may provide avenues to enhance the physician-patient relationship. The strong relationship of trust with patient-physician ethnic match and the loss of trust when patients, in retrospect, report leaving a preferred physician suggest unintended consequences to patients not able to continue with their preferred physicians. PMID- 16046568 TI - The doctor who cried: a qualitative study about the doctor's vulnerability. AB - PURPOSE: We wanted to explore those clinical events when doctors had exposed their vulnerability toward patients in a potentially beneficial way. METHODS: We undertook a qualitative study based on memory work, a structured approach to transform memories into written texts. Study participants were 9 members of a research group who had known each other a couple of years. They were asked in advance to recall a clinical event during which vulnerability was perceived and exposed in a way appreciated positively by the patient. During a group meeting, participants wrote their individual memory stories recalling these events, and the subsequent group discussion was audiotaped, transcribed, and analyzed using a phenomenological approach, applying specific linguistic cues to reveal points of special interest. The main outcome measure was the vulnerability expressed by practitioners. RESULTS: Vulnerability had been experienced and exposed by the participants on several occasions during which the patients had confirmed its potentially beneficial effect. All reported events could be interpreted as different ways of personal disclosure toward the patient. We identified two kinds of disclosure: spontaneously appearing emotions and considered sharing of experiences. CONCLUSION: A spontaneous exposure of emotions from the doctor may help the patient, and sharing personal experiences may lead to constructive interaction. We need to know more about when and how personal disclosure and other aspects of vulnerability exposed by the doctor are experienced as beneficial by the patient. PMID- 16046569 TI - Continuity of care: is the personal doctor still important? A survey of general practitioners and family physicians in England and Wales, the United States, and The Netherlands. AB - PURPOSE: We determined the reported value general practitioners/family physicians in 3 different health care systems place on the various types of continuity of care. METHODS: We conducted a postal questionnaire survey in England and Wales, the United States, and The Netherlands. The participants were 1,523 general practitioners/family physicians (568 from England and Wales, 453 from the United States and 502 from The Netherlands). Our main outcome measures were the perceived importance of the types of continuity of care and doctor or practice characteristics that may influence attitudes toward personal continuity of care. RESULTS: The response rates were England and Wales 60% (568/946), United States 47% (453/963) and Netherlands 76% (502/660). The doctors in all 3 countries felt strongly that personal continuity remained an important aspect of good-quality care to their patients. Within a given health care system, doctors' personal and practice characteristics explained only a small part of the variance in attitudes toward the provision of personal continuity of care (England and Wales and The Netherlands r2 = 0.04, United States r2 = 0.01). The doctors in all 3 countries felt that they were currently able to provide all 3 types of continuity of care, although doctors in England and Wales were least positive about the provision of informational and management continuity across the primary-secondary care divide. CONCLUSIONS: General practitioners/family physicians from 3 differing health care systems all place high value on being able to provide personal continuity of care to patients. Personal continuity of care remains a core value of general practice/family medicine and should be taken account of by policy makers when redesigning health care systems. PMID- 16046570 TI - Enhance your team-based qualitative research. AB - PURPOSE: Qualitative research projects often involve the collaborative efforts of a research team. Challenges inherent in teamwork include changes in membership and differences in analytical style, philosophy, training, experience, and skill. This article discusses teamwork issues and tools and techniques used to improve team-based qualitative research. METHODS: We drew on our experiences in working on numerous projects of varying, size, duration, and purpose. Through trials of different tools and techniques, expert consultation, and review of the literature, we learned to improve how we build teams, manage information, and disseminate results. RESULTS: Attention given to team members and team processes is as important as choosing appropriate analytical tools and techniques. Attentive team leadership, commitment to early and regular team meetings, and discussion of roles, responsibilities, and expectations all help build more effective teams and establish clear norms. As data are collected and analyzed, it is important to anticipate potential problems from differing skills and styles, and how information and files are managed. Discuss analytical preferences and biases and set clear guidelines and practices for how data will be analyzed and handled. As emerging ideas and findings disperse across team members, common tools (such as summary forms and data grids), coding conventions, intermediate goals or products, and regular documentation help capture essential ideas and insights. CONCLUSIONS: In a team setting, little should be left to chance. This article identifies ways to improve team-based qualitative research with more a considered and systematic approach. Qualitative researchers will benefit from further examination and discussion of effective, field-tested, team-based strategies. PMID- 16046571 TI - Making time to write? AB - Clinicians are busy people who must develop strategies for making time to write. With many obligations, their first strategy may be to try to clear the decks to make way for the current project. When writing takes on a more constant challenge, medical writers find themselves wedging writing in between myriad other tasks and commitments. Ultimately, committed writers, with effort and help, find a way to schedule writing as another essential activity in their lives. PMID- 16046572 TI - On this day of mothers and sons. AB - Telling a pregnant woman and her family about a fetal demise is one of the greatest challenges for maternity care clinicians. In this essay, the author reflects on such an encounter in her work as a community health center clinician, maternity care teacher, and mother herself. PMID- 16046574 TI - Dietrich receives Curtis G. Hames Research Award, call for new model papers. PMID- 16046579 TI - Guideline showcases AAFP's commitment to evidence-based, patient-centered care. PMID- 16046580 TI - Vascular endothelial growth factor Trap suppresses ovarian function at all stages of the luteal phase in the macaque. AB - CONTEXT: Fertility is dependent on a functioning corpus luteum, the formation of which is associated with intense angiogenesis. The role of angiogenic factors, such as vascular endothelial growth factor (VEGF), in luteal function has yet to be defined in primates. OBJECTIVE: The objective of this study was to determine effects of inhibiting VEGF by a VEGF Trap, a receptor-based inhibitor, administered at the early or midluteal phase, on pituitary-ovarian function. DESIGN: Effects of a single injection of VEGF Trap at three doses in the early luteal phase or a single dose in the midluteal phase were investigated and compared with control cycles. SETTING: This work was conducted in the Primate Unit. PARTICIPANTS: Eleven stump-tailed macaques with regular ovulatory cycles participated in this study. VEGF Trap was well tolerated, and all completed the study. INTERVENTIONS: A single injection of VEGF Trap at a dose of 4, 1, or 0.25 mg/kg was administered in the early luteal phase or 1 mg/kg in the midluteal phase. Controls received vehicle or the constant region of human IgG. MAIN OUTCOME MEASURES: Changes in serum concentrations of progesterone, estradiol, LH, FSH, inhibin A, VEGF Trap, and menstrual bleeding were the main outcome measures. RESULTS: Early luteal treatment caused a significant attenuation of the normal serum progesterone and estradiol concentrations, followed by a marked increase in LH and FSH. Inhibin A was not significantly reduced. After 1- and 4-mg/kg doses, progesterone remained suppressed throughout the luteal phase, and premature menstruation occurred; whereas the response to the 0.25-mg/kg dose was transitory, and menstruation was at the normal time. Midluteal treatment also resulted in a significant suppression of progesterone secretion. CONCLUSIONS: VEGF is essential for both the development and maintenance of luteal function. PMID- 16046581 TI - Genetic variation in the hypoxia-inducible factor-1alpha gene is associated with type 2 diabetes in Japanese. AB - CONTEXT AND OBJECTIVE: Vascular endothelial growth factor plays a critical role both in neovascularization of proliferative diabetic retinopathy and in angiogenesis of islets in the pancreatic developmental stage in determining beta cell mass and properties. Vascular endothelial growth factor mRNA levels increase as a result of increased transcriptional activation, mediated predominantly by hypoxia-inducible factor-1 alpha (HIF-1alpha) in response to hypoxia. DESIGN AND PATIENTS: In this study, we examined all regions of the HIF-1alpha to detect single-nucleotide polymorphisms (SNPs), evaluated the pattern of linkage disequilibrium to analyze haplotypes, and performed association studies in Japanese type 2 diabetes patients with or without retinopathy. RESULTS: A total of 35 SNPs were found in the gene, 27 of which were reported previously and eight of which were novel. Three of the 35 SNPs were located in coding regions, one in exon 2 (S28Y), and the others in exon 12 (P582S, A588T). The P582S HIF-1alpha mutation was associated with type 2 diabetes (P = 0.0028) by a consistently higher level of transcriptional activity than wild type, especially under hypoxic condition (P = 0.012), but no association with retinopathy was detected. CONCLUSION: This is the first report that HIF-1alpha is associated with the occurrence of type 2 diabetes and suggests that the P582S HIF-1alpha mutation should be assessed in larger studies as a risk factor for type 2 diabetes. PMID- 16046582 TI - Comparative assessment in young and elderly men of the gonadotropin response to aromatase inhibition. AB - CONTEXT: Aging in men is associated with a decline in serum testosterone (T) levels. OBJECTIVE: Our objective was to assess whether decreased T in aging might result from increased estradiol (E2) negative feedback on gonadotropin secretion. DESIGN AND SETTING: We conducted a comparative intervention study (2004) in the Outpatient Endocrinology Clinic, Ghent University Hospital. PARTICIPANTS: Participants included healthy young and elderly men (n = 10 vs. 10). INTERVENTIONS: We used placebo and letrozole (2.5 mg/d) for 28 d, separated by 2 wk washout. MAIN OUTCOME MEASURES: We assessed changes in serum levels of free E2, LH, and FSH, free T, SHBG, and gonadotropins response to an i.v. 2.5-microg GnRH bolus. RESULTS: As assessed after 28 d of treatment, letrozole lowered E2 by 46% in the young men (P = 0.002) and 62% in the elderly men (P < 0.001). In both age groups, letrozole, but not placebo, significantly increased LH levels (339 and 323% in the young and the elderly, respectively) and T (146 and 99%, respectively) (P value of young vs. elderly was not significant). Under letrozole, peak LH response to GnRH was 152 and 52% increase from baseline in young and older men, respectively (P = 0.01). CONCLUSIONS: Aromatase inhibition markedly increased basal LH and T levels and the LH response to GnRH in both young and elderly men. The observation of similar to greater LH responses in the young compared with the elderly does not support the hypothesis that increased restraining of LH secretion by endogenous estrogens is instrumental in age related decline of Leydig cell function. PMID- 16046583 TI - Response of matrix metalloproteinases and tissue inhibitors of metalloproteinases messenger ribonucleic acids to ovarian steroids in human endometrial explants mimics their gene- and phase-specific differential control in vivo. AB - CONTEXT: Cyclic remodeling and breakdown of the extracellular matrix, a unique feature of the human endometrium, depends on matrix metalloproteinases (MMPs). These enzymes are globally controlled by estradiol and progesterone or their withdrawal, but various MMPs and their tissue inhibitors (TIMPs) show distinct responses. OBJECTIVE AND DESIGN: To clarify the role of ovarian steroids in the differential regulation of MMP-1, MMP-3, MMP-7, MMP-8, MMP-10, TIMP-1, TIMP-2, and TIMP-3 mRNAs, we compared their variations in the cycling endometrium in vivo with their response to hormone addition or withdrawal in corresponding explants. RESULTS: Different patterns were identified in vivo according to the time frame (secretory vs. perimenstrual increase), sharpness (peak vs. progressive increase or decrease), and magnitude of the changes. In vivo ratios between early/midsecretory and perimenstrual phases ranged from more than 1000 (MMP-1, MMP-3, and MMP-10) to less than 10 (TIMPs). Differential response to ovarian steroids of the various MMPs and TIMPs mRNAs tested in cultured explants matched the same ranking and varied according to the phase at sampling. Remarkably, ovarian steroids repressed MMPs and TIMP-1 and TIMP-2 but, in secretory explants, increased TIMP-3 mRNA. Finally, in situ hybridization evidenced the major contribution of fibroblasts to the increase in MMP-8 mRNA at menstruation or in explants cultured without hormones. CONCLUSIONS: Both phase- and gene-specific modulators finely tune in space, time, and amplitude the global control of MMPs and TIMPs mRNAs by estradiol and progesterone in the cycling human endometrium. PMID- 16046584 TI - Menopausal hormone therapy and irregular endometrial bleeding: a potential role for uterine natural killer cells? AB - CONTEXT: Irregular bleeding affects many users of combined menopausal hormone therapy (HT) and commonly leads to invasive and expensive investigations to exclude underlying malignancy. In most cases no abnormality is found. OBJECTIVE: The main objective of this study was to explore the role of uterine natural killer (uNK) cells and their regulatory cytokine IL-15 in irregular bleeding in HT users. DESIGN: This was a prospective observational study conducted between 2002 and 2004. SETTING: The study was conducted in a tertiary referral menopause clinic at King Edward Memorial Hospital, Western Australia. PATIENTS: Patients included 117 postmenopausal women taking combined HT. INTERVENTIONS: Outpatient endometrial biopsies were taken during and outside bleeding episodes. MAIN OUTCOME MEASURES: The relationship between endometrial uNK cells (CD56+) and bleeding patterns was measured. We also addressed the impact of HT exposure on uNK cell populations, the relationship between endometrial IL-15 expression and uNK cell populations, and killer Ig like receptor genotype in subjects with irregular bleeding. RESULTS: Endometrial CD56+ uNK cells were significantly increased in biopsies obtained during bleeding episodes (P < 0.001), compared with HT users with no bleeding. The highest level of IL-15 expression was also seen in biopsies taken during bleeding. No clear relationship between killer Ig like receptor genotype and bleeding on HT was observed. CONCLUSIONS: Little is known about the mechanisms underlying irregular bleeding in HT users. This is the first report of uNK cells and their association with regulating cytokines in postmenopausal endometrium and demonstrates a possible mechanism by which HT may induce irregular bleeding. PMID- 16046585 TI - Changes in plasma concentrations of osteoprotegerin before and after levothyroxine replacement therapy in hypothyroid patients. AB - CONTEXT: Recent study has shown that overt hypothyroidism (oHT) is associated with increased plasma osteoprotegerin (OPG) levels. OBJECTIVE: Our objective was to examine the plasma OPG level alteration before and after levothyroxine (L-T4) treatment in oHT and subclinical hypothyroidism (sHT). PATIENTS: The study subjects included oHT and sHT patients and healthy individuals (20 subjects in each group). METHODS: All patients were given L-T4 therapy to maintain a euthyroid state. Plasma OPG concentration was measured in duplicate by a sandwich ELISA. RESULTS: Plasma OPG levels in oHT and sHT before treatment were significantly higher than levels in controls (P < 0.01). After normalization of thyroid function, OPG levels in both groups decreased markedly (P < 0.01). The absolute changes in OPG showed a significant positive correlation with the changes in TSH (P < 0.05) and negative correlation with the changes in endothelium-dependent arterial dilation (P < 0.01) in hypothyroid patients during the course of treatment. CONCLUSION: OPG may be involved in the development of vascular dysfunction in hypothyroid patients. PMID- 16046586 TI - Cotreatment of acromegaly with a somatostatin analog and a growth hormone receptor antagonist. AB - CONTEXT: Pegvisomant is a GH receptor antagonist that blocks the peripheral actions of GH in acromegaly. Pegvisomant, in contrast to somatostatin (SMS) analogs, does not suppress the activity of the GH-producing adenoma. OBJECTIVE: We assessed the effects of cotreatment with pegvisomant and SMS in acromegaly on GH secretion, IGF-I levels, and glucose tolerance. DESIGN, PATIENTS, AND INTERVENTIONS: Eleven patients with persistent disease despite previous therapy underwent the following fixed treatment algorithm: 1) on SMS therapy, 2) off therapy for 2 months, 3) 6-wk treatment with 10 mg/d pegvisomant, 4) 6-wk treatment with 15 mg/d pegvisomant, and 5) 3-month treatment with 15 mg pegvisomant plus SMS. Blood was sampled in the fasting state and during an oral glucose tolerance test. RESULTS: Total serum IGF-I levels (micrograms per liter) decreased after pegvisomant, but the lowest levels were obtained with cotreatment [458 +/- 67 (SMS), 562 +/- 78 (active), 376 +/- 51 (10 mg), 269 (15 mg), 195 +/- 24 (combined) (P < 0.0001)]. Free and bioactive IGF-I changed in a similar pattern. Steady-state pegvisomant levels (micrograms per liter) were obtained, but SMS cotreatment increased pegvisomant levels by 20% (P = 0.02) [2631 +/- 616 (10 mg), 6536 +/- 1413 (15 mg), 8030 +/- 1914 (combined)]. Pegvisomant increased endogenous GH levels (micrograms per liter), which was countered by SMS cotreatment [5.1 +/- 1.3 (SMS), 8.9 +/- 2.9 (active), 14.6 +/- 4.9 (10 mg), 19.7 +/- 6.5 (15 mg), 11.8 +/- 2.8 (combined) (P < 0.01)]. Plasma glucose levels (millimoles per liter) were highest during SMS and lowest during pegvisomant 15 mg [2-h oral glucose tolerance test: 10.3 +/- 0.7 (SMS), 8.9 +/- 0.7 (active), 7.2 +/- 0.7 (10 mg), 6.5 +/- 0.5 (15 mg), 8.0 +/- 0.8 (combined) (P = 0.02)]. CONCLUSIONS: Dual blockade of the GH axis with pegvisomant and a SMS analog is feasible in acromegaly. PMID- 16046587 TI - Effect of smoking on uric acid and other metabolic markers throughout normal pregnancy. AB - OBJECTIVES: Smoking, pregnancy, and preeclampsia are all associated with changes in markers of the metabolic syndrome. Several markers are increased in all three conditions. However, smoking is negatively associated with preeclampsia, and therefore some markers would be expected to behave differently in smokers during pregnancy. We compared several metabolic markers of the metabolic syndrome in healthy primigravid smokers and nonsmokers over normal pregnancy to explore mechanisms for the reduced risk of preeclampsia in smokers. STUDY DESIGN: Plasma was obtained from 63 women throughout pregnancy who delivered at term. Smoking status was determined by urinary cotinine concentrations measured by HPLC. Uric acid, creatinine, free fatty acids, triglycerides, and total cholesterol were measured with diagnostic kits. Data were analyzed by repeated-measures ANOVA. RESULTS: The smoking groups were not different by delivery gestational age, maternal age, body mass index, or race. Uric acid, cholesterol, and triglyceride concentrations increased during pregnancy (significant for time, P < 0.0001). Mean uric acid and creatinine concentrations were different by smoking status (P < 0.001 and P = 0.046). Nonsmokers had the lowest concentrations of uric acid, and women who quit smoking had the highest concentrations. Uric acid concentrations remained significantly different controlling for serum creatinine CONCLUSIONS: Women have changes in markers of the metabolic syndrome during pregnancy, and uric acid is further influenced by smoking. The difference in uric acid concentrations by smoking status may be secondary to increased production through the xanthine oxidase pathway but is not simply a result of altered glomerular function because the association persists after controlling for creatinine. PMID- 16046588 TI - 21-Hydroxylase and 11beta-hydroxylase mutations in Romanian patients with classic congenital adrenal hyperplasia. AB - CONTEXT: Congenital adrenal hyperplasia (CAH) comprises autosomal recessive disorders mainly due to defects in the 21-hydroxylase (CYP21) gene. OBJECTIVE: The study aimed to perform molecular characterization in 43 Romanian patients with classical CAH forms diagnosed at the Center for Genetic Diseases of the Pediatric Clinic/University Cluj (38 with 21-hydroxylase deficiency, five with 11beta-hydroxylase deficiency), to determine the frequency of mutations in the CYP21A2 gene and attempt a genotype-phenotype correlation in patients with 21 hydroxylase deficiency. DESIGN: Molecular analysis was performed by direct sequencing of PCR amplified products of the CYP21A2 and CYP11B1 genes. RESULTS: The most frequent mutation in Romanian patients with 21-hydroxylase deficiency was I2G (43.9%), followed by deletions and large conversions (16.7%), I172N and the triple mutation (P30L+I2G+del8bp), accounting for 12.1% each, P30L (7.6%) and R356W (1.5%). Genotypes were categorized in three mutation groups (0, A, and B), according to their predicted functional consequences, and compared with clinical phenotype. Positive predictive values were 100, 75, and 100% for groups 0, A, and B, respectively. Overall genotype-phenotype correlation was 87.88%. In the five patients with 11beta-hydroxylase deficiency, the following homozygous mutations were identified: T318R in two related patients; R448H in two unrelated patients; and P94L, a new, yet-undescribed mutation. CONCLUSION: The present study is the first countrywide report of mutational analysis in a Romanian patient population with 21-hydroxylase deficiency. Molecular diagnosis was performed in a small number of CAH patients proved not to suffer from 21-hydroxylase deficiency but from 11beta-hydroxylase deficiency, and a new mutation was identified. PMID- 16046589 TI - Direct in vitro evidence of extracellular Ca2+-induced amino-terminal truncation of human parathyroid hormone (1-84) by human parathyroid cells. AB - CONTEXT: Although serum calcium (Ca2+) concentration regulates the generation of amino-terminally (N-terminally) truncated forms of human PTH (hPTH) degraded from (1-84)hPTH, no studies have yet reported whether the parathyroid gland itself is responsible for this process. OBJECTIVE: Our objective was to determine the site of N-terminal truncation and its roles in PTH metabolism in parathyroid cells in vitro. METHODS: The effect of extracellular Ca2+ concentration was examined on N terminal truncation in primary cultured parathyroid cells. The parathyroid glands were obtained from the patients with primary and uremia-associated secondary hyperparathyroidisms who underwent therapeutic parathyroidectomies. RESULTS: The N-terminally truncated fragments were detectable with commercially available intact PTH (I-PTH) assays, but not with the bio-intact PTH (Bio-PTH) assay, which detected only the (1-84)hPTH. HPLC revealed that generation of N-terminally truncated fragments detectable by I-PTH increased with extracellular Ca2+ concentration. Suppression of PTH secretion by increasing the extracellular Ca2+ concentration was more evident with the Bio-PTH assay than with the I-PTH assay for both cultured parathyroid cells prepared from parathyroid adenomas and uremia associated secondary hyperparathyroidism. The Bio-PTH/I-PTH ratio, which is the ratio of (1-84)hPTH to the sum of (1-84)hPTH and N-terminally truncated fragments, decreased in response to increases in extracellular Ca2+. CONCLUSIONS: These findings suggest that the N-terminal truncation is regulated by extracellular Ca2+ concentration and works to suppress the generation of (1 84)hPTH in parathyroid cells. PMID- 16046590 TI - Overweight women with polycystic ovary syndrome have evidence of subclinical cardiovascular disease. AB - CONTEXT: Polycystic ovary syndrome (PCOS) is associated with insulin resistance (IR) and the metabolic syndrome. There are no adequate data demonstrating significantly increased cardiovascular disease (CVD) mortality. In the absence of clinical outcome studies, surrogate markers of early CVD can provide insight into early CVD. OBJECTIVE: The aim of this study was to clarify whether overweight women with PCOS have an increased prevalence of cardiovascular risk factors and early CVD, compared with age- and body mass index-matched controls, to determine the contribution of PCOS per se to CVD status. DESIGN AND PATIENTS: This was a case control study of 100 overweight women with PCOS and 20 subjects of similar body mass index and age. MAIN OUTCOME MEASURES: Noninvasive markers of early CVD [carotid intimal media thickness, pulse wave velocity (PWV), and brachial arterial flow-mediated vasodilation] were measured. Metabolic parameters studied included insulin, glucose, C-reactive protein, lipids, and androgens. RESULTS: Subjects with PCOS had elevated testosterone (2.5 +/- 0.2 vs. 1.3 +/- 0.1 nmol/liter), dehydroepiandrosterone sulfate (4.9 +/- 0.3 vs. 3.6 +/- 0.4 mmol/liter), fasting insulin (19.6 +/- 1.4 vs. 6.8 +/- 0.8 microU/ml), and homeostasis model assessment of IR (4.1 +/- 0.3 vs. 1.3 +/- 0.2), compared with controls. In addition, those with PCOS had elevated cholesterol (5.1 +/- 0.1 vs. 4.6 +/- 0.2 mmol/liter) and triglycerides (1.4 +/- 0.1 vs. 0.9 +/- 0.1 mmol/liter), whereas there were no differences in either C-reactive protein or 24 h ambulatory blood pressure parameters. Subjects with PCOS also had increased arterial stiffness (PWV, 7.4 +/- 0.1 vs. 6.6 +/- 0.2 m/sec) and endothelial dysfunction (flow-mediated vasodilation, 9.8 +/- 0.4 vs. 13.3 +/- 0.9), compared with controls. There was no difference in mean intimal media thickness between the groups. Stepwise regression in PCOS subjects showed that IR and lipids were independent predictors of PWV. CONCLUSION: Overweight women with PCOS have increased cardiovascular risk factors and evidence of early CVD, compared with weight-matched controls, potentially related to IR. PMID- 16046591 TI - Repeated [131I]metaiodobenzylguanidine therapy in two patients with malignant pheochromocytoma. AB - CONTEXT: Approximately 10% of pheochromocytomas are malignant with a 5-yr survival rate of less than 40%. Promising results have been published on single high-dosage [131I]metaiodobenzylguanidine ([131I]MIBG) treatment for malignant pheochromocytoma. We present our experience with multiple intermediate-dosage [131I]MIBG therapy instead of single high-dosage therapy. SETTING AND PATIENTS: The study took place at University Medical Centers and included two patients (one male, 36 yr of age, and one female, 43 yr of age) with widely spread metastatic pheochromocytoma and bad prognosis because of liver and lung metastases. INTERVENTIONS: Instead of a single high dosage, these two patients were treated with multiple intermediate dosages of [131I]MIBG. The first patient received 37 GBq (1 Ci) [131I]MIBG in five sessions [7400 MBq (200 mCi) each; interval range, 2-11 months]; the second patient received 66.6 GBq (1.8 Ci) [131I]MIBG in 12 sessions [5550 MBq (150 mCi) each; interval range, 2-14 months]. OUTCOME MEASURES: We measured efficacy, toxicity, and survival. RESULTS: Both patients had a complete symptomatic response and a partial tumor volume response. The first patient had a partial biochemical response, the second a complete biochemical response. In both cases, toxicity has been confined to nausea during treatment. Hematological toxicity was minimal, and both patients stayed euthyroid. The survival (so far) of these patients was 5 yr (clinical case 1) and 16 yr (clinical case 2) after initial diagnosis. CONCLUSIONS: Repeated intermediate-dosage [131I]MIBG treatment appears to be a reliable and well tolerated radionuclide therapy and might be a useful adjunct in patients with malignant pheochromocytoma, providing longstanding palliation and prolonged survival. PMID- 16046593 TI - The genesis of HCFA. PMID- 16046594 TI - Turning Medicare and Medicaid into health programs: the role of organizational culture. PMID- 16046595 TI - Medicare, medicaid, and health care quality. PMID- 16046596 TI - The twin policy challenges of Medicare physician payment and Medicaid. PMID- 16046597 TI - My life at HCFA: a true parable. PMID- 16046598 TI - What does it take to run Medicare and Medicaid? PMID- 16046599 TI - Policy high points: Medicare and Medicaid in the new millennium. PMID- 16046600 TI - Occupational exposure to glycol ethers and ovarian function. PMID- 16046601 TI - Self report and GIS based modelling as indicators of air pollution exposure: is there a gold standard? PMID- 16046602 TI - Prolonged menstrual cycles in female workers exposed to ethylene glycol ethers in the semiconductor manufacturing industry. AB - BACKGROUND: It has been shown that female workers exposed to ethylene glycol ethers (EGEs) in the semiconductor industry have higher risks of spontaneous abortion, subfertility, and menstrual disturbances, and prolonged waiting time to pregnancy. AIMS: To examine whether EGEs or other chemicals are associated with long menstrual cycles in female workers in the semiconductor manufacturing industry. METHODS: Cross-sectional questionnaire survey during the annual health examination at a wafer manufacturing company in Taiwan in 1997. A three tiered exposure-assessment strategy was used to analyse the risk. A short menstrual cycle was defined to be a cycle less than 24 days and a long cycle to be more than 35 days. RESULTS: There were 606 valid questionnaires from 473 workers in fabrication jobs and 133 in non-fabrication areas. Long menstrual cycles were associated with workers in fabrication areas compared to those in non-fabrication areas. Using workers in non-fabrication areas as referents, workers in photolithography and diffusion areas had higher risks for long menstrual cycles. Workers exposed to EGEs and isopropanol, and hydrofluoric acid, isopropanol, and phosphorous compounds also showed increased risks of a long menstrual cycle. CONCLUSIONS: Exposure to multiple chemicals, including EGEs in photolithography, might be associated with long menstrual cycles, and may play an important role in a prolonged time to pregnancy in the wafer manufacturing industry; however, the prevalence in the design, possible exposure misclassification, and chance should be considered. PMID- 16046603 TI - Exposure to traffic related air pollutants: self reported traffic intensity versus GIS modelled exposure. AB - BACKGROUND: In epidemiological studies of the potential health effects of traffic related air pollution, self reported traffic intensity is a commonly used, but rarely validated, exposure variable. METHODS: As part of a study on the impact of Traffic Related Air Pollution on Childhood Asthma (TRAPCA), data from 2633 and 673 infants from the Dutch and the German-Munich cohorts, respectively, were available. Parents subjectively assessed traffic intensity at the home address. Objective exposures were estimated by a combination of spatial air pollution measurements and geographic information system (GIS) based modelling using an identical method for both cohorts. RESULTS: The agreement rates between self reported and GIS modelled exposure--accumulated over the three strata of self assessed traffic intensity--were 55-58% for PM(2.5), filter absorbance (PM(2.5) abs), and nitrogen dioxide in Munich and 39-40% in the Netherlands. Of the self reported low traffic exposed group, 71-73% in Munich and 45-47% in the Netherlands had low modelled exposure to these three air pollutants. Of the self assessed high exposed subgroups in Munich (15% of the total population) and the Netherlands (22% of the total population), only 22-33% and 30-32% respectively had high modelled exposure to the three air pollutants. The subjective assessments tend to overestimate the modelled estimates for PM(2.5) and NO2 in both study areas. When analysis was restricted to the portion of the Dutch cohort living in non-urban areas, the agreement rates were even lower. CONCLUSIONS: Self reported and modelled assessment of exposure to air pollutants are only weakly associated. PMID- 16046604 TI - Impact of ambient air pollution on birth weight in Sydney, Australia. AB - BACKGROUND: Studies in Asia, Europe, and the Americas have provided evidence that ambient air pollution may have an adverse effect on birth weight, although results are not consistent. METHODS: Average exposure during pregnancy to five common air pollutants was estimated for births in metropolitan Sydney between 1998 and 2000. The effects of pollutant exposure in the first, second, and third trimesters of pregnancy on risk of "small for gestational age" (SGA), and of pollutant exposure during pregnancy on birth weight were examined. RESULTS: There were 138,056 singleton births in Sydney between 1998 and 2000; 9.7% of babies (13,402) were classified as SGA. Air pollution levels in Sydney were found to be quite low. In linear regression models carbon monoxide and nitrogen dioxide concentrations in the second and third trimesters had a statistically significant adverse effect on birth weight. For a 1 part per million increase in mean carbon monoxide levels a reduction of 7 (95% CI -5 to 19) to 29 (95% CI 7 to 51) grams in birth weight was estimated. For a 1 part per billion increase in mean nitrogen dioxide levels a reduction of 1 (95% CI 0 to 2) to 34 (95% CI 24 to 43) grams in birth weight was estimated. Particulate matter (diameter less than ten microns) in the second trimester had a small statistically significant adverse effect on birth weight. For a 1 microgram per cubic metre increase in mean particulate matter levels a reduction of 4 grams (95% CI 3 to 6) in birth weight was estimated. CONCLUSION: These findings of an association between carbon monoxide, nitrogen dioxide, and particulate matter, and reduction in birth weight should be corroborated by further study. PMID- 16046605 TI - The influence of occupational exposure to pesticides, polycyclic aromatic hydrocarbons, diesel exhaust, metal dust, metal fumes, and mineral oil on prostate cancer: a prospective cohort study. AB - AIMS: To investigate the relation between exposure to pesticides, polycyclic aromatic hydrocarbons (PAHs), diesel exhaust, metal dust, metal fumes, and mineral oil in relation to prostate cancer incidence in a large prospective study. METHODS: This cohort study was conducted among 58,279 men in the Netherlands. In September 1986, cohort members (55-69 years) completed a self administered questionnaire on potential cancer risk factors, including job history. Follow up for prostate cancer incidence was established by linkage to cancer registries until December 1995 (9.3 years of follow up). The analyses included 1386 cases of prostate cancer and 2335 subcohort members. A blinded case by-case expert exposure assessment was carried out to assign cases and subcohort members a cumulative probability of exposure for each potential carcinogenic exposure. RESULTS: In multivariate analyses there was a significant negative association for pesticides (RR 0.60; 95% CI 0.37 to 0.95) when comparing the highest tertile of exposure to pesticides with no exposure. No association was found for occupational exposure to PAHs (RR 0.75; 95% CI 0.42 to 1.31), diesel exhaust (RR 0.81; 95% CI 0.62 to 1.06), metal dust (RR 1.01; 95% CI 0.72 to 1.40), metal fumes (RR 1.11; 95% CI 0.80 to 1.54), or mineral oil (RR 0.99; 95% CI 0.66 to 1.48) when comparing the highest tertile of exposure with no exposure. In subgroup analysis, with respect to tumour invasiveness and morphology, null results were found for occupational exposure to pesticides, PAH, diesel exhaust, metal dust, metal fumes, and mineral oil. CONCLUSIONS: These results suggest a negative association between occupational exposure to pesticides and prostate cancer. For other carcinogenic exposures results suggest no association between occupational exposure to PAHs, diesel exhaust, metal dust, metal fumes, or mineral oil and prostate cancer. PMID- 16046606 TI - Perceptions of the impact of depression and anxiety and the medication for these conditions on safety in the workplace. AB - BACKGROUND: The number of people taking prescribed medication for anxiety and depression has increased greatly, but little is known of how this medication impacts on safety at work. AIMS: To examine the relation between anxiety and depression, prescribed medication, performance, and safety in the workplace. METHODS: The research involved nine focus groups with sufferers of anxiety and depression to investigate experiences of mental health problems and the impact of psychotropic drugs. A further three focus groups were conducted with staff in human resources, personnel, occupational health, and health and safety departments, to explore organisational perspectives. The sample comprised 74 individuals drawn from a wide range of occupational sectors. Finally, the results were presented to a panel of experts from occupational medicine, general practice, psychology, health and safety, and psychiatry, to consider the implications for practice. RESULTS: Workers reported that both the symptoms and the medication impaired work performance. Participants described accidents which they attributed to their condition or to the medication. Workers with responsibilities for others, such as teachers, healthcare workers, and managers appeared to present a particular safety risk. Healthcare workers believed that they placed themselves and their patients at risk when carrying out medical procedures. CONCLUSIONS: Respondents in this study felt that their symptoms of anxiety and depression and the medication they took to treat these conditions placed them at risk with respect to safety in the workplace. Drawing on the results, the authors outline areas for improvement in the management of mental health problems at work. PMID- 16046607 TI - The association between psychosocial characteristics at work and problem drinking: a cross-sectional study of men in three Eastern European urban populations. AB - BACKGROUND: Psychosocial factors at work are thought to influence health partly through health behaviours. AIMS: To examine the association between effort-reward imbalance and job control and several alcohol related measures in three eastern European populations. METHODS: A cross-sectional study was conducted in Novosibirsk (Russia), Krakow (Poland), and Karvina (Czech Republic). The participants completed a questionnaire that included effort-reward at work, job control, and a number of sociodemographic variables. Annual alcohol intake, annual number of drinking sessions, the mean dose of alcohol per drinking session, and binge drinking (> or =80 g of ethanol in one session at least once a week) were based on graduated frequencies in the questionnaire. Data were also available on problem drinking (> or =2 positive answers on CAGE questionnaire) and negative social consequences of drinking. All male participants in full employment (n = 694) were included in the present analyses. RESULTS: After controlling for age and centre, all indices of alcohol consumption and problem drinking were associated with the effort-reward ratio. Adjustment for material deprivation did not change the results but adjustment for depressive symptoms reduced the estimated effects. Job control was not associated with any of the alcohol related outcomes. CONCLUSIONS: The imbalance of effort-reward at work is associated with increased alcohol intake and problem drinking. The association appears to be partly mediated by depressive symptoms, which might be either an antecedent or a consequence of men's drinking behaviour. PMID- 16046608 TI - Investigating time patterns of variation in radiation cancer associations. AB - AIMS: In occupational settings, carcinogenic exposures are often repeated or protracted over time. The time pattern of exposure accrual may influence subsequent temporal patterns of cancer risk. The authors present several simple models that may be used to evaluate the influence of time since exposure or age at exposure on cancer incidence or mortality in an occupational cohort. METHODS: A cohort of 40,415 nuclear industry workers was identified via the Canadian National Dose Registry. Vital status and cause of death were ascertained through 1994. Associations between ionising radiation and mortality due to lung cancer, leukaemia, and cancers other than lung and leukaemia were quantified using conditional logistic regression models with risk sets constructed by incidence density sampling. A step function, a bilinear function, and a sigmoid function were used to evaluate temporal variation in exposure effects. RESULTS: Step and sigmoid functions were used to explore latency and morbidity periods. For analyses of lung cancer, leukaemia, and other cancers the best fitting models were obtained when exposure assignment was lagged by 13, 0, and 5 years, respectively. A bilinear function was used to evaluate whether exposure effects diminished with time since exposure. In analyses of lung cancer and leukaemia, there was evidence that radiation effects attenuated with protracted time since exposure. In analyses of age at exposure, there was evidence of variation in radiation mortality associations for analyses of lung cancer and leukaemia; discounting radiation doses accrued at younger ages (for example, 15-35 years) led to significant improvements in model fit. CONCLUSIONS: This paper illustrates empirical approaches to evaluating temporal variation in the effect of a protracted exposure on disease risk. PMID- 16046609 TI - Construction work and risk of occupational disability: a ten year follow up of 14,474 male workers. AB - AIMS: Most industrialised countries have public income maintenance programmes to protect workers in case of disability but studies addressing disability risk of specific professional groups are rare. The objective of this study was to establish a detailed pattern of the nature and extent of occupational disability among construction workers. METHODS: A cohort study was set up including 14,474 male workers from the construction industry in Wurttemberg (Germany) aged 25-64 years who underwent occupational health exams between 1986 and 1992. The cohort was linked to the regional pension register of the manual workers' pension insurance institution to identify workers who were granted a disability pension during the 10 year follow up. All-cause and cause specific standardised incidence ratios (SIR) and 95% confidence intervals (CI) were calculated using disability rates from the general workforce and from all blue collar workers in Germany as references. RESULTS: In total, 2247 (16%) members of the cohort were granted a disability pension. Major causes of disability were musculoskeletal (45%) and cardiovascular diseases (19%). In comparison with the general workforce, construction workers experienced a higher risk of disability from cancer (SIR = 1.26; 95% CI 1.08 to 1.47), respiratory diseases (SIR = 1.27; 95% CI 1.03 to 1.55), musculoskeletal diseases (SIR = 2.16; 95% CI 2.03 to 2.30), injuries/poisoning (SIR = 2.52; 95% CI 2.06 to 3.05), and all causes combined (SIR = 1.47; 95% CI 1.41 to 1.53). When compared with the blue collar reference group, increased risks of disability among construction workers were found for musculoskeletal diseases (SIR = 1.53; 95% CI 1.44 to 1.63), injury/poisoning (SIR = 1.83; 95% CI 1.50 to 2.21), and all causes combined (SIR = 1.11; 95% CI 1.07 to 1.16). CONCLUSIONS: Musculoskeletal diseases and external causes are major factors limiting the work capability of construction workers and lead to an increased proportion of occupational disability. PMID- 16046610 TI - Osteoarthritis and meniscus disorders of the knee as occupational diseases of miners. AB - AIM: To determine whether kneeling or squatting for prolonged periods is sufficiently causally associated with an increased risk of injury or degenerative disease of the knee joint as to meet the classic criteria to be considered an occupational disease of coal miners for whom these are or have been routine working postures. METHOD: Systematic literature searches were made for studies relating to kneeling and squatting as part of the working environment of coal mines and the role of these postures in causation of knee disorders in coal miners, analogous occupations, populations, and communities. The working environment and potentially damaging forces on the knee when kneeling or squatting were described. Papers on the incidence or prevalence of knee disorders in occupational and other groups were scored against five criteria independently by each author, and from this a single consensus score representing the overall strength of evidence given by the research was awarded. The evidence was then weighed against the criteria for an occupational disease. RESULTS: Nineteen published papers were scored, the majority of which focussed on osteoarthritis as the outcome of interest. Few of the studies found focussed specifically on miners, and those that did tended to involve small numbers of subjects and were carried out before 1960, when the mining population was at its largest but epidemiological evidence of the risk factors for knee disorders was not well established. The non-mining studies in the review represent groups of workers with a similar or lesser kneeling content in their work. CONCLUSION: The papers reviewed provide sufficient evidence to conclude that work involving kneeling and/or squatting is causally associated with an increased risk of osteoarthritis of the knee. In some of the more recent epidemiologically sound studies, frequent or prolonged kneeling or squatting doubles the general risk of osteoarthritis of the knees found in the general population. This may be of particular importance in welfare and medico-legal situations. There was also evidence to suggest that lifting, in combination with kneeling/squatting, an activity also performed by miners in the course of their work, is associated with an excess risk of osteoarthritis above that attributed to kneeling/squatting alone. PMID- 16046611 TI - Short term changes in lung function, leukocytosis in blood, and lachrymal fluid among bacterial single cell protein workers after an episode with high exposure to endotoxins. AB - AIMS: To study possible effects of endotoxin exposure among bacterial single cell protein workers on pulmonary function, blood parameters, and lachrymal fluid before and after a work shift. METHODS: The study included 23 men and five women who were examined at the start and at the end of a work shift. Most workers performed a task with unusually high exposure levels. Twelve of the workers were re-examined the day after. The workers were divided into three exposure groups: production workers with the highest assumed exposure levels (n = 18), engineers (n = 5), and clerks (n = 2). The median endotoxin level during a work shift was 34000 EU/m3 in the high exposure group (range 3300-89000 EU/m3 ), 11000 EU/m3 (range 350-27000 EU/m3) among the engineers, and 180 EU/m3 (range 60-300 EU/m3) for the clerks. The workers answered a questionnaire about work related symptoms. Assessment of lung function included dynamic lung volumes and flows. The blood analysis included cell count of leukocytes and mediators of inflammation, fibrinogen, interleukin-6 (IL-6), D-dimer, and C-reactive protein (CRP). Cells in lachrymal fluid were counted with a microscope. RESULTS: The forced vital capacity (FVC) changed significantly (p<0.05) from 5.34 l (SD 0.9) to 5.25 l (SD 0.9) and forced expired volume in one second (FEV1) from 4.15 l (SD 0.7) to 4.07 l (SD 0.7) during the work shift. The leukocytes increased significantly (p<0.05) from 6.9 10(9)/l (SD 1.2) to 7.7 10(9)/l (SD 1.5) and IL-6 from 1.5 ng/l (SD 0.6) to 3.31 ng/l (SD 2.7). Except for fibrinogen, which had a borderline increase and PEF that decreased, the parameters were normalised the day after. Four of the workers had an increase of neutrofile granulocytes in the lachrymal fluid during the shift. There was a significant association between the endotoxin concentration and decrease of FEV1 despite the use of powered respirators. CONCLUSIONS: During a work shift with unusual high levels of endotoxins at a plant manufacturing bacterial single cell protein the results show that FVC and FEV1 were reduced. Mediators of inflammation increased along with leucocytosis in blood and lachrymal fluid among the workers. PMID- 16046612 TI - World at work: cleaners. PMID- 16046613 TI - Analysis of allosterism in functional assays. AB - The theoretical basis for analyzing the effects of an allosteric modulator on the response to an agonist is described. The effects of an allosteric modulator on the concentration-response curve to an agonist can be attributed to changes in the observed dissociation constant and intrinsic efficacy of the agonist-receptor complex. Each of these two changes can be represented by a coefficient or factor. It is possible to estimate the ratio of the coefficient of change in agonist efficacy divided by that for the agonist dissociation constant. This ratio is designated as the relative activity (RA) of the agonist in the presence of the allosteric modulator. The RA value can be estimated for each concentration of allosteric modulator by nonlinear regression analysis, regardless of the shape of the concentration-response curve. Regression analysis of the RA values against the concentration of allosteric modulator yields estimates of the dissociation constant (K(A)) of the allosteric modulator and the maximal RA value. If the concentration-response curve to the agonist obeys a logistic function and the allosteric modulator influences the maximal response, it is possible to distinguish between the maximal change in affinity from that of efficacy. If the agonist concentration-response curve obeys a logistic equation with a Hill slope of 1, the RA values can be estimated easily from the agonist EC(50) and E(max) values. This analysis illustrates the utility of the RA value in quantifying allosteric effects. PMID- 16046614 TI - Effects of phytoestrogens genistein and daidzein on prostacyclin production by human endothelial cells. AB - The molecular mechanisms of the vascular effects of phytoestrogens are poorly studied. Prostacyclin is a potent vasodilator synthesized by two isoforms of cyclooxygenase (COX) in endothelium. This study examine the effects of two phytoestrogens, the isoflavones genistein and daidzein, on prostacyclin production by cultured human umbilical vein endothelial cells (HUVECs) and the possible role of not only estrogen receptors but also both COX isoforms. The two phytoestrogens significantly increased prostacyclin release in a time- and dose dependent (0.01-1 microM) manner, being higher than control after 24 h. Selective inhibitors of COX-1, SC-560 [5-(4-chlorophenyl)-1-(4-methoxypjenyl)-3 (trifluoromethyl)-1H-pyrazole], and COX-2, NS-398 (N-[2-(cyclohexyloxy)-4 nitrophenyl]-methanesulfonamide), were used to investigate the relative contribution of each enzyme. Both inhibitors decreased basal production of prostacyclin, but only COX-2 inhibition completely abolished the isoflavone stimulated prostacyclin production. Phytoestrogens also increased COX-2 mRNA expression and protein content without affecting COX-1 levels. All these effects were mediated through estrogen receptor activation since treatment of cells with the estrogen receptor antagonist ICI 182780 [7alpha-[9[(4,4,5,5,5 pentafluoropentyl)sulfinyl]nonyl]-estra-1,3,5(10)-triene-3,17beta diol] completely abolished the isoflavone-induced increase in prostacyclin production, COX-2 mRNA expression, and COX-2 protein content. The results clearly support the hypothesis that genistein and daidzein increased HUVEC prostacyclin production through estrogen receptor-dependent mechanism, which involved the enhancement of COX-2 protein and activity. PMID- 16046615 TI - Inhibition of autoantigen expression by (-)-epigallocatechin-3-gallate (the major constituent of green tea) in normal human cells. AB - Autoimmune disorders, characterized by inflammation and apoptosis of target cells leading to tissue destruction, are mediated in part by autoantibodies against normal cellular components (autoantigens) that may be overexpressed. For example, antibodies against the autoantigens SS-A/Ro and SS-B/La are primary markers for systemic lupus erythematosus and Sjogren's syndrome. Recently, studies in animals demonstrated that green tea consumption may reduce the severity of some autoimmune disorders, but the mechanism is unclear. Herein, we sought to determine whether the most abundant green tea polyphenol, (-)-epigallocatechin-3 gallate (EGCG), affects autoantigen expression in human cells. Cultures of pooled normal human primary epidermal keratinocytes and of an immortalized human salivary acinar cell line were incubated with 100 microM EGCG (a physiologically achievable level for topical application or oral administration) for various time periods and then analyzed by cDNA microarray analysis, reverse transcription polymerase chain reaction, and Western blotting for expression of several major autoantigen candidates. EGCG inhibited the transcription and translation of major autoantigens, including SS-B/La, SS-A/Ro, coilin, DNA topoisomerase I, and alpha fodrin. These findings, taken together with green tea's anti-inflammatory and antiapoptotic effects, suggest that green tea polyphenols could serve as an important component in novel approaches to combat autoimmune disorders in humans. PMID- 16046616 TI - The effect of dipyridamole on vascular cell-derived reactive oxygen species. AB - Platelet and vascular stimulation leads to release of reactive oxygen species (ROS) that are known to influence vascular reactivity and thrombosis. Dipyridamole is a vasodilator and platelet inhibitor that has previously been shown to have direct antioxidant properties. The antioxidant effects of dipyridamole on vascular cell-derived ROS are not known; therefore, dipyridamole was incubated with endothelial cells and platelets and cellular redox status and release of endogenous ROS were assessed. Dipyridamole decreased intracellular basal ROS generation from endothelial cells as measured by DCFDA (2',7' dichlorodihydrofluorescein diacetate) oxidation. Incubation of endothelial cells with dipyridamole also attenuated t-butylhydroperoxide-induced oxidative stress. Using a redox-sensitive fluorescent dye, dipyridamole improved cellular activity after treatment with t-butylhydroperoxide. Incubation with dipyridamole did not alter platelet release of nitric oxide or hydrogen peroxide but significantly attenuated superoxide release. Using flow cytometry and confocal microscopy, dipyridamole decreased platelet ROS generation. Dipyridamole also suppressed platelet-soluble CD40 ligand release. In summary, at therapeutically relevant concentrations, dipyridamole suppresses the formation of ROS in platelets and endothelial cells and improves cellular redox status. These data suggest that the redox-dependent properties of dipyridamole have a direct effect on vascular cells. PMID- 16046617 TI - HIF-1alpha-targeted pathways are activated by heat acclimation and contribute to acclimation-ischemic cross-tolerance in the heart. AB - Hypoxia-inducible factor-1 (HIF-1) is a key regulator of the cellular hypoxic response. We previously showed that HIF-1 activation is essential for heat acclimation (AC) in Caenorhabditis elegans. Metabolic changes in AC rat hearts indicate HIF-1alpha activation in mammals as well. Here we characterize the HIF 1alpha profile and the transcriptional activation of its target genes following AC and following heat stress (HS) in hearts from nonacclimated (C; 24 degrees C) and AC (34 degrees C, 1 mo) rats. We used Western blot and immunohistochemistry to measure HIF-1alpha levels and EMSA and RT-PCR/quantitative RT-PCR to detect expression of the HIF-1alpha-targeted genes, including vascular endothelial growth factor (Vegf), heme oxygenase-1 (HO1), erythropoietin (Epo), and Epo receptor (EpoR). EpoR and Epo mRNA levels were measured to determine systemic effects in the kidneys and cross-tolerance effects in C and AC ischemic hearts (Langendorff, 75% ischemia, 40 min). The results demonstrated that 1) after AC, HIF-1alpha protein levels were increased, 2) HS alone induced transient HIF 1alpha upregulation, and 3) VEGF and HO1 mRNA levels increased after HS, with greater magnitude in the AC hearts. Epo mRNA in AC kidneys and EpoR mRNA in AC hearts were also elevated. In AC hearts, EpoR expression was markedly higher after HS or ischemia. Hearts from AC rats were dramatically protected against infarction after ischemia-perfusion. We conclude that HIF-1 contributes to the acclimation-ischemia cross-tolerance mechanism in the heart by induction of both chronic and inducible adaptive components. PMID- 16046619 TI - Genetic and pharmacological inactivation of adenosine A2A receptor reveals an Egr 2-mediated transcriptional regulatory network in the mouse striatum. AB - The adenosine A2A receptor (A2AR) is highly expressed in the striatum, where it modulates motor and emotional behaviors. We used both microarray and bioinformatics analyses to compare gene expression profiles by genetic and pharmacological inactivation of A2AR and inferred an A2AR-controlled transcription network in the mouse striatum. A comparison between vehicle (VEH) treated A2AR knockout (KO) mice (A2AR KO-VEH) and wild-type (WT) mice (WT-VEH) revealed 36 upregulated genes that were partially mimicked by treatment with SCH 58261 (SCH; an A2AR antagonist) and 54 downregulated genes that were not mimicked by SCH treatment. We validated the A2AR as a specific drug target for SCH by comparing A2AR KO-SCH and A2AR KO-VEH groups. The unique downregulation effect of A2AR KO was confirmed by comparing A2AR KO-SCH with WT-SCH gene groups. The distinct striatal gene expression profiles induced by A2AR KO and SCH should provide clues to the molecular mechanisms underlying the different phenotypes observed after genetic and pharmacological inactivation of A2AR. Furthermore, bioinformatics analysis discovered that Egr-2 binding sites were statistically overrepresented in the proximal promoters of A2AR KO-affected genes relative to the unaffected genes. This finding was further substantiated by the demonstration that the Egr-2 mRNA level increased in the striatum of both A2AR KO and SCH treated mice and that striatal Egr-2 binding activity in the promoters of two A2AR KO-affected genes was enhanced in A2AR KO mice as assayed by chromatin immunoprecipitation. Taken together, these results strongly support the existence of an Egr-2-directed transcriptional regulatory network controlled by striatal A2ARs. PMID- 16046620 TI - Alterations of nuclear envelope and chromatin organization in mandibuloacral dysplasia, a rare form of laminopathy. AB - Autosomal recessive mandibuloacral dysplasia [mandibuloacral dysplasia type A (MADA); Online Mendelian Inheritance in Man (OMIM) no. 248370] is caused by a mutation in LMNA encoding lamin A/C. Here we show that this mutation causes accumulation of the lamin A precursor protein, a marked alteration of the nuclear architecture and, hence, chromatin disorganization. Heterochromatin domains are altered or completely lost in MADA nuclei, consistent with the finding that heterochromatin-associated protein HP1beta and histone H3 methylated at lysine 9 and their nuclear envelope partner protein lamin B receptor (LBR) are delocalized and solubilized. Both accumulation of lamin A precursor and chromatin defects become more severe in older patients. These results strongly suggest that altered chromatin remodeling is a key event in the cascade of epigenetic events causing MADA and could be related to the premature-aging phenotype. PMID- 16046621 TI - The implications of higher (or lower) success in secondary structure prediction of chain fragments. PMID- 16046618 TI - Gene expression profiling of gilthead sea bream during early development and detection of stress-related genes by the application of cDNA microarray technology. AB - Large-scale gene expression studies were performed for one of the main European aquaculture species, the gilthead sea bream Sparus auratus L. For this purpose, a cDNA microarray containing 10,176 clones from a cDNA library of mixed embryonic and larval stages was constructed. In addition to its importance for aquaculture, the taxonomic position and the relatively small genome size of sea bream makes it a prospective model for evolutionary biology and comparative genomics. However, so far, no large-scale analysis of gene expression exists for this species. In the present study, gene expression was analyzed in gilthead sea bream during early development, a significant period in the determination of quantitative traits and therefore of considerable interest for aquaculture. Synexpression groups expressed primarily early and late in development were determined and were composed of both known and novel genes. Furthermore, it was possible to identify stress response genes induced by cortisol injections using the cDNA microarray generated. The creation of gene expression profiles for sea bream by microarray hybridization will accelerate identification of candidate genes involved in multifactorial traits and certain regulatory pathways and will also contribute to a better understanding of the genetic background of fish physiology, which may help to improve aquaculture practices. PMID- 16046622 TI - Remodeling protein complexes: insights from the AAA+ unfoldase ClpX and Mu transposase. AB - Multiprotein complexes in the cell are dynamic entities that are constantly undergoing changes in subunit composition and conformation to carry out their functions. The protein-DNA complex that promotes recombination of the bacteriophage Mu is a prime example of a complex that must undergo specific changes to carry out its function. The Clp/Hsp100 family of AAA+ ATPases plays a critical role in mediating such changes. The Clp/Hsp100 unfolding enzymes have been extensively studied for the roles they play in protein degradation. However, degradation is not the only fate for proteins that come in contact with the ATP dependent unfolding enzymes. The Clp/Hsp100 enzymes induce structural changes in their substrates. These structural changes, which we refer to as "remodeling", ultimately change the biological activity of the substrate. These biological changes include activation, inactivation (not associated with degradation), and relocation within the cell. Analysis of the interaction between Escherichia coli ClpX unfoldase and the Mu recombination complex, has provided molecular insight into the mechanisms of protein remodeling. We discuss the key mechanistic features of the remodeling reactions promoted by ClpX and possible implications of these findings for other biological reactions. PMID- 16046623 TI - Crystal structure of the human vascular adhesion protein-1: unique structural features with functional implications. AB - The expression of human vascular adhesion protein-1 (hVAP-1) is induced at sites of inflammation where extravasation of lymphocytes from blood to the peripheral tissue occurs. We have solved the X-ray structure of hVAP-1, a human copper amine oxidase (CAO), which is distinguished from other CAOs in being membrane-bound. The dimer structure reveals some intriguing features that may have fundamental roles in the adhesive and enzymatic functions of hVAP-1, especially regarding the role of hVAP-1 in inflammation, lymphocyte attachment, and signaling. Firstly, Leu469 at the substrate channel may play a key role in controlling the substrate entry; depending on its conformation, it either blocks or gives access to the active site. Secondly, sugar units are clearly observed at two of the six predicted N-glycosylation sites. Moreover, mutagenesis analysis showed that all of the predicted sites were glycosylated in the protein used for crystallization. Thirdly, the existence of a solvent-exposed RGD motif at the entrance to each active site in hVAP-1 suggests that it may have a functional role. PMID- 16046624 TI - The intrinsically disordered C-terminal domain of the measles virus nucleoprotein interacts with the C-terminal domain of the phosphoprotein via two distinct sites and remains predominantly unfolded. AB - Measles virus is a negative-sense, single-stranded RNA virus within the Mononegavirales order,which includes several human pathogens, including rabies, Ebola, Nipah, and Hendra viruses. The measles virus nucleoprotein consists of a structured N-terminal domain, and of an intrinsically disordered C-terminal domain, N(TAIL) (aa 401-525), which undergoes induced folding in the presence of the C-terminal domain (XD, aa 459-507) of the viral phosphoprotein. With in N(TAIL), an alpha-helical molecular recognition element (alpha-MoRE, aa 488-499) involved in binding to P and in induced folding was identified and then observed in the crystal structure of XD. Using small-angle X-ray scattering, we have derived a low-resolution structural model of the complex between XD and N(TAIL), which shows that most of N(TAIL) remains disordered in the complex despite P induced folding within the alpha-MoRE. The model consists of an extended shape accommodating the multiple conformations adopted by the disordered N-terminal region of N(TAIL), and of a bulky globular region, corresponding to XD and to the C terminus of N(TAIL) (aa 486-525). Using surface plasmon resonance, circular dichroism, fluorescence spectroscopy, and heteronuclear magnetic resonance, we show that N(TAIL) has an additional site (aa 517-525) involved in binding to XD but not in the unstructured-to-structured transition. This work provides evidence that intrinsically disordered domains can establish complex interactions with their partners, and can contact them through multiple sites that do not all necessarily gain regular secondary structure. PMID- 16046625 TI - Crystal structure of Yersinia enterocolitica type III secretion chaperone SycT. AB - Pathogenic Yersinia species use a type III secretion (TTS) system to deliver a number of cytotoxic effector proteins directly into the mammalian host cell. To ensure effective translocation, several such effector proteins transiently bind to specific chaperones in the bacterial cytoplasm. Correspondingly, SycT is the chaperone of YopT, a cysteine protease that cleaves the membrane-anchor of Rho GTPases in the host. We have analyzed the complex between YopT and SycT and determined the structure of SycT in three crystal forms. Biochemical studies indicate a stoichometric effector/chaperone ratio of 1:2 and the chaperone binding site contains at least residues 52-103 of YopT. The crystal structures reveal a SycT homodimer with an overall fold similar to that of other TTS effector chaperones. In contrast to the canonical five-stranded anti-parallel beta-sheet flanked by three alpha-helices, SycT lacks the dimerization alpha helix and has an additional beta-strand capable of undergoing a conformational change. The dimer interface consists of two beta-strands and the connecting loops. Two hydrophobic patches involved in effector binding in other TTS effector chaperones are also found in SycT. The structural similarity of SycT to other chaperones and the spatial conservation of effector-binding sites support the idea that TTS effector chaperones form a single functional and structural group. PMID- 16046626 TI - Interdomain side-chain interactions in human gammaD crystallin influencing folding and stability. AB - Human gammaD crystallin (HgammaD-Crys) is a two domain, beta-sheet eye lens protein that must remain soluble throughout life for lens transparency. Single amino acid substitutions of HgammaD-Crys are associated with juvenile-onset cataracts. Features of the interface between the two domains conserved among gamma-crystallins are a central six-residue hydrophobic cluster, and two pairs of interacting residues flanking the cluster. In HgammaD-Crys these pairs are Gln54/Gln143 and Arg79/Met147. We previously reported contributions of the hydrophobic cluster residues to protein stability. In this study alanine substitutions of the flanking residue pairs were constructed and analyzed. Equilibrium unfolding/refolding experiments at 37 degrees C revealed a plateau in the unfolding/refolding transitions, suggesting population of a partially folded intermediate with a folded C-terminal domain (C-td) and unfolded N-terminal domain (N-td). The N-td was destabilized by substituting residues from both domains. In contrast, the C-td was not significantly affected by substitutions of either domain. Refolding rates of the N-td were significantly decreased for mutants of either domain. In contrast, refolding rates of the C-td were similar to wild type for mutants of either domain. Therefore, domain interface residues of the folded C-td probably nucleate refolding of the N-td. We suggest that these residues stabilize the native state by shielding the central hydrophobic cluster from solvent. Glutamine and methionine side chains are among the residues covalently damaged in aged and cataractous lenses. Such damage may generate partially unfolded, aggregation- prone conformations of HgammaD-Crys that could be significant in cataract. PMID- 16046627 TI - Soluble domains of telomerase reverse transcriptase identified by high-throughput screening. AB - Telomerase is a ribonucleoprotein complex responsible for extending the ends of eukaryotic chromosomes. Structural and biophysical studies of this enzyme have been limited by the inability to produce large amounts of recombinant protein. Here we perform a high-throughput screen to map regions of the Tetrahymena thermophila TERT (Telomerase Reverse Transcriptase) protein that are overexpressed in a soluble form in Escherichia coli using a GFP-fusion system. Many of the soluble protein domains identified do not coincide with domains inferred from multiple sequence alignment, so screening for fluorescent colonies provided information not otherwise readily obtained. The method revealed an essential, independently folded N-terminal domain that was expressed and purified with high yield and found to be suitable for structural analysis. These results provide a tool for future structural and biophysical studies of TERT. PMID- 16046628 TI - Design of enhanced agonists through the use of a new virtual screening method: application to peptides that bind class I major histocompatibility complex (MHC) molecules. AB - A new screening procedure is described that uses docking calculations to design enhanced agonist peptides that bind to major histocompatibility complex (MHC) class I receptors. The screening process proceeds via single mutations of one amino acid at the positions that directly interact with the MHC receptor. The energetic and structural effects of these mutations have been studied using fragments of the original ligand that vary in length. The results of these docking studies indicate that the mutant affinity ranking of long peptides can be practically reproduced with a screening approach performed using fragments of six residues. Fragments of four and five residues could mimic, in some cases, the structural arrangement of the side chains of the full-length peptide. We have compared the structural and energetic results of the docking calculations with experimental data using three unrelated ligand peptides that differ greatly in their affinity for the MHC complex. Analysis of the affinity of the fragments led to the identification of three important parameters in the construction of fragments that mimic the structural and energetic properties of the full-length ligand: the length of the fragment; its intermolecular energy; and the number and localization, internal or terminal, of the anchor residues. The results of this new peptide-design methodology have been applied to suggest new peptides derived from the MUC1-8 peptide that could be used as murine vaccines that trigger the immune response through the MHC class I protein H-2K(b). PMID- 16046629 TI - Three-dimensional structure of the AAH26994.1 protein from Mus musculus, a putative eukaryotic Urm1. AB - We have used NMR spectroscopy to determine the solution structure of protein AAH26994.1 from Mus musculus and propose that it represents the first three dimensional structure of a ubiquitin-related modifier 1 (Urm1) protein. Amino acid sequence comparisons indicate that AAH26994.1 belongs to the Urm1 family of ubiquitin-like modifier proteins. The best characterized member of this family has been shown to be involved in nutrient sensing, invasive growth, and budding in yeast. Proteins in this family have only a weak sequence similarity to ubiquitin, and the structure of AAH26994.1 showed a much closer resemblance to MoaD subunits of molybdopterin synthases (known structures are of three bacterial MoaD proteins with 14%-26% sequence identity to AAH26994.1). The structures of AAH26994.1 and the MoaD proteins each contain the signature ubiquitin secondary structure fold, but all differ from ubiquitin largely in regions outside of this fold. This structural similarity bolsters the hypothesis that ubiquitin and ubiquitin-related proteins evolved from a protein-based sulfide donor system of the molybdopterin synthase type. PMID- 16046630 TI - A novel system for continuous protein refolding and on-line capture by expanded bed adsorption. AB - A novel two-step protein refolding strategy has been developed, where continuous renaturation-bydilution is followed by direct capture on an expanded bed adsorption (EBA) column. The performance of the overall process was tested on a N terminally tagged version of human beta2-microglobulin (HAT-hbeta2m) both at analytical, small, and preparative scale. In a single scalable operation, extracted and denatured inclusion body proteins from Escherichia coli were continuously diluted into refolding buffer, using a short pipe reactor, allowing for a defined retention and refolding time, and then fed directly to an EBA column, where the protein was captured, washed, and finally eluted as soluble folded protein. Not only was the eluted protein in a correctly folded state, the purity of the HAThbeta2m was increased from 34% to 94%, and the product was concentrated sevenfold. The yield of the overall process was 45%, and the product loss was primarily a consequence of the refolding reaction rather than the EBA step. Full biological activity of HAT-hbeta2m was demonstrated after removal of the HAT-tag. In contrast to batch refolding, a continuous refolding strategy allows the conditions to be controlled and maintained throughout the process, irrespective of the batch size; i.e., it is readily scalable. Furthermore, the procedure is fast and tolerant toward aggregate formation, a common complication of in vitro protein refolding. In conclusion, this system represents a novel approach to small and preparative scale protein refolding, which should be applicable to many other proteins. PMID- 16046631 TI - Influence of the N-terminal domain on the aggregation properties of the prion protein. AB - Prion diseases appear to be caused by the aggregation of the cellular prion protein (PrP(C)) into an infectious form denoted PrP(Sc). The in vitro aggregation of the prion protein has been extensively investigated, yet many of these studies utilize truncated polypeptides. Because the C-terminal portion of PrP(Sc) is protease-resistant and retains infectivity, it is assumed that studies on this fragment are most relevant. The full-length protein can be distinguished from the truncated protein because it contains a largely structured, alpha helical, C-terminal region in addition to an N terminus that is unstructured in the absence of metal ion binding. Herein, the in vitro aggregation of a truncated portion of the prion protein (PrP 90-231) and a full-length version (PrP 23-231) were compared. In each case, concentration-dependent aggregation was analyzed to discern whether it proceeds by a nucleation-dependent pathway. Both protein constructs appear to aggregate via a nucleated polymerization with a small nucleus size, yet the later steps differ. The full-length protein forms larger aggregates than the truncated protein, indicating that the N terminus may mediate higher-order aggregation processes. In addition, the N terminus has an influence on the assembly state of PrP before aggregation begins, causing the full-length protein to adopt several oligomeric forms in a neutral pH buffer. Our results emphasize the importance of studying the full-length protein in addition to the truncated forms for in vitro aggregation studies in order to make valid hypotheses about the mechanisms of prion aggregation and the distribution of aggregates in vivo. PMID- 16046632 TI - HCA and HML isolated from the red marine algae Hypnea cervicornis and Hypnea musciformis define a novel lectin family. AB - HCA and HML represent lectins isolated from the red marine algae Hypnea cervicornis and Hypnea musciformis, respectively. Hemagglutination inhibition assays suggest that HML binds GalNAc/Gal substituted with a neutral sugar through 1-3, 1-4, or 1-2 linkages in O-linked mucin-type glycans, and Fuc(alpha1-6)GlcNAc of N-linked glycoproteins. The specificity of HCA includes the epitopes recognized by HML, although the glycoproteins inhibited distinctly HML and HCA. The agglutinating activity of HCA was inhibited by GalNAc, highlighting the different fine sugar epitope-recognizing specificity of each algal lectin. The primary structures of HCA (9193+/-3 Da) and HML (9357+/-1 Da) were determined by Edman degradation and tandem mass spectrometry of the N-terminally blocked fragments. Both lectins consist of a mixture of a 90-residue polypeptide containing seven intrachain disulfide bonds and two disulfide-bonded subunits generated by cleavage at the bond T50-E51 (HCA) and R50-E51 (HML). The amino acid sequences of HCA and HML display 55% sequence identity (80% similarity) between themselves, but do not show discernible sequence and cysteine spacing pattern similarities with any other known protein structure, indicating that HCA and HML belong to a novel lectin family. Alignment of the amino acid sequence of the two lectins revealed the existence of internal domain duplication, with residues 1-47 and 48-90 corresponding to the N- and C-terminal domains, respectively. The six conserved cysteines in each domain may form three intrachain cysteine linkages, and the unique cysteine residues of the N-terminal (Cys46) and the C-terminal (Cys71) domains may form an intersubunit disulfide bond. PMID- 16046633 TI - Refractive index-based determination of detergent concentration and its application to the study of membrane proteins. AB - The concentration of detergent in membrane protein preparations can have a critical role on protein stability, function, and the potential for crystallization. Unfortunately, dialysis or protein concentration can lead to an unknown amount of detergent in the final membrane protein preparations. Here we present a method for the determination of detergent concentration based on refractive index of the detergent solution. This method was applied to quantitate the amount of detergent remaining in solution after concentration in various concentrators. We found that the ability of the tested detergents to pass through the molecular weight cutoff membrane correlates well with detergent micelle size. Therefore, the micelle size can be used as a rough guide to estimate the retention of a given detergent in various molecular weight cutoff concentrators. The refractive index method is exceptionally informative when coupled with size exclusion chromatography and light scattering, and can be used to determine the oligomeric state of the membrane protein, the size of a protein-associated micelle, as well as the amount and size of the unbound detergent micelle. PMID- 16046634 TI - The cover. The Happy Moment. PMID- 16046635 TI - MicroRNAs move into cancer research. PMID- 16046636 TI - Medical groups sue EPA over mercury rule. PMID- 16046637 TI - AMA, AAMC say reform needed across continuum of US medical education. PMID- 16046644 TI - Vitamin E supplementation, cardiovascular events, and cancer. PMID- 16046645 TI - Vitamin E supplementation, cardiovascular events, and cancer. PMID- 16046646 TI - Vitamin E supplementation, cardiovascular events, and cancer. PMID- 16046647 TI - Physician substance abuse. PMID- 16046648 TI - Comparison of drug regimen costs between the Medicare prescription discount program and other purchasing systems. PMID- 16046649 TI - Risk of prostate cancer-specific mortality following biochemical recurrence after radical prostatectomy. AB - CONTEXT: The natural history of biochemical recurrence after radical prostatectomy can be long but variable. Better risk assessment models are needed to identify men who are at high risk for prostate cancer death early and who may benefit from aggressive salvage treatment and to identify men who are at low risk for prostate cancer death and can be safely observed. OBJECTIVES: To define risk factors for prostate cancer death following radical prostatectomy and to develop tables to risk stratify for prostate cancer-specific survival. DESIGN, SETTING, AND PATIENTS: Retrospective cohort study of 379 men who had undergone radical prostatectomy at an urban tertiary care hospital between 1982 and 2000 and who had a biochemical recurrence and after biochemical failure had at least 2 prostate-specific antigen (PSA) values at least 3 months apart in order to calculate PSA doubling time (PSADT). The mean (SD) follow-up after surgery was 10.3 (4.7) years and median follow-up was 10 years (range, 1-20 years). MAIN OUTCOME MEASURE: Prostate cancer-specific mortality. RESULTS: Median survival had not been reached after 16 years of follow-up after biochemical recurrence. Prostate-specific doubling time (<3.0 vs 3.0-8.9 vs 9.0-14.9 vs > or =15.0 months), pathological Gleason score (< or =7 vs 8-10), and time from surgery to biochemical recurrence (< or =3 vs >3 years) were all significant risk factors for time to prostate-specific mortality. Using these 3 variables, tables were constructed to estimate the risk of prostate cancer-specific survival at year 15 after biochemical recurrence. CONCLUSION: Clinical parameters (PSADT, pathological Gleason score, and time from surgery to biochemical recurrence) can help risk stratify patients for prostate cancer-specific mortality following biochemical recurrence after radical prostatectomy. These preliminary findings may serve as useful guides to patients and their physicians to identify patients at high risk for prostate cancer-specific mortality following biochemical recurrence after radical prostatectomy to enroll them in early aggressive treatment trials. In addition, these preliminary findings highlight that survival in low-risk patients can be quite prolonged. PMID- 16046650 TI - Pretreatment PSA velocity and risk of death from prostate cancer following external beam radiation therapy. AB - CONTEXT: Men with localized prostate cancer and a preoperative prostate-specific antigen (PSA) velocity greater than 2.0 ng/mL per year experience a 10-fold increase in prostate cancer-specific mortality despite surgery. OBJECTIVE: To assess whether a greater than 2.0-ng/mL increase in PSA level during the year prior to diagnosis was significantly associated with prostate cancer-specific mortality following radiation therapy (RT). DESIGN, SETTING, AND PATIENTS: Between January 1, 1989, and December 1, 2002, 358 men treated with RT for localized prostate cancer formed the study cohort (median age at treatment, 71.2 [range, 43.2-83.5] years). A Cox regression multivariable analysis was used to evaluate whether a PSA velocity greater than 2.0 ng/mL per year was significantly associated with prostate cancer-specific mortality and all-cause mortality after controlling for prognostic factors available at diagnosis. MAIN OUTCOME MEASURE: Time to prostate cancer-specific mortality for the 125 men with low-risk prostate cancer (clinical tumor category T1c or T2a and PSA level <10.0 ng/mL and Gleason score < or =6) and the 233 men with higher-risk disease, stratified by the PSA velocity. RESULTS: A PSA velocity greater than 2.0 ng/mL per year was significantly associated with a shorter time to prostate cancer-specific mortality (adjusted hazard ratio [HR], 12.0; 95% confidence interval [CI], 3.0 54.0; P = .001) and all-cause mortality (adjusted HR, 2.1; 95% CI, 1.3-3.6; P = .005) when compared with men whose PSA velocity was 2.0 ng/mL per year or less. Men presenting with low-risk disease and a PSA velocity greater than 2.0 ng/mL per year had a 7-year estimate of prostate cancer-specific mortality of 19% (95% CI, 2%-39%) compared with 0% for men whose PSA velocity was 2.0 ng/mL per year or less. The corresponding values for men with higher-risk disease were 24% (95% CI, 12%-37%) and 4% (95% CI, 0%-11%), respectively. CONCLUSIONS: A greater than 2.0 ng/mL increase in PSA level during the year prior to diagnosis is associated with a significantly higher risk of death due to prostate cancer following RT despite having low-risk disease. Such men who are planning to undergo RT and are in good health could be considered for RT combined with androgen suppression therapy because this approach improves survival in men with higher-risk disease. PMID- 16046651 TI - Trends in management and outcomes of patients with acute myocardial infarction complicated by cardiogenic shock. AB - CONTEXT: Early mechanical revascularization in patients with acute myocardial infarction (AMI) complicated by cardiogenic shock is a therapeutic strategy that reduces mortality. It has been a class I recommendation in guidelines from the American College of Cardiology and the American Heart Association since 1999 for patients younger than 75 years. However, little is known about implementation of these guidelines in practice. OBJECTIVES: To assess trends in early revascularization and mortality for patients with cardiogenic shock complicating AMI and to determine whether the national guidelines affect revascularization rates. DESIGN, SETTING, AND PATIENTS: Prospective, observational study of 293,633 patients with ST-elevation myocardial infarction (25,311 [8.6%] had cardiogenic shock; 7356 [29%] had cardiogenic shock at hospital presentation) enrolled in the National Registry of Myocardial Infarction (NRMI) from January 1995 to May 2004 at 775 US hospitals with revascularization capability (defined as the capability to perform cardiac catheterization, percutaneous coronary intervention [PCI], and open-heart surgery). MAIN OUTCOME MEASURES: Management patterns and in-hospital mortality rates. RESULTS: There was an increase in primary PCI rates from 27.4% to 54.4% (P<.001) in hospitals with revascularization capability that paralleled the change in PCI for ST-elevation myocardial infarction. There was no significant change in rates of immediate coronary artery bypass graft surgery (from 2.1% to 3.2%). Propensity-adjusted multivariable analyses demonstrated that primary PCI was associated with a decreased odds of death during hospitalization (odds ratio, 0.46; 95% confidence interval, 0.40-0.53). There were no differences in the rates of change in revascularization rates based on the date when the guidelines were released regardless of patient age. Overall in-hospital cardiogenic shock mortality decreased from 60.3% in 1995 to 47.9% in 2004 (P<.001). CONCLUSIONS: The use of PCI for patients with cardiogenic shock was associated with improved survival in a large group of hospitals with revascularization capability. The American College of Cardiology and American Heart Association guidelines had no detectable temporal impact on revascularization rates. These findings support the need for increased adherence to these guidelines. PMID- 16046652 TI - Acute illnesses associated with pesticide exposure at schools. AB - CONTEXT: Pesticides continue to be used on school property, and some schools are at risk of pesticide drift exposure from neighboring farms, which leads to pesticide exposure among students and school employees. However, information on the magnitude of illnesses and risk factors associated with these pesticide exposures is not available. OBJECTIVE: To estimate the magnitude of and associated risk factors for pesticide-related illnesses at schools. DESIGN, SETTING, AND PARTICIPANTS: Analysis of surveillance data from 1998 to 2002 of 2593 persons with acute pesticide-related illnesses associated with exposure at schools. Nationwide information on pesticide-related illnesses is routinely collected by 3 national pesticide surveillance systems: the National Institute for Occupational Safety and Health's Sentinel Event Notification System for Occupational Risks pesticides program, the California Department of Pesticide Regulation, and the Toxic Exposure Surveillance System. MAIN OUTCOME MEASURES: Incidence rates and severity of acute pesticide-related illnesses. RESULTS: Incidence rates for 1998-2002 were 7.4 cases per million children and 27.3 cases per million school employee full-time equivalents. The incidence rates among children increased significantly from 1998 to 2002. Illness of high severity was found in 3 cases (0.1%), moderate severity in 275 cases (11%), and low severity in 2315 cases (89%). Most illnesses were associated with insecticides (n = 895, 35%), disinfectants (n = 830, 32%), repellents (n = 335, 13%), or herbicides (n = 279, 11%). Among 406 cases with detailed information on the source of pesticide exposure, 281 (69%) were associated with pesticides used at schools and 125 (31%) were associated with pesticide drift exposure from farmland. CONCLUSIONS: Pesticide exposure at schools produces acute illnesses among school employees and students. To prevent pesticide-related illnesses at schools, implementation of integrated pest management programs in schools, practices to reduce pesticide drift, and adoption of pesticide spray buffer zones around schools are recommended. PMID- 16046653 TI - Hypertension in adults across the age spectrum: current outcomes and control in the community. AB - CONTEXT: Data are sparse regarding current rates of hypertension treatment and control, and risks associated with hypertension, among persons older than 80 years. OBJECTIVE: To determine the prevalence of blood pressure stages, hypertension treatment and control, and cardiovascular risk among older patients with hypertension. DESIGN, SETTING, AND PARTICIPANTS: A community-based cohort study in which data were collected during all Framingham Heart Study examinations attended in the 1990s. Participants were pooled according to age: younger than 60 years, 60 to 79 years, or 80 years or older. There were 5296 participants who contributed 14 458 person-examinations of observation, including 7135 hypertensive person-examinations (4919 treated). MAIN OUTCOME MEASURES: Prevalence of hypertension, its treatment, and its control were compared across age groups. Risks for incident cardiovascular disease during follow-up of up to 6 years were estimated as multivariate-adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) using Cox proportional hazards regression. RESULTS: Prevalence of hypertension and drug treatment increased with advancing age, whereas control rates were markedly lower in older women (systolic <140 and diastolic <90 mm Hg). For ages younger than 60 years, 60 to 79, and 80 years and older, respectively, control rates were 38%, 36%, and 38% in men (P = .30) and 38%, 28%, and 23% in women (P<.001). Relative risks for cardiovascular disease associated with increasing blood pressure stage did not decline with advancing age, and absolute risks increased markedly. Among participants 80 years of age or older, major cardiovascular events occurred in 9.5% of the normal blood pressure (referent) group, 19.8% of the prehypertension group (HR, 1.9; 95% CI, 0.9-3.9), 20.3% of the stage 1 hypertension group (HR, 1.8; 95% CI, 0.8-3.7), and 24.7% of the stage 2 or treated hypertension group (HR, 2.4; 95% CI, 1.2-4.6). CONCLUSIONS: Relative to current national guidelines, rates of blood pressure control in the community are low, especially among older women with hypertension. Short-term risks for cardiovascular disease are substantial, indicating the need for greater efforts at safe, effective risk reduction among the oldest patients with hypertension. PMID- 16046654 TI - Delivery of preventive services to older adults by primary care physicians. AB - CONTEXT: Rates of preventive services remain below national goals. OBJECTIVE: To identify characteristics of physicians and their practices that are associated with the quality of preventive care their patients receive. DESIGN: Cross sectional analysis of data on US physician respondents to the 2000-2001 Community Tracking Study Physician Survey linked to claims data on Medicare beneficiaries they treated in 2001. Physician variables included training and qualifications and sex. Practice setting variables included practice type, size, sources of revenue, and access to information technology. Analyses were adjusted for patient demographics and comorbidity, as well as community characteristics. SETTING AND PARTICIPANTS: Primary care delivered by 3660 physicians providing usual care to 24 581 Medicare beneficiaries aged 65 years and older. MAIN OUTCOME MEASURES: Proportion of eligible beneficiaries receiving each of 6 preventive services: diabetic monitoring with hemoglobin A(1c) measurement or eye examinations, screening for colon or breast cancer, and vaccination for influenza or pneumococcus in 2001. RESULTS: Overall, the proportion of beneficiaries receiving services was below national goals. Physician and, more consistently, practice level characteristics were both associated with differences in the delivery of services. The strongest associations were with practice type and the percentage of practice revenue derived from Medicaid. For instance, beneficiaries receiving usual care in practices with less than 6% of revenue from Medicaid were more likely than those with more than 15% of revenue derived from Medicaid to receive diabetic eye examinations (48.9% vs 43%; P = .02), hemoglobin A1c monitoring (61.2% vs 48.4%; P<.001), mammograms (52.1% vs 38.9%; P<.001), colon cancer screening (10.0% vs 8.5%; P = .60), and influenza (50.2% vs 39.2%; P<.001) and pneumococcal (8.2% vs 6.4%; P<.001) vaccinations. Other variables associated with delivery of preventive services after adjustment for patient and geographic factors included obtaining usual health care from a physician who worked in group practices of 3 or more, who was a graduate of a US or Canadian medical school, or who reported availability of information technology to generate preventive care reminders or access treatment guidelines. CONCLUSIONS: Delivery of routine preventive services is suboptimal for Medicare beneficiaries. However, patients treated within particular practice settings and by particular subgroups of physicians are at particular risk of low-quality care. Profiling these practices may help develop tailored interventions that can be directed to sites where the opportunities for quality improvement are greatest. PMID- 16046655 TI - What to do with a patient who smokes. AB - Despite the reality that smoking remains the most important preventable cause of death and disability, most clinicians underperform in helping smokers quit. Of the 46 million current smokers in the United States, 70% say they would like to quit, but only a small fraction are able to do so on their own because nicotine is so highly addictive. One third to one half of all smokers die prematurely. Reasons clinicians avoid helping smokers quit include time constraints, lack of expertise, lack of financial incentives, respect for a smoker's privacy, fear that a negative message might lose customers, pessimism because most smokers are unable to quit, stigma, and clinicians being smokers. The gold standard for cessation treatment is the 5 As (ask, advise, assess, assist, and arrange). Yet, only a minority of physicians know about these, and fewer put them to use. Acceptable shortcuts are asking, advising, and referring to a telephone "quit line" or an internal referral system. Successful treatment combines counseling with pharmacotherapy (nicotine replacement therapy with or without psychotropic medication such as bupropion). Nicotine replacement therapy comes in long-acting (patch) or short-acting (gum, lozenge, nasal spray, or inhaler) forms. Ways to counter clinicians' pessimism about cessation include the knowledge that most smokers require multiple quit attempts before they succeed, that rigorous studies show long-term quit rates of 14% to 20%, with 1 report as high as 35%, that cessation rates for users of telephone quit lines and integrated health care systems are comparable with those of individual clinicians, and that no other clinical intervention can offer such a large potential benefit. PMID- 16046656 TI - Efficiency in the health care industries: a view from the outside. PMID- 16046657 TI - PSA kinetics and risk of death from prostate cancer: in search of the Holy Grail of surrogate end points. PMID- 16046658 TI - JAMA patient page. Viral encephalitis. PMID- 16046659 TI - Nerve growth factor-independent neuronal survival: a role for NO donors. AB - Because of the limited therapeutic applications of nerve growth factor (NGF), there has been increasing focus on the development of pharmacological tools to bypass the requirement of NGF for the activation of the TrkA tyrosine kinase receptor neuronal survival pathway. In this issue of Molecular Pharmacology, the work by Culmsee et al. (p. 1006) shows that NGF-independent activation of TrkA by protein tyrosine phosphatase (PTP) inhibitors is only achieved when accompanied by release of nitric oxide (NO). This work identifies the integration of the NO/cGMP/protein kinase G (PKG) and NGF/TrkA pathways to induce activation of Akt and ERK1/2 and mediate neuronal survival in the absence of NGF. In addition, it underscores the potential therapeutic effects of ethyl-3,4-dephostatin (DPN), a stable analog of the naturally occurring PTP inhibitor dephostatin, which serves as a NO donor and protects neurons from apoptosis. This Perspective comparatively reviews two major signal transduction pathways that mediate NGF-independent neuronal survival by activating the TrkA pathway: the NO/cGMP/PKG and adenosine/G protein-coupled receptor (GPCR) pathways. PMID- 16046660 TI - Geranylgeranylacetone protects membranes against nonsteroidal anti-inflammatory drugs. AB - Direct gastric mucosal cell damage mediated by nonsteroidal anti-inflammatory drugs (NSAIDs) is involved in the formation of NSAID-induced gastric lesions. We recently suggested that this direct cytotoxicity of NSAIDs is caused by their membrane-permeabilization activity. Geranylgeranylacetone (GGA), a clinically used antiulcer drug, can protect gastric mucosa against lesion formation mediated by NSAIDs. However, the mechanism by which this occurs is not fully understood. In this study, we show that GGA acts to stabilize membranes against NSAIDs. GGA suppressed NSAID-induced permeabilization of calcein-loaded liposomes and NSAID induced stimulation of K(+)-efflux across the cytoplasmic membrane in cells. GGA was effective even when coadministered with NSAIDs and was also able to restore membrane fluidity that had been compromised by NSAIDs. This mechanism seems to play an important role in the antiulcer activity of GGA. PMID- 16046661 TI - Human hepatobiliary transport of organic anions analyzed by quadruple-transfected cells. AB - Hepatobiliary elimination of many organic anions is initiated by OATP1B1 (OATP2, LST-1, OATP-C), OATP1B3 (OATP8), and OATP2B1 (OATP-B), which are the predominant uptake transporters of human hepatocytes. Thereafter, the unidirectional efflux pump ABCC2 (multidrug resistance protein 2) mediates the transport of organic anions, including glutathione conjugates and glucuronosides, into bile. In this study, we generated a Madin-Darby canine kidney (MDCKII) cell line stably expressing recombinant OATP1B1, OATP1B3, and OATP2B1 in the basolateral membrane and ABCC2 in the apical membrane. Double-transfected MDCKII cells stably expressing ABCC2 together with OATP1B1, OATP1B3, or OATP2B1 served as control cells. The quadruple-transfected cells exhibited high rates of vectorial transport of organic anions, including bromosulfophthalein, cholecystokinin peptide (CCK-8), and estrone 3-sulfate. The quadruple-transfected cells enabled the identification of substrates for uptake or vectorial transport that may be missed in studies with a double-transfected cell line, as exemplified by CCK-8, which is a substrate for OATP1B3 but not for OATP1B1 or OATP2B1. The broad substrate spectrum covered by the three hepatocellular OATP transporters enables representative analyses of the uptake of many organic anions into human hepatocytes. The broad spectrum of organic anions transported vectorially by the quadruple-transfected cells also provides valuable information on the substrate selectivity of ABCC2, without the need for studies in inside-out membrane vesicles containing the ABCC2 protein. The quadruple-transfected MDCKII ABCC2/OATP1B1/1B3/2B1 cells may thus be useful for the identification of substrates and inhibitors, including drug candidates, undergoing uptake and secretion by human hepatocytes, under conditions that may be better defined than in primary cultures of human hepatocytes. PMID- 16046662 TI - An investigation of tic suppression and the rebound effect in Tourette's disorder. AB - Many patients, parents of children with Tourette's disorder, and professionals have suggested that following a period of suppression, tics will rebound to a rate that will exceed the average rate of occurrence. At present, there are no empirical data to support or refute such an effect. This experiment utilized an A B-A design with replication to test this hypothesized effect. Following baseline observation, participants were instructed to refrain from exhibiting tics while watching videotapes, engaging in conversation, or while alone in a room with no activity. Observation continued following the suppression phase. Results of this experiment showed suppression of tics in almost one half of all sessions, with adults demonstrating suppression more frequently. Furthermore, results of this experiment failed to support a commonly held perception that following a period of voluntary suppression tics will rebound to a rate that will exceed the average rate of occurrence. PMID- 16046663 TI - Neuropsychological evaluation in the diagnosis and treatment of Tourette's syndrome. AB - The neurobiological basis of Tourette's syndrome is reviewed for the purpose of presenting a clinically relevant account of the neuropsychology of the disorder for the clinician who is behaviorally oriented. The neuropathology and neuropsychological deficits typically found in Tourette's are reviewed, and a neuropsychological test battery is described that can be used to help characterize the clinical presentation of the disorder. Although Tourette's syndrome is ultimately diagnosed by behavioral criteria, characterizing the cognitive deficits (or lack thereof) associated with the disorder is integral to fully appreciating the challenges posed by the disorder in any given case. The variety of cognitive deficits associated with Tourette's is reviewed to show the importance of the neuropsychological evaluation in differential diagnostic, therapeutic, and prognostic decisions. PMID- 16046664 TI - Tic or compulsion?: it's Tourettic OCD. AB - A subgroup of individuals suffering from obsessive-compulsive disorder (OCD) frequently present to treatment with an atypical yet distinguishable array of symptoms akin to both Tourette's disorder (TD) and OCD. These individuals often receive standard treatments for OCD (or less likely, TD) that fail to address the blended features of their presentation. It is argued that these individuals would be better served, both psychotherapeutically and pharmacologically, by the adoption of a Tourettic OCD (TOCD) conceptual framework. PMID- 16046665 TI - Adult bone marrow stem/progenitor cells (MSCs) are preconditioned by microenvironmental "niches" in culture: a two-stage hypothesis for regulation of MSC fate. AB - Mesenchymal stem cells (MSCs) are clonal, plastic adherent cells from bone marrow that can differentiate into various tissue lineages, including osteoblasts, adipocytes, chondrocytes, myoblasts, hepatocytes, and possibly even neural cells. Because MSCs are multipotent and their numbers are easily expanded in culture, there has been much interest in their clinical potential for tissue repair and gene therapy. Consequently, numerous studies have been carried out demonstrating the migration and multiorgan engraftment potential of MSCs in animal models and in human clinical trials. Understanding the mechanisms behind MSC cell fate determination is not easy, because the molecular processes that drive engraftment and differentiation are complex. Even in an in vitro system, the molecular cues necessary to induce differentiation are not easily identified or reproduced. In this Perspective, we emphasize the importance of microenvironmental factors in culture and suggest that MSC differentiation in vitro is regulated by a two-stage mechanism involving preconditioning by factors in the culture microenvironment followed by response to soluble differentiating factors. PMID- 16046666 TI - R7BP: a surprising new link between G proteins, RGS proteins, and nuclear signaling in the brain. AB - The regulators of G protein signaling (RGS proteins) bind directly to G protein alpha (Galpha) subunits in brain and other tissues to determine the strength, duration, and fidelity of neurotransmitter receptor signaling. A recent study shows, quite unexpectedly, that one class of RGS proteins [the R7 subfamily bound to Gbeta(5) (R7-Gbeta(5))] shuttles between the plasma membrane and the nucleus with assistance from a novel shuttle protein, R7BP. R7BP binds directly to R7 Gbeta(5) and the protein complex is tethered to the plasma membrane by addition of a lipid, palmitate, on R7BP. Removal of palmitate results in the translocation of the R7BP-R7-Gbeta(5) complex to the nucleus, presumably for nontraditional signaling functions. These findings suggest an entirely novel mechanism for regulating neurotransmitter signaling. That is, R7BP transduces signals directly from receptors and G proteins at the plasma membrane to the nucleus, and this plasma membrane-nuclear shuttling is controlled by reversible palmitoylation of R7BP. PMID- 16046667 TI - Teaching resources. Chromatin remodeling. AB - This Teaching Resource provides lecture notes and slides for a class covering chromatin remodeling mechanisms and is part of the course "Cell Signaling Systems: a Course for Graduate Students." The lecture begins with a discussion of chromatin organization and then proceeds to describe the process of chromatin remodeling through a review of chromatin remodeling complexes and methods used to study their function. PMID- 16046668 TI - Differential induction of connexins 26 and 30 in skin tumors and their adjacent epidermis. AB - Gap junctions (GJs) have been shown to play a role in tumor progression including a variety of keratinocyte-derived and non-keratinocyte-derived skin tumors. Here we show that the synthesis of the GJ proteins connexin 26 and connexin 30 (Cx26 and Cx30) is induced in keratinocyte-derived epithelial skin tumors whereas there is either no change or a downregulation of Cx43. Cx26, Cx30, and Cx43 are absent in non-epithelial skin tumors. Further, Cx26 and Cx30 are induced in the epidermis adjacent to malignant melanoma but absent in the epidermis adjacent to benign non-epithelial skin lesions (melanocytic nevi and angioma). The keratinocyte-derived skin tumors are very heterogeneous regarding the Cx26/Cx30 pattern in the epidermis at the periphery of the tumors. We did not observe any difference in the localization of the very similar proteins Cx26 and Cx30 but a variation in intensity of immunoreactivity. As the staining patterns of Cx26 and Cx30 antibodies are not identical to those of CK6, a marker for hyperproliferation, and CK17, a marker for trauma, we discuss that the induction of these gap junctional proteins exceeds a reflection of reactive hyperproliferative or traumatized epidermis. We further discuss the putative roles of these gap junctional proteins in tumor progression. PMID- 16046669 TI - Detection of myelination using a novel histological probe. AB - Current methods for myelin staining in tissue sections include both histological and immunohistochemical techniques. Fluorescence immunohistochemistry, which uses antibodies against myelin components such as myelin basic protein, is often used because of the convenience for multiple labeling. To facilitate studies on myelin, this paper describes a quick and easy method for direct myelin staining in rodent and human tissues using novel near-infrared myelin (NIM) dyes that are comparable to other well-characterized histochemical reagents. The near-infrared fluorescence spectra of these probes allow fluorescent staining of tissue sections in multiple channels using visible light fluorophores commonly used in immunocytochemistry. These dyes have been used successfully to detect normal myelin structure and myelin loss in a mouse model of demyelination disease. PMID- 16046670 TI - Improved localization of glucose-6-phosphate dehydrogenase activity in cells with 5-cyano-2,3-ditolyl-tetrazolium chloride as fluorescent redox dye reveals its cell cycle-dependent regulation. AB - Since the introduction of cyano-ditolyl-tetrazolium chloride (CTC), a tetrazolium salt that gives rise to a fluorescent formazan after reduction, it has been applied to quantify activity of dehydrogenases in individual cells using flow cytometry. Confocal laser scanning microscopy (CLSM) showed that the fluorescent formazan was exclusively localized at the surface of individual cells and not at intracellular sites of enzyme activity. In the present study, the technique has been optimized to localize activity of glucose-6-phosphate dehydrogenase (G6PD) intracellularly in individual cells. Activity was demonstrated in cultured fibrosarcoma cells in different stages of the cell cycle. Cells were incubated for the detection of G6PD activity using a medium containing 6% (w/v) polyvinyl alcohol, 5 mM CTC, magnesium chloride, sodium azide, the electron carrier methoxyphenazine methosulphate, NADP, and glucose-6-phosphate. Before incubation, cells were permeabilized with 0.025% glutaraldehyde. Fluorescent formazan was localized exclusively in the cytoplasm of fibrosarcoma cells. The amount of fluorescent formazan in cells increased linearly with incubation time when measured with flow cytometry and CLSM. When combining the Hoechst staining for DNA with the CTC method for the demonstration of G6PD activity, flow cytometry showed that G6PD activity of cells in S phase and G2/M phase is 27 +/- 4% and 43 +/- 4% higher, respectively, than that of cells in G1 phase. CLSM revealed that cells in all phases of mitosis as well as during apoptosis contained considerably lower G6PD activity than cells in interphase. It is concluded that posttranslational regulation of G6PD is responsible for this cell cycle-dependent activity. PMID- 16046671 TI - Genetic immunization: a new monoclonal antibody for the detection of BCL-6 protein in paraffin sections. AB - Genetic immunization can be combined with hybridoma technology to generate high affinity monoclonal antibodies (MAbs). A new anti-BCL-6 MAb (GI191E/A8) was produced by cloning full-length BCL-6 cDNA into a eukaryotic vector and delivering this into mouse epidermis using a helium gene gun. A comparative study was made of the specificity and the effects of formalin fixation on immunohistochemistry quality of GI191E/A8 and two other anti-BCL-6 MAbs. To evaluate its possible application to differential diagnosis of lymphomas, two tissue microarrays (89 diffuse large B-cell lymphomas and 24 B-cell chronic lymphocytic leukemia cases) were stained with GI191E/A8 and another anti-BCL-6 MAb produced by conventional means. Using GI191E/A8, the detection of BCL-6 protein was significantly increased, and its specificity was independent of formalin-fixation time. Using automatic quantified analysis, the correlation between the two anti-BCL-6 MAbs tested was identical in cases with overexpression or absence of BCL-6. In cases with intermediate BCL-6 protein expression, detection with GI191E/A8 was more sensitive. A significant association of higher BCL-6 expression and longer median overall survival times in diffuse large B-cell lymphomas was found. Using conventionally produced MAbs in the same patient group, the association was not significant. PMID- 16046672 TI - Application of heat-induced antigen retrieval to aldehyde-fixed fresh frozen sections. AB - We applied the heat-induced antigen retrieval (HIAR) to aldehyde-fixed fresh frozen sections based on a new approach (i.e., a rapid and complete immobilization of antigen followed by heating). Frozen sections were fixed with 10% formalin in 0.1 M cacodylate buffer (pH 7.4) containing 25 mM CaCl(2) for 30 min, or with 0.5% glutaraldehyde in 0.1 M phosphate buffer (pH 7.4) for 1 min at room temperature, and then autoclaved in 20 mM Tris-HCl buffer (pH 9.0) for 10 min at 120 C. Both fixatives yielded good tissue structure after autoclaving. In the sections fixed with formalin containing CaCl(2), 20 of 22 antigens located in the nucleus, cytoplasm, membranes, and extracellular matrix greatly recovered their antigenicity after autoclaving; only two antigens exhibited stronger immunoreaction in acetone-fixed fresh frozen sections than these sections. Heating also retrieved the immunoreactivity of at least 14 antigens in the sections fixed with glutaraldehyde. We used the similar procedures to localize ligand-free estrogen receptor alpha (ERalpha) and glucocorticoid receptors (GR). Mouse uterine cells exhibited almost the same nuclear ERalpha immunostaining regardless of the hormonal status in glutaraldehyde-fixed fresh frozen sections and unliganded GR was localized mainly in the nucleus of mouse hepatocytes in fresh frozen sections fixed with 20% formalin containing 50 or 75 mM CaCl(2) at 40 C, after autoclaving. These results demonstrate that HIAR is useful for the immunohistochemistry of many antigens in aldehyde-fixed fresh frozen sections. PMID- 16046673 TI - Research from tobacco industry affiliated authors: need for particular vigilance. PMID- 16046675 TI - Smoke-free laws do not harm profits: new global report. PMID- 16046676 TI - USA: PM's fouls come home to roost. PMID- 16046679 TI - Swedish Match: sucked into controversy, worldwide. PMID- 16046682 TI - Japanese spousal smoking study revisited: how a tobacco industry funded paper reached erroneous conclusions. AB - OBJECTIVES: To provide a participant's account of the development of a paper commissioned by the tobacco industry examining the reliability of self reported smoking status; to redress the distorted report of this Japanese spousal smoking study which evaluated the reliability and validity of self reported smoking status, and estimated confounding by diet and lifestyle factors. DESIGN: Repeated interviews on smoking status and its verification by environmental and biological markers for environmental tobacco smoke (ETS) exposure. SETTING: Urban wives in Osaka City and Sizuoka City, Japan PARTICIPANTS: Semi-random sampling of 200 wives in each city. From the Osaka subjects, 100 non-smoking wives were selected for the validity study. MAIN OUTCOME MEASURES: Kappa coefficient for reliability of self reported smoking status. Correlation coefficients between environmental nicotine concentration, cotinine in saliva and urine, and self reported smoking status. RESULTS: The kappa coefficient for the repeated interview was high suggesting sufficient reliability of the response. The proportion of self reported current smokers misclassified as non-smokers was equivalent to the misclassified self reported non-smokers. Ambient concentration of nicotine and personal exposure to nicotine correlated with each other and also with salivary cotinine and self reported ETS exposure but not with urinary cotinine/creatinine ratio (CCR). There was no major difference in diet and lifestyle related to husband's smoking status. CONCLUSION: Self reported smoking status by Japanese wives shows high reliability. It also shows high validity when verified by both nicotine exposure and salivary cotinine, but not by CCR. A previous report questioning the credibility of self reported smoking status, based on questionable CCR, could thus be of dubious validity. In addition, possible dietary and lifestyle confounding factors associated with smoking husbands were not demonstrable, a finding not reported previously. Using all the data from this project changes the conclusion of the previous published report. In addition to the distortion of scientific findings by a tobacco industry affiliated researcher, anti-smoking campaigners made attempts to intimidate and suppress scientific activities. These distortions of science should be counteracted. PMID- 16046685 TI - Changes in hospitality workers' exposure to secondhand smoke following the implementation of New York's smoke-free law. AB - OBJECTIVE: To assess the impact on hospitality workers' exposure to secondhand smoke of New York's smoke-free law that prohibits smoking in all places of employment, including restaurants, bars, and bowling facilities. DESIGN: Pre-post longitudinal follow up design. SETTINGS: Restaurants, bars, and bowling facilities in New York State. SUBJECTS: At baseline, 104 non-smoking workers in restaurants, bars, and bowling facilities were recruited with newspaper ads, flyers, and radio announcements. Of these, 68 completed a telephone survey and provided at least one saliva cotinine specimen at baseline. At three, six, and 12 month follow up studies, 47, 38, and 32 workers from the baseline sample of 68 completed a telephone survey and provided at least one saliva cotinine specimen. INTERVENTION: The smoke-free law went into effect 24 July 2003. MAIN OUTCOME MEASURES: Self reported sensory and respiratory symptoms and exposure to secondhand smoke; self administered saliva cotinine specimens. Analyses were limited to subjects in all four study periods who completed a telephone survey and provided at least one saliva cotinine specimen. RESULTS: All analyses were limited to participants who completed both an interview and a saliva specimen for all waves of data collection (n = 30) and who had cotinine concentrations < or = 15 ng/ml (n = 24). Hours of exposure to secondhand smoke in hospitality jobs decreased from 12.1 hours (95% confidence interval (CI) 8.0 to 16.3 hours) to 0.2 hours (95% CI -0.1 to 0.5 hours) (p < 0.01) and saliva cotinine concentration decreased from 3.6 ng/ml (95% CI 2.6 to 4.7 ng/ml) to 0.8 ng/ml (95% CI 0.4 to 1.2 ng/ml) (p < 0.01) from baseline to the 12 month follow up. The prevalence of workers reporting sensory symptoms declined from 88% (95% CI 66% to 96%) to 38% (95% CI 20% to 59%) (p < 0.01); there was no change in the overall prevalence of upper respiratory symptoms (p < 0.16). CONCLUSION: New York's smoke-free law had its intended effect of protecting hospitality workers from exposure to secondhand smoke within three months of implementation. One year after implementation, the results suggest continued compliance with the law. PMID- 16046686 TI - Smoking cessation treatment in primary care: prospective cohort study. AB - OBJECTIVE: To compare the characteristics of smokers who do and do not receive smoking cessation treatment in primary care. DESIGN: Prospective cohort study using practices registered with the pilot QRESEARCH database. SETTING: 156,550 patients aged 18 years and over from 39 general practices located within four strategic health authorities, representing the former Trent Region, UK. SUBJECTS: Patients registered with practices between 1 April 2001 and 31 March 2003 aged 18 years and over who were identified as smokers before the two year study period. OUTCOME: Prescription for smoking cessation treatment (nicotine replacement therapy (NRT) or bupropion) in the two year study period. VARIABLES: Age, sex, deprivation score, co-morbidity. RESULTS: Of the 29,492 patients recorded as current smokers at the start of the study period 1892 (6.4%) were given prescriptions for smoking cessation treatment during the subsequent two years. Of these, 1378 (72.8%) were given NRT alone, 406 (21.5%) bupropion alone, and 108 (5.7%) both treatments. Smokers were more likely to receive smoking cessation treatment if they lived in the most deprived areas (odds ratio (OR) for the most relative to the least deprived fifth, adjusted for sex, age, and co-morbidity, 1.50, 95% confidence interval (CI) 1.26 to 1.78), and if they were aged 25-74 years compared to 18-24 years or 75 and over. Smokers with co-morbidity were also more likely to receive smoking cessation treatment. Smokers were less likely to receive smoking cessation treatment if they were male (adjusted OR 0.68, 95% CI 0.62 to 0.75). CONCLUSION: The low proportion of smokers being prescribed these products strongly suggests that a major public health opportunity to prevent smoking related illness is being missed. PMID- 16046687 TI - Smoking, standard of living, and poverty in China. AB - OBJECTIVES: To analyse differences in smoking behaviour and smoking expenditures among low and high income households in China and the impact of smoking on standard of living of low income households in China. METHODS: About 3400 urban and rural households from 36 townships/districts in southwest China were interviewed in 2002. Cross tabulations and regression analysis were used to examine the differences in major household expenditures, including food, housing, clothing, and education between households with smokers and without smokers. RESULTS: Lower income households with smokers paid less per pack and smoked fewer cigarettes than higher income households with smokers. Poor urban households spent an average of 6.6% of their total expenditures on cigarettes; poor rural households spent 11.3% of their total expenditures on cigarettes. CONCLUSION: Reducing cigarette expenditures could release household resources to spend on food, housing, and other goods that improve living standards. PMID- 16046688 TI - Public health measures to reduce smoking prevalence in the UK: how many lives could be saved? AB - OBJECTIVE: To estimate the number of deaths that could be prevented in the UK by implementing population strategies to reduce smoking prevalence. DESIGN: A prospective analysis of future mortality using recent national smoking prevalence data and relative risks of mortality in current smokers, ex-smokers, and never smokers. POPULATION: Smokers in the UK. INTERVENTIONS: Population measures of proven effectiveness assumed to reduce smoking prevalence by 1 percentage point per year for 10 years, or alternatively by 13% over 19 years (1 percentage point per annum for seven years, 0.5 percentage point per annum for 12 years) as considered to be achievable in a recent report to the UK Chancellor of the Exchequer. MAIN OUTCOME MEASURE: Estimated deaths from smoking prevented in the 35-75 year age group. RESULTS: Reducing the prevalence of smoking by 1 percentage point each year for 10 years would prevent 69 049 deaths at ages between 35 and 74 years during that period. The model of reduction by 13% over 19 years would prevent 54 308 and 194 493 deaths in 10 and 19 years, respectively. Continued prevalence reductions at the current rate of 0.4 percentage points each year will prevent 23 192 deaths over 10 years. CONCLUSIONS: Full implementation of simple population measures to encourage smoking cessation could prevent substantial numbers of deaths in the UK. PMID- 16046689 TI - Do u smoke after txt? Results of a randomised trial of smoking cessation using mobile phone text messaging. AB - OBJECTIVES: To determine the effectiveness of a mobile phone text messaging smoking cessation programme. DESIGN: Randomised controlled trial SETTING: New Zealand PARTICIPANTS: 1705 smokers from throughout New Zealand who wanted to quit, were aged over 15 years, and owned a mobile phone were randomised to an intervention group that received regular, personalised text messages providing smoking cessation advice, support, and distraction, or to a control group. All participants received a free month of text messaging; starting for the intervention group on their quit day to assist with quitting, and starting for the control group at six months to encourage follow up. Follow up data were available for 1624 (95%) at six weeks and 1265 (74%) at six months. MAIN OUTCOME MEASURES: The main trial outcome was current non-smoking (that is, not smoking in the past week) six weeks after randomisation. Secondary outcomes included current non-smoking at 12 and 26 weeks. RESULTS: More participants had quit at six weeks in the intervention compared to the control group: 239 (28%) v 109 (13%), relative risk 2.20 (95% confidence interval 1.79 to 2.70), p < 0.0001. This treatment effect was consistent across subgroups defined by age, sex, income level, or geographic location (p homogeneity > 0.2). The relative risk estimates were similar in sensitivity analyses adjusting for missing data and salivary cotinine verification tests. Reported quit rates remained high at six months, but there was some uncertainty about between group differences because of incomplete follow up. CONCLUSIONS: This programme offers potential for a new way to help young smokers to quit, being affordable, personalised, age appropriate, and not location dependent. Future research should test these findings in different settings, and provide further assessment of long term quit rates. PMID- 16046690 TI - How Philip Morris built Marlboro into a global brand for young adults: implications for international tobacco control. AB - OBJECTIVE: To describe Philip Morris' global market research and international promotional strategies targeting young adults. METHODS: Analysis of previously secret tobacco industry documents. RESULTS: Philip Morris pursued standardised market research and strategic marketing plans in different regions throughout the world using research on young adults with three principle foci: lifestyle/psychographic research, brand studies, and advertising/communication effectiveness. Philip Morris identified core similarities in the lifestyles and needs of young consumers worldwide, such as independence, hedonism, freedom, and comfort. In the early 1990s Philip Morris adopted standardised global marketing efforts, creating a central advertising production bank and guidelines for brand images and promotions, but allowing regional managers to create regionally appropriate individual advertisements. CONCLUSIONS: Values and lifestyles play a central role in the global marketing of tobacco to young adults. Worldwide counter marketing initiatives, coupled with strong, coherent global marketing policies such as the Framework Convention on Tobacco Control, are needed to break associations between young adult values and tobacco brands. As globalisation promotes the homogenisation of values and lifestyles, tobacco control messages that resonate with young adults in one part of the world may appeal to young adults in other countries. Successful tobacco control messages that appeal to young people, such as industry denormalisation, may be expanded globally with appropriate tailoring to appeal to regional values. PMID- 16046691 TI - The perimetric boycott: a tool for tobacco control advocacy. AB - OBJECTIVES: To propose criteria to help advocates: (1) determine when tobacco related boycotts may be useful; (2) select appropriate targets; and (3) predict and measure boycott success. METHODS: Analysis of tobacco focused boycotts retrieved from internal tobacco industry documents websites and other scholarship on boycotts. RESULTS: Tobacco related boycotts may be characterised by boycott target and reason undertaken. Most boycotts targeted the industry itself and were called for political or economic reasons unrelated to tobacco disease, often resulting in settlements that gave the industry marketing and public relations advantages. Even a lengthy health focused boycott of tobacco industry food subsidiaries accomplished little, making demands the industry was unlikely to meet. In contrast, a perimetric boycott (targeting institutions at the perimeter of the core target) of an organisation that was taking tobacco money mobilised its constituency and convinced the organisation to end the practice. CONCLUSIONS: Direct boycotts of the industry have rarely advanced tobacco control. Perimetric boycotts of industry allies offer advocates a promising tool for further marginalising the industry. Successful boycotts include a focus on the public health consequences of tobacco use; an accessible point of pressure; a mutual interest between the target and the boycotters; realistic goals; and clear and measurable demands. PMID- 16046692 TI - Transforming the tobacco market: why the supply of cigarettes should be transferred from for-profit corporations to non-profit enterprises with a public health mandate. AB - Current tobacco control strategies seek primarily to decrease the demand for cigarettes through measures that encourage individuals to adopt healthier behaviours. These measures are impeded and undermined by tobacco corporations, whose profit drive compels them to seek to maintain and expand cigarette sales. Tobacco corporations seek to expand cigarette sales because they are for-profit business corporations and are obliged under law to maximise profits, even when this results in harm to others. It is not legally possible for a for-profit corporation to relinquish its responsibility to make profits or for it to temper this obligation with responsibilities to support health. Tobacco could be supplied through other non-profit enterprises. The elimination of profit driven behaviour from the supply of tobacco would enhance the ability of public health authorities to reduce tobacco use. Future tobacco control strategies can seek to transform the tobacco market from one occupied by for-profit corporations to one where tobacco is supplied by institutions that share a health mandate and will help to reduce smoking and smoking related disease and death. PMID- 16046693 TI - The effectiveness of television advertising campaigns on generating calls to a national Quitline by Maori. AB - OBJECTIVE: To examine the effectiveness of four mass media campaigns on calls to a national Quitline by Maori (the indigenous people of New Zealand). METHODS: Monthly Quitline call data and calls within one hour of a television commercial (TVC) being shown were analysed for the 2002-2003 period. Data on target audience rating points (TARPs) and expenditure on TVCs were also used (n = 2319 TVC placements). RESULTS: Maori were found to register with the Quitline at higher rates during the most intense six campaign months (15% more registrations compared to less intense months). The most effective campaign generated 115 calls per 100 TARPs by Maori callers within one hour of TVC airing (the "Every cigarette" campaign). A more Maori orientated campaign with both health and cultural themes generated 91 calls per 100 TARPs from Maori callers. For these two campaigns combined, the advertising cost per new registration with the Quitline by a Maori caller was NZD 30-48. Two second hand smoke campaigns that did not show the Quitline number were much less effective at 25 and 45 calls per 100 TARPs. CONCLUSIONS: These television advertising campaigns were effective and cost effective in generating calls to a national Quitline by Maori. Health authorities should continue to explore the use of both "threat appeal" style media campaigns and culturally appropriate campaigns to support Quitline use by indigenous peoples. PMID- 16046694 TI - New smoke-free environments legislation stimulates calls to a national Quitline. PMID- 16046696 TI - The human rights responsibilities of multinational tobacco companies. AB - This article explores various strategies which could be used to hold the tobacco industry accountable for human rights violations precipitated by its conduct. First, a brief overview of the international human rights regime and the tobacco related jurisprudence issued by human rights treaty bodies is provided. The article then explains how tobacco control advocates could promote more systematic consideration of governments' tobacco related human rights violations by reconceptualising the Framework Convention on Tobacco Control in the language of rights. The feasibility of using the existing human rights framework to target the tobacco industry directly is analysed with the conclusion that this approach has serious limitations. Emerging human rights norms, which have greater potential to affect the industry's conduct, are presented. Finally, given the questionable authoritativeness of these norms, alternative ways that they could be employed to hold tobacco companies accountable for the rights related consequences of their activities are proposed. PMID- 16046697 TI - International trade agreements: a threat to tobacco control policy. AB - International covenants establish a role for governments in ensuring the conditions for human health and wellbeing, which has been recognised as a central human right. International trade agreements, conversely, prioritize the rights of corporations over health and human rights. International trade agreements are threatening existing tobacco control policies and restrict the possibility of implementing new controls. This situation is unrecognised by many tobacco control advocates in signatory nations, especially those in developing countries. Recent agreements on eliminating various trade restrictions, including those on tobacco, have expanded far beyond simply international movement of goods to include internal tobacco distribution regulations and intellectual property rules regulating advertising and labelling. Our analysis shows that to the extent trade agreements protect the tobacco industry, in itself a deadly enterprise, they erode human rights principles and contribute to ill health. The tobacco industry has used trade policy to undermine effective barriers to tobacco importation. Trade negotiations provide an unwarranted opportunity for the tobacco industry to assert its interests without public scrutiny. Trade agreements provide the industry with additional tools to obstruct control policies in both developed and developing countries and at every level. The health community should become involved in reversing these trends, and help promote additional measures to protect public health. PMID- 16046698 TI - Ethical and legal aspects of global tobacco control. AB - On 28 February 2005, the Framework Convention on Tobacco Control came into force as a result of at least 40 countries becoming State Parties through ratification of this first ever health treaty sponsored by the World Health Organization. This article discusses the bioethical, trade, and legal aspects of global tobacco control. Special emphasis is given to globalisation of tobacco use and the challenges it poses to sovereign nations. It also advocates a bioethical basis in the pursuit of global solutions to expanding tobacco use. PMID- 16046699 TI - "Not safe" is not enough: smokers have a right to know more than there is no safe tobacco product. AB - The right to health relevant information derives from the principles of autonomy and self direction and has been recognised in international declarations. Providing accurate health information is part of the basis for obtaining "informed consent" and is a recognised component of business ethics, safety communications, and case and product liability law. Remarkably, anti-tobacco and pro-tobacco sources alike have come to emphasise the message that there is "no safe cigarette" or "no safe tobacco product". We propose that the "no safe" message is so limited in its value that it represents a violation of the right to health relevant information. There is a need to go beyond saying, "there is no safe tobacco product" to indicate information on degree of risks. The "no safe tobacco" message does not contradict, for example, the mistaken belief that so called light or low tar cigarettes are safer choices than higher tar cigarettes. We encourage a kind of "rule utilitarian" ethical position in which the principle of truth telling is observed while trying to produce the greatest good for the greatest number of people. Although harm reduction approaches to easing the burden of tobacco related diseases are founded on science based comparative risk information, the right to health information is independently related to the need to promote health literacy. This right should be respected whether or not harm reduction policies are judged advisable. PMID- 16046700 TI - Individual rights advocacy in tobacco control policies: an assessment and recommendation. AB - Efforts to control environmental tobacco smoke (ETS) could be assisted if the tobacco control movement gave greater emphasis to the issue of individual rights. Benefits that may accrue from the promotion of a clear individual rights perspective in tobacco control include adding coherence to the tobacco control advocacy agenda and winning support from those who may have been concerned about loss of personal freedom, excessive governmental power, use of social coercion, or the rights of smokers. Risks also attend to such a policy. It might inadvertently assist the tobacco industry, stir resistance to ETS limitation efforts, or confuse tobacco control supporters. On balance, though, liabilities are outweighed by the ethical and operational merits in tobacco control of a heightened pro-individual rights stance. PMID- 16046701 TI - Framing tobacco control efforts within an ethical context. AB - Public health efforts to promote tobacco control are not performed within a vacuum. They are subject to interpretation and misinterpretation by consumers and policymakers based largely upon the initial framing of the issues. This paper notes how the tobacco industry has established a particular frame that it is the protector of individual rights and that the public health community is trying to eliminate those rights. This paper then shows how the public health community uses metaphors that may unintentionally support this framing and suggests that by reframing public health efforts in accordance with core ethical principles, the public health community can create more positive messages. A public health ethical framework is proposed to examine how the application of the principles can influence the tobacco control movement. Through the increased use of ethics in tobacco control, the public health community may be better positioned to claim the high road as the protector of the public's interests. PMID- 16046702 TI - Social movements and human rights rhetoric in tobacco control. AB - After achieving breathtaking successes in securing state and local restrictions on smoking in public places and restricting youth access to tobacco products, the tobacco movement faces difficult decisions on its future strategic directions. The thesis of this article is that the tobacco control movement is at a point of needing to secure its recent successes and avoiding any public retrenchment. To do so requires rethinking the movement's strategic direction. We use the familiar trans-theoretical model of change to describe where the movement is currently and the threats it faces. The new tobacco control strategy should encompass a focus on voluntary non-smoking strategies, use human rights rhetoric to its advantage, and strengthen the public health voice to be more effective in political battles. In developing a new strategy, tobacco control advocates need to build a social movement based on a more forceful public health voice, along with the strategic use of human rights rhetoric, to focus on the power of voluntary non-smoking efforts. Using human rights rhetoric can help frame the movement in ways that have traditionally appealed to the American public. Perhaps more importantly, doing so can help infuse the tobacco control movement with a broader sense of purpose and mission. PMID- 16046703 TI - Ensuring smokers are adequately informed: reflections on consumer rights, manufacturer responsibilities, and policy implications. AB - The right to information is a fundamental consumer value. Following the advent of health warnings, the tobacco industry has repeatedly asserted that smokers are fully informed of the risks they take, while evidence demonstrates widespread superficial levels of awareness and understanding. There remains much that tobacco companies could do to fulfil their responsibilities to inform smokers. We explore issues involved in the meaning of "adequately informed" smoking and discuss some of the key policy and regulatory implications. We use the idea of a smoker licensing scheme-under which it would be illegal to sell to smokers who had not demonstrated an adequate level of awareness-as a device to explore some of these issues. We also explore some of the difficulties that addiction poses for the notion that smokers might ever voluntarily assume the risks of smoking. PMID- 16046704 TI - Combination diet and exercise interventions for the treatment of dyslipidemia: an effective preliminary strategy to lower cholesterol levels? AB - At present, dyslipidemia is most commonly treated with drug therapy. However, because safety concerns regarding the use of pharmaceutical agents have arisen, a need for alternative nonpharmacological therapies has become increasingly apparent. The National Cholesterol Education Program (NCEP) Adult Treatment Panel III (ATP III) recommends lifestyle therapies, which include a combination of diet and exercise modifications, in place of drug treatment for patients who fall into an intermediate range of coronary heart disease (CHD) risk. This review examined the cholesterol lowering efficacy of the following 2 NCEP-recommended combination therapies: 1) low saturated fat diets combined with exercise, and 2) nutritional supplementation, i.e., fish oil, oat bran, or plant sterol supplementation, combined with exercise, in the treatment of dyslipidemia. Combination therapies are particularly advantageous because diet and exercise elicit complementary effects on lipid profiles. More specifically, diet therapies, with some exceptions, lower total (TC) and LDL cholesterol (LDL-C) concentrations, whereas exercise interventions increase HDL cholesterol (HDL-C) while decreasing triglyceride (TG) levels. With respect to specific interventions, low saturated fat diets combined with exercise lowered TC, LDL-C, and TG concentrations by 7 18, 7-15, and 4-18%, respectively, while increasing HDL-C levels by 5-14%. Alternatively, nutritional supplements combined with exercise, decreased TC, LDL C, and TG concentrations by 8-26, 8-30, and 12-39%, respectively, while increasing HDL-C levels by 2-8%. These findings suggest that combination lifestyle therapies are an efficacious, preliminary means of improving cholesterol levels in those diagnosed with dyslipidemia, and should be implemented in place of drug therapy when cholesterol levels fall just above the normal range. PMID- 16046705 TI - Folate deficiency-induced hyperhomocysteinemia attenuates, and folic acid supplementation restores, the functional activities of rat coagulation factors XII, X, and II. AB - Hyperhomocysteinemia (HH) constitutes a risk marker for thrombosis, but the pathophysiological mechanisms in thrombus formation are still unresolved. We investigated the influence of HH on single coagulation factor functions and evaluated the platelet GpIIb/IIIa receptor function in HH-induced changes in whole-blood coagulation profiles (WBCP). Three groups of 12 rats were investigated: control rats, folate deficient-HH (FD-HH) rats, and treated rats. Plasma total homocysteine was 7.1 micromol/L in controls, 31.3 micromol/L in FD HH rats, and 7.6 micromol/L in treated rats. Factor (F) II:C, FX:C, and FXII:C were reduced in FD-HH rats compared with controls and normalized in treated rats (P < 0.05). FVII:C activity did not differ among the groups. Factor VIII:C activity was greater in FD-HH rats than in controls (P < 0.05). Blockage of the platelet GpIIb/IIIa receptor by Integrilin (Schering-Plough A/S) did not abolish the FD-HH-induced increase in whole-blood coagulation velocity, irrespective of the dosage of Integrilin. In conclusion, FD-HH reduced the functional activities of FXII:C, FX:C and FII:C, whereas FVII:C was unchanged and FVIII:C increased. These findings may partially explain the prolonged initiation phase of WBCP in FD HH rats. The changes in single coagulation factor functions and WBCPs in FD-HH rats were reversed by treatment with folic acid. PMID- 16046706 TI - Palmitate activates the NF-kappaB transcription factor and induces IL-6 and TNFalpha expression in 3T3-L1 adipocytes. AB - Fatty acids and their metabolites regulate gene expression and immunological pathways. Furthermore, obese individuals frequently have increased circulating fatty acid concentrations, and localized inflammation in adipose tissue may facilitate the systemic inflammation associated with the insulin resistance of obesity. Although palmitate induces inflammation (i.e., activates proinflammatory pathways) in myotubes, the effects of fatty acids on inflammatory processes in adipocytes have not been established. Therefore, we examined the potential for palmitate, laurate, and docosahexaenoic acid (DHA) to modulate inflammation in 3T3-L1 adipocytes. Palmitate, but not DHA or laurate, induced nuclear factor kappaB (NF-kappaB)-driven luciferase activity and interleukin-6 (IL-6) expression (P < 0.05). Inhibition of fatty acyl Co-A synthase (FACS) with triacsin C suppressed palmitate-induced NF-kappaB activation (P < 0.05), but caused an additive increase in palmitate-induced IL-6 expression (P < 0.05). Disrupting mitogen-activated protein kinase/Erk kinase (MEK) and protein kinase C (PKC) activity with U0126 and Bisindolylmaleimide (Bis), respectively, suppressed palmitate-induced IL-6 expression (P < 0.05), but had no effect on NF-kappaB reporter gene activity (P > 0.05). However, the phosphoinositide-3 kinase (PI3K) inhibitor, wortmannin, alone and additively with palmitate, activated the NF kappaB reporter gene and induced IL-6 expression (P < 0.05). Palmitate also induced the mRNA expression of tumor necrosis factor alpha (TNFalpha) (P < 0.05), but the increase in mRNA abundance was not reflected in a greater protein concentration in the media (P > 0.05). These data indicate that palmitate induces inflammation in adipocytes, and that this is not a generalized effect of all SFA. Furthermore, PI3K may act constitutively to suppress inflammation. Consequently, inhibition of this enzyme may promote and exacerbate the inflammation in adipose tissue that is associated with obesity and insulin resistance. PMID- 16046707 TI - Vitamin E increases production of vasodilator prostanoids in human aortic endothelial cells through opposing effects on cyclooxygenase-2 and phospholipase A2. AB - Impairment of endothelium-dependent vasodilation is associated with the initiation and development of atherosclerosis. Vasodilator prostanoids constitute a protective mechanism in maintaining normal vasomotor function. In the current study, we determined the effect of in vitro vitamin E supplementation at physiologically relevant concentrations (10-60 micromol/L) on the production of the vasodilator prostanoids prostaglandin I(2) (PGI(2); prostacyclin) and prostaglandin E(2)(PGE(2)) by human aortic endothelial cells (HAECs) as well as its underlying mechanism. Results showed that vitamin E dose dependently (10-40 micromol/L) increased the production of both prostanoids by HAECs. This was associated with a dose-dependent (10-40 micromol/L) upregulation of cytosolic phospholipase A(2) (cPLA(2)) expression and arachidonic acid release. In contrast, vitamin E dose dependently (10-60 micromol/L) inhibited cyclooxygenase (COX) activity but did not affect the expression of either COX-1 or COX-2, indicating that the effect of vitamin E on COX activity was post-translational. Thus, vitamin E had opposing effects on the 2 key enzymes in prostanoid biosynthesis; at the concentrations used in this study, this resulted in a net increase in the production of vasodilator prostanoids. The vitamin E-induced increase in PGI(2) and PGE(2) production may contribute to its suggested beneficial effect in preserving endothelial function. PMID- 16046708 TI - Increasing the protein:carbohydrate ratio in a high-fat diet delays the development of adiposity and improves glucose homeostasis in mice. AB - Dietary fat is considered an important contributing factor in the obesity epidemic, and high-fat diets are used widely to induce obesity and diabetes related traits in susceptible rodent strains. Little attention, however, is usually paid to the interaction of fat with the other macronutrients. The aim of this study, therefore, was to investigate the effects of high-fat, isoenergetic diets with different protein:carbohydrate (CHO) ratios on obesity, energy metabolism, and glucose homeostasis in mice. Male adult C57BL/6J mice consumed ad libitum for 10 wk a control diet (41:42:17 ratio of CHO:protein:fat, 15.5 kJ/g) or 2 different high-fat diets: high carbohydrate (HC; 41:16:43, 17.7 kJ/g) or low carbohydrate (LC; 11:45:44, 17.5 kJ/g). Body weight and fat gains were rapid and were greater in HC mice than in other groups due to an initial pronounced hyperphagia and subsequent passive overconsumption. Weight and fat gains were less in LC mice but still greater than in controls. Energy expenditure was not affected by the diets, and total energy intake explained 84% of the variation in final body weight. The respiratory quotient was lower in LC mice than in other groups, indicating high fat oxidation rates due to the LC diet. Blood glucose was lower and insulin sensitivity greater in LC mice than in HC mice. We conclude that increasing the protein:CHO ratio in a high-fat diet delays but does not prevent the development of adiposity. However, glucose homeostasis was improved in LC mice, indicating that a combination of high fat and high CHO is responsible for the development of metabolic syndrome-related traits in mice. PMID- 16046709 TI - Curcumin blocks interleukin-1 (IL-1) signaling by inhibiting the recruitment of the IL-1 receptor-associated kinase IRAK in murine thymoma EL-4 cells. AB - Curcumin is a dietary compound with diverse anti-inflammatory and anticarcinogenic effects in several experimental models. A mechanism by which curcumin exerts these actions might be the direct modification of protein thiols, thereby altering the activity of the affected proteins. An early event in inflammatory signaling cascades is the recruitment of the interleukin-1 (IL-1) receptor-associated kinase (IRAK) to the IL-1 receptor (IL-1RI) upon stimulation with IL-1. IRAK recruitment was shown recently to be inhibited by agents that modify thiols of IRAK. We asked, therefore, whether IRAK is also a target for curcumin. Curcumin indeed blocked IRAK thiols in a murine T-cell line stably overexpressing IRAK (EL-4(IRAK)), which resulted in the inhibition of IRAK recruitment to the IL-1RI and phosphorylation of IRAK and IL-1RI-associated proteins. Inhibitory effects were not reversible by thiol-reducing agents. Thus, modification by curcumin did not occur by oxidation but rather by alkylation, as is typical for electrophilic compounds reacting as Michael addition acceptors. The block in one of the earliest events in the IL-1 signaling cascade can explain the often observed inhibition of IL-1-mediated signaling steps by curcumin further downstream. Hence, thiol modification might be a crucial step in the anti inflammatory functions of curcumin. PMID- 16046710 TI - Transcriptome analysis of human colon Caco-2 cells exposed to sulforaphane. AB - Sulforaphane (SF), a dietary phytochemical obtained from broccoli, has been implicated in several physiological processes consistent with anticarcinogenic activity, including enhanced xenobiotic metabolism, cell cycle arrest, and apoptosis. In this study, we report changes in global gene expression in Caco-2 cells exposed to physiologically appropriate concentrations of SF, through the use of replicated Affymetrix array and RT-PCR experiments. After exposure to 50 micromol/L SF, 106 genes exhibited a >2-fold increase in expression and 63 genes exhibited a >2-fold decrease in expression. There were fewer changes in gene expression at lower SF concentrations. The majority of these genes had not previously been shown to be modulated by SF, suggesting novel mechanisms of possible anticarcinogenic activity, including induction of differentiation and modulation of fatty acid metabolism. The changes in the expression of 10 of these genes, together with 4 additional genes of biological interest, were further quantified in independent studies with RT-PCR. These genes include several that have recently become associated with carcinogenesis, such as Kruppel-like factor (KLF)4, a gut-enriched transcription factor associated with induction of differentiation and reduction in cellular proliferation; DNA (cytosine-5-) methyltransferase 1, associated with methylation; and alpha-methylacyl-CoA racemase (AMACR), a marker associated with the development of colon and prostate cancer. The expression of 5 of these genes [caudal type homeo box transcription factor 2 (CDX-2), KLF4, KLF5, cyclin-dependent kinase inhibitor 1A (p21), and AMACR] was additionally studied after in vitro exposure to SF of surgically resected healthy and cancerous colon tissue from each of 3 patients. The study suggests the complex effects that SF has on gene expression and highlights several potential mechanisms by which the consumption of broccoli may reduce the risk of carcinogenesis. PMID- 16046711 TI - Low-protein diets reduce PKAalpha expression in islets from pregnant rats. AB - We investigated the effect of protein restriction on insulin secretion and the expression of protein kinase (PK)Aalpha and PKCalpha in islets from control and pregnant rats. Adult control nonpregnant (CN) and control pregnant (CP) rats were fed a normal-protein diet (17%), whereas low-protein nonpregnant (LPN) and low protein pregnant (LPP) rats were fed a low-protein diet (6%) for 15 d. In the presence of 2.8 and 8.3 mmol glucose/L, insulin secretion by islets of CP rats was higher than that by islets of CN rats. Compared with the CN groups, insulin secretion by islets of LPN rats was lower with 8.3 but not with 2.8 mmol glucose/L. The insulin secretion by islets of LPP rats was higher than by LPN rats at both glucose concentrations. IBMX (1 mmol/L), a phosphodiesterase inhibitor, increased insulin secretion by islets from pregnant rats, and this effect was greater in islets of CP rats than in LPP rats. Forskolin (0.01-100 micromol/L), a stimulator of adenylyl cyclase, increased insulin secretion only in islets of CN and CP rats, with a higher 50% effective concentration in islets of CP rats compared with CN rats. The insulin secretion induced by phorbol 12 myristate 13-acetate (a stimulator of PKC) was higher in islets of LPN and LPP rats than in the respective controls, especially at 8.3 mmol glucose/L. PKAalpha, but not PKCalpha, expression was lower in islets of rats fed low protein than in the controls, regardless of the physiological status of the rats. All endocrine cells of the islets, including beta-cells, expressed the PKAalpha isoform. The cytoplasmic distribution of this enzyme in beta-cells was not modified by pregnancy and/or protein restriction. In conclusion, our results indicate that the response of islets from rats fed low protein during pregnancy is similar to that of control rats, at least for physiologic glucose concentration. However, the decreased response to IBMX and forskolin indicates decreased production and/or sensitivity to cAMP; this was associated with a decrease in PKA expression, which may result in lower PKA activity. PMID- 16046712 TI - Vegetables affect the expression of genes involved in anticarcinogenic processes in the colonic mucosa of C57BL/6 female mice. AB - There is abundant epidemiological evidence that vegetable consumption decreases colorectal cancer (CRC) risk. However, the molecular targets in the genome are mostly unknown. The present study investigated the effects of vegetable consumption on gene expression in the colon mucosa of female C57Bl/6 mice using cDNA microarray technology. Mice were fed one of 8 diets: a control diet containing no vegetables (diet 1); a diet containing 100 g/kg (diet 2, 10% dose), 200 g/kg (diet 3, 20% dose), or 400 g/kg (diet 4, 40% dose) of a vegetable mixture; or a diet containing 70 g/kg of cauliflower (diet 5, 7% dose), 73 g/kg of carrots (diet 6, 7.3% dose), 226 g/kg of peas (diet 7, 22.6% dose); or 31 g/kg of onions (diet 8, 3.1% dose). The vegetable mixture used in diets 2 to 4 consisted of the 4 individual vegetables used in diets 5 to 8: cauliflower (30% wet wt), carrots (30% wet wt), peas (30% wet wt), and onions (10% wet wt). To assess gene expression changes, colonic mucosal cells were collected after the mice were killed. Total RNA was isolated and microarray technology was used to measure the expression levels of 602 genes simultaneously. For 39 genes, significant dose-dependent effects were found, although in general the relations were not linear. For 15 genes, the altered expression could indeed explain reduced cancer risk at various stages of CRC development. Eleven genes were modulated by the vegetable mixture as well as by one or more of the individual vegetables. For 7 of the genes, the modulation by the mixture was due to the effect of a particular vegetable. These genes are of particular interest because they were consistently affected and could be involved in the prevention of CRC by vegetable consumption. PMID- 16046713 TI - Complementary feeding with cow's milk alters sleeping metabolic rate in breast fed infants. AB - Although it is widely accepted that energy expenditure in infants is a function of feeding pattern, the mechanism behind this is not well understood. The objectives of this observational study were as follows: 1) to compare minimal observable energy expenditure (MOEE) between 2 subgroups of breast-fed infants, a BM group in which breast milk was the only source of milk and a BCM group given cow's milk in addition to breast milk; and 2) to identify potential mediators of a feeding pattern effect. For this purpose, infants were classified by feeding group on the basis of a mother's recall. Respiration calorimetry was used to measure MOEE in 62 infants (n = 35 BM, n = 27 BCM) aged 8.7 mo in Pelotas, southern Brazil. Breast-milk intake was measured using deuterium oxide, complementary food intake by 1-d food weighing, total energy expenditure and total body water using doubly labeled water; anthropometric indices were calculated. MOEE was 1672 +/- 175 kJ/d in BM compared with 1858 +/- 210 kJ/d in BCM infants (P < 0.001). Mass-specific MOEE was 201 +/- 24.6 and 216 +/- 31.9 kJ/(kg . d) in BM and BCM infants, respectively (P = 0.041). MOEE (kJ/d) was mediated by protein intake and fat-free mass (R(2) = 41.4%). We conclude that complementary feeding with cow's milk alters the sleeping metabolic rate in breast-fed infants. These findings deserve attention in relation to "metabolic programming" and the development of obesity later in life. PMID- 16046714 TI - Fructooligosaccharides and fiber partially prevent the alterations in fecal microbiota and short-chain fatty acid concentrations caused by standard enteral formula in healthy humans. AB - The intestinal microbiota are important during enteral tube feeding because they exert colonization resistance and produce SCFAs. However, the effect of the enteral formula composition on major bacterial groups of the microbiota has not been clearly defined. The aim of this study was to investigate the effect of enteral formulas with and without prebiotic fructooligosaccharides (FOS) and fiber on the fecal microbiota and SCFAs. Healthy subjects (n = 10; 4 men, 6 women) consumed both a standard enteral formula and one containing FOS (5.1 g/L) and fiber (8.9 g/L) as a sole source of nutrition for 14 d in a randomized, double-blind, crossover trial with a 6-wk washout phase. Fecal samples were collected at the start and end of each formula phase, and were analyzed for major bacterial groups and SCFA concentrations using fluorescent in situ hybridization and GLC, respectively. Although there were reductions in total fecal bacteria due to both formula treatments, concentrations were higher after the FOS/fiber formula period compared with the standard formula period (11.2 +/- 0.2 vs. 11.0 +/- 0.2 log(10) cells/g, P = 0.005). The FOS/fiber formula increased bifidobacteria (P = 0.004) and reduced clostridia (P = 0.006). Compared with the standard formula, the FOS/fiber formula resulted in higher concentrations of total SCFA (332.4 +/- 133.8 vs. 220.1 +/- 124.5 micromol/g, P = 0.022), acetate (219.6 +/- 96.3 vs. 136.8 +/- 74.5 micromol/g, P = 0.034) and propionate (58.4 +/ 37.4 vs. 35.6 +/- 25.5 micromol/g, P = 0.02). This study demonstrates that standard enteral formula leads to adverse alterations to the fecal microbiota and SCFA concentrations in healthy subjects, and these alterations are partially prevented by fortification of the formula with FOS and fiber. PMID- 16046715 TI - Dietary protein and exercise have additive effects on body composition during weight loss in adult women. AB - This study examined the interaction of 2 diets (high protein, reduced carbohydrates vs. low protein, high carbohydrates) with exercise on body composition and blood lipids in women (n = 48, approximately 46 y old, BMI = 33 kg/m(2)) during weight loss. The study was a 4-mo weight loss trial using a 2 x 2 block design (Diet x Exercise). Diets were equal in total energy (7.1 MJ/d) and lipids ( approximately 30% energy intake) but differed in protein content and the ratio of carbohydrate:protein at 1.6 g/(kg . d) and <1.5 (PRO group) vs. 0.8 g/(kg . d) and >3.5 (CHO group), respectively. Exercise comparisons were lifestyle activity (control) vs. a supervised exercise program (EX: 5 d/wk walking and 2 d/wk resistance training). Subjects in the PRO and PRO + EX groups lost more total weight and fat mass and tended to lose less lean mass (P = 0.10) than the CHO and CHO + EX groups. Exercise increased loss of body fat and preserved lean mass. The combined effects of diet and exercise were additive for improving body composition. Serum lipid profiles improved in all groups, but changes varied among diet treatments. Subjects in the CHO groups had larger reductions in total cholesterol and LDL cholesterol, whereas subjects in the PRO groups had greater reductions in triacylglycerol and maintained higher concentrations of HDL cholesterol. This study demonstrated that a diet with higher protein and reduced carbohydrates combined with exercise additively improved body composition during weight loss, whereas the effects on blood lipids differed between diet treatments. PMID- 16046716 TI - Grape polyphenols exert a cardioprotective effect in pre- and postmenopausal women by lowering plasma lipids and reducing oxidative stress. AB - To evaluate the effects of grape polyphenols on plasma lipids, inflammatory cytokines, and oxidative stress, 24 pre- and 20 postmenopausal women were randomly assigned to consume 36 g of a lyophilized grape powder (LGP) or a placebo for 4 wk. The LGP consisted of 92% carbohydrate and was rich in flavans, anthocyanins, quercetin, myricetin, kaempferol, and resveratrol. After a 3-wk washout period, subjects were assigned to the alternate treatment for an additional 4 wk. The placebo consisted of an equal ratio of fructose and dextrose and was similar in appearance and energy content (554 kJ) to LGP. Plasma triglyceride concentrations were reduced by 15 and 6% in pre- and postmenopausal women, respectively (P < 0.01) after LGP supplementation. In addition, plasma LDL cholesterol and apolipoproteins B and E were lower due to LGP treatment (P < 0.05). Further, cholesterol ester transfer protein activity was decreased by approximately 15% with intake of LGP (P < 0.05). In contrast to these beneficial effects on plasma lipids, LDL oxidation was not modified by LGP treatment. However, whole-body oxidative stress as measured by urinary F(2)-isoprostanes was significantly reduced after LGP supplementation. LGP also decreased the levels of plasma tumor necrosis factor-alpha, which plays a major role in the inflammation process. Through alterations in lipoprotein metabolism, oxidative stress, and inflammatory markers, LGP intake beneficially affected key risk factors for coronary heart disease in both pre- and postmenopausal women. PMID- 16046717 TI - Early childhood development interventions and cognitive development of young children in rural Vietnam. AB - Little is known about the long-term benefits of interventions that aim to promote early childhood development programs. The goal of this research was to determine whether an early childhood development intervention added to a nutrition intervention during preschool ages had lasting effects on the cognitive development of school-age children in communes of Thanh Hoa province in rural Vietnam. The study focused on a total of 313 children aged 6.5-8.5 y (grades 1 and 2 in primary school) in 2 communes that were exposed to nutrition intervention or nutrition and early childhood development (ECD) intervention from 1999 to 2003. Measurements of height and cognitive test scores (Raven's Progressive Matrices Test) were collected from the children; household characteristics were determined by interviews with mothers. Longitudinal analysis was performed by integrating the data with that collected from the same children in past surveys. Significant effects of the ECD intervention compared with the nutrition intervention were detected. The beneficial effect of ECD intervention on the cognitive test scores was large for the most nutritionally challenged children whose height-for-age Z-scores declined or remained in the stunted range. The findings help provide useful insights into the development of an effective integrated model of ECD and nutrition intervention for children in rural Vietnam. PMID- 16046718 TI - Homeless youth in Toronto are nutritionally vulnerable. AB - This study was undertaken to characterize nutritional vulnerability among a sample of homeless youth in downtown Toronto. Interviews were conducted with 261 homeless youth (149 male, 112 female), recruited from drop-in centers and outdoor locations. Information about current living circumstances, nutrition and health related behaviors, and 24-h dietary intake recalls were collected, and height, weight, triceps skinfold thickness, and mid-upper arm circumference were measured. A second 24-h dietary intake recall was conducted with 195 youth. Youth's energy intakes approximated the requirements for a very sedentary lifestyle; 7% were underweight and 22% were overweight or obese. Over half of the youth had inadequate intakes of folate, vitamin A, vitamin C, magnesium, and zinc; in addition, more than half of females had inadequate vitamin B-12 and iron intakes. Most youth got food from more than one source in the course of a day: 74% of males and 75% of females purchased food; 48% of males and 51% of females obtained food from charitable meal programs; 47% of males and 75% of females received food from strangers or acquaintances; and 10% of males and 6% of females stole food or took it from the garbage. Compared to a sample of 114 domiciled youth from the 1997-1998 Ontario Food Survey, males had lower energy and nutrient intakes and females had lower intakes of most nutrients. PMID- 16046719 TI - Butter naturally enriched in conjugated linoleic acid and vaccenic acid alters tissue fatty acids and improves the plasma lipoprotein profile in cholesterol-fed hamsters. AB - Butter, which is naturally enriched in cis-9, trans-11 conjugated linoleic acid (rumenic acid; RA) and vaccenic acid (VA), has been shown to be an effective anticarcinogen in studies with animal models; however, there has been no examination of the effects of a naturally derived source of VA and RA on atherosclerosis-related biomarkers. The current study was designed to determine the effect of a diet containing VA/RA-enriched butter on plasma lipoproteins and tissue fatty acid profiles in cholesterol-fed hamsters. Male Golden Syrian hamsters were fed diets containing 0.2% cholesterol and 20% added fat as: 1) Control, 20% standard butter (CT); 2) 5% standard butter + 15% VA/RA-enriched butter (EB); 3) 15% standard butter + 5% partially-hydrogenated vegetable oil (VO). After 4 wk, plasma lipoproteins were isolated, cholesterol quantified, and tissue fatty acid profiles determined. Tissue concentrations of VA and RA were increased by consumption of the EB diet compared with both the CT and VO diets, whereas the VO diet increased their concentration compared with the CT diet only. Total and LDL cholesterol concentrations were significantly reduced in hamsters fed EB and VO compared with CT, whereas VLDL cholesterol concentrations were reduced in hamsters fed EB compared with those fed CT and VO. HDL cholesterol concentrations did not differ among treatments. The ratio of potentially atherogenic lipoproteins [VLDL + intermediate density lipoproteins (IDL) + LDL] to antiatherogenic HDL was significantly lower in hamsters fed VA/RA-enriched butter (0.60) than in those fed either control diet (1.70) or the diet containing partially hydrogenated vegetable oil (1.04). Thus, increasing the VA/RA concentration of butter results in a plasma lipoprotein cholesterol profile that is associated with a reduced risk of atherosclerosis. PMID- 16046720 TI - Diet and age affect intestinal morphology and large bowel fermentative end product concentrations in senior and young adult dogs. AB - The objective of this study was to determine the effects of age and diet on intestinal morphology and large bowel fermentative end-product concentrations in healthy dogs. Small intestinal villus width, height, and area, and small intestinal and colonic crypt depth were measured. Large bowel digesta samples were analyzed for ammonia, SCFAs, and branched-chain fatty acids (BCFAs). SCFAs are considered to be beneficial fermentative end-products in the intestine because they exert trophic effects on intestinal cells. Twelve senior (age = 11.1 y +/- 0.6 at baseline; 6 male, 6 female) and 12 young adult (age = 8 wk old at baseline; 6 male, 6 female) beagles were randomly assigned to 1 of 2 dietary treatments, an animal product-based diet (APB) and a plant product-based diet (PPB). Diets were fed for 12 mo. Jejunal (P = 0.03) and ileal (P = 0.02) villus height, and duodenal (P = 0.04) villus width were greater for dogs consuming the PPB diet. Young dogs had greater (P = 0.04) jejunal villus height, whereas senior dogs had greater (P < 0.001) colonic crypt depth. Ammonia concentrations decreased (P = 0.03) from proximal to distal colon and were higher in dogs consuming APB (P = 0.03). Age and treatment affected butyrate concentrations, with senior dogs (P = 0.04) and dogs consuming APB (P = 0.04) having higher concentrations. Both diet and age affected small and large intestinal morphology, and colonic fermentative end-product concentrations in dogs. PMID- 16046721 TI - Pullulans and gamma-cyclodextrin affect apparent digestibility and metabolism in healthy adult ileal cannulated dogs. AB - Pullulan and gamma-cyclodextrin are incompletely digestible, glucose-based, nonstructural carbohydrates synthesized by microorganisms. To determine their effect when incorporated into a complete liquid diet on ileal and total tract nutrient digestibility, ileal cannulated dogs (n = 8) were used in a repeated 4 x 4 Latin-square design. Twice daily, diets were offered containing 30% (DMB) maltodextrin, high-molecular-weight (MW) pullulan (MW 100,000), low-MW pullulan (MW 6300), or gamma-cyclodextrin. Fecal and ileal samples were collected for the last 4 d of each 10-d period. Dogs consuming high-MW pullulan had lower (P < 0.05) dry matter, organic matter, crude protein, fat, carbohydrate ileal and total tract digestibilities, and fecal DM, and higher (P < 0.05) fecal output and fecal scores (indicating looser stools). To evaluate glycemic and insulinemic responses to pullulans, food-deprived dogs consumed 25 g maltodextrin, high-MW pullulan, or low-MW pullulan in a repeated 3 x 3 Latin-square design. Glucose and insulin responses were determined for 180 min. Consumption of 25 g alpha-, beta-, and gamma-cyclodextrin resulted in regurgitation within 60 min. High-MW pullulan reduced (P < 0.05) blood glucose concentration at 15, 30, 45, and 60 min. Compared with maltodextrin, low-MW pullulan and gamma-cyclodextrin did not alter nutrient digestibilities or fecal characteristics to any extent, and low MW pullulan did not affect glycemic response. Although high MW pullulan decreased glycemic response, consumption of large amounts negatively affected nutrient digestibility and fecal characteristics. PMID- 16046722 TI - Adaptation to a high-fat diet leads to hyperphagia and diminished sensitivity to cholecystokinin in rats. AB - Rats fed high-fat (HF) diets exhibit reduced sensitivity to some peptide satiety signals. We hypothesized that reduced sensitivity to satiety signals might contribute to overconsumption of a high-energy food after adaptation to HF diets. To test this, we measured daily, 3-h intake of a high-energy, high-fat (HHF, 22.3 kJ/g) test food in rats fed either low-fat (LF) or HF, isoenergetic (16.2 kJ/g) diets. During testing, half of each group received the HHF test food (LF/HHF; HF/HHF), whereas the other half received their respective maintenance diet (LF/LF; HF/HF). Rats fed a HF diet ate more of the HHF food during the 3-h testing period than LF-fed rats (HF/HHF = 7.7 +/- 0.3 g vs. LF/HHF = 5.5 +/- 0.2 g; P = 0.003). Rats tested on their own maintenance diets had similar intakes (HF/HF = 3.2 +/- 0.2 g vs. LF/LF = 3.7 +/- 0.3 g), which were lower (P < or = 0.008) than intakes of rats tested on HHF. HHF-tested rats did not differ in body weight by the end of wk 2 of testing. In a subsequent short-term choice preference test, rats exhibited an equal relative preference for HHF irrespective of their maintenance diets (HF = 63.1%, LF = 68.1%, P = 0.29). Finally, we examined the effect of intraperitoneal NaCl or cholecystokinin (CCK)-8 (100 and 250 ng/kg) injection on 1-h food intake. Both doses of CCK significantly suppressed food intake in LF-fed rats but not HF-fed rats. These results demonstrate that chronic ingestion of a HF diet leads to short-term overconsumption of a high-energy, high-fat food compared with LF-fed cohorts, which is associated with a decreased sensitivity to CCK. PMID- 16046723 TI - Long-chain (n-3) polyunsaturated fatty acids are more efficient than alpha linolenic acid in improving electroretinogram responses of puppies exposed during gestation, lactation, and weaning. AB - Long-chain PUFAs (LCPUFAs) are essential for proper neural and retinal development in many mammalian species. We investigated puppies born to dogs fed diets containing varying amounts of vegetable and marine (n-3) fatty acids during gestation/lactation. The fatty acid compositions of dogs' milk and puppy plasma phospholipids were evaluated, and electroretinographic responses of the young dogs were determined after they were weaned to the same diets. Dogs' milk fatty acid composition reflected the diets fed during gestation/lactation. The milk of dogs fed a high alpha-linolenic acid (ALA) diet was enriched in ALA but not docosahexaenoic acid (DHA). Puppies fed this ALA-enriched milk accumulated more plasma phospholipid DHA than the low (n-3) fatty acid group. However, this accumulation was less than that obtained in puppies fed preformed DHA during development and suckling (P < 0.05). Electroretinograms (ERGs) of 12-wk-old puppies revealed significantly improved visual performance in dogs fed the highest amounts of (n-3) LCPUFAs (P < 0.05). These puppies demonstrated improved rod response (improved amplitude and implicit time of the a-wave, P < 0.05). Puppies from the low (n-3) fatty acid group exhibited the poorest ERG responses compared with the high-marine or high-vegetable (n-3) groups. A novel parameter devised in this study, the initial intensity at which the a-wave was detectable (i.e., threshold intensity), also demonstrated that retinal response of puppies consuming the (n-3) LCPUFA-containing diets occurred at lower light intensity, thereby exhibiting greater rod sensitivity, than the other diet groups. These findings indicate that preformed dietary (n-3) LCPUFA is more effective than ALA in enriching plasma DHA during perinatal development and results in improved visual performance in developing dogs. PMID- 16046724 TI - Dietary fat intake is associated with psychosocial and cognitive functioning of school-aged children in the United States. AB - Using cross-sectional data from the Third National Health and Nutrition Survey, 1988-1994, we examined whether dietary fat intake is associated with cognitive and psychosocial functioning in school-aged children. Based upon 24-h diet recall interviews, dietary intakes of total fat, SFA, monounsaturated fatty, PUFAs, and cholesterol were estimated in 3666 participants aged 6 to 16 y. Psychosocial functioning was evaluated in interviews of each child's mother. Cognitive functioning was measured using achievement and intelligence tests. Overall, total fat and saturated fat were unrelated to measures of cognitive and psychosocial functioning. Compared with equivalent energy intake from saturated fat or carbohydrate, each 5% increase in energy intake from PUFAs was associated with lower risks of poor performance on the digit span test (replacing SFA, OR = 0.58, 95% CI = 0.37-0.91; replacing carbohydrate, OR = 0.61, 95% CI = 0.43-0.88). Cholesterol intake was associated with an increased risk of poor performance on the digit span test (OR = 1.25, 95% CI = 1.11-1.42 for each 100-mg increment intake of cholesterol). The associations were independent of socioeconomic status, maternal education and marital status, and children's nutrition status and were consistent across different methods of energy adjustment in regression models. We conclude that high intake of PUFAs may contribute to an improved performance on the digit span test. In contrast, increased intake of cholesterol may be associated with a poorer performance. PMID- 16046725 TI - Hemoglobin and ferritin are currently the most efficient indicators of population response to iron interventions: an analysis of nine randomized controlled trials. AB - Governments and donor agencies have implemented pilot and large-scale iron fortification programs, but there has been no consensus on the best choice of indicators to monitor population response to these interventions. We analyzed data from 9 randomized iron intervention trials to determine which of the following indicator(s) of iron status show the largest response in a population: hemoglobin (Hb), ferritin, transferrin receptor (TfR), zinc protoporphyrin (ZPP), mean cell volume (MCV), transferrin saturation (TS), and total body-iron store. We expressed the change in each indicator in response to the iron intervention in SD units (SDU) for the intervention group compared with the control group. Ferritin increased by > or =0.2 SDU in all trials and was significant in 7. Hb changed by > or =0.2 SDU in 6 and was significant in 5. TfR increased by > or =0.2 SDU in 5 of 8 interventions in which it was measured and was significant in 4. ZPP increased by > or =0.2 SDU and was significant in 3 of 6 interventions. Excluding Hb, the indicator with the largest change in SDU was ferritin in 4 trials, TS in 2 trials, body-iron store in 2 trials, and TfR in 1. In the 2 cases in which body-iron stores showed the largest change, the change in ferritin was nearly as large. Our results suggest that with currently available technologies, ferritin shows larger and more consistent response to iron interventions than ZPP or TfR. We cannot make confident inference about MCV or TS, which were included in only 4 and 2 trials, respectively. It is possible that the optimal indicator(s) may differ with age, sex, and pregnancy. There were too few trials in each age and sex group to allow us to explore this question. PMID- 16046726 TI - A homocysteine metabolism-related dietary pattern and the risk of coronary heart disease in two independent German study populations. AB - A biomarker profile of high folate and vitamin B-12 and low plasma homocysteine concentrations reduces the risk of coronary heart disease (CHD) and may be linked to diet. The objectives of the present study were to identify a food pattern related to these biomarkers and to examine its association with CHD risk. Dietary patterns related to biomarker plasma concentrations were constructed from data obtained in the Coronary Risk Factors for Atherosclerosis in Women (CORA) Study (200 cases; 255 controls) using the reduced rank regression statistical method. Risks for CHD with relation to the identified pattern were estimated in the CORA study and in the European Prospective Investigation into Cancer and Nutrition (EPIC)-Potsdam Study with 157 cases of incident myocardial infarction among 26,795 participants. In these 2 German study populations, whole-grain bread, fresh fruit, olive oil, mushrooms, cruciferous vegetables, wine, and nuts contributed the most positively and fried potatoes the most negatively to a dietary pattern that was directly associated with both plasma folate and vitamin B-12 concentrations, but inversely with plasma homocysteine. Multivariate adjusted relative risks for CHD across increasing quintiles of the food pattern score were 1.0, 0.55, 0.52, 0.58, 0.39 (P for trend = 0.05) in the case-control sample and 1.0, 0.95, 0.75, 0.56, 0.72 (P for trend = 0.041) in the prospective study. The combination of a high intake of whole-grain bread, fresh fruit, olive oil, mushrooms, cruciferous vegetables, wine, and nuts with a low intake of fried potatoes was associated with a favorable biomarker profile of homocysteine metabolism and reduced risk of CHD. PMID- 16046727 TI - Polymorphisms in cytoplasmic serine hydroxymethyltransferase and methylenetetrahydrofolate reductase affect the risk of cardiovascular disease in men. AB - Genetic variation in folate-regulating enzymes contributes to the risk of cardiovascular disease (CVD). The cytoplasmic serine hydroxymethyltransferase (cSHMT) enzyme is proposed to regulate a key metabolic intersection in folate metabolism. We hypothesized that a variant in cSHMT (cSHMT 1420C-->T) affects CVD risk, and that the effect depends on a linked step in the metabolic pathway catalyzed by methylenetetrahydrofolate reductase (MTHFR). A nested case-control study of incident CVD was conducted within the all-male Normative Aging Study cohort. Of the incident CVD cases, 507 had DNA samples; 2 controls/case were selected by risk set sampling (matched on age and birth year). A significant gene gene interaction (P-values 0.0013, 0.0064) was found between MTHFR and cSHMT, and there was little or no change in the coefficients in covariate-adjusted models. The effect of MTHFR 677C-->T genotype on CVD risk varied by cSHMT 1420C-->T genotype. Among men with cSHMT 1420C-->T TT genotype, the odds ratios (OR) for CVD risk for MTHFR 677C-->T CT and TT genotypes compared with the MTHFR 677C-->T CC genotype were 3.6 (95% CI, 1.7-7.8) and 10.6 (95% CI, 2.5-46.0), respectively. Among men with the cSHMT 1420C-->T CC/CT genotype, the corresponding ORs were 1.0 (95% CI, 0.8-1.2) and 1.3 (95% CI, 0.9-1.8). Plasma total homocysteine concentrations were highest in the subgroup of men with both polymorphisms, MTHFR 677C-->T TT and cSHMT 1420C-->T TT, consistent with a higher risk of CVD in this subgroup. A more complete understanding of the molecular mechanism awaits identification of the functional effect of the polymorphism. PMID- 16046728 TI - Natural chlorophyll but not chlorophyllin prevents heme-induced cytotoxic and hyperproliferative effects in rat colon. AB - Diets high in red meat and low in green vegetables are associated with an increased risk of colon cancer. In rats, dietary heme, mimicking red meat, increases colonic cytotoxicity and proliferation of the colonocytes, whereas addition of chlorophyll from green vegetables inhibits these heme-induced effects. Chlorophyllin is a water-soluble hydrolysis product of chlorophyll that inhibits the toxicity of many planar aromatic compounds. The present study investigated whether chlorophyllins could inhibit the heme-induced luminal cytotoxicity and colonic hyperproliferation as natural chlorophyll does. Rats were fed a purified control diet, the control diet supplemented with heme, or a heme diet with 1.2 mmol/kg diet of chlorophyllin, copper chlorophyllin, or natural chlorophyll for 14 d (n = 8/group). The cytotoxicity of fecal water was determined with an erythrocyte bioassay and colonic epithelial cell proliferation was quantified in vivo by [methyl-(3)H]thymidine incorporation into newly synthesized DNA. Exfoliation of colonocytes was measured as the amount of rat DNA in feces using quantitative PCR analysis. Heme caused a >50-fold increase in the cytotoxicity of the fecal water, a nearly 100% increase in proliferation, and almost total inhibition of exfoliation of the colonocytes. Furthermore, the addition of heme increased TBARS in fecal water. Chlorophyll, but not the chlorophyllins, completely prevented these heme-induced effects. In conclusion, inhibition of the heme-induced colonic cytotoxicity and epithelial cell turnover is specific for natural chlorophyll and cannot be mimicked by water-soluble chlorophyllins. PMID- 16046729 TI - Direct detection and evaluation of conversion of D-methionine into L-methionine in rats by stable isotope methodology. AB - The stereoselective kinetics of methionine enantiomers in rats was investigated to evaluate the fraction that converted from d-methionine to the L-enantiomer using a stable isotope methodology. After bolus i.v. administration of D- or L [(2)H(3)]methionine, their plasma concentrations and that of endogenous L methionine were determined by a stereoselective GC-MS method. L [(2)H(3)]Methionine appeared rapidly after administration of D [(2)H(3)]methionine, whereas D-[(2)H(3)]methionine was not detected after administration of L-[(2)H(3)]methionine. The fraction of conversion of D [(2)H(3)]methionine into L-[(2)H(3)]methionine was estimated using the area under the plasma concentration vs. time curve of L-[(2)H(3)]methionine on D [(2)H(3)]methionine administration and total clearance of L-[(2)H(3)]methionine on L-[(2)H(3)]methionine administration, and that fraction was >90%. This result demonstrates that almost all i.v. administered D-methionine is converted into the L-enantiomer in vivo. PMID- 16046730 TI - Challenges and opportunities for support of nutritional science conferences and scientific meetings at National Institutes of Health. AB - Opportunities exist throughout the institutes, centers, and offices of the NIH for obtaining support for conferences and scientific meetings. The number of conferences funded by NIH that thematically involve nutrition, health, and biomedical research is very small relative to the total number of conference grants awarded by NIH. Only 4% of conference grant (R13 and U13) applications awarded by all institutes and centers of the NIH over the last 5 y were determined to be focused on nutrition in some way. The National Cancer Institute has the largest proportion of these awards, and the numbers of such applications submitted each year appear to be increasing. Only approximately 18 nutrition research-related conference grant applications appear to have been funded, on average, per year over the last 5 y. Success rates are high; an average of 85% of nutrition-related applications submitted over the last 5 y were funded. The purpose of this article is to encourage applications from the nutrition community and to provide information for consideration when developing an application for a conference or workshop. PMID- 16046731 TI - Executive summary report. PMID- 16046732 TI - Tomato products, lycopene, and prostate cancer: a review of the epidemiological literature. PMID- 16046733 TI - Variations in lycopene blood levels and tomato consumption across European countries based on the European Prospective Investigation into Cancer and Nutrition (EPIC) study. PMID- 16046734 TI - Directions for future epidemiological research in lycopene and prostate cancer risk. PMID- 16046735 TI - How can the metabolomic response to lycopene (exposures, durations, intracellular concentrations) in humans be adequately evaluated? PMID- 16046736 TI - What are typical lycopene intakes? PMID- 16046737 TI - How do nutritional and hormonal status modify the bioavailability, uptake, and distribution of different isomers of lycopene? PMID- 16046738 TI - What can pharmacokinetic models tell us about the disposition of lycopene and the potential role of lycopene in cancer prevention? PMID- 16046739 TI - Lycopene effects on rat normal prostate and prostate tumor tissue. PMID- 16046740 TI - Can smoke-exposed ferrets be utilized to unravel the mechanisms of action of lycopene? PMID- 16046741 TI - Tomatoes or lycopene: a role in prostate carcinogenesis? PMID- 16046742 TI - Are there adverse effects of lycopene exposure? PMID- 16046743 TI - How do intermediate endpoint markers respond to lycopene in men with prostate cancer or benign prostate hyperplasia? PMID- 16046744 TI - Intermediate biomarkers of lycopene/tomato effects in high-risk prostatic tissue. PMID- 16046745 TI - Selection of surrogate endpoint biomarkers to evaluate the efficacy of lycopene/tomatoes for the prevention/progression of prostate cancer. PMID- 16046746 TI - How strong is the evidence that lycopene supplementation can modify biomarkers of oxidative damage and DNA repair in human lymphocytes? PMID- 16046747 TI - A research agenda for lycopene/tomato supplementation and cancer prevention. PMID- 16046749 TI - Incidental radiological diagnosis of rickets. AB - Rickets fortunately remains rare in the United Kingdom, although its actual incidence is currently undetermined.1 Many still consider it to be a disease of poverty prevalent during the Victorian era. However, a number of recent articles have highlighted concern among British health professionals about the number of cases still being diagnosed in this country. These cases have nearly all involved non-Caucasian children who are considered to be at high risk due to skin colour, prolonged breast feeding, and low maternal vitamin D levels. Their presentations are variable ranging from failure to thrive, bone deformities, seizures, and even stridor. The diagnosis is usually made in babies and toddlers.We present a series of patients attending our accident and emergency (A&E) department, over a five month period, where the diagnosis of rickets was primarily a radiological diagnosis. PMID- 16046750 TI - Cranial computed tomography in trauma: the accuracy of interpretation by staff in the emergency department. AB - OBJECTIVES: Cranial computed tomography (CT) is replacing skull radiography in head trauma. Rapid radiological opinions on these images may not always be available. We assessed the ability of our permanent emergency department staff to interpret the images. METHODS: A retrospective series of 100 consecutive cases was reviewed and interpreted by five permanent emergency department medical staff, and their interpretation compared with the consensus opinion of two radiologists. RESULTS: An overall agreement of 86.6% (95% confidence interval (CI) 83.4 to 89.9) was achieved, with a false negative rate of 4.2% (95% CI 3.9 to 4.3). No findings that would have changed the overnight management of any patient were missed. CONCLUSIONS: Our results for CT scans are similar to studies of interpretation of other radiographic images in emergency departments. Our emergency staff could safely make the initial interpretation of cranial CT images in trauma out of hours, and formal reporting may wait until a suitably experienced radiologist is available. PMID- 16046751 TI - CT or not CT--that is the question. Whether 'tis better to evaluate clinically and x ray than to undertake a CT head scan! AB - OBJECTIVE: To evaluate the usage of computed tomography (CT) head scanning in children at the Royal Aberdeen Children's Hospital after the publication of the National Institute of Clinical Excellence (NICE) guidelines on the management of head injury. METHODS: The Accident and Emergency case records of all children presenting with a head injury over a three month period were reviewed and the number of attendances, radiographs, and CT head scans undertaken were noted. Also noted was the number of additional CT head scans that would have been performed if the NICE guidelines had been rigidly followed. RESULTS: Five hundred and thirty seven children were included in the study: 67% were boys. Two hundred and ten (39%) had skull radiographs: six demonstrated skull fractures and eight (1.5%) underwent CT head scan, with one positive report of a skull fracture. There were no reports of intracranial abnormalities. Ninety nine (18.4%) were admitted. Strictly applying all the NICE criteria for CT scanning would have resulted in an additional 54 patients being scanned. CONCLUSION: Rigid adherence to the NICE guidelines in all children with head injuries would have resulted in an almost eightfold increase in CT head scans performed. None of these children had clinical signs of intracranial injury and would have been exposed to a large amount of ionising radiation. The use of guidelines in practice must always be considered in conjunction with clinical judgement. PMID- 16046752 TI - The use of a template to improve the management of distal radial fractures. AB - OBJECTIVES: The aim of this study was to reduce the number of inappropriately managed distal radial fractures with the use of a template. METHODS: A template has been developed to aid junior doctors and emergency nurse practitioners (ENPs) to decide which distal radial fractures require manipulation. Emergency Department (ED) junior doctors and ENPs were asked to review the radiographs of 12 distal radial fractures with and without the template and comment on whether the fracture required manipulation. RESULTS: There was an improvement in appropriate management with the template of 16.6% for the junior doctors and 22.3% for the ENPs. This was statistically significant for both groups when the results were analysed with a paired t test (p<0.01). CONCLUSIONS: We conclude that the availability of this template in the ED may be helpful to junior doctors and ENPs, and represents a low cost and efficient way of reducing the number of patients who attend a fracture clinic with a distal radial fracture in an unsatisfactory position. This may also reduce the numbers requiring admission and a manipulation under anaesthetic, which spares the patients the risks of general anaesthesia and may possibly be cost saving. PMID- 16046753 TI - "See and Treat": spreading like wildfire? A qualitative study into factors affecting its introduction and spread. AB - OBJECTIVES: The aim of this paper was to explore key factors that influenced the spread of "See and Treat" in a range of accident and emergency (A&E) departments. METHODS: The study adopted a qualitative approach, and semi-structured interviews were undertaken with 21 key individuals working across 10 A&E departments operating See and Treat. Participants included clinicians, managers, and chief executives. RESULTS: Many factors influenced the spread of See and Treat. The initiative was well supported and monitored by external agencies, patients benefited and no staff groups lost out, waiting times were reduced, and Department of Health targets were achieved. However, this study indicates there were also a range of factors that limited the spread of See and Treat, including lack of additional resources and suitably experienced staff, impact upon quality of care, and no prior evaluation of its benefits. An interesting additional factor that may be both facilitating and limiting is the complexity of the A&E culture, in particular staff perspectives about working with minor injuries. CONCLUSIONS: See and Treat was promoted as a solution to waiting times problems in A&E, without evidence from any national evaluation. However, many staff members referred to its usefulness as a tool to reduce waiting times and enhance the patient journey, although resource, quality, and staffing issues may mean such an initiative may be difficult to sustain in its present form. PMID- 16046754 TI - Survey of attitudes of senior emergency physicians towards the introduction of emergency department ultrasound. AB - INTRODUCTION: Emergency department ultrasound (EDU) is widely practised in the USA, Australia, parts of Europe, and Asia. EDU has been used in the UK since the late 1990s but as yet, few areas have established a practice. OBJECTIVES: To assess the current climate of opinion with respect to the practice, constraints, and establishment of EDU among emergency department (ED) consultants on the island of Ireland. METHODS: A postal questionnaire was formulated, piloted, and assessed for ambiguity by a sample of ED consultants and an independent non-ED consultant, prior to being mailed to all ED consultants in Ireland. RESULTS: Of the 58 consultants canvassed 46 (79%) responded. Of the respondents, 40 (87%) strongly agreed/agreed that EDU is appropriate and should be performed in the ED. Of these, 3 (7%) are currently performing EDU; 37 (80%) have not had formal training in EDU, however 42 (91%) support the establishment of national guidelines for training in focused ultrasound in the ED. Problems instituting EDU were often multifactorial. Commonly highlighted difficulties included financial issues (24 respondents, 52%) and radiology department support (16 respondents, 34%). Other cited problems include varying interdepartmental practices (15 respondents, 33%) and (for some EDs) low numbers of patients requiring EDU, with projected difficulties in skills maintenance. CONCLUSION: Despite the vast majority of ED consultants being in favour of EDU, very few actually perform it on a regular basis or have had any formal training. Highlighted difficulties in EDU implementation included financial constraints, lack of support from radiology departments, and lack of formal training. PMID- 16046755 TI - New insights into the glucose oxidase stick test for cerebrospinal fluid rhinorrhoea. AB - Rhinorrhoea is a clinical sign of cerebrospinal fluid (CSF) leakage in patients with skull fracture, but can also be attributable to respiratory secretions or tears. Laboratory tests confirming the presence of CSF are not sufficiently rapid to support clinical decision making in the emergency department and may not be universally available. Detection of glucose in nasal discharge was traditionally used to diagnose CSF leak at the bedside, but has fallen into disuse as it has poor positive predictive value. We propose an algorithm to improve the diagnostic value of this test taking into consideration factors we have found to affect the glucose concentration of respiratory secretions. In patients at risk of CSF leak, nasal discharge is likely to contain CSF if glucose is present in the absence of visible blood, if blood glucose is <6 mmol x L(-1), and if there are no symptoms of upper respiratory tract infection. PMID- 16046757 TI - Best evidence topic report. Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. PMID- 16046756 TI - Best evidence topic report. Fasting before prilocaine Biers' block. AB - A short cut review was carried out to establish whether a period of fasting increases the safety of Biers' block (intravenous regional anaesthesia). A total of 50 papers were found using the reported search, of which four presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 16046758 TI - Best evidence topic report. Ultrasound in the diagnosis of testicular torsion. AB - A short cut review was carried out to establish whether colour Doppler ultrasound is more sensitive than clinical examination in ruling out testicular torsion. A total of 284 papers were found using the reported search, of which seven presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 16046759 TI - Best evidence topic report. Primary split skin grafts for pretibial lacerations. AB - A short cut review was carried out to establish whether primary split skin grafting is better than simple wound edge approximation at reducing time to healing in patients with pretibial flap lacerations. A total of 72 papers were found using the reported search, of which one presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of this best paper are tabulated. A clinical bottom line is stated. PMID- 16046760 TI - Best evidence topic report. Serial x rays in battery ingestion. AB - A short cut review was carried out to establish whether serial x rays were necessary in cases of battery ingestion where the battery has passed through the oesophagus. A total of 104 papers were found using the reported search, of which none presented any evidence to answer the clinical question. It is concluded that there is no evidence available to answer this question. Further research is needed. PMID- 16046761 TI - SOCRATES 10 (synopsis of Cochrane reviews applicable to emergency services). PMID- 16046762 TI - 15 assessment and management of neurological problems (2). PMID- 16046763 TI - A review of emergency equipment carried and procedures performed by UK front line paramedics on paediatric patients. AB - OBJECTIVES: In 1997 a review of paramedic practice upon adult patients in the UK found many inconsistencies and deficiencies in basic care. A follow up review in 2002 identified widespread improvement in provision of equipment and skills to provide basic and advanced life support.Paediatric care was not assessed in either review. The authors conducted this study to identify current standards of care in paediatric paramedic practice and areas of potential improvement. METHOD: A questionnaire designed to determine what equipment and skills were available to paramedics for the management of common or serious paediatric emergencies was sent to chief executives of the 32 NHS Ambulance Trusts in England and Wales. RESULTS: The trend of expanding and standardising practice among adult patients has not extended to paediatric practice despite national guidelines from the Joint Royal Colleges Ambulance Liaison Committee (JRCALC). Furthermore there are some serious failings in the provision of care and skills. Many Trusts have not adopted JRCALC guidelines for the management of life threatening paediatric emergencies such as asthma, meningitis, and fluid replacement in hypovolaemia. CONCLUSIONS: Ambulance Trusts not meeting standards set out in the JRCALC guidelines must address their areas of deficiency. Failure to do so endangers children's lives and leaves Trusts open to criticism. PMID- 16046764 TI - Case based learning--a review of the literature: is there scope for this educational paradigm in prehospital education? AB - This paper discusses the findings of a literature review of case based learning (CBL) from a multidisciplinary health science education perspective and attempts to draw comparisons with the available literature relating to prehospital education and CBL.CBL is an exciting educational prospect in which to develop research capacity, strategies, and opportunities. This paper provides an examination of the literature exploring the major consistencies and inconsistencies, and reveals areas of potential future research for prehospital education institutions. PMID- 16046765 TI - Retrospective observational case-control study comparing prehospital thrombolytic therapy for ST-elevation myocardial infarction with in-hospital thrombolytic therapy for patients from same area. AB - OBJECTIVES: To compare a system of prehospital thrombolytic therapy, delivered by paramedics under medical guidance, with in-hospital thrombolytic therapy in meeting National Service Framework (NSF) targets for treatment of acute myocardial infarction at a District General Hospital setting in England. DESIGN: Retrospective observational case-control study comparing patients with suspected acute myocardial infarction (AMI) treated with thrombolytic therapy in the prehospital environment with patients treated in hospital. SETTING: Wyre Forest District and Worcestershire Royal Hospital, UK. PARTICIPANTS: (A) All patients who received prehospital thrombolytic therapy for suspected AMI accompanied by electrocardiographic features considered diagnostic.(B) Patients who received thrombolytic therapy after arrival at hospital for the same indication, matched with group A by age, gender and postcode. MAIN OUTCOME MEASURES: 1. Call to needle time. 2. Percentage of patients treated within one hour of calling for medical help. 3. Appropriateness of thrombolytic therapy. 4. Safety of thrombolytic therapy RESULTS: 1. The median call to needle time for patients treated before arriving in hospital (n = 27) was 40 minutes with an inter quartile range 25-112 (mean 43 minutes). Patients from the same area who were treated in hospital (n = 27) had a median time of 106 minutes with an inter quartile range 50-285 (mean 126 minutes). This represents a median time saved by prehospital treatment of 66 minutes. 2. 60 minutes after medical contact, 96 % of patients treated before arrival in hospital had received thrombolytic therapy; this compares with 4% of patients from similar areas treated in hospital. 3. Myocardial infarction was confirmed in 92% (25/27) of patients who received prehospital thrombolytic therapy and similarly 92% (25/27) of those given in hospital thrombolytic therapy. 4. No major bleeding occurred in either group. Group A suffered fewer in-hospital deaths than group B (1 versus 4). Cardiogenic shock (3 patients) and ventricular arrhythmia (5 patients) were seen only in group B. CONCLUSION: Paramedic-delivered thrombolytic therapy can be delivered appropriately, safely, and effectively. Time gains are substantial and can meet the national targets for early thrombolytic therapy in the majority of patients. PMID- 16046766 TI - Superior sagittal sinus thrombosis, an unusual presentation of acute myeloid leukaemia: a case report. PMID- 16046767 TI - A young man with a headache. PMID- 16046768 TI - An unusual cause of an acutely locked knee. PMID- 16046769 TI - Myocardial hamartoma as a cause of VF cardiac arrest in an infant. PMID- 16046770 TI - A case report of an unusual sternal fracture. AB - Most sternal fractures are transverse, and a lateral chest radiograph is diagnostic. We report a case of vertical sternal fracture that was not seen on plain radiographs but was revealed using computed tomography (CT). Thoracic CT with coronal reformatted images can also demonstrate sternal fracture lines, supernumerary synchondrosis, and costosternal joint abnormalities. PMID- 16046771 TI - Eureka! A surprising appearance after aspiration of a haemopneumothorax: treat the patient not the radiograph. AB - We describe the aspiration of a traumatic haemopneumothorax and an unexpected anteroposterior chest radiograph finding after the procedure. Chest aspiration is now routine emergency management for spontaneous pneumothorax. There have been no previous documented reports of this clinical scenario of radiological deterioration with clinical improvement after aspiration of a haemopneumothorax. PMID- 16046772 TI - Renal infarction mimicking renal colic in patient with a prosthetic aortic valve. AB - A 31 year old man with prosthetic aortic valve replacement presented with sudden onset of colic right flank pain. Analysis of the urine revealed haematuria, and the international normalised ratio was suboptimal. The patient was misdiagnosed as having ureteral colic. On the second day, an ultrasound showed no signs of obstructive uropathy, and there was no evidence of absent function on intravenous pyelogram. Computed tomography with contrast agent was performed and revealed a right renal infarction. Renal angiography demonstrated total occlusion of the right renal artery. Fibrinolytic therapy and angioplasty were unsuccessful. To our knowledge, aortic prosthetic valve thrombus as a source of renal artery embolism mimicking renal colic has not been reported previously. This case underlines the importance of renal colic as a manifestation of renal infarction in patients with prosthetic valves and the need for a high index of suspicion of renal embolism. PMID- 16046773 TI - The problem with nose bleeds. AB - Epistaxis is common in the paediatric population and is usually minor and self limiting. This case illustrates an atypical presentation of epistaxis with hypovolaemic shock due to a dissecting false aneurysm of the internal carotid artery caused by an impalement injury to the oropharynx. PMID- 16046774 TI - Tension pneumothorax and the "forbidden CXR". AB - A case is presented of unilateral tension pneumothorax associated with flail chest and pulmonary contusions in a spontaneously ventilating patient after a fall. The tension element was not suspected until chest x ray was available, nor was immediate needle thoracocentesis performed. No morbidity resulted as a consequence. This case highlights the difficulty in deciding whether or not tension pneumothorax is the predominant cause of respiratory distress in a patient with multiple chest injuries. It provides further evidence challenging some of the doctrine on how to treat suspected tension pneumothorax. PMID- 16046775 TI - Troponin for prediction of cardiovascular collapse in acute colchicine overdose. AB - The use of colchicine, a treatment for acute gout and familial Mediterranean fever, is limited by its toxicity. A relatively low dose of colchicine may be fatal. After a colchicine overdose, monitoring should include 6-12 hourly serum troponin measurements. A rising troponin level predicts cardiovascular collapse and is an indication for more intensive management. PMID- 16046776 TI - Long QT syndrome presenting as epileptic seizures in an adult. AB - A 50 year old woman with a previous diagnosis of epilepsy presented to the emergency department with a generalised seizure. Her admission ECG showed QT prolongation secondary to bradycardia and a subsequent seizure in the department demonstrated that these events were secondary to cerebral hypoperfusion during episodes of torsades de pointes. This case illustrates how long QT syndrome can masquerade convincingly as epilepsy, delaying treatment and exposing the patient to a high risk of sudden cardiac death. Careful ECG analysis is recommended for all patients presenting with seizures. PMID- 16046777 TI - C-reactive protein: a valuable acute investigation. A case of pneumococcal meningitis presenting as ankle pain. AB - A case is presented in which the decision to admit and treat an adult with musculoskeletal pain and pyrexia was based on her markedly raised c-reactive protein (CRP). At the time of admission she was apyrexial and the CRP was the only haematological investigation that was out of the normal range. She subsequently became precipitously septic with pneumococcal bacteraemia and meningitis. The CRP is an important investigation for emergency departments. PMID- 16046778 TI - Misdiagnosis of myocardial infarction by troponin I following minor blunt chest trauma. PMID- 16046779 TI - Brugada syndrome: syncope in the younger patient and the risk of sudden cardiac death. PMID- 16046780 TI - Spinal epidural haematoma mimicking spontaneous subarachnoid haemorrhage. AB - Spinal epidural haematoma is increasingly picked up using magnetic resonance imaging, especially following trauma. The presentation can be varied especially if there is a trivial history of trauma. Spontaneous subarachnoid haemorrhage can occur in spinal haematoma as a rare and unusual presentation and can be easily misdiagnosed. The key to early diagnosis is a strong suspicion and careful repeated neurological examination. We present an illustrative case. PMID- 16046781 TI - Ultrasound guidance for central venous catheter insertion. PMID- 16046782 TI - Training in anaesthesia is also an issue for nurses. PMID- 16046783 TI - Anaesthetic eye drops for children in casualty departments across south east England. PMID- 16046784 TI - Unnecessary Tetanus boosters in the ED. PMID- 16046785 TI - Patients' attitudes toward medical photography in the emergency department. PMID- 16046786 TI - Environmental cardiovascular disease. AB - The last decade has seen a remarkable growth in the evidence establishing exposure to environmental agents as a risk factor for cardiovascular disease (CVD). Most of this has come from research linking exposure to ambient particulate matter with CVD, although more recent evidence suggests that the ozone might also be contributing factor. Research on the cardiovascular toxicity of other pollutants, notably arsenic, has also grown during this period. In addition to their effects in adults, environmental agents, such as dioxin, have also been shown to adversely affect development of the heart in laboratory animals. Taken together, these results suggest that environmental exposure must be considered as an important risk factor for CVD and that further research to determine mechanisms of action and susceptible populations is needed. This research will require interdisciplinary approaches, and should yield data not only on the toxic effects of pollutants on the cardiovascular system, but on the basic pathophysiology of cardiovascular diseases as well. PMID- 16046787 TI - The transcribed genome and the heritable basis of essential hypertension. AB - Gene expression can be now assessed quantitatively and comprehensively. In addition to reflecting the specialized differentiation of the cell or tissue type from which gene expression is sampled, it also manifests patterns determined by inheritance. Thus gene expression is a phenotypic trait, at least when assessed comprehensively. This trait shows familial aggregation and segregation patterns indicative of an inherited contribution. The molecular evolution of genes includes mutations affecting regulatory sequences in the genome that influence gene expression in cis and in trans. Such mutations may increase in frequency in a population either by genetic drift or by selection. Traits of gene expression, acting alone or in concert with other gene expression traits, may generate phenotypes that extend beyond transcript abundance. Indeed, the divergence of species and the traits that distinguish related species appear to rely importantly on inherited divergence in the control of gene expression. Variation in gene expression may contribute to the pathogenesis of a prevalent human disease trait that shows heritability--essential hypertension. Along with other common heritable diseases, hypertension susceptibility arises from the actions of multiple genome sequence variations. The identity of such variation has proven elusive when sought by methods that have been successfully applied to Mendelian diseases. This review explores the potential to uncover hypertension genes by exploiting quantitative variation in the heritable control of gene expression. PMID- 16046788 TI - A gene-environment interaction model of stress-induced hypertension. AB - The case for a gene-environment interaction model of stress-induced hypertension is detailed in this paper. We hypothesize that repeated exposure to stress in combination with an environmentally and/or genetically mediated susceptibility may lead to the development of essential hypertension. Previously, we reviewed the evidence for a genetic influence on the two major intermediate phenotypes of our model: cardiovascular reactivity to psychological stress and stress-induced sodium retention, representing the cardiovascular and renal stress response, respectively. Here we first describe how genes underlying the physiological systems mediating the stress response of heart, vasculature, and kidney (i.e., the sympathetic nervous system, renin-angiotensin- aldosterone system and sodium reabsorption, and the endothelial system) may increase vulnerability to stress and confer susceptibility to development of essential hypertension. Next, we extend our model and review genes underlying three additional systems that may mediate the influence of stress on the development of essential hypertension: the parasympathetic nervous system, the serotonergic system, and the hypothamamus pituitary-adrenal axis. The elucidation of our gene-environment interaction model of stress-induced essential hypertension will improve the understanding of the contribution of stress to the development of essential hypertension. This knowledge may lead to more effective primary and secondary prevention programs involving lifestyle interventions in which the role of stress, both acute and chronic, will be taken into account, particularly for individuals at increased genetic risk of essential hypertension. PMID- 16046789 TI - Hypertension and SNP genotyping in antihypertensive treatment. AB - Hypertension is prevalent, affecting approx 20--25% of the adult population in the Western world. Primary hypertension is a multifactorial, complex disorder where many genes and genetic variants are assumed to interact with environmental factors in order to produce the specific blood pressure level for a given individual. Family and twin studies show that between 30 and 60% of blood pressure variation is determined by genetic factors. Monogenic disorders of hypertension are rare and do not explain blood pressure variability in the population at large. Obvious candidate genes for the study of hypertension are those that encode components of a blood pressure regulating system targeted by an antihypertensive drug, or those that are involved in counter-regulatory systems. In this review, we give a brief pathophysiological background to hypertension and the rational behind utilizing SNP genotyping in the study of hypertension and the antihypertensive response to treatment. We also discuss some of the novel results of pharmacodynamic studies in antihypertensive treatment, an area in its infancy. PMID- 16046790 TI - Atherosclerosis: from genetic polymorphisms to system genetics. AB - It is generally considered that the genetics of atherosclerosis and its complications involves a large number of genes with common alleles having weak effects on disease risk but possibly interacting with each other and with non genetic factors. In such a complex system, absence of marginal effects (effects of polymorphisms considered one at a time) is insufficient to exclude the implication of a polymorphism on disease risk. Investigating polymorphisms and even genes one by one is no longer appropriate. It is necessary to focus on biological systems and integrate the contribution of genetic as well as non genetic factors and their interactions. Hopefully, system genetics will ultimately improve our understanding of the genetic architecture of complex traits. PMID- 16046791 TI - Modification of environmental toxicity by nutrients: implications in atherosclerosis. AB - We hypothesize that nutrition can modulate the toxicity of environmental pollutants and thus modulate health and disease outcome associated with chemical insult. There is now increasing evidence that exposure to persistent organic pollutants, such as PCBs, can contribute to the development of inflammatory diseases such as atherosclerosis. Activation, chronic inflammation, and dysfunction of the vascular endothelium are critical events in the initiation and acceleration of atherosclerotic lesion formation. Our studies indicate that an increase in cellular oxidative stress and an imbalance in antioxidant status are critical events in PCB-mediated induction of inflammatory genes and endothelial cell dysfunction. Furthermore, we have found that specific dietary fats can further compromise endothelial dysfunction induced by selected PCBs and that antioxidant nutrients (such as vitamin E and dietary flavonoids) can protect against endothelial cell damage mediated by these persistent organic pollutants. Our recent data suggest that membrane lipid rafts such as caveolae may play a major role in the regulation of PCB-induced inflammatory signaling in endothelial cells. In addition, PCB- and lipid-induced inflammation can be down-regulated by ligands of anti-atherogenic peroxisome proliferator-activated receptors (PPARs). We hypothesize that PCBs contribute to an endothelial inflammatory response in part by down-regulating PPAR signaling. Our data so far support our hypothesis that antioxidant nutrients and related bioactive compounds common in fruits and vegetables protect against environmental toxic insult to the vascular endothelium by down-regulation of signaling pathways involved in inflammatory responses and atherosclerosis. Even though the concept that nutrition may modify or ameliorate the toxicity of environmental chemicals is provocative and warrants further study, the implications for human health could be significant. More research is needed to understand observed interactions of PCB toxicity with nutritional interventions. PMID- 16046792 TI - The role of phospholipases in lipid modification and atherosclerosis. AB - Phospholipases have received wide attention as it has become clear that several isoforms of the phospholipase family play a role in onset and progression of atherosclerosis. The release of free fatty acids (FFA) and lysophospholipids (lysoPL) provide metabolites for various inflammatory pathways, and this has been considered the main mechanism of phospholipase-driven inflammation. However, generation of FFA and lysoPL are only part of the story. The induction of low density phospholipoprotein (LDL) aggregation and accumulation, receptor binding, co-regulation with cyclooxygenase (COX) and lip-oxygenase (LO) pathways, internalization through heparan sulfate proteoglycan (HSPG) shuttling, and crosstalk between phospholipases all play a role in atherosclerosis.Group IIA phospholipase has long been considered a key enzyme in the initiation of various inflammatory diseases, but new data also indicate a role in the subsequent resolution of inflammatory processes. Recently, secreted group V and group X phospholipase and platelet activating factor acetylhydrolase (PAF-AH) are also recognized as important enzymes in atherosclerosis, modifying LDL and leading to lipid accumulation. The phospholipases and their function in atherosclerosis are not fully under-stood. Future investigations can deliver better insight in the complex role of these enzymes. The present review summarizes the current state of phospholipase research related to atherosclerosis. PMID- 16046793 TI - NF-kappaB in cardiovascular disease: diverse and specific effects of a "general" transcription factor? AB - The transcription factor NF-kappaB regulates a wide variety of biological effects in diverse cell types and organs, particularly stress and adaptive responses. Recently, it has become recognized that NF-kappaB and its upstream regulator tumor necrosis factor (TNF)-alpha regulate specific antithetical effects. For instance, in the heart, NF-kappaB has been found to be required for development of late preconditioning against myocardial infarction and yet is critically involved in mediating cell death after ischemia/reperfusion injury. There remains a bias that NF-kappaB is a "general" transcription factor that is activated by a plethora of stimuli, including neurohormonal, pathophysiological, and stress stimuli, and affects regulation of numerous downstream genes. The question has become, how can such a "general" transcription factor be critically involved in mediating specific effects? An emerging hypothesis is that NF-kappaB is part of a complicated signaling network or web, and that different combinatorial interactions between various activated signaling pathway components produce specific outcomes. This idea is supported by the large number of interactions discovered in the past 14 years between NF-kappaB and other signaling pathways at multiple levels. Notwithstanding the complexities of signal-induced activation of NF-kappaB, since it is a transcription factor, specific effects of NF-kappaB activation must be underlain by the activation and/or suppression of distinct subsets of NF-kappaB-dependent genes. At this level, selectivity is conferred by the expression of specific NF-kappaB subunits, their post translational modifications, and by combinatorial interactions between NF-kappaB and other transcription factors and coactivators that form specific enhanceosome complexes in association with particular promoters. These enhanceosome complexes represent another level of signaling integration whereby the activities of multiple upstream pathways converge to impress a distinct pattern of gene expression upon the NF-kappaB-dependent transcriptional network. Understanding how the overall cellular signaling network translates NF-kappaB activation into the regulation of specific subsets of NF-kappaB-dependent genes will lead to a mechanistic understanding of how NF-kappaB mediates diverse and paradoxical biological effects. PMID- 16046794 TI - Zebrafish and cardiac toxicology. AB - Model systems are a mainstay in toxicological research. Zebrafish are rapidly becoming an important model organism for studying vertebrate development. The advantages of zebrafish: short reproductive cycle, production of numerous transparent, synchronously developing embryos, low cost, and standardization make zebrafish an attractive model for toxicologists as well. The use of these fish to study heart development has moved forward very rapidly, laying the groundwork for studying the effects of chemicals on cardiac development and function. Here we describe approaches that can be used to study cardiac toxicity in developing zebrafish, focusing on examples where zebrafish embryos have been especially useful in understanding the impact of specific toxicants on heart development and function. PMID- 16046795 TI - Inhibition of neovascularization by environmental agents. AB - The formation of new blood vessels, neovascularization, occurs by two unique processes: vasculogenesis, the de novo assembly of blood vessels from angioblast precursors, and angiogenesis, the formation of new capillary sprouts from preexisting vessels. There are many potential targets by which environmental pollutants may inhibit neovascularization and thus there are many possible phenotypic outcomes. Two examples of environmental pollutants that have been demonstrated to inhibit neovascularization include 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD), a prototypical halogenated aromatic hydrocarbon, and constituents found in environmental tobacco smoke. Studies have shown that TCDD disrupts neoangiogenesis by inhibiting the expression of angiogenic stimuli as well as by reducing the responsiveness of endothelial cells to those stimuli. Additionally, studies have shown that constituents of environmental tobacco smoke, including pyradine and pyrazine derivatives, can potently inhibit the angiogenic process of branching as well as the vasculogenic process involved in capillary plexus formation. Further, the inhibition of neovascularization by either TCDD or environmental tobacco smoke constituents is associated with reduced endothelial cell proliferation and altered expression of extracellular matrix proteins. Future research that identifies the specific angiogenic signaling pathways that are disrupted by these pollutants will improve our ability to assess their risk to human health. Finally, it is likely that many other environmental pollutants impact neovascularization; however, very few have been studied in sufficient detail. Thus, additional research also is needed to identify those environmental agents that mediate their toxicity by disrupting neovascularization. PMID- 16046796 TI - Characterization of spontaneous and chemically induced cardiac lesions in rodent model systems: the national toxicology program experience. AB - Induction of heart disease can be related to exposure to a number of agents, including environmental chemicals. Studies with laboratory rodents are commonly used to identify cardiotoxic agents and to investigate mechanisms of toxicity. This study was conducted to characterize spontaneous and chemically-induced rodent heart lesions. A retrospective light-microscopic evaluation was performed on the hearts of F344 rats and B6C3F1 mice from National Toxicology Program studies of six chemicals in which chemically-induced myocardial toxicity was present: oxymetholone, monochloroacetic acid, 3,3'-4,4'- tetrachoroazoxybenzene, diethanolamine, urethane, and methyl bromide. Two myocardial lesions were observed: cardiomyopathy (multifocal myofiber degeneration that could occur spontaneously or as a treatment effect) and degeneration (diffuse myofiber degeneration that was clearly related to treatment). Oxymetholone produced cardiotoxicity that was apparent as an increase in the incidence and average severity of cardiomyopathy. The remaining five chemicals produced degeneration, which appeared morphologically similar with each of the chemicals. Based on available information concerning possible mechanisms by which each of these chemicals may induce cardiotoxicity, this evaluation indicated it may be possible to place the chemicals into two main categories: (1) those that primarily affected the coronary vasculature with secondary effects on the myocardium (oxymetholone), and (2) those that had a direct toxic effect on the myocardial cells (the remaining five chemicals). Beyond this, however, light-microscopic findings did not indicate any specific mechanisms. Additional morphologic evaluations, such as electron microscopy or special histochemical or immunostains, may help identify specific subcellular sites of toxic damage, which in turn can indicate appropriate types of molecular mechanistic studies. PMID- 16046800 TI - Data requirements for protein identification using chemically-assisted fragmentation and tandem mass spectrometry. AB - Many laboratories identify proteins by searching tandem mass spectrometry data against genomic or protein sequence databases. These database searches typically use the measured peptide masses or the derived peptide sequence and, in this paper, we focus on the latter. We study the minimum peptide sequence data requirements for definitive protein identification from protein sequence databases. Accurate mass measurements are not needed for definitive protein identification, even when a limited amount of sequence data is available for searching. This information has implications for the mass spectrometry performance (and cost), data base search strategies and proteomics research. PMID- 16046801 TI - Chemically-assisted fragmentation combined with multi-dimensional liquid chromatography and matrix-assisted laser desorption/ionization post source decay, matrix-assisted laser desorption/ionization tandem time-of flight or matrix assisted laser desorption/ionization tandem mass spectrometry for improved sequencing of tryptic peptides. AB - Derivatization of tryptic peptides using an Ettan CAF matrix-assisted laser desorption/ionization (MALDI) sequencing kit in combination with MALDI-post source decay (PSD) is a fast, accurate and convenient way to obtain de novo or confirmative peptide sequencing data. CAF (chemically assisted fragmentation) is based on solid-phase derivatization using a new class of water stable sulfonation agents, which strongly improves PSD analysis and simplifies the interpretation of acquired spectra. The derivatization is performed on solid supports, ZipTip(microC18, limiting the maximum peptide amount to 5 microg. By performing the derivatization in solution enabled the labeling of tryptic peptides derived from 100 microg of protein. To increase the number of peptides that could be sequenced, derivatized peptides were purified using multidimensional liquid chromatography (MDLC) prior to MALDI sequencing. Following the first dimension strong cation exchange (SCX) chromatography step, modified peptides were separated using reversed-phase chromatography (RPC). During the SCX clean up step, positively charged peptides are retained on the column while properly CAF derivatized peptides (uncharged) are not. A moderately complex tryptic digest, prepared from six different proteins of equimolar amounts, was CAF-derivatized and purified by MDLC. Fractions from the second dimension nano RPC step were automatically sampled and on-line dispensed to MALDI sample plates and analyzed using MALDI mass spectrometry fragmentation techniques. All proteins in the derivatized protein mixture digest were readily identified using MALDI-PSD or MALDI tandem mass spectrometry (MS/MS). More than 40 peptides were unambiguously sequenced, representing a seven-fold increase in the number of sequenced peptides in comparison to when the CAF-derivatized protein mix digest was analyzed directly (no MDLC-separation) using MALDI-PSD. In conclusion, MDLC purification of CAF-derivatized peptides significantly increases the success rate for de novo and confirmative sequencing using various MALDI fragmentation techniques. This new approach is not only applicable to single protein digests but also to more complex digests and could, thus, be an alternative to electrospray ionization MS/MS for peptide sequencing. PMID- 16046802 TI - A tandem time-of-flight mass spectrometer: combination of a multi-turn time-of flight and a quadratic-field mirror. AB - A tandem time-of-flight (ToF) mass spectrometer consisting of a multi-turn time of-flight (ToF) and a quadratic-field ion mirror has been designed and constructed. The instrument combines the unique capabilities of both ToF instruments, namely high-resolution and monoisotopic precursor ion selection from the multi-turn ToF and temporal focus for fragment ions with different kinetic energies from the quadratic-field mirror. The first tandem mass spectra for this unique combination of ToF systems are presented. PMID- 16046803 TI - Advantages and drawbacks of popular supercritical fluid chromatography/mass interfacing approaches--a user's perspective. AB - Mobile phases in supercritical fluid chromatography (SFC) have low viscosities and high diffusion coefficients with respect to those of traditional high performance liquid chromatography (HPLC). These properties allow higher mobile phase flow rates and/or longer columns in SFC, resulting in rapid analyses and high efficiency separations. In addition, chiral SFC is becoming especially popular. Mass spectrometry (MS) is arguably the most popular "informative" detector for chromatographic separations. Most SFC/MS is performed with atmospheric pressure ionization (API) sources. Unlike LC/MS, the interface between the SFC column and the API source must allow control of the downstream (post-column) pressure while also providing good chromatographic fidelity. Here we compare and contrast the popular interfacing approaches. Some are simple, such as direct effluent introduction with no active back-pressure-regulator (BPR) in high speed bioanalytical applications. The pressure-regulating-fluid interface is more versatile and provides excellent chromatographic fidelity, but is less user friendly. The pre-BPR- split interface and an interface which provides total-flow introduction with a mechanical BPR are good compromises between user friendliness and performance, and have become the most popular among practitioners. Applications of SFC/MS using these various interfaces are also discussed. PMID- 16046804 TI - On-line solid phase extraction using the Prospekt-2 coupled with a liquid chromatography/tandem mass spectrometer for the determination of dextromethorphan, dextrorphan and guaifenesin in human plasma. AB - An on-line liquid chromatography/tandem mass spectrometry (LC-MS/MS) procedure, using the Prospekt- 2 system, was developed and used for the determination of the levels of the active ingredients of cough/cold medications in human plasma matrix. The experimental configuration allows direct plasma injection by performing on- line solid phase extraction (SPE) on small cartridge columns prior to elution of the analyte(s) onto the analytical column and subsequent MS/MS detection. The quantitative analysis of three analytes with differing polarities, dextromethorphan (DEX), dextrorphan (DET) and guaifenesin (GG) in human plasma presented a significant challenge. Using stable-isotope-labeled internal standards for each analyte, the Prospekt-2 on-line methodology was evaluated for sensitivity, suppression, accuracy, precision, linearity, analyst time, analysis time, cost, carryover and ease of use. The lower limit of quantitation for the on line SPE procedure for DEX, DET and GG was 0.05, 0.05 and 5.0 ng mL(-1), respectively, using a 0.1 mL sample volume. The linear range for DEX and DET was 0.05-50 ng mL(-1) and was 5-5,000 ng mL(-1) for GG. Accuracy and precision data for five different levels of QC samples were collected over three separate days. Accuracy ranged from 90% to 112% for all three analytes, while the precision, as measured by the %RSD, ranged from 1.5% to 16.0% PMID- 16046805 TI - Development of an ion-pair reverse-phase liquid chromatographic/tandem mass spectrometry method for the determination of an 18-mer phosphorothioate oligonucleotide in mouse liver tissue. AB - A quantitative method for the determination of a partially modified, 2'-ribose alkoxy 18-mer phosphorothioate oligonucleotide, in liver tissue has been developed. A liquid:liquid extraction, ion-pair reverse phase chromatographic separation, and tandem mass spectrometry were used to achieve a quantitation range of 125 to 10,000 ng g(-1) mouse liver tissue. A total cycle time of 5 min was obtained while maintaining separation of three potential impurities. Separations were performed using a Discovery RP-Amide C16, 100 x 2 mm column packed with 5 microm particles. The separation was facilitated by the use of triethylamine (TEA) and hexafluoroisopropanol (HFIP) as ion-pair agents. The method has subsequently been used for the determination of other phosphorothioate oligonucleotides in support of discovery research. PMID- 16046806 TI - Elucidation of tautomer structures of 2-acetyltetrahydropyridine using gas chromatography/mass spectrometry and gas chromatography/infrared spectroscopy. AB - 2-Acetyltetrahydropyridine is an important flavor component in heated corn products such as popcorn and corn chips. The compound exists as a mixture of two tautomers that have different flavor/aroma characteristics. The tautomers also exhibit different chromatographic behaviors and are distinguishable spectroscopically. Though their electron ionization mass spectra are different, structure assignment based on low-resolution mass spectra alone has been subject to error. A combination of high resolution exact mass measurement and vapor phase infrared measurement of the separated tautomers was used to unambiguously assign the tautomer structures. PMID- 16046807 TI - Shotgun sequencing small oligonucleotides by nozzle-skimmer dissociation and electrospray ionization mass spectrometry. AB - Nozzle-skimmer dissociation in combination with de novo sequencing was investigated as an approach for increasing the throughput of oligonucleotide analysis attainable by electrospray ionization mass spectrometry. An experimental method allowing for the sequential generation of precursor and fragment ion data during direct infusion of sample was developed. These data can then be used with readily available de novo sequencing software to characterize small oligonucleotides. When this approach was applied to mixtures of oligonucleotides, it was found that de novo sequencing becomes limited due to spectral congestion and overlapping oligonucleotide m/z dissociation product values. Self-packed C(18) microspray emitters were investigated as a means of reducing spectral complexity. It was found that such emitters allow for the analysis of oligonucleotide mixtures with minimal component overlap, and these emitters provide additional benefits of pre- concentrating and desalting the sample. These developments can provide a route for the more rapid characterization of ribonucleic acid endonuclease digestion mixtures. PMID- 16046808 TI - Chiral and isomeric analysis by electrospray ionization and sonic spray ionization using the fixed-ligand kinetic method. AB - The fixed-ligand version of the kinetic method has been used for chiral and for isomeric analysis by studying the dissociation kinetics of transition metal-bound trimeric cluster ions ([(M(II) + L(fixed)-H)(ref*)(An)](+), where M(II) is a transition metal, L fixed is a fixed (non-dissociating) ligand, ref* is a reference ligand and An is the analyte. The trimeric cluster ions are readily generated by electrospray ionization (ESI) or sonic spray ionization (SSI). The size of the fixed ligand, L- Phe-Gly-L-P he-Gly, is chosen based on previous results but with the inclusion of aromatic functionality to increase chiral recognition. Improved chiral/isomeric differentiation results from enhanced chiral/isomeric interactions (metal-ligand and ligand-ligand) due to the fixed ligand. As shown in the cases of chiral dipeptides (D-Ala-D-Ala/L-Ala-L-Ala), sugars (D/L-glucose, D/L-mannose) and isomeric tetrapeptides (L-Ala-Gly-Gly Gly/Gly-Gly -Gly-L-Ala), improved chiral/isomeric discrimination by factors from three to six were obtained by the fixed ligand procedure. Chiral recognition is independent of the concentrations of the analyte, the reference ligand, the fixed ligand and the transition metal salt, a great advantage for practical applications. In addition to increased chiral distinction, the simplified dissociation kinetics also contribute to improved accuracy in chiral quantification, in comparison with data obtained by investigating the dissociation kinetics of simple trimeric cluster ions [M(II)(ref*)2(An) H](+). Accurate determination of enantiomeric excess (ee) is demonstrated by enantiomeric quantification of D-Ala-D-Ala/L-Ala-L-Ala down to 2% ee. Both ESI and SSI allow chiral quantification with similar accuracies. The performance of chiral analysis experiments is not limited to ion trapping devices such as quadrupole ion trap mass spectrometers by a hybrid quadrupole-time of flight (Q ToF) mass spectrometer is shown to provide an alternative choice. The fixed ligand kinetic method is not restricted to any particular kinds of isomers and, hence, represents a general procedure for improving molecular recognition and chiral analysis in the gas phase. PMID- 16046809 TI - Tandem mass spectrometry characteristics of polyester anions and cations formed by electrospray ionization. AB - Electrospray ionization of polyesters composed of isophthalic acid and neopentyl glycol produces carboxylate anions in negative mode and mainly sodium ion adducts in positive mode. A tandem mass spectrometry (MS/MS) study of these ions in a quadrupole ion trap shows that the collisionally activated dissociation pathways of the anions are simpler than those of the corresponding cations. Charge-remote fragmentations predominate in both cases, but the spectra obtained in negative mode are devoid of the complicating cation exchange observed in positive mode. MS/MS of the Na(+) adducts gives rise to a greater number of fragments but not necessarily more structural information. In either positive or negative mode, polyester oligomers with different end groups fragment by similar mechanisms. The observed fragments are consistent with rearrangements initiated by the end groups. Single-stage ESI mass spectra also are more complex in positive mode because of extensive H/Na substitutions; this is also true for matrix-assisted laser desorption ionization (MALDI) mass spectra. Hence, formation and analysis of anions might be the method of choice for determining block length, end group structure and copolymer sequence, provided the polyester contains at least one carboxylic acid end group that is ionizable to anions. PMID- 16046810 TI - Data mining in genomics and proteomics. PMID- 16046811 TI - Early-stage folding in proteins (in silico) sequence-to-structure relation. AB - A sequence-to-structure library has been created based on the complete PDB database. The tetrapeptide was selected as a unit representing a well-defined structural motif. Seven structural forms were introduced for structure classification. The early-stage folding conformations were used as the objects for structure analysis and classification. The degree of determinability was estimated for the sequence-to-structure and structure-to-sequence relations. Probability calculus and informational entropy were applied for quantitative estimation of the mutual relation between them. The structural motifs representing different forms of loops and bends were found to favor particular sequences in structure-to-sequence analysis. PMID- 16046812 TI - Functional clustering algorithm for high-dimensional proteomics data. AB - Clustering proteomics data is a challenging problem for any traditional clustering algorithm. Usually, the number of samples is largely smaller than the number of protein peaks. The use of a clustering algorithm which does not take into consideration the number of features of variables (here the number of peaks) is needed. An innovative hierarchical clustering algorithm may be a good approach. We propose here a new dissimilarity measure for the hierarchical clustering combined with a functional data analysis. We present a specific application of functional data analysis (FDA) to a high-throughput proteomics study. The high performance of the proposed algorithm is compared to two popular dissimilarity measures in the clustering of normal and human T-cell leukemia virus type 1 (HTLV-1)-infected patients samples. PMID- 16046813 TI - Objective clustering of proteins based on subcellular location patterns. AB - The goal of proteomics is the complete characterization of all proteins. Efforts to characterize subcellular location have been limited to assigning proteins to general categories of organelles. We have previously designed numerical features to describe location patterns in microscope images and developed automated classifiers that distinguish major subcellular patterns with high accuracy (including patterns not distinguishable by visual examination). The results suggest the feasibility of automatically determining which proteins share a single location pattern in a given cell type. We describe an automated method that selects the best feature set to describe images for a given collection of proteins and constructs an effective partitioning of the proteins by location. An example for a limited protein set is presented. As additional data become available, this approach can produce for the first time an objective systematics for protein location and provide an important starting point for discovering sequence motifs that determine localization. PMID- 16046814 TI - High-betweenness proteins in the yeast protein interaction network. AB - Structural features found in biomolecular networks that are absent in random networks produced by simple algorithms can provide insight into the function and evolution of cell regulatory networks. Here we analyze "betweenness" of network nodes, a graph theoretical centrality measure, in the yeast protein interaction network. Proteins that have high betweenness, but low connectivity (degree), were found to be abundant in the yeast proteome. This finding is not explained by algorithms proposed to explain the scale-free property of protein interaction networks, where low-connectivity proteins also have low betweenness. These data suggest the existence of some modular organization of the network, and that the high-betweenness, low-connectivity proteins may act as important links between these modules. We found that proteins with high betweenness are more likely to be essential and that evolutionary age of proteins is positively correlated with betweenness. By comparing different models of genome evolution that generate scale-free networks, we show that rewiring of interactions via mutation is an important factor in the production of such proteins. The evolutionary and functional significance of these observations are discussed. PMID- 16046815 TI - Data-mining analysis suggests an epigenetic pathogenesis for type 2 diabetes. AB - The etiological origin of type 2 diabetes mellitus (T2DM) has long been controversial. The body of literature related to T2DM is vast and varied in focus, making a broad epidemiological perspective difficult, if not impossible. A data-mining approach was used to analyze all electronically available scientific literature, over 12 million Medline records, for "objects" such as genes, diseases, phenotypes, and chemical compounds linked to other objects within the T2DM literature but were not themselves within the T2DM literature. The goal of this analysis was to conduct a comprehensive survey to identify novel factors implicated in the pathology of T2DM by statistically evaluating mutually shared associations. Surprisingly, epigenetic factors were among the highest statistical scores in this analysis, strongly implicating epigenetic changes within the body as causal factors in the pathogenesis of T2DM. Further analysis implicates adipocytes as the potential tissue of origin, and cytokines or cytokine-like genes as the dysregulated factor(s) responsible for the T2DM phenotype. The analysis provides a wealth of literature supporting this hypothesis, which-if true-represents an important paradigm shift for researchers studying the pathogenesis of T2DM. PMID- 16046816 TI - Combining information from multiple data sources to create multivariable risk models: illustration and preliminary assessment of a new method. AB - A common practice of metanalysis is combining the results of numerous studies on the effects of a risk factor on a disease outcome. If several of these composite relative risks are estimated from the medical literature for a specific disease, they cannot be combined in a multivariate risk model, as is often done in individual studies, because methods are not available to overcome the issues of risk factor colinearity and heterogeneity of the different cohorts. We propose a solution to these problems for general linear regression of continuous outcomes using a simple example of combining two independent variables from two sources in estimating a joint outcome. We demonstrate that when explicitly modifying the underlying data characteristics (correlation coefficients, standard deviations, and univariate betas) over a wide range, the predicted outcomes remain reasonable estimates of empirically derived outcomes (gold standard). This method shows the most promise in situations where the primary interest is in generating predicted values as when identifying a high-risk group of individuals. The resulting partial regression coefficients are less robust than the predicted values. PMID- 16046817 TI - Contextual multiple sequence alignment. AB - In a recently proposed contextual alignment model, efficient algorithms exist for global and local pairwise alignment of protein sequences. Preliminary results obtained for biological data are very promising. Our main motivation was to adopt the idea of context dependency to the multiple-alignment setting. To this aim the relaxation of the model was developed (we call this new model averaged contextual alignment) and a new family of amino acids substitution matrices are constructed. In this paper we present a contextual multiple-alignment algorithm and report the outcomes of experiments performed for the BAliBASE test set. The contextual approach turned out to give much better results for the set of sequences containing orphan genes. PMID- 16046818 TI - Selecting genes by test statistics. AB - Gene selection is an important issue in analyzing multiclass microarray data. Among many proposed selection methods, the traditional ANOVA F test statistic has been employed to identify informative genes for both class prediction (classification) and discovery problems. However, the F test statistic assumes an equal variance. This assumption may not be realistic for gene expression data. This paper explores other alternative test statistics which can handle heterogeneity of the variances. We study five such test statistics, which include Brown-Forsythe test statistic and Welch test statistic. Their performance is evaluated and compared with that of F statistic over different classification methods applied to publicly available microarray datasets. PMID- 16046819 TI - Protein coding sequence identification by simultaneously characterizing the periodic and random features of DNA sequences. AB - Most codon indices used today are based on highly biased nonrandom usage of codons in coding regions. The background of a coding or noncoding DNA sequence, however, is fairly random, and can be characterized as a random fractal. When a gene-finding algorithm incorporates multiple sources of information about coding regions, it becomes more successful. It is thus highly desirable to develop new and efficient codon indices by simultaneously characterizing the fractal and periodic features of a DNA sequence. In this paper, we describe a novel way of achieving this goal. The efficiency of the new codon index is evaluated by studying all of the 16 yeast chromosomes. In particular, we show that the method automatically and correctly identifies which of the three reading frames is the one that contains a gene. PMID- 16046820 TI - Classification and selection of biomarkers in genomic data using LASSO. AB - High-throughput gene expression technologies such as microarrays have been utilized in a variety of scientific applications. Most of the work has been done on assessing univariate associations between gene expression profiles with clinical outcome (variable selection) or on developing classification procedures with gene expression data (supervised learning). We consider a hybrid variable selection/classification approach that is based on linear combinations of the gene expression profiles that maximize an accuracy measure summarized using the receiver operating characteristic curve. Under a specific probability model, this leads to the consideration of linear discriminant functions. We incorporate an automated variable selection approach using LASSO. An equivalence between LASSO estimation with support vector machines allows for model fitting using standard software. We apply the proposed method to simulated data as well as data from a recently published prostate cancer study. PMID- 16046821 TI - Gene expression data classification with Kernel principal component analysis. AB - One important feature of the gene expression data is that the number of genes M far exceeds the number of samples N. Standard statistical methods do not work well when N < M. Development of new methodologies or modification of existing methodologies is needed for the analysis of the microarray data. In this paper, we propose a novel analysis procedure for classifying the gene expression data. This procedure involves dimension reduction using kernel principal component analysis (KPCA) and classification with logistic regression (discrimination). KPCA is a generalization and nonlinear version of principal component analysis. The proposed algorithm was applied to five different gene expression datasets involving human tumor samples. Comparison with other popular classification methods such as support vector machines and neural networks shows that our algorithm is very promising in classifying gene expression data. PMID- 16046822 TI - Multiclass cancer classification by using fuzzy support vector machine and binary decision tree with gene selection. AB - We investigate the problems of multiclass cancer classification with gene selection from gene expression data. Two different constructed multiclass classifiers with gene selection are proposed, which are fuzzy support vector machine (FSVM) with gene selection and binary classification tree based on SVM with gene selection. Using F test and recursive feature elimination based on SVM as gene selection methods, binary classification tree based on SVM with F test, binary classification tree based on SVM with recursive feature elimination based on SVM, and FSVM with recursive feature elimination based on SVM are tested in our experiments. To accelerate computation, preselecting the strongest genes is also used. The proposed techniques are applied to analyze breast cancer data, small round blue-cell tumors, and acute leukemia data. Compared to existing multiclass cancer classifiers and binary classification tree based on SVM with F test or binary classification tree based on SVM with recursive feature elimination based on SVM mentioned in this paper, FSVM based on recursive feature elimination based on SVM can find most important genes that affect certain types of cancer with high recognition accuracy. PMID- 16046823 TI - Computational, integrative, and comparative methods for the elucidation of genetic coexpression networks. AB - Gene expression microarray data can be used for the assembly of genetic coexpression network graphs. Using mRNA samples obtained from recombinant inbred Mus musculus strains, it is possible to integrate allelic variation with molecular and higher-order phenotypes. The depth of quantitative genetic analysis of microarray data can be vastly enhanced utilizing this mouse resource in combination with powerful computational algorithms, platforms, and data repositories. The resulting network graphs transect many levels of biological scale. This approach is illustrated with the extraction of cliques of putatively co-regulated genes and their annotation using gene ontology analysis and cis regulatory element discovery. The causal basis for co-regulation is detected through the use of quantitative trait locus mapping. PMID- 16046824 TI - Online analytical processing (OLAP): a fast and effective data mining tool for gene expression databases. AB - Gene expression databases contain a wealth of information, but current data mining tools are limited in their speed and effectiveness in extracting meaningful biological knowledge from them. Online analytical processing (OLAP) can be used as a supplement to cluster analysis for fast and effective data mining of gene expression databases. We used Analysis Services 2000, a product that ships with SQLServer2000, to construct an OLAP cube that was used to mine a time series experiment designed to identify genes associated with resistance of soybean to the soybean cyst nematode, a devastating pest of soybean. The data for these experiments is stored in the soybean genomics and microarray database (SGMD). A number of candidate resistance genes and pathways were found. Compared to traditional cluster analysis of gene expression data, OLAP was more effective and faster in finding biologically meaningful information. OLAP is available from a number of vendors and can work with any relational database management system through OLE DB. PMID- 16046825 TI - Cardiovascular damage in Alzheimer disease: autopsy findings from the Bryan ADRC. AB - Autopsy information on cardiovascular damage was investigated for pathologically confirmed Alzheimer disease (AD) patients (n=84) and non-AD control patients (n=60). The 51 relevant items were entered into a grade-of-membership model to describe vascular damage in AD. Five latent groups were identified "I: early onset AD," "II: controls, cancer," "III: controls, extensive atherosclerosis," "IV: late-onset AD, male," and "V: late-onset AD, female." Expectedly, Groups IV and V had elevated APOE epsilon 4 frequency. Unexpectedly, there was limited atherosclerosis and frequent myocardial valve and ventricular damage. The findings do not indicate a strong relationship between atherosclerosis and AD, although both are associated with the APOE epsilon 4. Instead, autopsy findings of extensive atherosclerosis were associated with possible, not probable or definite AD, and premature death. They are consistent with the hypothesis that brain hypoperfusion contributes to dementia, possibly to AD pathogenesis, and raise the possibility that the APOE allele epsilon 4 contributes directly to heart valve and myocardial damage. PMID- 16046826 TI - Metabolite fingerprinting in transgenic Nicotiana tabacum altered by the Escherichia coli glutamate dehydrogenase gene. AB - With about 200,000 phytochemicals in existence, identifying those of biomedical significance is a mammoth task. In the postgenomic era, relating metabolite fingerprints, abundances, and profiles to genotype is also a large task. Ion analysis using Fourier transformed ion cyclotron resonance mass spectrometry (FT ICR-MS) may provide a high-throughput approach to measure genotype dependency of the inferred metabolome if reproducible techniques can be established. Ion profile inferred metabolite fingerprints are coproducts. We used FT-ICR-MS derived ion analysis to examine gdhA (glutamate dehydrogenase (GDH; EC 1.4.1.1)) transgenic Nicotiana tabacum (tobacco) carrying out altered glutamate, amino acid, and carbon metabolisms, that fundamentally alter plant productivity. Cause and effect between gdhA expression, glutamate metabolism, and plant phenotypes was analyzed by (13) NH(4)(+) labeling of amino acid fractions, and by FT-ICR-MS analysis of metabolites. The gdhA transgenic plants increased (13)N labeling of glutamate and glutamine significantly. FT-ICR-MS detected 2,012 ions reproducible in 2 to 4 ionization protocols. There were 283 ions in roots and 98 ions in leaves that appeared to significantly change abundance due to the measured GDH activity. About 58% percent of ions could not be used to infer a corresponding metabolite. From the 42% of ions that inferred known metabolites we found that certain amino acids, organic acids, and sugars increased and some fatty acids decreased. The transgene caused increased ammonium assimilation and detectable ion variation. Thirty-two compounds with biomedical significance were altered in abundance by GDH including 9 known carcinogens and 14 potential drugs. Therefore, the GDH transgene may lead to new uses for crops like tobacco. PMID- 16046827 TI - Finding groups in gene expression data. AB - The vast potential of the genomic insight offered by microarray technologies has led to their widespread use since they were introduced a decade ago. Application areas include gene function discovery, disease diagnosis, and inferring regulatory networks. Microarray experiments enable large-scale, high-throughput investigations of gene activity and have thus provided the data analyst with a distinctive, high-dimensional field of study. Many questions in this field relate to finding subgroups of data profiles which are very similar. A popular type of exploratory tool for finding subgroups is cluster analysis, and many different flavors of algorithms have been used and indeed tailored for microarray data. Cluster analysis, however, implies a partitioning of the entire data set, and this does not always match the objective. Sometimes pattern discovery or bump hunting tools are more appropriate. This paper reviews these various tools for finding interesting subgroups. PMID- 16046828 TI - Various morphologies of mitochondria. PMID- 16046829 TI - Role of the small leucine-rich proteoglycan (SLRP) family in pathological lesions and cancer cell growth. AB - The roles of lumican, a member of the small-leucine-rich-proteoglycan (SLRP) family, in pathological fibrosis, cancer tissues and tumor cell growth were reviewed. Lumican is predominantly localized in the areas of pathological fibrosis including the thickened intima of human coronary arteries, ischemic and reperfused hearts, and acute pancreatitis and chronic pancreatitis (CP)-like lesions adjacent to pancreatic cancer nests. In these lesions, lumican mRNA and protein were transiently and ectopically overexpressed in most of the vascular smooth muscle cells (VSMCs) that migrated into the thickened intima, myocardial cells adjacent to an ischemic lesion, acinar cells, islet cells and fibroblasts of pathological pancreatic tissues. The low expression level of lumican in breast cancer is associated with rapid progression and poor survival. Lumican mRNA in breast cancer is overexpressed in fibroblasts adjacent to cancer cells but not in cancer cells. Furthermore, the high expression level of lumican is associated with a high pathological tumor grade, a low estrogen receptor level in the cancer tissues, and young age of patients. The suppression of lumican expression in culture cells induces their cell growth. Lumican-transfected tumor cells are characterized by a strong suppression of their anchorage-independent growth and capacity of invasion. Lumican significantly suppressed subcutaneous tumor formation in syngenic mice, with a concomitant decrease in cyclin D1 expression level, and induced and/or enhanced the apoptosis of these cells. The autocrine mechanism in cancer cells and the paracrine mechanism in cancer cells and fibroblasts via transforming growth factor (TGF)-beta and Smad signals may play important roles in the regulation of tumor growth by SLRPs. PMID- 16046830 TI - Is complete resection of hypertrophic adenoma of the prostate possible with TURP? AB - Whether complete resection was possible with TURP was explained. A lot of adenoma remains after transurethral resection of the prostate (TURP), the other hand transurethral enucleation of the prostate (TUE) is useful for complete resection of an adenoma. PMID- 16046831 TI - Are sterile water and brushes necessary for hand washing before surgery in Japan? AB - PURPOSE: To examine whether sterile water and brushes are necessary for hand washing before surgery. METHOD: Twenty-two operating room nurses were randomly divided into two groups as follows: 11 nurses who used 7.5% povidone iodine (PVI group) and another 11 nurses who used 4% chlorhexidine gluconate (CHG group) to wash their hands using the rubbing method. All the nurses were examined for bacterial contamination of their hands before and after surgical hand rubbing. We used tap water to wash the hands at the sink used for washing surgical instruments in the operating room and non sterilized plastic brushes. RESULTS: No bacteria were detected in the tap water. Before washing the hands, the number of bacteria detected was 5.0 x 10(3) cfu/H in the PVI group and 4.0 x 10(3) cfu/H in the CHG group, which were similar in both groups. After washing the hands, the median value of the bacteria decreased to 8.7 x 10(2) cfu/H in the PVI group and 0 cfu/H in the CHG group. CONCLUSIONS: Sterile water and brushes are not necessary for preoperative scrubbing up. When using tap water for surgical hand washing, 1) the hand-rubbing method should be used; 2) a quick-alcohol-based disinfectant scrub should be used; 3) the concentration of free chloride in the water should be maintained at over 0.1 PPM; 4) the bacterial contamination of the water should be checked; and 5) the faucet should be routinely cleaned and sterilized. PMID- 16046832 TI - Re-expression of reduced VEGF activity in liver metastases of experimental pancreatic cancer. AB - PURPOSE: Vascular endothelial growth factor (VEGF) is thought to play a crucial role in the process of cancer growth and metastasis. In this study, the expression of VEGF in liver metastases of pancreatic cancer was investigated using an established hamster model. METHODS: Pancreatic cancer cells (PGHAM-1, 1 x 10(6)) derived from N-nitrosobis (2-oxopropyl) amine (BOP)-induced pancreatic tumors in Syrian golden hamsters were transplanted into the pancreas of female hamsters. All hamsters were sacrificed at 21 days after transplantation and used for the histopathological examination of pancreatic and metastatic lesions (primary transplantation model). The metastatic liver tumors were minced with scissors and 1 mm(3) tumors were retransplanted into the pancreas of a second hamster. All hamsters were sacrificed 21 days after retransplantation, and the pancreatic tumors were removed (back transplantation model). Immunohistochemical analyses using antibody against VEGF were performed for all pancreatic and liver tumors. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to examine the expression of VEGF mRNA in the tumors. In addition, we investigated the proliferation of each tumor using Ag-NOR staining. RESULTS: In the primary transplantation models, VEGF expression in the pancreatic tumors was positive, but that in the liver metastases was only weakly positive or negative. On the other hand, VEGF expression in the pancreatic tumors that had developed from the retransplantation of the liver tumors (back transplantation model) was strongly positive. VEGF mRNA was expressed in the pancreatic tumors of both primary and back transplantation models. In the metastatic liver tumors of the primary transplantation model, VEGF mRNA was expressed in all cases, although the immunohistochemical staining pattern was weakly positive or negative. Similarly, in the metastatic liver tumor of the back transplantation model, VEGF mRNA was expressed in all cases, although the immunohistochemical staining pattern was weakly positive or negative. No significant differences in Ag-NOR scores were found between the models. CONCLUSION: Our results suggest that VEGF expression usually occurs in PGHAM-1 cells but that VEGF expression is reduced during the process of liver metastasis and revived by retransplantation. Thus, the interrelationship between cancer cells and the organ environment might play an important role in VEGF expression. PMID- 16046833 TI - Electromyographic functional analysis of the lumbar spinal muscles with low back pain. AB - The lumbar paraspinal muscles were examined by surface electromyography (EMG) in 22 patients with low back pain and 22 healthy volunteers. Surface electrodes were placed bilaterally on the lumbar multifidus and longissimus muscles at the level of the spinous process of the third lumbar vertebra. Kinesiologic EMG was recorded, in standing resting position, during the following trunk motions: trunk forward flexion and extension, lateral bending and axial rotation. No muscular activity was observed in the full trunk flexion position in the control group. In contrast, continuous muscle activity was observed in the low back pain group. On axial rotation, an intermuscular time lag was observed at the beginning of the motion in the control group. In the low back pain group, there was no such time lag. Paraspinal muscle activities restricted lumbar range of motion, and protect from injury for movement. This suggests their role as stabilizers. PMID- 16046834 TI - Finite element analysis of undermining of pressure ulcer with a simple cylinder model. AB - With pressure sores, surface damage indicates that subcutaneous fat tissue necrosis has occurred. We hypothesized that formation of necrosis under a pressure sore changes the stress distribution, which in turn affects further extension of the necrosis. In the present study, two-dimensional finite element models were used to perform analysis under different undermining size conditions. Greater stress concentration was observed in the larger undermining model. This may be the reason that, in clinical situations, a large area of undermining necrosis is sometimes observed under the skin of sores with a small area of damage. PMID- 16046835 TI - Use of a bioabasorbable bone pin fixation for nasal fractures. AB - After repairing a nasal fracture, it is customary to provide postoperative support for the nasal bone with intranasal, antibiotic-soaked gauze packing and an external splint. However, in cases of a skull base fracture with liquorrhea, this procedure is generally contraindicated because of the risk of infection. To lessen this risk, the authors used a bioabsorbable pin which is ultimately absorbed by the body through the natural process of hydrolysis. An intranasal support was inserted from outside the skin down to the maxilla. The operative results were satisfactory with no postoperative complications such as an inflammatory reaction or an infection. PMID- 16046836 TI - Abdominal incisional hernia repair using the Composix Kugel Patch: two case reports. AB - We describe two patients with abdominal incisional hernias, which occurred after appendectomy and replacement of an artificial blood vessel. Both were treated by operative hernial repair with the Composix Kugel Patch (C.R. Bard Inc.), a composite mesh that combines polypropylene mesh and expanded polytetrafluoroethylene (Gore-Tex). The mesh has various beneficial characteristics. It is a reinforcing material for the abdominal wall; even when in direct contact with the intestinal tract it is minimally adherent to the intestinal tract. The mesh expands readily and is easily fixed to the abdominal wall because it has a shape-memory ring. The long-term results of operative repair with this mesh have not yet been reported, but it is hoped that the aforementioned characteristics will yield favorable outcomes. PMID- 16046837 TI - Case report of metachronous hepatocellular carcinoma and early esophageal cancer. AB - We report a case of double cancer of the liver and esophagus, an extremely rare condition that can be very difficult to treat. A tumorous lesion was pointed out in segment 6 of the liver of a 69-year-old man under treatment for liver cirrhosis. Abdominal computed tomography and angiography revealed a hepatocellular carcinoma. The patient underwent transcatheter arterial embolization and partial resection of segments 5 and 6. Fourteen months later, a small elevated lesion was detected in the esophagus during an endoscopic examination. The patient was treated by endoscopic mucosal resection and radiation therapy at a total dose of 50 Gy. Histological examination revealed a squamous cell carcinoma with cancer cells confined within the epithelium of the esophagus. Over the 6 years since the hepatectomy, there have been no signs of recurrence. We report a successful curative resection in an extremely rare form of double cancer with a poor prognosis. PMID- 16046838 TI - Estrus synchronization and conception rate after a progesterone releasing intravaginal device (PRID) treatment from the early luteal phase in heifers. AB - The objective of the present study was to evaluate estrus synchronization and conception rate after progesterone releasing intravaginal device (PRID) treatment from the early luteal phase in the presence or absence of estradiol benzoate (EB) in heifers. Heifers (n=11) were assigned randomly to two treatments; insertion of a PRID containing 1.55 g progesterone with a capsule attached including 10 mg EB (P+EB; n=6) and the PRID withdrawn the EB capsule (P-EB; n=5). The PRID was inserted into the vagina on Day 2 of the estrous cycle (Day 0 was the day of ovulation) and was left for 12 days. The proportion of heifers exhibiting standing estrus within 3 days after PRID removal was 83.3% (5/6) for the P+EB group, and 80.0% (4/5) for the P-EB group, respectively. Conception rate by artificial insemination on synchronized estrus was 80.0% (4/5) in the P+EB group, and 100% (4/4) in the P-EB treatment group, respectively. These results suggest that a PRID treatment from 2 days after ovulation for 12 days in the presence or absence of EB has an effect on the synchronization of estrus and produces a beneficial conception rate in heifers. PMID- 16046839 TI - Whey acidic protein (WAP) depresses the proliferation of mouse (MMT) and human (MCF-7) mammary tumor cells. AB - We previously reported that the enforced expression of exogenous whey acidic protein (WAP) significantly inhibited the proliferation of mouse mammary epithelial cells (HC11 and EpH4/H6 cells). This paper presents the first evidence that WAP also depresses the proliferation of mammary tumor cells from mouse (MMT cells) and human (MCF-7 cells). We established WAP-clonal MMT and MCF-7 cell lines, and confirmed the secretion of WAP from the WAP-clonal cells into culture medium. The enforced expression of WAP significantly inhibited the proliferation of MMT and MCF-7 cells in in vitro culture. FACScan analyses revealed that G0/G1 phase cell-cycle progression was disordered and elongated in the WAP-clonal MMT and MCF-7 cells compared to that of the control cells. The expression of cyclin D1 was significantly decreased in the WAP-clonal MMT and MCF-7 cells, suggesting that progression from the G1 to the S phase was delayed in the WAP-clonal cells. The present results indicate that WAP plays a negative regulatory role in the cell-cycle progression of mammary tumor cells via a paracrine mechanism. PMID- 16046840 TI - Perinatal hypoxic-ischemic brain damage: evolution of an animal model. AB - Early research in the Vannucci laboratory prior to 1981 focused largely on brain energy metabolism in the developing rat. At that time, there was no experimental model to study the effects of perinatal hypoxia-ischemia in the rodent, despite the tremendous need to investigate the pathophysiology of perinatal asphyxial brain damage in infants. Accordingly, we developed such a model in the postnatal day 7 rat, using a modification of the Levine preparation in the adult rat. Rat pups underwent unilateral common carotid artery ligation followed by exposure to systemic hypoxia (8% oxygen) at a constant temperature of 37 degrees C. Brain damage, seen histologically, was generally confined to the cerebral hemisphere ipsilateral to the arterial occlusion, and consisted of selective neuronal death or infarction, depending on the duration of the systemic hypoxia. Tissue injury was observed in the cerebral cortex, hippocampus, striatum, and thalamus. Subcortical and periventricular white matter injury was also observed. This model was originally described in the Annals of Neurology in 1981, and during the more than 20 years since that publication numerous investigations utilizing the model have been conducted in our laboratories as well as laboratories around the world. Cerebral blood flow and metabolic correlates have been fully characterized. Physiologic and pharmacologic manipulations have been applied to the model in search of neuroprotective strategies. More recently, molecular biologic alterations during and following the hypoxic-ischemic stress have been ascertained and the model has been adapted to the immature mouse for specific use in genetically altered animals. As predicted in the original article, the model has proven useful for the study of the short- and long-term effects of hypoxic ischemic brain damage on motor activity, behavior, seizure incidence, and the process of maturation in the brain and other organ systems. PMID- 16046841 TI - Hypoxia preconditioning in the brain. AB - Exposure to moderate hypoxia alone does not cause neuronal death as long as blood pressure and cerebral blood flow are maintained in mammals. In neonatal and adult mammals including rats and mice, carotid occlusion in combination with hypoxia produces neuronal death and brain infarction. However, preexposure to 8% oxygen for 3 h protects the brain and likely other organs of neonatal and adult rats against combined hypoxia-ischemia 24 h later. In this paper, the possible mechanisms of this so-called hypoxia-induced tolerance to ischemia is discussed. One mechanism likely involves hypoxia-inducible factor-1alpha (HIF-1alpha). HIF 1alpha is a transcription factor that - during hypoxia - binds with a second protein (HIF-1beta) in the nucleus to promoter elements in hypoxia-responsive target genes. This causes upregulation of HIF target genes including VEGF, erythropoietin, iNOS, glucose transporter-1, glycolytic enzymes, and many other genes to protect the brain against ischemia 24 h later. In addition, non-HIF pathways including MTF-1, Egr-1 and others act directly or indirectly on other target genes to also promote hypoxia-induced preconditioning. Hypoxia preconditioning can be mimicked by iron chelators like desferrioxamine and transition metals like cobalt chloride that inhibit prolyl hydroxylases, increase HIF-1alpha levels in the brain, and produce protection of the brain against combined hypoxia-ischemia 24 h later. This hypoxia preconditioning has potential clinical usefulness in protecting high-risk newborns or to provide protection prior to surgery. PMID- 16046842 TI - Morphogenesis of the dentate gyrus: what we are learning from mouse mutants. AB - The dentate gyrus is one of two locations with continuing neurogenesis in adult mammals. While the function of adult neurogenesis is unknown, it is believed that it is involved in learning and memory. For adult neurogenesis to occur, the dentate gyrus must maintain the appropriate precursor cell niche in the subgranular zone, which is likely to be dependent on the developmental mechanisms at play in forming the dentate gyrus. In this review, we graft a molecular framework onto the known neuroanatomic developmental plan by considering the phenotypes of several mouse mutants that have well characterized dentate gyrus developmental abnormalities. This effort reveals that there are at least six distinct developmental steps that need to occur in the formation of the dentate gyrus, which can be associated with specific gene defects: (1) defining the dentate neuroepithelium; (2) forming the primary radial glial scaffolding; (3) radial migration of granule neurons to form the primordial granule cell layer; (4) establishing the precursor pool in the hilus; (5) radial transformation of the tertiary matrix, and (6) differentiation of dentate granule cells. From this analysis, it is clear that some molecular pathways control multiple steps in the development of the dentate gyrus. For example the Wnt pathway (steps 1, 2, 4) and the chemokine receptor CXCR4 (steps 3, 4) are involved in multiple developmental steps, while the neuronal differentiation gene NeuroD (step 6) and the integrin signaling pathway (step 5) are involved only in discrete stages of the dentate gyrus morphogenesis. PMID- 16046843 TI - Investigation of cerebral development and injury in the prematurely born primate by magnetic resonance imaging and histopathology. AB - We summarize the preliminary results of brain histopathology and magnetic resonance imaging applied to a premature baboon model developed for evaluation of the pathogenesis and treatment of bronchopulmonary dysplasia. Cerebral development was assessed in 10 gestational control animals at time points of 125, 140 and 160 days of gestation (dg). On the basis of histopathology, conventional MRI and diffusion MRI, 125 dg is equivalent to 26-28 weeks of human gestation, 140 dg is equivalent to 30-32 weeks, and 160 dg is equivalent to term. Preliminary data are also presented for 33 experimental cases delivered at 125 dg, nursed for 2 weeks in an intensive care facility, and sacrificed at 139-140 dg. The commonest neuropathology in this cohort is white matter damage, manifest by reactive astrogliosis or activated microglia, and enlarged ventricular size. Subarachnoid, germinal matrix and intraventricular hemorrhages are also common. These preliminary results support the similarity of this model to the human preterm infant for both cerebral development and the pattern of cerebral injury. The prematurely born baboon appears an important model for the study of preterm human birth. PMID- 16046844 TI - A new model for determining the influence of age and sex on functional recovery following hypoxic-ischemic brain damage. AB - Stroke is a disorder affecting the lives of all age groups, and particularly those at the opposite ends of the age spectrum. It is generally believed that the immature brain is more resistant to damage resulting from a hypoxic/ischemic injury, and that it is also more 'plastic' in terms of its ability to recover. Evidence from our laboratory, and a host of others, has indicated, however, that the developing brain may in fact be more sensitive to injury resulting from hypoxia-ischemia. The question remains, however, whether the immature brain has a greater capacity for recovery. In order to determine the relative capability for functional recovery between age groups, a stroke model of comparable injury is required. This paper describes a new rodent model of ischemic injury allowing for comparisons of behavioral recovery spanning the spectrum of ages between newborn and the elderly. Endothelin-1, a potent vasoconstrictor, was stereotactically injected into the brains of 10-, 63-, and 180-day-old Wistar rats, immediately adjacent to the middle cerebral artery. Regionally, the cortex, caudate, and thalamus were most significantly affected, with sparing of the hippocampus. Pathologic assessment indicated a similar degree of injury across age groups affecting the territorial distribution of the middle cerebral artery, with a predominance of damage in the anterior sections of the cortex and caudate (p < 0.05), compared to the posterior sections including the cortex and thalamus. There were no regional differences in the extent of damage between age groups. Interestingly, however, there were significant differences between males and females regarding the overall extent of brain damage (p < 0.05), with males showing greater damage than females. In addition, there were significant regional differences in the extent of damage between males and females, particularly regarding cortical damage (p < 0.05), both anteriorly and posteriorly, and the caudate anteriorly (p < 0.05). Our findings provide an important new model for comparison of brain damage among the entire spectrum of ages affected by stroke. Importantly, this will allow for further investigations regarding both functional recovery and gender difference comparisons. This may have important ramifications for the development of therapeutic interventions that are age and gender specific. PMID- 16046845 TI - Induction of reproducible focal ischemic lesions in neonatal mice by photothrombosis. AB - Focal stroke during the perinatal and neonatal period is a significant cause of cognitive and behavioral deficits. Currently, the number of models available to study neonatal brain injury is limited and many are technically difficult to induce in neonatal rodents. We demonstrate a reproducible method to induce a focal ischemic injury in the cerebral cortex of neonatal mice that utilizes the principle of photothrombosis. Postnatal day 7 pups were anesthetized and systemically administered rose bengal (50 mg/kg). Permanent focal ischemia was induced in the medial frontal cortex and somatosensory cortex by irradiating surface blood vessels with a laser (532 nm). By placing a mask having an aperture of defined shape and size on the skull surface, we were able to reliably and reproducibly induce infarcts in discretely defined cortical regions. Further, we demonstrate explicit control of infarct volume by modifying the duration of laser exposure. This tool will provide a means for researchers to safely, easily and noninvasively induce reproducible ischemic lesions in specified regions of the neonatal cortex. PMID- 16046846 TI - Strain variability, injury distribution, and seizure onset in a mouse model of stroke in the immature brain. AB - Neonatal stroke is an important cause of neurologic morbidity and cerebral palsy. Recently, we have determined that in postnatal day 12 CD1 mice unilateral carotid ligation alone results in seizures and brain injury. We have shown that, in this model, seizure scores correlate with brain injury scores. We have applied this model to another strain of mice to assess strain-related differences in vulnerability to seizures and brain injury after unilateral carotid ligation. Under isoflurane anesthesia, unilateral right-sided carotid ligation was performed in postnatal day 12 C3HeB/FeJ mice followed by a 4-hour period of observation in a 35 degrees C incubator. Seizure scores and brain jury scores were assigned and compared to scores in mice receiving sham surgery. Timing of seizure onset and regional distribution of brain injury were compared in the CD1 and C3HeB/FeJ mice. Unilateral carotid ligation in postnatal day 12 C3HeB/FeJ mice resulted in seizure behavior and brain injury in some animals, with similar time to seizure onset and regional injury distribution, but affected a significantly smaller percentage of C3HeB/FeJ pups than that observed in postnatal day 12 CD1 mice, indicating strain-related vulnerability in this model. PMID- 16046847 TI - Neuronal injuries induced by perinatal hypoxic-ischemic insults are potentiated by prenatal exposure to lipopolysaccharide: animal model for perinatally acquired encephalopathy. AB - We developed an original rat model for neonatal brain lesions whereby we explored the sequential effects of infectious and hypoxic-ischemic aggressions. We investigated the influence of combined exposure to prenatal infection with neonatal hypoxic-ischemic insult. Infectious effect was produced by administrating lipopolysaccharide (LPS) intraperitoneally to pregnant rats starting on embryonic day 17. Hypoxia-ischemia (H/I) was induced in the pups at postnatal day 1 (P1) by ligature of the right common carotid artery followed by exposure to hypoxia (8% O(2)) for 3.5 h. Animals were randomized into four groups: (1) control group: pups born to mothers subjected to intraperitoneal saline injection; (2) LPS group: pups exposed in utero to LPS; (3) H/I group: pups exposed to postnatal hypoxia after ligation of the right carotid artery, and (4) H/I plus LPS group: in utero exposure to LPS followed by postnatal hypoxia after ligation of the right carotid artery. Neuropathological findings in pups examined at P3 and P8 showed that groups 2, 3, and 4 presented a pattern of neuronal injury similar to those characterized as 'selective neuronal necrosis' within the context of human perinatal encephalopathy. Neuronal cellular injuries were particularly seen in the neocortex, mainly in parasagittal areas. The extent of neuronal cell injury in the brain of rats exposed to postnatal H/I was significantly increased by antenatal exposure to LPS. This animal model provides an experimental means to explore the respective roles of anoxic and infectious components in the pathogenesis of perinatal brain lesions and consequent cerebral palsy. PMID- 16046848 TI - Combined deficiency of IL-1beta18, but not IL-1alphabeta, reduces susceptibility to hypoxia-ischemia in the immature brain. AB - Interleukin (IL)-1 and IL-18 belong to the IL-1 family. IL-18 deficiency has been shown to confer moderate protection after hypoxia-ischemia (HI) in the immature brain, while the contribution of the two isoforms of IL-1 (IL-1alpha and IL 1beta) in neonatal HI brain injury has not been investigated previously. The aim of this study was to examine the contribution of the different members of the IL 1 family to neonatal HI damage. Unilateral HI was induced at postnatal day 9 in IL-1beta, IL-1beta18, and IL-1alphabeta knockout and wild-type mice and brain injury was evaluated 1 week later. IL-1beta18-deficient mice showed 17% reduction in brain injury, while no significant reduction in injury was detected between any of the other groups. These results indicate that IL-18, but not IL-1beta, or the combination of IL-1alpha and IL-1beta, is a contributor to HI injury in the immature brain. PMID- 16046849 TI - Gray matter oligodendrocyte progenitors and neurons die caspase-3 mediated deaths subsequent to mild perinatal hypoxic/ischemic insults. AB - With significant improvements in neonatal care, fewer infants sustain severe injury as a consequence of hypoxia/ischemia (H/I). However, the majority of experimental studies have inflicted moderate to severe injuries, or they have assessed damage to the caudal forebrain; therefore, to better understand how a mild H/I episode affects the structures and cells of the rostral forebrain, we assessed the relative vulnerabilities of cells in the neocortex, striatum, corpus callosum, choroid plexus and subventricular zone (SVZ). To inflict mild H/I injury, the right common carotid artery was ligated followed by 1 h of hypoxia (8% O(2)) at 37 degrees C. Regional vulnerabilities were assessed using TUNEL, active caspase-3 and hematoxylin and eosin staining at 24 and 48 h of recovery. Scattered columns of cell death were observed in the neocortex with deep-layer neurons more vulnerable than more superficial neurons. The majority of these dying neurons appeared to be dying apoptotic rather than necrotic deaths. In addition, approximately 1/3 of the apoptotic cells in the neocortex were O4+ oligodendrocyte progenitors. We also observed a decrease in NG2 staining within the affected regions of the forebrain. By contrast, active caspase-3+/S-100beta+ astrocytes were not observed. Neurons and O4+ oligodendrocyte progenitors also died apoptotic deaths within the striatum. The lining cells of the choroid plexus also sustained damage. Elevated numbers of apoptotic cells were observed in the most lateral region of the SVZ and some of these dying cells were O4+. The most novel finding of this study, that oligodendrocyte progenitors in the gray matter are damaged and eliminated as a consequence of perinatal H/I, provides new insights into the histopathology and neurological deficits observed in infants who sustain mild H/I brain injuries. PMID- 16046850 TI - Patterns of excitotoxin-induced brain lesions in the newborn rabbit: a neuropathological and MRI correlation. AB - There is pressing need to employ new advances in structural MR brain imaging to better diagnose brain damage in newborn infants. Timely application of such technology will enable improved therapeutic interventions. Diffusion-weighted sequences are a sensitive marker of very early neuronal injury, the spatial pattern of which provides critical information regarding the underlying pathophysiology. We have modified our murine model of excitotoxic neonatal brain injury to the rabbit, an animal whose brain is larger and where the neuroanatomic organization of the subcortical white matter more closely resembles that of the human. Utilizing this rabbit model, we undertook an MRI/histopathologic correlation. We found that as with the mouse, there is a spatiotemporal selectivity to the pattern of brain injury, and that the period from postnatal day (P) 7 to P9 in rabbits corresponds to the time of maximum vulnerability of the brain to excitotoxic white matter damage, which neuropathologically simulates periventricular leukomalacia (PVL). We additionally noted that diffusion-weighted imaging provided the most sensitive means of detecting such lesions and that this method was sensitive to structural maturational changes accompanying the normal cortical ontogeny. Taken together, our findings suggest that this rabbit model of perinatal excitotoxic brain injury will be a valuable addition to experimental approaches to further our understanding of perinatal brain damage, that diffusion weighted imaging will be an invaluable adjunct to the diagnosis of such injury, and that therapeutic strategies aimed at interrupting the evolution of PVL should include targeting the pathophysiologic cascade induced by excitotoxic neonatal brain injury. PMID- 16046851 TI - Overexpression of glutathione peroxidase protects immature murine neurons from oxidative stress. AB - Neuronal enzyme systems involved in free radical detoxification are developmentally regulated such that intracellular glutathione peroxidase (GPx-1) activity is low in the newborn mouse brain. We hypothesized that neurons expressing a higher level of GPx-1 will be more resistant to hydrogen peroxide (H(2)O(2)) exposure. We show a dose-dependent protection against H(2)O(2) in primary neuronal cultures from fetuses overexpressing human GPx-1 compared to wild types of the same genetic background. Exogenous antioxidants completely protected neurons, even at extremely high H(2)O(2 )concentrations and regardless of the genotype. Specific depletion of glutathione with buthionine sulfoximine increased cell death in transgenic cultures exposed to 200 microM H(2)O(2), reducing protection afforded by increased GPx-1 activity. Increased GPx-1 expression in immature cortical neurons confers protection from oxidative stress, but availability of reducing equivalents determines susceptibility to oxidative cell death. PMID- 16046852 TI - Non-protein-bound iron in brain interstitium of newborn pigs after hypoxia. AB - Oxidative damage is implied in perinatal hypoxic-ischemic brain injury, most importantly in white matter. Nonprotein-bound iron (NPBI) catalyzes the formation of toxic hydroxyl radicals. We measured the extracellular level of NPBI through microdialysis in the cortex, striatum, and periventricular white matter before, during and after severe hypoxia in newborn pigs. NPBI was analyzed by a new spectrophotometric method in which ferrous iron is chelated by bathophenanthroline. NPBI was present in all brain areas under baseline conditions and increased in white matter from 0.97 (0.69) to 2.75 (1.85) micromol/l (not corrected for recovery rate) during early reoxygenation. NPBI may contribute to oxidative injury after perinatal hypoxic insults. PMID- 16046853 TI - Glycolysis and perinatal hypoxic-ischemic brain damage. AB - To ascertain the regulation of glycolysis during perinatal hypoxia-ischemia, 7 day postnatal rats were subjected to unilateral common carotid artery ligation followed by hypoxia with 8% oxygen for up to 90 min. Brain concentrations of glucose, lactate, and key glycolytic intermediates were determined at specific intervals of hypoxia. During hypoxia-ischemia, anaerobic glycolysis increased to approximately 62% of its maximal capacity, which equates to a 135% stimulation of the glycolytic flux. The key regulatory enzymes, hexokinase, phosphofructokinase and pyruvate kinase, were all stimulated during hypoxia-ischemia, and there were no enzymatic rate limitations. The major rate-limiting step for glycolysis was the transport of glucose across the blood-brain barrier into the brain. PMID- 16046854 TI - Brief induced hypothermia improves outcome after asphyxial cardiopulmonary arrest in juvenile rats. AB - The American Heart Association has endorsed the use of mild hypothermia for adults after cardiopulmonary arrest. However, there are no contemporary trials testing hypothermia in children after cardiopulmonary arrest and extrapolation from adult studies is problematic given differences in brain development and primary etiology (asphyxia in children vs. ventricular arrhythmia in adults). Accordingly, we tested the effects of mild postresuscitative hypothermia on functional and histopathological outcome after asphyxial cardiac arrest in juvenile rats. Postnatal day 17 rats were subjected to 8 min of asphyxia-induced cardiac arrest followed by resuscitation. Rats were randomized to normothermic (37 degrees C), hypothermic (32 degrees C), or unregulated temperature groups (n = 7-8/group) to begin after return of spontaneous circulation for a duration of 1 h. Brain temperature in the unregulated group dropped to 34.0 +/- 0.4 degrees C at 1 h. The hypothermic group had improved motor function assessed using beam balance and inclined plane tests vs. the normothermic group. The depth of hypothermia was associated with increased CA1 hippocampal neuron survival at 5 weeks. Neurodegeneration in the CA1 hippocampus assessed using Fluoro-Jade B labeling at 5 weeks was not detected in the 32 degrees C group, whereas 2/7 and 4/7 rats in the 34 and 37 degrees C groups, respectively, showed neurodegeneration. Brief treatment with moderate induced hypothermia improved functional outcome and prevented long-term neurodegeneration in a model that mimics the clinical and histopathological scenario of pediatric cardiac arrest. Similar to adults, infants and children may benefit from induced hypothermia after cardiopulmonary arrest, warranting further study. PMID- 16046855 TI - Melatonin provides neuroprotection in the late-gestation fetal sheep brain in response to umbilical cord occlusion. AB - Oxygen free radicals, including the highly toxic hydroxyl radical (*OH), initiate lipid peroxidation and DNA/RNA fragmentation and damage cells. The pineal hormone melatonin is an antioxidant and powerful scavenger of *OH. We hypothesized that maternally administered melatonin could reduce *OH formation, lipid peroxidation, and DNA/RNA damage in the fetal brain in response to asphyxia. In 15 fetal sheep, extracellular *OH was measured by microdialysis in white and gray matter of the parasagittal cortex. In 10 fetuses, asphyxia was induced by umbilical cord occlusion for 10 min using an inflatable cuff - the ewes of these fetuses received either intravenous melatonin (1 mg bolus, then 1 mg/h for 2 h; n = 5) or vehicle (1% ethanol in saline; n = 5), and results were compared to fetuses with sham cord occlusion and vehicle-infused ewes (n = 5). Hypoxemia, acidemia, hypertension and bradycardia produced by cord occlusion was similar in the melatonin- and vehicle-treated groups. In the vehicle-treated group, cord occlusion resulted in a significant increase in *OH in gray matter at 8-9.5 h after occlusion (p < 0.05); in contrast, there was no *OH change in the melatonin treated group. After cord occlusion, lipid peroxidation (4-hydroxynonenal immunoreactivity) found throughout the brain of vehicle-infused ewes was significantly less in the melatonin-infused group. Melatonin had no significant effect on the distribution of DNA/RNA fragmentation, as shown by 8 hydroxydeoxyguanosine immunoreactivity. Thus, brief asphyxia results in significant and delayed entry of *OH into the extracellular space of cortical gray matter in the fetal sheep brain, and melatonin given to the mother at the time of the insult abrogates this increase. Melatonin, in reducing O2 free radical production, may be an effective neuroprotective treatment for the fetus. PMID- 16046856 TI - Differential effects of hypoxia-ischemia on phosphorylation of the N-methyl-D aspartate receptor in one- and three-week-old rats. AB - The effects of transient cerebral hypoxia-ischemia (HI) on phosphorylation of the NR1 subunit of the N-methyl-D-aspartate (NMDA) receptor were investigated in 7 (P7)- and 21 (P21)-day-old rats. Unilateral HI was induced by ligation of the right common carotid artery and exposure to 8% O(2)/92% N(2) for 120 (P7) or 90 (P21) min. Phosphorylation by protein kinase A (PKA; S897) and PKC (S896 and S890) was depressed in the ipsilateral hemisphere relative to both naive controls and the contralateral hemisphere immediately following HI at both ages. At P7, but not P21, reperfusion resulted in an initial recovery to control phosphorylation levels at all 3 sites followed by a secondary decline. At both ages, pS896 was less than control values after 24 h of recovery, whereas pS890 had returned to control levels by this time. pS897 recovered to control levels by 24 h in P21 animals but not in P7 animals. Differential effects of HI on phosphorylation of the NMDA receptor at P7 and P21 may contribute to age-related changes in sensitivity to HI. PMID- 16046857 TI - Expression of erythropoietin and its receptor in the brain of late-gestation fetal sheep, and responses to asphyxia caused by umbilical cord occlusion. AB - Asphyxia and hypoxia are common threats faced by the fetus in utero. In late gestation fetal sheep, asphyxia produced by umbilical cord occlusion (UCO) results in widespread lipid peroxidation and apoptosis. Adaptive mechanisms that might limit fetal brain damage include induction of the hemopoietic cytokine, erythropoietin (EPO). In unanesthetized fetal sheep, we investigated if 1 or 2 bouts of brief asphyxia (UCO for 10 min) induced EPO and EPO type I receptor (EPO R) expressions, with the second UCO repeated 48 h after the first. Fetal brains were recovered 48 h after either sham, 1 x or 2 x UCO at 129-133 (term approximately 147) days of gestation and prepared for immunocytochemistry. In age matched control brain, low levels of EPO and EPO-R proteins were present in oligodendrocytes (OLs), periventricular and cortical white matter (WM), with no EPO and very low EPO-R expression in neurons. After 1 x UCO, EPO and EPO-R expressions were increased in astrocytes (periventricular and cortical WM, striatum, corpus callosum), choroid plexus epithelial cells, scattered neurons in cortical layers IV-VI, hippocampal CA1 neurons, and in the molecular and granule layers of the cerebellum. After 2 x UCO, higher levels of EPO and EPO-R occurred in the periventricular and cortical WM, corpus callosum, hippocampal CA1, and in neurons of all cortical layers. Paradoxically, EPO and EPO-R were now lower in hippocampal CA1 neurons and cerebellar molecular and granule cell layers. Few OLs expressed EPO or EPO-R after 1 x or 2 x UCO. Thus, brief asphyxia induces EPO and EPO-R in fetal astrocytes, but only after repeated asphyxial insult in neurons. Whether this is a response to increased injury, or represents an adaptive response that limits further cell death and brain damage awaits further investigation. PMID- 16046858 TI - Subventricular zone proliferation after alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid receptor-mediated neonatal brain injury. AB - The mammalian forebrain subventricular zone (SVZ) contains stem cells capable of generating new neurons and glia. Recent studies indicate that acute brain injury is a potent stimulus for SVZ stem cell proliferation. To better understand mechanisms of the SVZ response to neonatal brain injury, we used a model that focuses on a unique mechanism of vulnerability of the immature CNS, alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor-mediated excitotoxicity. We previously demonstrated that intracerebroventricular injection of the glutamate analog AMPA in rats at postnatal day 7 (P7) caused bilateral periventricular gray and white matter injury. We hypothesized that excitotoxic injury would stimulate cellular proliferation in the SVZ; we used the AMPA intracerebroventricular injection model to test this hypothesis. P7 rat pups received either left or right intracerebroventricular injections of S-AMPA (2.5 nmol). Normal and PBS-injected littermates were included as controls. On P8 or P14, serial coronal sections through the SVZ were collected; an immunohistochemical assay was performed with an antibody to the cell proliferation marker Ki-67. Bilateral Ki-67+ cells/SVZ were quantitated stereologically using the optical disector method. The median number of Ki-67+ cells/SVZ was increased in the SVZ of AMPA-injected rats relative to normal controls on both P8 and P14. To evaluate neurogenesis, we assayed the expression of doublecortin, a microtubular protein expressed only by immature neurons. From P8 to P14, there was a marked increase in doublecortin immunoreactive cells in the AMPA-injected SVZ. Many Ki-67+ nuclei were immediately surrounded by doublecortin staining. This study indicates that there is a proliferative response in the immature SVZ after an excitotoxic stimulus. Our findings suggest that some of these newly generated cells differentiate as immature neurons. This model may provide information about the mechanisms that regulate SVZ responses to neonatal brain injury. PMID- 16046859 TI - Augmented constitutive CREB expression in the nucleus accumbens and striatum may contribute to the altered behavioral response to cocaine of adult mice exposed to cocaine in utero. AB - Neuroadaptations occurring in the mesolimbic dopamine pathway following recurrent exposure to drugs of abuse have been correlated with a behavioral phenomenon known as behavioral sensitization. We have developed an animal model of prenatal cocaine exposure and, using a postnatal sensitization protocol, have examined the subsequent sensitivity of offspring to cocaine. Pregnant Swiss Webster dams were injected twice daily from embryonic day 8 to 17, inclusive, with cocaine (COC40: administered cocaine HCl at a dose of 40 mg/kg/day, and COC20: administered cocaine HCl at a dose of 20 mg/kg/day), or saline (SAL). The SPF40 group (saline pair-fed), a nutritional control group, was 'pair-fed' with COC40 dams. Activity was recorded for 30 min during a 3-day saline habituation, a 14-day 'initiation' phase, when animals received cocaine (15 mg/kg) or saline every other day, and following a 21-day 'withdrawal' period when all mice were challenged with cocaine. COC40 offspring, as compared with SAL controls, did not habituate to a novel environment, demonstrated increased cocaine-induced stereotypy on Coc 1 (first cocaine injection), and blunted locomotor sensitization on challenge as measured by the percentage of each animal's baseline locomotion. Tissue samples of the nucleus accumbens (NAc) and striatum (Str) of all four prenatal treatment groups were examined to determine whether alterations in the transcription factor CREB or glutamate receptor subunit, GluR1, induced by prenatal cocaine treatment may have contributed to the altered behavioral responses. Immunoblot quantitation revealed significantly increased constitutive CREB expression in the NAc and Str of COC40 mice as compared with SAL controls. Such alterations in constitutive CREB levels may contribute to some of the behavioral differences reported in adult mice exposed to cocaine in utero. PMID- 16046861 TI - Meckel's cartilage differentiation is dependent on hedgehog signaling. AB - The hedgehog (Hh) signaling pathway has been shown to be essential for craniofacial development. Although mandibular arch derivatives are largely absent in Shh null mice, little is known about the role of Hh signaling during Meckel's cartilage development per se. Mandible development is dependent on the morphogenesis of Meckel's cartilage, which then serves as a template for subsequent skeletal differentiation. In this study, we examine the biological function of Hh signaling during Meckel's cartilage development in vivo and in vitro. E13.5 Shh null mice present a small mesenchymal condensation in the region of a presumptive Meckel's cartilage in the hypoplastic mandibular arch. By E15.5, the Shh mutant exhibits a mere remnant of the mandibular arch, without evidence of Meckel's cartilage differentiation. Further, wild-type embryonic (E11 or E12) mandibular explants cultured for up to 5 days in the presence of cyclopamine, a steroidal alkaloid that specifically disrupts the Hh signaling pathway, exhibit a stage-dependent inhibition of Meckel's cartilage chondroblast differentiation to mature chondrocytes. This phenotype can be rescued by exogenous FGF8, a downstream effector of Hh signaling. Taken together, our results indicate that the Hh signaling pathway is critical to Meckel's cartilage ontogenesis and the rate of chondrogenesis, but not to initial primordium formation. The reliance on Hh signaling is stage dependent. PMID- 16046862 TI - Fibrin gel-immobilized primary osteoblasts in calcium phosphate bone cement: in vivo evaluation with regard to application as injectable biological bone substitute. AB - Osteogenic injectable bone substitutes may be useful for many applications. We developed a novel injectable bone substitute based on osteoblast-fibrin glue suspension and calcium phosphate bone cement (BC). Human osteoblasts were isolated from trabecular bone samples and cultured under standard conditions. Osteoblasts were suspended in fibrinogen solution (FS). BC was cured with thrombin solution. 8 x 4 mm injectable bone discs were prepared using silicon molds and a custom-made applicator device. Discs containing BC, BC/FS, or BC/FS/osteoblasts were implanted subcutaneously into athymic nude mice. After 3, 9 and 24 weeks, specimens were explanted and subjected to morphologic and biomechanical evaluation. In vitro fibrin gel-embedded osteoblasts displayed a differentiated phenotype as evidenced by alkaline phosphatase, collagen type 1 and von Kossa stains. A proportion of osteoblasts appeared morphologically intact over a 3-day in vitro period following application into the BC. BC/FS and BC/FS/osteoblast discs were sparsely infiltrated with vascularized connective tissue. There was no bone formation in implants from all groups. However, positive von Kossa staining only in BC/FS/osteoblast groups suggests engraftment of at least some of the transplanted cells. Biomechanical evaluation demonstrated initial stability of the composites. Young's modulus and maximal load did not differ significantly in the BC/FS and BC/FS/osteoblast groups. The practicability of osteoblast-containing injectable bone could be demonstrated. The dense microstructure and the suboptimal initial vascularization of the composites may explain the lack of bone formation. Modifications with regard to enhanced osteoblast survival are mandatory for a possible application as injectable osteogenic bone replacement system. PMID- 16046863 TI - Manipulation of the phenylalanine metabolism in human keratinocytes by retroviral mediated gene transfer. AB - Phenylketonuria (PKU) is an inherited disease causing increased levels of phenylalanine in body fluids due to deficiency of hepatic phenylalanine hydroxylase (PAH) or other enzymes involved in the phenylalanine metabolism. With the long-term goal of using gene transfer to the skin to remove phenylalanine, we have previously shown that overexpression of PAH, catalyzing the hydroxylation of phenylalanine, and GTP cyclohydrolase (GTP-CH), involved in the formation of the necessary cofactor BH4,are required. Here we investigate whether manipulation of additional steps in the phenylalanine clearance pathway can further improve the phenylalanine uptake and metabolism. Transport of phenylalanine into human keratinocytes could be increased by overexpressing the two subunits LAT1 and 4F2hc of the large neutral amino acid transporter. The PAH enzyme activity was titrated by employing mutant PAH enzymes with different specific activity and by increasing the PAH copy number in transduced keratinocytes using a repeated transduction procedure. Finally, the intracellular tyrosine concentration was lowered by overexpression of tyrosinase converting tyrosine to dopaquinone. However, measured over a 24-hour period neither of these manipulations resulted in an increased phenylalanine uptake. These results suggest that other enzymes than GTP-CH, involved in BH4 synthesis and/or regeneration, can be rate-limiting in the genetically modified keratinocytes. PMID- 16046864 TI - Structure of the ovaries of the Nimba otter shrew, Micropotamogale lamottei, and the Madagascar hedgehog tenrec, Echinops telfairi. AB - The otter shrews are members of the subfamily Potamogalinae within the family Tenrecidae. No description of the ovaries of any member of this subfamily has been published previously. The lesser hedgehog tenrec, Echinops telfairi, is a member of the subfamily Tenrecinae of the same family and, although its ovaries have not been described, other members of this subfamily have been shown to have ovaries with non-antral follicles. Examination of these two species illustrated that non-antral follicles were characteristic of the ovaries of both species, as was clefting and lobulation of the ovaries. Juvenile otter shrews range from those with only small follicles in the cortex to those with 300- to 400-microm follicles similar to those seen in non-pregnant and pregnant adults. As in other species, most of the growth of the oocyte occurred when follicles had one to two layers of granulosa cells. When larger follicles became atretic in the Nimba otter shrew, hypertrophy of the theca interna produced nodules of glandular interstitial tissue. In the tenrec, the hypertrophying theca interna cells in most large follicles appeared to undergo degeneration. Both species had some follicular fluid in the intercellular spaces between the more peripheral granulosa cells. It is suggested that this fluid could aid in separation of the cumulus from the remaining granulosa at ovulation. The protruding follicles in lobules and absence of a tunica albuginea might also facilitate ovulation of non antral follicles. Ovaries with a thin-absent tunica albuginea and follicles with small-absent antra are widespread within both the Eulipotyphla and in the Afrosoricida, suggesting that such features may represent a primitive condition in ovarian development. Lobulated and deeply crypted ovaries are found in both groups but are not as common in the Eulipotyphla making inclusion of this feature as primitive more speculative. PMID- 16046865 TI - Relationship between pituitary and adipose tissue after hypothalamic denervation in the female rat. A morphometric immunohistochemical study. AB - Neonatal administration of monosodium glutamate (MSG) to rats produces severe lesions in certain hypothalamic nuclei, with repercussions in different neuroendocrine axes, and serves as a model for their study. In addition, adipose tissue, as a target organ, is known to be directly related to several neurondocrine axes. We used 21-day-old female Sprague-Dawley rats that had received a neonatal treatment with MSG (4 mg/g body weight, i.p., from day 2 up to day 10 of age) in addition to control rats (injected with 10% NaCl solution, on a similar schedule). We performed a specific immunohistochemical study on each anterior-pituitary cell population, along with the morphometry of these cells and of the parietal and visceral adipose tissue, and measured the levels of serum leptin and triglycerides. The MSG animals evinced significant changes in volume density (VD), cell density (CD), and cell size (CS) in the corticotropes, thyrotropes, and LH gonadotropes, but not in the somatotropes, lactotropes, and FSH gonadotropes. The modification common to the three cell types was a hyperplasia, but with different results depending on cell size. Furthermore, in the MSG rats significant changes were also observed in the VD, CD, and CS of the adipose tissue, consisting of adipogenesis and decrease of adipocyte size in visceral fat, together with probable lipogenesis as judged by an increase in adipocyte size in the parietal fat. The serum levels of leptin and triglycerides appeared significantly higher in MSG animals. For the first time in this animal model, and at the level of three neuroendocrine axes, our results suggest changes that correlate hypothalamic damage, cellular pituitary alterations, and the response of the adipose tissue as a target organ for MSG insult. PMID- 16046867 TI - Hormone responses to social stress in abstinent alcohol-dependent subjects and social drinkers with no history of alcohol dependence. AB - BACKGROUND: Previous studies have described blunted stress hormone responses after pharmacological activation of the hypothalamic-pituitary-adrenal (HPA) axis in sober alcoholics. The aim of the present study was to compare ACTH, cortisol, and prolactin responses to a psychological stressor in abstinent alcohol dependent subjects matched to healthy control subjects. METHODS: Individuals who met DSM-IV diagnostic criteria for a history of alcohol dependence but not for other axis I disorders were included in the study (n = 18; mean duration of abstinence +/- SEM, 3.5 +/- 5.7 years). Social drinkers (n = 23) served as control subjects. The sober alcohol-dependent and control subjects were matched for demographic measures including levels of stress symptoms. All subjects underwent the Trier Social Stress Test (TSST), a laboratory-based psychological stressor. Prestress and poststress plasma ACTH, cortisol, and prolactin levels, as well as a self-report measure of anxiety (State-Trait Anxiety Inventory), were obtained. RESULTS: Nondepressed, abstinent alcoholics and control subjects did not differ with regard to age, racial composition, or baseline or poststress ratings of anxiety. Whereas ACTH and cortisol levels increased in response to the TSST, prolactin levels did not. Stress hormone response curves for the three hormones did not differ between the alcoholics and control subjects. CONCLUSIONS: When matched for levels of stress, a laboratory-based psychological stress test did not induce differential hormone response curves for abstinent alcoholics and control subjects. PMID- 16046868 TI - Influence of age at drinking onset on long-term ethanol self-administration with deprivation and stress phases. AB - BACKGROUND: Onset of alcohol use during adolescence has potentially long-lasting consequences, e.g., prospective alcohol dependence. To obtain new insight into the effects of early chronic ethanol consumption, we compared the drinking behavior of two adult male Wistar rat groups: one that initiated alcohol consumption during adolescence (adolescent group) and the other that initiated their drinking during adulthood (adult group) in a model of long-term alcohol self-administration. We investigated the magnitude of the effects of deprivation and stress on alcohol intake and the influence of these events on the alcohol drinking behavior across time. METHODS: Heterogeneous Wistar rats aged 31 days (adolescents) and 71 days (adults) were given ad libitum access to water, as well as 5% and 20% ethanol solutions during an observation period of 30 wk. A deprivation phase of 14 days was instituted after eight wk of access to alcohol. After 16 and 26 wk of alcohol access, all animals were subjected for three consecutive days to forced swimming and electric foot shocks, respectively. RESULTS: At the onset of drinking, adolescent animals consumed less alcohol and showed lower preference than adults. The deprivation phase was followed by increased intake of highly concentrated ethanol solution without appreciable differences between age groups. Repeated swim stress produced a slight increase in ethanol consumption in both animal groups; however, alcohol intake was not significantly different between groups, whereas the foot shock stress-induced increase in alcohol intake was significantly higher in the animal group that initiated alcohol consumption during adolescence. After swim stress, the drinking behavior of the adolescent group resembled that of the adult group. In particular, the adolescent group increased their preference for 20% ethanol solution for the remainder of the experiment. CONCLUSIONS: Age of voluntary alcohol drinking onset does not appear to be a strong predictor for prospective alcohol intake and relapse-like drinking behavior under the present experimental conditions. However, male Wistar rats that initiated alcohol consumption during adolescence seem to be more susceptible to acute stressor-specific effects in terms of alcohol consumption. PMID- 16046869 TI - Manipulations of serotonin function in the nucleus accumbens core produce differential effects on ethanol and sucrose seeking and intake. AB - OBJECTIVE: Behaviorally relevant stimuli, including alcohol, are processed through the nucleus accumbens/ventral tegmental area (VTA)/prefrontal cortex circuit. It is hypothesized that serotonin affects ethanol-directed behaviors by interacting with this system via projections from the dorsal raphe to the nucleus accumbens and VTA. The current studies utilized two different operant paradigms, one focusing on reinforcer seeking and one focusing on reinforcer self administration (both with an ethanol and a sucrose solution as the reinforcer) to elucidate serotonin-specific regulation of these behaviors. METHODS: The present experiments assessed the effects of microinjections of a serotonin1B agonist (CGS12066B) and a serotonin1A agonist (8-OH-DPAT) in the nucleus accumbens core on ethanol- and sucrose-reinforced seeking and intake. In four separate experiments, male Long-Evans rats were trained to complete a single response requirement that resulted in access to 10% ethanol or 2% sucrose for a 20-min drinking period. RESULTS: Before microinjections, ethanol-reinforced subjects were consuming an average of 0.5-0.95 g/kg ethanol and making 50-100 responses during intermittent nonreinforced sham (no drug) sessions (sucrose groups had similar baseline response levels). In summary, findings from the four experiments showed the following: (1) manipulations of serotonin function that had effects on ethanol-reinforced responding had either no effect or less pronounced effects on sucrose-reinforced responding; (2) administration of the serotonin1B agonist decreased seeking behaviors to a greater degree than drinking behaviors; and (3) administration of the serotonin1A agonist decreased ethanol intake but not seeking with no impact at all on sucrose-reinforced behaviors. CONCLUSIONS: Manipulations of serotonin activity in the nucleus accumbens core had little effect on sucrose-reinforced behaviors and differential effects on ethanol seeking versus intake, suggesting that this area may play a complex but selective role in the stimulus processing of external and internal alcohol-associated cues. PMID- 16046870 TI - Prevention of ethanol-induced behavioral stimulation by D-penicillamine: a sequestration agent for acetaldehyde. AB - BACKGROUND: D-Penicillamine, a sulfhydryl amino acid derived from penicillin, is a highly selective agent for sequestering in vivo acetaldehyde, the first metabolic product of ethanol. A substantial amount of research supports the idea that brain acetaldehyde, produced by central ethanol metabolism, plays a key role in determining some of the behavioral effects of ethanol administration. This study addressed two questions. First, we tested if D-penicillamine was able to modify the depressant effects of acetaldehyde on behavior. Second, we studied the effect of D-penicillamine on ethanol-induced behavioral stimulation. METHODS: Mice were pretreated with 75.00 mg/kg of D-penicillamine, and 30 min later, they received acetaldehyde at 0, 100, 200, or 300 mg/kg intraperitoneally. Different groups of animals were treated with 0.0, 37.5, 75, 150, or 300 mg/kg of D penicillamine simultaneously 30, 90, 150, or 210 min before the intraperitoneal administration of saline or 1.2, 1.8, 2.4, 3.0, or 3.6 g/kg of ethanol, respectively. The specificity of D-penicillamine effects was addressed using two drugs: cocaine (4 mg/kg) and caffeine (15 mg/kg). RESULTS: Our results revealed that behavioral depression caused by acetaldehyde (200 and 300 mg/kg) could be attenuated by D-penicillamine treatment. In addition, D-penicillamine was also effective in lowering behavioral locomotion induced by ethanol (1.8 and 2.4 g/kg), without altering spontaneous locomotor activity. This sulfhydryl amino acid specifically modified the effect of ethanol on locomotion because cocaine- or caffeine-induced locomotion was unaffected. In addition, blood ethanol levels were not different between D-penicillamine- and saline-pretreated mice. CONCLUSIONS: Behavioral effects produced by acetaldehyde and ethanol are blocked when animals are treated with D-penicillamine, an effective sequestration agent for acetaldehyde. These results suggest that some of the psychopharmacological effects, classically attributed to ethanol, could be mediated by its first metabolite, acetaldehyde. PMID- 16046871 TI - Hangover susceptibility in relation to aldehyde dehydrogenase-2 genotype, alcohol flushing, and mean corpuscular volume in Japanese workers. AB - BACKGROUND: A study of Asian-American students suggested a positive association between inactive ALDH2*2 and susceptibility to hangover. A biomarker for moderate to-heavy drinking in persons with inactive aldehyde dehydrogenase-2 (ALDH2) is increased mean corpuscular volume (MCV). METHODS: Associations between hangover and ALDH2 genotype, alcohol flushing, and MCV were examined for 251 Japanese workers (139 men, 112 women). RESULTS: Inactive ALDH2*1/2*2 heterozygotes drank less alcohol than active ALDH2*1/2*1 homozygotes (p < 0.0001), but the frequency of hangover did not significantly differ between the two groups for either gender. The amount of drinking reported to lead to hangover was significantly less for male and female ALDH2*1/2*2 heterozygotes than for their ALDH2*1/2*1 homozygous counterparts (p < 0.005). The proportion of men who had hangover three times or more during the past year increased significantly with increased daily alcohol consumption in men with the ALDH2*1/2*2 genotype (p = 0.0002) but not in those with the ALDH2*1/2*1 genotype. For men who usually consumed <44 g of ethanol/day, the median amount of drinking before hangover was significantly lower for ALDH2*1/2*2 men than for ALDH2*1/2*1 men reporting the same level of consumption. Hangover occurred with consistently high frequency among ALDH2*1/2*1 men, regardless of their daily consumption. Similar findings were observed in a comparison of men who never flushed and those who reported current or former flushing, a surrogate marker of inactive ALDH2. Assessment of hangover risk by quartiles of MCV showed that men with MCV of > or =96 had a significantly higher risk of hangover than did men with MCV of <91 (odds ratio = 5.56; 95% confidence interval = 1.69-18.25). CONCLUSIONS: Inactive heterozygous ALDH2, alcohol flushing, and increased MCV were positively associated with hangover susceptibility in Japanese workers, suggesting that acetaldehyde is etiologically linked to the development of hangover. PMID- 16046872 TI - Development of a chinese version of the Yale-Brown obsessive compulsive scale for heavy drinking. AB - BACKGROUND: Measurement of craving is an important component in the investigation of the etiology and clinical pictures of alcoholism and dependence of other substances in different cultures. The aim of this study was to develop a Chinese version of the Yale-Brown Obsessive Compulsive Scale for heavy drinking (YBOCS-hd C), the instrument most frequently used in assessing the severity of alcohol craving in Taiwan. METHODS: Four hundred twenty Han Chinese (220 with alcohol use disorders) and 218 Bunun aborigines (150 with alcohol use disorders) in Taiwan were interviewed by mental health professionals with the YBOCS-hd-C and a Chinese version of the World Health Organization Schedules for Clinical Assessment in Neuropsychiatry to establish the psychiatric diagnosis. Validity and reliability of the YBOCS-hd-C were examined. RESULTS: The YBOCS-hd-C was found to have acceptable interrater reliability (intraclass correlation, 0.89-0.96), internal consistency (Cronbach's alpha = 0.99), construct validity, concurrent validity, and cross-cultural validity. The correlations between 10 items of the YBOCS-hd-C and 11 items of the World Health Organization Schedules for Clinical Assessment in Neuropsychiatry adjusted for age, gender, and ethnicity ranged from 0.39 to 1.00. The YBOCS-hd-C also discriminated effectively among individuals with alcohol dependence, alcohol abusers, and normal drinkers. CONCLUSIONS: This study demonstrated that the YBOCS-hd-C is a reliable and valid instrument for assessing the extent of craving for alcohol in Taiwanese Han and Bunun individuals. PMID- 16046873 TI - Toward efficient and comprehensive measurement of the alcohol problems continuum in college students: the brief young adult alcohol consequences questionnaire. AB - BACKGROUND: Although a number of measures of alcohol problems in college students have been studied, the psychometric development and validation of these scales have been limited, for the most part, to methods based on classical test theory. In this study, we conducted analyses based on item response theory to select a set of items for measuring the alcohol problem severity continuum in college students that balances comprehensiveness and efficiency and is free from significant gender bias. METHOD: We conducted Rasch model analyses of responses to the 48-item Young Adult Alcohol Consequences Questionnaire by 164 male and 176 female college students who drank on at least a weekly basis. An iterative process using item fit statistics, item severities, item discrimination parameters, model residuals, and analysis of differential item functioning by gender was used to pare the items down to those that best fit a Rasch model and that were most efficient in discriminating among levels of alcohol problems in the sample. RESULTS: The process of iterative Rasch model analyses resulted in a final 24-item scale with the data fitting the unidimensional Rasch model very well. The scale showed excellent distributional properties, had items adequately matched to the severity of alcohol problems in the sample, covered a full range of problem severity, and appeared highly efficient in retaining all of the meaningful variance captured by the original set of 48 items. CONCLUSIONS: The use of Rasch model analyses to inform item selection produced a final scale that, in both its comprehensiveness and its efficiency, should be a useful tool for researchers studying alcohol problems in college students. To aid interpretation of raw scores, examples of the types of alcohol problems that are likely to be experienced across a range of selected scores are provided. PMID- 16046874 TI - A temporal and dose-response association between alcohol consumption and medication adherence among veterans in care. AB - BACKGROUND: Previous studies have shown that alcohol consumption is associated with decreased medication adherence, but this association may be confounded by characteristics common among those who drink heavily and those who fail to adhere (e.g., illicit drug use). Our objective was to determine whether there are temporal and dose-response relationships between alcohol consumption and poor adherence. METHODS: We administered telephone interview surveys to participants in the Veterans Aging Cohort Study, an eight-site observational study of HIV+ and matched HIV- veterans in care, to determine whether alcohol consumption on a particular day was associated with nonadherence to prescribed medications on that same day. We used the Time Line Follow Back to measure alcohol consumption and the Time Line Follow Back Modified for Adherence to measure adherence. Individuals were categorized as abstainers (no alcohol in past 30 days), nonbinge drinkers (alcohol in past 30 days but < or =four standard drinks on each day), or binge drinkers (> or =five standard drinks on at least one day). RESULTS: Among 2702 respondents, 1582 (56.6%) were abstainers, 931 (34.5%) were nonbinge drinkers, and 239 (8.9%) were binge drinkers. Abstainers missed medication doses on 2.4% of surveyed days. Nonbinge drinkers missed doses on 3.5% of drinking days, 3.1% of postdrinking days, and 2.1% of nondrinking days (p < 0.001 for trend), and this trend was more pronounced among HIV+ individuals than HIV- individuals. Binge drinkers missed doses on 11.0% of drinking days, 7.0% of postdrinking days, and 4.1% of nondrinking days (p < 0.001 for trend), and this trend was comparably strong for HIV+ and HIV- individuals. CONCLUSIONS: Among veterans in care, self-reported alcohol consumption demonstrates a temporal and dose-response relationship to poor adherence. HIV+ individuals may be particularly sensitive to alcohol consumption. PMID- 16046875 TI - Effects of systemic and local CXC chemokine administration on the ethanol-induced suppression of pulmonary neutrophil recruitment. AB - BACKGROUND: Acute alcohol intoxication impairs the neutrophil response to intrapulmonary infection, resulting in impaired host defense and increased patient morbidity and mortality. We recently showed that intratracheal (IT) chemokine administration promotes pulmonary neutrophil migration in rats and that this process is enhanced by systemic administration of the Glu-Leu-Arg (ELR+) and CXC chemokine cytokine-induced neutrophil chemoattractant (CINC). Here we hypothesized that exogenous chemokine administration would mitigate the suppressive effect of alcohol on neutrophil recruitment into the lung. METHODS: Macrophage inflammatory protein-2 (MIP-2), a rat ELR+ CXC chemokine, or live Klebsiella pneumoniae (K. pneumoniae) was administered it to induce alveolar neutrophil migration in the absence or presence of acute ethanol intoxication. Depending on the experimental protocol, rats received either intravenous (IV) CINC or IT chemokines (CINC and MIP-2) 20 min after it MIP-2 or K. pneumoniae. Rats were euthanized 90 min or four hr after the first IT injection for sample collection. RESULTS: Neutrophil counts were significantly elevated in bronchoalveolar lavage fluid (BALF) of rats receiving IT MIP-2 compared with vehicle-treated rats, and this response was significantly decreased in animals pretreated with ethanol. CINC IV enhanced the neutrophil response to IT MIP-2 in both the absence and presence of acute ethanol intoxication. In rats challenged with K. pneumoniae, ethanol pretreatment significantly reduced BALF levels of CINC and MIP-2, suppressed alveolar neutrophil recruitment, and decreased whole lung myeloperoxidase activity. CINC IV did not alter BALF neutrophil counts in the absence or presence of ethanol administration 4 hr after IT K. pneumoniae. Alternatively, IT chemokine instillation partially restored BALF neutrophil recruitment but not whole-lung myeloperoxidase activity in ethanol-treated rats. CONCLUSIONS: Ethanol significantly inhibits the pulmonary inflammatory responses to both MIP-2 and K. pneumoniae. Exogenous chemokine administration may be a useful means to enhance host defenses in the ethanol-intoxicated host, although the results of this study also indicate that ethanol intoxication can impair neutrophil recruitment, independent of its effects on local chemotactic gradients. PMID- 16046876 TI - The -159C/T polymorphism in the promoter region of the CD14 gene is associated with advanced liver disease and higher serum levels of acute-phase proteins in heavy drinkers. AB - BACKGROUND: Innate inflammatory responses to endotoxin (lipopolysaccharide) contribute to the development of alcoholic liver disease (ALD). A single nucleotide polymorphism (-159C/T) in the promoter region of the gene coding for CD14 (a lipopolysaccharide receptor) could be associated with the development of ALD. We sought too investigate the relationship between the CD14/-159C/T polymorphism and advanced ALD and acute-phase protein levels in heavy drinkers. METHODS: A total of 138 heavy drinkers consecutively admitted to an Internal Medicine department were genotyped for the CD14/-159C/T polymorphism. Serum samples were analyzed for lipopolysaccharide-binding protein (LBP), soluble CD14 (sCD14), C-reactive protein (CRP), and immunoglobulin (Ig) A, IgG, and IgM. Patients with ascites or liver encephalopathy (n = 35) were classified as having advanced ALD. RESULTS: After adjusting for potential confounding variables, the CD14/-159TT genotype was positively associated with advanced ALD (odds ratio, 2.99; 95% confidence interval, 1.09-8.24, p = 0.03) and serum LBP (p = 0.01) and sCD14 (p = 0.04) levels. The CD14/-159C/T polymorphism was not associated with serum levels of CRP, IgA, IgG, or IgM. CONCLUSIONS: Our results support the notion that CD14/-159TT homozygous heavy drinkers have higher levels of the LPS binding acute-phase proteins (LBP and sCD14) than do carriers of the CD14/-159C allele. Also, the CD14/-159TT genotype may be a risk factor for advanced ALD. PMID- 16046877 TI - Evaluation of corpus callosum anisotropy in young adults with fetal alcohol syndrome according to diffusion tensor imaging. AB - BACKGROUND: Fetal alcohol syndrome (FAS) and associated disorders resulting from maternal alcohol use during gestation are among the most common developmental disorders. However, they are rarely diagnosed and not fully understood in terms of their behavioral and neurocognitive phenotype. Prenatal exposure leads to alterations in facial morphology, growth, and neurocognition. The nature and extent of teratogenic effects on the brain and the relationship between such effects and observed behaviors remain in debate because there are no established markers for the neurological effects of exposure. In this study, we examined the impact of prenatal alcohol exposure on white-matter integrity in the corpus callosum by using diffusion tensor imaging (DTI) and herein describe the relationship between such effects and observed physical and behavioral outcomes. METHODS: DTI was used to evaluate diffusion anisotropy in the genu and splenium of corpus callosum in 16 low-income, primarily African-American volunteers. Volunteers were recruited from a cohort of young adults who had received neuropsychological evaluations during adolescence. Nine had been prenatally exposed to alcohol and had characteristics of FAS, and seven were nonexposed controls. RESULTS: Significant difference in the means for diffusion fractional anisotropy (t = 2.26, df = 9, p <0.002) and apparent diffusion coefficient (t = 2.14, df = 14, p < 0.008) were observed in the corpus callosum of alcohol-exposed youth compared with nonexposed youth. No significant differences were found in intracranial volume between these groups. CONCLUSIONS: Our results illustrate that DTI can be used in evaluating the integrity of corpus callosum in alcohol exposed individuals. If future studies support these findings, diffusion anisotropy, represented by fractional anisotropy, has the potential to be used as a clinical marker in the diagnosis of FAS. PMID- 16046878 TI - Antioxidant pretreatment does not ameliorate alcohol-induced Purkinje cell loss in the developing rat cerebellum. AB - BACKGROUND: Recent research has suggested that oxidative stress is a potential mechanism for alcohol-induced injury and that supplementation with antioxidants can ameliorate alcohol-induced damage. In this study, two known antioxidants, melatonin and U83836E, were assessed for their effectiveness in blocking the expected alcohol-induced cerebellar Purkinje cell loss in neonatal rat pups. METHODS: Sprague-Dawley rat pups were artificially reared from postnatal days (PDs) 4-9 and were exposed to either alcohol or antioxidants (melatonin or U83836E) individually or in combination. A normal control group (raised by rat dams) was included in this study. On PD 9, the brain from each pup was removed and weighed, and the cerebellar vermis was processed for stereological cell counting. RESULTS: Alcohol exposure during the brain growth spurt produced microencephaly, in addition to significant decreases in the number and density of Purkinje cells in lobule I and the volume of lobule I. The antioxidants did not reduce any of the adverse effects observed from alcohol exposure, and they did not decrease the Purkinje cell number when administered alone. Furthermore, antioxidants did not change the only blood alcohol concentration measured on PD 6. CONCLUSIONS: The results confirmed alcohol-induced microencephaly and cerebellar Purkinje cell loss from neonatal alcohol exposure, and they showed that neither antioxidant could attenuate these adverse effects on the developing brain. The inability of antioxidants to reduce Purkinje cell loss from neonatal alcohol exposure suggests the existence of alternative mechanisms for developmental alcohol-induced Purkinje cell loss. PMID- 16046879 TI - Alcohol consumption, lung function, and quality of life in pneumoconiosis. AB - BACKGROUND: To our knowledge, there are no previous data on drinking problems in patients with pneumoconiosis. The aim of this study was to examine drinking patterns and the impact of drinking on lung function and health-related quality of life (HRQOL) in Chinese patients with pneumoconiosis. METHODS: Three hundred patients with pneumoconiosis were recruited from a community-based case registry. The HRQOL was measured with the St. George's Respiratory Questionnaire (SGRQ). Pulmonary function, comorbidity, and psychosocial variables were also assessed. The alcohol use of the patients was evaluated with the Alcohol Use Disorders Identification questionnaire. RESULTS: Of the 300 patients, 72.3% (217) reported that they had not consumed any alcohol during the past year, whereas 83 patients (27.7%) did report drinking alcohol. In the drinking group, 88.0% (73) consumed <7 standard drinks per week, and none of them exceeded the safety limit of 21 standard drinks per week. The drinking group (n = 83) was younger, had less concurrent medical diseases, and lower (i.e., better) unadjusted SGRQ symptom, activity, impact, and total scores than the nondrinking group (n = 217). The SGRQ scores, which were adjusted for age, duration of occupation, concurrent medical diseases, smoking status, and forced expiratory volume in 1 sec predicted tests (FEV1%), remained significantly lower for the drinking group. Although the drinking group had a higher unadjusted FEV1% predicted, the difference between the FEV1% of the two groups, after adjustment for covariates, was of borderline significance only. CONCLUSIONS: Most Chinese patients with pneumoconiosis in this study did not consume alcohol, and among those who did, the level of alcohol consumption was low. This low level of alcohol consumption was associated with a better HRQOL and possibly with better lung function. PMID- 16046880 TI - Ethanol triggers neural crest apoptosis through the selective activation of a pertussis toxin-sensitive G protein and a phospholipase Cbeta-dependent Ca2+ transient. AB - BACKGROUND: Alcohol is a potent neurotoxin that triggers the selective apoptosis of neuronal populations in the developing fetus. For neural crest cells, clinically relevant ethanol levels (0.3%) rapidly elicit a phospholipase C (PLC) dependent intracellular Ca2+ transient that is sufficient to activate apoptosis. We investigated the biochemical origins of this Ca2+ transient. METHODS: Three somite chick embryos (stage 8-) were pretreated with agonists and antagonists of PLC signaling pathways before ethanol challenge. The resulting intracellular Ca2+ release was quantified using Fluo-3; apoptosis was assessed using vital dyes. RESULTS: Pretreatment of embryos with PLC antagonists U73122 or ET-18-OCH3 confirmed that a phosphoinositide-specific PLC was required for both the ethanol dependent Ca2+ transient and subsequent cell death. Ethanol rapidly elevated intracellular inositol-1,4,5-trisphosphate [Ins(1,4,5)P3] levels in the rostral portion of the embryo that contains neural crest progenitors. The Ins(1,4,5)P3 receptor antagonist xestospongin C blocked the appearance of the ethanol dependent Ca2+ transient. Pretreatment with the pan-Galpha protein antagonist GDPbetaS, but not with the tyrosine kinase antagonist genistein, suppressed ethanol's ability to elicit the Ca2+ transient, suggesting that a rise in PLC activity and Ins(1,4,5)P3 concentration originates from stimulation of heterotrimeric G proteins. To probe the identity of this G protein, embryos were treated with G protein antagonists. Pertussis toxin and NF023 suppressed the ethanol-induced Ca2+ transient and subsequent neural crest apoptosis, whereas suramin was weakly inhibitory. C3 exoenzyme was embryolethal over a wide concentration range, consistent with suggestions that Rho family GTPases participate in neural crest development. Galphai2 was identified by immunostaining in the neural crest cells. CONCLUSION: We propose a role for Galphai/o protein activation and subsequent interaction of Gbetagamma with PLCbeta in mediating the proapoptotic effects of ethanol upon the developing neural crest. PMID- 16046881 TI - Temporal differences in the ability of ethanol to modulate endotoxin-induced increases in inflammatory cytokines in muscle under in vivo conditions. AB - BACKGROUND: Acute alcohol (EtOH) intoxication may both antagonize and potentiate the ability of monocytes/macrophages to respond to endotoxin (lipopolysaccharide [LPS]). The suppressive effects of EtOH predominate when the duration between EtOH and LPS administration is relatively short, whereas sensitization is observed under conditions when there is a relatively longer delay between EtOH and LPS exposure. Striated muscle is now recognized to possess components of both the afferent and efferent limbs of the innate immune system. The aim of the present study was to determine whether the interval between EtOH and LPS administration differentially affects the mRNA content for selected elements of the innate immune response in skeletal and cardiac muscle and to compare such changes with those occurring in liver and spleen. METHODS: The content of mRNA for interleukin (IL)-6, IL-1beta, tumor necrosis factor (TNF)-alpha, and high mobility group box (HMGB)-1, as well as toll-like receptors (TLRs)-2 and -4, were measured in gastrocnemius, heart, liver and spleen from rats orally gavaged with EtOH and then injected with LPS either two or 24 hr thereafter. RESULTS: EtOH intoxication two hr before LPS acutely suppressed the increased IL-6 mRNA in all tissues and antagonized the increase in plasma and tissue IL-6 protein concentration. Similarly, EtOH blunted the LPS-induced increase in tissue mRNA expression of TNF-alpha and IL-1beta. In contrast, when LPS was given 24 hr after EtOH, the increased IL-6 in striated muscle, but not in liver or spleen, was selectively potentiated. An enhanced LPS responsiveness was also observed for the late-phase cytokine HMGB1 in all tissues; however, the increased tissue expression of TNF-alpha and IL-1beta induced by LPS was not augmented. TLR4 mRNA was decreased in both heart and spleen (but unaltered in skeletal muscle and liver) of rats injected with LPS, and this change was prevented by pretreatment with EtOH. In contrast, EtOH alone increased TLR-2 mRNA content of heart, liver, and spleen but not muscle. LPS also markedly increased TLR2 mRNA in the same three tissues under control conditions, but this increase was attenuated by EtOH administered either two or 24 hr before LPS. CONCLUSIONS: Under in vivo conditions, the interval between EtOH exposure and LPS differentially affected the synthesis of various cytokines. In this regard, EtOH administered within two hr of LPS generally suppressed IL-6, IL-1beta, and TNF-alpha mRNAs in muscle, heart, liver, and spleen. Delaying the exposure of animals to LPS for 24 hr after EtOH, however, accentuated the increase in IL-6 and HMGB1, and for IL-6, this increased sensitivity appeared localized to striated muscle. PMID- 16046882 TI - Single-amino-acid difference in the sequence of alpha6 subunit dramatically increases the ethanol sensitivity of recombinant GABAA receptors. PMID- 16046885 TI - Chemotherapy-induced nausea/vomiting and functional status in women treated for breast cancer. AB - Nausea and vomiting are among the most distressing symptoms for cancer patients treated with chemotherapy even with the widespread use of 5-HT3 antagonists. Chemotherapy-induced nausea and vomiting (CINV) is composed of 4 major components: acute nausea, delayed nausea, acute vomiting, and delayed vomiting. Determining the relationship of each component of CINV on the functional status of women undergoing chemotherapy for breast cancer was the purpose of this study. This longitudinal, descriptive study recruited 303 patients with breast cancer undergoing chemotherapy from 40 study sites in the United States. Reliable and valid measures of CINV and functional status were employed. Patients demonstrated significant decreases in the following aspects of functional status as measured by the SF-36: physical functioning (P < .0005), role limitations due to physical problems (P = .003), general health (P = .029), vitality (P < .0005), and social functioning (P = .001). The pattern of reduction in usual activities and increase in hours of resting correlated best with 2 components of CINV--delayed nausea and delayed vomiting (P < .0001, each). The results of this study suggest that control of delayed CINV may contribute to the functional improvement of women receiving chemotherapy for breast cancer. PMID- 16046886 TI - Testicular cancer and testicular self-examination: knowledge and attitudes of adolescent Swedish men. AB - The main objective of this study was to assess the knowledge of and attitudes toward testicular cancer (TC) and the prevalence of testicular self-examination (TSE) among senior high school male students in Sweden. Another aim was to describe to what extent the students had received information about TC and TSE prior to this study. Seven hundred twenty-seven (70.7%) male students (M = 17 years) completed a 60-item questionnaire, encompassing background characteristics, information, knowledge of and attitudes toward TC, and prevalence of TSE practice. The results showed that most students had never heard of TC or TSE, had limited knowledge of common symptoms, and had hardly ever practiced TSE. The most frequently reported information source on this topic was the mass media. Most of the students considered TC to be a serious disease and were afraid of getting it. Thus, there is a need for imparting health education with respect to this subject, information that could preferably be included when promoting health in school settings and during enlistment to the military services. This investigation is the first part of an intervention study aiming to describe the effect of different educational materials on male students' knowledge of TC and their practice of TSE. PMID- 16046887 TI - Symptom distress changes during first postoperative month in newly diagnosed Taiwanese breast cancer patients: a longitudinal study. AB - The purpose of this longitudinal study was to explore changes in symptom distress in newly diagnosed Taiwanese breast cancer patients during the initial 4-week postoperative period. The research instruments, including a demographic questionnaire and the Symptom Distress Scale, were used to obtain data on postoperative day 2 and at weeks 2, 3, and 4. In total, 39 patients with a mean age of 48 years participated in this study. Data were analyzed using descriptive statistics, t tests, one-way ANOVA, and repeated-measures ANOVA. Results revealed that the level of symptom distress significantly decreased from postoperative day 2 to week 4. Loss of appetite and a poor outlook increased; nausea frequency, fatigue, and insomnia decreased then increased; and frequency and the level of pain, coughing, tightness/tenderness in the chest wall, weakness, and numbness in the arm of the operative side all decreased over the 4-week study period. Age, stage of disease, and type of surgery were all related to symptom distress. Results of this study may provide reassurances about what can be expected after breast cancer surgery. PMID- 16046888 TI - Symptom clusters: concept analysis and clinical implications for cancer nursing. AB - The purpose of this article is to analyze the concept of symptom clusters and to discuss its application to cancer nursing to promote communication and enhance scientific knowledge. Rodgers' evolutionary method of concept analysis served as the framework for reviewing literature from psychology/psychiatry, general medicine, and nursing. Attributes of symptom clusters were relationships of symptoms and relationships of clusters, concurrence, underlying dimensions, stability, and common etiology. The major antecedent was the presence of 2 or more symptoms. Consequences were poorer physical health status, interference with activities of daily living, emotional distress, and increased financial burden. A symptom cluster is defined as consisting of 2 or more symptoms that are related to each other and that occur together. Symptom clusters are composed of stable groups of symptoms, are relatively independent of other clusters, and may reveal specific underlying dimensions of symptoms. Relationships among symptoms within a cluster should be stronger than relationships among symptoms across different clusters. Symptoms in a cluster may or may not share the same etiology. Symptom should be broadened to include both subjective (self-reported) symptoms and objective (observed) signs. Implications for researchers include the need to use a clear definition, determine the optimal methods of identifying etiology and nature of symptom clusters in various populations, assess the clinical utility of symptom clusters, and test interventions. Implications for practitioners include the need to comprehensively assess symptoms over the entire cancer trajectory, select interventions that target single and multiple symptoms, and evaluate outcomes that include quality of life and economic variables. PMID- 16046891 TI - Spirit body healing--a hermeneutic, phenomenological study examining the lived experience of art and healing. AB - Spirit-Body Healing was a hermeneutic, phenomenological research study that examined the lived experience of art and healing (using Max Van Manen's method of researching the lived experience) of 63 participants over a 4-year period. Data included taped interviews with artists and hospitalized cancer patients who could articulate their lived experience of art and healing, observations of artists at work, journal writings, and material from the Arts In Medicine program at Shands Hospital, University of Florida. The researcher interpreted data through Van Manen's method of writing as inquiry to elucidate the themes of the participants' experiences. The themes that emerged were (1) going into the darkness, (2) going elsewhere, (3) making art as a turning point, (4) slipping through the veil, (5) feeling the healing energy of love and compassion, (6) surrendering to the process, (7) knowing the truth, and (8) experiencing transcendence. Understanding the power of art to heal mind, body, and spirit has important implications for cancer nursing practice, and nurses can provide the leadership and commitment to integrate the arts into patient care. PMID- 16046892 TI - More than trivial: strategies for using humor in palliative care. AB - Humor and laughter are ubiquitous in human interactions. Terminal illness, however, is often accompanied by circumstances of anxiety, fear, and sadness. Hospice/palliative care emphasizes quality of life and the importance of human relationships. In this context, humor finds its place in authentic person-to person connectedness. This article presents findings from a clinical ethnography that investigated the phenomena of humor and laughter in an inpatient palliative care unit. As a participant observer, the lead author accompanied 6 nurses throughout their day-to-day activities, twice weekly over 12 weeks. In addition to more than 200 hours of fieldwork, informal conversations were held with patients and families and semistructured interviews were conducted with nurses (n = 11), physicians (n = 2), a social worker (n = 1), and a physiotherapist (n = 1). Humor was pervasive, varied in the setting, and occurred across a range of intensities. Both clients and team members used humor to build relationships, contend with circumstances, and express sensibilities. Humor was affected by differences in people, differing circumstances, ethnicity, gender, and degree of stress. Participants relied on intuition as well as a constellation of other factors in discerning whether or not to use humor. Techniques for assessment included identification of cues such as expression in the eyes and timing as indications of receptivity. Combined with caring and sensitivity, humor is a powerful therapeutic asset in hospice/palliative care. It must neither be taken for granted nor considered trivial. PMID- 16046894 TI - Dance and movement program improves quality-of-life measures in breast cancer survivors. AB - A pilot research study was conducted at 2 cancer centers in Connecticut to determine the effect of a dance and movement program on quality of life and shoulder function in breast cancer survivors treated within the prior 5 years. Thirty-five women completed the trial that included a 12-week intervention, using The Lebed Method, Focus on Healing Through Movement and Dance. The study design was a randomized control trial with a wait list control group crossover to active treatment in weeks 13 to 25, with the treatment group receiving the program in weeks 1 to 12, and no program in weeks 13 to 25. Outcome measures were the Breast Cancer Quality of Life (FACT-B), Shoulder range of motion (ROM), and Body Image Scale. FACT-B significantly improved in the intervention group at 13 weeks from 102.0 +/- 15.8 to 116.7 +/- 16.9, compared to the wait list group 108.1 +/- 16.4 to 107.1 +/- 21.3 (time x group effect, P = .008). During the crossover phase, the FACT-B score increased in the wait list group and was stable in the treatment group. The overall effect of the training at 26 weeks was significant (time effect, P = .03), and the order of training was also significant (P = .015). Shoulder ROM increased in both groups at 13 weeks--15 degrees and 8 degrees in the intervention and wait list groups (Time effect, P = .03; time x group, P = .58). Body Image improved similarly in both groups at 13 weeks (time effect, P = .001; time x group, P = .25), and at 26 weeks. There was no significant effect of the order of training for these outcome measures. A dance movement program that addressed the physical and emotional needs of women following treatment for breast cancer substantially improved a breast cancer-specific quality-of-life measure. Larger studies are justified to determine the acceptability of this therapy as part of the continuum of care for breast cancer survivors. PMID- 16046895 TI - Living after external beam radiotherapy of localized prostate cancer: a qualitative analysis of patient narratives. AB - The treatment of prostate cancer induces adverse effects. Although quantitative studies have evaluated the influence of these adverse effects on the quality of life, few studies have tried to gain a deeper understanding of how men live after external beam radiotherapy of localized prostate carcinoma, which is the purpose of this study. Ten men were interviewed in their homes. The narrative interviews were tape recorded and transcribed into a text. The text was analyzed qualitatively by a content analysis. To bear the emotional experience of the illness by oneself is a self-chosen strategy. Some men expressed a sense of being exposed in meetings with female caregivers. The treatment induced changes in body functions influencing daily life. In this new life situation these men are striving to reach a sense of having control, which includes control over disease progression and waning body function. Despite the negative influence of the treatment, the men are striving to become reconciled with their new life. The experience of living after external beam radiotherapy of localized prostate carcinoma could be understood as striving to reach a sense of control and becoming reconciled with a new way of life. PMID- 16046896 TI - Pain and desire for hastened death in terminally ill cancer patients. AB - The purpose of this study was to assess the relationship between pain and the desire for hastened death in terminally ill cancer patients. The participants were 120 terminally ill cancer patients under palliative treatment from June 2003 to November 2004. Patients completed a pain assessment tool, the Greek Brief Pain Inventory (G-BPI), and a self-report measure of the desire for hastened death, the Greek Schedule of Attitudes Toward Hastened Death (G-SAHD). Moderate but statistically significant associations were found between some of the severity and interference items of G-BPI and G-SAHD; more specifically, between G-SAHD and G-BPI3, "worst pain in the last 24 hours" (r = 0.279, P = .002); G-SAHD and G BPI4, "least pain in the last 24 hours" (r = 0.253, P = .005); and G-SAHD and G BPI5, "average pain in the last 24 hours" (r = 0.283, P = .002). A stronger association was revealed between G-SAHD and G-BPI8, "relief provided by pain treatment and medications in the last 24 hours" (r = -0.326, P = .000). Multiple regression analyses including the enter model and the forward model were conducted. According to the enter model, the strongest predictors of hastened death were items G-BPI6, "current pain"; G-BPI8, "relief provided by pain treatment and medications in the last 24 hours"; G-BPI9i, "interference of pain in general activity"; and G-BPI9iii, "interference of pain in walking." According to the forward model, significant predictors of the desire for death were items G BPI5, "average pain in the last 24 hours"; G-BPI6, "current pain"; G-BPI9i, "interference of pain in general activity"; and G-BPI9ii, "interference of pain in mood," all of which were statistically significant (P = .000-.042). Pain appeared to have a statistically significant relationship with the desire for hastened death. Effective treatment by healthcare professionals should be provided to reduce pain and cancer-related symptoms as well as the desire for hastened death. PMID- 16046897 TI - Depressive symptoms prescreening and postscreening among returning participants in an ovarian cancer early detection program. AB - Ovarian cancer is the fourth leading cause of cancer mortality among women. Previous research has shown that initial ovarian cancer screening has the potential to cause depressive symptoms among women at increased risk for the disease but no study has evaluated depressive symptoms shortly after screening. This article explores depressive symptoms prescreening and postscreening in women returning to participate in an ovarian cancer early detection program. Seventy two women, with a mean age of 48, most with a family history of ovarian cancer and/or a personal history of breast cancer, completed the Center for Epidemiologic Studies Depression Scale (CES-D) immediately prior to screening and 1 week following. CES-D scores at prescreening (13%) were significantly lower than previously reported. No statistical differences in CES-D scores before and after screening were found, although 75% of women with elevated pre-CES-D scores had scores below the cutoff at postscreening. Although our numbers are lower than reported in previous studies, they are important because they emphasize the need for continued assessment of individuals who may be at greater risk for psychological distress related to cancer screening. Internationally, nurses play an important role in the recognition and support of patients undergoing cancer screening. PMID- 16046899 TI - New insights in the pharmacological therapy of arterial hypertension. AB - PURPOSE OF REVIEW: Despite the large number of antihypertensive drugs available, their usefulness is limited due to low efficacy, side effects, poor patient compliance and failure to reduce the cardiovascular risk to the level of the general population. These limitations have stimulated the research and development of new antihypertensive drugs, which we briefly review herein. RECENT FINDINGS: Novel antihypertensive drugs under development include new oral renin inhibitors, brain aminopeptidase A inhibitors, angiotensin-converting enzyme 2 modulators, short-acting L-type Ca channel blockers, new T-type Ca channel blockers, inhibitors of vascular NAD(P)H oxidase, angiotensin-converting enzyme crosslink breakers, Rho kinase inhibitors, renal Na/K ATPase inhibitors, potassium channel openers and drugs combining different mechanisms of action. SUMMARY: Essential hypertension is a multifactorial and multigenic disorder, which means that various mechanisms contribute to a greater or lesser extent to increasing blood pressure. Drugs combining several mechanisms of action or combinations of antihypertensive drugs with those targeting other risk factors may offer an alternative to reduce overall cardiovascular risk. As we improve our understanding of essential hypertension, it should be possible to develop safer and more effective antihypertensive drugs. The risk/benefit ratio of these new existing antihypertensive drugs will require long-term endpoint assessment studies. PMID- 16046900 TI - New directions in the assessment and treatment of hypertensive heart disease. AB - PURPOSE OF REVIEW: This article will review briefly the emerging new concepts in the diagnosis and treatment of myocardial fibrosis in patients with hypertensive heart disease. RECENT FINDINGS: Although hypertensive heart disease is characterized clinically by development of left-ventricular hypertrophy in the absence of a cause other than arterial hypertension, changes in the composition of myocardial tissue also develop in arterial hypertension leading to structural remodeling of the myocardium (e.g. fibrosis). Recent available data on the determination of serum concentrations of collagen-derived serum peptides and quantitative analysis of echoreflectivity to address the presence of fibrosis in the myocardium of hypertensive patients are promising. In addition, preliminary data suggest that the goal of reducing myocardial fibrosis is achievable in patients with hypertensive heart disease using specific antihypertensive agents (e.g. those interfering with angiotensin II). SUMMARY: The time has come to revisit the current management of hypertensive heart disease simply focused on detecting left-ventricular hypertrophy and controlling blood pressure to reduce left-ventricular mass. It is necessary to develop new approaches aimed at assessing and repairing alterations of myocardial structure and protect myocardial function and, in so doing, to reduce in a more-effective manner adverse risk associated with hypertensive heart disease. PMID- 16046901 TI - Antihypertensive, cardiovascular, and pleiotropic effects of angiotensin-receptor blockers. AB - PURPOSE OF REVIEW: To review the antihypertensive, cardiovascular and pleiotropic effects of angiotensin-receptor blockers (ARBs). RECENT FINDINGS: ARBs are the most recently approved class of antihypertensive agents. They selectively block the angiotensin II type 1 receptor, thus inhibiting most of the deleterious effects of angiotensin II. In addition to blood-pressure control, other benefits may be gained using ARBs. This is because the renin-angiotensin system plays a crucial role in circulatory homoeostasis, and in patients with atherosclerosis, diabetes or hypertension, angiotensin II contributes to the pathophysiology of disease. Evidence-based medicine includes well-controlled studies with mortality and morbidity endpoints in patients with a variety of conditions including hypertension, type 2 diabetes, stroke, renal disease, heart failure, left ventricular hypertrophy and coronary heart diseases. In addition to these hard endpoints, it has been shown that treatment with ARBs prevents the development of type 2 diabetes, ameliorates coronary and peripheral vascular endothelial dysfunction and decreases plasma levels of several markers of vascular inflammation. SUMMARY: ARBs are first-line agents for the treatment of hypertension and cardiovascular diseases. Blocking the renin-angiotensin system with these agents has special advantages due to specific vascular and antiatherosclerotic effects, which contribute to a better cardiovascular and renal protection in patients at risk from or with cardiovascular disease. PMID- 16046902 TI - Cardio-renal insufficiency: the search for management strategies. AB - PURPOSE OF REVIEW: This review focuses on the pathophysiology and treatment of an increasingly common entity, cardio-renal insufficiency. Cardio-renal insufficiency is more than a simultaneous cardiac and renal disease. Patients with this condition live within a fragile equilibrium challenged by the interaction of profibrogenic, atherosclerotic, neurohumoral, and other less known factors. Regarding therapy, the avoidance of oscillations between overfilled decompensated and emptied-overtreated states becomes of critical importance. Particular focus should be paid to personalized treatment, adjusted according to heart and kidney reserve, the predictable complications of therapy, prevention of decompensations, simple measures-based follow-up and alternative procedures. RECENT FINDINGS: Recent studies have established the important repercussions of unbalanced renal function on cardiovascular prognosis. In the heart failure setting, trials involving extensive cohorts of ageing or comorbidity-affected patients are presently under way. Special attention should be paid to recognize the presence of renal failure coexisting with heart failure, especially in patients with deceivingly near-normal plasma creatinine. Formulae to predict creatinine clearance are being increasingly incorporated into daily clinical practice. Disturbed renal function is an underappreciated prognostic factor in heart failure, and renal failure is frequently viewed as a relative contraindication to some proven efficacious therapies. SUMMARY: Cardio-renal insufficiency is an emerging entity, with affected individuals surviving with extreme degrees of simultaneous heart failure and renal failure. Management of the condition is an intellectually demanding process. Crucial to this management is extensive medical expertise and an in-depth understanding of the particular renal, haemodynamic and internal milieu equilibrium of the patients. PMID- 16046903 TI - Antihypertensive treatment: should it be titrated to blood pressure reduction or to target organ damage regression? AB - PURPOSE OF REVIEW: The success of an antihypertensive treatment is difficult to estimate by the extent of blood pressure reduction alone. Above and beyond blood pressure values in an individual patient, it is necessary to monitor the impact of antihypertensive treatment on the development or regression of hypertension induced early end-organ damage. The intermediate objectives or surrogate endpoints may provide additional valuable information about the level of success of a given therapy in a particular patient. Moreover, monitoring intermediate objectives may provide scientific evidence for delineating the best antihypertensive treatment. RECENT FINDINGS: The importance of assessing left ventricular mass, microalbuminuria, carotid wall thickness and the development of new-onset diabetes during antihypertensive treatment has been stressed. Left ventricular hypertrophy during antihypertensive therapy is associated with a lower likelihood of cardiovascular morbidity and mortality, independent of blood pressure lowering and treatment modalities in people with essential hypertension. Likewise, a reduction in urinary albumin excretion seems to be followed by a reduction in cardiovascular morbidity and mortality. Risk implications of reducing carotid wall thickness or of developing new-onset diabetes have been more controversial, although it seems to be that both can have a desirable effect. SUMMARY: The available findings support the necessity to monitor intermediate objectives, aside from blood pressure measurement, during the follow up of hypertensives. PMID- 16046904 TI - Administration-time-dependent effects of antihypertensive treatment on the circadian pattern of blood pressure. AB - PURPOSE OF REVIEW: Many studies show that the extent of the nocturnal blood pressure decline is deterministic of cardiovascular injury and risk. Accordingly, there is growing interest in how to tailor the treatment of hypertensive patients according to their circadian blood-pressure pattern. RECENT FINDINGS: Differences in efficacy depending on the time of day of drug administration lead to differences in effects of antihypertensive drugs on the nocturnal decline relative to the diurnal mean of blood pressure. Thus, bedtime dosing with nifedipine gastrointestinal therapeutic system (GITS) is more effective than morning dosing, while also reducing significantly secondary effects. Bedtime administration of trandolapril results in a safe and effective means of controlling morning blood pressure without inducing excessive reduction nocturnally. The dose-response curve, therapeutic coverage, and efficacy of doxazosin GITS are all markedly dependent on the circadian time of drug administration. Moreover, valsartan administration at bedtime as opposed to upon wakening results in improved day/night blood-pressure ratio, a significant increase in the percentage of controlled patients after treatment, and a significant reduction in urinary albumin excretion. SUMMARY: Nocturnal hypertension increases one's risk of cardiovascular and cerebrovascular events, nephrosclerosis, and progression to end-stage kidney failure in renal patients. Normalization of the circadian blood-pressure pattern is considered an important clinical goal of pharmacotherapy because it may slow the advance of renal injury. Chronotherapy provides a means of individualizing treatment of hypertension according to the circadian blood-pressure profile of each patient, and constitutes a new option in optimizing blood-pressure control and reducing risk. PMID- 16046905 TI - Molecular physiology and the four-component model of renal urate transport. PMID- 16046906 TI - Emerging role of AMP-activated protein kinase in coupling membrane transport to cellular metabolism. AB - PURPOSE OF REVIEW: It has long been recognized that the coupling of membrane transport to underlying cellular metabolic status is critical because transport processes consume a large portion of total cellular energy. Recently, the finely tuned metabolic sensor AMP-activated protein kinase (AMPK) has emerged as a membrane transport regulator, which may permit sensitive transport-metabolism crosstalk. This review will discuss how AMPK may play an important role in the regulation of ion and solute transport across the plasma membrane under both physiological and pathological conditions in epithelia and other tissues. RECENT FINDINGS: Recent studies have found that AMPK, which becomes activated during cellular metabolic stress, promotes the cellular uptake of fuel sources such as glucose and fatty acids to promote ATP generation and inhibits ion-transport proteins such as the cystic fibrosis transmembrane conductance regulator Cl channel and the epithelial Na channel, thereby limiting the dissipation of transmembrane ion gradients. An understanding of the underlying cellular and molecular mechanisms for AMPK-dependent regulation of transport proteins is beginning to emerge. SUMMARY: As earlier studies have focused on the role of nucleotides such as ATP in regulating transport-protein activities, the regulation of membrane transport by AMPK represents a novel and more-sensitive mechanism for the coupling of membrane transport to cellular metabolic status. Identifying new membrane-transport targets of AMPK and elucidating the mechanisms involved in their AMPK-dependent regulation are fruitful areas for new investigation that should yield valuable insights into the pathophysiology of hypoxic and ischemic tissue injury. PMID- 16046907 TI - Integrated physiology of proximal tubular organic anion transport. AB - PURPOSE OF REVIEW: Renal organic anion transport proteins play important roles in the reabsorption and the secretion of endogenous and exogenous compounds. This review focuses on the interpretation of the physiological integration of identified transport molecules in the renal proximal tubules. RECENT FINDINGS: To date, molecular identification of organic anion transport proteins is still continuing: rodent organic anion transporter 5, organic anion-transporting polypeptide 4C1, voltage-driven organic anion transporter 1, multidrug resistance associated protein 4, and sodium-coupled monocarboxylate transporter have yielded additional information in this field. In addition, particularly at the apical membrane of the proximal tubules, the importance of the PDZ (PSD-95, DglA, and ZO 1) binding domain proteins has emerged in the formation of the multimolecular complex as a functional unit of membrane transport. Finally, discovery of dicarboxylate receptors in the renal tubular cells raises the possibility that dicarboxylate anions function as intrarenal signaling molecules. This novel aspect of renal organic anion transport, the potential modulation of signaling via dicarboxylate receptors, may be of significant relevance to renovascular hypertension and other renal diseases. SUMMARY: Comprehensive understanding of the multimolecular complex, which is composed of transporters and their related signaling elements and is supported by the scaffold proteins underneath the plasma membrane, may be useful in clarifying complex transport phenomena such as renal apical organic anion handling. In addition to the recent proteomics approaches and conventional molecular physiology, it is necessary to develop novel methods to analyze the overall function of the multimolecular complex for the post-genomic era. PMID- 16046908 TI - Recent advances in our understanding of intercalated cells. AB - PURPOSE OF REVIEW: This review will summarize newly described novel functions of renal intercalated cells. RECENT FINDINGS: Over the past 20 years, the importance of intercalated cells in the process of renal net acid excretion has been recognized. More recently, many of the molecular mechanisms responsible for this cellular function have been described. Functionally, type A and type B intercalated cells are largely mirror images in that type A intercalated cells are H secreting cells, whereas type B intercalated cells are OH secreting cells. Whether non-A, non-B intercalated cells represent H or OH secreting cells or whether they interconvert between these functions is unclear. Transporters such as pendrin (Slc26a4, Pds), AE1 (Slc4a1), the H-ATPase, and NBC3 (Slc4a7) contribute to the ability of intercalated cells to secrete H or OH equivalents. In addition to mediating secretion of H or OH equivalents, however, intercalated cells also regulate vascular volume, and hence blood pressure, likely by regulating renal Cl excretion. SUMMARY: The molecular mechanisms of net H/OH secretion by intercalated cell subsets have been largely defined over the past decade. Moreover, targeted genetic disruption of these transporters has revealed novel roles, such as blood pressure regulation. Thus, some of these transporters might be the target of future antihypertensive drugs. PMID- 16046910 TI - Molecular pathophysiology of SLC4 bicarbonate transporters. AB - PURPOSE OF REVIEW: Acid-base (H and HCO3) transport in the kidney is crucial for maintaining blood pH, cellular pH and excreting metabolic acid. HCO3 transport in the kidney is mediated by HCO3 transporter proteins which occur in two gene families in humans, vertebrates and invertebrates (SLC4 and SLC26). Since SLC26 transporters have other, non-HCO3 transport functions, this review highlights the history and recent advances in the SLC4 transporters in the kidney. The SLC4 gene and protein family (10 genes) contains three types of HCO3 transporters: Cl-HCO3 exchangers, Na/HCO3 cotransporters and Na-driven Cl-HCO3 exchangers. Function and human chromosomal location have been determined for most members. RECENT FINDINGS: Human mutations in AE1 (SLC4A1) and NBCe1 (SLC4A4) are associated with distal and proximal renal tubular acidosis, respectively. Recent advances include the cellular and biophysical mechanisms by which AE1 and NBCe1 mutations lead to renal disease. Mutational and cellular trafficking studies have begun to elucidate the membrane topology and functional domains of AE1 and NBCe1. Knockout mice for AE2 and NBCn1 do not have obvious renal phenotypes. Recently, SLC4A11 (bicarbonate transporter 1) was shown to function as an electrogenic Na/borate cotransporter unable to transport HCO3 but involved in cell cycle control. SUMMARY: SLC4 HCO3 transporters play critical roles in systemic and cellular pH homeostasis. Most of the SLC4 members are present at some level in the kidney. Future studies will likely continue to make use of knockout animals, for example mice and zebrafish, human mutations or polymorphisms to elucidate the normal and pathophysiologic roles of these proteins. PMID- 16046909 TI - Na+/H+ exchangers: physiology and link to hypertension and organ ischemia. AB - PURPOSE OF REVIEW: Na/H exchangers (NHEs) are ubiquitous proteins with a very wide array of physiological functions, and they are summarized in this paper in view of the most recent advances. Hypertension and organ ischemia are two disease states of paramount importance in which NHEs have been implicated. The involvement of NHEs in the pathophysiology of these disorders is incompletely understood. This paper reviews the principal findings and current hypotheses linking NHE dysfunction to hypertension and ischemia. RECENT FINDINGS: With the advent of large-scale sequencing projects and powerful in-silico analyses, we have come to know what is most likely the entire mammalian NHE gene family. Recent advances have detailed the roles of NHE proteins, exploring new functions such as anchoring, scaffolding and pH regulation of intracellular compartments. Studies of NHEs in disease models, even though not conclusive to date, have contributed new evidence on the interplay of ion transporters and the delicate ion balances that may become disrupted. SUMMARY: This paper provides the interested reader with a concise overview of NHE physiology, and aims to address the implication of NHEs in the pathophysiology of hypertension and organ ischemia in light of the most recent literature. PMID- 16046911 TI - Bibliography. Current world literature. Pharmacology and therapeutics. PMID- 16046912 TI - The evidence base for community-based rehabilitation: a literature review. AB - Literature on community-based rehabilitation (CBR) published between 1978 and 2002 is reviewed to assess the evidence base for CBR. There were 128 articles found that met the criteria set by the authors. The articles have been classified according to the methodology used and the key aspects studied. The review showed an ever-increasing number of publications on CBR. Theory papers and descriptive studies are the most common types of papers in CBR literature. Intervention studies and case reports are relatively rare. No systematic review has yet been carried out although reviews on specific aspects of CBR have become available. The key aspects of 'implementation' and 'stakeholders' are relatively well presented but the numbers of articles on 'participation' and 'use of local resources' are noticeably low. This study reveals that there is no real focus of research in CBR and therefore the evidence base for CBR is fragmented and incoherent on almost all aspects of CBR. It is recommended that comprehensive review studies should be carried out on key aspects of CBR projects. This should be supported by systematic research in CBR projects in order to establish evidence-based practices. PMID- 16046913 TI - Searching for an assessment instrument to determine temporomandibular disorder pain profiles for the purposes of primary health care. AB - The objective was to study the utility of the graded chronic pain (GCP) scale of the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) for the purpose of recognizing TMD pain profiles in primary health care. The GCP scale was used to gather data from 93 consecutive TMD patients seeking treatment in a primary health care setting. The GCP data were compared with pain profiles assessed by scales similar to the more complicated multidimensional pain inventory (MPI). The scales used were able to produce the similar 'dysfunctional profiles' in the TMD patients as the original MPI scale in the sample of chronic pain patients. Also, the original MPI profiles of 'adaptive copers' and 'interpersonally distressed' were produced with distinctive features. The GCP grades distributed almost identically among the subgroups produced by the pain profiles. Most grade I patients were classified as adaptive copers (82--86%), most grade II patients were interpersonally distressed (59%) and all except one of the grade III patients were classified as dysfunctional (83%). The results seem to support the main assumption of our study that the GCP scale is a simple screening tool suitable for identifying primary care TMD patients with various types of pain profiles. PMID- 16046914 TI - Domiciliary therapy during inpatient rehabilitation treatment for patients with an acquired brain injury: a preliminary study. AB - The objective was to assess the feasibility of additional domiciliary treatment for patients with an acquired brain injury while they are still inpatients at a rehabilitation centre. This cohort study included 22 patients with an acquired brain injury (mainly stroke) and with moderate to severe neuropsychological deficits. After two observational home visits for goal setting to determine the aim of the treatment, domiciliary treatment was given once a week in addition to the therapy at the rehabilitation centre. The number of home visits depended on the time interval between admission and time of inclusion in the study. The main outcome measures were (1) a semi-structured interview with patients and partners or children, (2) a questionnaire filled in by the attending therapist(s) before and after each therapeutic home visit and (3) the functional level of the patient. All but one of the patients and all partners and therapists were satisfied with the domiciliary treatment. Most of the main goals were attained during the treatment. The main goals during the domiciliary treatment were in the domains of domestic activities and leisure. A 90--120 min duration of the treatment sessions seems most feasible. It was concluded that domiciliary treatment for inpatients is feasible and seems useful for patients with moderate to severe cognitive impairments. PMID- 16046915 TI - Preliminary validation of the Chinese version of the Pediatric Quality of Life Inventory. AB - The Pediatric Quality of Life Inventory (PedsQL) is a scale for assessing health related quality of life of children and adolescents aged 2--18 years. Three reports of PedsQL for the age ranges 2--4 years and 5--7 years were translated into Chinese and their validities were examined. A total of 186 children and parents were involved in the study. Content validity, test-retest reliability, internal consistency reliability and construct validity were assessed. The correlation of parents' and children's reports was also examined. The results showed that the internal consistency is generally good, test-retest reliability ranged from moderate to good, differences between disabled and non-disabled individuals are significant in total scores and in all subscales except for the physical functioning subscale for the children's self-report for the age ranges of 2-4 years and 5-7 years and the correlation between the reports of the parents and children for the age range of 5-7 years is moderate to high. This suggests that the newly translated Chinese PedsQL for children aged 2-4 years and 5-7 years seems to be reliable and valid to be used as a measure of health-related quality of life in Chinese pediatric research and clinical applications. PMID- 16046916 TI - Health-related quality of life of Turkish stroke survivors. AB - Stroke is a major, chronically disabling neurological disease, which often radically and permanently changes the lives of the victims. Improvement of motor coordination and the degree of independence achieved in activities of daily living have been the usual criteria used to measure outcome in stroke rehabilitation. The objective of our study is to compare health-related quality of life (HRQOL) between stroke patients 6 months after rehabilitation and a control group who did not have any major illness. The associations of several factors with the quality of life were also examined. Sixty consecutive stroke patients and 58 healthy control participants were included in the study. Functional independence measure (FIM), Nottingham health profile (NHP) and demographic variables were used as the main outcome measures. There were significant improvements in both motor coordination and functional status as measured by FIM at discharge and also 6 months after stroke and these variables were significantly associated with the HRQOL. The NHP, which is a validated quality-of-life measurement in Turkish patients, was used and the scores were compared with the healthy control participants. The scores of NHP domains (energy level, emotional reactions, sleep, social isolation and physical mobility) of the stroke patients were higher than those of the healthy group (P<0.01). Age, marital status, lesion side and multiple stroke history showed no significant correlation with NHP scores. Being a woman, lower educational status, tendency to depression and the presence of several comorbidities were significantly correlated with the NHP scores. Stroke survivors in our society have lower HRQOL than healthy individuals. HRQOL is correlated with the functional status in the stroke population. Additionally, the reduced HRQOL after stroke appears to be related to several demographic properties such as sex, education, comorbidities and psychological factors. PMID- 16046917 TI - Development of an exercise testing protocol for patients with a lower limb amputation: results of a pilot study. AB - Due to a decrease in physical activity, lower limb amputees experience a decline in physical fitness. This causes problems in walking with a prosthesis because energy expenditure in walking with a prosthesis is much higher than in walking with two sound legs. Exercise training may therefore increase the functional walking ability of these patients. To generate a safe and effective aerobic training program, exercise testing of amputees is recommended. The objectives of this study were to develop a maximal exercise testing protocol for lower limb amputees and to compare two different testing methods: combined arm-leg ergometry and arm ergometry. The protocols were tested in five amputee patients. Combined ergometry elicited a higher oxygen uptake and heart rate than arm ergometry. Electrocardiography during combined ergometry was easier to read. Combined ergometry was judged most comfortable by the amputees. The exercise testing protocol was useful in lower limb amputees to determine their maximal aerobic capacity and their main exercise limitation. Future exercise training programs may be based on this testing protocol. Combined arm-leg ergometry is appropriate for unilateral amputees without significant claudication of the remaining leg. Continuous arm ergometry is suitable for unilateral amputees with significant claudication of the remaining limb or bilateral amputees. PMID- 16046918 TI - Health-related quality of life of adolescents and young adults 10 years after serious traumatic brain injury. AB - The aim of this study was to investigate health-related quality of life (HRQoL) in a population-based group of young adults with serious traumatic brain injury (TBI) acquired 10 years earlier. In the time period 1987--1991, all 165 residents (<18 years of age) in the south-western health care region of Sweden who had suffered a serious TBI were followed up. Of these, 109 (67%) participated in this follow-up study, which was conducted using the 15-dimension (15D) HRQoL instrument. Their HRQoL was compared with that of 1,039 individuals drawn randomly from the National Population Register for the Finnish Health Care Survey 1995/1996 and matched for age and sex. Nine (mobility, vision, hearing, eating, speech, mental status, depression, distress and usual activities) of the 15 dimensions were significantly aberrant in the TBI group. This TBI group differed markedly from children with organ transplantation, as the transplantation children did not differ from a control group in terms of HRQoL. Compared with other groups of children with congenital or long-lasting conditions, the TBI group had more medical and mental problems. PMID- 16046919 TI - Social inequality in patients' physical and psychological state and participation in rehabilitation after myocardial infarction in Germany. AB - Several Anglo-American studies found that vertical social characteristics made a difference in the utilization of cardiac rehabilitation programmes. A social gradient was also demonstrated for prevalence of risk factors, psychological strain and psychosocial resources. This study investigated social differences for these factors in a group of 536 patients who had suffered from a myocardial infarction. In contrast to findings in other industrialized countries, German patients of higher status groups made less use of cardiac rehabilitative services than patients with a lower and middle class status. However, similar to Anglo American findings, patients with a lower socio-economic status were more likely to be smokers and more likely to be obese than other patients. Also they had higher levels of comorbidity and depression and lower self-efficacy expectations. Thus while there were no social disadvantages in terms of participation in cardiac rehabilitation programmes, which seems to be largely due to the specific characteristics of the German health care system, patients of lower socio economic status seem to have worse baseline conditions and as a result of this a specific need for rehabilitative support. PMID- 16046920 TI - Children's feelings toward parents in the context of parental disability. AB - The purpose of the study was to examine the influence of parents' disability on children's feelings toward their parents. The paper focuses on the comparison of children's feelings toward their parents among school-age children with disabled parents (research group, n=45) and without disabled parents (control group, n=46). A two-dimensional approach was used. This approach is based on the assumption that positive and negative dimensions coexist relatively independently, rather than being polar opposites. The results show that positive and negative feelings toward parents coexist among most of the children in both the groups, but the general intensity of feeling toward parents and the discrepancy between the positive and the negative feeling were higher in the research group than in the control group. The research group was found to express more positive and ambivalent feelings and fewer negative and indifferent feelings than the control group. Moderator effects of age on these feelings in both groups were found as well. The implications of the research findings are discussed and future research is recommended. PMID- 16046921 TI - A preliminary investigation of the efficacy of oral motor exercises for children with mild articulation disorders. AB - Although oral motor therapy is sometimes used to treat articulation disorders in school-age children, several reports question its efficacy. In this case study, four first-grade students, two boys and two girls, received 15 half-hour sessions of oral motor treatment based on Easy Does it for Articulation: An Oral Motor Approach (Strode and Chamberlain,1997). Pre- and post-test measures of the children's articulation indicated no real differences in speech production. These results question the efficacy of general and discrete oral motor exercises because they did not enhance the children's speech production. PMID- 16046922 TI - The effectiveness of picture-fading techniques in a multimedia learning system for teaching Chinese word recognition to students with multiple disabilities. AB - The purpose of the study was to examine the effectiveness of picture-fading techniques in a multimedia learning system for Chinese word-recognition instruction. A single-subject, multiple-probe baseline design across participants was used. Three first-grade students with multiple disabilities participated in this study. The three participants received word-recognition instruction with picture-fading techniques to identify four Chinese printed words. The results of this study demonstrated that all participants could identify the four printed words correctly without the presence of known pictorial prompts. In conclusion, the use of picture-fading techniques in word-recognition instruction can function as a bridge between pictorial prompts and printed words. PMID- 16046923 TI - The combined effects of controlled breathing techniques and ventilatory and upper extremity muscle exercise on cardiopulmonary responses in patients with spinal cord injury. AB - This study investigated the effects of controlled breathing techniques and ventilatory and upper extremity muscle exercise on cardiopulmonary and metabolic functions and exercise tolerance in patients with spinal cord injury (SCI). The design of the study was prospective and was a before-after trial. Twenty patients with SCI were included in the study. Resting pulmonary functions were assessed spirometrically. Cardiopulmonary and metabolic responses to maximum exercise were determined with an electronically braked arm crank ergometer. The expired gases during this exercise were collected and analysed using a computerized gas analysis system. Patients with SCI participated in a 6-week ventilatory and upper extremity muscle exercise program for 1 h, three times per week. At the end of the exercise program, we observed significant improvement in spirometric values, peak oxygen uptake, time to fatigue, peak power output and minute ventilation compared with pre-training values for the patients. PMID- 16046924 TI - Evaluation of skilled arm movements in patients with stroke using a computerized motor-skill analyser for the arm. AB - To study the utility of a new computerized motor-skill analyser (CMSA) for evaluating visuo-motor skilled movements of the arm, we examined (1) the reproducibility of measurement with the CMSA for the arm in 13 healthy participants and 14 patients with stroke and (2) the correlation between motor skills of the affected arm evaluated with the CMSA and clinical tests for upper extremity function in 20 patients with stroke. The CMSA for the arm was used to calculate the accuracy in tracking with the hand. The inter-class correlation coefficients for lap time, geometric trajectory error, trajectory error integrated over time and the marker trajectory error integrated over time were 0.45 (NS)-0.91 (P<0.01) in healthy participants and 0.62 (P<0.05)-0.90 (P<0.01) in patients with stroke. The trajectory error integrated over time and the geometric errors of the affected arm in stroke patients were negatively correlated with the Brunnstrom stage and the simple test for evaluating hand function. The marker trajectory error integrated over time in stroke patients was negatively correlated with the simple test for evaluating hand function. These results suggest that the CMSA for the arm is useful for quantitatively evaluating delicate visuo-motor skills of the arm. PMID- 16046925 TI - Cardiorespiratory fitness of post-stroke patients: as inpatients and as outpatients. AB - The objective of this study was to determine the relationship between disuse weakness due to acute or subacute stroke and cardiorespiratory fitness. The participants were 15 men (average age 48.4 years) who were acute admissions to a hospital specializing in the management of stroke, 11 with cerebral infarction and four with intracerebral hemorrhage. All participants received physiotherapy (40 min/day, 5 days/week) at their bedside until leaving hospital. None of them received physiotherapy in an outpatient clinic. The anaerobic threshold (AT) was measured twice: during admission (an average of 37.3 days after onset) and after discharge (in an outpatient clinic, an average of 37 days after discharge and 42.7 days after the first test). The average AT was 12.5 ml/kg/min in the first test and 14.2 ml/kg/min in the second test (P<0.05). Because ATs in patients were lower after discharge than for healthy control individuals, physiotherapy during admission for stroke should be qualitatively and quantitatively reexamined for the prevention of disuse weakness. After discharge, it is important to give patients a more active rehabilitation program to improve their fitness. PMID- 16046926 TI - Reliability of the Chinese version of the perceived family burden scale. AB - The study examined reliability of the 24-item Chinese version of the perceived family burden scale (CPFBS), which is a measure of behaviors associated with Chinese people with schizophrenia and the impacts of these behaviors on caregivers and relatives. Many patients with mental illness are returning to the community and living with families. Unfortunately, family members who have to live with the patients experience a sense of burden. The CPFBS is able to measure the perceived burden of the family, which enables therapists to formulate the most appropriate family intervention for them. The CPFBS was translated and culturally adapted from the English version. Some 21 main caregivers of individuals with schizophrenia were asked to rate the items on the scale by a competent administrator through a telephone interview (this was carried out twice, with 1 day between each interview). The results showed that CPFBS had high internal consistency (alpha=0.85) and acceptable test-retest reliability (r=0.86). We suggest that the CPFBS is ready to be used by clinicians and researchers in the study on family burden of individuals with schizophrenia in Hong Kong and in Chinese communities in other countries. PMID- 16046928 TI - The stabilometric evaluation of the erect stance. Energy-based optimal control in a numerical model. AB - AIM: This paper proposes an anthropomorphic model developed in a virtual context, useful to simulate the maintenance of the erect stance. Such a model can be built through a suitable software which can identify the geometry of the articular parts and place mechanical constraints (e.g. joints, spherical links, etc.) between the single elements. METHODS: Our approach can be described as the assembly of rigid parts linked at the joints. The control is given by internal torques at the joints and driven by strategy for the balance aimed to the least amount of total muscular energy spent. The anthropomorphic model proposed underlines and solves the typical problems of a system characterized by a multiple number of degree of freedom placed in erect stance and perturbed by phenomena of endogenous and exogenous nature. This model is developed via the coupling of 2 separate bi-dimensional models, one representing the sagittal plane and the other the frontal plane. The numerical tests were validated through experimental tests in a group of healthy volunteers. For this purpose we employed a stabilometric platform (force-plate) to record the statokinesigram (SKG) on the rest surface. RESULTS: The shapes of the stabilometric plots show a good spatial similarity between the experimental and simulated SKG. Both SKGs present a good range in the frontal plane with concentration of points in some areas (or attractors) in the experimental SKG, but not in the simulated one. The tests also showed a postural oscillation at low frequency (= 0.02Hz), probably due to the differential tiredness of groups of muscles or because of a delayed action of the neurological control. CONCLUSIONS: On the numerical simulations, we claim the suitability of the antropomorphic model for a general description of the maintenance of the erect stance. Despite the simplification, with our approach it is possible to simulate some of the main characteristics of the postural act. In particular, the length and the areas of the simulated SKG's are comparable with those of the experimental tests. PMID- 16046929 TI - Rehabilitation after primary total hip replacement. Comparison between Italian and international protocols. AB - AIM: In intensive rehabilitation departments (Code 56) there is a high case-load of patients with recent total hip replacement (THR). Whereas there has been a progressive standardisation and perfecting of prosthetic materials and surgical techniques, time-frames and modalities of rehabilitation programmes are still very variable. Following the Ministerial Guidelines, issued in 1998 by the Italian National Health System, and the introduction of Accreditation Requirements, methods must become more uniform and there must be increased scientific rigour in treatment so as to reduce variability and the subjective nature of the service provided. Ana-lysis of the working methods of large Rehabilitation Centres may help to focus on the problems more clearly and stimulate any improvements that may be required. The study analyses and compares rehabilitation protocols for hip replacement patients adopted in Italian and international rehabilitation centres. METHODS: Thirty-four post-THR rehabilitation protocols were analysed; 14 Italian plus 20 international. RESULTS: The analysis revealed that some factors are unanimously considered important and are therefore codified: 1) posture and positioning; 2) prevention of deep vein thrombosis; 3) rapid return to mobility; 4) education of patients to joint care. CONCLUSIONS: Weight-bearing on the operated limb is not yet a standardised aspect and thus the physiatrist's attention should be focused on this for a return to walking that is safe (for patients and for implanted prosthesis) correct (from the biomechanical and kinematic standpoints) and that plays a normal role in performing activities of daily living. PMID- 16046930 TI - Manual autotraction: preliminary study on the effectiveness of a new device for back pain treatment. AB - AIM: The aim of this study is to assess the efficacy of a new, simple and cheap device of manual autotraction (MAT), in comparison with Natchev's autotraction system. METHODS: Fifty-four patients, with lumbalgia or sciatalgia for more than 4 months and disc herniation or protrusion at computed tomography (CT) or nuclear magnetic resonance (NRM), have been selected, at random, in a randomized control trial. Patients have been treated by MAT or by Natchev's autotraction. Treatments' efficacy has been assessed through the differences in visual analogic scale (VAS) for pain, and Backill scale for disability, before and after therapy (diffVAS and diffBi). The results of the 2 autotraction's devices have been compared in order to distinguish their differences. The results have been considered significant if p<0.05. RESULTS: Both treatments proved to be effective (MAT: diffVAS p<0.0001, diffBackill p<0.001; Natchev's group: diffVAS p<0.0001, diffBackill p<0.001). No significant difference of efficacy emerges between the treatments in diffVAS (p=0.199) and diffBi (p=0.906), and a greater efficacy of both in case of pain with nocturnal aggravation (MAT group: diffVAS: p=0.001; Natchev's group: p=0.001). CONCLUSIONS: The results of this study show an equal efficacy of MAT compared to the known Natchev's system, so it could be equally proposed like a simple conservative treatment in disc protrusion or herniation without surgery indication. PMID- 16046931 TI - Use of botulinum toxin type A in management of adult spasticity. A European consensus statement. PMID- 16046932 TI - Phenomenology of muscle overactivity in the upper motor neuron syndrome. PMID- 16046933 TI - Laboratory analysis and dynamic polyEMG for assessment and treatment of gait and upper limb dysfunction in upper motoneuron syndrome. AB - Multiple muscle agonists and antagonists acting for all the joint movements in the upper and lower limb exist. This redundancy of motor control is very valuable in normal physiology, but when a central nervous system injury with resulting upper motor neuron syndrome takes place, the source of the functional impairment may be difficult to localize. In this paper we discuss the use of gait and motor control analysis studies as a tool particularly useful in determining the specific muscles that may be producing limb dysfunction. We present the most frequent patterns of upper motor neuron dysfunction that affect the upper and lower limb as a result of upper motor neuron syndrome. A case description of the features, the electromyographic patterns, and their functional implications are used. Our objective is to clarify the understanding of these patterns of dysfunction and their focal cause with the intent to improved care of the patient with upper motor neuron syndrome. PMID- 16046934 TI - Selective neuromuscular blocks and chemoneurolysis in the localized treatment of spasticity. AB - Selective neuromuscular blocks and chemoneurolysis are currently the most widespread therapies for treating localized or locoregional spasticity. Both procedures present advantages and disadvantages. The main advantages of BTX-A are its relative ease of use, low incidence of side effects, reversibility and elevated efficacy. Its disadvantages are the limited maximum dose, which does not permit the treatment of many muscles simultaneously, especially if they are large, and its relatively high cost. Phenol neurolysis has a low cost, elevated efficacy in the control of pathologic muscle overactivity, and long duration of effect. Its disadvantages are the risk of injury to the vascular and sensory structures and the difficulty in performing the procedure. The risks associated with neurolysis have led to an increasing interest in and use of BTX-A, making it one of the most widely used therapies in treating localized spasticity. From the perspective of a balanced benefit-risk analysis, a viable option for some cases may be to combine phenol neurolysis for treating spasticity in large proximal muscles and BTX-A for treating hypertonia in small distal muscles. PMID- 16046935 TI - The role of intrathecal baclofen therapy in the upper motor neuron syndrome. AB - This review article will discuss the application of intrathecal baclofen (ITB) therapy in the upper motor neuron syndrome (UMNS). While the UMNS consists of a variety of signs and symptoms, spasticity appears to be the most widely discussed in research and clinical practice. Thus, while a variety of motor disorders result from spasticity and the other components of the UMNS, such as dystonia, rigidity, and co-contraction of agonists and antagonists, we will refer to spasticity as the representative pathology of the UMNS. The term spasticity will be used in this discussion as if it were synonymous to the UMNS, because it is the term used in most published research and papers. This does not necessarily mean that the other features of the UMNS are less important. Publications in the use of ITB in the pediatric population, especially cerebral palsy, abound, but this paper will focus on UMNS in adults. In addition to reviewing the process of ITB management, from patient selection to rehabilitation, topics of practical interest to clinicians will be discussed. PMID- 16046936 TI - Neuro-orthopedic management of the dysfunctional extremity in upper motor neuron syndromes. AB - Orthopedic surgery can help restore extremity function to many patients with upper motor neuron syndromes (UMN). Impairments are divided into those that cause problems with the active function of the extremity versus those that impede passive function. Limb deformities commonly result from both dynamic (spastic) and static (contractural) components. Clinical examination supplemented with dynamic electromyographic studies provides the optimal information for planning the most effective surgical procedures. In the upper extremity, selective lengthening of the shoulder adductors and extensors combined with elbow flexor lengthening can improve forward reach. Lengthening of the forearm pronators and finger flexors will improve hand use. In the lower extremity, standing balance is improved with widening the base of support by correcting hip adduction contractures and equinovarus foot deformities. Improvement of knee flexion during swing phase by a rectus femoris to gracilis transfer will enhance the fluidity and efficiency of walking. Correction of hip and knee flexion contractures will allow a upright posture and dramatically decrease the energy requirement of walking. PMID- 16046937 TI - Availability of emergency contraception: a survey of hospital emergency department staff. AB - STUDY OBJECTIVE: I investigate accessibility of emergency contraception pills at hospital emergency departments and survey staff at Catholic and non-Catholic hospitals across the United States. More specifically, I sought to report the likelihood that a woman calling a hospital and seeking emergency contraception could access the medication; (2) if emergency contraception is not provided, whether hospital staff would provide a referral to another facility; and (3) the outcome of the referral process. METHODS: Using a "mystery client" approach, I telephoned staff at all 597 Catholic hospitals in the United States and at 17% of non-Catholic hospitals (n=615). I used this interviewing method to reflect the experience of a laywoman calling to inquire about the availability of emergency contraception. RESULTS: I found that staff at 42% of non-Catholic hospitals and 55% of Catholic hospitals said that they do not dispense emergency contraception, even in cases of sexual assault. Overall, more respondents at Catholic hospitals (23%) reported that they provide emergency contraception only to victims of sexual assault compared with staff at non-Catholic hospitals (17%). Among staff who said that their hospital does not provide emergency contraception under any circumstances, only about half gave callers a valid referral, and most referrals were ineffective. CONCLUSION: To improve women's access to emergency contraception, hospitals can (1) use collaborative drug-therapy agreements to enable hospital pharmacies to dispense emergency contraception without a prescription, (2) develop and communicate written policies that support provision of emergency contraception, and (3) encourage health care providers who observe religious or ethical guidelines to provide effective referrals for women seeking emergency contraception. PMID- 16046938 TI - Emergency contraception: what should our patients expect? PMID- 16046939 TI - Images in emergency medicine. Lemierre syndrome. PMID- 16046940 TI - The impact of aeromedical response to patients with moderate to severe traumatic brain injury. AB - STUDY OBJECTIVE: Aeromedical crews offer an advanced level of practice and rapid transport to definitive care; however, their efficacy remains unproven. Previous studies have used relatively small sample sizes or have been unable to adequately control for the effect of other potentially influential variables. Here we explore the impact of aeromedical response in patients with moderate to severe traumatic brain injury. METHODS: This was a retrospective analysis using our county trauma registry. All patients with head Abbreviated Injury Score of 3 or greater were included; interfacility transfers were excluded. The impact of aeromedical response was determined using logistic regression, adjusting for age, sex, mechanism, preadmission Glasgow Coma Scale score, head Abbreviated Injury Score, Injury Severity Score, and the presence of preadmission hypotension. Propensity scores were used to account for variability in selection of patients to undergo air versus ground transport. Patients with moderate and severe traumatic brain injury, as defined by head Abbreviated Injury Score and Glasgow Coma Scale score, were compared. Finally, aeromedical patients undergoing field intubation were compared with ground patients undergoing emergency department (ED) intubation. RESULTS: A total of 10,314 patients meeting all inclusion and exclusion criteria and with complete data sets were identified and included 3,017 transported by aeromedical crews. Overall mortality was 25% in the air- and ground-transported cohorts, but outcomes were significantly better for the aeromedical patients when adjusted for age, sex, mechanism of injury, hypotension, Glasgow Coma Scale score, head Abbreviated Injury Score, and Injury Severity Score (adjusted odds ratio [OR] 1.90; 95% confidence interval [CI] 1.60 to 2.25; P<.0001). Good outcomes (discharge to home, jail, psychiatric facility, rehabilitation, or leaving against medical advice) were also higher in aeromedical patients (adjusted OR 1.36; 95% CI 1.18 to 1.58; P<.0001). The primary benefit appeared to be in more severely injured patients, as reflected by head Abbreviated Injury Score and Glasgow Coma Scale score. Improved survival was also observed for air-transported patients intubated in the field versus ground transported patients given emergency intubation in the ED (adjusted OR 1.42; 95% CI 1.13 to 1.78; P<.001). CONCLUSION: Here we analyze a large database of patients with moderate to severe traumatic brain injury. Aeromedical response appears to result in improved outcomes after adjustment for multiple influential factors in patients with moderate to severe traumatic brain injury. In addition, out-of-hospital intubation among air-transported patients resulted in better outcomes than ED intubation among ground-transported patients. Patients with more severe injuries appeared to derive the greatest benefit from aeromedical transport. PMID- 16046941 TI - Prospective performance assessment of an out-of-hospital protocol for selective spine immobilization using clinical spine clearance criteria. AB - STUDY OBJECTIVE: We determine whether the use of an emergency medical services (EMS) protocol for selective spine immobilization would result in appropriate immobilization without spinal cord injury associated with nonimmobilization. METHODS: A 4-year prospective study examined EMS and hospital records for patients after the implementation of an EMS protocol for selective spine immobilization. EMS personnel were trained to perform and document a spine injury assessment for out-of-hospital trauma patients with a mechanism of injury judged sufficient to cause a spine injury. The assessment included these clinical criteria: altered mental status, evidence of intoxication, neurologic deficit, suspected extremity fracture, and spine pain or tenderness. The protocol required immobilization for patients with a positive assessment on any of those criteria. Outcome characteristics included the presence or absence of spine injury and spine injury management. RESULTS: The study collected data on 13,483 patients; 126 of the patients were subsequently excluded from the study because of incomplete data, leaving a study sample of 13,357 patients with complete data. Spine injuries were confirmed in the hospital records for 3% (n=415) of patients, including 50 patients with cord injuries and 128 patients with cervical injuries. Sensitivity of the EMS protocol was 92% (95% confidence interval [CI] 89.4 to 94.6%) resulting in nonimmobilization of 8% of the patients with spine injuries (33 of 415). None of the nonimmobilized patients sustained cord injuries. The specificity was 40% (95% CI 38.9 to 40.5%). CONCLUSION: The use of our selective immobilization protocol resulted in spine immobilization for most patients with spine injury without causing harm in cases in which spine immobilization was withheld. PMID- 16046942 TI - Automated external defibrillators: to what extent does the algorithm delay CPR? AB - STUDY OBJECTIVE: Maximizing cardiopulmonary resuscitation (CPR) during resuscitation may improve survival. Resuscitation protocols stack up to 3 shocks to achieve defibrillation, followed by an immediate postdefibrillation pulse check. The purpose of this study is to evaluate outcomes of rhythm reanalyses immediately after shock, stacked shocks, and initial postshock pulse checks in relation to achieving a pulse and initiating CPR. METHODS: We conducted an observational study of patients with ventricular fibrillation treated by first tier emergency medical services (EMS). We collected data from EMS, dispatch, and hospital records. Additionally, we analyzed automatic external defibrillator recordings to determine the proportion of cardiac arrest victims who were defibrillated and achieved a pulse according to shock number (single versus stacked shock), proportion of victims with a pulse during the initial postdefibrillation pulse check, and interval from initial shock to CPR. RESULTS: The study included 481 cardiac arrest subjects. Automatic external defibrillators terminated ventricular fibrillation with the initial shock in 83.6% (n=402) of cases. A second shock terminated ventricular fibrillation in an additional 7.5% (n=36) of cases, and a third shock terminated ventricular fibrillation in 4.8% (n=23) of cases. The initial sequence of 3 shocks failed to terminate ventricular fibrillation in 4.1% (n=20) of cases. In total, automatic external defibrillators performed 560 rhythm reanalyses during the initial shock sequence and delivered 122 "stacked" shocks. Termination of ventricular fibrillation was not synonymous with return of a pulse. The initial shock produced a pulse that was eventually detected in 21.8% (105/481) of cases. Stacked shocks produced a pulse in 10.7% (13/122) of cases. For the 24.5 % (n=118) of cases in which a pulse returned, the pulse was detected during the initial postshock pulse check only 12 times, or 2.5% of all cases. The median interval from initial shock until CPR was 29 (23,41) seconds. CONCLUSION: Rhythm reanalyses, stacked shocks, and postshock pulse checks had low yield for achieving or detecting return of a pulse. CPR was not initiated until 29 seconds after the initial shock. PMID- 16046943 TI - Workplace violence: a survey of emergency physicians in the state of Michigan. AB - STUDY OBJECTIVE: We seek to determine the amount and type of work-related violence experienced by Michigan attending emergency physicians. METHODS: A mail survey of self-reported work-related violence exposure during the preceding 12 months was sent to randomly selected emergency physician members of the Michigan College of Emergency Physicians. Work-related violence was defined as verbal, physical, confrontation outside of the emergency department (ED), or stalking. RESULTS: Of 250 surveys sent, 177 (70.8%) were returned. Six were blank (3 were from retired emergency physicians), leaving 171 (68.4%) for analysis. Verbal threats were the most common form of work-related violence, with 74.9% (95% confidence interval [CI] 68.4% to 81.4%) of emergency physicians indicating at least 1 verbal threat in the previous 12 months. Of the emergency physicians responding, 28.1% (95% CI 21.3% to 34.8%) indicated that they were victims of a physical assault, 11.7% (95% CI 6.9% to 16.5%) indicated that they were confronted outside of the ED, and 3.5% (95% CI 0.8% to 6.3%) experienced a stalking event. Emergency physicians who were verbally threatened tended to be less experienced (11.1 versus 15.1 years in practice; mean difference -4.0 years [95% CI -6.4 to -1.6 years]), as were those who were physically assaulted (9.5 versus 13.1 years; mean difference -3.6 years [95% CI -5.9 to -1.3 years]). Urban hospital location, emergency medicine board certification, or on-site emergency medicine residency program were not significantly associated with any type of work-related violence. Female emergency physicians were more likely to have experienced physical violence (95% CI 1.4 to 5.8) but not other types of violence. Most (81.9%; 95% CI 76.1% to 87.6%) emergency physicians were occasionally fearful of workplace violence, whereas 9.4% (95% CI 5.0% to 13.7%) were frequently fearful. Forty-two percent of emergency physicians sought various forms of protection as a result of the direct or perceived violence, including obtaining a gun (18%), knife (20%), concealed weapon license (13%), mace (7%), club (4%), or a security escort (31%). CONCLUSION: Work-related violence exposure is not uncommon in EDs. Many emergency physicians are concerned about the violence and are taking measures, including personal protection, in response to the fear. PMID- 16046944 TI - Drug and alcohol use in emergency medicine residency: an impaired resident's perspective. AB - We share the personal experience of an impaired resident who successfully completed rehabilitation and is about to graduate from an emergency medicine program and perform a brief literature review on drug and alcohol abuse in emergency medicine residencies. Residents in general are less likely than their same-age peers to abuse drugs, but a significant minority starts using drugs during residency. Emergency medicine residents have higher rates of substance use than residents in other specialties and are more likely to report current use of cocaine and marijuana. PMID- 16046945 TI - Road traffic injuries in Lazio, Italy: a descriptive analysis from an emergency department-based surveillance system. AB - STUDY OBJECTIVE: Road crashes are the leading cause of death for young adults in industrialized countries. In most countries, the only data available are the police-based statistics, which report the number of deaths and injuries, without any information about severity or diagnosis. We describe the results of an emergency department (ED)--based integrated surveillance system of road-crash health consequences in the Lazio region (5.3 million inhabitants) for 2000. METHODS: We identified all ED visits in the emergency database and then linked them with hospital discharges and mortality registry. Deaths that occurred at the site of the crash were identified in the mortality registry. Cases are classified according to body region affected, nature of the injury, and severity. RESULTS: We found 146,600 cases, for an overall incidence of 2,700 per 100,000 and a peak of 8,000 per 100,000 in 20- to 24-year-old men. There were 597 fatalities for men and 205 female fatalities, with an overall mortality of 15.9 per 100.000 and a hospitalization rate of 224 per 100,000. CONCLUSION: Our surveillance system reported a 6-times-higher incidence of road-traffic injuries, with 139 more fatalities than the national statistics of the same year. The underreporting in police reports and the absence of diagnostic information reveal the need for health-based surveillance systems of road-traffic injuries. PMID- 16046946 TI - Counting road traffic deaths and injuries: poor data should not detract from doing something! PMID- 16046947 TI - A positive blood alcohol concentration is the main predictor of recurrent motor vehicle crash. AB - STUDY OBJECTIVE: The identification of risk factors for recurrent motor vehicle crashes is the basis for prevention, but few studies have been published on predictors of recurrence. Our objective is to determine the main variables predicting recurrent crashes in subjects attending an emergency department (ED) for injuries after motor vehicle crash. METHODS: During a 5-year follow-up period, we studied 2,354 consecutive adult subjects treated in the ED after a motor vehicle crash in 1998. The variables of the original event were tested for predicting recurrence in a Cox proportional hazard model. RESULTS: During follow up, 390 of 2,325 (16.8%) survivors were treated for injury after a new crash. The overall event rate was 34 per 1,000 subject-years. Four variables (age < or =32 years, male sex, nighttime crash, and blood alcohol concentration >50 mg/dL) were identified as independent predictors of recurrent crash. After adjustment for sex, age, and nighttime, alcohol was the leading predictor (relative risk 3.73; 95% confidence interval 3.00 to 4.64). In the presence of the 4 variables, the recurrence rate was as high as 145 (117 to 175) events per 1,000 subject-years, and alcohol per se accounted for more than 75% of events. In the absence of the 4 variables, the rate was as low as 11 (7 to 17) events per 1,000 subject-years. CONCLUSION: Alcohol was the most powerful behavioral factor predicting recurrent events in subjects treated in an ED for injury after motor vehicle crash, along with young age, male sex, and nighttime. PMID- 16046948 TI - National Highway Traffic Safety Administration (NHTSA) notes. Knee-thigh-hip injuries in frontal crashes. PMID- 16046950 TI - Can wrist blood pressure oscillometer be used for triage in an adult emergency department? AB - STUDY OBJECTIVE: We compare the performance of a wrist blood pressure oscillometer with the mercury standard in the triage process of an emergency department (ED) and evaluate the impact of wrist blood pressure measurement on triage decision. METHODS: Blood pressure was successively measured with the standard mercury sphygmomanometer and with the OMRON-RX-I wrist oscillometer in a convenience sample of 2,493 adult patients presenting to the ED with non-life threatening emergencies. Wrist and mercury measures were compared using criteria of the Association for the Advancement of Medical Instrumentation (AAMI) and the British Hypertension Society (BHS). The impact on triage decisions was evaluated by estimating the rate of changes in triage decisions attributable to blood pressure results obtained with the wrist device. RESULTS: Wrist oscillometer failed to meet the minimal requirements for recommendation by underestimating diastolic and systolic blood pressure. Mean (+/-SD) differences between mercury and wrist devices were 8.0 mm Hg (+/-14.7) for systolic and 4.2 mm Hg (+/-12.0) for diastolic measures. The cumulative percentage of blood pressure readings within 5, 10, and 15 mm Hg of the mercury standard was 32%, 58%, and 72% for systolic, and 40%, 67%, and 83% for diastolic measures, respectively. Using the wrist device would have erroneously influenced the triage decision in 7.6% of the situations. The acuity level would have been overestimated in 2.2% and underestimated in 5.4% of the triage situations. CONCLUSION: The performance of the OMRON-RX-I wrist oscillometer does not fulfill the minimum criteria of AAMI and BHS compared with mercury standard in the ED triage setting. PMID- 16046951 TI - Emergencies in space. AB - Manned spaceflight is inherently risky and results in unique problems from a trauma and medical perspective. Emergency care under these special physiologic and environmental conditions calls for novel techniques for diagnosis and therapy. PMID- 16046952 TI - Practical implementation of the Guidelines for Unstable Angina/Non-ST-Segment Elevation Myocardial Infarction in the emergency department. AB - In the United States each year, >5.3 million patients present to emergency departments with chest discomfort and related symptoms. Ultimately, >1.4 million individuals are hospitalized for unstable angina and non-ST-segment elevation myocardial infarction. For emergency physicians and cardiologists alike, these patients represent an enormous challenge to accurately diagnose and appropriately treat. This update of the 2002 American College of Cardiology/American Heart Association Guidelines for the Management of Patients with Unstable Angina and Non-ST-Segment Elevation Myocardial Infarction (UA/NSTEMI) provides an evidence based approach to the diagnosis and treatment of these patients in the emergency department, in-hospital, and after hospital discharge. Despite publication of the guidelines several years ago, many patients with UA/NSTEMI still do not receive guidelines-indicated therapy. PMID- 16046953 TI - Are we putting the cart ahead of the horse: who determines the standard of care for the management of patients in the emergency department? PMID- 16046954 TI - Defining the outcome. PMID- 16046955 TI - Images in emergency medicine. Group B streptococcus. PMID- 16046956 TI - Paper case scenarios in the assessment of triage tools. PMID- 16046957 TI - Triage questions. PMID- 16046958 TI - Emergency department triage: why we need a research agenda. PMID- 16046959 TI - Loxoscelism and necrotic arachnidism: more myths and minor corrections. PMID- 16046960 TI - Intravenous administration of N-acetylcysteine. PMID- 16046961 TI - A Chinese herbal weight loss product adulterated with fenfluramine. PMID- 16046962 TI - Fundamentals of extracoronal tooth preparation. Part I. Retention and resistance form. 1974. PMID- 16046963 TI - Management of obstructive sleep apnea in an edentulous patient with a mandibular advancement splint: a clinical report. AB - Literature on the treatment of obstructive sleep apnea in edentulous patients with a mandibular advancement splint is sparse. This clinical report describes a clinical and laboratory method of splint fabrication and discusses the rationale for its use. PMID- 16046964 TI - A clinical evaluation of fixed partial denture impressions. AB - STATEMENT OF PROBLEM: Providing the dental laboratory with an accurate replication of the hard and soft tissue of a patient is important. Therefore, it is essential to examine whether clinicians critically evaluate impressions routinely before sending them to the laboratory. PURPOSE: This study evaluated the quality of impressions sent to commercial laboratories for the fabrication of fixed partial dentures (FPD) by describing the frequency of clinically detectable errors and by analyzing correlations between the various factors involved. MATERIAL AND METHODS: A total of 193 FPD impressions were evaluated, immediately after arrival at 11 dental laboratories, by 3 calibrated examiners. The impression technique and material used, tray type, and number of prepared units were recorded for each impression. Data relating to errors and faults, including defects in material polymerization, retention to tray, tissue contact by tray, crucial areas beyond tray borders, heavy-bodied material exposure through the wash material (for double-step impressions), inadequate union of materials, retraction cords embedded in impressions, and air bubbles, voids, or tears along the margin were also documented. The data were analyzed with the Pearson chi square test (alpha = .05). RESULTS: Of the impressions, 89.1% had 1 or more observable errors. Significant correlations were found between material type and voids or tears at the finish line (Rv = 0.17, P < .025) and between material type and polymerization problems (Rv = 0.223, P < .004). CONCLUSIONS: Within the limitations of this study, impressions made with polyethers had the most detectable errors, followed by condensation-type silicones. The high frequency of detectable errors found in impressions sent for FPD fabrication is of concern. PMID- 16046965 TI - Effects of different drinks on stainability of resin composite provisional restorative materials. AB - STATEMENT OF PROBLEM: The ability of tooth-colored restorative materials for provisional restorations to resist stains is important when interim prostheses are worn for a long period. Discoloration of provisional restorative materials may result in patient dissatisfaction and additional time and expense for replacement. However, the effect of different staining agents on the color difference of resin composite restorative materials has not been completely clarified. PURPOSE: The purpose of this study was to evaluate the stainability of auto- and light-polymerized composite provisional restorative materials, reinforced microfill, and microhybrid resin composite restorative materials upon exposure to different staining agents. MATERIAL AND METHODS: Forty-five cylindrical specimens (15 x 2 mm) were prepared for each of an autopolymerized bis-acryl composite provisional restorative material (Protemp II), a light polymerized composite provisional restorative material (Revotek LC), a reinforced microfill (Micronew), and a microhybrid composite (Filtek Z250, Herculite XRV) restorative material, using a brass mold. The specimens were wet-ground with 1000 grit silicon carbide abrasive paper for 10 seconds. The 5 restorative material specimens were divided into 9 groups (n = 5) and stored for 24 hours at 37 degrees C in different types of solutions: water, coffee, coffee with sugar, tea, tea with sugar, coffee with artificial creamer and sugar, cola, red wine, or sour cherry juice. Color of all specimens was measured before and after exposure with a colorimeter using CIE L * a * b * relative, and color changes (DeltaE * ) were then calculated. The data were analyzed with a 2-way analysis of variance (ANOVA), and mean values were compared by the Tukey HSD test (alpha = .05). RESULTS: The interaction of provisional restorative materials and staining agents was statistically significant ( P = .0001). For the 5 restorative materials tested, the lowest DeltaE * values were observed in the water, cola, and sour cherry juice groups. The highest color difference for all restorative materials was observed in the red wine groups. For microhybrid composite materials and light-polymerized composite provisional material, when tea and coffee groups with and without sugar were compared, both groups with sugar demonstrated a higher color difference than without sugar. When comparing the 5 different restorative materials, the reinforced microfill material group (Micronew) demonstrated significantly less color change than the other materials tested. The highest color difference in this study was observed for specimens in the light polymerized composite provisional material group (Revotek LC). CONCLUSION: The reinforced microfill restorative material tested was found significantly more color stable than the autopolymerized bis-acryl, light-polymerized composite provisional restorative materials, and microhybrid composites tested. The largest color difference was observed in the light-polymerized composite provisional material. The presence of sugar in coffee and tea increased the color difference compared to coffee or tea without sugar for light-polymerized composite provisional material and microhybrid composites. PMID- 16046966 TI - Flexural strength of a layered zirconia and porcelain dental all-ceramic system. AB - STATEMENT OF PROBLEM: New processing techniques have facilitated the use of zirconia core materials in all-ceramic dental prostheses. Zirconia has many potential advantages compared to existing core materials; however, its performance when layered with porcelain has not been evaluated. PURPOSE: This study investigated the strength of a wide variety of layered zirconia and porcelain beams to determine whether the inclusion of zirconia cores results in improved strength. MATERIAL AND METHODS: Eight types of layered or simple zirconia and porcelain beams (n = 10), approximately fixed partial denture-size, were made of a tetragonal polycrystalline zirconium dioxide partially stabilized with yttria core (Lava System Frame) and a feldspathic dental porcelain (Lava Ceram veneer ceramic). Elastic moduli of the materials were measured using an acoustic method. Maximum force and modulus of rupture were determined using 3 point flexural testing and a universal testing machine. Descriptive statistical methods were used. RESULTS: Beams with porcelain tensile surfaces recorded mean tensile strengths or moduli of rupture from 77 to 85 MPa, whereas beams with zirconia tensile surfaces recorded moduli of rupture almost an order of magnitude higher, 636 to 786 MPa. The elastic moduli of the porcelain and zirconia materials were 71 and 224 GPa, respectively. Crack propagation following initial tensile cracking often involved the porcelain-zirconia interface, as well as bulk porcelain and zirconia. CONCLUSION: The layered zirconia-porcelain system tested recorded substantially higher moduli of rupture than have been previously reported for other layered all-ceramic systems. PMID- 16046967 TI - Longevity and failure load of ceramic veneers with different preparation designs after exposure to masticatory simulation. AB - STATEMENT OF PROBLEM: Laminate veneers are widely used in the management of unesthetic anterior teeth. However, limited information is available regarding the influence of preparation design on longevity of ceramic veneers. PURPOSE: This study evaluated the influence of preparation design on longevity and failure load of ceramic veneers bonded to human maxillary central incisors after cyclic loading and thermal cycling in a dual-axis masticatory simulator. MATERIAL AND METHODS: Sixty-four caries-free maxillary central incisors were divided into 4 groups (n = 16). The control group remained unprepared (NP). For Group WP, a window preparation was made. Specimens in Group IOP were prepared with an incisal overlap of 2 mm without palatal chamfer. For Group CVP, specimens were prepared with a complete-veneer design of 3-mm incisal reduction and 2-mm palatal extension. Forty-eight IPS Empress 1 ceramic veneers were bonded adhesively with dual-polymerizing composite (Variolink II). All specimens were subjected to cyclic mechanical loading (1.2 million cycles, cycle frequency 1.3 Hz, invariable palatal load 49 N) and thermal cycling (5 degrees C-55 degrees C, dwell time 60 seconds, 5500 cycles) in a masticatory simulator. Failure was defined by bulk fracture of a specimen. Subcritical crack patterns were observed. Surviving specimens were loaded in a universal testing machine until fracture. The failure load values (N) (1.5 mm/min crosshead speed) were automatically recorded by controlling software. Statistical analysis of data was performed by Kruskal Wallis analysis of variance (alpha = .05) and pairwise Wilcoxon rank sum tests (alpha = .05). RESULTS: Three specimens from group NP, 1 specimen each from the WP and CVP groups, and 2 specimens from group IOP fractured during fatigue. After 1.2 million cycles, the highest crack rates were observed for complete veneers and originated in the palatal concavity extending to the facial surface. The median (interquartile range = x .25 - x .75 ) failure loads (N) were as follows: NP 713.3 (404.4-777.1), WP 549.5 (477.5-597.7), IOP 695.3 (400.0-804.6), and CVP 519.2 (406.1-732.9). No significant differences in longevity and failure load were demonstrated between natural teeth and teeth restored with ceramic veneers ( P = .555). CONCLUSION: Maxillary teeth restored with the 3 types of IPS Empress 1 veneers showed fracture resistance similar to that of unprepared incisors ( P = .555). PMID- 16046969 TI - Annual review of selected dental literature: report of the Committee on Scientific Investigation of the American Academy of Restorative Dentistry. PMID- 16046968 TI - Effect of high-powered LED polymerization on the shear bond strength of a light polymerized resin luting agent to ceramic and dentin. AB - STATEMENT OF PROBLEM: A newly introduced high-powered light-emitting diode (LED) light-polymerization unit with various polymerization modes is purported to polymerize dental resins more quickly than standard units. However, there is insufficient information about the effects of this type of light source and its polymerization modes on resin luting agents (RLAs). PURPOSE: This in vitro study evaluated the effects of different modes of a high-powered LED polymerization unit on the shear bond strength of a light-polymerized RLA to ceramic and dentin. MATERIAL AND METHODS: Sixty ceramic cylinders (3 x 3 mm) were fabricated from a heat-pressed ceramic (IPS Empress 2). Specimen surfaces were abraded using 600 grit silicon carbide paper and airborne-particle abraded with 50-mum desiccant alumina particles, cleaned ultrasonically, etched using hydrofluoric acid, and treated with a silane coupling agent (Ceramic primer). Sixty noncarious, freshly extracted, permanent human molar teeth were embedded in autopolymerizing acrylic resin, and their dentin surfaces were wet-ground using a grinding and polishing machine fitted with 180-grit abrasive disks. The ground dentin surfaces were then etched with 37% phosphoric acid gel, and an adhesive agent (Single Bond) was applied. Ceramic specimens (n = 15) were bonded to dentin surfaces with a dual initiated RLA (Rely-X ARC) and individually polymerized by 1 of 4 different modes, as follows: Halogen standard mode (Control) (600 mW/cm2 for 40 seconds); high-powered LED fast mode (1100 mW/cm2 for 10 seconds); high-powered LED pulse mode (1100 mW/cm2 for 10 seconds); and high-powered LED exponential mode (1100 mW/cm2 for 20 seconds). Cemented specimens were subjected to shear loading until fracture using a universal testing machine. A stereomicroscope (x25) was used to identify the mode of fracture. Bond strength (MPa) data were analyzed using 1-way analysis of variance and the Tukey HSD test (alpha = .05). RESULTS: Specimens polymerized using halogen standard mode (23.9 +/- 1.3 MPa) and LED exponential mode (23.0 +/- 1.1 MPa) had significantly higher ( P < .001) mean shear bond strengths compared with both LED fast (15.1 +/- 1.3 MPa) and pulse (14.6 +/- 1.3 MPa) modes. The Tukey HSD tests showed no significant differences in shear bond strength between specimens polymerized using high-powered LED exponential mode and halogen standard mode. Most failures were adhesive failures at the dentin-RLA interface or the RLA-ceramic interface in specimens polymerized using high powered LED fast or pulse modes. CONCLUSION: Within the limitations of this study, the shear bond strength of an RLA to ceramic and dentin was found to be similar when polymerized using halogen light in standard mode and high-powered LED light in exponential mode, whereas shear bond strength was significantly lower when polymerized using LED in fast or pulse mode. PMID- 16046970 TI - Osseointegrated craniofacial implants in the rehabilitation of orbital defects: an update of a retrospective experience in the United States. AB - STATEMENT OF PROBLEM: Since their introduction, craniofacial implants have been used in prosthetic rehabilitation of facial defects. The literature, however, indicates marked variability in outcomes using implants for the retention of orbital prostheses. PURPOSE: A multicenter report updating the experience in the United States with the use of craniofacial implants for prosthetic rehabilitation of orbital defects is presented. MATERIAL AND METHODS: Surveys were sent to clinicians at 25 centers where maxillofacial prosthetic treatment is provided to obtain retrospective data regarding patients who completed implant-retained orbital prosthetic rehabilitation. Data on implant placement location, radiation treatment history, and use of hyperbaric oxygen therapy were collected and assessed in relationship to implant survival over time. The Kaplan-Meier life table and Wilcoxon analyses (alpha = .05) were used to assess the significance of the findings. RESULTS: Ten centers responded, providing data suitable for statistical analysis on 153 implants placed to retain 44 orbital prostheses and followed for a mean period of 52.6 months. Forty-one implant integration failures occurred during this follow-up period, resulting in an overall integration survival rate of 73.2%. No significant relationship was found between radiation treatment history, hyperbaric oxygen therapy history, or implant placement location and implant survival. Individual responses revealed large variability between reporting centers in treatment outcomes. CONCLUSION: Craniofacial implants may offer marked benefits in the prosthetic rehabilitation of orbital defects when compared to conventional adhesive retention designs. However, questions remain regarding long-term predictability and the impact specific factors may have on treatment outcomes. Insufficient data is currently available from which to draw statistically meaningful conclusions. The establishment of a national database designed to acquire adequate data to assess treatment outcomes is recommended. PMID- 16046971 TI - Effect of microwave disinfection on the hardness and adhesion of two resilient liners. AB - STATEMENT OF PROBLEM: Microwave irradiation has been suggested for denture disinfection. However, the effect of this procedure on the hardness and bond strength between resilient liners and denture base acrylic resin is not known. PURPOSE: This study evaluated the effect of water storage time and microwave disinfection on the hardness and peel bond strength of 2 silicone resilient lining materials to a heat-polymerized acrylic resin. MATERIAL AND METHODS: Acrylic resin (Lucitone 199) specimens (75 x 10 x 3 mm) were stored in water at 37 degrees C (2 or 30 days) before bonding (n = 160). The resilient lining materials (GC Reline Extra Soft and Dentusil) were bonded to the denture base and divided into the following 4 groups (n = 10): Tests performed immediately after bonding (control); specimens immersed in water (200 mL) and irradiated twice, with 650 W for 6 minutes; specimens irradiated daily for 7 total cycles of disinfection; specimens immersed in water (37 degrees C) for 7 days. Specimens were submitted to a 180-degree peel test (at a crosshead speed of 10 mm/min) and the failure values (MPa) and mode of failure were recorded. Pretreatment and posttreatment hardness measurements (Shore A) of the resilient materials were also performed. Three-way analysis of variance, followed by the Tukey HSD test, was performed (alpha = .05). RESULTS: The analysis revealed that, for all conditions, the mean failure strengths of GC Reline Extra Soft (0.95-1.19 MPa) were significantly higher ( P < .001) than those of Dentusil (0.45-0.50 MPa). The adhesion of the liners was not adversely affected by water storage time of Lucitone 199 or microwave disinfection. All peel test failures were cohesive. There was a small but significant difference ( P < .001) between the pretreatment (34.33 Shore A) and posttreatment (38.69 Shore A) hardness measurements. CONCLUSION: Microwave disinfection did not compromise the hardness of either resilient liners or their adhesion to the denture base resin Lucitone 199. PMID- 16046972 TI - Compatibility of mechanical and computerized axiographs: a pilot study. AB - STATEMENT OF PROBLEM: There is little known information comparing axiographic data obtained with a mechanical device and data obtained with a computerized device. However, long-term follow-up of patients may necessitate comparison of previously made mechanical axiographic data with records obtained through computer-aided axiography. PURPOSE: The purpose of this study was to compare measurements between mechanical and computerized axiographs in recording the rotational and translation movements of the mandible. MATERIAL AND METHODS: The study enrolled 31 subjects with no detectable clinical signs of temporomandibular disorders. A single operator obtained 3 separate axiographic tracings of right and left condylar paths for each subject, using repeated opening, closing, protrusive, and retrusive movements. Data were collected for both the mechanical (SAM Axiograph Axo 200), and the computerized axiographs (SAM Axiotron and Axo 500). Angular (degrees) and linear measurements (mm) for the mechanical axiograph were made from the tracings obtained on grid paper using a compass, ruler, and protractor. The computer performed the measurements for the computerized axiograph tracings. All parameters were compared statistically with a Wilcoxon signed rank test (alpha = .05). The same operator remeasured all mechanical axiographic tracings for 20 subjects 1 week later to evaluate measurement error. Chronbach's alpha was used as a measure of consistency between the 2 measurements. RESULTS: Mean opening and closing angles varied between 52.4 and 54.2 degrees, and mean protrusive, and retrusive angles varied between 50.1 and 54.9 degrees. Mean opening and closing distances varied between 11.4 and 12.3 mm, and mean protrusive and retrusive distances varied between 8.2 and 8.7 mm. Comparison with the Wilcoxon signed rank test showed no significant differences for any parameter tested. There were high and medium inter-item correlations between repeated measurements of the manual device when Chronbach's alpha was applied, but these results were not sufficient to prove consistency between 2 consecutive measurements. CONCLUSION: The results showed nonsignificant differences between the recordings of the mechanical and computerized axiographs tested. The authors hypothesize that minor differences may be attributed to hand measuring errors for the tracings generated by the mechanical axiograph. This study is limited due to the impact of poor precision of the manual device on the study results when these recordings are compared to another instrument. Therefore, it is not possible to conclude that data from the manual and the computerized axiographs are compatible with each other. PMID- 16046973 TI - A technique to produce a mirror-image wax pattern of an ear using rapid prototyping technology. AB - This article describes the use of computer-aided techniques to produce a wax pattern of a missing ear. A 3-dimensional (3-D) computer model of a conventional cast from a patient was obtained using an optical surface capture device (scanner). The 3-D computer model was inverted, using computer-aided design software. A physical thermoplastic (wax) pattern of the inverted 3-D cast was produced using a rapid prototyping (RP) machine. PMID- 16046974 TI - Simple technique for cleaning and drying the barrel of a plastic impression syringe. PMID- 16046975 TI - Fabrication of a removable customized anterior guide table. PMID- 16046976 TI - A trayless impression technique for complete arch implant-supported immediately loaded provisional and definitive restorations. PMID- 16046978 TI - ASGE guideline: guideline on the use of endoscopy in the management of constipation. PMID- 16046979 TI - A prospective study comparing endoscopy and EUS in the evaluation of GI subepithelial masses. AB - BACKGROUND: The purpose of this study is to prospectively evaluate the performance characteristics of endoscopy and EUS in the diagnosis of GI subepithelial masses. METHODS: A total of 100 consecutive patients referred for the evaluation of a suspected GI subepithelial lesion were prospectively studied with endoscopy followed by EUS. Size, color, mobility, location (intramural or extramural), consistency (solid, cystic, or vascular), and presumptive diagnosis were recorded at the time of endoscopy. EUS then was performed, and size, echogenicity, location, and presumptive diagnosis were determined. RESULTS: A total of 100 subepithelial lesions were evaluated. Endoscopy had 98% sensitivity and 64% specificity in identifying intramural lesions. Size measurement by endoscopy correlated with size measurement by EUS (r = 0.88). Histology was obtained in 23 cases, with the presumptive EUS diagnosis correct in only 48% of cases. Most incorrect EUS diagnoses occurred with hypoechoic 3rd and 4th layer masses. CONCLUSIONS: Endoscopy has high sensitivity but low specificity in identifying the location (intramural or extramural) of subepithelial lesions. In addition, EUS imaging alone is insufficient to accurately diagnose 3rd and 4th layer hypoechoic masses, and histologic confirmation should be obtained whenever possible. PMID- 16046980 TI - Diagnosis of subepithelial tumors in the GI tract. Endoscopy, EUS, and histology: bronze, silver, and gold standard? PMID- 16046981 TI - A randomized single-blind trial of split-dose PEG-electrolyte solution without dietary restriction compared with whole dose PEG-electrolyte solution with dietary restriction for colonoscopy preparation. AB - BACKGROUND: Colonoscopy preparation regimens are poorly tolerated, requiring the use of a large volume of an unpalatable solution and diet restriction for adequate cleansing. The aim of this study was to compare the efficacy of two regimens of bowel preparation before colonoscopy: a whole dose of polyethylene glycol electrolyte solution (PEG-E), with diet restriction vs. a split dose with no diet restriction. METHODS: A total of 141 patients (ages 20-84 years, 81 men) were randomly assigned to receive either 4 L PEG-E, along with a liquid diet the day before colonoscopy (Group A) or 2 L PEG-E with a regular diet the day before colonoscopy followed by another 2 L PEG-E on the day of the procedure (Group B). The quality of the preparation was graded by the endoscopist (poor to excellent), who was blinded to the type of preparation. Tolerability of the assigned preparation and adverse effects were recorded by an independent investigator by using a questionnaire administered before colonoscopy. Intra- and interobserver variability was studied by using randomly chosen videotapes of colonoscopies performed as part of the study. RESULTS: There were 73 patients in Group A and 68 patients in Group B. The quality of the preparation was significantly better in Group B ( p = 0.011). The tolerability of the preparation regimen was not different overall between study groups in terms of side effects (except for bloating, which was more frequent in Group B, p = 0.039) or willingness to repeat the preparation. There was a nonsignificant trend toward improved adherence to the assigned preparation in favor of Group B ( p = 0.062). Inter- and intraobserver variability analysis showed good to excellent correlation among endoscopists. CONCLUSIONS: Colonic preparation with split-dose PEG-E and no dietary restriction provides better quality colon cleansing than whole-dose preparation, with no significant impact on patient tolerability and side effects. PMID- 16046982 TI - Women patients' preference for women physicians is a barrier to colon cancer screening. AB - BACKGROUND: The preference of women patients for women physicians has been shown in many specialties. Women patients awaiting a lower endoscopy have been shown to have a preference for women endoscopists. The reasons for this preference and the strength of this preference have not been studied in the primary care setting. METHODS: A questionnaire was given to female patients who were waiting for primary care appointments at 4 offices. Patients reported sociodemographic characteristics, experiences with colorectal cancer (CRC), barriers to CRC screening, gender preference of their physician, the significance, and reasons for this preference. RESULTS: A total of 202 women patients aged 40 to 70 years (mean 53 years) completed the questionnaire. Of these patients, 43% preferred a woman endoscopist, and of these, 87% would be willing to wait >30 days for a woman endoscopist, and 14% would be willing to pay more for one. The most common reason (in 75%) for this gender preference was embarrassment. Univariate analysis revealed that gender of the primary care physician (PCP), younger patient age, current employment, and no previous history of colonoscopy were predictors of preference for a woman endoscopist. Of these variables, only female gender of the PCP (OR 2.84: 95% CI[1.49, 5.40]) and employment (OR 2.4: 95% CI[1.23, 4.67]) were positive predictors for a woman endoscopist preference by multivariable analysis; 5% stated that they would not undergo a colonoscopy unless guaranteed a woman endoscopist. The sole independent factor associated with adherence to screening was PCP recommendation (OR 2.93: 95% CI[1.63, 5.39]). CONCLUSIONS: Women patients frequently prefer a woman endoscopist, and this preference is reported as being strong enough to delay the procedure and to incur personal expense. It is an absolute barrier to endoscopy according to 5% in this subset of women surveyed. Interventions must be made in the primary care setting to address this issue and to increase the participation of women patients in CRC screening. PMID- 16046983 TI - Safety of band ligator use in the small bowel and the colon. AB - BACKGROUND: Endoscopic band ligation for bleeding small-bowel vascular lesions has been reported as safe and efficacious based on small case series. There have been several other published case reports of band ligators used for bleeding lesions, usually Dieulafoy's lesions, in the stomach, the proximal small bowel, and the colon. In addition, this method has been used for postpolypectomy bleeding stalks. There has never been a critical look at the anatomic consequences of banding in the thinner sections of bowel. METHOD: The purpose of this study is to define the anatomic and histologic consequences of applying band ligator devices to the small and the large bowel. Fresh surgical specimens, both large and small bowel, that were excised because of neoplastic lesions were transported to our endoscopy unit where one end of the intact bowel was sutured shut. A standard upper endoscope was passed via the open end, and the bowel was closed tightly with rubber band ties. The bowel then was insufflated, and band ligators were applied to unaffected mucosa by using a standard technique. Photodocumentation from inside and outside the bowel was obtained. Some of the band polyps were cut above the band, and some were cut below the band in the fresh state. Some were fixed in formalin and examined microscopically. Histologic sectioning occurred at the level of the bands. RESULTS: The results were striking in that there were large holes (1 cm) in the fresh ileum specimen. There was gross serosal entrapment manifested by visible puckers on the outer surfaces of the specimens, especially in the small bowel and the right colon. The left colon, anatomically thicker, was less affected. The histologic evaluation revealed inclusion by the band ligator of the muscularis propria and serosa on the small bowel, the muscularis propria in the right colon, and the submucosa in the left colon. CONCLUSIONS: Based on these findings, we conclude that band ligator devices are not safe in the small bowel and the right colon but probably are safe in the thicker left colon. PMID- 16046984 TI - Endoscopic variceal ligation safety throughout the GI tract. PMID- 16046985 TI - Endoscopic suturing without extracorporeal knots: a laboratory study. AB - BACKGROUND: To eliminate cumbersome extracorporeal knotting, we designed a new endoscopic suturing device, the Eagle Claw V. The efficacy of the new device was tested on the Erlangen model and was compared with the initial extracorporeal knotting version (Eagle Claw II). METHODS: Segments of porcine splenic arteries were placed on the mucosal surface of the anterior wall of a pig stomach. The two ends of the artery were brought out through the gastric wall and were connected to a two-channel manometer. The Eagle Claw V has a curved needle with a detachable tip. After puncturing the tissue, the needle tip was engaged into a catching cartridge, where the suture had been embedded in a tightening mechanism. The suture could be tightened by simply pulling the end. The Eagle Claw V was used to plicate the splenic arteries mounted on the stomach. Suturing was considered secure if the suture could withstand endoluminal pressure greater than 200 mm Hg that lasted at least 10 seconds. RESULTS: Eleven of 15 stitches (73.3%) gained secure plication of the vessels. The suturing time (mean +/- standard deviation 2.93 +/- 0.80 minutes) was significantly faster than that of the Eagle Claw II (9.38 +/- 1.51 minutes). The device consistently achieved penetration of the muscular layer, and 4 of 15 sutures included the serosa. The 4 failures were because of suture breakage in two, thread entanglement in one, and cartridge dislodgement in one. CONCLUSIONS: The present prototype represents significant improvements in the ease of operation and the security of the suture, bringing the technique closer to clinical use for a variety of applications. PMID- 16046986 TI - Non-small-bowel lesions detected by capsule endoscopy in patients with obscure GI bleeding. AB - BACKGROUND: Approximately two thirds of patients undergoing capsule endoscopy for obscure GI bleeding will have an abnormality found in the small intestine. This report describes 9 patients (4 men, 5 women) of 140 with obscure bleeding in whom a source of their blood loss was found in the stomach or the colon at capsule endoscopy. METHODS: A review was made of a prospective database of 140 consecutive patients undergoing capsule endoscopy for obscure GI bleeding at a single center. Patients with a definite or likely cause of bleeding within reach of conventional upper or lower GI endoscopy were identified. RESULTS: Three patients had gastric antral vascular ectasia and another an inflamed pyloric canal polyp. Two patients had actively bleeding cecal carcinoma, missed at previous colonoscopies. Two others had bleeding cecal angiodysplasia. The final patient had severe nonspecific cecal inflammation. The identification of these lesions was aided by the suspected blood indicator. All patients underwent endoscopic therapy or surgery for their non-small-bowel lesions. CONCLUSIONS: Like push enteroscopy, capsule endoscopy also can identify lesions within reach of conventional endoscopy and colonoscopy. These subsequently can be treated successfully. The reasons why these lesions have been missed are unclear. PMID- 16046987 TI - Impact of EUS in the evaluation of pancreaticobiliary disorders in children. AB - BACKGROUND: While the role of EUS in the evaluation of pancreaticobiliary (PB) disorders in adults is well established, its utility in children remains unproven. This prospective study evaluates the feasibility, the safety, and the impact of EUS in the evaluation of PB disorders in children. METHODS: All children (<18 years) referred for ERCP for evaluation of suspected PB disorders who underwent EUS before scheduled ERCP. The main outcome measure was to evaluate the impact of EUS in the evaluation of PB disorders in children. EUS was considered to have a significant impact if a new diagnosis was established or if the findings altered subsequent management. RESULTS: Fourteen patients (mean age 13 years; range 5-17 years) underwent 15 EUS procedures over a 3-year period. Main indications were the following: acute or recurrent pancreatitis (6 patients), suspected biliary obstruction (5), and abdominal pain suggestive of PB origin (3). EUS diagnosed chronic pancreatitis (3 patients), idiopathic fibrosing pancreatitis (2), carcinoid tumor (1), pancreatic pseudocyst (1), pancreas divisum (1), choledocholithiasis (1), duodenal duplication cyst (1), and normal (4). Diagnosis of idiopathic fibrosing pancreatitis and carcinoid tumor was established by EUS-guided FNA. The procedure was successful in all patients, and no complications were encountered. EUS had an impact on patient management in 93% of cases: established new diagnosis (10), precluded need for ERCP (9), and provided additional information that facilitated focused endotherapy (4). A limitation was the small number of enrolled patients and absence of long-term clinical follow-up. CONCLUSIONS: EUS and EUS-guided FNA are feasible, safe, and have significant impact that alters subsequent management in the majority of children with PB disorders. Further studies and dissemination of information is required to facilitate its increased application in children. PMID- 16046988 TI - Oral allopurinol does not prevent the frequency or the severity of post-ERCP pancreatitis. AB - BACKGROUND: Pancreatitis is the most common major complication of ERCP. Efforts have been made to identify pharmacologic agents capable of reducing its incidence and severity. The aim of this trial was to determine whether prophylactic allopurinol, an inhibitor of oxygen-derived free radical production, would reduce the frequency and severity of post-ERCP pancreatitis. Methods A total of 701 patients were randomized to receive either allopurinol or placebo 4 hours and 1 hour before ERCP. A database was prospectively collected by a defined protocol on patients who underwent ERCP. Standardized criteria were used to diagnose and grade the severity of postprocedure pancreatitis. RESULTS: The groups were similar with regard to patient demographics and to patient and procedure risk factors for pancreatitis. The overall incidence of pancreatitis was 12.55%. It occurred in 46 of 355 patients in the allopurinol group (12.96%) and in 42 of 346 patients in the control group (12.14%; p = 0.52). The pancreatitis was graded mild in 7.89%, moderate in 4.51%, and severe in 0.56% of the allopurinol group, and mild in 6.94%, moderate in 4.62%, and severe in 0.58% of the control group. There was no significant difference between the groups in the frequency or the severity of pancreatitis. CONCLUSIONS: Prophylactic oral allopurinol did not reduce the frequency or the severity of post-ERCP pancreatitis. PMID- 16046989 TI - Post-ERCP pancreatitis: is allopurinol the Holy Grail? PMID- 16046990 TI - Colon polyp retrieval after cold snaring. AB - BACKGROUND: The removal of small colon polyps by cold snare transection without electrocautery effectively eliminates polyps, and anecdotal reports indicate a low risk of bleeding and perforation. Concerns about using cold snaring have centered on the risk of immediate bleeding and the difficulty in retrieving the polyp. The objective was to determine the retrieval rates of polyps after cold snaring after two different methods of resection and retrieval. METHODS: Consecutive polyps were identified by a single colonoscopist who chose the technique of polypectomy (hot snare, cold snare, or cold forceps). If cold snaring was chosen, an independent observer assigned the polyp to method A (cold resection of polyp without tenting and then suction of the transected polyp into a trap) or method B (ensnare the polyp, pull it into the colonoscope channel, and then transect it while suctioning). The size and the approximate location of all polyps were recorded and all collected specimens were sent separately for histologic examination. Results Of 519 consecutively encountered polyps, 400 were removed by cold snare: 197 were assigned to method A and 203 to method B. The mean size of polyps that were cold snared was 3.5 mm. The mean time to remove and to retrieve polyps with method A was 14.5 seconds (n = 58) and with method B was 18.1 seconds (n = 60) ( p = 0.03). There were no complications from cold snaring. The rate of successful retrieval with method A was 100% (197 of 197 polyps) and with method B was 98% (199 of 203 polyps) ( p = 0.04). CONCLUSIONS: Cold snare removal of colon polyps is associated with a high polyp retrieval rate. Each of two methods of polyp retrieval was effective. Snare transection without tenting of the polyp, followed by suctioning of the specimen off the polyp site, was more efficient, though the difference in efficiency was minimal. Difficulty or failure to retrieve polyps should not be a concern with regard to cold snare polypectomy. PMID- 16046991 TI - It ain't over 'til it's over: retrieval of polyps after colonoscopic polypectomy. PMID- 16046992 TI - Serum amylase, pancreatic stents, and pancreatitis after sphincter of Oddi manometry. AB - BACKGROUND: Serum amylase levels 2 hours after ERCP predict postprocedure pancreatitis. The value of serum amylase measurements after sphincter of Oddi manometry (SOM) and the effect of pancreatic-duct stent placement on serum amylase are unknown. METHODS: Records were reviewed for 88 SOM patients who had serum amylase measured 2 hours after the procedure. Post-SOM pancreatitis was defined as pain with a >3-fold elevation of serum amylase on the morning after SOM. "Possible pancreatitis" was defined as pain with a <3-fold elevation of serum amylase on the morning after SOM. RESULTS: Post-SOM pancreatitis and possible pancreatitis each occurred in 13% of the study cohort. Post-SOM pancreatitis was associated with the absence of a pancreatic stent and occurred in 0% of patients without a stent who had normal 2-hour serum amylase vs. 67% with elevated 2-hour serum amylase (p < 0.01). Among patients who received a stent, pancreatitis occurred in 6%, regardless of whether the 2-hour serum amylase was elevated. Possible pancreatitis occurred mainly in patients who received stents, and it also was associated with elevation of the 2-hour serum amylase. CONCLUSIONS: Elevation of the serum amylase level 2 hours after SOM predicts post-SOM pancreatitis but only in patients who do not receive a pancreatic stent. Among patients who received a stent, elevated 2-hour serum amylase levels predict subsequent findings that may be caused by attenuated pancreatitis. PMID- 16046993 TI - Eagle Claw II: A novel endosuture device that uses a curved needle for major arterial bleeding: a bench study. AB - BACKGROUND: Control of bleeding from major arteries in the upper-GI tract remains difficult with currently available endoscopic devices. We designed an endosuture device that uses a curved needle and extracorporeal knotting, and assessed the device in stopping arterial bleeding in a bench model. METHODS: Harvested porcine splenic arteries (2-mm diameter) were tunneled submucosally in pig stomach with the open end protruding into an artificially created mucosal defect. The outer end of the vessel was connected to a pulsatile pump, and red ink was infused at a pressure of about 100 mm Hg. The stomach was installed on an Erlangen endo training model. The suturing device (Eagle Claw II), mounted outside an endoscope, has an eyed, curved needle that carries a 3-0 nylon thread. After puncture, the thread was retrieved by using a hook. Three-throw square knots were tied at the thread extracorporeally and were pushed into place by using a knotting cap. The criteria of successful plication was defined as hemostasis after knotting, no leakage at pressures of >200 mm Hg, and the vessel was completely encircled by the suture. RESULTS: A total of 25 sutures were made with the mean time of 9.38 minutes (standard deviation 1.51). Control of the bleeding was obtained with 17 sutures (68%). The causes for failure were the following: a suture was too shallow (4), a loose knot (2), incorrect suture position (1), and stomach-wall edema (1). CONCLUSIONS: Control of bleeding from large arteries by using endoscopic suturing is possible. Continued refinements of the device are required. PMID- 16046994 TI - Improving the effectiveness of journal reading in gastroenterology. PMID- 16046996 TI - Emerging endoscopic options in the management of esophageal leaks (videos). PMID- 16046997 TI - Endoscopic gastrojejunostomy with survival in a porcine model. AB - BACKGROUND: We have previously reported the feasibility and the safety of an endoscopic transgastric approach to the peritoneal cavity in a porcine model. We now report successful performance of endoscopic gastrojejunostomy with survival. METHODS: All procedures were performed on 50-kg pigs, with the pigs under general anesthesia, in aseptic conditions with sterilized endoscopes and accessories. The stomach was irrigated with antibiotic solution, and a gastric incision was performed with a needle-knife and a sphincterotome. A standard upper endoscope was advanced through a sterile overtube into the peritoneal cavity. A loop of jejunum was identified, was retracted into the stomach, and was secured with sutures while using a prototype endoscopic suturing device. An incision was made into the jejunal loop with a needle-knife, and the filet-opened ends of the jejunal wall were secured to the gastric wall with a second line of sutures, completing the gastrojejunostomy. OBSERVATIONS: Two pigs survived for 2 weeks. Endoscopy and a radiographic contrast study performed after gastrojejunostomy revealed a patent anastomosis with normal-appearing gastric and jejunal mucosa. Postmortem examination demonstrated a well-healed anastomosis without infection or adhesions. CONCLUSIONS: The endoscopic transgastric approach to create a gastrojejunostomy is technically feasible and can be performed, with survival, in a porcine model. PMID- 16046998 TI - Transgastric surgery in the abdomen: the dawn of a new era? PMID- 16046999 TI - A new sinker-assisted endoscopic submucosal dissection for colorectal cancer. AB - BACKGROUND: Endoscopic submucosal dissection (ESD) is accepted as a minimally invasive treatment for early gastric cancer, but it is not widely used to treat the colorectum because of its degree of technical difficulty. Thus, a noninvasive tool that facilitates the direct visualization of the submucosal layer is needed. METHODS: Traction-assisted dissection was performed on 4 superficial lesions. The system consisted of a metallic clip attached by a nylon line to a sinker 6 x 4 x 4 mm in size and weighing 1 g. After partially dissecting the submucosa, the clip was attached to the edge of the exfoliated mucosa and the weight of the sinker pulled down the partly resected lesion. OBSERVATIONS: In all 4 cases, the sinker allowed direct visualization of the cutting line, and en bloc resection was successfully accomplished. CONCLUSIONS: These preliminary results suggest that sinker-assisted ESD is effective for the complete removal of selected superficial early colorectal cancer. PMID- 16047000 TI - Technical modification of the double-balloon endoscopy to access to the proximal side of the stenosis in the distal colon. AB - BACKGROUND: The inability to pass endoscopes beyond strictures is a considerable problem in patients with a colonic stricture. METHODS: In patients with bowel obstruction, we have modified the insertion method for double-balloon endoscopy with a long, transnasal decompression tube. OBSERVATIONS: We have succeeded in reaching the proximal side of the stricture from the oral approach across the entire small bowel in a patient. CONCLUSIONS: This modified double-balloon enteroscopy is useful for patients with bowel obstruction in whom a long decompression tube is already placed. PMID- 16047001 TI - Arterioportal fistula. PMID- 16047002 TI - Mesenteric abscess in Crohn's disease. PMID- 16047003 TI - EUS features of duodenal lesions in Henoch-Schonlein purpura. PMID- 16047004 TI - Inverted appendiceal orifice masquerading as a cecal polyp on virtual colonoscopy. PMID- 16047005 TI - A solitary rectal ulcer mimicking rectal cancer. PMID- 16047006 TI - Esophageal squamous cell carcinoma with metastasis to the ampulla. PMID- 16047007 TI - Two cases of GI involvement of Gorham's disease diagnosed by double-balloon enteroscopy: case report and review of the literature. PMID- 16047008 TI - Multiple carcinoid tumors of the ileum preoperatively diagnosed by enteroscopy with the double-balloon technique. PMID- 16047009 TI - EMR of ampullary gangliocytic paragangliomas. PMID- 16047010 TI - Reflections in front of a case of ventral hernia after PEG tube removal. PMID- 16047011 TI - Ectopic pancreas in the minor duodenal papilla presenting as upper-GI bleeding. PMID- 16047013 TI - Endoscopy or surgery for malignant GI outlet obstruction? PMID- 16047015 TI - Endoscopy or surgery for malignant GI outlet obstruction? PMID- 16047016 TI - Endoscopic sphincterotomy vs. stent placement for postcholecystectomy bile leak. PMID- 16047019 TI - Brewing a pot of hysteria. PMID- 16047021 TI - How visual salience wins the battle for awareness. PMID- 16047022 TI - The synaptic Abeta hypothesis of Alzheimer disease. PMID- 16047023 TI - Beyond migration: Dlx1 regulates interneuron differentiation. PMID- 16047024 TI - Less is more: progenitor death and cortical size. PMID- 16047025 TI - Rethinking the thalamus. PMID- 16047026 TI - Dietary polyunsaturated fatty acids in asthma- and exercise-induced bronchoconstriction. AB - Despite progress that has been made in the treatment of asthma, the prevalence and burden of this disease has continued to increase. While pharmacological treatment of asthma is usually highly effective, medications may have significant side effects or exhibit tachyphylaxis. Alternative therapies for treatment that reduce the dose requirements of pharmacological interventions would be beneficial, and could potentially reduce the public health burden of this disease. Ecological and temporal data suggest that dietary factors may have a role in recent increases in the prevalence of asthma. A possible contributing factor to the increased incidence of asthma in Western societies may be the consumption of a proinflammatory diet. In the typical Western diet, 20- to 25 fold more omega (n)-6 polyunsaturated fatty acids (PUFA) than n-3 PUFA are consumed, which promotes the release of proinflammatory arachidonic acid metabolites (leukotrienes and prostanoids). This review will analyze the evidence for the health effects of n-3 PUFA in asthma- and exercise-induced bronchoconstriction (EIB). While clinical data evaluating the effect of omega-3 fatty acid supplementation in asthma has been equivocal, it has recently been shown that fish oil supplementation, rich in n-3 PUFA, reduces airway narrowing, medication use, and proinflammatory mediator generation in nonatopic elite athletes with EIB. These findings are provocative and suggest that dietary fish oil supplementation may be a viable treatment modality and/or adjunct therapy in asthma and EIB. PMID- 16047027 TI - Effects of arginine supplementation on the humoral and innate immune response of older people. AB - OBJECTIVE: To evaluate whether oral supplementation with arginine affects the humoral and innate immune response after vaccination against Streptococcus pneumoniae in a group of people aged 60 y and older, free-living in the community. DESIGN: A randomized controlled trial with one supplemented group and one control group. SETTING: Older persons living in the community. SUBJECTS: A total of 29 adults aged 60 y and older. INTERVENTIONS: The older people were randomized into two groups, one with arginine supplementation (15 g/day) for 4 weeks after pneumococcal vaccine. The control group received only the vaccine. Anthropometric measurements and immune system function parameters: neutrophil chemotaxis and phagocytosis, natural killer cell activity, determination of serum pneumococcal polysaccharide antibodies and serum C3 and C4. RESULTS: Neutrophil phagocytosis and the serum concentration of complement (C3 and C4) did not differ between groups. IgG antibodies against pneumococcal polysaccharide serotypes 1, 5 and 6B increased in both groups. The following parameters increased in the arginine-supplemented group compared to the nonsupplemented group: neutrophil chemotaxis (34 vs 19 units of migration, P = 0.002), natural killer cell cytotoxicity (23.3 vs 13.4 10 M/Ul 40%, P = 0.011) and IgG against antigen 5 (12.3 vs 6.2 mug/ml, P = 0.044). CONCLUSIONS: This study suggests that, after the pneumococcal vaccine, the intake of arginine increased neutrophil chemotaxis, natural killer cytotoxicity and serum concentration of IgG against antigen 5 in older people. These results suggest that arginine supplementation may enhance the immune response elicited by the pneumococcal vaccine in older people. SPONSORSHIP: Supported in part by CAPES and FAEPA. PMID- 16047028 TI - The Chakaria food system study: household-level, case-control study to identify risk factor for rickets in Bangladesh. AB - OBJECTIVE: A comprehensive, multiround survey of local food systems in a rickets endemic area of Bangladesh was conducted to identify household-level risk factors for rickets. DESIGN: A household-level, case-control study was conducted in a rickets-endemic area, Chakaria, with planned comparisons between households with one or more rachitic child and neighboring households with no affected children. SETTING: A rickets-endemic area of southeastern Bangladesh, Chakaria. SUBJECTS AND METHODS: An interview-based survey was conducted in six villages in Chakaria with 199 households with at least one child showing physical signs consistent with rickets and 281 households with no affected children. RESULTS: Households with rachitic children in Chakaria had more children, more pregnant or lactating women, and fewer adults than unaffected households in that community. Affected households tended to rely on farming for their livelihood and tended to have less economic activity as indicated by less outstanding debt than their neighbors. Households with rickets were at significantly greater risk of pneumonia than were other households. Calcium undernutrition was severe and widespread in Chakaria due to a food system that offered very little of the element in accessible forms. Household diets were based on cereals and starchy vegetables. Rice and fish constituted the major source of calcium for most households, although dairy products, when used, were very important calcium sources, particularly for young children. In fact, the use of dairy products was the only household choice that led to substantial increases in the calcium intakes of children, and households that used dairy products tended to show increased calcium intakes for all of their members. CONCLUSIONS: The risk of a Chakarian household having a child with rickets appeared to be related to its economic status. Although this might be expected to be manifest as limitations in food access and/or use, rickets households failed to show a dietary pattern associated with rickets. Calcium undernutrition was prevalent and, thus, would appear to be a predisposing factor for rickets; however, calcium undernutrition was prevalent in Chakarian households with and without rickets. Therefore, it is probable that another precipitating factor(s) play a role(s) in the etiology of rickets in Chakaria. PMID- 16047029 TI - Effects of vitamin E on the toxicity of oxidized LDL on endothelial cells in vitro in smokers vs nonsmokers on diets rich in fish. AB - OBJECTIVE: To clarify whether supplementation of vitamin E can alter the low density lipoprotein (LDL) oxidation properties and thereby affect endothelial cell function and prostacyclin production in smokers compared to nonsmokers on diets rich in fish in a pilot study. DESIGN: The LDL of six smokers and six nonsmokers on habitual high fish diet was isolated before and after an 8-week supplementation of vitamin E (800 IU/day). LDL was oxidized by incubation with CuSO4. Cytotoxicity of LDL oxidized to different degrees on endothelial cells was investigated in vitro in these two groups. SETTING: Helsinki University Central Hospital; Institute of Biomedicine, Pharmacology, University of Helsinki. RESULTS: At baseline, the rate of oxidation was higher in nonsmokers than in smokers. The lag phase increased significantly after the supplementation of vitamin E both in smokers and nonsmokers. Native LDL dose dependently tended to reduce the viability of endothelial cells in vitro more markedly when isolated from smokers than from nonsmokers. Vitamin E supplementation had no beneficial effect on the cytotoxicity of oxidized LDLs in endothelial cell culture. On the other hand, simultaneous administration of Trolox, the water-soluble analogue of vitamin E, attenuated the LDL cytotoxicity on endothelial cells. The vitamin E supplementation to LDL donors attenuated the increase in prostacyclin production both in smokers and nonsmokers. CONCLUSION: Supplementation of LDL donors (healthy male volunteers on habitual fish diet) with vitamin E increased the lag phase of LDL oxidation, but, on the other hand, did not influence in vitro cytotoxicity of LDL, or prostacyclin production. PMID- 16047030 TI - Body fat measurement in adolescents: comparison of skinfold thickness equations with dual-energy X-ray absorptiometry. AB - OBJECTIVE: To compare the most commonly used equations to predict body fatness from skinfold thickness, in male and female adolescents, with dual-energy X-ray absorptiometry (DXA) as a reference method of fatness measurement. DESIGN: Cross sectional nutrition survey. SETTING: General adolescent population from Zaragoza (Spain). SUBJECTS AND METHODS: A total of 238 Caucasian adolescents (167 females and 113 males), aged 13.0-17.9 y, were recruited from 15 school groups in 11 public and private schools. The percentage fat mass (%FM) was calculated by using skinfold-thickness equations. Predicted %FM was compared with the reference %FM values, measured by DXA. The lack of agreement between methods was assessed by calculating the bias and its 95% limits of agreement. RESULTS: Most equations did not demonstrate good agreement compared with DXA. However, in male adolescents, Slaughter et al equations showed relative biases that were not dependent on body fatness and the limits of agreement were narrower than those obtained from the rest of equations. In females, Brook's equation showed nonsignificant differences against DXA and the narrowest 95% limits of agreement. Only biases from Brook and Slaughter et al equations were not dependent on body fatness in female adolescents. CONCLUSIONS: Accuracy of most of the skinfold-thickness equations for assessment of %FM in adolescents was poor at the individual level. Nevertheless, to predict %FM when a relative index of fatness is required in field or clinical studies, Slaughter et al equations may be used in adolescents from both sexes and the Brook equation in female adolescents. PMID- 16047031 TI - Commentary on "The Epidemiology of Respiratory Failure in Neonates Born at an Estimated Gestational Age of 34 Weeks or More" by Clark RH, et al. PMID- 16047032 TI - Risk factors for nosocomial infections in selected neonatal intensive care units in Colombia, South America. AB - OBJECTIVE: This study was designed to identify risk factors for nosocomial infections among infants admitted into eight neonatal intensive care units in Colombia. Knowledge of modifiable risk factors could be used to guide the design of interventions to prevent the problem. STUDY DESIGN: Data were collected prospectively from eight neonatal units. Nosocomial infection was defined as culture-proven infection diagnosed after 72 hours of hospitalization, resulting in treatment with antibiotics for >3 days. Associations were expressed as odds ratios. Logistic regression was used to adjust for potential confounders. RESULTS: From a total of 1504 eligible infants, 80 were treated for 127 episodes of nosocomial infection. Logistic regression analysis identified the combined exposure to postnatal steroids and H2-blockers, and use of oral gastric tubes for enteral nutrition as risk factors significantly associated with nosocomial infection. CONCLUSION: Nosocomial infections in Colombian neonatal intensive care units were associated with modifiable risk factors including use of postnatal steroids and H2-blockers. PMID- 16047033 TI - Neonatal lupus and IUGR following alpha-interferon therapy during pregnancy. AB - Interferon alfa-2a is a cytokine produced by recombinant DNA techniques and has antiproliferative, antiviral and immunomodulating effects. A number of case reports in the past have suggested relative safety of alpha-interferons during pregnancy with little or no effect on the fetus. A 15-year-old adolescent became pregnant while receiving alpha-interferon for essential thrombocythemia. She delivered a small-for-gestational age baby girl at 33 weeks gestation. The infant displayed a facial rash characteristic of neonatal lupus and transient thrombocytopenia; maternal and neonatal serologies were typical for drug-induced lupus. These findings suggest probable association between maternal use of alpha interferon and adverse effects in the fetus. PMID- 16047034 TI - Myelomeningocele in an infant with intrauterine exposure to efavirenz. AB - We report a case of myelomeningocele in an infant whose mother was exposed to efavirenz during the first 16 weeks of pregnancy. Although the true risk for myelomeningocele with the use of efavirenz early in pregnancy is still unknown, the findings in humans are consistent with those observed in primates and suggest that efavirenz is a potent teratogen. Thus, we suggest that efavirenz only be prescribed for women of childbearing potential when no other comparable antiretroviral options are available. PMID- 16047035 TI - Diseases of progress in neonatal care. PMID- 16047036 TI - Maternal xenoestrogen exposure and breast feeding. PMID- 16047037 TI - Erythropoietin to prevent transfusions in premature infants. PMID- 16047038 TI - Prostate cancer in US Air Force veterans of the Vietnam war. AB - US Air Force veterans of Operation Ranch Hand sprayed herbicides contaminated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in Vietnam from 1962 to 1971. Comparisons served in Southeast Asia (SEA) during the same time period but did not spray herbicides. Here we investigate a potential association between exposure to TCDD and prostate cancer. Data were available for 2516 veterans (1019 Ranch Hand and 1497 Comparison) who participated in at least one of six physical examinations starting in 1982 and had a measurement of serum TCDD. We assigned Ranch Hands to two exposure categories: Lower and Higher, based on their median 20-year cumulative TCDD level. In total, 81 Comparison and 59 Ranch Hand prostate cancers were identified between 1 January 1982 and 31 December 2003. We found no overall increase in the risk of prostate cancer in Ranch Hand veterans versus the Comparisons. There was a positive association in Ranch Hand veterans in the Higher TCDD category who served in SEA before 1969 (RR=2.27, 95% CI 1.11-4.66) when more contaminated herbicides were used, but the number of cases was small (n=15). A within-group comparison found that in Comparison veterans, time served in SEA was associated with an increased risk of prostate cancer (RR=2.18, 95% CI 1.27-3.76, >789 days versus < or =789 days). No increase in the risk of prostate cancer was observed within the Ranch Hand group in association with TCDD or time served in SEA. These analyses suggest that a longer service in SEA and exposures other than TCDD may have increased the risk of prostate cancer in Comparison veterans. PMID- 16047039 TI - Validation of self-reported proximity to agricultural crops in a case-control study of neural tube defects. AB - Self-reported perinatal exposures to chemicals or pollutant sources in case control studies of birth defects may be inaccurate due to misreporting among mothers. In a case-control study of neural tube defects delivered in California in 1987-1988, mothers of case and control infants were asked whether they lived within 0.25 mile (400 m) of agricultural crops. Responses were compared against a gold standard derived from historical agricultural land-use survey maps. The odds ratio for self-reported proximity to any crops (1.62, 95% confidence interval: 1.08, 2.43) appeared to be positively biased compared with the estimate for map based proximity (1.17, 95% confidence interval: 0.79, 1.71). This pattern was also observed for proximity to specific crops such as nonpermanent and orchard crops. For vineyards, however, we observed an increased risk associated with map based proximity (odds ratio=2.45, 95% confidence interval: 1.08, 5.58) but not with self-reported proximity (1.09, 95% confidence interval: 0.51, 2.34). The sensitivity of self-reported proximity to any crops was greater for case (65.7%) than control mothers (50.0%) while specificity was about the same for case and control mothers (87.5 vs. 89.3%), suggesting that control mothers under-reported proximity to crops. Differential reporting was also observed between geographic regions, urban and rural residents, and across levels of maternal employment and education. These results suggest differential reporting between case and control mothers as well as an influence from maternal demographic characteristics on reporting accuracy. PMID- 16047040 TI - Nitrogen dioxide prediction in Southern California using land use regression modeling: potential for environmental health analyses. AB - We modeled the intraurban distribution of nitrogen dioxide (NO(2)), a marker for traffic pollution, with land use regression, a promising new exposure classification technique. We deployed diffusion tubes to measure NO(2) levels at 39 locations in the fall of 2003 in San Diego County, CA, USA. At each sample location, we constructed circular buffers in a geographic information system and captured information on roads, traffic flow, land use, population and housing. Using multiple linear regression, we were able to predict 79% of the variation in NO(2) levels with four variables: traffic density within 40-300 m of the sampling location, traffic density within 300-1000 m, length of road within 40 m and distance to the Pacific coast. Applying this model to validation samples showed that the model predicted NO(2) levels within, on average, 2.1 p.p.b for 12 training sites initially excluded from the model. Our evaluation of this land use regression model showed that this method had excellent prediction and robustness in a North American context. These models may be useful tools in evaluating health effects of long-term exposure to traffic-related pollution. PMID- 16047041 TI - Soil ingestion in children and adults in the same family. AB - Ingestion of soil may be a potentially important pathway of exposure to environmental pollutants. Although several studies have estimated soil ingestion in children, data on ingestion in adults are sparse. The purposes of this study were to estimate soil ingestion in children aged 3 to 8 years and their parents, identify factors associated with increased ingestion, and compare ingestion rates within the same family. Food/liquid, excreta, and soil/dust samples were collected for the mother, father, and participant child for 11 consecutive days in 19 families. Soil ingestion was estimated using a mass balance approach. Soil ingestion levels in children were similar to those reported previously, whereas adult estimates were somewhat higher than previous estimates. Children's eating of dirt and parents' occupational contact with soil were associated with increased ingestion. Within families, soil ingestion levels in children and adults were not correlated, although this analysis was based on fewer than 19 participant families. Children's mean soil ingestion rates ranged from 37 to 207 mg/day depending on the tracer, with the highest values based on titanium as a tracer. Adult mean soil ingestion rates ranged from 23 to 625 mg/day depending on the tracer, with the highest value based on titanium as a tracer. Soil ingestion rate estimates were more variable in adults than in children. PMID- 16047044 TI - Antibiotic resistance: multidrug efflux proteins, a common transport mechanism? PMID- 16047045 TI - Recent developments in electrospray ionisation mass spectrometry: noncovalently bound protein complexes. PMID- 16047046 TI - Natural sesquiterpenoids. PMID- 16047047 TI - Triterpenoids. PMID- 16047048 TI - Isoflavonoids in non-leguminous families: an update. PMID- 16047049 TI - Natural guanidine derivatives. PMID- 16047050 TI - Laparoscopic live donor nephrectomy. AB - With the number of patients presently awaiting renal transplantation exceeding the number of cadaveric organs available, there is an increasing reliance on live renal donation. Of the 11,869 renal transplants performed in 2002 in the US, 52.6% were living donors from the United Network for Organ Sharing Registry. Renal allografts from living donors provide: superior immediate long-term function; require less waiting time and are more cost-effective than those from cadaveric donors. However, anticipation of postoperative pain and temporary occupational disability may dissuade many potential donors. Additionally, some recipients hesitate to accept a living donor kidney due to suffering that would be endured by the donor. It is a unique medical situation when a young, completely healthy donor undergoes a major surgical procedure to provide an organ for transplantation. It is mandatory to offer a surgical technique, which is safe and with minimal complications. It is also obvious for any organ transplantation, that the integrity of the organ remain intact, thus, enabling its successful transplantation into the recipient. An acceptably short ischemia time and adequate lengths of ureter and renal vasculature are favored. Many centers are performing laparoscopic live donor nephrectomy in an effort to ease convalescence of renal donors. This may encourage the consideration of live donation by recipients and potential donors. PMID- 16047051 TI - General surgical problems encountered in the Hajj pilgrims. AB - OBJECTIVE: The study was conducted to evaluate the pattern of general surgical admissions for future planning of staff, cost and other needs of these hospitals. METHODS: The study was conducted in 2 major hospitals in the Holy Shrine in 2 consecutive Hajj years 1423, 1424 (2003, 2004). All general surgical admissions, except those who died in the emergency room or were received dead, were included in the study. RESULTS: A total of 177 patients were admitted in both hospitals in 2 Hajj seasons. There were 139 males and 38 females with mean age of 52.7 years. Acute appendicitis and diabetic foot were the most common cause of admissions. Patients who received operative treatment totalled 87 (49.1%) and 69 (39%) were managed conservatively, while 20 (11.3%) left against medical advice. One patient was referred to higher center immediately after admission. CONCLUSION: The pilgrims are a peculiar class of patients. They should receive the best possible care but keeping in view their special needs. Furthermore, this study will help in future organization of such facilities. PMID- 16047052 TI - Post appendectomy small bowel obstruction. AB - OBJECTIVE: Postoperative small bowel obstruction is one of the adverse effects of appendectomy but its frequency varies from center to other. This study was conducted to determine the incidence of this complication among our patients who had appendectomy and identify the factors which may increase the risk. METHODS: Case notes of patients who underwent appendectomy from January 1998 to December 2003 in King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia were reviewed. Patients readmitted for adhesive small bowel obstruction were traced and their clinical data were analyzed. RESULTS: Six hundred and seven patients were eligible for the study. Six patients (1%) developed intestinal obstruction. Frequency of readmission of patients with features of intestinal obstruction ranged from 1-6 (mean of 2 times). CONCLUSION: The incidence of small bowel obstruction after appendectomy is low. The main risk factors were reviewed and measures to avoid them were suggested. PMID- 16047053 TI - Homozygous AMN mutation in hereditary selective intestinal malabsorption of vitamin B12 in Jordan. AB - OBJECTIVE: Juvenile megaloblastic anemia is a rare and often hereditary disorder of cobalamin absorption, transport or intracellular metabolism. Several syndromes present with megaloblastic anemia such as congenital megaloblastic anemia due to intrinsic factor defect and juvenile megaloblastic anemia with proteinuria due to defects in the cubilin or the amnionless protein. METHODS: We identified a large kindred with juvenile megaloblastic anemia. Four genes, GIF, CUBN, TCN1, and TCN2, was previously excluded from being responsible for the syndrome of this family who was discovered in Irbid, Jordan, during the year 1999. At that time, the amnionless (AMN) gene was not yet known to implicate in megaloblastic anemia. In this study, we screened the AMN for mutations in the Ohio State University, Iowa, United States of America. In addition, follow-up testing was carried out in the University of Iowa in 2004. RESULTS: We identified a homozygous splice site mutation in the patients. This mutation was previously detected in families from Turkey and Tunisia. It is suspected to be a founder mutation of Middle Eastern origin. CONCLUSION: Molecular testing for this specific mutation in cases of Middle Eastern origin is a valuable tool for presymptomatic diagnosis, carrier identification and perhaps prenatal diagnosis. PMID- 16047054 TI - Patterns of radiographic changes in hands and feet of rheumatoid arthritis in Saudi Arabia. AB - OBJECTIVE: The aim of the study was to characterize the pattern of radiographic changes in the hands and feet of rheumatoid arthritis in Saudi patients. METHODS: The radiographs of hands and feet of rheumatoid arthritis patients attending rheumatology outpatient clinics of King Khalid University Hospital in Riyadh, Kingdom of Saudi Arabia, over the period extending from March to June 2001, were examined and reported for the presence of osteopenia, joint space narrowing, and erosions. RESULTS: Fifty-six rheumatoid arthritis patients were studied. Their mean age was 50 + 1.9 years, and mean disease duration was 9.07 + 0.84 years. Generalized osteopenia was seen in 16/56 (29%) and periarticular osteopenia in 38/56 (68%). Joint space narrowing was present in 9/56 (16%) of feet and 35/56 (63%) of hand x-rays. Erosions were seen in 3/56 (6%) of feet and in 22/56 (39%) of hand x-rays. Significant correlation was seen between joints space narrowing, joint erosions, and disease duration. CONCLUSION: Radiographic changes in hands and feet of Saudi rheumatoid arthritis patients are less severe than those reported from the West, and the pattern is also different with less affection of the feet. PMID- 16047055 TI - Detection of Helicobacter pylori and Chlamydia pneumoniae DNA in human coronary arteries and evaluation of the results with serologic evidence of inflammation. AB - OBJECTIVE: Atherosclerosis is pathologically similar to a chronic inflammatory response. Recent reports have suggested that Chlamydia pneumoniae (C. pneumoniae) and Helicobacter pylori (H. pylori) play a role in the pathogenesis of atherosclerosis but this relation has not been confirmed on an inflammatory background. METHODS: Twenty-nine consecutive patients admitted to Suleyman Demirel University Medical School Cardiovascular Surgery Department, Isparta, Turkey between May 2002 to June 2003 were included in the study and the presence of C. pneumoniae and H. pylori DNA in atherosclerotic plaques of 14 coronary endarterectomy specimens and 15 left internal mammarian artery (LIMA) specimens as control subjects were determined by polymerase chain reaction. Serologic evidence of infection and inflammatory markers were also determined in both groups. RESULTS: Two C. pneumoniae DNA cases from the plaque group (14.3%) and 4 H. pylori DNA cases; 3 from plaque (21.4%) and one from the LIMA groups (6.7%) were detected. The C-reactive protein (mg/L) were higher in DNA positive samples of C. pneumoniae (66.58) and H. pylori (21.93) compared to DNA negatives of C. pneumoniae (8.49) and H. pylori (10.98), similarly interleukin-6 (U/L) levels were higher in DNA positive samples of C. pneumoniae (42.25) and H. pylori (56.37) compared with DNA negatives of C. pneumoniae (17.52) and H. pylori (13.28), but the differences were not statistically significant. Apolipoprotein B levels were significantly higher in C. pneumoniae immunoglobulin M positive cases (0.844 g/L) compared with negatives (0.661 g/L) (p=0.004). CONCLUSION: Chronic infections modify the serum lipid profile in a way that increases the risk of atherosclerosis. The increased titers of inflammation markers in DNA positive patients support inflammation in atherosclerosis, however, the results should be reproduced in a larger cohort. PMID- 16047056 TI - A sensitive immunoblotting method for screening of microalbuminuria in diabetic patients' urine. AB - OBJECTIVE: Urinary albumin excretion is a useful marker in the prognosis of diabetic nephropathy and microvascular diseases. Methods such as enzyme linked immunosorbent assay (ELISA), radio immunoassay(RIA), radial immunodiffusion, albu screen, micro bumin and micral test are usually used for detection and screening of microalbuminuria in these patients. With consideration to the cost of an assay, methods such as ELISA and RIA are not suitable methods for screening purpose. Therefore, the aim of this work is to set a dot immunoblotting method for the measurement and screening of microalbumin in urine samples. METHODS: The study was conducted during the period August 2001 to June 2003 at the National Research Center for Genetic Engineering and Biotechnology (NRCGEB) and Pars Hospital Laboratory of Tehran, Iran on 96 diabetic patients urine samples. First, anti human albumin antibodies (Abs) were produced in rabbit and immunoglobulin G (IgG) fraction was purified by protein-A affinity chromatography. Titer of Abs and optimum incubation conditions were tested by direct ELISA. Then different concentration of human albumin (0-300 mg/l) was loaded to nitrocellulose membranes and was assayed by dot immunoblotting method. The specificity and cross reactivity of Abs was tested by SDS-PAGE electrophoresis and western immunoblotting. The sensitivity of the method was calculated from human albumin calibration curve and compared with commercial immunoturbidimetric assays. RESULTS: Our results indicates that in using IgG with the concentrations 0.5-1 ug/ml (2 x 10-5 to 10-4 dilutions) the intensity of color directly increased with the increase of human albumin standards in blots. Western immunoblotting of urine samples did not show any cross reactivity with other urine proteins. Comparison of results of this method by commercial immunoturbidimetric methods indicates the correlation regression of approximately 0.979. The sensitivity of the method was approximately 5 mg/L of human albumin. CONCLUSION: This simple immunoblotting method could measure microalbumin in urine. This method is more suitable for screening of microalbumin in diabetic patients urine with a lower test cost. PMID- 16047057 TI - Aspirin reduces serum anti-melanocyte antibodies and soluble interleukin-2 receptors in vitiligo patients. AB - OBJECTIVE: Increased serum levels of certain immunologic markers including immunoglobulin G (IgG) anti-melanocyte/ vitiligo antibodies (V-IgG) and soluble interleukin-2 receptors (sIL-2R) are associated with augmented humoral and cellular immunity involved in melanocyte cytotoxicity during the active phase of non-segmental vitiligo. Recent reports have shown that, aspirin possesses a wide range of immunomodulatory and antioxidant properties. Therefore, the aim of the present study is to investigate the effect of long-term treatment of vitiligo patients with low-dose oral aspirin on serum V-IgG activity and sIL-2R concentration. METHODS: The present study was carried out at the Vitiligo Unit, King Abdul-Aziz University Medical Center, Jeddah, Kingdom of Saudi Arabia between March and October 2003. Eighteen female and 14 male patients with a recent onset of non-segmental vitiligo were divided into 2 equal groups. One group received a daily single dose of oral aspirin (300 mg) and the second group received only placebo for a period of 12 weeks. Serum V-IgG activity and sIL-2R concentration were determined before and at the end of treatment period. The V IgG activity was measured using cellular enzyme-linked immunosorbent assay (ELISA) following incubation of IgG antibodies with an adult cultured melanocytes. Serum sIL-2R concentration was measured using the highly sensitive quantitative sandwich ELISA utilizing a commercially available kit. RESULTS: As expected, the serum V-IgG activity and sIL-2R concentration of the active vitiligo patients (0.81 +/- 0.23 optical density (O.D.), 1428 +/- 510 pg/ml) were significantly increased compared with that of controls (0.27 +/- 0.1 O.D., 846 +/ 312 pg/ml; p<0.05, p<0.01). Aspirin-treated vitiligo patients showed significant decrease in serum V-IgG activity and sIL-2R concentration (0.32 +/- 0.08 O.D., 756 +/- 216 pg/ml) compared with that of placebo-treated patients (0.83 +/- 0.19 O.D., 1327 +/- 392 pg/ml; p<0.01). CONCLUSION: Low-dose oral aspirin treatment of active vitiligo patients can cause significant reduction in the acute serum immunologic markers of T cell activation, V-IgG activity and sIL-2R concentration with concomitant arrest of disease activity. PMID- 16047058 TI - Uterine tube-ovary relationship and fimbrial development during the fetal period. AB - OBJECTIVE: We aimed to evaluate the position, localization and development of the uterine tubes and their relationships to the ovaries. METHODS: One hundred and fifty-four uterine tubes and ovaries taken from 77 human fetuses aged between 9 40 weeks of gestation have been included in this study. The study was performed in Department of Anatomy of Suleyman Demirel University Faculty of Medicine, Isparta, Turkey in 2004. The position and localization of the uterine tubes have been determined. Consequently, uterine tube - ovary relationship and fimbrial development has been investigated. RESULTS: It has been observed that the uterine tubes are transversely and obliquely positioned. The most commonly observed position was transverse position. When the relationship between the uterine tubes and ovaries was explored, it was observed that the ovaries were localized in the superior, anterior or posterior aspect of the uterine tubes. In addition, when relationship between the obliquely positioned uterine tubes and ovaries was assessed, we found that the ovaries are located at the middle, anterior, lateral or posterior of uterine tubes. Furthermore, it indicates that the fimbrial development begins only after the 20th week and the quantity of fimbriae increased by gestational age. CONCLUSION: We conclude that the uterine tubes are not positioned as in adults during the fetal period, some variants can be seen in the uterine tube - ovary relationship and fimbrial development begins after the 20th week of gestational age and continued after birth. PMID- 16047059 TI - Bacterial skin infections among patients with hemoglobinopathies. AB - OBJECTIVE: Hemoglobinopathies are a common problem in our locality, yet there were no previous studies concerning skin lesions among those patients. We sought to find out the most common skin lesions among patients with hemoglobinopathies and the frequency of occurrence of bacterial skin infections in those patients. METHODS: A total of 168 patients with bacterial skin infections were included in this study. The specimens were collected from outpatients dermatology departments of 5 main hospitals in different areas in Basrah governorate, Iraq (hospital based study) from March 2002 to October 2002. The patients were divided into 2 groups according to the type of hemoglobin on electrophoresis. The first group included 35 patients with bacterial skin infections and hemoglobinopathies while the second one included 133 patients with normal hemoglobin. RESULTS: Recurrent bacterial skin infections was reported in 30 out of 35 patients with hemoglobinopathies (85.7%) and 45 out of 133 in those with normal hemoglobin (33.8%). In addition, Staphylococcus aureus (S. aureus) were isolated from patients with hemoglobinopathies (74.3%) and gram negative bacteria (42.9%). CONCLUSION: The study revealed that patients with hemoglobinopathies were more susceptible to recurrent bacterial skin infections than those with normal hemoglobin. In addition, S. aureus and gram negative bacteria were isolated more commonly from patients with hemoglobinopathies. PMID- 16047060 TI - An acquired form of Bernard Soulier syndrome associated with acute myeloid leukemia. AB - OBJECTIVE: To investigate glycoprotein-1b (GP-1b) expression on platelets from patients with acute myeloid leukemia (AML). METHODS: Purified platelets, obtained from AML-patients and normal control subjects, were examined for surface membrane GP1b-expression by flow cytometry and GP1b-mediated aggregation responses by aggregometry. The level of elastase in plasma from patients and controls was measured by enzymed-linked immunosorbent assay. The whole of this work was carried out at the University of Liverpool, Liverpool, United Kingdom during the period of 1994-2001. RESULTS: Platelets from the majority of AML-patients showed reduced GP1b-expression and reduced GP1b-mediated aggregation responses. Reduction in platelet GP1b-expression was associated with increased plasma elastase levels. CONCLUSION: The present study suggests that elastase, released from leukemic blasts, degrades platelet GP1b, resulting in dysfunctional circulating platelets in AML-patients. These results could explain the bleeding disorders observed in these patients. PMID- 16047061 TI - Epigenetic changes and their clinical relevance in Saudi diffuse large B-cell lymphoma. A molecular and tissue microarray analysis of 100 cases. AB - OBJECTIVE: The gene encoding the DNA repair enzyme O6-methylguanine-DNA methyltransferase (MGMT) is transcriptionally silenced by promoter hypermethylation in several human cancers including diffuse large B-cell lymphoma (DLBCL). We explored the aberrant promoter methylation of MGMT in Saudi diffuse large B-cell lymphoma and to investigate MGMT hypermethylation has an effect on patient's overall survival. METHODS: In a retrospective cohort study, 100 cases of DLBCL were collected from the Department of Pathology at King Faisal Specialist Hospital and Research Centre, Riyadh, Kingdom of Saudi Arabia. We used methylation specific polymerase chain reaction to analyze the MGMT promoter methylation status in 100 tumor DNA of Saudi DLBCL patients receiving multi drug regimens. Tissue microarray (TMA) of these cases was also constructed. The MGMT protein expression was analyzed immunohistochemically. Molecular data were correlated with clinical outcome. RESULTS: Seventy one percent (71%) of 100 DLBCL patients showed MGMT promoter hypermethylation in their lymphoma. The presence of MGMT methylation was associated with statistically significant increase in the overall survival (p=0.02). The MGMT promoter hypermethylation was independent and a strong prognostic factor. CONCLUSION: The MGMT promoter hypermethylation appears to be useful marker for predicting survival in patient with DLBCL treated with multi drug regimens including cyclophosphamide, at the same time the study shows that TMA technology is useful for immunohistochemical analysis of large lymphoma populations. PMID- 16047062 TI - Screening program for prostate cancer at a university hospital in eastern Saudi Arabia. AB - OBJECTIVE: Implementation of a pilot screening program for prostate cancer among Saudi patients that would serve as a nucleus for a Kingdom-wide screening program. METHODS: A prospective study on 1,213 Saudi males between 50-80 years of age who attended the Outpatient Department at King Fahd Hospital of King Faisal University, Al-Khobar, Kingdom of Saudi Arabia during a period of 18 months (April 2001-October 2002). They were included at random from different clinics including the urology clinic. Free and total prostate specific antigen (PSA), and digital rectal examination (DRE) of the prostate were performed in all patients. Patients with abnormal DRE or PSA were scheduled for transrectal ultrasound (TRUS) and ultrasound guided biopsy of the prostate. RESULTS: Abnormal DRE or PSA were present in 84 out of 1,213 patients. Only 63 patients agreed to have TRUS and ultrasound guided biopsies. Prostate cancer was confirmed in 14 out of 1,192 patients who completed the study (1.17%). CONCLUSION: The incidence of prostate cancer among Saudi men in this hospital based study is low. A population based screening for prostate cancer may reveal the incidence of this disease. PMID- 16047063 TI - Effects and a dose response relationship of physical activity to high density lipoprotein cholesterol and body mass index among Saudis. AB - OBJECTIVE: The objective of our study was to examine the effects and a dose response relationship of physical activity on plasma high density lipoprotein cholesterol (HDL-C) levels and body mass index (BMI) among Saudi men and women living in the metropolis of Riyadh, Kingdom of Saudi Arabia (KSA). METHODS: The sample population of 474 Saudi men and women visiting the health centers in and around Riyadh was studied from September 2003 to February 2004. The population was classified in 4 physical activity groups on the basis of frequency (F), intensity (I) and time (T) of exercise. The F, I and T value was calculated by multiplying the scores of 3 parameters. Plasma HDL-C and BMI were measured and the association of HDL-C and BMI with all 4 levels of physical activity, sedentary, light, moderate and heavy were calculated applying Pearsons correlation coefficient. RESULTS: We found that the sedentary Saudi men had mean HDL-C levels of 0.65 +/- 0.25 and BMI of 28.7 while women had 0.70 +/- 0.22 HDL-C levels and 31.06 BMI. The moderate and heavy physical activity showed an increase of 53-92% and 44.2-95% in the HDL-C levels in both men and women. The physical activity had a strong association to HDL-C and an inverse association to BMI. CONCLUSION: The substantial increase in the levels of plasma HDL-C and a moderate reduction in BMI is seen in men and women who are at moderate and heavy physical activity status, however the HDL-C level does not reach to the coronary heart disease protective level, if not combined with proper dieting. A health awareness program is urgently needed at the primary care level in KSA to educate people regarding the significance of physical activity and proper diet. PMID- 16047064 TI - Common causes of child mortality in Sana'a, Yemen. AB - OBJECTIVE: To study the common causes of child mortality at Al-Thawrah Hospital, Sana'a, Yemen. METHODS: A retrospective study of 639 children aged 0-14 years, who died in the hospital within a 4 year period. This study was carried out at the Al-Thawrah Hospital, Sana'a, Yemen, between January 2000 and December 2003. Children who were admitted to the pediatric/nursery unit and died in the hospital were included in this study. Data were collected from patient's files, hospital register and death certificate to ascertain the cause of death and their percentage. RESULTS: Out of 4575 admissions to the pediatric unit, there were 639 children deaths (13.96%). Deaths were higher among those <2 years of age. The most common causes of deaths in the pediatric unit were infections (18.1%) followed by respiratory problems (17.7%), central nervous system diseases (12%), renal (9.7%), malignancy (9.5%), shock (7.9%) and cardiac diseases(7.3%). While in the nursery, out of 4182 admissions in the same period, 823 died (19.67%), among these were pre-maturity (47%), birth asphyxia (23.7%), infections (10.9%), congenital anomalies (8.4%), central nervous system diseases (5.3%) and others (4.7%). Postmortem examination was not carried out in any of the cases. CONCLUSION: The percentage of deaths on admission among children is less than it was previously, but remains high. The majority of the causes of death are preventable. Efforts to further reduce the mortality should be directed at prevention and early treatment. PMID- 16047065 TI - Child pedestrian fatalities in Diyarbakir, Turkey. AB - OBJECTIVE: As there is an increase in modernization, transportation, unskilled, and drunk drivers, injuries from traffic accidents have taken on an epidemic form all over the world. The present study aims to describe the demographic data of childhood pedestrian fatalities in a large urban area in Turkey. METHODS: The demographic data of childhood pedestrian fatalities in a large urban area in Turkey was described retrospectively, with respect to age, gender, injury pattern and location. Cases less than 16 years of age were collected retrospectively from the files of the Branch of the Council of Forensic Medicine in Diyarbakir between 1998 and 2003. RESULTS: There were 232 childhood pedestrian fatalities among a total of 267 children involved in lethal traffic accidents. The mean age of the victims was 7.2 years, most of them being in the 6-10 years of age group (49.1%). Male predominated among our victims (76.3%). With regard to injury location, the most common site was the head (73.7%). There was an increase in pedestrian fatalities in the summer season (33.6%). There was no difference in rates of pedestrian deaths between years. CONCLUSION: Child pedestrian deaths have constituted a significant percentage of all child fatalities due to traffic accidents in our region. There were 6.4 deaths per 100,000 populations, and this rate was also higher than in other populations. PMID- 16047066 TI - Hypercalcemia syndrome. Coexisting hyperthyroidism, primary hyperparathyroidism and cancer of the gallbladder. AB - Hypercalcemia is a frequent finding in clinical practice. All possible causes must be considered in a patient with hypercalcemia. The association between both benign or malignant thyroid disease and primary hyperparathyroidism is well recognized. Up to 65% with primary hyperparathyroidism have associated thyroid abnormality. Hypercalcemia has also been associated with many malignant conditions. But, it is rarely seen in digestive tract cancer, such as carcinoma of gallbladder. Hypercalcemia syndrome is an absolutely rare entity. It is coexisting with hyperthyroidism, primary hyperparathyroidism and cancer of the gallbladder. PMID- 16047067 TI - Late presentation of a right Bochdalek hernia with a right intrathoracic stomach and organo-axial torsion. AB - A postero-lateral hernia through the foramen of Bochdalek is a rare type of congenital diaphragmatic hernia CDH. The incidence of Bochdalek hernia on the right side is 10-20% compared to the left side, and herniation of the stomach into the right pleural cavity is extremely rare. We report a case of right-sided Bochdalek hernia with a right intrathoracic stomach and organo-axial torsion misdiagnosed initially, and treated as a case of hyperactive airway disease. The child had a right thoracotomy, excision of the hernia sac that contained the stomach, greater omentum and part of the liver, reduction of the viscera into the abdominal cavity and simple closure of the diaphragmatic defect. Recovery was uneventful. This case highlights the consequences of late diagnosis and the effectiveness of surgical relief. A new clinico-anatomical classification of Bochdalek hernia is presented. PMID- 16047068 TI - Propionibacterium acnes. A cause of pneumatocele associated pneumonia. AB - Propionibacterium acnes is a normal inhabitant of the skin and mucosal surfaces and is rarely identified as a cause of significant infection. Reports of chest infections by this organism are limited. We report a case of pneumatocele associated pneumonia caused by this organism and review the literature. PMID- 16047069 TI - Small cell neuroendocrine carcinoma of the endometrium, a rare aggressive tumor. AB - This is a report of a young infertile woman with a history of 8 years amenorrhea, who presented with history of vaginal bleeding of 2 months duration. Investigations revealed a small cell neuroendocrine carcinoma of the endometrium, which penetrated half of the thickness of uterine wall. We have described the clinical progress and management of this rare and highly malignant cancer. A review of the pathological types and behavior of clear cell neuroendocrine carcinoma is presented. PMID- 16047070 TI - AIDS-related progressive multifocal leukoencephalopathy. AB - A case of progressive multifocal leukoencephalopathy associated with human immunodeficiency virus infection with a fatal outcome is presented. The disease has not been reported from our region before. The patient presented initially with hemiparesis and non-enhancing lesion on computed tomography scan that was thought to be an infarct. After a delay of 4 months, the diagnosis was made by brain biopsy. This delay probably contributed to his unfortunate outcome. The case is described, and methods of diagnosis and treatment are discussed. PMID- 16047071 TI - Cytomegalovirus infection in a patient with endogenous Cushing's syndrome. PMID- 16047072 TI - Electrocardiographic evidence of heart metastasis from a primary lung cancer. PMID- 16047073 TI - Radiofrequency sinus excision. A simple technique for sacro-coccygeal pilonidal sinus disease. PMID- 16047074 TI - Evaluation of palliative management of advanced breast cancer in Khartoum, Sudan. PMID- 16047075 TI - Effect of oral mecobalamin treatment on chest pain in patients with cobalamin deficiency and no evidence of coronary artery disease. A randomized, placebo controlled trial. PMID- 16047076 TI - Arginine, omega-3 fatty acids and nucleotide-enriched diet augment the anti inflammatory effect of diclofenac on carrageenan-induced rat paw edema. PMID- 16047077 TI - Chloramphenicol susceptible methicillin resistant Staphylococcus aureus in eastern region of Saudi Arabia. PMID- 16047078 TI - Precocious puberty revealing severe hypothyroidism. PMID- 16047079 TI - Severe digital vasospasm caused by cabergoline. PMID- 16047080 TI - Obstructive airways disease in patients with significant post- tuberculous lung scarring. PMID- 16047081 TI - Biochemical analysis of serum pancreatic amylase and lipase enzymes in patients with type 1 and type 2 diabetes mellitus. PMID- 16047082 TI - The new syndrome is not really a new syndrome. Al-Aqeel Sewairi syndrome, a new autosomal recessive disorder with multicentric osteolysis, nodulosis, and arthropathy. PMID- 16047083 TI - Study of patient-reported morbidity following V-beam pulsed-dye laser treatment of port wine stains. AB - The V-beam pulsed-dye laser (PDL) (595 nm) has gained popularity in the treatment of port wine stains (PWS). It uses longer pulse durations than the standard flashlamp-pumped pulsed-dye laser (FPDL) (585 nm) and has an in-built cooling system to protect the epidermis. This should, theoretically, reduce the treatment associated side effects, including discomfort. The aim of this questionnaire based study was to confirm the clinical impression that V-beam PDL is well tolerated. The results were compared with a historical group of 62 PWS patients treated with FPDL. Fifty-one patients took part in the current study. Only 35.7% (vs. 81% in the historical comparison group) required topical anaesthetic prior to laser treatment. A shortening in the duration of bruising (8 vs. 10 days) and of symptoms such as burning and tightness (3 vs. 10 days) was recorded. Lifestyle change after treatment was recorded by fewer patients (39 vs. 57%). We conclude that V-beam PDL is better tolerated than FPDL when used at therapeutic levels in patients with PWS. PMID- 16047084 TI - Anti-inflammatory effect of linear polarized infrared irradiation on interleukin 1beta-induced chemokine production in MH7A rheumatoid synovial cells. AB - We examined the anti-inflammatory effect of infrared linear polarized light irradiation on the MH7A rheumatoid fibroblast-like synoviocytes (FLS) stimulated with the proinflammatory cytokine interleukin (IL)-1beta. Expression of messenger ribonucleic acids (mRNAs) encoding IL-8, RANTES (regulated upon activation, normal T cell expressed and secreted), growth-related gene alpha (GROalpha), and macrophage inflammatory protein-1alpha (MIP1alpha) was measured using real-time reverse transcription polymerase chain reaction, and the secreted proteins were measured in the conditioned media using enzyme-linked immunosorbent assays. We found that irradiation with linear polarized infrared light suppressed IL-1beta induced expression of IL-8 mRNA and, correspondingly, the synthesis and release of IL-8 protein in MH7A cells. This anti-inflammatory effect was equivalent to that obtained with the glucocorticoid dexamethasone. Likewise, irradiation suppressed the IL-1beta-induced expression of RANTES and GROalpha mRNA. These results suggest that the irradiation of the areas around the articular surfaces of joints affected by rheumatoid arthritis (RA) using linear polarized light may represent a useful new approach to treatment. PMID- 16047085 TI - Pulsed Nd:YAG laser effect on eruption of rat mandibular incisors following disturbance of the enamel organ in the pulp. AB - We investigated the effects of pulsed neodymium:yttrium-aluminium-garnet (Nd:YAG) laser irradiation time on the eruption of 56 mandibular incisors in 28 rats. Clinically, some incisors erupted and the others did not. The incisors were irradiated at 2 W, 20 pulses/s for the period of 3 s, 5 s, 7 s, and 10 s, and the regeneration process was monitored at 20 days after laser treatment. Incisors irradiated for 3-5 s continued their eruption; five incisors irradiated for 7 s and all incisors irradiated for 10 s did not erupt. In the incisors that continued to erupt, the inner epithelial cells differentiated into ameloblast and a part of the pulp cavity was occupied by osteodentin. In the teeth in which eruption had ceased, the inner epithelium cells did not differentiate into ameloblast, and most of the pulp cavity was occupied by osteodentin. The results indicate that a relatively short time duration of pulsed Nd:YAG laser irradiation in the pulp induced the reparative process without disturbing the eruption. PMID- 16047086 TI - In vitro radiographic analysis of Nd:YAG-laser-irradiated dentin. AB - The aim of this study was to evaluate the effects of Nd:YAG laser irradiation on the dentin radiographic image. Previous studies have shown that Nd:YAG laser irradiation is capable of promoting morphological and chemical changes of the dentin surface, indicating the possibility of an alteration of its radiographic image. Dentin of 2-mm-thick mid-coronal sections from 68 extracted posterior teeth was irradiated for 30 s by a Nd:YAG laser (lambda=1,064 nm) on one half of its surface area. The other half was left as a control. The irradiation parameters were: 80 mJ/pulse, 0.8 W, 10 Hz, with a 320-mum flexible quartz optic fiber in contact mode. The 68 samples were radiographed with a 70-kV X-ray setup. The focus-film distance was established as 30 cm and there was no distance between the object and the film. The X-ray exposure time was set as 0.16 s and the radiographic film used was an F-speed dental film. The obtained radiographic images were submitted to a densitometric analysis. The images analyzed showed that there were statistically significant differences between the density of the irradiated dentin images and the non-irradiated dentin images. The image density data showed that the irradiated dentin radiographic images were 6.36% more radiopaque than the non-irradiated dentin images. The Nd:YAG laser irradiation of dentin is able to alter its radiographic image, producing more radiopaque images of the irradiated dentin. PMID- 16047087 TI - Psychosocial predictors of cannabis use in adolescents at risk. AB - This research has tested a social disintegration model in conjunction with risk and protection factors that have the power to differentiate relative, weighted interactions among variables in different socially disintegrated groups. The model was tested in a cross-sectional sample of 1082 at-risk youth in Switzerland. Structural equation analyses show significant differences between the social disintegration (low, moderate, high) groups and gender, indicating that the model works differently for groups and for gender. For the highly disintegrated adolescents results clearly show that the risk factors (negative mood, peer network, delinquency) are more important than the protective factors (family relations, secure sense of self). Family relations lose all protective value against negative peer influence, but personal variables, such as secure self, gain protective power. PMID- 16047088 TI - Assessing the effects of the dating violence prevention program "safe dates" using random coefficient regression modeling. AB - The Safe Dates Project is a randomized trial for evaluating a school-based adolescent dating violence prevention program. Five waves of data were used to examine the effects of Safe Dates over time including primary and secondary prevention effects, moderators, and mediators of program effects. Using random coefficients models, with multiple imputation of missing data, significant program effects were found at all four follow-up periods on psychological, moderate physical, and sexual dating violence perpetration and moderate physical dating violence victimization. Marginal effects were found on sexual victimization. Effects on severe physical perpetration at all four follow-up periods were moderated by prior involvement in that type of violence. Primary and secondary prevention effects were found and the program was equally effective for males and females and for whites and non-whites. Program effects were mediated by changes in dating violence norms, gender-role norms, and awareness of community services. PMID- 16047089 TI - Genetic counseling for fragile x syndrome: updated recommendations of the national society of genetic counselors. AB - These recommendations describe the minimum standard criteria for genetic counseling and testing of individuals and families with fragile X syndrome, as well as carriers and potential carriers of a fragile X mutation. The original guidelines (published in 2000) have been revised, replacing a stratified pre- and full mutation model of fragile X syndrome with one based on a continuum of gene effects across the full spectrum of FMR1 CGG trinucleotide repeat expansion. This document reviews the molecular genetics of fragile X syndrome, clinical phenotype (including the spectrum of premature ovarian failure and fragile X-associated tremor-ataxia syndrome), indications for genetic testing and interpretation of results, risks of transmission, family planning options, psychosocial issues, and references for professional and patient resources. These recommendations are the opinions of a multicenter working group of genetic counselors with expertise in fragile X syndrome genetic counseling, and they are based on clinical experience, review of pertinent English language articles, and reports of expert committees. These recommendations should not be construed as dictating an exclusive course of management, nor does use of such recommendations guarantee a particular outcome. The professional judgment of a health care provider, familiar with the facts and circumstances of a specific case, will always supersede these recommendations. PMID- 16047090 TI - Policy recommendations for carrier testing and predictive testing in childhood: a distinction that makes a real difference. AB - The genetic testing of children raises many ethical concerns. This paper examines how five position statements from Canada, UK and USA, which present guidelines for good practice in this area produce different recommendations for carrier testing and predictive testing. We find that the genetic information generated through carrier testing is routinely presented as less serious than that generated from predictive testing. Additionally, the reproductive implications of predictive testing are also routinely erased. Consequently, the papers argue strongly against predictive testing but advise caution against carrier testing in somewhat weaker terms. We argue that these differences rest on assumptions about the status of reproduction in people's lives and on an ethical stance that foregrounds the self over others. We propose that questioning the crude and sharp distinction between carrier and predictive testing in principle may enable practitioners and parents/families to make more nuanced decisions in practice. PMID- 16047091 TI - Educating genetic counselors in Australia: developing an international perspective. AB - The demand for genetic counseling services is increasing worldwide. This paper highlights the Australian experience of genetic counselor education and the history of the profession. The relevance of local factors, including the health care system, the education system and the small population in the evolution of the 1-year training programs are considered as an alternative model for emerging programs. The development of the education and training processes compared to that of other countries namely the United States of America (USA), the United Kingdom (UK) and Canada is discussed. The importance of international collaborations between the programs, to facilitate academic discussion and possible curriculum innovations, and to maintain professional understanding between genetic counselors is emphasized. Core genetic counseling competencies have been published for the UK and USA and an Australian set is proposed. In conclusion future directions are considered, including international issues around genetic counseling certification, reciprocity, and the potential for an Australian role in training genetic counselors in South East Asia. PMID- 16047092 TI - Women's attitudes toward testing for fragile X carrier status: a qualitative analysis. AB - Fragile X syndrome (FXS) is primarily due to a repeat expansion mutation found in the FMR1 X-linked gene. We have conducted a qualitative analysis of responses from women concerning their attitudes toward testing for carrier status of the fragile X mutation among reproductive-age women. We obtained responses from focus groups including women with and without FXS in their families. We found the following themes: (1) mothers of children with FXS have difficulty formulating their opinions on population screening because of their unique experiences surrounding their own carrier diagnosis and their relationship with their children with FXS, (2) the motivation for carrier testing and need for information differ by family history of FXS and parental status, and (3) the timing of carrier testing with respect to a woman's life stage dictates whether carrier information will be viewed as beneficial or detrimental. There was evidence that non-carrier women from the general population would be wholly unprepared for positive carrier results. These findings have significant implications for genetic counseling as well as for population screening. PMID- 16047093 TI - Psychosocial issues that face patients with Charcot-Marie-Tooth disease: the role of genetic counseling. AB - Charcot-Marie-Tooth (CMT) disease is a hereditary debilitating progressive muscular atrophy and sensory neuropathy of the distal extremities. CMT is usually non-life threatening. Signs of the disease usually present in childhood or in young adulthood and the level of disability can be variable within and between families. Research addressing specific psychosocial and emotional issues faced by individuals with CMT is limited. Fourteen adults with a clinical and/or molecular diagnosis of CMT (ages 32--74 years) consented to an audio taped interview. The format of the interview was based around an informal questionnaire to prompt and guide the interviewee to describe their experiences of living with a disabling genetic disorder. The interviews focused on their experiences of first symptoms and diagnosis, their life experience with CMT, their limitations due to disability and the role of genetic counseling. This study identifies and explores life issues that individuals with CMT may face, specifically grief over the loss of independence, emotional pain and stress such as embarrassment and guilt of passing on a gene mutation, impact on quality of life, the impact of wearing orthopedic devices, and fear of progressive disability. Our findings suggest that that there are emotional and psychosocial issues specific to affected individuals at different life stages and genetic counselors need to be aware of these issues in order to provide age appropriate support and advice to individuals affected by CMT. PMID- 16047094 TI - An assessment of risk understanding in Hispanic genetic counseling patients. AB - This study sought to identify if differences existed in risk comprehension and risk format understanding between genetic counseling patients of Hispanic and Caucasian ethnicity. A total of 107 questionnaires were collected, 56 from Hispanic patients, and 51 from Caucasian controls. Of the total population 41.1% (44/107) could not demonstrate sufficient risk understanding, which was 71.4% (40/56) of Hispanics and 7.8% (4/51) of Caucasians. Fractions were the best understood format for all participants. However, both Hispanics and Caucasians had difficulties with the percentage risk format. Discrepancies were also noted in qualitative word format understanding. Awareness of differences in risk comprehension may affect the selection of counseling techniques and strategies utilized by genetic counselors when educating patients about risk related information. PMID- 16047095 TI - Nitrogen turnover in drying sediments of an Amazon floodplain lake. AB - In the Amazon floodplain large areas are subject to annual cycles of drying and rewetting. The turnover of nitrogen in the periodically drying sediments is an important regulator of floodplain fertility. In the present study the transition of a lake sediment from flooded to dry conditions was studied with respect to microbial nitrogen turnover. Soil nitrogen pools, as well as the activity and abundance of denitrifying and nitrifying bacteria, were investigated during one dry season. During the first weeks after drying, most of the inorganic nitrogen vanished from the sediment. The process was inhibited by a nitrification inhibitor, showing that coupled nitrification-denitrification was responsible for the nitrogen loss. Assimilation by plants or microbes, as well as leaching, were not important mechanisms of nitrogen loss. During a period of only 10 days, 59% of the total denitrification and 94% of the total N2O emission during the dry period occurred. Cell numbers of denitrifiers were not correlated with activities. Denitrification was not correlated with other sediment variables but was regulated by the patchy distribution of reduced and oxidized zones in the uppermost centimeters of the sediment. This heterogeneity was probably introduced by the bioturbation of small insects, which was restricted to a rather short time period shortly after drying. PMID- 16047096 TI - Isolation of iodide-oxidizing bacteria from iodide-rich natural gas brines and seawaters. AB - Iodide-oxidizing bacteria (IOB), which oxidize iodide (I-) to molecular iodine (I2), were isolated from iodide-rich (63 microM to 1.2 mM) natural gas brine waters collected from several locations. Agar media containing iodide and starch were prepared, and brine waters were spread directly on the media. The IOB, which appeared as purple colonies, were obtained from 28 of the 44 brine waters. The population sizes of IOB in the brines were 10(2) to 10(5) colony-forming units (CFU) mL(-1). However, IOB were not detected in natural seawaters and terrestrial soils (fewer than 10 CFU mL(-1) and 10(2) CFU g wet weight of soils(-1), respectively). Interestingly, after the enrichment with 1 mM iodide, IOB were found in 6 of the 8 seawaters with population sizes of 10(3) to 10(5) CFU mL(-1). 16S rDNA sequencing and phylogenetic analyses showed that the IOB strains are divided into two groups within the alpha-subclass of the Proteobacteria. One of the groups was phylogenetically most closely related to Roseovarius tolerans with sequence similarities between 94% and 98%. The other group was most closely related to Rhodothalassium salexigens, although the sequence similarities were relatively low (89% to 91%). The iodide-oxidizing reaction by IOB was mediated by an extracellular enzyme protein that requires oxygen. Radiotracer experiments showed that IOB produce not only I2 but also volatile organic iodine, which were identified as diiodomethane (CH2I2) and chloroiodomethane (CH2ClI). These results indicate that at least two types of IOB are distributed in the environment, and that they are preferentially isolated in environments in which iodide levels are very high. It is possible that IOB oxidize iodide in the natural environment, and they could significantly contribute to the biogeochemical cycling of iodine. PMID- 16047097 TI - Design, development, and use of molecular primers and probes for the detection of Gluconacetobacter species in the pink sugarcane mealybug. AB - Molecular tools for the species-specific detection of Gluconacetobacter sacchari, Gluconacetobacter diazotrophicus, and Gluconacetobacter liquefaciens from the pink sugarcane mealybug (PSMB) Saccharicoccus sacchari Cockerell (Homiptera: Pseudococcidae) were developed and used in polymerase chain reactions (PCR) and in fluorescence in situ hybridizations (FISH) to better understand the microbial diversity and the numerical significance of the acetic acid bacteria in the PSMB microenvironment. The presence of these species in the PSMB occurred over a wide range of sites, but not in all sites in sugarcane-growing areas of Queensland, Australia, and was variable over time. Molecular probes for use in FISH were also designed for the three acetic acid bacterial species, and shown to be specific only for the target species. Use of these probes in FISH of "squashed" whole mealybugs indicated that these acetic acid bacteria species represent only a small proportion of the microbial population of the PSMB. Despite the detection of Glac. sacchari, Glac. diazotrophicus, and Glac. liquefaciens by PCR from different mealybugs isolated at various times and from various sugarcane-growing areas in Queensland, Australia, these bacteria do not appear to be significant commensals in the PSMB environment. PMID- 16047098 TI - Screening for putative PGPR to improve establishment of the symbiosis Lactarius deliciosus-Pinus sp. AB - A screening for plant growth promoting rhizobacteria (PGPR) was carried out in the mycorrhizosphere of wild populations of Pinus pinea and P. pinaster, and in the mycosphere of associated Lactarius deliciosus. A total of 720 bacteria were isolated, purified, and grouped by morphological criteria. Fifty percent of the isolates were selected and tested for aminocyclopropanecarboxylic acid (ACC) degradation, auxin and siderophore production, and phosphate solubilization. Thirty eight percent of the isolates showed at least one of the evaluated activities. Nutrient-related traits were associated with P. pinaster, whereas hormone production traits predominated in P. pinea. These activities were found mostly in Gram positive isolates. After PCR-RAPDs (random amplified polymorphic DNA) analysis, 10 groups appeared with 85% similiarity when considering all isolates, indicating the low diversity in the system. One strain of each group was identified by 16S rDNA sequencing. Our results suggest that P. pinaster selects for mycorrhizosphere bacteria that mobilize nutrients, whereas P. pinea selects for bacteria that have the capacity to increase root growth via production of plant growth regulators. PMID- 16047099 TI - Genetic diversity of Frankia microsymbionts from the relict species Myrica faya (Ait.) and Myrica rivas-martinezii (S.) in Canary Islands and Hawaii. AB - In the Western Canary Islands, Myrica faya and Myrica rivas-martinezii (Myricaceae) are phylogenetically close, endemic, actinorhizal species presumed to be remnants either of the European or the African Tertiary floras. Unisolated Frankia strains from field-collected nodules on Tenerife, Gomera, and La Palma Islands were compared by their rrs gene and 16S-23S intergenic spacer (IGS) restriction patterns. To compare the genetic diversity of Frankia strains from within and outside the host's native range, nodules of M. faya field plants were collected both in Canary Islands and in Hawaii, where this species is an exotic invasive. Myrica rivas-martinezii, endemic to the Canary Islands, was sparsely nodulated in the field. Frankia strains harbored in field-collected nodules of M. faya and M. rivas-martinezii belonged to the Elaeagnaceae strains' genetic cluster and exhibited a high degree of diversity. Frankia genotypes were specific to each host species. In the Canary archipelago, we found no relationship between site of collection and Frankia genotype for M. faya. The only exceptions were strains from site 2 in Tenerife, a location with a geological history different from the other sites sampled. Hawaiian and Canarian M. faya strains had no genotypes in common, raising questions concerning the origin of M. faya-infective Frankia in Hawaii. Nodular strains of M. rivas-martinezii from nursery plants were genetically characterized and shown to be divergent from the strains of field-collected nodules and belong to the Alnus-Casuarina strains cluster. This suggests Myrica may have the potential to nodulate with a broader range of Frankia genotypes under artificial conditions than has been detected in field collected nodules. PMID- 16047102 TI - Around PediHeart: bicuspid aortic valves and aortic dissection. PMID- 16047103 TI - Around PediHeart: chemoprophylaxis (CPX) against endocarditis. PMID- 16047104 TI - Around PediHeart: Cardiac transplantation. PMID- 16047105 TI - Multiple intracranial subarachnoid osteomas. AB - OBJECTIVE AND IMPORTANCE: Intracranial osteomas, which have no connections with the dura or the skull, are very rare. Here we report one case of multiple intracranial subarachnoid osteomas. CLINICAL DESCRIPTION: A 24-year-old gentleman, who had no previous history of systemic disease, meningitis or head injury, presented with a 6-month history of non-specific intermittent headache over the whole head. Neuro-imaging showed multiple ossified lesions in the right frontal area, which were found to compress the underlying cerebral hemisphere. The patient underwent a right frontal craniotomy. The masses were dissected from the depressed brain along the intact pial planes, however, all of the white bone hard masses tightly adhered to the superficial cortical veins. The masses were removed and the cortical veins were sacrificed. The patient had headaches and was nauseous for three days after the surgery due to venous congestion; however, the patient was neurologically intact on discharge. Pathological diagnosis was osteoma. CONCLUSION: To our knowledge, this is the first report of multiple intracranial subarachnoid osteomas. Surgical findings strongly support the hypothesis of the origin of the tumour that the primitive mesenchymal cells from the connective tissue might have migrated into the subarachnoid space accompanying the intracerebral blood vessels. PMID- 16047106 TI - Risk factors of cerebrospinal fluid rhinorrhea following transsphenoidal surgery. AB - OBJECTIVE: Cerebrospinal fluid (CSF) rhinorrhea is a potentially life-threatening complication following transsphenoidal surgery (TSS). METHODS: To elucidate the risk factors that may affect the incidence of postoperative CSF rhinorrhea, we retrospectively reviewed 200 consecutive cases of TSS performed by a single surgeon for 168 adenomas and 32 other sellar and parasellar lesions. RESULTS: Intra-operative CSF leakage was encountered in 38 cases (19.0%). Its incidence did not correlate to tumor size. Among 4 microadenoma cases with CSF leak, 3 were ACTH adenomas. In contrast, postoperative CSF rhinorrhea was observed in 5 cases (2.5%), all following TSS for adenomas. It was frequently noted in cases with prior TSS (3/40, 7.5%, p = 0.0235) and prior radiotherapy (2/7, 28.6%, p < 0.0001). Two cases who required surgical intervention had received TSS and radiotherapy previously. CONCLUSION: The risk of postoperative CSF rhinorrhea is significantly increased in cases with prior TSS or radiotherapy or both. In addition, these cases tended to show delayed CSF leaks and require sellar reconstruction for its treatment. When a CSF leak was encountered during TSS in these high-risk cases, thorough sellar reconstruction and long-term follow-up is necessary. PMID- 16047107 TI - 3-Dimensional computed tomographic angiography for use of surgery planning in patients with intracranial aneurysms. AB - BACKGROUND: After subarachnoid haemorrhage (SAH) diagnostic evaluation of the underlying cause is warranted since the rebleeding rate is high. The objective of the study was to answer the question, whether 3-Dimensional computed tomographic angiography (3D-CTA) is able to accurately determine the surgical indications in patients with intracranial aneurysms. METHODS: After performing 3D-CTA the size of the aneurysm, direction of the aneurysmal dome, neck position and variants of the circle of Willis were analysed. Surgery was performed solely on CTA data in those cases, where the aneurysm was clearly visible. If the findings were negative or inconclusive, intra-arterial digital subtraction angiography (DSA) was also done. FINDINGS: Between January 2001 and December 2002 100 patients (68 F, 32 M) were examined and 123 aneurysms (86 ruptured and 37 unruptured) were diagnosed. All patients received CTA preoperatively and in 27 patients selective DSA was additionally performed. Postoperatively in 34 patients the operative result was checked by DSA. A good correlation between CTA and the intra-operative findings was present in 92 of 100 patients. One aneurysm was not seen on CTA, but was on DSA. In four cases we could confirm DSA findings in CTA after re evaluation of the data. In three cases neither CTA nor DSA clearly showed an aneurysm, but it was confirmed during surgery. A good correlation between CTA and DSA was found in 60 of 61 patients (98%). The correlation between CTA and intra operative findings was good as expected in 92 patients, in 5 patients an aneurysm was detected on re-evaluation. Only one aneurysm could not be demonstrated by CTA but in DSA. CONCLUSION: CTA is less invasive, less time consuming, cheaper and easier to demonstrate the essential information regarding the aneurysm than DSA. We therefore recommend that following a careful analysis most aneurysms - 92% - can be operated solely on CTA data. PMID- 16047108 TI - The prognostic value of clinical characteristics and parameters of cerebrospinal fluid hydrodynamics in shunting for idiopathic normal pressure hydrocephalus. AB - BACKGROUND: It is difficult to predict which patients with symptoms and radiological signs of normal pressure hydrocephalus (NPH) will benefit from a shunting procedure and which patients will not. Risk of this procedure is also higher in patients with NPH than in the overall population of hydrocephalic patients. The aim of this study is to investigate which clinical characteristics, CT parameters and parameters of cerebrospinal fluid dynamics could predict improvement after shunting. METHODS: Eighty-three consecutive patients with symptoms and radiological signs of NPH were included in a prospective study. Parameters of the cerebrospinal fluid dynamics were measured by calculation of computerised data obtained by a constant-flow lumbar infusion test. Sixty-six patients considered candidates for surgery were treated with a medium-pressure Spitz-Holter valve; in seventeen patients a shunting procedure was not considered indicated. Clinical and radiological follow-up was performed for at least one year postoperatively. FINDINGS: The odds ratio, the sensitivity and specificity as well as the positive and negative predictive value of individual and combinations of measured parameters did not show a statistically significant relation to clinical improvement after shunting. CONCLUSIONS: We conclude that neither individual parameters nor combinations of measured parameters show any statistically significant relation to clinical improvement following shunting procedures in patients suspected of NPH. We suggest restricting the term normal pressure hydrocephalus to cases that improve after shunting and using the term normal pressure hydrocephalus syndrome for patients suspected of NPH and for patients not improving after implantation of a proven well-functioning shunt. PMID- 16047109 TI - Multitracer PET imaging in Heidenhain variant of Creutzfeldt-Jakob disease. PMID- 16047110 TI - Conservative treatments of children with episodic tension-type headache. A systematic review. AB - OBJECTIVE: We aimed to assess the effectiveness of conservative treatments in children with tension-type headaches (TTH). METHOD: We searched Medline, Embase and the Cochrane Library and performed reference checking. Two independent authors assessed trial quality and extracted data. Analysis was carried out according to type of intervention. RESULTS: In total we included 11 trials (427 TTH patients, age between 7-18 years) in this review. None of the studies were considered to be of high quality. Eight studies evaluated the effectiveness of relaxation training. Four studies included a waiting list or no treatment control group and in one study a placebo was used. CONCLUSION: We found conflicting evidence about the effectiveness of relaxation therapy when compared with no treatment or other treatments, with Relative Risks varying from 1.4 (0.6-3.2) to 10.6 (1.6-71.3). With regard to all other interventions evaluated (such as biofeedback, cognitive therapy or flupirtin) no evidence is found for or against their effectiveness. PMID- 16047111 TI - Corticospinal involvement in patients with a portosystemic shunt due to liver cirrhosis: a MEP study. AB - Hepatic myelopathy (HM) is a rare complication of chronic liver diseases usually associated with a portosystemic shunt, causing a progressive spastic paraparesis, and is likely to be overlooked. Thirteen patients with liver cirrhosis associated with surgical or spontaneous portosystemic shunts were studied to determine the frequency and gravity of HM. Six patients exhibited clear-cut signs of spinal cord involvement and four of them exhibited varying degrees of disability. Neurological examination did not reveal any abnormalities in the other patients. Motor evoked potentials (MEPs) were measured in all patients; in five of them the examinations were done before and after orthotopic liver transplantation (OLT). The patients with clinical signs of spinal cord involvement exhibited severe neurophysiological abnormalities, whereas milder but unequivocal MEP abnormalities were found in four of the seven patients with normal clinical examination. The clinical and neurophysiological features of patients with slight MEP abnormalities improved after OLT, whereas the patients with a more advanced stage of disease (severe MEPs abnormalities) did not. Our findings indicate that MEP studies may disclose an impairment of the corticospinal pathways even before HM is clinically manifest and provide evidence that early diagnosis of HM and subsequent immediate liver transplantation have to be recommended. PMID- 16047112 TI - HIV-associated neurosyphilis mimicking acoustic neurinoma. PMID- 16047113 TI - Association study of the G258S transferrin gene polymorphism and Parkinson's disease in the Spanish population. PMID- 16047114 TI - Case of tetralogy of Fallot associated with hypoplasia of the right pulmonary artery. The right internal mammary artery as an aortopulmonary collateral artery in tetralogy of Fallot. AB - A three-month-old girl was referred to pediatric cardiology unit for evaluation of mild cyanosis and murmur. Chest roentgenogram revealed a boot-shaped heart. Echocardiography demonstrated subaortic ventricular septal defect, aortic override and infundibular stenosis. Then, the patient diagnosed as tetralogy of Fallot (TOF). Cardiac catheterization and angiocardiography revealed infundibular narrowing, hypoplasia of the right pulmonary artery and the enlarged right internal mammary artery originating from the right subclavian artery supplied blood flow into the right lung. The patient was performed complete repair. Post operative period was uneventful. This is the first report presented about a case which pulmonary blood flow is provided by the enlarged right internal mammary artery in TOF associated with hypoplasia of the right pulmonary artery. PMID- 16047115 TI - Visualization of aorto pulmonary connections in tetralogy of Fallot. PMID- 16047116 TI - Percutaneous coronary stenting in guide-induced aortocoronary dissection: angiographic and CT findings. AB - We report here on a case of accidental aortocoronary dissection that occurred during the engagement of a guiding catheter. This resulted in an antegrade dissection into the right coronary artery, and a retrograde extension of the dissection into the Sinus of Valsalva and the ascending aorta up to the aortic arch. It was successfully treated with a stent deployment at the RCA ostium; this restored optimal coronary blood flow and there was a complete resolution of the aortic dissection as was documented by coronary angiography and the follow-up CT scan. PMID- 16047117 TI - 'Natural Conduit' between two atria associated with atrial septal defect. AB - Herein, we present a case of 46 years old woman with an extracardiac venous connection between two atria associated with secundum type atrial septal defect (ASD). This interatrial tunnel was not recognized by both transthoracic and transesophageal echocardiographic examinations. Computed tomography suspected a blood flow from right inferior pulmonary vein to inferior caval vein. Consequently, the patient underwent cardiac catheterization. Angiography revealed a tunnel between left and right atrium. To our knowledge, this is the first report of a case with venous tunnel between two atria associated with secundum type ASD in the literature. PMID- 16047118 TI - Aortico-right atrial tunnel in an adult patient. AB - We describe an asymptomatic 27-year-old man who presented a continuous murmur, and was found to have a tunnel between the aorta and the right atrium. Successful surgical treatment was performed. We emphasize the rarity of this condition, and discuss its major features. PMID- 16047119 TI - An uncommon coronary artery fistula causing survived sudden cardiac death in a young woman. AB - Although most patients suffering from a coronary artery fistula remain asymptomatic during childhood and adolescence, many of them develop complications in adulthood due to volume overload, endocarditis or ischemia. Here we present a young woman surviving a sudden cardiac death due to myocardial ischemia with subsequent ventricular fibrillation caused by thrombosis of a coronary artery fistula deriving from the left main coronary artery. Parts of the thrombus had been embolized into the circumflex artery causing posterior myocardial infarction. The thrombosis might have been initiated by local compression from a round extracardiac mass of 3 cm with liquid and solid content located between the vena cava superior and the ascending aorta. This structure was suspected to be a hematoma which had been caused by a blunt thoracic trauma while playing soccer 2 weeks before. PMID- 16047120 TI - Intracardiac echocardiography: gross anatomy and magnetic resonance correlations and validations. AB - PURPOSE: The feasibility and safety of intracardiac echocardiography (ICE) in humans, using low frequency transducers, and its excellent tissue contrast capabilities that enhances the differentiation of intracardiac structures have been previously demonstrated. However, correlations among ICE imaging and anatomic sections or magnetic resonance (MR) scan planes have never been described before. This study was designed to correlate a simplified ICE approach with the anatomy of the right atrium and great vessels obtained by serial post mortem sections and cardiac MR images. METHODS: A stepwise approach to ICE, which is based on our experience on over 300 consecutive patients with interatrial communications, has been correlated with anatomic sections from pressure-perfused fixed hearts and spin echo cardiac MR imaging. A 9F-9 MHz mechanical device was used to record four transverse and one longitudinal sections for an extensive evaluation of the intracardiac architecture. RESULTS: ICE transverse and longitudinal views allowed the detection of all the required information. Moreover, the anatomic sections and cardiac MR scan planes allowed validation of all the structures imaged by ICE. The potential clinical applications of the ICE technique are further discussed. CONCLUSION: This paper demonstrates the basic accuracy of this new imaging modality. Therefore, mechanical ICE might be considered an active investment in cardiac catheterization laboratories, specifically in the percutaneous interventional procedure setting, discovering a new route (let's see and treat) to sophisticated interventions. In this respect, there can be little doubt that the knowledge of cross-sectional mechanical ICE imaging provides the basis for understanding the heart anatomy. PMID- 16047121 TI - Intracardiac echocardiography. A new tool in the cath lab. PMID- 16047122 TI - The prospective clinical and scintigraphic assessment of patients with preserved left ventricular systolic function after transmyocardial laser revascularisation. AB - AIM: To analyse the clinical outcome and myocardial perfusion and function after transmyocardial revascularisation (TMR) in patients with normal left ventricular function and multivessel coronary artery disease non-amenable for standard revascularisation. METHOD AND RESULTS: Twenty three severely symptomatic patients (CCS score median 4) with normal left ventricular systolic function but coronaries non-amenable for either PTCA or CABG were subjected to TMR. The angina score, left ventricular systolic and diastolic function in radionuclide ventriculography at rest, exercise tolerance and myocardial perfusion--Thallium 201 SPECT (adenosine stress 74 and 37 MBq under nitrate cover) were evaluated before and 3, 6, 12 months post-operatively. After an average of 40 +/- 12 (range 14-56) TMR channels angina score decreased significantly (p< 0.0001) and the exercise tolerance increased (from 6.0 +/- 4.5 to 9.1 +/- 4.6 after 6 months, p< 0.05) in 21 patients. During the follow up period two patients had a myocardial infarction and one committed suicide after 6 months. Ejection fraction dropped significantly only after 1 year post-TMR from 70 +/- 13 to 63 +/- 0.13%, p < 0.05. The overall perfusion improved initially in 14 patients with subsequent deterioration in time. The changes in segmental perfusion were not associated with the symptomatic improvement. CONCLUSION: Transmyocardial revascularisation in patients with normal ejection fraction may improve the angina class, exercise tolerance and overall but not segmental perfusion and does not show any immediate effect on left ventricular function. PMID- 16047123 TI - Use of atropine in patients with recent myocardial infarction during exercise myocardial perfusion study. Use of atropine during exercise myocardial perfusion SPECT. AB - BACKGROUND: Patients with recent myocardial infarction frequently require a myocardial perfusion study for risk stratification. However a conclusive study cannot be accomplished in many due to non-attainment of target heart rate. AIM OF THE STUDY: To evaluate the effect of pre-stress administration of atropine in exercise performance of patients with recent myocardial infarction. MATERIAL AND METHODS: Study included 43 test and 43 control patients and were matched for their age, sex, status of pretest exercise tolerance, area of infarction and for interval between infarction and stress thallium test. Atropine (0.01 mg/kg) was administered intravenously 3 min prior to treadmill stress in all the 43 test patients and no atropine was administered to control patients. RESULTS: Target heart rate was attained in 95.3% patients receiving atropine compared to in 67.4% of control patients. There was no significant difference between mean ages (p>0.33), basal BP (p>0.47), peak BP (p>0.18) of both groups. There was significant difference between the increment in exercise-induced heart rate (p<0.004), peak heart rate (p<0.001) and double product (p<0.001) attained between both groups. No significant adverse effect was noted in patients who received atropine. CONCLUSION: Pre-stress administration of atropine in patients with recent myocardial infarction is safe and results in a significantly better exercise performance, which might be useful in decreasing the number of equivocal myocardial perfusion studies. PMID- 16047124 TI - Atropine for exercise testing after acute myocardial infarction. PMID- 16047125 TI - Rationale and methods of the integrated biomarker and imaging study (IBIS): combining invasive and non-invasive imaging with biomarkers to detect subclinical atherosclerosis and assess coronary lesion biology. AB - Death or myocardial infarction, the most serious clinical consequences of atherosclerosis, often result from plaque rupture at non-flow limiting lesions. Current diagnostic imaging with coronary angiography only detects large plaques that already impinge on the lumen and cannot accurately identify those that have a propensity to cause unheralded events. Accurate evaluation of the composition or of the biomechanical characteristics of plaques with invasive or non-invasive methods, alone or in conjunction with assessment of circulating biomarkers, could help identify high-risk patients, thus providing the rationale for aggressive treatments in order to reduce future clinical events. The IBIS (Integrated Biomarker and Imaging Study) study is a prospective, single-center, non randomized, observational study conducted in Rotterdam. The aim of the IBIS study is to evaluate both invasive (quantitative coronary angiography, intravascular ultrasound (IVUS) and palpography) and non-invasive (multislice spiral computed tomography) imaging techniques to characterize non-flow limiting coronary lesions. In addition, multiple classical and novel biomarkers will be measured and their levels correlated with the results of the different imaging techniques. A minimum of 85 patients up to a maximum of 120 patients will be included. This paper describes the study protocol and methodological solutions that have been devised for the purpose of comparisons among several imaging modalities. It outlines the analyses that will be performed to compare invasive and non-invasive imaging techniques in conjunction with multiple biomarkers to characterize non flow limiting subclinical coronary lesions. PMID- 16047126 TI - Atherosclerosis imaging and circulating blood biomarkers: insights into subclinical atherosclerotic plaque burden and disease activity. PMID- 16047127 TI - Assessment of coronary artery bypass graft patency by multidetector computed tomography and electron-beam tomography. AB - This case report describes the use of retrospectively ECG-gated 16-slice multidetector computed tomography (MDCT) and electron-beam tomography (EBT) for assessing bypass graft patency in two patients with recurrent angina after coronary artery bypass graft surgery. The results of each tomographic modality were compared to the findings of traditional coronary angiography. In the first patient MDCT showed occlusion of the left internal mammary artery (LIMA) and saphenous vein graft after the second anastomosis. Coronary angiography confirmed these findings. In the second patient EBT showed patency of the LIMA and saphenous vein graft. After the first anastomosis of the saphenous vein graft, the connected vessel filled poorly. Coronary angiography confirmed both grafts to be patent, and detected an occlusion distal to the first anastomosis. These findings support the evidence that both MDCT and EBT are suitable techniques for establishing bypass graft patency by non-invasive means. PMID- 16047128 TI - Computed tomography assessment of coronary bypass grafts: ready to replace conventional angiography? PMID- 16047129 TI - Anomaly of the left atrium in patients with atrial fibrillation detected by ECG gated multi-slice computed tomography. AB - We illustrate pulmonary vein anatomy in 2 patients undergoing multi-slice computed tomography (MSCT) scanning prior to atrial fibrillation ablation, including anomaly of the pulmonary vein. The anomalous variations need to be investigated before catheter procedure, because of the anomalous variations. Non invasive imaging using MSCT could have important clinical implications and provide the physicians useful information on morphological feature in patients with atrial fibrillation before catheter ablation. PMID- 16047130 TI - Aortic dissection with extension to a patent ductus arteriosus. AB - An 81-year-old woman presented with shortness of breath and fever of 3 days duration. An enhanced CT of the chest revealed a patent ductus arteriosus (PDA) communicating with the true lumen of an aortic arch dissection. Neither the aortic dissection nor the PDA was suspected. The patient had no history of a connective tissue abnormality or other condition which would predispose to dissection other than systemic hypertension. It is likely that propagation of the aortic dissection partially re-opened the ductus arteriosus. PMID- 16047131 TI - Cardiac lipoma in a patient with proven arrhythmogenic right ventricular dysplasia: a case report. A huge intramyocardial lipoma. AB - Arrhythmogenic right ventricular dysplasia (ARVD) and primary cardiac tumours are rare conditions among heart diseases. In this case report we present an 18-year old woman with ventricular tachycardia and intracardiac mass originating from interventricular septum. Clinical and pathological evaluation led to the diagnosis of invasive intramyocardial lipoma originated in an ARVD background. To our knowledge, this is the first case with a huge intramyocardial lipoma developed in a patient with ARVD. PMID- 16047133 TI - [Therapy for unilateral vocal fold palsy]. AB - The main symptom of unilateral vocal fold palsy is hoarseness, which can cause considerable disturbance to the patient depending on its extent and the patient's individual situation. Therapy aims at the restitution of a tuneful and resilient voice, which can be achieved by surgical or conservative means, improving the glottal closure and synchronizing the vocal fold vibrations during phonation. Vocal therapy is a common conservative method that may be supported by psychotherapeutic or physical procedures. In surgical therapy, there is a distinction between techniques of endoscopic augmentation by injecting different materials into the vocal folds and transcutaneous laryngeal framework surgery, i.e., transferring the paralyzed vocal fold to the glottal midline. Particularly apt for injection are biocompatible materials amount and position whose can easily be controlled. However, the inevitable resorption of many materials causes deterioration in voice quality. Furthermore, the change of vocal fold morphology obstructs regular phonatory vibration. On the other hand, medialization thyroplasty leads to permanent voice amelioration without a substantial complication rate when performed by experienced surgeons. PMID- 16047134 TI - [Internists' emergency medicine]. PMID- 16047135 TI - [Current concepts in diagnosis and treatment of tachyarrhythmias]. AB - A diagnostic and therapeutic approach of supraventricular and ventricular tachyarrhythmia is always challenging. Several criteria serve to discriminate correctly between these two types of tachycardia. Cardiac arrhythmias are terminated reliably by intravenous (IV) application of antiarrhythmic drugs: adenosine for supraventricular arrhythmia, amiodarone or ajmaline for ventricular tachycardia. Furthermore, AV-nodal tachycardia, atrioventricular reciprocating tachycardia, and typical atrial flutter is treated curatively by radiofrequency ablation during an electrophysiological study. This interventional therapy is well established in patients suffering from ventricular premature contractions or tachycardia originating in the right or left ventricular outflow tract. Aside treatment with an implantable defibrillator, patients with coronary artery disease highly benefit from adjusted pharmaceutical treatment. PMID- 16047136 TI - [Differential diagnosis of anorexia nervosa and depression. A case report of Addison's disease in a female patient]. AB - Similar symptoms make the differential diagnosis of anorexia nervosa and Addison's disease difficult at times: vomiting, weight loss, hypotension, depressed mood, reduced drive, and increased irritability can occur in both disorders. Psychological factors are then frequently presumed to be causative, so that the diagnosis of Addison's disease is made only years after the onset of the disease. Against this background, our case report presents important differential features, particularly with regard to consultative psychiatry. PMID- 16047137 TI - [Interventional endovascular therapy (stenting) for residual stenosis after lysis of the basilar artery]. AB - Despite the introduction of local thrombolytic therapy, there is still a very high mortality rate in basilar thrombosis. We report three cases of successful intra-arterial thrombolysis of acute basilar artery occlusion and consecutive interventional endovascular therapy. In all cases, the acute thrombosis of the basilar artery probably developed from preexisting atherosclerotic stenoses. The clinical outcomes were excellent. We discuss early consecutive stenting as a possible therapeutic option in cases of residual stenosis after successful intra arterial lysis of basilar artery occlusion. PMID- 16047138 TI - Drug-drug interactions among recently hospitalised patients--frequent but mostly clinically insignificant. AB - OBJECTIVE: Patients use and store considerable amounts of drugs. The aim of the present study was to identify potential drug-drug interactions between drugs used by patients recently discharged from the hospital and, subsequently, to estimate the clinical implications of these interactions. METHODS: Patients were visited within 1 week following their discharge from hospital and interviewed about their drug use. Stored products were inspected. We used a bibliography (Hansten and Horn; Wolters Kluwer Health, St. Louis, Mo., 2004) to identify and classify potential drug-drug interactions. RESULTS: In total, 83 surgical and 117 medical patients (n = 200) were included in the survey, of which 139 (70%) were women. The median age was 75 years (range: 24-100 years). Patients stored 2119 medications at home (median: nine per patient; range: 2-44) and used 1622 medications daily or on demand (median: eight per patient; range: 1-24). With respect to those drugs used daily or on demand, 476 potential interactions were identified (126 patients); none were class 1 (always avoid drug combination) and 25 were class 2 (usually avoid combination; 24 patients). Eleven of the potential class 2 interactions involved over-the-counter products (aspirin and ginkgo biloba). Of the 52 drugs involved in potential class 2 interactions, 50 had been used for more than 1 month. According to the hospital case notes, none of the potential class 2 interactions had actually caused adverse effects. CONCLUSION: Although potential drug-drug interactions are highly prevalent, serious and clinically significant interactions are rare among recently hospitalised patients. PMID- 16047139 TI - Suprasellar paraganglioma: a case report and review of the literature. AB - Paragangliomas arising in the suprasellar region are extremely rare. We report a case of suprasellar paraganglioma in a 47-year-old man who presented with amnesia and impaired visual acuity without any endocrine dysfunction. Magnetic resonance imaging (MRI) showed a large enhancing tumour in the suprasellar area. Following subtotal surgical excision, the diagnosis of paraganglioma was confirmed by pathology. In this case report we describe the MRI pattern of suprasellar paraganglioma and review the literature of this uncommon lesion. PMID- 16047140 TI - Spontaneous regression of residual low-grade cerebellar pilocytic astrocytomas in children. AB - BACKGROUND: Cerebellar low-grade astrocytomas (CLGAs) of childhood are benign tumours and are usually curable by surgical resection alone or combined with adjuvant radiotherapy. OBJECTIVE: To undertake a retrospective study of our children with CLGA to determine the optimum schedule for surveillance imaging following initial surgery. In this report we describe the phenomenon of spontaneous regression of residual tumour and discuss its prognostic significance regarding future imaging. MATERIALS AND METHODS: A retrospective review was conducted of children treated for histologically proven CLGA at Great Ormond Street Hospital from 1988 to 1998. RESULTS: Of 83 children with CLGA identified, 13 (15.7%) had incomplete resections. Two children with large residual tumours associated with persistent symptoms underwent additional treatment. Eleven children were followed by surveillance imaging alone for a mean of 6.83 years (range 2-13.25 years). Spontaneous tumour regression was seen in 5 (45.5%) of the 11 children. There were no differences in age, gender, symptomatology, histological grade or Ki-67 fractions between those with spontaneous tumour regression and those with progression. There was a non-significant trend that larger volume residual tumours progressed. CONCLUSIONS: Residual tumour followed by surveillance imaging may either regress or progress. For children with residual disease we recommend surveillance imaging every 6 months for the first 2 years, every year for years 3, 4 and 5, then every second year if residual tumour is still present 5 years after initial surgery. This would detect not only progressive or recurrent disease, but also spontaneous regression which can occur later than disease progression. PMID- 16047141 TI - Sonographic detection of radiographically occult fractures in paediatric ankle injuries. AB - BACKGROUND: In some paediatric ankle injuries, the clinical picture is suggestive of a fracture despite negative three-view radiographs. OBJECTIVE: To determine the effectiveness of high-resolution US to differentiate radiographically occult fractures from sprains. MATERIALS AND METHODS: During a 12-month period, children aged 5-13 years who had sustained an acute ankle injury suspicious of a fracture on clinical examination, but with negative radiography, were referred for high resolution US. Follow-up radiographs were obtained after 2-3 weeks. RESULTS: A total of 20 children were examined. In 13, US did not reveal a fracture; small fractures were detected in 7. All patients with negative US studies had negative follow-up radiographs. In six patients with positive US the follow-up radiographs demonstrated a periosteal reaction. In one child in whom a fracture line identified by US was in the depth of the lateral malleolus, the follow-up radiograph demonstrated an area of increased bone density. CONCLUSIONS: US is effective for the detection of radiographically silent fractures of the paediatric ankle. It may be used as an adjunct to radiography in clinically suspicious, but radiographically negative ankle injuries. PMID- 16047142 TI - Induction of Th1-type immunity and tumor protection with a prostate-specific antigen DNA vaccine. AB - Prostate specific antigen (PSA) is a serum marker that is widely used in the detection and monitoring of prostate cancer. Though PSA is a self-antigen, T cell responses to PSA epitopes have been detected in healthy men and prostate cancer patients, suggesting it may be used as a target for active immunotherapy of prostate cancer. A PSA DNA vaccine (pPSA) was evaluated in mice and monkeys for its ability to induce antigen-specific immune responses. Mice immunized intradermally with pPSA demonstrated strong PSA-specific humoral and cellular immunity. The anti-PSA immune responses were skewed toward Th1, as shown by high IFNgamma and IL-2 production. The immune response was sufficient to protect mice from challenge with PSA-expressing tumor cells. Tumor protection was durable in the absence of additional vaccination, as demonstrated by protection of vaccinated mice from tumor rechallenge. Furthermore, pPSA vaccination induced PSA specific antibody titers in male cynomolgus monkeys, which express a closely related PSA gene. These results demonstrate that vaccination with pPSA may be able to break tolerance and can induce an immune response that mediates tumor protection. PMID- 16047143 TI - Role of immature myeloid cells in mechanisms of immune evasion in cancer. AB - Tumor affects myelopoiesis by inhibiting the process of differentiation/maturation of antigen-presenting cells from their myeloid precursors and by stimulating an accumulation of immature myeloid cells in cancer patients and tumor-bearing mice. These immature myeloid cells can contribute greatly to tumor progression and promote tumor evasion from immune attack: i) by inhibiting development of adaptive immune responses against tumor in lymphoid organs; ii) by migrating into tumor site and differentiating there into highly immune suppressive tumor-associated macrophages. Immature myeloid cells and tumor associated macrophages utilize different JAK/STAT signaling pathways and different mechanisms to control T cell responses, which include increased production of TGF-beta, reactive oxygen species, peroxynitrites, as well as enhanced L-arginine metabolism. Understanding of precise mechanisms, which tumors use to affect differentiation of APC from myeloid cell precursors and inhibit T cell responses, could help to develop new approaches for cancer therapy and substantially improve efficiency of existing cancer vaccination strategies. PMID- 16047144 TI - Morphological appearance, content of extracellular matrix and vascular density of lung metastases predicts permissiveness to infiltration by adoptively transferred natural killer and T cells. AB - We have recently shown that adoptively transferred, IL-2-activated natural killer (A-NK) cells are able to eliminate well-established B16-F10.P1 melanoma lung metastases. However, some B16-F10.P1 lung metastases were resistant to infiltration by the A-NK cells and also resistant to the A-NK cell treatment. The infiltration-resistant (I-R) B16-F10.P1 metastases had a unique "compact" morphology compared to the "loose" morphology of the infiltration-permissive (I P) metastases. Here, we show that I-P loose tumors and I-R compact tumors are also found in lung metastases of mouse Lewis lung carcinoma (3LL), MCA-102 sarcoma, and MC38 colon carcinoma as well as rat MADB106 mammary carcinoma origin. Furthermore, the infiltration resistance of the compact tumors is not restricted to A-NK cells, since PHA and IL-2 stimulated CD8+ T-cells (T-LAK cells) also infiltrated the compact tumors poorly. Analyses of tumors for extracellular matrix (ECM) components and PECAM-1(+) vasculature, revealed that the I-R lesions are hypovascularized and contain very little laminin, collagen and fibronectin. In contrast, the I-P loose tumors are well-vascularized and they contain high amounts of ECM components. Interestingly, the distribution pattern of ECM components in the I-P loose tumors is almost identical to that of the normal lung tissue, indicating that these tumors develop around the alveolar walls which provide the loose tumors with both a supporting tissue and a rich blood supply. In conclusion, tumor infiltration by activated NK and T cells correlates with the presence of ECM components and PECAM-1(+) vasculature in the malignant tissue. Thus, analysis of the distribution of ECM and vasculature in tumor biopsies may help select patients most likely to benefit from cellular adoptive immunotherapy. PMID- 16047145 TI - Phase I/II trial of intraperitoneal 5-Fluorouracil with and without intravenous vasopressin in non-resectable pancreas cancer. AB - BACKGROUND: Systemic palliative treatment with chemotherapy against advanced pancreas cancer has low effectiveness despite considerable toxicity. AIM: To investigate the safety, toxicity and tumour response of intraperitoneal 5 Fluorouracil (5-FU) with intravenous Leucovorin and to monitor 5-FU pharmacokinetics in plasma during intraperitoneal instillation with and without vasopressin in patients with non-resectable pancreas cancer. PATIENTS/METHODS: Between 1994 and 2003, 68 patients with non-resectable pancreas cancer TNM stage III and IV, were enrolled to receive intraperitoneal5-FU instillation 750-1500 mg/m2 and intravenous Leucovorin 100 mg/m2 for two days every third week. Tumour response, performance status and toxicity were recorded. Seventeen patients were also treated with intravenous vasopressin 0.1 IU/minute for 180 minutes, during intraperitoneal 5-FU instillation. Area under the curve (AUC) and peak concentration (Cmax) of 5-FU in plasma were analysed. RESULTS: The treatment was well tolerated with minor toxicity. One complete response (54.1+ months) and 2 partial responses were observed. Time to progression was 4.4 months (0.8-54.1+), and median survival was 8.0 months (0.8-54.1+). There was a significant reduction of 5-FU Cmax in plasma the second day of treatment if vasopressin was used (3.4+/ 2.5 and 6.1+/-5.4 mumol/l, respectively, p<0.05). 5-FU AUC in plasma was not significantly affected by vasopressin either day of treatment. CONCLUSION: Intraperitoneal 5-FU is a safe treatment with low toxicity to patients with non resectable pancreas cancer. Tumour response was 4.4% and median survival time 8.0 months. Addition of vasopressin did not significantly decrease plasma 5-FU AUC but reduced Cmax on day 2 of treatment. PMID- 16047146 TI - Development and validation of limited sampling models for topotecan lactone pharmacokinetic studies in children. AB - PURPOSE: To develop and validate a pharmacokinetic limited sampling model (LSM) for intravenous and oral topotecan pharmacokinetic studies in children. METHODS: Topotecan lactone concentration-time data from five trials were used to develop and validate LSM for intravenous and oral topotecan. Based on full sampling from one intravenous study (30 patients; 195 studies), a LSM for intravenous topotecan was determined using a modification of the D-optimality algorithm. For oral topotecan we used full sampling data from one oral topotecan study (27 patients; 47 studies) to develop an LSM. Accuracy and bias of each LSM were determined relative to the full sampling method. Predictive performance of the LSM was validated using additional data and Monte-Carlo simulations based on these data. RESULTS: LSM for intravenous topotecan includes: 5 min, 1.5, and 2.5 h after the end of the 30 min infusion. The median accuracy (absolute predicted error) and bias (predicted error) are < or =8% and < or =6.1%, respectively. For oral topotecan, the optimal LSM includes: 15 min, 1.5, and 6 h. The median accuracy and bias are 6% and 4%, respectively. CONCLUSIONS: Our results indicate that the optimal sampling times for the intravenous LSM for topotecan in children consist of: predose, and 5 min, 1.5, and 2.5 h after the end of infusion. For oral topotecan the sample times are predose, 15 min, 1.5, and 6 h after dose administration. These LSM are invaluable to children receiving topotecan because it minimizes inconvenience and blood collection. PMID- 16047147 TI - Accelerator mass spectrometry allows for cellular quantification of doxorubicin at femtomolar concentrations. AB - Accelerator mass spectrometry (AMS) is a highly sensitive analytical methodology used to quantify the content of radioisotopes, such as (14)C, in a sample. The primary goals of this work were to demonstrate the utility of AMS in determining total cellular [(14)C]anthracycline concentrations following administration of doxorubicin (DOX) and to develop a sensitive assay that is superior to high performance liquid chromatography (HPLC) for the quantification of [(14)C]anthracycline at the tumor level. In order to validate the sensitivity of AMS versus HPLC with fluorescence detection, we performed three studies comparing the cellular accumulation of DOX: one in vitro cell line study, and two in vivo xenograft mouse studies. Using AMS, we quantified cellular [(14)C]anthracycline content up to 4 h following in vitro exposure at concentrations ranging from 0.2 pg/ml (345 fM) to 2 microg/ml (3.45 microM) [(14)C]DOX. The results of this study show that, compared to standard fluorescence-based HPLC, the AMS method was over five orders of magnitude more sensitive. Two in vivo studies compared the sensitivity of AMS to HPLC using a nude mouse xenograft model in which breast cancer cells were implanted subcutaneously. After sufficiently large tumors formed, [(14)C]DOX was administered intravenously at two dose levels. Additionally, we tested the AMS method in a nude mouse xenograft model of multidrug resistance (MDR) in which each mouse was implanted with both wild type and MDR+ cells on opposite flanks. The results of the second and third studies showed that [(14)C]anthracycline concentrations were significantly higher in the wild type tumors compared to the MDR+ tumors, consistent with the MDR model. Although this method does not discriminate between parent drug and metabolites, the extreme sensitivity of AMS should facilitate similar studies in humans to establish target site drug delivery and to potentially determine the optimal treatment dose and regimen. PMID- 16047148 TI - [Myoepithelial neoplasms of skin and soft tissues]. AB - Myoepithelial neoplasms of skin and soft tissues comprise cutaneous mixed tumor (chondroid syringoma), mixed tumor of subcutaneous and deep soft tissues, myoepithelioma and rare malignant myoepithelioma (myoepithelial carcinoma). Myoepithelial tumors of skin and soft tissues are characterized by an extreme clinicopathological heterogeneity as in other anatomic locations. The neoplasms arise in childhood as well as in adults and are composed of epithelioid, histiocytoid, spindled, plasmocytoid and/or clear tumour cells in varying combinations, and are set in a myxoid or hyalinised intercellular matrix. Immunohistochemically, neoplastic cells stain positively for epithelial markers (pancytokeratin and/or epithelial membrane antigen), and often for S 100 protein. More rarely muscle actin, glial fibrillary acid protein, calponin, and p63 are expressed, whereas desmin is usually negative. The presence of at least moderate cytological atypia is associated with a significant risk for aggressive behavior and propensity for metastasis. PMID- 16047149 TI - Variants of the superior labrum and labro-bicipital complex: a comparative study of shoulder specimens using MR arthrography, multi-slice CT arthrography and anatomical dissection. AB - The purpose of the present study was to evaluate the anatomical variability of the superior labrum and to compare the value of MR arthrography and multi-slice CT arthrography in the diagnosis of variants of the labro-bicipital complex. Forty-three human shoulder specimens (age range and mean age at death, 61-89 years and 78.3 years) were examined with the use of MR arthrography and multi slice CT arthrography prior to joint exploration and macroscopic inspection of the superior labrum and labro-bicipital complex. Two radiologists evaluated MR and CT arthrograms, and the results were compared with macroscopic assessments. Anatomical dissection of all shoulder specimens revealed a sublabral recess in 32/43 (74%) cases. The attachment of the superior labrum was categorised as type 1 in ten (23%) cases, as type 2 in eight (19%), as type 3 in ten (23%), and as type 4 in 14 (33%) cases. One superior labrum showed detachment consistent with a superior labral anteroposterior (SLAP) type 3 lesion. On MR arthrography and CT arthrography the attachment of the superior labrum was categorised in concordance with macroscopic assessments in 79% and 84% of cases, respectively. The anteroposterior extension of sublabral recesses was accurately determined with MR and CT arthrography in 59% and 81% of cases, respectively. The attachment of the superior labrum shows considerable variability. Thus, exact depiction of variants is essential in order to avoid the false positive diagnosis of a superior labral tear (SLAP or Andrews lesion). Both, MR arthrography and multi-slice CT arthrography were effective in the detection and classification of sublabral recesses. PMID- 16047150 TI - [Adjuvant topical interferon-alpha-2b treatment in epithelial tumors of the ocular surface]. AB - PURPOSE: The aim of this study was to evaluate the role of topical interferon alpha-2b in the adjuvant treatment of corneal and conjunctival tumors. METHOD: In this noncomparative, prospective, interventional case series, five patients with histologically proven conjunctival intraepithelial neoplasia (CIN) after primary excision and amniotic membrane transplantation were treated with interferon (IFN) alpha-2b eye drops five times daily over a period of 6 weeks (1 million IU/ml Intron A, Schering). An in situ hybridization technique was used to detect human papillomavirus (HPV) in all specimens. Frequent follow-up was undertaken clinically and photographically for evidence of tumor recurrence. RESULTS: In the follow-up period (13.2+/-4,97 months) no clinical evidence of recurrence with only limited treatment side effects such as mild conjunctival hyperemia were recorded after 6 weeks of interferon. In one CIN specimen HPV 16/18 could be detected. CONCLUSIONS: The combination of excisional biopsy and topical interferon-alpha-2b application seems to be an effective and safe treatment for conjunctival intraepithelial neoplasias. Therefore, we prefer this combined treatment to topical interferon-alpha-2b treatment alone, more destructive approaches such as radiation and cryotherapy, or treatment with antimetabolites such as 5-fluorouracil or mitomycin C. PMID- 16047151 TI - [Fluctuations depending on time of day in measurements of the optic disc using confocal laser canning tomography]. AB - PURPOSE: The aim of this study was to evaluate whether confocal laser scanning tomographic measurements of the optic nerve head depend on the time of day when the examinations are performed. PATIENTS: Thirty-two eyes of 20 subjects repeatedly underwent optic disc confocal laser scanning tomography using the Heidelberg Retina Tomograph I. Operated by a single examiner, measurements were performed at 10 a.m. and 4 p.m. on the same day. RESULTS: Comparing both measurements, differences in optic cup area (0.119+/-0.126 mm(2); p=0.89), neuroretinal rim area (0.120+/-0.131 mm(2); p=0.86), cup volume (0.069+/-0.086 mm(3); p=0.86), rim volume (0.083+/-0.090 mm(3); p=0.51), and retinal nerve fiber layer thickness (0.075+/-0.063 mm; p=0.31) were statistically not significant. Intraocular pressure differences between morning and afternoon ranged between 0 and 14 mmHg. CONCLUSIONS: Confocal laser scanning tomographic measurements of the optic nerve head are not markedly influenced by the time of day. PMID- 16047152 TI - Dorsal premotor areas of nonhuman primate: functional flexibility in time domain. AB - A voluntary motor act requires recognition of the informational content of an instruction. An instruction may contain spatial and temporal information. The recently proved role of the monkey frontal cortex in time computation, as well as in motor preparation and motor learning, suggested that we investigate the relationship between premotor neuron discharges and the temporal feature of the visual instructions. To this purpose, we manipulated the duration of an instructional cue in a visuomotor task while recording unit activity. We found two types of premotor neurons characterised by a discharge varying in relation to the duration of the cue: (1) "motor-linked" neurons, with a specific premotor activity constantly bounded to the motor act; (2) "short-term encoders" neurons, with a premotor activity depending on the cue duration. The cue duration was the critical factor in determining the behaviour of the short-term encoders cells: when the cue ranged from 0.5 s to 1 s, they presented a preparatory activity; when the cue was longer, up to 2 s, they lost their preparatory activity; when the cue was blinked the cells anticipated their discharge. The activity changed in few trials. These data confirm and highlight the role of frontal cortex in encoding specific cues with a temporal flexibility, which may be the expression of temporal learning and represent an extended aspect of cortical plasticity in time domain. PMID- 16047153 TI - The weak bases NH(3) and trimethylamine inhibit the medium and slow afterhyperpolarizations in rat CA1 pyramidal neurons. AB - The weak bases NH(3) and trimethylamine (TMeA), applied externally, are widely used to investigate the effects of increasing intracellular pH (pH(i)) on neuronal function. However, potential effects of the compounds independent from increases in pH(i) are not usually considered. In whole-cell patch-clamp recordings from rat CA1 pyramidal neurons, bath application of 1-40 mM NH(4)Cl or TMeA HCl reduced resting membrane potential and input resistance, inhibited the medium and slow afterhyperpolarizations (AHPs) and their respective underlying currents, mI(ahp) and sI(ahp), and led to the development of depolarizing current evoked burst firing. Examined in the presence of 1 microM TTX and 5 mM TEA with 10 mM Hepes in the recording pipette, NH(3) and TMeA increased pH(i) and the magnitudes of depolarization-evoked intracellular [Ca(2+)] transients, Ca(2+) dependent depolarizing potentials, and inward Ca(2+) currents but reduced the slow AHP and sI(ahp). When internal H(+) buffering power was raised by including 100 mM tricine in the patch pipette, the effects of NH(3) and TMeA to increase pH(i) and augment Ca(2+) influx were attenuated whereas the reductions in the slow AHP and sI(ahp) (as well as membrane potential and input resistance) were maintained. The findings indicate that increases in pH(i) contribute to the increases in Ca(2+) influx observed in the presence of NH(3) and TMeA but not to the reductions in membrane potential, input resistance or the magnitudes of AHPs. The results have implications for the interpretation of data from experiments in which pH(i) is manipulated by the external application of NH(3) or TMeA. PMID- 16047154 TI - Homeopathic treatment of children with attention deficit hyperactivity disorder: a randomised, double blind, placebo controlled crossover trial. AB - An increasing number of parents turn to homeopathy for treatment of their hyperactive child. Two publications, a randomised, partially blinded trial and a clinical observation study, conclude that homeopathy has positive effects in patients with attention deficit hyperactivity disorder (ADHD). The aim of this study was to obtain scientific evidence of the effectiveness of homeopathy in ADHD. A total of 83 children aged 6-16 years, with ADHD diagnosed using the Diagnostic and Statistical Manual of Mental Disorders-IV criteria, were recruited. Prior to the randomised, double blind, placebo controlled crossover study, they were treated with individually prescribed homeopathic medications. 62 patients, who achieved an improvement of 50% in the Conners' Global Index (CGI), participated in the trial. Thirteen patients did not fulfill this eligibility criterion (CGI). The responders were split into two groups and received either verum for 6 weeks followed by placebo for 6 weeks (arm A), or vice-versa (arm B). At the beginning of the trial and after each crossover period, parents reported the CGI and patients underwent neuropsychological testing. The CGI rating was evaluated again at the end of each crossover period and twice in long-term follow up. At entry to the crossover trial, cognitive performance such as visual global perception, impulsivity and divided attention, had improved significantly under open label treatment (P<0.0001). During the crossover trial, CGI parent-ratings were significantly lower under verum (average 1.67 points) than under placebo (P =0.0479). Long-term CGI improvement reached 12 points (63%, P <0.0001). CONCLUSION: The trial suggests scientific evidence of the effectiveness of homeopathy in the treatment of attention deficit hyperactivity disorder, particularly in the areas of behavioural and cognitive functions. PMID- 16047155 TI - Prognostic usefulness of lymphocyte V beta receptor determination in toxic shock syndrome. PMID- 16047156 TI - In vivo allograft rejection in a bony fish Dicentrarchus labrax (L.): characterisation of effector lymphocytes. AB - Lymphoid cell subpopulations involved in allograft rejection in the teleost Dicentrarchus labrax were characterised at the ultrastructural level and quantified by using monoclonal antibodies against T- and B-lymphocytes. T-cells positive for T-cell receptor beta-chain (TcR beta) were detected by reverse transcription/polymerase chain reaction (RT-PCR) and in situ hybridisation by using RNA probes for TcR beta. Flow cytometry detected a similar percentage of T- and B-lymphocytes (around 17%) in the leucocyte-enriched fraction from allografts. Two different types of T-lymphocytes (DLT 15-immunoreactive) infiltrating the allografts were identified by cytomorphology: small cells with high nuclear/cytoplasmic ratio and cells with a higher cytoplasmic content. RT PCR revealed a single band (513 bp) corresponding to the TcR beta. In situ hybridisation showed that TcR beta-positive cells in the grafted muscle fibres were less numerous compared with DLT 15-positive cells, as evidenced in parallel sections, suggesting that cytotoxic cells might express different TcR phenotypes. DLIg 3-immunoreactive Ig-producing lymphocytes had: 1) a high nuclear/cytoplasmic ratio or 2) a larger size similar to that of pre-plasma cells (plasma cells lacked any membrane labelling). PMID- 16047157 TI - Expression and mechanical modulation of matrix metalloproteinase-1 and -2 genes in facial and cranial sutures. AB - Craniofacial sutures create a soft tissue interface between various calvarial and facial bones. Facial and cranial sutures show differences in their surrounding anatomical structures and local mechanical strain environments. Despite previous attempts to identify the expression of matrix metalloproteinase genes (MMPs) in cranial sutures, little is known regarding whether facial and cranial sutures differ in MMP expression. We have investigated the expression of MMP-1 and MMP-2 in the pre-maxillomaxillary suture (PMS; facial suture) and the frontoparietal suture (FPS; cranial suture) in 32-day-old rats with or without the application of cyclic loading. Expression of MMP-1 and MMP-2 was detected by the reverse transcription/polymerase chain reaction technique. At 32 days of postnatal development (n=6), both MMP-1 and MMP-2 were reproducibly expressed in the facial PMS, in comparison with negligible MMP-1 and MMP-2 expression in the cranial FPS. In six age- and sex-matched control rats, cyclic loading at 4 Hz and 1000 mN was applied to the maxilla for two 20-min episodes within a 12-h interval. In some (but not all) cases, cyclic loading induced marked expression of MMP-1 and MMP-2 in the PMS and FPS in comparison with corresponding non-loaded controls. These data confirm our previous finding that short doses of cyclic loading upregulate MMP-2 expression in craniofacial sutures and suggest the possibility that facial and cranial sutures differ in matrix degradation rates during postnatal development. PMID- 16047158 TI - Spermatogonial stem cell transplantation and testicular function. AB - Spermatogonial stem cells (SSCs) are responsible for the continual production of spermatozoa throughout adult life. Interactions between SSCs and the surrounding cells in the seminiferous tubules regulate the biological activity of these cells. Factors involved in the regulation of SSCs are beginning to be defined by animal models and the culture of SSCs in defined media. A critical development in the characterization of SSCs has been the development of the germ cell transplantation technique, which provides the only assay for the presence of SSCs in a population of cells, and which allows the determination of whether SSCs are proliferating or differentiating in culture. This approach has accelerated SSC focused research and promises to provide a better understanding of the factors and mechanisms that regulate these cells. The knowledge provided by this work is also critical to an appraisal of the components of the SSC niche in the seminiferous epithelium. Thus, many aspects of testicular function can be defined by the investigation of SSCs and the factors, cells, and environment that regulate SSCs, thereby leading to a more comprehensive understanding of spermatogenesis. PMID- 16047159 TI - Expression of growth hormone and its receptor in the lungs of embryonic chicks. AB - The lung is well established as being a postnatal target site for growth hormone (GH) action, since pathophysiological states of GH excess and deficiency are both associated with impaired pulmonary function. Pituitary GH is therefore probably involved in normal lung growth or development, although perinatal lung development occurs prior to the differentiation of pituitary somatotrophs and the ontogeny of pituitary GH secretion. The lung itself may, however, be a site of GH production during prenatal development, since a specific GH-response gene (a marker of GH activity) is expressed in the lungs of early chick embryos, in which GH immunoreactivity is widespread in many other peripheral tissues. We have assessed this possibility in embryonic chicks. A 690-bp cDNA, identical in size and nucleotide sequence to the full-length pituitary GH transcript, was amplified by reverse transcription/polymerase chain reaction from total RNA extracted from the lungs of embryos at 11, 13, 15, and 18 days of the 21-day incubation period. This transcript was localized by in situ hybridization to mesenchymal and epithelial cells of the developing lungs, in which specific GH immunoreactivity was similarly located. Intense GH immunoreactivity was also present after embryonic day 15 (ED15) in the smooth muscle surrounding blood vessels in the lung and surrounding the bronchioles. Lung GH immunoreactivity was primarily associated with a 15-kDa protein, rather than the 26-kDa protein in the pituitary gland. After the onset of pituitary GH secretion (at ED17), GH mRNA was barely detectable in the lungs of ED20 embryos, at the start of lung breathing. GH immunoreactivity was, however, still present in some cells in the lungs of ED20 embryos. GH-receptor mRNA and immunoreactivity were also widespread and abundant within the embryonic lung. Lung GH may thus have autocrine or paracrine roles in lung development or in pulmonary function prior to the ontogeny of the pituitary gland and the appearance of GH in peripheral plasma. PMID- 16047160 TI - Tissue- and species-specific expression patterns of class I, III, and IV Adh and Aldh 1 mRNAs in rodent embryos. AB - Alcohol and aldehyde dehydrogenases (ADHs and ALDHs) may be of interest in the pathology of Parkinson's disease (PD) because of their role in protection against toxins and in retinoid metabolism, which is required for growth and development of the mesencephalic dopamine system. In the present study, the spatial and temporal expression patterns of Adh 1, Adh 3, Adh 4, and Aldh 1 mRNAs in embryonic C57BL/6 mice (E 9.5-E19.5) and Sprague-Dawley rats (E12.5-P0) have been investigated by using radioactive oligonucleotide in situ hybridization. High expression of Aldh 1 mRNA was found in the developing mesencephalic dopamine neurons of both mice and rats. Expression of Adh 1 and Adh 4 mRNAs was observed in adrenal cortex and olfactory epithelium in mice. Additionally, Adh 1 was expressed in epidermis, liver, conjunctival, and intestinal epithelium. In rat embryos, expression was less extensive, with Adh 1 mRNA being found in liver and intestines. Adh 3 expression was ubiquitous in both mouse and rat embryos, suggesting a housekeeping function of the gene. Consistent with previous studies in adult rats and mice, our data suggest that Adh 3 is the only ADH class present in rodent brain. Adh and Aldh gene activity in mouse and rat embryos indicate the possible involvement of the respective enzymes in retinoid metabolism and participation in defense against toxic insults, including those that may be involved in the pathogenesis of PD. PMID- 16047161 TI - Immunocytochemical studies on translocation of phosphorylated aquaporin-h2 protein in granular cells of the frog urinary bladder before and after stimulation with vasotocin. AB - We have generated a specific antibody against phosphorylated aquaporin-h2 (pAQP h2) protein to investigate the role of phosphorylation in the translocation of AQP-h2 protein within the granule cells of the urinary bladder of the frog (Hyla japonica). The antibody was generated against a synthetic peptide (ST-160) corresponding to amino acids 255-268, with a phosphorylated Ser-262, a residue that is putatively phosphorylated by protein A kinase. Using this antibody, we found, by Western blot analysis, that phosphorylation of the AQP-h2 protein rapidly increased within 2 min after vasotocin (AVT) stimulation and remained at a higher than normal level for 15 min. Moreover, quantitative immunoelectron microscopy indicated that the location of the AQP-h2 protein dramatically changed after AVT stimulation. Before stimulation, pAQP-h2 protein was localized in only a small number of intracellular vesicles near the nucleus of the granular cells, whereas the labeling density of the intracellular vesicles and the apical membrane rapidly increased after stimulation. This finding was also confirmed by the results of an immunofluorescence study. Thus, phosphorylation of AQP-h2 protein seems to be essential for translocation of the protein from the cytoplasmic pool to the apical plasma membrane of the granular cells in frog urinary bladder. PMID- 16047162 TI - Transdifferentiation of vocal-fold stellate cells and all-trans retinol-induced deactivation. AB - The maculae flavae of the human vocal folds include dense extracellular matrices and compacted cells with a stellate morphology. These vocal-fold stellate cells are thought to participate in the metabolism of extracellular matrices essential in maintaining vocal-fold viscoelasticity required for phonation. We have isolated and cultured these new cells and have tested the hypothesis that they maintain a distinct cellular and biochemical phenotype. We have compared proliferation rates, changes on immunophenotype, and intracellular lipid and vitamin A storage. Vocal-fold stellate cells undergo culture-induced transdifferentiation to a myofibroblast-like phenotype with an altered phenotype resembling, but not identical to, activated hepatic and pancreatic stellate cells. Our results reveal that these cells are capable of responding to exogenous all-trans retinol in culture. Exposure to this synthetic co-factor causes deactivation characterized by decreased proliferation, loss of the activated stellate cell marker, alpha-smooth muscle actin, and restoration of intracellular lipid and vitamin A metabolite storage. These data establish a new and distinct cellular target for future investigations of the viscoelastic properties of the vocal-fold mucosa during normal phonation, aging, vocal-fold scarring, laryngeal fibrosis, and myofibroblastoma. PMID- 16047163 TI - Expression of the noradrenaline transporter and phenylethanolamine N methyltransferase in normal human adrenal gland and phaeochromocytoma. AB - Expression of the noradrenaline transporter (NAT) was examined in normal human adrenal medulla and phaeochromocytoma by using immunohistochemistry and confocal microscopy. The enzymes tyrosine hydroxylase (TH) and phenylethanolamine N methyltransferase (PNMT) were used as catecholamine biosynthetic markers and chromogranin A (CGA) as a marker for secretory granules. Catecholamine content was measured by using high performance liquid chromatography (HPLC). In normal human adrenal medulla (n=5), all chromaffin cells demonstrated strong TH, PNMT and NAT immunoreactivity. NAT was co-localized with PNMT and was located within the cytoplasm with a punctate appearance. Human phaeochromocytomas demonstrated strong TH expression (n=20 samples tested) but variable NAT and PNMT expression (n=24). NAT immunoreactivity ranged from absent (n=3) to weak (n=10) and strong (n=11) and, in some cases, occupied an apparent nuclear location. Unlike the expression seen in normal human adrenal medullary tissue, NAT expression was not consistently co-localized with PNMT. PNMT also showed highly variable expression that was poorly correlated with tumour adrenaline content. Immunoreactivity for CGA was colocalized with NAT within the cytoplasm of normal human chromaffin cells (n=4). This co-localization was not consistent in phaeochromocytoma tumour cells (n=7). The altered pattern of expression for both NAT and PNMT in phaeochromocytoma indicates a significant disruption in the regulation and possibly in the function of these proteins in adrenal medullary tumours. PMID- 16047164 TI - Localization of DJ-1 protein in the murine brain. AB - Mutations in the DJ-1 gene have been identified to cause Parkinson's disease. In humans, nonmutated DJ-1 is expressed in specific brain areas but seems to be expressed by astrocytes rather than by neurons. In contrast, DJ-1 mRNA is mainly found in neurons in the mouse brain. We have investigated the distribution of DJ 1 protein in the mouse brain and found that DJ-1 protein is predominantly expressed by neurons but can also be detected in astrocytes. Consistent with a global role of DJ-1 in the brain, we found immunoreactivity, for example, in cortical areas, hippocampus, basolateral amygdala, the reticular nucleus of the thalamus, zona incerta, and locus coeruleus. Within the substantia nigra, however, DJ-1 is localized in both neuronal and nonneuronal cells, suggesting a distinct role in this area. PMID- 16047165 TI - Olfactory specific chymotrypsin-like serine protease from the aesthetasc tegumental gland of the lobster, Homarus americanus. AB - Numerous proteases and protease inhibitors are expressed in the lobster olfactory organ. One of these proteases, olfactory enriched transcript 03 (OET-03), is particularly interesting because its mRNA is expressed only in one cell type of the olfactory organ of the American lobster, Homarus americanus. We have obtained a full-length cDNA clone of OET-03. The predicted amino acid sequence is equally divided between a novel N-terminal domain and a conserved serine protease catalytic domain at the C-terminus. Heterologous expression in HEK293 cells allowed protease assays demonstrating that OET-03 cleaved a specific serine protease substrate, N-alpha benzoyl-L-arginine p-nitroanilide, but did not cleave a substrate of metalloproteases and cysteine proteases. OET-03 protease activity was significantly inhibited by the chymotrypsin-like protease inhibitor, tosyl-L phenylalanine chloromethyl ketone, but not by the general protease inhibitor, phenylmethylsulfonyl fluoride. Immunoreactivity for OET-03 was detected only in the cells previously shown to contain OET-03 mRNA. The cytoplasm of these cells was filled with enlarged smooth endoplasmic reticulum (a characteristic of secretory cells) that appeared to expand into large electron-translucent areas at the ventral end of the cell. The ventral ends of these secretory cells were apposed to phalloidin-labeled triangular structures reminiscent of the beginnings of the ducts of crustacean tegumental glands. This putative gland was found only in association with the aesthetasc sensory units of the olfactory organ, hence the name, aesthetasc tegumental gland. PMID- 16047166 TI - Developmental changes of Islet-1 and its co-localization with pituitary hormones in the pituitary gland of chick embryo by immunohistochemistry. AB - Although Islet-1 expression in the pituitary gland of early mouse embryo has been previously described, there are no reports concerning the correlation of Islet-1 expression with lineage restrictions in cell types at the later stages of pituitary development. The role of Islet-1 in chickens is also unknown. The purpose of this study was to follow, by using immunohistochemistry, the ontogeny of pituitary Islet-1 and the various cell types that contain Islet-1 throughout chick embryo development. A few Islet-1-immunopositive (Islet-1(+)) cells were first detected in the pituitary primordium in two out of six embryos at embryonic day 5.5 (E5.5), most of the Islet-1(+) cells being ventrally located. As development progressed, many more Islet-1(+) cells were observed throughout the pars distalis. The relative percentage of Islet-1(+) cells amongst the total Rathke's pouch cells was 4.4% at E6.5. This increased significantly, reaching 11.1% by E10.5, followed by no significant change until hatching. Dual immunohistochemistry showed that adrenocorticotrophs, somatotrophs and lactotrophs did not express Islet-1. The cellular types expressing Islet-1 included luteinizing-hormone-positive (LH(+)) gonadotrophs and thyroid stimulating-hormone-positive (TSH(+)) thyrotrophs. The cells co-expressing LH and Islet-1 were initially detected at E6.5, the proportion of LH(+) cells possessing Islet-1 being about 4%; this increased to 63% at E14.5, followed by no significant changes until hatching. TSH and Islet-1 co-localized cells were first observed at E10.5, with about 37% TSH(+) cell expressing Islet-1; this increased to about 50% by E16.5, after which there was no evident change until hatching. These results suggest that Islet-1 is involved in determining the cell lineages, proliferation, differentiation and maintenance of hormone-secreting functions of pituitary gonadotrophs and thyrotrophs of chick embryo. PMID- 16047167 TI - Growth factor impact on articular cartilage subpopulations. AB - We have examined the effects of growth factor stimulation on superficial and growth zone chondrocyte populations. Zonal articular chondrocytes from 8-month old Spanish goat distal femurs were plated in monolayer cultures and stimulated by using insulin-like growth factor I (IGF-I), basic fibroblast growth factor (bFGF), and transforming growth factor-beta1 (TGF-beta1). Gene expression for collagen I and II, aggrecan, and superficial zone protein were evaluated every week for 3 weeks. Finally, proteoglycan and collagen deposition were measured for each experimental group. Major differences existed in the behavior of superficial and growth zone chondrocytes, the most apparent being the higher capacity for protein synthesis by the growth zone population. Variations also existed regarding growth factor treatment. TGF-beta1 had the greatest effect on proliferation over 8 days. With respect to differentiation, IGF-I increased average collagen II gene expression in the growth zone populations in comparison with growth zone controls. IGF-I increased aggrecan gene expression for the same groups. Superficial zone populations exhibited lower collagen II, collagen I, and aggrecan gene expression than the growth zone populations under all conditions. However, superficial zone protein expression was dramatically elevated in superficial zone populations by TGF-beta1. Collagen I expression showed a general increase under all conditions compared with initial values. Combined biosynthesis results showed that the superficial populations secreted little to no collagen, especially collagen II, in comparison with their growth zone counterparts. Glycosaminoglycan production was also much lower than for the growth zone groups. TGF-beta1 and IGF-I increased collagen II production in the growth zone populations. TGF-beta1 increased glycosaminoglycan secretions in the superficial zone populations and in the growth zone populations, whereas IGF-I produced an increase in glycosaminoglycan secretion only in the growth zone populations. Thus, growth factors elicit different proliferation, gene expression, and biosynthesis responses from zonal chondrocyte subpopulations. PMID- 16047168 TI - Application of spectrofluorometry to the prediction of PHB concentrations in a fed-batch process. AB - On-line estimation of biopolymer production during fermentation would be a useful adjunct to the development of strategies for process control and optimization. This study examined the applicability of spectrofluorometry, along with other on line measurements, for the prediction of poly-beta-hydroxybutyric acid (PHB) concentrations in a high-cell density fed-batch fermentation of Ralstonia eutropha. Models previously used for modelling PHB evolution with time are not sufficiently accurate for situations where transient intermediate accumulations or PHB degradation occur. Thus, the mass balance in the model was modified to account for these situations. An estimation algorithm was developed that is based on a hybrid model consisting of a dynamic mass balance of PHB where the main reaction coefficient was regressed with respect to spectrofluorometric data. The regression between the kinetic parameter and the spectrofluorometric data was accomplished using partial least squares (PLS) regression to avoid high sensitivity to noise expected from highly correlated data, such as the spectrofluorometric readings. The model accounts for dynamics of intermediates and in this way allows the prediction of dynamic behaviour in PHB concentrations that cannot be predicted with other reported mathematical models. PMID- 16047169 TI - Application of a mathematical model and Differential Evolution algorithm approach to optimization of bacteriocin production by Lactococcus lactis C7. AB - The effect of pH and temperature on cell growth and bacteriocin production in Lactococcus lactis C7 was investigated in order to optimize the production of bacteriocin. The study showed that the bacteriocin production was growth associated, but declined after reaching the maximum titer. The decrease of bacteriocin was caused by a cell-bound protease. Maximum bacteriocin titer was obtained at pH 5.5 and at 22 degrees C. In order to obtain a global optimized solution for production of bacteriocin, the optimal temperature for bacteriocin production was further studied. Mathematical models were developed for cell growth, substrate consumption, lactic acid production and bacteriocin production. A Differential Evolution algorithm was used both to estimate the model parameters from the experimental data and to compute a temperature profile for maximizing the final bacteriocin titer and bacteriocin productivity. This simulation showed that maximum bacteriocin production was obtained at the optimal temperature profile, starting at 30 degrees C and terminating at 22 degrees C, which was validated by experiment. This temperature profile yielded 20% higher maximum bacteriocin productivity than that obtained at a constant temperature of 22 degrees C, although the total amount of bacteriocin obtained was slightly decreased. PMID- 16047170 TI - [Motor cortical representation in patients with complex regional pain syndrome: a TMS study]. AB - In a group of patients with short- and long-term (chronic) duration of complex regional pain syndrome type I (CRPS I) motor cortical representation was determined, using a transcranial magnetic stimulation (TMS) mapping method. This was done, starting with suprathreshold intensities at the location of the largest MEP amplitude, mapping systematically in all directions. Patients were compared to a group of healthy subjects. In both patient groups we found significantly larger motor cortical representation for the unaffected hand muscles compared to the affected side. This asymmetry was absent in healthy subjects. Such motor cortical representation asymmetry can be considered an effect of altered sensomotor cortical representation. On the other hand, one must also consider the increased use of the unaffected hand and the presence of pain as cortical influencing variables. The real cause must remain speculative at this time. PMID- 16047171 TI - [Stimulation of tiny skin areas for selective stimulation of C fibres]. AB - BACKGROUND: It has been found difficult to stimulate the primary C-fibre afferents separately from those of Adelta fibres. A necessary and sufficient condition for the investigation of the C-fibre system is the selective stimulation of C fibres without activation of Adelta fibres. The stimulation of tiny skin areas allows such a selective activation of C fibres. METHODS AND RESULTS: The main aspects of the method for stimulation of tiny skin areas as well as some results obtained by this method are reported here. The application of this method is compared with applications of other methods that allow an investigation of central processing of human C-fibre input. CONCLUSION: The stimulation of tiny skin areas represents a simple method for selective stimulation of C fibres. PMID- 16047172 TI - Infrared and Raman spectra and vibrational analyses calculated with Moeller Plesset perturbation theory of second order of nitrosoethylene and its chloro derivatives. AB - The conformational and structural stabilities of nitrosoethylene CH2=CH-N=O, chloronitrosoethylene CH2=CCl-N=O, and Dichloronitrosoethylene CCl2=CH-N=O were investigated by ab initio Moeller-Plesset perturbation theory of second order (MP2) calculations using the 6-311+G** basis set to include electron correlation. From the calculations all three were predicted to exist predominantly in the planar trans structure (C=C and N=O bonds are trans to each other) with high trans-cis rotational barriers of about 9 kcal mol(-1) as a result of pronounced conjugation between C=C and N=O bonds. The vibrational frequencies were computed for the three molecules, and also the d1 and d2 deuterated variants for the parent molecule at the MP2 level. Normal coordinate analyses were carried out and the potential energy distributions (PED), among the symmetry coordinates of the normal modes of the molecule were computed. Complete vibrational assignments were made on the basis of normal coordinate analyses for the molecules. The two chlorinated derivatives of nitrosoethylene were also investigated in the same way. As expected, we then find high Raman and infrared intensities in all modes that contain a high content of chlorine movements because vibrations of C-Cl bonds lead to large changes in polarizability, as well as to a large change in dipole moment. However, modes involving double bonds also have quite large intensities. An appreciable number of modes in these molecules are more or less pure symmetry coordinates. PMID- 16047173 TI - Molecular modeling of the intercalation of porphyrins into alpha-zirconium phosphate. AB - In this work, n-alkylamines (number of carbon atoms ranging from 3 to 10) were investigated in detail by molecular modeling as spacers for intercalating porphyrins into alpha-zirconium phosphate (alpha-ZrP). Pre-intercalated n alkylamines can form either a flat monolayer or a canted bilayer in the gallery of alpha-ZrP. Based on the interlayer state and intercalative potential of the two modes in alpha-ZrP, it is suggested that the flat monolayer is a better spacer than the bilayer and that n-propylamine (PA) and n-butylamine (BA) in mobile monolayers are the best spacers among the n-alkylamines studied, as is also found experimentally. The intercalation behavior of TMPyP [5,10,15,20 tetrakis (1-methylpyridinium-4-yl) porphyrin] and several other porphyrins was investigated by calculating the intercalative potential. The calculated results showed that the porphyrins were densely packed in a canted monolayer model, and an increase of polarity of the substituent would facilitate the intercalation of the porphyrins. PMID- 16047174 TI - Nuclear reactivity indices within regional density functional theory. AB - Regional chemical potential values-mu(R) have been obtained with the use of nuclear reactivity indices. Perturbational formulae use values of reactivity indices of isolated molecular fragments. The changes of the parameters (DeltaN(R),{ DeltaQ(i)}i epsilonR) within each fragment determine the value of the regional chemical potential after a chemical reaction. The computational scheme has been tested numerically along the chemical reaction path. We have studied a set of chemical reactions to obtain regional chemical potentials (mu(ts)R) and regional transfer potentials (T(ts)R) for transition states of the following chemical reactions: HF+CO=HFCO, HCl+CO=HClCO, HF+SiO=HFSiO and HF+GeO=HFGeO. The results are reasonable and encouraging. Values of these indices show the possible reactivity directions of the transition states examined. PMID- 16047175 TI - Deterministic center of pressure patterns characterize postural instability in Parkinson's disease. AB - Static posturographic recordings were obtained from six Parkinson's patients and six age-matched, healthy control participants. The availability of vision and visuo-spatial cognitive load were manipulated. Postural sway patterns were analyzed using recurrence quantification analysis (RQA), which revealed differences in center of pressure (COP) dynamics between Parkinson's and control participants. AP COP trajectories for the Parkinson's group were not only significantly more variable than for the control group, but also exhibited distinct patterns of temporal dynamics. The visual manipulation did not differentially affect the two groups. No cognitive load effects were found. The results are generally consistent with the hypothesis that pathological physiological systems exhibit a tendency for less flexible, more deterministic dynamic patterns. PMID- 16047176 TI - The role of dorsal premotor area in reaction task: comparing the "virtual lesion" effect of paired pulse or theta burst transcranial magnetic stimulation. AB - We compared the effect on reaction times of transient interference with function of the dorsal premotor cortex (PMd) using a pair (25-ms interval) of transcranial magnetic stimulation (TMS) pulses with long-term interference produced by a new repetitive TMS paradigm known as "theta burst stimulation" (TBS). Pairs of TMS pulses over left PMd increased choice but not simple reaction times of the right hand if given at the onset of the reaction interval. There was no effect of stimulation over right PMd or at a midline parietal control site (Pz). In contrast, TBS over either left or right PMd increased choice RTs of both hands for at least 5-10 min after the end of TBS. Pairs of TMS pulses over left PMd also increased error rates whereas TBS had no effect on error rates despite the effect on RTs. We suggest that TBS leads to widespread changes in activity and more complex effects on behaviour than expected from the paired pulse TMS and conclude that transient and long-term forms of interference with function may influence behavioural tasks in subtly different ways. PMID- 16047177 TI - Physiological mechanisms of thermoregulation in reptiles: a review. AB - The thermal dependence of biochemical reaction rates means that many animals regulate their body temperature so that fluctuations in body temperature are small compared to environmental temperature fluctuations. Thermoregulation is a complex process that involves sensing of the environment, and subsequent processing of the environmental information. We suggest that the physiological mechanisms that facilitate thermoregulation transcend phylogenetic boundaries. Reptiles are primarily used as model organisms for ecological and evolutionary research and, unlike in mammals, the physiological basis of many aspects in thermoregulation remains obscure. Here, we review recent research on regulation of body temperature, thermoreception, body temperature set-points, and cardiovascular control of heating and cooling in reptiles. The aim of this review is to place physiological thermoregulation of reptiles in a wider phylogenetic context. Future research on reptilian thermoregulation should focus on the pathways that connect peripheral sensing to central processing which will ultimately lead to the thermoregulatory response. PMID- 16047179 TI - Obstructive jaundice in a child with pancreatic hemangioma. AB - We report the case of a 2-year-old boy who presented with obstructive jaundice by pancreatic hemangioma. In this case, a minimal operation for obstructive jaundice without resection of the tumor was performed. Three years after the operation, the pancreatic hemangioma almost disappeared. We discuss treatment of the pancreatic hemangioma in children. PMID- 16047178 TI - Limits to sustained energy intake IX: a review of hypotheses. AB - Several lines of evidence indicate that animals in the wild may be limited in their maximal rates of energy intake by their intrinsic physiology rather than food availability. Understanding the limits to sustained energy intake is important because this defines an envelope within which animals must trade-off competing activities. In the first part of this review, we consider the initial ideas that propelled this area and experimental evidence connected with them. An early conceptual advance in this field was the idea that energy intake could be centrally limited by aspects of the digestive process, or peripherally limited at the sites of energy utilisation. A model system that has been widely employed to explore these ideas is lactation in small rodents. Initial studies in the late 1980s indicated that energy intake might be centrally limited, but work by Hammond and colleagues in the 1990s suggested that it was more likely that the limits were imposed by capacity of the mammary glands, and other works tended to support this view. This consensus, however, was undermined by studies that showed milk production was higher in mice at low temperatures, suggesting that the capacity of the mammary gland is not a limiting factor. In the second part of the review we consider some additional hypotheses that might explain these conflicting data. These include the heat dissipation limits hypothesis, the seasonal investment hypothesis and the saturated neural control hypothesis. Current evidence with respect to these hypotheses is also reviewed. The limited evidence presently available does not unambiguously support any one of them. PMID- 16047180 TI - Necrotizing fasciitis in neonates. AB - Necrotizing fasciitis (NF) is a potentially life-threatening infection of soft tissues. It is characterized by rapid spread of inflammation and infection with widespread necrosis of fascia, subcutaneous tissues, and overlying skin. NF is usually reported in adults with preexisting medical conditions or compromised immune system. It is rare in neonates, and the reported mortality is close to 50% in this population. Less than 70 cases of neonatal NF are reported in literature, most in otherwise healthy neonates and usually attributed to omphalitis, mastitis, or postoperative wound infections. We report our experience of nine neonates who developed NF spontaneously (primary NF) and look at the etiology, clinical presentation, microbiology, management, and outcome. PMID- 16047181 TI - Age-related characteristics of uterine cancer mortality in Japan. AB - BACKGROUND: Several studies have proved that uterine cancer mortality has decreased in Japan over the past 50 years, but the age demographics in this regard are not clear. This study investigated the change pattern of age characteristics for this cancer. METHODS: Cause-specific mortality was identified in the population of Japan and Ireland in the years 1950-2000. Time trends of age specific mortality rate were assessed by fitting linear regression. The change rates between 1950 and 2000 were calculated in comparison with Ireland. The contribution of death from uterine cancer was estimated using cause-elimination life tables. RESULTS: The mortality rate of uterine cancer decreased 59% from 1950 (19.7 per 100,000 women) to 2000 (8.14 per 100,000 women). Age-specific rates for women younger than 70 decreased around 80%, whereas that for women over 85 increased 105% during the same period. Correspondingly, the age of 70 years was the median age for the negative contribution of uterine cancer to life expectancy. Younger than that age, the trend of negative contribution decreased, over that age, it was increased. CONCLUSIONS: Mortality from uterine cancer tended toward the aging in Japan. The general mortality reduction trend was attributed to younger people and adults. These age characteristics can be explained by the increase in life expectancy during the 50-year period. PMID- 16047182 TI - Prospective randomized, double-blinded, placebo-controlled trial of preoperative rofecoxib for pain relief in uterine curettage. AB - OBJECTIVE: To evaluate the analgesic efficacy of preoperative rofecoxib in patients who underwent uterine curettage. MATERIALS AND METHODS: This double blinded, randomized, placebo-controlled trial included 80 women who underwent uterine curettage. Forty women were randomly assigned to rofecoxib 50 mg and 40 women to the placebo. The main outcome measure was the intensity of pain measured by the visual analog scale and categorical pain scores during and after the procedure. Chi-squared, Fisher exact, Student t test, and Mann-Whitney U tests were used for statistical analysis. RESULTS: The intensity of pain was not found to be different between groups over the course of procedure (P>0.05). There were no serious adverse effects in this study. CONCLUSION: The preoperative administration of rofecoxib was not effective in reducing pain in uterine curettage. PMID- 16047183 TI - Central scotoma associated with intraocular silicone oil tamponade develops before oil removal. AB - BACKGROUND: Unexplained sudden visual loss after removal of silicone oil from the eye has recently been described. We report the occurrence and features of unexplained central scotoma developing with silicone oil in situ in the vitreous cavity. METHODS: A retrospective case series of five patients (from two centres) who reported a central scotoma commencing during silicone oil tamponade was studied. All patients had vitrectomy for macula-on retinal detachment, with ultra purified silicone oil tamponade (four out of five had giant retinal tear). Investigations included visual acuity, intraocular pressure, optical coherence tomography, fluorescein angiography, visual fields and electrophysiology. RESULTS: All patients reported a central scotoma that appeared during oil tamponade. Visual acuity fell by a mean of 0.93 LogMAR units after onset of the scotoma. After cataract extraction and oil removal, vision remained reduced by a mean of 0.8 units. The mean duration of oil in the eye was 2.7 months when the scotoma was noted by the patient. Investigations were performed after removal of oil. Fluorescein angiography (FFA) was performed in two cases and optical coherence tomography (OCT) in five patients. No abnormality was demonstrated. Electrophysiology was performed in five patients with pattern electroretinography suggestive of macular dysfunction in four patients. CONCLUSION: This is the first case series describing central scotoma associated with silicone oil in situ. Electrophysiology indicated macular dysfunction in most cases. We suggest that early removal of oil in cases with good visual potential should be considered to avoid this sight-threatening complication. PMID- 16047184 TI - Immunomodulation by topical particle-mediated administration of cytokine plasmid DNA suppresses herpetic stromal keratitis without impairment of antiviral defense. AB - BACKGROUND: We investigated whether the course of herpetic stromal keratitis (HSK) in BALB/c mice could be altered by topical gene-gun-mediated administration of interleukin (IL)-4 or IL-10 plasmid DNA. METHODS: Corneas of BALB/c mice were transfected with plasmids expressing beta-galactosidase (beta-gal), IL-4, IL-10, granulocyte-macrophage colony-stimulating factor (GM-CSF), and pCR3.1 (control) 2 days before Herpes simplex virus-1 (HSV-1; KOS) infection. Development of keratitis and cell infiltration were studied. HSV-1 replication was monitored by plaque assay. Expression of cytokines was detected by enzyme-linked immunosorbent assay. HSV-specific proliferation in the regional lymph nodes and spleens was measured. HSV-1 neutralizing antibody titers and IgG2A/IgG1 ratios were determined. RESULTS: Expression of beta-gal was found in the treated corneas, but not in other tissues. IL-4 or IL-10 plasmid administration induced cytokine production in the corneas. After treatment with 300 psi, the severity of HSK was attenuated (each P<0.05), and the numbers of infiltrating inflammatory cells were lower than in the pCR3.1-treated controls (P<0.001). IL-6, but not IL-1alpha, expression in the cornea was reduced after treatment with IL-4 or IL-10 plasmid DNA. The HSV-1-specific DTH response, corneal Th1 cytokine profile, IgG/IgG2a/IgG1 ratio, neutralizing antibody titers, and virus clearance did not differ between the groups. CONCLUSIONS: Thus, topically administered IL-4 and IL 10 plasmid DNA can lead to a milder course of HSK without impeding viral clearance. The gene gun technique for corneal delivery of plasmid cytokine DNA may be useful for modulating local immune responses without affecting antiviral defense. PMID- 16047185 TI - Differences in reading performance of patients with Drusen maculopathy and subretinal fibrosis after CNV. AB - PURPOSE: To evaluate differences in reading performance and contrast sensitivity on patients with drusen maculopathy and subretinal fibrosis after CNV (choroidal neovascularisation). METHODS: 136 patients (60 with drusen (D), 76 with fibrosis (F)) were studied. Patients were classified according to type of maculopathy and best-corrected visual acuity into groups D1 and F1 (LogMAR 0.2-0.4), groups D2 and F2 (LogMAR>0.4-0.7), and group F3 (LogMAR>0.7-1.3). Reading acuity (in LogRAD) and speed were examined with the Radner Reading charts and compared to the reading speed measured with the long paragraphs of the Zuercher Reading Test. Contrast sensitivity was measured with Pelli-Robson charts. RESULTS: The patients' distance visual acuity was comparable between the drusen and fibrotic CNV groups (D1 versus F1, D2 versus F2). The reading acuity of the corresponding groups D1 and F1 was also comparable, but F2 patients showed a statistically lower reading acuity than D2 patients (P=0.03). All reading speed measurements of the groups F1 and F2 were significantly worse than those of the corresponding groups D1 and D2 (P-values: 0.0005-0.02). The correlation of reading speed between the Radner and Zuercher texts was very high (r=0.73-0.94). The contrast sensitivity was significantly lower in all groups compared with group D1 (P<0.001), but comparable for groups F1, F2, and F3. CONCLUSIONS: Despite comparable results in distance visual acuity, patients with subretinal fibrosis after CNV had a greater reduction in reading ability than the patients with drusen. The distance visual acuity measurements alone, therefore, underestimate the real-life conditions and impact of AMD. PMID- 16047186 TI - Assessment of room temperature influence on finger blood flow response induced by short-term grasping of vibrating handle. AB - OBJECTIVE: To investigate the influence of room temperature on finger blood flow (FBF) change in healthy subjects exposed to short-term grasping of a vibrating handle under different room temperatures. METHODS: FBF was measured using a blood flowmeter in six male subjects on the dorsum of the middle phalanx of third finger in both hands once at the end of every minute for an equal duration of 5 min at pre-exposure, during exposure to grasping of vibrating handle with sinusoidal vibration and after exposure. Vibration was generated with a frequency of 125 Hz and an rms acceleration of 40 m/s(2). Measurements were conducted in four room temperatures of 15+/-1, 20+/-1, 25+/-1 and 30+/-1 degrees C. RESULTS: Compared with the baseline measurements in the exposed hand during grasping of vibrating handle most significant increase in FBF was observed at 15+/-1 degrees C (P<0.001) and least at 30+/-1 degrees C (P<0.05), and after vibration least significant FBF was found at 25+/-1 degrees C (P<0.05). In case of the unexposed hand significant increase in FBF was exhibited at 20+/-1 degrees C (P<0.01) and 30+/-1 degrees C (P<0.01) during vibration, and only at 15+/-1 degrees C (P<0.05) after vibration. CONCLUSIONS: Response in FBF due to grasping of vibrating handle was of different patterns from the baseline measurement under different room temperature conditions in both exposed and unexposed hands and it was influenced by room temperature. Overall, the influence was greater at lower test room temperature, inducing more significant increase in FBF. PMID- 16047187 TI - Immunolocalization and distribution patterns of type IV collagen alpha chains in oral mucosal melanoma. AB - The basement membrane (BM) is mainly composed of type IV collagen, which is composed of triple combinations of six distinct alpha (alpha) chains in a tissue specific manner. The six collagen chain-specific antibodies (alpha1-alpha6) were used to examine the BMs of the oral epithelium (OE) and tumor clusters in oral mucosal melanoma (OMM). Eight OMM cases were examined. Results showed that the alpha1 and alpha2 chains were constantly detected at the BM of the normal OE as well as at the OE with atypical melanocytic proliferation and in invasive melanoma with nodular nests. The alpha1 and alpha2 chains were intermittently detected in in situ OMM, early invasive OMM and advanced invasive OMM with sheet like nests. Gradual loss of alpha5 and alpha6 from the OE with atypical melanocyte through in situ OMM and early invasive OMM was observed. These findings suggest that changes in the immunolocalization and distribution patterns of type IV collagen alpha chains are associated with the progression of OMM. The distribution pattern of type IV collagen alpha chain varies depending on the architecture of the nest. PMID- 16047188 TI - Expression of chemokine receptor CXCR4 during chick embryo development. AB - The chemokine receptor CXCR4 plays a decisive role in physiological cell migration both in developmental processes and adult tissues; it has also been implicated in metastasis formation of different human cancers (Balkwill 2004) and in HIV pathogenesis (Murdoch 2000). Here we present the expression pattern of this important chemokine receptor CXCR4 in the chick embryo. A dynamic expression pattern can be detected beginning as early as the gastrulation stages until the observed stage of HH28. During gastrulation, expression was observed in the epiblast at the level of the primitive streak and in the endoderm. Later, expression was noticeable in the ventral foregut portal, developing somites, tail bud, neural tube, the intermediate mesoderm, Wolffian duct, the lateral plate mesoderm and the developing blood vessels. Our descriptive data suggest a role for CXCR4 in gastrulation and other morphogenetic events connected with angiogenesis and kidney development. PMID- 16047189 TI - Detection of disseminated lung cancer cells in regional lymph nodes by assay of CK19 reverse transcriptase polymerase chain reaction and its clinical significance. AB - PURPOSE: To set up a molecular method (i.e. RT-PCR) that can be used to detect disseminated tumor cells (DTCs) in regional lymph nodes (LNs) in patients with lung cancer and to evaluate its clinical significance. METHODS: Cytokeratin 19 (CK(19)) was used as marker. Serial dilution study for LC-5 cells (a lung squamous cell line) was performed to detect sensitivity of the molecular protocol. Regional LNs (n = 261) and primary lung cancer tissue (n = 40) were obtained from 40 patients with lung cancer who underwent lobectomy or pneumonectomy. They were randomly categorized into two groups: group I (LN-based study, n = 20) and group II (patient-based study, n = 20). Each LN was halved. One half of a LN was subjected to histological examination (HE) and the other half was subjected to RT-PCR amplification of CK(19) mRNA. The effect on survival was analyzed. The cumulative survival was calculated by the Kaplan-Meier method and compared by the log rank test. The Cox model analyzed the prognostic factors. RESULTS: CK(19) mRNA expressed in all tumor tissues as well as LC-5, PAa cells (a lung adenocarcinoma cell line), but not in normal control LNs. Serial dilution study for LC-5 cells demonstrated that CK(19) mRNA was detectable at a concentration as low as 10 LC-5 cells in 1x10(7) LN cells. There was no significant difference between the detecting result of single LN and that of mixed LNs (P > 0.05). In 18 of 40 patients, the metastasis in regional LNs was found by both HE and RT-PCR. Of 22 patients without pathologically involved nodes, six (27%) were found to express CK(19) mRNA in regional LNs. According to the results of regional LNs in 40 patients by molecular assay, the presence of the CK(19) product in LNs was related to tumor size (chi(2) = 5.76, P < 0.025) as well as cell differentiation of the tumor (chi(2) = 7.08, P < 0.01). Following a median observation time of 26 months (range, 4 to 60 months), patients with DTCs in nodes showed significant shorter disease-free survival duration than node negative patients (log-rank test, P = 0.001). The independence of this prognostic significance was demonstrated by a multivariate analysis (Cox regression model, P = 0.004). The results diagnosed by HE had no significant effect on prognoses (P = 0.455). CONCLUSIONS: Comparing with HE, RT-PCR can make more accurate assessment of metastatic status in LNs, which is helpful for screening the patients in whom the early subclinical metastasis exists and disclosing the intrinsic regulation of malignant metastasis. The presence of DTCs in LNs is an independent factor for prognosis. Molecular detection of DTCs in LNs is a supplement for current tumor staging in lung carcinoma. PMID- 16047191 TI - Distribution of gadomelitol in a human breast tumor model in mice. AB - The study evaluates the tumor distribution of the rapid clearance blood pool agent (RCBPA) gadomelitol, in a breast tumor model. Different techniques were used : (1) tissue gadolinium concentrations measured by inductively coupled plasma atomic emission spectroscopy (ICP-AES), (2) whole body quantitative autoradiography using radiolabeled [153Gd] gadomelitol and (3) dynamic contrast enhanced MRI with compartmental analysis. An accumulation of gadomelitol in tumors compared to muscle was observed 30 min and 3 h post injection (p.i.). Thirty minutes p.i., the gadomelitol tumor distribution evaluated by autoradiography showed a marked difference between the rim and the center, whereas both areas showed comparable concentrations after 3 h. Using dynamic contrast-enhanced MRI, three phases could be observed during the 1 hour observation period: (1) rapid tumor uptake within the first few minutes post injection (2) a progressive increase in tumor signal enhancement over 10 min and (3) a steady-state phase. Average +/- SD (n=5) transendothelial permeability K(PS) and the fractional blood volume fBV were 12.2+/-1.6 microl/min(-1)/g and 5.4+/-0.2% respectively. Due to its slow extravasation and high tumor residence time, gadomelitol may potentially be useful to improve characterization between benign versus malignant tumors using dynamic MRI. PMID- 16047190 TI - Dynamic analysis of metabolic effects of chloroacetaldehyde and cytochalasin B on tumor cells using bioelectronic sensor chips. AB - PURPOSE: To study the interplay of drugs and energy metabolism of tumor cells, metabolic changes induced by chloroacetaldehyde and cytochalasin B were analyzed in colon carcinoma cells LS174T. METHODS: O(2)-consumption and extracellular acidification were recorded using a bioelectronic sensor-chip system, which monitors these parameters in a culture continuously for at least 24 h. In parallel cultures cell number, cellular ATP-content, mitochondrial transmembrane potential, and the content of reactive oxygen species (ROS) were determined. RESULTS: When cell death was induced by chloroacetaldehyde (50 muM), the rate of acidification declined gradually for the next 15 h, while O(2)-consumption decreased rapidly within 30 min. This correlated with a loss in mitochondrial potential. However, cellular ATP-level showed a transient increase at 2 h; also ROS levels increased up to 6 h. In cells treated with cytochalasin B (2 muM), which inhibits glucose uptake, the rate of O(2)-consumption increased and the acidification activity dropped, even upon glutamine depletion. Mitochondrial membrane potential transiently increased after 1 h, while ATP-content decreased; there was no change in the level of ROS. CONCLUSION: The pattern of changes in basic energy metabolism differs with the type of cell death and growth inhibition involved in the cytotoxic action of two different drugs. PMID- 16047192 TI - Development of new rat monoclonal antibodies with different selectivities and sensitivities for 2,4,6-trinitrotoluene (TNT) and other nitroaromatic compounds. AB - Five new rat monoclonal antibodies (mAbs) for 2,4,6-trinitrotoluene (TNT) and other nitroaromatic compounds, including, especially, the metabolite 2-amino-4,6 dinitrotoluene (2-ADNT), are described. Five heterogeneous, competitive enzyme linked immunosorbent assays (ELISAs) were developed. Assay 1 uses mAb DNT4 3F6 as recognition element and gives a standard curve for TNT in 40 mmol L(-1) phosphate buffered saline (PBS) with a test midpoint (IC50) of 0.26+/-0.08 microg L(-1) (n=20). Assay 2 (mAb DNT4 4G4) has an IC50 of 0.35+/-0.07 microg L(-1) (n=18), assay 3 (mAb DNT4 1A3) has an IC50 of 0.73+/-0.14 microg L(-1) (n=15), and assay 4 (mAb DNT4 1A7) has an IC50 of 2.32+/-0.70 microg L(-1) (n=15). Assay 5 (mAb DNT2 4B4) is very selective for 2-ADNT and has an IC50 of 8.5+/-1.7 microg L(-1) (n=15) in PBS. These antibodies for nitroaromatic compounds differ not only in their sensitivity but also in their selectivity. Major cross-reactants are 1,3,5 trinitrobenzene, 2-ADNT, 4-amino-2,6-dinitrotoluene (4-ADNT), 2,4-dinitroaniline, 3,5-dinitroaniline, and 2,6-dinitroaniline. Although assay 5 is not highly sensitive, the mAb DNT2 4B4 in this assay is highly selective for 2-ADNT. Of all the compounds tested, only 2,4-dinitroaniline and 3,5-dinitroaniline had relevant cross reactivities, 18% and about 26%, respectively. Two ELISAs, using mAbs DNT4 3F6 and DNT2 4B4, were used to analyze different concentrations of TNT and 2 ADNT, respectively, in three different surface water matrices (river and lake water). Both assays were affected by the matrix, but usually performed well (recovery within the range 70-120%). In addition, these ELISAs were used to analyze mixtures of TNT, 2-ADNT, and 4-ADNT, at three different concentrations, in the same water matrices. A different recognition pattern was clearly visible with both assays and depended on the cross reactivities of the corresponding mAb. PMID- 16047193 TI - Characterization of nicotine's ability to serve as a negative feature in a Pavlovian appetitive conditioning task in rats. AB - RATIONALE: Pavlovian feature negative discriminations have been widely used to understand inhibitory conditioning processes using exteroceptive stimuli. Comparatively little is known about inhibitory conditioning processes using a drug state as a negative feature. A negative feature signals that presentation of a conditional stimulus (CS) will not be paired with an unconditioned stimulus. OBJECTIVES: The present research examined whether nicotine served as a negative feature and started characterizing its properties. METHODS AND RESULTS: In acquisition, rats received intermixed saline and nicotine (0.4 mg/kg, base) sessions. On saline sessions, a 15-s light CS was paired with 4-s access to sucrose; the CS was presented on nicotine sessions, but sucrose was withheld. The discrimination was acquired with more goal tracking during the CS on saline sessions. Nicotine's inhibition of this conditioned response (CR) was sensitive to nicotine dose (ED50=0.225) and injection to testing interval (CR returned at 200 min). Mecamylamine pretreatment, but not hexamethonium, produced a loss of inhibitory control by nicotine suggesting a role for central nicotinic acetylcholine receptors. Amphetamine, bupropion, arecoline, and chlordiazepoxide, but not caffeine, substituted for the nicotine feature. However, in locomotor tests, amphetamine and bupropion increased activity; arecoline and chlordiazepoxide decreased activity. For this reason, the motor effects of these ligands could not be dissociated from substitution via shared stimulus properties. CONCLUSIONS: This feature negative task provides a preclinical model for studying how drug states inhibit responding, although identifying the process(es) mediating CR inhibition will require further research. PMID- 16047194 TI - A selective test for antidepressant treatments using rats bred for stress-induced reduction of motor activity in the swim test. AB - RATIONALE AND OBJECTIVE: This paper describes a new procedure for detecting effective antidepressant treatments. The procedure uses the swim-test susceptible (Susceptible) rat which has been selectively bred to show decreased struggling behavior in a swim test after exposure to a mild stressor. The ability of treatments to block this decrease in swim-test activity was assessed as a method for detecting effective antidepressants. RESULTS: In both male and female Susceptible rats, chronic (14-day) treatment with different antidepressant drugs delivered via osmotic minipump [i.e., three tricyclics (desmethylimipramine, imipramine, amitriptyline), two selective serotonin reuptake inhibitors (fluoxetine and sertraline), a monoamine oxidase inhibitor (phenelzine), and two atypical antidepressants (venlafaxine and bupropion)] all prevented the stress induced decrease in swim-test struggling normally shown by these rats. Electroconvulsive shock had a similar effect. Unlike antidepressant drugs, 14-day treatment with various non-antidepressant drugs [i.e., a stimulant (amphetamine), an anxiolytic (chlordiazepoxide), an antihistamine (chlorpheniramine), and an anticholinergic (scopolamine)] did not have this effect. Antidepressant drug treatment for 1 day (i.e., acute treatment) was also ineffective in this test. The procedure described above requires use of the Susceptible rat--swim test resistant rats (i.e., rats selectively bred to be resistant to decreased swim test activity after exposure to stressful conditions) showed no significant differences in swim-test behavior between stress and nonstress conditions after 14-day drug treatment, and randomly bred Sprague-Dawley rats did not show a decrease in swim-test activity following exposure to the mild stressor that is the basis for the test. CONCLUSION: These results suggest that the procedure described here, which uses a rat subject that has been bred for vulnerability to stressful conditions, may be a selective screening technique for effective antidepressant treatments. PMID- 16047195 TI - Attentional and motivational deficits in rats withdrawn from intravenous self administration of cocaine or heroin. AB - RATIONALE: Identifying the long-term neurocognitive sequelae of drug addiction may have important implications for understanding the compulsive, chronically relapsing nature of this brain disorder. OBJECTIVES: Our aim was to investigate the consequences of chronic intravenous self-administration of cocaine or heroin on visual attentional processes in rats. METHODS: Adult male rats were pretrained on a five-choice serial reaction time task (5-CSRTT) of sustained visual attention and impulsivity and later trained to self-administer cocaine or heroin intravenously during multiple 'long-access' self-administration cycles. Control rats had identical training and surgical experience, but received passive infusions of saline during self-administration sessions. Executive cognitive processes of selection and inhibitory response control were evaluated 24 h after drug discontinuation and for a further 6 days prior to the next cycle of self administration. RESULTS: Findings indicate similar behavioural disturbances on the five-choice task in cocaine- and heroin-withdrawn rats with significantly impaired attentional accuracy, increased omissions and slower latencies to respond correctly during the early, but not late, withdrawal period. The self administration of either drug was not associated with significant alterations in impulsive actions, and there was no evidence of persistent alterations in visual attentional performance. However, unlike rats self-administering cocaine, the motivation to collect food reward on the 5-CSRTT was significantly reduced in heroin-withdrawn animals for a period of at least 6 weeks. CONCLUSIONS: These data, together with recent findings of attentional dysfunction during the withdrawal of intravenous self-administration of amphetamine, suggest that generically different drugs of abuse produce similar disturbances in visual attentional performance during the early withdrawal period. PMID- 16047196 TI - Estimating the prevalence and strength of non-independent predator effects. AB - Understanding whether multiple predator species have independent effects on shared prey is critical for understanding community dynamics. We describe the prevalence and strength of non-independence between predators by quantifying the prey's risk of predation and the degree to which it deviates from the risk predicted from a null model of independent predator effects. Specifically, we document how frequently non-independent effects occur among ten different multiple predator combinations with mayfly larvae as prey. These predator combinations vary both predator density and predator species richness. Overall, the predator effects were non-independent and translated to an average of 27% fewer prey being consumed compared to independent predator effects. Non independence of this magnitude is likely to have population level consequences for the prey and influence the distribution or prey preference of predators. Closer inspection shows that much of the risk reduction in this system is weak, to the point of being indistinguishable from independent predator effects, while few effects are strong. This pattern of many weak interactions and few strong ones parallels the pattern of interaction strengths documented previously in intertidal communities. Consequently, understanding strong interactors in multiple predator systems may help us understand the importance of a species. PMID- 16047197 TI - [When is aftercare useful in renal cell cancer?]. AB - Diagnosis of tumor recurrence and of therapy-related side effects as well as psychological support are the main goals of a surveillance program of cancer patients. While the latter may represent a time-consuming effort, most diagnostic procedures are expensive. Whether we can efficiently detect tumor recurrence in renal cell carcinoma depends on various parameters of the recurrent disease (e.g., frequency, localization, or therapeutic options). Available data lend support to "lean" follow-up strategies in patients with renal cell carcinoma. PMID- 16047198 TI - Enhanced resistance to early blight in transgenic tomato lines expressing heterologous plant defense genes. AB - Genes coding for an iris ribosomal-inactivating protein (I-RIP), a maize beta glucanase (M-GLU), and a Mirabilis jalapa antimicrobial peptide (Mj-AMP1) were separately introduced into tomato (Lycopersicon esculentum cv. Sweet Chelsea) cotyledons via Agrobacterium tumefaciens-mediated transformation. Transgenic lines carrying each of the transgenes were confirmed for integration into the tomato genome using Southern blot hybridization. Transcription of I-RIP, M-GLU, and Mj-AMP1 genes in various transgenic lines was determined using Northern blot analysis. Plants of selected transgenic lines were inoculated with a 2-3x10(4) conidial spores/ml suspension of the fungal pathogen Alternaria solani, the causal agent of tomato early blight. Compared to control (non-transformed) plants, two transgenic lines carrying either a M-GLU or Mj-AMP1 showed enhanced resistance to early blight disease. None of the four lines carrying the I-RIP transgene showed increased resistance to early blight. PMID- 16047199 TI - Cholodny-Went revisited: a role for jasmonate in gravitropism of rice coleoptiles. AB - Gravitropism is explained by the Cholodny-Went hypothesis: the basipetal flow of auxin is diverted laterally. The resulting lateral auxin gradient triggers asymmetric growth. However, the Cholodny-Went hypothesis has been questioned repeatedly because the internal auxin gradient is too small to account for the observed growth asymmetry. Therefore, an additional gradient in indolyl-3-acetic acid (IAA) sensitivity has been suggested (Brauner and Hager in Planta 51:115 147, 1958). We challenged the Cholodny-Went hypothesis for gravitropism of rice coleoptiles (Oryza sativa L.) and found it to be essentially true. However, we observed, additionally, that the two halves of gravitropically stimulated coleoptiles responded differentially to the same amount of exogenous auxin: the auxin response is reduced in the upper flank but normal in the lower flank. This indicates that the auxin-gradient is amplified by a gradient of auxin responsiveness. Hormone contents were measured across the coleoptile by a GC MS/MS technique and a gradient of jasmonate was detected opposing the auxin gradient. Furthermore, the total content of jasmonate increased during the gravitropic response. Jasmonate gradient and increase persist even when the lateral IAA gradient is inhibited by 1-N-naphtylphtalamic acid. Flooding with jasmonate delays the onset of gravitropic bending. Moreover, a jasmonate deficient rice mutant bends more slowly and later than the wild type. We discuss a role of jasmonate as modulator of auxin responsiveness in gravitropism. PMID- 16047200 TI - Bacterial luciferase activity and the intracellular redox pool in Escherichia coli. AB - In this study, we analyzed the activity of a bacterial luciferase (LuxAB of Vibrio fischeri) expressed under the control of a consensus-type promoter, lacUV5, in Escherichia coli, and found that activity declines abruptly upon entry into the stationary growth phase. Since this decline was reproducibly observed in strains cultured in various growth media, we refer to this phenomenon as ADLA (Abrupt Decline of Luciferase Activity) and define the time point when activity begins to decline as T (0). Because the levels of luciferase proteins (LuxA and LuxB) remained constant before and after T (0), ADLA cannot be due to the repression of luciferase gene expression. Further analyses suggested that a decline in the supply of intracellular reducing power for luciferase was responsible for ADLA. We also found that ADLA was alleviated or did not occur in several mutants deficient in nucleoid proteins, suggesting that ADLA is a genetically controlled process involved in intracellular redox flow. PMID- 16047202 TI - [Anesthesia during pregnancy]. AB - The need for general or regional anesthesia due to nonobstetric surgery occurs in up to 2% of all pregnancies. Safe perioperative care is a challenge to anesthetists. Profound knowledge of physiological and pathophysiological changes during pregnancy and the possible influence of different drugs and anesthesia techniques on mother and fetus is necessary. Although data from randomized controlled trials is missing, safe anesthesia during pregnancy is well documented for nearly every operative procedure. Interdisciplinary cooperation between surgeon, obstetrician, neonatologist, and anesthetist is the key to optimal patient care. PMID- 16047201 TI - Structural and functional analysis of 5S rRNA in Saccharomyces cerevisiae. AB - 5S rRNA extends from the central protuberance of the large ribosomal subunit, through the A-site finger, and down to the GTPase-associated center. Here, we present a structure-function analysis of seven 5S rRNA alleles which are sufficient for viability in the yeast Saccharomyces cerevisiae when expressed in the absence of wild-type 5S rRNAs, and extend this analysis using a large bank of mutant alleles that show semi-dominant phenotypes in the presence of wild-type 5S rRNA. This analysis supports the hypothesis that 5S rRNA serves to link together several different functional centers of the ribosome. Data are also presented which suggest that in eukaryotic genomes selection has favored the maintenance of multiple alleles of 5S rRNA, and that these may provide cells with a mechanism to post-transcriptionally regulate gene expression. PMID- 16047203 TI - FLAG-IDA in the treatment of refractory/relapsed adult acute lymphoblastic leukemia. AB - Relapsed or refractory adult acute lymphoblastic leukemias (ALL) have poor prognosis. The strategy for treating these patients is through reinduction chemotherapy followed by allogeneic stem cell transplantation, provided that the toxicity of the salvage regimen is acceptable. Twenty three patients with relapsed/refractory adult ALL were treated with fludarabine, cytarabine, granulocyte colony-stimulating factor, and idarubicin (FLAG-IDA). Five patients had primary refractory disease, and 18 were in first relapse. Nine (39.1%) patients achieved complete remission (CR) following salvage therapy, whereas 13 (56.5%) patients were refractory, and one patient died in aplasia due to infection. In patients achieving remission, the median time to reach absolute neutrophil count (ANC) more than 0.5x10(9)/l and 1x10(9)/l was 20 (range 16-25) and 24 (range 20-28) days from the start of chemotherapy, respectively. Platelet levels of more than 20x10(9)/l and 100x10(9)/l were achieved in a median time of 23 (range 19-25) and 33 (range 28-39) days, respectively. Fever more than 38.5 degrees C was observed in 18 of 23 patients (78.2%), 13 had fever of unknown origin, and 5 had documented infections. Nonhematological side effects, consisting mainly of mucositis (18/23 or 78.2%) and transient liver toxicity increase (10/23 or 43.4%), were generally tolerated. All nine patients who achieved CR received a second course with FLAG-IDA, and seven patients underwent allogeneic stem cell transplantation (four from a matched donor, one from a mismatched donor, and two from an unrelated donor), while two did not reach that stage due to early relapse from CR. The median overall survival (OS) for all 23 patients was 4.5 (range 1-38) months; for the nine responders, the disease-free survival (DFS) and the OS were 6 (range 3-38) and 9 (7-38) months, respectively; the seven patients who received allogeneic stem cell transplantation had a DFS of 10 (range 7-38) months. In our experience, FLAG-IDA is a well-tolerated regimen in relapsed/refractory ALL patients; the toxicity is acceptable, enabling patients who have achieved CR to receive allogeneic transplantation. PMID- 16047204 TI - Colony-stimulating factors in the management of neutropenia and its complications. AB - Granulocyte colony-stimulating factor (CSF) and granulocyte-macrophage CSF are potent drugs used to increase neutrophil counts after myelosuppressive chemotherapy. However, in various indications, the use of CSFs has no clinical benefit with regard to morbidity or mortality from infectious complications, frequency of antibiotic use, or rate of hospitalization. Thus, the application of CSFs should be limited to indications with proven clinical benefits or evidence of cost-effectiveness. This review will provide an overview of the state-of-the art use of CSFs in chemotherapy-associated neutropenia, transplantation, and bone marrow failure syndromes. In addition, recently developed drugs for accelerated hematopoietic recovery will be presented. PMID- 16047205 TI - Complement profile in childhood immune thrombocytopenic purpura: a prospective pilot study. AB - Immune thrombocytopenic purpura (ITP) is a frequent, usually acute hematologic complication of viral diseases in children. Its evolution is usually benign with spontaneous resolution. Some authors have raised the hypothesis of a possible role of the complement system in the pathogenesis of ITP. In the present study, we measured the complement system components in 25 children with ITP. The complement system was abnormal in most patients with ITP as compared with the control group: 20 of the 25 patients had at least one low-component level. The most affected components were properdin, factor H (p=0.005 and p=0.001, respectively), C1q, C9, and factor B. Our results may indicate a possible role for the complement system in the pathogenesis of pediatric ITP. PMID- 16047206 TI - Ganglioneuroma : primary tumor or maturation of a suspected neuroblastoma? AB - Ganglioneuroma is a benign neurogenic tumor. These tumors are originating from neuroepithelium along sympathetic ganglia. Main localization is the mediastinum in children older than 10 years. An association with malignant neuroblastoma is rarely observed and it still remains a topic under current discussion. We describe the clinical course of a 17 year-old female patient with a large presacral mass causing amenorrhoea and weight loss. Eleven years before presentation, an incidental urine test showed an elevation of vanillylmandelic acid (VMA) and homovanillic acid (HMA) and a neuroblastoma was suspected. However, further investigations showed no tumor and the test results turned out to be normal within 1 year. Now, a malignant neurogenic tumor was again suspected, but a CT-guided biopsy revealed a benign tumor. The mass was originating from the left sacral nerve roots. A tumor resection via a dorsal approach was performed. Final histology showed a differentiated ganglioneuroma. This is, to our knowledge, the first report describing a patient with elevated VMA/HMA and suspected neuroblastoma who developed a ganglioneuroma 11 years later. The association of ganglioneuroma and neuroblastoma and the abnormal urine tests pointing toward a neuroblastoma 11 years ago remains unclear and the possible answers are discussed in our report. PMID- 16047207 TI - Three-dimensional relation of skin markers to lumbar vertebrae of healthy subjects in different postures measured by open MRI. AB - The debate is to which extent external skin markers represent true underlying vertebral position and motion. Skin markers and lumbar vertebrae L3 and L4 were examined by vertically open magnetic resonance imaging (MRI) within different postures to investigate whether, and to which extent the position and orientation of skin markers represent the corresponding information of assigned underlying vertebra. Nine healthy volunteers sat within an open MRI scanner in five different seating postures: upright, low flexion, heavy flexion, upright left turn and upright right turn. Skin markers were fixed at lumbar levels L3 and L4. A set of landmarks defines corresponding positions on the vertebrae. Translation vectors quantify the change of co-ordinates while changing position. Orientation (Cardan-angles) of each level in space was calculated from co-ordinates of three skin-markers and the corresponding vertebral landmarks respectively. The close relation between the position of the individual skin marker and its corresponding landmark on the vertebrae is conserved through all postures (regression coefficients: 0.7200.6 mug/kg/min was predictive of mortality in 76% of the patients. We conclude that the prognosis in children requiring renal replacement therapy depends on the severity of the clinical state at the time of starting therapy, principally on the hemodynamic situation. PMID- 16047226 TI - Hypercalciuria and recurrent urinary tract infections: incidence and symptoms in children over 5 years of age. AB - Hypercalciuria is an important and common risk factor in the formation of renal stones. In this study we evaluated the incidence and the clinical presentation of hypercalciuria in 75 children over 5 years of age with the diagnosis of recurrent urinary tract infection (UTI). We measured random urinary calcium/creatinine value (three times), 24-h urinary calcium excretion, serum calcium, phosphorus, electrolytes, blood gas, blood urea nitrogen and creatinine levels. Hypercalciuria was found in 32 patients (43%). The mean urinary calcium/creatinine ratio for hypercalciuric patients was 0.50+/-0.21 mg/mg (min: 0.24, max: 2.60). The mean urinary calcium/creatinine ratio for the rest of the study population--those without hypercalciuria--was 0.10+/-0.04 mg/mg (min: 0.01, max: 0.18). Presenting symptoms of the hypercalciuric patients and normocalciuric patients were similar. History of familial urolithiasis was positive in 19 patients (59%). Predisposing urinary tract abnormalities in recurrent UTI was shown in 12 of the hypercalciuric patients (12/32, 37.5%) and 8 of the normocalciuric patients (8/43, 19%) without a statistically significant difference between. We conclude that hypercalciuria is not a rare finding among recurrent UTI cases in Turkish children. Hypercalciuria does not modify the clinical presentation of UTI, and we suggest the investigation of urinary calcium excretion in children with recurrent UTI. PMID- 16047228 TI - American Chemical Society--220th National Meeting. Division of medicinal chemistry--selected symposia. 20-24 August 2000, Washington DC, USA. PMID- 16047227 TI - The aluminum content of bone increases with age, but is not higher in hip fracture cases with and without dementia compared to controls. AB - Aluminum is considered a potentially toxic metal, and aluminum poisoning may lead to three types of disorders: aluminum-induced bone disease, microcytic anemia and encephalopathy. This is well known in patients with chronic renal failure, but since healthy subjects with normal renal function retain 4% of the aluminum consumed, they are also at risk of long-term low-grade aluminum intoxication. Included in this study were a total of 172 patients (age range 16-98 years) with the aim of examining whether aluminum accumulates in bone with increasing age. Additionally, we aimed to investigate whether the aluminum content of bone differs between controls and hip fracture cases with and without dementia, in particular in those with Alzheimer's disease. During operations for all cases, bone biopsies were taken with an aluminum-free instrument from the trabecular bone. The samples were measured for their content of aluminum using an inductively coupled mass spectrometer. We found an exponential increase in aluminum content of bone with age. The average aluminum values, adjusted for age, were similar in men and women (P=0.46). No significant differences in sex- and age-adjusted mean aluminum values between the controls and the hip fracture cases with (P=0.72) and without (P=0.33) dementia could be detected. The average aluminum concentration among cases with Alzheimer's disease was also similar to the values of hip fracture patients with other types of dementia (P=0.47). Odds ratios of hip fracture for each quartile of aluminum content in bone were also estimated to detect non-linear effects, but we did not find any statistically significant association remaining after age and sex adjustment. Thus, our results indicate that we accumulate aluminum in bone over our life span, but this does not seem to be of major pathogenetic significance for the occurrence of hip fracture or dementia. PMID- 16047229 TI - Division of Medicinal Chemistry--general oral papers. PMID- 16047231 TI - Division of Medicinal Chemistry--P2 nucleotide receptors. PMID- 16047230 TI - Division of Medicinal Chemistry--poster session. PMID- 16047233 TI - Division of Medicinal Chemistry--integrins: past, present and future. PMID- 16047232 TI - Division of Medicinal Chemistry--small molecule inhibitors of protein-protein interactions. AB - Protein-protein interactions lie at the heart of the majority of cell recognition and signal transduction processes. Increasingly, medicinal chemistry researchers are interested in small molecule inhibitors of such interactions. This contrasts with approaches based on the design of small molecule receptor ligands mimicking physiological ligands, eg, neurotransmitters and small hormones. However, there have been significant successes, and two symposia at the 220th ACS meeting were devoted to this theme. One half-day symposium focused on general issues of protein-protein interaction, and a whole-day symposium focused on the integrin family of proteins (reviewed by Giancotti and Ruoslahti, Science (1999) 285:1028). PMID- 16047234 TI - Division of Medicinal Chemistry--chemoprevention symposium. PMID- 16047236 TI - Division of Medicinal Chemistry--cardiovascular symposium. PMID- 16047235 TI - Division of Medicinal Chemistry--dual acting drugs. PMID- 16047237 TI - Division of Carbohydrate Chemistry--Inosine Monophosphate Dehydrogenase: Perspectives on a Major Therapeutic Target. PMID- 16047238 TI - Division of Organic Chemistry--general oral papers. PMID- 16047239 TI - Division of Organic Chemistry--combinatorial and solid phase chemistry. PMID- 16047240 TI - Division of Organic Chemistry--technical achievements in organic chemistry and asymmetric synthesis symposia. PMID- 16047241 TI - Divisions of Chemical Information and Computers in Chemistry--symposia. PMID- 16047244 TI - Transplantation, pharmacogenomics and poster highlights. PMID- 16047242 TI - Biotechnology 2000: 11th International Biotechnology Symposium and Exhibition. PMID- 16047245 TI - Cell interactions. PMID- 16047246 TI - General presentations. PMID- 16047247 TI - Healthcare, molecular tools and applied genome research. AB - Biotechnology 2000 offered a rare opportunity for scientists from academia and industry to present and discuss data in fields as diverse as environmental biotechnology and applied genome research. The healthcare section of the meeting encompassed a number of gene therapy delivery systems that are successfully treating genetic disorders. Beta-thalassemia is being corrected in mice by continous erythropoeitin delivery from engineered muscles cells, and from naked DNA electrotransfer into muscles, as described by Dr JM Heard (Institut Pasteur, Paris, France). Dr Reszka (Max-Delbrueck-Centrum fuer Molekulare Medizin, Berlin, Germany), meanwhile, described a treatment for liver metastasis in the form of a drug carrier emolization system, DCES (Max-Delbrueck-Centrum fuer Molekulare Medizin), composed of surface modified liposomes and a substance for chemo occlusion, which drastically reduces the blood supply to the tumor and promotes apoptosis, necrosis and antiangiogenesis. In the molecular tools section, Willem Stemmer (Maxygen Inc, Redwood City, CA, USA) gave an insight into the importance that techniques, such as molecular breeding (DNA shuffling), have in the evolution of molecules with improved function, over a range of fields including pharmaceuticals, vaccines, agriculture and chemicals. Technologies, such as ribosome display, which can incorporate the evolution and the specific enrichment of proteins/peptides in cycles of selection, could play an enormous role in the production of novel therapeutics and diagnostics in future years, as explained by Andreas Pluckthun (Institute of Biochemistry, University of Zurich, Switzerland). Applied genome research offered technologies, such as the 'in vitro expression cloning', described by Dr Zwick (Promega Corp, Madison, WI, USA), are providing a functional analysis for the overwhelming flow of data emerging from high throughput sequencing of genomes and from high-density gene expression microarrays (DNA chips). The importance of bioinformatics was stressed throughout the conference. In particular, its applications in the storage, analysis and visualization of data, but also the linkage of databases and data mining tools. It is the storage and linkage that generates insight into disease processes, enabling the nomination of candidate drug targets. PMID- 16047248 TI - The European Association of Pharma Biotechnology Satellite Meeting. PMID- 16047249 TI - Vascular biology--XIth International Meeting. 5-9 September 2000, Geneva, Switzerland. PMID- 16047250 TI - Drug Discovery Technology 2000--IBC's Fifth Annual Congress. 14-17 August 2000, Boston, MA, USA. PMID- 16047251 TI - Rational Approaches to Drug Design: Quantitative Structure-Activity Relationships -13th European Symposium. 27 August-1 September 2000, Dusseldorf, Germany. PMID- 16047252 TI - Carbohydrate Symposium--20th International Meeting. 27 August-1 September 2000, Hamburg, Germany. AB - The symposium was attended by experts of carbohydrate science and encompassed a wide spectrum of carbohydrate chemistry and biology. Data presented included novel methods of synthesizing carbohydrates, as well as the identification and elucidation of new carbohydrates. Clinical aspects of the symposium concentrated on carbohydrate vaccines, influenza treatment and diagnosis, and antidiabetics. PMID- 16047253 TI - Production site transfer: successful approaches to overcoming technical, regulatory and compliance challenges. 22-23 August 2000, San Francisco, CA, USA. AB - The conference brought together a number of process professionals, who discussed their insights on the challenges presented in transferring a process from one production site to another. It was emphasized that this transfer could occur either within a company (ie, pilot plant to full-scale manufacturing) or between companies (ie, to a contract manufacturer). The transfer process requires not only the transfer of production, but also the transfer of analytical methods. While the primary focus of the presenters was on biologics transfer, the subject matter was equally applicable to chemical processes. PMID- 16047254 TI - Recent advances in cytokine synthesis inhibitors. PMID- 16047255 TI - Recent progress in protein farnesyltransferase inhibition. AB - This review aims to provide a recent update on protein farnesyltransferase inhibition, covering the period 1999 to mid 2000. A brief overview of the project rationale and its most recent evolution is also presented. For each inhibitor series, the most advanced molecules are presented along with a short description of the patent environment, focusing on second generation patents that cover new applications for protein farnesyltransferase inhibitors (PFTIs). PMID- 16047256 TI - Effects of VEGF on hemodynamics and cardiac function: characterization and mechanisms. AB - Vascular endothelial growth factor (VEGF), a mitogen specific for endothelial cells, produces beneficial angiogenesis in animal models of ischemic diseases; however, its side effects on hemodynamics and cardiac function may limit clinical use for some indications. This review focuses on the VEGF-induced hemodynamic profile, including hypotension, tachycardia, decreases in cardiac output and stroke volume, and changes in preload and afterload. The hemodynamic effects of VEGF are due to vasodilation and vascular hyperpermeability, which are mainly mediated by nitric oxide. Finally, the regimen or strategy for attenuation or prevention of these side effects is discussed. PMID- 16047257 TI - Clamikalant (Aventis). AB - Clamikalant is a cardioselective blocker of the ATP-dependent potassium channel (KATP) which is under development by Aventis Pharma (formerly Hoechst Marion Roussel) for the potential treatment of cardiac arrhythmia. The sodium salt, HMR 1098, is in phase II trials. Aventis plans for an iv preparation of the drug to be launched in 2004, and an oral preparation to be available in 2005. Clamikalant prevented ischemia-induced reductions in refractory period in dogs with ventricular fibrillation without significant hemodynamic effects or alteration in blood glucose levels. HMR-1883 exerted an anti-arrhythmic effect in a model of isolated hearts from male White New Zealand rabbits, and indicated and did not interfere with post-ischemic hyperemia. PMID- 16047258 TI - Vapreotide (Debipharm). AB - Debiopharm, under license from Tulane University, is developing vapreotide, a somatostatin analog, for the potential treatment of cancer, GI disorders and bleeding. Phase III trials have been initiated in France, investigating the utility of vapreotide in the treatment of prostate cancer. Phase III trials are also underway in France and Asia in patients with acute bleeding from esophageal varices. Debiopharm is also investigating the use of vapreotide in the treatment of acromegaly, gastrointestinal fistulae, AIDS-related and chemotherapy-induced diarrhea, and various neuroendocrine tumors. Vapreotide may also be useful for inducing hemostasis in cases of acute hemorrhage of the upper gastrointestinal tract. PMID- 16047259 TI - ADL-2-1294 (Adolor). AB - Adolor is developing various local and topical formulations of the antidiarrheal compound loperamide (ADL-2-1294), which acts as an opiate receptor agonist in the peripheral nervous system, for the potential treatment of pruritus and pain associated with burns, wounds, eye diseases and inflammation. Both the topical formulation and ophthalmic formulation are in phase II clinical trials. TOPICAL FORMULATION: A topical formulation is being developed for pain associated with cutaneous inflammatory lesions and other indications associated with inflammatory pain. Phase I trials were completed in spring 1997. The trials assessed the safety, tolerance, pharmacokinetics, and topical sensitivity of ADL-2-1294 in 35 burn patient volunteers. Following positive results, Adolor began two dose ranging phase II trials in patients with minor burns, abrasions and sunburn in the first half of 1997. In July 1997, Adolor was to initiate the preclinical development of ADL-2-1294 for the potential treatment of hyperalgesia associated with surgical wounds. OPHTHALMIC FORMULATION: By July 1999, phase II clinical trials of ADL-2-1294 for the potential treatment of inflammatory corneal pain were underway. In June 1998, the FDA accepted an IND for ADL-2-1294 for the treatment of inflammatory pain associated with corneal abrasions, surgical and laser keratectomies and keratoconjunctivitis. By this time, a phase I study had been initiated to assess safety and efficacy parameters. By July 1997, an ophthalmic formulation was in preclinical development for the treatment of corneal hyperalgesia. In preclinical studies with the compound, efficacy was demonstrated in animal models of inflammatory pain. OTHER FORMULATIONS: By 1999, Adolor was also investigating mucosal, post-surgical and intra-articular indications for the compound. By 1996, Adolor had plans to carry out clinical development alone, up to and including phase II trials. Then, the company were to seek a corporate partner, probably from Japan, and hold on to US and European rights. Beyond phase III trials, Adolor had expected to sell the compound to a major US or European company. In 1997, Adolor licensed ADL-2-1294 to the South Korean OTC topically-administered epidermal analgesic. In July 1999, Adolor licensed worldwide (excluding Korea) prescription and OTC development and commercial rights to topical dermal ADL-2-1294, for use in the treatment of inflammatory pain, itching and other undisclosed indications, to SmithKline Beecham. The company was issued US-05849761 and US-05849762 in January 1999 covering the use of antidiarrheal opioid compounds for the treatment of inflammatory pain and the treatment of pruritus. PMID- 16047260 TI - Homocysteine, B-vitamins, and the risk of cardiovascular disease. PMID- 16047261 TI - Homocysteine: overview of biochemistry, molecular biology, and role in disease processes. AB - Homocysteine is derived from the essential amino acid methionine and plays a vital role in cellular homeostasis in man. Homocysteine levels depend on its synthesis, involving methionine adenosyltransferase, S-adenosylmethionine dependent methyltransferases such as glycine N-methyltransferase, and S adenosylhomocysteine hydrolase; its remethylation to methionine by methionine synthase, which requires methionine synthase reductase, vitamin B (12), and 5 methyltetrahydrofolate produced by methylenetetrahydrofolate reductase or betaine methyltransferase; and its degradation by transsulfuration involving cystathionine beta-synthase. The control of homocysteine metabolism involves changes of tissue content or inherent kinetic properties of the enzymes. In particular, S-adenosylmethionine acts as a switch between remethylation and transsulfuration through its allosteric inhibition of methylenetetrahydrofolate reductase and activation of cystathionine beta-synthase. Mutant alleles of genes for these enzymes can lead to severe loss of function and varying severity of disease. Several defects lead to severe hyperhomocysteinemia, the most common form being cystathionine beta-synthase deficiency, with more than a hundred reported mutations. Less severe elevations of plasma homocysteine are caused by folate and vitamin B (12) deficiency, and renal disease and moderate hyperhomocysteinemia are associated with several common disease states such as cardiovascular disease. Homocysteine toxicity is likely direct or caused by disturbed levels of associated metabolites; for example, methylation reactions through elevated S-adenosylhomocysteine. PMID- 16047262 TI - Folate metabolism and cardiovascular disease. AB - Folate is a water-soluble vitamin that occurs in different chemical forms distinguished by their oxidation state and the specific type of one-carbon substitution. Folates occur in natural food sources as reduced methylated or formylated tetrahydrofolate. Folic acid is a synthetic analogue with no metabolic activity of its own. Pharmacological doses of folic acid cause it to appear in plasma, where it has unknown, but potentially adverse, effects. This review discusses folate absorption, body distribution, and intracellular folate metabolism. The main physiological functions of folate can be classified as methylation and DNA synthesis. Several mechanisms act in concert to regulate the folate metabolic pathways to ensure that both functions of folate are fulfilled properly. B-vitamin deficiencies and genetic polymorphisms (particularly the C677T mutation in the methylenetetrahydrofolatereductase gene) have multiple effects on folate metabolism. Impairment of the methylation cycle, for example, leads to hyperhomocysteinemia, a proposed atherothrombotic factor. However, methylation disturbances also result in hypomethylation of DNA and other molecules, which may also contribute to the pathogenesis of cardiovascular disease. As cardiovascular researchers, we should try to develop a more integrative view on folate metabolism, rather than focusing merely on hyperhomocysteinemia. PMID- 16047263 TI - Genetic determinants of plasma total homocysteine. AB - Hyperhomocysteinemia (Hhcy) is an established risk factor for various pathologies including arterial vascular disease and venous thrombosis, congenital malformations and other pregnancy complications, and dementia. Homocysteine remethylation, transsulfuration, and export to the blood/extracellular compartment determine homocysteine concentrations. Any disturbance in these routes may lead to Hhcy and potentially increase risk of disease. In this report, we aim to review all known polymorphisms involved in homocysteine and B-vitamin metabolism that have been assessed for their effect on tHcy. In the last section, we summarize the polymorphisms, for which the obtained data provides evidence for their involvement in Hhcy at the population level, and discuss how to continue our search for genetic determinants of tHcy. PMID- 16047264 TI - Dietary determinants of plasma homocysteine concentrations. AB - Severe hyperhomocysteinemia is typically caused by rare enzymatic defects or by renal failure. In contrast, mild to moderate hyperhomocysteinemia chiefly results from suboptimal status of nutritional factors involved in homocysteine metabolism. Low dietary intake of folate is the most important nutritional cause of elevated homocysteine (tHcy) concentrations. Folic acid is more effective than dietary folate in lowering tHcy concentrations, and a daily dose of 400 mug of folic acid is the minimum daily dose associated with the maximum tHcy-lowering effect ( approximately 20-25% reduction). Mean fasting tHcy concentrations have dropped substantially in populations with mandatory folic acid fortification, and other B-vitamins, such as vitamin B (12), are important determinants of tHcy levels in this setting. Vitamins B (2) and B (6) have little influence on fasting tHcy concentrations, although the former may be relevant in individuals with the MTHFR 677 TT-genotype, and the latter may improve tHcy catabolism in elderly individuals. Betaine and choline can lower fasting tHcy concentrations to a similar extent as folic acid, particularly in the setting of a high intake of methionine. Consumption of tea and coffee increase tHcy concentrations by up to 20%. A high-protein meal also increases tHcy, but these changes are transient, and levels return to normal after an overnight fast. Serine and cystine also influence the methionine-induced postprandial rise in tHcy concentrations. In conclusion, alteration in dietary intake or use of folic acid supplements can substantially lower tHcy concentrations. However, it is not known whether lowering tHcy levels can reduce the risk of cardiovascular disease or cognitive decline or prevent pregnancy complications or osteoporosis. PMID- 16047265 TI - Effect of drugs on homocysteine concentrations. AB - Many studies conducted over the last two decades have shown that drug treatment for common medical conditions may have an adverse effect on plasma total homocysteine (tHcy) concentrations. The mechanism for the effects of individual drugs on tHcy concentrations is frequently unknown, as the mechanism of action of the drug may not be established, or the drug is typically administered in combination with other drugs. Some drugs are believed to alter tHcy concentrations by interfering in the metabolism of folate or vitamins B (12) or B (6) or by altering renal function, but the underlying mechanisms for the effects on tHcy concentrations of many drugs remains to be discovered. Several widely used drugs, such as lipid-lowering drugs (like fibrates and niacin) or oral hypoglycemic drugs (like metformin), insulin, drugs used in rheumatoid arthritis, and anticonvulsants cause elevated tHcy concentrations. Sex hormones have variable effects on tHcy levels, and N-acetylcysteine lowers tHcy. The mechanisms of action of drugs on tHcy concentrations and strategies to avoid tHcy elevation have been studied. Assuming that the association of tHcy with cardiovascular disease is causal, this article focuses on the adverse effect on tHcy levels of fibrates, statins and niacin, antihypertensive drugs, metformin, methotrexate and sulfasalazine, anticonvulsant drugs, and levodopa and reviews strategies to avoid such effects. The clinical significance, if any, of these adverse effects on plasma tHcy concentrations remains to be determined. PMID- 16047266 TI - Novel and established markers of cobalamin deficiency: complementary or exclusive diagnostic strategies. AB - New developments in diagnostic markers and a better understanding of the limitations of traditional diagnostic strategies have allowed diagnosis of earlier stages and atypical forms of cobalamin deficiency. Still, there are no generally accepted guidelines for the definition, diagnosis, treatment, and follow-up of cobalamin deficiency. The new trend toward defining cobalamin deficiency purely on the basis of biochemical test outcomes in the absence of overt clinical signs and symptoms could, however, be problematic and may result in overdiagnosis and overtreatment. Use of metabolic markers for the assessment of cobalamin deficiency allows the demonstration of tissue deficiency, but the establishment of the cause of deficiency should also be part of the diagnostic approach. Four groups of diagnostic tests are currently available and these include total cobalamin and cobalamin fractions (such as holo-transcobalamin), tests of gastrointestinal dysfunction, tests of metabolic function, and different gene tests. Among the available tests, only homocysteine, methylmalonic acid, holo-transcobalamin, and possibly methylcitric acid are considered to be useful in clinical practice to add to cobalamin. Gastrointestinal function tests may identify the cause of cobalamin deficiency, whereas the diagnostic usefulness of genetic testing needs to be evaluated. This article provides an overview of recent developments and a reappraisal of novel and established diagnostic markers for cobalamin deficiency. PMID- 16047267 TI - Case for folic acid and vitamin B12 fortification in Europe. AB - The number of pregnancies affected by neural tube defects has been estimated to be 4000/year in Europe, with a higher prevalence in Celtic populations and in women of low socioeconomic status. Since the 1980s, it has been shown that supplementation with folic acid during the periconceptual period reduces the risk of neural tube defects in the fetus. However, in view of the period during which supplementation should be taken (< 4 weeks before conception until 8-10 weeks after) and the fact that in some countries 30-50% of pregnancies are unplanned, a public health initiative based solely on increasing dietary folate intake or recommendations on use of folic acid supplements is likely to be insufficient. Mandatory fortification has been started in 38 countries throughout the world. Several European countries have advocated mandatory flour folic acid fortification over the last 6 years, but none has introduced it. A recent public health decision in Hungary stimulated flour fortification on a voluntary basis, but it remains the only European country to take this action. Many European countries have deferred a decision to introduce fortification because of concerns about possible masking of vitamin B (12) deficiency. This review advocates a proposal for combined fortification of folic acid and vitamin B (12) to address possible hazards of fortification with folic acid alone. PMID- 16047268 TI - Mechanisms of the atherogenic effects of elevated homocysteine in experimental models. AB - Hyperhomocysteinemia is a risk factor for cardiovascular disease and stroke. During the last decade, considerable progress in delineating the mechanisms that underlie the atherogenic effects of hyperhomocysteinemia has been achieved through the use of experimental animal models. Among the most informative animal models are those that use genetic and dietary approaches to produce hyperhomocysteinemia in mice. Recent findings demonstrate that hyperhomocysteinemia can accelerate the development of atherosclerosis in susceptible models such as the apolipoprotein E-deficient mouse. Hyperhomocysteinemia also is a potent inducer of endothelial dysfunction, particularly in small vessels such as cerebral arterioles. Mechanisms of endothelial dysfunction may include inhibition of endothelial nitric oxide synthase by its endogenous inhibitor, asymmetric dimethylarginine, and oxidative inactivation of nitric oxide mediated by upregulation of prooxidant enzymes and downregulation of antioxidant enzymes. There also is good evidence from animal models that hyperhomocysteinemia produces endoplasmic reticulum stress, which may contribute to atherosclerosis and endothelial dysfunction by activating signal transduction pathways leading to inflammation, oxidative stress, and apoptosis. PMID- 16047269 TI - Homocysteine and endothelial function in human studies. AB - The endothelium plays a key role in the pathophysiology of vascular disease. Impaired flow-mediated dilatation (FMD) is a measure of endothelial dysfunction resulting from reduced bioavailability of nitric oxide (NO). Patients with homocystinuria manifest with impaired FMD, but in mild hyperhomocysteinemia, the evidence is conflicting. Oral loading with methionine or homocysteine impairs FMD, but it remains unproven that this effect is mediated directly by homocysteine. In addition, there is no clear consensus as to a mechanisms by which homocysteine would induce endothelial dysfunction. Folate administration lowers plasma homocysteine and enhances FMD. However, the effect of folate only appears to occur at high doses and with a time course that would indicate that it is acting by a mechanism independent of homocysteine lowering. It is possible that folate, in pharmacological doses, may enhance the NO activity by influencing NO-tetrahydrobiopterin interactions. These studies provide some insights and raise intriguing questions concerning the relationship between homocysteine, folate, and endothelial function. However, changes in FMD may not translate into vascular endpoints, and the outcomes of clinical intervention trials with different doses of folic acid are awaited with interest. PMID- 16047270 TI - Homocysteine and venous thrombosis. AB - Elevated plasma total homocysteine concentration is a risk factor for venous thrombosis. The association is well established in patients with homocystinuria irrespective of the genetic etiology and metabolic background. Homocystinuria is a human model of chronic exposure to very high concentrations of plasma homocysteine and reflects an abnormal amino acid metabolism. Elevated homocysteine levels in patients with venous thrombosis have attracted considerable interest because homocysteine is a potentially reversible thrombophilic marker for venous thrombosis. In contrast to homocystinuria, hyperhomocysteinemia is mild and reflects environmental and constitutional factors such as age, intake of B-vitamins, derangements of metabolism, and renal impairment. This review examines the evidence for the relationship of homocysteine with risk of venous thrombosis in homocystinuria and in the general population. PMID- 16047271 TI - Homocysteine and folic acid: implications for pregnancy. AB - The role of folic acid and homocysteine in pregnancy is becoming clearer. The efforts of many countries to prevent neural tube defects through public awareness of folic acid have been disappointing, but evidence is now emerging that the food fortification programs in the United States and Canada are effective in reducing the numbers of neural tube defects, and there may be additional benefits in terms of other congenital defects such as oral-facial clefts and congenital heart disease. Homocysteine has a significant association with vascular disease in later life, is elevated in preeclampsia, and has been associated with other pregnancy complications such as early pregnancy loss. The data from cohorts of women with a history of preeclampsia during pregnancy indicate that they are at increased risk for cardiovascular and cerebrovascular disease in later life. Elevated homocysteine concentrations may be a common link that accounts for these associations. PMID- 16047272 TI - Homocysteine and methionine metabolism in renal failure. AB - Renal insufficiency is invariably accompanied by elevated plasma concentrations of the sulfur-containing and potentially vasculotoxic amino acid homocysteine. There is a strong relationship between glomerular filtration rate and plasma homocysteine concentration. Unlike creatinine, however, homocysteine is avidly reabsorbed in the renal tubules, and its urinary excretion is minimal. There is no evidence that homocysteine is actively removed by the human kidney. In renal insufficiency, plasma concentrations of S-adenosylmethionine, S adenosylhomocysteine, cystathionine, cysteine, and sulfate are elevated, pointing to a remethylation or distal transsulfuration/oxidation block as the cause of hyperhomocysteinemia in renal failure. Stable isotope techniques have shown that both whole-body homocysteine remethylation and methionine transmethylation are decreased in renal failure, whereas homocysteine transsulfuration seems intact. Metabolic homocysteine clearance (i.e., transsulfuration relative to plasma homocysteine) is decreased to a major extent. These metabolic disturbances in renal failure can only be partially restored with current treatments. Folic acid treatment lowers plasma homocysteine concentration and increases remethylation and transmethylation rates. Plasma homocysteine, however, is not normalized, and metabolic homocysteine clearance by transsulfuration remains impaired. According to the currently available data, effective normalization of plasma homocysteine can only be obtained when its metabolic clearance through transsulfuration is restored. PMID- 16047273 TI - Homocysteine and cognitive function. AB - The prevention and treatment of age-related cognitive impairment and dementia is one of the greatest and most elusive challenges of our time. The prevalence of dementia increases exponentially with age, as does the prevalence of those with micronutrient deficiency. Several studies have shown that elevated homocysteine is correlated with cognitive decline and with cerebral atrophy and that it predicts the subsequent development of dementia in cognitively intact middle-aged and elderly individuals. If elevated homocysteine promotes cognitive dysfunction, then lowering homocysteine by means of B-vitamin supplementation may protect cognitive function by arresting or slowing the disease process. PMID- 16047274 TI - Homocysteine-lowering trials for prevention of heart disease and stroke. AB - Dietary supplementation with B-vitamins that lower plasma total homocysteine concentrations are expected to lower the risk of cardiovascular disease. Folic acid and vitamin B (12) lower blood homocysteine concentrations by about 25-30% in populations without folic acid fortification, but by only 10-15% in populations with fortification. In observational studies, 25% lower homocysteine is associated with about 10% less coronary heart disease (CHD) and about 20% less stroke. This review examines the current status of 12 large-scale randomized trials of B-vitamin supplementation and risk of cardiovascular disease. Seven of these trials are being performed in populations without fortification (five involving participants with prior CHD, two with prior stroke) and five in populations with fortification (two with prior CHD, two with renal disease, and one with prior stroke). Many of these trials may not have included a sufficient number of people or lasted long enough to have adequate power to exclude false negative results. Taken together, however, these trials involve 32,000 patients with prior vascular disease in unfortified populations and 20,000 (14,000 with vascular disease and 6000 with renal disease) patients in fortified populations. A metaanalysis of these trials should have adequate power to determine whether homocysteine-lowering vitamin supplements can reduce the risk of cardiovascular disease. PMID- 16047275 TI - [Exposure to particles an human health -- what has to be done?]. PMID- 16047278 TI - [Self assessment quality of life and lung diseases (SAQOL in patients with pulmonary carcinoma: influence to survival and impact of chronic obstructive pulmonary disease]. AB - BACKGROUND: Lung cancer and chronic obstructive pulmonary disease (COPD) often occur concomitantly. Whilst there are several questionnaires relating to COPD that have been translated into German (CCOPDQ, CRQ, SGRQ) there is a need for a short instrument measuring the impact of the concomitant diseases or intervention on a patient's Quality of Life specifically in German language. As there is to date no genuine German questionnaire neither in general, nor in detail for both, lung cancer and the often concomitant COPD, we created and tested an appropriate single instrument. METHODS: The feasibility and validity of a short, self administered quality of life questionnaire for use with patients diagnosed with COPD and/or pulmonary malignancies was evaluated. The 27 questions of Self Assessment Quality of Life in Lung Diseases (SAQOL) questionnaire were issued to out-patients with lung cancer (18 females, 75 males, mean 62.6 years, 18 to 81), 54 of with concomitant COPD. RESULTS: Correlations to comparable domains of the tumour-related QLQ-C30 of EORTC were good in cancer patients. Quality of life impact scores in the shorter and organ specific SAQOL domains showed even higher values than the QLQ C-30. A higher quote of Quality of life impaction was related to a poorer prognosis despite comparable tumour stages. There was a relevant impact related to the presence of COPD in addition to lung cancer. To investigate the validity in severe COPD an English version of the questionnaire was applied in the Respiratory Unit, Adelaide Australia, together with the Nottingham Health Profile in patients with long-term oxygen therapy (n = 20), which correlated well with the lung specific domains of SAQOL. CONCLUSION: We conclude that SAQOL is an easy to handle questionnaire, which shows on the one hand good correlation to the tumour-related domains of the broadly investigated and reliable QLQ C30, and on the other hand to the long-term used and valid lung specific generic Nottingham Health Profile in COPD. The important influence COPD in quality of life impact highlights the need for an organ specific questionnaire for both, lung cancer and obstructive airway disease. This important factor for quality of life alteration seems to be underestimated in other generic questionnaires or it needs the issue of at least two different and more time consuming instruments for adequate assessment. Additionally a high degree of quality of life impaction was linked to a short survival in our study patients with advanced lung cancer. PMID- 16047279 TI - [Primary pulmonary rhabdomyosarcoma as a rare differential diagnosis of small cell lung cancer]. AB - Primary pulmonary rhabdomyosarcoma is a rare entity and the histological differential diagnosis can be difficult. We report on a 43-year old female patient, smoker (25 pack-years), in whom a large solitary brain metastasis was diagnosed and enucleated. Histological examination revealed a typical small cell carcinoma and histological examination of biopsies obtained from a tumor in the left upper lobe of the lung was compatible with a small cell carcinoma. Despite chemotherapy there was a progressive tumor growth. Bronchial biopsies again showed a small cell tumor, although immunohistochemistry proved it to be a pleomorphic rhabdomyosarcoma. Due to the progressive tumor growth with necrosis and superinfection and a lack of further metastases lobectomy of the left upper lobe was performed, complicated by postoperative pleural empyema, limiting the possibilities of adjuvant therapy. Early relapse occurred with pleural, pulmonary, chest wall and spinal metastases. Laminectomy and extirpation of the spinal metastases, local radiotherapy and chemotherapy with iphosphamide and doxorubicine led to partial remission and clinical improvement for few months only. The patient died from metastatic primary rhabdomyosarcoma of the lung. This rare tumor mimicked small cell lung cancer. Appraisal of the atypical clinical course and a close dialogue between pathologists and clinicians enabled the correct diagnosis. PMID- 16047280 TI - [Drug transport in the respiratory epithelium]. AB - The aerosolic administration of peptidomimetic drugs with a peptide backbone may play a crucial role in the future treatment of diseases. Especially rational drug design offers an option to synthesize new drugs that are carried by specific drug transporters. Out of the presently identified transporter proteins PEPT1 and PEPT2, the high-affinity transporter PEPT2 is found in the respiratory tract. The transporter possess 12 membrane spanning domains and catalyses an electrogenic uphill drug transport using a transmembrane electrochemical proton gradient. PEPT2 is expressed in the bronchial epithelium and in alveolar type II pneumocytes in human airways. Kinetic studies demonstrated that peptidomimetic compounds including antibiotic, antiviral and antineoplastic drugs are carried by PEPT2. The transporter also carries delta-aminolevulinic acid into the airways. This molecule can be used for the diagnostics of pulmonary neoplasms and for photodynamic therapy. Using the recently published data on minimal structural requirements for PEPT2-substrates, rational drug design may lead to a new generation of respiratory drugs and prodrugs, which are delivered to the airways via the molecular mechanisms of the PEPT2 transport system. PMID- 16047281 TI - [Effects of particulate air pollution on human health. Statement of the German Society of Pneumology on the discussion about fine particulate air pollution]. PMID- 16047282 TI - [Imaging 2005 -- ultrasound of the chest]. PMID- 16047283 TI - [Broncho-alveolar lavage in humans and animals -- comparative aspects]. PMID- 16047284 TI - [Broncho-alveolar lavage in animals]. PMID- 16047285 TI - [Broncho-alveolar lavage in humans]. PMID- 16047286 TI - [Bronchoalveolar lavage in domestic animals]. PMID- 16047287 TI - [Broncho-alveolar lavage in dogs and cats--clinical aspects]. PMID- 16047288 TI - [broncho alveolar lavage in horses--clinical aspects]. PMID- 16047289 TI - [Broncho-alveolar lavage in animal experiments with specific regard to induced and therapeutic aspects in ARDS (acute respiratory distress syndromes) investigations]. PMID- 16047291 TI - Postexercise physiology and repeat performance behaviour of free-swimming smallmouth bass in an experimental raceway. AB - We studied postexercise physiology and behaviour of smallmouth bass (Micropterus dolomieu) that voluntarily ascended experimental raceways of varying length (20 50 m) against water velocities ranging from 8 to 120 cm/s. Our first objective was to link mean swimming speed to metabolism using patterns in postexercise muscle glycogen, muscle lactate, and plasma lactate. Our second objective was to examine several behavioural indices (attempt rate, success rate, and recovery time between an ascent and a subsequent attempt) and determine whether patterns in these data reflected those from the physiological measurements. Postexercise muscle glycogen and plasma lactate data suggest that smallmouth bass powered swimming speeds up to 70-80 cm/s using energy from aerobic processes. However, lactate did not begin to accumulate in the white muscle until speeds in excess of 120-130 cm/s were reached. The behavioural parameters measured did not indicate the presence of a physiological threshold at 70-80 cm/s; however, patterns in all factors changed appreciably when fish maintained speeds in excess of 120-130 cm/s. Therefore, it is clear that behaviour and physiology are tightly linked in this species and that maximum aerobic swimming capacity may not limit performance (or re-performance) during short-duration swims. PMID- 16047292 TI - Shifts in a fish's resource holding power during a contact paired interaction: the influence of a copper-contaminated diet in rainbow trout. AB - The influence of sublethal chronic dietary copper (Cu) exposure on the dominant subordinate relationship between pairs of juvenile rainbow trout (Oncorhynchus mykiss) was examined. Fish were fed either a normal (11 mg Cu kg(-1) food) or Cu contaminated (721 mg Cu kg(-1) food) diet for 8 wk. Paired interactions were observed--control versus control, Cu-exposed versus Cu-exposed, and control versus Cu-exposed fish--using a computer-aided video tracking system to measure duration of interactions, total distance moved, and the number of encounters during each contest. In concurrence with game theory, each interaction became escalated with a lesser size disparity between contestants. However, in Cu exposed versus Cu-exposed fish interactions, the dominant-subordinate relationship was decided sooner and with less aggression than a control versus control fish interaction with fish of a similar relative body mass disparity. During control versus Cu-exposed fish interactions, control fish would normally dominate interactions (12 out of 16 bouts) unless the Cu-exposed fish had a 15% body mass advantage. Muscle glycogen and lactate levels after each contest reflected the duration of bouts and winners of the contests, irrespective of Cu exposure. We conclude that Cu-contaminated fish are less able to compete and have lower resource holding power than controls and will withdraw from a contest at a lower level of interaction, unless a size advantage in the Cu-exposed fish increases the probability of winning. PMID- 16047293 TI - The integration of behaviour into comparative physiology. AB - Comparative physiology has traditionally focused on the physiological responses of animals to their physicochemical environment. In recent years, awareness has increased among physiologists of the potential for behavioural factors, such as the social environment of the animal, to affect physiological condition and responses. This recognition has led to an emerging trend within the field toward using multidisciplinary approaches that incorporate both behavioural and physiological techniques. Research areas in which the integrated study of behaviour and physiology has been particularly fruitful include the physiology of the social environment, sensory physiology and behaviour, and physiological constraints on behavioural ecology. The manner in which incorporating behavioural considerations has informed the physiological data collected is discussed for each of these areas using specific examples. PMID- 16047294 TI - [Clinical-therapeutic study of dementia in people with Down syndrome and the effectiveness of donepezil in this population]. AB - INTRODUCTION: People with Down syndrome have an early aging process, especially form their 40s. There is a significant average of them who initiate, from that age on, a progressive decline of their cognitive and functional abilities, due to a primary degenerative process Alzheimer's disease type. When assessing response to treatment with, pilot clinical trials on this population have demonstrated real benefits therapeutically. AIMS: To assess the efficacy and safety of a pharmacological treatment with donepezil over cognitive and behavioral disturbances on patients with Down syndrome older than 40 years, areas where family and professional educators of reference have observed cognitive and behavioral changes in comparison with their previous level of disability. PATIENTS AND METHODS: Patients have been selected from different institutions affiliated at the Catalan Foundation for the Down syndrome and by the Catalan Federation Pro Persons with Psychic Disability. Several deterioration, behavioral and functional assessment scales have been used, all of them validated into this population. RESULTS: The results of study demonstrated that donepezil slowed the progression of the cognitive dysfunction, especially during the first three months of treatment. This occurred for both cognitive and social-behavioral outcomes. CONCLUSIONS: Donepezil appears to be effective in the treatment of cognitive and behavioural disturbances associated with the progressive dementia syndrome in Down's. However, the sample sizes used in this, and all published studies are small and this emphasizes the need for a larger, multi-center trial to fully evaluate the nature and extent of the response of Down's syndrome patients to anticolinesterase therapy. PMID- 16047295 TI - [Spinal cord injury due to osteoarticular causes]. AB - INTRODUCTION: Spinal cord injury is one of the most tragic disabilities, because of its sequelae. Incidence ranges between 9 and 53 cases per million inhabitants. Injuries due to non-traumatic causes account for a significant number of cases but they have not been studied in depth. Our objective was to conduct a descriptive study of osteoarticular-type non-traumatic spinal cord injuries in our centre. PATIENTS AND METHODS: We carried out a retrospective study of the medical records of patients referred to our centre because of non-traumatic, degenerative spinal cord injury between 1998 and 2002. We evaluated age, sex, neurological injury classification according to the ASIA (American Spinal Injury Association) on admission and on discharge, complete (motor and sensory) or incomplete involvement, level of the injury and how it was caused. RESULTS: We found 365 records of patients with non-traumatic spinal injuries, 45 (12.3%) of which were osteoarticular-type injuries. The age group and most frequent cause were 56-69 years and canal stenosis, respectively. There was seen to be a predominance of males and incomplete injury. The neurological level that was most affected was the cervical spine. CONCLUSIONS: In our centre, the majority of cases of non-traumatic spinal cord injury due to osteoarticular causes are produced by canal stenosis, with a greater incidence in the cervical spine. It is more often observed in 56 to 69-year-old males and usually involves incomplete injuries. Such data match those obtained by other authors. PMID- 16047296 TI - [Frequency of neuropsychiatric signs and symptoms in patients with viral encephalitis]. AB - INTRODUCTION: Acute viral encephalitis (AVE) is a frequent condition that usually courses with psychiatric alterations but few systematic studies have been conducted to investigate it. AIMS: To determine the frequency and the progression of the neuropsychiatric symptoms in patients with AVE. PATIENTS AND METHODS: A retrospective study was carried out. AVE was defined as an acute and progressively coursing condition in previously healthy subjects, with clinical signs of diffuse alteration of the central nervous system, abnormal electroencephalogram and/or inflammatory cerebrospinal fluid (CSF). We excluded patients who previously had epilepsy, a positive serodiagnosis for human immunodeficiency virus (HIV), and cases compatible with herpes simplex encephalitis from electroencephalographic or imaging data with focalisation towards temporal, frontal, regions or a positive DNA test for herpes in CSF. Finally, 83 patients were included. The psychiatric signs and symptoms that were produced were recorded during the acute phase and one year after discharge from hospital (sequelae). RESULTS: The psychiatric disorders in the acute phase were psychomotor agitation (67%), drowsiness (55%), disorientation (47%), visual hallucinations (43%) and aggressiveness (34%). One year after hospitalisation, in a sample of 70 patients in a clinical control, we found memory disorders (16%), aggressiveness (9%), aphasia (8%), visual hallucinations (8%), and auditory hallucinations (7%). The mortality rate was 6%. CONCLUSIONS: Neuropsychiatric disorders are very frequent during the acute phase of viral encephalitis, which is relevant for the differential diagnosis in patients who visit emergency departments with behavioural disorders. One year after hospital discharge, the main sequelae are of a neuropsychiatric nature and cognitive impairment is predominant. PMID- 16047297 TI - [Hereditary neuromuscular diseases in paediatrics. Our experience over the last 14 years]. AB - INTRODUCTION: Hereditary neuromuscular diseases are disorders which can vary largely in their age of onset, symptoms and severity. Many are severe, disabling and have an important personal, familial and social impact and can restrict the prognosis for survival. The constant progress being made in diagnostics makes it necessary to continually update knowledge and information. PATIENTS AND METHODS: We carried out a review of the hereditary neuromuscular diseases contained in the Neuropaediatrics database at the Hospital Miguel Servet in Zaragoza from May 1990 to October 2004. RESULTS: Of the 7,805 patients in the database, 123 (1.5% of the total) were patients with hereditary neuromuscular diseases, of whom 71 were males and 52 females. These included: 35 sensory-motor hereditary neuropathies, 17 dystrophinopathies, 10 myotonic dystrophies, 10 spinal muscular atrophies, four merosin-deficient congenital dystrophies, four other muscular dystrophies, three mitochondrial myopathies, three myasthenias, two familial neuropathies with insensitivity to pain, two Friedreich's ataxias, one familial neuropathy with liability to pressure palsies, one case of Walker-Warburg syndrome, five polyneuropathies associated to leukodystrophy and another 25 cases that could not be classified. Genetic studies provided a diagnosis in 36 cases (29.2%): nine myotonic dystrophies, eight dystrophinopathies, eight cases of spinal muscular atrophy, four demyelinating sensory-motor hereditary neuropathies, two instances of Friedreich's ataxia, two limb-girdle muscular dystrophies, one congenital myasthenia, one McArdle's disease and one case of Kearns-Sayre syndrome. CONCLUSIONS: Genetic studies enable us to establish diagnoses that were previously limited to the realm of assumption, and allow us to avoid the need for muscle tissue biopsies, which is a welcome development, especially when dealing with children. Immunohistochemical studies need to be updated and biological samples should be systematically saved in cases where no diagnosis is reached. PMID- 16047298 TI - [Differential diagnosis of intracranial calcifications]. AB - INTRODUCTION: Intracranial calcifications are a frequent chance finding in many neuroimaging tests. We report two clinical cases with intracranial calcifications as a common feature, but which had very different aetiologies and presenting symptoms. CASE REPORTS: The first case involved a 51-year-old male with a single tonic-clonic convulsive seizure associated to hypocalcemia secondary to hypoparathyroidism who presented extensive brain calcifications affecting the dentate nuclei, pons, periependymal and basal ganglia, and whose convulsive attacks remitted once normal Ca2+ values were restored. The second case was a 25 year-old female patient whose convulsive seizures were yet another symptom of Fahr's syndrome, which was associated to spastic paralysis, athetosis, mental retardation and occasionally hypoplasia of the optic nerve; the calcifications were located in the grey nuclei of the cerebrum and cerebellum. CONCLUSIONS: A correct history and physical and neurological examination must be carried out, and a detailed study of Ca2+/ P+ and hormonal metabolism is also required. A CAT scan is the preferred neuroimaging technique to achieve a proper differential diagnosis and to decide on the therapeutic approach that is best suited to the pathophysiological mechanisms that lead to the formation of these calcium deposits. PMID- 16047299 TI - [Late onset epilepsy as the first symptom of pseudohypoparathyroidism]. AB - INTRODUCTION: The late onset epilepsy is defined like that start in the adult life after the 25 years old. The diagnostic assessment of the late onset epilepsy forces to discard, in the absence of the principal causes like stroke, tumors and trauma to the metabolic cause like plausible. The persistent hypocalcemia, apart of its acute repercussions, produces severe neurologic consequences. CASE REPORT: Feminine patient of 61 years old, that assists with a tonic clonic generalized seizure and tetany facts in the physical exam (Chvostek and Trousseau), round facies, obesity and metacarpals abnormalities. The hypocalcemia, hyperphosphatemia and resistance to parathyroid hormone (with normal levels of cAM in urine after stimulation with PTH) triad outline the pseudohypoparathyroidism. CONCLUSIONS: Consider by the clinical history, biochemical results and clinical findings consistent with Albright hereditary osteodystrophy a pseudohypoparathyroidism type Ia in this patient; is the more common form of this disorders and have a dominant pattern of inheritance, is produced by a detectable mutation in the stimulatory Gs alpha, one protein of the adenylil cyclase complex (GNAS1) that is traduced in a inefficacy of the PTH renal receptor. The main goals of treatment are reduce and maintain the serum calcium and PTH levels, respectively. PMID- 16047300 TI - [Sarcoid myopathy. Report of two cases and review of the bibliography]. AB - INTRODUCTION: 5% of patients with sarcoidosis are affected by neurological complications, of which myopathy is one of the least frequent. We report the clinical course and therapeutic response of two patients with sarcoid myopathy. CASE REPORTS: We observed two females aged 63 and 55 who were previously diagnosed with sarcoidosis, which in one case was cutaneous and ophthalmic and in the other pulmonary. Both were asymptomatic from a systemic point of view when they visited the Neurology department. Both patients developed chronic myopathy (with a history of 9 months and 6 years), which was predominantly proximal, painful and both of them had normal or slightly high levels of creatine phosphokinase. The EMG was clearly myopathic in the two patients. A muscle biopsy showed a mononuclear-cell infiltrate with the formation of non-caseating granulomas. Both patients were treated with prednisone, although the therapeutic response was different in each case. One of the patients showed an important improvement in just a few days, while in the other case the disease remained stable, despite adding, first, azathioprine and later methotrexate to the treatment. CONCLUSIONS: Sarcoidosis can cause chronic, predominantly proximal, myopathy, and chiefly affects females over 50 years of age. The therapeutic response to steroids and immunosuppressants varies from case to case. PMID- 16047301 TI - [Neurophysiological studies of the neuromuscular junction]. AB - INTRODUCTION: The role played by neurophysiological studies (NPS) in the diagnosis of diseases affecting neuromuscular transmission (NMT) is based on the study of the failure of muscle fibres to achieve a sufficient degree of depolarisation for the junction potential to reach the appropriate threshold and attain a muscular action potential. This totally or partially blocked impulse will give rise to different types of responses in neurophysiological tests. AIMS: To analyse the different NPS as diagnostic methods in diseases that affect NMT. DEVELOPMENT: The article offers a review of the concept of the safety margin at the neuromuscular junction and a description of the most common neurophysiological techniques currently in use--repetitive stimulation, as well as conventional or single fibre electromyography (EMG) with voluntary activation or axonal electrical activation. The most frequent findings in diseases affecting NMT are also discussed. CONCLUSIONS: NPS will be useful to confirm or reject the clinical diagnosis, to exclude other concomitant neuromuscular diseases, to establish whether the process is pre- or post-synaptic, to monitor the clinical course of the disease (when it is both natural or in response to the medical or surgical treatment) and also to enable the physician to determine the status of NMT in cases of clinical remission, as well as to detect subclinical disorders. Single fibre EMG studies are the most sensitive method of neurophysiological diagnosis when dealing with these diseases. PMID- 16047302 TI - [A proposal for a protocol for use in the evaluation of the executive functions]. AB - INTRODUCTION: Executive functions include a variety of components such as the capacity implicated in goal formulation, the faculties employed in processes planning, and the strategies used to achieve the pretended objectives. In a previous work, taking as starting basis those models which have attempted to clarify those processes implicated in executive functions, we posed an integrative model. DEVELOPMENT: Starting from this model, we now propose an assessment protocol. Thus, executive functions considered as problem solving, require in generic terms, objective selection, planning, and monitoring processes (tower of Hanoi and zoo map). Each of these sub-processes operate through the working memory both with the visospatial sketch and the phonological loop. The central executive system, or attentional supervisor system (ASS), acts when there is no known solution and we must create an alternative one. CONCLUSIONS: In this sense, the ASS could contain the following functions: amplification of the phonological loop and visospatial sketch capacity (Sternberg type tasks), information manipulation and actualization (n-back paradigm), information manipulation and maintenance (Wechsler Memory Scale letters and numbers), simultaneously work in two cognitive tasks (dual execution tasks), inhibition (Stroop and go-no go paradigms), and cognitive sets alternation (Wisconsin Card Sorting Test). Once this planning process has been done, we must take a decision (gambling task paradigm), being the somatic marker in charge of this process. PMID- 16047303 TI - [A study of the demand for electromyography in a general hospital]. PMID- 16047304 TI - [Bioelectrical impulse, synaptic and brain circuit quantifications, the origins of life and the main composition of neurons]. PMID- 16047306 TI - [Cerebrovascular disease as the presenting symptom of temporal arteritis]. PMID- 16047307 TI - A wide range of protein isoforms in serum and plasma uncovered by a quantitative intact protein analysis system. AB - We have implemented an orthogonal 3-D intact protein analysis system (IPAS) to quantitatively profile protein differences between human serum and plasma. Reference specimens consisting of pooled Caucasian-American serum, citrate anticoagulated plasma, and EDTA-anticoagulated plasma were each depleted of six highly abundant proteins, concentrated, and labeled with a different Cy dye (Cy5, Cy3, or Cy2). A mixture consisting of each of the labeled samples was subjected to three dimensions of separation based on charge, hydrophobicity, and molecular mass. Differences in the abundance of proteins between each of the three samples were determined. More than 5000 bands were found to have greater than two-fold difference in intensity between any pair of labeled specimens by quantitative imaging. As expected, some of the differences in band intensities between serum and plasma were attributable to proteins related to coagulation. Interestingly, many proteins were identified in multiple fractions, each exhibiting different pI, hydrophobicity, or molecular mass. This is likely reflective of the expression of different protein isoforms or specific protein cleavage products, as illustrated by complement component 3 precursor and clusterin. IPAS provides a high resolution, high sensitivity, and quantitative approach for the analysis of serum and plasma proteins, and allows assessment of PTMs as a potential source of biomarkers. PMID- 16047308 TI - Evaluation of prefractionation methods as a preparatory step for multidimensional based chromatography of serum proteins. AB - Prefractionations of proteins prior to their proteolysis, chromatography, and MS/MS analyses help reduce complexity and increase the yield of protein identifications. A number of methods were evaluated here for prefractionating serum samples distributed to the participating laboratories as part of the human Plasma Proteome Project. These methods include strong cation exchange (SCX) chromatography, slicing of SDS-PAGE gel bands, and liquid-phase IEF of the proteins. The fractionated proteins were trypsinized and the resulting peptides were resolved and analyzed by multidimensional protein identification technology coupled to IT MS/MS. The MS/MS spectra were clustered, combined, and searched against the IPI protein databank using Pep-Miner. The identification results were evaluated for the efficacy of the different prefractionation methodologies to identify larger numbers of proteins at higher confidence and to achieve the best coverage of the proteins with the identified peptides. Prefractionation based on SCX resulted in the largest number of identified proteins, followed by gel slices and then the liquid-phase IEF. An important observation was that each of the methods revealed a set of unique proteins, some identified with high confidence. Therefore, for comprehensive identification of the serum proteins, several different prefractionation approaches should be used in parallel. PMID- 16047309 TI - The human plasma proteome: analysis of Chinese serum using shotgun strategy. AB - We have investigated the serum proteome of Han-nationality Chinese by using shotgun strategy. A complete proteomics analysis was performed on two reference specimens from a total of 20 healthy donors, in which each sample was made from ten-pooled male or female serum, respectively. The methodology used encompassed (1) removal of six high-abundant proteins; (2) tryptic digestion of low- and high abundant proteins of serum; (3) separation of peptide mixture by RP-HPLC followed by ESI-MS/MS identification. A total of 944 nonredundant proteins were identified under a stringent filter condition (X(corr) > or = 1.9, > or = 2.2, and > or = 3.75, < or = C(n) > or = 0.1, and R(sp) > or = 4.0) in both pooled male and female samples, in which 594 and 622 entire proteins were found, respectively. Compared with the total 3020 protein identifications confirmed by more than one laboratory or more than one specimen in HUPO Plasma Proteome Project (PPP) participating laboratories recently, 206 proteins were identified with at least two distinct peptides per protein and 185 proteins were considered as high confidence identification. Moreover, some lower abundance serum proteins (ng/mL range) were detected, such as complement C5 and CA125, routinely used as an ovarian cancer marker in plasma and serum. The resulting nonredundant list of serum proteins would add significant information to the knowledge base of human plasma proteome and facilitate disease markers discovery. PMID- 16047310 TI - Efficient prefractionation of low-abundance proteins in human plasma and construction of a two-dimensional map. AB - Human plasma is the most clinically valuable specimen, containing not only a dynamic concentration range of protein components, but also several groups of high-abundance proteins that seriously interfere with the detection of low abundance potential biomarker proteins. To establish a high-throughput method for efficient depletion of high-abundance proteins and subsequent fractionation, prior to molecular analysis of proteins, we explored how coupled immunoaffinity columns, commercially available as multiple affinity removal columns (MARC) and free flow electrophoresis (FFE), could apply to the HUPO plasma proteome project. Here we report identification of proteins and construction of a human plasma 2-DE map devoid of six major abundance proteins (albumin, transferrin, IgG, IgA, haptoglobin, and antitrypsin) using MARC. The proteins were identified by PMF, matching with various internal 2-DE maps, resulting in a total of 144 nonredundant proteins that were identified from 398 spots. Tissue plasminogen activator, usually present at 10-60 ng/mL plasma, was also identified, indicative of a potentially low-abundance biomarker. Comparison of representative 2-D gel images of three ethnic groups (Caucasian, Asian-American, African-American) plasma exhibited minor differences in certain proteins between races and sample pretreatment. To establish a throughput fractionation of plasma samples by FFE, either MARC flow-through fractions or untreated samples of Korean serum were subjected to FFE. After separation of samples on FFE, an aliquot of each fraction was analyzed by 1-D gel, in which MARC separation was a prerequisite for FFE work. Thus, a working scheme of MARC --> FFE --> 1-D PAGE --> 2-D-nanoLC-MS/MS may be considered as a widely applicable standard platform technology for fractionation of complex samples like plasma. PMID- 16047311 TI - A simple and affordable method for high-throughput DNA extraction from animal tissues for polymerase chain reaction. AB - We have developed a very simple and inexpensive method for high-throughput DNA extraction from animal tissues. The procedure contains three steps (digestion, heating, and centrifugation) and it is compatible with the 96-well plate format commonly used in polymerase chain reaction (PCR) amplifications. The duration for processing a plate is about 1.5 h; therefore, one researcher can isolate DNA from up to 1000 samples during a single workday. A small piece of tissue (ca. 10-20 mg) yields enough template for at least 50-70 PCR amplifications, as demonstrated by using the processed samples as templates successfully for long distance PCR, multiplex PCR, and randomly amplified polymorphic DNA (RAPD) assay. The application of our method is expected to facilitate studies that require high throughput DNA isolation for PCR amplification, such as genotyping by microsatellites for mapping and genetic diversity studies, as well as mutant screening in zebrafish. PMID- 16047312 TI - Application of an external contactless conductivity detector for the analysis of beverages by microchip capillary electrophoresis. AB - Quantitative total ionic analysis of alcoholic and nonalcoholic beverages was performed by microchip capillary electrophoresis with external contactless conductivity detection. An electrolyte solution consisting of 10.5 mM histidine, 50 mM acetic acid, and 2 mM 18-crown-6 at pH 4.1 was used for the determination of NH(4) (+), K(+), Ca(2+), Na(+), and Mg(2+). Fast analysis of Cl(-), NO(3) (-), and SO(4) (2-) was achieved in 20 mM 2-(N-morpholino)ethanesulfonic acid /histidine electrolyte solution at pH 6.0 and the simultaneous separation of up to 12 inorganic and organic anions was performed in a solution containing 10 mM His and 7 mM glutamic acid at pH 5.75. Limits of detection ranged from 90 to 250 mug/L for inorganic cations and anions, and from 200 to 2000 mug/L for organic anions and phosphate. Calibration curves showed linear dependencies over one to two orders of magnitude when the stacking effect was minimized by injecting standard solutions prepared in background electrolyte solutions. Total analysis times of 35 and 90 s were achieved for the determination of 5 inorganic cations and for the simultaneous determination of 12 inorganic and organic anions, respectively, which represents a considerable reduction of analysis time compared to conventional separation methods used in food analysis. PMID- 16047313 TI - A fast method for quantitative proteomics based on a combination between two dimensional electrophoresis and 15N-metabolic labelling. AB - We provide a method for accurate protein quantitation that uses two-dimensional (2-D) gel electrophoresis for protein separation, but does not require extensive statistical analysis of staining intensities on gels. Instead, accurate quantitation occurs on the mass spectrometer (MAS) on multiple peptides to provide statistical evidence. In an example study, Sulfolobus solfataricus cells were grown on the carbon sources glucose, fructose and glutamate. The glucose phenotype (reference) was grown on (15)N-enriched medium. Next, the glutamate and the fructose phenotypes are mixed with the reference and two 2-D gels are created. Staining intensities of gel spots in this case are used for initial, semiquantitative assessment of differential expression. On this basis, spots are selected for accurate quantitation on the MAS. A number of differentially expressed proteins were found, for example: a (25.2 +/- 8.2)-fold upregulation of isocitrate lyase and a (7.14 +/- 0.82)-fold downregulation of glucose dehydrogenase on glutamate compared to glucose. With this protocol, intergel and interlaboratory comparisons are facilitated, since the light and heavy versions of a protein are equally affected by variations in sample preparation and buffer composition. Because the statistical evidence is gathered on the MAS, the need to run vast numbers of gels is removed. PMID- 16047314 TI - Poly(ethylene oxide) facilitates the characterization of an affinity between strongly basic proteins with DNA by affinity capillary electrophoresis. AB - In order to study kin17 protein-DNA affinity, we have developed a fast and reproducible capillary electrophoresis (CE) analysis of a strongly basic protein: kin17 protein, using a nonpermanent coating based on poly(ethylene oxide) (PEO) to avoid adsorption of kin17. The coating procedure was optimized to provide a residual and stable electroosmotic flow (EOF = 5 x 10(-5) cm(2)/V x s), exhibiting RSD of 0.3% and excellent long-term stability. Good intraday and interday reproducibility of kin17 migration times (0.8 and 0.3% relative standard deviation (RSD), respectively) enabled us to consider that the recovery percentage obtained for kin17 protein was satisfactory (79%). The potential of this PEO-based coating procedure was evaluated for affinity CE method in order to study the affinity of kin17 protein for two single-stranded DNA (ssDNA) models: polydeoxyadenylic acid and polydeoxycytidilic acid (pdA and pdC). Binding constants (1.5 x 10(7) +/- 17% and 1.7 x 10(7) + 25%M(-1)) were evaluated assuming a 1:1 affinity between kin17 and pdA or pdC, respectively. PMID- 16047315 TI - Solid-phase microextraction gas chromatography/mass spectrometry: a new method for species identification of truffles. AB - This study describes a rapid method to identify different truffle species by analysis of their volatile compound fraction using static headspace solid-phase microextraction gas chromatography/mass spectrometry. The volatile organic compounds (VOCs) were extracted using a new 2-cm 50/30 microm DVB/CAR/PDMS fiber placed for 10 min in the headspace of the truffle sample with the vial maintained at 20 degrees C (in a thermostatically controlled analysis room). The mass spectra of the VOC chromatograms were represented as 'fingerprints' of the analysed samples. Next, stepwise factorial discriminant analysis afforded a limited number of characteristic fragment ions that allowed a classification of the truffle species studied. This new method provides an effective approach to rapid quality control and identification of truffle species by analysis of their volatile fraction. Moreover, this method offers the advantage of minimizing thermal, mechanical, and chemical modifications of the truffles, thereby reducing the risk of analytical artifacts. PMID- 16047316 TI - Treatment methods for the determination of delta2H and delta18O of hair keratin by continuous-flow isotope-ratio mass spectrometry. AB - The structural proteins that comprise approximately 90% of animal hair have the potential to record environmentally and physiologically determined variation in delta2H and delta18O values of body water. Broad, systematic, geospatial variation in stable hydrogen and oxygen isotopes of environmental water and the capacity for rapid, precise measurement via methods such as high-temperature conversion elemental analyzer/isotope ratio mass spectrometry (TC/EA-IRMS) make these isotope systems particularly well suited for applications requiring the geolocation of hair samples. In order for such applications to be successful, however, methods must exist for the accurate determination of hair delta2H and delta18O values reflecting the primary products of biosynthesis. Here, we present the results of experiments designed to examine two potential inaccuracies affecting delta2H and delta18O measurements of hair: the contribution of non biologic hydrogen and oxygen to samples in the form of sorbed molecular water, and the exchange of hydroxyl-bound hydrogen between hair keratin and ambient water vapor. We show that rapid sorption of molecular water from the atmosphere can have a substantial effect on measured delta2H and delta18O values of hair (comprising approximately 7.7% of the measured isotopic signal for H and up to approximately 10.6% for O), but that this contribution can be effectively removed through vacuum-drying of samples for 6 days. Hydrogen exchange between hair keratin and ambient vapor is also rapid (reaching equilibrium within 3-4 days), with 9-16% of the total hydrogen available for exchange at room temperature. Based on the results of these experiments, we outline a recommended sample treatment procedure for routine measurement of delta2H and delta18O in mammal hair. PMID- 16047317 TI - Assessment of the plasma desorption time-of-flight mass spectrometry technique for pesticide adsorption and degradation on 'as-received' treated soil samples. AB - The assessment of the plasma desorption time-of-flight mass spectrometry (PD TOFMS) technique as a tool for direct characterization of pesticides adsorbed on agricultural soil is made for the first time in this study. Pellets of soils impregnated by solutions of three pesticides, namely norflurazon, malathion and oxyfluorfen, as well as deposits of these solutions onto aluminum surfaces, were investigated to this end. The yield values of the most characteristic peaks of the negative ion mass spectra were used to determine both the lowest concentrations detected on soils and limits of detection from thin films. The lowest values on soils are for malathion (1000 ppm range), and the largest for norflurazon (20,000 ppm), which is close to the limit of detection (LOD) found for the pesticide on the aluminum substrate (approximately 0.2 microg . cm(-2)). Different behaviors were observed as a function of time of storage in the ambient atmosphere or under vacuum; norflurazon adsorbed on soil exhibited high stability for a long period of time, and a rapid degradation of malathion with the elapsed time was clearly observed. The behavior of oxyfluorfen was also investigated but segregation processes seem to occur after several days. Although by far less sensitive than conventional methods based on extraction processes and used for real-world analytical applications, this technique is well suited to the study of the transformations occurring at the sample surface. A discussion is presented of the future prospects of such experiments in degradation studies. PMID- 16047318 TI - Comprehensive metabolic profiling of mono- and polyglutamated folates and their precursors in plant and animal tissue using liquid chromatography/negative ion electrospray ionisation tandem mass spectrometry. AB - This work reports the use of reversed-phase ion-pair chromatography coupled to electrospray ionisation mass spectrometry for the simultaneous profiling of folate-based metabolites including natural folates, their polyglutamatyl derivatives and their biosynthetic precursors in plant and animal tissue. A simple sample preparation method, using 0.1% citric acid and ascorbic acid in ice cold methanol, was used to extract and stabilise the folates, and three internal standards were used. Chromatography was on a C18 column using slow gradient elution with a mobile phase consisting of methanol/water with 5 mM dimethylhexylamine. Mass spectrometric detection was performed by multiple reaction monitoring in seven separate time windows in negative ion mode over the 25 min run time. Full, quantitative analysis was obtained for 16 folates and a 'semi-quantitative' analysis was possible for all other folates with up to eight conjugated glutamate residues by reference to structurally related calibration standards. The precision, accuracy and recovery of the method were generally within the accepted guidelines for a quantitative bioanalytical method and the method was linear over the range 0.2 to 10 ng of individual folate per sample. The method was applied to profile mono- and polyglutamated tetrahydrofolates (including subcellular analysis) in a range of plant species, including Arabidopsis, spinach, Brassica and wheat; the technique was also successfully applied to the profiling of folates in mouse tissue. PMID- 16047319 TI - Atmospheric O2, CO2 and delta13C measurements from aircraft sampling over Griffin Forest, Perthshire, UK. AB - Regular vertical aircraft sampling has been performed in the lower troposphere above Griffin Forest, near Aberfeldy, Perthshire, UK (56 degrees 37'N, 3 degrees 47'W), between February 2003 and May 2004, for analysis of O2/N2, CO2 and delta13C of CO2. We sampled flasks between 800 and 3100 m above sea level. The peak-to-peak amplitude of the seasonal cycle of O2/N2 decreases from 171 per meg at 800 m to 113 per meg at 3100 m. Furthermore, the seasonal cycle is shifted from low to high altitudes with a lag of about 1 month. The same features are observed for CO2 with a decrease in the peak-to-peak amplitude of the seasonal cycle from 17.6 ppm at 800 m to 11.4 ppm at 3100 m. The vertical profiles show decreasing O2/N2 ratios in summer and increasing O2/N2 ratios in wintertime with increasing sampling height, due to surface exchange of oxygen with the land biosphere and the ocean. The O2:CO2 exchange ratios of the vertical profiles vary between -1.5 and -2.4 mol O2/mol CO2. PMID- 16047320 TI - Buffer solution can control the porosity of DNA-chitosan complexes. AB - The current studies examine the pore properties and biological effects of DNA chitosan complexes, which may be useful as scaffolds for tissue engineering. The porosity of the DNA-chitosan complexes was controlled by rinsing them with several different pH 7.2 buffer solutions, including phosphate-buffered saline (PBS), Tris-HCl, boric acid, and N-(2-hydroxyethyl)piperazine-N'-(2-ethanesufonic acid) (HEPES). Rinsing with PBS resulted in 84% porosity, whereas rinsing with Tris-HCl produced 94% porosity. It was further found that daunorubicin hydrochloride complex intercalated with and bound to the groove of the DNA chitosan complexes, indicating that DNA in the complexes maintains its double stranded helical structure. The DNA-chitosan complexes were not toxic to MG-63 osteoblast-like cells and caused only a mild tissue response when implanted subcutaneously in the backs of rats. These results suggest that buffer-rinsed DNA chitosan complexes may be useful as a scaffold material in tissue engineering. PMID- 16047321 TI - Trophic level and macronutrient shift effects associated with the weaning process in the Postclassic Maya. AB - The weaning process was investigated at two Maya sites dominated by Postclassic remains: Marco Gonzalez (100 BC-AD 1350) and San Pedro (1400-AD 1650), Belize. Bone collagen and bioapatite were analyzed from 67 individuals (n < or = 6 years = 15, n > 6 years = 52). Five isotopic measures were used to reconstruct diet and weaning: stable nitrogen- and carbon-isotope ratios in collagen, stable carbon- and oxygen-isotope ratios in bioapatite, and the difference in stable carbon isotope values of coexisting collagen and bioapatite. Nitrogen-isotope ratios in infant collagen from both sites are distinct from adult females, indicating a trophic level effect. Collagen-to-bioapatite differences in infant bone from both sites are distinct from adult females, indicating a shift in macronutrients. Oxygen-isotope ratios in infant bioapatite from both sites are also distinct from adult females, indicating the consumption of breast milk. Among infants, carbon- and nitrogen-isotope ratios vary, indicating death during different stages in the weaning process. The ethnohistoric and paleopathological literature on the Maya indicate cessation of breast-feeding between ages 3-4 years. Isotopic data from Marco Gonzalez and San Pedro also indicate an average weaning age of 3-4 years. Based on various isotopic indicators, weaning likely began around age 12 months. This data set is not only important for understanding the weaning process during the Postclassic, but also demonstrates the use of collagen-to-bioapatite spacing as an indicator of macronutrient shifts associated with weaning. PMID- 16047322 TI - Chitosan/gelatin-based films crosslinked by proanthocyanidin. AB - Novel polymer networks consisting of crosslinked gelatin/chitosan were prepared by a solution casting technique. Methods for bulk crosslinking were developed to modify the gelatin/chitosan blends with the use of a nontoxic crosslinking reagent, proanthocyanidin (PA). FTIR spectral analyses of the preparations showed network formations of crosslinked gelatin, chitosan, and PA by amide and ester linkages. The crosslinked networks were stable in the aqueous state, and had improved mechanical properties and thermal stability when compared with nonlinked gelatin (G) and chitosan/gelatin (C/G) films. In vitro protease digestion and cell-culture studies showed that the PA-crosslinked C/G films are nontoxic and exhibited decreased biodegradation rate and a better ability to support cell adhesion and proliferation than noncrosslinked gelatin or chitosan alone. These results suggest that such a nontoxic crosslinked gelatin/chitosan scaffold can become a promising matrix for tissue-engineering applications. PMID- 16047323 TI - Mechanical evaluation of bone samples following alendronate therapy in healthy male dogs. AB - Alendronate and other bisphosphonates are clinically efficacious in treating postmenopausal osteoporosis, Paget's disease and hypercalcemia associated with malignancy. Because bisphosphonates are being considered for use in younger patients with joint replacements to prevent osteolysis, and for stress fracture prophylaxis in military recruits, it is important to know how bisphosphonate therapy affects healthy bone. We sought to determine whether bones from healthy male dogs exhibit alterations in structural or mechanical properties following alendronate treatment for 23 weeks. We tested trabecular tissue samples in compression and determined tissue ash density. We tested whole long bones in bending and torsion. For trabecular samples, we evaluated trabecular modulus, strength, and density. For whole bone specimens, we compared structural stiffness and ultimate load. We found no significant differences in any measure, between canines treated with alendronate for 23 weeks and controls, although we found consistent trends toward higher properties in the treated group. Correlation analysis revealed significant relationships between stiffness and strength measures for each mechanical test. Our results indicate bisphosphonate treatment in healthy canines does not weaken the properties of bone. The trends indicate a slight positive overall effect of alendronate treatment on the mechanical properties of healthy canine bone. PMID- 16047324 TI - Shipwrecks and founder effects: divergent demographic histories reflected in Caribbean mtDNA. AB - During the period of the Atlantic slave trade (15th-19th centuries), millions of people were forced to move from Africa to many American destinations, changing dramatically the human landscape of the Americas. Here, we analyze mitochondrial DNA from two different American populations with African ancestry, with hitherto unknown European and Native American components. On the basis of historical records, African-Americans from Choco (Colombia) and the Garifunas (or "Black Carib") of Honduras are likely to have had very different demographic histories, with a significant founder effect in the formation of the latter. Both the common features and differences are reflected in their mtDNA composition. Both show a minor component (approximately 16%) from Native Central/South Americans and a larger component (approximately 84%) from sub-Saharan Africans. The latter component is very diverse in the African-Americans from Choco, similar to that of sub-Saharan Africans, but much less so in the Garifunas, with several mtDNA types elevated to high frequency, suggesting the action of genetic drift. PMID- 16047325 TI - Sol-gel synthesis of bioactive glass scaffolds for tissue engineering: effect of surfactant type and concentration. AB - Well-defined structural characteristics are some of the exigencies that have to be attended when scaffolds for bone tissue cell culture are designed. A high porosity (70-90%) and a high specific surface area and an average pore size>150 microm will contribute to allow cell migration throughout the structure, adhesion, and proliferation. At the same time, the biodegradation of the material should occur in a proper rate. One way to reach a structure with these characteristics is to produce foams during sol-gel processing of bioactive glasses (system CaO--SiO2--P2O5). The addition of a surfactant in the sol-gel solution is necessary for foam formation and to maintain its stability until complete gelation occurs. This study presents the performance evaluation of two surfactants [sodium lauryl ether sulfate (SLES) and Tergitol] to determine optimum conditions for foaming ability and stability properties. The anionic surfactant SLES showed better results in terms of foam volume and its stability. Bioactive glass foams obtained with use of this surfactant presented a higher and interconnected porosity. The porosity of the scaffolds produced was 90%, and the macropore size ranged from 100 to 500 microm. PMID- 16047326 TI - The effect of curing modes on polymerization contraction stress of a dual cured composite. AB - Although a lower curing rate is often cited as the reason why a chemical cured (CC) dental composite produces lower polymerization contraction stress (PCS) than a light cured (LC) composite, the exact mechanism is still unclear. In addition, the comparison is often made by using different brands of composites. The comparison's fairness is questionable because the two composites have different compositions and preparation procedures. The goal of the present work was to determine if the curing mode alone can produce different PCS. We formulated a dual cured composite and prepared it the same way for both CC and LC modes. We measured PCS by a strain gauge method, shrinkage by a video-imagining technique, degree of conversion (DC) by infrared spectroscopy, and flexural modulus by the three-point bending test. The CC specimens showed lower PCS and lower flexural modulus than the LC specimens, although both possessed an identical chemical composition and physical texture before cure. This finding indicates that the curing mode alone can affect PCS. Because the CC and LC specimens produced a similar shrinkage and DC, the lower modulus is considered to be one of the reasons for the lower stress. Using a structural inhomogeneity model, we explained how a resin composite with an identical DC can have different physical properties such as the modulus. PMID- 16047327 TI - Accelerating perfusion process optimization by scanning non-steady-state responses. AB - Perfusion processes provide consistent culture conditions, high productivity and low product residence times. However, process development can be slow due to the 1 week or more required to reach each steady state. The objective of this work was to accelerate process development in perfusion cultures by scanning non steady-state transient responses to qualitatively predict steady-state performance. The method was tested using a shift in temperature every 3 days, scanned down by steps of 2 degrees C from 37 degrees C to 31 degrees C, then scanned up to 37 degrees C. Higher t-PA concentrations were predicted at lower temperatures, confirmed by subsequent pseudo-steady-state results. In most cases, transient values on the 3rd day were in close concordance with pseudo-steady state values. To further accelerate process development, transient scanning was applied to small-scale, non-instrumented cultures. Similar results were obtained, although quantitative t-PA values were 15-30 times lower than in high cell density perfusion cultures. The method was further explored by investigating 1 day transient shifts in temperature where more variability was observed, suggesting that the cells were still adapting to the new environment. Nonetheless, the overall response again qualitatively predicted the pseudo-steady state temperature response. Use of transient scanning in conjunction with pseudo steady-state verification and refinement of optimal results could reduce process development time to a third or less of comparable steady-state-based optimization. PMID- 16047328 TI - Bone formation using human adipose tissue-derived stromal cells and a biodegradable scaffold. AB - Human adipose tissue, obtained by liposuction, was processed to obtain a fibroblast-like population of cells or adipose tissue-derived stromal cells (ATSCs). The ATSCs, as well as bone marrow-derived mesenchymal stem cells (BMSCs), have the capacity for renewal and the potential to differentiate into multiple lineages of mesenchymal tissues. These cells are capable of forming bone when implanted ectopically in an appropriate scaffold. The aim of this study was to evaluate a beta-tricalcium phosphate (beta-TCP) as a scaffold and to compare the potential of osteogenic differentiation of ATSCs with BMSCs. Both cell types were loaded into beta-TCP disk and cultured in an osteogenic induction medium. Optimal osteogenic differentiation in ATSCs in vitro, as determined by secretion of osteocalcin, scanning electron microscope, and histology, were obtained in the culturing with the beta-TCP disk. Furthermore, bone formation in vivo was examined by using the ATSC- or BMSC-loaded scaffolds in nude mice. The present results show that ATSCs have a similar ability to differentiate into osteoblasts and to synthesize bone in beta-TCP disk as have BMSCs. PMID- 16047329 TI - Near infrared laser-tissue welding using nanoshells as an exogenous absorber. AB - BACKGROUND AND OBJECTIVE: Gold nanoshells are a new class of nanoparticles that can be designed to strongly absorb light in the near infrared (NIR). These particles provide much larger absorption cross-sections and efficiency than can be achieved with currently used chemical chromophores without photobleaching. In these studies, we have investigated the use of gold nanoshells as exogenous NIR absorbers to facilitate NIR laser-tissue welding. STUDY DESIGN/MATERIALS AND METHODS: Gold nanoshells with peak extinction matching the NIR wavelength of the laser being used were manufactured and suspended in an albumin solder. Optimization work was performed on ex vivo muscle samples and then translated into testing in an in vivo rat skin wound-healing model. Mechanical testing of the muscle samples was immediately performed and compared to intact tissue mechanical properties. In the in vivo study, full thickness incisions in the dorsal skin of rats were welded, and samples of skin were excised at 0, 5, 10, 21, and 32 days for analysis of strength and wound healing response. RESULTS: Mechanical testing of nanoshell-solder welds in muscle revealed successful fusion of tissues with tensile strengths of the weld site equal to the uncut tissue. No welding was accomplished with this light source when using solder formulations without nanoshells. Mechanical testing of the skin wounds showed sufficient strength for closure and strength increased over time. Histological examination showed good wound-healing response in the soldered skin. CONCLUSIONS: The use of nanoshells as an exogenous absorber allows the usage of light sources that are minimally absorbed by tissue components, thereby, minimizing damage to surrounding tissue and allowing welding of thicker tissues. PMID- 16047330 TI - MammoSite excision volume as a predictor for residual disease. AB - BACKGROUND: The MammoSite catheter is a brachytherapy device used for accelerated partial breast irradiation. Currently, it is available as 2 spherically shaped balloons meant to fill 70-cc and 120-cc cavity volumes. This study was designed to define the relation of these excision volumes to the likelihood of microscopically detectable, residual disease based on tumor size, margin status, patient age, and histology. METHODS: The study data base was comprised of 531 patients with Stage 0, I, and II breast carcinoma (using American Joint Committee on Cancer staging criteria) who received breast-conserving therapy and underwent surgical reexcision. Patients in the data base were stratified based on the volume of their initial excision: < or = 70 cc versus > 70 cc and < or = 120 cc versus > 120 cc. RESULTS: Surgical margin size was found to be a strong predictor of residual disease both for patients with smaller excision volumes (P = 0.0014) and patients with larger excision volumes (P = 0.0003); histology (extensive intraductal component [EIC] or pure ductal carcinoma in situ [DCIS]) also was a strong predictor. Tumor size was significant only for the larger volume group (P = 0.029). On multivariate analysis, only histology and initial margin status were significant correlates with residual disease. The adjusted odds ratio for residual disease with pure DCIS was 0.79, and the adjusted odds ratio for invasive ductal or lobular carcinoma (IDC/ILC) without EIC was 0.44 relative to IDC/ILC with EIC (P = 0.008). The adjusted odds ratio for residual disease with a positive initial margin versus a negative initial margin was 2.65 (P < or = 0.0001). CONCLUSIONS: For excisions amenable to use of the MammoSite catheter, a margin > or = 1.0 mm appeared to afford at most a 35% risk of microscopically detectable residual tumor. Evidence of EIC on excision of IDC/ILC connoted a significantly higher risk. Age did not appear to be predictive for residual disease. PMID- 16047331 TI - Evaluation of breast cancer with a computer-aided detection system by mammographic appearance and histopathology. AB - BACKGROUND: The objective of this study was to evaluate the performance of a computer-aided detection (CAD) system for the detection of breast cancer, based on mammographic appearance and histopathology. METHODS: From 1000 consecutive screening mammograms from women with biopsy-proven breast carcinoma, 273 mammograms were selected randomly for retrospective evaluation by CAD. The sensitivity of the CAD system for breast cancer was assessed from the proportion of masses and microcalcifications detected. The corresponding tumor histopathologies also were evaluated. Normal mammograms (n = 155 patients) were used to determine the false-positive rate of the system. RESULTS: Of the 273 breast carcinomas, 149 appeared mammographically as masses, and 88 appeared as microcalcifications, including 36 carcinomas that presented as mixed lesions. The CAD system marked 125 of 149 masses correctly (84%), marked 86 of 88 microcalcifications correctly (98%), and marked 32 of 36 of mixed lesions correctly (89%.). The system showed a high sensitivity for the detection of ductal carcinoma in situ (95%; 73 of 77 lesions), invasive lobular carcinoma (95%; 18 of 19 lesions), invasive ductal carcinoma (85%; 125 of 147 lesions), and invasive mammary carcinoma (90%; 27 of 30 lesions). The highest CAD system sensitivity was for all invasive carcinomas that presented as microcalcifications (100%). On normal mammograms, there was an average of 1.3 false-positive CAD marks per image. CONCLUSIONS: The CAD system correctly marked a large majority of biopsy-proven breast cancers, with a greater sensitivity for lesions with microcalcifications and without significant impact of performance based on tumor histopathology. CAD was highly effective in detecting invasive lobular carcinoma (sensitivity, 95%) and ductal carcinoma in situ (sensitivity, 95%). CAD represents a useful tool for the detection of breast cancer. PMID- 16047332 TI - Quality of prostate carcinoma care in a statewide public assistance program. AB - BACKGROUND: The authors evaluated the feasibility of measuring quality of care in a statewide public assistance program for men with prostate carcinoma. METHODS: The sample consisted of 84 men who were followed for > or = 6 months after receiving primary treatment for early-stage prostate carcinoma (55 received radical prostatectomy and 29 received radiotherapy) through a free public program for low-income, uninsured men. Quality was assessed by chart review with 16 indicators previously developed and validated at the RAND Corporation, as well as by telephone and mail surveys that included the University of California at Los Angeles Prostate Cancer Index short form. RESULTS: Quality of care measurement was feasible for 13 (81%) indicators from electronic chart abstraction, administrative documents, and patient questionnaires. Communication between specialist and primary physician was better for men treated with radiotherapy than with surgery (84% vs. 45%, P = 0.004). Subjects treated in private institutions were more likely than those treated in public institutions to have > or = 2 follow-up visits with the treating physician or institution within 1 year of treatment (93% vs. 63%, P = 0.003) and to have documentation of communication with the primary care physician (90% vs. 40%, P << 0.0001). Disease-specific, health-related quality of life 6 months after treatment did not appear to differ between public and private facilities. CONCLUSIONS: The authors found the application of quality of care indicators to be feasible in a statewide public assistance program, but with some differences between public and private providers. These quality of care indicators identified target areas for improvement. PMID- 16047333 TI - Prognostic impact of BRCA1 pathogenic and BRCA1/BRCA2 unclassified variant mutations in patients with ovarian carcinoma. AB - BACKGROUND: The clinical relevance of BRCA1/2 alterations in ovarian carcinoma patients is debatable. Our aim was to determine factors influencing the risk of recurrence and death in ovarian carcinoma patients with BRCA pathogenic and unclassified variant mutations. METHODS: A consecutive series of 205 women with primary ovarian carcinoma were screened for mutations in BRCA1 and BRCA2 genes using a conformational sensitive gel electrophoresis and direct sequencing. Data regarding medical and familial history were collected using questionnaires. Clinical and pathologic data were extracted from medical records. RESULTS: Unclassified variant mutations in BRCA1 or BRCA2 genes were found in 16 (8%) patients, and BRCA1 pathogenic mutations were found in 18 (9%) patients. No pathogenic mutation was found in BRCA2 gene. Multivariate analysis showed that BRCA1 pathogenic mutation was an independent predictor of reduced risk of relapse and death (Hazard ratios [HR] 0.52 [confidence interval {CI} 0.28-0.98] and 0.38 [CI 0.10-0.96], respectively). Unclassified variant mutation did not affect recurrence and survival (HR 0.84 [CI 0.43-1.66] and 0.94 [CI 0.48-1.82], respectively). Other factors associated with reduced risk of relapse and death were complete pathologic remission at second-look laparotomy and family history of breast and ovarian carcinoma, respectively. Recurrence and death outcomes among unclassified variant mutation carriers did not differ significantly from those in sporadic cases. CONCLUSIONS: Patients with BRCA1 pathogenic mutation seem to have reduced risk of recurrence and death. These results should be interpreted with caution as they may be influenced by more intensive treatment, better response to cisplatin, and younger age of mutation carriers. Clinical relevance of BRCA1/2 unclassified variant mutations warrants further studies. PMID- 16047334 TI - Leukemic transformation of polycythemia vera: a single center study of 23 patients. AB - BACKGROUND: Acute leukemia (AL) may occur as rare and late event of polycythemia vera (PV). METHODS: The current study included 23 patients who developed acute leukemia in a cohort of 414 consecutive PV patients with long-term observation (3208 person years of follow-up). Kaplan-Meier Product-Limit method was used to estimate the cumulative probability of survival; Gehan-Wilcoxon test was applied to compare survival in different groups of patients. RESULTS: Median age was 68 years, and 18 patients (78%) were > 60 years of age. At diagnosis of AL, most patients had a white blood count > 10 x 10(9)/L (n = 17; 74%), Hgb < 10 g/dL (n = 13; 57%), and platelet count > 50 x 10(9)/L (n = 17; 74%). Of 14 patients in whom cytogenetic analysis was available at leukemic transformation, 12 showed high risk abnormalities including complex karyotype (n = 10), del (7)(q22) sole (n = 1) and del (X)(q26) sole (n = 1), whereas 2 had a normal karyotype. In patients whose karyotype was available at diagnosis of PV, cytogenetic evolution was documented at progression to AL. Treatment consisted of supportive care and/or low-dose chemotherapy (n = 15), or induction chemotherapy (n = 8). This included idarubicin plus cytarabine (n = 3), high-dose cytarabine (n = 4), and fludarabine based regimen (n = 1). Allogenic stem cell transplantation was offered to a single patient, who is alive at Day + 70. The outcome of patients was poor, with a median survival of 2.9 months (range, 0.6-20.1 mos), with no significant differences between palliation and intensive treatments. CONCLUSIONS: AL following PV has distinct clinical and biologic features. Outcome of patients is poor irrespective of the treatment employed. PMID- 16047335 TI - A metaanalysis of 18F-2-deoxy-2-fluoro-D-glucose positron emission tomography in the staging and restaging of patients with lymphoma. AB - BACKGROUND: In recent years, the use of positron emission tomography (PET) has become widespread for the staging and follow-up of several malignancies. In the current study, the authors conducted a metaanalysis of the published literature to evaluate the diagnostic performance of 18F-2-deoxy-2-fluoro-D-glucose PET (FDG PET) in the staging of patients with lymphoma. METHODS: The authors conducted a systematic MEDLINE search of articles published between January 1995 and June 2004. Studies that evaluated FDG-PET with a dedicated camera and that reported sufficient data to permit the calculation of sensitivity and specificity were included in the analysis. Two reviewers independently reviewed the eligibility of the studies and abstracted data (sample population; characteristics of FDG-PET; and the number of true-positive results, true-negative results, false-positive results, and false-negative results). The authors estimated the pooled sensitivity, false-positive rate, and maximum joint sensitivity and specificity. RESULTS: Twenty studies were eligible for the metaanalysis. Fourteen studies included patient-based data, comprising a sample size of 854 subjects, and 7 studies included lesion-based data, totaling 3658 lesions. Among those studies with patient-based data, the median sensitivity was 90.3% and the median specificity was 91.1%. The pooled sensitivity was 90.9% (95% confidence interval [95% CI], 88.0-93.4) and the pooled false-positive rate was 10.3% (95% CI, 7.4 13.8). The maximum joint sensitivity and specificity was 87.8% (95% CI, 85.0 90.7). The pooled sensitivity and false-positive rate appeared to be higher in patients with Hodgkin disease compared with those with non-Hodgkin lymphoma. CONCLUSIONS: The results of the current study indicate that FDG-PET is a valuable tool for the staging and restaging of patients with lymphoma; showing a high positivity and specifity. Clinicians may consider adding FDG-PET to the staging workup of patients with lymphoma. PMID- 16047336 TI - Mating of Xenos vesparum (Rossi) (Strepsiptera, Insecta) revisited. AB - The controversial mating of the strepsipteran Xenos vesparum was studied to investigate the possible sperm routes for fertilization. The female, which is a neotenic permanent endoparasite of Polistes wasps, extrudes only its anterior region, the "cephalothorax," from the host abdomen. This region has an opening where both mating and larval escape occur. Observations with scanning and transmission electron microscopy revealed spermatozoa not only in the hemocoel, but also in the "ventral canal" (an extragenital duct peculiar to strepsipteran females) and in the "genital ducts" (ectodermal invaginations connecting the ventral canal to the hemocoel) of recently mated females. Xenos vesparum spermatozoa can reach the oocytes either through the hemocoel as a result of a hypodermic insemination, or by moving along the extragenital ducts, which are later used by first instar larvae to escape. The hypothesis of hypodermic insemination is reconsidered in the light of behavioral and ultrastructural evidence. PMID- 16047337 TI - CD72-mediated suppression of human naive B cell differentiation by down regulating X-box binding protein 1. AB - B cells can differentiate into antibody-secreting plasma cells, however the signals that control the entry into this pathway are not clearly understood. We have investigated the role of human CD72 in mature B cell differentiation. Human CD72 is preferentially expressed in naive B cells, but marginal levels of expression can be found in switched memory B cells. CD72 cross-linking promoted an increase in B cell activation and proliferation. Interestingly, expression of CD27, whose signal induces the differentiation of B cells into plasma cells, was down-modulated by CD72 stimulation. This CD72 signaling also induced tyrosine phosphorylation of various proteins such as Blk. Plasma cell differentiation and Ig syntheses were diminished by CD72 ligation in the presence of Staphylococcus aureus Cowan strain (SAC) plus IL-2 but not in the presence of CD40 signaling or CpG oligodeoxynucleotide. Our results show that CD72 signaling reduces the expression of X-box binding protein 1 in B cells stimulated with SAC plus IL-2, but the expression of PRDI-BF1 was unaffected. Taken together, these data demonstrate that CD72 is a key molecule in regulating mature B cell differentiation, particularly in preventing the differentiation of naive B cells into plasma cells, thus blocking the production of low-affinity antibodies. PMID- 16047338 TI - CpG-activated Thy1.2+ dendritic cells protect against lethal Listeria monocytogenes infection. AB - Synthetic oligodeoxynucleotides containing CpG motifs (CpG ODN) activate the innate immune system by interacting with Toll-like receptor 9. The resultant immune response increases host resistance to infection by a variety of pathogenic microorganisms, including Listeria monocytogenes. There is a considerable interest in harnessing the immunoprotective properties of CpG ODN, yet little is known of the cell phenotype(s) responsible for mediating this protection. This work demonstrates that treatment of mice with CpG ODN increases the number of Thy1.2+, CD11c+ dendritic cells (Thy1.2+ DC) in the spleen, which are both necessary and sufficient for transferring resistance to infection from CpG treated donors to naive recipients. These CpG-activated Thy1.2+ DC are distinct from conventional (CD11c(hi), Thy1.2-) or plasmacytoid DC (mPDCA+), and secrete IFN-gamma that contributes to protection. These findings suggest that a novel Thy1.2+ DC subset plays a critical role in mediating the immunoprotective activity of CpG DNA. PMID- 16047339 TI - Induction of thymocyte apoptosis by systemic administration of concanavalin A in mice: role of TNF-alpha, IFN-gamma and glucocorticoids. AB - Administration of concanavalin A (Con A) is a well-established model of acute immune-mediated hepatitis. Here, we demonstrate that intravenous injection of Con A in mice induces profound thymic atrophy. Compared to liver damage, the kinetics of Con A-induced thymic atrophy is slower and more prolonged; the nadir in thymocyte number is reached 4 days after Con A injection, whereas peak transaminase levels are observed at 12-24 h. Marked alterations in the ratio of CD4+ and CD8+cells in the thymus and spleen and significantly increased rates of thymocyte and splenocyte apoptosis are observed. Neutralization of the cytokines TNF-alpha or IFN-gamma, which protects mice from Con A-induced hepatitis, prevents thymic atrophy as well as alterations in CD4+ and CD8+ cell numbers and apoptosis rates. However, neither TNF-alpha nor IFN-gamma are detectable in thymocyte lysates after Con A injection, whereas both cytokines are present in liver, spleen and serum. Administration of the glucocorticoid receptor antagonist mifepristone does not prevent thymic atrophy, thus ruling out a possible contribution of endogenous glucocorticoids. Con A-induced thymic atrophy is accompanied by down-regulation of Bcl-2 expression in the thymus, which is prevented by neutralization of TNF-alpha or IFN-gamma. These data demonstrate that the thymus is a critical target organ of Con A-induced inflammation; the effects of Con A on the thymus are mediated by extrathymic production of TNF alpha and IFN-gamma, but not by glucocorticoids. PMID- 16047340 TI - IL-15 induces mast cell migration via a pertussis toxin-sensitive receptor. AB - IL-15 induces proliferation, inhibits apoptosis and increases IL-4 production in murine mast cells. There is evidence that these activities are mediated via the uncharacterised receptor, IL-15R-X, rather than the classical three-chain IL-15 receptor. Effects of IL-15 on important aspects of mast cell biology, such as migration and degranulation, are unknown. We report that IL-15 induces migration of murine and human mast cells in a dose-dependent and biphasic manner, with peaks of migration occurring at approximately 10(-15) and approximately 10(-9) M. The potency of the response was similar to that induced by other well-established mast cell chemoattractants. Competition assays performed with murine and human mast cells indicate that both peaks of migration are due to chemotaxis. Pre treatment of cells with pertussis toxin (PTX), a guanine nucleotide-binding regulatory protein (G-protein) inhibitor, resulted in complete inhibition of murine mast cell migration at approximately 10(-15) M IL-15, and human mast cell migration at approximately 10(-15) and approximately 10(-9) M. This demonstrates that murine and human mast cells express a PTX-sensitive receptor, activated in response to IL-15. Additionally, IL-15 did not induce degranulation in murine mast cells. Locally-produced IL-15 may contribute to mast cell recruitment during inflammatory responses, thereby acting as a linking cytokine between innate and adaptive arms of the immune system. PMID- 16047341 TI - Evidence for an enhanced adhesion of DC to fibronectin and a role of CCL19 and CCL21 in the accumulation of DC around the pre-diabetic islets in NOD mice. AB - The non-obese diabetic (NOD) mouse is a widely used animal model for the study of human diabetes. The lymphocytic (peri-)insulitis is preceded by an early accumulation of dendritic cells (DC) around the islets of Langerhans. This DC accumulation is thought to derive from an influx of monocytes attracted by pro inflammatory chemokines. Besides chemokines, extracellular matrix (ECM) proteins play an important role in the accumulation of leukocytes in tissues. We studied the expression of the chemokines CCL2, CCL5, CXCL10, CCL19 and CCL21 over time in pancreases of NOD and control mice by ELISA on pancreas lysates as well as by immunohistochemistry. In addition, we studied the adhesive capacity of bone marrow-derived DC (BMDC) to ECM components. DC in the NOD pancreas accumulated at sites with an intense expression of fibronectin. In vitro, NOD BMDC showed increased fibronectin adhesion and increased VLA-5 expression. At the time of early DC accumulation (<10 wk), the lymphoid tissue-related chemokines CCL19 and CCL21 were increased. Our findings support the view that the early accumulation of DC around the NOD islets is not the consequence of an enhanced attraction of precursors and immature DC by pro-inflammatory chemokines. It rather might be the consequence of an aberrantly enhanced adhesion and retention of NOD DC. PMID- 16047342 TI - Notch ligands Delta-like1, Delta-like4 and Jagged1 differentially regulate activation of peripheral T helper cells. AB - The Notch pathway is involved in cell differentiation processes in various organs and at several developmental stages. The importance of Notch for early T lymphocyte development is well established. Recently, Notch has been implicated in directing naive T helper cell differentiation towards the Th1, Th2 or regulatory T cell lineages. However, the molecular events underlying these processes are poorly understood. We show that the Notch ligands Delta-like1, Delta-like4 and Jagged1 differentially affect early T cell activation and proliferation following T cell receptor cross-linking. Delta-like1 and Jagged1 induce a dose-dependent inhibition of early activation markers CD69 and CD25, as well as inhibition of proliferation after anti-CD3 stimulation of purified CD4+ T cells. Similarly, the rapid activation of transcription factors NF-AT, AP-1 and NF-kappaB is suppressed. In contrast, triggering of Notch by Delta-like4 enhances T cell activation and proliferation. The observed effects are dependent on simultaneous cross-linking of TCR and Notch but independent of gamma-secretase mediated cleavage of Notch. These data suggest direct interference between Notch and early TCR signal transduction events, independent of the classical Notch pathway via release of the Notch intracellular domain. A Notch-mediated alteration of TCR signaling strength may contribute to the recently described modulation of naive T cell differentiation by Notch ligands. PMID- 16047343 TI - A phase I study and pharmacologic evaluation of irinotecan and carboplatin for patients with advanced ovarian carcinoma who previously received platinum containing chemotherapy. AB - BACKGROUND: The objectives of the current study were to determine the maximum tolerated dose (MTD) of irinotecan and carboplatin in combination, to evaluate the efficacy and toxicity of the combination in patients with advanced ovarian carcinoma who previously received platinum-containing chemotherapy, and to examine the pharmacokinetics and pharmacodynamics of both drugs by using the Chatelut formula. METHODS: Patients with advanced ovarian carcinoma who previously received platinum-containing chemotherapy were treated with a combination of irinotecan and carboplatin. Carboplatin was administered as a 60 minute intravenous infusion on Day 1 and was followed by irinotecan, which was administered as a 90-minute intravenous infusion on Days 1, 8, and 15. Six dose levels of irinotecan (in mg/m(2))/carboplatin (mg . mL/min) were planned: 50 mg/m(2)/4 mg . mL/minute, 60 mg/m(2)/4 mg . mL/minute, 50 mg/m(2)/5 mg . mL/minute, 60 mg/m(2)/5 mg . mL/minute, 50 mg/m(2)/6 mg . mL/minute, and 60 mg/m(2)/6 mg . mL/minute. The carboplatin dosage was calculated by using the Chatelut formula. Treatment was repeated at 28-day intervals. RESULTS: In total, 19 patients in cohorts of 3 to 5 patients received irinotecan and carboplatin at 5 dose levels. The dose-limiting toxicities were Grade 4 neutropenia and Grade 4 thrombocytopenia. The MTD of the irinotecan/carboplatin combination was 60 mg/m(2)/5 . mg mL/minute. Partial responses were observed at higher dose levels. Pharmacologic studies demonstrated that administration of the dosage estimated with the Chatelut formula instead of the Chatelut formula with adjustment for serum creatinine resulted in a slightly excessive dose of carboplatin. CONCLUSIONS: The recommended dose for the Phase II study was irinotecan 60 mg/m(2) on Days 1, 8, and 15 with carboplatin 5 mg/mL . minute on Day 1 repeated every 4 weeks. PMID- 16047344 TI - Spectrum and prevalence of BRCA1 and BRCA2 germline mutations in Sardinian patients with breast carcinoma through hospital-based screening. AB - BACKGROUND: Factors that are predictive of carrying BRCA1 and BRCA2 germline mutations in patients with breast carcinoma are awaited widely. The genetically homogeneous Sardinian population may be useful for defining the role of such genetic alterations further through a clinical evaluation program. METHODS: One hundred two of 659 patients with breast carcinoma (15.5%) who were collected consecutively had a family history of breast carcinoma and were screened for BRCA1/2 mutations by denaturing high-performance liquid chromatography and DNA sequencing. RESULTS: Three deleterious germline BRCA1/2 mutations were detected in 15 of 102 families (14.7%), including 13 families (86.7%) with BRCA2 mutations and 2 families (13.3%) with BRCA1 mutations. A single variant, BRCA2-8765delAG, was the most recurrent mutation in the series and was found in 12 of 102 families (11.8%) and in 18 of 657 patients (2.7%). The average age at diagnosis was significantly younger in families with BRCA1/2 mutations (48.6 yrs) compared with the age of patients who had no detectable mutation (52.9 yrs; P = 0.039). Moreover, BRCA1/2 mutations were found at a significantly higher rate in families who had at least 1 member with ovarian carcinoma or male breast carcinoma (5 of 12 families; 41.7%) than in families without such an association (10 of 90 families; 11.1%; P = 0.003). CONCLUSIONS: BRCA2 mutations were approximately 6 times more prevalent than BRCA1 mutations. A diagnosis of breast carcinoma before age 50 years, ovarian carcinoma, male breast carcinoma, and 3 affected generations all were associated significantly with BRCA1/2 mutations. Although the current findings provided further support for the hypothesis that additional breast carcinoma susceptibility genes remain to be identified, such indicators of the presence of BRCA1/2 mutations may be useful in counseling patients about undergoing genetic testing. PMID- 16047345 TI - Prognostic significance of the lymphocyte-to-neutrophil ratio in percutaneous fine-needle aspiration biopsy specimens of advanced nonsmall cell lung carcinoma. AB - BACKGROUND: The prognostic significance of tumor-infiltrating lymphocytes (TILs) in surgically resected carcinomas was reported. To apply this to inoperable nonsmall cell lung carcinomas (NSCLC) of Stage IIIB-IV, the authors estimated the occurrence of TILs using percutaneous fine-needle aspiration biopsy specimens, and tested the validity of this method. METHODS: The authors defined the L-N index as [L(S)/(L(S) + N(S)) - L(B)/(L(B) + N(B))], in which L(S) and N(S) denoted lymphocyte and neutrophil counts in the aspiration smear, and L(B) and N(B) denoted lymphocyte and neutrophil counts in the peripheral blood specimen. The cutoff value was set at twice the standard deviation of the L-N index of 41 smears contaminated with abundant blood. Retrospectively, the authors compared the survival rate of the group with a high L-N index (lymphocyte-dominant group) (n = 12) with the survival rate of the group with a low L-N index (lymphocyte nondominant group) (n = 60). Then, they performed a prospective study and compared the survival rates of these 2 groups (n = 21 and n = 54). The Cox proportional hazards model was used to determine the effect of the L-N index as a continuous variable and other prognostic factors. The correlation (r) between the L-N index-based grouping (L-N grouping) and the histologic grade of TILs was studied among resected lung tumor specimens (n = 164). RESULTS: In the retrospective and prospective studies, the survival rate was significantly higher in the lymphocyte-dominant group than in the lymphocyte-nondominant group (P = 0.0019 and P = 0.0001). Using multivariate analysis, the L-N index was an independent prognostic factor. A significant correlation was noted between L-N grouping and histologic grade of TILs (r = 0.476). CONCLUSIONS: The L-N index of aspiration smears was found to be an independent prognostic factor for patients with advanced-stage NSCLC. L-N grouping was correlated with the histologic assessment of TILs. PMID- 16047346 TI - Prolonged severe pancytopenia preceding the cutaneous lesions of juvenile xanthogranuloma. AB - We report a case of juvenile xanthogranuloma (JXG) having progressive pancytopenia for 6 months until the proliferating skin lesions. A 2-month-old infant presented recurrent fever, anemia, and hepatosplenomegaly mimicking hemophagocytic lymphohistiocytosis (HLH) or juvenile myelomonocytic leukemia (JMML). At 8 months of age, the biopsy of a growing papule on the elbow made the diagnosis. Bone marrow (BM) specimens showed clustering foamy cells including hemophagocytosis by histiocytes. Treatment with etoposide followed by vinblastine plus prednisolone (PSL) therapy improved the disease. Although JXG is a benign non-Langerhans cell histiocytosis, the multisystem-visceral form should be considered as a potential aggressive disease when associated with BM failure in early infancy. PMID- 16047347 TI - Re: Incidental detection of neuroblastoma and "Wait and See" strategy. PMID- 16047348 TI - Symptomatic hypoglycemia: an unusual side effect of oral purine analogues for treatment of ALL. AB - Symptomatic hypoglycemia is an unusual complication in children receiving oral purine analogues for treatment of childhood acute lymphoblastic leukemia (ALL). The exact mechanism of the hypoglycemic effect of the antimetabolic therapy remains unclear. Reduced hepatic glycogen stores or impaired hepatic glyconeogenesis may partly explain the hypoglycemia. To prevent hypoglycemia, food containing complex carbohydrates is recommended before sleep. In severe cases of hypoglycemia due to 6-mercaptopurine (6-MP), the dose can be given in the morning and if this fails 6-MP can be discontinued for a short period of time. We report a 3-year-old child who developed severe early morning hypoglycemia episodes that resolved after decreasing 6-MP while receiving non high risk ALL therapy. PMID- 16047349 TI - Dominant form of vanishing white matter-like leukoencephalopathy. AB - Leukoencephalopathy with vanishing white matter syndrome (childhood ataxia with central nervous system hypomyelination/vanishing white matter disease) is an autosomal recessive disorder characterized by the occurrence of acute episodes of deterioration after minor head trauma or infection, and symmetrical demyelination on magnetic resonance with cavitation aspects. Mutations in each of the five subunits of eIF2B have been identified. We report in an affected man and his mother an adult-onset form of childhood ataxia with central nervous system hypomyelination/vanishing white matter disease-like disorder with no mutations in the EIF2B genes and normal guanine nucleotide exchange factor eIF2B activity, suggesting a new dominant inheritance of this syndrome that may involve other genes. PMID- 16047350 TI - Clonal divergence and genetic heterogeneity in clear cell renal cell carcinomas with sarcomatoid transformation. AB - BACKGROUND: Approximately 5% of clear cell renal cell carcinomas contain components with sarcomatoid differentiation. It has been suggested that the sarcomatoid elements arise from the clear cell tumors as a consequence of clonal expansions of neoplastic cells with progressively more genetic alterations. Analysis of the pattern of allelic loss and X-chromosome inactivation in both the clear cell and sarcomatoid components of the same tumor allows assessment of the genetic relationship of these tumor elements. METHODS: The authors of the current study examined the pattern of allelic loss in clear cell and sarcomatoid components of renal cell carcinomas from 22 patients who had tumors with both components. DNA samples were prepared from formalin-fixed, paraffin-embedded renal tissue sections using laser-capture microdissection. Five microsatellite polymorphic markers for putative tumor suppressor genes on 5 different chromosomes were analyzed. These included D3S1300 (3p14), D7S522 (7q31), D8S261 (8p21), D9S171 (9p21), and TP53 (17p13). In addition, X-chromosome inactivation analysis was performed in 14 tumors from female patients. RESULTS: The clear cell components showed loss of heterozygosity (LOH) at the D3S1300, D7S522, D8S261, D9S171, and TP53 loci in 18% (4/22), 18% (4/22), 50% (10/20), 15% (3/20), and 20% (4/20) of informative cases, respectively. LOH in the sarcomatoid components was seen at the D3S1300, D7S522, D8S261, D9S171, and TP53 loci in 18% (4/22), 41% (9/22), 70% (14/20), 35% (7/20), and 20% (4/20) of informative cases, respectively. Six cases demonstrated an LOH pattern in the clear cell component that was not seen in the sarcomatoid component. Different patterns of allelic loss were seen in the clear cell and sarcomatoid components in 15 cases. Clonality analysis showed the same pattern of nonrandom X-chromosome inactivation in both clear cell and sarcomatoid components in 13 of the 14 cases studied. One case showed a random pattern of X-chromosome inactivation. CONCLUSION: X chromosome inactivation analysis data suggest that both clear cell and sarcomatoid components of renal cell carcinomas are derived from the same progenitor cell. Different patterns of allelic loss in multiple chromosomal regions were observed in clear cell and sarcomatoid components from the same patient. This genetic heterogeneity indicates genetic divergence during the clonal evolution of renal cell carcinoma. PMID- 16047351 TI - Prognostic value of MYCN and ID2 overexpression in neuroblastoma. AB - BACKGROUND: The molecular mechanisms driven by overexpression of the MYCN gene in neuroblastoma are not well known. Whether MYCN overexpression in the absence of genomic amplification, or ID2 overexpression has prognostic value remains controversial. PROCEDURE: Ninety-nine neuroblastic tumors from Memorial Sloan Kettering Cancer Center and 12 neuroblastoma cell lines were analyzed by Affymetrix U95v2 Microarray System. The expression levels of the genes MYCN and ID2 were determined by RT-PCR and immunohistochemistry and the clinical value of the overexpression of both genes was determined. RESULTS: MYCN genomic amplification with overexpression was prognostic for survival (P = 0.0005). Stage 4 patients with MYCN amplification but without overexpression, had no increased likelihood of death, whereas cases with MYCN overexpression but no genomic amplification showed a low survival (P = 0.0096). ID2 did not correlate with MYCN expression (R = -0.23 for tumors and -0.27 for cell lines) or with survival (P = 0.8746). CONCLUSIONS: MYCN amplification was not related to clinical outcome in the absence of overexpression in neuroblastoma tumors. ID2 expression appears to be independent of MYCN expression and lacks prognostic value. PMID- 16047352 TI - Combined-modality treatment for isolated recurrences of breast carcinoma: update on 30 years of experience at the University of Texas M.D. Anderson Cancer Center and assessment of prognostic factors. AB - BACKGROUND: In three prospective, single-arm studies, the authors previously showed an improved outcome for anthracycline-naive patients with isolated sites of recurrent breast carcinoma (BC) who were treated with doxorubicin-based chemotherapy after local therapy (surgery and/or radiotherapy). In the current report, the initial results are presented from a Phase II trial of docetaxel (100 mg/m(2) every 21 days for 6 cycles) given after local therapy for recurrent BC (Stage IV BC with no evidence of clinically measurable disease) in patients who received prior adjuvant anthracycline-based chemotherapy, and the authors provide an update of the 3 previous studies. An analysis of prognostic factors for these patients also is presented. METHODS: Eligibility criteria for all studies included histologic proof of recurrent BC that had been resected and/or irradiated with curative intent. Survival was calculated using the Kaplan-Meier method. Univariate survival analyses were performed to test for associations between patient characteristics and outcome (log-rank test). Cox proportional hazards models were used to determine the multivariable correlations between patient characteristics and outcome. RESULTS: The median follow-up for the docetaxel-based trial (n = 26 patients) was 45 months. Early outcomes for this study are promising. The median disease-free survival (DFS) was 44 months, and the 3-year DFS and overall survival (OS) rates were 58% and 87%, respectively. In the 3 doxorubicin-based studies, the median follow-up was 121.5 months for all living patients, and the estimated 20-year DFS and OS rates were both 26%. On multivariable analysis of patients from all 4 studies, the only significant prognostic factor for DFS and OS (P = 0.0006) was the number of involved axillary lymph nodes at initial diagnosis. CONCLUSIONS: A proportion of patients with isolated BC recurrences achieved prolonged DFS with combined-modality treatment. Patients who receive anthracycline-based chemotherapy at primary diagnosis may benefit from local treatment followed by docetaxel-based chemotherapy for isolated recurrences. The only significant independent prognostic factor was the number of involved axillary lymph nodes at initial diagnosis. PMID- 16047353 TI - Papillary thyroid carcinoma of childhood and adolescence: a 30-year experience at the Istituto Nazionale Tumori in Milan. AB - BACKGROUND: Survival rates are reportedly excellent for papillary thyroid carcinomas (PTCs) in childhood/adolescence, despite their strong tendency to spread. The aim of this study was to verify this assumption in a single institution series spanning a 30-year period with a very long follow-up. PROCEDURE: From 1968 to 2001, 74 cases of thyroid carcinoma were collected. The papillary histological type was confirmed in 42 cases with available slides; we recorded the sex, age at diagnosis, age of menarche, tumor side and size, TNM/pTNM classification, multicentricity, vascular invasion, type of surgery, post-operative complications, post-surgical therapies and outcome up to May 31, 2004. RESULTS: The female/male ratio was 2.2; pT4, pN1 and M1 cases were 52%, 95%, and 12% (four in lungs and one in bone), respectively. Total thyroidectomy was performed in 33 patients, hemithyroidectomy in 8, and a biopsy in 1 inoperable case. Nine patients (21%) relapsed, six in the cervical lymph nodes and three in the lungs. After a median follow-up of 189 months, all patients were alive, two of them with evidence of disease. Overall and progression-free survival curves were independent of sex, age, TNM/pTNM classification, or type of surgery. Overall survival was also independent of recurrence. CONCLUSIONS: Unlike its adult counterpart, PTC of childhood and adolescence is a cancer with a high frequency of spread, but an excellent outcome irrespective of sex, age at diagnosis, TNM/pTNM classification, type of surgery, recurrence. Since pediatric PTCs proved highly responsive to hormone manipulation, it is worth considering a different therapeutic approach from adult cases. PMID- 16047354 TI - Risk factors for diabetes insipidus in langerhans cell histiocytosis. AB - BACKGROUND: Diabetes insipidus (DI) is the most frequent central nervous system (CNS)-related permanent consequence in Langerhans cell histiocytosis (LCH), which mostly requires life-long hormone replacement therapy. In an attempt to define the population at risk for DI, 1,741 patients with LCH registered on the trials DALHX 83 and DALHX 90, LCH I and LCH II were studied. RESULTS: Overall 212 of 1,741 patients (12%) was reported to have DI. In 102 of 1,741 patients (6%) DI was present at diagnosis of LCH. One thousand one hundred eighty three of 1,539 patients without DI at diagnosis had follow up information. One hundred ten of these (9%) later developed DI. The risk of developing DI was 20% at 15 years after diagnosis. Multisystem disease patients at diagnosis carried a 4.6-fold risk for DI compared to single system patients. Craniofacial lesions, in particular in the "ear," "eye," and oral region were associated with a significantly increased risk for DI (relative hazard rate, RHR 1.7), independent of the extent of disease. No influence of the duration of therapy could be determined, but the duration of initial disease activity (RHR 1.5) and the occurrence of reactivations (RHR 3.5) significantly increased the risk for DI. CONCLUSIONS: Patients with multisystem disease and craniofacial involvement at diagnosis, in particular of the "ear," "eye," and the oral region carry a significantly increased risk to develop DI during their course. This risk is augmented when the disease remains active for a longer period or reactivates. PMID- 16047355 TI - Targeted therapy with a cytotoxic somatostatin analog, AN-238, inhibits growth of human experimental endometrial carcinomas expressing multidrug resistance protein MDR-1. AB - BACKGROUND: Chemoresistance mediated by membrane transporters such as multidrug resistance (MDR-1) glycoprotein remains a challenge in the chemotherapy treatment of advanced or recurrent endometrial carcinoma. Targeted chemotherapy might overcome this resistance. The cytotoxic somatostatin (SST) analog, AN-238, consists of a superactive derivative of doxorubicin (DOX), 2-pyrrolino-DOX (AN 201), linked to the SST analog carrier, RC-121. This conjugate binds strongly to SST receptor subtypes (sst) 2a (sst2(a)) and 5 (sst(5)) and can be targeted to tumors that express these receptors. METHODS: The presence of sst2(a) and sst(5) was determined in 3 human endometrial carcinoma cell lines (HEC-1A, RL-95-2, and AN3CA). Nude mice bearing xenografts of these cancers were treated with AN-238 and its radical, AN-201. The antitumor effects and toxicity were compared. The authors studied the effects of AN-238 and AN-201 on the expression levels of MDR 1, multidrug resistance related protein (MRP-1), and breast carcinoma resistance protein (BCRP) by real-time polymerase chain reaction. RESULTS: The authors demonstrated the presence of mRNA and receptor protein for sst(2a) and sst(5) on HEC-1A, RL-95-2, and AN3CA tumors. AN-238 significantly (P < 0.05) inhibited the growth of these tumors, whereas AN-201 had no effect. Blockade of SST receptors nullified the effects of AN-238. In all 3 endometrial carcinoma lines, AN-238 caused a weaker induction of MDR-1 than AN-201. No major induction of MRP-1 and BCRP occurred after treatment with AN-238 or AN-201. CONCLUSIONS: Targeted chemotherapy with the cytotoxic SST analog, AN-238, inhibited powerfully the growth of endometrial carcinoma, which express SST receptors, regardless of their expression level of MDR-1. PMID- 16047356 TI - Few but severe viral infections in children with cancer: a prospective RT-PCR and PCR-based 12-month study. AB - BACKGROUND: Treatment of low-risk febrile episodes with oral administered antibiotics at home is a new approach in pediatric oncology and protective isolation is loosened in more centers. The impact of viral respiratory infections in febrile diseases in this population is still unclear in terms of occurrence and morbidity. PROCEDURE: A prospective follow-up study of all febrile episodes during 12 months in a pediatric oncology department with a high level of protective isolation was set-up with expanded molecular viral examinations. Reverse transcriptase polymerase chain reaction (RT-PCR) and PCR diagnostics of ten viruses, two atypical bacteria, and one fungus were performed and clinical information on all infections was registered. RESULTS: A total of 250 febrile episodes in 66 patients were registered. In all, 198 respiratory secretions, predominantly oral washes, and 165 anal swabs were analyzed. Twenty-two infections were diagnosed: 7 rhinovirus infections, 4 respiratory syncytial virus (RSV) infections, 2 herpes simplex virus (HSV) infections, 2 varicella-zoster virus (VZV) infections, 1 influenza B virus infection, 1 parainfluenza virus type 3 infection (PIV3), 1 human metapneumovirus (HMPV) infection, 1 enterovirus infection, 0 adenovirus infections, 0 influenza A virus infections, and 3 non viral pneumonias: 1 M. Pneumonia, 1 C. Pneumonia, and 1 P. Carinii. The detected pathogens correlated well to the clinical disease. Patients with viral infections were as affected as patients with bacteria in the blood. One of 19 viral infections was lethal, a RSV pneumonia. C-reactive protein concentrations were not able to distinguish viral infections from bacteremias. CONCLUSIONS: The applied sampling method was acceptable and molecular diagnosis of viruses, atypical bacteria and P. Carinii increased the microbiological verification of infections by 35%. Viral infections were few in our protected population but caused severe infectious complications in these patients. PMID- 16047357 TI - Successful treatment of the HIV-associated Burkitt lymphoma in a three-year-old child. AB - We report a case of successful treatment of advanced Burkitt lymphoma in a 3-year old girl with congenitally acquired HIV infection. She was treated with intensive chemotherapy combined with highly active anti-retroviral therapy (HAART). She had not received any anti-retroviral therapy and had stage C3 HIV infection on admission. She was treated with modified BFM 90 and remission was achieved. Simultaneously with chemotherapy, HAART with three drugs was carried out, resulting in improvements in HIV parameters. Our report suggests chemotherapy combined with HAART is feasible in children and may favorably change the outcomes in pediatric patients with AIDS-NHL. PMID- 16047358 TI - Intraoperative bile cytology of the dysplasia-carcinoma in situ sequence of gallbladder carcinoma. AB - BACKGROUND: The preoperative diagnosis of noninvasive and early carcinoma of the gallbladder is considered to be both difficult and exceptional. In the current study, the authors conducted a cytologic analysis of bile in the intraoperative diagnosis of clinically unsuspected gallbladder carcinoma. METHODS: Bile samples collected from 40 patients at the time of elective cholecystectomy were centrifuged. Smears prepared from the sediments were stained with May-Grunwald Giemsa and Papanicolaou stains. Lesions were categorized as hyperplasia with or without metaplasia, dysplasia, carcinoma in situ, and invasive carcinoma. Cytologic diagnoses were compared with histopathology. RESULTS: Two cases of carcinoma in situ and one case of invasive carcinoma diagnosed on bile cytology were confirmed by histopathology. No false-positive diagnoses were made based on cytologic examination of aspirated bile. Cytohistologic concordance in cases of hyperplasia, dysplasia, and chronic cholecystitis varied from 56-60%. CONCLUSIONS: Intraoperative bile cytology was found to be a reliable method for the detection of in situ and early invasive carcinoma of the gallbladder. PMID- 16047359 TI - Temozolomide and oral VP-16 for children and young adults with recurrent or treatment-induced malignant gliomas. AB - BACKGROUND: Children and young adults with recurrent or treatment-induced malignant gliomas have limited responses to temozolomide or oral VP-16 when either is administered as a single agent. We postulated that a combination of these two drugs for patients with recurrent or treatment-induced malignant gliomas might result in better and more prolonged responses. A retrospective analysis was performed on patients treated with the combination of temozolomide and VP-16. PROCEDURE: Eleven patients with recurrent or treatment-induced malignant gliomas were treated with varying combinations of temozolomide (150-210 mg/m2/d for 5 days) and oral VP-16 (50 mg/m2/d for 4-12 days). Responses were assessed by MRI scan, and data on clinical course and toxicity were retrospectively obtained from the medical record. RESULTS: The median age of the 11 patients was 17 years (range 5-23 years). Diagnoses included recurrent brain stem glioma (2), recurrent anaplastic astrocytoma (2), and glioblastoma (7) (3 treatment-induced, 2 malignant transformations of lower grade tumors, 1 recurrence, and 1 second tumor arising 10 months after diagnosis of medulloblastoma). All 11 patients had received radiotherapy (including 4 who received craniospinal radiation), and 7 had prior chemotherapy. Nine patients were treated at first recurrence, two at second recurrence. One patient had a complete response (CR), six had partial responses (PR), and four had progressive disease (PD). The median progression-free survival for the seven responding patients was 6 months (range 4-15+ months). There was one grade 4 neutropenia, but no other grade 3 or 4 toxicities. CONCLUSIONS: These data suggest there is activity of temozolomide in combination with oral VP-16 for children and young adults with recurrent malignant gliomas. PMID- 16047360 TI - M.TaqI facilitates the base flipping via an unusual DNA backbone conformation. AB - MD simulations have been carried out to understand the dynamical behavior of the DNA substrate of the Thermus aquaticus DNA methyltransferase (M.TaqI) in the methylation process at N6 of adenine. As starting structures, an x-ray structure of M.TaqI in complex with DNA and cofactor analogue (PDB code: 1G 38) and free decamer d(GTTCGATGTC)(2) were taken. The x-ray structure shows two consecutive BII substates that are not observed in the free decamer. These consecutive BII substates are also observed during our simulation. Additionally, their facing backbones adopt the same conformations. These double facing BII substates are stable during the last 9 ns of the trajectories and result in a stretched DNA structure. On the other hand, protein-DNA contacts on 5' and 3' phosphodiester groups of the partner thymine of flipped adenine have changed. The sugar and phosphate parts of thymine have moved further into the empty space left by the flipping base without the influence of protein. Furthermore, readily high populated BII substates at the GpA step of palindromic tetrad TCGA rather than CpG step are observed in the free decamer. On the contrary, the BI substate at the GpA step is observed on the flipped adenine strand. A restrained MD simulation, reproducing the BI/BII pattern in the complex, demonstrated the influence of the unusual backbone conformation on the dynamical behavior of the target base. This finding along with the increased nearby interstrand phosphate distance is supportive to the N6-methylation mechanism. PMID- 16047361 TI - Temozolomide in resistant or relapsed pediatric solid tumors. AB - PURPOSE: We report the off-label study aimed at investigating the use of temozolomide (TMZ) as single agent in relapsed or resistant pediatric solid tumors. The drug was administered at the dose of 215 mg/m2/day x 5 days or 180 mg/m2/day x 5 days in patients with prior craniospinal irradiation (CSI) or autologous bone marrow transplantation (ABMT). PATIENTS AND METHODS: Fifty two patients, median age 127.6 months, with resistant or relapsed solid tumors were enrolled. Tumor types were: neuroblastoma (NB; n = 17), medulloblastoma (MB; 8), brain stem glioma (BSG; 8), extraosseous Ewing's sarcoma/peripheral neuroectodermal tumor (EOES; 4), Ewing's sarcoma (ES; 4), anaplastic astrocytoma (AA; 3), rhabdomyosarcoma (RMS; 2), ependymoma (EP; 2), cerebral primitive neuroectodermal tumor (cPNET; 2), hepatocarcinoma (HC; 1), and osteosarcoma (OS; 1). All patients were pre-treated. Two outpatient courses were administered, with a median of 4.8 courses/pt. RESULTS: Objective response-rate (CR + PR + MR) in our series was 13.4% (1.9% CR, 3.8% PR, and 7.7% MR), SD occurred in 38.4% of patients and 48% had PD. The median survival was 7.8 months (range 1-37) and median time to progression was 3.4 months (range 1-20); these data were significantly correlated with histology and previous nitrosureas administration in multivariate analysis. Haematological toxicity grade 3-4 (mainly thrombocytopenia) was observed in 21.4% of administered courses, nausea was reported in 3.1% and respiratory distress in 0.7%. CONCLUSION: Oral TMZ was well tolerated in children with resistant or relapsed solid tumors and showed activity in NB and CNS tumours refractory to standard chemotherapy. PMID- 16047362 TI - Alternative medicine remedies might stimulate viability of leukemic cells. AB - OBJECTIVE: Ex vivo activity of three products of alternative therapy against leukemic and normal cells was analyzed. MATERIAL AND METHODS: Extracts of Viscum album, Uncaria tomentosa, and Croton lechleri were used for the study. Leukemic cells of 53 children with acute leukemias and four cell lines, Jurkat, CCRF-CEM, HL-60, and K-562 were tested for sensitivity to alternative medicine remedies by the MTT assay, cell-cycle analysis and annexin-V binding assay. RESULTS: Leukemic cells showed high resistance to tested three compounds of alternative medicine in all performed assays. Additionally, tested remedies stimulated survival of leukemic cells in 45%, 96%, and 83% cases, respectively; while no effect was observed in normal lymphocytes. In combination studies, Viscum album extract did not increase prednisolone and cytarabine cytotoxicity in leukemic cells. CONCLUSION: We conclude that some alternative medicine remedies might stimulate the viability of childhood leukemic cells. PMID- 16047363 TI - Intussusception in children with ALL receiving chemotherapy for acute lymphoblastic leukaemia. AB - Acute abdominal complications of chemotherapy are common but the differential diagnosis wide. We describe two cases of intussuception of the bowel in children receiving chemotherapy for acute lymphoblastic leukaemia (ALL) and discuss how a high clinical suspicion is critical for the correct diagnosis to be made rapidly. PMID- 16047364 TI - The role of radiotherapy in the treatment of childhood intracranial germinoma: long-term survival and late effects. AB - BACKGROUND: The aim of the present report was to evaluate the role of radiotherapy in the treatment of childhood intracranial germinoma in view of long term survival and functional outcome. PROCEDURE: Nine children with histologically verified intracranial germinomas treated in Slovenia between 1983 and 1995 were reviewed. The four boys and five girls were 8.8-16.9 years old (median, 11.3 years). Five tumors were suprasellar, three were in the pineal region, and one patient had bifocal disease. Two patients had disseminated tumor. All patients received radiotherapy: six to the tumor bed, one to the whole brain, and two to the whole central nervous axis (CNA). The doses to the tumor bed ranged from 30 to 46 Gy (median, 44 Gy) and to the CNA were 24 and 34.5 Gy. Five patients received neoadjuvant cyclophosphamide and three patients, all with beta human chorionic gonadotropin secreting tumors, received neoadjuvant cisplatin based chemotherapy. RESULTS: Six patients are alive 12.8-21.8 years (median, 19 years) from diagnosis. The causes of death in three patients were disseminated disease, toxicity of salvage chemotherapy, and secondary etoposide-induced leukemia. All patients with suprasellar tumors presented with overt endocrinopathy. Results of psychological evaluation were subnormal in one out of five patients tested. Estimate of mental deterioration due to therapy ranged from 0% to 30% (median, 15%). Emotional disorder was registered in four patients and psycho-organic syndrome in three. CONCLUSIONS: Our results on long-term survival and functional outcome confirm the efficacy and relative safety of limited-field and reduced-dose radiotherapy for childhood intracranial germinoma when supplemented with chemotherapy. PMID- 16047365 TI - Quantitative assessment of ventricular function in sickle cell disease: effect of long-term erythrocytapheresis. AB - BACKGROUND: Previous studies on cardiac function in patients with sickle cell disease (SCD) demonstrated abnormalities of systolic and diastolic function including elevated left ventricular myocardial performance index (LVMPI) on chronic transfusion protocols. LVMPI has been validated as a useful and easy non invasive measure of overall cardiac function. Up to now, there are no reported studies on cardiac function in patients with SCD maintained on long-term erythrocytapheresis (LTE). PROCEDURES: We recorded LVMPI in 22 patients with SCD aged 3-20 years and we compared the results between non-transfused patients (NT SCD) and patients on LTE (T-SCD). RESULTS: Males with SCD had higher mean LVMPI than females (P = 0.04). There were significant differences among T-SCD, severe NT-SCD, and mild NT-SCD patients with respect to hemoglobin (Hb) levels (P = .003) and TR velocity (P = .03). T-SCD patients showed elevated LVMPI compared to NT-SCD patients with severe and mild disease (P = 0.002). Pair-wise comparisons demonstrated that T-SCD patients had LVMPI that was significantly higher than NT SCD (mild) patients (P = 0.01). CONCLUSIONS: Our study demonstrates that patients SCD on LTE have cardiac dysfunction based on elevated LVMPI. This may be a reflection of the global severity of disease. Our findings merit further investigation with serial monitoring of LVMPI on a larger number of patients with SCD. PMID- 16047366 TI - Paraneoplastic syndrome and intrathoracic Castleman disease. AB - We report two cases of intrathoracic Castleman disease presenting with paraneoplastic syndrome. Patient 1 was a 10-year-old girl with short stature. She was found to have delayed bone age, slow growth velocity, and iron-deficiency anemia, which was refractory to treatment. Thrombocytosis and hypergammaglobulinemia were later detected. Chest X-ray revealed a hilar mass. Patient 2 was a 14-year-old boy who had severe cough, progressive mucocutaneous erosion, and dermatitis. Chest X-ray showed a mediastinal mass. Sections of skin biopsy showed findings consistent with pemphigus disease. In each case, the histological diagnosis of Castleman disease was made. PMID- 16047367 TI - Intracranial germinomas: can we improve upon our success? PMID- 16047368 TI - Sentinel node biopsy in squamous cell cancer of the oral cavity and oral pharynx: a diagnostic meta-analysis. AB - BACKGROUND: The sentinel node biopsy concept has been gaining support in the head and neck cancer literature during only the last few years, and several pilot studies have been published. This procedure aims to avoid unnecessary treatment to the clinically negative neck by identifying the patients with occult neck disease. METHODS: We performed a systematic review and a diagnostic meta-analysis of all published literature regarding sentinel node biopsies in head and neck cancer until December 2003 using established guidelines. Using the pooled sensitivity rates obtained from the meta-analysis and treatment outcomes from published literature, we created a decision analysis model to identify the treatment arm with better payoffs. RESULTS: A total of 301 patients with oral cavity primary tumors and 46 patients with oropharyngeal primary tumors from 19 articles were included for the meta-analysis. The pooled sensitivity result using the random effects model was 0.926 (95% confidence interval, 0.852-0.964). The cumulative payoff for the sentinel node biopsy arm was lower than that for the elective node dissection arm by about 1%. The payoffs were assigned for the recurrence and mortality rates only and did not take into account the morbidity caused by the procedures. CONCLUSIONS: The sentinel node biopsy procedure has shown high sensitivity rates in pilot studies for oral and oropharyngeal squamous cell cancer across the globe and is reliable and reproducible. This study provides a firm evidence base for forthcoming trials on the role of sentinel node biopsy in head and neck cancer. PMID- 16047369 TI - Uncommon presentation of craniopharyngioma with anemia in an adolescent. PMID- 16047370 TI - Toxicity due to 6-thioquanine. PMID- 16047371 TI - Enhanced proliferation of progenitor cells in the subventricular zone and limited neuronal production in the striatum and neocortex of adult macaque monkeys after global cerebral ischemia. AB - Cerebral ischemia in adult rodent models increases the proliferation of endogenous neural progenitor cells residing in the subventricular zone along the anterior horn of the lateral ventricle (SVZ a) and induces neurogenesis in the postischemic striatum and cortex. Whether the adult primate brain preserves a similar ability in response to an ischemic insult is uncertain. We used the DNA synthesis indicator bromodeoxyuridine (BrdU) to label newly generated cells in adult macaque monkeys and show here that the proliferation of cells with a progenitor phenotype (double positive for BrdU and the markers Musashi 1, Nestin, and beta III-tubulin) in SVZ a increased during the second week after a 20-min transient global brain ischemia. Subsequent progenitor migration seemed restricted to the rostral migratory stream toward the olfactory bulb and ischemia increased the proportion of adult-generated cells retaining their location in SVZ a with a progenitor phenotype. Despite the lack of evidence for progenitor cell migration toward the postischemic striatum or prefrontal neocortex, a small but sustained proportion of BrdU-labeled cells expressed features of postmitotic neurons (positive for the protein Neu N and the transcription factors Tbr 1 and Islet 1) in these two regions for at least 79 days after ischemia. Taken together, our data suggest an enhanced neurogenic response in the adult primate telencephalon after a cerebral ischemic insult. PMID- 16047372 TI - Treatment of disseminated aspergillosis with voriconazole/liposomal amphotericin B in a child with leukemia. PMID- 16047373 TI - Pulmonary embolism due to invasive aspergillosis in a child with acute myelogenous leukemia. PMID- 16047374 TI - Shwachman-Diamond syndrome. AB - Shwachman-Diamond syndrome (SDS) is an inherited marrow failure disorder with varying cytopenia, pancreatic dysfunction, and metaphyseal dysostosis. SDS is also characterized by a risk of myelodysplasia and leukemia in up to one third of the patients. Over the last 5 years, major advances have been made in understanding the bone marrow phenotype. The gene associated with the disease, SBDS, has recently been identified. Herein we provide an update on the clinical features, the hematopoietic defects, and the genetics of the disease as they are currently understood. We also review the diagnostic and therapeutic approaches to the hematological complications in the syndrome. PMID- 16047375 TI - Contrast-enhanced power Doppler sonography of malignant ovarian tumors using harmonic flash-echo imaging: preliminary experience. AB - To investigate the potential usefulness of contrast-enhanced intermittent harmonic sonography in the diagnosis of ovarian cancer, we evaluated 4 patients with complex adnexal masses suspected of malignancy using intermittent harmonic sonography after injection of a contrast agent. Tumor and/or mural nodule tissue enhancement was detected in all cases of ovarian malignancy. Contrast-enhanced, intermittent harmonic sonography provides a satisfactory visualization of blood flow in the solid portion of the tumor tissue and may support a diagnosis of ovarian malignancy. Depiction of blood vessels using low MI techniques may be possible with other vascular ultrasonographic contrast agents. PMID- 16047376 TI - Hemimegalencephaly: cranial sonographic findings in neonates. AB - Hemimegalencephaly is a rare neuronal migration disorder characterized by overgrowth of part or all of 1 cerebral hemisphere. Lissencephaly, pachygyria, polymicrogyria, heterotopia, and glial abnormalities are usually noted pathologically in the enlarged hemisphere. We report the sonographic changes associated with hemimegalencephaly in 2 neonates presenting with seizures and correlate these changes with the magnetic resonance imaging findings. PMID- 16047377 TI - Splenic vein aneurysm with calcification of splenic and portal veins. AB - Splenic vein aneurysm is a rare disease, and calcifications in the portal venous system are also rare. Here we present a case of splenic vein aneurysm associated with calcification of the splenic and portal veins, a condition that to our knowledge has not been reported before. PMID- 16047378 TI - Multilocular adenomatoid tumor of the ovary: ultrasonographic findings. AB - We report the sonographic findings of a rare benign ovarian tumor in a 69-year old woman. Transvaginal ultrasonography showed a cystic multilocular lesion with a vascularized central solid portion of the left ovary. Surgery revealed an adenomatoid tumor. Adenomatoid tumors are benign lesions of mesothelial origin, usually solid in nature and rarely located in the ovaries. (c) 2005 Wiley Periodicals, Inc. J Clin Ultrasound 33:233-236, 2005. PMID- 16047379 TI - Combined segmental and focal adenomyomatosis involving the body of the gallbladder. AB - Adenomyomatosis (adenomyomatous hyperplasia) of the gallbladder is a benign process of unknown origin that has been classified into three morphologic types: diffuse, segmental, and focal (fundal). Cases of combined (segmental and focal) forms involving the gallbladder body are very rare. We present the sonographic findings of a case having adenomyomatosis of the gallbladder body with focal as well as segmental forms of the disease. PMID- 16047380 TI - Evaluating physical function in an adolescent bone tumor population. AB - BACKGROUND: Survival rates for patients with bone tumors have increased dramatically over the past few decades. Unfortunately, many patients face functional limitations resulting from disease management, but there is little evidence regarding physical functioning in adolescents with bone tumors. This lack is largely due to the absence of a standardized measure for evaluating adolescent sarcoma physical functioning. This review discusses considerations in searching for a measure of adolescent physical functioning, evaluates measures previously used in assessing functional outcome in young sarcoma patients, reviews self-report measures of pediatric physical functioning, and makes recommendations for future research. PROCEDURE: Electronic databases were searched for studies published between 1980 and 2004. Studies examining physical functioning in adolescent sarcoma patients and those describing measures of pediatric physical functioning were included. RESULTS: The study includes measures previously used in the adolescent bone tumor literature (Musculoskeletal Tumor Society, Mankin, Groningen Activity Restriction Scale, Toronto Extremity Salvage Score [TESS]), and three measures of pediatric physical function (Functional Disability Inventory, Pediatric Outcomes Data Collection Instrument [PODCI], Activities Scale for Kids [ASK]). The TESS, PODCI, and ASK each met at least four of the outlined applicability and feasibility components. CONCLUSION: The search for a measure of physical functioning for adolescent sarcoma patients requires an exploration of many developmental and measurement issues. Based on the sensibility review presented, the TESS, PODCI, and ASK are recommended for further evaluation of content and of the measurement properties of reliability, validity, and responsiveness, prior to their use in evaluating adolescent sarcoma patients. PMID- 16047381 TI - Role of IGFBPs in the morphogenesis of lingual papillae. AB - The expression of insulin-like growth factor binding proteins (IGFBPs) during the morphogenesis of lingual papillae of mice was examined by in situ hybridization. Among seven mouse IGFBPs, IGFBP-1, -6, and -7 mRNAs were not expressed in the tongue tissue. At E12, though no papillae have formed yet, IGFBP-2, -4, and -5 were expressed in the entire tongue epithelium. At E14, fungiform papillae appeared in the anterior region and circumvallate papillae were distinguished in the posterior region. Strong expression of IGFBP-5 was observed in the apical region of both fungiform and circumvallate papillae. At this stage, the epithelial elevation of filiform papillae was not clear; but IGFBP-5 was expressed in the apex. At E15, foliate papillae were distinguished and IGFBP-5 was expressed in the dorsal epithelium of ridges. In filiform papillae, IGFBP-3 was expressed in the core of the connective tissue. At E17, the expression of IGFBP-5 disappeared from the apical region of fungiform, filiform, foliate, and circumvallate papillae, whereas that of IGFBP-2 remained. This finding suggests that IGFBP-5 and -2 function to cause evagination of the epithelium into a raised structure. In the epithelium of trenches of foliate and circumvallate papillae, strong expression of IGFBP-4 was observed at E15 and E17. As previously suggested from a study on postnatal mice (Suzuki et al. J Comp Neurol 2005;482:74-84), IGFBP-4 acts in the epithelial invagination to form the trenches, grooves, or furrows of lingual papillae during development. PMID- 16047382 TI - Osteoblasts engulf apoptotic bodies during alveolar bone formation in the rat maxilla. AB - During bone formation, as in other tissues and organs, intense cellular proliferation and differentiation are usually observed. It has been described that programmed cell death, i.e., apoptosis, takes place in the control of the cellular population by removing of the excessive and damaged cells. Although it is generally accepted that apoptotic bodies are engulfed by professional phagocytes, the neighboring cells can also take part in the removal of apoptotic bodies. In the present study, regions of initial alveolar bone formation of rat molars were examined with the aim to verify whether osteoblasts are capable of engulfing apoptotic bodies, such as professional phagocytes. Rats aged 11-19 days were sacrificed and the maxillary fragments containing the first molar were removed and immersed in the fixative solution. The specimens fixed in glutaraldehyde-formaldehyde were processed for light microscopy and transmission electron microscopy. For the detection of apoptosis, the specimens were fixed in formaldehyde, embedded in paraffin, and submitted to the TUNEL method. The results revealed round/ovoid structures containing dense bodies on the bone surface in close contact to osteoblasts and in conspicuous osteoblast vacuoles. These round/ovoid structures showed also positivity to the TUNEL method, indicating that bone cells on the bone surface are undergoing apoptosis. Ultrathin sections showed images of apoptotic bodies being engulfed by osteoblasts. Occasionally, the osteoblasts exhibited large vacuoles containing blocks of condensed chromatin and remnants of organelles. Thus, these images suggest that osteoblasts are able to engulf and degrade apoptotic bodies. PMID- 16047383 TI - LEK1 protein expression in normal and dysregulated cardiomyocyte mitosis. AB - A defining characteristic of embryonic cells is their ability to divide rapidly, even in tissues such as cardiac muscle, which cannot divide once fully differentiated. This suggests that regulators of cell division differ in embryonic and differentiated cells. LEK1 is a member of an emerging family of proteins with diverse functions but shared structural domains, including numerous leucine zippers, a nuclear localization site, and a functional Rb-binding domain. LEK1 is expressed ubiquitously in the developing mouse embryo from the earliest stages of differentiation through birth. It is absent in adult tissues, even those that maintain active cell division. We hypothesize that LEK1 is a regulator of mitosis restricted to the developing embryo and early neonate. Here, using BrdU incorporation, we show that LEK1 protein downregulation in cardiac myocytes correlates directly with cessation of DNA synthesis between neonatal days 6 and 10. In contrast, in an immortalized cardiac cell line (HL1 cells), both BrdU incorporation and LEK1 protein expression persist, and actively dividing cells express LEK1. However, BrdU incorporation can be decreased in these cells by treatment with a morpholino targeting LEK1 mRNA. These data suggest a role for LEK1 in regulating the normal embryonic cardiomyocyte cell cycle and in promoting continued mitosis in transformed, abnormally dividing cardiomyocytes. PMID- 16047384 TI - Proteomic comparison of two fractions derived from the transsynaptic scaffold. AB - A fraction derived from presynaptic specializations (presynaptic particle fraction; PPF) can be separated from postsynaptic densities (PSD) by adjusting the pH of Triton X-100 (TX-100) extraction of isolated transsynaptic scaffolds. Solubilization of the PPF corresponds to disruption of the presynaptic specialization. We show that the PPF is insoluble to repeated TX-100 extraction at pH 6.0 but becomes soluble in detergent at pH 8.0. By immunolocalization, we find that the major proteins of the PPF, clathrin and dynamin, are concentrated in the presynaptic compartment. By using multidimensional protein identification technology, we compared the protein compositions of the PPF and the PSD fraction. We identified a total of 341 proteins, 50 of which were uniquely found in the PPF, 231 in the PSD fraction, and 60 in both fractions. Comparison of the two fractions revealed a relatively low proportion of actin and associated proteins and a high proportion of vesicle or intracellular compartment proteins in the PPF. We conclude that the PPF consists of presynaptic proteins not connected to the actin-based synaptic framework; its insolubility in pH 6 and solubility in pH 8 buffered detergent suggests that clathrin might be an anchorage scaffold for many proteins in the PPF. PMID- 16047385 TI - CCL2 and CXCL1 trigger calcitonin gene-related peptide release by exciting primary nociceptive neurons. AB - Chemokines are important mediators in immune responses and inflammatory processes. Calcitonin gene-related peptide (CGRP) is produced in dorsal root ganglion (DRG) neurons. In this study, CGRP radioimmunoassay was used to investigate whether the chemokines CCL2 and CXCL1 could trigger CGRP release from cultured DRG neurons of neonatal rats and, if so, which cellular signaling pathway was involved. The results showed that CCL2 and CXCL1 ( approximately 5 100 ng/ml) evoked CGRP release and intracellular calcium elevation in a pertussis toxin (PTX)-sensitive manner. The CGRP release by CCL2 and CXCL1 was significantly inhibited by EGTA, omega-conotoxin GVIA (an N-type calcium channel blocker), thapsigargin, and ryanodine. Pretreatment of DRG neurons for 30 min with the inhibitors of phospholipase C (PLC) and protein kinase C (PKC) but not mitogen-activated protein kinases (MAPKs) significantly reduced CCL2- or CXCL1 induced CGRP release and intracellular calcium elevation. Intraplantar injection of CCL2 or CXCL1 produced hyperalgesia to thermal and mechanical stimulation in rats. These data suggest that CCL2 and CXCL1 can stimulate CGRP release and intracellular calcium elevation in DRG neurons. PLC-, PKC-, and calcium-induced calcium release from ryanodine-sensitive calcium stores signaling pathways are involved in CCL2- and CXCL1-induced CGRP release from primary nociceptive neurons, in which chemokines produce painful effects via direct actions on chemokine receptors expressed by nociceptive neurons. PMID- 16047386 TI - Effects of exercise on gallbladder volume and motility in obese women. AB - PURPOSE: Comparatively few prospective studies have investigated the relationship between physical activity and gallbladder motility, and the results are controversial. Exercise may affect gallbladder motility via neural or hormonal mechanisms. The purpose of this study was to evaluate the possible effects of aerobic exercise on gallbladder motility in a group of obese women without gallstones. PATIENTS AND METHODS: Twenty-three obese women (age 41.2+/-10.3 years, body mass index 40.7+/-6.7 kg/m(2)) were included in the study. Following an overnight fast, fasting and postprandial (15, 30, 45, 60, 75, 90, 120, and 150 minute) volumes and ejection fractions were evaluated with real-time ultrasonography before exercise. For all subjects, the exercise regimen consisted of daily 45-minute walking sessions at 60-80% of maximum heart rate for 4 weeks except weekends. Gallbladder volume and ejection fraction were again evaluated after exercise. RESULTS: Fasting and postprandial (15, 30, 45, 60, 75, 90, 120, and 150 minute) volumes were 38.6+/- 10.9, 32.8+/- 8.8, 27.6/- 8.1, 22.7+/- 8.5, 21.4+/- 7.2, 20.8+/- 7.0, 22.8+/- 7.3, 29.6 +/- 7.0, and 36.8+/- 6.2 cm(3) before the exercise period, respectively, and 40.8+/- 18.9, 29.9+/-11.2, 25.3+/- 9.2, 22.4+/-8.5, 19.6+/-7.8, 17.7+/- 6.8, 17.8+/- 7.3, 23.1+/-10.8, and 29.0+/-14.4 cm(3) after the exercise period, respectively. Postprandial (15, 30, 45, 60, 75, 90, 120, and 150 minute) ejection fractions were 13.5+/-15.9, 27.4+/-15.4, 39.5+/ 20.0, 43.2+/-16.7, 44.3+/-17.3, 37.5 +/- 23.5, 23.5 +/-25.1, and 5.5+/- 21.6% before the exercise period, respectively, and 22.6+/- 20.1, 34.6+/-14.5, 42.0+/ 13.6, 49.2+/-12.6, 53.1+/-14.1, 52.6+/-16.1, 43.6+/-17.0, and 29.2+/- 26.5% after exercise, respectively. After the exercise period, the 75, 90, 120, and 150 minute volumes were lower (p< 0.05, p< 0.05, p< 0.05, p< 0.01) and the 90, 120, and 150 minute ejection fractions were higher than before exercise (p< 0.05, p < 0.05, p< 0.01). CONCLUSIONS: Our study showed that exercise decreased late-phase postprandial gallbladder volume and increased late-phase postprandial gallbladder motility in these obese women. PMID- 16047387 TI - Using fetal acoustic stimulation to shorten the biophysical profile. AB - PURPOSE: To determine whether fetal acoustic stimulation can decrease the time required to achieve a reassuring biophysical profile. METHODS: Patients scheduled for a biophysical profile were prospectively assigned to study and control groups. The study group received 3 seconds of acoustic stimulation if fetal breathing, tone, or movement were not present during the first 5 minutes of the study. The biophysical profile was completed in the standard fashion, for both groups. RESULTS: A total of 870 patients were enrolled (458 control, 412 with stimulation). The fetal acoustic stimulation group had decreased testing time (3 minutes) and fewer non-reassuring tests (5%). The fetal acoustic stimulation group had fewer studies without breathing, potentially reducing the need for further testing or intervention. CONCLUSIONS: Fetal acoustic stimulation can be used to decrease the biophysical profile testing time and to reduce the number of non-reassuring tests. PMID- 16047388 TI - Dynamic ultrasonography of rotator cuff muscles. AB - PURPOSE: To confirm the neuromuscular integrity of the rotator cuff and to investigate the effect of different contraction patterns on shoulder function, we sonographically analyzed dynamic contraction patterns of this muscle group. METHODS: Fifty supra- and 50 infraspinatus muscle contraction patterns of patients with different shoulder pathologies examined with a 7.5-MHz transducer were recorded and saved as video files. Both data sets were analyzed by 3 independent observers who then assigned a grade to each contraction pattern: normal, slightly disturbed, severely disturbed, or no contraction. Intra- and inter-observer variations were calculated. In 43 patients with isolated supraspinatus tears, the clinical relation to shoulder function as measured with the Constant score was evaluated. RESULTS: In the 100 cases tested, the intraclass correlations for the 3 observers were 0.82, 0.88, and 0.88. The inter observer reliability was 0.74. No pair of first and second readings for either the supraspinatus or infraspinatus muscle differed by more than 1 grade. In patients with a supraspinatus tear, the contraction type of the supraspinatus correlated significantly with the Constant score (p=0.02). CONCLUSIONS: When dynamic ultrasonography is used to assess the contraction patterns of the supra- and infraspinatus muscles, good intra- and inter-observer reliability is attained. Because a better contraction type correlates significantly with better shoulder function, this new diagnostic criterion may improve decision-making in the treatment of shoulder diseases. PMID- 16047389 TI - Volume changes in the gallbladder in association with various radiologic examinations. AB - PURPOSE: Our aim was to evaluate with sonography the volume changes of the gallbladder after intravenous administration of nonionic, iodinated contrast agents with or without ingestion of contrast agent or water. METHODS: A total of 191 patients received intravenous injection of 50 ml of nonionic contrast agent (Group A), intravenous injection of 100 ml of contrast medium (Group B), intravenous injection of 100 ml of contrast medium together with ingestion of 1000 ml of a dilute solution of contrast medium (Group C), or 1000 ml of water taken orally (Group D). The gallbladder volume was measured by ultrasonography in the fasting state in all groups, 15 minutes after intravenous injection of contrast medium in Groups A, B, and C and 30 minutes after water ingestion in Group D. RESULTS: In all groups, the volume of the gallbladder was statistically lower after the procedure than before. There was no significant difference among the groups in terms of this gallbladder volume change. CONCLUSIONS: Intravenous injection of nonionic, iodinated contrast medium or ingestion of water or a dilute solution of contrast medium for various radiological examinations results in contraction of the gallbladder. Contrast-enhanced radiological examinations may interfere with sonographic evaluation of the gallbladder. If necessary, gallbladder ultrasonographic examination should be repeated with the patient in a fasting state. (c) 2005 Wiley Periodicals, Inc. J Clin Ultrasound 33:214-217, 2005. PMID- 16047390 TI - Endosonographic evaluation of perianal fistulas and abscesses: comparison of two instruments and assessment of the role of hydrogen peroxide injection. AB - PURPOSE: The aims of this study were to compare the performance of a mechanical radial endosonoscope and an endorectal electronic probe and to evaluate the value of hydrogen peroxide (H(2)O(2)) injection in the diagnosis of perianal fistulas. PATIENTS AND METHODS: Sixty-one patients underwent clinical and endosonographic studies for suspected perianal fistulas or abscesses. Endosonography was performed using two instruments: a mechanical radial endosonoscope (Olympus GF UM20) and an electronic endorectal probe (Hitachi EUP-R53). The patients were re examined during and following H(2)O(2) injection using both systems. RESULTS: Thirty-seven fistulous tracts were visualized with the electronic endorectal probe versus only 9 with the mechanical radial endosonoscope. Four patients had anal stenosis, precluding the use of the larger electronic probe. Three fistulas were detected in these patients using the mechanical radial endosonoscope. H(2)O(2) injection was not feasible in 26 patients (43%). Visualization of 11 (31%) fistulas improved after administration of H(2)O(2). Six fistulous tracts not detected before H(2)O(2) administration were clearly visualized during injection and for several minutes thereafter. CONCLUSIONS: Work-up of perianal fistula should be preferably performed using the endorectal electronic probe. However, the mechanical radial endoscope is preferred in patients with anal stricture. H(2)O(2) injection should become an integral part of every sonographic study of perianal fistulas. PMID- 16047391 TI - Production of inbred and hybrid transgenic mice carrying large (> 200 kb) foreign DNA fragments by intracytoplasmic sperm injection. AB - We have developed a mouse transgenesis technique that facilitates the insertion of large (approximately 200 kilo base pairs) DNA fragments into host genomes of both inbred and hybrid mice. Six inbred and three hybrid transgenic mice carrying a single bacterial artificial chromosome (BAC) clone with genes located in the Down syndrome critical region of human chromosome 21 were produced using this technology. PMID- 16047392 TI - Mouse zygotes with one diploid pronucleus formed as a result of ICSI can develop normally beyond birth. AB - A mouse spermatozoon was injected into mouse secondary oocytes (ICSI) in the vicinity of the metaphase spindle. In 22% of oocytes injected successfully, the maternal chromatin (the haploid chromatids formed after the second meiotic division) and paternal chromatin (from the sperm nucleus) were surrounded by a common nuclear envelope to form one diploid bi-parental pronucleus. However, the use of spermatozoa in which BrdU had been incorporated into DNA during spermatogenesis revealed, that maternal and paternal chromatin occupied two separate compartments within the one pronucleus. In the living state, the diploid pronucleus could be distinguished from a haploid one by its distinctly larger size and by a greater number of "nucleolus-like bodies"-criteria confirmed karylogically at the 1st cleavage division. Such zygotes with one diploid pronucleus were able to develop in vitro into blastocysts as often as those with two haploid pronuclei [11/29 (38%) vs. 14/35 (40%)]. Seventy nine 2-cell embryos developing in vitro from zygotes with one diploid pronucleus were transplanted to the oviducts of pseudopregnant recipients: two females had six foetuses when killed on the 17th day, and two females gave birth to nine young, eight of which survived and developed into normal fertile animals. PMID- 16047393 TI - The breakdown of the minimum polarizability principle in vibrational motions as an indicator of the most aromatic center. AB - The vibrational motions that disobey the minimum polarizability principle (MPP) in pi-conjugated molecules are distortions of the equilibrium geometry that produce a reduction in the polarizability due to the localization of pi electrons. For aromatic species, this electronic localization is responsible for the subsequent reduction in the aromaticity of the system. In the present work, we diagonalize the Hessian matrix of the polarizability with respect to the vibrational nontotally symmetric normal coordinates, to calculate the nontotally symmetric distortions that produce the maximum breakdown of the MPP in a series of twenty polycyclic aromatic hydrocarbons. It is shown that the nuclear displacements that break the MPP have larger components in those rings that possess the highest local aromaticity. Thus, these vibrational motions can be used as an indicator of local aromaticity. PMID- 16047394 TI - Mono- and dinuclear bioxazoline-palladium complexes for the stereocontrolled synthesis of CO/styrene polyketones. AB - The coordination chemistry of the chiral bioxazoline ligand (4S,4'S)-2,2'-bis(4 isopropyl-4,5-dihydrooxazole) to Pd(II) provides evidence that the ligand bonding can occur either through chelation of one Pd(II) ion leading to a mononuclear species with the expected cis geometry, or by double bridging of two Pd(II) ions giving a dinuclear complex with trans geometry. The species in solution are identified by 1H NMR spectroscopy. Both the mononuclear and the dinuclear complexes promote the CO/styrene copolymerization, yielding the corresponding polyketone with a fully or a predominantly isotactic microstructure, depending on the reaction medium. The nature of the anion present in the palladium precatalysts affects the polyketone stereochemistry. MALDI-TOF analysis of the copolymers synthesized reveals the presence of p-hydroxyphenolic end-groups, thus confirming and explaining the role of 1,4-hydroquinone as a molecular weight regulator. PMID- 16047395 TI - Preparation, characterization, and application of multistimuli-responsive microspheres with fluorescence-labeled magnetic cores and thermoresponsive shells. AB - Novel functional microspheres with multistimuli-responsive properties have been prepared and characterized. The as-prepared microspheres respond to an external magnetic field, environmental temperature, and ultraviolet radiation. The in vitro drug-loading efficiency and drug-release behavior of these microspheres demonstrated that they could be used as drug carriers for drug controlled release. The results of in vivo distribution investigations of these microspheres showed that they exhibit a high magnetic-targeting effect, which holds promise for applications in various fields such as magnetic drug targeting and tissue labeling, among others. PMID- 16047396 TI - Opinion on algal toxins. PMID- 16047398 TI - An evaluation, comparison, and accurate benchmarking of several publicly available MS/MS search algorithms: sensitivity and specificity analysis. AB - MS/MS and associated database search algorithms are essential proteomic tools for identifying peptides. Due to their widespread use, it is now time to perform a systematic analysis of the various algorithms currently in use. Using blood specimens used in the HUPO Plasma Proteome Project, we have evaluated five search algorithms with respect to their sensitivity and specificity, and have also accurately benchmarked them based on specified false-positive (FP) rates. Spectrum Mill and SEQUEST performed well in terms of sensitivity, but were inferior to MASCOT, X!Tandem, and Sonar in terms of specificity. Overall, MASCOT, a probabilistic search algorithm, correctly identified most peptides based on a specified FP rate. The rescoring algorithm, PeptideProphet, enhanced the overall performance of the SEQUEST algorithm, as well as provided predictable FP error rates. Ideally, score thresholds should be calculated for each peptide spectrum or minimally, derived from a reversed-sequence search as demonstrated in this study based on a validated data set. The availability of open-source search algorithms, such as X!Tandem, makes it feasible to further improve the validation process (manual or automatic) on the basis of "consensus scoring", i.e., the use of multiple (at least two) search algorithms to reduce the number of FPs. complement. PMID- 16047399 TI - By the way, doctor. I have irritable bowel syndrome, with frequent bloating and constipation. My doctor prescribed a six-week course of Zelnorm, which really helped. Is it safe to take long-term? PMID- 16047400 TI - Treatment-experienced patients set high standards. PMID- 16047401 TI - Letter regarding article by Nasir et al, "Electrocardiographic features of arrhythmogenic right ventricular dysplasia/cardiomyopathy according to disease severity: a need to broaden diagnostic criteria". PMID- 16047402 TI - The role of the junior surgical trainee in ward rounds. PMID- 16047403 TI - Bibliography. Current world literature. Paediatric urology. PMID- 16047404 TI - Retention of the 4-pro-R hydrogen atom of mevalonate at C-2,2' of bacterioruberin in Halobacterium halobium. PMID- 16047405 TI - Proceedings of the 4th Amino Acid Assessment Workshop. October 28-29, 2004. Kobe, Japan. PMID- 16047406 TI - Smoke without fire: the complex effects of cigarette smoking on thrombolytic therapy for acute ischemic stroke. PMID- 16047407 TI - One city, one world. PMID- 16047408 TI - Tipranavir (Aptivus): approval cautiously recommended. AB - A new protease inhibitor active against most HIV that is resistant to other protease inhibitors is likely to be approved in the U.S. soon, after an 11-3 vote of an advisory committee. PMID- 16047409 TI - ADAP status now: what you can do. AB - This short review of the current state of the AIDS Drug Assistance Program tells how you can get involved. PMID- 16047410 TI - Lexiva: blood levels not lowered when taken simultaneously with Nexium. AB - A study suggested that the two drugs might be used together, without lowering the blood level of Lexiva. But the timing of the doses may be critical. PMID- 16047411 TI - Medical innovation prize fund: new idea in drug development. AB - This proposal, now introduced in Congress as HR 417, would replace current drug marketing with system better designed to reward effective innovation. All drugs would be treated as generics immediately when approved by the FDA, and patent holders would be rewarded from a $60 billion a year award fund for innovations that actually led to better health. PMID- 16047412 TI - Assessment of genetic diversity among 16 promising cultivars of ginger using cytological and molecular markers. AB - Ginger (Zingiber officinale Roscoe) is an economically important plant, valued all over the world. The existing variation among 16 promising cultivars as observed through differential rhizome yield (181.9 to 477.3 g) was proved to have a genetic basis using different genetic markers such as karyotype, 4C nuclear DNA content and random amplified polymorphic DNA (RAPD). The karyotypic analysis revealed a differential distribution of A, B, C, D and E type of chromosomes among different cultivars as represented by different karyotype formulas. A significant variation of 4C DNA content was recorded in ginger at an intraspecific level with values ranging from 17.1 to 24.3 pg. RAPD analysis revealed a differential polymorphism of DNA showing a number of polymorphic bands ranging from 26 to 70 among 16 cultivars. The RAPD primers OPC02, OPA02, OPD20 and OPN06 showing strong resolving power were able to distinguish all 16 cultivars. The extent of genetic diversity among these cultivars was computed through parameters of gene diversity, sum of allele numbers per locus and Shannon's information indices. Cluster analysis, Nei's genetic similarity and genetic distances, distribution of cultivars into special distance classes and principal coordinate analysis and the analysis of molecular variance suggested a conspicuous genetic diversity among different cultivars studied. The genetic variation thus detected among promising cultivars of ginger has significance for ginger improvement programs. PMID- 16047413 TI - A simple method to obtain essential oils from Salvia triloba L. and Laurus nobilis L. by using microwave-assisted hydrodistillation. AB - A microwave-assisted hydrodistillation protocol was modified to extract essential oils from leaves of Salvia triloba L. and Laurus nobilis L. The essential oils of these plants are generally obtained by hydrodistillation or steam distillation. The volatile compounds obtained by microwave-assisted hydrodistillation and hydrodistillation methods were analyzed by GC and GC/MS. Both distillation methods and analytical results were compared. 1,8-Cineole (46.8-54.2%) was the main component in the leaf oils of both samples. Although the distillation was accomplished in a shorter time, oil yields and 1,8-cineole contents were slightly higher in the microwave-assisted hydrodistillation compared to usual hydrodistillation. Microwave-assisted hydrodistillation appears to be an effective method for the production of essential oils. PMID- 16047414 TI - Klebsiella pneumoniae type 3 fimbria-mediated immunity to infection in the murine model of respiratory disease. AB - Type 3 fimbriae are expressed by most strains of Klebsiella pneumoniae and facilitate adherence to the basement membrane of human respiratory tissues. The ability of these appendages to stimulate a protective immune response in vivo has not been investigated. A murine model of acute pneumonia was used to determine whether the production of type 3 fimbria-specific antibodies correlated with protection against infection by K. pneumoniae. Purified fimbriae from several strains were used to immunize mice prior to challenge with a virulent strain. The immunized mice produced high titers of specific antibody and this was associated with protection against challenge with a low dose of bacteria that was lethal in unimmunized animals. However, challenge with a high number of bacteria resulted in no protection against infection. PMID- 16047415 TI - Extracellular matrix-driven alveolar epithelial cell differentiation in vitro. AB - During homeostasis and in response to injury, alveolar type II (AT2) cells serve as progenitor cells to proliferate, migrate, differentiate, and re-establish both alveolar type I (AT1) and AT2 cells into a functional alveolar epithelium. To understand specific changes in cell differentiation, we monitored morphological characteristics and cell-specific protein markers over time for isolated rat AT2 cells cultured on combinations of collagen, fibronectin and/or laminin-5 (Ln5). For all matrices tested, cultured AT2 cells displayed reduced expression of AT2 cell-specific markers from days 1 to 4 and increased expression of AT1-specific markers by day 3, with continued expression until at least day 5. Over days 5 to 7 in culture, cells took on an AT1-like phenotype (on collagen/fibronectin alone; collagen alone; or Ln5 alone), an AT2-like phenotype (on collagen/fibronectin/Ln5; or collagen/Ln5), or both AT1-like and AT2-like phenotypes (on collagen/fibronectin matrix with a subsaturating amount of Ln5). Cells transferred between matrices at day 4 of culture retained the ability to alter day 7 phenotype. We conclude that in vitro, (1) AT2 cells exhibited phenotype plasticity that included an intermediate cell type with both AT1 and AT2 cell characteristics independent of day 7 phenotype; (2) both collagen and Ln5 were needed to promote the development of an AT2-like phenotype at day 7; and (3) components of the extracellular matrix (ECM) contribute to phenotypic switching of alveolar cells in culture. The described tissue culture models provide accessible models for studying changes in alveolar epithelial cell physiology from AT2 cell progenitors to the establishment of alveolar epithelial monolayers that represent AT1-like, AT2-like, or a mix of AT1- and AT2-like cells. PMID- 16047416 TI - Add-back of allodepleted donor T cells to improve immune reconstitution after haplo-identical stem cell transplantation. AB - Poor immune reconstitution after haplo-identical stem cell transplantation results in high mortality from viral infections and relapse. One approach to overcome this problem is to deplete alloreactive cells selectively by deleting T cells activated by recipient stimulators, using an immunotoxin directed against the activation marker CD25. However, the degree of depletion of alloreactive cells is variable following stimulation with recipient PBMC, and this can result in GvHD. We have shown that using recipient EBV-transformed LCL as stimulators to activate donor alloreactive T cells results in more consistent depletion of in vitro alloreactivity while preserving T-cell responses to viral and potential myeloid tumor Ag. Based on these data, we have embarked on a phase I clinical dose escalation study of add-back of allo-LCL-depleted donor T cells in the haplo identical setting, to determine if the allodepletion we achieve to allow infusion of sufficient T cells to restore useful antiviral/anti-leukemic responses without causing GvHD. Fifteen patients have so far been treated. The incidence of significant acute or chronic GvHD has been low (2/15), as has mortality from infection (1/15). Preliminary data show accelerated immune reconstitution in dose level 2 patients. Infused allodepleted donor T cells appear able to expand significantly in the face of viral reactivations, and doses as low as 3 x 10(5)/kg may be sufficient to confer useful antiviral immunity in this setting. At a median follow-up of 19.5 months, nine of 15 patients are alive and disease free. Five patients have relapsed, all of whom have died. PMID- 16047417 TI - [Efficiency in a disease management program in asthma]. PMID- 16047418 TI - After Bristol: the healthcare of young children and the law. AB - This paper considers the written statements provided to the Bristol Inquiry by parents whose children underwent cardiac surgery at the Bristol Royal Infirmary between 1984 and 1995, seeking to learn from their experiences, opinions, feelings and expectations. The law regulating the relationship between healthcare professional, parent and child is considered in light of these accounts. The limitations of the existing law are such that a new legal framework is required which fosters the relationship between healthcare professional, parent and child, supporting them in the shared endeavour of caring for the child. Of central importance within this new framework would be recognition of each child as a distinct individual and of the expertise which parents can contribute to the care of their child. PMID- 16047419 TI - In-vitro fertilization, frozen embryos, and the right to privacy--are mandatory donation laws constitutional? PMID- 16047420 TI - Right to refuse life sustaining medical treatment and the noncompetent nonterminally ill patient: an analysis of abridgment and anarchy. PMID- 16047421 TI - Antinomy: the use, rights and regulation of laboratory animals. PMID- 16047422 TI - Reproductive biotechnology: an animal scientist's perspective. AB - This paper questions the extent of the "cultural mandate" given humankind in Genesis. How far does the authority granted to us extend? With the emergence of new biotechnologies that deal with reproduction come new questions of what science should and should not do. These questions are of added concern because techniques for human medicine are first developed in animal laboratories. It is suggested that new technologies should be developed for use in laboratory and domestic animals since they can benefit humankind, but that more restraint and prayerful consideration of each technological advance should be made before its application to human medicine. PMID- 16047423 TI - The right to die. PMID- 16047424 TI - Victorian secret: pro-life victories in 19th-century America. PMID- 16047425 TI - Effects of readability on the impact and recall of written informed consent material. AB - Participants (49 women, 23 men) rated a hypothetical psychologist after (a) receiving either no informed consent information, a highly readable consent form, or a less readable form; and (b) reading a transcript of a therapy session containing either positive or negative self-involving statements. Informed consent forms had no effect on ratings; there was a trend for participants to recall less of the less readable form. Participants rated the therapist more favorably when he used positive self-involving statements. In a second study, participants (62 women, 28 men) received 1 of the 3 levels of informed consent information and a transcript with either no mention of informed consent or a brief mention at the end. Men who received the less readable consent form had poorer first impressions of the therapist and recalled less of the form. The mention of informed consent in the transcript had no effect. PMID- 16047426 TI - Financial penalties for failing to honor patient wishes to refuse treatment. PMID- 16047427 TI - Ethics and the professional practice of psychologists: the role of virtues and principles. AB - We evaluate the potential relevance of virtue ethics to the training and practice of professional psychologists, and we contrast them with principle ethics. Typically, principles are used to facilitate the selection of socially and historically acceptable answers to the question "What shall I do?" when confronted by ethical dilemmas. Virtue ethics, however, generally focus on the question "Who shall I be?" Strengths and weaknesses of each approach are presented. The impact of each is discussed with respect to informed consent and the therapeutic construct "genuineness." We conclude that virtue ethics are an essential component of responsible ethical training and practice. PMID- 16047428 TI - Do written consent forms influence clients' first impressions of therapists? AB - Students (24 men, 105 women) rated a hypothetical psychologist after reading some combination of (a) a question sheet (Handelsman & Galvin, 1988), (b) a form designed to conform to a new Colorado law, and (c) a general brochure about psychologists. The legal form increased the likelihood of recommending "Dr. Doe" to a friend; when the brochure was included, the form also increased judgments of experience. The question sheet increased recommendations, judgments of likeability and trustworthiness, and (when the brochure was absent) the likelihood of going to Dr. Doe oneself. In a second study, students (44 men, 93 women) received the legal disclosure form; some also received the question sheet. They were told that Dr. Doe had 0, 9, or 19 years of experience. The question sheet increased ratings of trustworthiness and likelihood of going oneself. Therapists with at least 9 years of experience were recommended more. PMID- 16047429 TI - Sexual misconduct in psychotherapy: reactions to a consumer-oriented brochure. AB - Reactions of subjects to a brochure addressing the topic of sexual intimacy in psychotherapy was assessed. Fifty-four psychotherapy clients and 52 licensed psychologists responded favorably to the brochure, with the majority indicating that the brochure should be made available before therapy begins or during the first session. Potential clients (120 college students) read either the brochure on sexual misconduct or a control brochure containing general information about psychology. Subjects who read the sexual misconduct brochure showed increased ability to understand what is appropriate and inappropriate therapist behavior, see unwanted touch as nontherapeutic, regard a therapist talking a lot about his or her own sex life as inappropriate, view sex in the therapeutic relationship as inappropriate, and intend to behave assertively within the session if their therapist's behavior should make them feel uncomfortable. Subjects who read the control brochure showed no such changes. PMID- 16047430 TI - Pro-child/pro-choice: an exercise in doublethink? PMID- 16047431 TI - Do animal rights entail moral nihilism? PMID- 16047432 TI - Drug-assisted intubation in the prehospital setting. PMID- 16047433 TI - Empowerment: reformulation of a non-Rogerian concept. PMID- 16047434 TI - Health Savings Accounts as a tool for market change. PMID- 16047435 TI - Guideline at-a-glance. AHA, ASA revisit stroke guidelines, emphasize education of patient. PMID- 16047436 TI - Ethical implications of for-profit corporate sponsorship of research. AB - Academic institutions and commercial companies both recognize the need for the United States to enhance its economic competitiveness worldwide. Improved trading status is linked, in part, to education and to research and development (R&D) as cooperative efforts between nonprofit colleges and universities and for-profit corporations. As more and more colleges and universities seek for-profit corporate sponsors for their research programs, their collaboration may be complicated by several ethical issues. Most of these issues arise because of the differences between academic and corporate missions. This paper emphasizes that these differences are not irreconcilable; indeed, there are many successful research partnerships between academia and for-profit corporations. With a solid understanding of each partner's mission, productive corporate sponsorship of academic research is possible. PMID- 16047437 TI - Practice management. PMID- 16047438 TI - The ethics of reproductive cloning. PMID- 16047439 TI - Ethics of nanobiotechnology at the frontline. PMID- 16047440 TI - The role of contract principles in determining the validity of surrogacy contracts. PMID- 16047441 TI - Rights of animals, perceptions of science, and political activism: profile of American animal rights activities. AB - This article reports original research examining characteristics of the active followers of the American animal rights movement. Typical respondents were Caucasian, highly educated urban professional women approximately thirty years old with a median income of $33,000 (1989). Most activists think of themselves as Democrats or as Independents, and have moderate to liberal political views. They were often suspicious of science and made no distinction between basic and applied science, or public versus private animal-based research. The research suggests that animal rights activism is part of a symbolic manifestation of egalitarian social and political views concerning scientific and technological change. PMID- 16047442 TI - Donohue lecture series: "Roe v. Wade: past and future". PMID- 16047443 TI - Can fetal tissue transplantation be done legally? PMID- 16047444 TI - The validity of legislative restrictions on abortion under the Oregon constitution. PMID- 16047445 TI - Biology and the future: a doctrinal agenda. PMID- 16047446 TI - Why is health care special? PMID- 16047447 TI - Frozen embryos: towards an equitable solution. PMID- 16047448 TI - The disfranchisement of fertile women in clinical trials: the legal ramifications of and solutions for rectifying the knowledge gap. PMID- 16047449 TI - The Oregon Basic Health Services Act: a model for state reform? PMID- 16047450 TI - Cruel and unusual punishment in the provision of prison medical care: challenging the deliberate indifference standard. PMID- 16047451 TI - The pariah patient: the lack of funding for mental health care. PMID- 16047452 TI - Abortion 1990s: contemporary issues and the activist court. PMID- 16047453 TI - Health care representative consent legislation. PMID- 16047454 TI - Jones v. Gerhardstein: the involuntarily committed mental patient's right to refuse treatment with psychotropic drugs. AB - The question of whether an involuntary committed mental patient has a fundamental right to refuse treatment with psychotropic drugs continues to be a subject of much debate. Over the past twenty-five years, psychotropic drugs have become the most common form of treatment for the mentally ill. For many patients, these drugs provide substantial benefits; for others, however, they produce severe, sometimes debilitating, side effects. Because of the possibility of serious harm to the patient and because of the potential for abuse of drug treatment by psychiatric staffs, the mental health bar generally has argued for increased procedural protection for mental patients. In Jones v. Gerhardstein, the Wisconsin Supreme Court responded to these concerns by requiring that a judicial hearing be held on the issue of a patient's competency to refuse treatment before the attending physician may administer medication without the patient's consent. This Note discusses the controversy between the legal and medical communities over treatment refusal by mentally ill patients in light of the impact of the Jones decision on institutional practice and on refusing patients. The author argues that the strictly rights-based analysis used by the Jones court has done little to benefit involuntarily committed mental patients. The author suggests alternative ways of approaching treatment refusal that might be more responsive to the distinctive needs of the mentally ill. PMID- 16047455 TI - The supply chain and clinical quality. PMID- 16047456 TI - The rules versus similarity distinction. AB - The distinction between rules and similarity is central to our understanding of much of cognitive psychology. Two aspects of existing research have motivated the present work. First, in different cognitive psychology areas we typically see different conceptions of rules and similarity; for example, rules in language appear to be of a different kind compared to rules in categorization. Second, rules processes are typically modeled as separate from similarity ones; for example, in a learning experiment, rules and similarity influences would be described on the basis of separate models. In the present article, I assume that the rules versus similarity distinction can be understood in the same way in learning, reasoning, categorization, and language, and that a unified model for rules and similarity is appropriate. A rules process is considered to be a similarity one where only a single or a small subset of an object's properties are involved. Hence, rules and overall similarity operations are extremes in a single continuum of similarity operations. It is argued that this viewpoint allows adequate coverage of theory and empirical findings in learning, reasoning, categorization, and language, and also a reassessment of the objectives in research on rules versus similarity. PMID- 16047457 TI - A refined model of sleep and the time course of memory formation. AB - Research in the neurosciences continues to provide evidence that sleep plays a role in the processes of learning and memory. There is less of a consensus, however, regarding the precise stages of memory development during which sleep is considered a requirement, simply favorable, or not important. This article begins with an overview of recent studies regarding sleep and learning, predominantly in the procedural memory domain, and is measured against our current understanding of the mechanisms that govern memory formation. Based on these considerations, I offer a new neurocognitive framework of procedural learning, consisting first of acquisition, followed by two specific stages of consolidation, one involving a process of stabilization, the other involving enhancement, whereby delayed learning occurs. Psychophysiological evidence indicates that initial acquisition does not rely fundamentally on sleep. This also appears to be true for the stabilization phase of consolidation, with durable representations, resistant to interference, clearly developing in a successful manner during time awake (or just time, per se). In contrast, the consolidation stage, resulting in additional/enhanced learning in the absence of further rehearsal, does appear to rely on the process of sleep, with evidence for specific sleep-stage dependencies across the procedural domain. Evaluations at a molecular, cellular, and systems level currently offer several sleep specific candidates that could play a role in sleep-dependent learning. These include the upregulation of select plasticity associated genes, increased protein synthesis, changes in neurotransmitter concentration, and specific electrical events in neuronal networks that modulate synaptic potentiation. PMID- 16047458 TI - Appropriate use of intravenous proton pump inhibitors in the management of bleeding peptic ulcer. AB - Rebleeding from peptic ulcers is a major unsolved problem in the management of acute upper gastrointestinal bleeding. Our goal was to review what is known and what remains to be learned about the effectiveness of antisecretory therapy for acute upper gastrointestinal bleeding. We reviewed the data regarding the effectiveness of endoscopic therapy, the prediction of those at increased risk for rebleeding, and the effectiveness of antisecretory drug therapy in preventing rebleeding with or without endoscopic hemostasis. Proton pump inhibitor therapy without endoscopic hemostasis is ineffective clinically for stopping bleeding or preventing rebleeding. Endoscopic hemostasis remains the cornerstone of therapy. The data are consistent with the notion that reliable maintenance of the intragastric pH at > or = 6 after endoscopic hemostasis is associated with the lowest rebleeding rates. H2-receptor antagonists are ineffective for achieving this goal. Intermittent bolus and oral administration of proton pump inhibitors are equivalent and fail to achieve this goal, which can only be accomplished by bolus administration of a proton pump inhibitor (e.g., 80 mg) followed by a constant infusion (e.g., 8 mg/hr). Whether the combination of endoscopic hemostasis and pH control is equal or superior to selected second-look endoscopy is unknown. A treatment algorithm is suggested. PMID- 16047459 TI - The efficacy of high- and low-dose intravenous omeprazole in preventing rebleeding for patients with bleeding peptic ulcers and comorbid illnesses. AB - This study sought to determine if high-dose omeprazole infusion could improve the control of rebleeding in patients with comorbid illnesses and bleeding peptic ulcers. After achieving hemostasis by endoscopy, 105 patients were randomized into high-dose (n = 52) and low-dose (n = 53) groups, receiving 200 and 80 mg/day omeprazole, respectively, as a continuous infusion for 3 days. Thereafter, oral omeprazole, 20 mg/day, was given. The cumulative rebleeding rates comparatively rose in both groups (high-dose vs. low-dose group), beginning on day 3 (15.4% vs. 11.3%), day 7 (19.6% vs. 20%), and day 14 (32.7% vs. 28.9%), until day 28 (35.4% vs. 33.3%), and were not significantly different between the two groups (P > 0.50). Multiple logistic regression confirmed that a serum albumin level <3 g/dL was an independent factor associated with rebleeding (P = 0.002). For patients with comorbidities, 3-day omeprazole infusion, despite increasing the daily dose from 80 to 200 mg, was not adequate to control peptic ulcer rebleeding. PMID- 16047460 TI - Acid-suppressive efficacy of a reduced dosage of rabeprazole: comparison of 10 mg twice daily rabeprazole with 20 mg twice daily rabeprazole, 30 mg twice daily lansoprazole, and 20 mg twice daily omeprazole by 24-hr intragastric pH-metry. AB - Rabeprazole achieves more potent acid suppression than other proton pump inhibitors. Therefore it is administered at reduced as well as high dosages in eradication therapy for Helicobacter pylori; however, there is incomplete assessment of the efficacy of a reduced dosage of rabeprazole as might be employed in therapy. In this study, we evaluated acid-suppressive efficacy of a reduced dosage of rabeprazole on day 7 by 24-hr pH-metry in 10 healthy male cytochrome P-450 2C19 extensive metabolizers without Helicobacterpylori infection and compared the results with those of high dosages of rabeprazole, lansoprazole, and omeprazole. Median intragastric pH value, pH >3 holding time ratio (pH>3HT), pH>4HT, pH>5HT, pH>6HT, and pH>7HT for 24 hr with rabeprazole, 10 mg twice daily, were not significantly different from those of rabeprazole, 20 mg twice daily, lansoprazole, 30 mg twice daily, and omeprazole, 20 mg twice daily. In conclusion, for acid-suppressive efficacy, a reduced dosage of rabeprazole is comparable to high dosages of rabeprazole, lansoprazole, and omeprazole. PMID- 16047461 TI - Influence of 1-week Helicobacter pylori eradication therapy with rabeprazole, clarithromycin, and metronidazole on 13C-aminopyrine breath test. AB - Helicobacter pylori eradication therapy is commonly prescribed in the general population. Treatment consists of drugs that are mainly metabolized by the liver cytochrome P-450 (CYP) enzymatic pool. Most H. pylori-infected patients often take drugs for comorbid illnesses, therefore increasing the potential for drug drug interactions. We aimed to evaluate the interactions of rabeprazole, clarithromycin, and metronidazole 1-week H. pylori eradication therapy with CYP dependent liver metabolic function in clinical practice. Ten patients referred to our unit for H. pylori infection underwent 1-week eradication therapy with rabeprazole (20 mg, b.i.d.), clarithromycin (500 mg, b.i.d.), and metronidazole (500 mg, b.i.d.). We chose the 13C-aminopyrine breath test (13C-ABT) to evaluate CYP-dependent liver function since it is noninvasive and nonharmful. All patients underwent 13C-ABT at three time points: before therapy (to), at the end of therapy (t8), and after 1 month of follow-up (t38). Mean 13C-ABT dose/hr (t0 = 14.0 +/- 5.4, t8 = 13.5 +/- 4.0, t38 = 16.1 +/- 5.6) as well as 13C-ABT cumulative dose (t0 = 2.4 +/- 1.1, t8 = 2.4 +/- 0.8, t38 = 2.6 +/- 1.0) were not statistically different at the three time points of the study. These results did not seem to be influenced by drugs being administered concomitantly. In everyday clinical practice rabeprazole-based H. pylori eradication therapy does not seem to display any significant interactions with CYP-dependent liver function, even in patients on multiple drugs. PMID- 16047462 TI - The relationship of ABO blood group, age, gender, smoking, and Helicobacter pylori infection. AB - It is well known that blood group antigens are related to the development of peptic ulcer and gastric carcinoma. This study sought to determine the relationship between H. pylori and ABO/Rhesus blood groups, age, gender, and smoking. Patients (335 women and 205 men; mean age, 51.68 +/- 15.0 years; range, 18-85 years) who attended our outpatient clinic were enrolled in the study. All patients were randomly selected in each age group. Demographic data recorded for each patient included age, gender, and tobacco use. Blood samples were tested for H. pylori antibodies, and ABO/Rhesus blood group antigen typing was performed. Serum antibodies were tested against H. pylori infection. Prevalences of all blood groups were O (29.2%), A (38.2%), B (17.8%), and AB (14.8%). As expected from previous studies, we found that seropositivity for H. pylori increased with age. H. pylori Ig G antibody positivity was detected in 185 of 335 women (60.6%), compared with 88 of 205 men (42.9%), a statistically significant difference (P < 0.05). H. pylori Ig G antibody positivity was detected in 206 of 379 nonsmokers (54.3%) compared with 67 of 161 smokers (41.6%), a statistically significant difference (P < 0.05). Patients in blood groups A and O were more prone to H. pylori infection than were patients in other blood groups (P < 0.05), and patients in the AB blood group were less prone to H. pylori infection compared with patients in other blood groups (P < 0.05). The results of this study demonstrate that H. pylori infection can be related to ABO blood group, age, gender, and smoking. PMID- 16047463 TI - Rescue of Helicobacter pylori-induced cytotoxicity by red ginseng. AB - Helicobacter pylori has been known to provoke gastric inflammation, ulceration, and DNA damage, based on which WHO defined H. pylori as a class I carcinogen. Although ginseng, the root of Panax ginseng C.A. Meyer, has been reported to possess antiadhesion or antimicrobial activity against H. pylori, in this study, we examined the protective effect of red ginseng extracts (RGE) against H. pylori induced cytotoxicity and DNA damage. RGE significantly attenuated both H. pylori induced DNA damage assessed by comet assay and apoptosis measured by DNA fragmentation. Inactivation of ERK1/2 signaling and attenuation of caspase-3 activation and PARP cleavage were revealed with RGE against H. pylori infection. RGE decreased H. pylori-stimulated IL-8 gene expression, which resulted from the transcriptional regression of NF-kappaB. In conclusion, RGE showed significant gastroprotective effects against H. pylori-associated gastric mucosal cell damage, suggesting that red ginseng could be used as a medicinal phytonutrient against H. pylori infection. PMID- 16047464 TI - Does eradication of Helicobacter pylori infection help normalize serum lipid and CRP levels? AB - It is still unclear whether Helicobacter pylori infection is associated with risk factors for coronary artery disease. The aim of this study was to determine whether eradication of H. pylori infection affects serum lipid levels and C reactive protein (CRP) levels. Seventy-eight patients who had H. pylori antigen positivity in their stools were enrolled. Clarithromycin, 1 g/day, amoxicillin, 2 g/day, and omeprazole, 40 mg/day, were given for 14 days. Serum total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglyceride (TG), and CRP were measured at baseline and 8 weeks after therapy. According to H. pylori stool antigen study after 8 weeks, individuals in whom H. pylori was eradicated were recruited as group A and those in whom H. pylori was not eradicated formed group B. Group A comprised 57 patients, and group B 21 patients. Patients in group A comprised 32 women and 25 men and their ages ranged from 35 to 59 years. Patients in group B included 13 women and 8 men, aged 32-61 years. No significant difference in LDL, TC, or TG serum levels were found between group A and group B. Although CRP and HDL serum levels were found to be the same before and after treatment in group B, CRP levels were found to decrease and HDL levels to increase significantly in group A (P < 0.05). We conclude that H. pylori infection may affect lipid metabolism in a way that could increase the risk of atherosclerosis. Thus H. pylori infection is an independent risk factor for coronary artery disease. PMID- 16047465 TI - Effects of colesevelam HC1 on sterol and bile acid excretion in patients with type IIa hypercholesterolemia. AB - Colesevelam HC1 is a potent bile acid-binding polymer. This study's aim was to determine effects of colesevelam HCl on sterol and bile acid excretion in patients with type IIa hypercholesterolemia. Twenty-four patients (low-density lipoprotein cholesterol, 130 to 220 mg/dL) enrolled in an open-label, parallel design study, entered an American Heart Association/National Cholesterol Education Program diet for 6 weeks and were randomized to colesevelam HCl, 2.3 or 3.8 g/day for 4 weeks. In an apparent dose-related manner, respective mean serum concentrations HCl of low-density lipoprotein cholesterol decreased by 10% (P < 0.01) and 13% (P = 0.05), mean total cholesterol levels decreased by 4.9% (P = 0.05) and 6.1% (P = 0.09), and total fecal bile acid excretion showed median changes of +324% (P < 0.05) and +316% (P < 0.05). Colesevelam HCl did not affect fecal neutral sterol or fecal fatty acid excretion; however, 24-hr urinary mevalonic acid levels significantly increased in both treatment groups (P < 0.05). The cholesterol-lowering action of colesevelam HCl appears to be mediated through increased bile acid excretion. PMID- 16047466 TI - Intestinal and systemic effects of oral insulin supplementation in rats after weaning. AB - Oral insulin has intestinal trophic effects in suckling animals. In mice, lower glucose and lipid levels may be seen when oral insulin is given after the weaning period. The purpose of the present study is to examine local and systemic effects of oral insulin supplementation in rats in the postweaning period. Immediately after weaning, Sprague-Dawley rats received either drinking water (controls) or oral insulin in their drinking water (1 U/ml) for either 1 week or 6 weeks. Intestinal mucosal parameters (bowel and mucosal weight, mucosal DNA and protein) and histological changes were examined in all study groups. Glucose levels were monitored weekly, and at the end of the study, blood levels of glucose, lipids, and insulin were measured in the fasting state. After 1 week of insulin supplementation, mucosal weight in duodenum and jejunum as well as jejunal DNA content were significantly higher in insulin-supplemented rats compared to controls. Duodenal circumference and villus height in jejunum were significantly higher in insulin-supplemented rats compared to controls on both day 7 and day 42. Total cholesterol levels were significantly lower in the study group compared with the controls. We conclude that oral insulin supplementation exerts intestinal trophic effects, as well as systemic effects in the postweaning period in rats. PMID- 16047467 TI - Subjective health complaints and modern health worries in patients with subjective food hypersensitivity. AB - Perceived food hypersensitivity is much more common than food allergy as medically verified. Unexplained symptoms and wrong attribution are typical in subjective health complaints. We hypothesize that subjective health complaints and worries are abnormally prevalent among patients with subjective food hypersensitivity. Forty-six patients with subjective food hypersensitivity and two control groups, one formed by 50 health care workers and one by 70 sex- and age-matched volunteers from the general population, were included in our study. All filled in two questionnaires: Subjective Health Complaints Inventory and Modern Health Worries Scale. None of the patients had IgE-mediated food allergy. The patients scored significantly higher than the controls on sum scores for four domains of subjective health complaints, including gastrointestinal complaints (P < 0.001), musculoskeletal complaints (P < 0.01), "pseudoneurology" (P < 0.001), and allergy (P < 0.001). Sum scores on modern health worries did not differ significantly between groups. The results support our hypothesis of an association between subjective food hypersensitivity and subjective health complaints, corroborating the view that, in the absence of food allergy, the conditions are sharing pathogenetic mechanisms. PMID- 16047468 TI - Enhanced human beta-defensin-2 (hBD-2) expression by corticosteroids is independent of NF-kappaB in colonic epithelial cells (CaCo2). AB - Beta-defensins are small cationic peptides with antimicrobial properties that contribute to innate host defense. Unlike human beta-defensin-1 (hBD-1), which is produced constitutively, human beta-defensin-2 (hBD-2) is expressed after adequate stimulation by cytokines and/or bacterial endotoxins in epithelial tissue and mononuclear phagocytes but may be deficient in patients with Crohn's disease. To further elucidate the role of the intestinal epithelium in antimicrobial host defense, gene regulation of hBD-2 and the interaction with NF kappaB were analyzed in a cell culture model. Human colonic epithelial cells (CaCo2) were stimulated by pro-inflammatory cytokines (IL-1beta, TNF-alpha, IF gamma) to induce hBD-2 mRNA transcription. Interactions with NF-kappaB were analyzed using specific inhibitors (sulfasalazine, gliotoxine, dexamethasone) at different concentrations. Defensin mRNA expression was quantified by competitive RT-PCR and antibacterial capacity of supernatants was determined by an antimicrobial assay. HBD-2 mRNA transcription and antimicrobial activity of CaCo2 cells were induced by stimulation with pro-inflammatory cytokines. Induction was not inhibited by sulfasalazine or gliotoxine, whereas dexamethasone further enhanced both gene transcription and antimicrobial capacity. The lack of inhibition of induced hBD-2 expression by specific NF-kappaB antagonists suggests an additional pathway of activation, independent of NF-kappaB. The induction of hBD-2 expression in cytokine-stimulated CaCo2 cells by corticosteroids indicates further immunomodulatory ability of steroid hormones not yet understood. PMID- 16047469 TI - Perianal fistulas in Crohn's disease are predominantly colonized by skin flora: implications for antibiotic treatment? AB - In this study we aimed to determine the microorganisms found in perianal fistulas in Crohn's disease and whether treatment with ciprofloxacin affects these microorganisms. Thirteen patients (males/females, 7/6; median age, 34 years; range, 18-61 years) with fistulas were treated with infliximab, 5 mg/kg intravenously, at weeks 6, 8, and 12 and randomized to double-blind treatment with ciprofloxacin, 500 mg bd (n = 6), or placebo (n = 7) for 12 weeks. Samples were taken at baseline and at weeks 6 and 18. In the ciprofloxacin group 10 different genera of microorganisms were identified, while 13 genera could be identified in the placebo group. Gram-negative enteric floras were present in a small minority. The genera found in patients with perianal fistulas were predominantly gram-positive microorganisms. Therefore, antimicrobial treatment should be directed toward these microorganisms. PMID- 16047470 TI - Electroacupuncture reduces rectal distension-induced blood pressure changes in conscious dogs. AB - It has been shown that acupuncture relieves symptoms of abdominal pain and bloating in patients with irritable bowel syndrome (IBS). However, the mechanism of beneficial effects of acupuncture still remains unproven. The aim of the present study was to investigate the mechanisms of the antinociceptive effects of acupuncture in conscious dogs. We evaluated the increase in mean arterial blood pressure (MAP) caused by rectal distension as an index of visceral pain. Electroacupuncture (EA; 10 Hz) at ST-36 (lower leg), but not at BL-21 (back), significantly reduced the increase in MAP in response to rectal distension (30 and 40 cm3). The antinociceptive effect of EA at ST-36 was abolished by pretreatment with naloxone (a central and peripheral opioid receptor antagonist) but not by naloxone methiodide (a peripheral opioid receptor antagonist). These results suggest that EA at ST-36 may reduce visceral pain via central opioid pathway. Acupuncture may be useful to treat visceral hypersensitivity in IBS patients. PMID- 16047471 TI - Clostridium difficile toxin A induces intestinal epithelial cell apoptosis and damage: role of Gln and Ala-Gln in toxin A effects. AB - The aim of this study was to investigate the effect of Clostridium difficile toxin A (TxA) on intestinal epithelial cell migration, apoptosis, and transepithelial resistance and to evaluate the effect of glutamine (Gln) and its stable derivative, alanyl-glutamine (Ala-Gln), on TxA-induced damage. Migration was measured in rat intestinal epithelial cells (IEC-6) 6 and 24 hr after a razor scrape of the cell monolayer. Cell proliferation was indirectly measured utilizing the tetrazolium salt WST-1. The cells were incubated with TxA (1-100 ng/ml) in medium without Gln or medium containing Gln or Ala-Gln (1-30 mM). Apoptosis was quantified in IEC-6 cells using annexin V assay. Transepithelial resistance was measured using an epithelial voltohmmeter across T84 cells seeded on a transwell filter. TxA-induced a dose-dependent reduction of migration and also caused dose and time-dependent apoptosis in IEC-6 cells. Gln and Aln-Gln significantly enhanced IEC-6 cell migration and proliferation. Gln and Ala-Gln also prevented the inhibition of migration, apoptosis, and the initial drop in transepithelial resistance induced by TxA. In conclusion, both peptides reduced toxin-induced epithelial damage and thus might play an adjunctive role in C. difficile-induced colitis therapy. PMID- 16047472 TI - Intracranial tuberculomas in a nonimmuncompromised patient with abdominal tuberculosis misdiagnosed as Crohn's disease. PMID- 16047473 TI - Primary torsion of the greater omentum: a difficult diagnosis. PMID- 16047474 TI - Recurrent torsion of a giant Meckel's diverticulum. PMID- 16047475 TI - Nitric oxide-dependent osteopontin expression induces metastatic behavior in HepG2 cells. AB - Our objective was to delineate the role of nitric oxide (NO) in osteopontin (OPN) associated metastatic properties in HepG2 cells. OPN is the major phosphoprotein secreted by malignant cells in patients with advanced metastatic cancer, is frequently overexpressed in human tumors, and has been implicated as a key mediator of tumor cell metastasis. OPN is significantly overexpressed in hepatocellular cancer (HCC) and correlates with capsular infiltration and behavior. In addition, significantly increased inducible nitric oxide synthase (iNOS) and NO expression are found in HCC. In archived human samples of normal, cirrhotic, and HCC livers, we demonstrate that iNOS and OPN protein are strongly coexpressed in hepatoma cells. In the setting of cirrhosis, hepatocytes express iNOS, but not OPN. Further in vitro studies performed with HepG2 hepatocellular cancer cells demonstrate that exogenous NO transcriptionally upregulates OPN expression. Enhanced expression of OPN in this setting is associated with increased in vitro cell adhesion and invasion. These data suggest that NO enhances HCC expression of OPN and, as a result, conveys a metastatic phenotype. PMID- 16047476 TI - HTERT mRNA partially regulates telomerase activity in gastric adenocarcinoma and adjacent normal gastric mucosa. AB - The extent to which human telomerase reverse transcriptase (hTERT) mRNA and its splice variants control telomerase activity in human cancers is controversial. Telomerase and hTERT mRNA were assessed quantitatively in paired samples of gastric adenocarcinoma and adjacent normal tissue. Splice variants within the hTERT reverse transcriptase domain (alpha, beta, alphabeta) were detected by RT PCR. In gastric adenocarcinoma, compared to normal tissue, median telomerase activity increased significantly (from 0 total product generated [tpg; 95% confidence interval CI, 0-2.3] to 16.1 tpg [95% CI, 3.7-97]; P = 0.008) and median hTERT mRNA levels also increased (from 2.21 [95% CI, 1.40-4.62] to 7.08 [95% CI, 3.26-10.8]; P = 0.0054). hTERT mRNA and telomerase activity correlated in normal gastric mucosa (r = 0.819, P = 0.0002). Alpha, beta, and alphabeta deletions were similar in both groups. We conclude that hTERT mRNA partially regulates telomerase activity in normal gastric mucosa and gastric adenocarcinoma. In contrast, hTERT mRNA splicing is not involved in the regulation of enzyme activity. PMID- 16047477 TI - Effect of concomitant polyethylene glycol and celecoxib on colonic aberrant crypt foci and tumors in F344 rats. AB - We investigated whether celecoxib augments the protective effect of polyethylene glycol (PEG) on colonic aberrant crypt foci (ACF) and tumor formation in F344 rats treated with azoxymethane (AOM). Three groups of rats received AOM: I (AOM alone), II (PEG), and III (PEG/celecoxib). PEG reduced the mean number of total ACF per colon from 190 to 141 (P < 0.05; 26% reduction) and > or = 4-crypt ACF from 95 to 58 (P < 0.01; 39%). Group III rats had a greater proportion of their ACF distally; whereas transverse colon ACF were reduced approximately 50%, distal ACF were reduced by only approximately 8% (P < 0.05). Of 13 large bowel tumors, 8 were in Group I, 4 in Group II, and 1 in Group III rats (P = 0.02). Thus in AOM treated rats celecoxib appeared to enhance the PEG-induced reduction in colonic tumor formation, and in transverse but not distal or whole-colon ACF. PMID- 16047478 TI - Abdominal actinomycosis simulating malignancy of the right colon. PMID- 16047479 TI - Gastrointestinal stromal tumor (GIST) of the esophagus detected by positron emission tomography/computed tomography. PMID- 16047480 TI - Measurement of serum electrolytes and phosphate after sodium phosphate colonoscopy bowel preparation: an evaluation. AB - Our purpose was to observe the effects of sodium phosphate (NaP) colonoscopy preparation on serum electrolytes, phosphate, and calcium and to identify factors associated with any adverse effects. In an unselected group of 100 consecutive patients attending for out patient colonoscopy, 45% of patients had raised serum phosphate, which was positively correlated with creatinine and age. There was a negative association of phosphate with calcium; 16% of patients had hypocalcemia and 26% had hypokalemia. Patients taking ACE inhibitors, AT2 antagonists, or diuretics were associated with hyperphosphatemia. Significant electrolyte and metabolic disturbance from colonoscopy preparation has been shown with NaP preparation, without overt clinical effects. We recommend that elderly patients and those with significant comorbidity have their electrolytes and calcium measured, and diuretics and ACE inhibitors stopped, before NaP administration. Endoscopy units should be alert for patients who might be suffering from electrolyte disturbance postpreparation and be prepared to measure their electrolytes. PMID- 16047481 TI - Hazards of endoscopic biopsy for flat adenoma before endoscopic mucosal resection. PMID- 16047482 TI - Chronic gastric electrical stimulation for gastroparesis reduces the use of prokinetic and/or antiemetic medications and the need for hospitalizations. AB - To investigate the effect of chronic gastric electrical stimulation (GES) on the daily use of prokinetics and antiemetics, hospitalizations, total symptom score (TSS), SF-36 status for health-related quality of life (HQOL), and gastric emptying of a solid meal, we evaluated 37 gastroparetic patients preoperatively and 1 year after undergoing GES implant. Prokinetic and antiemetic use was significantly reduced. Of 27 patients on at least one prokinetic at baseline, 8 were off at 1 year. Twenty-six patients requiring antiemetics before surgery decreased to 17. Mean TSS was significantly reduced and the reduction for patients off medications was significantly better than for patients still on medications. Overall SF-36 scores for HQOL were significantly improved, and patients off antiemetics had a significantly higher HQOL score than for patients on antiemetics at 1 year. Hospitalizations decreased from 50 +/- 10 days for the year prior to GES therapy to 14 +/- 3 days (P < 0.05). However, gastric emptying was not significantly improved. Conclusions are as follows. (1) Chronic GES significantly reduced the use of prokinetic/antiemetic medications and the need for hospitalization in gastropraretic patients, whose clinical and quality of life outcomes also significantly improved. (2) These data provide evidence of the positive economic impact of this new therapy on long-term clinical outcomes in gastroparetic patients not responding to standard medical therapy. PMID- 16047483 TI - In vivo gastric and intestinal slow waves in W/WV mice. AB - The aim was to investigate whether there are regular gastric and intestinal slow waves in conscious W/WV mice. Eleven W/WV mice and 11 wild-type mice were implanted with two pairs of electrodes in the stomach and small intestine. Gastrointestinal slow waves were recorded both under anesthesia and in the conscious state. Atropine and verapamil were given separately to an additional 10 W/WV mice. Results were as follows. (1) The conscious W/WV mice showed lower rhythmic slow waves in the small intestine (77.1 vs 93.5%; P < 0.001). However, the frequency (10.7 vs 18.8 cpm; P < 0.0001) and the antregrade propagation of intestinal slow waves in W/Wv mice were significantly lower than in the controls. In the stomach, regular slow waves were recorded in both groups, with no difference between the two groups. (2) Anesthesia significantly impaired both gastric and intestinal slow waves in both groups. (3) Atropine and verapamil had no effects on the rhythmicity of intestinal slow waves. We conclude that ICC-MY may not be the sole pacemaker cells for slow waves in the small intestine. There may be some abnormality of smooth muscle cells in W/WV mice that causes a reduction in the frequency, rhythmicity, and antegrade propagation of slow waves. PMID- 16047484 TI - Gastrointestinal bleeding in a patient with Ehlers-Danlos syndrome: an endoscopic dilemma. PMID- 16047485 TI - Splenic vein aneurysm demonstrated by magnetic resonance angiography. PMID- 16047486 TI - Disruption of erythrocytes distinguishes fixed cells/tissues from viable cells/tissues following microwave coagulation therapy. AB - Microwave coagulation therapy (MCT) has recently been applied to treat hepatic tumors. However, the histological changes in the liver following MCT have not been fully elucidated. A type of cell death known as microwave fixation has been reported in areas adjacent to the microwave irradiator electrodes, and these areas are without acid phosphatase (AcP) activity. Diagnosis of microwave-fixed tissue by hematoxylin and eosin (HE) staining is very difficult because morphology is well maintained for months. In an effort to clarify the histological changes and the mechanisms of microwave fixation, we performed HE staining, enzyme histochemistry for AcP, and electron microscopy in both rat and human liver samples after MCT. Although the microwave-fixed tissues maintained their structure on HE staining, membranes of microwave-fixed cells were seriously damaged and there were no apparent organelle structures in these cells on electron microscopy. Erythrocytes were also damaged in these tissues on both light and electron microscopy. The cause of microwave fixation is thought to be injury of the membrane, which is similar to coagulative necrosis. In conclusion, microwave fixation can be considered a type of coagulative necrosis without enzyme digestion. Disruption of erythrocytes on HE staining is an interesting and important diagnostic clue in distinguishing nonviable fixed tissues from viable tissues following MCT. PMID- 16047487 TI - Chronic albumin infusions to achieve diuresis in patients with ascites who are not candidates for transjugular intrahepatic portosystemic shunt (TIPS). AB - While transjugular intrahepatic portosystemic shunt (TIPS) is a common therapy for cirrhotic patients with diuretic-resistant or diuretic-refractory ascites, some patients are unsuitable for the procedure for technical or medical reasons. We report our experience with the use of chronic intravenous albumin infusions to achieve diuresis in this difficult patient population and review the historic experience of chronic albumin infusions as a treatment for ascites. Nineteen patients with cirrhosis and diuretic-resistant or diuretic-refractory ascites who were deemed unsuitable for TIPS received outpatient intravenous albumin infusions (50 g) weekly for at least 4 weeks. The following endpoints were retrospectively recorded: serum sodium, serum creatinine, blood urea nitrogen, hematocrit, bilirubin, albumin, international normalized ratio, body weight, and Model for End-stage Liver Disease (MELD) score. The contraindications for TIPS included the following: portal vein thrombosis, two; advanced age, one; encephalopathy, nine; hyperbilirubinemia, five; and other, two. Compared to pretreatment, posttreatment weight decreased in 17 patients, remained unchanged in 0 patients, and increased in 2 patients. The overall mean change in body weight (before vs. after therapy) was 8 lb (P < 0.05). The only significant change in biochemistry was an increase in serum albumin from 2.5 g/dl before therapy to 3.5 g/dl after therapy (P < 0.05). We conclude that (1) recurrent intravenous weekly albumin infusions resulted in significant loss of edema and ascites as measured by loss of body weight, and (2) clinicians may want to consider chronic albumin infusions for selected patients with refractory ascites who are not candidates for TIPS. PMID- 16047488 TI - Hepatitis C virus core protein modulates fatty acid metabolism and thereby causes lipid accumulation in the liver. AB - We studied the roles of hepatitis C virus (HCV) core protein in hepatic steatosis and changes in hepatic lipid metabolism. HCV core protein expression plasmid was transfected in HepG2. Triacylglyceride (TG) and mRNA level associated with lipid metabolism were measured. Male C57BL/6 mice were infected with HCV core recombinant adenovirus and used for lipids and mRNA studies. In HCV core protein expressing cells, peroxisome proliferator-activated receptor (PPAR)alpha, multidrug resistance protein (MDR) 3, and microsomal triglyceride transfer protein (MTP) were down-regulated 48 hr after transfection. In HCV core protein expressing mice, hepatic TG content and hepatic thiobarbituric acid-reactive substances increased. PPARalpha, MDR2, acyl-CoA oxidase (AOX), and carnitine palmitoyl transferase-1 (CPT-1) were down-regulated. HCV core protein down regulated lipid metabolism-associated gene expression, Mdr2, CPT, and AOX, accompanied by down-regulation of PPARalpha. There findings may contribute to the understanding of HCV-related steatosis, induction of reactive oxygen species, and carcinogenesis. PMID- 16047489 TI - Cystinosis as a cause of noncirrhotic portal hypertension. AB - Cystinosis is a rare autosomal recessive storage disorder, characterized by the abnormal accumulation of cystine in cellular lysosomes. This accumulation, which can occur in any organ system, leads to crystallization of trapped cystine and ultimately cellular death. Hepatic manifestations of Cystinosis although rare, have been described in the literature. However, to our knowledge, only one other case of non-cirrhotic portal hypertension secondary to cystine accumulation in Kupffer cells has been reported. In this case and ours, portal hypertension was found in the absence of bridging fibrosis. Furthermore, in our case, for the majority of the patient's course, hepatic synthetic function remained normal. Cysteamine is therapeutic in this disorder, and can lead to significant removal of cystine, and thus to reversibility of disease, however, it requires high doses and must be taken regularly. Porto-systemic shunting in combination with aggressive medical therapy could potentially benefit patients who develop non cirrhotic portal hypertension in this clinical setting. PMID- 16047490 TI - Genetic polymorphisms of GSTT1, GSTM1, GSTP1, MnSOD, and catalase in nonhereditary chronic pancreatitis: evidence of xenobiotic stress and impaired antioxidant capacity. AB - Epidemiological studies have demonstrated a variety of potential environmental factors that may alter susceptibility to chronic pancreatitis (CP) through oxidative/xenobiotic stress; however, a direct causal and mechanistic role has not been established. We aimed (1) to determine the prevalence of functional genetic polymorphisms in the antioxidant enzymes, glutathione S-transferase GSTM 1, GSTP-1, and GSTT-1, manganese superoxide dismutase, and catalase in CP and (2) to reveal evidence of oxidative stress in patients with CP by measuring whole blood glutathione redox status. In total, 122 patients with CP (75 alcohol induced [A1CP], 33 idiopathic [ICP], and 13 hereditary) and 245 age- and sex matched controls were recruited. The prevalence of the functional GSTT-1 genotype (GSTT-1*A) was significantly higher in CP (88.5%) compared to healthy controls (76%; chi2 = 7.26, P = 0.007). Stratification to disease etiology demonstrated that the GSTT-1*A genotype was also significantly more prevalent among patients with ICP (94%; P = 0.02; 95% CI, 0.04-9.16) but not in those with A1CP. In 22 patients with stable CP, the whole-blood glutathione concentration (median [IQR]: 72 micromol/L [21-181 micromol/L]) and the glutathione redox ratio (GSH/GSSG) (median [IQR]: 9 (3-77]) were significantly reduced compared to those in 20 healthy volunteers (median [IQR]: 815 micromol/L [679-1148 micromol/L], P < 0.001, and 96 [52-347], P = 0.005, respectively). We conclude that the GSTT-1 functional genotype is associated with ICP. Evidence of altered glutathione redox status suggests that this disease modification may be a consequence of oxidative stress or the bioactivation of xenobiotics. PMID- 16047491 TI - Giant intrahepatic biliary cystadenoma in a male: a case report, immunohistopathological analysis, and review of the literature. PMID- 16047492 TI - Implementation of quality control methods in conjunction with chemometrics toward authentication of dairy products. AB - The implementation of novel and accurate quality and safety control methods in conjunction with chemometrics in various fields of science, particularly in food science, showed that this combination stands for a very powerful tool for detecting food authenticity. The latter reflects both geographic origin and variety. Dairy products, in particular, have repeatedly worried the public authorities in terms of authentication regarding origin and in view of the many illnesses occasionally due to products of specific origin. Therefore, the development of a robust and reliable system endowed with this discriminatory power (varietal and/or geographic) is of great importance, both in terms of public health and consumer protection. PMID- 16047493 TI - Colon targeted delivery systems: review of polysaccharides for encapsulation and delivery. AB - Colon-targeted delivery of bioactives has recently gained importance in addressing specific needs in the therapy of colon-based diseases. Many approaches have been attempted for the development of colon-specific delivery systems, with not much success in the past. Recent research into the utilization of the metabolic activity and the colonic microenvironment in the lower gastrointestinal tract has attained great value in the design of novel colon-targeted delivery systems based on natural biodegradable polymers. In the current article, special emphasis has been placed on polysaccharide systems, with minimal chemical modification, that have been exploitedfor colon targeting. These polysaccharide based encapsulation and targeted delivery systems are envisaged to have an immense potential for the development of food/nutraceutical formulations for colon-based diseases, including colorectal cancer. PMID- 16047494 TI - The role of nutrition in chronic renal insufficiency of childhood: how much do we know? AB - Dietary protein restriction in the treatment of symptomatic renal failure has been utilized for many years, especially as a means for reduction of 'fixed acid" load. Studies in animal models of renal failure suggest that low protein intake may retard the progression of renal disease as well. However, large, well organized investigations into this question in humans have fallen prey to difficulties that are almost impossible to overcome. Chief among these difficulties is the problem of chronically reducing protein intake in patients with a lifelong intake far above the recommended daily allowance (RDA). Another is the fact that all previous studies have been performed in patients with moderate to severe compromise of renal function. Thus, the potential efficacy of reduced protein intake in the retardation of the progression of renal disease remains an open question. In this article, we discuss the current state of knowledge and propose an approach to answering this question. PMID- 16047495 TI - Winemaking biochemistry and microbiology: current knowledge and future trends. AB - The fermentation of grape must and the production of premium quality wines are a complex biochemical process that involves the interactions of enzymes from many different microbial species, but mainly yeasts and lactic acid bacteria. Yeasts are predominant in wine and carry out the alcoholic fermentation, while lactic acid bacteria are responsible for malolactic fermentation. Moreover, several optional winemaking techniques involve the use of technical enzyme preparations. Considerable progress has been made recently in understanding the biochemistry and interactions of enzymes during the winemaking process. In this study, some of these recent contributions in the biochemistry of winemaking are reviewed. This article intends to provide an updated overview (including works published until December, 2003) on the main biochemical and microbiological contributions of the different techniques that can be used in winemaking. As well as considering the transformations that take place in traditional winemaking, the production of special wines, such as sparkling wines, 'sur lie' wines, and biologically aged wines, are also studied. PMID- 16047496 TI - Dietary polyphenols and the prevention of diseases. AB - Polyphenols are the most abundant antioxidants in the diet and are widespread constituents of fruits, vegetables, cereals, dry legumes, chocolate, and beverages, such as tea, coffee, or wine. Experimental studies on animals or cultured human cell lines support a role of polyphenols in the prevention of cardiovascular diseases, cancers, neurodegenerative diseases, diabetes, or osteoporosis. However, it is very difficult to predict from these results the effects of polyphenol intake on disease prevention in humans. One of the reasons is that these studies have often been conducted at doses or concentrations far beyond those documented in humans. The few clinical studies on biomarkers of oxidative stress, cardiovascular disease risk factors, and tumor or bone resorption biomarkers have often led to contradictory results. Epidemiological studies have repeatedly shown an inverse association between the risk of myocardial infarction and the consumption of tea and wine or the intake level of some particular flavonoids, but no clear associations have been found between cancer risk and polyphenol consumption. More human studies are needed to provide clear evidence of their health protective effects and to better evaluate the risks possibly resulting from too high a polyphenol consumption. PMID- 16047497 TI - Electrodialytic phenomena and their applications in the dairy industry: a review. AB - Electrodialysis (ED) is an electrochemical separation process by which electrically-charged species are transportedfrom one solution to another ED is a combined method of dialysis and electrolysis and can be performed with two main cell types: multi-membrane cells for dilution-concentration and water dissociation applications (membrane phenomena), and electrolysis cells for redox reactions (electrode phenomena). The dilution-concentration principle applications in the dairy industry consist mainly of the demineralization of milk or milk by-products. The use of ED with monopolar membrane for protein separation and acid caseinate production, and in bioreactors for organic acid production, is also studied in the dairy industry. The interest of ED as a membrane process has been triggered recently by the development of a new membrane type, bipolar membrane. This membrane carries out the dissociation of water molecules. ED with bipolar membranes was applied very recently to the production of lactic acid from whey product fermentation, production of caseinates, and fractionation of whey proteins. Two principle applications of electrode reactions were published: electrochemical coagulation (EC) to precipitate milk proteins, and electroreduction for the reduction of disulfide bonds in the proteins. It appears in this article that processes using membrane phenomena are more numerous and developed than electrolytic applications. This is the composition of milk and the lack of knowledge of redox reactions of the different food compounds that limit the applications and the development of electrolytic phenomena. Electrodialytic phenomena present a great potential for application in the dairy industry, and more generally, in the food industry; many of these applications have to be discovered. PMID- 16047498 TI - Are factor analytical techniques used appropriately in the validation of health status questionnaires? A systematic review on the quality of factor analysis of the SF-36. AB - Factor analysis is widely used to evaluate whether questionnaire items can be grouped into clusters representing different dimensions of the construct under study. This review focuses on the appropriate use of factor analysis. The Medical Outcomes Study Short Form-36 (SF-36) is used as an example. Articles were systematically searched and assessed according to a number of criteria for appropriate use and reporting. Twenty-eight studies were identified: exploratory factor analysis was performed in 22 studies, confirmatory factor analysis was performed in five studies and in one study both were performed. Substantial shortcomings were found in the reporting and justification of the methods applied. In 15 of the 23 studies in which exploratory factor analysis was performed, confirmatory factor analysis would have been more appropriate. Cross validation was rarely performed. Presentation of the results and conclusions was often incomplete. Some of our results are specific for the SF-36, but the finding that both the application and the reporting of factor analysis leaves much room for improvement probably applies to other health status questionnaires as well. Optimal reporting and justification of methods is crucial for correct interpretation of the results and verification of the conclusions. Our list of criteria may be useful for journal editors, reviewers and researchers who have to assess publications in which factor analysis is applied. PMID- 16047499 TI - Content comparison of health-related quality of life (HRQOL) instruments based on the international classification of functioning, disability and health (ICF). AB - The increasing recognition of the patient perspective and, more specifically, functioning and health, has led to an impressive effort in research to develop concepts and instruments to measure them. Health-Related Quality of Life (HRQOL) and the International Classification of Functioning Disability and Health (ICF) represent two different perspectives from which to look at functioning and health. Therefore, it is expected that both will often be used concurrently in clinical practice, research and health reporting. The objective of our study was to examine the relationship between six HRQOL instruments (the SF-36, the NHP, the QL-I, the WHOQOL-BREF, the WHODASII and the EQ-5D) and the ICF. All six HRQOL instruments were linked to the ICF separately by two trained health professionals according to ten linking rules developed specifically for this purpose. The degree of agreement between health professionals was calculated by means of the kappa statistic. Bootstrapped confidence intervals were calculated. In the 148 items of the 6 instruments a total of 226 concepts were identified and linked to the ICF. The estimated kappa coefficients range between 0.82 and 0.98. The concepts contained in the items of the HRQOL instruments were linked to 91 different ICF categories, 17 categories of the component body functions, 60 categories of the component activities and participation, and 14 categories of the component environmental factors. Twelve concepts could not be linked to the ICF at all. In the component body functions, only emotional functions are covered by all examined instruments. In the component activities and participation, all instruments cover aspects of work, but the half of them scarcely cover aspects of mobility. Only four of the six instruments address environmental factors. The ICF proved highly useful for the comparison of HRQOL instruments. The comparison of selected HRQOL instruments may provide clinicians and researchers with new insights when selecting health-status measures for clinical studies. PMID- 16047500 TI - Comparison of health-related quality of life questionnaires in ambulatory oncology. AB - The purpose of this study is to compare three commonly used health-related quality of life (HR-QOL) questionnaires for their ease of use, accuracy, and patient preference; identify factors related to patient preference; identify differences in patient completion rates; and to identify factors associated with patient completion of these questionnaires. Three psychometrically sound measures, the Symptom Distress Scale (SDS), Medical Outcome Study Short Form-36 (SF-36), and Functional Assessment of Cancer Therapy (FACT), were tested. Seventy nine patients completed questionnaires in the ambulatory oncology setting. No significant differences in patient ratings were found in ease of use and accuracy among the questionnaires. All of the questionnaires were rated as easy to use and accurate. Patient ratings on preference were marginally significant (p = 0.07). Forty-six percent of participants indicated that they preferred the SDS, whereas 27% and 39% preferred the SF-36 and the FACT. No significant differences in patient completion rates were found among the questionnaires. One hundred percent completion rates ranged from 88.6% for the SDS to 78.5% for the SF-36, and 80% completion rates ranged from 98.7% for the SDS to 94.9% for the SF-36. Administration of standardized HR-QOL questionnaires is feasible in the clinical setting. PMID- 16047501 TI - Comparing SF-36 scores across three groups of women with different health profiles. AB - BACKGROUND: The widespread use of the Medical Outcomes Study (MOS) 36-item Short Form Health Survey (SF-36) facilitates the comparison of health-related quality of life (HRQL) across independent studies. OBJECTIVES: To compare the scores of eight scales and two summary scales of the SF-36 across participants in the Women's Healthy Eating and Living (WHEL) trial, the Women's Health Initiative Dietary Modification trial (WHI-DM), and the MOS, and to illustrate the use of effect sizes for interpreting the importance of group differences. METHODS: WHEL and WHI-DM are both multi-center dietary interventions; only data from the UC Davis sites were used in our study. WHEL participants had a recent history of breast cancer, WHI-DM participants were healthy, postmenopausal women, and women in the MOS had a history of hypertension, diabetes, heart disease, or depression. General linear models were used to identify statistically significant differences in scale scores. Meaningful differences were determined by effect sizes computed using a common within-group standard deviation (SD) and SDs from normative data. RESULTS: After adjusting for age and marital status, SF-36 scores for the WHI-DM and WHEL samples were similar and both had statistically significantly higher scores than the MOS sample. Relative to the WHEL or WHI-DM studies, MOS scores for scales related to the physical domain were clearly meaningfully lower whereas scale scores related to the mental health domain were potentially meaningfully lower. CONCLUSIONS: The HRQL of breast cancer survivors is comparable to that of healthy women and better than that of women with chronic health conditions, particularly with respect to physical health. This study illustrated the use of ranges of effects sizes for aiding the interpretation of SF-36 scores differences across independent studies. PMID- 16047502 TI - Performance of the SF-36 health survey in screening for depressive and anxiety disorders in an elderly female Swedish population. AB - OBJECTIVE: To assess the sensitivity, specificity and predictive validity of suggested cut-off scores in the SF-36 mental health (MH) and mental component summary (MCS) in screening for depressive and anxiety disorders in a population sample of older Swedish women. METHOD: The sample comprised 586 randomly selected females aged 70-84 years who took part in an in-depth psychiatric examination. This provided the 'gold standard' against which the usefulness of SF-36 recommended thresholds for screening for depressive and anxiety disorders in older Swedish women was examined. RESULTS: Based on DSM-III-R criteria, 69 women (12%) were diagnosed with depression (major depression, dysthymia and/or depression NOS) and 49 (8%) with generalised anxiety and panic disorders. The previously recommended MH and MCS cut-offs (i.e. 52 and 42) gave a specificity for diagnosis of depression of 92 and 82% and sensitivity of 58 and 71%, respectively. Both the MH and MCS were good predictors of depressive disorders but poor predictors of anxiety disorders. CONCLUSION: The study supports the predictive validity of suggested SF-36 MH and MCS cut-off scores in screening for depressive disorder but not for anxiety disorder in older women in Sweden. PMID- 16047503 TI - The effects of antipsychotic treatment on quality of life of schizophrenic patients under naturalistic treatment conditions: an application of random effect regression models and propensity scores in an observational prospective trial. AB - The study examines the effect of different types of antipsychotic treatment on the health related quality of life (HRQL) of people with schizophrenia under naturalistic outpatient treatment conditions. In a prospective study design, 307 schizophrenic patients were followed over a period of 2.5 years. HRQL, clinical characteristics, and type of antipsychotic medication were assessed five times every 6 months. HRQL was assessed by the SF-36. Random effect regression models were computed for the SF-36 mental (MCS) and physical (PCS) component scores. Propensity scores were included in the regression models to reduce a possible sample selection bias. Monotherapeutic treatment with new atypical neuroleptic drugs had a more positive effect on the mental health related quality of life (MCS) in comparison to treatment with polypharmacological treatment but not with oral conventional antipsychotics. Monopharmaceutical treatment with depot antipsychotic drugs had a more positive effect on the physical health related quality of life (PCS) in comparison to polypharmacological treatment. Study results indicate that atypical antipsychotic drugs are not superior to conventional antipsychotics with regard to the effect on QOL. However, monopharmaceutical treatment can be assumed to be more effective in improving mental and physical related QOL than polypharmaceutical treatment. PMID- 16047504 TI - Assessment of quality of life in later life: development and validation of the QuiLL. AB - The aim was to develop a quality of life (QOL) instrument, informed by older people, carers and professionals in older peoples' services, for use by community care staff as part of their assessment, care-planning and outcome monitoring procedures. The multi-phase development project involved: qualitative interviews to generate the item pool; pre-testing; preliminary field-testing; and final testing in a community survey and in health and social care settings. The process was informed by over 100 interviews with older people, carers, professionals, academics and policy-makers. Two products emerged following data-reduction: a research instrument (64 items), and a shorter assessment tool suitable for routine use in clinical, therapeutic or case-management practice (27 items, taking 7-15 min to complete). A community survey using the research instrument achieved a 71% response rate (n = 249). Ninety six percent of people found the domain content of the assessment tool relevant, and 80% considered the items covered were important. Both instruments have good internal consistency (alpha = 0.85). Inter-rater reliability was good for research staff, but poor between them and operational staff. Little objective change took place during the 3-month follow-up study, but where it did the direction was consistent with subjective change. The instrument needs to be applied in different contexts to assess interventions of known impact. Together with daily living and health status measures it can form part of a comprehensive assessment for older people. PMID- 16047505 TI - Health-related quality of life of patients with HIV: impact of sociodemographic, clinical and psychosocial factors. AB - OBJECTIVES: This study aims to analyse how a wide group of clinical, social, demographic and psychological factors are related to both physical and mental quality of life in HIV + patients. DESIGN: A cross-sectional study was carried out of 320 HIV + patients in antiretroviral treatment who attended infectious diseases units in four hospitals in the region of Andalusia (Spain). METHODS: Health-Related Quality of Life was measured by the MOS-HIV. Included as independent variables were: sociodemographic variables, variables related to antiretroviral therapy, psychosocial variables like social support (Duke-UNC-11) and psychological morbidity (GHQ-28), variables related to main risk behaviours and clinical variables. RESULTS: In the multiple linear regression analysis, a better PHS quality of life was found to be associated with the absence of mental illness, social support, not being an intravenous drug user and using more than one type of non-injectable drug. A better quality of life, in mental terms, was found to be associated with fewer years as a non-intravenous drug user, having social support, absence of mental illness, not being an intravenous drug user taking only one additional pill, not having any difficulty in taking the medication, and being female. CONCLUSIONS: The study of other non-biological factors that may be related to quality of life has been limited practically to social support and the emotional state. This study highlights the importance of these factors independently from the clinical state, as well as the existence of other psychological and behavioural factors that are also related. PMID- 16047506 TI - Health-related quality of life deficits associated with diabetes and comorbidities in a Canadian National Population Health Survey. AB - OBJECTIVE: To assess the impact of comorbid heart disease, stroke and arthritis on health-related quality of life (HRQL) in people with diabetes in the general Canadian population. METHODS: Data were collected as part of the 1996-1997 Canadian National Population Health Survey. HRQL was assessed using overall Health Utilities Index Mark 3 (HUI3) and single attribute utility scores. Respondents (N = 66,093) were classified into 1 of 16 groups based on the presence or absence of diabetes, heart disease, stroke, and arthritis, in all possible combinations and HRQL scores were compared using analysis of covariance. RESULTS: Overall HUI3 scores for respondents with diabetes alone (0.88, 95% CI: 0.87-0.89) were lower than controls (0.92: 95% CI: 0.92-0.92, p < 0.001). Overall HUI3 scores for diabetes in combination with heart disease (0.77, 95% CI: 0.74 0.79), arthritis (0.78, 95% CI: 0.77-0.79) or stroke (0.79, 95% CI: 0.74-0.85) were considerably lower than diabetes alone. Triplets of comorbidities were associated with overall HRQL deficits of approximately 0.26-0.30, relative to controls. CONCLUSIONS: The illness burden experienced by individuals with diabetes is not only associated with diabetes itself, but largely with comorbid medical conditions. PMID- 16047507 TI - Comparison of EQ-5D, HUI, and SF-36-derived societal health state values among spine patient outcomes research trial (SPORT) participants. AB - PURPOSE: To compare societal values across health-state classification systems and to describe the performance of these systems at baseline in a large population of persons with confirmed diagnosis of intervertebral disc herniation (IDH), spinal stenosis (SpS), or degenerative spondylolisthesis (DS). METHODS: We compared values for EQ-5D (York weights), HUI (Mark 2 and 3), SF-6D, and the SF 36-derived estimate of the Quality of Well Being (eQWB) score using signed rank tests. We tested each instrument's ability to discriminate between health categories and level of symptom satisfaction. Correlations were assessed with Spearman rank correlations. We evaluated ceiling and floor effects by comparing the proportion at the highest and the lowest possible score for each tool. In addition, we compared proportions at the highest and lowest levels by dimension. The number of unique health states assigned was compared across instruments. We calculated the difference between those who were very dissatisfied and all others. RESULTS: Mean values ranged from 0.39 to 0.63 among 2097 participants ages 18-93 (mean age 53, 47% female) with significant differences in pair-wise comparisons noted for all systems. Correlations ranged from 0.30 to 0.78. Although all systems showed statistically significant differences in health state values when baseline comparisons were made between those who were very dissatisfied with their symptoms and those who were not, the magnitude of this difference ranged widely across systems. Mean differences (95% CI) between those very dissatisfied and all others were 0.30 (0.269, 0.329) for EQ-5D, 0.22 (0.190, 0.241) for HUI(3), 0.18 (0.161, 0.201) for HUI(2), 0.11 (0.095, 0.117) for SF-6D, 0.04 (0.039, 0.049) for eQWB, and 0.07 (0.056, 0.077) for VAS (with transformation applied to group means). CONCLUSION: Differences in preference weighted health state classification systems are evident at baseline in a population with confirmed IDH, SpS, and DS. Caution should be used when comparing health state values derived from various systems. PMID- 16047508 TI - Are indirect utility measures reliable and responsive in rheumatoid arthritis patients? AB - BACKGROUND: Preference-based, generic measures are increasingly being used to measure quality of life and as sources for quality weights in the estimation of Quality Adjusted Life Years (QALYs) in rheumatoid arthritis (RA). However, among the most commonly used instruments (the Health Utilities Index 2 and 3 [HUI2 and HUI3], the EuroQoL-5D [EQ-5D], and the Short Form-6D [SF-6D], there has been little comparative research. Therefore, we examined the reliability and responsiveness of these measures and the Rheumatoid Arthritis Quality of Life (RAQoL) and the Health Assessment Questionnaire (HAQ) in a sample of RA patients. MAJOR FINDINGS: Test-retest reliability was acceptable for all of the instruments with the exception of the EQ-5D. Using two external criteria to define change (a patient transition question and categories of the patient global assessment of disease activity VAS), the RAQoL was the most responsive of the instruments. For the indirect utility instruments, the HUI3 and the SF-6D were the most responsive for measuring positive change. On average, for patients whose RA improved, the absolute change was highest for the HUI3. CONCLUSIONS: The HUI3 and the SF-6D appear to be the most responsive of the preference-based instruments in RA. However, differences in the magnitude of the absolute change scores have important implications for cost-effectiveness analyses. PMID- 16047509 TI - Quality of life of children with language delays. AB - We investigated health-related quality of life (HRQOL) of children with language problems and controls. Data on language development (Language Screening Instrument 3-years-olds, Van Wiechen items) and HRQOL by means of the TNO-AZL Pre school children Quality of Life-questionnaire (TAPQOL) were collected at age 3 in a population-based cohort by parental questionnaire (n = 8877, response 78%; mean age 39.1 months (SD 2.0), 4347 were girls). Cronbach's alpha (internal consistency) ranged between 0.63 and 0.85. Dependent on the definition of language problem, 131 to 316 children appeared to be language impaired. Receiver Operating Characteristic analyses (ROC-curves) to assess the discriminative ability of six TAPQOL scales revealed that the Communication scale and Social Functioning scale discriminated best between children with language problems and children without these problems. Language-impaired children had significantly lower scores on the Communication scale and Social Functioning scale as compared to children without language problems (p < 0.01). The findings indicate that language problems at age three can have an impact on children's social life. These results provide additional evidence for the importance of monitoring the language development and its consequences during childhood. PMID- 16047510 TI - The situational fatigue scale: a different approach to measuring fatigue. AB - Fatigue can be defined as a subjective state of an imbalance in the availability of inner resources needed to perform physical or mental activities. The level of fatigue is determined not only by the availability of inner resources but also by the demands of the activities performed. Most conventional fatigue scales require subjects to rate their level of fatigue without specifying the situation. In the present study, we constructed a subjective rating scale, the Situational Fatigue Scale (SFS), with which subjects estimated their level of fatigue in specific activities of daily life. We administered the SFS, along with the Fatigue Assessment Instrument (FAI) to 96 outpatients in a family-medicine clinic and to 62 college students to assess the psychometric properties of the SFS. Principle component analysis revealed two underlying factors: physical fatigue and mental fatigue. SFS scores were significantly correlated with several FAI scores and differentiated patients complaining of fatigue from those who did not. The SFS also showed good internal consistency and test-retest reliability. These results suggest that the SFS could be a useful tool to measure a different dimension of the broad concept of fatigue. PMID- 16047511 TI - An assessment of the feasibility and utility of the MS symptom and impact diary (MSSID). AB - The Multiple Sclerosis Symptom and Impact Diary (MSSID) was developed to provide a more comprehensive assessment of the impact of multiple sclerosis (MS) on HRQoL for use in clinical practice. This paper reports on an assessment of its feasibility and utility in two outpatient samples of people with MS (n = 13 and n = 63) using quantitative and qualitative methods. The response rate in study 2 was 82% and 83% of days were fully completed. Most respondents found the MSSID easy to understand and got into the habit of completing it. Missing items increased over time and those who experienced a relapse had more missing items than those who did not but there was no difference in the number of missed days. Some respondents found completing the MSSID enabled them to manage their lives more effectively and provided useful information to their neurologist. It is concluded that the MSSID is feasible for people with MS to complete and some may find the MSSID helpful as a tool to monitor their condition. Further research is needed to examine clinicians' perceptions of the feasibility and utility of the MSSID within clinical practice. PMID- 16047512 TI - Development and validation of a new health-related quality of life instrument for patients with sinusitis. AB - BACKGROUND AND OBJECTIVE: There are few validated measures of sinusitis-specific health-related quality of life (HRQL). This study used patient focus and pretesting groups followed by a prospective cohort study to develop and validate a HRQL instrument for patients with sinusitis. METHODS: Instrument development involved a systematic literature review, use of expert input, and patient focus and pretesting groups. Patients were recruited from the practices of primary care providers and otolaryngologists. The derived survey instrument then underwent prospective testing in patients with acute sinusitis, chronic sinusitis, allergic rhinitis, and asymptomatic controls. Reduced item scales of the original instrument were developed for symptom frequency and bothersomeness. The psychometric properties of the survey instrument were evaluated for reliability, construct validity, responsiveness, and interpretability. RESULTS: In the prospective study, 47 patients with acute sinusitis and 50 patients with chronic sinusitis were compared to 18 patients with allergic rhinitis and 60 patients without nasal symptoms. Forty-three (91.5%) patients with acute sinusitis completed the questionnaire at baseline and at 1-month follow-up. Internal consistency was high for the symptom impact scale for acute and chronic sinusitis patients. The symptom frequency and especially bothersomeness scales had lower internal consistency particularly for acute sinusitis patients. Reproducibility among surgical patients retested prior to their procedure was good for each scale. A high degree of disciminant validity was demonstrated when comparing sinusitis patients to other groups, and a high degree of convergent validity was seen when the new measures were compared to other HRQL measures at baseline. Among patients with acute sinusitis, the responsiveness and interpretability of the symptom frequency, bothersomeness and impact scales were excellent. CONCLUSIONS: This study developed and validated a new sinusitis-specific HRQL instrument. The instrument included symptom frequency, bothersomeness and impact scales. It was shown to be valid in patients with acute and chronic sinusitis, and highly responsive and interpretable in acute sinusitis patients managed in the primary care setting. PMID- 16047513 TI - Development and psychometric validation of a new patient satisfaction instrument: the osteoARthritis Treatment Satisfaction (ARTS) questionnaire. AB - Given the increasing interest in treatment satisfaction research and the lack of a specific questionnaire in osteoarthritis (OA), we developed and explored the psychometric properties of the osteoARthritis Treatment Satisfaction (ARTS) questionnaire. The ARTS questionnaire which consists of 18 items was developed in French following the analysis of semi-structured interviews performed among 20 OA participants, five rheumatologists and five general practitioners. Psychometric properties were assessed in France on a cross-sectional sample of 797 OA participants and test-retest reliability was evaluated in an independent sample of 111 clinically stable OA participants who filled-in the questionnaire within a 7.7 (+/- 3.1) day interval. Using principal component analysis, four scales were identified: Treatment advantages (seven items), Treatment convenience (three items), Treatment confidence (two items) and Satisfaction with physician (six items). Item convergent and item discriminant validity were satisfactory. Internal consistency provided evidence of reliability and lack of redundancy (Cronbach's alphas ranged from 0.66 to 0.86). Test-retest reliability was acceptable for two out of four scales (intraclass correlations coefficients (ICC) ranged from 0.61 to 0.75). Significant between groups differences were found on the ARTS scales, demonstrating the known groups validity of the ARTS questionnaire. The responsiveness of the ARTS is still to be documented. PMID- 16047514 TI - Cross-cultural adaptation and validation of the Korean version of the EQ-5D in patients with rheumatic diseases. AB - OBJECTIVES: This study aimed to determine the cross-cultural adaptation and validation of the Korean version of the EQ-5D in rheumatic conditions. METHODS: Translation, back-translation and cognitive debriefing were performed according to the EuroQol group's guidelines. For validity, 508 patients were recruited and administered the EQ-5D, Short-Form 36 and condition-specific measures. Construct validity and sensitivity were evaluated by testing a-priori hypotheses. For reliability, another 57 patients repeated the EQ-5D at 1-week interval, and intra class correlations (ICC) and kappa statistics were estimated. For responsiveness, another 60 patients repeated it at 12-week interval within the context of clinical trial, and standardized response mean(SRM) were calculated. RESULTS: The cross-cultural adaptation produced no major modifications in the scale. The associations of the EQ-5D with the generic- and condition-specific measures were observed as expected in hypotheses: the higher EQ-5Dindex and EQ-5D(VAS) scores, the better health status by generic- or condition-specific measures, and the better functional class. The ICCs were 0.751 and 0.767, respectively, and kappa ranged from 0.455 to 0.772. The SRM were 0.649 and 0.410, respectively. CONCLUSION: The Korean EQ-5D exhibits good validity and sensitivity in various rheumatic conditions. Although its reliability and responsiveness were not excellent, it seems acceptable if condition-specific measures are applied together. PMID- 16047515 TI - Translation and cultural adaptation of Health Utilities Index (HUI) Mark 2 (HUI2) and Mark 3 (HUI3) with application to survivors of childhood cancer in Brazil. AB - INTRODUCTION: There are few publications reporting health-related quality of life (HRQL) in developing nations. Most instruments measuring HRQL have been developed in English-speaking countries. These instruments need to be culturally adapted for use in non-English-speaking countries. The HUI2 and HUI3 are generic, preference-based systems for describing health status and HRQL. Developed in Canada, the systems have been translated into more than a dozen languages and used worldwide in hundreds of studies of clinical and general populations. METHODS: The Brazilian-Portuguese translation of the HUI systems was supervised by senior HUInc staff having experience with both the HUI systems and translations. The process included two independent forward translations of the multi-attribute health status classification systems and related questionnaires, consensus between translators on a forward translation, back-translation by two independent translators of the forward translation, and review of the back translations by original developers of the HUI. The final questionnaires were tested by surveying a sample of convenience of 50 patients recruited at the Centro de Tratamento e Pesquisa-Hospital do Cancer in Sao Paulo, Brazil. RESULTS: Fifty patients were enrolled in the study. No assessor, patient or nurse or physician, reported problems answering the HUI questionnaires. No significant differences were found in mean overall HUI2 or HUI3 utility scores among types of assessors. Variability in scores are similar to those from other studies in Latin America and Canada. CONCLUSION: Test results provide preliminary evidence that the Brazilian-Portuguese translation is acceptable, understandable, reliable and valid for assessing health-status and HRQL among survivors of cancer in childhood in Brazil. PMID- 16047516 TI - Development and validation of the interview version of the Hong Kong Chinese WHOQOL-BREF. AB - INTRODUCTION: The Hong Kong Chinese version of the WHOQOL-BREF was designed as a self-administered questionnaire and has limitations in clinical application on subjects who have limitations in reading or writing. An interview version is therefore needed to avoid sampling biases in clinical studies. Since there are significant differences in the written Chinese and spoken Cantonese, which is a dialect commonly spoken among people in Hong Kong, and adaptation process for converting the written Chinese into spoken Cantonese was necessary. The interview version was designed to allow administration in both face-to-face interview and telephone interview mode. METHODS: Three members of the research team translated the formal written Chinese in the self-administered version of the WHOQOL BREF(HK) into colloquial Cantonese separately. Brief notes extracted from the facet definitions of the WHOQOL-100 were added in brackets after some questions to further explain the intention of the questions. Two series of focus groups were conducted and subsequently the field test version was produced. 329 subjects were recruited by convenient sampling method for the field test. RESULTS: The interview version and the self-administered version was found equivalent. The ICC values of the domain scores ranged from 0.73 in the environment domain to 0.83 in the psychological domain. The face-to-face interview and telephone interview mode of administration were also found equivalent. The ICC for the domain scores ranged from 0.76 in the social interaction domain to 0.84 in the psychological domain. The other psychometric properties of the interview version were found comparable to the self-administered version. CONCLUSION: The self-administered and the interview version of the WHOQOL-BREF are regarded as identical in group comparison. The authors advise that it is acceptable to use different versions on different subjects in the same study, provided that the same version is applied on the same subject throughout the study. PMID- 16047517 TI - Validation of the chronic heart failure questionnaire (Chinese version). AB - A striking rise in the incidence and prevalence of congestive heart failure (CHF) in the Chinese population has been reported. While promoting the quality of life (QOL) of these patients is widely recognized as an important clinical priority, there is not a validated Chinese disease-specific QOL measure for CHF patients. The aim of this study was to validate the Chinese version of Chronic Heart Failure Questionnaire (CHQ-C) which measures the disease-specific QOL of patients with CHF. A sample of 110 CHF elderly patients and a comparison group of 64 healthy elderly people was recruited. Structured questionnaires were administered by a research nurse. Criterion and construct validity of CHQ-C was demonstrated by its significant correlation with the New York Heart Association classification (Spearman r = -0.52, p < 0.001) and the Chinese version of Hospital Anxiety and Depression scale (Pearson r = -0.74, p < 0.001), respectively. Known-groups comparison also demonstrated the ability of CHQ-C to differentiate the QOL of people with or without CHF. Confirmatory factor analysis supported the original four-factor structure of CHQ. In terms of internal consistency and test-retest reliability, CHQ-C demonstrated a Cronbach's alpha of 0.95 and an intraclass correlation coefficient (ICC) of 0.75, respectively. The responsiveness coefficient of CHQ-C was 0.63. In conclusion, CHQ-C is recommended as a valid and reliable tool for evaluating the QOL of Chinese patients with CHF. PMID- 16047518 TI - Reliability and validity of the Functional Assessment of Cancer Therapy General (FACT-G) in French cancer patients. AB - This report describes the reliability and validity of a French version of the Functional Assessment of Cancer Therapy - General (FACT-G) with a French sample of 493 cancer patients. The FACT-G consists of 27 items and four subscales: Physical (PWB), Functional (FWB), Social/Family (SFWB) and Emotional well-being (EWB). The study sample includes 64% with localized disease, 26% with metastases, 11% in remission, and 71% receiving radiation/chemotherapy. Internal consistency Cronbach alphas of the global FACT-G scale (0.90) and subscales (>0.75) are satisfactory (n = 126). Test-retest reproducibility is satisfactory for all subscales and the global scale (n = 87 to 93, r = 0.74 to 0.90). ANOVA models show that PWB differentiated between the three disease stages; the global FACT-G and FWB discriminated between patients with metastases and others with localized disease or in remission; EWB only discriminated between metastases and localized disease; while SFWB did not discriminate between groups at different stages of cancer. Only the PWB subscale discriminated between patients with no history from those receiving chemotherapy (p < or = 0.05). None of the scales discriminated between groups based on radiotherapy. These results may be useful in the design and interpretation of clinical trials involving French patients when the FACT-G is the outcome measure. PMID- 16047519 TI - Validating and norming of the Greek SF-36 Health Survey. AB - The main objective of this study was to validate the Greek SF-36 Health Survey and to provide general population normative data. The survey was administered to a stratified representative sample (n = 1426) of the general population residing in the broader Athens area and the response rate was 70.6%. Statistical analysis, according to documented procedures developed within the IQOLA Project, was performed. The missing value rate was very low, ranging from 0.1 to 1.3% at the item level. Multitrait scaling analysis confirmed the hypothesized scale structure of the SF-36. Cronbach's alpha coefficient met the criterion (>0.70) for group analysis in all eight scales. Known group comparisons yielded consistent support of construct validity of the SF-36. Significant statistical differences in mean scores were observed in relation to demographic and social characteristics such as gender, age, education and marital status. PMID- 16047520 TI - Adolescent-to-young adulthood heavy drinking trajectories and their prospective predictors. AB - OBJECTIVE: The purpose of this study was to investigate longitudinal trajectories of heavy drinking for males and females from adolescence to young adulthood, across the age span of 16-25 years, and to identify prospective predictors of the trajectory groups identified. METHOD: This study used semiparametric group-based mixture modeling to derive adolescent to young adult longitudinal trajectories of heavy drinking separately for 760 participants (430 females and 330 males) who have been participating in a long-term prospective study of risk factors for the development of heavy drinking and alcohol disorders. RESULTS: Four trajectory groups were identified for males and five for females; the trajectories indicated both continuity and change in heavy drinking across time for the trajectory groups identified. Major common prospective predictors for the high and very high heavy drinking trajectory groups supported the influences of values and beliefs (e.g., religious commitment), stressful life events and substance use. Additional predictors for males included lower academic functioning and task orientation, and for females, more frequent sexual behavior and general deviance. CONCLUSIONS: In this predominantly white, middle-class sample, we identified groups of frequent, heavy drinking teens during the middle-adolescent years. Our findings suggest that the frequency of heavy drinking behavior will further increase for some teens into their young adult years. The potential adverse consequences of heavy drinking among adolescents and young adults suggests that multitargeted, gender-specific, early interventions with these high-risk teens is important. PMID- 16047521 TI - Examining personality and alcohol expectancies using functional magnetic resonance imaging (fMRI) with adolescents. AB - OBJECTIVE: Personality and alcohol expectancies have been examined as risk factors for the initiation and maintenance of alcohol use in adolescents and young adults. Differences in processing appetitive stimuli are seen as a mechanism for personality's influence on behavior, and that mechanism predisposes individuals to form more positive expectancies for alcohol. The go/no-go task has been used to show how personality differences influence responding to appetitive stimuli in adolescents and adults, and functional magnetic resonance imaging (fMRI) has been used to examine the relation of go/no-go responding to personality in adult males. However, no study to date has examined the relation between fMRI responding, personality and alcohol expectancies in adolescents. METHOD: Forty-six adolescents (ages 12-14 years; 61% male) with minimal substance use histories completed measures of neuroticism, extraversion, and alcohol expectancies, and performed a go/no-go task during fMRI acquisition. RESULTS: Greater blood oxygen level-dependent (BOLD) response to inhibition predicted fewer expectancies of cognitive and motor improvements but more expectancies of cognitive and motor impairment from alcohol. In addition, extraverted youths reported more positive alcohol expectancies. However, BOLD response did not predict neuroticism or extraversion. CONCLUSIONS: These preliminary results suggest that decreased inhibitory neural processing may contribute to more positive and less negative expectancies, which can eventually lead to problem drinking. Further, extraversion may also yield more positive expectancies and could underlie a vulnerability to disordered alcohol use. PMID- 16047522 TI - Cue reactivity in adolescents: measurement of separate approach and avoidance reactions. AB - OBJECTIVE: There were two specific goals for the current study: (1) to demonstrate that adolescents display drug-specific cue reactivity to alcohol and cigarette visual cues that varies based on drug-use history and (2) to test the unique contribution of adolescents' avoidance reactions to alcohol and cigarette cues, independent of approach/craving reaction. METHOD: Adolescents (N = 143; age 13-20 years; 58 males) with varied substance-use histories were recruited from school and community sites. Adolescents were presented with a series of alcohol, cigarette, and nondrug comparison visual cues and reported their approach/craving and avoidance reactions. They also completed individual difference measures related to their alcohol and cigarette use and experiences. RESULTS: When adolescents were grouped according to their current alcohol or cigarette use (no use, low use, high use), increased use of alcohol or cigarettes was associated with stronger reactions (increased approach, decreased avoidance) to cues for that substance but not to nondrug control cues. Simultaneous regression analyses demonstrated that after controlling for approach/craving reactions, avoidance cue reactions predicted unique and/or incremental variance in measures of alcohol and cigarette usage, recent change in patterns of use, alcohol expectancies, alcohol restraint and parental alcohol problems. CONCLUSIONS: Adolescents displayed robust alcohol and cigarette cue-specific reactions that varied systematically with their current use of these drugs. Across numerous clinically relevant individual difference variables, predictive power was greatly enhanced through the inclusion of both avoidance and approach reactions. PMID- 16047523 TI - Coping motives as a moderator of daily mood-drinking covariation. AB - OBJECTIVE: The current study examines the extent to which college students' reports of drinking to cope (DTC) with negative affect moderate the daily covariation between specific types of negative mood (sadness, fear, hostility, shyness and boredom) and alcohol use. METHOD: Participants were full-time college students, aged 18-20, attending a large Southeastern university (N = 72; 50% male). These individuals completed an experience-sampling protocol over a 1-month interval, to assess daily mood and alcohol use. RESULTS: A series of Hierarchical General Linear Models found that individuals who reported low motives to cope through drinking showed an expected absence of daily mood and drinking covariation. For those reporting high coping motives, a complex and somewhat counterintuitive series of findings were found; students high in DTC drank less on days in which they experienced greater sadness. Analyses on the quadratic effects of mood revealed that when experiencing moderate to high levels of fear and shyness, individuals high in DTC were more likely to drink. For those low in coping motivations, fear and shyness did not predict daily drinking. CONCLUSIONS: Findings highlight the need to better our understanding of the exact meaning of DTC measures. In addition, there is a need to study how the context of college drinking influences self-medication and students' self-reports of DTC. PMID- 16047524 TI - Alcohol cue reactivity in alcohol-dependent adolescents. AB - OBJECTIVE: There is substantial evidence that adults with alcohol dependence show different responses (increased craving, increased salivation, changes in heart rate) to alcohol-related stimuli (i.e., alcohol cue reactivity) than nonalcoholics. Alcohol cue reactivity appears to be related to dependence severity and has been used to predict treatment outcomes, where more reactive alcoholics have poorer outcomes than less reactive alcoholics. Adolescents may also develop alcohol dependence, though it is uncertain whether they experience craving and cue reactivity in the presence of alcohol-associated stimuli. METHOD: To examine whether adolescents with alcohol dependence show alcohol cue reactivity, 28 alcoholic adolescents and 25 nonalcoholic adolescents (ages 14-19 years) were compared using a standard cue reactivity procedure, where participants view, hold and sniff different beverages, one of which is their preferred alcoholic beverage. Cue reactivity was assessed with subjective craving ratings, salivation (grams), and heart rate (beats per minute). Analyses were conducted by covarying response to control beverages. RESULTS: Alcoholics responded with both greater craving and greater salivation to the alcohol cue (controlling for response to control cues) than did nonalcoholics, supporting the hypothesis that adolescent alcoholics show alcohol cue reactivity. Heart rate showed no differential cue effect between alcoholics and controls. CONCLUSIONS: Findings support that adolescent alcoholics experience alcohol cue reactivity, as evidenced by increased salivation and subjective craving in the presence of alcohol-related stimuli. Investigation of treatments that may reduce alcohol craving and cue reactivity in adolescents with alcohol dependence is warranted. PMID- 16047525 TI - Alcohol screening and brief intervention in primary care settings: implementation models and predictors. AB - OBJECTIVE: This study compared two different implementation strategies for Cutting Back, a primary care alcohol screening and brief intervention (SBI) program for hazardous and harmful drinkers. It also identified organizational factors contributing to the success or failure of SBI implementation. METHOD: Cutting Back was implemented in 10 primary care practices associated with managed care organizations (MCOs) in five states, through a system of planning, training, technical assistance and clinic feedback. Clinics were randomly assigned to one of two brief intervention systems: In the P Model, medical providers were responsible for delivering interventions, whereas in the S Model mid-level professionals (usually nurses) acted as the clinic specialists to provide that service. Data were collected to measure the performance of screening and delivery of interventions in each clinic. RESULTS: The S Model screened a higher percentage of patients than did the P Model during the best month of program operation (50% vs 44%) and over all months of operation (24% vs 19%). Of those patients who screened positive, more patients in the S condition received an intervention than in the P condition (73.1% vs 57.1%), but there was a considerable range of performance among the five sites within each condition. Results at the clinic level were mixed, with some MCOs performing alcohol SBI significantly better with the S model and others doing better with the P model. The ability of clinics to conduct SBI was significantly correlated with both provider characteristics and organizational factors (e.g., prior SBI experience, MCO stability, number of clinicians trained and the quality of the MCO coordinator's work). Lack of provider time, staff turnover and competing priorities correlated negatively with SBI performance. CONCLUSIONS: The extent to which a given delivery model is likely to work best within an MCO depends on complex provider and organizational characteristics. PMID- 16047526 TI - Is alcohol assessment therapeutic? Pretreatment change in drinking among alcohol dependent women. AB - OBJECTIVE: Participants in alcohol treatment outcome studies typically undergo extensive assessment protocols to determine eligibility and provide descriptive and predictive data before beginning therapy. Changes in alcohol consumption as a result of reactivity to assessment have generally not been studied; most researchers collapse data across the pretreatment period and compare them with within-treatment and posttreatment periods. Previously we found a reduction in drinking days from 90 days prebaseline to 3 months postbaseline, with no significant additional reduction during the second 3 months of treatment, in a clinical trial of cognitive-behavioral therapy (CBT) for alcohol-dependent women. The current study examined the points at which these participants stopped or reduced their drinking during the pretreatment assessment period. METHOD: As part of a randomized clinical trial of 6 months of individual or couples CBT for 102 alcoholic women, study participants were assessed briefly via a telephone screen, a 90-minute clinical intake session with their spouses, and then a 3- to 4-hour baseline research interview. RESULTS: Changes in drinking frequency occurred at all four points in the pretreatment assessment process, resulting in 44% of the participants becoming abstinent before the first session of treatment. A decrease in drinking quantity across the assessment period also was found. Participants who significantly reduced drinking prior to Session 1 demonstrated better drinking outcomes during and 12 months after treatment. CONCLUSIONS: Changes in alcohol consumption during the assessment process in treatment outcome studies should be examined in future studies, as implications abound for interpretation of results from clinical trials. Also, implications regarding the active ingredients in brief interventions and mechanisms of the therapeutic impact of alcohol assessment should be further addressed. PMID- 16047527 TI - A multicountry controlled trial of strategies to promote dissemination and implementation of brief alcohol intervention in primary health care: findings of a World Health Organization collaborative study. AB - OBJECTIVE: This study examines the impact of marketing strategies on the dissemination of a brief alcohol intervention program to general practitioners (GPs). The marketing strategy was tested to determine the most effective way to promote awareness about and consideration of a brief alcohol intervention program. The study also examines the impact of training and support strategies to promote the program's implementation in routine primary care. METHOD: A pragmatic trial was carried out in Australia, Belgium (Flanders), Denmark, England, New Zealand and Spain (Catalonia) in which GPs were randomly allocated into one of three marketing conditions (direct mail, telemarketing and academic detailing [personal visits]). The GPs who requested a brief intervention program and agreed to use it were stratified by previous marketing condition and randomly allocated into one of three implementation strategy groups: written guidance, outreach training and outreach training plus ongoing telephone support. RESULTS: Acceptance of the brief intervention program was more effective with use of telemarketing (65%) and academic detailing (67%) than with direct mail (32%) for promoting awareness about and consideration of a brief alcohol intervention program. The median proportion of patients screened was higher for trained GPs (6%) and supported GPs (9%) than for control GPs (1%), who received only written guidance on how to conduct brief intervention. Similarly, the median rate for giving advice to at-risk patients was higher for trained GPs (3%) and supported GPs (3%) than for control GPs (0%). CONCLUSIONS: The adoption of more direct approaches for disseminating evidence-based intervention programs to GPs is a necessary first step for changing practice behavior. However, outreach training was required to promote actual use of a new procedure in routine practice. PMID- 16047528 TI - Brief physician advice for problem drinking among older adults: an economic analysis of costs and benefits. AB - OBJECTIVE: Problem alcohol use among elderly persons can have a variety of health related consequences, complicating management of chronic illnesses and increasing health care utilization and costs. This study evaluates the economic cost and benefits of brief intervention for at-risk drinking older adults. METHOD: A controlled clinical trial with 24-month follow-up tested effectiveness of brief physician advice in reducing alcohol use, health care utilization and other consequences among older (age 65 or older) adult problem drinkers. Of 6,073 patients screened for problem drinking in 24 community-based primary care practices in Wisconsin, 158 patients met inclusion criteria and were randomized into control (n = 71) or intervention (n = 87) groups. Intervention group patients received two 10- to 15-minute physician-delivered counseling sessions including professional advice, education and contracting using scripted workbooks. RESULTS: The intervention group demonstrated significant reductions in alcohol use (p = 0.001) and frequency of excessive drinking (p = 0.03) compared with the control group over 24 months, but no significant differences emerged in economic outcomes, including hospital days, emergency department visits, office visits, medications, lab and x-ray procedures, injuries, legal events or mortality. CONCLUSIONS: Although the clinical benefits of brief alcohol interventions with older adults are clear, the economic results in this age group are less certain. Older adult problem drinkers may require more intensive and costly interventions to achieve economic benefits similar to those seen in younger adult problem drinkers. Methodological issues, such as statistical power, outcome measures, outlier cases and follow-up periods, are identified for future evaluations. PMID- 16047529 TI - Functioning, problem behavior and health services use among nursing home residents with alcohol-use disorders: nationwide data from the VA minimum data set. AB - OBJECTIVE: We examined (1) whether nursing home residents with alcohol-use disorders (AUDs) function more poorly, have more behavioral problems and use more health services than do demographically matched controls without such disorders, and (2) whether AUDs interact with alcohol consumption to predict poorer behavioral and health services outcomes among nursing home residents. METHOD: We used Department of Veterans Affairs (VA) Patient Treatment File data to identify a nationwide sample of older VA nursing home residents with recent AUD diagnoses (n = 3,336) and a demographically matched sample without such diagnoses (n = 3,336). The groups were compared on the Resident Assessment Instrument Minimum Data Set indices of health-related functioning, substance use, problem behavior and health services use. We conducted two-way analyses of variance to determine interactions between presence of an AUD and alcohol consumption on problem behavior and health services use. RESULTS: Residents with AUD diagnoses functioned somewhat better than demographically matched controls. However, they were more likely to have falls and fractures, difficulties in social functioning and to use more health services. AUD diagnoses interacted with alcohol consumption to predict an elevated risk of falls and fractures and more mental health services use. CONCLUSIONS: Older residents with an AUD form a distinct nursing home population that functions somewhat better than residents without such disorders but they may pose more challenges to staff and use more health care services. Residents' drinking histories should be considered in formulating nursing home policies about alcohol consumption. PMID- 16047530 TI - Initial depression and subsequent drinking during alcoholism treatment. AB - OBJECTIVE: Individuals entering treatment for alcoholism have elevated depressive symptoms that in turn may affect response to treatment, including drinking outcomes. The purpose of the study is to examine the impact of depression at treatment entry on drinking over the course of treatment. METHOD: The Project MATCH (Matching Alcoholism Treatment to Client Heterogeneity) data set, a randomized, multisite psychosocial treatment trial for alcoholism, was analyzed. The sample consisted of 1,450 subjects, of whom 1,102 (76.0%) were male and 348 (24.0%) were female. Cross-lagged analyses of (1) depression and drinking intensity and (2) depression and drinking frequency were conducted using path analysis. Covariates were age, gender, race, and treatment assignment. Analyses focused on the 3-month active treatment phase of the trial. RESULTS: Depression at treatment entry predicted more intense drinking and more frequent drinking, respectively, in the first month of treatment but showed little association with drinking in Months 2 and 3. CONCLUSIONS: Individuals entering treatment for alcoholism with elevated levels of depression may be slower to benefit from treatment. Because the initial phase of treatment may be crucial to successful engagement and retention, the development of interventions to improve early success in treatment among individuals with elevated levels of depression may be beneficial. Future studies should examine the long-term, bidirectional relationship of depression and drinking following treatment. PMID- 16047531 TI - Is there a relationship between victim and partner alcohol use during an intimate partner violence event? Findings from an urban emergency department study of abused women. AB - OBJECTIVE: This study sought to identify factors associated with drinking during an intimate partner violence (IPV) event among abused women presenting to an urban emergency department (ED). METHODS: We use a cross-sectional study of IPV cases among adult female patients seen at an urban ED. Bivariate and logistic regression analyses were performed to identify substance use factors associated with an abused woman drinking while victimized or perpetrating IPV. RESULTS: Among the 182 cases, an increased number of drinks per week, consuming five or more drinks per occasion, alcohol abuse and dependence, and illicit drug use were significantly associated with the abused woman's drinking while victimized or perpetrating IPV Partner's drinking five or more drinks per occasion was associated only with the woman's drinking while victimized. Partners were more likely to drink while perpetrating IPV in the relationship whether or not the woman drank while victimized. Among couples in which the abused woman also perpetrated violence, the partner's drinking more closely paralleled the woman's drinking in events perpetrated by the woman. Independent risk factors associated with the abused woman drinking during victimization included number of drinks she consumed per week (adjusted odds ratio [adj. OR] = 1.31 for every five drinks) and her illicit drug use (adj. OR = 4.3). The odds of an abused woman drinking while perpetrating IPV increased 1.4 times for every five drinks she consumed per week. CONCLUSIONS: These findings suggest that alcohol-related behavior by both couples and individuals are important factors to consider in the relationship between IPV and alcohol use in this population. PMID- 16047532 TI - A further examination of gender differences in alcohol-related aggression. AB - OBJECTIVE: The purpose of this investigation was to replicate and extend findings from a previous study on the acute effects of alcohol on aggressive behavior in men and women in a laboratory setting. METHOD: Subjects were 234 (111 men and 123 women) healthy social drinkers between 21 and 35 years of age. They were randomly assigned to either an alcohol or a placebo group. Aggression was measured using a modified version of the Taylor Aggression Paradigm, in which electric shocks are received from and administered to a fictitious opponent during a supposed competitive interpersonal task. Aggression was operationalized as the intensity and duration of shocks that subjects administered to their "opponent." RESULTS: Provocation was a stronger elicitor of aggression than either gender or alcohol. Overall, alcohol increased aggression for men but not for women. CONCLUSIONS: In conjunction with other laboratory investigations on alcohol-related aggression, this study suggests that alcohol increases aggression for men but not for women. This finding may be due to gender-related differences in liability thresholds for aggression as well as discrepancies in how men and women respond to different forms of provocation. PMID- 16047533 TI - Predictors of completion status in a remedial program for male convicted drinking drivers. AB - OBJECTIVE: Rates of attrition in alcohol and drug treatment programs are often greater than 50%, and completion of treatment has been shown to be a potent predictor of posttreatment outcome. The current study examined both rates and predictors of completion among male participants in a remedial measures program for convicted drinking drivers. METHOD: Male individuals (n = 5,409) convicted of a drinking driving offense in Ontario between October 2000 and December 2002 who did and did not complete a mandatory rehabilitation program were described in terms of demographic, drug use and legal variables collected at time of assessment. RESULTS: The program completion rate was extremely high (97.3%). In multivariate analyses, noncompleters-relative to completers-were younger; drank more frequently; were less likely to own a home; and were more likely to live in urban centers, have two or more lifetime impaired driving convictions and have experienced more than one adverse consequence of substance use. CONCLUSIONS: Ontario's remedial measures program for convicted drinking drivers, in which the return of a suspended license after the period of mandatory suspension is contingent on the completion of the program, demonstrates a very low level of client attrition. Individuals who do not complete the program bear many similarities to those at high risk for persistent drink-drive behavior and its associated negative health consequences. PMID- 16047534 TI - Cross-national performance of the RAPS4/RAPS4-QF for tolerance and heavy drinking: data from 13 countries. AB - OBJECTIVE: There are little data available on the performance of brief screening instruments for alcohol-use disorders cross-nationally; therefore, we analyzed the performance of one such instrument in a number of countries. METHOD: Performance of the RAPS4 for tolerance and the RAPS4-QF for heavy drinking are analyzed from emergency room data across 13 countries included in the combined Emergency Room Collaborative Alcohol Analysis Project (ERCAAP) and the World Health Organization Collaborative Study on Alcohol and Injuries. RESULTS: The RAPS4 showed good sensitivity and specificity for tolerance across most of the countries, but was higher in countries that were higher on societal-level detrimental drinking patterns. Prevalence of tolerance was also higher in those countries with high detrimental drinking pattern scores. Sensitivity of the RAPS4 QF for heavy drinking was uniformly high across countries, while maintaining good specificity, and did not vary by detrimental drinking patterns. CONCLUSIONS: Findings suggest the RAPS4 and RAPS4-QF may hold promise cross-nationally. Future research should more fully address the performance of brief screening instruments for alcohol-use disorders (using standard diagnostic criteria) cross-nationally, with consideration of the impact of societal drinking patterns. PMID- 16047535 TI - Can the short index of problems (SIP) be improved? Validity and reliability of the three-month SIP in an emergency department sample. AB - OBJECTIVE: Although the Short Index of Problems (SIP) is often used, little is known about the psychometric properties of the SIP in special populations. The present study seeks to determine the following: (1) whether it is possible to substitute items to enhance the psychometric properties of the SIP and (2) whether the SIP, or improved scale, is as sensitive as the Drinker Inventory of Consequences (DrInC) to assess intervention effectiveness. METHOD: The sample consisted of 404 injured patients who were treated in the Emergency Department (ED) of a major teaching hospital that serves southern New England. Three approaches were used to guide development of the 3-month SIP-R, the potential alternative to the SIP. Cronbach's alpha assessed intrascale reliability; hierarchical multiple regression assessed construct validity; performance of the scales assessing intervention change were compared to the total 3-month DrInC as a function of intervention using analysis of covariance (ANCOVA). RESULTS: There was no evidence that changing the current SIP items will significantly improve performance. The 3-month SIP performed as well as the 3-month DrInC-2R in predicting 12-month DrInC scores and in determining intervention change at 12 months. Of the 45 DrInC items, 31 also predicted a difference across intervention groups. CONCLUSIONS: These results suggest there is no advantage to changing the current SIP items. The 3-month SIP is a psychometrically sound measure for assessing consequences of alcohol consumption in an ED sample and is almost as sensitive to intervention change as the full DrInC. PMID- 16047536 TI - "Binge" drinking and blood alcohol concentration. PMID- 16047537 TI - "Binge" drinking and blood alcohol concentration. PMID- 16047538 TI - Is alcohol consumption a risk factor for weight gain and obesity? AB - Alcohol represents an important source of energy. Despite its comparatively high energy content of 7.1 g/kcal, it is still controversial whether moderate amounts of alcohol represent a risk factor for weight gain and obesity. Epidemiologic data showed a positive, negative, or no relationship between alcohol intake and body weight. Despite the difficulty in assessing alcohol intake as well as controlling for different confounders of the energy-balance equation, the conflicting epidemiologic data can be explained in most instances. Every component of the energy-balance equation is affected by the ingestion of alcohol. Moderate amounts of alcohol enhance energy intake due to the caloric content of the alcohol as well as its appetite-enhancing effects. Alcohol-induced thermogenesis is approximately 20% in healthy nonalcoholic subjects, i.e., moderate alcohol consumers, which is higher than for other energy substrates but considerably lower than in heavy alcohol consumers. This would suggest that a major fraction of the alcohol energy represents a navailable energy source for ATP synthesis in moderate non-daily alcohol consumers. Experimental evidence from several metabolic studies showed a suppression of lipid oxidation by alcohol and thus the enhancement of a positive fat balance. The nonoxidized fat is preferentially deposited in the abdominal area. The experimental metabolic evidence suggests that the consumption of moderate amounts of alcohol has to be accounted for in the energy-balance equation and may represent a risk factor for the development of a positive energy balance and thus weight gain. In the heavy alcohol consumer and eventually also in daily moderate alcohol consumers, a larger fraction of the alcohol energy might not be an available source of energy due to the induction of the microsomal ethanol-oxidizing system (MEOS). Experimental data in combination with epidemiologic findings suggest that alcohol energy counts more in moderate nondaily alcohol consumers than in some moderate daily and all heavy consumers. Accordingly the question is not "Whether alcohol calories do count" but "How much do alcohol calories count?". There seems to be a large individual variability according to the absolute amount of alcohol consumed, the drinking frequency as well as genetic factors. Presently it can be said that alcohol calories count more in moderate nondaily consumers than in daily (heavy) consumers. Further, they count more in combination with a high-fat diet and in overweight and obese subjects. PMID- 16047539 TI - Bone disease in primary hypercalciuria. AB - Primary hypercalciuria (PH) is very often accompanied by some degree of bone demineralization. The most frequent clinical condition in which this association has been observed is calcium nephrolithiasis. In patients affected by this disorder, bone density is very frequently low, and increased susceptibility to fragility fractures is reported. The very poor definition of this bone disease from a histomorphometric point of view is a crucial aspect. At present, the most common finding seems to be a low bone turnover condition. Many factors are involved in the complex relationships between bone loss and PH. Since bone loss was mainly reported in patients with fasting hypercalciuria, a primary alteration in bone metabolism was proposed as a cause of both hypercalciuria and bone demineralization. This hypothesis was strengthened by the observation that some bone resorbing-cytokines, such as interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor nechrosis factor-alpha (TNF-alpha), are high in hypercalciuric patients. An excessive response to the acid load induced by dietary protein intake seems to be an additional factor explaining a primitive alteration of bone. The intestine plays a major role in the clinical course of bone disease in PH. Patients with absorptive hypercalciuria less frequently show bone disease, and a reduction in dietary calcium greatly increases the probability of bone loss in PH subjects. It has recently been reported that greater bone loss is associated with a larger increase in intestinal calcium absorption in PH patients. Considering the absence of parathyroid hormone (PTH) alterations, it was proposed that this is not a compensatory phenomenon, but probably the marker of disturbed cell calcium transport, involving both intestinal and bone tissues. While renal hypercalciuria is rather uncommon, the kidney still seems to play a role in the pathogenesis of bone loss in PH patients, possibly via the effect of mild-to-moderate urinary phosphate loss with secondary hypophosphatemia. In conclusion, bone loss is very common in PH patients. Even if most of the factors involved in this process have been identified, many aspects of this intriguing clinical condition remain to be elucidated. PMID- 16047540 TI - Molecular aspects of thrombosis and antithrombotic drugs. AB - There have been major advances in our understanding of thrombosis and antithrombotic drugs. This review focuses on the molecular aspects of thrombus formation and antithrombotic therapy. Molecules involved in arterial thrombosis are derived from inflammatory cells in the atherosclerotic plaque and blood platelets. These molecules work in concert to promote plaque instability and thrombogenicity. Thrombus formation on the ruptured plaque is mediated by platelet and coagulation activation. By contrast, molecules involved in venous thrombosis are derived from the activated coagulation cascade. Platelets appear to play a secondary role. The antithrombotic drugs are classified according to their targeted constituents: antiplatelet agents and anticoagulants; the latter are further divided into non-specific anticoagulants, such as vitamin K antagonists and heparin, and direct thrombin inhibitors, including hirudin and argatroban. Currently available antiplatelet agents target glycoprotein IIbIIIa (abciximab, tirofiban, eptifibatide), cyclooxygenase-1 (aspirin) or adenosine diphosphate receptor, P2Y12 (clopidogrel). PMID- 16047541 TI - In praise of "natural history". PMID- 16047542 TI - Synaptically activated ca2+ release from internal stores in CNS neurons. AB - Synaptically activated postsynaptic [Ca2+]i increases occur through three main pathways: Ca2+ entry through voltage-gated Ca2+ channels, Ca2+ entry through ligand-gated channels, and Ca2+ release from internal stores. The first two pathways have been studied intensively; release from stores has been the subject of more recent investigations. Ca2+ release from stores in CNS neurons primarily occurs as a result of IP3 mobilized by activation of metabotropic glutamatergic and/or cholingergic receptors coupled to PLC. Ca2+ release is localized near spines in Purkinje cells and occurs as a wave in the primary apical dendrites of pyramidal cells in the hippocampus and cortex. The amplitude of the [Ca2+]i increase can reach several micromolar, significantly larger than the increase due to backpropagating spikes. The large amplitude, long duration, and unique location of the [Ca2+]i increases due to Ca2+ release from stores suggests that these increases can affect specific downstream signaling mechanisms in neurons. PMID- 16047544 TI - Electrical potentials indicate stimulus expectancy in the brains of ants and bees. AB - In vertebrates, and in humans in particular, so-called 'omitted stimulus potentials' can be electrically recorded from the brain or scalp upon repeated stimulation with simple stimuli such as light flashes. While standard evoked potentials follow each stimulus in a series, 'omitted stimulus potentials' occur when an additional stimulus is expected after the end of a stimulus series. These potentials represent neuronal plasticity and are assumed to be involved in basic cognitive processes. We recorded electroretinograms from the eyes and visually evoked potentials from central brain areas of honey bees and ants, social insects to which cognitive abilities have been ascribed and whose rich-behavioral repertoires include navigation, learning and memory. We demonstrate that omitted stimulus potentials occur in these insects. Omitted stimulus potentials in bees and ants show similar temporal characteristics to those found in crayfish and vertebrates, suggesting that common mechanisms may underlie this form of short term neuronal plasticity. PMID- 16047543 TI - Synaptic and extrasynaptic secretion of serotonin. AB - Serotonin is a major modulator of behavior in vertebrates and invertebrates and deficiencies in the serotonergic system account for several behavioral disorders in humans. The small numbers of serotonergic central neurons of vertebrates and invertebrates produce their effects by use of two modes of secretion: from synaptic terminals, acting locally in "hard wired" circuits, and from extrasynaptic axonal and somatodendritic release sites in the absence of postsynaptic targets, producing paracrine effects. In this paper, we review the evidence of synaptic and extrasynaptic release of serotonin and the mechanisms underlying each secretion mode by combining evidence from vertebrates and invertebrates. Particular emphasis is given to somatic secretion of serotonin by central neurons. Most of the mechanisms of serotonin release have been elucidated in cultured synapses made by Retzius neurons from the central nervous system of the leech. Serotonin release from synaptic terminals occurs from clear and dense core vesicles at active zones upon depolarization. In general, synaptic serotonin release is similar to release of acetylcholine in the neuromuscular junction. The soma of Retzius neurons releases serotonin from clusters of dense core vesicles in the absence of active zones. This type of secretion is dependent of the stimulation frequency, on L-type calcium channel activation and on calcium induced calcium release. The characteristics of somatic secretion of serotonin in Retzius neurons are similar to those of somatic secretion of dopamine and peptides by other neuron types. In general, somatic secretion by neurons is different from transmitter release from clear vesicles at synapses and similar to secretion by excitable endocrine cells. PMID- 16047545 TI - Changes in heart rate associated with contest outcome in agonistic encounters in lobsters. AB - Agonistic contests between lobsters housed together in a confined space progress through encounters of increasing intensity until a dominance relationship is established. Once this relationship is established, losing animals continually retreat from the advances of winners. These encounters are likely to consume much energy in both winning and losing animals. Therefore, one might expect involvement of many physiological systems before, during and after fights. Here, we report effects of agonistic encounters on cardiac frequency in winning and losing adult lobsters involved in dyadic interactions. The results show that: (i) small but significant increases in heart rate are observed upon chemical detection of a conspecific; (ii) during agonistic interactions, further increases in heart rate are seen; and (iii) ultimate winners exhibit greater increases in heart rate lasting longer periods of time compared to ultimate losers. Heart rate in winners remains elevated for at least 15 min after the contests have ended and animals have been returned to their home tanks. Reduced effects are seen in second and third pairings between familiar opponents. The sustained changes in heart rate that we observe in winning lobsters may result from hormonal modulation of cardiac function related to the change in social status brought about by contest outcome. PMID- 16047546 TI - Dopaminergic modulation of neurosecretory cells in the crayfish. AB - The main aims of this paper are (a) to locate possible dopaminergic neurons in the eyestalk with anti-tyrosine hydroxylase antibodies, (b) to search for the presence of dopamine (DA) in the nervous structures of the eyestalk, (c) to explore its release, and (d) to test the effect of DA on neurosecretory cells in the eyestalk. Experiments were performed in adult crayfishes Procambarus clarkii, in isolated optic peduncle. Immunocytochemistry was made with the antibody against its precursor synthesizing enzyme tyrosine-hydroxylase. The content and release studies of DA were made using high performance liquid chromatography (HPLC). Extracellular and intracellular recordings were conducted with conventional recording techniques. A large number (approximately 2000) of immunopositive somata of different sizes and shapes were identified in various regions of the eyestalk. The majority of somata are of the smallest size (5-25 microm diameter). DA content in the eyestalk was 5.6 +/- 0.1 pmol per structure; the greatest content is in the MT (over 60%). A basal level release of DA was observed. Incubation of eyestalks in solution containing a high K+ concentration increased the DA release (79%). Two effects of DA on the excitability of X-organ neurons were observed; an excitatory effect on neurons of approximately 25 microm somata diameter and another inhibitory effect in the group of approximately 35 microm somata diameter neurons. The excitation occurs with a depolarization and decrement of membrane conductance in the cell soma while the inhibition occurs with a hyperpolarization and increment of membrane conductance in soma. We concluded the following: (1) Dopamine is present in each optic ganglia of the crayfish eyestalk. (2) There is a basal release of DA from the isolated eyestalk. (3) DA release is enhanced threefold by eyestalk incubation in 40 mM [K+] solution. (4) DA selectively excites a population of neurons with low-speed conduction axons, and small somata in the X-organ-sinus gland system, while inhibiting another population characterized by higher axonal conduction speed and large somata. (5) These observations support a role for DA as a neurotransmitter or neuromodulator in the X-organ neurons of the crayfish eyestalk. PMID- 16047547 TI - Role of nitric oxide on motor behavior. AB - The present review paper describes results indicating the influence of nitric oxide (NO) on motor control. Our last studies showed that systemic injections of low doses of inhibitors of NO synthase (NOS), the enzyme responsible for NO formation, induce anxiolytic effects in the elevated plus maze whereas higher doses decrease maze exploration. Also, NOS inhibitors decrease locomotion and rearing in an open field arena. These results may involve motor effects of this compounds, since inhibitors of NOS, NG-nitro-L-arginine (L-NOARG), N(G)-nitro-L arginine methylester (L-NAME), N(G)-monomethyl-L-arginine (L-NMMA), and 7 Nitroindazole (7-NIO), induced catalepsy in mice. This effect was also found in rats after systemic, intracebroventricular or intrastriatal administration. Acute administration of L-NOARG has an additive cataleptic effect with haloperidol, a dopamine D2 antagonist. The catalepsy is also potentiated by WAY 100135 (5-HT1a receptor antagonist), ketanserin (5HT2a and alfal adrenergic receptor antagonist), and ritanserin (5-HT2a and 5HT2c receptor antagonist). Atropine sulfate and biperiden, antimuscarinic drugs, block L-NOARG-induced catalepsy in mice. L-NOARG subchronic administration in mice induces rapid tolerance (3 days) to its cataleptic effects. It also produces cross-tolerance to haloperidol induced catalepsy. After subchronic L-NOARG treatment there is an increase in the density NADPH-d positive neurons in the dorsal part of nucleus caudate-putamen, nucleus accumbens, and tegmental pedunculupontinus nucleus. In contrast, this treatment decreases NADPH-d neuronal number in the substantia nigra compacta. Considering these results we suggest that (i) NO may modulate motor behavior, probably by interfering with dopaminergic, serotonergic, and cholinergic neurotransmission in the striatum; (ii) Subchronic NO synthesis inhibition induces plastic changes in NO-producing neurons in brain areas related to motor control and causes cross-tolerance to the cataleptic effect of haloperidol, raising the possibility that such treatments could decrease motor side effects associated with antipsychotic medications. Finally, recent studies using experimental Parkinson's disease models suggest an interaction between NO system and neurodegenerative processes in the nigrostriatal pathway. It provides evidence of a protective role of NO. Together, our results indicate that NO may be a key participant on physiological and pathophysiological processes in the nigrostriatal system. PMID- 16047548 TI - Electrophysiological responses of nucleus tractus solitarius neurons to CCK and gastric distension in newborn lambs. AB - The effect of cervical vagus nerve stimulation, gastric distension and CCK-8S administration was studied on the activity of 120 neurons located in the nucleus tractus solitarius (NTS) of anesthetized newborn lambs. One hundred cells responded to the three different inputs. The distribution of the cells in the NTS was from 3 mm rostral to 3 mm caudal to the obex, the major responsive cells being located at the level of the obex. Neurons were either excited or inhibited by gastric distension and CCK-8S, and the responses to these two stimuli were always in the same direction. A small number of cells responded to gastric distension and CCK-8S but not to vagus nerve stimulation. Injection of the CCK-A receptor antagonist 2-NAP abolished both the responses to CCK-8S and to gastric distension. The results are consistent with the idea that CCK-8S acts directly on vagal mechanoreceptive endings in the gastric corpus close to duodenum. These results from lambs may reflect the pathway by which gastric distension and peripheral CCK-8S modulate NTS cells activity during colostrum ingestion, which could in turn activate structures related to learning and memory processes involved in the development of mother preference. PMID- 16047549 TI - Differential expression of genes at stages when regeneration can and cannot occur after injury to immature mammalian spinal cord. AB - Comprehensive screens were made for genes that change their expression during a brief critical period in development when neonatal mammalian central nervous system (CNS) loses its capacity to regenerate. In newly born opossums older than 12 days regeneration ceases to occur in the cervical spinal cord. It continues for 5 more days in lumbar regions. The mRNA's expressed in cords that do and do not regenerate were analyzed by polymerase chain reaction-based subtractive hybridization. The mRNAs extracted from cervical cords of animals aged 9 and 12 days were subtracted reciprocally, old from young and young from old. Additional subtractions were made between lumbar regions of 12 day-old cords (which can regenerate) and cervical regions (which cannot). Mini libraries of approximately 2000 opossum cDNA clones resulted from each subtraction. Many sequences were novel. Others that were expressed differentially were related to cell growth, proliferation, differentiation, motility, adhesion, cytoskeleton and extracellular matrix. A major task was to narrow the search and to eliminate genes that were not associated with regeneration. Clones from different subtractions were cross-hybridized. After those common to regenerating and nonregenerating cords were rejected, approximately 284 sequences of interest remained. Our results revealed novel sequences, as well as genes involved in transcription, cell signaling, myelin formation, growth cone motility, liver regeneration, and nucleic acid and protein management as the candidates important for neuroregeneration. For selected genes of potential interest for regeneration (for example cadherin, catenin, myelin basic protein), their temporal and spatial distributions and levels of expression in the CNS were measured by Northern blots, semiquantitative and real-time RT-PCR, and in situ hybridization. Our experiments set the stage for testing the efficacy of candidate genes in turning on or off the capacity for spinal cord regeneration. Opossum spinal cords in vitro provide a reliable and rapid assay for axon outgrowth and synapse formation. PMID- 16047550 TI - Hirudo medicinalis: a platform for investigating genes in neural repair. AB - We have used the nervous system of the medicinal leech as a preparation to study the molecular basis of neural repair. The leech central nervous system, unlike mammalian CNS, can regenerate to restore function, and contains identified nerve cells of known function and connectivity. We have constructed subtractive cDNA probes from whole and regenerating ganglia of the ventral nerve cord and have used these to screen a serotonergic Retzius neuron library. This identifies genes that are regulated as a result of axotomy, and are expressed by the Retzius cell. This approach identifies many genes, both novel and known. Many of the known genes identified have homologues in vertebrates, including man. For example, genes encoding thioredoxin (TRX), Rough Endoplasmic Reticulum Protein 1 (RER-1) and ATP synthase are upregulated at 24 h postinjury in leech nerve cord. To investigate the functional role of regulated genes in neuron regrowth we are using microinjection of antisense oligonucleotides in combination with horseradish peroxidase to knock down expression of a chosen gene and to assess regeneration in single neurons in 3-D ganglion culture. As an example of this approach we describe experiments to microinject antisense oligonucleotide to a leech isoform of the structural protein, Protein 4.1. Our approach thus identifies genes regulated at different times after injury that may underpin the intrinsic ability of leech neurons to survive damage, to initiate regrowth programs and to remake functional connections. It enables us to determine the time course of gene expression in the regenerating nerve cord, and to study the effects of gene knockdown in identified neurons regenerating in defined conditions in culture. PMID- 16047551 TI - Repair and regeneration of functional synaptic connections: cellular and molecular interactions in the leech. AB - A major problem for neuroscience has been to find a means to achieve reliable regeneration of synaptic connections following injury to the adult CNS. This problem has been solved by the leech, where identified neurons reconnect precisely with their usual targets following axotomy, re-establishing in the adult the connections formed during embryonic development. It cannot be assumed that once axons regenerate specific synapses, function will be restored. Recent work on the leech has shown following regeneration of the synapse between S interneurons, which are required for sensitization of reflexive shortening, a form of non-associative learning, the capacity for sensitization is delayed. The steps in repair of synaptic connections in the leech are reviewed, with the aim of understanding general mechanisms that promote successful repair. New results are presented regarding the signals that regulate microglial migration to lesions, a first step in the repair process. In particular, microglia up to 900 microm from the lesion respond within minutes by moving rapidly toward the injury, controlled in part by nitric oxide (NO), which is generated immediately at the lesion and acts via a soluble guanylate cyclase (sGC). The cGMP produced remains elevated for hours after injury. The relationship of microglial migration to axon outgrowth is discussed. PMID- 16047552 TI - Neuron-like differentiation of PC12 cells treated with media conditioned by either sciatic nerves, optic nerves, or Schwann cells. AB - In previous works we reported the finding of neurotrophic activity in a serum free Dulbecco's modified Eagle's medium conditioned by rat sciatic nerves, previously maintained in culture for 11 days. This medium produces rapid neuron like differentiation of cultured PC12 cells, as revealed by an increase in the size of the cell body and by the extension of short and/or long neurites by most of the cells. Neuregulin present in the conditioned medium was demonstrated to play a key role in the observed differentiation. In the present work, taking into consideration those latter results, the neurotrophic activity of conditioned media prepared with sciatic and optic nerves cultured during days 1-4 and 9-12 were studied. Evaluation of the trophic activities of those media revealed an opposite timing in the activities of sciatic and optic nerves conditioned media. The activity of the sciatic nerve was not observed in the 1-4-day period, increasing then up to the 9-12-day period. On the contrary, the optic nerve conditioned medium was active in the 1-4-day period, decreasing down to the 9-12 day period. These results led us to explore the contribution of the different cellular constituents of those nerves to their neurotrophic properties. As a first step in that direction we also investigated the neurotrophic activity of media conditioned during 12 days by cultured Schwann cells isolated from rat sciatic nerves. The Schwann cell conditioned media did produce a rapid differentiation of the PC12 cells similar to that caused by the sciatic nerve conditioned medium, though of a lower magnitude. Variations in the trophic activities of the conditioned media used in the present work is discussed taking into consideration the production of trophic and inhibitory factors by the peripheral and central glial cells. The role played by the optic nerve glia and myelin is being investigated at present. PMID- 16047553 TI - Using herbal medicine as a means of prevention experience during the SARS crisis. AB - At the peak of the SARS epidemic in Hong Kong, hospital workers were under high risks of contraction of the infection. Herbal preparations had been used historically in China to treat influenza-like diseases. During the SARS outbreak, herbal preparations had been used jointly with standard modern treatment in China. As a means to protect the at-risk hospital workers, an innovative herbal formula was created and consumed by 3160 of them in two weeks. During the two weeks, symptoms and adverse effects were close monitored; 37 of them had their serum checked for immunological responses. The results showed that none of the herb consumers contracted the infection, compared to 0.4% among the non consumers. Adverse effects had been infrequent and mild. There were hardly any influenza-like symptoms and the quality of life improved. In the group who volunteered to have their immunological state checked, significant boosting effects were found. It was concluded that there might be a good indication for using suitable herbal preparations as a means of preventing influenza-like infection. The mode of preventive effect could be treatment of the infection at its very early stage instead of producing a period of higher immunological ability, as in the case of vaccination. PMID- 16047554 TI - Anti-hypertensive effect of chunghyul-dan (qingxue-dan) on stroke patients with essential hypertension. AB - Hypertension is one of the modifiable risk factors for stroke. Lowering blood pressure is helpful for primary and secondary prevention of stroke. This study is aimed to assess the efficacy of Chunghyul-dan on stroke patients with stage 1 hypertension using 24 hours ambulatory blood pressure monitoring (24ABPM). Forty hospitalized stroke patients with stage 1 hypertension were included in the study and they were randomly assigned into two groups: group A was treated with Chunghyul-dan 1200 mg once a day for 2 weeks, while group B was not. Twelve subjects were dropped out because of unexpected early discharge or data errors, thus the remaining 28 subjects were included in the final analysis (15 in group A and 13 in group B). Blood pressure was monitored every 30 minutes for 24 hours at baseline and 2 weeks after medication. Blood pressure, pulse rate, trough/peak ratio (TPR) [the value calculated by dividing the blood pressure change at trough (22 to 24 hours after drug intake) by the change at peak (2 adjacent hours with a maximal blood pressure reduction between the second and eighth hour after drug intake)] and smoothness index (SI) (the value calculated as the ratio between the average of the 24 hours, treatment-induced blood pressure changes and its standard deviation) were compared to assess the efficacy of Chunghyul-dan. To assess the safety of Chunghyul-dan, any adverse effects during medication period were monitored. There was no significant difference in the baseline assessment between the two groups. Systolic blood pressure was lower in group A than in group B (141.37+/-8.96 mmHg versus 132.28+/-9.46 mmHg, P = 0.03), while diastolic blood pressure and pulse rate had no significant difference between the two groups. Systolic TPR and SI was 0.87 and 1.04 in group A, respectively. This suggests that Chunghyul-dan have anti-hypertensive effect on stroke patients with stage 1 hypertension. PMID- 16047555 TI - Traditional Chinese medicine based subgrouping of irritable bowel syndrome patients. AB - Partly from lack of effective conventional therapeutics, patients with irritable bowel syndrome (IBS) turn to complementary and alternative approaches, including Traditional Chinese Medicine (TCM). Key to TCM's approach to IBS is individualized therapies targeted at subgroups. Subgroups represent distinct patterns of dysregulation (e.g. "excess" or "deficiency") identified by both intestinal and extra-intestinal symptoms. Our objective was to identify operational criteria supporting the existence of TCM-based subgroups in IBS and to assess reliability and validity of these criteria. Using TCM principles, items were selected on face validity from conventional questionnaires. TCM practitioners evaluated items for content and face validity. Symptom items and a set of patient cases with item responses were validated by examining patient's pattern of response to items and assessing the consistency with which practitioners diagnosed patients on the spectrum of an "excess" or "deficiency" syndrome. Standard correlation analysis revealed 33 intestinal and extra intestinal symptom items. There was high degree of practitioner agreement in assessing individual items to particular patterns. External validation by practitioners of cases showed high internal consistency among practitioners (Cronbach's alpha coefficients of 0.91 and 0.87 for excess and deficiency, respectively) and high correlation of average practitioner rating to original questionnaire generated scores (Pearson correlation coefficients of 0.94 and 0.92 for excess and deficiency, respectively). This pilot study provides preliminary support for a methodology to identify novel subgroups of IBS patients related to the TCM classification, which may differ in underlying pathophysiology and treatment responses. PMID- 16047556 TI - Immunomodulatory activities of Yunzhi and Danshen in post-treatment breast cancer patients. AB - Breast cancer is the most common cancer among women worldwide. Discomfort and fatigue are usually arisen from anticancer therapy such as surgery, radiotherapy, chemotherapy, hormonal therapy, or combination therapy, because of the suppressed immunological functions. Yunzhi (Coriolus versicolor) can modulate various immunological functions in vitro, in vivo, and in human clinical trials. Danshen (Salvia miltiorrhiza) has been shown to benefit the circulatory system by its vasodilating and anti-dementia activity. The purpose of this clinical trial was to evaluate the immunomodulatory effects of Yunzhi-Danshen capsules in post treatment breast cancer patients. Eighty-two patients with breast cancer were recruited to take Yunzhi [50 mg/kg body weight, 100% polysaccharopeptide (PSP)] and Danshen (20 mg/kg body weight) capsules every day for a total of 6 months. EDTA blood samples were collected every 2 months for the investigation of immunological functions. Flow cytometry was used to assess the percentages and absolute counts of human lymphocyte subsets in whole blood. Plasma level of soluble interleukin-2 receptor (sIL-2R) was measured by enzyme-linked immunosorbent assay (ELISA). Results showed that the absolute counts of T-helper lymphocytes (CD4+), the ratio of T-helper (CD4+)/T suppressor and cytotoxic lymphocytes (CD8+), and the percentage and the absolute counts of B-lymphocytes were significantly elevated in patients with breast cancer after taking Yunzhi Danshen capsules, while plasma slL-2R concentration was significantly decreased (all p < 0.05). Therefore, the regular oral consumption of Yunzhi-Danshen capsules could be beneficial for promoting immunological function in post treatment of breast cancer patients. PMID- 16047557 TI - Ginseng and diabetes. AB - Ginseng is a well-known medicinal plant used in traditional Oriental medicine. In recent decades, ginseng root has gained popularity as a dietary supplement in the United States. Ginseng has also been commonly used in Oriental medicine to treat diabetes-like conditions. The present review discusses the research on the anti diabetic effects of ginseng and the possible mechanisms of its anti-diabetic actions. PMID- 16047558 TI - Caesalpinia sappan induces cell death by increasing the expression of p53 and p21WAF1/CIP1 in head and neck cancer cells. AB - Caesalpinia sappan L. (C. sappan) has been used in Oriental medicine as an antitumor agent. The present study shows the effects of the chloroform extract of C. sappan on cell death in head and neck cancer cell lines. The viability of HNSCC4 and HNSCC31 cells (head and neck cancer cell lines) was noticeably decreased compared to that of HaCaT cells (control group) in the presence of chloroform extract. No significant difference was observed in the viability of HNSCC4 and HNSCC31 cells when compared with HaCaT cells in the presence of n butanol, methanol, and water extracts. Exposure to the chloroform extract of C. sappan resulted in an increase in the Sub-G1 phase of the cell cycle and condensation and shrinkage of nuclei in the HNSCC4 and HNSCC31 cells. The levels of p53 and p21WAF1/CIP1 were also increased in the HNSCC4 and HNSCC31 cells. The results suggest that the chloroform extract of C. sappan may increase cell death in the HNSCC4 and HNSCC31 cells, which is linked to increased cellular levels of p53 and p21WAF1/CIP1. PMID- 16047559 TI - Anti-inflammatory activity of herbal medicines: inhibition of nitric oxide production and tumor necrosis factor-alpha secretion in an activated macrophage like cell line. AB - Houttuynia cordata Thunb. (HC), Glycyrrhiza uralensis Fischer (GU), Forsythia suspense (Thunb.) Vahl (FS), and Lonicera japonica Thunb. (LJ) are Chinese herbs known to possess anti-inflammatory properties. The effects of aqueous extracts of these herbs on the production of the pro-inflammatory mediators, nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) were examined in an activated macrophage-like cell line, RAW 264.7 cells. Aqueous extracts from FS at 0.0625 2.0 mg/ml inhibited in vitro production of NO and secretion of TNF-alpha in a dose-dependent manner. FS at 1.0-2.0 mg/ml and 0.125-2.0 mg/ml significantly inhibited NO production and TNF-alpha, respectively. An extract of LJ demonstrated potent inhibition of both NO production and TNF-alpha secretion in a dose-dependent manner. An aqueous extract from HC inhibited NO production in a dose-dependent manner, but minimally (approximately 30%) inhibited TNF-a secretion at 0.0625 and 0.125 mg/ml. In contrast, an aqueous extract of GU had a minimal effect on both the production of NO and the secretion of TNF-alpha. Viability of cells at all concentrations studied was unaffected as determined by MTT cytotoxicity assay and trypan blue dye exclusion. These results suggest that aqueous extracts from FS, LJ and HC have anti-inflammatory actions as measured by inhibition of NO production and/or TNF-alpha secretion. PMID- 16047560 TI - Microglia, apoptosis and interleukin-1beta expression in the effect of sophora japonica l. on cerebral infarct induced by ischemia-reperfusion in rats. AB - Sophora Japonica L. (SJ) is a traditional Chinese herb used to cool blood, stop bleeding and to treat hemorrhoids with bleeding. Although several recent studies found that both SJ and Ginkgo biloba have the same components of quercetin and rutin, only Ginkgo biloba has been widely used to treat cerebrovascular disorders and dementia in humans. This study investigated the effect of SJ on cerebral infarct in rats. A total of 66 Sprague-Dawley (SD) rats were studied. Focal cerebral infarct was established by occluding the bilateral common carotid arteries and the right middle cerebral artery for 90 minutes. After 24 hours of reperfusion, the neurological status was evaluated. The rats were then killed, and brain tissue was stained with 2,3,5-triphenyl-tetrazolium chloride. The grading scale of neurological deficit and the ratio of cerebral infarction area were used as an index to evaluate the effect of SJ on cerebral infarct. In addition, the number of ED1 and interleukin-1beta immunostaining positive cells, and apoptotic cells were measured in the cerebral infarction zone. The results indicated that pre-treatment with 100 or 200 mg/kg SJ and post-treatment with 200 mg/kg SJ significantly reduced the grade of neurological deficit and the ratio of cerebral infarction area. In addition, pre-treatment with 200 mg/kg SJ also significantly reduced ED1 and interleukin-1beta immunostaining positive cells, and apoptotic cells in ischemia-reperfusion cerebral infarct rats. This study demonstrated that SJ could reduce the cerebral infarction area and neurological deficit induced by ischemia-reperfusion in rats, suggesting its potential as a treatment for cerebral infarct in humans. This effect of SJ involves its suppressive action of microglia, interleukin-1beta and apoptosis. PMID- 16047561 TI - Inhibitory effects of bezoar bovis on intimal formation and vascular smooth muscle cell proliferation in rat. AB - Intimal formation of animal carotid arteries induced by balloon endothelial denudation has been considered to be an "accelerated atherosclerosis" model and used in primary screening methods to evaluate natural drugs and chemical candidates. The aim of the present study was to examine whether intimal formation is prevented by Bezoar Bovis (dried cattle gallbladder stones: Niuhuang in Chinese and Go-o in Japanese), which has been used to prevent heart palpitation in patients with hypertension. The intimal-to-medial area ratio in rat carotid arteries 7 days after balloon endothelial denudation was significantly reduced by oral administration of Bezoar Bovis. Bezoar Bovis also suppressed vascular smooth muscle cells (VSMCs) proliferation, which is thought to play important roles in the intimal formation after endothelial damage and also atherosclerosis resulting from long-term inappropriate lifestyle. The present findings suggest that Bezoar Bovis may be useful for preventing atherosclerosis and for protection against restenosis after percutaneous coronary intervention, for which effective reduction method is not currently available. PMID- 16047562 TI - Effect of curcuma herbs on vasomotion and hemorheology in spontaneously hypertensive rat. AB - Curcuma herbs have a vasodilator effect. The effects of C. longa, which induces only endothelium-independent vasodilatation, and C. zedoaria, which induces both endothelium-dependent and -independent vasodilatation, were studied on vasomotion and hemorheology in spontaneously hypertensive rats. Spontaneously hypertensive eight-week-old male rats were assigned to five groups. For 12 weeks, the control group received standard chow. The 3%CL (C. longa) group received standard chow containing 3% (wt/wt) C. longa. The 1%CZ and 3%CZ (C. zedoaria) groups received standard chow containing 1% and 3% (wt/wt) C. zedoaria, respectively. The captoril group received standard chow and 100 mg/kg/day of captoril in drinking water. Blood pressure, vasomotion, hemorheology, etc. were examined. Systolic blood pressure of the 3%CZ and captoril groups decreased significantly as compared to the control group. Acetylcholine-induced endothelium-dependent relaxations of the 3%CZ and captoril groups were increased to a greater degree, significantly, than the control group. When testing xanthine oxidase-induced contraction, the 3%CZ group was significantly decreased as compared to the control group. Low shear stress of whole blood viscosity showed the 3%CL and 3%CZ groups to be decreased significantly compared to the control group. Thus, Curcuma herbs have hypotensive and protective effect on the endothelium in spontaneously hypertensive rats. Especially, C. zedoaria is more effective than C. longa, and its mechanism is thought to be related to a radical scavenging effect and improvement of hemorheology. PMID- 16047563 TI - Genomic expression for rat model of damp obstruction in Chinese medicine: application of microarray technology. AB - Damp obstruction refers to the stagnation of vital energy (qi) caused by dampness resulting in dysfunction of body and limbs movement, as well as impairment of spleen and stomach digestive function. Damp obstruction is the dampness-induced imbalance of five elements; thus it serves as an ideal model for genomic study using cDNA microarray. We have performed microarray analyses to major organs of damp-obstructed rats. Cluster analysis for the expression profiles of major organs indicated that spleen, stomach, and kidney respond to dampness differently from heart, liver, lung, and brain. Gene expression profile specific to each element or group of elements was also identified. Our results are consistent with the philosophy of Chinese medicine that the five elements, metal (lung), wood (liver), water (kidney), fire (heart), and earth (spleen and stomach) coordinate by subjugation or restriction to maintain a healthy, physiological state. This is the first time that a powerful genomic tool was applied to probe the ancient theory of Chinese medicine. PMID- 16047564 TI - Post-ischemic treatment with toki-shakuyaku-san (tang-gui-shao-yao-san) prevents the impairment of spatial memory induced by repeated cerebral ischemia in rats. AB - Previously we have reported that Toki-shakuyaku-san (TSS) ameliorated the impairment of spatial memory induced by single cerebral ischemia (1 x 10 minutes) and scopolamine, a muscarinic receptor antagonist. In this experiment, we studied the effect of TSS on repeated cerebral ischemia (2 x 10 minutes, 1-hour interval) induced impairment of spatial memory and neuronal injury in rats. The 8-day post ischemic treatment with TSS (30-300 mg/kg) was administered p.o. once per day. TSS dose-dependently prevented the impairment of spatial memory, neuronal death and TUNEL positive cells induced by repeated cerebral ischemia. In order to determine the mechanism of TSS, we also studied the effect of TSS on GluR2 mRNA, one of the glutamate alpha-amino-3-hydroxy-5-methyl-4-isoxazole (AMPA) receptor subunits. Repeated cerebral ischemia significantly decreased GluR2 flop mRNA at 1 and 3 days after the occlusion. TSS (300 mg/kg) significantly suppressed the decrease in GluR2 flop at 3 days after repeated cerebral ischemia. These results suggested that the TSS has neuroprotective action which may be indirectly mediated by the AMPA receptor, and TSS may be beneficial for the treatment of cerebrovascular dementia. PMID- 16047565 TI - The nephroprotective effects of the herbal medicine preparation, WH30+, on the chemical-induced acute and chronic renal failure in rats. AB - In this study, we evaluated the renal protective effects of a Chinese herbal preparation WH30+ in male Wistar rats with glycerol-induced acute renal failure and adenine-induced chronic renal failure. WH30+ is a Chinese herb preparation composed of Rheum Palmatum, Salvia Miltiorrhiza, Cordyceps Sinensis, Leonurus Sibiricus, Epihedium Macranthum, Radix Astragali, and Radix Codonopsis Pilosulae, which has been used to treat kidney deficiency in human. An acute renal failure and chronic renal failure rat model were introduced by glycerol injection (i.m.) and fed with adenine-excessive diet, respectively. WH30+ was administered to rats at the dose of 50 mg/kg/day from 10 days before the diseases were induced until the rats were sacrificed. A reduction in body weight (p < 0.01) was observed in rats with chronic renal failure, but there was no difference between treatment groups. However, the body weight of rats with acute renal failure without treatment was significantly lower than those treated with WH30+ (p < 0.05). Overall, serum creatinine and urea nitrogen were elevated significantly (p < 0.01) in renal failure rats compared to control. Treatment with WH30+ improved both serum creatinine and urea nitrogen slightly in both models. The WH30+ treated rats with acute renal failure had significantly (p < 0.05) greater creatinine clearance than those without treatment. The results of the study show that WH30+ is more effective in the prevention of acute renal failure than chronic renal failure. PMID- 16047566 TI - Novel approach of molecular genetic understanding of iridology: relationship between iris constitution and angiotensin converting enzyme gene polymorphism. AB - Iridology is the study of the iris of the eye to detect the conditions of the body and its organs, genetic strengths and weaknesses, etc. Although iridology is not widely used as a scientific tool for healthcare professionals to get to the source of people's health conditions, it has been used as a supplementary source to help the diagnosis of medical conditions by noting irregularities of the pigmentation in the iris among some Korean Oriental medical doctors. Angiotensin converting enzyme (ACE) gene polymorphism is one of the most well studied genetic markers of vascular disease. We investigated the relationship between iridological constitution and ACE polymorphism in hypertensives. We classified 87 hypertensives and 79 controls according to iris constitution and determined the ACE genotype of each individual. DD genotype was more prevalent in patients with a neurogenic constitution than in controls. This finding supports the hypothesis that D allele is a candidate gene for hypertension and demonstrates the association among ACE genotype, Korean hypertensives and iris constitution. PMID- 16047567 TI - Is there any energy transfer during acupuncture? AB - Acupuncture therapy is based on the principle of restoring equilibrium of the body's energy state by regulating the flow of Qi in the corresponding meridians and acupoints. The purpose of this study was to determine whether bio-energy transfer occurs during acupuncture of the meridians. We treated 20 normal healthy subjects with acupuncture and measured changes in the direct current (DC) potential between the stomach meridian points ST-39 and ST-37 in response to invasive insertion of a needle at ST-36. Using a crossover study design, the practitioner performed the procedure on each subject once with bare (uncovered) hands (ABH) and once wearing surgical gloves (ASH) to shield the electric energy transfer. ABH produced a response pattern to each stimulation, whereas ASH produced a response pattern only to the first step. ABH elicited a significantly higher response potential than did ASH (p < 0.001). These results suggest that there may be bio-energy (Qi) transfer during acupuncture along the meridians. This transferred bio-energy (Qi) seems to act as an electromotive force, which is purported to remove the stagnation or blockage of energy and restore an equilibrium state. PMID- 16047568 TI - Evolutionary development of acupuncture theory. PMID- 16047569 TI - Charting a course to competency. PMID- 16047570 TI - Competencies for the physician assistant profession. PMID- 16047571 TI - The second time around. PMID- 16047572 TI - Enhancing contraception: a comprehensive review. AB - This review covers familiar and emerging contraceptive methods, comparing the risks and benefits of each. These basics will help you connect patients with the most acceptable form of contraception for them. PMID- 16047573 TI - Injury-related causes of acute knee pain. AB - This detailed overview covers the anatomy, function, and examination of the knee and describes the phases of treatment for acute knee pain secondary to injury. PMID- 16047574 TI - Case report: classic Kaposi's sarcoma. AB - Classic KS usually appears between ages 50 and 70 years and has a relatively benign, indolent course for 15 years or longer. Treatment usually controls the disease quite well. PMID- 16047575 TI - Leishmaniasis: early diagnosis is key. AB - A patient presents with nasal congestion, a history of nosebleed, and painful lesions on his body and in his mouth. What questions should you ask to ensure that you make an accurate diagnosis? PMID- 16047576 TI - An "infection" that antibiotics won't cure. PMID- 16047577 TI - Case of the month. Histiocytic necrotizing lymphadenitis. PMID- 16047578 TI - Lower embryonic loss rates among twin gestations following assisted reproduction. AB - PURPOSE: To determine whether maternal age and number of transferred embryos influence early pregnancy losses in twin pregnancies compared to singletons following IVF/ICSI. METHODS: We compared the pregnancy loss rates in singleton (n = 549) and twin (n = 252) gestations, stratified by maternal age (< or = 35 and > 35 years) and the number of transferred embryos (1-3 and 4-9). RESULTS: Loss rates of singleton pregnancies were significantly higher than that in twins (OR 3.0, 95% CI 1.9, 4.9), especially among singletons conceived after transfer of 4 9 embryos (OR 5.0, 95% CI 2.2, 11.9). Younger mothers of twins had lower loss rates (OR 0.3, 95% CI 0.1, 0.9). CONCLUSION: Twins have a significantly reduced spontaneous miscarriage rate compared with singletons following IVF/ICSI. Higher implantation rates per cycle (i.e., development of twins rather than one live embryo) may represent a better capacity of the uterus for early embryonic development. PMID- 16047579 TI - Clinical experiences of ICSI-ET thawing cycles with embryos cryopreserved at different developmental stages. AB - PURPOSE: The aim of our study was to analyze factors including survival, implantation and pregnancy rate, patients' age and BMI, abortions and extra uterine pregnancies that might influence the outcome of ICSI-ET thawing cycles. METHODS: A total of 147 cycles with embryos cryopreserved at different developmental stages were retrospectively evaluated. RESULTS: No difference was found in the survival, implantation and pregnancy rates of embryos cryopreserved on Day 2-3 and 5. However, in the pregnant group significantly higher implantation rate was observed with Day 5 blastocysts then with Day 2 or 3 early embryos. We found no difference in the number of abortions and extra uterine pregnancies between fresh and frozen ICSI-ET cycles. Higher BMI was found in the pregnant than in the nonpregnant group. However, the age of patient had no effect on the results. CONCLUSIONS: Developmental stage of embryo and patients' BMI influences the success of ICSI-ET thawing cycles. PMID- 16047580 TI - The influences of weather on patients with different ovarian responses in the treatment of assisted reproductive technology. AB - PURPOSE: The purpose of this study was to evaluate the influences of tropical weather on patients with different ovarian responses in the treatment of assisted reproductive technology. METHODS: Six-hundred fourty-seven women underwent their first treatment cycles were retrospectively analyzed. Patients received embryo transfer either 3 days or 5 days after oocyte retrieval, depending on the number and quality of embryos on day-2. RESULTS: Significant correlations were demonstrated in the top quality embryo rates of day-3 and day-5 embryo transfers with temperature, humidity, and atmosphere pressure. The cumulative light hours negatively correlated with the implantation and pregnancy rates of day-3 embryo transfer (-.282 and -.282, respectively), while they positively correlated with those of day-5 embryo transfer (.225 and .224, respectively). CONCLUSIONS: These results clearly suggest that weather may exert influences on the outcome of assisted reproductive technology. Patients with different ovarian responses or blastocyst culture and transfer may modify weather influences. PMID- 16047581 TI - Defining women who are prone to have twins in in vitro fertilization--a necessary step towards single embryo transfer. AB - PURPOSE: The aim of the present study was to identify subsets of women undergoing two-embryo transfer in in vitro fertilization (IVF) who are prone to give birth to twins. METHODS: During 1990 to 2002 4580 day 2 or day 3 two-embryo transfers were conducted and these constituted the study population. RESULTS: By selecting combinations of factors, some subsets of patients where single-embryo transfer might be conducted would reduce the estimated twin rate by 60%, but requiring single-embryo transfer in only approximately one third of the patients. Examples of such selected groups were patients less than 33 years of age with two top quality embryos, patients less than 39 years of age with two top quality embryos and conducting their first IVF cycle, and patients less than 39 years of age with two top quality embryos and two optimal cleavage cells. CONCLUSION: By combining factors, subsets of patients with both a high birth and twin rate that could be recommended single embryo transfer were identified. PMID- 16047582 TI - Insulin-like growth factor-1 and insulin-like growth factor binding protein-1 and 3 in the follicular fluid of infertile patients submitted to in vitro fertilization. AB - PURPOSE: In the present article we propose to evaluate IGF-1, IGFBP-1 and 3 in the follicular fluid of infertile patients submitted to in vitro fertilization. METHODS: We performed a case-control study with 53 infertile patients submitted to the first in vitro fertilization attempt. We compared their follicular fluid concentration of IGF-1, IGFBP-1 and IGFBP-3 between the patients who became pregnant (n = 11) versus those nonpregnant (n = 42). RESULTS: The clinical characteristics of patients from the two groups were similar in terms of age and body mass index. Data related to the analysis of ovulation induction was not different regarding length of induction in days, number of retrieved oocytes, fertilization rate, and number of transferred embryos. Furthermore, the number of FSH units required for ovarian induction was also similar between the studied groups. IGF-1 and IGFBP-1 were not significantly different between the groups (p > 0.05). However, those patients that became pregnant presented a lower follicular fluid concentration of IGFBP-3, 2237.10 +/- 582.73 pg/ml and 2657.64 +/- 584.15 ng/ml, respectively (p = 0.038). CONCLUSIONS: We demonstrated an association of a lower follicular fluid IGFBP-3 in individuals that became pregnant compared to subjects that did not after in vitro fertilization. PMID- 16047583 TI - Luteal phase serum cell-free DNA as a marker of failed pregnancy after assisted reproductive technology. AB - PURPOSE: DNA-damaging factors have been reported in patients that failed to achieve pregnancy after assisted reproductive technologies (ART). The hypothesis was that increased circulating cell-free DNA released by damaged cells could predict unfavorable conditions leading to failed ART treatment. The objective was to compare the relative concentrations of cell-free DNA in the luteal phase sera of nonpregnant versus pregnant patients. METHODS: Frozen-thawed sera (30 IVF cases) were obtained 1 week after embryo transfer. There were 16 pregnant and 14 nonpregnant cases and controls consisting of male sera (n = 8 cases). Modified isocratic capillary electrophoresis was performed and the images analyzed for cell-free DNA. RESULTS: Circulating cell-free DNA were identified in the sera of all patients. The serum concentrations of high (12 kb) and low (1 kb) molecular weight cell-free DNA were similar for both nonpregnant and pregnant patients. Male control sera had higher cell-free DNA concentrations compared with females. Evaluation of sera from a control case showed no fluctuations in cell-free DNA concentrations throughout specific days of the menstrual cycle. CONCLUSIONS: The results do not support the use of the luteal phase cell-free DNA concentration as a marker for failed pregnancies. The equal concentrations of high and low molecular weight cell-free DNA and ladder band-like gel patterns suggested cell apoptosis as the source of DNA. PMID- 16047584 TI - Developing techniques for determining sperm morphology in today's andrology laboratory. AB - PURPOSE: To investigate three areas: the staining of spermatozoa; the Computer Assisted Semen Analysis (CASA); and the variability of technicians. METHODS: Staining experiment: sperm from 15 beef bulls were randomized to one of three staining protocols. CASA experiment: slides were evaluated using the Integrated Visual Optical System and compared to technician results. Variability of technicians: five laboratorians analyzed the same set of 20 slides six different times. RESULTS: Staining experiment: the size of the sperm increased in proportion to increased time and heat associated with each successive protocol. CASA experiment: coefficient of variation ranged from 18.3 to 101.7% (12 slides). Variability of technicians: the mean sperm morphology results ranged from 7.3 to 15% normal forms. CONCLUSIONS: Until laboratories adhere to the universal standard set by the World Health Organization to evaluate sperm morphology, a laboratory must rely on its own quality control to insure repeatable results. PMID- 16047585 TI - Is it worthwhile to operate on subclinical right varicocele in patients with grade II-III varicocele in the left testicle? AB - PURPOSE: To determine whether repair of subclinical varicoceles in the right testicle results in significant seminal improvement in patients with clinical left varicocele. METHODS: Patients were divided into two groups: Group I (unilateral varicocelectomy) and Group II (bilateral varicocelectomy-subclinical left varicocele). The mean sperm concentration before treatment was higher in Group I (21.01 +/- 19.1) compared to Group II (5.7 +/- 10.7) (p = 0.04). RESULTS: An increase in volume was detected in the left testicle of patients in Group I (17 +/- 7.9 vs. 22.81 +/- 8.2; p = 0.04) and in the right testicle of patients in Group II (18.4 +/- 6.2 vs. 22.3 +/- 6.5; p = 0.04). Although the mean postoperative sperm concentration in Group I increased slightly (25.7 +/- 22.8), the mean sperm concentration in Group II increased significantly (30.32 +/- 9.8; p = 0.03). Pregnancy rate was higher in Group II (66.7%) compared to Group I (33.3%). CONCLUSIONS: Even a small, subclinical unrepaired varicocele continues to have a detrimental effect on bilateral testis function in a patient with grade II-III left varicocele. PMID- 16047586 TI - Slugs: what do you do with a lazy crew? PMID- 16047587 TI - Beyond competent. PMID- 16047588 TI - Triage at a major incident. PMID- 16047589 TI - The boil bites back. PMID- 16047590 TI - "Just the flu". PMID- 16047591 TI - Get the score so "everybody" wins! PMID- 16047592 TI - Specialized vehicles in EMS: MCI vehicles. PMID- 16047593 TI - Ambulance crashes: fatality factors for EMS workers. PMID- 16047594 TI - Renal failure and dialysis patients: What the EMS provider should know. PMID- 16047595 TI - Immunity statutes: how State laws protect EMS providers. PMID- 16047596 TI - The Alzheimer's challenge. AB - We underestimate our ability to communicate with and treat patients with Alzheimer's. Knowledge about the disease's origin, progression and symptomology can help EMS providers effectively communicate and treat patients with dementia. PMID- 16047597 TI - Practical techniques for effective volunteer recruitment. PMID- 16047598 TI - Handling difficult employees. PMID- 16047599 TI - Delivery of quality services. PMID- 16047600 TI - Practical leadership. PMID- 16047601 TI - Station on the hill. PMID- 16047602 TI - Facilities find subsites provide alternative home for focused content. PMID- 16047603 TI - Portal development. PMID- 16047604 TI - How one healthcare organization created an uncluttered home page. PMID- 16047605 TI - Pfizer sponsors study on usability for low-literacy users. PMID- 16047606 TI - Unsignaled delay of reinforcement, relative time, and resistance to change. AB - Two experiments with pigeons examined the effects of unsignaled, nonresetting delays of reinforcement on responding maintained by different reinforcement rates. In Experiment 1, 3-s unsignaled delays were introduced into each component of a multiple variable-interval (VI) 15-s VI 90-s VI 540-s schedule. When considered as a proportion of the preceding immediate reinforcement baseline, responding was decreased similarly for the three multiple-schedule components in both the first six and last six sessions of exposure to the delay. In addition, the relation between response rates and reinforcement rates was altered such that both parameters of the single-response version of the matching law (i.e., k and Re) were decreased. Experiment 2 examined the effects of unsignaled delays ranging from 0.5 s to 8.0 s on responding maintained by a multiple VI 20-s VI 120 s schedule of reinforcement. Response rates in both components increased with brief unsignaled delays and decreased with longer delays. As in Experiment 1, response rates as a proportion of baseline were affected similarly for the two components in both the first six and last six sessions of exposure to the delay. Unlike delays imposed between two stimulus events, the effects of delays between responses and reinforcers do not appear to be attenuated when the average time between reinforcers is longer. In addition, the disruptions produced by unsignaled delays appear to be inconsistent with the general finding that responding maintained by higher rates of reinforcement is less resistant to change. PMID- 16047607 TI - Effects of reinforcement history on response rate and response pattern in periodic reinforcement. AB - Several researchers have suggested that conditioning history may have long-term effects on fixed-interval performances of rats. To test this idea and to identify possible factors involved in temporal control development, groups of rats initially were exposed to different reinforcement schedules: continuous, fixed time, and random-interval. Afterwards, half of the rats in each group were studied on a fixed-interval 30-s schedule of reinforcement and the other half on a fixed-interval 90-s schedule of reinforcement. No evidence of long-term effects attributable to conditioning history on either response output or response patterning was found; history effects were transitory. Different tendencies in trajectory across sessions were observed for measures of early and late responding within the interreinforcer interval, suggesting that temporal control is the result of two separate processes: one involved in response output and the other in time allocation of responding and not responding. PMID- 16047608 TI - The effects of interval duration on temporal tracking and alternation learning. AB - On cyclic-interval reinforcement schedules, animals typically show a postreinforcement pause that is a function of the immediately preceding time interval (temporal tracking). Animals, however, do not track single-alternation schedules-when two different intervals are presented in strict alternation on successive trials. In this experiment, pigeons were first trained with a cyclic schedule consisting of alternating blocks of 12 short intervals (5 s or 30 s) and 12 long intervals (180 s), followed by three different single-alternation interval schedules: (a) 30 s and 180 s, (b) 5 s and 180 s, and (c) 5 s and 30 s. Pigeons tracked both schedules with alternating blocks of 12 intervals. With the single-alternation schedules, when the short interval duration was 5 s, regardless of the duration of the longer interval, pigeons learned the alternation pattern, and their pause anticipated the upcoming interval. When the shorter interval was 30 s, even when the ratio of short to long intervals was kept at 6:1, pigeons did not initially show anticipatory pausing-a violation of the principle of timescale invariance. PMID- 16047609 TI - Effects of methylphenidate and morphine on delay-discount functions obtained within sessions. AB - Four rats responded under a "self-control" procedure designed to obtain delay discount functions within sessions. Each session consisted of seven blocks, with seven trials within each block. Each block consisted of two initial forced-choice trials followed by five free-choice trials. On choice trials, the rats could press either of two retractable levers. A press on one lever was followed by presentation of a smaller reinforcer (a single dipper presentation of a sucrose solution); a press on the other lever was followed by presentation of a larger reinforcer (four consecutive dipper presentations). The delay associated with the smaller reinforcer always was 0 s, whereas the signaled delay associated with the larger reinforcer increased across blocks (from 0 to 50 s). Under these conditions, the percentage of choices of the larger reinforcer decreased across blocks, and relatively reliable delay-discount functions were obtained within sessions. Doses of methylphenidate (1.0 to 17.0 mg/kg) and morphine (0.3 to 17.0 mg/kg) were then administered prior to selected sessions. Typically, intermediate doses of methylphenidate shifted the discount functions to the right (increased choices of the larger reinforcer). For 2 of the rats, this effect was pronounced; for the other 2 rats, this effect occurred after the range of delays for the larger reinforcer was decreased (0 to 20 s). On the other hand, in most cases morphine produced a slight leftward shift in the discount function (decreased choices of the larger reinforcer). The present procedure appears to be a useful and efficient method to characterize drug effects on an entire delay-discount function. As with many procedures used to study self-control choices, however, sources of control other than reinforcement delay and amount may have been operating in the present study, and these sources must be considered when interpreting drug effects. PMID- 16047610 TI - Learning from leaders. PMID- 16047613 TI - Legal. Sharing the risk. AB - Negotiating IT contracts isn't for the faint of heart. Increasingly, hospitals are demanding that vendors share some of the financial and legal risk in case of product failure or an error. PMID- 16047614 TI - Coding & billing. Will codes turn 10? AB - All of health care continues to hold its collective breath waiting for the Health and Human Services Department to give its final blessing to a new and improved coding system, ICD-10. But that wait could go on for a few more years. PMID- 16047615 TI - Technology. Behind the hype. AB - Having proven successful in retail, military and other industries, radio frequency identification is making its way into health care. But can the technology live up to the bold claims? PMID- 16047616 TI - Patient care. Getting consent. PMID- 16047617 TI - Save lives now. Patient care. Transitioning care. AB - H&HN's ongoing series on ways that hospitals can reduce medical errors takes a look at how clinicians can better talk with one another as patients move from one setting to another. PMID- 16047618 TI - Data page. Eight financial measures every hospital executive should know. PMID- 16047619 TI - Capital catch-22. There is no magic number to guide investment. PMID- 16047620 TI - Capital: beyond the tried and true. AB - The old standbys remain by far the financing vehicles of choice for hospitals. But other, less traditional methods are gaining a foothold as not-for-profit hospitals look elsewhere--usually the corporate world--for successful precedents that can help fund big projects without leaving a bad financial aftertaste. PMID- 16047621 TI - Medication safety issue brief. Eliminating dangerous abbreviations, acronyms and symbols. AB - The use of abbreviations, acronyms and symbols in prescribing and transcribing medication orders too often results in the misinterpretation of the order's intent. Busy health care practitioners often use these shortcuts to indicate drug names, dosages, the patient's condition and route of administration. The result can be omission errors, extra or improper doses, administering the wrong drug, or giving a drug in the wrong manner. Stopping the use of unapproved abbreviations, acronyms and symbols can go a long way toward preventing these errors, but that's proven difficult to accomplish. This briefing examines ways that hospitals can put an end to the practice. PMID- 16047622 TI - Taking control of supply spending. AB - Bringing common sense to bear on buying medical supplies requires standardizing purchases. Adding RNs to your materials management staff can ease physicians' concerns. PMID- 16047623 TI - Looking harder. Not-for-profit hospitals use Sarbanes-Oxley to strengthen their boards' financial accountability. AB - The Sarbanes-Oxley Act is intended to improve corporations' financial accountability, but boards of not-for-profit hospitals are taking their own cues from its provisions. PMID- 16047624 TI - Integrating strategic and financial planning. AB - Our guide through the major steps for integrating strategic and financial planning shows how to balance organizational priorities and the deployment of resources. PMID- 16047625 TI - Diversity and the bottom line. Interview by Richard Haugh. AB - Fred Hobby, new head of the Institute for Diversity in Health Management, explains why cultural diversity is an organizational imperative in today's heterogenetic landscape. PMID- 16047626 TI - Everybody Supports the EHR. Now what? PMID- 16047627 TI - Show me the seed money. PMID- 16047628 TI - NHII: Achievable goal or house of cards. PMID- 16047629 TI - Will CMS be a leader or a nudger? PMID- 16047630 TI - Regional strategies for health info exchange. PMID- 16047631 TI - Should the patient be at the controls? PMID- 16047632 TI - State-by-state analysis shows how lack of insurance affects working Americans. PMID- 16047633 TI - National coordination urged for collection, use of stem cells from umbilical cords. PMID- 16047634 TI - Tomorrow's nurse. We all have a stake in the redefinition of nursing's role and education requirements. PMID- 16047635 TI - Spring. PMID- 16047636 TI - Now's the time. PMID- 16047637 TI - The dental home: a partial solution to the access challenge. PMID- 16047638 TI - Costs of individuals with disabilities and the elderly in Pennsylvania. AB - In 2000, an estimated 4,420 children (33 per 1,000 births) were born with a disability in Pennsylvania. In addition, more than 1.8 million Medicare enrollees (age 65 and older) were residents of the state. Selected estimated costs were developed for these two populations at state and county levels. PMID- 16047639 TI - Effect of soy protein-rich diet on renal function in young adults with insulin dependent diabetes mellitus. AB - BACKGROUND: Diabetic nephropathy is the most frequent cause of end-stage renal disease in the Western world. Dietary intake, including protein amount and type, seems to affect the progression of renal disease. This pilot study tested the hypothesis that substituting soy protein for animal protein in the diets of diabetics would help correct glomerular hyperfiltration. METHODS: Twelve young adults (aged 29.9 +/- 2.4 years) with type 1 diabetes mellitus (duration of diabetes 15.1 +/- 2.3 years) and hyperfiltration (glomerular filtration rate, GFR > 120 ml/min/1.73 m2) completed a crossover, dietary intervention trial. After a four-week assessment of baseline characteristics and dietary habits, subjects were assigned to either a control or soy diet for eight weeks after which each subject was crossed over to the alternative diet for another eight-week period. RESULTS: Mean GFR was significantly reduced (p < 0.02) after eight weeks on the soy diet (143 +/- 7.4 ml/min/1.73 m2) compared with baseline (159 +/- 7.7 ml/min/ 1.73 m2) and control diets (161 +/- 10.0 ml/min/1.73 m2). Urinary excretion of the soy isoflavones was significantly higher (p < 0.01) at the end of the soy diet (genistein 1,014.6 +/- 274.1 nmol/h, daidzein 2,645.1 +/- 989.6 nmol/h) compared with baseline (genistein 53.7 +/- 31.1 nmol/h, daidzein 151.1 +/- 74.1 nmol/h) and control diets (genistein 41.1 +/- 13.3 nmol/h, daidzein 127.5 +/- 54.0 nmol/h). The soy diet significantly reduced total and LDL cholesterol by 7% and 9%, respectively. CONCLUSIONS: Implementation of a soy-based diet appears to reduce the GFR and total and LDL cholesterol of young adults with type 1 diabetes and glomerular hyperfiltration, thus affecting positively their clinical profile. PMID- 16047640 TI - Adiponectin level is reduced and inversely correlated with the degree of proteinuria in type 2 diabetic patients. AB - AIMS: Adiponectin seems to be an important modulator for metabolic and vascular diseases. We aimed to measure plasma adiponectin levels in type 2 diabetic patients and investigate any association with the severity of proteinuria. METHODS: 80 patients (mean age, 46.9 +/- 5.1 years; body mass index (BMI), 25.8 +/- 1.98 kg/m2) and 47 healthy volunteers (mean age, 46.1 +/- 5.5 years; BMI 26.74 +/- 2.23 kg/m2) were included. Plasma adiponectin concentration, insulin levels, homeostasis model assessment (HOMA) indices, calculated glomerular filtration rate (GFR), high sensitive C reactive protein (hsCRP) and biochemistry panel were determined in all subjects. The association between adiponectin concentration and proteinuria was evaluated. Additionally, the relationship between adiponectin and hsCRP and calculated GFR were also investigated. RESULTS: Adiponectin levels in patients were significantly lower than those of controls (n = 80; 8.76 +/- 4.50 microg/ml for patients, n = 47; 24.27 +/- 5.59 microg/ml for controls, p < 0.001). Plasma adiponectin levels in patients with proteinuria were significantly lower than those without proteinuria (n = 43; 6.81 +/- 2.82 microg/ml for proteinuria, n = 37; 11.98 +/- 3.32 microg/ml for no proteinuria, p < 0.001). There was a significant negative correlation between plasma adiponectin concentrations and the degree of proteinuria (r = -0.433, p < 0.001). There were also significant negative correlations between adiponectin concentrations and insulin levels as well as HOMA index in the patient group (r = -0.322, p = 0.004; r = -0.301, p = 0.032). Additionally there was a significant negative correlation between adiponectin and hsCRP levels in the patient group (r = -0.872, p < 0.001). CONCLUSION: The results show that adiponectin is lower in patients with type 2 diabetes and the levels are negatively correlated with the severity of proteinuria. PMID- 16047641 TI - Association between polymorphisms of the renin-angiotensin system and more severe histological forms of lupus nephritis. AB - AIMS: The pathogenesis of lupus nephritis (LN) has not been fully understood. The renin-angiotensin system (RAS) is implicated in various immunological and non immunological phenomena, and the polymorphism of the RAS genes has been associated with cardiovascular and renal disease onset and outcome. Therefore, we evaluated the possible association between the polymorphism of the renin angiotensin system genes and the development of the different types of histological lesions of lupus nephritis in Brazilian patients. METHODS: 72 LN patients and 65 healthy subjects (sex-and ethnic-matched) were enrolled and compared in this study. Following the extraction of genomic DNA from the leukocytes of the peripheral blood, the genotypes of the angiotensin converting enzyme (ACE I/D), of the angiotensinogen (AGT M235T) and of the angiotensin II type 1 receptor (AGTR1 A1166C) were determined by the polymerase chain reaction. The renal lesions of the patients with LN were classified by the histological findings according to the WHO criteria. In addition, the activity and chronicity indices were used to assess the severity of renal involvement. RESULTS: Among the 72 patients with LN, there were 17 class II, 8 class III, 40 class IV and 7 class V, according to the WHO criteria. Individuals with the III and IV classes of LN (WHO) showed a significantly increased DD genotype frequency of ACE I/D genes when compared to the control group (48% vs. 27.7%, chi2 = 4.885, df = 1, p = 0.0442). No difference was found in the distribution of the AGT M235T and AGTR1 A1166C genotype frequencies among the LN of the different histological classes (WHO) and healthy controls. There was no association between genetic polymorphism of ACE, AGT M235T and AGTR1 A1166C and susceptibility to lupus nephritis, nor histological activity and chronicity indices in renal biopsy among the patients studied. CONCLUSIONS: This study suggests that the DD genotype of the ACE may be associated with the development of the more severe histological forms of lupus nephritis. PMID- 16047642 TI - Effects of long-term treatment with mizoribine in patients with proliferative lupus nephritis. AB - AIM: Mizoribine (MZR) is a purine antimetabolic immunosuppressant agent that has few little severe adverse events. We studied whether maintenance therapy with MZR and prednisolone (PSL) in severe proliferative lupus nephritis patients could improve immunity, reduce proteinuria, prevent renal relapse, and reduce steroid dose. METHOD: Long-term maintenance therapy with MZR and PSL was evaluated in ten patients with biopsy-proven proliferative lupus nephritis. Patients with severe lupus nephritis, who had proteinuria of 0.5 g or more even after treatments with plasma exchange and/or pulse methyl prednisolone, were recruited. MZR at an average dose of 140 +/- 10 (100 - 200) mg was administered two to three times/day in combination with PSL. The average period for the MZR maintenance therapy was 89.7 +/- 5.5 (70 - 126) months. Urine protein excretion, serum hemolytic complement activity (CH50), C3, serum creatinine, general and biochemical blood examinations, anti-ds-DNA antibody were collected at each monthly medical examination. RESULTS: All patients were females, mean age 43.0 +/- 3.3 years. A significant decrease in proteinuria was noted two years after the combination therapy (p = 0.0016). Five patients experienced lupus nephritis relapse. Patients who did not experience relapses had their MZR combination therapy initiated earlier (p = 0.037) when compared with the patients who experienced relapses. Serum creatinine levels remained unchanged in all patients throughout treatment and follow-up, even during renal relapses. Levels of C3 and CH50 normalized as proteinuria decreased. None of the patients developed serious side effects during MZR treatment. A significant steroid-sparing effect was observed three years after initiating MZR (p = 0.0025). CONCLUSION: From our long-term observation, maintenance therapy with low-dose PSL combined with MZR can eliminate proteinuria and have steroid-sparing effect. Early initiation of the therapy can protect against renal relapses among severe proliferative lupus nephritis patients without serious side effects. PMID- 16047643 TI - Treatment with low-dose angiotensin-converting enzyme inhibitor (ACEI) plus angiotensin II receptor blocker (ARB) in pediatric patients with IgA nephropathy. AB - AIM: IgA nephropathy associated with heavy proteinuria is considered a more progressive form of this disease. In this report, we describe the favorable clinical effect of combination therapy with low doses of an angiotensin converting enzyme inhibitor (ACEI) and angiotensin II receptor blocker (ARB) in the chronic stage of pediatric IgA nephropathy associated with heavy proteinuria. PATIENTS: We initially used ACEI for seven children with IgA nephropathy and heavy proteinuria who did not achieve remission with the routine treatment including steroids. RESULTS: With ACEI therapy alone, only three patients showed an antiproteinuric response. In one of the three patients, the proteinuria decreased by half, but was still over 1 g/day. In the other four patients, the proteinuria did not decrease. In these five patients, of whom one partial was a responder and four were non-responders for ACEI, ARB was added, and in marked contrast to ACEI therapy alone, the antiproteinuric effect was significantly augmented (p < 0.01). The antiproteinuric response induced by combination therapy was not accompanied by blood pressure changes. Urinary low-molecular protein and N-acetyl-beta-D-glucosaminidase (NAG) levels tended to decrease after both ACEI alone and combination therapy. CONCLUSION: These data indicate that inhibition therapy of the angiotensin system not only decreases proteinuria levels but also protects renal tubular cells. Moreover, there were no obvious side effects associated with this therapy during the follow-up period of our clinical trial. In conclusion, this report shows that the combination of low doses of ACEI and ARB might provide marked antiproteinuric and long-term renoprotective effects in pediatric IgA nephropathy, with this approach appearing to be both well-tolerated and safe. PMID- 16047644 TI - Serum cystatin C--a superior marker of rapidly reduced glomerular filtration after uninephrectomy in kidney donors compared to creatinine. AB - AIMS: Acute renal failure (ARF), defined by a rapid decrease of glomerular filtration rate (GFR), is associated with high mortality. Early and accurate detection of decreasing GFR is critical to prevent the progression of ARF and to potentially improve its outcome. Serum creatinine, the conventional GFR marker, has major limitations. We prospectively evaluated whether serum cystatin C detected a rapid GFR decrease earlier and more accurately than serum creatinine. METHODS: In ten patients undergoing nephrectomy for living related kidney transplantation, serum creatinine and cystatin C were determined daily. The decrease of GFR was quantitated preoperatively by creatinine clearance and MAG3 scintigraphy. The GFR decrease was defined by a 50-100% increase of cystatin C or creatinine from preoperative values. Ten patients without renal impairment served as controls. RESULTS: Initially, patients had a creatinine clearance of 105 +/- 14 ml/min/1.73 m2. Due to nephrectomy, patients lost 45 +/- 3% of their renal function. Serum cystatin C significantly increased already one, serum creatinine two days after nephrectomy. Cystatin C demonstrated an increase by 50-100% 1.4 +/ 0.9 days earlier than creatinine (p = 0.009). Serum cystatin C performed well detecting the GFR decrease with higher diagnostic values compared to creatinine. This was indicated by a sensitivity of 50, 70 and 80% of cystatin C to detect the GFR decrease on the three days following nephrectomy. CONCLUSIONS: Serum cystatin C detects rapid GFR decreases one to two days earlier than creatinine. Cystatin C is an early and accurate marker to detect rapid GFR decreases as in ARF. PMID- 16047645 TI - An ultrasonographic classification for diverse clinical symptoms of pediatric nutcracker phenomenon. AB - AIMS: The nutcracker phenomenon (NCP) is the significant compression of the left renal vein (LRV) by the aorta and superior mesenteric artery (SMA), and found in the patients with so-called idiopathic renal bleeding, orthostatic proteinuria and severe orthostatic intolerance. The purpose of this study is to investigate clinical implications among these disorders possibly related to the NCP. MATERIAL AND METHODS: We analyzed 93 pediatric patients (56 with idiopathic renal bleeding, 14 with massive orthostatic proteinuria and 23 with severe orthostatic intolerance), and the findings of 64 patients were compared on both digital subtraction angiography (DSA) and ultrasonography (US) to obtain a new US classification of the NCP with seven grades. RESULTS: US grades of the NCP were well-correlated with DSA findings (rs = 0.797, p < 0.0001). LRV stenosis was a typical finding in patients with idiopathic renal bleeding. LRV occlusion was observed in 70% for severe orthostatic intolerance, and in contrast in 18% and 14% for idiopathic renal bleeding and massive orthostatic proteinuria, respectively. Collateral veins on color Doppler US as well as a mirror image of the SMA in the aorta on conventional US were found as subsidiary signs of LRV occlusion. Extreme dilatation of the LRV was present in 44% for massive orthostatic proteinuria and in 7% for idiopathic renal bleeding (p < 0.0001). CONCLUSIONS: Anew US classification is useful for the diagnosis of the NCP in diverse clinical symptoms. PMID- 16047646 TI - Complete switch to darbepoetin in a hemodialysis unit. AB - AIMS: In March 2003, our hemodialysis unit switched all patients from subcutaneous (s.c.) rHuEPO to intravenous (i.v.) darbepoetin. The primary outcome was to assess the efficacy of i.v. darbepoetin to maintain target serum hemoglobin (Hb) compared to s.c. rHuEPO. Secondary outcomes were to evaluate the manufacturer's recommend guidelines for conversion of rHuEPO to darbepoetin, and to assess the cost implications of darbepoetin therapy. METHODS: This was an 18 month open-label observational study of 95 hemodialysis patients. At the time of the switch to darbepoetin (baseline), data were collected retrospectively for six months and prospectively for 12 months, at three-month intervals. The first six months of darbepoetin therapy was considered a dose titration phase, thus, data were analyzed comparing two six-month periods: (-) six months to baseline (rHuEPO phase) and (+) 6-12 months (darbepoetin phase). Doses were titrated to a target Hb of 120-135 g/l. RESULTS: There was no significant difference in Hb between phases at any time point. Mean Hb ranged from 119.6-121.5 g/l for rHuEPO and 121.9-123.4 g/l for darbepoetin. The median darbepoetin dose remained stable throughout the analysis at 30 microg/week while the median dose of rHuEPO rose from 8,000 U/week at minus six months to 9,000 U/week at baseline. Median 12 month cost savings associated with the administration of darbepoetin were estimated at 212,000 dollars. The recommended darbepoetin dose from the manufacturer's conversion table was deemed too low for baseline rHuEPO doses above 17,000 U/week. A more simplified dose conversion nomogram was created. CONCLUSION: Darbepoetin was able to maintain similar serum Hb levels compared to rHuEPO at a substantially reduced cost. PMID- 16047647 TI - Maxacalcitol therapy decreases circulating osteoprotegerin levels in dialysis patients with secondary hyperparathyroidism. AB - BACKGROUND: Osteoprotegerin is a natural glycoprotein which plays a critical role in osteoclast physiology. Elevated levels of circulating osteoprotegerin may account for the development of bone and mineral metabolic abnormalities in uremia. Little is known about the effects of vitamin D therapy on the circulating osteoprotegerin levels in dialysis patients. PATIENTS AND METHODS: Fifty chronic dialysis patients whose plasma intact PTH levels were greater than 300 pg/ml were analyzed for the study. Following a four-week washout time during which all vitamin D administration was halted, 10 microg of maxacalcitol was intravenously injected thrice a week. RESULTS: The circulating intact PTH, bone-specific alkaline phosphatase and intact osteocalcin levels were significantly lowered, while the serum calcium levels were elevated after the therapy. The osteoprotegerin levels significantly decreased after the therapy (p < 0.0001). CONCLUSION: Maxacalcitol therapy reduced the circulating osteoprotegerin levels and improved secondary hyperparathyroidism. The observed effects were the opposite of those expected from previous in vitro studies. Osteoprotegerin may mediate and/or modify the effect of active vitamin D therapy in dialysis patients. PMID- 16047648 TI - Normalization of reversed bio-intact-PTH(1-84)/intact-PTH ratio after parathyroidectomy in a patient with severe secondary hyperparathyroidism. AB - The conventional intact-PTH assay detects both (1-84)-PTH and C-terminal fragments. The newer PTH assays, bio-intact-PTH assay and whole-PTH assay, use an antibody that binds only if the first amino acid is present, making it specific for the complete molecule, (1-84)-PTH. Thus, the intact-PTH concentrations are theoretically higher than bio-intact-PTH concentrations, and the ratio of bio intact-PTH/intact-PTH is usually less than 1. These findings are observed in normal subjects and patients with primary and secondary hyperparathyroidism. Here we present a hemodialysis patient with severe secondary hyperparathyroidism who was found to have abnormally higher plasma bio-intact-PTH concentrations than intact-PTH concentrations, and the abnormally high biointact-PTH/intact-PTH ratio improved after parathyroidectomy (PTx). The patient was a 67-year-old man on maintenance hemodialysis since 1995. Since 2003, he was found to have high plasma intact-PTH concentrations and two swollen parathyroid glands in the neck. PTx with forearm autograft was performed in October 2003. Before PTx, an abnormally high ratio of bio-intact-PTH/intact-PTH was detected (840 pg/ml/770 pg/ml, > 1), while the same ratio was improved to normal range (100 pg/ml/200 pg/ml, < 1). Recently, a few patients with parathyroid carcinoma have been found to have higher (1-84)-PTH concentrations than intact-PTH concentrations with abnormally high (1-84)-PTH/intact-PTH ratio. Moreover, a new molecular form of PTH distinct from (1-84)-PTH was detected in these patients. We speculate that the resected parathyroid gland in our patient might have produced a new molecular form of PTH that was less well detected by the conventional intact-PTH assay. PMID- 16047649 TI - Failure of rituximab to treat a lupus flare-up with nephritis. AB - The autoantibodies secreted by B lymphocytes have recently been shown to play an important role in autoimmune disease. B lymphocyte depletion by rituximab, a monoclonal anti-CD20 antibody, has been introduced for the treatment of several autoimmune disorders. Few reports have underlined its potential use for the treatment of systemic lupus erythematosus (SLE). We report here the occurrence of extracapillary glomerulonephritis associated with a thrombotic event shortly after rituximab treatment for a lupus flare-up in a patient with anticardiolipin antibodies. This observation suggests that rituximab alone may be insufficient to control severe SLE with glomerulonephritis and should therefore be used with caution in patients with this condition. PMID- 16047650 TI - Increasing incidence of focal segmental glomerulosclerosis. PMID- 16047651 TI - Mizoribine decreases urinary protein excretion in two patients with IgA nephropathy. PMID- 16047652 TI - Can minimal change nephrotic syndrome superimposed on diabetic nephropathy be diagnosed? PMID- 16047653 TI - An interview with Adrian Hochstadt. Interview by R. Scott Nolen. PMID- 16047654 TI - Questions methods and conclusions in farm animal welfare survey. PMID- 16047655 TI - What is your diagnosis? Degenerative joint disease of the hip. PMID- 16047656 TI - Theriogenology question of the month. Urospermia. PMID- 16047657 TI - Animal behavior case of the month. Behavioral seizures. PMID- 16047658 TI - Size and organization of private veterinary practices, 2003. PMID- 16047659 TI - Frequency of urinary tract infection among dogs with pruritic disorders receiving long-term glucocorticoid treatment. AB - OBJECTIVE: To determine frequency of urinary tract infection (UTI) among dogs with pruritic disorders that were or were not receiving long-term glucocorticoid treatment. DESIGN: Observational study. ANIMALS: 127 dogs receiving glucocorticoids for > 6 months and 94 dogs not receiving glucocorticoids. PROCEDURE: Bacterial culture of urine samples was performed in dogs receiving long-term glucocorticoid treatment, and information was collected on drug administered, dosage, frequency of administration, duration of glucocorticoid treatment, and clinical signs of UTI. For dogs not receiving glucocorticoids, a single urine sample was submitted for bacterial culture. RESULTS: Multiple (2 to 6) urine samples were submitted for 70 of the 127 (55%) dogs receiving glucocorticoids; thus, 240 urine samples were analyzed. For 23 of the 127 (18.1%) dogs, results of bacterial culture were positive at least once, but none of the dogs had clinical signs of UTI. Pyuria and bacteriuria (present vs absent) were found to correctly predict results of bacterial culture for 89.9% and 95.8% of the samples, respectively. Type of glycocorticoid, dosage, frequency of administration, and duration of treatment were not associated with frequency of UTI. None of the urine samples from dogs not receiving glucocorticoids yielded bacterial growth. The frequency of UTI was significantly higher for dogs treated with glucocorticoids than for dogs that had not received glucocorticoids. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that dogs receiving long-term glucocorticoid treatment have an increased risk of developing a UTI. On this basis, we recommend that urine samples be submitted for bacterial culture at least yearly for such dogs. PMID- 16047660 TI - Evaluation of a rebound tonometer for measuring intraocular pressure in dogs and horses. AB - OBJECTIVE: To compare intraocular pressure (IOP) measurements obtained with a rebound tonometer in dogs and horses with values obtained by means of applanation tonometry and direct manometry. DESIGN: Prospective study. ANIMALS: 100 dogs and 35 horses with clinically normal eyes, 10 enucleated eyes from 5 dogs, and 6 enucleated eyes from 3 horses. PROCEDURES: In the enucleated eyes, IOP measured by means of direct manometry was sequentially increased from 5 to 80 mm Hg, and IOP was measured with the rebound tonometer. In the dogs and horses, results of rebound tonometry were compared with results of applanation tonometry. RESULTS: For the enucleated dog and horse eyes, there was a strong (r2 = 0.99) linear relationship between pressures obtained by means of direct manometry and those obtained by means of rebound tonometry. Mean +/- SD IOPs obtained with the rebound tonometer were 10.8 +/- 3.1 mm Hg (range, 5 to 17 mm Hg) and 22.1 +/- 5.9 mm Hg (range, 10 to 34 mm Hg) for the dogs and horses, respectively. Mean IOPs obtained with the applanation tonometer were 12.9 +/- 2.7 mm Hg (range, 8 to 18 mm Hg) and 21.0 +/- 5.9 mm Hg (range, 9 to 33 mm Hg), respectively. Values obtained with the rebound tonometer were, on average, 2 mm Hg lower in the dogs and 1 mm Hg higher in the horses, compared with values obtained with the applanation tonometer. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the rebound tonometer provides accurate estimates of IOP in clinically normal eyes in dogs and horses. PMID- 16047661 TI - Use of a percutaneous atrial septal occluder device for complete acute occlusion of an intrahepatic portosystemic shunt in a dog. AB - A 3-month-old sexually intact male German Shepherd Dog was evaluated because of signs of depression, ataxia, and collapse. Clinicopathologic abnormalities included low serum BUN and albumin concentrations and high serum liver enzyme activities and plasma ammonia and serum bile acids concentrations. Abdominal ultrasonography revealed an intrahepatic portosystemic shunt (PSS). The dog was anesthetized; via a transjugular approach, guidewires and catheters were directed with fluoroscopic guidance to locate the shunt and determine its anatomic features. Minimal changes in portal vein pressure during temporary shunt balloon occlusion enabled complete shunt attenuation, which was performed by use of a self-expanding septal occlusion device that is typically used for treatment of atrial septal defects in humans. Following initial misplacement of the device, the procedure was repeated successfully 2 months later and resulted in complete shunt occlusion. One year after this second procedure, the dog was clinically normal and serum bile acids concentration was within reference limits. In certain dogs with intrahepatic PSSs, treatment with minimally invasive interventional techniques involving fluoroscopy may reduce the morbidity and mortality rates associated with more invasive surgical procedures. PMID- 16047662 TI - Volvulus of the colon in four dogs. AB - Four dogs were examined because of vomiting of 7 to 48 hours' duration. Gas distended segments of intestine were identified radiographically in all dogs, but the affected portion of the intestinal tract could not always be identified as the colon. Volvulus of the colon was diagnosed during surgery in all 4 dogs. Gastrocolopexy was performed following derotation of the colon in 3 of the dogs. In 1 dog, a colectomy and an ileorectal anastomosis were performed. All 4 dogs survived. Volvulus of the colon should be considered as a cause of vomiting of short duration in dogs for which there is radiographic evidence of intestinal dilatation. PMID- 16047663 TI - Evaluation of nonsurgical treatment of atlantoaxial subluxation in dogs: 19 cases (1992-2001). AB - OBJECTIVE: To evaluate long-term neurologic outcome in dogs with atlantoaxial subluxation (AAS) that were treated nonsurgically with a cervical splint. DESIGN: Retrospective study. ANIMALS: 19 dogs with AAS and managed with a cervical splint. PROCEDURE: Medical records from 2 university hospitals were reviewed. Information pertaining to trauma, duration of clinical signs prior to admission, medical treatments prior to admission, results of neurologic and physical examinations at the time of admission, results of laboratory testing, results of diagnostic imaging, neurologic status at the time of discharge, duration of time the cervical splint was used for treatment, and neurologic status at the time of splint removal and at a final reexamination was extracted from the medical records. Long-term outcome was defined as neurologic status greater than or equal to 1 year after splint removal. Factors associated with a good or poor long-term outcome were determined. RESULTS: A good final outcome was reported in 10 of 16 dogs. Median duration of clinical signs prior to referral was 30 days; dogs that were affected < or = 30 days were significantly more likely to have a good long term outcome, compared with dogs affected > 30 days. The neurologic grade at admission, radiographic appearance of the dens, age at onset of clinical signs, and history were not associated with outcome. CONCLUSIONS AND CLINICAL RELEVANCE: Nonsurgical management of AAS by use of a cervical splint is a viable treatment modality for young dogs with a first episode of acute-onset clinical signs, regardless of the severity of neurologic deficits at admission. PMID- 16047664 TI - Bone marrow necrosis in dogs: 34 cases (1996-2004). AB - OBJECTIVE: To identify the incidence, potential causes, and clinical and clinicopathologic features of bone marrow necrosis in dogs. DESIGN: Retrospective study. ANIMALS: 34 client-owned dogs. PROCEDURES: Reports of cytologic examinations of bone marrow specimens performed between 1996 and 2004 were reviewed. All reports that indicated the presence of necrosis, stromal disruption, phagocytic macrophages, individual cell necrosis, or myelofibrosis were evaluated further. RESULTS: Of 609 reports of bone marrow evaluations performed during the study period, 34 (5.6%) had evidence of bone marrow necrosis. Nine dogs had no evidence of associated diseases or drug or toxin exposure, and 25 dogs had associated disease conditions or drug exposures. All 9 dogs with idiopathic bone marrow necrosis were anemic (mean Hct, 14%), but only 3 had neutropenia, and 3 had thrombocytopenia. All 9 had myelofibrosis. Of the 25 dogs with associated disease conditions or drug exposures, only 14 (56%) had anemia (mean Hct, 33%). In addition, 14 (56%) had neutropenia and 18 (72%) had thrombocytopenia. Only 10 (40%) had myelofibrosis. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that bone marrow necrosis may be common in dogs with hematologic disorders. In most dogs, bone marrow necrosis was associated with an underlying disease condition or drug exposure, but idiopathic bone marrow necrosis was also identified. Disease conditions that should increase suspicion of possible bone marrow necrosis include sepsis, lymphosarcoma, and systemic lupus erythematosus; drug exposures that should increase suspicion of possible bone marrow necrosis include chemotherapeutic agents, phenobarbital, carprofen, metronidazole, and mitotane. PMID- 16047665 TI - Signalment and clinical features of diskospondylitis in dogs: 513 cases (1980 2001). AB - OBJECTIVE: To determine signalment, clinical features of the disease, and treatment in dogs with diskospondylitis. DESIGN: Case-control study. ANIMALS: 513 dogs with diskospondylitis (cases) and 236,109 canine hospital accessions (controls) from 12 veterinary teaching hospitals. PROCEDURE: Information retrieved from the medical records of 123 dogs with diskospondylitis at the Louisiana State University veterinary teaching hospital between 1980 and 2001 included sex, age, breed, primary complaint, neurologic status, location of lesions, causative organism, treatment, and outcome. The signalment of 390 additional cases from 11 other veterinary teaching hospitals was accessed from the Veterinary Medical Database. Comparisons were made with controls from the same time periods. RESULTS: Male dogs were twice as likely as female dogs to be affected (odds ratio [OR], 2.0; 95% confidence interval [CI], 1.7 to 2.4). Dogs were significantly more likely to be affected as age increased. Purebred dogs, especially Great Danes, were more likely than mixed-breed dogs to be affected (OR, 73; CI, 4.3 to 12.6). For dogs from Louisiana State University, Staphylococcus spp, Brucella spp, Streptococcus spp, and Escherichia coli were isolated most often; multiple organisms were detected via microbial culture in 11 dogs. The mean duration of treatment was 53.7 weeks. CONCLUSIONS AND CLINICAL RELEVANCE: Male dogs, older dogs, and Great Danes appeared more likely to be affected with diskospondylitis than female dogs, dogs < 1 year of age, and mixed breed dogs, respectively. Long-term administration of antimicrobial drugs for treatment of diskospondylitis may be expected. Identification of the causative organism and early treatment are recommended. PMID- 16047666 TI - Evaluation of plasma catecholamine and serum cortisol concentrations in horses with colic. AB - OBJECTIVE: To evaluate plasma epinephrine and norepinephrine concentrations and serum cortisol concentration in horses with colic and assess the relationship of these variables with clinical signs, routinely measured clinicopathologic variables, and outcome in affected horses. DESIGN: Prospective observational study. ANIMALS: 35 horses with colic. PROCEDURE: Blood samples were collected within 30 minutes of arrival at the veterinary hospital from horses referred because of colic. Plasma and serum samples were analyzed for cortisol, epinephrine, norepinephrine, lactate, and electrolyte concentrations and acid base variables. Heart rate at admission and outcome (survival or nonsurvival) were recorded. Univariate logistic regression was used to calculate crude (unadjusted) odds ratios and 95% confidence intervals. RESULTS: Of the 35 horses with colic, 26 survived. Higher plasma epinephrine, plasma lactate, and serum cortisol concentrations were significantly associated with increased risk of nonsurvival, but plasma norepinephrine concentration was not associated with outcome. Plasma epinephrine concentration was significantly correlated with heart rate (r = 0.68), plasma lactate concentration (r = 0.87), blood pH (r = -0.83), anion gap (r = 0.74), and base excess (r = -0.81). CONCLUSIONS AND CLINICAL RELEVANCE: The risk of death appears to be greater in colic-affected horses with high circulating concentrations of epinephrine and cortisol. The correlation of epinephrine with other biochemical markers of illness severity and with heart rate indicates that the degree of sympathetic activation in horses with colic can be inferred from routinely measured variables. PMID- 16047667 TI - Evaluation of electroacupuncture treatment of horses with signs of chronic thoracolumbar pain. AB - OBJECTIVE: To evaluate use of electroacupuncture for treatment of horses with signs of chronic thoracolumbar pain. DESIGN: Prospective study. ANIMALS: 15 horses with signs of chronic thoracolumbar pain. PROCEDURE: Horses were randomly allocated to 1 of 3 treatment groups. Horses in group 1 received electroacupuncture stimulation (once every 3 days for 5 treatments), those in group 2 received phenylbutazone (2.2 mg/kg [1 mg/lb], PO, q 12 h, for 5 days), and those in group 3 received saline (0.9% NaCl) solution (20 mL, PO, q 12 h, for 5 days). Thoracolumbar pain scores (TPSs) were evaluated before (baseline) and after each treatment. RESULTS: Mean +/- SE TPSs in horses receiving phenylbutazone or saline solution did not change significantly during the study. After the third treatment, mean +/- SE TPS (2.1 +/- 0.6) in horses receiving electroacupuncture stimulation was significantly lower than baseline (6.0 +/- 0.6) TPS. Mean +/- SE TPSs in horses receiving electroacupuncture stimulation were significantly lower than baseline TPSs and TPSs in horses receiving phenylbutazone or saline solution after the third treatment to 14 days after the last treatment. CONCLUSIONS AND CLINICAL RELEVANCE: TPSs are useful for evaluating the efficacy of various analgesic methods used for treatment of thoracolumbar pain in horses. Electroacupuncture was effective for treatment of chronic thoracolumbar pain in horses. Results provided evidence that 3 sessions of electroacupuncture treatment can successfully alleviate signs of thoracolumbar pain in horses. The analgesic effect induced by electroacupuncture can last at least 2 weeks. Phenylbutazone administered PO did not effectively alleviate signs of thoracolumbar pain in horses in this study. PMID- 16047668 TI - Abomasal impaction in Holstein-Friesian cows: 80 cases (1980-2003). AB - OBJECTIVE: To determine clinical examination findings, clinicopathologic abnormalities, and outcome of treatment in dairy cattle with abomasal impaction. DESIGN: Retrospective study. ANIMALS: 80 lactating Holstein-Friesian cows > or = 2 years old. PROCEDURE: Medical records of cattle with abomasal impaction admitted between 1980 and 2003 were retrieved, and data were extracted. RESULTS: All cows were reported to have decreased food intake; concurrent diseases were identified in 54 (68%). Seventeen cows did not have detectable ruminal motility, but physical examination findings were nonspecific and variable. In general, cattle had mild hypocalcemia, hyperbilirubinemia, and hyperglycemia, but serum potassium and chloride concentrations were typically within reference limits. Fifty-five (69%) cattle had impaction of the pyloric antrum alone, and 25 (31%) had impaction of the abomasal body and pyloric antrum. Right flank laparotomy and abomasal massage were performed in 73 cattle. After surgery, 54 (74%) cattle received 3 to 4 L of mineral oil, PO, daily for 1 to 5 days. Short-term (ie, discharged from the hospital) survival rate was significantly higher for cows with impaction of the pyloric antrum alone (42/45 [93%]) than for cows with impaction of the body and antrum (12/24 [50%]). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that physical examination findings and results of serum biochemical analyses do not facilitate the diagnosis of abomasal impaction in lactating Holstein cows and that exploratory right flank laparotomy is necessary to make the diagnosis. Abomasal impaction should be considered as a differential diagnosis for inappetence and poor milk production in lactating dairy cows. PMID- 16047669 TI - [Studying the mechanisms of ladasten action]. AB - The effect of N-(2-adamantyl)-N-(p-bromophenyl)amine (ladasten) on the behavior of MR and MNRA inbred rats with different emotional-stress reaction (ESR) phenotypes was studied in the conventional open-field test. Ladasten (30 mg/kg, i.p.) produced anxiolytic effect in MR rates, while not influencing MNRA animals. At the same time, a model stress induced disorder in the regulation of 3 H diazepam binding to the benzodiazepine site of GABA(A) receptor in MR (but not in MNRA) rats. Ladasten administered in the anxiolytic dose prevented stress-induced changes in reception at this site. PMID- 16047670 TI - [Antidepressant effect of L-tryptophan in male rats with dysbalance thyreoid hormones]. AB - The activity of L-tryptophan was studied on young male rats in depressive state caused by thyreoid hormone dysbalance. The immobilization time in the Porsolt forced swim test in thyreoidectomized rats was significantly increased in comparison to the intact control. The replacement therapy with triiodothyronin (T3) reduced the level of depressed behavior to the initial value. Chronic administration of L-tryptophan significantly decreased the total immobilization time in all the test groups, irrespective of the hormonal state. However, the effect of L-tryptophan was more pronounced in T3-treated rats. PMID- 16047671 TI - [Comparative study of the antidepressant and anxiolytic activity of fluoxetine and tianeptine]. AB - The activity of fluoxetine (prozac) and tianeptine (coaxil) was studied in outbred male rats under Porsolt and S. Nomura modified forced swim tests. The antidepressant effect of tianeptine was much more pronounced that that of fluoxetine. In the conflict situation test, fluoxetine produced anxiogenic action. In contrast, tianeptine decreased (albeit not reliably) the anxiety. In combination with bicuculline (GABAA receptor blocker), the anxiolytic action was more pronounced for both antidepressants, which was manifested by a significant increase in the number of punished water takes. A neural network explaining the observed behavior is proposed, which includes GABA-ergic intemeurons inhibiting serotonin release from serotoninergic terminals. PMID- 16047672 TI - [Effect of oxytocin on the activity of neurons in posterior hypothalamic regions of stressed rats]. AB - The dynamics of pulse activity in the cells of posterior hypothalamic regions was studied in acute experiments on white rats treated with oxytocin. Intravenous injections of the drug induced dose-dependent reorganization of the neuron discharge both in intact rats and in animals with obvious emotional sexual stress. The dose-dependent effect was observed in various latent periods of the neuron response and was manifested by changes in the character of pulsation. The higher the background frequency of cell pulses, the less pronounced was the neuron response to the drug. In some cases, the drug intensified the burst activity and produced regulation of the discharge rhythm. PMID- 16047673 TI - [Comparative study of the physical dependence on proline analog of dermorphin and morphine]. AB - On the model of acute physical dependence, naloxone treatment of animals dependent on D-Pro6 peptide ([D-Pro6]DM) (a demorphin analog) led to tremor, shaking, convulsions, and rare jumps typical of morphine-dependent animals. The variety and intensity of reactions depended on the naloxone dose and the interval between naloxone and peptide injections. In the state of reject syndrome upon peptide and morphine subchronic treatments according to identical schedules, the symptoms in [D-Pro6]DM dependent animals were much less pronounced than in morphine-dependent ones. PMID- 16047674 TI - [The role of timing factors for the effect of caffeine on heart rhythm variability in healthy humans]. AB - Caffeine upon single administration optimizes the indices of heart rhythm variability in young humans and enhances parasympathotonia. The expression of this reaction depends on the daytime (being more pronounced in the morning than in the evening) and on the chronotype (morningness) of volunteers. PMID- 16047675 TI - [The effect of hypoxen on the stress-induced ulcerogenesis in rats]. AB - Immobilization stress induced ulcerous damage in the stomach mucosa of test rats. Prophylactic treatment with hypoxen in a daily dose of 50 mg/kg over a 5-day period decreased the number of ulcers and the total area of erosive injury in the mucous membrane, decreased the lipid peroxidation rate, and reduced the involution of thymus gland. This drug action can be classified as gastroprotective. PMID- 16047676 TI - [Effect of ladasten on reproduction function in male rats]. AB - The effect of ladasten (bromantan) on the development of emotional stress and reproduction activity was studied in male rats. Ladasten administration led to restoration of the stress-violated orientational reactions and body weight, activation of the spermatogenesis (increase in the number and mobility of spermatozoas), normalization of the fertilizing capacity of spermatozoas, and reduction in the loss of embryos. PMID- 16047677 TI - [Effect of the new antimigraine drug serotonin receptor antagonist tropoxin on platelet aggregation]. AB - The effect of the new antimigraine drug tropoxin - the serotonin receptor (5-HT2) antagonist - on the human platelet aggregation in vitro induced by ADP (1 x 10( 5) M) and epinephrine (2.5 x 10(-6) M) was studied. Tropoxin reliably inhibited the ADP-induced platelet aggregation in a concentration range of 0.01 - 7 mg/ml. A significant inhibition effect with respect to the epinephrine-induced platelet aggregation was observed in a drug concentration range of 2 - 7 mg/ml, although a reliable antiaggregant activity was also observed below 2 mg/ml. A bolus administration of tropoxin (10 mg/kg) in rabbits decreased the ADP-induced platelet aggregation ex vivo by a factor of 1.2 - 1.4. The effect appeared 45 min after treatment and was observed during subsequent 30 min. PMID- 16047678 TI - [The effect of a combined vitamin--antioxidants preparations on hemostasis in rats with experimental hyperperoxidation]. AB - Experiments on a group of 308 rats showed that prooxidant (lead acetate) introduced per os with daily meals decreases the antioxidant potential (increases lipid peroxidation, LPO) and reduces tolerance with respect to thrombin (increases the level of intravascular blood coagulation markers). For the same treatment on the background of a vitamin - antioxidants preparations (complivit or selmevit), the level of changes was significantly decreased and the resistance to thrombin was increased. Selmevit (containing selenium) was more effective. It is concluded that complex vitamin--antioxidants preparations can be used for correcting hemostatic changes in cases of hyperoxidation. PMID- 16047679 TI - [Mexidol corrects model post-reanimation changes in cerebral lipid metabolism]. AB - The lipid spectrum of brain tissues in various stages of post-reanimation period was studied in rats upon clinical death modeled by cardiovascular fascicle ligation according to V. G. Korpachev. Reproduction of same model on the background of mexidol was characterized by (i) an increase in phosphatidylserine, (ii) normalization of the relative content of phosphatidylethanolamine and cholesterol, (iii) stabilization of the content of phosphatidylcholine, sphyngomyelin, lysophosphatidylethanolamine, and free fatty acids, and (iv) the absence of lysophosphatidylserine in late post-reanimation period (30 days) upon mexidol reperfusion. Mexidol also favored the development of protective and adaptive reactions on the level of lipid components of cerebral cell membranes in the early period upon reperfusion and prevented from the development of irreversible changes in cerebral phospholipid metabolism in the late post reanimation period. PMID- 16047680 TI - [Effect of phenibut on the behavior of experimental animals under conditions of voluntary chronic alcoholism]. AB - The effect of phenibut on the locomotor and orientation-research activity, as well as on the alcohol and food motivation, was studied on experimental animals under conditions of voluntary chronic alcoholism. Phenibut decreased the manifestations of alcohol-induced behavioral disorders and reduced alcohol motivation. PMID- 16047681 TI - [Experimental justification of the use of benzimidazole derivatives as antiasthenic agents after craniocerebral trauma]. AB - Experimental administration of benzimidazole derivatives upon model craniocerebral trauma favored rapid normalization of the behavior of hypoxia sensitive animals and prevented from excess activation of lipid peroxidation processes in the brain. It is recommended to use benzimidazole derivatives for the control of free-radical processes and the correction of psychopathologic disorders in cases of post-trauma cerebroasthenia. PMID- 16047682 TI - [Evaluation of anti-seasick properties of pyrroxan and dimenhydrinate under conditions of maximal and submaximal statokinetic load in humans]. AB - Pyrroxan and dimenhydrinate exhibit comparable anti-seasick effects in the test involving maximal and submaximal statokinetic load. Pyrroxan somewhat more significantly increases the vestibular stability, predominantly in humans with inherently high and medium stability, whereas dimenhydrinate is also effective in humans with low resistance to seasick. Pyrroxan primarily decreases the statokinetic (somatic) manifestations (dizziness, defensive movements, nystagmus), while dimenhydrinate mostly abolishes vegetative manifestations (hyperhydrosis, nausea, vomiting, fever sensation). Thus, the two drugs produce approximately equal anti-seasick action, while differing in the point of application (somatic versus vegetative). PMID- 16047683 TI - [The effect of tofisopam and tinctura leonuri on the color-discrimination function in young humans]. AB - Enhanced anxiety state is accompanied by limitation of the color-discrimination function of retina in young humans. Chronic administration of tofisopam (grandaxin) or tinctura leonuri decreased anxiety and significantly improved the color discrimination function of retina with respect to all four colors studied. It is suggested that this improvement is related to the drug action upon the GABAergic processes both in the retina and in the related cerebral structures. PMID- 16047684 TI - [Morphological evaluation of the process of aseptic skin ulcer healing under the action of prolidoxide ointment]. AB - The effect of prolidoxide ointment on the healing of aseptic skin ulcers in rats was evaluated in terms of skin morphology. Prolidoxide exhibited more pronounced healing action than the analogous (with respect to the composition of natural components) reference ointment ("Wundahyl"). It is recommended to use prolidoxide for accelerated skin surface healing in the first and second stage of the ulceration process. PMID- 16047685 TI - [Finding cholinesterase in endotheliocytes: cholinesterase inhibition by organophosphorus compounds leads to endotheliocyte deformation]. AB - Toxic action of some organophosphorus cholinesterase inhibitors (including phosphacol) on the state of microvessels was studied in rats. Cholinesterase was found in endotheliocytes and it was established that phosphacol inhibited this enzyme. This was one of the factors responsible for deformation of the luminal relief of the vessels and for violation of the blood microcirculation. The expression of these effects was different for various organophosphorus compounds. PMID- 16047686 TI - [Daphnia magna Straus: a new model for evaluating the antioxidant acton of water soluble preparations in vivo]. AB - Hydrobionta species of Daphnia magna Straus were used as a test objects for in vivo evaluation of the antioxidant activity of three hydrophilic thiol compounds: reduced gluthatione, unithiol, and cysteine. These compounds exhibited significant differences in activity under oxidative stress conditions, in the dynamics of observed effects, and in the probability of inversion from anti- to pro-oxidant action. The main advantage of the proposed test objects in comparison to the conventional in vitro experiments (where the antioxidant effect is evaluated over a period of time from several minutes to several hours) is that the development of drug activity (pro- and antioxidant effects) can be monitored over a prolonged period of time (up to several days). In comparison to the tests on mammals, the new method is much simpler and allows the entire antioxidant protection (rather than separate systems) to be evaluated. It is recommended to use Daphnia magna Straus species for comparative evaluation of the antioxidant action of water-soluble preparations in vivo. PMID- 16047687 TI - [70 years of the pharmacology department of the Dagestan State Medical Academy]. PMID- 16047688 TI - [Epiphyseal hormone melatonin: a new nootrope?]. PMID- 16047689 TI - Assessing four decades of Medicare, Medicaid. AB - Medicare and Medicaid were born 40 years ago, offering the first federally funded safety net for senior citizens, the uninsured and the disabled. While President Theodore Roosevelt, left, first proposed a national health insurance program in 1912, it took decades of political infighting before a program could be created. A look at the past, present and possible future of these important programs begins on Page 6. PMID- 16047690 TI - Malpractice not the issue. Study finds other factors are driving medical costs. PMID- 16047691 TI - Same outcomes, different costs. Study finds bypass surgery less costly in Canada. PMID- 16047693 TI - LifePoint adds five HCA hospitals. PMID- 16047692 TI - Pa. releases infection data. PMID- 16047694 TI - Compensating nurses. Geography, intangibles factor into pay disparities. PMID- 16047695 TI - More dollars for doctors. Proposed formula may increase Medicare payments. PMID- 16047696 TI - Wellpoint settles with docs. 198 million dollars closes long-running civil lawsuit. PMID- 16047697 TI - Off the hook. Former McKesson CFO acquitted in fraud case. PMID- 16047698 TI - States of relief. Appeals board says five states won't have to repay. PMID- 16047699 TI - Learning from history. Old adage about not repeating past mistakes actually holds some wisdom. PMID- 16047700 TI - Holding steady. Specialists are still seeing biggest paychecks, but raises are flat, and some generalists are gaining ground, physician compensation survey shows. PMID- 16047701 TI - Sign of the times. Medicare is a great program, but even a good thing should be updated. PMID- 16047702 TI - Modern perspective. Medicare milestone offers perfect time to regroup. PMID- 16047703 TI - Payment reform needed. Looming cuts may exacerbate access crisis for seniors. PMID- 16047704 TI - Mired in the past. Medicare, Medicaid need a big dose of modernization. PMID- 16047705 TI - Researching the truth. Debunking Medicaid and Medicare myths. PMID- 16047706 TI - Securing the hospital food chain. Food safety emerges as another homeland security challenge as hospitals, vendors update disaster plans and tighten procedures. PMID- 16047707 TI - Unproven techniques in allergy diagnosis. AB - Mainstream allergy diagnosis and treatment is based on classical allergy testing which involves well-validated diagnostic methods and proven methods of treatment. By contrast, a number of unproven tests have been proposed for evaluating allergic patients including cytotoxic food testing, ALCAT test, bioresonance, electrodermal testing (electroacupuncture), reflexology, applied kinesiology a.o. There is little or no scientific rationale for these methods. Results are not reproducible when subject to rigorous testing and do not correlate with clinical evidence of allergy. Although some papers suggest a possible pathogenetic role of IgG, IgG4 antibody, no correlation was found between the outcome of DBPCFC and the levels of either food-specific IgG or IgG4, nor was any difference seen between patients and controls. The levels of these and other food-specific immunoglobulins of non-IgE isotype reflect the intake of food in the individual and may thus be a normal and harmless finding. The so-called "Food Allergy Profile" with simultaneous IgE and IgG determination against more than 100 foodstuffs is neither economical nor useful for diagnosis. DBPCFC must be the reference standard for food hypersensitivity and any new test must be validated by it. As a result, all these unproven techniques may lead to misleading advice or treatments, and their use is not advised. PMID- 16047708 TI - Reducing the risk of anaphylaxis during anaesthesia: guidelines for clinical practice. AB - These guidelines represent a consensus of experts in the field of immediate hypersensitivity reactions occurring during anaesthesia. They were based on international science, and implemented in France under the auspices of the French Society for Anaesthesia and Intensive Care (SFAR: Societe Francaise d'Anesthesie et de Reanimation). Their aim was to provide the most valid, widely accepted, effective and easily teachable guidelines that current knowledge, research and experience can provide. This paper presents the main extracts of these recommendations with the most relevant clinical implications. PMID- 16047709 TI - Comparative analysis of the bronchodilator response measured by impulse oscillometry (IOS), spirometry and body plethysmography in asthmatic children. AB - BACKGROUND: Asthma is common among young children. The assessment of respiratory resistance by the impulse oscillometry system (IOS), based on the superimposition of respiratory flow by short-time impulses, requires no patient active collaboration. AIM: We evaluated the baseline repeatability and bronchodilator response of IOS indices in preschool children, their correlation with spirometry and whole body plethysmography, and differences between atopic and nonatopic children. PATIENTS AND METHODS: Thirty-three asthmatic children (3-6 yrs.) underwent IOS measurement (R5rs, R20rs and X5rs) by triplicate at the baseline, after placebo and after salbutamol inhalation. Spirometry (FEV1) and whole body plethysmography (sRaw) were made at the baseline and after salbutamol. Baseline within-test (coefficient of variation: CV%) and between-test repeatability (baseline-placebo) were addressed. Bronchodilator response was evaluated by the SD index (change in multiples of the between-test repeatability). RESULTS: Baseline repeatability for R5rs was 4.1%. Its values decreased by 2SD after salbutamol inhalation, and correlated with FEV1 and sRaw at both, baseline (r= 0.51 and r=0.49) and post-salbutamol (r=-0.63 and r=0.54). A trend towards correlation between salbutamol-induced changes in R5rs and in sRaw (r=0.33) was observed. Atopic and non-atopic children showed no differences in lung function. CONCLUSION: IOS was well accepted by young asthmatic children and provided reproducible and sensitive indices of lung function. Resistance values obtained by IOS at low frequency (R5rs) were reproducible and correlated with spirometry and plethysmographic values. PMID- 16047710 TI - Characterization of T-cell clones specific to ovomucoid from patients with egg white allergy. AB - BACKGROUND: Allergic reactions to foods are specific problems for infants and young children. Ovomucoid (OM) is one of the major allergens found in egg-white. We previously established several T-cell clones (TCCs) specific to OM in non polarizing conditions from 4 patients (TM and YN are immediate-type, IH and YT are non-immediate-type) with egg-white allergy. We characterized their reactive epitopes, antigen-presenting molecules (HLA class II), and usage of TCR alpha and beta genes and the CDR3 loop sequence. OBJECTIVE: The objective of this study was to characterize these seven clones (TM 1.3, TM1.4,YN 1.1, YN1.5, IH3.1, IH3.3 and YT6.1) for cytokine production patterns and cell-surface-marker phenotypes. METHODS: We measured the production of cytokines, namely interleukin (IL)-4, IL-5 and interferon-gamma (IFN-gamma) by stimulation with ovomucoid peptides and stained intracellular IL-4 and IFN-gamma, and determined cell-surface markers using anti-interleukin-12 receptor (IL-12R) beta1, anti-IL-12Rbeta2 and anti interleukin-18 receptor alpha (IL-18Ralpha). RESULTS: Most TCCs secreted both IL 4 and IFN-gamma in response to the OM peptide mixture, but the secretion patterns were variable; an IFN-gamma dominant pattern was seen in IH3.1 andYT6.1, an IFN gamma>IL-4 pattern in TM1.3 and TM1.4, an IL-4> IFN-gamma pattern in YN1.5. In intracellular IFN-gamma and IL-4 staining, IFN-gamma single-positive cells were predominant in TM1.3, TM1.4, IH3.1 and YT6.1 and IFN-gamma and IL-4 double positive cells were predominant in YN1.1, YN1.5 and IH3.3. All TCCs were IL 12Rbeta1-positive, and TM1.3, IH3.1, IH3.3 and YT6.1 were both IL-12Rbeta2- and IL-18Ralpha-positive. TM1.4 and YN1.1 were both IL-12Rbeta2- and IL-18Ralpha negative. Based on these results, TM1.3 and TM1.4, IH3.1 and YT6.1 had a predominantly Th1 character and YN1.1, YN1.5, and IH3.3 possessed a predominantly Th0 character. CONCLUSIONS: The phenotypes of TCCs were not in accordance with their clinical manifestations. TCCs established from patients with immediate-type hypersensitivity had either the Th1 or Th0 phenotype as well as those with non immediate-type hypersensitivity. PMID- 16047711 TI - Correlation between pollen counts and symptoms in two different areas of the Iberian Peninsula: Cordoba (Spain) and Evora (Portugal). AB - This work was designed to analyse the symptoms of pollen allergy in two different cities of the Iberian Peninsula, Cordoba and Evora. Fifteen patients with pollinosis in Cordoba and twenty-seven in Evora were included in this study. Rhinitis symptoms scores were recorded in both cities in order to correlate these data with those of pollen counts. Linear regression analyses were performed in order to find out the contribution of different taxa to the sign of allergy symptoms in the allergenic population. Skin prick tests were also considered. A high and significant correlation was found for Poaceae (0.89), Olea (0.73) and Plantago (0.56) in Cordoba and Olea (0.53) in Evora. Results also suggest that the incidence of the different pollen types in allergy patients is higher in Cordoba than in Evora. A rural lifestyle in Evora that confers a better protection from environmental agents in addition to a less contaminated atmosphere could be the cause. PMID- 16047712 TI - The effects of environmental pollution on the respiratory system of children in western Macedonia, Greece. AB - The indoor and outdoor environmental pollution effects on the respiratory system of 3,559 children aged 9-12 were studied. It was a cross-sectional and interlocal (geographical differentiation) study. The research was conducted during the period between 2000-2001 in five cities of Western Macedonia and more particularly: 1046 children from Ptolemaida, 1249 children from Kozani, 466 from Florina, 419 from Kastoria and 379 from Grevena. The study was performed by means of a questionnaire for the detection of respiratory diseases during childhood, plus spirometry and rhinomanometry measurements. The diachronic quantitative analysis of environmental pollutants was conducted by The Laboratory of Physics of the Atmosphere of the Aristotle University of Thessaloniki. The environmental pollution was found to have a detrimental effect on the respiratory system of children, mainly attributable to the occurrence of rhinitis and infectious bronchitis. The highest prevalence of rhinitis (40.3%) and infectious bronchitis (12.1%) was observed in Ptolemaida, which is a highly polluted region, whereas the lowest (21.2% and 6.7%, respectively) was seen in Grevena, a non-polluted area. As for the indoor pollution, maternal smoking was found to increase the prevalence of respiratory problems in children. Finally, the father's educational level and a past history of nursery school attendance increase the prevalence of respiratory diseases during childhood. PMID- 16047713 TI - Evaluation of the phadiatop test in the diagnosis of allergic sensitization in a general adult population. AB - BACKGROUND: Phadiatop is a commercially available qualitative serological test employed for screening of allergic sensitization in patients with suspected allergic diseases. AIM: The study evaluated the diagnostic accuracy of Phadiatop for the diagnosis of allergic sensitization in a general adult population. METHODS: A total of 469 subjects from the population of A-Estrada (Spain) were selected by age-stratified random sampling (age range, 18-92 years). Phadiatop test (Uni-CAP method) was performed in serum samples from 465 of these subjects. Skin prick tests to a panel of 13 relevant aeroallergens in the studied area (including mites, pollens, moulds, and animal dander) were employed as the reference diagnostic procedure. Subjects with at least a positive skin prick test (> or =4 mm, n= 120) were considered to have allergic sensitization. RESULTS: Phadiatop sensitivity was 70.8% (95% CI 61.7-78.6%), specificity 90.7% (95% CI 87.0-93.5%), positive predictive value 72.6% (95% CI 63.5-80.3%), negative predictive value 89.9% (95% CI 86.2-92.8%), global accuracy 85.6% (95% CI 82.0 88.6%), negative likelihood ratio 0.3 (95% CI 0.2-0.4), and positive likelihood ratio 7.6 (95% CI 5.4-10.8). A high proportion of false-positive Phadiatop cases showed (a) increased total serum IgE levels, (b) significant alcohol consumption, and (c) small-sized (below the diagnostic cut-off) wheal reactions on SPT. A high proportion of false-negative Phadiatop cases showed exclusive storage mite sensitization. Sensitivity and positive predictive value of Phadiatop were somewhat higher among individuals with a history of nasal or bronchial symptoms. CONCLUSIONS: Phadiatop is a valuable tool for the diagnosis of allergic sensitization in a general adult population. However, limitations of the test should be taken into account in similar surveys. PMID- 16047714 TI - Allergenic extracts from Metarhizium canisopliae: obtainment and characterization. AB - Metarhizium anisopliae is used as a biopesticide for insects that damage agricultural plantations like sugar cane and forage plants. In a previous study the sensitization to this fungus of asthmatic patients coming from sugar cane areas was showed. The aims of this work were: to compare crude extracts obtained with Tris-HCl and Coca liquid from several growth phases of M. anisopliae concerning the total content of proteins and their electrophoretic analysis profile; to evaluate in vivo allergic sensitization in Balb/c mice and allergic patients from a sugar cane area, and to characterize the allergenic fractions in the sera of patients positive for the prick test by means of Western-blotting. The extract obtained with Coca liquid on the 16th day was the one that presented the greatest number of proteic fractions, including all those present in the other extracts. Twelve fractions were verified in this extract with approximate molecular weights from 94 to 14 kDa. The allergenicity of the extract obtained on the 16th day was proven by the production of IgE antibodies in Balb/c mice, with titres of 200. Prick tests carried out with the extract of the 16th day in 79 atopic individuals (from sugar cane area), 35 atopic individuals (from urban area) and 11 non- atopic individuals showed respective positivity of 29%, 9% and 0%. The allergenic characterization in vitro was performed by means of Western blotting, and the fractions that reacted with the positive individuals' sera were those of approximate molecular weights of 67 kDa (95%); 20 kDa (55%); 94 kDa (36%); 34 and 36 kDa (23%); 43 and 48 kDa (14%); 16 kDa (9%) and 54kDa (5%). It was concluded that the crude allergenic extract, obtained with Coca liquid from the 16th day growth of Metarhizium anisopliae, contains allergenic fractions and can be used in diagnostic screening tests. PMID- 16047715 TI - Symptoms of the olive pollen allergy: do they really occur only in the pollination season? AB - BACKGROUND: Olive (Olea europaea; O.e.) pollen is a major cause of seasonal respiratory allergy. The O.e. pollination season lasts two months from the beginning of May till the end of June. It was expected that patients with allergic disease from O.e. sensitization were symptomatic only during this period. However, during the last few years, we have observed that the clinical symptoms appear not only during the O.e. pollination season but also during the rest of the year. OBJECTIVE: The aim of this study was to observe and document symptoms of respiratory allergic diseases in the O.e. sensitized patients during the O.e. pollination season and after it. METHODS: One hundred and twenty-seven patients with respiratory allergic disease were enrolled in the study. Allergenic sensitizations were shown by SPT. Finally, patients were split into two groups as monosensitized with O.e. (n=19) and polysensitized (n=108). Patients were assessed by using scores of respiratory allergic disease symptoms and percentage of peak expiratory flow rate values (PEFR %) (only for asthmatic patients) during the O.e. pollination season and after it. RESULTS: Of the patients with O.e. monosensitization, 13 had allergic rhinitis (AR) only while six had allergic asthma (AA) additionally. AR alone and accompanied by AA was present in 84 and 24 polysensitized patients respectively. Eleven patients with O.e. sensitization (57.9 %) and 86 patients with polysensitization (79.6 %) had AR symptoms throughout the year irrespective of the O.e. pollination season. Similarly, three of the O.e. monosensitized and ten of the polysensitized patients with AA had asthmatic symptoms during the O.e. pollination season and also after it. CONCLUSIONS: In the patient group sensitive to O.e. along with other pollen extracts, it was possible to observe symptoms outside the pollination season. However, patients with O.e. monosensitization also had symptoms to a great extent outside the season. PMID- 16047716 TI - Immediate hypersensitivity to quinolones: moxifloxacin cross-reactivity. AB - INTRODUCTION: Immediate hypersensitivity reactions to quinolones are rare. Moxifloxacin is an quinolone chemically different from other fluoroquinolones. We report 6 patients diagnosed with hypersensitivity to different fluoroquinolones in whom the response to moxifloxacin and cross-reactivity with other quinolones was studied. MATERIAL-METHODS: An allergenic study was made by prick and intradermal test with different fluoroquinolones, in all the patients. Single blind oral challenge tests were performed with moxifloxacin in all the patients, with ciprofloxacin in five patients, with levofloxacin in three patients and with ofloxacin in one patient. RESULTS: The skin test performed with moxifloxacin was positive in five patients, and the oral challenge test was positive in all six patients. All the patients had at least one positive skin test with some of the other fluoroquinolones. CONCLUSION: The skin test with different quinolones seems to be sensitive at showing group hypersensitivity, but not at predicting specific tolerance of each drug. We found a high degree of cross-reactivity among fluoroquinolones, so we currently recommend to avoid the group. We did not find that moxifloxacin differed from other fluoroquinolones so we cannot recommend it as a valid therapeutic alternative in patients sensitized to other quinolones. PMID- 16047717 TI - Allergy to freshwater shrimp (Gammarus). AB - We report three new cases of allergy to Gammarus, two of them involving cutaneous symptoms, and the third one with occupational asthma. The results showed exposure to feed containing Gammarus shrimp to be the main cause of the allergic symptoms in the three patients. In all cases the intervention of IgE was demonstrated, with the absence of cross-reactions with other common allergenic arthropods. PMID- 16047718 TI - Not all food additive related reactions originate from commercial foods: chronic urticaria due to home-made canned tomato. AB - Additives and preservatives in commercial foods have been implicated in the etiology of chronic urticaria, but such foods have not been widely accepted. In some countries, as in ours, people prefer to use home-made foodstuffs to avoid potentially hazardous commercial additives. However, not all home-made foodstuffs are safe, especially regarding allergies. In this report, we describe a patient with chronic urticaria due to home-made canned tomato prepared using "tomato drug" as a "safe (!)" additive. PMID- 16047719 TI - Usefulness of montelukast to prevent adverse reactions to COX-2 selective inhibitors: a case report. AB - Nonsteroidal anti-inflammatory drug (NSAID)-sensitivity is a frequent condition in patients with chronic urticaria and/or asthma. The physiopatologic process underlying respiratory and cutaneous reactions probably involves an increased production of cysteinyl leukotrienes. Cyclooxygenase 2 (COX-2) selective inhibitor, has been proposed as the main alternative to control pain and inflammatory diseases in these patients. However, a small percentage of patients with NSAID-induced skin reactions does not even tolerate COX-2 selective inhibitors. We report a very infrequent case of a patient with NSAID, paracetamol and COX-2 selective inhibitors sensitivity in whom we induced tolerance to paracetamol and celecoxib using the leukotriene receptor antagonist montelukast prior to oral challenges. PMID- 16047720 TI - Anaphylaxis caused by cauliflower. AB - BACKGROUND: Cauliflower is a vegetable belonging to the family Cruciferae, genus Brassica, var. Botrytis. METHODS: We report the case of a 70 year-old man who suffered an acute episode consisting of oropharyngeal itching, facial and hand swelling, dyspnea and severe bronchospasm within a few minutes after eating vegetable paella containing cauliflower, green beans, red and green pepper. Due to the severity of the reaction he needed treatment in the emergency room. RESULTS: Skin prick tests with common aeroallergens were positive to Cupressus, Platanus and grass pollen. A strong skin prick test response was obtained with cauliflower and peach lipid transfer protein. Skin prick test with rice, green beans and pepper were negative. Specific IgE determinations were positive to cabbage and cauliflower, and negative to mustard. CONCLUSIONS: The clinical history and the results of the allergologic work-up point out to this patient having experienced an IgE-mediated anaphylactic reaction to cauliflower. To the best of our knowledge, this is the first case report of anaphylaxis due to cauliflower. PMID- 16047721 TI - The disappearing doctor. AB - As medical malpractice premiums continue to rise, physicians are leaving practice in record numbers, and states are taking matters into their own hands. See what the AMA's "crisis states" and, in some cases hospitals, are doing to stem the exodus. PMID- 16047722 TI - Hand in hand: patient and employee satisfaction. AB - When you make your hospital the kind of place where employees can do their best work, you create an environment that attracts patients. PMID- 16047723 TI - Back to basics 2. Communicating with patients and families: leadership strategies. PMID- 16047724 TI - Common ground: hospitals, physicians and outpatient centers. AB - Physician-owned ambulatory surgery centers are projected to keep growing. For at least one hospital, it's a sign to start partnering and stop competing. PMID- 16047725 TI - Joan Soriano: service above self. PMID- 16047726 TI - Communicating hospital distress starts with employees. PMID- 16047727 TI - Physician trustees: maximizing their contribution to governance. PMID- 16047728 TI - Strategic planning. Enhancing patient flow. PMID- 16047729 TI - ICU nurse convicted of murdering six patients. PMID- 16047730 TI - Nursing home 'understaffed' death results: damages awarded. Case on point: Rose Care Inc. v. Ross, 2005 WL 1283679 S.W.3d-AR. PMID- 16047731 TI - AR: RN injured back moving 300 lb. patient: claimant's right to benefits & physician of choice. PMID- 16047732 TI - Male RN alleges sex discrimination & retaliatory termination. Case on point: Rolison v. Bozeman Deaconess Health Services, 111 P.3d 202-MT (2005). PMID- 16047733 TI - An experimental model for study of sialoglycoproteins of human immunodeficiency virus 1 epitope structures. AB - Sialic acid (SA) molecules located terminally on retrovirus glycoproteins (gps) play a key role in virus-cell interactions. The specificity of sialylation of Human immunodeficiency virus 1 (HIV-1) gps has not yet been studied. Looking for a convenient and reproducible experimental virus-cell model for studying the problem mentioned above we compared viral sialoglycoprotein (Sgp) patterns in H9/HTLV III B cells chronically infected with laboratory-adapted HIV-1LAI and MT 2 cells acutely infected with the same virus. Cytosols (CSs) and supernatant concentrates (SNs) from these cells and cell cultures, respectively, following N acetyl-D-[U-14C]-mannosamine ([14C]NAcMan) labeling were subjected to preparative isoelectrofocusing and the obtained fractions were assayed for 14C-incorporation, reverse transcriptase (RT) activity and protein content. Sgp patterns in CSs from the two types of infection were similar. Highly sialylated peaks clustered mainly in the acidic region where the highest 14C-incorporation, RT activity and protein content were found. The 14C-incorporation was higher in CS than in SN. Analysis of CS from MT-2 cells infected with HIV-1 for the markers described above seems to be the experimental approach and model of choice for clinical isolates of HIV 1. PMID- 16047734 TI - Two cis-acting elements in negative RNA strand of Hepatitis C virus involved in synthesis of positive RNA strand in vitro. AB - Sequences at the 3'-ends of both positive and negative strands of Hepatitis C virus (HCV) RNA harbor cis-acting elements required for RNA replication. However, little is known about the properties of the negative RNA strand as a template for the synthesis of positive RNA strand. In this study, a purified recombinant HCV RNA-dependent RNA polymerase (RdRp) was used to investigate the synthesis of positive RNA strand using the 3'-terminal region of negative RNA strand ((-)3'T RNA) as template. A mutagenesis analysis was performed to evaluate the role of the 3'-proximal stem-loop and the first 3'-cytidylate (3'C) of the negative RNA strand in the synthesis of the positive RNA strand. A negative RNA strand of wild type (wt) HCV as template was able to direct the synthesis of a full-length positive RNA strand. Deletion of the 3'-proximal stem-loop resulted in an approximately 90% decrease in RNA synthesis. Disruption of the 3'-proximal stem loop structure by nucleotide substitutions led to a 70-80% decrease in RNA synthesis. However, the restoration of the stem-loop by compensatory mutations in the stem region restored also the RNA synthesis. Likewise, the deletion or substitution of the first 3'C by guanylate (G) led to a 90% decrease in the RNA synthesis; while the substitution by adenylate (A) or uridylate (U) resulted in a 60-80% decrease in the RNA synthesis only. These findings demonstrate that the 3' proximal stem-loop and the first 3'C of the negative RNA strand of HCV are two cis-acting elements involved in the synthesis of the positive RNA strand. PMID- 16047735 TI - The role of ERK1/2 activation in the infection of HeLa cells with Human coxsackievirus B3. AB - Human coxsackievirus B3 (CVB3) is known to trigger in host cells a biphasic activation of extracellular signal-regulated kinase (ERK1/2); i.e., early transient and late sustained activation. In this study, we explored (i) the role of ERK1/2 activation in virus entry into cells and virus replication and (ii) cellular genes influenced by this activation in CVB3-infected HeLa cells. Pretreatment of the cells with an ERK1/2 inhibitor, PD98059 showed that early transient ERK1/2 activation is not be related to virus entry, but late sustained ERK1/2 activation plays a role in virus replication. To identify which cellular genes are influenced by the ERK1/2 activation after virus infection, a cDNA microarray analysis was performed. In HeLa cells pretreated with PD98059 and then infected with the virus, the number of influenced cellular genes was higher compared to that in infected cells not pretreated with the inhibitor (15 vs 77 at 10 mins post infection (p.i.) and 347 vs 91 at 9 hrs p.i. Thus the virus infection affected several host genes through ERK1/2 activation. PMID- 16047736 TI - A correlation between the heterogeneity of hypervariable region 1 of E2 glycoprotein of Hepatitis C virus (HCV) and HCV antibody profile: a case study. AB - A correlation between the heterogeneity of hypervariable region 1 (HVR1) of E2 glycoprotein (gp) and Hepatitis C virus (HCV) antibody profile was investigated. Of 6 patients studied two were in acute phase, two in chronic phase and two showed signs of long-time HCV infection, i.e. liver cirrhosis. All the patients exhibited a vigorous antibody response to viral proteins C, NS3, NS4 and NS5. An antibody response to HVR1 of E2 was found in one patient in acute phase and in one or two patients in chronic phase. Such a response was not found in the two patients with liver cirrhosis. Single-stranded conformation polymorphism (SSCP) and sequence analyses of HVR1 of E2 showed the lowest HVR1 heterogeneity in patients in acute phase and the highest one in those in chronic phase, while the long-time carriers of the virus showed an intermediate heterogeneity. This may reflect a specific interplay between the virus and immune system. The HVR1 heterogeneity may rise in the course of infection as a means of evading the immune pressure. Then, when an organism is unable to clear the virus, because the responses to HVR1 epitopes are weakened or exhausted, a population of less heterogeneous HVR1 variants may be established. PMID- 16047737 TI - Biological and molecular characterization of Indian isolates of Newcastle disease virus from pigeons. AB - Five Newcastle disease virus (NDV) isolates from pigeons were characterized by biological and molecular methods. Four of the five isolates were found to be velogenic with high intracerebral pathogenicity indices (ICPI). The fusion protein cleavage site (FPCS) sequences of these isolates had multiple basic amino acids RRQKRF at positions 112-116 and a phenyl alanine at position 117 characteristic of velogenic isolates. Three of these velogenic isolates were phylogenetically related to mesogenic vaccine virus strain and the fourth one to a few exotic velogenic isolates. The lentogenic isolate obtained in this study was identical with the LaSota strain. PMID- 16047738 TI - The incidence of rotavirus a isolates of G genotype in Thailand in 2002-2004. AB - Rotaviruses are the leading cause of severe gastroenteritis among infants and young children worldwide. Between November 2002 and March 2004, 36 stool specimens of 108 children with acute diarrhea in Bangkok, Thailand were found positive for Rotavirus A (RV-A) by RT-PCR. The 36 isolates were subjected to genotyping by RFLP analysis and direct sequencing of a part of the gene for major outer capsid glycoprotein VP7. The sequences obtained were subjected to phylogenetic analysis. Among the isolates the genotypes G1 (5.6%), G2 (69.4 %) and G9 (25.0 %) were found. Comparison of these results with those of previous studies covering the period of 1982-1999 revealed a changing pattern of RV-A G genotypes and thus contributed to the understanding of RV-A epidemiology in Thailand. Any vaccine to be developed against this virus should target the G9 genotype as one of common global genotypes. PMID- 16047739 TI - Occurrence of Colombian datura virus in Brugmansia hybrids, Physalis peruviana L. and Solanum muricatum Ait. in Hungary. AB - Colombian datura virus (CDV) has been found to infect angel trumpets (Brugmansia spp.) frequently and cape gooseberry (Physalis peruviana) and pepino (Solanum muricatum) sporadically in Hungary. A CDV BRG/H isolate was characterized. It had flexuous thread-like virions of about 750 x 12 nm in size. Host range and symptomathological studies revealed its great similarity to authentic CDV isolates. Nicotiana tabacum cultivars and lines resistant to Potato virus Y (PVYN) either genically or transgenically proved highly susceptible to the BRG/H isolate. Tomato (L. esculentum cvs.) was systemically susceptible to this isolate, but some lines of Lycopersicon hirsutum and L. peruvianum turned out to be resistant. Browallia demissa, Ipomoea purpurea, N. megalosiphon and S. scabrum were demonstrated as new experimental hosts of CDV. The BRG/ H isolate proved to be transmissible by the aphid Myzus persicae Sulz. in a non-persistent manner. Potyvirus-specific coat protein (CP) gene sequences of about 1700 bp from angel trumpet, cape gooseberry and pepino plants were amplified by RT-PCR. The cloned BRG/H CP gene showed a 99.12-99.31% identity with other CDV isolates. CDV has been found for the first time to infect naturally cape gooseberry and pepino. Since the botanical genus name of original hosts of CDV has changed from Datura to Brugmansia, we propose to change the virus name from CDV to Angel trumpet mosaic virus (ATMV). PMID- 16047740 TI - Efficiency of various serological techniques for diagnosing Coxiella burnetii infection. AB - An indirect immunofluorescence assay (IFA) using a recently developed commercial kit for detecting antibodies against Coxiella burnetii (C.b.), the etiological agent of Q fever, has been evaluated using human field serum samples. The IFA was compared with an ELISA and a complement fixation test (CFT). The IFA was based on the corpuscular C.b. phase I and phase II antigens specific to anti-C.b. phase I and II antibodies, respectively. Fifty sera from persons with symptoms of Q fever were examined in this study. The IFA compared with the ELISA showed the sensitivities of 97.7% and 87.2% for IgG and 66.7% and 60.0% for IgM phase II and I antibodies, respectively and the specificities of 100% and 90.0% for IgG and 75.9% and 64.7% for IgM phase II and phase I antibodies, respectively. Due to a limited number of sera positive in the IgA antibody testing, the data presented should be considered with caution. It appears that the IFA strikes a very good balance between high specificity and sensitivity with phase II and phase I IgG antibodies and a less satisfactory one with IgM antibodies. The CFT failed in one of the above aspects showing a good specificity but a poor sensitivity, especially for phase I antibodies. The study demonstrated that the IFA is suitable for diagnosing Q fever and its therapeutic follow-up and is a good candidate for screening sera in large numbers. A certain limitation, especially in testing early stages of the chronic disease, could be a low fluorescence intensity of the IgA positive control in comparison with the IgA negative one. PMID- 16047741 TI - Genotyping of Uruguayan Human adenovirus isolates collected between 1994 and 1998. AB - Adenoviruses are one of the most frequent causative agents of acute lower respiratory infections in infants and young children. Twenty-three adenovirus isolates from nasopharyngeal aspirates of children hospitalized for acute lower respiratory infections in Uruguay between 1994 and 1998 were studied by restriction enzyme analysis. The genomic analysis showed that 60.9% (n = 14) of isolates belonged to the species Human adenovirus C (HAdV-C) and 31.9% (n = 9) to the species Human adenovirus B (HAdV-B). Whereas some isolates could be classified according to the published profiles into genotype or genomic variant, others displayed migration patterns not allowing classification. Eight isolates (89%) of HAdV-B corresponded to the Ad7h genotype that has been associated with severe and fatal pneumonia and necrotizing bronchiolitis in children in South America. The isolates of HAdV-C showed a great variability in accordance with the data published earlier. PMID- 16047742 TI - Expression of S100P gene in cervical carcinoma cells is independent of E7 human papillomavirus oncogene. AB - High-risk human papillomaviruses (HPV) significantly contribute to development of cervical cancer. HPV E7 oncoprotein interferes with the control of cell growth via functional inactivation and/or regulation of multiple molecular targets. Induction of ectopic E7 in breast carcinoma cells has been proposed to decrease transcription of S100P gene, which encodes a calcium-binding protein associated with different types of tumors. We examined a possible relationship between E7 and S100P genes in cervical cell lines. RT-PCR analysis revealed that all HPV positive cell lines expressed approximately equal levels of E7. Out of them, HeLa, CGL3 and SiHa carcinoma cells as well as HCE16/3 immortalized cells expressed also S100P gene. Inhibition of a DNA methylation by 5-aza-2' deoxycytidine (5-aza-dC) in S100P-negative cell lines CGL1 and Caski resulted in induced transcription of S100P, but the normal S100P level in SiHa cells was not further increased. Our results suggest that S100P gene expression is independent of E7 in cervical cell lines and that at least in some cases it is subjected to regulation by DNA methylation. PMID- 16047743 TI - Association of fowl adenovirus serotype 12 with hydropericardium syndrome of poultry in India. AB - Eight fowl adenovirus (FAdV) isolates obtained from different geographical regions of India were typed by a virus-neutralization test (VNT) using rabbit antisera against all the 12 serotypes of FAdV and by PCR for the hyper-variable region of hexon gene combined with restriction fragment length polymorphism (RFLP) analysis using AluI and MboI restriction enzymes. It was found that six isolates belonged to FAdV-4, one to FAdV-12 and one to both of them. This study revealed the involvement of FAdV-12 alone or in association with FAdV-4 in precipitating inclusion body hepatitis--hydropericardium syndrome (IBH-HPS) among poultry flocks in the country. PMID- 16047744 TI - [Aleutian disease in ferrets from New Zealand]. PMID- 16047745 TI - [Campylobacter as a venereal disease in cattle]. AB - Since the introduction of AI, venereal diseases caused by Tritrichomonas fetus and Campylobacter fetus subsp. venerealis haved been eradicated in The Netherlands. Campylobacter fetus subsp. fetus can cause sporadic abortion and early embryonic death. When natural breeding is practised, venereal diseases must be included in the list of differential diagnoses of fertility problems. A case study of a Campylobacter fetus subsp. fetus infection is described. PMID- 16047746 TI - [Become NOW a member of the Interest Association Practice Group]. PMID- 16047747 TI - [FIDIN Antibiotics Report 2004. Organized by the FIDIN Working Group for Antibiotics Legislation. The Hague, the Netherlands, June 2005]. PMID- 16047748 TI - ['Wageningen' makes sure the animal does not get sick, 'Utrecht' cures the animal once it is sick]. PMID- 16047750 TI - [Missed chance or public relations blunder?!]. PMID- 16047749 TI - ['Mammary Gland Health Center Netherlands' starts]. PMID- 16047751 TI - [Beauty is in the eye of the beholder]. PMID- 16047752 TI - [Diagnosis of babesiosis in the dog]. PMID- 16047753 TI - [The VWA increases control over export certification of sheep and goats]. PMID- 16047754 TI - [Is it correct that canned food is better for cats with bladder sediment?]. PMID- 16047755 TI - [Instruments, books and other objects memorable to (almost) forgotten opinions, therapies, buildings, et cetera. The role of Luyendijk]. PMID- 16047756 TI - [Letter to the members of the Dutch Veterinary Medicine Societies Update new quality control management]. PMID- 16047757 TI - [Communication about quality control management]. PMID- 16047758 TI - [GGG supports research to determine the Dutch tick population]. PMID- 16047759 TI - [Differentiation in interests?]. PMID- 16047760 TI - [Veterinary drug use or misuse]. PMID- 16047761 TI - Growing ozone hole "hangover". PMID- 16047762 TI - California looks at tackling indoor air quality. PMID- 16047763 TI - Ridding ships' ballast water of microorganisms. PMID- 16047764 TI - Lessons from endocrine disruption and their application to other issues concerning trace organics in the aquatic environment. AB - In the past 10 years, many thousands of research papers covering the many different aspects of endocrine disruption in the environment have been published. What has been learned from all this research? We have tried to reduce this very large volume of research into a relatively small number of "lessons". Hence, this paper is not a typical review, but instead it summarizes our personal opinions on what we consider are the major messages to have come from all this research. We realize that what has been a lesson to us may have been obvious from the outset to someone more knowledgeable on that particular aspect of the burgeoning field of endocrine disruption. In addition, it is inevitable that others will consider that we have "missed" some lessons that they would have expected to find included in our list. If so, we encourage them to submit them as responses to our paper. Our own lessons range widely, from the design and interpretation of data from fieldwork studies, through some key messages to come out of the very many laboratory studies that have been conducted, to issues around the sources and fates in the environment of endocrine-disrupting chemicals, and finally to the key role of sewage treatment in controlling the concentrations of these chemicals in the aquatic environment. Having (hopefully) learned our lessons, we have then applied them to the difficult issue of how best to approach future concerns about the potential impacts of other new and emerging contaminants (e.g., pharmaceuticals) on wildlife. PMID- 16047765 TI - Conservation of coral reefs through active restoration measures: recent approaches and last decade progress. AB - The scientific discipline of active restoration of denuded coral reef areas has drawn much attention in the past decade as it became evident that this ecosystem does not often recover naturally from anthropogenic stress without manipulation. Essentially, the choices are eitherthe continuous degradation of the reefs or active restoration to encourage reef development. As a result, worldwide restoration operations during the past decade have been recognized as being a major tool for reef rehabilitation. This situation has also stirred discussions and debates on the various restoration measures suggested as management options, supplementary to the traditional conservation acts. The present essay reviews past decade's (1994-2004) approaches and advances in coral reef restoration. While direct coral transplantation is still the primer vehicle of operations used, the concept of in situ and ex situ coral nurseries (the gardening concept), where coral materials (nubbins, branches, spats) are maricultured to a size suitable for transplantation, has been gaining recognition. The use of nubbins (down to the size of a single or few polyps) has been suggested and employed as a unique technique for mass production of coral colonies. Restoration of ship grounding sites and the use of artificial reefs have become common tools for specific restoration needs. Substrate stabilization, 3-D structural consideration of developing colonies, and the use of molecular/biochemical tools are part of novel technology approaches developed in the past decade. Economic considerations for reef restoration have become an important avenue for evaluating success of restoration activities. It has been suggested that landscape restoration and restoration genetics are important issues to be studied. In the future, as coral reef restoration may become the dominant conservation act, there would be the need not only to develop improved protocols but also to define the conceptual bases. PMID- 16047766 TI - Why do organochlorine differences between arctic regions vary among trophic levels? AB - Statistical analysis of organochlorine contaminants (OCs) in marine mammals has shown that, for most OCs, the European Arctic is more contaminated than the Canadian and U.S. Arctic. Recently, comparison of OC concentration ranges in seabirds, arctic cod (Boregadus saida), and zooplankton, found no difference between these regions. To address these inconsistencies, marine food web OC data from the European (central Barents Sea (CBS)) and Canadian Arctic (Northwater Polynya (NOW)) were simultaneously statistically analyzed. In general, concentrations of OCs were greater in seabirds and ringed seals (Phoca hispida) from the CBS as compared to the NOW; consistent with circumpolar trends observed in marine mammals. In contrast, levels of OCs were generally similar in zooplankton and arctic cod between the CBS and NOW. The main exception is HCH which had greater levels in the NOW across all trophic levels because of the greater proximity to sources in eastern Asia. The lack of differences in OC concentrations in zooplankton and Arctic cod from the European and Canadian Arctic suggest that regional differences in OC contamination in the Arctic have evened out. Reduced regional differences were not observed in marine mammals or seabirds because they are long-lived and also acquire contaminants from maternal transfer and hence reflect levels from the past when the European Arctic was more contaminated than the Canadian Arctic. In addition, seabirds may reflect exposure from other areas. This study highlights the potential problem of comparing spatial trends by using means and confidence intervals as compared to simultaneous statistical analysis of raw data. Differences in the spatial trends of OCs between trophic levels in the Arctic are important for consideration when assessing regional differences in spatial and temporal trends of discontinued and current-use contaminants. PMID- 16047767 TI - Nitrous oxide fluxes in three experimental boreal forest reservoirs. AB - Global atmospheric concentrations of nitrous oxide (N2O), a powerful greenhouse gas, continue to increase. While many sources and sinks have been identified, there is little known about how existing and newly constructed reservoirs, such as those created for hydroelectric production, impact current atmospheric N2O concentrations. We hypothesized that N2O fluxes to the atmosphere would increase because enhanced nutrient availability and increased soil respiration following the flooding of soils during reservoir creation would favor denitrification. Furthermore, we hypothesized that emissions would be linked to the amount of organic carbon contained in the flooded landscape. These hypotheses were tested by creating three experimental reservoirs over boreal upland subcatchments that ranged in the amount of organic carbon stored in soils and vegetation. Diffusive surface N2O fluxes within each reservoir were estimated using surface water concentrations of N2O and the thin boundary layer method. Surface fluxes ranged from -1.0 to -3.5 microg N2O m(-2) d(-1), and water column N2O concentrations indicated that contrary to expectations, the reservoirs were acting as slight sinks for atmospheric N2O. This net consumption of N2O was likely related to an excess of labile carbon and low concentrations of oxygen (O2) and nitrate (NO3-) in the flooded soils. Therefore, it is postulated that reservoir creation by flooding boreal soils will likely have little or no net effect of adding additional N2O to the current greenhouse gas (GHG) atmospheric burden, at least over the short term. PMID- 16047768 TI - Historical trends in mercury sedimentation and mixing in the Strait of Georgia, Canada. AB - Seventeen sediment cores collected in the Strait of Georgia reveal a history of mercury contamination that began in the 1860s and include episodic contamination during World War II and in the late 1960s. Surface sediment mercury concentrations ranged from 60 to 420 ng/g dry weight and the current fluxes to sediments are estimated at 5-181 ng cm(-2) a(-1). In one location in Port Moody Arm, a Hg spill seems to have sufficiently poisoned the sediments to eliminate biomixing for about 20 years. Although the surface concentration of Hg is likely to decrease at all stations in coming years, sites in the industrialized Vancouver Harbor and Port Moody Arm will continue to be sufficiently contaminated to endanger benthic organisms. Variations in sedimentation and mixing rates among sites result in surface sediment Hg concentration patterns that do not reflect accurately the distribution of Hg flux. In particular, the concentration of Hg in sediments near the mouth of the Fraser River is low, despite the high Hg load of that river, because of dilution by other particles. A preliminary Hg budget indicates that most of the Hg enters the Strait of Georgia via the Fraser River (2090 kg a(-1)), and that, while burial in Strait of Georgia sediments is a major sink (1800 kg a(-1)), there may be a significant outflow of Hg through Juan de Fuca Strait (approximately 3400 kg a(-1)). PMID- 16047769 TI - Analysis of reduced phosphorus in samples of environmental interest. AB - The combination of ion chromatography (IC) and inductively coupled plasma emission spectroscopy (ICP-ES) was used forthe sensitive and specific detection of hypophosphite (PO2), phosphite (PO3), methylphosphonic acid (MPA), and phosphate (PO4). Application of this technique to a wide range of environmental samples proved that reduced phosphorus was present in some situations including process water from thermal phosphorus plants, drinking water contacting cast iron, and phosphorus corrosion inhibitor used in water treatment and in sewage wastewater. Preliminary testing did not detect high concentrations of reduced phosphorus and phosphine in situations where it was previously reported to be very important, including anaerobic digesters in wastewater treatment plants. The new IC-ICP-ES technique is a promising tool for use in corrosion and soil research where phosphites are likely to be present. PMID- 16047770 TI - Varations in maize pollen emission and deposition in relation to microclimate. AB - The coexistence of genetically modified (GM) crops with conventional crops has become a subject of debate and inquiry. Maize (Zea mays L.) is one of the most cultivated crop plants in the world and there is a need to assess the risks of cross-pollination. Concentration and deposition rate downwind from different sized maize crops were measured during three flowering seasons, together with micrometeorological conditions in the surrounding environment. Pollen release started once the air vapor pressure deficit (VPD) increases above 0.2 to 0.5 kPa. Moreover, the dynamics of release was correlated with the dynamics of VPD surrounding the tassels. Horizontal deposition appeared to follow a power law over short distance downwind from the source, and the dispersal distance increased with the source canopy height and the roughness length of the downwind canopy. This work also provides a data set containing both pollen measurements and contrasting weather conditions to validate dispersal models and further investigate maize pollen dispersal processes. PMID- 16047771 TI - Contribution of dicofol to the current DDT pollution in China. AB - High DDT concentrations and o,p'-DDT/p,p'-DDT ratios observed in the air over Taihu Lake, a lake near Shanghai, China, led us to suggest that current use of dicofol in the area north of the lake was the main source of the measured DDTs. To examine this hypothesis, samples of commercially available formulated dicofol in China were collected in 2003 to measure the impurities of DDT related compounds (DDTs). The o,p'-DDT/p,p'-DDT ratio in the samples was 7.0, close to the observed value in the air over Taihu Lake. Average contents of o,p'-DDT, p,p' CI-DDT, o,p'-DDE, and p,p'-DDT in the samples were 114, 69, 44, and 17 g per kg dicofol, respectively. On the basis of a production and distribution survey, total input of DDTs to the environment from the dicofol use in China was estimated to be 8770 t between 1988 and 2002. "Dicofol type DDT pollution", defined as DDT pollution caused by dicofol use and characterized with high o,p' DDT/p,p'-DDT ratio, might be serious in China, especially in southern and eastern China. The conversion of p,p'-CI-DDT to p,p'-DDE can lead to high p,p'-DDE/p,p' DDT ratio and could mislead the evaluation of p,p'-DDT resident time in the environment. Therefore, more studies on p,p'-CI-DDT in the environment are needed. PMID- 16047772 TI - Sterols and anionic surfactants in urban aerosol: emissions from wastewater treatment plants in relation to background concentrations. AB - Aerosol particles that are emitted from aeration tanks of wastewater treatment plants (WWTPs) can be enriched with environmentally relevant wastewater constituents. In this study, aerosol particles were sampled simultaneously at the pre-aeration tank of a municipal WWTP and at two urban locations approximately 1 km away from the WWTP to evaluate the significance of these aerosol emissions. Moreover, aerosol particles were sampled at a small wastewater irrigation facility and at a rural site. In aerosol particles and wastewater, six sterols (cholesterol, coprostanol, campesterol, beta-sitosterol, stigmasterol, stigmastanol) and anionic surfactants (expressed in terms of methylene blue active substances, MBAS) were quantified. The results showed significantly higher concentrations of sterols and MBAS at the WWTP than atthe urban locations. At the WWTP, average concentrations of cholesterol (848 +/- 321 pg m(-3)), coprostanol (1132 +/- 565 pg m(-3)), and MBAS (132 +/- 43 ng m(-3)) in aerosol were approximately twice as high. This can be attributed to emissions from the treatment tank. Coprostanol, a unique tracer for wastewater, was detected only occasionally at the urban locations. The results of this study show that the aeration of wastewater is a continuously operating local source for organic compounds in aerosol. The wastewater irrigation facility was a minor source of aerosol-bound sterols (coprostanol, 287 +/- 218 pg m(-3)) and anionic surfactants (64 +/- 32 ng m(-3)). Except for coprostanol, all compounds were also present in samples of rural aerosol. PMID- 16047773 TI - Homologue and isomer patterns of polychlorinated dibenzo-p-dioxins and dibenzofurans from phenol precursors: comparison with municipal waste incinerator data. AB - The role of phenol precursors in polychlorinated dibenzo-p-dioxin (PCDD) and dibenzofuran (PCDF) formation in municipal waste incinerators is assessed on the basis of homologue and isomer patterns. Homologue and isomer patterns of PCDD and PCDF congeners formed from phenols both in the gas phase and via particle mediated reactions were studied in an isothermal flow reactor. A mixture of unsubsitituted phenol and 19 chlorinated phenols in relative concentrations found in a municipal waste incinerator (MWI) stack gas was used for this study. PCDD and PCDF homologue and isomer patterns obtained from the phenol experiments were compared with those observed in MWI data. From the phenol experiments, gas-phase formation at 600-700 degrees C favors PCDF formation whereas particle-mediated formation at 400 degrees C favors PCDD formation. Unsubstituted phenol, which was present in high concentration, played a significant role in the formation of PCDD/F congeners under both sets of experimental conditions. PCDD/F distributions in MWI flue gas and fly ash samples were differentfrom those observed in the phenol experiments, suggesting that direct phenol condensation was not the primary route of PCDD/F formation at the incinerators. Gas-phase phenol condensation is a source of dibenzofuran, with subsequent particle-mediated chlorination resulting in PCDF formation. In the case of PCDD formation, phenol condensation may be responsible for the formation of certain highly chlorinated congeners. In this paper we demonstrate the use of homologue and isomer patterns for PCDD/F formation mechanism attribution in municipal waste incinerators. PMID- 16047774 TI - Spatial and temporal variability in air concentrations of short-chain (C10-C13) and medium-chain (C14-C17) chlorinated n-alkanes measured in the U.K. atmosphere. AB - Two studies were carried out on short-chain (C10-C13) and medium-chain (C14-C17) polychlorinated n-alkanes (sPCAs and mPCAs) in U.K. air samples. The first study entailed taking 20 24-h air samples with a pair of Hi-Vol air samplers at the Hazelrigg field station, near Lancaster University. These samples were carefully selected to coincide with times when air masses were predicted to have a fairly constant back trajectory for 24 h and to give a broad spectrum of different origins. The second study was a spatial survey of PCAs in the air at 20 outdoor sites in northern England and four indoor locations in Lancaster, using polyurethane foam (PUF) disk passive air samplers. Levels of the sPCAs in the Hi Vol samples ranged from <185 to 3430 pg m(-3) (average 1130 pg m(-3)) and were higher than those previously measured at this site in 1997. Levels of the mPCAs ranged from <811 to 14500 pg m(-3) (average 3040 pg m(-3)); that is, they were higher than sPCAs. Both sPCA and mPCA air concentrations are of the same order of magnitude as PAH at this site. Back trajectory analysis showed that the history of the air mass in the 48 h prior to sampling had an important effect on the concentrations observed, with overland samples having higher levels than oceanic, implying that the U.K. is probably responsible for most of the PCAs measured in the U.K. atmosphere. Amounts of both sPCAs and mPCAs in the passive air samples followed a rural-urban gradient. PCAs appear to be released from multiple sources around the country, as a result of the diffusive, open industrial and construction use of the technical mixtures. PMID- 16047775 TI - Removal of headspace CO2 increases biological hydrogen production. AB - For biological hydrogen production by fermentation to be a useful method of hydrogen generation, molar yields of hydrogen must be increased. While heat treatment of a soil inoculum increases hydrogen yields by preventing loss of hydrogen to methanogenesis, hydrogen is still lost to acetic acid generation from hydrogen and CO2. To reduce hydrogen losses via acetogenesis, CO2 concentrations in the headspace were substantially reduced during hydrogen production using a chemical scavenger (KOH). CO2 in the headspace was decreased from 24.5% (control) to a maximum of 5.2% during the highest gas production phase, resulting in a hydrogen partial pressure of 87.4%. This reduction in CO2 increased the hydrogen yield by 43% (from 1.4 to 2.0 mol of H2/mol of glucose). The soluble byproducts in all tests consisted primarily of acetate and ethanol. Higher concentrations of ethanol (10.9 mM) remained in solution from bottles with CO2 removal than in the control (1.2 mM), likely as a result of hydrogen inhibition of biological ethanol conversion to acetic acid. These results show that hydrogen production can be increased by removing CO2 in the reactor vessel, likely as a result of suppression of acetogenesis. PMID- 16047776 TI - Bioaccumulation of organochlorines in crows from an indian open waste dumping site: evidence for direct transfer of dioxin-like congeners from the contaminated soil. AB - To assess the significance of waste dumping sites as a source of chemical contamination to ecosystems, we analyzed the residue levels of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), polychlorinated biphenyls (PCBs), and other organochlorines in the breast muscle of crows from a dumping site in the south of Chennai city, South India. Crows from the dumping site contained significantly higher total TEQs (60 +/- 27 pg/g lipid wt) than those from the reference sites (26 +/- 18 pg/g lipid wt). Especially, certain dioxin-like coplanar PCB congeners (Co-PCBs), such as CB-77 and CB-105, whose source is commercial PCBs,were significantly higher in crows from the dumping site than those from the reference sites. Profiles of PCDDs/DFs and Co-PCBs in crows from the dumping site were similar to those of soil at the same site, which was confirmed by principal component analysis. Furthermore, significant positive correlations were obtained between the congener-specific bioconcentration factors (BCFs) of PCDDs/DFs estimated from concentrations in crows and soil from the dumping site and the theoretical BCFs calculated from water-particle and lipid water partitioning coefficients. On the other hand, the estimated BCFs had significant negative correlations with the molecular weight of PCDDs/DFs, indicating that molecular size limits their bioaccumulation. These results suggest that dioxin-like congeners in the soil of the dumping site were transferred directly to the crows through the ingestion of on-site garbage contaminated with soil, rather than through trophic transfer in the ecosystem. The present study provides insight into the ecological impacts of dumping sites. PMID- 16047777 TI - Isotopic signature and impact of car catalysts on the anthropogenic osmium budget. AB - Higher osmium concentrations and lower 187Os/188Os ratios in sediments from urban areas have been linked to anthropogenic osmium sources. Automobile catalytic converters that use platinum group metals (PGM) are a potential source for this Os pollution. We present the first direct Os concentrations and isotopic measurements of catalytic converters for major automobile brands to test the assumption that car catalysts release Os with a distinct signature in the environment. The analysis of four new catalytic converters yields similar low 187Os/188Os ratios (0.1-0.2), suggesting a similar source for the PGM. The Os concentrations measured are in the ppt range (6-228 ppt). From our results, the osmium contribution of the car catalysts to the environment through attrition (wearing and grinding down of the catalyst by friction) is predicted to be low, <0.2 pg Os/m2/year in highly urbanized environment. We show that Os loss from catalysts as volatile OsO4 is important at car catalyst operating temperatures. Moreover, we estimate that car catalysts may be responsible for up to approximately 120 pg Os/m2 deposited per year in urban areas and that part of it may be exported to sedimentary sinks. Car catalytic converters are thus an important anthropogenic osmium source in densely populated areas. The NIST car catalyst standard (SRM-2557, made from recycled used catalysts) yields higher concentrations (up to 721 ppt Os) and a more radiogenic isotopic composition (approximately 0.38), perhaps indicative of Os contamination during its preparation. PMID- 16047778 TI - Extraction of oxidized and reduced forms of uranium from contaminated soils: effects of carbonate concentration and pH. AB - Uranium may present in soil as precipitated, sorbed, complexed, and reduced forms, which impact its mobility and fate in the subsurface soil environment. In this study, a uranium-contaminated soil was extracted with carbonate/ bicarbonate at varying concentrations (0-1 M), pHs, and redox conditions in an attempt to evaluate their effects on the extraction efficiency and selectivity for various forms of uranium in the soil. Results indicate that at least three differentforms of uranium existed in the contaminated soil: uranium(VI) phosphate minerals, reduced U(IV) phases, and U(VI) complexed with soil organic matter. A small fraction of U(VI) could be sorbed onto soil minerals. The mechanism involved in the leaching of U(VI) by carbonates appears to involve three processes which may act concurrently or independently: the dissolution of uranium(VI) phosphate and other mineral phases, the oxidation-complexation of U(IV) under oxic conditions, and the desorption of U(VI)-organic matter complexes at elevated pH conditions. This study suggests that, depending on site-specific geochemical conditions, the presence of small quantities of carbonate/bicarbonate could result in a rapid and greatly increased leaching and the mobilization of U(VI) from the contaminated soil. Even the reduced U(IV) phases (only sparingly soluble in water) are subjected to rapid oxidation and therefore potential leaching into the environment. PMID- 16047779 TI - Isoprene forms secondary organic aerosol through cloud processing: model simulations. AB - Isoprene accounts for more than half of non-methane volatile organics globally. Despite extensive experimentation, homogeneous formation of secondary organic aerosol (SOA) from isoprene remains unproven. Herein, an incloud process is identified in which isoprene produces SOA. Interstitial oxidation of isoprene produces water-soluble aldehydes that react in cloud droplets to form organic acids. Upon cloud evaporation new organic particulate matter is formed. Cloud processing of isoprene contributes at least 1.6 Tg yr(-1) to a global biogenic SOA production of 8-40 Tg yr(-1). We conclude that cloud processing of isoprene is an important contributor to SOA production, altering the global distribution of hygroscopic organic aerosol and cloud condensation nuclei. PMID- 16047780 TI - 1,4-hydroxycarbonyl products of the OH radical initiated reactions of C5-C8 n alkanes in the presence of NO. AB - Alkanes account for approximately 50% of nonmethane organic compounds present in urban atmospheres. Previous studies have shown that hydroxycarbonyls are important products ofthe OH radical initiated reactions of > or = C5 n-alkanes, but isomer-specific identifications and quantifications of these products have not been carried out. In this work, we have used solid-phase microextraction fibers precoated with O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine for on-fiber derivatization of carbonyl-containing compounds with subsequent analyses by combined gas chromatography-mass spectrometry (GC-MS) and GC with flame ionization detection (GC-FID). GC-MS analyses showed the presence of the oximes of 5-hydroxy-2-pentanone and 4-hydroxypentanal from the n-pentane reaction; 5 hydroxy-2-hexanone, 6-hydroxy-3-hexanone, and 4-hydroxyhexanal from the n-hexane reaction; 5-hydroxy-2-heptanone, 6-hydroxy-3-heptanone, 1-hydroxy-4-heptanone, and 4-hydroxyheptanal from the n-heptane reaction; and 5-hydroxy-2-octanone, 6 hydroxy-3-octanone, 7-hydroxy-4-octanone, and 4-hydroxyoctanal from the n-octane reaction. The formation yields of these 1,4-hydroxycarbonyls were determined from GC-FID analyses. By use of the yields of 1,4-hydroxycarbonyls formed from n hexane, n-heptane, and n-octane at 50% relative humidity (and those from n pentane at both 5 and 50% relative humidity), then formation of 1,4 hydroxycarbonyls accounts for 54% of the reaction products from n-pentane, 57% from n-hexane, 51% from n-heptane, and 53% from n-octane. Combined with previously measured yields of carbonyls, alkyl nitrates, and hydroxyalkyl nitrates, we can now accountfor approximately 74-118% of the products formed from the n-pentane through n-octane reactions. PMID- 16047781 TI - Indirect photolysis promoted by natural and engineered wetland water constituents: processes leading to alachlor degradation. AB - Wetland surface waters that received drainage from agricultural fields were probed for constituents that would promote the photodegradation of agriculture herbicides. Alachlor proved to be a good chemical probe for examining indirect photolysis due to its lack of reactivity by either direct photolysis or dark reaction pathways and its ubiquity as an agricultural herbicide. Water samples were taken from natural (Old Woman Creek) and engineered wetlands in Ohio that receive copious amounts of agricultural runoff. Possible photosensitizers including dissolved organic matter (DOM), iron, and nitrate were measured in the samples. In alkaline waters (pH > 7.8), the photochemical degradation of alachlor became important only in the presence of high nitrate levels (approximately equal to 1 mM). In pH-adjusted (approximately 4) samples, the observed degradation rate coefficient increased 3-18 times of that measured at the natural pH. Methanol quenching experiments and kinetics modeling suggest that hydroxyl radical is the principal reactant. The promotion of the reaction at the lower pH was apparently related to the activation of the photochemical pathways associated with the DOM and possibly iron-DOM complexes. The rate coefficients measured for the photodegradation of alachlor in reconstituted DOM isolates (cation-exchanged material with very low iron levels) were similar in magnitude to those measured in natural waters containing low amounts of nitrate and high amounts of DOM. Moreover, these reactions also exhibited a pH dependency. Thus, these results suggest that DOM plays a role in promoting an indirect photolytic mechanism that is highly pH dependent. PMID- 16047782 TI - Cross-coupling of sulfonamide antimicrobial agents with model humic constituents. AB - The oxidative cross-coupling of sulfonamide antimicrobials to constituents of natural organic matter was investigated. Sulfonamide antimicrobials were incubated with surrogate humic constituents in the absence and presence of phenoloxidases (viz., peroxidase, laccase, and tyrosinase) or acid birnessite. Substituted phenols were chosen as simple model constituents to determine the structures in humic substances important for cross-coupling reactions. The extent of sulfonamide transformation was evaluated by the disappearance of the parent compound from solution. Incubation with phenoloxidases in the absence of substituted phenols resulted in little or no sulfonamide transformation. In contrast to this, direct oxidation of sulfonamides by acid birnessite was significant. Inclusion of o-diphenols and 2,6-dimethoxyphenols in reaction mixtures resulted in significant phenoloxidase-mediated transformation of sulfonamides and enhanced antimicrobial transformation in the presence of acid birnessite. Phenolic compounds with other substitution patterns were less effective in promoting sulfonamide transformation. Nuclear magnetic resonance spectroscopy experiments provided direct evidence of peroxidase-mediated covalent cross-coupling of sulfamethazine with syringic and protocatechuic acids. Our results indicate that sulfonamide antimicrobials may be chemically incorporated into humic substances. This may result in their diminished mobility, bioavailability, and biological activity. PMID- 16047783 TI - Oxidative transformation of fluoroquinolone antibacterial agents and structurally related amines by manganese oxide. AB - Various members of the popular fluoroquinolone antibacterial agents (FQs) have been frequently detected in municipal wastewater and surface water bodies in recent years. This study was conducted to gain a better understanding of the fate of FQs in the sediment-water environment. Seven FQs were examined for adsorptive and oxidative interactions with delta-MnO2 under environmental conditions and exhibited reactivity in the order of ciprofloxacin approximately enrofloxacin approximately norfloxacin approximately ofloxacin > lomefloxacin > pipemidic acid >> flumequine. Four amines that are structurally related to the aniline and piperazine functional groups of FQs showed reactivity to oxidation by delta-MnO2 in the order of 1-phenylpiperazine > aniline > N-phenylmorpholine > 4 phenylpiperidine. Comparison among the above compounds clearly indicates thatthe piperazine moiety of FQs is the predominant adsorptive and oxidative site to MnO2. Product analyses showed that oxidation by MnO2 results in dealkylation and hydroxylation at the piperazine moiety of FQs, with the quinolone ring essentially intact. The reaction kinetics, reactivity comparison, and product characterization point to a surface reaction mechanism that likely begins with formation of a surface complex between FQ and the surface-bound MnIV, followed by oxidation at the aromatic N1 atom of FQ's piperazine moietyto generate an anilinyl radical intermediate. The radical intermediates subsequently undergo N dealkylation, C-hydroxylation, and possibly coupling to yield a range of products. Even though the quinolone ring appears to be stable with respect to MnO2, it affects the overall reactivity and potentially product distribution of FQs via substituent effects. Results of this study strongly suggest that manganese oxides commonly present in soils will likely play an important role in the abiotic degradation of fluoroquinolone antibacterial agents in the environment. PMID- 16047784 TI - Seasonal temperature fluctuations induces rapid inactivation of Cryptosporidium parvum. AB - This study measured the inactivation rate of bovine genotype A Cryptosporidium parvum oocysts attributable to diurnal oscillations of ambient temperature and solar radiation typical of California rangelands and dairies from spring through autumn. We first measured the relationship between air temperature and the internal temperature of bovine feces exposed to sunlight on commercial operations throughout California. Once maximum air temperature exceeded the mid 20 degrees C, diurnal thermal regimes of bovine fecal material exhibited peaks of over 40, 50, 60, and 70 degrees C. These diurnal thermal regimes were emulated using a thermocycler, with oocysts suspended in distilled water or fecal-water mix. Using oral inoculations of 10(5) C. parvum oocysts per neonatal Balb/c mouse (>1000 fold the ID50), no infections were observed using 1 to 5-day cycles of these thermal regimes. Loss of infectivity induced bythese thermal regimes was primarily due to partial or complete in vitro excystation during the first 24-h diurnal cycle and secondarily to thermal inactivation of the remaining intact or partial oocysts. These results suggest that as ambient conditions generate internal fecal temperatures > or = 40 degrees C via conduction, radiation, and convection, rapid environmental inactivation occurs at a rate of > or = 3.27 log reduction d(-1) for C. parvum oocysts deposited in the feces of cattle. PMID- 16047785 TI - Speciation of Zn associated with diatoms using X-ray absorption spectroscopy. AB - The long- and short-term interactions between zinc, an essential but also toxic element, and freshwater and marine diatoms are not well understood partly because of a lack of information on Zn speciation on the surface and inside the cells. In this work, interactions of aqueous Zn2+ with marine (Skeletonema costatum) and freshwater (Achnanthidium minutissimum, Navicula minima, and Melosira varians) diatoms were studied using conventional macroscopic techniques, while the local atomic structure of metal ions adsorbed on their surface or incorporated into the cells was characterized by in-situ Zn K-edge X-ray absorption fine structure (XAFS) spectroscopy on both intact and liophylized samples. At the cell surface for all diatom species studied, Zn was tetrahedrally coordinated with oxygen at approximately 2.00 +/- 0.02 A and monodentately bonded to one or two carboxylate groups; these results are consistent with the surface speciation model developed from macroscopic adsorption experiments. The atomic environment of Zn incorporated into freshwater diatoms during long-term growth in normal nutrient media was distinctly differentfrom that of adsorbed Zn: it was dominated by O (and/or N) neighbors in a tetrahedral arrangement at 1.97 +/- 0.02 A in the first atomic shell, with the presence of 1 phosphorus and 2 carbons in the Zn second shell. Contrasting speciation of intracellular zinc was revealed for the marine species Skeletonema costatum in which Zn was coordinated to 2 O/N atoms and 2 sulfur groups in the form of cysteine-histidine complexes and/or zinc thiolate clusters. These new structural data strongly suggest: (i) the predominant > R-COO ligand binding of Zn atthe diatom surface; (ii) the nonspecific storage of Zn in the form of carboxylate/phosphate groups inside the cell of freshwater species; and (iii) the highly specific thiol-ligand coordination of intracellular zinc for marine S. costatum species. PMID- 16047786 TI - Cr(VI) reduction and immobilization by magnetite under alkaline pH conditions: the role of passivation. AB - This study investigated Cr(VI) reduction and immobilization by magnetite under alkaline pH conditions similar to those present at the Hanford site. Compared to acidic and neutral pH, chromium(VI) reduction by magnetite at high pH conditions is limited (<20% of potential reduction capacity), and the extent of reduction does not vary significantly with increasing NaOH concentration. This is due to the formation of maghemite, goethite, and/or Fe1-xCrxOOH, which may form a passivation layer on the magnetite surface, stopping further chromate reduction. Maghemite is formed in lower NaOH concentrations. The extent of goethite formation increases with NaOH concentration. Goethite may be formed through two mechanisms: (i) dissolution of magnetite leads to the precipitation of goethite and/or (ii) dissolution of newly formed maghemite intermediate, followed by precipitation of goethite. Extended X-ray absorption fine structure spectroscopy shows that Cr has a similar structural environment at alkaline pH as at acidic and circumneutral conditions. PMID- 16047787 TI - Cesium desorption from illite as affected by exudates from rhizosphere bacteria. AB - Biogeochemical processes in the rhizosphere can significantly alter interactions between contaminants and soil minerals. In this study, several strains of bacteria that exude aluminum (Al)-chelating compounds were isolated from the rhizosphere of crested wheatgrass (Agropyron desertorum) collected from the Idaho National Laboratory (INL). We examined the effects of exudates from bacteria in the genera Bacillus, Ralstonia, and Enterobacter on cesium (Cs) desorption from illite. Exudates from these strains of bacteria significantly enhanced Cs desorption from illite. In addition, Cs desorption increased with increasing Bacillus exudate concentrations. Cesium desorption from illite as a function of both exudate type and concentration was positively correlated with Al dissolution, suggesting that the Al-complexing ability of the exudates played an important role in enhancing Cs desorption. The density of frayed edge sites (FES) on illite increased as a result of treatment with bacterial exudates, while the Cs/K selectivity of FES decreased. These results suggestthat exudates from bacteria isolated from the rhizosphere can enhance Cs desorption from frayed edges of illite and, therefore, can alter Cs availability in micaceous soils. PMID- 16047788 TI - Modeling the influence of intermittent rain events on long-term fate and transport of organic air pollutants. AB - The deposition of particles and substances in air is under strong influence of the precipitation patterns of the atmosphere. Most multimedia models, like type III Mackay models, treat rain as a continuous phenomenon. This may cause severe overestimation of the substance removal from the atmosphere through wet deposition and an underestimation of travel distances, leading to the following questions: How strong is the influence of the intermittent character of rain on concentrations, residence times, deposited fractions, and characteristic transport distances of different substances in air? Is there an expression that can provide an accurate approximation to be used in steady-state multimedia models? Assuming a periodically intermittent rain, the mass of an emitted substance that is present in the air compartment is calculated as a function of the deposition rate constants during dry and wet periods and the durations of these periods. In this paper, results for 300 different organic chemicals are presented and illustrated in more detail for four typical substances, showing the following: (i) Deposition velocities can be up to 4 orders of magnitude higher during rain events than during dry periods, especially for persistent substances with low Henry constant. (ii) For substances with a short reaction time (residence time as determined by atmospheric degradation alone) (e.g., propoxur), the assumption of continuous rain may lead to an underestimation of the atmospheric residence time and travel distance by up to 3 orders of magnitude. For this group of substances, the residence time during dry period provides a good estimate of the overall atmospheric residence time. (iii) For substances with reaction times close to the duration of the dry period, the behavior is driven by the length of the time interval between two rain events, as for example, for methomyl. (iv) For very persistent substances such as pentachloronitrobenzene or carbon tetrachloride, the continuous rain approximation provides a good estimate. On the basis of these findings, an accurate but simple approximation is provided by eq 17 for the incorporation of intermittent rain behavior in steady-state multimedia models. PMID- 16047789 TI - Bioaccumulation factors for PCBs revisited. AB - Bioaccumulation factors (BAFs) for individual polychlorinated biphenyl (PCB) congeners in Barents Sea and White Sea marine calanoid copepods were 1-3 orders of magnitude higher than BAFs in the same species in Canadian and Alaskan Arctic Ocean areas, and in freshwater plankton (Lake Ontario) reported from the mid- to early 1980s. The present study reviews variability in PCB BAFs from the North American Great Lakes and the Arctic Ocean, and discusses possible explanations for the large variation among different studies. BAFs are higher in recent arctic marine and Great Lakes studies than previously reported, and they are at least 10 times higher than those predicted from the octanol-water partition coefficient (KOW). If the recent high BAFs are realistic, it means that earlier reported BAFs are too low. This is likely due to earlier erroneously high quantification of water PCB concentrations, and it implies that bioaccumulation in zooplankton is more efficient than previously assumed. Evidence is presented supporting that also trophic transfer and biomagnification of PCBs in zooplankton leads to BAFs well above those predicted by simple equilibrium partitioning. Overall, miss measurement of water PCB concentrations and biomagnification contribute significantly to variability in BAFs for PCBs within and among studies. This large variability of BAFs for PCBs in zooplankton illustrated in the present study is of importance for future assessments of potential new bioaccumulative chemicals that rely on measured BAFs, such as the European Union Registration, Evaluation and Authorization of Chemicals program (REACH). PMID- 16047790 TI - Organic coprecipitates with calcite: NMR spectroscopic evidence. AB - Dissolved organic ligands are well known to interact strongly with the calcite surface, altering precipitation and dissolution rates, crystal morphology, and possibly the ability of calcite to sequester metal contaminants. We show, using NMR spectroscopic techniques, that some of the citrate molecules present in a solution of precipitating calcite are incorporated structurally into the calcite crystal. Calcite grown by a seeded constant-addition method contains approximately 1 wt % coprecipitated citrate and yields 13C{1H} cross-polarization magic-angle spinning NMR spectra that contain broad peaks for citrate plus a signal from carbonate. Results from 13C{1H} heteronuclear correlation NMR experiments show that citrate is located in close spatial proximity to carbonate groups. In addition, calcite/citrate coprecipitates contain about 0.4 wt % excess water, which is not present as fluid inclusions, and some of which occurs as rigid structural water. These results suggestthat water and hydrogen-bonding interactions play a role in the interface between included organic molecules and the calcite host. Additional NMR data obtained for calcite coprecipitates of aspartic and glutamic acids suggest they are also incorporated structurally but at concentrations much lower than for citrate, whereas no evidence was found for phthalate incorporation. PMID- 16047791 TI - Oxidative microbial degradation of 2,4,6-trinitrotoluene via 3-methyl-4,6 dinitrocatechol. AB - A novel pathway for biodegradation of 2,4,6-trinitrotoluene (TNT) was investigated where TNT was the sole carbon, nitrogen, and energy source. Results showed the ability of microorganismsto metabolize TNT through removal of a nitro group, oxygenation of the aromatic ring, and production of a metabolite that is typically a precursor to oxygenolytic ring cleavage. Nitrite production was observed in active systems, and TNT degradation activity was repeatable and transferable. The metabolic intermediate, 3-methyl-4,6-dinitrocatechol, was positively identified through stable isotope mass spectrometry and tandem mass spectrometry. Experimentation with 14C-TNT showed >3% 14C-labeled CO2 in active systems after 30 d exposure to microorganisms. An increasing fraction of 14C labeled material was associated with biomass with time, where 11.41 +/- 2.91% and 17.09 +/- 1.49% of 14C was associated with biomass in active systems after 20 and 30 d, respectively, as compared with 5.68 +/- 1.33% and 6.08 +/- 1.27% in inactive systems. Parallel degradation of TNT and production of organic metabolites and nitrite were observed in shake flasks constructed with soil from historically contaminated sites, indicating that the novel pathway identified herein is disturbed in the environment. Therefore, results presented provide evidence of a previously unreported pathway for oxidative degradation of TNT. PMID- 16047792 TI - Effect of vapor source-building separation and building construction on soil vapor intrusion as studied with a three-dimensional numerical model. AB - A three-dimensional numerical model of the soil vapor-to-indoor air pathway is developed and used as a tool to anticipate not-yet-measured relationships between the vapor attenuation coefficient, alpha (indoor air concentration/source vapor concentration), and vapor source-building lateral separation, vapor source depth, and building construction characteristics (depth of building foundation) for nondegrading chemicals. The numerical model allows for diffusive and advective transport, multicomponent systems and reactions, spatially distributed foundation cracks, and transient indoor and ambient pressure fluctuations. Simulations involving different lateral separations between the vapor source and building show decreasing alpha values with increasing lateral separation. For example, alpha is 2 orders of magnitude less when a 30 m x 30 m source located 8 m below ground surface is displaced from the edge of the building by 20 m. The decrease in alpha with increasing lateral separation is greater for shallower source depths. For example, alpha is approximately 5 orders of magnitude less when a 30 m x 30 m source located 3 m below ground surface is displaced from the edge of the building by 20 m. To help visualize the effects of changing vapor source building separations, normalized vapor concentration contour plots for both horizontal and vertical cross sections are presented for a sequence of lateral separations ranging from the case in which the 30 m x 30 m source and 10 m x 10 m building footprint centers are collocated to shifting of the source positioning by 50 m. Simulations involving basement and slab-on-grade constructions produce similar trends. In addition, when buildings are overpressurized to create outflow to soil gas on the order of 1-3 L/min, emissions to indoor air are reduced by over 5 orders of magnitude relative to intrusion rates at zero building underpressurization. The results are specific to simulations involving homogeneous soil properties, nondegrading chemicals, steady source concentrations and building underpressurizations, and the geometries studied in this work. PMID- 16047793 TI - Modeling kinetics of Cu and Zn release from soils. AB - Kinetics of Cu and Zn release from soil particles was studied using two surface soils with a stirred-flow method. Different solution pH, dissolved organic matter (DOM) concentrations, and flow rates were tested in this study. A model for kinetics controlled sorption/desorption reactions between soils and solutions was globally fit to all experimental data simultaneously. Results were compared to a model that assumes local instantaneous equilibrium. We obtained one unique set of model parameters applicable to different pH, dissolved organic carbon (DOC), and flow conditions. We included DOM complexation of copper ions, which decreased their sorption. The effect of pH was included by assuming proton competition with metal ions for binding sites on soil particles. These results provide the basis for developing predictive models for metal release from soil particles to surface waters and soil solution. PMID- 16047794 TI - Determination of single particle mass spectral signatures from light-duty vehicle emissions. AB - In this study, 28 light-duty gasoline vehicles (LDV) were operated on a chassis dynamometer at the California Air Resources Board Haagen-Smit Facility in El Monte, CA. The mass spectra of individual particles emitted from these vehicles were measured using aerosol time-of-flight mass spectrometry (ATOFMS). A primary goal of this study involves determining representative size-resolved single particle mass spectral signatures that can be used in future ambient particulate matter source apportionment studies. Different cycles were used to simulate urban driving conditions including the federal testing procedure (FTP), unified cycle (UC), and the correction cycle (CC). The vehicles were selected to span a range of catalytic converter (three-way, oxidation, and no catalysts) and engine technologies (vehicles models from 1953 to 2003). Exhaust particles were sampled directly from a dilution and residence chamber system using particle sizing instruments and an ATOFMS equipped with an aerodynamic lens (UF-ATOFMS) analyzing particles between 50 and 300 nm. On the basis of chemical composition, 10 unique chemical types describe the majority of the particles with distinct size and temporal characteristics. In the ultrafine size range (between 50 and 100 nm), three elemental carbon (EC) particle types dominated, all showing distinct EC signatures combined with Ca, phosphate, sulfate, and a lower abundance of organic carbon (OC). The relative fraction of EC particle types decreased as particle size increased with OC particles becoming more prevalent above 100 nm. Depending on the vehicle and cycle, several distinct OC particle types produced distinct ion patterns, including substituted aromatic compounds and polycyclic aromatic hydrocarbons (PAH), coupled with other chemical species including ammonium, EC, nitrate, sulfate, phosphate, V, and Ca. The most likely source of the Ca and phosphate in the particles is attributed to the lubricating oil. Significant variability was observed in the chemical composition of particles emitted within the different car categories as well as for the same car operating under different driving conditions. Two-minute temporal resolution measurements provide information on the chemical classes as they evolved during the FTP cycle. The first two minutes of the cold start produced more than 5 times the number of particles than any other portion of the cycle, with one class of ultrafine particles (EC coupled with Ca, OC, and phosphate) preferentially produced. By number, the three EC with Ca classes (which also contained OC, phosphate, and sulfate) were the most abundant classes produced by the nonsmoking vehicles. The smoker category produced the highest number of particles, with the dominant classes being OC comprised of substituted monoaromatic compounds and PAHs, coupled with Ca and phosphate, thus suggesting used lubricating oil was associated with many of these particles. These studies show, by number, EC particles dominate gasoline emissions in the ultrafine size range particularlyforthe lowest emitting newer vehicles, suggesting the EC signature alone cannot be used as a unique tracer for diesels. This represents the first report of high time- and size-resolved chemical composition data showing the mixing state of nonrefractory elements in particles such as EC for vehicle emissions during dynamometer source testing. PMID- 16047795 TI - Inter-comparison of laser photoacoustic spectroscopy and gas chromatography techniques for measurements of ethene in the atmosphere. AB - Laser photoacoustic spectroscopy (LPAS) is highly suitable for the detection of ethene in air due to the overlap between its strongest absorption lines and the wavelengths accessible by high-powered CO2 lasers. Here, we test the ability of LPAS to measure ethene in ambient air by comparing the measurements in urban air with those from a gas chromatography flame-ionization detection (GC-FID) instrument. Over the course of several days, we obtained quantitative agreement between the two measurements. Over this period, the LPAS instrument had a positive offset of 330 +/- 140 pptv (parts-per-trillion by volume) relative to the GC-FID instrument, possibly caused by interference from other species. The detection limit of the LPAS instrument is currently estimated around 1 ppbv and is limited by this offset and the statistical noise in the data. We conclude that LPAS has the potential to provide fast-response measurements of ethene in the atmosphere, with significant advantages over existing techniques when measuring from moving platforms and in the vicinity of emission sources. PMID- 16047796 TI - Trace analysis of bromate, chlorate, iodate, and perchlorate in natural and bottled waters. AB - A simple and rapid method has been developed to simultaneously measure sub microg/L quantities of the oxyhalide anions bromate, chlorate, iodate, and perchlorate in water samples. Water samples (10 mL) are passed through barium and hydronium cartridges to remove sulfate and carbonate, respectively. The method utilizes the direct injection of 10 microL volumes of water samples into a liquid chromatography-tandem triple-quadrupole mass spectrometry (LC-MS/MS) system. Ionization is accomplished using electrospray ionization in negative mode. The method detection limits were 0.021 microg/L for perchlorate, 0.045 microg/L for bromate, 0.070 microg/L for iodate, and 0.045 microg/L for chlorate anions in water. The LC-MS/MS method described here was compared to established EPA methods 300.1 and 317.1 for bromate analysis and EPA method 314.0 for perchlorate analysis. Samples collected from sites with known contamination were split and sent to certified laboratories utilizing EPA methods for bromate and perchlorate analysis. At concentrations above the reporting limits for EPA methods, the method described here was always within 20% of the established methods, and generally within 10%. Twenty-one commercially available bottled waters were analyzed for oxyhalides. The majority of bottled waters contained detectable levels of oxyhalides, with perchlorate < or = 0.74 microg/L, bromate < or = 76 microg/L, iodate < or = 25 microg/ L, and chlorate < or = 5.8 microg/L. Perchlorate, iodate, and chlorate were detectable in nearly all natural waters tested, while bromate was only detected in treated waters. Perchlorate was found in several rivers and reservoirs where itwas not found previously using EPA 314.0 (reporting limit of 4 microg/L). This method was also applied to common detergents used for cleaning laboratory glassware and equipmentto evaluate the potential for sample contamination. Only chlorate appeared as a major oxyhalide in the detergents evaluated, with concentrations up to 517 microg/g. Drinking water treatment plants were also evaluated using this method. Significant formations of chlorate and bromate are demonstrated from hypochlorite generation and ozonation. From the limited data set provided here, it appears that perchlorate is a ubiquitous contaminant of natural waters at trace levels. PMID- 16047797 TI - Biomonitoring of mercury exposure with single human hair strand. AB - Hair samples continue to be used extensively for biomonitoring of mercury (Hg) exposure. Routine methods require a bundle of 100-150 hair strands and involve chemical digestion. Recently, Hg analyzers that combine combustion, gold amalgamation, and atomic absorption spectrometry (C-GA-AAS) became commercially available. This method was shown to provide quick and sensitive measurements in solid samples such as hair. The objective of this study was to validate C-GA-AAS for measuring total Hg in single hair strands as an alternative method for Hg biomonitoring. Hair samples from 12 women with a wide range of Hg exposure were obtained from two projects conducted in Brazil and Canada. A 1:1 relationship was observed between C-GA-AAS and the established cold vapor atomic absorption spectrometry (CV-AAS) for analysis of 1-cm segments from a bundle of hair. For individual hair variability, the average relative standard deviation (RSD) of Hg between hair strands was 6.5 +/- 2.8%, thus justifying the use of single hair strand for biomonitoring. With a limit of quantification of 0.10 ng of total Hg, a single hair strand can be used to assess monthly exposure. This technique will facilitate routine biomonitoring and thus help prevent Hg poisoning among the public. PMID- 16047798 TI - Chromium-removal processes during groundwater remediation by a zerovalent iron permeable reactive barrier. AB - Solid-phase associations of chromium were examined in core materials collected from a full-scale, zerovalent iron permeable reactive barrier (PRB) at the U.S. Coast Guard Support Center located near Elizabeth City, NC. The PRB was installed in 1996 to treat groundwater contaminated with hexavalent chromium. After eight years of operation, the PRB remains effective at reducing concentrations of Cr from average values >1500 microg L(-1) in groundwater hydraulically upgradient of the PRB to values <1 microg L(-1) in groundwater within and hydraulically downgradient of the PRB. Chromium removal from groundwater occurs at the leading edge of the PRB and also within the aquifer immediately upgradient of the PRB. These regions also witness the greatest amount of secondary mineral formation due to steep geochemical gradients that result from the corrosion of zerovalent iron. X-ray absorption near-edge structure (XANES) spectroscopy indicated that chromium is predominantly in the trivalent oxidation state, confirming that reductive processes are responsible for Cr sequestration. XANES spectra and microscopy results suggest that Cr is, in part, associated with iron sulfide grains formed as a consequence of microbially mediated sulfate reduction in and around the PRB. Results of this study provide evidence that secondary iron-bearing mineral products may enhance the capacity of zerovalent iron systems to remediate Cr in groundwater, either through redox reactions at the mineral-water interface or by the release of Fe(II) to solution via mineral dissolution and/or metal corrosion. PMID- 16047799 TI - Removal of heavy metals from mine waters by natural zeolites. AB - In this study, we investigated the removal of Fe, Pb, Cd, and Zn from synthetic mine waters by a natural zeolite. The emphasis was given to the zeolite's behavior toward a few cations in competition with each other. Pb was removed efficiently from neutral as well as from acidic solutions, whereas the uptake of Zn and Cd decreased with low pH and high iron concentrations. With increasing Ca concentrations in solution, elimination of Zn and Cd became poorer while removal of Pb remained virtually unchanged. The zeolite was stable in acidic solutions. Disintegration was only observed below pH 2.0. Forward- and back-titration of synthetic acidic mine water were carried out in the presence and absence of zeolite to simulate the effects of a pH increase by addition of neutralizing agents and a re-acidification which can be caused by subsequent mixing with acidic water. The pH increase during neutralization causes precipitation of hydrous ferric oxides and decreased dissolved metal concentrations. Zeolite addition further diminished Pb concentrations but did not have an effect on Zn and Cd concentrations in solution. During re-acidification of the solution, remobilization of Pb was weaker in the presence than in the absence of zeolite. No substantial differences were observed for Fe, Cd, and Zn immobilization. The immobilization of the metals during pH increase and the subsequent remobilization caused by re-acidification can be well described by a geochemical equilibrium speciation model that accounts for metal complexation at hydrous ferric oxides, for ion exchange on the zeolite surfaces, as well as for dissolution and precipitation processes. PMID- 16047800 TI - Development of an E-H2O2/TiO2 photoelectrocatalytic oxidation system for water and wastewater treatment. AB - In this study, an innovative E-H2O2/TiO2 (E-H2O2 = electrogenerated hydrogen peroxide) photoelectrocatalytic (PEC) oxidation system was successfully developed for water and wastewater treatment. A TiO2/Ti mesh electrode was applied in this photoreactor as the anode to conduct PEC oxidation, and a reticulated vitreous carbon (RVC) electrode was used as the cathode to electrogenerate hydrogen peroxide simultaneously. The TiO2/Ti mesh electrode was prepared with a modified anodic oxidation process in a quadrielectrolyte (H2SO4-H3PO4-H2O2-HF) solution. The crystal structure, surface morphology, and film thickness of the TiO2/Ti mesh electrode were characterized by X-ray diffraction and scanning electron microscopy. The analytical results showed that a honeycomb-type anatase film with a thickness of 5 microm was formed. Photocatalytic oxidation (PC) and PEC oxidation of 2,4,6-trichlorophenol (TCP) in an aqueous solution were performed under various experimental conditions. Experimental results showed that the TiO2/Ti electrode, anodized in the H2SO4-H3PO4-H2O2-HF solution, had higher photocatalytic activity than the TiO2/Ti electrode anodized in the H2SO4 solution. It was found that the maximum applied potential would be around 2.5 V, corresponding to an optimum applied current density of 50 microA cm(-2) under UV A illumination. The experiments confirmed that the E-H2O2 on the RVC electrode can significantly enhance the PEC oxidation of TCP in aqueous solution. The rate of TCP degradation in such an E-H2O2-assisted TiO2 PEC reaction was 5.0 times that of the TiO2 PC reaction and 2.3 times that of the TiO2 PEC reaction. The variation of pH during the E-H2O2-assisted TiO2 PEC reaction, affected by individual reactions, was also investigated. It was found that pH was well maintained during the TCP degradation in such an E-H2O2/TiO2 reaction system. This is beneficial to TCP degradation in an aqueous solution. PMID- 16047801 TI - Recovery and fractionation of phosphorus retained by lightweight expanded shale and masonry sand used as media in subsurface flow treatment wetlands. AB - Most subsurface flow treatment wetlands, also known as reed bed or root zone systems, use sand or gravel substrates to reduce organics, solids, and nutrients in septic tank effluents. Phosphorus (P) retention in these systems is highly variable and few studies have identified the fate of retained P. In this study, two substrates, expanded shale and masonry sand, were used as filter media in five subsurface flow pilot-scale wetlands (2.7 m3). After 1 year of operation, we estimated the annual rate of P sorption by taking the difference between total P (TP) of substrate in the pilot cells and TP of substrate not exposed to wastewater (control). Means and standard deviations of TP retained by expanded shale were 349 +/- 171 mg kg(-1), respectively. For a substrate depth of 0.9 m, aerial P retention by shale was 201 +/- 98.6 g of P m(-2) year(-1), respectively. Masonry sand retained an insignificant quantity of wastewater P (11.9 +/- 21.8 mg kg(-1)) and on occasion exported P. Substrate samples were also sequentially fractionated into labile P, microbial P, (Fe + Al) P, humic P, (Ca + Mg) P, and residual P. In expanded shale samples, the greatest increase in P was in the relatively permanent form of (Fe + Al) P (108 mg kg(-1)), followed by labile P (46.7 mg kg(-1)) and humic P (39.8 mg kg(-1)). In masonry sand, there was an increase in labile P (9.71 mg kg(-1)). Results suggest that sand is a poor candidate for long-term P storage, but its efficiency is similar to that reported for many sand, gravel, and rock systems. By contrast, expanded shale and similar products with high hydraulic conductivity and P sorption capacity could greatly improve performance of P retention in constructed wetlands. PMID- 16047802 TI - Adsorption of fluoride on zirconium(IV)-impregnated collagen fiber. AB - A novel adsorbent, zirconium(IV)-impregnated collagen fiber, was prepared. Zr(IV) was uniformly dispersed in collagen fiber, mainly through chemical bonds, and was able to withstand the extraction of water. This adsorbent is effective for the removal of fluoride from aqueous solutions. The adsorption capacity was 2.29 mmol/g at pH = 5.5 when 5.00 mmol/L fluoride solution was adsorbed by use of 0.100 g of adsorbent, and the extent of removal was 97.4% when the adsorbent dose was 0.300 g. The adsorption isotherms were well fitted by the Langmuir equation, and the maximum adsorption capacities calculated by the Langmuir equation were close to those determined by experiment. The adsorption capacity increased with rising temperature. These facts imply that the mechanism of chemical adsorption might be involved in the adsorption process of fluoride on the absorbent and that fluorides are adsorbed in the form of monolayer coverage on the surface of the adsorbent. The adsorption kinetics of fluoride onto Zr(IV)-impregnated collagen fiber could be described by Lagergren's pseudo-first-order rate mode. The investigation on desorption indicated that this adsorbent is easily regenerated by use of dilute NaOH solution. PMID- 16047803 TI - Inactivation of Microcystis aeruginosa by continuous electrochemical cycling process in tube using Ti/RuO2 electrodes. AB - Algae in waters often bring about influence in drinking water supplies. In this study, an electrochemical tube employing titanium coated with RuO2 as anode was constructed for inactivation of cyanobacteria (often called bluegreen algae) Microcystis aeruginosa. Suspensions containing M. aeruginosa (2-4 x 10(9) L(-1)) were exposed to current densities ranging from 1 to 10 mA cm(-2) in a detention time of 52 min. The variations of cell density, chlorophyll-a, optical density, pH, and conductivity were examined during the treatment. After 3.5 min the population of M. aeruginosa dropped rapidly and was reduced from 3 x 10(9) to 0.6 x 10(9) L(-1) after 52 min at current densities from 5 to 10 mA cm(-2). The cell density and optical density of M. aeruginosa decreased proportionally to the current density and the detention time. Scanning electron microscopy investigation of algae revealed surface damage and apparent leakage of intracellular contents after electrochemical cycling process. Due to the damage of cells, the chlorophyll-a released from the cells was degraded by electrochemical oxidation. The removal rate of chlorophyll-a could reach 96% at the current density of 10 mA cm(-2). Electrochemical treatment caused minor variation of pH values and conductivity of the suspensions. After electrochemical cycling processes, the optical density at 680 nm of algal cell suspensions remained below 0.1 after 6 days, and it showed that cells had no potential to survive and grow. The results implicated that the inactivation of M. aeruginosa was successfully performed by the electrochemical treatment, and it made the algal cells lose ability to survive, demonstrating the potential of such an alternative process for efficient water purification. PMID- 16047804 TI - Taking the fungal highway: mobilization of pollutant-degrading bacteria by fungi. AB - The capacity of fungi to serve as vectors for the dispersion of pollutant degrading bacteria was analyzed in laboratory model systems mimicking water saturated (agar surfaces) and unsaturated soil environments (glass-bead-filled columns). Two common soil fungi (Fusarium oxysporum and Rhexocercosporidium sp.) forming hydrophilic and hydrophobic mycelia, respectively, and three polycyclic aromatic hydrocarbon degrading bacteria (Achromobacter sp. SK1, Mycobacterium frederiksbergense LB501TG, and Sphingomonas sp. L138) were selected based on the absence of mutual antagonistic effects. It was shown that fungal hyphae act as vectors for bacterial transport with mobilization strongly depending on the specific microorganisms chosen: The motile strain Achromobacter sp. SK1 was most efficiently spread along hyphae of hydrophilic F. oxysporum in both model systems with transport velocities of up to 1 cm d(-1), whereas no dispersion of the two nonmotile strains was observed in the presence of F. oxysporum. By contrast, none of the bacteria was mobilized along the hydrophobic mycelia of Rhexocercosporidium sp. growing on agar surfaces. In column experiments however, strain SK1 was mobilized by Rhexocercosporidium sp. It is hypothesized that bacteria may move by their intrinsic motilitythrough continuous (physiological) liquid films forming around fungal hyphae. The results of this study suggest that the specific stimulation of indigenous fungi may be a strategy to mobilize pollutant-degrading bacteria leading to their homogenization in polluted soil thereby improving bioremediation. PMID- 16047805 TI - Fractionation of UV and VUV pretreated natural organic matter from drinking water. AB - Recent studies have examined the potential of ultraviolet (UV, 254 nm) and vacuum ultraviolet (VUV, 185 nm + 254 nm) irradiation as either a pretreatment for a biological process or as a sole treatment for the removal of natural organic matter as dissolved organic carbon from drinking water. To understand the potential of UV and VUV irradiation followed by subsequent biological treatment, treated water was fractionated into four components: very hydrophobic acid (VHA), slightly hydrophobic acid (SHA), hydrophilic charged (CHA), and hydrophilic neutral (NEU). The VHA fraction was found to be very susceptible to both UV and VUV irradiation, and the fragmentation products of the high molecular weight VHA and SHA molecules contributed to the CHA and NEU fractions to form a pool of biodegradable, non-UV-absorbing, low molecular weight moieties. The NEU fraction was the most difficult to remove, as most of the components in this fraction were refractory to both the biological and photo-oxidative processes. Therefore, enhanced removal of the NEU fraction is required to increase the effectiveness and potential of the treatment process. PMID- 16047806 TI - Contemporary anthropogenic silver cycle: a multilevel analysis. AB - Anthropogenic cycling of silver in 1997 is presented using three discrete governmental units: 64 countries encompassing what we believe to be over 90% of global silver flows, 9 world regions, and the entire planet. Using material flow analysis (MFA) techniques, the country level cycles are aggregated to produce the regional cycles, which are used to form a "best estimate" global cycle. Interesting findings include the following: (1) several silver-mining countries export ore and concentrate but also import silver-containing semiproducts and products; (2) the level of development for a country, as indicated by the gross domestic product, is a fair indicator of silver use, but several significant outliers exist; (3) the countries with the greatest mine production include Mexico, the United States, Peru, and China, whereas the United States, Japan, India, Germany, and Italy lead in the fabrication and manufacture of products; (4) North America and Europe's use of silver products exceed that of other regions on a per capita basis; (5) global silver discards, including tailings and separation waste, totaled approximately 57% of the silver mined; (6) approximately 57% of the silver entering waste management globally is recycled; and (7) the amount of silver entering landfills globally is comparable to the amount found in tailings. The results of this MFA lay the basis for further analysis, which in turn can offer insight into natural resource policy, the characterization of environmental impact, and better resource management. PMID- 16047807 TI - Microbial fuel cell using anaerobic respiration as an anodic reaction and biomineralized manganese as a cathodic reactant. AB - We have operated a microbial fuel cell in which glucose was oxidized by Klebsiella pneumoniae in the anodic compartment, and biomineralized manganese oxides, deposited by Leptothrix discophora, were electrochemically reduced in the cathodic compartment. In the anodic compartment, to facilitate the electron transfer from glucose to the graphite electrode, we added a redox mediator, 2 hydroxy-1,4-naphthoquinone. We did not add any redox mediator to the cathodic compartment because the biomineralized manganese oxides were deposited on the surface of a graphite electrode and were reduced directly by electrons from the electrode. We have demonstrated that biomineralized manganese oxides are superiorto oxygen when used as cathodic reactants in microbial fuel cells. The current density delivered by using biomineralized manganese oxides as the cathodic reactant was almost 2 orders of magnitude higher than that delivered using oxygen. Several fuel cells were operated for 500 h, reaching anodic potentials of -441.5 +/- 31 mVscE and cathodic potentials of +384.5 +/- 64 mVscE. When the electrodes were connected by a 50 Ohms resistor, the fuel cell delivered the peak power density of 126.7 +/- 31.5 mW/m2. PMID- 16047808 TI - [The construction of the pedagogic project: nursing school experience]. AB - Social historical study, which has as object the construction of the pedagogic/political project of the Undergraduate Course of the Nursing School of the Rio de Janeiro State University (FENF/UERJ)--1992-2002. The objective is to report the experience of the construction of the Pedagogic Project. This research was conducted by analyzing documents and dissertations belonging to the 'Dr. Nalva Pereira Calda' Memory Center Collection. It is theoretically based on authors who deal with this theme. The structure and the curriculum plan of the Undergraduate Course were approved by the Superior Council of Teaching and Research of UERJ in December 2001, through the Deliberation no. 05/01, becoming valid as of the first half of 2002. PMID- 16047809 TI - [Appointment with nurses for transplanted heart clients--impact of educative health actions]. AB - The study was aimed at evaluating the impact of educative health actions on nursing appointments with 18 transplanted heart clients, based on Orem's Theory. It was conducted in a Heart Transplant and Insufficiency Unit of a Public Health Institution in Fortaleza-CE. The data were collected by way of nursing appointments. 14 nursing diagnoses were identified in the first appointment and 7 in the last one. It was found out that the clients developed self-care skills with the implementation of educative health actions, tuming them into self-care agents, reaching a good level of health and well-being. On the other hand, it raised nurses' awareness of a systematic and efficacious management of the clinical monitoring of their clientele. PMID- 16047810 TI - [Intensive home-based care: is it a possible reality for the public health system?]. AB - A qualitative investigation characterized as an exploratory study of individual case. The guiding point was inquiring into how the Public Health System (SUS) provides intensive home-based care. The study was aimed at analyzing how this health support is made available--that is, what kind of assistance is given and what is the equipment maintenance and warranty like--and at examining how the subject of this study obtained this service, that is, how the service reached him/her and how long it usually takes to do so. The results show that the paths to get life support are full of obstacles and that this path in search of home based care through the SUS dynamics involves bureaucracy, professionals, and equipment that often make it difficult for the population to have access to public health services. PMID- 16047811 TI - [Health education: object of study in dissertations and theses by Brazillian nurses]. AB - This study was aimed at surveying the quantity of dissertations and theses written by nurses on health education, identifying and analyzing their methodological steps. 17 Catalogs of Information about Research and Researchers from CEPEn/ABEn--published from 1979 to 1999--have been examined. The sample comprises 105 dissertations and 15 theses about the theme. Chronic diseases, pregnancy-puerperal cycle, health education, management and perisurgery situation have stood out as objects of study. The populations were adults, nursing professionals, women, children and adolescents. Interviewing was the most used technique. Authors indicated the need to improve educative practices and considered that health education motivates changes in lifestyle. It was concluded that there is a need to strengthen research into the everyday of Nursing in a historical and social context. PMID- 16047812 TI - [The nurse in the decentralization process of the health system]. AB - Study about the decentralization process of the health system in the '80s and '90s in the city of Itabuna-BA. It is aimed at describing the city's health decentralization process, identifying the nurse's insertion/participation in this process. Descriptive/qualitative study of exploratory nature that utilized both the semistructured interview and documental analysis for data collection. The results show that the nurse's insertion in the decentralization process took place according to the changes that occurred at every municipal management, where the nurse acted more effectively as the set of circumstances was established and was influenced by the several conjunctures formed by the implemented policies or implemented in each context of the management of the municipal health system. PMID- 16047813 TI - [Errors of medication in a university hospital: type, causes, suggestions and actions]. AB - This exploratory study (survey) has analyzed the errors of medication in a university hospital on the basis of 40 interviews conducted with professionals of the medication system. The results showed that the most frequent types of errors occur in drug prescription (29.04%); individual flaws are considered the main cause of errors (47.37%) and the main flaws in the medication system (26.98%); changes in individual attitudes are suggested as a way to avoid errors (28.26%); guidance is the most utilized administrative action (25%). The conclusion is that there is no systemic awareness among professionals regarding errors, putting the blame on the human being. This culture must be altered and transformed into improvements for the system. PMID- 16047814 TI - [Education of nurses in the perspective of the Psychiatric reform]. AB - This study is the result of research conducted by senior students of the Nursing Course on the conceptions of the mental health/disease process in the perspective of the Psychiatric Reform proposed countrywide. It was based on the theoretical methodological system of the Historical and Dialectical Materialism and it has Work as its category of analysis. The empirical material was analyzed through the Speech Analysis technique. The themes gathered from the speeches converged into the formation of an empirical category that refers to the reproduction of the knowledge of traditional psychiatry in the teaching of Nursing, indicating a professional education based on that conservative social position. PMID- 16047815 TI - [The (in)visible gender of old age in the knowledge of nursing]. AB - It is a bibliographical study in which the scientific production of Nursing regarding Women in Old Age was surveyed. The study was based on the Catalog of Dissertations and Theses from the CD-ROM of the Center for Studies and Research on Nursing (CEPEn) of the Brazilian Nursing Association (ABEn), from 1979 to 1999. It was aimed at getting to know the deviations and/or approximations in the knowledge of Nursing by way of the object of study of the Ph.D. thesis 'Health practice and knowledge of old women in the gender perspective'. It was found out that old women have been investigated in few studies. In addition, the objects of these investigations have strongly deviated from gender issues, demonstrating the sexless aspect of old age and the denial of women's social roles. PMID- 16047816 TI - [The influence of the "Friend of Children Hospital" campaign on breast-feeding]. AB - The paper investigates the awareness of women who gave birth in a 'Friend of Children' Hospital (FCH) of the influence of this campaign on their decision to breast-feed. We conducted semi-structured interviews, focusing the maternal breast-feeding process, expectations, experiences, and the importance of the health staff. The data analysis was based on the "Ten Steps to Successful Maternal Breast-Feeding." We found out that in situations regarded by women as problematic and lacking in answers that could attenuate their discomfort and suffering in the face of their desire to breast-feed, they stopped breast feeding. We believe that by maintaining the FCH support to women (so that they can face those difficulties), its influence can be decisive to the continuation of breast-feeding. PMID- 16047817 TI - [Feelings of patients on the waiting list for a kidney transplant]. AB - A descriptive and exploratory study with a qualitative approach whose aim was to get to know the feelings of patients on the waiting list for a kidney transplant. The research was conducted at the Hemodialysis Unit of a university hospital of Porto Alegre. The participants were the active patients on the waiting list for a kidney transplant. The sample comprised 9 patients affected with chronic renal insufficiency who undergo hemodialysis. The following feelings have been identified: hope, anxiety, freedom, ambivalence, fear, guilt, and faith. Dependence on dialysis for survival causes patients to face death and disbelief at the same time that they search for strength and faith in order to fight and wait for a donor. PMID- 16047818 TI - [The prevalence of tuberculosis in the state of Acre]. AB - It is a descriptive study with the aim of studying the prevalence of tuberculosis in the State of Acre from 1995 to 2001. The data were collected from the forms of the Program for Tuberculosis Control of the Health and Sanitation Office. The data show that the incidence has been maintaining an average of 343.7 new cases a year. The pulmonary type was the most prevalent one, accounting for 90.0%. Among the age groups, the 20-49 age range, with 1347 (56.0%) and the 60 and over-60 age range, with 412 (17.0%), stand out. From 1998 to 2001 the male sex was the most attacked one by the disease, when the year 2000 accounted for 58.2% of the cases. The average of the cure rate was 71.0% and abandonment of treatment was 14.8% in 2001 and 32.3% in 1995. The smallest percentage of death occurred in 1995 (1.0%) and the largest one in 1997 (7.0%). Five cases were notified for the occurrence of coinfection between TB/AIDS and HIV/TB. The program's population coverage was within 90% every year. From the 22 cities, 6 (27.3%) still do not have any action for controlling the disease. PMID- 16047819 TI - [Critical/creative nursing professional: the construction of the intersectional space in the pedagogical relation]. AB - This study is aimed at understanding how the intersectional space manifests itself in the pedagogical relation in an Undergraduate Nursing Course. It is based on Merhy's theoretical system, which establishes the intersectional space in the health work process as a moment that can promote the implementation of changes. The utilized methodology is based on a qualitative approach in the style of instrumental case study, using the observation technique in the formal fields of teaching. We have identified a disproportion between the instituted programs and curiosity, reaffirming that the educative process needs to be a knowing act among subjects, in a dialogic relation mediated by words, emotions and knowable objects. We have concluded that the instituting pedagogical relation represents a commitment to the transformation of the health practices. PMID- 16047820 TI - [Reduction in the number of questions of a scale designed for measuring opinions]. AB - In order to study people's opinions on concepts such as mental health, mental illness and assistance in mental health, the Scale for Measuring Opinions (SMO) was built and applied. With the aim of improving the efficiency of this instrument, the present study was conducted on the basis of three criteria: (a) critical analysis of the sentence and of the pertinence of each question, observing repetitions and problems understanding the sentences; (b) importance of the assertion based on the total score, calculating Pearson's correlation coefficients; (c) correlation of each question with the average score per category. Questions have been reformulated, regrouped or eliminated, considering their pertinence or low correlation with the average score. As a result, a 34 item SMO was obtained. PMID- 16047821 TI - [Potentially infectious residues at hemotherapy services and interfaces with infectious diseases]. AB - The aim was to identify the processing of potentially infectious residues and of piercing and cutting residues at hemotherapy units and their possible relation with workers' health. No service presented a structured plan for managing health service residues or for the occupational safety program. We found an occupational accident rate of 32.82% and that 9.16% of the workers did not have a complete plan of vaccination against hepatitis B. In-service training in handling residues and in occupational health are not standardized. The rate of serum conversion for HBV and HCV after occupational exposure was 11.63%. We concluded that the processing of potentially infectious residues and of piercing and cutting residues at the examined hemotherapy units has exposed workers to the acquisition of infectious diseases. PMID- 16047822 TI - [Risk of inffective breast-feeding: a nursing diagnosis]. AB - This study is aimed at presenting the 'risk of ineffective breast-feeding' nursing diagnosis regarding mothers of premature infants who are hospitalized in a neonatal Intensive Care Unit. The case study outlined the methodology of the study, and the sample is composed of 35 mothers. Such a diagnosis was detected in 100% of the sample and the risk factors are prematureness; insufficient opportunity to breast-feed due to the newborn's hospitalization; lack of knowledge regarding the maintenance of lactation; maternal fear; inconstancy of breast suction due to separation; and artificial feeding of the newborn. It is believed that the identification of the risks of ineffective breast-feeding during the newborn's hospitalization period makes possible a nursing care focused on the prevention of an 'ineffective breast-feeding' diagnosis. PMID- 16047823 TI - [Popular wisdom: its existence in the university environment]. AB - Nowadays, myths and superstitions are present in spite of scientific and technological developments, especially when trying to solve problems that escape human understanding. This study was aimed at determining the existence of superstitions and myths in the university community, investigating their origins, influences, adoption and credibility, correlating them with people's level of knowledge. It is a descriptive/analytical research conducted at Teaching Units in the Area of Health of the Federal University of Goias. The technique of content analysis was utilized for data analysis. Two categories have been created: Personal Attitudes related to Superstitions and Influences and Destruction of Superstitions. It was found out that there is a clash between popular and scientific knowledge, either leading to the exclusion of popular wisdom, to its 'veiled' maintenance, or even to an alliance between the two types of knowledge. PMID- 16047824 TI - [The meaning of cancer in the everyday of women undergoing chemotherapeutic treatment]. AB - The aim of this study was to reveal the feelings of women affected with breast neoplasia in their family environment. Its methodological strategy is based on the phenomenological approach, a method that seeks to reveal the phenomenon, that is, that which shows itself to itself, considering the language of the person who experiences it. When questioned about their experience with cancer, women expressed their view, their feelings and experiences with the disease. Two different situations arose from the analyzed speeches: their experience with cancer and the experience of their family members after the diagnosis of cancer. The study led us to realize that each person reacts differently to his/her vicissitudes in life, revealing how painful or pleasurable the events of life are. PMID- 16047825 TI - [Meanings of non-care practices from the point of view of hospital clients]. AB - Its aim is to identify awareness of the types of non-care (lack of care)/violence that occur at hospital units. The methodology is qualitative and we have interviewed 29 clients. We have established four main categories: lack of attention, disrespect, discomfort, and irresponsibility. The results indicate that the lack of care/violence is a Power Problem related to disrespect, prejudice and/or discrimination due to a person's social/sexual condition. It is characterized by rudeness, impoliteness, arrogance, sexual harassment, contempt, and indifference on the part of healthcare professionals. The lack of care/ violence, as an Asymmetric Problem, is characterized by a behavioral attitude of power and by the lack of a horizontal dialogic communication. Lack of attention, interest and will, and discomfort have been reported. We concluded that the lack of care/violence practices occur in the everyday of the interrelations of the care-taking process between client and health professionals. PMID- 16047826 TI - [Nursing appointment (consultation) for the promotion of sexual health]. AB - Methodological bases and instruments are presented in this academic paper for holding a nursing consultation focused on the sexual health of adults and adolescents. The systematized proposal is based on a promotional perspective of sexual health and on a comprehensive concept of sexuality. Emphasis is placed on the recognition of related vulnerabilities, on an ample approach of needs, and on the inter-relational and educative dimension and educative dimension in the work of nurses. PMID- 16047827 TI - [Clarifying the implementation of nursing care systematization]. AB - This study has reviewed the national literature regarding nursing assistance systematization (NAS), with the aim of identifying the difficulties implementing this practice and the factors that interfere with and harm its implementation. The MEDLINE, LILACS, and BDENF databases have been utilized and six studies published in nursing periodicals in the last five years have been surveyed. The results indicate several difficulties implementing the NAS and several factors that interfere negatively with its implementation. Considering the importance of this assistance methodology for valuing professional nursing, it is necessary to reflect on/discuss its practical difficulties so that we can overcome them, making it a pleasurable activity capable of providing nurses with autonomy and providing patients with quality assistance. PMID- 16047828 TI - [Nursing and caring for in the area of mental health]. AB - This study describes the sociopolitical aspects that involved the psychiatric assistance reform, focusing the deinstitutionalization process and the importance of nursing beyond a component of the interdisciplinary team. It aims to analyze the process of nursing assistance to the mentally sick in services outside the hospital. It is a bibliographical review of national periodicals from 1999 to 2001. The authors discuss workers' engagement in the area of mental health, with the "deconstruction"/construction about care, making necessary a humanized approach by way of the interpersonal relationship of patients, nurses and the teams responsible for giving assistance to the mentally sick. PMID- 16047829 TI - [The institutional risk analyst nurse]. AB - The study reports the experience developed by the risk analysis nurse by a process of identification, evaluation and treatment of the professional and institutional risks, to which each hospital is exposed. Besides the liability related to quality, there is another regarding the Professional Liability, related to the work of the professionals within the institution. The data collection is done with the filling up of a form, formal and informal interview with the responsible for the services rendered. The result obtained identifies the potential risk factors, which can generate hospital and professional damages. The nurse elaborates the report, and, as a preventive strategy, lists recommendations. By carefully analyzing each risk, is possible to recommend the best way of managing it. PMID- 16047830 TI - [Violence in the nursing workplace: a new occupational risk]. AB - This study is aimed at presenting a review of the literature regarding violence in the nursing workplace. The main aspects in relation to violence in the nursing workplace that were found in literature are: the types of violence, the prevalence of violence in the workplace; factors associated with the risk; consequences for workers and prevention of these events. Workers are exposed to several types of violence, causing serious consequences to health. It is necessary, therefore, that other studies about this subject matter are conducted, mainly in our country. PMID- 16047831 TI - [AIDS and diabetes mellitus versus distributive justice in the public health system]. AB - Reflection about the way of distributing public resources in the area of health and the ensuing consequences. The fairness of distributive justice is questioned. We propose a contextual reading of the official figures through interrelations that make it possible to understand the meaning of government figures. AIDS and Diabetes, which have been chosen as paradigms, represent, respectively, the groups of Transmitted Diseases and Non-Transmitted Diseases. The official figures relating to the two diseases are correlated in some precise aspects such as expenses on drugs, prevalence, and evolution of mortality rates. We alert to the remarkable differences in official attention between these two diseases and to the consequences suffered by people affected with Diabetes and other nontransmitted diseases, arising from the insufficiency of public resources allotted for their health care. PMID- 16047832 TI - [Endomarketing: Essay on possibilities of innovation in nursing management]. AB - This article is aimed at presenting endomarketing as a tool for nurses in the management process, taking into account the current context of changes in the healthcare sector, which also requires the development of skills that are directly related to the introduction of new technologies and ways of work organization. Our reflection leaves some important points to be analyzed by nurses and educators in the search for better ways and alternatives for educating nurses who will have greater professional satisfaction and competence to manage the healthcare services. The big challenge of endomarketing is, therefore, to combine the aims and interests of the internal public with the needs and expectations of the external public in the organization. PMID- 16047833 TI - [Humanized care in the teaching of nursing]. AB - This 'teaching experience' report addresses contents related to the humanized care given to children/adolescents and to their family. It was made in collaboration with students of the Undergraduate Course in Nursing. The Creativity and Awareness workshops proposed by Cabra I (1999) have been used as methodology because they proved adequate to approaching complex themes such as prevention of violence against children/adolescents and assistance to families that experience child sickness. During the workshops, the aim is to create an environment for reflection on humanized care on the basis of students' experiences. We found out that it is important to introduce alternative methodologies into the teaching/learning process of students, seeking the construction of knowledge and giving priority to their experiences. PMID- 16047834 TI - [Health education: a transforming experience]. AB - This article describes the positive and transforming experience of the 'health education' practice, highlighting the results produced on both educators and the people who participated in the education process. The work took place in 2003 in the city of Manacapuru-Amazonas, and it consisted of educational activities designed for teenagers from a public school of the city The educative activities were based on a methodology for nursing assistance to collective health, using its phases. The results showed, in fact, that health education is essential to the reflection and change of behavior in people's lives. Therefore, health education must be systematically planned and adopted as an important role of the nursing professional. PMID- 16047835 TI - [The historical prccess of the nursing work in the city of Cuiaba-Mato Grosso]. AB - The study is a historical research whose aim was to historicize the singularity of the nursing work process in the city of Cuiaba/MT The data were collected from primary and secondary sources. According to the analyzed data, nursing appeared with the attention and care for the sick, and volunteers, blacks and slaves were the ones who performed it. The first created institution was the Santa Casa de Misericordia of Cuiaba. The religious, lay people and slaves performed nursing assistance on the basis of abnegation, friendliness, and godfathering. Successive important events such as the implementation of ABEn/MT, the Undergraduate Course in Nursing, and COREN/MT have marked the course of the nursing work, the three of which were linked to the business community in the health sector in the studied context. PMID- 16047836 TI - [34,450 days of cardiac surgery in the Czech Republic]. AB - Professor Karel Maydl was born in Rokytnice nad Jizerou, where his father was employed at the local court. Czech cardio-surgery was born with the first successful treatment of the stab in the heart, which was done by professor Rychlik in 1910. The next few years the cardio-surgery was represented only by occasional treatment of heart injury. Only in 1934 Jiri Divis excided ganglion stelatum to cure angina pectoris and in 1936 he performed surgical treatment of constrictive pericarditis. Article reviews all Essentials moments in the development of cardio-surgery in Bohemia and Moravia. The second part of the paper gives at present the most frequently performed surgeries. It is concluded that the Czech cardio-surgery has according to the numbers and duality of the treatment the west-European level. At the end Professor Maydl, who died for heart failure, is quoted, why he himself had to be stroked by the disease, which is not possible to treat with the knife. In the 100 years since that it is possible to evaluate that in ICEM 283 medical doctors have been treated for the heart disease and results of their operations were Berger than in standard population. It reproves the bad record of doctors to be bad patients. PMID- 16047837 TI - [Proteomics and its role in medicine]. AB - Proteomics is a new methodological and conceptual approach to the study of live systems, which aims to map the whole proteom - the protein complement of genome. It aspires to describe quantitatively and qualitatively all proteins present at the given moment in the cell, tissue or the organism. The principal tools of proteomics are separation techniques based on electrophoresis and chromatography, which are used to fractionate complex protein mixtures and namely the mass spectrometry used for the identification of individual proteins. Proteomics represents very young and rapid developing specialization, which has already proved its valuable potential for the study of various physiological and pathlogical molecular mechanisms and for the direct use in the diagnostics of serious diseases. PMID- 16047838 TI - [Exercise tolerance in patients with chronic obstructive pulmonary disease]. AB - Limitation of exercise tolerance, especially activities of daily living, is the most significant clinical experience, which greatly affects quality of life of patients with chronic obstructive pulmonary disease (COPD). Many advances in the understanding of the pathophysiological mechanisms of bronchial obstruction in patients with COPD and their meanings for diagnosis and monitoring of the disease have occurred during the last two decades. The author discusses the most significant factors, which influence tolerance of physical exercise in patients with more advanced forms of COPD, and brings the attention to a practical test of physical capacity. PMID- 16047839 TI - [The role of pentraxin 3 in the inflammatory and immune response]. AB - Pentraxin 3 is the first detected and so far the most important protein from the recently recognized group called the long pentraxins. The structure and function of PTX3 resembles in many aspects that of the short or classical pentraxins, i.e. C-reactive protein and the serum amyloid P component. There are, however, several important differences between the two groups of pentraxins that will be mentioned in more detail in the article. All of the above mentioned pentraxins take an acitve part in the first-line defense of the host against invading pathogenic microorganisms and in the clearance of the host's own apoptotic cells. The latter mechanism impedes the onset of destructive autoimmune reactions. A biologically relevant antipode of PTX3 is represented by TNFalpha. Physiologic course of the defense reactions depends on a closely co-ordinated activity of both peptides. In case of an unchecked or missing activity of either peptide, a disturbance in their mutual balance results in increased susceptibility of the host to conditionally pathogenic fungi or in increased damage to host's own tissues inflicted by the defense reactions. This review article deals with the physiopathologic importace of pentraxin 3 as has been gained on the basis of the most up-to-date information. PMID- 16047840 TI - [Advanced glycation and oxidation products in patients with atherosclerosis]. AB - BACKGROUND: Oxidative stress can potentiate atherogenesis via modification of biological structures and formation of new compounds, e.g. advanced glycation end products (AGEs) and advanced oxidation protein products (AOPP). The aim of the study was to determine AGEs and AOPP in patients with atherosclerosis, effect of statin therapy and relationship to parameters of lipid metabolism. METHODS AND RESULTS: AGEs (carboxymethyllysine - ELISA and fluorescent AGEs - spectrofluorimetry) and AOPP (spectrophotometry) were assessed in 42 patients with atherosclerosis and 21 healthy controls. AGEs are significantly elevated in patients with atherosclerosis in comparison with healthy subjects (carboxymethyllysine 9.02+/-1.66 microg/g prot. vs 7.52+/-1.18 microg/g prot., p<0.001, fluorescent AGEs 4.39 x 103+/-1.15 x 103 AU/g prot. vs 3.78 x 103+/-0.52 x 103 AU/g prot., p<0.001). Mean AOPP concentrations are also slightly higher, but this elevation is not quite significant (95.0+/-42.9 micromol/l vs 79.7+/ 28.2 micromol/l, p=0.096). AGEs and AOPP correlate significantly with each other and with selected lipids. Patients with atherosclerosis treated with statins have slightly lower CML, AGEs and AOPP (it did not reach the statistical significance). CONCLUSIONS: Advanced glycoxidation products are elevated in patients with atherosclerosis and are related to parameters of lipid metabolism. Glycoxidation might be possibly therapeutically influenced by statins; however, further clinical studies are required to confirm this hypothesis. PMID- 16047841 TI - [Mutations in the HFE gene in patients with rheumatic diseases]. AB - BACKGROUND: Hereditary hemochromatosis is one of the most common autosomal recessive diseases. Aim of the study. 1. To establish frequency of C282Y and H63D mutations in the HFE gene (the hemochromatosis gene) in general population of the Czech Republic and in patients with hemochromatosis. 2. To find out whether hemochromatosis in homo- or heterozygous state plays a role in the pathogenesis of rheumatic diseases. METHODS AND RESULTS: In 32 patients with hereditary hemochromatosis, in 84 patients with polymyositis or dermatomyositis, in 246 patients with juvenile idiopathic arthritis and in 481 persons of the control group the presence of HFE gene mutations was etablished. The HFE gene mutations were screened for by restriction enzyme analysis performed on PCR amplified products. In the control group, 6.86% carriers of the C282Y mutation and 26.61% those of H63D were found. Homozygous C282Y or H63D mutation was found in 90.6% (p<0.001) of patients with hemochromatosis. Heterozygous C282Y mutation was found in 12.2% (p<0.05) of patients with juvenile idiopathic arthritis. We didn't detected higher prevalence of HFE gene mutations in patients with polymyositis and dermatomyositis. CONCLUSIONS: Results of this study show that heterozygosity for C282Y mutation may be a risk factor for juvenile idiopathic arthritis but not for polymyositis and dermatomyositis. PMID- 16047842 TI - [Prevalence of the overweight and obesity in the random selected group of young females and their clinical-somatic parameters]. AB - BACKGROUND: Obesity is serious health-nutritional problem and its incidence moves into younger age. The aim of the study was to evaluate clinical - somatic parameters of the health-nutritional state in the random selected group of young females. Prevalence of the overweight and obesity were studied and risk of their reciprocal relations of the clinical - somatic parameters was evaluated. METHODS AND RESULTS: We evaluated 161 females between the ages of 20-25 years (average age 21,6+/-1,1 years). The body fat content was measured using the method of bioelectrical impedance analyse and Bodystat Quadscan 4000 (Bodystat Ltd, Doubles, Isle of Man, UK). Body mass index, index of centrality, waist to height ratio, ideal body weight and difference between the actual and ideal body weight was calculated on the basis of measured values. Total cholesterol in the blood was measured by Reflotron Plus (Matsuka Co. LTD, Japan). The results showed that in the tested group of females 49,1% body fat content had less than 20% (group A), 47.8% women had body fat content varying from 20 to 29% (group B), and 3,1% had fat content varying from 30-34% (group C). The higher values almost of all evaluated parameters was confirmed (total cholesterol, body mass index, waist circumference, index of centrality, waist to height ratio and difference between the actual and ideal body weight) in groups with higher fat content (group B and C). In all groups significant differences were observed. According to circumference of waist and hip the android health risk had 3,1% of women. The increased risk of metabolically complications of obesity had 5,6% of women and high risk 1,2% of women. The critical value of cholesterol had 8,1% of investigated women. Accorging to the body mass index (D, E, F groups) had 90,1% of females normal body weight (group D), 9.3% of overweight (group E) and only 0.6% obesity. With increasing value of body mass index were also increased values of all monitored parameters, between groups D and E was differences significant in six of eleven parameters (body weight, waist circumference, waist to hip ratio, waist to height ratio, difference between the actual and ideal body weight). CONCLUSIONS: The increased cholesterol level is more frequent in women body with higher content of body fat. Groups with higher content of body fat have, except of body weight and body mass index, concurrently higher the waist circumference, waist to hip ratio, waist to height ratio and difference between the actual and ideal body weight. PMID- 16047843 TI - [Takotsubo cardiomyopathy: case report and literature review]. AB - The authors present an interesting case report of 69-year-old caucasian woman with Takotsubo cardiomyopathy. Takotsubo cardiomyopathy is a relatively recently described heart syndrome that probably develops due to the direct toxic effect of excessively released catecholamines on cardiac adrenoceptors during emotional or physical stress. The typical features include reversible left ventricular apical dyskinesis, chest pain with ST-T changes on ECG, minimal myocardial enzymatic release and the absence of coronary stenosis on coronary angiogram. Early coronary angiographic examination is highly recommended as the clinical picture of this syndrome mimics acute myocardial infarction. Betablockers are considered to be the treatment of choice. PMID- 16047844 TI - [Traffic accidents from the view of neurologist]. AB - Review article deals with traffic accidents in our country and the problems of their causes. In comparison with economically developed countries traffic accidents prevalence raised since 1990 till 1995 and with small decline in 2004 it remains on a high level of 140 deaths in traffic accidents for I million of inhabitants, which makes the Czech Republic one of the worst among the compared countries. In agreement with present views, the main course of accidents is the human error. Attention is given to traffic accidents prevalence of young men versus young women, and to the evaluation of psycho-physiologic functions in elderly drivers. Personal qualities are among the most important in the neglecting of traffic rules, in the aggressivity, non-tolerance and disrespect to the freedom of others. Besides the building of the more perfect infrastructure, it is necessary to improve the register of accident recidivist drivers and for accidents and repeated accidents to promote compulsory psycho-neurological examination. Similarly to preventive check outs for alcohol, it s necessary to introduce similar measures to cope with growing effects of drugs. PMID- 16047845 TI - [What to expect from the pathologist in the diagnostics of colorectal cancer? Experience from the clerkships in the US]. AB - Nowadays, surgery still remains the basic treatment modality in the therapy of colorectal cancer. An exact pathological evaluation of the extent of tumour spread and the quality of the resection is also essential for indication of adjuvant therapy and for the assessing prognosis. In the treatment of colorectal cancer, it is precisely this collaboration between the surgeon and pathologist which is fundamental to the multidisciplinary approach. The pathologic examination must concentrate on the macro- and microscopic pathological anatomy, evaluation of lymph node positivity, and presence of micrometastases or other signs of local tumour progression. All these factors can help to establish the prognosis. The standards of pathological examination and methods differ between centres and countries. From personal experience during clerkships in the United States the authors offer an overview of how American pathologists approach examination of colorectal cancer, and which characteristics of the disease are concentrated upon. PMID- 16047846 TI - [Alcoholic liver disease]. AB - This paper summarizes the mechanisms which cause alcoholic liver disease. The most important principle when treating alcoholic liver disease is to abstain from alcohol; this often brings about remarkable improvement. Alcoholic liver disease should be treated as early as possible. PMID- 16047847 TI - [Evaluation of gastroenterology procedures by general practitioners in the Czech Republic--contribution to interdisciplinary cooperation]. AB - BACKGROUND: The understanding of different management strategies in primary and secondary care is important for cooperation between specialists and general practitioners in clinical practice. The Institut of General Practice at First Faculty of Medicine carried out a survey for gastroenterologists concerning their opinions on general practitioners and their current knowledge, competence and management of GI disorders as a contribution to interdisciplinary communication. METHODS AND RESULTS: Questionnaires were sent to 552 gastroenterologists in the country. There was a response rate of 31% without any reminder, 170 questionnaires were included in a study. The most of gastroenterologists (92%) support practical guidelines development. There is an insufficient confidence that these guidelines are implemented in practice. Four from five gastroenterologists consider knowledge of their colleagues GPs in gastroenterology as average. In particular cases (irritable bowel syndrome, step up and step down approach) are specialists rather sceptic. There is a low support for the increase of GP competence in gastroenterology, as for urea breath test availibility, particularly in private gastroenterologists (p=0,04). CONCLUSIONS: Results of the survey could facilitate the interdisciplinary communication in gastroenterology, contribute to CME programs development and may serve as an example for other specialities. PMID- 16047848 TI - A report from the president ... Janet Harris MNA president outlines busy agenda for the association. PMID- 16047849 TI - Noted speaker headlines MNA convention. PMID- 16047851 TI - CMS's hospital compare makes hospital quality available to the public. PMID- 16047850 TI - Board of nursing offers new service. PMID- 16047852 TI - State legislation and advanced practice registered nurses. PMID- 16047853 TI - SHIPS awarded 31.7 million dollars to help beneficiaries about new drug coverage. PMID- 16047854 TI - [Preoperative evaluation of the feasibility of laparoscopic cholecystectomy]. AB - AIM: To determine the possibility of preoperative evaluation of the feasibility of laparoscopic cholecystectomy based on the standard preoperative examinations and findings. METHODS: During 1997, 100 consecutively operated patients with the diagnosis of chronic calculous cholecystitis were followed up. Sex and age, and the results of blood count, sedimentation rate, ultrasonography (US), and intravenous cholangiography (IVC) were monitored. Based on adhesions, fibrosis in Calot's triangle and pericholecystitis, surgical interventions were classified as minor and major. RESULTS: Minor operations were performed in 57, and major in 43 patients. Earlier surgical interventions had been carried out in 8 (18.6%) patients from the major surgery group, while 9 (15.79%) patients had undergone minor surgery. Out of 57 patients with minor surgery, IVC verified the contrast medium filling of the gallbladder in 55 (96.49%) of the patients. Tense gallbladder or wall stratification was not revealed by ultrasonography in any of the patients from this group. Out of 43 patients with major surgery, the gallbladder was not filled with the contrast medium during IVC in 34 (79.07%) patients, while the stratified and tense gallbladder was found by US in 2 (4.65%) patients. The mean sedimentation rate was 14.3 in the patients with minor surgery, and 23.5 in major surgery group. Mean WBC in the patients with minor surgery was 7.4 x 10(9). The patients with major surgery had slightly increased mean value of the white cell count. It was 8.3 x 10(9). CONCLUSION: Statistically significant difference (p < or = 0.05) was found between the variables of the IVC, sedimentation rate, the white blood count, and the earlier operations. No significant difference was found between other analyzed variables. PMID- 16047855 TI - [Microvascular osteoseptocutaneous radial flap in reconstruction of mandible following war injury]. AB - BACKGROUND: Vascularized osteoseptocutaneous radial flap is commonly used in the reconstruction of composite bony and soft tissue defects of the lower third of the face due to the outstanding quality of its cutaneous component. The aim was to evaluate the primary and overall success in the reconstruction of mandibular defects, following war injuries, with vascularized osteoseptocutaneous radial flap. METHODS: At the Department of Maxillofacial Surgery of the Military Medical Academy Belgrade, there were eight patients with this kind of defect following war injury, and the mandible was reconstructed with a vascularized osteoseptocutaneous radial flap. Bony compartment of the graft was harvested as up to 11 cm long segment of radial circumference. RESULTS: The localization and structure of the defect, features of a harvested compound graft, the procedure of the reestablishment of the mandibular continuity was presented as well as immediate and late complications during the consolidation period, and the primary successful reconstruction in 87.5% of the patients. CONCLUSION: The primary and overall success in the mandibular defects reconstruction with a vascularized osteoseptocutaneous radial flap was equal or even better than those presented in the literature on the reconstruction of the similar defects after tumor resections. PMID- 16047856 TI - Comparison of two therapeutic protocols in patients with antiphospholipid antibodies and recurrent miscarriages. AB - AIM: To compare the effects of two therapeutic protocols for the patients with recurrent miscarriages associated with the presence of antiphospholipid (anticardiolipin) antibodies. METHODS: A prospective observational study included 20 patients with antiphospholipid antibodies in the first group who received low molecular heparin and aspirin. The second group of 20 patients, in addition to this therapy, received immunotherapy (intravenous immunoglobulin). Aspirin was administered at the time of a positive pregnancy test, and low-molecular heparin not before the fetal heart activity registration by ultrasound. Intravenous immunoglobulin was given prior to the conception or at the beginning of the pregnancy. We compared these groups according to the pregnancy outcomes and the occurrence of complications during pregnancy, using standard statistical tests. RESULTS: The rate of positive gestational outcome in the patients treated with aspirin and low-molecular heparin was 85% (17/20), and in the second group it was 90% (18/20). There was no significant difference in pregnancy outcomes between these groups (p > 0.05), except for the occurence of preeclampsia and thrombocytopenia, which were recorded only in the aspirin and low-molecular heparin group, but with no statistical significance (p > 0.05) compared to the second group, which received immunoglobulin additionally. CONCLUSION: There was no significant difference (p > 0.05) in pregnancy outcomes between the two studied therapeutic protocols, but the therapy with aspirin and low-molecular heparin was cheaper and easier to apply than the therapy with immunoglobulins. The results of our study confirmed that the final pathogenic mechanisms in recurrent fetal miscarriages were inflammation and thrombosis of the uteroplacental blood vessels. PMID- 16047857 TI - [Reliability of the individual age assessment at the time of death based on sternal rib end morphology in Balkan population]. AB - This paper analyzes the reliability of the Iscan's sternal rib-ends phase method for the assessment of individual age at the time of death in the Balkan population. The method is based on the morphological age changes of the sternal rib ends. The tested samples consisted of 65 ribs from autopsy cases in the Institute for Forensic Medicine, University of Belgrade, during 1999-2002 (23 females, and 42 males of various ages, ranged from 17-91 years), according to the forensic documents. Significant differences between the real chronological age of the individuals and the values established by the Iscan's method was found, especially in the older categories (phases 6 and 7), in both males and females. The results of the discriminative analysis showed the values of the highest diagnostic relevance for the assessment of age in our population: the change of the depth of the articular fossa, the thickness of its walls, and the quality of the bones. PMID- 16047858 TI - [Significance of crown-down root canal preparation technique in endodontic therapy by using the hydroxylapatite sealer]. AB - AIM: To evaluate the crown-down preparation technique, and the use of hydroxylapatite based material for the definitive root canal obturation. METHODS: The investigation included 20 single-canal roots with chronic periapical inflammatory lesion. Biomechanical medicamentous canal preparation was done using the double-flared technique with balanced force, and the obturation was performed with hydroxylapatite sealer (unicone technique). Clinical and radiographic check up performed 12 months after the treatment, used the following parameters: pain, swelling, percussion and palpation sensitivity, and the presence of fistula. RESULTS: The obtained results showed a successful treatment in 18 cases, while in the 2 cases the treatment failed. CONCLUSION: These findings suggested that the crown-down preparation technique efficiently cleaned and shaped the root canal, and that the hydroxylapatite-based material created the homogenous and hermetic root canal obturation, so this methodology could be recommended for the endodontic therapy. PMID- 16047859 TI - [Assessment of feasibility of formulation of a preparation for growth inhibition of metronidazole-resistant strains isolated from periodontal pockets]. AB - INTRODUCTION: Topical metronidazole benzoate preparations, applied directly into the space between the teeth and, periodontal tissue, are the treatment of choice for periodontal pockets. Oleogels and hydrogels, as the biomucoadhesive bases for the manufacturing of these preparations, were developed in our previous trials. The aim of our latest research was to develop a preparation that would solve the problem of inhibiting the metronidazole-resistant anaerobic strains, isolated from the periodontal pockets. METHODS: Metronidazole-resistant strains were isolated from the periodontal pocket swabs, using the established susceptibility testing. Further testing of the antimicrobial activity of the prepared formulations was performed by the membrane-free diffusion method in agar gel. Oleogels and hydrogels were tested with: tetracycline (2.5%), metronidazole (25%), and metronidazole benzoate (40%), as well as with oleogels and hydrogels containing the combination of 2.5% tetracycline and 40% metronidazole benzoate. RESULTS: Satisfying results were achieved with the preparation containing 2.5% tetracycline along with metronidazole benzoate. The main fact revealed by this study was that there was no synergism in tetracycline and metronidazole benzoate activities, if the strain was susceptible to both active substances. CONCLUSION: The best results in the inhibition of the growth of metronidazole-resistant anaerobic strains were obtained by 2.5% tetracycline and metronidazole benzoate preparations in oleogel base. PMID- 16047860 TI - [Urban noise and arterial hypertension]. PMID- 16047861 TI - [Biochemical, pharmacological and toxic effects of n-metil 3,4 methylenedioxyamphetamine--"ecstasy"]. PMID- 16047862 TI - [Indications for the making of porcelain veneers]. PMID- 16047863 TI - [Gangrene of the right colon after blast injury caused by abdominal gunshot wounds]. AB - AIM: To present a patient with an indirect secondary non-perforating blast injury of the right colon following abdominal gunshot injury, which led to necrosis and the right colon gangrene, and was surgically managed. CASE REPORT: A 26-year-old male was shot in the abdomen by four projectiles causing the secondary indirect blast injury of the right colon that turned into gangrene after 24 hours. Two days after admission, laparotomy was performed, but the primary anastomosis was not done because of the stomach and pancreatic injury, and the resection of the colon with terminal ileostomy was done instead. Three months later, the reconstruction of the colon was performed using ileocolotransverso terminolatetral anastomosis. CONCLUSION: Secondary blast injuries should be anticipated in gunshot injuries, and could be expected to any organs, particularly the air filled ones. PMID- 16047864 TI - [Reversible brain damage following acute poisoning with an organic solvent determined by magnetic resonance]. AB - INTRODUCTION: Acute exposure to the effects of volatile solvents is characterized by the abrupt onset of symptoms and signs of poisoning, and relatively fast recovery in the majority of cases. CASE REPORT: We report a 24-year-old patient with an acute, accidental poisoning with a mixture of volatile organic solvents (most probably toluene, styrene and xylene), which led to the development of upward gaze paresis, diplopia, hemiparesis, ataxic gate, and the late onset truncal ataxia episodes. After 6 weeks, he recovered completely, while his extensive brain MRI lesions in the caudate nuclei, laterobasal putaminal regions, bilateral anterior insular cortex, central midbrain tegmental area) withdrew completely after 4 months. CONCLUSION: Acute toxic encephalopathy should be a part of the differential diagnosis in any patient with acute neurobehavioral and neurological deficit. PMID- 16047865 TI - [Aneurysmal bone cyst of the frontal bone]. AB - BACKGROUND: Aneurysmal bone cyst (ABC) is a benign, expansive, osteolytic lesion that mainly occurs in young people, and involves the skull bones only exceptionally. The origin of ABC is controversial: secondary reactive bone lesion, or primary disease that represents an independent nosological entity. Blunt head trauma was suggested as a possible etiological factor. CASE REPORT: A case of a 19-year-old man with primary ABC of the right frontal bone was reported. The lesion was totally excised through frontal craniotomy, and the skull bone defect primarily reconstructed with an acrilate cranioplasty. Five years after the surgery, the patient was without signs of local recurrence. CONCLUSION: Clinical and neuroradiological presentation of the skull ABC was not specific. Pathohistology confirmed the diagnosis. Total excision was the treatment of choice. PMID- 16047866 TI - [From the history of the Military Medical Academy: VIII--radiology, physical therapy, and pharmacy]. PMID- 16047867 TI - The taxman cometh. PMID- 16047868 TI - An open label study of clobetasol propionate 0.05% and betamethasone valerate 0.12% foams in the treatment of mild to moderate acne keloidalis. AB - Acne keloidalis (AK) is a disease affecting primarily African American men. Topical steroids are a widely accepted treatment of AK; however, no studies have been published investigating their effectiveness. The purpose of this open-label study was to assess the efficacy and tolerability of clobetasol propionate 0.05% and betamethasone valerate 0.12% foams in the treatment of AK in 20 African American patients. These patients were treated for 8 to 12 weeks using a pulsed dose regimen. We found topical clobetasol propionate foam to be effective in improving AK, and our patients found the foam vehicle to be cosmetically acceptable. PMID- 16047869 TI - Sarcoidosis mimicking lipodermatosclerosis. AB - The clinical presentation of cutaneous sarcoidosis is highly variable. Rare presentations include ulcerated plaques, morpheaform lesions, and unilateral lower extremity edema. We report the case of a woman who presented with unilateral ulcerating sarcoidosis of the lower leg with progressive fibrosis and edema mimicking lipodermatosclerosis. This case is unique in that the patient exhibited all 3 of the rare manifestations of sarcoidosis; to our knowledge, this presentation has not been previously reported in the literature. PMID- 16047870 TI - A case of calciphylaxis and chronic myelomonocytic leukemia. AB - A 70-year-old woman presented for evaluation of symmetric necrotic ulcers of the lower extremities. Biopsy results revealed changes consistent with calciphylaxis. The predisposing factors in this patient included calcium supplementation, obesity, female gender, viscous blood, renal failure, and diabetes mellitus. To our knowledge, this is the first report of calciphylaxis occurring in the setting of chronic myelomonocytic leukemia. We discuss the history, clinical presentation, diagnosis, and treatment of calciphylaxis. PMID- 16047871 TI - Warfarin-induced leukocytoclastic vasculitis. AB - Skin reactions associated with oral coumarin-derived anticoagulants are an uncommon occurrence. Leukocytoclastic vasculitis (LV) is primarily a cutaneous small vessel vasculitis, though systemic involvement may be encountered. We report 4 patients with late-onset LV probably due to warfarin. All 4 patients presented with skin eruptions that developed after receiving warfarin for several years. The results of skin lesion biopsies were available in 3 patients, confirming LV Cutaneous lesions resolved in all patients after warfarin was discontinued. In 2 of the 4 patients, rechallenge with warfarin led to recurrence of the lesions. LV may be a late-onset adverse reaction associated with warfarin therapy. PMID- 16047872 TI - Papulonecrotic tuberculid: a rare form of cutaneous tuberculosis. AB - We describe a case of papulonecrotic tuberculid, a rare form of cutaneous tuberculosis, in a 25-year-old Philippino woman who had immigrated to Canada 8 years previously. The patient presented with a 3-week history of tender left cervical adenopathy; 1 week later, she developed multiple ulcerated erythematous nodules and emboluslike lesions scattered over her fingers. Results of a biopsy performed on the lymph node revealed granulomatous lymphadenitis, and Mycobacterium tuberculosis grew from the lymph node. Histopathologic analysis of an ulcerative finger lesion demonstrated nonnecrotizing granulomas with dense lymphocytic inflammation of the superficial dermis; however, results of acid-fast staining, mycobacterial culture, and polymerase chain reaction for M tuberculosis complex were all negative. Different conditions can mimic papulonecrotic tuberculid. Therefore, the diagnosis can be difficult unless M tuberculosis is isolated from a site other than the skin, because stain and culture results from skin biopsy specimens are typically negative and the polymerase chain reaction is positive in only 50% of cases. We review the epidemiology, clinicopathologic features, and differential diagnosis of papulonecrotic tuberculid. Awareness of this entity is important to distinguish it from other conditions and to institute appropriate therapy in a timely fashion. PMID- 16047873 TI - Frictional asymptomatic darkening of the extensor surfaces. AB - Frictional asymptomatic darkening of the extensor surfaces (FADES), also known as hyperkeratosis of the elbows and knees, is commonly seen by dermatologists but has never been well characterized. Patients present with uniform, asymptomatic, brown darkening over the extensor surfaces of the elbows and knees with minimal scaling. Both frictional stress and family history may play a role in the pathogenesis of this condition. The results of cutaneous biopsy specimens typically reveal hyperkeratosis, acanthosis, and mild papillomatosis with minimal inflammation. Keratolytic agents such as lactic acid and urea cream along with avoiding frictional stress can be effective in the management of this condition. We describe a series of cases of FADES and its etiology and management options. PMID- 16047874 TI - Combination sodium sulfacetamide 10% and sulfur 5% cream with sunscreens versus metronidazole 0.75% cream for rosacea. AB - Topical metronidazole and combination sodium sulfacetamide and sulfur commonly are used to treat rosacea. Recently, the relative efficacy and safety of sodium sulfacetamide 10% and sulfur 5% cream with sunscreens (Rosac Cream) (n = 75) and metronidazole 0.75% cream (Metrocream) (n = 77) were compared in an investigator blinded, randomized, parallel-group study at 6 sites. After 12 weeks of treatment with sodium sulfacetamide 10% and sulfur 5% cream with sunscreens, there was a significantly greater percentage reduction (80%) in inflammatory lesions compared with metronidazole 0.75% cream (72%)(P = .04), as well as a significantly greater percentage of subjects with improved erythema (69% vs 45%, respectively; P = .0007). In addition, the sodium sulfacetamide 10% and sulfur 5% cream with sunscreens group had a significantly greater proportion of subjects with success in global improvement at week 12 compared with the metronidazole 0.75% cream group (79% vs 59%, respectively; P = .01). There was no significant difference between treatment groups in the percentage of subjects with improvement in investigator global severity. Overall tolerance was good or excellent in 85% of subjects in the sodium sulfacetamide 10% and sulfur 5% cream with sunscreens group and in 97% of subjects in the metronidazole 0.75% cream group. Seven subjects had poor tolerance to the sodium sulfacetamide 10% and sulfur 5% cream with sunscreens, possibly caused by a sulfa drug allergy. PMID- 16047875 TI - Application of TLC for confirmation and screening of pesticide residues in fruits, vegetables, and cereal grains: Part 2. Repeatability and reproducibility of Rf and MDQ values. AB - This paper illustrates the effect of major factors influencing the reproducibility of thin layer chromatography (TLC) separation and detection under largely differing environmental and laboratory conditions. The optimum conditions for reproducibility and detection sensitivity was obtained on 20 x 20 cm layer in the retention factor (Rf) range of 0.2-0.7 by applying the sample in spots of 3-4 mm diameter at 2 cm from the edge of the plate. The reproducibility rapidly decreased below Rf = 0.2. Above Rf = 0.2 the within-laboratory reproducibility of 219 pesticides obtained in ethyl acetate silica gel elution system by several laboratories was typically below 10%. The among-laboratories reproducibility of the average retention factors was generally below 12%. The minimum detectable quantities (MDQ) of 219 pesticide residues were determined with nine detection methods. The MDQ values largely varied depending on the mode of detection. Bioassay methods enabled the detection down to 0.1-10 ng, while 20-100 ng could be achieved with the chemical reagents. Higher MDQ values are also reported in order to assist the identification of compounds potentially present. The between laboratories reproducibility of MDQ values was typically 1-5 MDQmin. PMID- 16047876 TI - Optimization of a methodology for the determination of organochlorine pesticides in surface water by SPME-GC/MS. AB - In the present work experimental conditions were optimized for the analysis of organochlorine traces in water matrix using solid-phase microextraction (SPME) followed by gas chromatography with mass selective detector. The parameters including time of exposure of the fiber in the aqueous sample, fiber type, agitation speed, pH, ionic forces, temperature of adsorption, and time of desorption were evaluated. The best conditions to analyze organochlorine were obtained by using higher than room temperature, agitation of the sample, extraction time of 40 min, and polyacrylate fiber. PMID- 16047877 TI - Phototransformation of clodinafop-propargyl. AB - Photodegradation of the herbicide clodinafop-propargyl was investigated on glass surface under sunlight and UV light. Four photoproducts were identified by NMR, IR, and MS. Major photolysis products were 2-[(5-chloro-3-fluoro-2-pyridyloxy) phenoxy] propanoic acid and prop-2-ynyl-2-[(5-chloro-3-hydroxy-2-pyridyloxy) phenoxy] propanoate, while minor were ethyl 2-[(5-chloro-3-fluoro-2-pyridyloxy) phenoxy] propanoate and 1-hydroxypropanyl-2-[(5-chloro-3-fluoro-2-pyridyloxy) phenoxy] propanoate. Rate of photodegradation followed first-order kinetics with significant correlation coefficient. The major photoproducts were observed in maximum quantity on the 7th and 10th day and further degraded within 15-20 days. PMID- 16047878 TI - Artificial sweat enhances dermal transfer of chlorpyrifos from treated nylon carpet fibers. AB - The dermal transfer and absorption of 14C-ring-chlorpyrifos from nylon carpet fibers was measured in skin penetration-evaporation cells with excised pig skin. Prior to application, synthetic sweat was applied to skin in half of the cells. Radioactivity was measured in receptor fluid, dermis, epidermis, tape stripping samples, and vapor trap samples during a 24-h period. The sum of radiolabel recovered from the dermis and receptor fluid represented the absorbed dose. There was no significant difference (p > 0.05) in percutaneous absorption between cells that received the synthetic sweat application and "dry" cells (1.3 +/- 0.3% of applied dose). There was significantly more (p < 0.05) radiolabel recovered from tape stripping (5.4 +/- 2.1 vs. 2.8 +/- 0.6%) and in the epidermis (4.5 +/- 0.8 vs. 3.1 +/- 0.3%) from cells that received the synthetic sweat application, which indicated synthetic sweat facilitated transfer of chlorpyrifos from a treated substrate to the skin surface. The measured value for percutaneous absorption of chlorpyrifos agreed with the value predicted from an empirical model previously developed for nitro compound-containing soil. PMID- 16047879 TI - Adsorption characteristics of 2,4-dichlorophenoxyacetic acid (2,4-D) from aqueous solution on powdered activated carbon. AB - The removal of 2,4-dichlorophenoxyacetic acid (2,4-D), one of the most commonly used phenoxy acid herbicides, from aqueous solution was studied by using acid washed powdered activated carbon (PAC) as an adsorbent in a batch system. Adsorption equilibrium, kinetics, and thermodynamics were investigated as a function of initial pH, temperature, and initial 2,4-D concentration. Powdered activated carbon exhibited the highest 2,4-D uptake capacity of 333.3 mg g(-1) at 25 degrees C and an initial pH value of 2.0. Freundlich, Langmuir, and Redlich Peterson isotherm models were used to express the equilibrium data of 2,4-D depending on temperature. Equilibrium data fitted very well to the Freundlich equilibrium model in the studied concentration range of 2,4-D at all the temperatures studied. Three simplified models including pseudo-first-order, pseudo-second-order, and saturation-type kinetic models were used to test the adsorption kinetics. It was shown that the adsorption of 2,4-D on PAC at 25, 35, and 45 degrees C could be best fitted by the saturation-type kinetic model with film and intraparticle diffusions being the essential rate-controlling steps. The activation energy of adsorption (EA) was determined as--1.69 kJ mole(-1). Using the thermodynamic equilibrium coefficients obtained at different temperatures, the thermodynamic constants of adsorption (deltaG degrees, deltaH degrees, and deltaS degrees) were also evaluated. PMID- 16047880 TI - Chlorinated pesticides (2,4-D and DDT) biodegradation at high concentrations using immobilized Pseudomonas fluorescens. AB - Degradation of two chlorinated pesticides (2,4-D and DDT) using a 54-mL glass column packed with tezontle (a low-cost basaltic scoria) was tested. Bacteria were cultured in YPG (yeast, peptone, and glucose) liquid medium at 32 degrees C. The rich medium was pumped during 24 h through the column to inoculate it. Later, the wasted medium was discharged and the pesticide added. Optical densities, TOC, and pesticide concentration were determined. Pesticide removals for 2,4-D (with initial concentration between 100 and 500 mg/L) were about 99%. DDT removal (at initial concentration of up to 150 mg/L) was as high as 55-99%. TOC removals for 2,4-D was in the 36-87% interval, whereas for DDT they were as high as 36-78%. PMID- 16047881 TI - Toxicity of the mosquito control pesticide Scourge to adult and larval grass shrimp (Palaemonetes pugio). AB - This study investigated the toxicity of various concentrations of technical resmethrin and Scourge on adult and larval Palaemonetes pugio, a common grass shrimp species. Two types of tests were conducted for each of the resmethrin formulations using adult and larval grass shrimp life stages, a 96-h static renewal aqueous test without sediment, and a 24-h static nonrenewal aqueous test with sediment. For resmethrin, the 96-h aqueous LC50 value for adult shrimp was 0.53 microg/L (95% confidence interval (CI): 0.46-0.60 microg/L), and for larval shrimp was 0.35 microg/L (95% CI: 0.28-0.42 microg/L). In the presence of sediment, technical resmethrin produced a 24-h LC50 value for adult shrimp of 5.44 microg/L (95% CI: 4.52-6.55 microg/L), and for larval shrimp of 2.15 microg/L (95% CI: 1.35-3.43 microg/L). For Scourge, the 96-h aqueous LC50 for adult shrimp was 2.08 microg/L (95% CI: 1.70-2.54 microg/L), and for larval shrimp was 0.36 microg/L (95% CI: 0.24-0.55 microg/L). The 24-h sediment test yielded an LC50 value of 16.12 microg/L (95% CI: 14.79-17.57 microg/L) for adult shrimp, and 14.16 microg/L (95% CI: 12.21-16.43 microg/L) for larvae. Adjusted LC50 values to reflect the 18% resmethrin concentration in Scourge are 0.37 microg/L (adult), 0.07 microg/L (larvae) for the 96-h aqueous test, and 2.90 microg/L (adult), 2.6 microg/L (larvae) for the 24-h sediment test. Larval grass shrimp were more sensitive to technical resmethrin and Scourge than the adult life stage. The results also demonstrate that synergized resmethrin is more toxic to P. pugio than the nonsynergized form, and that the presence of sediment decreases the toxicity of both resmethrin and Scourge. PMID- 16047882 TI - Application of an artificial sea salt solution to determine acute toxicity of herbicides to Proisotoma minuta (Collembola). AB - The acute toxicity test is described in this experiment where the Collembola species Proisotoma minuta was exposed to herbicides in an artificial sea salt solution for seven days. The salt solution did not prohibit the insects' reproduction system. The seven day LD50 values for trifluralin, pendimethalin, metolachlor, prometryn, paraquat, atrazine, fluometuron, and diuron were 3.48, 10.4, 12.4, 13.0, 23.1, 33.4, 250, and 711 mg L(-1), respectively. A good correlation between toxicity of the compounds and their lipophilicity and vapor pressure was recorded in this study. PMID- 16047883 TI - Effect of manure on glyphosate and trifluralin mineralization in soil. AB - Manure additions to soil may alter soil chemical, physical, and biological characteristics, and thereby change pesticide fate processes in soil. This is the first study to examine the impact of liquid hog manure amendments on glyphosate and trifluralin mineralization in soil. Experiments were conducted in soil microcosms in the laboratory for a total of 332 (glyphosate) and 430 (trifluralin) days. The rate and amount of mineralization of both glyphosate and trifluralin were significantly influenced by the additions of fresh manure to soil in the laboratory and by the history of manure applications in the field. However, the maximum difference in herbicide mineralization between soils that were free of manure application and those amended with manure in the field or in the laboratory was only 6.1% and 7.3% of that initially applied, for trifluralin and glyphosate, respectively. Therefore, we conclude that liquid hog manure application to soil will have no significant effect on the mineralization of glyphosate and trifluralin under field conditions. PMID- 16047884 TI - Natural products: seasonal variation in trichome counts and contents in Lycopersicum hirsutum f. glabratum. AB - Wild tomato leaves possess a pest-resistance mechanism in their glandular trichomes and the exudates they produce. Type IV and VI glandular trichomes on the leaves of five wild tomato accessions of Lycopersicum hirsutum f. glabratum (PI 126449, PI 134417, PI 134418, PI 251304, and LA 407) grown under greenhouse conditions were counted. Major chemical compounds from glandular leaf trichomes of the accessions tested were extracted, purified, and quantified at different periods during the growing seasons by gas chromatography (GC) and mass spectrometry (GC/MS). The toxicity of two methylketones (2-undecanone and 2 tridecanone), the major constituents of the accessions tested, to adults of the sweetpotato whitefly, Bemisia tabaci (Gennadius) and fourth instar larvae of the Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), was determined using no-choice bioassays. 2-undecanone caused 80% mortality of the fourth instar larvae of the CPB at the highest concentration tested (100 mg 2-undecanone mL(-1) of acetone) while 2-tridecanone caused 72% mortality of whiteflies at 20 mg 2 tridecanone mL(-1) of 65% ethanol. The concentration of 2-undecanone and 2 tridecanone on the leaves of each of the five accessions of L. hirsutum f. glabratum and the mass spectra of 2-undecanone and 2-tridecanone are presented. Overall, the concentration of 2-undecanone on the leaves was significantly higher than 2-tridecanone. Concentration of the two methylketones varied among accessions and sampling season. Plants produced the highest concentration of 2 undecanone during the month of August. On average, each plant of accession PI 251304 provided 354 g fresh leaves (averaging about 52,353 cm2 exposed leaf surface area) and produced 32.5 and 8.7 mg of 2-undecanone and 2-tridecanone, respectively. L. hirsutum f. glabratum accessions may become a valuable source of natural products, which could minimize reliance on synthetic pesticides. PMID- 16047885 TI - Evaluation of detection methods for genetically modified traits in genotypes resistant to European corn borer and herbicides. AB - Detection of genetically modified (GM) traits in corn (Zea mays L.) is urgently needed for preservation of genetic identity and marketing GM products. A laboratory study was conducted to evaluate the efficiency, accuracy, and reliability of different analytical methods to detect GM traits in corn. Samples with known fractions of GM concentrations (Bacillus thuringiensis [Bt], Liberty Link [LL] and stacked [Bt/LL] genes) from commercial seed companies and those derived from yellow kernels in a white corn ear, outcrossed by pollen of neighboring Bt hybrid, were tested by lateral flow quick test kits and by enzyme linked immunosorbent assay (ELISA)-based test strips purchased from different companies. Liberty Link hybrids are resistant to glufosinate (Liberty or Basta) herbicides, whereas Bt corn is developed for the control of European corn borer (Ostrinia nubilalis). Selected samples of GM concentrations were also tested in a commercial laboratory using DNA-based polymerase chain reaction (PCR) test. The results demonstrated that qualitative lateral flow quick tests could detect samples containing 1% or higher Bt and 2% or higher LL concentrations within the recommended time limit of the test. Faint test lines for samples containing 0.5 to 1% Bt or 1 to 2% LL concentrations appeared if samples remained in the test cup overnight. ELISA test strips detected the Bt content semiquantitatively in the range of 0.5 to 2.0%. Grain samples derived from non-Bt corn outcrossed by neighboring Bt pollen had usually lower GM concentrations than commercial GM seed samples. Both ELISA- and DNA-based PCR tests distinguished samples with GM concentrations between 0.1 to 0.5%, but the precision of quantification at this range was very low and results were highly inconsistent. PMID- 16047886 TI - Short chain fatty acids influence virulence properties of Salmonella enterica serovar Typhimurium. AB - The present study investigated SCFA exposed Salmonella enterica serovar Typhimurium for its virulence characteristics such as mouse lethality adhesion, surface hydrophobicity, phagocytic uptake, intracellular survival within the murine peritoneal macrophages, induction of acid tolerance response (ATR), and the expression of outer membrane proteins (OMPs). Lethal dose (LD50) for mice was found to be more for SCFA exposed cells as compared to the normal cells. In vitro adherence to murine intestinal epithelial cells as well as surface hydrophobicity of SCFA exposed cells were found to be lower than the normal cells. Though the phagocytic uptake of normal cells and SCFA exposed cells was similar, the SCFA exposed cells exhibited increased intracellular survival as compared to the normal cells, which were completely killed after 4 h. Moreover, SCFA exposed cells also survived exposure to extremely low pH (3.0). Outer membrane proteins extracted from SCFA exposed cells revealed an enhanced expression of proteins (porins) at 4% SCFA concentration. Thus, SCFA exposure may contribute to enhanced virulence of Salmonella enterica serovar Typhimurium by increased intracellular survival, induction of acid tolerance response, and the enhanced expression of outer membrane matrix proteins (porins). PMID- 16047887 TI - Vermicomposting of winery wastes: a laboratory study. AB - In Mediterranean countries, millions of tons of wastes from viticulture and winery industries are produced every year. This study describes the ability of the earthworm Eisenia andrei to compost different winery wastes (spent grape marc, vinasse biosolids, lees cakes, and vine shoots) into valuable agricultural products. The evolution of earthworm biomass and enzyme activities was tracked for 16 weeks of vermicomposting, on a laboratory scale. Increases in earthworm biomass for all winery wastes proved lower than in manure. Changes in hydrolytic enzymes and overall microbial activities during the vermicomposting process indicated the biodegradation of the winery wastes. Vermicomposting improved the agronomic value of the winery wastes by reducing the C:N ratio, conductivity and phytotoxicity, while increasing the humic materials, nutrient contents, and pH in all cases. Thus, winery wastes show potential as raw substrates in vermicomposting, although further research is needed to evaluate the feasibility of such wastes in large-scale vermicomposting systems. PMID- 16047888 TI - Effects of dietary zinc supplementation on hen performance, ammonia volatilization, and nitrogen retention in manure. AB - This investigation was undertaken to evaluate the effects of dietary ZnSO4 supplementation on ammonia volatilization and nitrogen retention in hen manure. One hundred twenty, 45-wk-old commercial Leghorn laying hens were sequentially fed diets with 1000, 2000, and 3000 ppm Zn as ZnSO4 (Zn-1000, Zn-2000, and Zn 3000), then followed by two control dietary periods with 114 ppm Zn (Control-1 and Control-2) for a total of five consecutive eight-day experiment periods, respectively. When hens were fed the 1000 and 2000 ppm Zn treatment diets, room ammonia levels were significantly reduced compared to the control diets. Dietary Zn treatments reduced the decomposition of uric acid, resulting in an increase in manure total-N retention compared to the control fed birds. The 1000 ppm Zn supplement had no adverse effects on hen body weight, feed consumption, egg production, egg weight, albumen height, or shell thickness. However, hens fed the diet containing 3000 ppm Zn had significantly depressed body weight, feed consumption, egg production, egg weight, and shell thickness. Zinc levels of egg contents increased linearly as dietary Zn levels increased. These levels in eggs would not be a problem for human consumption because these are much less than the daily Zn recommended dietary allowance. Although land application of such manure will not cause environmental problems or crop toxicity, proper monitoring of soil and crop Zn levels and effective nutrient management planning would be well advised. PMID- 16047889 TI - Adsorption/desorption of copper by a sandy soil amended with various rates of manure, sewage sludge, and incinerated sewage sludge. AB - Organic amendments are sometimes applied to agricultural soils to improve the physical, chemical, and microbiological properties of the soils. The organic fractions in these soil amendments also influence metal reaction, particularly the adsorption and desorption of metals, which, in turn, determine the bioavailability of the metals and hence their phytotoxicities. In this study, a Quincy fine sandy (mixed, mesic, Xeric Torripsamments) soil was treated with 0 to 160 g kg(-1) rates of either manure, sewage sludge (SS), or incinerated sewage sludge (ISS) and equilibrated in a greenhouse at near field capacity moisture content for 100 days. Following the incubation period, the soil was dried and adsorption of copper (Cu) was evaluated in a batch equilibration study at either 0, 100, 200, or 400 mg L(-1) Cu concentrations in a 0.01M CaCl2 solution. The desorption of adsorbed Cu was evaluated by three successive elutions in 0.01M CaCl2. Copper adsorption increased with an increase in manure rates. At the highest rate of manure addition (160 g kg(-1) soil), Cu adsorption was two-fold greater than that by the unamended soil at all rates of Cu additions. With increasing rates of Cu additions, the adsorption of Cu decreased from 99.4 to 77.6% of Cu applied to the 160 g kg(-1) manure amended soil. The desorption of Cu decreased with an increase in rate of manure amendment. Effects of sewage sludge amendments on Cu adsorption were somewhat similar to those as described for manure additions. Likewise, the desorption of Cu was the least at the high rate of SS addition (160 g kg(-1)), although at the lower rates there was not a clear indication of the rate effects. In contrast to the above two amendments, the ISS amendment had the least effect on Cu adsorption. At the highest rate of ISS amendment, the Cu adsorption was roughly 50% of that at the similar rate of either manure or SS amendments, across all Cu rates. PMID- 16047890 TI - Optimized condition for etching fused-silica phase gratings with inductively coupled plasma technology. AB - Polymer deposition is a serious problem associated with the etching of fused silica by use of inductively coupled plasma (ICP) technology, and it usually prevents further etching. We report an optimized etching condition under which no polymer deposition will occur for etching fused silica with ICP technology. Under the optimized etching condition, surfaces of the fabricated fused silica gratings are smooth and clean. Etch rate of fused silica is relatively high, and it demonstrates a linear relation between etched depth and working time. Results of the diffraction of gratings fabricated under the optimized etching condition match theoretical results well. PMID- 16047891 TI - Optimization of finite-length input volume holographic grating couplers illuminated by finite-width incident beams. AB - A finite volume holographic grating coupler (VHGC) normally illuminated with various incident-beam profiles (such as a Gaussian beam, a flat cosine-squared beam, and an exponential-decay beam) with finite beam widths for input coupling is rigorously analyzed by use of the finite-difference frequency-domain method. The effects of the incident-beam width, the incident-beam position, the incident beam profile, and the incident-beam angle of incidence on the input coupling efficiency are investigated. The optimum conditions for input coupling are determined. Both a VHGC embedded in the waveguide film region and a VHGC placed in the waveguide cover region are investigated. For a given finite VHGC, the input coupling efficiencies are strongly dependent on incident-beam widths, incident-beam positions, and incident-beam angles of incidence, but are only weakly dependent on incident-beam profiles. PMID- 16047892 TI - Angular sensitivities of volume gratings for substrate-mode optical interconnects. AB - The angular sensitivities of slanted volume gratings (VGs) illuminated by three dimensional (3-D) converging-diverging spherical Gaussian beams for substrate mode optical interconnects in microelectronics are analyzed by application of 3-D finite-beam rigorous coupled-wave analysis. Angular misalignments about the z, y, and x axes that correspond to yaw, pitch, and roll misalignments resulting from manufacturing tolerances of chips are investigated. Two cases of linear polarization of the central beam of the Gaussian are considered: E perpendicular K and H perpendicular K, where K is the grating vector. From worst-case manufacturing tolerances, the ranges of yaw, pitch, and roll misalignment angles are alpha = +/-1.17 degrees, beta= +/-3.04 degrees, and gamma = +/-3.04 degrees, respectively. Based on these ranges of misalignment angles, the decreases of diffraction efficiencies for slanted VGs that are due to both the yaw and the roll misalignments are relatively small. However, the efficiency of substrate mode optical interconnects achieved by slanted VGs could be reduced by 61.04% for E perpendicular K polarization and by 58.63% for H perpendicular K polarization because of the pitch misalignment. Thus the performance of a VG optical interconnect is most sensitive to pitch misalignment. PMID- 16047893 TI - Principles and design of multibeam interference devices: a microelectromechanical systems segment-deformable-mirror-based adaptive spectrum attenuator. AB - Fourier analysis of multibeam interference shows that the total electric field and relative time delay of the beams form a Fourier-transform pair. Fourier analysis-based multibeam interference analysis and device design is discussed in detail. The principle of the proposed segment-deformable-mirror-based adaptive spectrum attenuator is illustrated. PMID- 16047894 TI - Polarization microscopy by use of digital holography: application to optical fiber birefringence measurements. AB - We present a digital holographic microscope that permits one to image polarization state. This technique results from the coupling of digital holographic microscopy and polarization digital holography. The interference between two orthogonally polarized reference waves and the wave transmitted by a microscopic sample, magnified by a microscope objective, is recorded on a CCD camera. The off-axis geometry permits one to reconstruct separately from this single hologram two wavefronts that are used to image the object-wave Jones vector. We applied this technique to image the birefringence of a bent fiber. To evaluate the precision of the phase-difference measurement, the birefringence induced by internal stress in an optical fiber is measured and compared to the birefringence profile captured by a standard method, which had been developed to obtain high-resolution birefringence profiles of optical fibers. PMID- 16047895 TI - Relationship between microscanned image quality and fill factor of detectors. AB - Microscanning is an important technique in high-resolution electro-optical imaging. It can increase the resolution and improve the performance of imaging systems. For optimum design of a staring imaging system with microscanning modes it is necessary to choose the optimum microscanning mode according to the fill factor of the detector. Hence it is important to study the effect of the fill factor on the microscanning image quality. With some assumptions, we introduce the sampling-averaging modulation transfer function of a detector array at the spatial Nyquist frequency with which to study quantitatively the improvement in image quality of various microscanning modes for selected fill factors (1, 2/3, and 1/2). Analytical results show that the amount of improvement is closely associated with the fill factor. Finally, typical sampling imaging of focal plane arrays with these fill factors are simulated. Experimental results qualitatively describe the effect of the fill factor on the microscanning image and show good agreement with theoretical analysis. PMID- 16047896 TI - Moire fringe reduction by optical filters in integral three-dimensional imaging on a color flat-panel display. AB - We propose a method to reduce the color moire fringes that are attributable to the structure of a color flat-panel display in integral three-dimensional imaging. The method uses two types of optical low-pass filter, diffuser and defocus. The effectiveness of the method was confirmed in an experiment. We describe a way to design these filters with moire's residual energy and video signal energy as indices and demonstrate the validity of the model, which combines two filters to reduce moire fringes. PMID- 16047897 TI - Jones-matrix models for twisted-nematic liquid-crystal devices. AB - Twisted-nematic liquid-crystal devices having high spatial resolution are suitable for spatial light modulators. Phase-modulation characteristics of the devices have been widely studied, but the phase delay calculated using a conventional Jones-matrix model is slightly different from that measured using an interferometer. We propose a modified model whose matrix components are described by angular parameters that are related to the distribution of twist and tilt angles inside the liquid-crystal layer through differential equations. The model is used to simulate phase-modulation characteristics, and the result agrees well with the experimentally measured phase delay. PMID- 16047898 TI - Measurement of duty cycles of photoresist grating masks made on top of multilayer dielectric stacks. AB - We present a novel and nondestructive method for measuring the duty cycles (ratio of ridge width to period) of submicrometer rectangular photoresist gratings made on top of multilayer dielectric stacks. The method exploits the fact that the effective index of the leaky mode that has a strong evanescent tail in the cladding changes with the duty cycle of the grating situated at the interface between the top dielectric layer and the cladding. By comparing measured coupling angles of the leaky mode with a theoretical or experimentally calibrated relationship between coupling angles and duty cycle, one can determine the duty cycle of the grating. This method is applicable even when the grating period is less than the measurement wavelength. It is simple because it does not require any power measurement. Most importantly, it is virtually independent of groove depth. The physical principle of the method and the results of experimental verification are presented. PMID- 16047899 TI - Tensorial analysis of a Fourier-transform profilometric setup devoted to the evaluation of muscular contractions. AB - We explore the potential use of the Fourier-transform profilometry technique in in vivo studies of muscular contractions through the variation of muscle-group cross sections. Thanks to a tensorial analysis of the technique, a general expression of its sensitivity vector is established. It allows derivation of the expression of the resolution and the limit condition imposed by the spatial sampling of the fringe pattern. Key parameters that maximize the sensitivity are then simulated. A measurement system is accordingly built up and characterized. It is then successfully applied to the evaluation of the deformation of the forearm muscles during grasping exertions. PMID- 16047900 TI - Phase-shifting (Sagnac) interferometer with external phase control. AB - A novel ring configuration for phase-shifting interferometry with external phase shifting control is presented. The device is a polarization (ring) interferometer, in which the reference and test arms are parts of the same collimated beam. The key point is to manage the polarization of the light such that orthogonal linear polarizations describe counterpropagating paths in the ring interferometer. The phase shift between the two waves is externally controlled with a Pockels cell, which permits fast phase modulation without the need for moving parts inside the interferometer. PMID- 16047901 TI - Computational modeling of the propagation of light through liquid crystals containing twist disclinations based on the finite-difference time-domain method. AB - The finite-difference time-domain (FDTD) method is used to compute propagation of light through textured uniaxial nematic-liquid crystal (NLC) films containing various types of twist disclination (defect) lines. Computational modeling by the FDTD method provides an accurate prediction of the optical response in multidimensional and multiscale heterogeneities in NLC films in which significant spatial optic axis gradients are present. The computations based on the FDTD method are compared with those of the classic Berreman matrix-type method. As expected, significant deviations between predictions from the two methods are observed near the twist disclination line defects because lateral optic axis gradients are ignored in the matrix Berreman method. It is shown that the failure of Berreman's method to take into account lateral optic axis gradient effects leads to significant deviations in optical output. In addition, it is shown that the FDTD method is able to distinguish clearly different types of twist disclination lines. The FDTD optical simulation method can be used for understanding fundamental relationships between optical response and complex NLC defect textures in new liquid-crystal applications including liquid-crystal-based biosensors and rheo-optical characterization of flowing liquid crystals. PMID- 16047902 TI - Measurement of wave-front aberration in soft contact lenses by use of a Shack Hartmann wave-front sensor. AB - Lower- and higher-order wave-front aberrations of soft contact lenses were accurately measured with a Shack-Hartmann wave-front sensor. The soft contact lenses were placed in a wet cell filled with lens solution to prevent surface deformation and desiccation during measurements. Aberration measurements of conventional toric and multifocal soft contact lenses and a customized soft contact lens have proved that this method is reliable. A Shack-Hartmann wave front sensor can be used to assess optical quality of both conventional and customized soft contact lenses and to assist in enhancing lens quality control. PMID- 16047903 TI - Differences between real and predicted corneal shapes after aspherical corneal ablation. AB - We study the differences between real and expected corneal shapes, using an aspherical ablation algorithm with a known equation and avoiding the limitation imposed by most studies of refractive surgery in which the ablation equations are not known. We have calculated the theoretical corneal shape predicted by this algorithm, comparing this shape with the real corneal topography. The results indicate that the deviations that appear in the corneal shape are significant for visual performance and for the correction of eye aberrations. If we include in this analysis the effect of reflection losses and nonnormal incidence on the cornea, we can reduce corneal differences, but they will remain significant. These results confirm that it is essential to minimize corneal differences to achieve effective correction in refractive surgery. PMID- 16047904 TI - Analysis of clusters and uniformity of distribution of states of polarization on the Poincare sphere. AB - An analysis of the clusters and the uniformity of distribution of states of polarization on the surface of a Poincare sphere generated by rotating wave plates is given. The analysis of clusters of the states of polarization is based on a spherical radial distribution function. For uniform analysis of the distribution, two methods are proposed. The first method is based on calculation of the correlation coefficient; the second method is based on calculation of the angles between pairs of the states of polarization on the Poincare sphere. For polarization scramblers consisting of eight or more rotating wave plates, nonclustered and near-uniform distribution of states of polarization is obtained. PMID- 16047905 TI - Scattering and reflective properties of ordered mesoporous silica films. AB - Ordered mesoporous and nonporous silica films were studied by analysis of reflection spectra and hemispherical elastic light scattering (HELS). The real and imaginary parts of the index of the films were estimated from the reflection spectra. The HELS angular distribution of the mesoporous film shows a minimum, which has been interpreted as an interference pattern coming from the beams scattered by the mesopores. PMID- 16047906 TI - Rod and spherical silica microlenses fabricated by CO2 laser melting. AB - The fabrication and testing of glass microlenses with rod and spherical shapes are described. The sizes of the lenses range from tens of micrometers to several millimeters. The surfaces of the lenses were fabricated by the melting method. These surfaces have been studied by several methods. The theoretical behavior of the lenses was investigated by ray tracing. Some applications of the lenses are presented. PMID- 16047907 TI - Uncertainty analysis in the measurement of the spatial responsivity of infrared antennas. AB - The measurement of a two-dimensional spatial responsivity map of infrared antennas can be accomplished by use of an iterative deconvolution algorithm. The inputs of this algorithm are the spatial distribution of the laser beam irradiance illuminating the antenna-coupled detector and a map of the measured detector response as it moves through the illuminating beam. The beam irradiance distribution is obtained from knife-edge measurements of the beam waist region; this data set is fitted to a model of the beam. The uncertainties, errors, and artifacts of the measurement procedure are analyzed by principal-component analysis. This study has made it possible to refine the measurement protocol and to identify, classify, and filter undesirable sources of noise. The iterative deconvolution algorithm stops when a well-defined threshold is reached. Spatial maps of mean values and uncertainties have been obtained for the beam irradiance distribution, the scanned spatial response data, and the resultant spatial responsivity of the infrared antenna. Signal-to-noise ratios have been defined and compared, and the beam irradiance distribution characterization has been identified as the statistically weakest part of the measurement procedure. PMID- 16047908 TI - Threshold effects for resistance to optical damage and nonvolatile holographic storage properties in In:Mn:Fe:LiNbO3 crystals. AB - The threshold concentration for In2O3 was found in In:Mn:Fe:LiNbO3 crystals by measurement of the infrared spectra of the crystals. The resistance of the In:Mn:Fe:LiNbO3 crystals to optical damage is characterized by changes in photoinduced birefringence as well as by distortion of the transmitted beam pattern. The resistance increases remarkably when the concentration of In2O3 exceeds its threshold. The resistance to optical damage of a In(3.0 mol. %):Mn:Fe:LiNbO3 crystal is 2 orders of magnitude higher that of a Mn:Fe:LiNbO3 crystal. The dependence of defects on the resistance to optical damage of the In:Mn:Fe:LiNbO3 crystals is discussed in detail. Nonvolatile holographic storage was achieved for all crystals, and the sensitivity of the In(3.0 mol. %):Mn:Fe:LiNbO3 crystal is much higher than that of the others. PMID- 16047909 TI - Temporal statistics of the beam-wander contribution to scintillation in ground-to satellite optical links: an analytical approach. AB - The beam-wander contribution to the scintillation in a ground-to-satellite free space optical link is one of major importance. An analytical model, based on the duality between beam wander and angle-of-arrival fluctuations, is proposed for the temporal statistics. The expression of the probability density function of the log-amplitude fluctuations is first obtained. Then, the expressions of the spatial and temporal autocovariances are also obtained. We present plots of the beam-wander contribution to the log-amplitude variance, as a function of the transmitter aperture size and the turbulence accumulated in the propagation path. We also present the angular fluctuation and log-amplitude scintillation spectrum plots for some selected cases. PMID- 16047910 TI - Improved hollow-core photonic crystal fiber design for delivery of nanosecond pulses in laser micromachining applications. AB - We report the delivery of high-energy nanosecond pulses (approximately 65 ns pulse width) from a high-repetition-rate (up to 100 kHz) Q-switched Nd:YAG laser through the fundamental mode of a hollow-core photonic crystal fiber (HC-PCF) at 1064 nm. The guided mode in the HC-PCF has a low overlap with the glass, allowing delivery of pulses with energies above those attainable with other fibers. Energies greater than 0.5 mJ were delivered in a single spatial mode through the hollow-core fiber, providing the pulse energy and high beam quality required for micromachining of metals. Practical micromachining of a metal sheet by fiber delivery has been demonstrated. PMID- 16047911 TI - Realigning the communication paradigm in nursing case management. AB - Communication underlies all of case management (CM), and better comprehension of the processes of communication within the context of the CM role may facilitate positive patient and organizational outcomes, and assist providers and policy makers in designing effective CM systems. Thus, the purpose of this qualitative, pilot project was to explore dimensions of communication of the nurse case manager role within a hospital setting. The results describe a "hub and spoke" model of communications and suggest the utility of this familiar model for understanding the communication dynamics for those who work as nursing case managers in hospital settings. Findings suggest that nurse case managers are at various times both advocates and mediators--communications roles that may encompass somewhat contradictory demands and perspectives. PMID- 16047912 TI - Flexible client-driven in-home case management: an option to consider. AB - Changes in health services and care needs have created high demand for case management of in-home services. To address this challenge, several models of case management have been used. Evaluations to date suggest that clients need different approaches for different circumstances at different times to optimize cost-effectiveness. Accordingly, one Canadian home care program adopted flexible client-driven case management engaging clients as partners in flexibly selecting either an integrated team, consumer-managed or brokerage model of case management in keeping with their preferences and abilities. Using an exploratory, multimeasure quasi-experimental design, a generic model of program evaluation, and both quantitative and qualitative methods, researchers identified challenges in implementing this intervention, policy impediments the clients characteristically in each of the three case management models, and client, provider, and caregiver outcomes of flexible, client-driven care. While further longitudinal investigation is needed findings suggest several important considerations for those interested in this option for care management. Alternative case management models do attract different client groups, and having a choice does not alter care costs or outcomes. Flexible client-driven case management may be experienced positively by case managers and other providers. PMID- 16047913 TI - Community case management: a strategy to improve access to medical care in uninsured populations. AB - Case management is a rapidly growing strategy to help vulnerable populations access the health care they need in a fragmented U.S. service delivery system. A number of lessons can be learned from the successes and challenges of a developing, hospital-based, community-focused case management group in Sedgwick County, Kansas. The Community Case Management program has a case management team based at each of four hospitals. Each team has a social worker and a nurse, whose main goal is to help the uninsured find a primary health care home. This dynamic combination has several benefits. Each team member brings a vital skill set in helping the clients change behavior and access the care and support they need. Also, the nurse-social worker team concept creates a synergy in which team members support each other and create the best solution to their clients' needs. PMID- 16047914 TI - The journal of Jessie S. Sylvester. AB - An abridged version of a journal kept by an elderly woman offers a first-person casebook on issues of aging. Jessie S. Sylvester, a retired secretary who lived alone in Brooklyn, New York, kept a fastidious record of her daily activities between January 1976 and May 1978. She wrote unflinchingly of the challenges of old age: the loss of family and friends, the changes in her neighborhood, and the decline of her own faculties. She also chronicled the comfort she found in chores and errands, the joys of living in an urban village, and vital support provided at the local senior center. Introduction by Ellen Cassedy, who discovered and abridged the diary after Ms. Sylvester's death. Ms. Sylvester's journal is also available in the form of a one-woman play suitable for use at conferences; see Acknowledgments for contact information. PMID- 16047915 TI - Old and sleeping rough: elderly homeless persons on the streets of Boston. AB - Older individuals living on the streets of our urban cities are a unique sub group of the homeless population. No studies have been published about these elderly "rough sleepers" who face daunting obstacles to health care while facing a litany of health risks on the streets that are magnified by the physical and mental limitations of advancing years. To improve our understanding of this itinerant group, the Street Team of the Boston Health Care for the Homeless Program prospectively followed 30 individuals aged 60 or older living on Boston's streets for the four-year period from 2000 through 2003. This cohort included 8 (27%) women and 22 (73%) men ranging in age from 60 to 82 years. The average age was 67 years old At the end of the four year study period, 9 (30%) had died and 6 (20%) were in nursing homes. Despite intense efforts, only 5 (17%) found housing. Seven (23%) were still on Boston's streets after four years, and one was lost to follow-up. We conclude that elderly rough sleepers have high morbidity and mortality and pose significant challenges to programs seeking to provide housing and supportive health care services for this vulnerable sub-group of elderly homeless persons. New and creative housing options are needed, and the delicate issues of competency and guardianship must be addressed. PMID- 16047916 TI - Skin lesions in the elderly: precancer and cancer. AB - Skin lesions, both precancers and cancers, are more common among the elderly White population, especially in exposed body parts such as head face, neck, and hands. Earliest diagnosis is the key to treatment and survival. Awareness of a new skin lesion or change in a preexisting lesion, with or without pigmentation, flat or nodular, ulcerated or bleeding mandates prompt consultation followed by biopsy. For the cancer, surgical excision and repair remains the treatment of choice. Precancerous superficial lesions may be treated (after biopsy) with cryosurgery, topical chemotherapy, chemosurgery, or in cases of leukoplakia, electrodessication. Radiation may be considered for recurrent or metastatic disease. PMID- 16047917 TI - The Medicare Prescription Drug, Improvement and Modernization Act and the impact on care of people with chronic illness: benefit design and implementation issues. AB - The Medicare Prescription Drug Improvement and Modernization Act represents the most far-reaching and one of the most controversial reforms of the Medicare program since its enactment in 1965. The Act ushers in a new Medicare prescription drug benefit, revitalizes Medicare Advantage plans and promotes new approaches to care of Medicare beneficiaries with chronic illness. However, while the Act evidences a commitment to improving the care of beneficiaries with chronic illness, reform may prove elusive. The basic benefit design is daunting in its complexity andfor certain beneficiaries, coverage may be less generous and less comprehensive than currently available coverage. To ensure appropriate transition and treatment for dual eligibles and others with chronic illness, both Congress and the Department of Health and Human Services must be prepared to address a number of important benefit design and operations issues and be willing to make mid-course corrections and adjustment as the process unfolds. PMID- 16047918 TI - [Premature separation of the placenta--etiology and risk factors]. AB - OBJECTIVE: Placental abruption complicates approximately 1% of all pregnancies and remains a significant cause of both maternal and fetal morbidity and mortality. Several risk factors are associated with this complications, but a clear causal relation is diffucult to establish. THE AIM OF THE STUDY: The aim of the study was to determine and identify risk factors for placental abruption. DESIGN: Retrospective cohort study. SETTING: Dept. of Obstetrics and Gynaecology, Medical Faculty of Palacky University, Olomouc. SUBJECT AND METHOD: Retrospectively gathered cohort of 180 women was compared to 198 healthy women. We have focused on anthropometric, sociodemographic, behavioral, obstetric and fetal parameters. Statistical evaluation was done by Statsoft, Inc. (2001) Statistika CZ (Software system data analysis), version 6. RESULTS: The incidence of the placental abruption was 0.89%. We have confirmed the following risk factors as statistically significant (p values are given): Low-degree education (0.00002), prepregnancy weight of the mother (0.00), weight gain in pregnancy (0.00), higher parity (0.0272), smoking (0.03847), preeclampsia (0.00889), chorioamnionitis (0.00), premature rupture of membranes (0.00), recurrent spontaneous abortions (0.00), positive family history of deep venous thrombosis (0.0007) and intrauterine growth retardation (0.0386). CONCLUSION: Placental abruption has a large number of potential risk factors. The true biological cause and its etiopathogenic role is still to be elucidated. PMID- 16047919 TI - [Nalbuphine in obstetrical analgesia]. AB - OBJECTIVE: Comparison between particular methods of obstetrics analgesia, their analgetic efects and influence on the progress of delivery and postpartum adaptation of fetus. DESIGN OF THE STUDY: Prospective study. SETTING: The Department of Gynecology and Obstetrics of the Teaching Hospital and the 2nd Medical Faculty of the Charles University in Prague. METHODS: A prospective study of 92 patients who received intravenous infusion nalbuphine (28 women) or epidural analgesia (31 women) or pethidin (33 women). Assessment of maternal analgesia, satisaction was carried out. Apgar scores and resuscitative measures required for the neonate were note at delivery. RESULTS: The best analgesic effect was in the group with using peridural analgesia, where max. score was 3. The highest value was in the group of women who received pethidin. The most significant decrease of score after application of analgesia was in group with epidural analgesia, the least decrease of score was in the group with pethidin. The Apgar score at 1. minute was the lowest in the group with epidural analgesia, the highest score was in the group with nalbuphine. CONCLUSION: The mixed agonist/antagonist narcotic analgesic like nalbuphine have the place in clinical practice. The obstetric analgesia is very common in obstetrics practice. The epidural analgesia is very popular but not always accessible. Nalbuphine is the option which is effective in analgesia and have minimal side-effects. PMID- 16047920 TI - [Changes of coagulation parameters during stimulation in the IVF programme and in ovarian hyperstimulation syndrome]. AB - OBJECTIVE: Evaluation of changes in hemostasis system during the ovarian stimulation in IVF programme and in patients with severe form of ovarian hyperstimulation syndrome. DESIGN: Review article. SETTING: Unica, Institute for Reproductive Medicine, Brno. METHODS: Survey and discussion based on literature review. CONCLUSION: Activation of coagulation system during the ovarian stimulation and in the development of ovarian hyperstimulation syndrome is mentioned. The risk of thrombosis is increased if the patient is carrier of thrombophilic status. PMID- 16047921 TI - [Influence of exogenous supplementation with luteinizing hormone during controlled ovarian hyperstimulation on the results of IVF cycle]. AB - OBJECTIVE: To evaluate the effect of exogenous luteinizing hormone in in vitro fertilization cycles with depot GnRH agonist and low residual LH levels. DESIGN: Prospective randomized clinical trial. SETTING: 2nd Department of Gynecology and Obstetrics, Medical Faculty and L. Pasteur University Hospital Kosice METHODS: In the prospective randomized clinical trial the effect of exogenous LH on outcomes of assisted reproduction was evaluated. The total number of 68 in vitro fertilization cycles in normogonadotrophic women undergoing assisted reproduction with GnRH agonist down-regulation and recombinant FSH controlled ovarian stimulation were included. In all cycles the low residual LH level in the middle of stimulation was detected. The cycles were randomized into three groups. The first group was stimulated with pure recombinant FSH. In the second group the exogenous LH activity in the form of human menotrophin was added. And in the third group the human recombinant LH was added. RESULTS: Better outcomes of assisted reproduction were detected in both groups with exogenous LH activity. But these results, except the dosage of FSH, were not statistically significant. The increase of pregnancy rate by more than one fifth in these both groups can be considered as clinically relevant. CONCLUSIONS: It can not be positively proved at the base of our results that exogenous LH activity in cycles with low residual LH level can improve outcomes of assisted reproduction. It could be appropriate in the future to select women that can profit from exogenous LH activity according to other parameters not only to the LH serum level. PMID- 16047922 TI - [The map of superficial lymphatic system of the breast and relation to the sentinel lymph node]. AB - AIM: The aim of study was to topographically map the superficial lymphatic drainage of the breast and its relation to sentinel node. DESIGN: Anatomical study. SETTING: Department of Oncogynecology, Clinic of Obstetric and Gynecology of 1st Medical Faculty of Charles University and General Hospital, Prague. METHODS: The study was performed on 19 female cadavers. After slow intradermal administration of Patent Blau into the periareolar region, into the centers and into the borders of the individual quadrants of the breast, the lymphatics were visualized and a careful resection of these lymphatics and lymph nodes was performed. The lymphatics were drawn onto a schematic map of the breast on a transparent sheet and afterwards a summation of all schematic maps was performed. A map of the lymphatic vessels of the breast was based on this summation. RESULTS: After subcutaneous administration of patent blue were clearly visualized the subareolar plexus and draining sentinel node in the axilla, as well as one or two supplying lymphatics. Despite great variability in the lymphatic drainage of the breast, it was demonstrated that the natural dominant drainage for the outflow of lymph from the superficial areas of the breast are the axillary nodes. CONCLUSION: From the course of the lymphatic vessels it is evident that the breast cannot be divided into functionally separate segments. According to the lymphatic map of the breast, one can assume that in carcinoma of the breast it is the lymphatic drainage and not the localization of the tumor that plays a primary role in the development of metastases. One cannot ignore the existence of alternative drainage pathways of the breast, but their role in the beginning of dissemination of tumor cells in comparison with the axilla is negligible. Subdermal administration of Patent Blau is a reliable and effective method of visualizing lymphatic vessels and the draining lymph nodes of the breast. Due to the meandering course of lymphatics in the breast, we cannot understimate the role of radiation therapy after breast conserving surgery. PMID- 16047923 TI - [On the problem of sentinel lymphatic nodes examination in mammary cancer]. AB - OBJECTIVE: Evaluation of present approaches to the investigation of sentinel lymph nodes in breast cancer. DESIGN: A review article. SETTING: Obstetrics and Gynecology Department, Institute of Pathology and Department of Clinical Oncology of 1st Faculty of Medicine and General Teaching Hospital, Prague. SUBJECT: Sentinel lymph node biopsy can change surgical management of breast cancer and evaluation of regional lymph nodes. Sentinel nodes are reached first by the process of metastasis. Their status has been shown to rather accurately reflect the presence or absence of metastases in the axillary lymph nodes. The macrometastases (larger than 2 mm) are the most important ones. Step or serial sectioning and cytokeratins detection are necessary for findings of micrometastases (0.2-2 mm) or isolated tumor cells. Their prognostic significance is until now unclear and standard examination does not exist. CONCLUSION: Pathological examination of sentinel lymph nodes ought to identity all macrometastases (larger than 2 mm) as a minimum standard. Further investigations are necessary in order to evaluate the significance of micrometastases and isolated tumor cells and to find an optimal and standard processing. PMID- 16047924 TI - [Reconstruction procedures following pelvic exenterations]. AB - OBJECTIVE: Review of reconstruction procedures following pelvic exenterations. DESIGN: Review article. SETTING: Department of Obstetrics and Gynecology, Department of Urology, 1st Department of Surgery, Faculty Teaching Hospital and 1st Medical Faculty of the Charles University, Prague. METHODS: Review and critical assessment of published data. CONCLUSIONS: Reconstruction procedures are important part of pelvic exenterations. The procedures are crucial for following quality of life. Currently the most frequently used techniques for isolated pelvic floor support are omental flaps (carpets), for combined reconstruction of pelvic floor and vagina TRAM (transverse rectus abdominis musculocutaneus flap). Reconstructions prolong operation time; however they are accompanied with low morbidity and some techniques decrease total morbidity of exenterative procedure. Total and posterior exenterations require sigmoideostomy in vast majority of cases. Low rectal anastomosis might be used in cases of supralevator procedures. They cause high morbidity especially in patients following radiotherapy. In these patients temporary diverting colostomy is being recommended. A bowel segment is usually used for urinary diversion following total or anterior exenteration. Golden standard remain the incontinent ureteroenterostomies using ileum or colon transversum. Currently continent diversions are considered more often due to encouraging results and good quality of life. Heterotopic diversions, with continent conduit and cutaneous stoma, are frequently used. Risk of serious complications, especially fistulas and stoma stenosis, after all types of diversions is possible to reduce by using appropriate bowel segment not handicapped by previous radiotherapy. PMID- 16047925 TI - [Metastasing and relapsing "low grade" adenosquamous metaplastic breast cancer- is there a really indolent lesion? A description of three cases and review of literature]. AB - OBJECTIVE: To describe personnel experience with three unusual cases of low-grade adenosquamous carcinoma of the breast. Due to the aggressive course in two cases, the review of the literature does not concentrate on morphology and differential diagnosis only, but discuss overall biology of this lesion too. DESIGN: Three case reports. SETTING: Biopsy Lab s.r.o. and Sikl's Department of Pathology, Charles University and Faculty Hospital, Pilsen. METHODS: Included are cases from mammary register of Biopsy Lab s.r.o. and from personnel consulting registry of Prof. Michal. In details are documented both clinical presentations, including follow up, and spectrum of histologic patterns of the lesion. Immunohistochemistry performed by streptavidin-biotin system (LSAB+, Dako) as detection system and diaminobenzidin tetrahydrochlorid as chromogen, employed standard myoepitelial markers (actin, CD10, calponin and p63), hormonal receptors, oncogen Her-2/neu, CK7, CAM5.2, CD34 and CD117. RESULTS: Partial mastectomy and segmentectomy were performed in three women 46, 72 and 74 years old resp. for tumor, which size ranged from 20-35 mm in maximum diameter (mean 28 mm). Histology revealed low-grade adenosquamous metaplastic carcinoma displaying typical biphasic appearance combining regular tubular structures with surrounding storiform, sclerosing fibrous stroma. Variably present was characteristic squamous metaplasia of the gland forming either luminal morules or evident squamous pearls. Using immunohistochemistry, a total absence of myoepithelial layer in epithelial structures was confirmed. The stromal component expressed smooth muscle actin in two cases, one of which contained areas of heterologous chondrosseal differentiation. There were recognized metastases by one woman in two ipsilateral axillary lymph nodes mimicking benign breast heterotopia in one of them. One patient experienced recurrence in the scar 37 months after mastectomy where only mesenchymal heterologous component element was found. In two women with aggressive course the original biopsy was falsely interpreted, once as phyllodes tumor and secondly as benign sclerosing pseudotumor. CONCLUSION: low-grade adenosquamous carcinoma, despite its bland histology, should be understood as regular malignancy prone to frequent recurrences and recognized metastatic potential. It arises in the deep breast tissue and structurally resembles the microcystic adnexal carcinoma of the skin. Low-grade adenosquamous carcinoma, however, has nothing to do with syringomatous adenoma of the nipple, which is a benign tumor of the skin adnexa. Differential diagnosis includes spectrum of non-neoplastic slerosing lesions and above-mentioned phylloid tumor. The rarity of this neoplasm does not exclude deep knowledge. PMID- 16047926 TI - [Immune, inflammatory and hormonal changes in connection with laparoscopic surgery]. AB - OBJECTIVE: Analysis of the issue of laparoscopy on the intra and post-operative immune changes. SUBJECT: Review article. SETTING: Department of Obstetrics and Gynecology, Regional Hospital Kladno. SUBJECT AND METHOD: Review of published facts with regard to impact laparoscopic surgery on immune function. CONCLUSION: The impact of surgery on the intra and post-operative immune, hormonal and inflammatory response is significantly less after laparoscopy than with an open approach. PMID- 16047927 TI - [Changes in urethra mobility after TVT operation]. AB - OBJECTIVE: The aim of our study was to asses changes in the mobility of the whole urethra after successful TVT procedure. DESIGN: Prospective pilot study. SETTING: Obstet. Gynecol Department, General Teaching Hospital, 1st Medical Faculty, Charles University, Prague; EuroMISE Centre of the Charles University and Academy of Sciences, Prague, Czech Republic. METHODS: 101 women with proven stressed urinary incontinence were included in the study. After the TVT procedure 90 women were evaluated. As a part of the complex urogynecological investigation before surgery the transperineal ultrasound scan was performed in supine position, urinary bladder was filled to 300 ml. In the orthogonal system of coordinates the position and mobility of the whole urethra before surgery were assessed. Control examination was done 3-6 months after the surgery. The changes induced by the surgery were assessed. For the statistical evaluation t-test, Wilcox test, F test, Kruskal-Wallis test and ANOVA were used. RESULTS: Surgery significantly decreased the mobility of the whole parts of the urethra during maximal Valsalva, but the position at rest is not influenced. The women with high urethral mobility have high mobility after the surgery. The operation was more effect in patients with high mobility. Never the less the change of relative mobility is the same in all women. CONCLUSIONS: The information about the type of urethral mobility is important and may increase the success rate of TVT. Therefore the tension of the tape should be different for patients with different urethral mobility. PMID- 16047928 TI - [Laparoscopy in chronic pelvic pain--a prospective clinical study]. AB - OBJECTIVE: The aim of this study was to analyze the incidence, causes and management in women with chronic pelvic pain and to evaluate the role of laparoscopy. DESIGN: A prospective non-randomized clinical trial on 86 women with chronic pelvic pain. SETTING: Clinic of Gynecology and Obstetrics, JMF CU Martin, Slovak Republic. METHODS: A prospective clinical trial was performed on 86 patients with chronic pelvic pain, who have undergone laparoscopy from March 2003 to March 2004. Only patients with a pain history of at least 6 month were enrolled into this trial. Specific patient's history characteristics, laparoscopic and cytologic findings were reviewed and analyzed (pain interval, organic findings, preoperative ultrasound examination, previous surgical intervention, oral contraceptive usage, patient's medical history, menstrual cycle regularity, age, presence of dysmenorrhea). In all women, laparoscopy was performed under general anesthesia. RESULTS: During the study we have performed 309 diagnostic laparoscopic examinations, from which 86 (27.8%) were done due to chronic pelvic pain. The mean patient's age was 35.8 years (19-56). The mean parity was 1.6, ranging from 0-5. Pelvic organ pathology was present in 88.4% of the patients. The most frequent finding was endometriosis (31.4%). According to revised criteria of the American Fertility Society the presence of first, second, third and fourth stage of endometriosis was 55.6, 25.9, 11.1 and 7.4%, respectively. The most frequent occurrence of endometriotic lesions were on ligamenta sacrouterina (21.4%) and plica vesicouterina (19.0%). Pelvic adhesions, myomas, pelvic varicosities and chronic inflammatory process were present in 25.6, 15.1, 9.3 and 3.5% of the cases, respectively. No somatic origin of pain was identified at laparoscopy in 11.6% of patients. Preoperative ultrasonic examination with pelvic pathology findings were performed in 36 patients, and laparoscopy correlated with ultrasonographic findings in 31 (86.1%) cases. The average pain duration was 11.5 months (6-28) with the majority among women with history of previous surgical intervention (48.8%) and parturated women. Presence of pain was most common among women after 31 years of age. Predominantly, cytology examination of biological materials (peritoneal fluid, cyst fluid) revealed an increased histiocytic reaction in coincidence with chronic inflammation process in 31.6%. CONCLUSION: Invasive laparoscopy in chronic pelvic pain pertains to one of the most important examination procedures for its high specificity and sensitivity. Laparoscopy can reveal organic causes of pelvic pathology in 60% of cases with the possibility of following treatment. Our combined effort should stop the progression of such pathology leading to possible morphologic, functional and psychological alteration, especially among young women in fertile age. Today, endometriosis still remains the main cause of chronic pelvic pain in high percentage rate. PMID- 16047929 TI - ["Mixed" and "miscellaneous" vulvovaginitis: diagnostics and therapy of vaginal administration of nystatin and nifuratel]. AB - AIM OF THE STUDY: The evaluation of combined and miscellaneous vulvovaginal infections incidence and their treatment with combined vaginal products containing nifuratel and nystatin. DESIGN: Prospective study. SETTING: Gynecologic outpatient department LEVRET, Prague; Laboratories of Microbiology AescuLab, Prague. METHODS: 70 consecutive patients were examined with complaint of vaginal fluor and/or pruritus. We established macroscopic features of fluor, pH, amine test and mounts stained with Giemsa and Gram. We qualified the cases with more diagnostic criteria (mycosis, lactobacillosis, anaerobic vaginosis, aerobic vaginitis) as combined infection, those with no diagnostic criteria as miscellaneous. We treated all patients with vaginal tablets nystatin + nifuratel (Macmiror complex). We prescribed clotrimazol cream, if pruritus was present. We evaluated withdrawals of symptoms and relapses during 3 months after treatment. RESULTS: Combined infection was found in 21 patients from 70 (30%). The most frequent combination was that of mycosis and aerobic vaginitis (13/70, 18.6%) or mycosis and anaerobic vaginosis (4/70, 5.7%); 11 patients fulfilled criteria of no diagnosis. We concluded them as "miscelaneous". The treatment was successful in all cases, 10 women relapsed in 3 months. CONCLUSIONS: Combined vaginal infection findings are present very often (30%), likewise miscellaneous ones (15%) occur. The treatment of these women in successful with vaginal tablets with nystatin + nifuratel. PMID- 16047930 TI - [Fistula as a complication of pelvic actinomycosis--two case reports]. AB - OBJECTIVE: The presentation of two unusual cases of pelvic actinomycosis. SUBJECT: Case reports. SETTINGS: Department of Obstetrics and Gynecology, Hospital Kladno. SUBJECT AND METHOD: The observation of two cases of pelvic actinomycosis. These cases were complicated by the abdominal wall fistula in one case and vaginal fistula in the other. CONCLUSION: Actinomycosis is a chronic disease with tendency to progress per continuitatem into neighbouring tissues and with tendency to formation of fistulas. Two less common localisations of progress of the disease are described; it means the abdominal wall and vagina. In spite of a more advanced stadium, if adequately treated, a recovery ad integrum is common. PMID- 16047931 TI - Global Advisory Committee on Vaccine Safety, 9-10 June 2005. PMID- 16047932 TI - Do multiple outcomes and conditional factors improve prediction of batterer reassault? AB - Researchers and practitioners recognize the need to develop better prediction of abuse and reassault among men referred to batterer programs. Previous approaches have produced relatively weak predictions for primarily dichotomized "reassault versus no reassault" outcomes. The primary objective of this research was to improve prediction using multinomial logistic regressions with multiple outcomes and conditional factors. We used a unique data set from a multisite batterer program evaluation with a large sample size (n = 840), a longitudinal 15-month follow-up, and high response rates (70% of the women). Multiple outcomes included no abuse, verbal abuse or controlling behavior, threats, one-time reassault, and repeated reassaults. Using multiple outcomes improved prediction with risk markers over prediction of a dichotomous outcome and suggests that "repeat reassault" should be distinguished from "one-time reassault." Conditional factors did not substantially improve prediction. The low "correct classification" suggests, however, the difficulty of identifying the most dangerous batterers. PMID- 16047933 TI - A representative measure of psychological aggression and its severity. AB - Surveys of psychological aggression have been plagued by a variety of conceptual and measurement problems. A new measure was devised to more systematically cover a full range of psychologically aggressive actions; to include items for each dimension/category at a milder, moderate, and severe level; to be applicable to dating as well as marital relationships; and to utilize items that were broader in nature to capture all instances of particular types of psychological aggression. Three hundred and eighty-three college students rated the 51 items as to their degree of "psychological abusiveness." The 17 categories were rated as mostly distinct from each other; almost every category had statistically distinct mild, moderate and severe items; the overall scale yielded basically normal psychometric properties; and the total score, as well as the scores for the mild, moderate, and severe items, all had very high internal consistency. Ratings of these items appear to be distinct from social desirability as well as from a number of attitudinal response sets, and only sex of the participant was significantly, although weakly, correlated with overall ratings of the psychological aggression items. This measure may provide for more systematic investigation into the concept of psychological aggression. PMID- 16047934 TI - Child neglect and adolescent violence: examining the effects of self-control and peer rejection. AB - Child maltreatment researchers have often suggested that experiences with child neglect have long-term, negative effects. Child neglect is thought to have particularly adverse effects on self-control, peer relations, and delinquency. In this research, we examine the relationship of child neglect with adolescent violence via self-control and peer rejection. Using prospective, longitudinal data from a community sample, we find that child neglect adversely affects peer rejection and violence. Neglected children were more likely to be rejected by their peers in early adolescence and were more likely to be violent later in adolescence. Contrary to theoretical predictions, child neglect was not a significant predictor of self-control. Implications for delinquency and child maltreatment researchers are discussed. PMID- 16047935 TI - What really happened? A validation study of rape survivors' help-seeking experiences with the legal and medical systems. AB - Much of what is known about rape survivors' experiences with the legal and medical systems has come from victims' accounts; rarely have researchers collected "the other side of the story" to find out what system personnel say did or did not happen in these interactions. In the current study, rape survivors who sought emergency medical care were interviewed before their hospital discharge about what services they received and how they were treated by social system personnel. Corresponding accounts were then collected from doctors, nurses, and police officers. There was significant interrater reliability between the survivors and legal and medical system personnel regarding what services were or were not provided ("service delivery") and if system personnel engaged in "secondary victimization" behaviors (i.e., statements/actions that could be distressing to victims). However, police officers and doctors significantly underestimated the impact they were having on survivors. Victims reported significantly more post-system-contact distress than service providers thought they were experiencing. PMID- 16047936 TI - Police-reported intimate partner violence during pregnancy: who is at risk? AB - The objective of this retrospective cohort study was to determine risk factors for police-reported intimate partner violence (IPV) during pregnancy among Seattle residents with a registered live birth or fetal death in Washington State. Logistic regression was used to calculate adjusted odds ratios (aOR) and 95% confidence intervals for the association between demographic, behavioral, and obstetric history risk factors and any, physical, and non-physical police reported IPV. Significant risk factors for any police-reported IPV during pregnancy included unmarried status (aOR 2.36), public health program use (aOR 1.33), smoking or alcohol use during pregnancy (aORs 1.45 and 1.80, respectively), previous live birth (aOR 1.39), and previous spontaneous or induced abortion (aORs 1.39 and 1.34, respectively). Risk factors for physical IPV varied only slightly from those for any IPV, and fewer factors were associated with nonphysical IPV. Demographic, behavioral, and obstetric history risk factors are potential markers of IPV risk during pregnancy. PMID- 16047937 TI - Association between physical partner violence, posttraumatic stress, childhood trauma, and suicide attempts in a community sample of women. AB - Our objective was to estimate the prevalence of intimate partner violence (IPV) and to explore its association with childhood maltreatment, substance misuse, posttraumatic stress, and suicidal behavior in a representative community sample of women. IPV was operationalized as a "physical attack or beating by a spouse, boyfriend, or live-in partner." We surveyed 637 women in Memphis, Tennessee, by telephone survey. Sixteen percent reported ever experiencing IPV by a male partner, and 75% endorsed multiple assaultive acts. Of abused women, 5.9% met current PTSD diagnostic criteria, and an additional 11.8% were assessed with subthreshold symptoms. Abused women were more likely than other women to be divorced, to have less than 13 years education, to endorse high levels of childhood victimization, to have abused drugs and alcohol, and to have attempted suicide. Twenty-three percent of IPV+ (abused) women reported a suicide attempt at some time in their lives compared with 3% of IPV- (nonabused) women (p < .0001). Further, multiple logistic regression analysis showed that childhood sexual and emotional abuse and low educational attainment were the only significant predictors of IPV. These results suggest that in women who endorse IPV, careful inquiry of past abuse, trauma-related symptoms, suicidal behavior, and drug use may be important, so that interventions can be both timely and appropriate. PMID- 16047938 TI - Assessing physical, sexual, and psychological violence perpetrated by intimate male partners toward women: a Spanish cross-sectional study. AB - There have been many studies on the impact of intimate partner violence (IPV) on women's health, there being agreement on its detrimental effect. Research has focused mainly on the impact of physical violence on health, with few studies assessing the effect of sexual and psychological violence. Furthermore, there are many differences in the way violence experienced by women is assessed. While some researchers use available instruments, others develop their own questionnaires. This article gives detailed information about physical, sexual, and psychological violence, lifetime history of women's victimization, and aspects of women's behavior and feelings obtained with the questionnaire used in a Spanish cross sectional study. Our results corroborate that IPV is not homogeneous, it being necessary to ask women about each type of violence they have experienced. Furthermore, to accurately assess the impact of IPV on women's health, it is necessary to control for other variables that also have detrimental effects on health. PMID- 16047939 TI - [Treatment of retinal branch vein occlusion]. PMID- 16047940 TI - [Characteristics of N-methyl-N-nitrosourea-induced retinal degeneration in animals and application for the therapy of human retinitis pigmentosa]. AB - BACKGROUND: Retinitis pigmentosa(RP) is a human disease characterized by loss of photoreceptor cells, especially rods, leading to visual disturbance and eventually to blindness. Effective treatment for RP control is still unavailable. The establishment of reliable animal models is essential for a better understanding of this disease, and for the development of therapeutic intervention. Here we summarize the establishment of N-methyl-N-nitrosourea (MNU) induced retinal degeneration in animals, and success in disease control using this model. RESULTS: Retinal damage induced by MNU was highly reproducible and involved photoreceptor cell loss. It was obvious in all animals at approximately 7 days following a single systemic administration of MNU to adult mice (60 mg/kg), rats (60-75 mg/kg), hamsters (90 mg/kg), shrews (65 mg/kg), and monkeys (40 mg/kg). Extensive investigation in the rats revealed that MNU-induced photoreceptor cell loss was due to apoptosis with a decrease of Bcl-2 protein, increase of Bax protein, and activation of caspase families. Therapeutic to control MNU-induced photoreceptor cell loss in rats was evaluated with caspase-3 inhibitor (Ac-DEVD-CHO), nicotinamide(NAM), and docosahexaenoic acid(DHA); 4,000ng Ac-DEVD-CHO injected intravitreally 0 and 10 h after MNU suppressed disease progression, 25-1,000 mg/kg NAM subcutaneously injected concurrently or subsequently to MNU reversed retinal damage, and dietary supplementation of 9.5% DHA counteracted photoreceptor cell loss. CONCLUSION: Although the mechanisms triggering pathogenesis and the apoptotic cascade may differ between animals and humans, MNU-induced retinal degeneration is caused by photoreceptor cell apoptosis. Thus, suppression of MNU-induced photoreceptor cell apoptosis in animals may provide therapeutic information for RP control in humans. PMID- 16047941 TI - [Molecular mechanism of diabetic retinopathy: role of advanced glycation end products(AGEs) and their receptor (RAGE) in the pathogenesis of diabetic retinopathy]. AB - There is a growing body of evidence that advanced glycation end product receptor(AGE-RAGE) interaction elicits oxidative stress generation, thus indicating that it is involved in the pathogenesis of diabetic retinopathy. Inhibition of AGE formation or blockade of the downstream RAGE signaling is a promising therapeutic strategy for treatment of patients with diabetic retinopathy. PMID- 16047942 TI - [Cellular characteristics of cultured non-pigmented ciliary epithelium from adult pigs]. AB - PURPOSE: To compare the reactive proliferation of non-pigmented ciliary epithelial cells in patients with cyclitis, or after cyclophotocoagulation and cyclocryocoagulation, and the cultured non-pigmented ciliary epithelial cells from adult pigs. METHODS: Porcine ciliary epithelial cells were cultured and non pigmented ciliary epithelial cells were isolated. Detection of DNA synthesis, morphological observation by a phase contrast microscope and a transmission electron microscope, and staining of senescence-associated beta-galactosidase were carried out. RESULTS: The cells proliferated without showing contact inhibition of growth or reconstitution of epithelial morphology. With a decrease of proliferative activity, the cultured cells expressed senescence-associated beta-galactosidase. Although DNA synthesis persisted for a long time, some cells in later culture periods showed morphologically abnormal nuclei or plural nuclei indicating dysfunction of cell division, or apoptotic features. CONCLUSION: The uncontrolled growth and loss of the epithelial nature of non-pigmented ciliary epithelial cells in vitro resembles the process of formation of cyclitic membrane and proliferation of ciliary epithelium after cyclophotocoagulation and cyclocryocoagulation in patients. Observatin of the behavior of cultured non pigment epithelial cells could aid in understanding the mechanism of cyclitic membrane formation. PMID- 16047943 TI - [Clinical interpretation of the subjective complaint of dryness in contact lens wearers]. AB - PURPOSE: Patients wearing contact lenses who have the feeling of dryness without any objective symptoms of dry eye are sometimes seen in clinics. The purpose of this study was to determine if there was a difference in the clinical signs, personality, and mental condition between a group of contact-lens wearers with no symptoms and a group of contact-lens wearers with subjective symptoms of the feeling of dryness. SUBJECTS AND METHODS: Eighty-two patients who had no trouble with wearing soft contact lenses (SCL) were employed in this study. The patients were asked to fill out a questionnaire and Yatabe Guilford Personality test (Y-G test), and the height of tear meniscus and fluorescein staining were measured. RESULTS: Fifty-three patients (65%) complained of a feeling of dryness. There were no significant differences between the presence of the feeling of dryness and types of SCL, the height of tear meniscus, and the severity of fluorescein staining. On the other hand, there was a significant increase in the scores of positive, subjective, and impulsive personality trais on the Y-G test in the symptomatic group. CONCLUSION: For patients who have subjective symptoms of the feeling of dryness, careful observation of objective symptoms should be carried out, and their personality and mental condition should also be evaluated. PMID- 16047944 TI - [Serous macular detachment combined with branch retinal vein occlusion]. AB - PURPOSE: To report frequency, clinical characteristics, treatment, and the complications of branch retinal vein occlusion (BRVO) with serous macular detachment. PATIENTS AND METHODS: We retrospectively studied 22 eyes of 22 patients in 111 eyes with acute BRVO, whose eyes had serous macular detachment that was detected by optical coherence tomography (OCT). Fluorescein angiography was conducted in 19 of the 22 eyes. Fourteen of the 22 eyes underwent scatter laser photocoagulation of the BRVO area. We observed serial OCT findings before and after treatment. RESULTS: Approximately 20% of the BRVO eyes had serous macular detachment. The superotemporal vein was occluded in 15 eyes and the inferotemporal vein was involved in 7 eyes. Four eyes were ischemic and 15 eyes were not ischemic. Extensive dye leakage was observed in the BRVO area in all examined eyes (19 eyes). The occlusion area of perifoveal capillary network ranged from 5 to 60%, with an average of 40%. OCT demonstrated pure serous macular detachment in 13 eyes and the remaining 9 eyes had both serous macular detachment and cystoid macular edema(CME). The occlusion area of perifoveal capillaries in these 9 eyes was more than 20%. Serous macular detachment was resolved in 11 of 14 eyes (80%) 6 months after laser treatment. The average period for resolution of macular detachment was 3.4 months after treatment. Visual acuity was improved more than 2 lines in 8 of the treated 11 eyes (73%). Hard exudates appeared in the posterior fundus in 13 of 14 treated eyes (93%) and in 16 of the total of 22 eyes (73%) in the follow-up period. Massive macular hard exudates and ensuing macular atrophy resulted in poor visual outcome. CONCLUSIONS: Serous macular detachment is one of the patterns of macular edema in BRVO. OCT is an essential tool to detect it. Leakage from the entire BRVO area seems to travel via subretinal space by gravity or other factors and may form serous detachment in the macular area. Laser photocoagulation is indicated for early resolution of serous macular detachment. The major complication of serous detachment is the deposit of macular hard exudates, which may result in poor visual outcome. PMID- 16047945 TI - Aspects of oxalosis associated with aspergillosis in pathology specimens. AB - Oxalosis (calcium oxalate deposition) is associated with various conditions, including aspergillosis. Some Aspergillus species produce oxalic acid, which reacts with blood or tissue calcium to precipitate calcium oxalate. Calcium oxalate crystals exhibit various shapes and are strongly birefringent. These occur in cytological specimens, as well as in tissues of patients with Aspergillus infection. Aspergillus species are hyaline septate moulds, and they can be accurately recognized in pathology specimens only if conidial heads (fruiting heads) are present. When these structures are not observed, detection of associated oxalosis in a mould infection supports the pathological diagnosis of aspergillosis. The presence of oxalosis is helpful when microbiological identification or immunohistological techniques for fungi are not available. Calcium oxalate crystals can induce cellular injury by several mechanisms, and there is increasing evidence that oxalosis-induced tissue damage may occasionally lead to a poor clinical outcome. This review discusses the diagnostic value and the potential clinical significance of oxalosis associated with aspergillosis. PMID- 16047946 TI - Histiocyte-like cells expressing factor XIIIa do not belong to the neoplastic cell population in malignant fibrous histiocytoma. AB - The term malignant fibrous histiocytoma (MFH) is widely used for pleomorphic soft tissue sarcomas without a specific line of differentiation. MFH is included in the category of fibrohistiocytic soft tissue tumors. MFH has a broad range of histological appearances, and it has several subtypes. All of these subtypes are composed of spindled fibroblast-like cells, undifferentiated cells, and histiocytic or histiocyte-like cells. A large number of fibroblast-like and pleomorphic cells express factor XIIIa in MFH. The cytological pleomorphism of factor XIIIa cells suggests that these cells may belong to the neoplastic population. It is equally possible that the factor XIIIa-positive cells are only activated stromal cells. The relation of factor XIIIa-positive cells to the neoplastic cell population in MFH is addressed in the present study. A morphometric approach compares the measure of nuclear pleomorphism of the factor XIIIa-positive cells with that of the factor XIIIa-negative tumor cells in high grade MFH. The immunohistochemical approach compares the factor XIIIa-positive and -negative cell populations with regard to mutations of p53 tumor suppressor gene in p53-positive MFH cases. We selected 58 cases of soft tissue pleomorphic or storiform-pleomorphic MFH on the basis of histopathological examinations. A combination of incident light immunofluorescence for factor XIIIa and transmitted light examination for nuclear staining was used for morphometrical analysis. We found cytoplasmic factor XIIIa positivity in at least 2% of cells in 39 cases; the number of factor XIIIa-positive cells was under 0.5% in two cases, and the number of factor-positive cells ranged between 0.5% and 2% in 13 cases. Eighteen cases were analyzed with nuclear morphometry. We found that mean nuclear area and mean nuclear Ferret diameter in factor XIIIa-positive cells differed significantly from those of the tumor cells in all cases. The mean nuclear roundness factor differed significantly only in four cases. The latter finding showed that the microscopic polymorphism of factor XIIIa cells is measurable and is not merely a suspicion. The immunohistochemical positivity for p53 positivity can be accepted as the manifestation of a missense mutation of TP53 gene and as a marker of neoplastic cells. The simultaneous immunohistochemical detection of factor XIIIa and p53 in the same section revealed that factor XIIIa-positive cells were invariably p53 negative in MFH. This finding implies that the factor XIIIa cell population is non-neoplastic and belongs to the stromal component of MFH. PMID- 16047947 TI - Expression of CD163 in the liver of patients with viral hepatitis. AB - CD163 is a marker of activated macrophages, and increased levels of soluble CD163 have been detected in sera obtained from patients with hepatitis. The aim of this study was to detect the expression of CD163 in the liver from patients with viral hepatitis. Frozen sections of liver specimens were obtained from 5 patients with acute viral hepatitis (AH) and from 23 patients with chronic viral hepatitis (CH). The expression of CD163 in the liver was determined immunohistochemically using monoclonal antibody to human CD163. Double immunostaining was done to assess those cell types that express CD163 in the liver. The frequencies of CD163 positive cells were significantly higher both in the portal areas and in the hepatic lobules in the liver of patients with AH compared to those with CH (p < 0.05). Double immunostaining revealed that most of the CD163-positive cells were macrophages and Kupffer cells, because they expressed CD68. The expression of CD163 was very low in endothelial cells and liver stellate cells. This study shows that macrophages are activated in hepatitis liver. PMID- 16047948 TI - Hybrid chromophobe renal cell neoplasm. AB - Hybrid renal cell neoplasms (HRCNs) containing areas of tumor cells displaying cytological features of chromophobe renal cell carcinoma (CHRCC) and renal oncocytoma (RO) have been recently described in patients with renal oncocytosis and Birt-Hogg-Dube (BHD) syndrome (autosomal dominant genodermatosis). In this study, we identified cases of sporadic HRCN. We reviewed 425 consecutive renal cell carcinomas (RCC), 18 CHRCC, six HRCN, and 25 RO. Five HRCN were identified, including four from the group of RCC and two from RO. Patient age ranged from 40 to 68 years (mean age: 54 years), and the male:female ratio was 4:1. Tumors measured from 1.8 to 5 cm (mean diameter: 3.0 cm). Tumoral necrosis was not seen. Vascular invasion into medium-sized veins was identified in one HRCN. Chromophobe cells accounted for 20-80% of the tumors. Hale's colloidal stain showed weak to moderate diffuse cytoplasmic staining in scattered cells corresponding to those displaying routine staining features of chromophobe cells. Areas of oncocytic cells in studied tumors and control oncocytomas showed negative or focal cytoplasmic staining usually bordering extra- or intra-cytoplasmic lumina. Immunostaining for cytokeratin 7 and vimentin showed focal immunoreactivity in three cases and negative reactivity in all six cases, respectively. None of the study cases had microscopic RO, as commonly seen in renal oncocytosis, or were associated with BHD syndrome Sporadic HRCN accounted for 1% of RCC. They were of smaller size than RCC and were associated with a favorable prognosis. PMID- 16047949 TI - Effects of adrenergic agents on rat peritoneal macrophages activated in vitro by acetylated low-density lipoprotein. AB - Macrophages interact with modified lipoproteins and alter their functional status. In this study, the effects of the adrenergic agents adrenaline, isoproterenol, and dobutamine on macrophages activated with acetylated low density lipoprotein were tested. The aim of this investigation was to determine whether adrenergic agents influence biologically significant functions of these cells in an in vitro model of macrophage-lipoprotein acute interaction. Rat peritoneal macrophages were incubated with acetylated low-density lipoprotein for 16 h, with or without adrenergic agents. Hydrogen peroxide and nitric oxide production and acid phosphatase activities in the supernatant and cell lysate were assayed. Adrenaline and isoproterenol inhibited the production of hydrogen peroxide, stimulated the production of nitric oxide, and increased the extracellular activity of acid phosphatase in the lipoprotein-activated cells. Dobutamine increased the extracellular, but decreased the intracellular acid phosphatase activity. Adrenaline and dobutamine also produced changes in the cell size and nuclear morphology of the macrophages. Macrophages are activated in vitro by acetylated low-density lipoprotein, and their functions and morphology are modified under the influence of adrenergic agents. Certain effects could be attributed to changes in cyclic AMP levels. PMID- 16047950 TI - Proliferation and apoptosis in the corneal stroma in longterm follow-up after photorefractive keratectomy. AB - This study aimed at investigating the proliferation and apoptosis of corneal cells following photorefractive keratectomy (PRK) treatment. PRK (-6.0 D correction) was performed with the Asclepion-Meditec MEL70 G-scan excimer laser on the right eye of each of 33 rabbits under combined local and general anaesthesia. Animals were sacrificed at 4 h, 1, 4, 7, 14, 28, 56, and 112 days postoperatively, and corneal samples from these eight groups were examined histologically. Stromal cell proliferation was evaluated by immunocytochemical analysis of Ki67. Apoptosis was detected using the terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling (TUNEL) assay method. The untreated left eyes served as controls. Ki67 positivity was detected in the upper stroma on day 1, 4, 7, and 14, and keratocyte apoptosis on day 1, 4, 7, and 14 after PRK, but not at an earlier or later time. Neither Ki67 positivity nor apoptotic activity was observed in the controls (untreated corneas). PRK was found to trigger proliferation and apoptosis of corneal keratocytes. The frequency and spatial distribution of keratocyte proliferation and apoptosis are likely to be important determinants of the corneal wound healing process, but the detailed regulatory mechanisms have not yet been characterized. PMID- 16047951 TI - Syringomatous adenoma of the nipple: a case report. AB - We present a case of a 33-year-old woman who underwent excisional breast biopsy due to a left nipple mass. Histological examination revealed the morphologic and immunohistochemical pattern of syringomatous adenoma of the nipple. This is a rare lesion of the breast that can clinically mimic breast carcinoma, but harbors a benign and only locally aggressive course. Awareness of both the clinician and the pathologist for the possibility of diagnosing this tumor in the nipple region is mandatory to avoid mastectomy and lymph node dissection. PMID- 16047952 TI - Malignant struma ovarii: a case report. AB - Malignant struma ovarii is a rare form of the ovarian germ cell tumors. Hence, diagnosis and management of malignant struma ovarii have not been clearly defined. We present the case of a 34-year-old woman with papillary carcinoma arising in struma ovarii. The malignant component of this tumor was detected after laparoscopic removal, and a re-staging operation was performed afterwards. There was no evidence of clinical malignancy or metastases. In this paper, clinical features, treatment guidelines, diagnostic features, and immunohistochemical characteristics of this tumor are reviewed. PMID- 16047953 TI - The independent nurse role in community care gerontological nursing: special series-part 2. PMID- 16047954 TI - Feeding beliefs of certified nurse assistants in the nursing home: a factor influencing practice. AB - In this study, the author examined the feeding beliefs of 20 certified nurse assistants (CNAs) working in nursing homes using Q methodology and semistructured interviews. Beliefs are defined as a combination of CNA feeding knowledge, experience, and values. Two groups of CNAs with contra belief systems emerged from the analysis. "Social feeders" believe feeding is a time to socialize with residents and "technical feeders" believe prroviding adequate nutrition is the main goal when feeding. CNAs felt their beliefs influenced their feeding practices. Training programs need to include all factors that may influence CNA feeding practices, such as CNAs' beliefs, resident characteristics, and institutional factors. PMID- 16047955 TI - The oral health of older adults: an interdisciplinary mandate. PMID- 16047956 TI - Community care gerontological nursing: the independent nurse's role. AB - This focused ethnographic study examined the perspectives of seven nurses developing independent practices serving community-based elderly individuals. Six of the nurses' clients, five of whom were foster care providers, were also interviewed. The nursing activities described by these nurses included assessment, documentation, teaching, delegation and assignment, monitoring, advocacy, and support. The services valued by the clients included the nurses' problem-solving approach, advocacy with physicians, and assistance in meeting state licensure requirements. The clients also identified the following benefits: support in providing care to their residents, continuity of care, and personal relationships they developed with the nurses. PMID- 16047957 TI - The evolution of hospice in America: nursing's role in the movement. AB - In the current society, many individuals fear death and the feelings of suffering and loneliness that often accompany death. Two visionaries in the United States, Florence Wald and Dr. Elisabeth Kubler-Ross, recognized these fears and planned the nation's first hospice movement in the 1970s. The hospice philosophy continues to prosper in the new millennium. In this article, the founding American hospice's philosophy, types of facilities, standards, health team composition, patient demographics, organizations, reimbursement, and research are compared and contrasted with those of the current hospice movement. Existing issues with the modern movement are also discussed. PMID- 16047958 TI - Managing fecal incontinence: self-care practices of older adults. AB - Little is known about the ways in which community-dwelling elderly individuals manage fecal incontinence (FI) in their daily lives. In this study, community dwelling elderly individuals were surveyed at clinics of a health maintenance organization (HMO) to describe the self-care practices used to manage FI and to examine factors that influenced the number of self-care practices used and the reporting of FI to a health care practitioner. Responses of 242 elderly individuals who reported that they had FI several times per year were analyzed. The self-care practices used most commonly were changing diet, wearing a sanitary pad/brief, and reducing activity or exercise. Elderly women and those with a greater severity of FI and more chronic health problems engaged in more self-care practices. Factors associated with reporting FI to a clinician were considering FI to be a problem, uncertainty about the cause of FI, and changing diet to avoid FI. There is a need to promote effective management strategies for FI to older individuals living in the community. PMID- 16047959 TI - Palliative care nursing education: opportunities for gerontological nurses. AB - Approximately 80% of Americans who die each year are 65 or older. Increasingly, gerontological nurses are asked to deliver high quality end-of-life care. Studies, however, have identified deficiencies in the delivery of care to older adults who are dying-particularly those who die in nursing homes. Enhancing nursing education and training in end-of-life care is one strategy proposed as a remedy for inadequate care for nursing home residents who are dying. This article reviews the current status of end-of-life nursing home care, describes the philosophy and components of quality palliative care, and provides information about opportunities and resources for educating gerontological nurses in end-of life care. PMID- 16047960 TI - Pearl's wisdom. PMID- 16047961 TI - [Care for children born with cleft lip, jaw and/or palate]. AB - The number of children with cleft lip and/or palate needing care in and the possibilities of care delivering to these patients by muitidisciplinary teams are described. The data are compared with data from other European countries. The improvements in quality of care and patients are elucidated by research data. PMID- 16047962 TI - [Dissertations 25 years after the date 5. The dentist and patient education: 25 years later]. AB - In 1979 the thesis 'The dentist and patient education' was published. Based on theory and research, it was the aim of the thesis to provide physicians, dentists, dental students, and dental hygienists theoretical and practical tools to be of assistance in daily patient education. During the last 25 years the insights in patient education and shared decision making were further developed. Two literature studies were published in books. Some research projects were carried out and published in articles. Nowadays, individual based patient education is an essential part of dentists' and dental hygienists' daily work. When compared with the period 1970-1985, at present, multimedia campaigns on oral disease prevention are extremely scarce. Communication skills of dentists appeared to be strongly related to patients' satisfaction of oral care delivery. In the Netherlands, the ethical principle of informed consent has been legislated in 1995 and, as a consequence, activities in the field of patient education are more juridically characterized. PMID- 16047963 TI - [Persistant complaints after endodontic treatment]. AB - A 51-year-old man was referred to an oral and maxillofacial surgeon because of persistent complaints after endodontic treatment of tooth 22. Apical endodontic surgery was performed and periapical tissue was examined histopathologically, showing an osteosarcoma. In retrospect, signs of malignancy were already present on the radiograph taken prior to the endodontic treatment. PMID- 16047964 TI - [Development, physiology, and cell activity of bone]. AB - Bones are of crucial importance for the human body, providing skeletal support, serving as a home for the formation of haematopoietic cells, and reservoiring calcium and phosphate. Long bones develop by endochondral ossification. Flat bones develop by intramembranous ossification. Bone tissue contains hydroxyapatite and various extracellular proteins, producing bone matrix. Two biological mechanisms, determining the strength of bone, are modelling and remodelling. Modelling can change bone shape and size through bone formation by osteoblasts at some sites and through bone destruction by osteoclasts at other sites. Remodelling is bone turnover, also performed by osteoclasts and osteoblasts. The processes of modelling and remodelling are induced by mechanical loads, predominantly muscle loads. Osteoblasts develop from mesenchymal stem cells. Many stimulating factors are known to activate the differentiation. Mature osteoblasts synthesize bone matrix and may further differentiate into osteocytes. Osteocytes maintain structural bone integrity and allow bone to adapt to any mechanical and chemical stimulus. Osteoclasts derive from haematopoietic stem cells. A number of transcription and growth factors have been identified essential for osteoclast differentiation and function. Finally, there is a complex interaction between osteoblasts and osteoclasts. Bone destruction starts by attachment of osteoclasts to the bone surface. Following this, osteoclasts undergo specific morphological changes. The process of bone destruction starts by acid dissolution of hydroxyapatite. After that osteoclasts start to destruct the organic matrix. PMID- 16047965 TI - An overprotected fire service? PMID- 16047966 TI - How to handle the post-inspection process. PMID- 16047967 TI - Mind over matter. PMID- 16047968 TI - 6I sinks in. PMID- 16047969 TI - Plenty to chew on. PMID- 16047970 TI - Boosting survival rates. PMID- 16047971 TI - Noise, under control. PMID- 16047972 TI - Ear muffs: a field guide. PMID- 16047973 TI - Open channels. PMID- 16047974 TI - Serious injury prevention. PMID- 16047975 TI - From compliance to excellence. PMID- 16047976 TI - Standing room only. PMID- 16047977 TI - Taking an upstream approach. Interview by Jerry Laws. PMID- 16047978 TI - Clean sweep. PMID- 16047979 TI - True believers. Interview by Jerry Laws. PMID- 16047980 TI - Guide to hand protection, 2005. PMID- 16047981 TI - More than a cut above. PMID- 16047982 TI - 'Double up' on eye/face protection. PMID- 16047983 TI - Best practices: ergonomic standing surfaces for workers. PMID- 16047984 TI - Incorporating generational diversity. PMID- 16047985 TI - Thoughts on writing a letter to the editor. AB - The "Letters to the Editor" column in any journal is an excellent place to begin writing for publication. Opinions, clarifications, and reflections about articles can be valuable to other readers. Does an article get you thinking? If so, write a letter to the editor. The AORN Journal readers looks forward to hearing from you. PMID- 16047986 TI - Wise use of perioperative antibiotics. AB - NOSOCOMIAL SURGICAL SITE INFECTIONS (SSIs), especially those caused by antibiotic resistant organisms, are a major concern for perioperative nurses. Administering the correct antibiotic at its effective dose within the optimal time frame can help prevent SSIs. THE SIGNIFICANT RELATIONSHIP between microbial resistance and overuse of antibiotics justifies an examination of practices and a move away from widespread antibiotic administration. NEW ANTIBIOTIC GUIDELINES and research support recommendations for antibiotic administration for specific cardiac, colorectal, gynecologic, ophthalmologic, orthopedic, and urinary surgical procedures. Perioperative nurses can help protect their patients from nosocomial SSIs by using this clinical evidence. PMID- 16047987 TI - Systematic review as an evidence-based tool for the operating room. AB - THIS ARTICLE DETAILS advantages of using systematic reviews (ie, meta-analyses) to determine evidence-based best practices. A NUMBER OF SYSTEMATIC REVIEWS relevant to the OR and produced under the auspices of the Cochrane Collaboration are explored with the aim of encouraging perioperative nurses to become familiar with, if not part of, the process of systematic review development. PMID- 16047988 TI - Using an organizational culture analysis to design interventions for change. AB - CULTURAL ASSESSMENTS can be used to make overall generalizations about an organization's culture, identify recommendations for change, create an intervention design to implement change, and develop a communication plan to support implementation. THIS ARTICLE DESCRIBES a cultural assessment of an organization. The perioperative manager and staff members used data, theoretical concepts, and relevant research to identify and implement an organizational change for a medical center's perioperative area. PMID- 16047989 TI - Experiential learning--a theoretical framework for perioperative education. AB - A PERIOPERATIVE NURSING COURSE that uses Kolb's experiential learning theory and learning cycle as a framework was developed to provide nurses new to the OR with a solid knowledge base with which to further their practice. THIS FRAMEWORK can be used to leverage learning style, promote critical thinking, and encourage active learning. PMID- 16047990 TI - High reliability theory and highly reliable organizations. PMID- 16047991 TI - Are nurses suited to new ECP role? PMID- 16047992 TI - It is time for a rethink on rehabilitation. PMID- 16047993 TI - Matrons then and now. PMID- 16047994 TI - Mapping out improved care. Meet the nurse-led dementia care mapping project. PMID- 16047995 TI - The nurse's role in referring patients with suspected cancer. AB - There are anomalies in the UK cancer service, such as varying mortality rates in different parts of the country. New guidance from the National Institute for Health and Clinical Excellence (NICE) seeks to clarify best practice for referring patients with suspected cancer to specialist services. This article looks at the practice implications of this guidance for nurses. PMID- 16047996 TI - Using alcohol handrubs. PMID- 16047997 TI - Alginate dressings. PMID- 16047998 TI - The use of insulin to improve treatment in type 2 diabetes. AB - Diabetes is a global problem of increasing proportions and requires adherence to tight treatment targets to prevent complications. This article focuses on the use of insulin to gain good glycaemia control in type 2 diabetes. PMID- 16047999 TI - A collaborative approach to reducing stress among staff. AB - Stress has been identified as an important issue among residential carers looking after individuals with learning disabilities. This article describes the implementation of a stress-management programme on a residential site for people with learning disabilities, which significantly reduced levels of anxiety and burnout. PMID- 16048000 TI - The evidence on how nurses approach risk assessment. AB - Over the past decade there have been a number of public enquiries that have forced nurses to improve their decision-making skills in risk assessment. This paper discusses the current evidence for how nurses assess risk in everyday practice and concludes that more research is needed to establish protocols for best practice. PMID- 16048001 TI - How to manage a project. PMID- 16048002 TI - Continue to earn as you travel abroad. PMID- 16048003 TI - Do women really want the top jobs? PMID- 16048004 TI - Organic farmers are required to medicate sick animals. PMID- 16048005 TI - Incorrect answer in December 2004 quiz. PMID- 16048006 TI - Cross Canada Disease Report revisited. PMID- 16048007 TI - An ethicist's commentary on trimming of lame dairy cattle. PMID- 16048008 TI - Strain typing of Mycobacterium bovis from a 1978 case of tuberculosis in a wolf (Canis lupis) from Manitoba. PMID- 16048009 TI - Rupture of the peroneus tertius tendon in 27 horses. AB - The purpose of the study was to identify factors influencing the outcome and prognosis of rupture of the tendon of the peroneus tertius muscle in 27 horses. Information on history, physical examination findings, diagnosis, treatment, and final outcome was summarized from medical records. Long-term follow-up information on horses was obtained by telephone survey. A stepwise logistic regression model was used to analyze factors influencing the outcome. Rupture occurred in the midbody of the tendon in 11 horses, at the insertion site in 11 horses, and at the origin in 2 horses. Overall, 18/23 (78.3%) horses returned to their previous level of exercise, 5/23 (21.7%) horses were euthanized due to persistent lameness. If the horse was racing at the time of injury or had an additional structure injured besides the peroneus tertius tendon, it was less likely to return to its intended use. PMID- 16048010 TI - A cross-sectional study of the causes of morbidity and mortality in farmed white tailed deer. AB - Two questionnaires were designed and administered. The first was to a random sample of 340 farmers of white-tailed deer (Odocoileus virginianus) in Canada and the United States. The second was a 10-year retrospective survey of deer submissions to veterinary diagnostic pathology laboratories in Canada and the United States. One-year rates of mortality and common causes of morbidity and mortality for the deer are reported. The primary diagnosis for each record was used to classify diseases into categories, such as parasitic, infectious, toxicological, and neoplastic. Submissions were further classified according to the anatomical location, the pathological change, and the etiology associated with each lesion. Trauma was the most important reported cause of farmed white tailed deer mortality; necrobacillosis was a major cause of morbidity and mortality, especially in fawns. PMID- 16048011 TI - Bacteriological evaluation of commercial canine and feline raw diets. AB - Twenty-five commercial raw diets for dogs and cats were evaluated bacteriologically. Coliforms were present in all diets, ranging from 3.5 x 10(3) to 9.4 x 10(6) CFU/g (mean 8.9 x 10(5); standard deviation 1.9 x 10(6)). Escherichia coli was identified in 15/25 (64%) diets; however, E. coli O157 was not detected. Salmonella spp. were detected in 5/25 (20%) diets; 1 each of beef-, lamb-, quail-, chicken-, and ostrich-based diets. Sporeforming bacteria were identified from 4/25 (16%) samples on direct culture and 25/25 (100%) samples using enrichment culture. Clostridium perfringens was identified in 5/25 (20%) samples. A toxigenic strain of C. difficile was isolated from one turkey-based food. Staphylococcus aureus was isolated from 1/25 (4%) diets. Campylobacter spp. were not isolated from any of the diets. PMID- 16048012 TI - Detection of clusters of Salmonella in animals in Ontario from 1991 to 2001. AB - The temporal patterns of Salmonella serovars isolated from animals in Ontario by 2 major laboratories between 1991 and 2001 were identified. Overall, the number of isolates remained relatively stable and the more frequent isolates were dominant, as in earlier surveys. However, the temporal patterns of specific isolates and the serovars isolated differed depending on the laboratory, species of animal, and reason for obtaining the culture (monitoring versus diagnostic sample). A number of temporal clusters were identified, but their dates of occurrence differed by laboratory. Whereas the laboratories serve an essential role for diagnosis and monitoring, the summary information should be interpreted cautiously. The importance of additional information, such as demographic source, specimen type, and appropriate denominator, when interpreting the data is discussed. PMID- 16048013 TI - Endometrial polyps in 2 African pygmy hedgehogs. AB - Reports of spontaneously occurring endometrial polyps in animals are rare and have only involved a few species. This report is intended to advise veterinarians that older African pygmy hedgehogs may develop endometrial polyps and that these lesions can be a cause of bloody vaginal discharge, sometimes interpreted as hematuria. PMID- 16048014 TI - Sudden death due to thoracic lymphoma in a standardbred racing horse. AB - During training, a 6-year-old standardbred gelding collapsed, exhibiting severe epistaxis and agonal breathing and was euthanized. The horse had a recent history of poor performance, lethargy, and unilateral eyelid swelling with serous ocular discharge. Postmortem and histopathologic examination findings revealed thoracic lymphosarcoma and concurrent exercise-induced pulmonary hemorrhage. PMID- 16048015 TI - Lymphoplasmacytic gingivitis in a cat. AB - A 12-year-old male neutered short haired cat was presented due to difficulty eating and pawing at the face. Examination revealed severe gingivitis and stomatitis throughout the oral cavity. Gingival biopsy provided a diagnosis of lymphoplasmacytic stomatitis. Extraction of all premolars and molars resulted in elimination of all clinical signs. PMID- 16048016 TI - Veterinary diagnostic imaging. Cholestasis as a result of compression of the common bile duct. PMID- 16048017 TI - "Good samaritans": a legislative solution for mandatory reporting of suspected animal abuse. PMID- 16048018 TI - The art of terminal illness and injury: the veterinarian. PMID- 16048019 TI - Coxofemoral luxations--Part 2: Surgical options. PMID- 16048020 TI - Diagnostic dermatology. Zinc responsive dermatosis. PMID- 16048021 TI - Taking the 'LEED'. Environmentally friendly design and construction earns distinction. Boulder Community Foothills Hospital, Colo. PMID- 16048022 TI - Analyze this! Applying Six Sigma concepts to facilities maintenance. PMID- 16048023 TI - Ten lessons learned. Two industry veterans draw major conclusions from high profile health facility projects. PMID- 16048024 TI - Firm foundation. Successful facility expansion is built on robust MEP master planning. PMID- 16048025 TI - 'Surgology' is coming! Designing for the convergence of surgery and interventional radiology. PMID- 16048026 TI - Clean construction. The ES department's role in health facility building projects. PMID- 16048027 TI - [Treatment of bowel obstruction in our own experience]. AB - The intestinal obstruction accounts for the 20% of the acute surgical affections and represents one of the most common indications for the urgent surgery. The clinical scenarios of the intestinal occlusion are rather diversified, with a series of signs not always contemporary present, since they are related to the site (left colon in most cases) and type (neoplastic, by adhesions, or hernia complication) of the obstruction. The authors report their experience about 149 patients clinically observed and treated along a period of 7 years and 3 months. PMID- 16048028 TI - [Orexin, sleep and appetite regulation: a review]. AB - Since 1998, two research groups independently reported the discovery of two novel hypothalamic neuropeptides, called hypocretin-1 and hypocretin-2, respectively many studies have been carried out about their possible functions. One group named these new peptides orexins (A and B) after the Greek word for appetite, since they found that central administration of orexins potently increased food intake in rats. However hypocretins/ orexins are involved in various hypothalamic mechanisms, such as energy homeostasis, neuroendocrine functions, appetite and satiety regulation, vigilance and defence behaviour, sleep and wake regulation. Here is a review of the recent literature, showing some recent discoveries about the link between orexin system, sleep regulation and appetite regulation. PMID- 16048029 TI - [Grapefruit juice: potential drug interaction]. AB - More than a decade has passed since it was unintentionally discovered that grapefruit juice interacts with certain drugs. The coadministration of these drugs with grapefruit juice can markedly elevate drug bioavailability, and can alter pharmacokinetic and pharmacodynamic parameters of the drug. The predominant mechanism for this interaction is the inhibition of cytochrome P-450 3A4 in the small intestine, resulting in a significant reduction of drug presystemic metabolism. An additional mechanism is the inhibition of P-glycoprotein, a transporter that carries drug from the enterocyte back to the gut lumen, resulting in a further increase in the fraction of drug absorbed. Some calcium channel antagonists, benzodiazepines, HMG-CoA reductase inhibitors and cyclosporine are the most affected drugs. A single exposure to one glass of the grapefruit juice can usually produce the maximal magnitude of the interaction. The data available so far, concerning this interaction and its clinical implications, are reviewed in this article. It is likely that more information regarding this interaction will accumulate in the future, and awareness of such is necessary for achieving optimal drug therapy. PMID- 16048030 TI - [Gilles de la Tourette's syndrome: a systematic review]. AB - Gilles de la Tourette's syndrome is more frequent than once believed. This syndrome is a chronic disorder whose long term outcome is generally favourable, characterized by a fluctuating course. The etiopathogenesis of Gilles de la Tourette's syndrome has not been ascertained, although the frontal-subcortical neural pathways seem to be involved. This extrapyramidal syndrome is frequently associated with attention-deficit/hyperactivity disorder, obsessive-compulsive disorder, and behaviour problems. A correct diagnosis is the first step for a proper management of this disorder, which makes use of behavioural and pharmacological interventions. PMID- 16048031 TI - [Use of levosimendan after cardiac arrest by ventricular fibrillation: a case report]. AB - We describe the case of a patient admitted in intensive care, after cardiac arrest by ventricular fibrillation treated by electrical defibrillation, that showed a serious deficit of the cardiac index and increase of the systemic vascular resistances. The patient was treated by levosimendan (Simdax): a starter dose of 12 microg/Kg in ten minutes and then a continuous perfusion of 0.1 microg/Kg/min. for 24 hours. He had a continuous amelioration of the clinical conditions and of the hemodynamics parameters. In the fifth day the patient was transferred to the Cardiology department and after 20 days he was discharged from hospital. PMID- 16048032 TI - [Diet, nutrition and rheumatoid arthritis]. AB - Rheumatoid Arthritis (RA) is a chronic inflammatory disease resulting in diarthrodial joints inflammation (particularly joints of hands, wrists, feet, knees, cubitus, ankles, shoulder, etc.) that is manifested by swelling and functional impairment. The associated complications, osteoporosis and cardiovascular disease, make RA important in public health terms. During the active phase of disease, elevated plasma concentrations of inflammatory cytokines, such as interleukin-6 (IL-6), interleukin-1beta (IL-1beta), tumour necrosis factor-alpha (TNF-alpha) and acute-phase proteins, lead to reduction of fat free body mass (FFM) with a loss mean of 15% of cell body mass (CM) and consequent reduction of muscle strength. The pharmacological therapy (non steroidal anti inflammatory drugs (NSAIDs), slow acting antirheumatic drugs and corticosteroids), have the potential to cause side-effects, such as gastrointestinal bleeding, bone loss beyond to increase the requirement of some nutrients and reduce their absorption. The diet may play role in the management of RA, particularly in alleviating the symptoms of the disease, combating the side-effects of therapy and reducing the risk of complications. The increase of the caloric and proteic intake is not sufficient to offset a increased metabolic rhythm and important proteic catabolism but a diet balanced may warrant an adequate intake of nutrients. The carbohydrates of the diet provide 55-60% of the caloric intake, the diet is normo-proteinic or hyper-proteinic in the active phase of disease, and lipids represent 25-30% of the caloric intake (saturated, monounsaturated, polyunsaturated fatty acids in the ratio 1:1:1). omega-3 fatty acids supplementation, in combination with reduction of fatty acids omega-6 and adequate intake of monounsaturated fatty acids induce improvement in symptoms and sometimes a reduction in NSAIDs usage. Proper antioxidant nutrients (Vitamin A, Vitamin C, selenium) may provide an important defence against the increased oxidant stress and a supplementation of folate and vitamin B12, in patients treated with methotrexate (MTX), reduce the incidence of side effects and offset the elevation in plasma homocysteine frequent in these patients. Calcium and vitamin D, in patients treated with corticosteroids, reduce the bone loss, while a supplementation with iron may prevent anaemia. Finally, elimination diets may be feasible therapy only in patients with positive skin prick test. PMID- 16048033 TI - [Essential thrombocythemia]. AB - New diagnostic and therapeutic aspects of the essential thrombocythemia are summarized. A series of 14 patients with essential thrombocythemia is reported. PMID- 16048034 TI - Looking for parasitic infection and disease: the Plasmodium falciparum malaria model. PMID- 16048035 TI - [Relationship between oxidant stress and milk productivity in dairy cows]. AB - An imbalance between formation and detoxification of oxygen radicals leads to oxidant stress that may increase in more intense oxidative metabolism caused by a high intake of metabolizable energy to provide metabolic intermediates for the milk synthesis and secretion. This hypothesis was tested using dairy cows and the concentration of hydroperoxides in lipids (LHP) extracted from circulative lipoprotein particles of low and very low density (LDL and VLDL/chylomicrons) as oxidant stress indicator. The particles were prepared by ultracentrifugation of serum obtained by coccygeal bleeding (13 cows, 1. parity, n=8 and 2. parity, n=5, lactation stage, 53 +/- 1.4 days post partum) and purified by precipitation. Concentrations of LHP-LDL/mg Lipoprotein correlated significantly with daily milk yield (r = 0.73, P = 0.004) or daily milk energy output (r = 0.77, P = 0.003) in contrast to LHP of VLDL/chylomicron particles. Thus, some evidence was obtained for an almost linear, positive relationship between milk productivity and oxidant stress occurring in LDL. PMID- 16048036 TI - [Evaluation of foot pad health of laying hens in small group housing systems and furnished cages]. AB - Foot pad health was examined in a total of 648 Lohmann Selected Leghorn (LSL) in two trials and in 216 Lohmann Brown (LB) laying hens in one trial kept in small group housing systems and furnished cages. The effects of housing system, strain, month of laying period, rearing method, group size and body weight were tested for significance by using analysis of variance. The assessment of foot pads was performed each three months during the laying period. The alterations of foot pads were classified in severity on a scale from 1 (no hyperkeratosis, no epithelial lesions, no swelling of foot pads) to 5 (severe hyperkeratosis and/or deep and large epithelial lesions and/or moderate or high-grade swelling of foot pads). In addition to the macroscopic findings, foot pads were histologically examined. Pathological alterations of foot pads were found in 86.1% of the laying hens, whereas 57.4% of the examined animals showed mild hyperkeratosis (degree 2). Modest hyperkeratosis and/or superficial lesions of the epithelium (degree 3) were detected in 21.0% of the laying hens examined. Severe hyperkeratosis and/or deep epithelial lesions and/or mild swelling of the foot pads (degree 4) as well as very severe hyperkeratosis and/or deep and large epithelial lesions and/or moderate or high-grade swelling of foot pads (degree 5) occurred with frequencies of 5.9% and 1.9% respectively. LB laying hens' food pads were more frequently affected by hyperkeratosis, whereas epithelial lesions were more frequently found in LSL laying hens. The prevalence of pathological alterations in foot pads was significantly influenced by housing system and month of laying period. The results of this study showed that laying hens kept in small group housing systems and furnished cages develop proliferative hyperkeratosis in foot pads due to increased mechanical compression load, caused by the use of perches and staying on wire floor. PMID- 16048037 TI - [Measurement of temperature generation in the corium of bovine claws during claw trimming with an angle grinder using different grinding and cutting disks]. AB - On 80 bovine hindclaws from slaughtered cows a correct functional claw trimming (sole horn thickness 5 mm; group 1) and on further 80 hindclaws a standardised, incorrect claw trimming (sole horn thickness 2 mm; group 2) was performed using 4 different disks (one cutting, three grinding disks). Two thermocouples were inserted into the solar corium and the heat production on the corium was measured during claw trimming. During correct functional claw trimming with each of the 4 disks a temperature increase in the corium with a median of < or = 0.3 degrees C was calculated. During incorrect claw trimming (2 mm sole horn thickness) a temperature increase with a median of < or = 0.3 degrees C for the first three disks was determined. During claw trimming using the abrasive semiflexible plastic disk a median of 1.3 degrees C temperature increase was calculated. Comparison of the temperatures measured in the claws from group 1 and 2 showed a statistically significant difference between all four disks. Comparison of the temperature increase within each group (group 1 and 2) revealed a statistically significant difference between the cutting disk with steel blades and the abrasive, semiflexible plastic disk in both groups. The highest temperatures measured were 41.5 degrees C in group 1 using the disk with 70% amount of fitted hard metal granulate, and 42.2 degrees C in group 2 (temperature increase of 10.6 degrees C) using the abrasive semiflexible plastic disk. Under the experimental conditions of this study, only a very slight thermogenesis was found when a functional and correct claw trimming using the 4 tested disks was performed. This slight thermogenesis should not be able to cause thermic injury of the corium. By trimming the sole horn incorrectly (2 mm) the measured heat generation in the corium was minor, without danger of thermic insult, excepted for using the disk with a 70% amount of fitted hard metal granulate and the abrasive semiflexible plastic disk. Even, it is unlikely that a short term temperature increase of 10.6 degrees C can cause thermic insults of the solar corium. PMID- 16048038 TI - [Laboratory diagnosis of avian influenza by reverse transcription (RT)-PCR]. AB - RT-PCR assays which amplify conserved regions of the influenza A virus gene are useful tools for the rapid and specific detection of infections of poultry with avian influenza virus (AIV) and for the investigation of large numbers of samples, e.g. within the framework of surveillance programs. Here, we present findings on the efficiency and on the limits of an RT-PCR assay which amplifies a part of the matrix protein gene. Sensitivity and specificity of the method were increased by the additional use of nested PCR. Parameters which may have an essential influence on the detection limit are outlined and discussed. A major focus of the study is the detection of AIV RNA from organ samples and swabs. PMID- 16048039 TI - Clostridium novyi caused outdoor sow mortality in croatia. AB - In a Croatian outdoor pig breeding unit 32 sows (died between days 2 and 14 post partum) were subjected to gross pathological and further laboratory investigations. Necropsy findings revealed tympany and purple discoloration of the skin, the surface of the livers was dark and had honeycomb appearance with gas bubble infiltrations, congested lungs, hemorrhages, serosanguinous exsudates in body cavities and the stomachs were full. Gram-stains of smears revealed large numbers of Gram-positive rods. Anaerobic cultures yielded high numbers of Clostridium (C.) novyi and fluorescent antibody test (FAT) confirmed this diagnosis. Enzyme immunoassay and toxin testing by neutralisation in Chinese hamster ovary cells revealed toxin B. Based upon the clinical symptoms, gross pathological signs, bacteriological findings and toxin testing we concluded that C. novyi caused sow mortality. Suboptimal outdoor environment and high outdoor infectious pressure might have contributed the C. novyi caused losses in this unit. PMID- 16048040 TI - Molecular detection and characterization of Ehrlichia canis from dogs in Turkey. AB - Seroprevalence of Ehrlichia canis antibodies among dogs in Turkey were previously reported, however, the ehrlichial organism has never been characterized in this region. The current study examined dogs from Ankara with febrile illness for E. canis infection with E. canis-specific PCR. Three of the 12 blood specimens from dogs showing clinical signs compatible with canine ehrlichiosis were found to be positive by PCR using E. canis-specific primers. E. canis detected in one of the blood specimens was designated as Kutahya strain. The representative E. canis strain was characterized by 16S rRNA gene sequencing and Western blot analysis of the plasma sample from the dog infected with E. canis. The 16S rRNA sequence (1,388 bp) of the E. canis Kutahya was identical to that of Ehrlichia ovina from a sheep in Turkey and Venezuelan Dog Ehrlichia (VDE) and was closely related (99.9%) to that of type strain of E. canis, Oklahoma. The plasma of the dog infected with E. canis Kutahya was analyzed by Western blotting using the purified E. canis Oklahoma strain as antigen. The reactive antibody profiles of the dog infected with E. canis Kutahya was found to be similar to those of dogs infected with E. canis Oklahoma and VDE, suggesting the antigenic similarities among these strains. The findings in this study would help for a better understanding of epidemiology of canine ehrlichiosis. This is the first report of molecular detection and characterization of an ehrlichial agent in Turkey. PMID- 16048041 TI - Effect of steroidal and non-steroidal anti-inflammatory drugs on inflammatory markers in calves with experimentally-induced bronchopneumonia. AB - The influence of treatment with steroidal (SAIDs) and non-steroidal (NSAIDs) anti inflammatory drugs on inflammatory markers in thirty, 6-8 week old calves with induced bronchopneumonia was investigated. Animals received a single intravenous treatment with meloxicam (0.5 mg/kg body weight), flumethasone (0.05 mg/kg body weight) or sterile 0.9% NaCl (10 ml per animal). Body temperature, respiratory and heart rate, concentration of prostaglandins PGE2, PGF2alpha, thromboxane (TXB2), leukotriene (LTB4) and malonyldialdehyd (MDA) and proinflammatory cytokines i.e. tumor necrosis factor (TNFalpha) and interferon (INFalpha) were recorded in serum, bronchoalveolar lavage (BAL) and blood platelets (BP). A significant reduction of main inflammatory mediators PGE2, PGF2alpha,TXB2 and MDA after meloxicam treatment in calves with induced bronchopneumonia indicates a beneficial effect on the inflammatory processes. Contrary to effects observed by flumethasone, meloxicam induced an increase of LTB4 and INFalpha indicating that it is not immunosuppressive. PMID- 16048042 TI - [The effect of response behaviour on veterinary epidemiological studies--an analysis based on a cross-sectional study of livestock management of cattle farms in Lower Saxony]. AB - Cross-sectional studies are used as an epidemiological instrument in several veterinary investigations. A common means of collecting information is the questionnaire, which is send to diagnostic units like farmers. A common fact is, that not all designated individuals take part. So it is to be clarified, if and in which concern this fact influences the results of an epidemiological study. Considering the example of a cross-sectional study in cattle farms this paper shows, that consistency of Responders and Non-Responders differs significantly. Technical measures to estimate this effect and consequences for the results are shown. Small and medium farms showed a different response rate than big farms before as well as after a mailed reminding letter. Considering the complete return period there was no significantly different response rate between farms of different sizes. Furthermore there are noticeable differences between Responders and Non-Responders concerning parameters like housing or use of commercial feedstuff. Therefore it is implicitly required to send a reminding letter and pursue Non-Responders in researches. Otherwise a biased sample collective is generated. PMID- 16048043 TI - [Mycobacterium avium subsp. paratuberculosis in wild animal species and cattle in Styria/Austria]. AB - Infections with Mycobacterium ovium ssp. paratuberculosis (M. paratuberculosis) are increasingly recognised worldwide. In addition to an increased prevalence of paratuberculosis in Austrian cattle herds, recent years have also shown a rise in infections with M. paratuberculosis in wild red and roe deer, chamois and mouflon. During the period from June 2002 to September 2004, mesenteric lymph nodes were taken from a total of 483 wild animals hunted or found dead and from 338 deceased cattle. Samples were analysed using PCR and cultivation methods. In the case of pathomorphological changes or anamnestic indications, investigations also included an analysis of organ samples (e.g. liver, lung) or foetuses. The tests revealed that 129 wild animal samples (red deer, roe deer, chamois, mouflon, fallow deer, ibex, foxes, mountain hare, yellow-necked field mouse, and capercaillie) contained M. paratuberculosis. The major symptoms in the wild aninodes. Evidence of diarrhoea was only observed in about 15% of the positive cases. The study for the first time provided evidence of intrauterine transmission of M. paratuberculosis in red deer (3 cases) and chamois (1 case) and succeeded in the isolation of the pathogen from the liver, lung and subcutaneous granulomas of wild animals. Of the total of 338 mesenteric lymphnodes of cattle from 303 herds, 80 samples from 77 herds tested positive for paratuberculosis. Twenty-two wild animal and 3 cattle isolates have so far been molecularly typed using IS900-RFLP and RAPD analyses in order to prove epidemiological relationships between occurrences in cattle and wild animals. The increase of paratuberculosis in wild animal species is assumed to have been caused by the purchase of animals, a strong increase in suckler cow farming (cow calf herds) with a concentration of pathogens in the environment and by inadequate feed hygiene for wild animals. PMID- 16048044 TI - [Occurrence of Salmonella spp. and shigatoxin-producing escherichia coli (STEC) in horse faeces and horse meat products]. AB - In order to assess the relevance of horses as a possible reservoir of Salmonella and Shigatoxin-producing Escherichia coli (STEC), 400 samples of horse faeces and 100 samples of horse meat products were examined by PCR-screening methods. Salmonella enterica was not found in any of the samples. One faeces-sample and one horse meat product were proved to be STEC positive. The STEC-strain from faecal origin belonged to the serotype 0113:H21 and had the stx 2c gene and the enterohemolysin gene. The STEC-strain isolated from a horse meat product had the serotype O87:H16 and the stx 2d gene. The results indicate a very low risk for human to get a Salmonella- or EHEC- infection from horses in Germany. PMID- 16048045 TI - [Effect of beta-carotin serum concentration on the reproductive performance in dairy cows]. AB - The aim of the following investigation was to study the effect of beta-carotin serum concentrations on reproductive performance in dairy cows. Blood samples were collected from 201 cows 3 to 4 weeks antepartum, 1 to 2 weeks postpartum and at the time of insemination. Concentrations of beta-carotin in blood serum were determined via HPLC. The incidence of retained placental fetal membranes, endometritis, ovarian cysts as well as the beginning of the oestrous cycle and various measures of fertility such as first service conception rate, pregnancy index, interval from calving to first insemination, interval from calving to conception, interval from first insemination to conception and expected calving interval were recorded and analysed. The beta-carotin serum concentrations were not related to the incidence of retained placental fetal membranes, endometritis, ovarian cysts or the onset of cyclicity post partum. Cows with lower beta-carotin concentrations partially seemed to have a reduced reproductive performance than cows with higher beta-carotin concentrations. Overall, our results suggest only a minor relationship between the beta-carotin serum concentration and fertility in dairy cows. PMID- 16048046 TI - The hardness of horn in different segments of the bovine claw. AB - Hardness of bovine hoof horn was tested as ball indentation hardness and as shore D hardness post mortem in different segments of the hoof wall, in the sole and the hard bulb of sound claws of 10 Austrian Holstein Friesian cows. Both methods of hardness determination showed corresponding results, with shore D hardness between 52.2 and 63.9 hardness units (hu) and ball indentation hardness between 11.2 N/mm2 and 24.3 N/mm2. Bovine hoof horn becomes significantly softer from the coronary band towards the weight bearing border (vertical decrease) and from the dorsal wall towards the heel (horizontal decrease). Decreasing hardness was associated with decreasing dry matter content. Measurements of the claw capsule showed thickness of the hoof wall increasing from the coronary border towards the sole. In dorsopalmar/-plantar direction, bovine hoof wall at the weight bearing border decreases towards the heel. PMID- 16048047 TI - Investigation of diagnostic importance of platelet closure times measured by Platelet Function Analyzer--PFA 100 in dogs with endotoxemia. AB - This study was performed to evaluate the diagnostic importance of the platelet closure times measured by the Platelet Function Analyzer (PFA-100) in dogs with endotoxemia. E. coli endotoxin was given intravenously once, at the dose of 0.02 mg/kg or 1 mg/kg in groups I (n=9) and II (n=8), respectively. Normal saline (0.1 ml/kg) was injected in group III (n=8). The dogs were monitored for 48 h, and venous blood samples were collected prior to (baseline) and at intervals of 0.5, 1, 2, 4, 6, 8, 12, 24 and 48 h subsequent to the treatments. The white blood cell (WBC), platelet counts, and hematocrit (Hct) values were recorded. Platelet closure times were determined, using collagen/epinephrine (CEPI) and collagen/adenosine diphosphate (CADP) cartridges. Within 0.5 h after the endotoxin application baseline WBC and platelet counts (mean +/-SD) decreased significantly (p<0.001) to 2000 +/- 500 and 1850 +/- 200 cells/microl or 69.000 +/- 12.500 and 27.000 +/- 6.400 cells/microl in groups I and II, respectively. Platelet counts remained low during the first 1-48 h, but the WBC count was high at the 8th-48th h, in groups I and II, compared with baselines (p<0.001). After the application of the endotoxin, Hct values increased from baseline values of 37 +/- 3 or 39 +/- 2% to 48 +/- 2 or 51 +/- 3%, within 1 h (p<0.001), in groups I and II, respectively. Hct values in group II were notably higher (p<0.001) than those of group I, during the 2nd-48th h. Hematological parameters and closure times did not differ significantly throughout the study in group III. Baseline closure time ranged from 79 +/- 5 seconds (s) to 86 +/- 5 s for CADP and 144 +/- 13 s to 159 +/- 14 s for CEPI in all dogs (n=25). At 0.5 h after the endotoxin, the closure times of CADP as well as CEPI declined to 62 +/- 6 s and 76 +/- 8 s in group I (p<0.001) and 57 +/- 5 s and 75 +/- 6 s in group II (p<0.001). Afterwards, closure time prolonged to the levels of 280 +/- 8 s (CADP) and 294 +/ 5 s (CEPI) by 48 h (p<0.001) in group II, but returned to the baseline limit in group I. In conclusion, our results show that the shortened closure times may serve as a very early diagnostic sign of endotoxemia, prolonged closure times however may be used as an index for the severity of endotoxemia. PMID- 16048048 TI - Sarcomas of the soft tissues. Thirteen years of experience at the Rizzoli Orthopaedic Institute. PMID- 16048049 TI - Neoadjuvant chemotherapy for the treatment of osteosarcoma of the extremities: a comparison of results obtained in single-institution and multicenter trials. AB - Osteosarcoma is a rare tumor. In multicentric trials on the neoadjuvant treatment of this neoplasm many of the single institutions involved have the opportunity to treat a small number of cases (the average for all of the studies was less than 1 case per year). In order to verify whether this can change the percentage of amputations avoided and prognosis, a review of the current literature was carried out, concerning neoadjuvant treatment of osteosarcoma of the extremities. The results obtained in 9 multicentric trials and in 12 mono-institutional trials were evaluated. It was observed that in mono-institutional studies the percentage of patients treated by conservative surgery instead of aggressive surgery (73% vs 55%) and the disease-free survival 5 years after surgery (73% vs 55%; p < 0.0001) are significantly higher as compared to what was observed for patients treated in multicentric trials. Based on these differences, the authors conclude that it would be opportune to direct patients with osteosarcoma of the extremities to the best centers that each year treat large numbers of patients with musculoskeletal tumors. PMID- 16048050 TI - Vertebral hemi-resection for bone tumor with wide invasion of the vertebral canal: modified surgical method. AB - The authors describe a variation in the method of vertebral hemi-resection used for the treatment of neoplasms that present a wide invasion of the vertebral canal. This is followed by a review of the literature on the subject. PMID- 16048051 TI - Recombinant human erythropoietin in pediatric patients: efficacy in facilitating autologous blood donation in spinal deformity surgery. AB - Preoperative autologous blood donation (PABD) is a widely used practice in orthopaedic elective surgery, but many pediatric patients are unable to complete the program of pre-deposit. Twenty-three consecutive patients undergoing spinal surgery for scoliosis received 6 administrations of 10000U of recombinant human erythropoietin (rHuEpo). Preop hemoglobin (Hb) levels and the numbers of collected and of autologous and allogeneic blood transfused units were determined. These results were compared with a previously-operated group of 28 patients, who differed only by the absence of concomitant erythropoietin therapy. Significant higher numbers of collected blood units and Hb levels were measured, and all of the patients completed the PABD program. A significantly lower requirement for allogeneic blood in the <> group was observed: 1 vs 9 patients (4.3%-32.1%; p < 0.001). The study documents the efficacy of rHuEpo in facilitating the completion of a PABD program and in reducing exposure to allogeneic blood in pediatric patients undergoing corrective spinal surgery. PMID- 16048052 TI - Giant cell tumor of the upper cervical spine: transmandibular-translingual access. Clinical case. AB - The authors describe the clinical case of a patient aged 18 years affected with giant cell tumor (GCT) at C3 who came to the surgical unit of Orthopaedics and Traumatology at the Ospedale Maggiore in Bologna after being treated by surgery elsewhere. Particular attention is paid to surgical access by means of median transmandibuloglossotomy used in order to obtain a sufficiently wide surgical field that can adequately expose the vertebral segment affected by neoplastic disease. In particular, possible complications that may be observed postsurgery can be compared to other surgical approaches to the upper cervical spine and above all that there are no permanent clinical sequelae. PMID- 16048053 TI - The surgical treatment of malleolar fractures: long-term results. AB - The retrospective study concerns use of the AO method for the treatment of 75 ankle fractures between 1990 and 1995. After a mean follow-up of 5 years and 7 months the Cedell criteria were used for a subjective evaluation of results. Good results were obtained in 53 out of 75 patients submitted to surgery (70%), with a significant difference between uni-malleolar fractures and bi- and tri-malleolar fractures. Dorsal flexion measured an average of 24 degrees (80% of the healthy contralateral side), plantar flexion 36 degrees (90% of the healthy side). The presence of a fracture-dislocation considerably worsens prognosis because of the cartilaginous damage produced. The risk of poor results increases considerably when reduction of fracture is insufficient or absent. Post-traumatic arthrosis constitutes a common complication (25%) and it particularly affects females aged between 45 and 65 years, as a result of the presence of post-menopausal osteoporosis. Accurate reduction, fracture stabilization, sex and age constitute essential elements for satisfactory final results. PMID- 16048054 TI - The echo-guided treatment of calcific tendinitis of the shoulder. AB - The authors report the results of percutaneous mini-invasive treatment of chronic calcific tendinitis of the rotator cuffs. A total of 39 patients have been treated by echo-guided injection under local anaesthesia since June 2000 with a follow-up of about 2 years. Considerable reduction in symptoms was obtained in 34 patients within a few days of treatment; improvement was moderate in 5 cases, there were no complications in any of the cases. Complete regression of calcification was observed in 21 patients, there was a more than 60% reduction in calcific deposits in 11 patients, there was little reduction in 5, and the calcification remained unchanged in 2. The method, based on our experience, proved to be simple to execute, low-cost and easily repeatable, offering good results from a symptomatological point of view as well. It is the purpose of this study to determine and maximize mini-invasive treatment that will allow for the elimination or reduction of calcifications by means of percutaneous and echo guided access. PMID- 16048055 TI - A case of Freiberg's disease in an adult patient. AB - Freiberg's disease is an osteochondrosis of the IInd metatarsal head that prevalently develops during the second decade of life and that is the cause of important painful symptoms that resist conservative treatment. The disease is quite rare and must be treated surgically during its early phase in order to prevent progression that may result in permanent changes in the metatarsal head. It is the purpose of this study to describe the clinical case of a patient aged 30 years affected with Freiberg's disease, diagnosed at the age of 15 years, and never submitted to either conservative treatment or surgery. PMID- 16048056 TI - Non-Hodgkin's lymphoma in the gluteal region: a case report. AB - The infiltration of the skeletal muscles on the part of lymphomatous cells is rarely reported and only histological examination can reveal it. Based on a review of the literature, it is the purpose of this study to describe a rare case of non-Hodgkin's lymphoma with muscular invasion, characterized by voluminous swelling in the left gluteal region, in a female aged 60 years who was completely asymptomatic. Biopsy and immunohistochemistry alone allowed us to diagnose non Hodgkin's lymphoma. It is the purpose of this study to document the existence of NHL with musculoskeletal localization, emphasizing the morphological features and the site, as well as hypothesizing that wide-margin surgery can constitute a valid therapeutic aid for such extralymphonodal forms, that are particularly large and with a single localization. PMID- 16048057 TI - Gorham's disease: a rare case of multicentric localization. AB - Gorham's disease is described as a rare form of massive non-familiar idiopathic osteolysis with a self-limiting nature, where the simil-angiomatous tissue destroys the bone, that is substituted by fibrous tissue. One case of Gorham's disease is presented, that is interesting because of the sites in which the pathology is localized and because of its multicentricity. The authors reached a diagnosis by carefully studying clinical and laboratory evidence as well as imaging and anatomo-histopathological findings, confirmed by the data reported in the literature. PMID- 16048058 TI - Aneurysmal bone cyst in children involving infrequent locations. Report on two cases. AB - Aneurysmal bone cyst is a benign but locally destructive lesion of bone. We present two cases of ABC in uncommon locations in two children. The first case concerns a 14 year old boy who had a mass located in the internal side of the talus. An ABC type V of Campanacci's classification is diagnosed. Complete curettage was carried out with a high-speed burr. We implanted an Orthofix external fixation with a mobil bottom bolster and distal pins located in the calcaneum. Case 2 concerns an ABC in the clavicle of a 9 years old boy. A selective arterial embolization (SAE) with polyvinyl alcohol was carried out. Curettage of the lesion and filling with a cortico-spongy graft were implemented. Two years later, both of the patients are asymptomatic. We believe that external fixation with a bolster is an efficient and valid alternative to traditional immobilization with a cast. The use of SAE also prevented profuse bleeding of the lesion during surgical procedure. PMID- 16048059 TI - [Basics for service in occupational medicine: strategy, structure, activities, resources]. PMID- 16048060 TI - [Hygienic evaluation of work conditions and morbidity parameters among workers engaged into oil and gas extraction in Kyrgyz Republic]. AB - The article covers hygienic evaluation of work conditions and morbidity parameters among workers engaged into oil and gas extraction in Kyrgyz Republic. Work conditions are characterized by physical, chemical and psychophysiologic occupational hazards that, if long-lasting, could promote formation of occupational diseases. Results of the study helped to specify prophylactic measures. PMID- 16048061 TI - [Occupational bronchitis caused by aerosol fibers of chrysotile asbestos]. AB - The article represents clinical features, course types and outcomings of occupational bronchitis caused by fibers of chysotile asbestos aerosol. Authors cover clinical, functional, X-ray and hygienic aspects of occupational bronchitis due to contact with dust containing chrysotile and varying in contents. PMID- 16048062 TI - [Conditions of development and course of main clinical pain syndromes in occupational vertebrogenic diseases of lumbosacral level]. AB - The authors studied clinical features concerning formation and course of main clinical syndromes of skeletal and muscular disorders associated with predominant algias. Major risk factors for occupational vertebrogenic diseases on lumbo sacral level are identified. PMID- 16048063 TI - [Forecasting heat and functional state of human exposed to cooling in water medium]. AB - Based on mathematic and statistic analysis of results obtained in studies of human heat exchange with cooling water medium, the authors represented canonical correlational patterns to determine integral parameter of cooling conditions (IPCC) referred to naked human and with various clothes on, both with and without additional heat releasing sources. Mathematic and statistic analysis helped to present correlational patterns for predicting levels of changes in human functional state according to IPCC comprising complex of factors that determine heat exchange in water medium, including safe time for stay in it. PMID- 16048064 TI - [Magnetocardiography as a new diagnostic method for cardiologic diseases in occupational medicine]. AB - Magnetocardiography as a new diagnostic method for cardiologic diseases in occupational medicine. The article covers possibilities of magnetocardiography as a promising diagnostic method for cardiologic diseases in occupational medicine. The studies used 7-channels magnetocardiographic complex MKG-6073-M. The authors applied standard exercise tests in 6 apparently healthy examinees. PMID- 16048065 TI - [Criteria to reasonably choose specific type of individual protective means for respiratory organs in occupational environment]. AB - Use of individual protective means for respiratory organs could be effective in condition of scientifically justified choice of specific design for those means, with consideration of their purpose, protective and exploitative properties, quantitative and qualitative composition of chemicals in ambient air, characteristics and heaviness of occupational tasks. PMID- 16048066 TI - [Occupational Medicine Department in St Petersburg Medical Academy for Postgraduate Education reports on results of practical training of research educational specialists for the nearest foreign countries]. PMID- 16048067 TI - Australian consequentialism: an Australian critique. PMID- 16048068 TI - Peter Singer's argument for utilitarianism. AB - The paper begins by situating Singer within the British meta-ethical tradition. It sets out the main steps in his argument for utilitarianism as the 'default setting' of ethical thought. It argues that Singer's argument depends on a hierarchy of reasons, such that the ethical viewpoint is understood to be an adaptation--an extension--of a fundamental self-interest. It concludes that the argument fails because it is impossible to get from this starting-point in self interest to his conception of the ethical point of view. The fundamental problem is its mixing the immiscible: the Humean subordination of reason to interest with the Kantian conception of reason as universal and authoritative. PMID- 16048069 TI - The prodigal and his brother: impartiality and the equal consideration of interests. AB - At the heart of Peter Singer's utilitarianism is the impartial weighing of the interests of those affected by our actions. Singer calls this the Principle of Equal Consideration of Interests. This paper argues that Singer's Principle does not accord with our moral intuitions and the logic of our moral thinking. It discusses the Principle in the context of the parable of the Prodigal Son and his Brother--a parable that raises the issue of impartiality in a particularly challenging way. What the parable shows is, first, that our moral thinking often turns on judgements of fairness that are prior to any impartial weighing of interests; and, second, that impartial fairness itself is sometimes transcended by compassionate love. Both of these points have important implications for bioethics. PMID- 16048070 TI - Life and death matters: losing a sense of the value of human beings. AB - The essay combines a specific and a more general theme. In attacking 'the doctrine of the sanctity of human life' Singer takes himself thereby to be opposing the conviction that human life has special value. I argue that this conviction goes deep in our lives in many ways that do not depend on what Singer identifies as central to that 'doctrine', and that his attack therefore misses its main target. I argue more generally that Singer's own moral philosophy affords only an impoverished and distorted sense of the value of human life and human beings. In purporting to dig below the supposedly illusion-ridden surface of our thinking about value, Singer in fact often leads us away from the robust terrain of our lived experience into rhetorical, and sometimes brutal, fantasy. PMID- 16048071 TI - Consequentialism, complacency, and slippery slope arguments. AB - The standard problem with many slippery slope arguments is that they fail to provide us with the necessary evidence to warrant our believing that the significantly morally worse circumstances they predict will in fact come about. As such these arguments have widely been criticised as 'scare-mongering'. Consequentialists have traditionally been at the forefront of such criticisms, demanding that we get serious about guiding our prescriptions for right action by a comprehensive appreciation of the empirical facts. This is not surprising, since consequentialism has traditionally been committed to the idea that right action be driven by empirical realities, and this hard-headed approach has been an especially notable feature of Australian consequentialism. But this apparent empirical hard-headedness is very selective. While consequentialists have understood their moral outlook and commitments as guided by a partnership with empirical science - most explicitly in their replies to the arguments of their detractors - some consequentialists have been remarkably complacent about providing empirical support for their own prescriptions. Our key example here is the consequentialist claim that our current practises of partiality in fact maximise the good, impartially conceived. This claim has invariably been made without compelling support for the large empirical claims upon which it rests, and so, like the speculative empirical hand-waving of weak slippery slope arguments, it seems similarly to be undermined. While these arguments have presented us with 'wishful thinking' rather than 'scare-mongering', we argue in this paper that their complacency in meeting the relevant empirical justificatory burden remains much the same. PMID- 16048072 TI - Why do they hate us, thick and thin? AB - Immediately after the September 11, 2001 terrorist strikes on the World Trade Center Towers, the Pentagon, and a plane over Pennsylvania, many in the West, but particularly the United States of America, felt urgently the pain of the question 'Why do they hate us?' in relation both to those who directly perpetrated those dreadful events and to those who sympathised with their perpetrators. In this paper, I will offer an account of some of the conceptual issues at stake in addressing seriously such a question as an opportunity for self-examination. I will argue that questions of this kind are the very warp and weft of international relations at their most serious levels and that the often glib demonisation of 'them' who hate us serves us badly in coming to terms with the problems the West faces in its relations with the Muslim world. I use this most pressing of international relations issues as a test of the prospects of Utilitarianism as a viable 'ethics of international relations' arguing that its notions of the good and its resources for criticism of desires and preferences are a cause for scandal in our inter-cultural negotiations with other traditions of civilization, particularly the Muslim civilization. In many ways, rather than being part of the solution, Utilitarianism is part of the problem. PMID- 16048073 TI - Pettit on consequentialism and universalizability. AB - Philip Pettit has argued that universalizability entails consequentialism. I criticise the argument for relying on a question-begging reading of the impartiality of universalization. A revised form of the argument can be constructed by relying on preference-satisfaction rationality, rather than on impartiality. But this revised argument succumbs to an ambiguity in the notion of a preference (or desire). I compare the revised argument to an earlier argument of Pettit's for consequentialism that appealed to the theoretical virtue of simplicity, and I raise questions about the force of appeal to notions like simplicity and rationality in moral argument. PMID- 16048074 TI - [Orthopedics in transition. The evolution of the discipline and professional status among peers and in the Kaiser empire (1815-1914)]. PMID- 16048075 TI - Honor among beasts. Think altruism, empathy and a sense of fair play are traits only humans possess? Think again. PMID- 16048076 TI - Water works. PMID- 16048077 TI - Giving all you can. PMID- 16048078 TI - Board's eye view. PMID- 16048079 TI - The future of children's care. PMID- 16048080 TI - High fliers. PMID- 16048081 TI - Dangerous games. PMID- 16048082 TI - Widening the dimensions of care. PMID- 16048083 TI - Ensuring appropriate care for patients with asthma. PMID- 16048084 TI - Ankle fractures. PMID- 16048085 TI - Mass redundancies expected as SHAs tackle 750m pounds shortfall. PMID- 16048087 TI - Ward failures blamed for maternity deaths. PMID- 16048086 TI - London bombing: the response. PMID- 16048088 TI - Europe. Border line case. PMID- 16048089 TI - On gender politics. PMID- 16048090 TI - Independents' day. Interview by Helen Mooney. PMID- 16048091 TI - Data briefing. Cardiac surgery waiting times. PMID- 16048092 TI - Primary care. Otherwise engaged. AB - Professional executive committees were devised to be the clinicians' voice in primary care trusts, but staff have complained of corporate sell-out. Policies like practice-based commissioning are seen as a catalyst to reform the committees. New-style PECs are abandoning some more general work to focus on modernisation. PMID- 16048093 TI - On action research. PMID- 16048094 TI - Clinical management. Inside out. PMID- 16048095 TI - Buildings & facilities. So what's bugging you? AB - Infection control is increasingly in the media spotlight. NHS Estates is closing, causing confusion over cleanliness responsibilities. The Matron's Charter and the introduction of patient environment inspections will help clarify this. There are doubts over the assertion that one person could be held responsible for death caused by infection. PMID- 16048096 TI - Buildings & facilities. Bite of the bill. PMID- 16048097 TI - Buildings & facilities. Word power. PMID- 16048099 TI - Staff relations. Keep the faith. PMID- 16048098 TI - Buildings & facilities. A clean sweep. PMID- 16048100 TI - Fitness to work. Drink to forget. PMID- 16048101 TI - [Clinical classification of depression]. PMID- 16048102 TI - [Dementia with Lewy bodies]. AB - "Dementia with Lewy bodies (DLB)" was proposed at the first international workshop in 1995. It has received much attention since we had proposed "Lewy body disease" in 1980 and "diffuse Lewy body disease" in 1984. In the CDLB guidelines, which were reported in 1996, the clinical and pathological diagnostic criteria for DLB were shown for the first time. At present, DLB as well as Alzheimer's disease (AD) and vascular dementia (VD) are known as the three major dementing illnesses. The second international workshop was held in 1998, and the third in 2003. One of the authors, K. Kosaka, presented a paper on DLB at each international workshop, based on series of our papers which we had reported since 1976. The revised CDLB guidelines will be reported soon. In addition, the fourth international workshop on DLB will be held by Kosaka in Yokohama in 2007. In this article, we review the history, the clinical, therapeutic, neuropathological, neurochemical and molecular biological issues, based on our previous papers and other important reports on DLB. PMID- 16048103 TI - [Suicide in Tsukuba University students, 1974-2002]. AB - In this report, the author presents circumstances surrounding suicide in university students through the analysis of cases of suicide in Tsukuba University students. The subjects were 52 students (38 undergraduate, 14 graduate; 34 males, 18 females) who had committed suicide from 1974 to 2002. The average suicide rate was 18.6 per 100000, which was less than that of university students a few decades ago. The rate among liberal arts students was higher than that among science students. The number of victims increased later in the school year, and suicides among senior grades were higher. The peaks were distributed in September and March in a school years and on Tuesdays in a week. About 80% of the suicides used hanging and jumping from a height. Hanging was the most common method in both sexes, and the rate of hanging was higher in male than in female. The rate of jumping and overdose or drug was higher in female than in male. Of the 52 subjects, 16 (about 30%) had visited the outpatients clinic in the university health service center. Their clinical diagnosis was clearly divided into schizophrenic disorder and mood disorder. There was a history of suicide attempt in a half of the subjects, half of which, furthermore, had attempted suicide more than once. About half of them committed suicide less than two weeks after the last clinical visit. There might be some possible explanations for this overlooking of potential suicides, including remission of vigilance, difficulty in diagnosis and difficulty in cultivating rapport. Furthermore, suicide victims had less tendency to think of themselves as cheerful than control subjects at the point of entrance. Taking these results into consideration, the author made a few suggestions regarding suicide prevention in university students. PMID- 16048104 TI - [Present status in post-graduate psychiatry education for residencies at the general hospital]. PMID- 16048105 TI - [Present status in post-graduate psychiatry education for residencies at the general hospital without psychiatric ward]. AB - Senior psychiatrists at general hospital without psychiatric ward are usually so short of both manpower and time that they should easily lose incentives to collaborate with residents who would come at least one month under 2004 reform of residency. In this article, the author argue that the handicaps where residents should serve outpatient clinic and consultation-liaison-service at the hospital and experience other hospitals or institutions for relatively short term (mainly only one month at most) are not handicaps at all but features. Supervision during overall period of residency which concerns residents' career development and mental health, peer-supervision using live cameras or video recording, qualitative approach which utilizing weekly problem solution tasks were introduced to maintain incentives to collaboration. PMID- 16048106 TI - [Present status in post-graduate psychiatry education for residencies at the supportive-type hospital]. PMID- 16048107 TI - [On line self-assessment system for psychiatry training program]. AB - A radical reform in Japan had been done on the postgraduate clinical training system since April 2004. The subject point of the change was that in the past, mainly in the University hospitals, where the postgraduate clinical training system was made by the Department of a resident's planned medical staff, to enter in the future; in the new system, resident is incorporated in a 2-year training program with contents of internal medicine, surgery and emergency, not just bounded to particular field, in order to master basic and general clinical abilities. This new system included psychiatry rotation for 1 to 3 months. The new postgraduate clinical training system is also expected for the hospital's side to become activated by remaining the clinical training system. As a matter of course, the System has given a great effort to the curriculums of medical education. This super rotation system will be evaluated in 5-year after the introduction. The problem is whether psychiatry rotation is effective or not in a new system. If not so, psychiatry rotation might be omitted in this system. Therefore, we psychiatrists will prepare for the evidence of the effectiveness of psychiatry rotation. I developed on line self-assessment system with reference to Accreditation Council for Graduate Medical Education (ACGME) in the United States. I have introduced this system in the paper. PMID- 16048108 TI - [Proposal for new postgraduate educational rotation system in Japan]. AB - In Japan, a postgraduate educational rotation system was implemented in 2004. In this system, psychiatric training during 2-year residency is mandatory for every clinical trainee. Yokohama City University Hospital has employed a rotation system for more than 35 years, and many residents who would major in non psychiatric practice have trained in our psychiatric department. To clarify the residents' needs in psychiatric training, we carried out a questionnaire survey on our present training program. The questionnaire was returned anonymously by mail. The subjects were 43 non-psychiatric physicians who had taken a psychiatric training course in a residency training program of Yokohama City University Hospital between 1989 and 2002. The questionnaire investigated the following items; 1) reasons for choosing psychiatric training course; 2) significance of psychiatric training; 3) psychiatric disorder and psychological symptoms encountered during and after psychiatric training; 4) criticisms of the program in our hospital. We received responses from 30 of the 43 physicians surveyed. More than 90% of participants considered their experience of psychiatric training positive, and indicated that every resident should receive psychiatric training. The participants suggested that the following items are necessary; communication skill, knowledge of psychiatric disorders, and how to initiate primary care for psychiatric symptoms. These results imply that receiving psychiatric training has positive effects on the acquisition of primary care skills. However, it is important to standardize the training program in order to make the new post graduate educational rotation system more meaningful. PMID- 16048109 TI - [Needs and proposals for primary care in psychiatry]. AB - This paper focuses on primary care in psychiatry, involving both general medical primary care in psychiatric practice and psychiatric primary care in general medical practice. The author illustrates a collaborative model of care, or "Integrated Primary Care" as a possible solution for the problems in general medical primary care in psychiatry. The author describes problems in liaison between psychiatrists and general practitioners, referring to the results of the nation-wide multicenter study on pathways to psychiatric care ("Pathway Study"), which was conducted by Japan Young Psychiatrists Organization (JYPO). The author also reviews literatures on psychiatric education, and adovocates some collaborative models of mental health professionals with other medical professionals, referring to optimal educational methods of psychiatry. PMID- 16048110 TI - [Study on mental diseases: neural stem cells]. AB - The neurosciences to understand the pathophysiology of psychiatric disorders such as depression and schizophrenia were started from the analysis of neurotransmitter function change, and proceed to the cellular signaling change in the brains. In the next step, it developed to the studies of neuroplasticity change through expressions various cytokines and transcriptional factor regulations. Recently, the existence of neural stem cells and lineage of stem cells to neurons are identified in the adult mammalian brains. In addition, since the brain structural changes in depression and schizophrenia have well defined by the development of neuroimaging analysis, many researchers have been suggested the relationship between the pathophysiology of these disorders and the dysfunction of neurogenesis which strongly affects the maintenance and repair of brain neuronal network. In this report, we describe the new strategy and approach to the investigation of the treatment of psychiatric illness, and introduce our present study of the way to refine the neural network through promoted actions on stem cell functions such as differentiation to neurons. PMID- 16048111 TI - [Psycho-oncology: human side of cancer medicine]. AB - Psycho-oncology has been struggling to explore human side of cancer, which is a multidisciplinary area of oncology, epidemiology, immunology, endocrinology, sociology and bioethics, as well as psychiatry and psychology. This subspecialty addresses the two major goals; improving all aspects of quality of life, and cancer morbidity and mortality. Psycho-oncology in the 21 century addresses the needs at all stage of cancer, including prevention, diagnosis, treatment, rehabilitation, recurrence, ending active cancer treatment and palliative care. This paper focused and described current topics of psycho-oncology. PMID- 16048112 TI - [Clinical Study on sleep disorders]. PMID- 16048113 TI - Hospital chains work to reduce impact of uninsured. PMID- 16048114 TI - Acute care hospitals will be more acute. PMID- 16048115 TI - 36 million or 45 million uninsured? PMID- 16048116 TI - Activities of the USP Analytical Microbiology Expert Committee during the 2000 2005 revision cycle. AB - This article is a comprehensive review of the published activities of the Analytical Microbiology Expert Committee (AMB) for the 2000-2005 revision cycle. The major thrust of the activities during this revision cycle were directed at international harmonization, and to provide guidance in the changing field of pharmaceutical microbiology. In addition to reviewing the changes accomplished, this article discusses the rationale for many of the changes and some background information regarding new initiatives underway. Where appropriate, changes in the USP that did not fall under the direct purview of the AMB Expert Committee (EC) but of interest to the microbiology community are also discussed. PMID- 16048117 TI - "Large pore size" virus filter test method recommended by the PDA Virus Filter Task Force. PMID- 16048118 TI - Suspension for intravenous injection: image analysis of scanning electron micrographs of particles to determine size and volume. AB - The purpose of this study is to determine the size and volume of large particles in a drug suspension by performing an image analysis of digital micrographs obtained by field emission, low voltage scanning electron microscopy (FE-LVSEM). The data obtained by this method are then used to select the appropriate imaginary component of the complete refractive index necessary for the software computation of particle size distribution measured using laser light diffraction. The application of FE-LVSEM involves four major elements: 1) the use of Anodisc inorganic aluminum oxide membrane filters for the image analysis of drug crystal particles > or = 350 nm having been isolated from suspension in order to determine the area, length, width, and other particle measurements; 2) the use of either a Thornley Everhart secondary electron detector or a MCP detector in the secondary electron mode directly above the specimen so as to produce a shadow free digital image; 3) recording digital images as viewed normal to the crystal surface and at 45 degrees so as to image the edge of the crystal at a known angle; and 4) determination of drug particle volume from both views and the conversion of those volumes to an equivalent spherical volume. PMID- 16048119 TI - Permeation risks with peracetic acid and hydrogen peroxide sterilizing agent inside ambulatory pumps. AB - The sterilizing agent commonly used to sterilize materials for an isolator is a peracetic acid (PA) and hydrogen peroxide (HP) mixture. The permeation of this agent through ambulatory pumps should reveal a potential toxic risk for the patient and a stability modification of the drug by a pH change. Six wrapped and six unwrapped ambulatory pumps from each laboratory were introduced in the transfer chamber for the sterilizing process over 2 h 45 min. The presence of PA and HP were determined by using analytical strips. If the analytical strips of HP were positive, the level of HP was determined by using a specific spectrometric kit. No acid permeation was found in all wrapped pumps. Acid permeation was found in two samples of Ultraflow unwrapped series and in one unwrapped sample of Easypump series by the analytical strips. In other unwrapped samples, no acid permeation was detected. In four unwrapped ambulatory pumps (Accufuser, Infusor, Ultraflow, and Easypump), the analytical strips of HP were positive in the range of 0.5 to 25 mg/L, varying by laboratory. In only one sample (Surefuser), no detection of HP was found. The quantitative dosage of HP by spectrophotometry confirmed the permeation risk inside all pumps except the Surefuser. Our investigation shows that the permeation risk inside ambulatory pumps is real when pumps are unwrapped and exposed at high levels to PA and HP mixture. The results of our study recommend retaining the wrapping for the peracetic acid sterilization of the ambulatory pumps. PMID- 16048120 TI - Stability of all-in-one standard formulae for paediatric parenteral nutrition. AB - The Robert Debre Hospital pharmacy unit ensures an annual manufacturing rate of 20,000 parenteral nutrition bags. Until 1999, these bags consisted in binary admixtures, fat emulsions being administered to the patient via a "Y" connexion on the catheter. Since then, all-in-one standard formulae have been established and are manufactured using a Baxa MM23 automated compounder. The aim of this study was to assess the physico-chemical stability of all-in-one admixtures, in order to ensure patient administration safety (mainly avoiding precipitation risks between the nutrients). Three bags of each standard formula were manufactured in monocompartmental bags. Stability assays consisted in the assessment of the admixture's (1) macroscopic aspect, (2) drop size measurement, (3) zeta potential measurement, (4) pH measurement, and (5) osmolality measurement. Tests were conducted between D0 (manufacturing day) and D10 (10 days after manufacture). All-in-one admixtures manufactured according to the established standard formulae were found to be stable for at least 10 days, provided they are kept away from light and at a temperature of +4 degrees C. PMID- 16048121 TI - Radio-frequency identification: its potential in healthcare. AB - Radio-frequency identification (RFID) technology is just starting to make inroads into healthcare. RFID uses radio-frequency tags attached to people or objects to provide identification, tracking, security, and other functions that fall under the general heading of automatic identification and data capture (AIDC). In the retail supply chain, RFID is already well established as a way to reduce theft and track objects from manufacture through shipment to delivery. In healthcare, basic RFID is already being used to track patients for anti-elopement and anti abduction programs. As more sophisticated systems move into hospitals, RFID is also beginning to see use to provide more extensive patient identification than traditional bar coding can, and to track and locate capital equipment within the hospital. In years to come, RFID could be used for a variety of applications, including tracking and matching blood for transfusions, tracking pharmaceuticals, and combating the counterfeiting of medical products. RFID may ultimately be used for many of the functions currently carried out using bar coding--but not until the cost of RFID comes down. For the foreseeable future, the two technologies are likely to be used in tandem in many hospitals. In this article, we describe the components and operation of RFID systems and detail the different ways in which these systems are being used, and could be used, in hospitals. PMID- 16048122 TI - Keeping current with biphasic defibrillation waveforms. AB - In recent years, defibrillators that use a biphasic waveform instead of the traditional monophasic waveform have become increasingly common. In fact, since we last published on this topic in 2001, most defibrillator suppliers have begun concentrating solely on developing and marketing biphasic defibrillators while phasing out their monophasic models. Healthcare facilities are wondering if they should do the same. ECRI believes that the time has come for healthcare facilities to begin making the switch to biphasic models. That's not to say, however, that immediate replacement is required. Continuing to use a monophasic device that is in proper working condition is certainly acceptable, and replacing all models at once would be a large--and we believe unnecessary--financial burden. Rather, ECRI recommends that healthcare facilities implement a plan to phase out their monophasic defibrillators over the next few years. We discuss why, and offer recommendations to facilitate a safe and smooth transition, on the pages that follow. PMID- 16048123 TI - Limiting temperature settings on blanket and solution warming cabinets can prevent patient burns. PMID- 16048124 TI - Software conflict causes intermittent problems for Siemens CT fluoroscopy. PMID- 16048125 TI - Defective biohazard bag creates potential for bloodborne pathogen exposure. PMID- 16048126 TI - Mismeasurement using an ophthalmic ultrasound scanner results in repeat surgical procedure. PMID- 16048127 TI - Uptime guarantees for PACS--system versus component uptime. PMID- 16048128 TI - Meeting report: exploring the biology of the social amoeba Dictyostelium discoideum. international Dictyostelium conference 2004. PMID- 16048129 TI - Meeting report: evolution of protozoa and other protists, Linnean Society, London, September 13, 2004. PMID- 16048130 TI - Genetic diversity of symbiotic dinoflagellates in the genus Symbiodinium. PMID- 16048131 TI - Ultrastructural aspects of Fallisia effusa (Haemosporina: Garniidae) in thrombocytes of the lizard Neusticurus bicarinatus (Reptilia: Teiidae). AB - The fine structure of the different stages of the Fallisia effusa (Haemosporina: Garniidae), infecting the thrombocytes of the semi-aquatic Amazonian lizard Neusticurus bicarinatus (Reptilia: Teiidae) is described. Gametocytes, meronts, and merozoites of Fallisia effusa were found within a parasitophorous vacuole (PV). Multiple infections of micro- and macrogametocytes were observed. A circumferential coil of microtubules was seen in the cytoplasm of the infected host cell and this microtubule array was pronounced in cells harboring gametocytes. A deep invagination of the inner membrane complex of gametocytes may be involved in nutrition. The non-pigmented parasites underwent both merogony and gametogony in thrombocytes of the peripheral blood. No infection of the erythrocytes was observed. These observations confirm that Fallisia effusa displays characteristic features distinguishing it from other members of the Haemosporidian families, and that it has the ability to modulate microtubule assembly. PMID- 16048132 TI - The role of prey nutritional status in governing protozoan nitrogen regeneration efficiency. AB - Laboratory experiments were conducted to study nitrogen (N) regeneration by the heterotrophic marine dinoflagellate Oxyrrhis marina when ingesting phytoplankton prey of two different species and of two alternative carbon:nitrogen (C:N) ratios. Experiments were conducted in the presence of L-methionine sulfoximine (MSX) which acts as a glutamine synthetase inhibitor. Utilisation by phytoplankton of N regenerated by protozoans and other organisms drives secondary production in marine food webs. However, the rapid utilisation of this N by phytoplankton has previously hampered accurate assessment of the efficiency of protozoan N regeneration. This phenomenon is particularly problematic when the phytoplankton are nutrient stressed and most likely to rapidly utilise N. The use of MSX prevented significant utilisation by phytoplankton of protozoan regenerated N. Hence, by removing the normal pathway of N cycling, we were able to determine the N regeneration efficiency (NRE) of the protozoan. The results suggested that predator NRE could be explained in terms of the relative CN stoichiometry of prey and predator. Using a mathematical model we demonstrated that changing the method used to simulate the NRE of the protozoan trophic level has the potential to markedly modify the predicted dynamics of the simulated microbial food web. PMID- 16048133 TI - Inorganic carbon limitation and mixotrophic growth in Chlamydomonas from an acidic mining lake. AB - Plankton communities in acidic mining lakes (pH 2.5-3.3) are species-poor because they face extreme environmental conditions, e.g. 150mg l(-1) Fe2+ +Fe3+. We investigated the growth characteristics of the dominant pigmented species, the flagellate Chlamydomonas acidophila, in semi-continuous culture experiments under in situ conditions. The following hypotheses were tested: (1) Low inorganic carbon (IC) concentrations in the epilimnion (e.g. 0.3 mg l(-1)) arising from the low pH limit phototrophic growth (H-1); (2) the additional use of dissolved organic carbon (mixotrophy) leads to higher growth rates under IC-limitation (H 2), and (3) phagotrophy is not relevant (H-3). H-1 was supported as the culture experiments, in situ PAR and IC concentrations indicated that IC potentially limited phototrophic growth in the mixed surface layers. H-2 was also supported: mixotrophic growth always exceeded pure phototrophic growth even when photosynthesis was saturated. Dark growth in filtered lake water illuminated prior to inoculation provided evidence that Chlamydomonas was able to use the natural DOC. The alga did not grow on bacteria, thus confirming H-3. Chlamydomonas exhibited a remarkable resistance to starvation in the dark. The compensation light intensity (ca. 20 micromol photons m(-2) s(-1)) and the maximum phototrophic growth (1.50 d(-1)) fell within the range of algae from non acidic waters. Overall, Chlamydomonas, a typical r-strategist in circum-neutral systems, showed characteristics of a K-strategist in the stable, acidic lake environment in achieving moderate growth rates and minimizing metabolic losses. PMID- 16048134 TI - Immunocytochemical identification of the major exospore protein and three polar tube proteins of the microsporidia Paranosema grylli. AB - Microsporidia are intracellular eukaryotic parasites that can infect a wide range of animal hosts with several genera causing opportunistic infections in immunodeficient patients. Their spore wall and their unique extrusion apparatus, which has the form of a long polar tube, confer resistance of these parasites against the environment and during host-cell invasion. In contrast to parasites of vertebrates, the spore-wall and polar-tube proteins of many microsporidia species still remain to be characterized, even though a great number of microsporidia infect invertebrates. Here, we have identified one spore-wall protein and three polar-tube proteins of the microsporidia Paranosema grylli that infects the cricket Gryllus bimaculatus. Incubation of intact spores with an alkaline-saline solution resulted in the selective extraction of a major 40 kDa protein. A wash of the discharged (or destroyed) spores with SDS and the following solubilization of their polar tubes with 50-75% 2-mercaptoethanol extracted a major protein of ca. 56 kDa. When the polar tubes were solubilized in the presence of SDS, two additional proteins of 46 and 34 kDa were extracted. Antibodies specific for these extracted proteins were generated and isolated by incubation of immune sera with the protein bands that had been transferred to nitrocellulose. Western blotting demonstrated the cross-reactivity of the anti p46 and anti-p34 antibodies. Immuno-electron microscopy with the anti-p40 antibody revealed specific decoration of the microsporidia exospore. The 56, 46 and 34 kDa proteins were characterized as polar-tube components due to the clear antibody labeling of the polar filament. PMID- 16048135 TI - The identification of vahlkampfiid amoebae by ITS sequencing. AB - We have determined the internal transcribed spacer (ITS) sequences (including the 5.8S ribosomal DNA) of 30 strains of 14 species belonging to eight vahlkampfiid genera. Each previously described species has a specific ITS sequence, except for Tetramitus aberdonicus, Tetramitus thorntoni, and Tetramitus jugosus, which have identical ITS sequences. The latter three may therefore constitute a single species despite their apparent phenotypic differences. The ITS sequence appears to be conserved within a species. The species Willaertia magna appears to be ubiquitous. The 5.8S rDNA sequences of Singhamoeba horticola and Learamoeba waccamwensis indicate that they do not represent different genera, but both belong to the genus Tetramitus. The ITS sequences of 16 undescribed vahlkampfiid isolates were determined. Based on these sequences, seven isolates were identified as belonging to described species, while nine probably represent seven new species. Five of these presumed new species belong to the genus Tetramitus, and one each to the genera Vahlkampfia and Paravahlkampfia. PMID- 16048136 TI - Strains of the heterotrophic flagellate Bodo designis from different environments vary considerably with respect to salinity preference and SSU rRNA gene composition. AB - The morpho species Bodo designis is widespread and abundant globally in highly contrasting terrestrial and aquatic ecosystems. Whether the forms of Bodo designis from contrasting environments are conspecific, i.e. largely genetically identical, or whether they merely share the external morphology is presently not known. We examined the ability of different strains of Bodo designis isolated from different environments at different geographical sites to survive and grow at a salinity range of 0.5-45%. The Bodo designis strains from marine, freshwater, and terrestrial environments showed a different ability to cope with altered physiological conditions. Most of the tested strains were only able to tolerate a small salinity range, whereas others were able to withstand all tested salinity levels. We further examined the phylogenetic relationship between the different strains by sequencing the small subunit (SSU) rRNA gene. The resulting phylogenetic analyses suggest a huge genetic variation within Bodo designis, and also imply that Dimastigella and Rhyncomonas are developed inside Bodo designis. If the biological species concept is used, the genetic differences as well as the physiological barriers between the different strains of Bodo designis, would suggest that they should be assigned to different species. PMID- 16048137 TI - Vitamin D replacement therapy in persons with spinal cord injury. AB - BACKGROUND/OBJECTIVE: An increased prevalence of vitamin D deficiency has been reported in persons with chronic spinal cord injury (SCI), but treatment guidelines for replacement are not available. The purpose of this study was to evaluate two types of vitamin D therapy on calcium metabolism and vitamin D status in persons with SCI. METHODS: Ten subjects with chronic SCI who were vitamin D deficient received 25 hydroxyvitamin D3 [25(OH)D], 50 microg twice a week, for 14 days (Study 1). Regardless of vitamin D status, 40 subjects received vitamin D3 800 IU (20 microg) daily for 12 months (Study 2). Supplemental calcium was administered. The response to therapy was determined by the effect upon serum 25(OH)D levels. Results are expressed as mean +/- standard deviation. RESULTS: In Study 1, serum 25(OH)D levels increased by day 14 (8.7 +/- 2.1 vs 14.7 +/- 3.6 ng/mL; P < 0.0005). However, in 8 of 10 subjects, 25(OH)D levels were still below the absolute lower limit of normal. Serum calcium levels were not significantly different, but urinary calcium excretion increased (103 +/- 81 vs 184 +/- 145 mg/d; P < 0.01). Serum parathyroid hormone (PTH) levels decreased (35 +/- 26 vs 1 7 +/- 12 pg/ mL; P < 0.01). In Study 2, after 12 months of vitamin D supplementation, 9 subjects had an absolute and 23 had a relative vitamin D deficiency, compared with 33 and 6 subjects, respectively, at baseline. By 12 months, the 25(OH)D level increased (10.7 +/- 7.1 to 22.5 +/- 7.5 ng/mL; P < 0.0001) and the serum PTH level decreased (37 +/- 16 vs 25 +/- 11 pg/mL; P < 0.0001). CONCLUSIONS: Although 25(OH)D levels significantly increased in both studies, the replacement therapies employed were not sufficient to recommend for adoption for clinical use, indicating the need for higher doses and/or for longer periods of administration. PMID- 16048138 TI - Creatine supplementation for weak muscles in persons with chronic tetraplegia: a randomized double-blind placebo-controlled crossover trial. AB - BACKGROUND AND OBJECTIVES: Creatine supplementation improves muscle strength in some patient populations with neurologic disorders. The purpose of this study was to determine whether creatine supplementation improves muscle strength and endurance in weak upper limb muscles in persons with tetraplegia, and whether it improves function. METHODS: Outpatients with tetraplegia and mild wrist extensor weakness were randomized to receive either creatine or placebo in a double-blind crossover design. During creatine supplementation, participants were loaded with 10 g orally twice per day for 6 days, then maintained on 5 g daily until undergoing testing. Main outcome measures, performed at baseline, after placebo, and after creatine supplementation, included maximal voluntary wrist extensor isometric contraction strength (MVC), endurance times for 5 submaximal wrist extensor contractions, and the Grasp and Release Test for hand function. RESULTS: Eight individuals (7 men, 1 woman) with tetraplegia met inclusion criteria and completed all study phases. The mean age of participants was 48 years, and 7 of 8 had C6 motor level injuries. There were no significant differences in MVC, endurance times, or hand function for creatine vs placebo. CONCLUSION: Creatine does not improve MVC and endurance of weak wrist extensors and does not improve hand function in individuals with tetraplegia. PMID- 16048139 TI - Effect of fore-aft seat position on shoulder demands during wheelchair propulsion: part 1. A kinetic analysis. AB - BACKGROUND/OBJECTIVE: The highly repetitive and weight-bearing nature of wheelchair (WC) propulsion has been associated with shoulder pain among persons with spinal cord injury (SCI). Manipulation of WC seat position is believed to reduce the overall demand of WC propulsion. The objective of this investigation was to document the effect of fore-aft seat position on shoulder joint kinetics. METHODS: Thirteen men with complete motor paraplegia propelled a test WC in 2 fore-aft seat positions during free, fast, and graded conditions. The seat anterior position aligned the glenohumeral joint with the wheel axle and the seat posterior position moved the glenohumeral joint 8 cm posteriorly. The right wheel of the test chair was instrumented to measure forces applied to the pushrim. An inverse dynamics algorithm was applied to calculate shoulder joint forces, external moments, and powers. RESULTS: For all test conditions, the superior component of the shoulder joint resultant force was significantly lower in the seat-posterior position. During graded propulsion, the posterior component of the shoulder joint force was significantly higher with the seat posterior. Peak shoulder joint moments and power were similar during free and fast propulsion. During graded propulsion, the seat-posterior position displayed increased internal rotation moment, decreased sagittal plane power absorption, and increased transverse plane power generation. CONCLUSIONS: This investigation provides objective support that a posterior seat position reduces the superior component of the shoulder joint resultant force. Consequently, this intervention potentially diminishes the risk for impingement of subacromial structures. PMID- 16048140 TI - Effect of fore-aft seat position on shoulder demands during wheelchair propulsion: part 2. An electromyographic analysis. AB - BACKGROUND/OBJECTIVES: Shoulder pain is common in persons with complete spinal cord injury. Adjustment of the wheelchair-user interface has been thought to reduce shoulder demands. The purpose of this study was to quantify the effect of seat fore-aft position on shoulder muscle activity during wheelchair propulsion. METHODS: Shoulder electromyography (EMG) was recorded while 13 men with paraplegia propelled a wheelchair in the following 2 seat positions: (a) shoulder joint center aligned with the wheel axle (anterior) and (b) shoulder joint center 8 cm posterior to the wheel axle (posterior) in 3 test conditions (free, fast, and graded). Duration of EMG activity and median and peak intensities were compared. RESULTS: During free propulsion, the median EMG intensity of all muscles was similar between anterior and posterior seat positions. The major propulsive muscles (pectoralis major and anterior deltoid) demonstrated significant reductions in their median and peak intensities in the posterior seat position. Pectoralis major median intensity was significantly reduced in the posterior position during fast (52% vs 66% maximal muscle test [MMT]) and graded (41 % vs 49% MMT) conditions, and peak intensity was significantly reduced in the free condition (29% vs 52% MMT) and the fast condition (103% vs 150% MMT). Anterior deltoid intensity was significantly reduced in the posterior position during fast propulsion only (26% vs 31% MMT). For all muscles, EMG duration was similar between positions in all test conditions. CONCLUSIONS: Reduction in the intensity of the primary push phase muscles (pectoralis major and anterior deltoid) during high-demand activities of fast and graded propulsion may reduce the potential for shoulder muscle fatigue and injuries. PMID- 16048141 TI - Health literacy, morbidity, and quality of life among individuals with spinal cord injury. AB - OBJECTIVES: Health literacy has been related to indicators of health outcomes in a number of populations, but not in spinal cord injury (SCI). The current study aimed to describe levels of health literacy in SCI and to investigate its possible associations with morbidity, health-related quality of life, functional independence, community participation, and life satisfaction. DESIGN: Cross sectional survey of 107 community-living people with SCI recruited in a New Jersey outpatient SCI center. Primary measures were the Test of Functional Health Literacy in Adults (TOFHLA), standard questions about morbidity from the Behavioral Risk Factors Surveillance Study, the Craig Handicap Assessment and Reporting Technique (CHART), Short Form-12 (SF-12), and Diener's Satisfaction with Life Scale. RESULTS: Health literacy was marginal or inadequate (TOFHLA = 0 74) in only 14% of the sample. TOFHLA scores correlated significantly with physical health morbidity, even after control for severity of motor paralysis and education. American Spinal Injury Association (ASIA) Motor Scores were correlated with physical morbidity and CHART Physical Independence scores; and education was related to community Mobility, Occupation, and Economic Independence, and Satisfaction with Life scores, independent of other factors. Rasch analysis identified ceiling measurement limitations in the TOFHLA. CONCLUSIONS: Health literacy levels in this sample were higher than those found in other groups. Health literacy was independently related to physical health morbidity, but its associations with other outcomes were limited, entwined with education, and affected by severity of injury. Future research might examine higher-level aspects of health literacy to overcome ceiling measurement problems. Mediators of the association between health literacy and morbidity, such as health self-care behaviors, should also be examined. PMID- 16048142 TI - Use of a motorized bicycle exercise trainer to normalize frequency-dependent habituation of the H-reflex in spinal cord injury. AB - BACKGROUND/OBJECTIVES: Spasticity in patients with spinal cord injury (SCI) is difficult to manage. Exercise and stretching is advocated as a management tool, but these activities are difficult to perform for most patients as a result of multiple barriers. This report shows the effect of passive range-of-motion exercise in a walking-like pattern on frequency-dependent habituation of the H reflex in the lower extremities of an individual with spastic tetraplegia due to SCI. METHODS: The participant, a man with a chronic ASIA B C7 SCI due to a gunshot wound, used a motorized bicycle exercise trainer (MBET) developed at the Jackson T. Stephens Spine & Neurosciences Institute at the University of Arkansas for Medical Sciences that could be operated from the individual's wheelchair. He used the MBET for 1 hour, 5 days a week, for 13 weeks. H-reflex habituation was tested at the beginning of the study and then periodically over the course of 17 weeks, including 4 weeks after exercise had ceased. RESULTS: Significant habituation of the H-reflex was evident beginning at the 10th week of training. The habituation in the H-reflex reached a normal level at 5- and 10-Hz frequencies at 12 weeks. Subjective assessment of spasticity indicated that it was significantly reduced. The H-reflex amplitude was maintained at normal levels during the remaining week of the course of exercise and for 2 additional weeks after exercise ceased. The H-reflex habituation, however, returned to near baseline when reassessed at week 17, 4 weeks after the exercise program had concluded. Subjective assessment indicated that spasticity also had returned to pretraining levels. CONCLUSIONS: Habituation of the H-reflex, and perhaps spasticity, can be managed by a routine passive range-of-motion exercise program using a MBET, but the exercise program may need to be continuous. The benefit of reduced medication for spasticity and possibly improved quality of life could be a motivating factor for an individual with SCI and spasticity to continue the program. Because of the low complexity of the program, ease of use, and small size, this system could be inexpensive and could be used by an individual in the home. Ongoing studies will determine the minimum amount of MBET training required for maintaining long-term H-reflex habituation. PMID- 16048143 TI - A spinal cord injured animal model of lower urinary tract function: observations using direct bladder and pelvic plexus stimulation with model microstimulators. AB - BACKGROUND: Microstimulators are new devices that should be considered for management of lower urinary tract problems following spinal cord injury (SCI) such as urinary retention. These devices are small (less than 25 mm by 5 mm) with the electrodes located on the ends of the stimulator. However, it is not known whether the small electrodes on these devices would be effective in stimulating the plexus of nerves that innervate the bladder. The aim of the present study was to provide preliminary observations with model microstimulators (M-Micro) for inducing bladder contractions in an SCI animal model. Bladder wall and pelvic plexus stimulation sites were compared. Additional investigations evaluated parameters such as stimulation polarity, frequency, and period as well as bladder filling volume. METHODS: In an initial survival surgery, bilateral M-Micros were implanted on the bladder wall and the pelvic plexus along the urethra in 3 female cats. A second survival surgery was conducted 3 to 5 weeks later to produce a T1 0 SCI. Studies are reported following the second survival surgery. These studies included the effects of stimulation and bladder filling. RESULTS: The postmortem location of the implanted pelvic plexus M-Micro was previously described as near the bladder neck. Therefore, the pelvic plexus location is described in this report as "pelvic plexus (bladder neck)" stimulation. The observations showed effective stimulation with pelvic plexus (bladder neck) stimulation and voiding in some cases. Stimulation was limited by side effects of increased abdominal pressure and leg movement. Other factors also affected the response to stimulation, including the initial bladder volume and stimulating parameters. Fluoroscopy showed that when stimulation did not induce voiding the striated urethral sphincter was closed. CONCLUSIONS: This case series of 3 SCI animals showed that the small electrodes on the M-Micro could be used to stimulate the bladder with contractions and voiding in some cases. The pelvic plexus (bladder neck) location for the M-Micro may be a better location than higher on the bladder wall. Limiting side effects of stimulation included leg movement and increased abdominal pressure. Additional important factors included the stimulation parameters, initial bladder volume, and the function of the skeletal urethral sphincter. PMID- 16048144 TI - Umbilical movement after spinal cord injury. AB - BACKGROUND/OBJECTIVE: Charles Beevor first discussed the movement of the umbilicus seen upon abdominal muscle contraction in neuromuscular diseases and spinal cord injury (SCI) over a century ago. Since then, other examples of this phenomenon have been reported in the literature. METHODS: This case review study documents movements of the umbilicus caused by abdominal muscle weakness after SCI. RESULTS: SCI results in characteristic weakness of the abdominal muscles resulting in predictable movement of the umbilicus upon abdominal muscle contraction. CONCLUSIONS: Observation of umbilical movement with abdominal muscle contraction after SCI gives further confirmatory information about neurological level and type of spinal cord lesion. Further study should include information about the effect of abdominal weakness on motor and respiratory function. PMID- 16048145 TI - Respiratory management following spinal cord injury: a clinical practice guideline for health-care professionals. PMID- 16048146 TI - Nutraceutical therapies for degenerative joint diseases: a critical review. AB - There is growing recognition of the importance of nutritional factors in the maintenance of bone and joint health, and that nutritional imbalance combined with endocrine abnormalities may be involved in the pathogenesis of osteoarthritis (OA) and osteochondritis dissecans (OCD). Despite this, dietary programs have played a secondary role in the management of these connective tissue disorders. Articular cartilage is critically dependent upon the regular provision of nutrients (glucose and amino acids), vitamins (particularly vitamin C), and essential trace elements (zinc, magnesium, and copper). Therefore, dietary supplementation programs and nutraceuticals used in conjunction with non steroidal, anti-inflammatory drugs (NSAIDs) may offer significant benefits to patients with joint disorders, such as OA and OCD. This article examines the available clinical evidence for the efficacy of nutraceuticals, antioxidant vitamin C, polyphenols, essential fatty acids, and mineral cofactors in the treatment of OA and related joint disorders in humans and veterinary species. This article also attempts to clarify the current state of knowledge. It also highlights the need for additional targeted research to elucidate the changes in nutritional status and potential alterations to the expression of plasma membrane transport systems in synovial structures in pathophysiological states, so that current therapy and future treatments may be better focused. PMID- 16048147 TI - Unveiling health attitudes and creating good-for-you foods: the genomics metaphor, consumer innovative web-based technologies. AB - This article presents an integrated analysis of three emerging knowledge bases in the nutrition and consumer products industries, and how they may effect the food industry. These knowledge bases produce new vistas for corporate product development, especially with respect to those foods that are positioned as 'good for you.' Couched within the current thinking of state-of-the-art knowledge and information, this article highlights how today's thinking about accelerated product development can be introduced into the food and health industries to complement these three research areas. The 3 knowledge bases are: the genomics revolution, which has opened new insights into understanding the interactions of personal needs of individual consumers with nutritionally relevant components of the foods; the investigation of food choice by scientific studies; the development of large scale databases (mega-studies) about the consumer mind. These knowledge bases, combined with new methods to understand the consumer through research, make possible a more focused development. The confluence of trends outlined in this article provides the corporation with the beginnings of a new path to a knowledge-based, principles-grounded product-development system. The approaches hold the potential to create foods based upon people's nutritional requirements combined with their individual preferences. Integrating these emerging knowledge areas with new consumer research techniques may well reshape how the food industry develops new products to satisfy consumer needs and wants. PMID- 16048148 TI - Novel quality control methods in conjunction with chemometrics (multivariate analysis) for detecting honey authenticity. AB - The importance of honey has been recently upgraded because of its nutrient and therapeutic effect. The adulteration of honey increased exponentially in terms of both geographic and/or botanical origin. Therefore, the need has arisen for more effective quality control methods aiming at detecting adulteration. Various novel, fast, and accurate methods like AAS, HPLC, GC-MS, ES-MS, TLC, HPAED-PAD, NMR, FT-Raman, and NIR have enriched the arsenal of analytical chemist in this direction. However, apart from these novel methods, the application of multivariate analysis and, in particular, PCA, CLA, and CA, proved to be extremely useful for grouping and detecting honey of various origins. Mineral and trace element analysis were repeatedly shown to be a very effective means for the classification purposes of honey of various origins (geographical and botanical). PMID- 16048149 TI - Dietary PUFA for preterm and term infants: review of clinical studies. AB - Human milk contains n-3 and n-6 LCPUFA (long chain polyunsaturated fatty acids), which are absent from many infant formulas. During neonatal life, there is a rapid accretion of AA (arachidonic acid) and DHA (docosahexaenoic acid) in infant brain, DHA in retina and of AA in the whole body. The DHA status of breast-fed infants is higher than that of formula-fed infants when formulas do not contain LCPUFA. Studies report that visual acuity of breast-fed infants is better than that of formula-fed infants, but other studies do not find a difference. Cognitive development of breast-fed infants is generally better, but many sociocultural confounding factors may also contribute to these differences. The effect of dietary LCPUFA on FA status, immune function, visual, cognitive, and motor functions has been evaluated in preterm and term infants. Plasma and RBC FA status of infants fed formulas supplemented with both n-3 and n-6 LCPUFA was closer to the status of breast-fed infants than to that of infants fed formulas containing no LCPUFA. Adding n-3 LCPUFA to preterm-infant formulas led to initial beneficial effects on visual acuity. Few data are available on cognitive function, but it seems that in preterm infants, feeding n-3 LCPUFA improved visual attention and cognitive development compared with infants receiving no LCPUFA. Term infants need an exogenous supply of AA and DHA to achieve similar accretion of fatty acid in plasma and RBC (red blood cell) in comparison to breast-fed infants. Fewer than half of all studies have found beneficial effects of LCPUFA on visual, mental, or psychomotor functions. Improved developmental scores at 18 mo of age have been reported for infants fed both AA and DHA. Growth, body weight, and anthropometrics of preterm and term infants fed formulas providing both n-3 and n-6 LCPUFA fatty acids is similar in most studies to that of infants fed formulas containing no LCPUFA. A larger double-blind multicenter randomized study has recently demonstrated improved growth and developmental scores in a long-term feeding study of preterm infants. Collectively, the body of literature suggests that LCPUFA is important to the growth and development of infants. Thus, for preterm infants we recommend LCPUFA intakes in the range provided by feeding of human milk typical of mothers in Western countries. This range can be achieved by a combination of AA and DHA, providing an AA to DHA ratio of approximately 1.5 and a DHA content of as much as 0.4%. Preterm infants may benefit from slightly higher levels of these fatty acids than term infants. In long-term studies, feeding more than 0.2% DHA and 0.3% AA improved the status of these fatty acids for many weeks after DHA; AA was no longer present in the formula, enabling a DHA and AA status more similar to that of infants fed human milk. The addition of LCPUFA in infant formulas for term infants, with appropriate regard for quantitative and qualitative qualities, is safe and will enable the formula-fed infant to achieve the same blood LCPUFA status as that of the breast-fed infant. PMID- 16048150 TI - The importance of taking a second look at new medications. PMID- 16048151 TI - Differences between men and women regarding attitudes toward dental local anesthesia among junior students at a United Kingdom dental school. AB - The aim of this study was to assess the attitudes of junior dental students at a United Kingdom dental school regarding the administration and receipt of a local anesthetic injection. The effect of teaching on these attitudes was also evaluated. Data were collected by questionnaire. Previous experience with local anesthesia in a dental office did not affect students' attitudes toward administration and receipt of a local anesthetic injection from a classmate. Female students were more anxious about giving and receiving local anesthetic injections than male students. Didactic teaching decreased anxiety in relation to administration and receipt of a local anesthetic injection. The results showed that male and female students in a United Kingdom dental school differed in their attitudes toward local anesthesia. PMID- 16048152 TI - Midazolam premedication in children: a pilot study comparing intramuscular and intranasal administration. AB - The purpose of this study was to compare the effectiveness of intramuscular and intranasal midazolam used as a premedication before intravenous conscious sedation. Twenty-three children who were scheduled to receive dental treatment under intravenous sedation participated. The patients ranged in age from 2 to 9 years (mean age, 5.13 years) and were randomly assigned to receive a dose of 0.2 mg/kg of midazolam premedication via either intramuscular or intranasal administration. All patients received 50% nitrous oxide and 50% oxygen inhalation sedation and local anesthetic (0.2 mL of 4% prilocaine hydrochloride) before venipuncture. The sedation level, movement, and crying were evaluated at the following time points: 10 minutes after drug administration and at the times of parental separation, passive papoose board restraint, nitrous oxide nasal hood placement, local anesthetic administration, and initial venipuncture attempt. Mean ratings for the behavioral parameters of sedation level, degree of movement, and degree of crying were consistently higher but not significant in the intramuscular midazolam group at all 6 assessment points. Intramuscular midazolam was found to be statistically more effective in providing a better sedation level and less movement at the time of venipuncture than intranasal administration. Our findings indicate a tendency for intramuscular midazolam to be more effective as a premedication before intravenous sedation. PMID- 16048153 TI - Lack of pain reduction by a vibrating local anesthetic attachment: a pilot study. AB - A vibrating dental local anesthesia attachment (Vibraject, LLC, Calif) based on the concept of the gate-control theory has been used in clinical practice. The theoretical advantage of the vibrating needle is that it reduces the injection pain. We evaluated the effectiveness of Vibraject in combination with an electrical injection device. Injections were given into the alveolar mucosa adjacent to the root apex of the maxillary lateral incisor in 10 volunteers. Vibraject was randomly applied to either the left or right side of the injection. No statistically significant decrease in pain scores was found at needle insertion or anesthetic injection. The clinical efficacy of Vibraject remains controversial. PMID- 16048154 TI - A case of a power failure in the operating room. AB - In the operating room, safely administering anesthesia amidst a major power failure can instantly present one with a formidable challenge. A case is presented involving a 23-year-old healthy woman who underwent a complex oral and maxillofacial surgery to correct a dentofacial deformity. Three hours into the case and with the patient's maxilla downfractured, the overhead surgical lights blacked out, and there was an apparent loss of the anesthesia machine's ability to function. Providing adequate oxygenation, ventilation, anesthesia levels, monitoring of vital signs, and transportation of the patient were some of the challenges faced, and the response to this unexpected event is recounted. The importance of one's familiarity with an anesthesia machine's backup battery supply, routinely checking machinery, ensuring that appropriate and sufficient supplies are readily available, exercising calm leadership with clear communication, and formulating a clear plan with backup alternatives are discussed. Various recommendations are proposed with respect to the preparation for and the prevention of a power failure in the operating room. This report's account of events is aimed to "shed some light" on this topic, serve as a check of one's own preparedness, and facilitate the optimal management of a similarly unexpected incident. PMID- 16048155 TI - Outpatient anesthesia for oral surgery in a juvenile with Leigh disease. AB - We report a case of anesthesia for elective outpatient third molar extraction in a juvenile with Leigh disease, a progressive neurodegenerative disorder related to respiratory chain deficiency. This syndrome usually presents in infancy and is characterized by nervous system dysfunction and respiratory abnormalities. Anesthesia has been reported to aggravate respiratory symptoms and frequently precipitate respiratory failure. Preoperative swallowing difficulty or respiratory symptoms should be carefully diagnosed, because they can be a warning sign of postoperative complications or mortality. Adverse effects of anesthesia may quickly lead into metabolic acidosis. Anesthetics should be carefully chosen that do not interfere with mitochondrial respiration, which can lead to lactic acidosis. PMID- 16048156 TI - Specialist shortage shakes emergency rooms; more hospitals forced to pay for specialist care. PMID- 16048157 TI - ACPE poll: physician leaders distressed by specialist shortage; on call pay controversial. PMID- 16048158 TI - 100,000 lives campaign adds 1,700 hospitals...and counting. PMID- 16048159 TI - Defeating the malpractice crisis with standardized review of expert opinions. PMID- 16048160 TI - How to successfully dissolve a hospital-owned practice. PMID- 16048161 TI - Health care leaders as agents of change. PMID- 16048162 TI - ABCs of TJR--physician involvement helps build a profitable program. PMID- 16048163 TI - An overview of the drug development process. PMID- 16048164 TI - Finding an incentive plan that actually works. PMID- 16048165 TI - Closing quality and value gaps (part 3). PMID- 16048166 TI - Rebuilding the ordinary into the extraordinary. PMID- 16048167 TI - Consumer directed health care: zeroing in on physician practices. PMID- 16048168 TI - Two recent opinions allow hospitals to assist independent physicians. PMID- 16048169 TI - Using value innovation to create competitive advantage (part I). PMID- 16048170 TI - Following a proven path to success. PMID- 16048171 TI - Thinking about thinking. PMID- 16048172 TI - The development of the quartz crystal oscillator industry of World War II. AB - This paper offers a history of a critical episode in military and electronics history-the difficult creation of quartz crystal frequency control units for radio communications during World War II. As a means of controlling the frequencies of radio transmitters and receivers, amateur radio hobbyists quickly accepted the quartz crystal oscillator after its initial development in the late 1920s. The military, however, declined to adopt this technology until just prior to World War II. Due to the small market for crystal oscillators, no mass production industry had ever developed to produce this extremely high precision electronic component. As war engulfed the nation, the U.S. Army Signal Corps found itself in the dangerous position of having gambled the integrity of its communications equipment on a component that could not possibly be produced in the quantities immediately needed. This paper looks at the challenges the United States faced in building a crystal manufacturing capability and in supplying this industry with sufficient supplies of raw quartz. A fairly specialized component of communications technology emerged from spare beginnings in prewar amateur radio to become the very foundation of a wide range of electronic devices today. PMID- 16048173 TI - A review of analyses related to vibrations of rotating piezoelectric bodies and gyroscopes. AB - A review of analyses on vibrations of rotating piezoelectric structures for applications in piezoelectric angular rate sensors is presented. The operating principle and the basic behaviors of piezoelectric gyroscopes are shown by a simple example. Structural shapes and vibration modes that can be used to make gyroscopes are summarized. Analytical and numerical results from the three dimensional equations of a rotating piezoelectric body and the corresponding one- and two-dimensional theories of piezoelectric beams, rings, plates, and shells are reviewed. PMID- 16048174 TI - Thin film resonator technology. AB - Advances in wireless systems have placed increased demands on high performance frequency control devices for operation into the microwave range. With spectrum crowding, high bandwidth requirements, miniaturization, and low cost requirements as a background, the thin film resonator technology has evolved into the mainstream of applications. This technology has been under development for over 40 years in one form or another, but it required significant advances in integrated circuit processing to reach microwave frequencies and practical manufacturing for high-volume applications. This paper will survey the development of the thin film resonator technology and describe the core elements that give rise to resonators and filters for today's high performance wireless applications. PMID- 16048175 TI - Progress in medical ultrasound exposimetry. AB - Biomedical applications of ultrasound have experienced tremendous growth over the past 50 years. Early work in thermal therapy and surgery soon was followed by diagnostic imaging and Doppler. Because patient safety was an important issue from the beginning, the study of methods for measuring exposure levels, and their relationship to possible biological effects, paralleled the growth of the various therapeutic and diagnostic techniques. The diverse conditions of use have presented a range of exposure measurement challenges, and the sensors and techniques used to evaluate ultrasound fields have had to evolve as new or expanded clinical applications have emerged. In this paper some of the more notable of these developments are presented and discussed. Topics covered include devices and techniques, methods of calibration, progress in standardization, and current problem areas, including the effects of nonlinear propagation. Some early methods are described, but emphasis is given to more recent work applicable to present and future uses of ultrasound in medicine and biology. PMID- 16048176 TI - The piezoelectric semiconductor and acoustoelectronic device development in the sixties. AB - In the 1960s the properties of piezoelectric semiconductors, group III-V zinc blende and group II-VI wurtzite structure, were explored for the development of acoustoelectronic (AE), devices. Bulk acoustic wave (BAW), delay lines, traveling wave amplifiers, and oscillators were developed. Although these elegant functional devices never made it into the realm of full-scale production and application, the piezoelectric semiconductor developments of the 1960s provided an exciting time for theoretical explanations and creative experimentation to explore device capabilities for electronic systems applications. Delay lines were formed from rectangular parallelepiped blocks of piezoelectric semiconductors with integral input and output transducers depleted of carriers at each end of the block. The ultrasonic traveling wave amplifier was based on the interaction of electrons under a bias field moving with a velocity faster than the piezoelectric field accompanying the acoustic traveling wave. The gain factor in a piezoelectric semiconductor under direct current bias was used to develop oscillators. The main research efforts were carried out by industrial laboratories with government funding support. This paper considers the work with piezoelectric semiconductors during the 1960s with examples from the author's own work. PMID- 16048177 TI - Piezoelectric composites for sensor and actuator applications. AB - In the last 25 years, piezoelectric ceramic-polymer composites have been conceptualized, prototyped, fabricated, and implemented in an array of applications encompassing medical imaging and military missions, among others. A detailed snapshot of the materials used, and a detailed account of the major innovative methods developed in making various piezoelectric ceramic-polymer composites are presented. The salient aspects of processing of such composites are summarized, and structure-processing-property relations are described using connectivity as the unifying central concept. Computer-aided design (CAD)-based fabrication methods, which result in composites whose structural complexity surpass that of composites obtained with traditional methods, are described to introduce the reader to novel concepts in processing of piezocomposites. A brief survey of some recent advances made in modeling of (0-3), (1-3), and (2-2) composites also is provided. PMID- 16048178 TI - The history and properties of ultrasonic inhomogeneous waves. AB - This paper gives a historical survey of the development of the inhomogeneous wave theory, and its applications, in the field of ultrasonics. The references are listed predominantly chronologically and are as good as complete. Along the historical description, several scientific features of inhomogeneous waves are described. All topics of inhomogeneous wave research are taken into account, such as waves in viscoelastic solids and liquids, thermoviscous liquids and solids, and anisotropic viscoelastic materials. Also inhomogeneous waves having complex frequency are described. Furthermore, the formation of bounded beams by means of inhomogeneous waves is given and the diffraction of inhomogeneous waves on periodically corrugated surfaces. The experimental generation of inhomogeneous waves is considered as well. PMID- 16048179 TI - Acoustic waves in bounded anisotropic media: theorems, estimations, and computations. AB - This paper discusses some basic achievements in theoretical studies on acoustic wave propagation along boundaries in anisotropic solids. In particular, the following issues are reviewed: existence theorems for subsonic surface and interface waves, leaky surface acoustic waves (SAW) and their relation to "supersonic" SAWs and fast exceptional bulk waves, the resonance reflection of bulk waves in the vicinity of leaky wave branches. General conclusions are illustrated by numerical examples. PMID- 16048180 TI - The radiation mode theory in ultrasonics. AB - This paper describes the history and the state of the art in radiation mode theory (RMT) in ultrasonics. The RMT originates from electromagnetism in which it has proved to be very efficient in the field of wave guides and discontinuities. In ultrasonics, the RMT made its entrance only a decade ago and has already proved to be very efficient in describing the interaction of sound with discontinuities such as a step on a plate, a liquid wedge, the extremity of a plate and much more. It is likely that the development of the RMT for two dimensional (2-D) isotropic media has come almost to an end. This paper lists the results obtained so far. Further extensions to more complicated media are to be expected in the coming decade. PMID- 16048181 TI - Shear horizontal BG surface acoustic waves on piezoelectrics: a historical note. AB - In December of 1968 Jeffrey Bleustein of Yale University published an article predicting the existence of a new type of transverse surface acoustic wave (SAW) that could propagate on the surface of a piezoelectric crystal. This was followed within 20 days by the publication of an article by Yuri Gulyaev in January of 1969 predicting the same basic surface wave propagation. The wave took on the name Bleustein-Gulyaev or BG-wave, joining the names of Rayleigh, Love, Sezawa, and Stonely for distinct types of SAW. But is there more to the story than this? Were there works preceding those of Bleustein and Gulyaev which signaled an interest in exploring a new surface wave mode on piezoelectrics? What about the work of Shimizu, Nakamura, and Ohta, who in April of 1969 published both theoretical and experimental verification of the existence of such a wave independent of the knowledge of the Bleustein and Gulyaev papers? This paper explores the early roots and characteristics of the shear horizontal surface wave on a piezoelectric. PMID- 16048182 TI - Surface transverse waves: properties, devices, and analysis. AB - Surface transverse waves represent a new generation of the surface acoustic wave (SAW) family that offers advantageous properties without further demand for new materials or improved design and technology. The most effective activity in the surface transverse wave (STW) area has been realized during the last decade with high-performance devices achieved and analytical methods developed. The present paper reviews the basic achievements in historical and factual order. A state-of the-art introduction is combined with discussion on the development tendencies with specific emphasis on sensor technology. PMID- 16048183 TI - Causality-imposed (Kramers-Kronig) relationships between attenuation and dispersion. AB - Causality imposes restrictions on both the time-domain and frequency-domain responses of a system. The Kramers-Kronig (K-K) relations relate the real and imaginary parts of the frequency-domain response. In ultrasonics, K-K relations often are used to link attenuation and dispersion. We review both integral and differential forms of the frequency-domain K-K relations that are relevant to theoretical models and laboratory measurements. We consider two methods for implementing integral K-K relations for the case of finite-bandwidth data, namely, extrapolation of data and restriction of integration limits. For the latter approach, we discuss the accuracy of K-K predictions for specific classes of system behavior and how the truncation of the integrals affects this accuracy. We demonstrate the accurate prediction of attenuation and dispersion using several forms of the K-K relations relevant to experimental measurements of media with attenuation coefficients obeying a frequency power law and media consisting of resonant scatterers. We also review the time-causal relations that describe the time-domain consequences of causality in the wave equation. These relations can be thought of as time-domain analogs of the (frequency-domain) K-K relations. Causality-imposed relations, such as the K-K and time-causal relations, provide useful tools for the analysis of measurements and models of acoustic systems. PMID- 16048184 TI - An automatic tracking system for phase-noise measurement. AB - A low cost, automatic tracking system for phase noise measurement has been implemented successfully. The tracking system is accomplished by applying a charge pump phase-locked loop as an external reference source to a digital spectrum analyzer. Measurement of a 2.5 GHz, free-running, voltage-controlled oscillator demonstrated the tracking accuracy, thus verifying the feasibility of the system. PMID- 16048185 TI - Polarization-switching D/A converter. AB - This paper describes a novel digital-to-analog (D/A) conversion technique, which uses the analog quantity polarization as a D/A conversion medium. It can be implemented by CMOS capacitors or by ferroelectric capacitors, which exhibit strong nonlinearity in charge versus voltage behavior. Because a ferroelectric material inherently has spontaneous polarization and generally has a large dielectric constant, the effective capacitance of a ferroelectric capacitor is much larger than that of a CMOS capacitor of the same size. This ensures less influence of bottom-electrode parasitic capacitance on a ferroelectric capacitor. Furthermore, a data converter based on ferroelectric capacitors possesses the potential nonvolatile memory function owing to ferroelectric hysteresis. Along with the architecture proposed for polarization-switching digital-to-analog converter (PDAC), its circuit implementation is introduced. Described is implementation of two 9-bit bipolar PDACs: one is based on CMOS capacitors and the other on off-chip ferroelectric capacitors. Experimental results are presented for the performance of these two prototypes. PMID- 16048186 TI - Identification of vulnerable atherosclerotic plaque using IVUS-based thermal strain imaging. AB - Pathology and autopsy studies have demonstrated that sudden disruption of vulnerable atherosclerotic plaque is responsible for most acute coronary syndromes. These plaques are characterized by a lipid-rich core with abundant inflammatory cells and a thin fibrous cap. Thermal strain imaging (TSI) using intravascular ultrasound (IVUS) has been proposed for high-risk arterial plaque detection, in which image contrast results from the temperature dependence of sound speed. It has the potential to distinguish a lipid-laden lesion from the arterial vascular wall due to its strong contrast between water-bearing and lipid bearing tissue. Initial simulations indicate plaque identification is possible for a 1 degrees C temperature rise. A phantom experiment using an IVUS imaging array further supports the concept, and results agree reasonably well with prediction. PMID- 16048187 TI - Statistical estimation of ultrasonic propagation path parameters for aberration correction. AB - Parameters in a linear filter model for ultrasonic propagation are found using statistical estimation. The model uses an inhomogeneous-medium Green's function that is decomposed into a homogeneous-transmission term and a path-dependent aberration term. Power and cross-power spectra of random-medium scattering are estimated over the frequency band of the transmit-receive system by using closely situated scattering volumes. The frequency-domain magnitude of the aberration is obtained from a normalization of the power spectrum. The corresponding phase is reconstructed from cross-power spectra of subaperture signals at adjacent receive positions by a recursion. The subapertures constrain the receive sensitivity pattern to eliminate measurement system phase contributions. The recursion uses a Laplacian-based algorithm to obtain phase from phase differences. Pulse-echo waveforms were acquired from a point reflector and a tissue-like scattering phantom through a tissue-mimicking aberration path from neighboring volumes having essentially the same aberration path. Propagation path aberration parameters calculated from the measurements of random scattering through the aberration phantom agree with corresponding parameters calculated for the same aberrator and array position by using echoes from the point reflector. The results indicate the approach describes, in addition to time shifts, waveform amplitude and shape changes produced by propagation through distributed aberration under realistic conditions. PMID- 16048188 TI - Compact FPGA-based beamformer using oversampled 1-bit A/D converters. AB - A compact medical ultrasound beamformer architecture that uses oversampled 1-bit analog-to-digital (A/D) converters is presented. Sparse sample processing is used, as the echo signal for the image lines is reconstructed in 512 equidistant focal points along the line through its in-phase and quadrature components. That information is sufficient for presenting a B-mode image and creating a color flow map. The high sampling rate provides the necessary delay resolution for the focusing. The low channel data width (1-bit) makes it possible to construct a compact beamformer logic. The signal reconstruction is done using finite impulse reponse (FIR) filters, applied on selected bit sequences of the delta-sigma modulator output stream. The approach allows for a multichannel beamformer to fit in a single field programmable gate array (FPGA) device. A 32-channel beamformer is estimated to occupy 50% of the available logic resources in a commercially available mid-range FPGA, and to be able to operate at 129 MHz. Simulation of the architecture at 140 MHz provides images with a dynamic range approaching 60 dB for an excitation frequency of 3 MHz. PMID- 16048189 TI - Ultrasound research scanner for real-time synthetic aperture data acquisition. AB - Conventional ultrasound systems acquire ultrasound data sequentially one image line at a time. The architecture of these systems is therefore also sequential in nature and processes most of the data in a sequential pipeline. This often makes it difficult to implement radically different imaging strategies on the platforms and makes the scanners less accessible for research purposes. A system designed for imaging research flexibility is the prime concern. The possibility of sending out arbitrary signals and the storage of data from all transducer elements for 5 to 10 seconds allows clinical evaluation of synthetic aperture and 3D imaging. This paper describes a real-time system specifically designed for research purposes. The system can acquire multichannel data in real-time from multi element ultrasound transducers, and can perform some real-time processing on the acquired data. The system is capable of performing real-time beamforming for conventional imaging methods using linear, phased, and convex arrays. Image acquisition modes can be intermixed, and this makes it possible to perform initial trials in a clinical environment with new imaging modalities for synthetic aperture imaging, 2D and 3D B-mode, and velocity imaging using advanced coded emissions. The system can be used with 128-element transducers and can excite 128 transducer elements and receive and sample data from 64 channels simultaneously at 40 MHz with 12-bit precision. Two-to-one multiplexing in receive can be used to cover 128 receive channels. Data can be beamformed in real time using the system's 80 signal processing units, or it can be stored directly in RAM. The system has 16 Gbytes RAM and can, thus, store more than 3.4 seconds of multichannel data. It is fully software programmable and its signal processing units can also be reconfigured under software control. The control of the system is done over a 100-Mbits/s Ethernet using C and Matlab. Programs for doing, e.g., B-mode imaging can be written directly in Matlab and executed on the system over the net from any workstation running Matlab. The overall system concept is presented along with its implementation and examples of B-mode and in vivo synthetic aperture flow imaging. PMID- 16048190 TI - Ultrasonic guided wave scattering in a plate overlap. AB - Guided wave scattering in a plate overlap is investigated by numerical calculations and experimental measurements of transmission and reflection factors from the overlap region. In the numerical study, a hybrid boundary element-finite element method is used to calculate the guided wave scattered field from the overlap region. Transmission and reflection factors are calculated for incident A0 and S0 Lamb and n0 shear horizontal waves, including higher modes generated through mode conversion phenomena. In addition, parametric studies of transmission and reflection factors in this problem are performed numerically over various incident modes, frequencies, and overlap lengths. For verification and comparison with numerical results, experiments were conducted to measure the transmission and reflection factors for incident Lamb and shear horizontal waves in steel plates with two different overlap areas. The experimental results agree well with the numerical calculations. The numerical and experimental results show that it is highly feasible to carry out efficient Lamb wave nondestructive evaluation (NDE) in overlapped plates and in multilayer structures with various lap joints by selecting various modes and tuning frequency. PMID- 16048191 TI - Resonance reflection of acoustic waves in piezoelectric bi-crystalline structures. AB - The paper studies the bulk wave reflection from internal interfaces in piezoelectric media. The interfaces of two types have been considered. Infinitesimally thin metallic layer inserted into homogeneous piezoelectric crystal of arbitrary symmetry. Rigidly bonded crystals whose piezoelectric coefficients differ by sign but the other material constants are identical. Analytic expressions for the coefficients of mode conversion have been derived. An analysis has been carried out of specific singularities arising when the angle of incidence is such that the resonance excitation of leaky interface acoustic waves occurs. The conditions for the resonance total reflection have been established. The computations performed for lithium niobate (LiNbO3) illustrate general conclusions. PMID- 16048192 TI - Investigation of high power effects on Ti/Al and Ta-Si-N/Cu/Ta-Si-N electrodes for SAW devices. AB - Damage behavior of two different metallization systems (Ti/Al bilayer and Ta-Si N/Cu/Ta-Si-N multilayer) as finger electrodes in surface acoustic waves (SAW) devices was investigated. A special test structure was developed for this reason. The samples were loaded with traveling SAWs varying input power and loading time. Simultaneously during these experiments, the electric behavior of the SAW structure was measured and damage development by voids and hillock formation was observed using optical microscopy, too. The damaged structures were investigated by means of different microscopy techniques. Results show that the Cu-based metallization system has a significantly higher acoustomigration resistance and power durability in comparison with the Al thin film system. PMID- 16048193 TI - Thickness-shear vibration of rotated Y-cut quartz plates with relatively thick electrodes of unequal thickness. AB - An exact solution is obtained from the three-dimensional equations of linear piezoelectricity for thickness-shear vibrations of rotated Y-cut quartz plates with relatively thick electrodes of unequal thickness at its major faces. Effects of the shear stiffness of the electrodes on resonant frequencies are examined. PMID- 16048194 TI - Investigation of layered structure SAW devices fabricated using low temperature grown AlN thin film on GaN/sapphire. AB - Epitaxial AlN films have been grown on GaN/sapphire using helicon sputtering at 300 degrees C. The surface acoustic wave (SAW) filters fabricated on AlN/GaN/sapphire exhibit more superior characteristics than those made on GaN/sapphire. This composite structure of AlN on GaN may bring about the development of high-frequency components, which integrate and use their semiconducting, optoelectronic, and piezoelectric properties. PMID- 16048195 TI - Triumph over adversity. PMID- 16048196 TI - Are directors of nursing being eclipsed by the modern matron? PMID- 16048197 TI - Bedtime blues. Helping your children get a good night's sleep. AB - All new parents expect some broken nights but when a child's wakefulness becomes excessive it can lead to exhaustion and even marital breakdown. Sleep counsellors can help but are often in short supply. PMID- 16048198 TI - Global virus alert. How SARS highlighted the risk of viral pandemics. AB - As recent events involving SARS and bird flu have shown, foreign travel can be a hazardous undertaking. The burden of disease is high with between 20 and 70 per cent of the 50 million people visiting developing countries reporting a travel associated illness. PMID- 16048199 TI - The new truth. Why nurses are playing a bigger role in spiritual care. PMID- 16048200 TI - A language barrier isolates patients and can be harmful. PMID- 16048201 TI - Everyone's business. Infection control in Scotland is now public property. PMID- 16048202 TI - Easing the pain. Should cannabis be used for chronic pain relief? PMID- 16048203 TI - Early discharge: give it the push. PMID- 16048204 TI - The big squeeze. PMID- 16048205 TI - Understanding the lives of older women. AB - AIM: To understand why some women adjust successfully in later life while others, in similar circumstances, become overwhelmed by life and dislike their old age. METHOD: Qualitative data were generated from field notes of participant observation carried out over ten months. In-depth interviews were also carried out with 11 young older and frail older women, aged 60 to 89 years, attending day centres. A grounded theory approach was adopted to collect and analyse participant observation and interview data. FINDINGS: Adjustment in later life occurs regardless of circumstance. Three areas were identified as being the main obstacles for many. These are depression, maintaining intimacy through friends and family, and managing the change process associated with older age. In addition, a core category of searching for meaning was discovered and used to understand how depression, intimacy and managing change affect adjustment in later life. CONCLUSION: Nurses could use their skills and experience of listening carefully to older patients to discover whether they are distressed, and assess whether there is a need for psychological and emotional support. There is a need for more research into carrying out adjustment work with older women who are struggling to enjoy their later years and with older men to discover themes that are relevant to them. PMID- 16048206 TI - Reflections of a novice researcher. AB - The aim of this article is to highlight the experiences of a novice nurse researcher from a biographical point of view. The article describes how nurse education in Australia has incorporated nursing research and discusses the issue of integrating research into practice. PMID- 16048207 TI - Dietary guidelines for the management of diabetes. AB - This article outlines contemporary guidelines for the dietary management of diabetes, with the aim of reducing inconsistencies in dietary recommendations. PMID- 16048208 TI - Diagnosing the ache. PMID- 16048209 TI - Change of tack. PMID- 16048210 TI - Magic moments. The rewards of good management make nursing worthwhile. PMID- 16048211 TI - One year too many. Proposals to introduce a probationary year for newly qualified students need close inspection. PMID- 16048212 TI - Challenging learners to think critically. PMID- 16048214 TI - Benefits of surgical reconstruction in pressure ulcers with a non-advancing edge and scar formation. AB - OBJECTIVE: To highlight the difficulty in treating pressure ulcers associated with scar formation and the benefits of surgical treatment in these cases by presenting a series of clinical cases. METHOD: Nine sacral ulcers surrounded by significant scar formation and 14 sacral ulcers without significant scars were treated using the same conservative management protocol. Healing status was quantitatively estimated using an assessment tool called DESIGN, which classifies pressure ulcer severity and allows healing to be monitored; a decreasing score indicates progressive improvement. RESULTS: Ulcers with scar formation showed little response to the treatment, while those without remarkable scars exhibited a significant decrease of the DESIGN scores. Surgical reconstruction was conducted in five ulcers surrounded by scar formation, and all showed complete healing within extremely short periods. CONCLUSION: Conservative treatment of pressure ulcers with a non-advancing edge surrounded by scar formation presents considerable difficulties. Surgical reconstruction offers many benefits for non healing ulcers. PMID- 16048215 TI - Redesigning the process of providing compression hosiery to patients attending hospital clinics. PMID- 16048216 TI - Cutaneous calciphylaxis in a haemodialysis patient. PMID- 16048217 TI - Wounds scrutiny in a Swedish hospital: prevalence, nursing care and bacteriology, including MRSA. AB - OBJECTIVE: To identify all wounds, wound types and wound characteristics; to identify bacteria in all wounds, particularly MRSA, VRE and multi-resistant Gram negative rods;. METHOD: All patients admitted to or visiting hospital clinics were examined. If the patient had a wound, a questionnaire was filled in by a nurse, and if the wound met the inclusion criteria swabs were taken. RESULTS: A total of 2172 patients were admitted to or visited the hospital; 408 (19%) had a total of 668 wounds. Of this number, 248 were cultured. Thirty-seven patients had pressure ulcers, 83 had leg and foot ulcers and 288 patients had other types of wounds. Nursing care varied according to wound type and ward. Fifty-eight different types of wound dressings were used. Cleansing was performed with saline in 58% of the wounds. The mean number of dressing changes was once daily. Wounds were painful in 37% of the cases, with a predominance of leg and foot ulcers (51%). Many patients did not receive analgesia. MRSA was identified in two patients. No VRE was identified, and there was a low prevalence of multi resistance in Enterobacteriaceae and Pseudomonas aeruginosa. CONCLUSION: The study provided important information for future improvement of wound care in a university hospital. Fewer resistant bacteria, particularly MRSA, were identified than expected. PMID- 16048218 TI - Enzymatic versus autolytic debridement of chronic leg ulcers: a prospective randomised trial. AB - OBJECTIVE: A randomised clinical trial (n = 42) compared the effectiveness of two approaches to debriding chronic leg ulcers: TenderWet 24, which is designed to support the autolytic degradation process, and Iruxol N (Santyl), an enzymatic treatment claimed to enhance the degradation process. METHOD: Patients were randomly assigned to one of the two treatment groups for three weeks. Wounds were evaluated weekly for the amount of eschar/slough, the area of healthy granulation and the re-epithelialised area. RESULTS: During days 1-14 slough within the groups was reduced by almost 19% for TenderWet 24 and by 9% for Iruxol N, followed by an increase of 26% and 10% respectively in granulation tissue. These effects were less accentuated during days 7-21. There was a further 11% improvement in tissue debridement for the TenderWet 24 group and a relapse (+9.1%) in the Iruxol N group. CONCLUSION: Although TenderWet 24 appeared to be more efficient in a few cases, the general efficacy of the two products appeared to be almost the same as no statistically significant superiority of either product was found. PMID- 16048219 TI - The role of the fibrin cuff in the development of venous leg ulcers. PMID- 16048220 TI - Evaluation of a lipidocolloid wound dressing in the local management of leg ulcers. AB - OBJECTIVE: To evaluate the efficacy, tolerance and acceptability of Urgotul and DuoDERM E dressings in the local management of venous or mixed-aetiology leg ulcers. METHOD: This was a prospective multicentre randomised phase IV clinical trial conducted open-label in parallel groups. It involved 20 investigating centres, including hospital dermatology and vascular medicine departments, and private practices. Dermatologists and angiologists/phlebologists took part. Subjects were adult, non-immunosuppressed patients presenting with a non infected, non-malignant leg ulcer of predominantly venous origin (ABPI > 0.8). Ulcers were between 4cm2 and 40cm2 in size, with granulation tissue covering more than 50% of their surface area. Ulcer duration ranged from three to 18 months. Patients were followed-up by the investigating physician for eight weeks on a weekly basis; this included clinical examination, wound area tracings and photographs. Nurses (hospital or visiting) assessed exudate volume and clinical appearance at dressing changes. RESULTS: Ninety-one patients were included: 47 in the Urgotul group and 44 in the DuoDERM E group. Baseline patient demographic data and wound characteristics were comparable in the two groups. After eight weeks of treatment wound surface area had reduced by a mean of 61.3% in the Urgotul group and 52.1% in the DuoDERM E group (NS); dressings were changed more frequently in the DuoDERM E group (2.54 +/- 0.57 times per week versus 2.31 +/- 0.45 in the Urgotul group, p = 0.047). Thirty-three local adverse events were recorded in 27 patients: 10 in the Urgotul group and 23 in the DuoDERM E group (p = 0.039). Nurses reported better acceptability for the Urgotul dressing, based on pain on removal, maceration and odour (p < 0.0001). CONCLUSION: Both dressings showed similar efficacy for the local treatment of venous leg ulcers. Nevertheless, medical and nursing staff reported better tolerance and acceptability for the Urgotul dressing. PMID- 16048221 TI - Leg wound infections following cardiac surgery: a scoring system for assessment and management. AB - OBJECTIVE: Well-defined criteria are needed to provide guidance for the appropriate management of leg wounds following saphenous vein harvest in coronary artery bypass graft surgery (CABG). METHOD: A score named DISINFECT was devised to carefully define the variables to be considered for assessing saphenous vein harvest wounds. RESULTS: This preliminary study included 100 consecutive patients undergoing first-time isolated CABG requiring the saphenous vein as a conduit. Wounds were assessed and the points combined to create a daily score (D) according to the presence of increased C-reactive protein/white blood cells (I), surrounding tissue (S), quality of the incision (I), new skin (N), foreign material (F), exudate (E), positive cultures (C) and temperature (T). CONCLUSION: Taking into account the stages of wound healing, severity of infection and appropriate use of antibiotics, this method of wound management would improve the consistency with which leg wounds are managed, reduce hospital stays and increase resistance to hospital-acquired infection. PMID- 16048222 TI - Looking up. Credit rating upgrades again outpace downgrades. PMID- 16048223 TI - Medicaid panel chosen. PMID- 16048224 TI - Scrushy plots a comeback. As HealthSouth works hard to put a fraud scandal in the past, its acquitted founder studies how he can regain the company. AB - The jury ruled that Richard Scrushy is not guilty. So now HealthSouth's deposed leader wants to reclaim the company he founded, even if current management and investors don't want him. "I have a hard time believing he's going to get back in there," says Larry Scanlan, left, of Navigant Consulting. Others are more blunt: "We have less than zero support for his return," says a principal at the largest single owner of HealthSouth shares. PMID- 16048225 TI - New York gets rattled. St. Vincent's bankruptcy puts local hospitals on edge. PMID- 16048226 TI - Sending out shock waves. UnitedHealth to add PacifiCare's Medicare market. PMID- 16048227 TI - High stakes in Evanston case. If FTC loses, hospital merger enforcement may lag. PMID- 16048228 TI - Nurse agency sues state group. PMID- 16048229 TI - Battle in Alabama. Agency, system fight for control of Cullman hospital. PMID- 16048230 TI - Accreditation complication. Four Lovelace Sandia hospitals denied JCAHO nod. PMID- 16048231 TI - Calculating what it's worth. Value analysis pros making mark in healthcare. PMID- 16048232 TI - Vista wins important convert. PMID- 16048233 TI - Lessons from the debacle. The Scrushy verdict is hardly the end of enforcing corporate accountability. PMID- 16048234 TI - Not dead yet. New technology has not completely eclipsed the medical transcription industry, but its many problems put the industry at risk. PMID- 16048235 TI - Lost in translation. Many errors traced to physicians' sloppy speech. PMID- 16048236 TI - By the number. Largest healthcare executive search firms ranked by 2004 U.S. retained healthcare services executive search consulting fee revenue. PMID- 16048237 TI - Taking measure. New study quantifies problem of adverse drug events. PMID- 16048238 TI - Multivariate analysis in suicide attempts: a useful tool for the clinician. PMID- 16048239 TI - Brief communication: factors affecting rehospitalisation in psychiatric patients in Singapore. AB - OBJECTIVES: To study the patterns of readmission in Singapore using a cohort of first admission schizophrenia patients to the state mental health institute. METHODS: A retrospective analysis of case records was done of first admission schizophrenia patients to the hospital from April 1993 to March 1994. Patients who had only one admission in the five years of follow up were compared to 'revolving door patients'. RESULTS: The cohort comprised 133 males (56.8%) and 101 females (43.2%). Seventy-eight patients (33.3%) had one admission to the hospital while 56 (23.9%) fulfilled the revolving door criteria. On performing a logistic regression we found sex (p = 0.02), shorter duration of illness (p = 0.01) and CPN referral (0.006) were significantly associated with revolving door phenomena. CONCLUSIONS: Our study has identified male gender, history of self harm and a short duration of illness to be risk factors for 'revolving door' phenomena. PMID- 16048240 TI - An historical review of the mental health services in the People's Republic of China. AB - BACKGROUND: This paper is an attempt to describe the historical development of the mental health services in the PRC. RESEARCH METHODS: An archive of related literature. FINDINGS: The development of the mental health services in the PRC could be divided into several stages: the introduction of mental asylums by western missionaries before 1949; indigenization of the treatment model after the establishment of the PRC (1949-1963). Strong political control governed diagnosis and treatment as well as detention and discharge of mental patients during the Cultural Revolution (1964-1976). Later, because of modernization and reform advocated by Deng Xiao-ping, western models of treatment and rehabilitation were gradually introduced by psychiatrists in the PRC. Nevertheless, dilemmas such as human rights versus political control, community integration versus community control, diversity versus centrality, huge demand but inadequate services seemed to challenge the further development of the mental health service in the PRC. PMID- 16048241 TI - Psychiatric disorders among the Mapuche in Chile. AB - BACKGROUND: The Mapuche are the largest indigenous group in Chile; yet almost all data on the mental health of indigenous populations are from North America. AIM: The study examines the differential DSM-III-R prevalence rates of psychiatric disorders and service utilization among indigenous and non-indigenous community residence. METHODS: The Composite International Diagnostic Interview (CIDI) was administered to a stratified random sample of 75 Mapuche and 434 non-Mapuche residents of the province of Cautin. Lifetime prevalence and 12-month prevalence rates were estimated. RESULTS: Approximately 28.4% of the Mapuche population had a lifetime, and 15.7% a 12-month, prevalent psychiatric disorder compared to 38.0% and 25.7%, respectively, of the non-Mapuche. Few significant differences were noted between the two groups; however, generalized anxiety disorder, simple phobia, and drug dependence were less prevalent among the Mapuche. Service utilization among the Mapuche with mental illness was low. CONCLUSIONS: This is a preliminary study based on a small sample size. Further research on the mental health of indigenous populations of South America is needed. PMID- 16048242 TI - Challenging stigma and discrimination in communities: a focus group study identifying UK mental health service users' main campaign priorities. AB - BACKGROUND: Stigma and discrimination experienced by people with mental health problems have been identified as major obstacles to treatment and recovery. Less is known about how to effectively tackle stigma-discrimination, although there are numerous international, national and local programmes attempting to improve public mental health literacy and anti-discrimination evidenced based practice. AIMS: To explore mental health service users' views on how campaigns to address stigma and discrimination should prioritise their actions. METHOD: Qualitative study using focus group discussions, involving 33 persons aged between 25 and 75. RESULTS: A triad of diminished credibility, dis-empowerment with particular reference to communication problems and avoidance by their social network defined experiences of stigma. Reactions to stigma can be placed in four categories: avoid stigma, resign yourself to it, challenge it, or distance yourself from others with a mental health problem. A range of solutions was discussed with most favouring changes within the health services that are currently supporting them over traditional educational programmes with the public. CONCLUSIONS: For mental health service users stigma must be tackled on many different levels reflecting the varied and complex impact that negative social reactions have on an individual's life. When asked to prioritise one area, most service users in our sample highlighted reforms within the health service for tackling stigma and discrimination. PMID- 16048243 TI - A qualitative study of religious practices by chronic mentally ill and their caregivers in South India. AB - BACKGROUND: Socio-cultural explanatory factors for mental health problems determine help seeking behaviors. The study aimed to understand the reasons mentally ill patients and their families in India choose to seek help from a religious site. MATERIALS AND METHODS: Persons with mental illness and their families were interviewed at religious sites using a guideline questionnaire. Issues such as significant life events, explanations for perceived abnormal behavior and reasons for choosing a specific religious site for 'treatment' were explored. DISCUSSION: Seeking religious help for mental disorders is often a first step in the management of mental disorders as a result of cultural explanations for the illness. This behavior also has social sanctions. PMID- 16048244 TI - Social services day care and health services day care in mental health: do they differ? AB - BACKGROUND: The difference between the services provided by day hospitals and day centres is far from clear. The supposition that day hospitals would provide an acute service, while day centres would offer social support for a more chronic population has been contentious and there is little evidence of how they are currently used. AIMS: We aimed to ascertain the differences between day hospitals (partial hospitalisation) and social service day centres in functions and roles, as perceived by staff, service users and referrers. METHODS: The views of service users and staff at two day hospitals and four day centres were ascertained through questionnaires and interviews, along with those of staff of eight Community Mental Health Teams, who constitute the sole pathway to the two services. RESULTS: Day hospitals were perceived by both referrers and clients to offer short-term, more intensive 'treatment' to more acutely ill people in need of mental health monitoring. Day centres were perceived to offer longer-term support, particularly social support, to people more likely to have longer-term and psychotic illnesses. CONCLUSION: There is currently a clear distinction between day centres and day hospitals, in key features of their services and client groups. It would be unwise to treat them as interchangeable. PMID- 16048245 TI - Community attitudes towards the mentally ill: the results of a national survey of the Taiwanese population. AB - BACKGROUND: In Taiwan, to strengthen the psychiatric rehabilitation system has been one of the primary goals of the Department of Health since 1985. Unfortunately, this endeavor has not been successful and it is believed that one of the barriers is social stigma towards the mentally ill. However, to date no national survey has been conducted for Chinese population on the focal topic using a random sample. AIMS: In this study we explored the attitudes of the general population towards the mentally ill in Taiwan. Specifically, we examined the effect of contact on one's attitudes after controlling for demographic variables. METHODS: A national survey was conducted on 1,203 subjects drawn through a stratified proportional random sampling. Data were collected using the Computer-Assisted Telephone Interview System. We conducted MANOVA and multiple regression analyses to explore the correlates of the attitudes. RESULTS: The results revealed that the general population held relatively higher levels of benevolence, tolerance on rehabilitation in the community, and nonsocial restrictiveness. However, they held relatively less positive attitudes on non authoritarianism and normalization. Overall, direct contact and age were the two most important correlates of community attitudes. Education and occupation were also significant. CONCLUSIONS: Benevolent thoughts do not necessarily guarantee the acceptance of rehabilitation in the community or treating the mentally ill as a person like anyone else. However, the benevolent thought could be transformed into compassion and acceptance of another human being if people are given the opportunity to have direct contact with mentally ill persons. The results also indicated that community education should specifically target laborers of all age groups. PMID- 16048246 TI - Development and validation of the Chinese Version of Indiana Job Satisfaction Scale (CV-IJSS) for people with mental illness. AB - Job satisfaction has been increasingly regarded as an important outcome of vocational rehabilitation programs among people with mental illness. Chinese measures of job satisfaction for individuals with mental illness are however extremely scarce. The aim of this study was to translate the 32-item Indiana Job Satisfaction Scale to Chinese. By means of the expert panel method, the culturally relevant 28-item Chinese Version of Indiana Job Satisfaction Scale (CV IJSS) was then finalized. A validation study among a group of 125 individuals with mental illness showed that the scale had acceptable psychometric properties. Coefficient alpha of the total score was 0.81 with subscales ranging from 0.63 to 0.87. Test re-test reliability as measured by ICC was 0.77 for the total score and ranged from 0.54 to 0.72 for the subscales. Factorial analysis yielded a four factor solution (general satisfaction, job ambiguity and stress, advancement and security, and job recognition) accounting for 44% of the total variance. The factor solution had similarities as well as differences when compared with the Indiana Job Satisfaction Scale. The differences are discussed in the light of cultural differences. Relationship between scores of CV-IJSS and work performance, quality of life and self-esteem was positive in general which may act as evidence to its concurrent validity. The Chinese Version of Indiana Job Satisfaction Scale is ready for use by rehabilitation professionals to assess vocational rehabilitation outcome for individuals with mental illness in Hong Kong and other Chinese societies. Directions for further studies are suggested. PMID- 16048247 TI - Inhibition of nitric oxide synthase reverses permeability changes in a mouse model of acute peritonitis. AB - Acute peritonitis is the most frequent complication of peritoneal dialysis. Previous studies have suggested a major role for nitric oxide (NO) in the permeability changes and loss of ultrafiltration induced by acute peritonitis. In this study, we further investigated the potential role of NO in a mouse model of peritonitis induced by Escherichia coli Lipopolysaccharide (LPS). A 2-hour peritoneal equilibration test was performed in control and LPS-treated mice using 7% glucose dialysate supplemented or not with the NO synthase inhibitor NG-nitro L-arginine methyl ester (L-NAME). The levels of NO metabolites in the dialysate were maximal 18 hours after LPS injection. At that time, acute peritonitis induced by LPS was reflected by an increased recruitment of leukocytes, an increased intraperitoneal release of interleukin-6, a significant increase in the peritoneal permeability for small solutes, a loss of sodium sieving, and a loss of ultrafiltration in comparison with controls. Addition of L-NAME in LPS-treated mice significantly reversed permeability modifications and prevented the release of NO metabolites into the dialysate. These results confirm that increased NO mediates permeability modifications during acute peritonitis, and illustrate the potential of mouse models to investigate the molecular mechanisms regulating peritoneal permeability. PMID- 16048248 TI - Transforming growth factor-beta: importance in long-term peritoneal membrane changes. AB - We have provided evidence from adenovirus-mediated gene transfer to the peritoneum that transforming growth factor-beta1 mimics many of the functional and structural changes in the peritoneum in patients on long-term peritoneal dialysis, including fibrosis, increased submesothelial thickness, angiogenesis, increased solute transport, and ultrafiltration dysfunction. We review several key properties of this important fibrogenic molecule. PMID- 16048249 TI - Effects of inhibition of the polyol pathway during chronic peritoneal exposure to a dialysis solution. AB - BACKGROUND: Peritoneal dialysis with glucose- and lactate-containing dialysis solutions stimulates peritoneal angiogenesis and fibrosis. These serious side effects can also be induced by chronic peritoneal exposure to dialysis solutions in nonuremic rats. The high glucose concentrations of the dialysis solutions may saturate physiological glucose metabolism pathways and stimulate the polyol pathway that has been described to damage nerves and vessels in diabetes mellitus. To investigate the role of the polyol pathway in the development of fibrosis and angiogenesis during chronic peritoneal exposure, the rate-limiting aldose reductase activity in the polyol pathway was inhibited in a chronic peritoneal exposure model in the rat, in which different administration routes were compared. EXPERIMENTAL PROCEDURES: Three groups of rats received daily intraperitoneal infusion with lactate/glucose (3.86%)--containing dialysate via a peritoneal catheter with a subcutaneous puncture device, for 14 weeks: group 1 received only the dialysis solution, groups 2 and 3 received, in addition, zopolrestat, administered either orally (group 2) or intraperitoneally (group 3). After sacrifice, omental tissue was examined by histology for the presence of fibrosis (Picro Sirius Red) and the number of blood vessels (CD31). RESULTS: Histology revealed significantly less Picro Sirius Red-positive tissue in perivascular areas of both experimental groups and submesothelial areas of the oral group in comparison to the control group. There were significantly fewer CD31-positive vessels perfield in both groups treated with zopolrestat compared to the infusion-only group: group 2, 9 (7 - 12); group 3, 17 (13 - 38), compared to group 1, 37 (32 - 39), p < 0.05. CONCLUSION: The combination of peritoneal exposure to dialysis fluids and administration of zopolrestat, a newly developed inhibitor of aldose reductase activity, resulted in less fibrosis and fewer peritoneal vessels than exposure to dialysis fluids only, in a long-term exposure model in the rat. Inhibition of the polyol pathway may thus offer an important contribution to allow long-term continuation of peritoneal dialysis. PMID- 16048250 TI - The effect of low glucose degradation product dialysis solution on epithelial-to mesenchymal transition in continuous ambulatory peritoneal dialysis patients. AB - BACKGROUND: Human peritoneal mesothelial cells (HPMCs) undergo a transition from an epithelial phenotype to a mesenchymal phenotype (EMT) during peritoneal dialysis (PD). That transition may be directly related to failure of peritoneal membrane function. OBJECTIVE: In a randomized prospective controlled study, we investigated the effect of low glucose degradation product (GDP) dialysis solution on the transition of HPMCs. METHODS: Among new continuous ambulatory PD patients, 60 patients completed a 12-month protocol (low-GDP solution group, n = 32; high-GDP solution group, n = 28). At the 1st, 6th, and 12th months, HPMCs drained from overnight effluent were cultured on T25. When they had nearly reached confluence, cell scores were measured blindly by the same person (score 1 = cobblestone-shaped HPMCs, score 2 = mixed, score 3 = fibroblast dominant). Cell scores and clinical indices, including peritoneal markers, were compared between the low-GDP and high-GDP groups at the 1st, 6th, and 12th months. The factors associated with EMT were analyzed with generalized estimating equations using STATA 7.0 (STATA Corp., College Station, Texas, USA). In addition, vimentin and cytokeratin 8/18 stains were used to verify EMT in cultured cells and peritoneal specimens in some patients. RESULTS: (1) The low-GOP group showed higher dialysate cancer antigen 125 levels from the 1st to 12th months (55.4 +/- 24.8 vs 8.8 +/- 1.7, 56.7 +/- 28.1 vs 22.1 +/- 11.3, and 54.2 +/- 28.2 vs 24.6 +/- 16.5 U/mL, at the 1st, 6th, and 12th months, respectively; all p = 0.000). (2) The low GOP group showed lower cell scores at the 1st, 6th, and 12th months (1.22, 1.22, and 1.56 vs 1.61, 1.75, and 2.14; p < 0.05, p < 0.01, and p < 0.01, respectively). (3) At the 12th month, the number of fibroblast-dominant cultures (score 3) was significantly lower in the low-GOP group [4/32 (12.5%) patients vs 14/28 (50%), p < 0.05]. (4) Both cobblestone-shaped HPMCs and fibroblastoid cells were positively stained with cytokeratin and vimentin. (5) There were many cytokeratin- and vimentin-positive cells in the submesothelial area in the peritoneal biopsy specimens. (6) The consistent factor associated with EMT was only high-GOP solution (60 patients, n = 178, beta coefficient 0.312, p = 0.000; 46 patients, n = 137, beta coef: 0.228, p = 0.000) and not numbers of peritonitis episodes, duration of angiotensin-converting enzyme inhibitor and/or angiotensin receptor blocker medication, or diabetes. CONCLUSION: Low-GDP solution showed beneficial effects such as rapid remesothelialization and less EMT in the peritoneum with time on PD. PMID- 16048251 TI - Osmotic agents hamper mesothelial repopulation as seen in the doughnut in vivo model. AB - BACKGROUND: The problem of mesothelial cell injury derived from the use of peritoneal dialysis solutions has been explored deeply. Conversely, the eventual detrimental effects upon mesothelial cell regeneration have awaked less investigative efforts than those focused on injury. OBJECTIVE: To evaluate in the in vivo and in situ rat "doughnut" model of mesothelial repopulation, the eventual effect of peritoneal lavage with Hank's Balanced Salt Solution (HBSS) as well as that of 4.25% glucose and 7.5% icodextrin dialysis solutions. EXPERIMENTAL ANIMALS: 100 Sprague-Dawley albino rats were included in the study. Animals were divided into five groups of 20 rats each: group 1: control at zero time; group 2: sham-injected rats; group 3: rats exposed to HBSS; group 4: rats treated with 4.25% glucose peritoneal dialysis solution; group 5: rats injected with 7.5% icodextrin. METHODS: Selective exfoliation of a ring of mesothelium (width 0.8 mm, diameter 4 mm) covering the anterior surface of the liver was performed in 80 animals. The control zero-time group was used to evaluate the normal density distribution of the mesothelial cells forming the monolayer. The other groups were treated by means of daily sham injections or intraperitoneal infusion of each experimental solution for a period of 30 consecutive days. After a recovery period of 15 days, imprints and biopsies from the monolayer covering the exfoliated area were taken and processed for light microscopy. RESULTS: Macroscopic observation of the abdominal cavity at the end of the 15-day recovery period showed that the prevalence of fibrotic adhesions between the peritoneal exfoliated area and the neighboring diaphragm was 10% forthe sham-injected group, 5% for the HBSS-exposed animals, 85% for the rats injected with 4.25% glucose, and 95% for the icodextrin-treated group. Prevalence of fibrous adhesions in sham injected animals and rats exposed to HBSS were devoid of statistically significant differences. Conversely, comparison of these groups with results observed in animals treated with the osmotic agents was significant, at the p < 0.0039 level. Regarding density distribution of mesothelial cells observed in imprints, there were no significant differences between the control zero-time and the sham-injected group. This parameter was marginally lower (p < 0.05) in the HBSS-treated rats. Imprints were not taken from animals exposed to glucose or icodextrin because a dense layer of connective tissue replaced the exfoliated mesothelial area. CONCLUSIONS: Observations made in this study support the contention that both osmotic agents, 4.25% glucose and 7.5% icodextrin, substantially restrain the normal process of mesothelial cell repopulation and induce repair by means of connective tissue. The underlying mechanism is most likely sustained oxidative stress. PMID- 16048252 TI - Effects of peritoneal dialysis solutions on the peritoneal membrane: clinical consequences. AB - This review provides an overview of recent studies that show the clinical significance of biocompatibility of peritoneal dialysis fluids. PMID- 16048253 TI - Lessons from basic research for PD treatment. AB - Over the past 30 years, the focus of peritoneal dialysis research has changed from the technical issues related to the establishment of clinical peritoneal dialysis to complex problems of peritoneal membrane biology. Here, we present how these research topics developed, discuss their significance for clinical science, and outline future challenges for peritoneal dialysis research. PMID- 16048254 TI - The difference in causes of early and late ultrafiltration failure in peritoneal dialysis. AB - OBJECTIVE: Ultrafiltration failure (UFF) is a major complication of peritoneal dialysis. Although it seems associated with long-term treatment, it can also occur in recently started patients. To identify the causes of this complication in patients with early and late UFF we studied a group of 48 patients. Patients were classified as early if they had been treated for less than 2 years and as late if they had been treated for more than 4 years. METHOD: The patients were studied using a standard peritoneal permeability analysis. They all had a net ultrafiltration of less than 400 mL after a 4-hour dwell with 3.86% glucose. As possible causes for UFF, the solute transport parameters dialysate-to-plasma ratio (D/P) and mass transfer area coefficient of creatinine were compared, as well as the effective lymphatic absorption rate (ELAR) and the maximum dip in D/P sodium as an assessment of osmotic conductance to glucose. RESULTS: 25 short-term patients were compared with 23 long-term patients. Both groups showed an equal distribution of high small solute transport rates as a cause of UFF. The chi square test showed that a high ELAR was a more frequent cause in early UFF compared to late UFF. However, a decreased osmotic conductance to glucose was significantly more often observed in late UFF. Some patients showed more than one cause of the complication. CONCLUSION: This study has shown that UFF in long-term patients is often caused by a decreased osmotic conductance to glucose, most likely caused by a dysfunction of peritoneal water channels in combination with increased peritoneal surface area. In short-term patients, aquaporin dysfunction is rare, but a high ELAR was a very important factor in the occurrence of UFF. PMID- 16048255 TI - A ward-based procedure for assessment of fluid status in peritoneal dialysis patients using bioimpedance spectroscopy. AB - OBJECTIVE: This study was carried out to evaluate a procedure for converting bioimpedance spectroscopy (BIS) data to a clinically useful measure of fluid status that is easily interpreted by nursing staff and patients. DESIGN: The ratio of extracellular water (ECW) to total body water (TBW) in a cohort of peritoneal dialysis (PD) patients was compared with the distribution for normal control subjects. The result was expressed as the difference between the measured ECW/TBW ratio and the mean ratio for age- and sex-matched controls divided by the standard deviation for the controls (the "hydration score"). Where possible, interventions were made to reduce the target weight in patients with a hydration score greater than +2.0. SETTING: This clinical study was carried out in the nurse-led outpatient PD unit at St. James's University Hospital. BIS measurements were carried out during routine clinic visits. The data for the control subjects were obtained in a range of settings in the UK and the Czech Republic. PATIENTS: 31 PD patients (21 M, 10 F; aged 19 - 78 years) who came to the PD unit for routine procedures during the study participated. RESULTS: The mean hydration score for PD patients was significantly higher than for the control subjects (+1.3 vs 0.0, p < 0.0001). 11 (35%) patients had a hydration score greater than +2.0, compared with only 2.5% of the controls; systolic blood pressure was greater than 130 mmHg in only two of these patients. After a 3-month follow-up, a weight reduction of 3.6 +/- 2.3 kg had been achieved in 7 patients through modality change and implementation of prescription changes, with no adverse effects. CONCLUSIONS: Although this was a small study, all the data collected indicated that the hydration score provides a reliable indication of fluid status in PD patients. BIS measurements are now carried out at 6-month intervals and are used to monitor the effect of interventions. PMID- 16048256 TI - The effect of normalization of ECW volume as a marker of hydration in peritoneal dialysis patients and controls. AB - OBJECTIVES: We measured extraceLlular water (ECW) and intracellular water (ICW) volumes in peritoneal dialysis (PD) patients and controls to determine the effect of ICW variation on ECW/ICW ratio and to compare alternative ratios of ECW to height, height2, weight, and body surface area (BSA). PATIENTS AND METHODS: We measured body water compartments by deuterium oxide and bromide dilution in 29 PD patients (14 M, 15 F) and 31 controls (15 M, 16 F). RESULTS: ECW was similar in PD patients (17.58 +/- 3.58 L) and controls (17.20 +/- 2.97 L), p = NS. ICW was nonsignificantly lowerin PD patients (17.58 +/- 4.88 L) than in controls (19.71 +/- 5.08 L), p = NS. ECW/ICW was greaterin PD patients (1.06 +/- 0.32) than in controls (0.92 +/- 0.25), p = 0.057, and was inversely correlated with ICW in PD patients (r = -0.733, p < 0.0001) and controls (r = -0.721, p < 0.0001). In contrast, ECW/height, ECW/height2, ECW/weight, and ECW/BSA were similar for the two groups. CONCLUSIONS: ECW/ICW is affected by changes in ICW as well as by ECW varying with hydration. ECW/ICW ratio leads to the spurious impression of overhydration in subjects with smaller ICW volumes. ECW/ICW does not reflect hydration alone and other methods of expressing ECW as a measure of hydration need further evaluation. PMID- 16048257 TI - Association of vitamin D receptor gene polymorphisms with hypercalcemia in peritoneal dialysis patients. AB - BACKGROUND: Some polymorphisms at the human vitamin D receptor (VDR) gene locus may influence calcium and bone metabolism. We investigated the roles of the BsmI and TaqI VDR gene polymorphisms in the development of hypercalcemia in Turkish peritoneal dialysis (PD) patients. METHODS: We enrolled 132 PD patients treated with dialysate containing 1.75 mmol/L calcium. Serum levels of calcium, phosphorus, albumin, and intact parathyroid hormone (iPTH), and the cumulative doses of calcium-based phosphate binders and calcitriol were recorded every 3 months. The VDR BsmI and TaqI genotypes were determined by polymerase chain reaction. RESULTS: When the patients were categorized according to these VDR genotypes, serum levels of phosphorus and iPTH and cumulative doses of calcium based phosphate binders and calcitriol were similar across groups. The corrected serum calcium levels tended to increase in the patients with BsmI non-BB (Bb + bb) variants, but were significantly decreased in the BB variants (9.9 +/- 0.7 vs 9.1 +/- 0.6 mg/dL, p < 0.05). Hypercalcemia appeared in 21.2% of the patients during the follow-up period. The hypercalcemic patients had a significantly higher prevalence of the BsmI non-BB genotype than the normocalcemic patients (85.7% vs 59.6%, p < 0.007). On the contrary, the serum calcium levels were not affected by the TaqI VDR gene polymorphism (p > 0.05). CONCLUSIONS: These findings suggest that the non-BB variants of the BsmI VDR gene polymorphism are associated with increased risk of developing hypercalcemia in PD patients. PMID- 16048258 TI - Analysis of patient flow into dialysis: role of education in choice of dialysis modality. AB - BACKGROUND: Despite advances in predialysis care, morbidity and mortality remain high. OBJECTIVES: To analyze end-stage renal disease (ESRD) patient demographics and clinical data on education on dialysis treatment options, type of chronic renal replacement therapy (RRT), and effects of planned versus non-planned dialysis start. METHODS: 621 patients, from 24 Spanish hospitals, who started RRT in 2002. Peritoneal or vascular access at dialysis initiation was considered "planned." RESULTS: 304 (49%) patients were non-planned and half of them had prior nephrology follow-up. Of the patients with >3 months nephrology follow-up (76% of all), only half were educated on dialysis modalities. Dialysis education was associated with planned start in 73.4% versus 26% in non-educated patients (p < 0.05), shorter follow-up (55 vs 65 months, p = 0.033), more medical visits in the prior year (6.5 vs 4.4, *p < 0.001), more patients starting peritoneal dialysis (31% vs 8.3%*), and more specific follow-up by ESRD unit versus general nephrology care (63% vs 26%*). Non-planned start was associated with older age (63 vs 60.6 years, p = 0.06), fewer medical visits (4.6 vs 6.4*), less education about modality options, and greater use of hemodialysis (92% vs 75%*). Planned patients had better biochemical parameters at start of dialysis. CONCLUSION: Despite nephrology follow-up, half the patients did not have a planned dialysis start. Planned start was associated with better clinical status. More patients chose peritoneal dialysis when educated about dialysis modality options. ESRD specific units were more likely to provide patient education. PMID- 16048259 TI - Assisted automated peritoneal dialysis (AAPD) for the functionally dependent and elderly patient. AB - OBJECTIVE: To describe basic demographics and clinical outcomes among elderly end stage renal disease (ESRD) patients physically dependent on a caregiver and maintained on an assisted automated peritoneal dialysis (AAPD) program. DESIGN: Retrospective single-center study based on patient records and data files. SETTING: University Hospital. PATIENTS: 64 physically dependent AAPD patients followed for 1.012 treatment months. Assistance and care was delivered by 52 briefly trained teams of visiting nurses or nursing home staff. RESULT: Crude 1 year survival was 58% and 2-year survival was 48%. Crude 1- and 2-year survivals, excluding deaths within 90 days, were 66% and 54% respectively. We found no significant effect on survival by main causes of ESRD, gender, age, late referral, need for acute start, social isolation, physical dependency on help at inclusion, or residence in a nursing home. 10% of patient-days on AAPD were spent in hospital. 13 (20%) of the patients were converted permanently to hemodialysis due to PD technique failure. The incidence of peritonitis was 1 in every 25.3 treatment-months. CONCLUSIONS: AAPD may be a feasible and safe option for renal replacement therapy for frail, elderly, and physically dependent patients with ESRD. Despite the special patient selection for this AAPD program, we achieved results of international standards for patient survival, PD technique survival, and incidence of acute peritonitis. These results do notjustify withholding dialysis from this group of patients. PMID- 16048260 TI - Initiating CAPD with a regimen low in glucose and glucose degradation products, with icodextrin and amino acids (NEPP) is safe and efficacious. AB - BACKGROUND: The high Levels of glucose, glucose degradation products (GDPs), and lactate buffer present in standard peritoneal dialysis (PD) solutions contribute to peritoneal damage, malnutrition, and dyslipidemia. Therefore, we studied the feasibility of a PD regimen as low as possible in glucose and GDPs. METHODS: In a prospective 30-week study, patients new to continuous ambulatory PD (CAPD) were randomized to either a standard PD regimen (SPD; 4 dwells glucose-/lactate-based) or a low glucose-GDP regimen (NEPP; 1 dwell amino acids, 1 dwell icodextrin, and two dwells bicarbonate/lactate-buffered glucose-based solution). RESULTS: Results obtained during a 30-week study period for 63 new CAPD patients (30 NEPP, 33 SPD) were analyzed. Intraperitoneal glucose load was lower in the NEPP group (111 +/- 76 vs 159 +/- 40 g/day at 30 weeks, p < 0.001). Dialysis efficacy, ultrafiltration, weight, blood pressure, and laboratory results were similar in the groups, whereas, in the NEPP group, cancer antigen 125 in dialysate effluents decreased less but dialysate-to-plasma ratios were slightly higher. CONCLUSION: Short-term treatment of new CAPD patients with a PD regimen low in glucose and GDPs is feasible. Dialysis efficacy, ultrafiltration, and metabolic consequences are similar to those during a standard glucose-lactate-based regimen, whereas peritoneal transport seems slightly higher and preservation of mesothelial cell mass better during NEPP. PMID- 16048261 TI - When to start dialysis treatment: where do we stand? AB - BACKGROUND: Since the publication of opinion-based guidelines regarding the timing of dialysis treatment, there has been a trend toward earlier initiation. OBJECTIVE: In this review, the existing guidelines and the currently published studies that evaluate them are discussed. RESULTS: These studies could not demonstrate a clear benefit on survival or quality of life for patients who started with relatively higher renal function. CONCLUSION: Early start of dialysis treatment should not be confused with early referral to the nephrologist. It is concluded that initiation of dialysis should not depend on a predefined magnitude of renal function, but should be tailored to the individual patient. PMID- 16048262 TI - Improving salt balance in peritoneal dialysis patients. AB - Volume homeostasis is an important predictor of outcome in peritoneal dialysis. Because volume retention is driven by salt retention, maintenance of salt balance should be a concern to all nephrologists. An important factor in this is dietary salt restriction. This has long been neglected in peritoneal dialysis patients, where it was considered that the continuous nature of the therapy and residual renal function would be sufficient to remove any extra salt load. In patients with preserved renal function, diuresis and salt excretion can be enhanced by the application of high doses of loop diuretics. This practice seems not to have an impact on the further deterioration of renal function. Peritoneal salt removal can be enhanced using polyglucose. Also, the use of low sodium-containing dialysate can be efficient. These solutions are, however, not commercially available, and they need higher concentrations of glucose to obtain an efficient osmolarity. It should always be considered that, due to sodium sieving over the ultrasmall pores, fluid and salt removal are not always concordant. PMID- 16048263 TI - Peritoneal dialysis prescription for diabetic patients. AB - The number of end-stage renal disease (ESRD) patients with diabetes mellitus has increased dramatically during the past few years and, in many countries, diabetes has become the most important cause of ESRD in patients admitted to dialysis. Furthermore, compared to nondiabetic patients, diabetic patients continue to suffer from more frequent and severe comorbidity and complications, including cardiovascular disease, poor fluid balance, worse quality of life, as well as high morbidity and mortality after initiation of dialysis. These systemic problems in diabetic patients should influence the dialysis prescription. In addition, the structure and transport properties of the peritoneal membrane may deteriorate as a consequence of diabetes. Thus, both the systemic and the peritoneal consequences of diabetes influence the dialysis prescription in diabetic patients. In this brief review, we discuss the care of diabetic ESRD patients on peritoneal dialysis--which, compared with hemodialysis, has both advantages and disadvantages in this group of patients--focusing on the special needs for intense and integrated care involving individualized dialysis prescription as well as care of diabetic complications and comorbidity in this diseased patient group. PMID- 16048264 TI - The clinical and epidemiological aspects of vascular mortality in chronic peritoneal dialysis patients. AB - This article reviews the clinical aspects of and epidemiological links between vascular mortality and the dialysis population, and emphasizes areas that warrant further clarification. In particular, we highlight potential pitfalls in interpretation of published observational and clinical studies, notably some of the issues related to reverse epidemiology of risk factors for cardiovascular disease. Recent published data from our own center in the Prince of Wales Hospital relating to the significance of residual renal function, inflammation, valvular calcification, as well as left ventricular hypertrophy were highlighted. Actions are needed to tackle both the traditional and the nontraditional factors for cardiovascular disease in order to treat this problem causing the highest mortality in peritoneal dialysis patients. PMID- 16048265 TI - Clinical epidemiology of major nontraditional risk factors in peritoneal dialysis patients. AB - BACKGROUND: End-stage renal disease (ESRD) is a situation with a cardiovascular (CV) risk profile of almost unique severity. While traditional risk factors dominate the scene in the general population, in chronic kidney disease (CKD), nontraditional risk factors play an increasingly important role, being perhaps dominant in ESRD patients. OBJECTIVE: We review the role inflammation [C-reactive protein (CRP)], hyperhomocysteinemia, high plasma norepinephrine, and accumulation of the endogenous inhibitor of the nitric oxide synthase asymmetric dimethylarginine (ADMA) in the high all-cause and CV mortality of patients on continuous ambulatory peritoneal dialysis (CAPD). RESULTS: The association between CRP and clinical outcomes in patients on peritoneal dialysis (PD) was examined in six studies totaling 692 subjects. The largest of these studies in Caucasians indicates that the independent risk of CV events in patients in the top CRP quartile is about five times higher than in the bottom quartile. Seven prospective studies, including over 1000 hemodialysis and 176 CAPD patients, reported both positive and negative associations between homocysteine and mortality and/or CV events. Because homocysteine circulates bound to albumin, negative associations--rather than negating the vasculotoxicity of homocysteine- most likely reflect the very deleterious effects of malnutrition. Plasma norepinephrine is higher in CAPD than in hemodialysis patients, and multivariate analyses suggest the difference quantitatively entails a 16% higher risk of incident CV events. Likewise, ADMA is more elevated in CAPD patients and such an elevation corresponds to a 15% increase in risk. CONCLUSION: Nontraditional risk factors are far more prevalent in ESRD patients than in the general population. ADMA and norepinephrine may play a greater role in CV risk in CAPD than in hemodialysis patients. PMID- 16048266 TI - Peritoneal dialysis in elderly patients: clinical experience. AB - Many older patients do not cope well with hemodialysis, yet the proportion of older patients on peritoneal dialysis (PD) is considerably lower than for younger patients. This is despite the fact that clinical outcome studies show that older patients cope as well as, if not better than, younger patients on PD. Furthermore, the North Thames Dialysis Study, a prospective study of patients > or = 70 years old, has shown that survival, hospitalization, and quality of life were identical for patients on hemodialysis and patients on PD. PMID- 16048267 TI - Individualized PD prescription: APD versus CAPD. AB - The proportion of patients performing automated peritoneal dialysis (APD) is increasing worldwide, a development probably caused by the better possibilities of adapting APD to the patients' individual needs with respect to private life as well as dialysis adequacy. Patients prefer the independence from dialysis during the day and report a higher quality of life compared to patients on continuous ambulatory peritoneal dialysis (CAPD). In case of declining clearance rates, Kt/V, or sodium removal rates, a change in the APD regimen, together with higher fill volumes and, for example, combination with daytime CAPD, offers the tools to increase the dialysis dose as required by the individual clinical situation. The development of an online dialysis solution production system for APD could even improve the possibilities of individualizing peritoneal dialysis by providing variable concentrations of glucose, sodium, and bicarbonate buffer. PMID- 16048268 TI - Impact of peritoneal dialysis dose guidelines on clinical outcomes. AB - The objective was to review the rationale for the Kidney Disease Outcomes Quality Initiative (K/DOQI) recommendations for adequacy of peritoneal dialysis and to evaluate the impact of these recommendations on clinical practice and patient survival. The K/DOQI recommendations were based on large observational studies; the target weekly Kt/V value of 2.0 assumed equivalence of peritoneal and renal clearances. This assumption is no longer considered correct. The impact on clinical practice was evaluated by an examination of temporal trends before and after publication of the guidelines in 1997. In the United States and The Netherlands, there had been a trend toward increased delivered total Kt/V prior to 1997, and there was no acceleration in this trend after 1997. Two randomized clinical trials have implemented these guidelines with increased peritoneal Kt/V (or creatinine clearance) used to achieve the K/DOQI target in the intervention group. This was not associated with improved survival, compared to a lower Kt/V, in either of the randomized clinical trials. Among the explanations for the failure to improve outcome are potential adverse effects of increasing the dialysis dose. These include increased intraperitoneal pressure associated with increased exchange volume, failure to increase clearance of middle molecules, and increased exposure to glucose. Strategies that increase peritoneal clearance without exposure to these potential adverse effects include more-frequent exchanges rather than increased exchange volume, and decreased exposure to glucose and glucose degradation products. Pending such studies, current K/DOQI guidelines should be updated in a timely manner. PMID- 16048269 TI - The impact of guidelines for the prevention of anemia on clinical outcome. AB - Guidelines for anemia management in renal disease are supported by substantial evidence demonstrating improvement in quality of life and objective markers of physical and cognitive performance. Randomized control studies demonstrating a survival benefit or improved cardiovascular outcomes are inconsistent. However, observational studies clearly demonstrate reduced mortality and hospitalization rate in patient cohorts on hemodiaLysis with hemoglobin measurements within the recommended target ranges. Data from patients in the predialysis phase of chronic kidney disease and those on peritoneal dialysis are limited and studies assessing the clinical impact of adherence to guidelines should be further explored in these populations. Available evidence suggests a proactive approach to anemia management should be practiced. PMID- 16048270 TI - Access and continuous flow peritoneal dialysis. AB - Continuous flow peritoneal dialysis (CFPD) presents many potential advantages over any conventional peritoneal dialysis modality. Foremost are the impressive clearances achieved by CFPD in clinical studies, approaching those of quotidian hemodialysis. The principal limitation of CFPD is its inability to provide adequate and consistent ultrafiltration in clinical practice, contrary to the high ultrafiltration predicted by theoretical models. There is a good possibility that this discrepancy is due to poor mixing, streaming, and recirculation of dialysate related to inadequate peritoneal access. Novel access designs are encouraging but await longitudinal clinical testing to prove their superiority. PMID- 16048271 TI - Encapsulating peritoneal sclerosis is a separate entity: Con. AB - There is controversy with respect to the issue that encapsulating peritoneal sclerosis (EPS) is a separate entity from simple sclerosis (SS), which is the uniform change of the peritoneum on peritoneal dialysis. These following ideas support the notion that the development of EPS is connected with pathology that is not the same as that responsible for SS: (1) EPS is a rare disorder, (2) certain factors are involved with the etiology, (3) the clinical background of the patients is not uniform, and (4) the histopathological findings of EPS are different from those of SS. There are pitfalls in these concepts and they need be revised. This paper discusses the issue against the idea that EPS is a separate entity, supporting the idea that it has a strong connection with SS in its primary pathophysiology. PMID- 16048272 TI - Sclerosing peritonitis: a nosological entity. AB - The peritoneal histology of 224 peritoneal dialysis (PD) patients without sclerosing peritonitis (SP) and of 39 PD patients with SP was evaluated. Of the 224 patients, 180 showed simple sclerosis (SS). In these subjects, slight thickness of sclerosis (10 - 70 microm), slight parvicellular infiltration (5/180), slight arterial thickening with no vessel occlusion (19/180), and slight tissue calcification (1/180) were observed. In the 39 patients with SP, striking histological changes versus SS were detected: thickness of sclerosis 250 - 4000 microm, p < 0.01; inflammation 39/39, p < 0.01 (parvicellular infiltration 36/39, p < 0.01; microabscesses 15/39, p < 0.05; giant cells 38/39, p < 0.01; granulation tissue 38/39, p < 0.01); arterial alterations 39/39, p < 0.01 (thickening 39/39, p < 0.01; occlusion 39/39, p < 0.01; calcification 26/39, p < 0.01; ossification 9/39, p < 0.01); tissue calcification 12/39, p < 0.01 (with ossification 4/39, with bone marrow 2/39). The thickness of sclerosis in SS was higher in parietal (30 - 70 microm) than in visceral peritoneum (10 - 40 microm, p < 0.05); in SP it was higher in visceral (600 - 4000 microm) than in parietal peritoneum (250 - 2000 microm, p < 0.05). These striking differences suggest consideration of SS and SP as two separate nosological entities. Differences in frequency, animal models, etiology, and clinical impact seem to confirm this hypothesis, showing that SP is not just the evolution of SS. PMID- 16048273 TI - The development of encapsulating peritoneal sclerosis. PMID- 16048274 TI - Prevention and treatment of peritoneal dialysis-associated peritonitis in pediatric patients. AB - Peritoneal dialysis (PD) is the preferred dialysis modality in children and adolescents aged less than 15 years. Peritoneal dialysis-associated peritonitis remains a major cause of morbidity and reason for dropout from the PD program, although the incidence of peritonitis seems to have decreased during the past few years. Improved patient care, more frequent use of automated peritoneal dialysis (APD), use of PD catheters with downward facing exit sites, and Staphylococcus aureus prophylaxis account for this decline in infectious complications. With respect to the isolated micro-organism in PD-associated peritonitis, a predominance of gram-positive germs is found in children. Recent registry data suggest a decrease in coagulase-negative staphylococci, with a relative increase in gram-negative peritonitis episodes. The empiric antibiotic treatment regimen using a first-generation cephalosporin or a glycopeptide in combination with a third-generation cephalosporin in a risk-stratified manner was suggested in the pediatric peritonitis treatment guidelines. This regimen is currently being evaluated in the International Pediatric Peritonitis Registry. PMID- 16048275 TI - Cardiovascular morbidity and mortality in children with chronic kidney disease in North America: lessons from the USRDS and NAPRTCS databases. AB - Cardiovascular (CV) disease mortality has not changed in the past three decades, remaining the second most common cause of death in children with end-stage renal disease (ESRD). This article reviews pediatric data pertaining to CV disease obtained from the two largest renal databases in North America. The data indicate high incidence and prevalence of CV complications and cardiac risk factors in children with ESRD, accounting for high cardiac mortality in this patient population. Early identification and intervention to treat modifiable risk factors and asymptomatic cardiac disease might lead to prevention of cardiac disease and to a decrease in CV morbidity and mortality in young adults who developed chronic kidney disease during childhood. PMID- 16048276 TI - Cardiovascular disease as a late complication of end-stage renal disease in children. AB - OBJECTIVE: To analyze the late cardiovascular outcome of end-stage renal disease (ESRD) in children. DESIGN: A nation-wide long-term follow-up study. Determinants of outcomes and causes of death were retrospectively assessed. Patients underwent assessment of overall health state, B- and M-mode ultrasound of the carotid arteries, and echocardiography for cross-sectional analysis. RESULTS: We analyzed the medical course of all 249 adult Dutch patients with ESRD onset between 1972 and 1992 at age 0 - 14 years, and who were born before 1979. Of the 187 living patients, 140 participated in the cross-sectional part of the study. The standardized mortality rate was 31.0. Overall 5-, 10-, and 20-year survival after ESRD onset was 87%, 82%, and 78%, respectively. Cardiovascular disease accounted for most deaths (41%). In the whole group, left ventricular hypertrophy (LVH), aortic valve calcification, and arterial wall stiffening were highly prevalent. LVH was associated with hypertension at time of assessment. Aortic valve calcification was strongly associated with a long total duration of peritoneal dialysis (beta = 0.33, p < 0.001). Arterial wall pathology was not associated with current treatment modality. CONCLUSIONS: As in adults, cardiovascular disease is the most important cause of death in children with ESRD. Stricter reduction of volume overload, prevention of high serum calcium-phosphate product, and more vigorous treatment of hypertension are important targets to improve cardiovascular survival in children with ESRD. PMID- 16048277 TI - Endothelial function in pediatric patients on peritoneal dialysis: the need for data. AB - Cardiovascular complications are emerging as the primary cause of death for patients with childhood end-stage renal disease. Children with end-stage renal failure are subjected to many of the risk factors for cardiovascular disease identified in adult patients. Dysfunction of the endothelium is presently regarded as a first but reversible step in the development of atherosclerosis. Noninvasive techniques to assess endothelial function have been recently developed and have been proven to predict future mortality in adult patients. These techniques are readily applicable to pediatric patients. Endothelial dysfunction has been demonstrated in children in all stages of renal failure. Data on pediatric patients treated with peritoneal dialysis are currently lacking, however. Considering the abundance of cardiovascular risk factors specific to treatment with peritoneal dialysis, such studies should be initiated. PMID- 16048278 TI - The international pediatric peritonitis registry: a global Internet-based initiative in pediatric dialysis. AB - OBJECTIVES: Recommendations for the management of peritoneal dialysis-associated peritonitis in children have recently been developed by an International Society for Peritoneal Dialysis (ISPD) expert committee. The International Pediatric Peritonitis Registry (IPPR) was established in October 2001 as a global consortium of 47 pediatric dialysis centers in order to assess the validity of these guidelines. DESIGN: The IPPR is an internet-based registry collecting data on pediatric peritonitis episodes treated according to the ISPD guidelines. Data on 375 episodes have been collected as of July 2004. DATA ACQUISITION: Detailed data are obtained online on the diagnosis of peritonitis, antibiotic and adjunctive therapy, as well as on possible risk factors and treatment results. CONCLUSIONS: Final data analysis of the IPPR will yield extensive information on the treatment and outcome of peritoneal dialysis-associated peritonitis in children. PMID- 16048279 TI - How to increase adequacy of peritoneal dialysis in children? AB - Since children on dialysis are treated most often with nightlyintermittent peritoneal dialysis, adequacy of dialysis is determined by the number and duration of cycles, the volume of the dialysis fluid applied, and the choice of dialysis solution. The number and duration of cycles are dependent on the maximal acceptable duration of night rest and the permeability properties of the peritoneal membrane. The latter can be established by performance of a peritoneal equilibration test. The volume used should be about 1200 mL/m2 body surface area, and intraperitoneal pressure should be between 5 and 15 cm H2O. The dialysis solution administered should have a glucose concentration as low as possible, and an icodextrin daytime dwell may be considered. PMID- 16048280 TI - The influence of peritoneal surface area on dialysis adequacy. AB - In children, the prescription of peritoneal dialysis is based mainly on the choice of the peritoneal dialysis fluid, the intraperitoneal fill volume (mL/m2 body surface area (BSA)], and the contact time. The working mode of the peritoneal membrane as a dialysis membrane is more related to a dynamic complex structure than to a static hemodialyzer. Thus, the peritoneal surface area impacts on dialysis adequacy. In fact, the peritoneal surface area may be viewed as composed of three exchange entities: the anatomic area, the contact area, and the vascular area. First, in infants, the anatomic area appears to be two-fold larger than in adults when expressed per kilogram body weight. On the other hand, the anatomic area becomes independent of age when expressed per square meter BSA. Therefore, scaling of the intraperitoneal fill volume by BSA (m2) is necessary to prevent a too low ratio of fill volume to exchange area, which would result in a functional "hyperpermeable" peritoneal exchange. Second, the contact area, also called the wetted membrane, is only a portion of the anatomic area, representing 30% to 60% of this area in humans, as measured by computed tomography. Both posture and fill volume may affect the extent of recruitment of contact area. Finally, the vascular area is influenced by the availability of both the anatomic area and the recruited contact area. This surface is governed essentially by both peritonealvascular perfusion, represented by the mesenteric vascular flow and, hence, by the number of perfused capillaries available for exchange. This vascular area is dynamically affected by different factors, such as composition of the peritoneal fluid, the fill volume, and the production of inflammatory agents. Peritoneal dialysis fluids that will be developed in the future for children should allow an optimization of the fill volume owing to a better tolerance in terms of lower achieved intraperitoneal pressure for a given fill volume. Moreover, future peritoneal dialysis fluids should protect the peritoneal membrane from hyperperfusion (lower glucose degradation products). PMID- 16048281 TI - Sodium sieving in children. AB - Sodium sieving is a consequence of dissociation between the amount of water and sodium transported over the peritoneal membrane. This dissociation occurs in the presence of aquaporin-mediated water transport. Sieving of sodium can be used as a rough measure for aquaporin-mediated water transport. Icodextrin contains glucose polymers, inducing ultrafiltration by colloid osmosis. Therefore, aquaporins play a minor role in ultrafiltration, which is confirmed by the absence of sodium sieving. Icodextrin is very suitable for the daytime dwell in children on a nightly intermittent peritoneal dialysis regimen. Ultrafiltration obtained with icodextrin is similar to ultrafiltration obtained with 3.86% glucose after a 12-hour dwell. When using icodextrin in children, it is also confirmed by the absence of sodium sieving that the aquaporins play a minor role in ultrafiltration. PMID- 16048282 TI - Nutrition in children with kidney disease: pitfalls of popular assessment methods. AB - Children with chronic kidney disease (CKD) are considered at high risk for protein-energy malnutrition. Clinical practice guidelines generally recommend an evaluation of numerous nutritional parameters to give a complete and accurate picture of nutritional status. This review summarizes the potential limitations of commonly used methods of nutritional assessmentin the setting of CKD. Unrecognized fluid overload and inappropriate normalization of body composition measures are the most important factors leading to misinterpretation of the nutritional assessment in CKD. The importance of expressing body composition measures relative to height or height-age in a population in whom short stature and pubertal delay are highly prevalent is emphasized. The limitations of growth as a marker for nutritional status are also addressed. In addition, the prevailing belief that children with CKD are at high risk for malnutrition is challenged. PMID- 16048283 TI - Peritoneal dialysis training: a multisensory approach. AB - Undertaking peritoneal dialysis (PD) therapy poses a challenge to all patients with renal failure. The potentially high risk of infection makes it essential that patients undertaking PD have adequate training and ongoing support. Over recent years, increasing numbers of elderly patients, patients with significant learning disabilities, and patients with marked comorbidities have been accepted onto renal replacement therapy programs. For those undertaking PD in particular, this has posed new educational challenges. The Community Dialysis Team recognized an area of weakness in their current training program for these patient groups. The degree of literacy skills as well as the volume of written material and the amount of medical terminology used did not result in a user-friendly training program. A collaborative approach involving various members of the multidisciplinary team designed an appropriate training program for patients with learning disabilities. The new program included (1) a photographic bag-exchange procedure; (2) the provision of simple, step-by-step instructions on audiotape; (3) a new assessment sheet where words were replaced with symbols; (4) a redesigned daily record sheet (used to monitor clinical parameters); and (5) a simple contact card. The quality of the new training program was assessed by a small pilot study evaluation. The reduction of training times and the satisfactory peritonitis rates suggest that the new multisensory training program could be successfully implemented. The use of pictorial aids and more symbols, with less focus on the written word, made PD training a viable option for many individuals, including elderly patients and those with learning disabilities. The increased use of pictorial aids and symbols may also be helpful in training patients where there is a language barrier as well as the pediatric population. PMID- 16048284 TI - Longitudinal evaluation of a weight reduction program for patients on peritoneal dialysis. AB - OBJECTIVES: Patients on peritoneal dialysis (PD) are exposed to glucose-based dialysate solutions with consequent risk of obesity and its attendant health problems. We wished to examine the role of changes in dialysis prescription, individualized dietaryinput, and exercise on body weight and composition. DESIGN: A 1 year, prospective interventional study integrating the care of the renal nurse, dietitian, and physiotherapist to support, educate, and encourage overweight patients on PD in a weight-reduction program. PATIENTS: Patients were considered for the study if they had been on PD for more than 3 months, had a body mass index (BMI) > 25, and were considered medically fit to undergo the planned exercise program. Recruitment was intentionally limited to a maximum of 12 patients to facilitate group interaction. Weight, BMI, and bioimpedance were measured every 3 months. RESULTS: 8 of 11 enrolled patients completed the study; 3 received transplants. There was a significant fall in median body weight at initiation, from 94.6 kg to 92.4 kg at 6 months and 89.5 kg at 12 months (p = 0.017). This equates to a reduction in BMI from 33.2 (range 26.6 - 38.4) kg/m2 at initiation to 32.1 (range 24.5 - 37.6) kg/m2 at 6 months and 32.1 (range 23.9 - 36.5) kg/m2 at 12 months. There were no significant changes in total body water, lean body mass, or percentage body fat during the study. CONCLUSION: 7 of 8 patients achieved significant weight loss during the study. The use of an informal group setting motivated patients to continue with exercise and sensible eating patterns. This study demonstrates that, with adequate support, PD patients can achieve and maintain weight loss. PMID- 16048285 TI - Endoscopic management of pediatric brain tumors. AB - OBJECT: Primary endoscopic procedures for children with intraventricular brain tumors include endoscopic tumor biopsy and endoscopic tumor removal. The simultaneous treatment of hydrocephalus with endoscopic third ventriculostomy (ETV) or endoscopic septostomy increases the appeal of a minimally invasive endoscopic approach. METHODS: Eighty-five patients who underwent endoscopic management of an intraventricular brain tumor were identified from a prospective database. Of these patients, 26 were younger than 21 years of age at the time of diagnosis. The surgical technique, its success rate, and patient outcome were assessed. Illustrative cases are used in this study to detail the procedure of endoscopic tumor biopsy and resection. Endoscopic tumor procedures were successful in 96% of cases (23 of 24 endoscopic tumor biopsy samples and both endoscopic tumor removals). Fourteen simultaneous procedures were performed to treat hydrocephalus successfully. There was no recognized morbidity from the surgical procedures. CONCLUSIONS: Endoscopic surgery in children with intraventricular brain tumors is an effective and safe method for sampling of the lesion and, in select cases, its resection. This minimally invasive technique should be considered in situations in which the patient might thereby avoid a more conventional procedure, given the high rate of success and low morbidity associated with endoscopic management. PMID- 16048286 TI - Subfrontal transbasal approach and technique for resection of craniopharyngioma. AB - A multimodality approach to craniopharyngioma, including minimally invasive stereotactic techniques, microsurgery, conventional radiotherapy, and radiosurgery has been recommended to tackle craniopharyngioma aggressively while minimizing harm to the patient. With all approaches, there are varying levels of risk for endocrinological morbidity, vascular complications, neuropsychological and behavioral disorders, neurocognitive disorders, and learning disabilities. Although many treatment options are available, total tumor resection remains the most commonly performed procedure for treatment of craniopharyngioma, and it is still believed to give the patient the greatest chance of having an independent and productive life with low risk of recurrences. The authors prefer the subfrontal transbasal approach for resection of these tumors, and they describe this approach and illustrate it with the accompanying figures. PMID- 16048287 TI - A novel brainstem tumor model: guide screw technology with functional, radiological, and histopathological characterization. AB - OBJECT: Survival rates for high-grade brainstem tumors are approximately 10% and optimal therapy has yet to be determined. Development of a satisfactory brainstem tumor model is necessary for testing new therapeutic paradigms that may prolong survival. The authors report the technique, functional progression, radiological appearance, and histopathological features of a novel brainstem tumor model in rats. METHODS: Thirty female Fischer 344 rats were randomized (10 animals/group) to receive an injection of either 3 ml of 9L gliosarcoma cells (100,000 cells), 3 ml of F98 glioma cells (100,000 cells), or 3 ml of medium (Dulbecco modified Eagle medium) into the pontine tegmentum of the brainstem. Using a cannulated guide screw system implanted in the skull of the animal, rats in each group were injected at coordinates 1.4 mm to the right of the sagittal and 1 mm anterior to the lambdoid sutures, at a depth of 7 mm from the dura mater. The angle of the syringe during injection was anteflexed 5 degrees from the vertical. Postoperatively, the rats were evaluated for neurological deficits by using an automated rotarod test. High-resolution [18F]fluorodeoxyglucose-positron emission tomography (FDG-PET) fused with computerized tomography (CT) scans were acquired pre- and postoperatively through the onset of hemiparesis and correlated accordingly. Kaplan-Meier curves were generated for survival and disease progression, and brains were processed postmortem for histopathological investigation. The 9L and F98 tumor cells grew in 95% of the animals in which they were injected and resulted in a statistically significant mean onset of hemiparesis of 16.5 6 0.56 days (p = 0.001, log-rank test), compared with animals in the control group, which had no neurological deficits by Day 45. The FDG-PET studies coregistered with CT scans demonstrated space-occupying brainstem lesions, and this finding was confirmed by histological studies. Animals in the control group showed no functional, radiological, or pathological signs of tumor. CONCLUSIONS: Progression to hemiparesis was consistent in all tumor-injected animals, with predictable onset of symptoms occurring approximately 17 days postsurgery. The histopathological and radiological characteristics of the 9L and F98 brainstem tumors were comparable to those of aggressive primary human brainstem tumors. Establishment of this animal tumor model will facilitate the testing of new therapeutic paradigms for the treatment of these lesions. PMID- 16048288 TI - Endoscopic third ventriculostomy for hydrocephalus associated with tectal gliomas. AB - OBJECT: Tectal gliomas are a distinct form of pediatric brainstem tumor that present in patients with symptoms related to increased intracranial pressure due to obstructive hydrocephalus. The natural history of these lesions is often uniquely indolent. Thus, initial surgical therapies are directed at treatment of hydrocephalus, usually with ventricular shunt placement. Recently, third ventriculostomy has been used in patients with tectal gliomas, both as an initial procedure and after shunt failures. In this report the authors review their experience with the treatment of hydrocephalus in patients with tectal gliomas. METHODS: The authors reviewed 31 consecutive cases of tectal gliomas and compared the success rates of ventricular shunt placement with the success rates of endoscopic third ventriculostomy (ETV). Shunt placement procedures were associated with a significant number of malfunctions, and most patients required shunt revisions. The ETV procedure was attempted both as an initial treatment and after shunt malfunction. Overall, ETV was attempted in 18 patients and was performed successfully in all cases. At the time of follow-up evaluation, 16 patients (89%) were shunt free. CONCLUSIONS: The authors found that ETV could be performed with good long-term success both as an initial treatment and after shunt failure. Overall, ETV was found to be superior to ventricular shunt placement in the management of hydrocephalus associated with tectal gliomas. PMID- 16048289 TI - Neuroendoscopic resection of posterior third ventricular ependymoma. Case report. AB - The practice of neuroendoscopy in the definitive management of cystic tumors and hydrocephalus has been well established. Resection of solid intraventricular tumors by a primary endoscopic technique, however, has rarely been demonstrated. The authors present the case of a 31-year-old woman in whom endoscopic resection of a posterior third ventricular ependymoma was successfully accomplished. Metastatic workup yielded no sites of dissemination, adjuvant radiation therapy was deferred, and the patient has been without radiographic evidence of disease after 6 months of follow up. Endoscopic resection of solid tumors appears feasible in select patients and warrants further evaluation. PMID- 16048290 TI - Differentiation of choroid plexus tumors by advanced magnetic resonance spectroscopy. AB - OBJECT: The management of pediatric intraventricular tumors is highly dependent on identification of the tumor type. Choroid plexus papillomas, a common intraventricular tumor in children, can be difficult to distinguish radiographically from choroid plexus carcinomas and other common pediatric central nervous system (CNS) tumors. In this study to overcome the limitations of current noninvasive imaging modalities, the authors use novel magnetic resonance (MR) spectroscopy techniques in vivo to elucidate the identifying biochemical features of choroid plexus tumors that may facilitate diagnosis and treatment. METHODS: Based on an Internal Review Board-approved protocol, six children with newly diagnosed, untreated intraventricular brain tumors were identified. On retrospective review, this series included three choroid plexus papillomas and three choroid plexus carcinomas. Single-voxel proton MR spectroscopy with a short echo time was performed, and absolute metabolite concentrations (in mmol/kg) were determined using fully automated quantitation. These results were compared with MR spectroscopy profiles obtained in 54 other untreated CNS neoplasms in children. The myo-inositol (mI) level was significantly higher in choroid plexus papillomas (> 10 mmol/kg), uniquely distinguishing these tumors from choroid plexus carcinomas and all other tumors. Choroid plexus carcinomas, on the other hand, had significantly elevated levels of choline when compared with choroid plexus papillomas. CONCLUSIONS: In this study the authors find that mI is a biochemical constituent that uniquely identifies choroid plexus papillomas and can be used as a noninvasive means of diagnosis and for follow-up evaluations in patients with this disease. PMID- 16048291 TI - Surgical outcomes and seizure control rates after resection of dysembryoplastic neuroepithelial tumors. AB - OBJECT: In this study the authors review the outcomes in pediatric patients who presented with seizures and underwent resection of dysembryoplastic neuroepithelial tumors (DNETs). The authors focus on the diagnostic evaluation and surgical techniques that facilitate gross-total tumor resection and subsequent freedom from seizures. METHODS: Eighteen patients between the ages of 1 month and 13 years who presented with seizures underwent resection of DNETs between January 1992 and December 2004. Preoperative evaluation included magnetic resonance (MR) imaging and interictal scalp electroencephalography (EEG) in all patients, functional MR imaging in eight patients, video monitoring with ictal scalp EEG in 12 patients, interictal single-photon emission computerized tomography (SPECT) scanning in one patient, and ictal SPECT scanning in two patients. Thirteen patients underwent one-stage procedures, whereas five underwent two-stage procedures (implantation of monitoring electrodes followed by tumor resection), either for functional language mapping (three patients) or due to inconclusive preoperative data (two patients). Intraoperative electrocorticography (ECoG) was performed in 17 patients and led to resection of the cerebral cortex beyond the tumor margins in 10 of them. According to operative reports, gross-total tumor resections were achieved in all patients, but one child had minimal residual tumor on postoperative MR images that has remained stable. The only surgical complication was a transient third cranial nerve palsy. Over a median follow-up duration of 1.6 years, all patients are seizure free and without radiographically detected tumor recurrence. CONCLUSIONS: Dysembryoplastic neuroepithelial tumors are a highly treatable cause of epilepsy in children. Excellent rates of complete tumor resection and seizure control with minimal morbidity can be attained using intraoperative ECoG and two-stage surgical procedures when appropriate. PMID- 16048292 TI - Pediatric sellar tumors: diagnostic procedures and management. AB - The diagnosis and management of pediatric sellar lesions is discussed in this paper. Craniopharyngiomas account for the majority of pediatric sellar masses, and pituitary adenomas are extremely uncommon during childhood. The diagnosis of sellar lesions involves a multidisciplinary effort, and detailed endocrinological, ophthalmological, and neurological testing is critical in the evaluation of a new sellar mass. The management of pituitary adenomas varies depending on the entity. For most tumors other than prolactinomas, transsphenoidal resection remains the mainstay of treatment. Less invasive methods, such as endoscopic transsphenoidal surgery and stereotactic radiosurgery, have shown promise as primary and adjuvant treatment modalities, respectively. PMID- 16048293 TI - Pilomyxoid astrocytoma: diagnosis, prognosis, and management. AB - Pilomyxoid astrocytoma (PMA) is a recently defined pediatric brain tumor; PMAs were previously classified within the pilocytic astrocytoma (PA) category. Nevertheless, PMA has different histological features and has been shown to behave more aggressively than PA. These findings indicate that PMA may be a unique entity that is distinct from PA, or it may be an unusual variant. To increase awareness of PMA within the neurosurgical community, the authors review the diagnostic criteria, prognostic implications, and current management of this recently described pediatric low-grade astrocytoma. PMID- 16048294 TI - Atypical teratoid/rhabdoid tumors of the central nervous system: management and outcomes. AB - OBJECT: Atypical teratoid/rhabdoid tumors (ATRTs) represent a relatively newly categorized neoplastic entity. They commonly present in childhood, and have a rapidly progressive clinical course with a survival time of less than 1 year. Treatment regimens have been nonuniform. In this retrospective review of patients with ATRTs who were treated at the authors' institution according to a uniform protocol, the goal was to assess the efficacy of the treatment and its outcome. METHODS: Over a 7-year period, ATRT was diagnosed in 11 patients (six boys and five girls). The median age of the patients was 61 months, and their ages ranged from 3 months to 17 years. Signs and symptoms began, on average, a little more than 1 month before diagnosis and included the following: headache (36%), nausea and vomiting (46%), lethargy (18%), seizures (27%), cranial nerve findings (46%), ataxia (18%), long tract findings (18%), and hydrocephalus (46%). Tumor location was cortical in four patients, in the pineal region in four, in the posterior fossa in two, and spinal in one. In one patient disseminated disease was revealed on the initial imaging study; seven patients had disseminated tumor subsequently. Treatment consisted of chemotherapy in 11 patients, chemotherapy and local radiation in five, and chemotherapy and craniospinal radiation in three. Six patients are alive, three have died, and two were lost to follow-up review. The mean time to death was 24 months, and ranged from 2 to 67 months. Among the surviving patients the mean duration of follow up is 18.5 months and ranges from 2 to 37 months. The median time to progression was 3.5 months. CONCLUSIONS: Atypical teratoid/rhabdoid tumors are malignant lesions with rapid progression. Further study is necessary to determine the efficacy of therapy. PMID- 16048295 TI - Cranial base strategies for resection of craniopharyngioma in children. AB - The optimal treatment of craniopharyngioma in children remains a challenge. The use of complete excision to minimize recurrence continues to be controversial because of the risk of postoperative morbidity and death. Advances in skull base approaches, modern microsurgical techniques, neuroimaging, and hormone replacement therapy, however, have allowed safe gross- or near-total resection in the majority of cases. Total removal of these tumors, if possible, offers the best chance of cure for the patient. Although craniopharyngiomas are not strictly tumors of skull base origin, their intimate relationship with the neurovascular structures of this region often requires a skull base approach to maximize the surgical corridor and facilitate adequate microsurgical resection. In this review, the authors focus on commonly used skull base approaches for the surgical management of craniopharyngioma. They discuss the relative indications, advantages, disadvantages, and complications associated with each approach. Illustrative cases and intraoperative videos are presented. PMID- 16048296 TI - Surgical anatomy of supratentorial midline lesions. AB - OBJECT: In this paper the authors correlate the surgical aspects of deep median and paramedian supratentorial lesions with the connective fiber systems of the white matter of the brain. METHODS: The cerebral hemispheres of 10 cadaveric brains were dissected in a mediolateral direction by using the fiber dissection technique, corresponding to the surgical approach. CONCLUSIONS: This study illuminates the delicacy of the intertwined and stratified fiber laminae of the white matter, and establishes that these structures can be preserved at surgical exploration in patients. PMID- 16048297 TI - The anterior sylvian point and the suprasylvian operculum. AB - OBJECT: The sylvian fissure or lateral sulcus is the most identifiable feature of the superolateral brain surface and constitutes the main microneurosurgical corridor, given the high frequency of approachable intracranial lesions through this route. The anterior sylvian point (ASyP) divides this fissure in its main anterior and posterior rami and was evaluated in this study for its morphology, exact location, and sulcal and neural relationships to assess its suitability as an initial, visually identifiable landmark for further neuroimaging and intraoperative estimation of its adjoining suprasylvian structures. METHODS: This study is based on 32 formalin-fixed cerebral hemispheres. The brains were removed from the skulls of 16 cadavers after the introduction of plastic catheters through properly positioned burr holes; the number of specimens for some of the analyzed data differed because of incorrect positioning of catheters or damage to the studied structures caused by the initial steps of the study. The ASyP had a cisternal aspect in 94% of the specimens and was always located inferior to the triangular part of the inferior frontal gyrus, 2.3 6 0.5 cm in front of the inferior rolandic point. The ASyP was located underneath the 1.5-cm-diameter cranial area of the anterior aspect of the squamous suture. Its adjoining structures that compose the suprasylvian operculum have constant basic morphological configurations. CONCLUSIONS: The ASyP underlies the anterior aspect of squamous suture just behind the pterion, can be easily recognized, and constitutes a reliable initial sulcal landmark for further estimation of the suprasylvian sulcal and gyral structures. The suprasylvian operculum can be understood as a series of convolutions roughly arranged as a V-shaped convolution, with its vertex constituted by the ASyP, followed by three U-shaped convolutions and one C-shaped convolution. PMID- 16048298 TI - An anteromedial approach to the temporal horn to avoid injury to the optic radiation fibers and uncinate fasciculus: anatomical and technical note. AB - OBJECT: The aim of this study was to define an anteromedial approach to the temporal horn via a transsylvian approach to avoid injury to the optic radiation fibers as well as the uncinate fasciculus. This route was compared with standard surgical approaches to the temporal horn, and their relationship to the optic radiation and uncinate fasciculus was reviewed. METHODS: Three cadaveric brain specimens were prepared with freezing and thawing cycles according to the Klingler technique. Dissection was performed in a lateral-to-medial fashion with the help of wooden spatulas. Photographs were taken through the operating microscope at every level of the dissection. The dissection was continued until the optic radiation was encountered. Particular attention was paid to the relationship of the uncinate fasciculus with the optic radiation. An anteromedial transsylvian approach was defined to enter the temporal horn without injuring the optic radiation or the uncinate fasciculus. CONCLUSIONS: A transsylvian anteromedial approach through the pyriform cortex at the level of the anterior and superior surface of the uncus enables a safe entry into the temporal horn without injury to the optic radiation fibers or the main part of the uncinate fasciculus. PMID- 16048299 TI - Instructional video. Dissection of the human brain. PMID- 16048300 TI - Synthesis of a new chiral N,N,N-tridentate pyridinebisimidazoline ligand library and its application in Ru-catalyzed asymmetric epoxidation. AB - A small ligand library of chiral tridentate N,N,N-pyridinebisimidazolines have been synthesized for the first time. This new class of ligands can be easily tuned and synthesized on multi g-scale. The usefulness of the ligands is shown in the ruthenium-catalyzed asymmetric epoxidation with hydrogen peroxide as oxidant. Excellent yields (>99%) and good enantioselectivities (up to 71% ee) have been obtained for the epoxidation of aromatic olefins. [reaction: see text] PMID- 16048301 TI - Characterization of porosity in organic and metal-organic macrocycles by hyperpolarized 129Xe NMR spectroscopy. AB - Hyperpolarized (129)Xe NMR spectroscopy is used to establish the solid-state porosity of shape-persistent macrocycles with either an organic or metal-organic framework. These studies show that even upon removal of cocrystallized solvent molecules, the macrocycles maintain a porous or channeled structure. The technique can provide valuable information about systems for which X-ray crystallographic analysis is not feasible. [structure: see text] PMID- 16048302 TI - Stilbene-bridged tert-butylcalix[4]arene as photoswitchable molecular receptors. AB - Using a designed control of conformations of tert-butylcalix[4]arene between cone and pinched cone, a highly selective receptor for small electron-deficient molecules with photoswitching binding ability was attained. [reaction: see text] PMID- 16048303 TI - Asymmetric hydrogenation of N-sulfonylated-alpha-dehydroamino acids: toward the synthesis of an anthrax lethal factor inhibitor. AB - A novel and highly enantioselective Ru-catalyzed hydrogenation of N-sulfonylated alpha-dehydroamino acids has been discovered and demonstrated in the synthesis of an anthrax lethal factor inhibitor (LFI). Herein, this methodology is used to prepare N-sulfonylated amino acids in up to 98% ee. This unprecedented hydrogenation uses a chiral Ru catalyst rather than Rh as typical for acylated dehydroamino acids and esters, and this work reports the first asymmetric hydrogenation of a tetrasubstituted dehydroamino acid derivative using a Ru catalyst. [reaction: see text] PMID- 16048304 TI - Enhanced photovoltaic response in hydrogen-bonded all-organic devices. AB - Straightforward synthetic methodologies are reported for the functionalization of oligothiophenes with hydrogen-bonding motifs. Codeposition from a solution of symmetric melamine-terminated electron-donor oligomers with a complementary barbiturate-labeled electron-acceptor fullerene resulted in homogeneous films. Incorporation into photovoltaic devices gave a 2.5-fold enhancement in light energy to electrical energy conversion when compared to analogous systems with the non-hydrogen-bonding parent C(60). [reaction: see text] PMID- 16048305 TI - Cofacial porphyrin-ferrocene dyads and a new class of conjugated porphyrin. AB - A porphyrin-ferrocene dyad has been synthesized in which there is close face-to face contact between the two aromatic systems, providing a model for heterobimetallic polymers based on the same repeating unit. Attempts to synthesize the 2:1 adduct instead led to a remarkable intramolecular Heck-type cyclization which planarizes the system and extends the conjugation. [structure: see text] PMID- 16048306 TI - Dramatically enhanced carbon acidity of the nitrobenzyl fragment in a nickel(II) scorpionate complex. AB - The -CH(2)- group of the 2-nitrobenzyl pendant arm of the scorpionate complex I deprotonates in basic aqueous solution (pK(a) = 10.6), due to the coordination of the nitronate group to the nickel(II) center. Metal coordination enhances 2 nitrobenzene acidity by 10 orders of magnitude. [reaction: see text] PMID- 16048307 TI - Rapid synthesis of the N-methylwelwitindolinone skeleton. AB - An efficient, convergent synthesis of the core bicyclo[4.3.1]decane ring system of welwitindolinones is described. Key steps in the synthesis include an intramolecular palladium-catalyzed enolate arylation reaction to create the desired bicyclic skeleton and a Curtius rearrangement to install the bridgehead isocyanate unit. [reaction: see text] PMID- 16048308 TI - An electrochemical approach to the guanacastepenes. AB - An asymmetric approach toward the [6-7-5] ring system of the guanacastepenes is described. [structure: see text] PMID- 16048309 TI - An efficient synthesis of gougerotin and related analogues using solid- and solution-phase methodology. AB - The first solid-phase synthesis of the natural product gougerotin has been accomplished. The synthetic route is versatile and allows for diversification at position C-4 of the heterocycle, C-6' of the sugar ring, and both residues of the peptidic moiety at N-4' in a parallel fashion. [structure: see text] PMID- 16048310 TI - Highly enantioselective synthesis of fluorinated gamma-amino alcohols through proline-catalyzed cross-Mannich reaction. AB - A new, simple route for the synthesis of fluorinated beta-alkyl gamma-amino alcohols in optically pure form in only two steps and featuring proline catalysis from inexpensive and readily available starting materials is described. The applied strategy allows for the introduction of diversity into both the beta fluoroalkyl and alpha-alkyl groups of these compounds. [reaction: see text] PMID- 16048311 TI - Enantioselective synthesis of a highly potent selective serotonin reuptake inhibitor. An application of imidazolidinone catalysis to the alkylation of indoles with an alpha,beta-disubstituted alpha,beta-unsaturated aldehyde. AB - The synthesis of the highly potent and selective serotonin reuptake inhibitor 1 (BMS-594726) is described. In the key construction step, an enantioselective alkylation of the indole nucleus with an alpha-branched alpha,beta-unsaturated aldehyde 7 was accomplished utilizing MacMillan's imidazolidinone catalyst 3b. A rationale is presented for the unexpected stereochemical result, as well as the novel reactivity of the alpha-branched substrate. [reaction: see text] PMID- 16048312 TI - Enantioselective fluorescent recognition of a soluble "supported" chiral acid: toward a new method for chiral catalyst screening. AB - The long-chain aliphatic-group-substituted mandelic acid 3c, which is soluble only in THF and insoluble in water and many polar/nonpolar organic solvents, has been synthesized. This unique solubility allows 3c to be easily isolated from reaction mixtures and makes it potentially useful for catalyst screening. The fluorescent sensors (R)- and (S)-1 can be used to determine the ees of various samples of 3c generated from a series of catalyst screening experiments. The fluorescence measurements correlate well with the conventional HPLC-chiral column analysis. This work demonstrates that the enantioselective fluorescent recognition of organic substrates can lead to a fundamentally new method for chiral catalyst screening. [reaction: see text] PMID- 16048313 TI - Kinetic resolution of alcohols using a 1,2-dihydroimidazo[1,2-a]quinoline enantioselective acylation catalyst. AB - A new enantioselective acylation catalyst (Cl-PIQ), designed to provide enhanced pi-stacking with benzylic and cinnamyl alcohol substrates, was found to give considerably increased reaction rates and selectivities compared with the first generation catalyst CF(3)-PIP. [reaction: see text] PMID- 16048314 TI - Merrifield resin-supported chain transfer agents, precursors for RAFT polymerization. AB - The modification of Merrifield resins to form chain transfer agent (CTA) precursors for reversible addition fragmentation chain transfer (RAFT) polymerization is investigated. A series of CTA precursor resins were prepared and characterized by FTIR and elemental analysis (EA). [reaction: see text] PMID- 16048315 TI - Synthesis of vinylogous carbamates by rhodium(II)-catalyzed olefination of tertiary formamides with a silylated diazoester. AB - Treatment of tertiary formamides with a silylated diazoester in the presence of a rhodium(II) catalyst leads to the formation of 3-amino-2-silyloxyacrylates in good yield. No olefination is observed if a nonsilylated diazo compound is employed. [reaction: see text] PMID- 16048316 TI - Why downfield proton chemical shifts are not reliable aromaticity indicators. AB - Traces of magnetizability, traces of magnetic shielding at the hydrogen nuclei, and nucleus-independent chemical shift are not reliable aromaticity quantifiers for planar conjugated hydrocarbons. A measure of aromaticity is provided by the out-of-plane tensor components, whose magnitude is influenced by the pi-ring currents. The failure of nucleus-independent chemical shift in this regard was proved for the molecule shown in the abstract graphic, sustaining a diatropic pi current. The validity of the ring-current model is reaffirmed. [structure: see text] PMID- 16048317 TI - An anionic nucleophilic catalyst system for the diastereoselective synthesis of trans-beta-lactams. AB - Trans-disubstituted beta-lactams show increasing utility and prominence in numerous pharmaceutical applications, making their asymmetric synthesis an attractive goal for chemists. We introduce an anionic, nucleophilic catalyst system that provides an efficient, diastereoselective route to trans disubstituted beta-lactams, a complement to our previously described catalytic methodology for generating the corresponding cis diastereomers. This catalytic, "switch mechanism" process allows for flexibility in the stereoselective synthesis of beta-lactams, producing either cis or trans products as desired from the same substrates. [reaction: see text] PMID- 16048318 TI - Synthesis of gamma-substituted peptide nucleic acids: a new place to attach fluorophores without affecting DNA binding. AB - Molecular beacon strategies using PNA are currently restricted to fluorophore attachment to the ends of the PNA. We report the synthesis of PNA oligomers wherein fluorophores can be attached to the PNA backbone from novel gamma-lysine PNA monomers. Oligomers incorporating the modified PNA showed comparable thermal stability to the corresponding aegPNA oligomer with DNA. When the modified PNA oligomer was annealed with complementary DNA, the fluorescence intensity increased 4-fold over the unbound PNA. [structure: see text] PMID- 16048319 TI - A rapid 1H NMR assay for enantiomeric excess of alpha-substituted aldehydes. AB - In situ derivatization of a variety of alpha-substituted aldehydes via reaction with chiral amines allows convenient and efficient determination of enantiomeric excess. (1)H NMR analysis of the imine diastereomer ratio can be conducted immediately after the aldehyde and amine have been mixed. The results correlate well with ee values determined by more traditional (and slower) methods. This approach may be broadly applicable to alpha-substituted aldehydes. [reaction: see text] PMID- 16048320 TI - Microwave-assisted intramolecular [2 + 2] Allenic cycloaddition reaction for the rapid assembly of bicyclo[4.2.0]octa-1,6-dienes and bicyclo[5.2.0]nona-1,7 dienes. AB - Microwave irradiation of alkynyl allenes affords an intramolecular [2 + 2] cycloaddition reaction. This cycloaddition provides an efficient route to bicyclomethylenecyclobutenes. The reaction occurs with complete regioselectivity for the distal double bond of the allene for the selective formation of a variety of hetero- and carbocyclic substrates. Bicyclo[4.2.0]octadienes and bicyclo[5.2.0]nonadienes have been prepared in high yield. [reaction: see text] PMID- 16048321 TI - A new RNA synthetic method with a 2'-O-(2-cyanoethoxymethyl) protecting group. AB - A novel method for the synthesis of RNA oligomers with 2-cyanoethoxymethyl (CEM) as the 2'-hydroxyl protecting group has been developed. The new method allows the synthesis of oligoribonucleotides with an efficiency and final purity comparable to that obtained in DNA synthesis. [structure: see text] PMID- 16048322 TI - Synthesis and cytotoxicity of novel sansalvamide A derivatives. AB - Described are the syntheses of 14 derivatives of the natural product Sansalvamide A, where two are more active against HCT 116 colon cancer cell lines than the natural product. These derivatives were synthesized using a combinatorial-type strategy that permits elucidation of the amino acid role in the cytotoxicity, and they lay the groundwork for development of new anticancer agents. [structure: see text] PMID- 16048323 TI - High-yield solution-phase synthesis of di- and trinucleotide blocks assisted by polymer-supported reagents. AB - A new solution-phase phosphoramidite approach is reported for oligonucleotide synthesis employing recyclable solid-supported reagents. It uses polyvinyl pyridinium tosylate as the activator of a nucleoside-3'-O-phosphoramidite in the coupling step with a 5'-OH nucleoside or dinucleotide. The resulting phosphite triester was either sulfurized or oxidized using polystyrene-bound trimethylammonium tetrathionate or periodiate. This method avoids complicated purification steps, as excess reagents are easily removed by filtration. [reaction: see text] PMID- 16048324 TI - On the [2,3] sigmatropic rearrangement of allylic nitro compounds. AB - Allylic nitro compounds undergo relatively clean [2,3] sigmatropic rearrangement upon heating in refluxing 1,2-dichlorobenzene in the presence of DABCO to give the corresponding allylic alcohols via the intermediate allylic nitrite. [reaction: see text] PMID- 16048325 TI - A total synthesis of (+/-)-trans-kumausyne. AB - A short total synthesis of (+/-)-trans-kumausyne is reported. Key steps include a tandem ring-opening-ring-closing metathesis and the effective introduction of the pentenyl side chain by allylation-cross metathesis. [reaction: see text] PMID- 16048326 TI - Synthesis of the bis-spiroacetal moiety of spirolides B and D. AB - An enantioselective synthesis of the bis-spiroacetal fragment of spirolides B and D is reported. The carbon framework was constructed via Barbier reaction of dihydropyran 3 with aldehyde 4, followed by a double oxidative radical cyclization to construct the bis-spiroacetal. A silyl-modified Prins cyclization and enantioselective crotylation successfully installed the stereocenters in the cyclization precursor. The initial unsaturated bis-spiroacetals 2a-d underwent equilibration during epoxidation to trans-epoxide 14 that was converted to a tertiary alcohol. [reaction: see text] PMID- 16048327 TI - The first regiospecific, enantiospecific total synthesis of 6-oxoalstophylline and an improved total synthesis of alstonerine and alstophylline as well as the bisindole alkaloid macralstonine. AB - A Wacker sequence has been modified to rapidly generate ring E of (-) alstonerine, (+)-6-oxoalstophylline, and (-)-alstophylline via a domino process. This one-pot sequence provides facile entry into the dihydropyranyl enone unit of many Alstonia alkaloids. [structure: see text] PMID- 16048328 TI - Single-step synthesis of D3h-symmetric bicyclooxacalixarenes. AB - Bicyclooxacalixarenes are obtained in up to 95% yield by condensation of phloroglucinol with electron-poor meta-dihalogenated benzenes or pyridines. Single-crystal X-ray analysis reveals that these compounds adopt highly symmetrical, all-1,3-alternate cage-like structures. [structure: see text] PMID- 16048329 TI - Lewis base catalyzed ring opening of aziridines with silylated nucleophiles. AB - The ring opening of N-tosylaziridines with trimethylsilylated nucleophiles, catalyzed by N,N,N',N'-tetramethylethylenediamine, led to the production of beta functionalized sulfonamides in good to excellent yields with high regioselectivity. [reaction: see text] PMID- 16048330 TI - Synthesis of a thermally stable hybrid acene-thiophene organic semiconductor via a soluble precursor. AB - An acene fused-thiophene hybrid p-semiconductor exhibiting high thermal stability has been synthesized via a soluble precursor bearing sterically interacting trimethylsilyl groups. [structure: see text] PMID- 16048331 TI - Recognition of anionic carbohydrates by an artificial receptor in water. AB - A tris-cationic artificial receptor 1 efficiently binds anionic carbohydrates even in aqueous solution as shown by NMR and UV titration experiments. Complex formation involves both ion pair formation and H-bonds to the sugar, explaining the preference for saccharides compared to simple anions and the observed selectivities among different sugars. [structure: see text] PMID- 16048332 TI - A rapid synthesis of hexofuranose-like iminosugars using ring-closing metathesis. AB - Two new 1-N-iminosugars have been prepared as hexofuranose analogues in an efficient manner by an RCM-based route. Both 3,4-disubstituted pyrrolidines display moderate inhibitory activity against Mycobacterium smegmatis galactan biosynthesis. [structure: see text] PMID- 16048333 TI - Enantioselective recognition of 1,2-amino alcohols by reversible formation of imines with resonance-assisted hydrogen bonds. AB - A chiral aldehyde with three H-bond donating groups (2) has been synthesized. This aldehyde binds a variety of chiral 1,2-amino alcohols in benzene with the same sense of stereoselectivity. Computational and experimental data indicate that one imine bond, one resonance-assisted H-bond to the imine nitrogen, and two H-bonds to the alcoholic oxygen all play an important role in the stereoselective recognition. [structure: see text] PMID- 16048334 TI - Using peptidic inhibitors to systematically probe the S1' site of caspase-3 and caspase-7. AB - Fifteen ketone-containing peptides were designed, synthesized, and used to probe the effect of substitution at the P1' position on caspase-3 and -7 inhibition. Even with the large bias of Ac-Asp-Glu-Val-Asp at the P4-P1 positions, certain peptides with cyclic functionality in the P1' position show a dramatically reduced ability to inhibit these caspases. Additionally, trends toward isozyme selectivity were also uncovered for particular P1' substituents. The data indicate that substitution in the P1' position can drastically affect both caspase inhibition and selectivity. [structure: see text] PMID- 16048335 TI - Catalytic enantioselective Diels-Alder reaction by self-assembly of the components on a Lewis acid template. AB - Thermal Diels-Alder reaction of 2,4-hexadienol with methyl acrylate is unselective. By simultaneous coordination of diene and dienophile to a chiral bimetallic Lewis acid catalyst, a LACASA-DA reaction occurs with complete control of regio-, diastereo-, and enantioselectivity to give a single adduct. [reaction: see text] PMID- 16048336 TI - Synthesis of homoallylic amines by hydrozirconation-imine addition of allenes. AB - Hydrozirconation of allenes followed by in situ transmetalation to dialkylzinc leads to the formation of an allylic zinc species that, upon addition of aldimines to the reaction mixture, provides homoallylic amines in 64-85% yield. [reaction: see text] PMID- 16048337 TI - ROBIA: a reaction prediction program. AB - A reaction prediction program, ROBIA, has been developed. This interactive computer program predicts the products of organic reactions from the starting materials and the reaction conditions, on the basis of the selected transformations within its database. This mechanistic approach generates a large number of products, from which the most important are selected using filters and molecular modeling calculations. The procedure has been applied to the possible biosynthesis pathway of dolabriferol. [reaction: see text] PMID- 16048338 TI - Photochemical release of aldehydes from alpha-acetoxy nitroveratryl ethers. AB - Photolabile aldehyde-releasing precursors (alpha-acetoxy ethers) were prepared by reduction of the corresponding esters with DIBAL and quenching the intermediate aluminum hemiacetal with acetic anhydride. These species smoothly released aldehydes upon irradiation with UV light at 350 nm. Using this method, not only simple model aliphatic aldehydes were liberated but also specimens relevant for the flavor and fragrance industry (methional, phenylacetaldehyde, and (R) citronellal). [reaction: see text] PMID- 16048339 TI - Pd-catalyzed tandem C-N/C-C coupling of gem-dihalovinyl systems: a modular synthesis of 2-substituted indoles. AB - 2-substituted indoles were synthesized via a Pd-catalyzed tandem C-N/Suzuki Miyaura coupling from readily prepared ortho-gem-dihalovinylanilines. Optimal conditions used a Pd(OAc)(2)/S-Phos catalyst in the presence of K(3)PO(4).H(2)O and an organoboron reagent, which included boronic acids, esters, alkyl 9-BBN derivatives, and trialkylboranes. Yields of the desired indoles were good to excellent using low catalyst loadings (typically 1 mol %). [reaction: see text] PMID- 16048340 TI - Oxidative cyclization reactions: amide trapping groups and the synthesis of furanones. AB - Intramolecular anodic coupling reactions involving ketene dithioacetal radical cations and amide trapping groups have been examined. The reactions generate furanone products and benefit greatly from the addition of water to the reaction medium. The cyclization reactions lead to products having stereochemistry that is directly analogous to oxidative cyclization reactions utilizing ketene dithioiacetal radical cations and alcohol trapping groups. [reaction: see text] PMID- 16048341 TI - Preparation of homoallylic homopropargylic alcohols from 2-vinyloxiranes. AB - Beta,gamma-unsaturated aldehydes generated in situ by treatment of 2 vinyloxiranes with a catalytic amount of Sc(OTf)(3) or BF.OEt(2) are effectively trapped by B-allenyl-9-BBN to afford homoallylic homopropargylic alcohols in high yield. An enantioselective version has been demonstrated, and a convenient synthesis of 9-allenyl-9-BBN is described. [reaction: see text] PMID- 16048342 TI - Cationic rhodium(I)/BINAP complex-catalyzed isomerization of secondary propargylic alcohols to alpha,beta-enones. AB - We have developed a cationic rhodium(I)/BINAP complex-catalyzed isomerization of secondary propargylic alcohols to alpha,beta-enones. The asymmetric variant of this reaction, a kinetic resolution of secondary propargylic alcohols, was also developed with good selectivity. The mechanistic study revealed that the isomerization proceeds through intramolecular 1,3- and 1,2-hydrogen migration pathways. [reaction: see text] PMID- 16048343 TI - Carbon-carbon and carbon-heteroatom bond formation on solid phase using cationic iron carbonyl complexes. AB - Iron-mediated methodology for the formation of carbon-carbon and carbon heteroatom sp(3) bonds on solid phase has been developed. Treatment of a polymer bound cationic iron cyclohexadienyl complex with carbon, oxygen, nitrogen, and phosphorus nucleophiles, followed by cleavage with amines and subsequent decomplexation, yielded 18 different cyclohexadienoic acid amides of high purity. [reaction: see text] PMID- 16048344 TI - A scalable route to trisubstituted (E)-vinyl bromides. AB - An effective, readily scalable two-step synthesis of trisubstituted (E)-vinyl bromides involving bromination of alpha,beta-unsaturated lactones followed by hydrolytic fragmentation has been developed. Several trisubstituted (E)-vinyl bromides, including multigram quantities of (+)-(E)-4-bromo-2-methyl-3-pentenol, a synthetic intermediate required for the C(8)-C(11) moieties of (+)-tedanolide (1) and (+)-13-deoxytedanolide (2), illustrate the utility of this protocol. [reaction: see text] PMID- 16048346 TI - Gallium-mediated allyl transfer from bulky homoallylic alcohol to aldehydes via retro-allylation: stereoselective synthesis of both erythro- and threo homoallylic alcohols. AB - Retro-allylation of bulky gallium homoallylic alkoxides occurs to generate (Z)- and (E)-crotylgallium reagents stereospecifically, starting from erythro- and threo-homoallylic alcohols, respectively. The (Z)- and (E)-crotylgallium reagents immediately reacted with aromatic aldehydes to afford the corresponding erythro- and threo-homoallylic alcohols, respectively. [reaction: see text] PMID- 16048345 TI - Conjugated polymer-based fluorescence turn-on sensor for nitric oxide. AB - A turn-on fluorescent sensor for NO (g) in solution was synthesized using a bipyridyl-substituted poly(p-phenylene vinylene) derivative (CP1) as the sensory scaffold. The action of NO (g) upon the CP1-Cu(II) complex reduces it to the CP1 Cu(I) complex with a concomitant 2.8-fold increase in emission intensity. The reagent is selective for NO (g) versus other biological reactive nitrogen species, except for nitroxyl, and has a detection sensitivity limit of 6.3 nM. [structure: see text] PMID- 16048347 TI - Benzothiazines in synthesis. A total synthesis of pseudopteroxazole. AB - An enantioselective total synthesis of the naturally occurring antitubercular agent pseudopteroxazole is described. The synthesis is organized around the use of a stereoselective, intramolecular addition of a sulfoximine carbanion to an alpha,beta-unsaturated ester to form an enantiomerically pure benzothiazine. Other important processes include a completely stereoselective intramolecular Friedel-Crafts alkylation and a stereoselective and regioselective hydrogenation. [structure: see text] PMID- 16048348 TI - Concise synthesis of all stereoisomers of beta-methoxytyrosine and determination of the absolute configuration of the residue in callipeltin A. AB - All stereoisomers of beta-methoxytyrosine (beta-OMeTyr), a stereo-undefined component of callipeltin A, were synthesized from L- and D-tyrosine. The stereochemistry of beta-OMeTyr in callipeltin A was determined to be 2R,3R by an oxidative procedure and Marfey's analysis. [structure: see text] PMID- 16048349 TI - Solution-phase synthesis with solid-state workup of an O-glycopeptide with a cluster of cancer-related T antigens. AB - An N-terminal glycopeptide of asialoglycophorin AM with three O-linked T antigens was prepared by "solution-phase synthesis with solid-state workup" using unprotected glycosyl amino acids as building blocks. For the glycopeptide assembly, all reactions were conducted in homogeneous NMP solutions, while the product of each reaction was readily isolated as solid precipitates upon addition of diethyl ether. In the preparation of building blocks, a robust approach was established to selectively alpha-glycosylate Ser and Thr derivatives. [reaction: see text] PMID- 16048350 TI - Diastereoselective intermolecular Pauson-Khand reactions of chiral cyclopropenes. AB - In this Letter, it is demonstrated that the unusual reactivity of cyclopropenes can increase the scope and utility of intermolecular Pauson-Khand reactions. The well-defined chiral environment of cyclopropenes has a powerful influence on the diastereoselectivity of the reactions and leads to the production of a single cyclopentenone in each of the described cases. The cyclopropane ring strongly influences the stereochemistry of reactions at the enone, and the three-membered ring can subsequently be cleaved under mild conditions. [reaction: see text] PMID- 16048351 TI - Selective Michael-aldol reaction by use of sterically hindered aluminum aryloxides as Lewis acids: an easy approach to cyclobutane amino acids. AB - A formal [2 + 2] cycloaddition of 2-amidoacrylates with monosubstituted donor olefins, including its asymmetric version, is described. The stereoselectivity of this reaction can be modulated by the use of sterically hindered aluminum aryloxides or methylaluminoxane as Lewis acids. The reaction was applied to the synthesis of both stereoisomers of 2-benzyloxycyclobutane-alpha-amino acid, which are protected serine analogues c(4)Ser(OBn). PMID- 16048352 TI - What are the real lessons from Vioxx? PMID- 16048353 TI - The effect of HMG-CoA reductase inhibitors on coenzyme Q10: possible biochemical/clinical implications. AB - The HMG-CoA reductase inhibitors, also known as statins, have an enviable safety profile; however, myotoxicity and to a lesser extent hepatotoxicity have been noted in some patients following treatment. Statins target several tissues, depending upon their lipophilicity, where they competitively inhibit HMG-CoA reductase, the rate-limiting enzyme for mevalonic acid synthesis and subsequently cholesterol biosynthesis. HMG-CoA reductase is also the first committed rate limiting step for the synthesis of a range of other compounds including steroid hormones and ubidecarenone (ubiquinone), otherwise known as coenzyme Q(10) (CoQ(10)). Recent interest has focused on the possible role CoQ(10) deficiency may have in the pathophysiology of the rare adverse effects of statin treatment. Currently, there is insufficient evidence from human studies to link statin therapy unequivocally to pathologically significantly decreased tissue CoQ(10) levels. Although statin treatment has been reported to lower plasma/serum CoQ(10) status, few human studies have assessed tissue CoQ(10) status. The plasma/serum CoQ(10) level is influenced by a number of physiological factors and, therefore, has limited value as a means of assessing intracellular CoQ(10) status. In those limited studies that have assessed the effect of statin treatment upon tissue CoQ(10) levels, none have shown evidence of a fall in CoQ(10) levels. This may reflect the doses of statins used, since many appear to have been used at doses below those recommended for their maximum therapeutic effects. Moreover, the poor bioavailability in those peripheral tissues tested may not reflect the effects the agents are having in liver and muscle, the tissues commonly affected in those patients who do not tolerate statins. This article reviews the biochemistry of CoQ(10), its role in cellular metabolism and the available evidence linking possible CoQ(10) deficiency to statin therapy. PMID- 16048354 TI - Undesirable effects of citrus juice on the pharmacokinetics of drugs: focus on recent studies. AB - It is well known that intake of grapefruit juice affects the pharmacokinetics of various kinds of drugs. It has been reported that other citrus juices also interact with certain drugs. To re-evaluate citrus juice-drug interactions based on currently available evidence, a literature search was conducted for new and updated information since the grapefruit juice-drug interaction was last reviewed in 1998. MEDLINE (1998-October 2004) was accessed and more than 200 reports were found. The effects of grapefruit juice ingestion on the pharmacokinetics of orally administered drugs have been reported for 40 drugs since the reviews published in 1998. Increases in either area under the concentration-time curve (AUC) or maximum plasma concentration (C(max)) were found with 34 of these, the major mechanism being considered to be inactivation of intestinal cytochrome P450 3A4, a so-called mechanism-based inhibition. Although recent reports point to the inhibitory effects of grapefruit juice on the function of P-glycoprotein, which transports substrates from enterocytes back into the lumen, the contribution to the bioavailability of drugs that are substrates of P-glycoprotein has not been established yet. Dramatic decreases in AUC and C(max) for two drugs in association with grapefruit juice ingestion has been reported and, in these cases, inhibitory effects on organic anion transporting polypeptide, which mediates absorption from the intestinal lumen to enterocytes, might be involved. Other citrus juices such as Seville (sour) orange juice and commonly ingested varieties of orange juice also showed significant effects on the AUC and C(max) of some drugs. Although the situation is complex and uncertainties remain, we recommend that patients avoid citrus juice intake while taking medications and that healthcare providers advise against citrus juice intake in this setting until any interactions with subject drugs can be clarified in clinical studies. PMID- 16048355 TI - Treating mood disorders during pregnancy: safety considerations. AB - Mood disorders in pregnancy may have a negative effect on self care and pregnancy outcome that affects the mother directly and the child indirectly. Thus, some women may require pharmacological treatment. Pharmacotherapy of mood disorders during pregnancy implies specific considerations. This paper presents an updated review of available studies on the treatment of mood disorders and present knowledge on teratogenicity, neonatal effects and long-term neurobehavioural effects for the different psychotropic drugs, including treatment with selective serotonin reuptake inhibitors (SSRIs), tricyclic antidepressants (TCAs), other antidepressants, benzodiazepines, lithium, carbamazepine/valproic acid, lamotrigine and novel antipsychotics. However, the existing knowledge on the use of antidepressants and mood stabilising agents during pregnancy is hampered by a lack of results from randomised controlled trials.SSRIs and TCAs have not been associated with an increased risk of major malformations, but poor neonatal adaptation has been described. Benzodiazepines used in the first trimester have been associated with orofacial clefts. Mood stabilisers such as lithium, carbamazepine and valproic acid (sodium valproate) are associated with an increased risk of fetal malformations. Both benzodiazepines and lithium may cause adaptation problems in the newborn. In utero exposure to novel antipsychotics has not been associated with congenital malformations; however, the data are still limited. The knowledge about long-term neurobehavioural effects in the offspring is still limited for all agents and requires further investigation. Possible adverse effects of fetal exposure must be balanced against the adverse effects of an untreated maternal mood disorder. PMID- 16048356 TI - Fetal safety of drugs used in the treatment of allergic rhinitis: a critical review. AB - Allergic rhinitis is the most common allergic disease. Pharmacological interventions are often not used in pregnancy because of alarming information in drug labels and patient information, even when evidence for safety exists.Low risk therapies could include immunotherapy, intranasal sodium cromoglycate (cromolyn sodium), beclometasone, budesonide and first-generation antihistamines. In a meta-analysis examining the safety of first-generation antihistamines in pregnancy, 200 000 first trimester exposures failed to show increased teratogenic risk. Loratadine is the most studied second-generation antihistamine (with a total patient cohort of 2147 women who were exposed) and does not appear to increase the risk of major congenital malformations; however, it has not been as well studied as the earlier antihistamines. Since desloratadine is the principal metabolite of loratadine, it can be assumed that a similar safety profile would fit for desloratadine as was described for loratadine although no direct human studies have been done. Decongestants have not been conclusively proven to affect the fetal outcome and may be used for short-term relief when no other safer alternatives are available. Intranasal corticosteroids have not been associated with an increase in congenital malformations in humans. Based on efficacy and the fact that there would be little systemic absorption, they can be considered a first-line treatment over oral antihistamines, decongestants and mast cell stabilisers; however, the number of controlled trials in pregnancy is limited. Intranasal corticosteroids are associated with minimal systemic effects in adults and are the most effective therapy for allergic rhinitis. Benefit-risk considerations must, therefore, be done but favour their first-line use during pregnancy. Because fetal safety is paramount, recommendations should be based both on the safety of the drugs during pregnancy and the comparative efficacy of the agent in the treatment of the underlying condition. This review exemplifies the fact that there are many safe treatment options for the clinician when dealing with allergic rhinitis during pregnancy. PMID- 16048357 TI - Benefit-risk assessment of raloxifene in postmenopausal osteoporosis. AB - Raloxifene, a nonsteroidal benzothiophene, is a second-generation selective estrogen receptor modulator (SERM) that is an antiresorptive agent. Raloxifene is a non-hormonal agent that binds to the estrogen receptor and results in estrogen agonist effects on bone and the cardiovascular system and estrogen antagonist effects on endometrial and breast tissue. Raloxifene has diverse pharmacodynamic properties due to its differential interactions with the estrogen receptor and tissue selectivity. Raloxifene was the first SERM to be approved for the prevention and treatment of postmenopausal osteoporosis. In this review, we conducted a systematic search of the literature for trials that evaluated the following outcomes: bone density, fractures, quality of life, cardiovascular outcomes, safety and adverse events. Raloxifene at the approved dosage of 60 mg/day increased lumbar spine bone density by 2.5% relative to control after 2 years of therapy. A large fracture prevention trial confirmed that treatment with raloxifene 60 mg/day for 3 years decreased the relative risk of incident vertebral fractures by 30-50% in women with prevalent fractures or osteoporosis. Extraskeletal effects of raloxifene include a reduction in total cholesterol and low density lipoprotein cholesterol levels. Assessment of the safety profile revealed that raloxifene was not associated with endometrial hyperplasia and that there was a 72% reduction in the incidence of invasive breast cancer in raloxifene-treated postmenopausal women with osteoporosis. Adverse events associated with raloxifene included an increase in the absolute risk of venous thromboembolism and an increase in the risk of hot flashes and leg cramps. In comparison to other osteoporosis therapies, raloxifene has a lesser impact on bone mineral density, a similar effect on the occurrence of vertebral fractures, but no effect on the frequency of non-vertebral fractures. Raloxifene can be recommended for the prevention of vertebral fractures in women with osteopenia/osteoporosis who are not at high risk of non-vertebral fractures and who do not have a past history of venous thromboembolism. PMID- 16048358 TI - Trends in spontaneous adverse drug reaction reports to the French pharmacovigilance system (1986-2001). AB - BACKGROUND: The French pharmacovigilance system is based on a network of 31 regional centres located in teaching hospitals and coordinated by the French Medicines Agency ("Agence Francaise de Securite Sanitaire des Produits de Sante" [Afssaps]). Since 1984, they have shared a common database of adverse drug reactions (ADRs) that are spontaneously reported by healthcare professionals. The objective of this study is to describe the characteristics of the reports and the reporting trends in the French pharmacovigilance spontaneous reporting database from 1986 to 2001. METHODS: All the reports from January 1986 to December 2001 were included. Drugs and ADRs were translated to anatomical therapeutic chemical (ATC) codes and MedDRA classifications, respectively. RESULTS: The total number of reports was 197 580 over the 16-year period, with linearly increase over time. The median (interquartile range [IQR]) age of the patients was 53 (34-70) and the male/female ratio was 0.82. The median (IQR) time between the date of occurrence of the ADR and the date of report was 73 days (34-166). The reporter was a specialist in 74% of the reports and a general practitioner in 17%. The annual rate of reporting according to medical demography strongly increased for the specialists, especially since 1994. At least one ADR was considered as serious in 44.8% of the reports. The ADRs were most frequently related to nervous system drugs (23%), followed by cardiovascular drugs (19%) and systemic anti-infectives (17%). The latter class had the fastest progression mostly due to antiretroviral therapy since 1996. According to the Medical Dictionary for Regulatory Activities (MedDRA) coding, the system organ most often reported was skin and subcutaneous tissue disorders (29%), followed by nervous system disorders (19%), gastrointestinal disorders (12%), blood and lymphatic system disorders (12%), vascular disorders (12%) and general disorders and administration site conditions (12%). DISCUSSION: All spontaneous reporting systems are affected by under reporting. One of their goals is to generate early signals, which might be more affected by reporting bias than by under-reporting. Some improvements should be made in the design of the French database, but data collected since 1986 constitute an essential tool for the routine work of the 31 pharmacovigilance centres. CONCLUSION: This first description of the data of the French pharmacovigilance database involving all drugs and ADRs shows an increasing tendency to reporting over time, especially in specialists and for systemic anti infective drugs. The database that uses hierarchical international classifications for drugs and adverse reactions may be used for further studies and could be the basis for an automatic signal generation system. PMID- 16048359 TI - Tolerability of treatments for postherpetic neuralgia. PMID- 16048361 TI - Older and wiser: fact or fiction? PMID- 16048362 TI - Caring through technological competency. PMID- 16048363 TI - Inaugural address: NASN's capitol investment for children. PMID- 16048364 TI - Understanding and managing neurally mediated syncope in the adolescent. AB - Syncope and near-syncopal symptoms are common events in the adolescent population. Syncope is defined as the transient loss of postural tone and consciousness with spontaneous recovery. Although most syncopal events are benign, they can generate extreme anxiety in the adolescent and his or her family. The re-occurrence of these events can have a negative impact on an adolescent's participation in daily activities. Understanding the process of assessment, diagnosis, and management of these teens can enable the school nurse to augment initial assessment and to support the prescribed treatment plan. This knowledge then can be used to help allay the adolescent's anxiety and to support efforts at self-management. This article explores current literature in regards to etiology, diagnosis, and medical treatment, and advocates a self-management protocol for neurally mediated syncope in the adolescent population with support of that plan by the school nurse. PMID- 16048365 TI - Caring for children with autism in the school setting. AB - Children with autism present unique challenges related to communication, behavior, and social skills. Each child with an autistic spectrum disorder (ASD) exhibits individual characteristics of the disorder. Early identification of autistic spectrum disorder has been shown to improve the child's benefit from educational interventions. There may be health issues related to autism including seizure disorders and dietary restrictions. Students with autism need special consideration when they come to the health office for screening, assessment of injuries, or administration of medication. Collaboration between educators and school nurses is needed to develop and implement Individual Education Plans and Individual Healthcare Plans. Community resources provide valuable assistance to families of children with this challenging disorder. PMID- 16048366 TI - The New Mexico school nurse and emergency medical services emergency preparedness course: program description and evaluation. AB - Illness and injuries are common among students and school staff. Therefore, school nurses must be prepared. In this study, a 16-hour scenario-based emergency preparedness course for school nurses was evaluated for its effectiveness. Effectiveness was measured by (a) traditional methods (written exams and confidence surveys) and (b) skills and performance evaluations in simulated emergencies called On-site Mock Emergency Scenarios. School nurses who completed the emergency preparedness course showed significant improvement in knowledge, confidence, and On-site Mock Emergency Scenarios scores that measured each nurse's ability to apply knowledge in simulated emergencies. PMID- 16048367 TI - The relationship between perceived and ideal body size and body mass index in 3rd grade low socioeconomic Hispanic children. AB - Very little is known about body satisfaction among minority children. This study examined the relationship between perceived and actual body size and Body Mass Index among 43 low-socioeconomic Hispanic 3rd-graders. Researchers measured participants' Body Mass Index; students self-reported Perceived Ideal Self Image and Perceived Actual Self Image using Collins' (1991) pictorial instrument scale of seven child body images that illustrate body weight from very thin to obese. The majority of students chose body images from the scale that depicted a healthy weight for both their Perceived Ideal Self Image and Perceived Actual Self Image. More boys than girls chose underweight as their Perceived Ideal Self Image. Thirty percent of the students were found to be overweight and 30 percent were at risk for being overweight. A small positive correlation between Perceived Actual Self Image and Body Mass Index was found. PMID- 16048368 TI - Smoking and depressive symptoms in a college population. AB - A number of studies have documented the relationship between smoking and depression in adolescent and adult populations. The purpose of this study was to examine the level of depressive symptoms among college-age smokers and to determine whether or not increases in cigarette use were associated with increases in the number of depressive symptoms. A non-experimental cross sectional design was used. Of the random sample (N = 895), 28% were current smokers, and more than 26% of the total sample reported high levels of depressive symptoms. Current smokers were more likely to report an increased level of depressive symptoms than nonsmokers were. Correlates of depressive symptoms included grade point average, marijuana use, and increased work hours. When controlling for these variables, the number of days smoked was predictive of depressive symptoms. School nurses have an important role in preventing smoking and treating smokers, as well as in preparing adolescents for the transition to college where many begin smoking. PMID- 16048369 TI - Schools' capacity to help low-income, minority children to manage asthma. AB - This article describes the challenges and strengths of asthma management in 14 low-income, predominantly African American urban elementary schools serving more than 5,000 students. Asthma prevalence was 24.5%. Teachers, school principals, parents, and children described how asthma was managed at school. Data from classmates of students with asthma showed that they had moderate to high levels of information about the disease. Data from teachers indicated the great need for practical instruction on how they might effectively support a child with asthma in the classroom and on the playground. Principals raised concerns about expectations for the functioning of school staff and implementation of school policies especially related to asthma emergencies. Parents reported a range of problems their children face at school. Data from children with asthma showed that 75% believed asthma affected their school work. Findings from this study should be useful to school personnel, health providers, and others who assist children and their families to manage asthma at school. Data suggest that making school nursing services available is warranted, given the impact of asthma on the school community. PMID- 16048370 TI - Data destruction. AB - School nurses are caretakers of a vast amount of sensitive student and family health information. In schools, older computer hardware that previously stored education records is recycled for less demanding student and employee functions. Sensitive data must be adequately erased before electronic storage devices are reassigned or are discarded. State and federal laws must be considered when permanently destroying personally identifiable student information. To fulfill their ethical and legal responsibilities, school nurses must be aware of record retention and data destruction policies and procedures for both paper and electronic records. PMID- 16048372 TI - Aging and gene expression in the primate brain. AB - It is well established that gene expression levels in many organisms change during the aging process, and the advent of DNA microarrays has allowed genome wide patterns of transcriptional changes associated with aging to be studied in both model organisms and various human tissues. Understanding the effects of aging on gene expression in the human brain is of particular interest, because of its relation to both normal and pathological neurodegeneration. Here we show that human cerebral cortex, human cerebellum, and chimpanzee cortex each undergo different patterns of age-related gene expression alterations. In humans, many more genes undergo consistent expression changes in the cortex than in the cerebellum; in chimpanzees, many genes change expression with age in cortex, but the pattern of changes in expression bears almost no resemblance to that of human cortex. These results demonstrate the diversity of aging patterns present within the human brain, as well as how rapidly genome-wide patterns of aging can evolve between species; they may also have implications for the oxidative free radical theory of aging, and help to improve our understanding of human neurodegenerative diseases. PMID- 16048373 TI - Risk-adjusted outcome analysis of endovascular abdominal aortic aneurysm repair in a large population: how do stent-grafts compare? AB - PURPOSE: To compare differences in the applicability and incidence of postoperative adverse events among stent-grafts used for repair of infrarenal aortic aneurysms. METHODS: An analysis of 6787 patients from the EUROSTAR Registry database was conducted to compare aneurysm morphological features, patient characteristics, and postoperative events for the AneuRx, EVT/Ancure, Excluder, Stentor, Talent, and Zenith devices versus the Vanguard device (control) and each other. Annual incidence rates of complications were determined, and risks were compared using the Cox proportional hazards analysis. RESULTS: The annual incidence rates were: device-related endoleak (types I and III) 6% (range 4%-10%), type II endoleak 5% (range 0.3%-11%), migration 3% (range 0.5%-5%), kinking 2% (range 1%-5%), occlusion 3% (range 1%-5%), rupture 0.5% (range 0%-1%), and all-cause mortality 7% (range 5%-8%). After adjustment for factors influencing outcome, AneuRx, Excluder, Talent, and Zenith devices were associated with a lower risk of migration, kinking, occlusion, and secondary intervention compared to the Vanguard device. Significant increased risk for conversion (EVT/Ancure) and reduced risk of aneurysm rupture (AneuRx and Zenith) and all-cause mortality (Excluder) were found compared to the Vanguard device. CONCLUSIONS: Significant differences exist between stent-grafts of different labels in terms of applicability and complications during intermediate to long term follow-up. Since each stent-graft has its drawbacks, no single label can be identified as the best. It is reassuring that developments in stent-grafts indeed result in better performance than the early stent-grafts. However, a single device incorporating all the perceived improvements should still be pursued. PMID- 16048374 TI - Endoscopic aneurysm sac fenestration as a treatment option for growing aneurysms due to type II endoleak or endotension. AB - PURPOSE: To evaluate endoscopic fenestration as a treatment option for growing aneurysm due to a type II endoleak or endotension after endovascular aneurysm repair (EVAR). METHODS: Eight patients (7 men; median age 69 years, range 55-79) who underwent "successful" EVAR were diagnosed with a growing aneurysm due to a type II endoleak (n=4) or endotension (n=4). Surgical intervention consisted of endoscopic fenestration of the sac and removal of all the thrombus material, preceded by clipping of the inferior mesenteric and all lumbar arteries in cases of endoleak. Fluid samples from the fenestrated aneurysm sac were analyzed for the presence of microorganisms and fibrin degradation products (FDP) and/or D dimers. RESULTS: The median duration of operation was 220 minutes (range 111 333). There was no perioperative mortality. In one patient, the endoscopic procedure was converted to an open fenestration procedure. Seven patients had uncomplicated follow-up and a clear decrease in the diameter of the sac; one patient was converted to open repair owing to continued sac growth despite fenestration. Bacterial cultures were negative in all patients, but high levels of FDP and/or D-dimers were found in all available samples, indicating continued fibrinolysis. CONCLUSION: Endoscopic fenestration, with or without endoscopic clipping of all side branches, seems to be an effective, reliable and minimally invasive treatment option for patients with a growing aneurysm due to type II endoleak or endotension. The high levels of FDP and/or D-dimers in the aneurysm sac are suggestive of hyperfibrinolysis, which may play an important role in aneurysm growth after EVAR. PMID- 16048375 TI - Flow visualization in a model of a bifurcated stent-graft. AB - PURPOSE: To use an in vitro flow model to investigate the flow patterns in a bifurcated stent-graft for abdominal aortic aneurysm (AAA) repair. METHODS: Experiments were performed in an in vitro test rig incorporating a simplified non planar model of an AAA. A two-component bifurcated device consisting of a stent structure and transparent polyurethane "graft" was deployed in the AAA model. Using a blood analogue fluid, a pulsatile blood flow waveform simulating resting flow condition was produced by means of a piston pump system. Flow patterns in the lateral and anteroposterior planes of the stent-graft were recorded and analyzed using flow visualization techniques. RESULTS: The flow patterns within the stent-graft were complex and influenced by the geometry of the stent-graft itself, as well as that of the aortic neck and iliac vessels. Regions of flow separation, low velocity and stagnation, and slow oscillatory flow near the walls were seen in the main body of the stent-graft. Constriction at the stump in the contralateral limb resulted in flow disturbances and flow separation. Kinking at the junctions of stent segments and folding of the graft compounded these complex flow structures. CONCLUSIONS: The flow structures within stent-grafts are complex, with features that may predispose to thrombus formation. Arterial geometry, including aortic neck angulation and iliac vessel tortuosity, and the design of the stent-graft are factors that influence hemodynamics and may impact the performance of aortic stent-grafts. PMID- 16048376 TI - Transcervical approach with protective flow reversal for carotid angioplasty and stenting. AB - PURPOSE: To report our initial experience using a transcervical approach for carotid angioplasty/stenting (CAS) that employs internal carotid artery (ICA) flow reversal for neuroprotection. METHODS: Seventeen patients (15 men; mean age 65 years, range 49-77) with significant carotid stenosis (mean 88%, 8 symptomatic) were treated with protected transcervical CAS. Eleven patients were considered at high risk for carotid endarterectomy; 8 were also considered high risk for transfemoral access (unfavorable aortic arch anatomy or advanced aortoiliac occlusive disease). Anesthesia was based on patient and anesthesiologist preferences. The approach consisted of a 2-cm cutdown over the common carotid artery and placement of a 9-F sheath. ICA flow was reversed and shunted into the jugular vein during the carotid intervention. RESULTS: Access and carotid stenting were successful in all cases. Thirteen procedures were performed under general and 4 under local anesthesia. Mean flow reversal time was 34+/-4 minutes (25 minutes in the last 7 cases). The patients tolerated the procedure well and had no neurological events. Four (23%) patients had significant oozing from the operative site; 2 developed small neck hematomas that were treated conservatively. All patients were discharged on the first postoperative day. There were no deaths, changes in neurological status, or restenosis over a mean follow-up of 12 months (range 1-24). CONCLUSIONS: Our initial experience demonstrates that a transcervical approach is a viable alternative for CAS. The procedure can be performed safely, with good initial clinical outcomes. The approach allows carotid flow reversal and emboli protection without introducing neuroprotection devices. The method appears best suited for patients at high risk for endarterectomy and transfemoral access. PMID- 16048377 TI - Bilateral brachial artery occlusion decreases internal carotid artery volume flow: a simple adjunct for cerebral protection? AB - PURPOSE: To investigate if a decrease in internal carotid artery (ICA) blood flow occurs with bilateral brachial artery occlusion (BBO), which may improve the effectiveness of cerebral protection devices during carotid interventions. METHODS: Thirty-two asymptomatic patients (21 men; mean age 67 years) with carotid atherosclerosis between 15% and 79% were enrolled in the study. Carotid duplex ultrasound was followed by volume flow rate (VF) determination in the right ICA, external carotid (ECA), and vertebral arteries. After baseline values were obtained, BBO was induced by bilateral arm pressure cuff inflation to 30 mmHg over the systolic pressure for no more than 3 minutes. VF measurements were repeated. RESULTS: Seventeen patients (responders) had an ICA VF decrease from 406+/-109 mL/min (+/-SD) to 303+/-90 mL/min (p=0.005), while 15 patients (nonresponders) had no significant change in their ICA VF (340+/-192 versus 447+/ 267 mL/min, p=0.22). In responders, ECA VF increased (190+/-65 to 232+/-125 mL/min), as did vertebral VF (77+/-53 to 95+/-60 mL/min; p>0.05). The ratio of ICA/ECA VF dropped from 2.13 to 1.31 in responders, but did not change in nonresponders. No patient exhibited any neurological symptoms during the study. Post cuff volume flows approximated baseline values. Cerebral magnetic resonance angiograms obtained in 10 responders revealed a complete circle of Willis in 8 (80%), while only 1 (16%) of 6 nonresponders had a complete pathway. CONCLUSIONS: A transient decrease in ICA VF, with concomitant elevations of the ECA and vertebral VFs, occurs with occlusion of the brachial arteries in the setting of a complete circle of Willis. Since no flow reversal occurs, this maneuver is insufficient to provide complete cerebral protection, but it may improve the effectiveness of cerebral protection devices and serve as an adjunctive maneuver in selected cases. Furthermore, changes in ICA VF may prove to be a noninvasive test for evaluating the integrity of the circle of Willis. PMID- 16048378 TI - Value of dual-phase multislice CT prior to minimally invasive therapy of iatrogenic renal injuries. AB - PURPOSE: To evaluate the diagnostic impact of multislice computed tomography (MSCT) in treatment planning prior to transarterial coil embolization of iatrogenic renal injuries. METHODS: Nine patients (median age 54 years, range 36 66) with iatrogenic renal vascular injury were treated with superselective coil embolization. Prior to therapy, a dual-phase (40 and 120 seconds) contrast enhanced MSCT was applied in 6 patients. Seven patients underwent renal ultrasonography. Multiplanar reconstructions of the MSCT scans were used to affirm ongoing bleeding and to localize the bleeding site at the level of the segmental or interlobar artery. MSCT and angiographic findings were compared to evaluate the accuracy of MSCT in the detection of the bleeding source prior to therapy. RESULTS: Multiplanar reconstructions of early-phase MSCT scans allowed precise identification of the bleeding interlobar or segmental artery in all 6 cases prior to angiography. In one case, MSCT was even able to detect a source of bleeding that was not revealed by selective renal angiography. CONCLUSIONS: Multiplanar reconstructions of MSCT data demonstrate not only the presence of hematoma but also confirm ongoing bleeding and the location of the feeder artery prior to minimally invasive therapy. PMID- 16048379 TI - CD151 promotes neovascularization and improves blood perfusion in a rat hind-limb ischemia model. AB - PURPOSE: To evaluate the efficiency of recombinant adeno-associated virus (rAAV) mediated CD151 gene delivery in promoting neovascularization and improving blood perfusion in the skeletal muscle of the rat hind-limb ischemia model. METHODS: CD151 was constructed into the rAAV vector. Twelve Wistar rats were randomly divided into 2 groups of 6 rats each and then intramuscularly injected with rAAV CD151 or rAAV-GFP, respectively, in one hind limb. Two weeks after gene delivery, the femoral arteries in the treated limbs were excised to establish the model of hind-limb ischemia. Expression of the transgene product CD151 was confirmed by Western blot and the reverse transcription polymerase chain reaction. The skin temperature, angiographic score, and capillary density of the hind limb were measured to assess blood perfusion and neovascularization 6 weeks after transfection. RESULTS: Compared to the group transfected with GFP, the CD151 group showed a 63% higher angiographic score (p<0.05) and an 18% increase in capillary density (p<0.05). In addition, the mean skin temperature of the AAV CD151-transfected hind limbs was equivalent to the level of the contralateral nonischemic limb, whereas the limb temperature in the GFP-transfected rats was significantly lower than the nonischemic control. The expression of CD151 in the ischemic hind limb injected with rAAV-CD151 was significantly higher than limbs injected with rAAV-GFP. The CD151 mRNA and protein expression was persistently observed in the injected muscle for at least 6 weeks after injection, while no human CD151 mRNA could be detected in remote organs or rAAV-GFP-injected muscles. CONCLUSIONS: This study demonstrates that the rAAV-mediated CD151 gene transfer into rat skeletal muscles is efficient, stable, and has no ectopic expression. Moreover, rAAV-mediated CD151 gene transfer stimulates neovascularization, especially arteriogenesis, and thereby improves blood perfusion in a rat hind limb ischemia model. These findings suggest that CD151 could be a new target for neovascularization therapy in ischemic disease, and rAAV-mediated CD151 gene transfer may be useful for treatment of ischemic disease. PMID- 16048380 TI - Preliminary investigation of mitoxantrone coating on stent-grafts to inhibit neointimal proliferation. AB - PURPOSE: To investigate the inhibition of neointimal proliferation induced by a stent-graft loaded with mitoxantrone. METHODS: Stent-grafts with and without mitoxantrone loading (150 microg per device) were inserted into the carotid artery of 7 and 6 rabbits, respectively. After an observation period of 28 days, the animals were sacrificed, and a detailed morphological and morphometric workup of the stented vessels was performed. RESULTS: Uncoated stent-grafts induced a thick neointima (median diameter 97+/-99 microm), whereas no neointima formed in mitoxantrone-loaded stent-grafts. However, the loaded devices were not fully covered by an endothelial layer. The underlying media was significantly thinner (31.8+/-5.6 versus 48.6+/-3.3 microm, p=0.002) and showed a widespread loss of smooth muscle cells. CONCLUSIONS: Mitoxantrone loading of a stent-graft inhibited the formation of a neointima. However, important regenerative processes were prevented as well, indicating a local overdose. More experiments using lower doses are warranted. PMID- 16048381 TI - Evaluation of a novel endoluminal vascular occlusion device in a porcine model: early and late follow-up. AB - PURPOSE: To evaluate the efficacy of a new vascular occlusion device (VOD) in a preclinical controlled study versus embolization coils. METHODS: The Biomerix VOD was made from a biodurable porous polyurethane matrix in the shape of a cylinder measuring 1.5 cm long by 6.0 mm wide. Thirty-three swine were selected to undergo embolization of a 3 to 5-mm-diameter iliac artery using either a single VOD (27 animals) or sufficient Cook fibered stainless steel coils to achieve angiographic occlusion (6 controls). Test animals were assigned to undergo angiography at 1 week (n=11), 1 month (n=6), 3 months (n=6), or 6 months (n=4). Two control animals were assigned to angiographic follow-up at 1 week, 1 month, and 3 months. Test and control animals were euthanized at each time point to explant occluded vessels for histological analysis. Study endpoints were device utilization, time to occlusion, postdeployment migration, and persistent angiographic occlusion at 7, 30, 90, and 180 days. RESULTS: One VOD was deployed in each test animal, whereas a mean 3.3+/-0.8 coils were needed to achieve angiographic occlusion in the 6 controls (p<0.001). The time to occlusion was significantly shorter with the VOD (1.46+/-0.73 versus 5.83+/-1.60 minutes for the coils; p<0.001). There was no evidence of recanalization or filling defects at the site of VOD deployment, while filling defects were seen in 3 of 6 coil-treated controls. The VOD arm showed superior angiographic occlusion versus coils at the 1-week, 1 month, and 3-month angiographic follow-up time points. Histological evaluation showed that the VOD was equivalent to the embolization coils at the 1-week (n=6) and 1-month (n=6) endpoint (100% luminal occlusion). In the 3-month group (n=6), the VOD showed 95% to 100% luminal occlusion versus 90% to 95% in the control animals. In the 6-month group, VOD showed 85% to 95% occlusion. CONCLUSIONS: The Biomerix VOD appears highly effective and reliable, resulting in significantly faster and longer lasting occlusion compared with fibered stainless steel embolization coils. PMID- 16048382 TI - Effect of stent oversizing on in-stent stenosis and lumen size in normal porcine veins. AB - PURPOSE: To present an in vivo evaluation of the effects of stent oversizing on in-stent stenosis and on actual lumen size in normal veins. METHODS: Twelve self expanding stents were implanted in the common iliac veins of 6 minipigs, which were divided in 2 groups for follow-up at 1 and 3 months, respectively. Oversizing ranged from 119.5% to 216% of the original vessel diameter. Intravascular ultrasound (IVUS) was used to measure vessel areas at the stent edges and midpoint. In-stent stenosis and decrease in actual lumen size were evaluated with IVUS immediately after implantation and during follow-up. RESULTS: There was a very strong positive linear regression between stent oversizing and in-stent stenosis in the 1-month model (r=0.947, p=0.004), in the 3-month model (r=0.931, p=0.007), and in the sum of these 2 groups (r=0.83, p<0.001). Regarding the change in actual lumen size, we found a strong negative linear regression with stent oversizing in the 1-month (r=-0.754, p=0.083) and the 3-month (r= 0.562, p=0.246) groups, but these did not attain statistical significance. However, a strong correlation between stent oversizing and a decrease in lumen size in all samples proved to be statistically significant (r=0.619, p=0.032). CONCLUSIONS: Stent oversizing was strongly correlated with increased in-stent stenosis and decreased actual lumen size at follow-up in normal non-stenotic porcine veins. PMID- 16048383 TI - Collapse of a stent-graft following treatment of a traumatic thoracic aortic rupture. AB - PURPOSE: To report a collapsed stent-graft used to treat a traumatic aortic rupture. CASE REPORT: A Gore TAG stent-graft was placed in a 20-year-old man with multiple injuries. Postimplantation computed tomographic angiography (CTA) demonstrated no contrast extravasation and total exclusion of the traumatic rupture. Routine CTA 3 months after implantation revealed a collapsed stent-graft located in the outer curve of the distal aortic arch. A Talent stent-graft was placed successfully within the collapsed prosthesis. Postimplantation CTA demonstrated no contrast extravasation and good apposition of the endograft to the aortic wall. At 6 months, the repair remains secure; there is no sign of graft collapse or endoleak. CONCLUSIONS: Collapse of stent-grafts can occur after treatment for traumatic aortic ruptures; endovascular methods can be used to restore a satisfactory luminal contour. PMID- 16048384 TI - Percutaneous transhepatic mechanical thrombectomy for acute mesenteric venous thrombosis. AB - PURPOSE: To present the management of symptomatic acute mesenteric venous thrombosis using a percutaneous thrombectomy device followed by resection of necrotic intestine. CASE REPORT: A 60-year-old woman developed acute abdomen and melena. Diffuse and extensive mesenteric and portal vein thromboses were diagnosed by computed tomography. Percutaneous transhepatic mechanical thrombectomy with an Oasis thrombectomy device removed approximately 80% of the thrombus in the portal and superior mesenteric veins. The patient underwent laparotomy immediately after thrombectomy, in which 100 cm of necrotic intestine was resected. Catheter-directed urokinase thrombolysis was performed for 3 days to address residual thrombi. The result was excellent, and the patient recovered without short bowel syndrome. CONCLUSIONS: The hydrodynamic thrombectomy system is a quick, reliable, efficient device that may offer an alternative to thrombolysis and surgical thrombectomy. Combining mechanical thrombectomy devices and surgery can be used to treat symptomatic acute mesenteric venous thrombosis. PMID- 16048385 TI - Endovascular repair of an aorto-left renal vein fistula due to a ruptured abdominal aortic aneurysm after EVAR. AB - PURPOSE: To report an unusual late complication of endovascular aneurysm repair: an arteriovenous fistula between the aneurysm sac and a retro-aortic left renal vein following sac expansion due to a type III endoleak. CASE REPORT: A 79-year old man developed an arteriovenous fistula between the aneurysm sac and a retro aortic left renal vein 67 months after endovascular aneurysm exclusion (EVAR). Aneurysm rupture was due to disconnection between the right iliac limb and an extender cuff. The problem was repaired percutaneously with another endograft bridging the two prostheses. At 16 months, the aneurysm sac diameter was decreased; there was no evidence of the AV fistula, and the patient was free from any complication related to the EVAR. CONCLUSIONS: This case emphasizes the need of close surveillance even in the late postoperative course of these patients. Moreover, this rare event confirmed that endovascular techniques can play an important role in treating emergent complications. PMID- 16048386 TI - Renal artery thrombosis caused by stent fracture in a single kidney patient. AB - PURPOSE: To report a case of incomplete expansion, fracture, and thrombosis of a stent in the renal artery. CASE REPORT: A 47-year-old man with right renal artery occlusion underwent direct stenting of the left renal artery using a balloon expandable stent. Completion angiography showed satisfactory patency of the vessel although the stent was not fully expanded in its central segment. The patient received 5000 units of heparin during the procedure, but no additional anticoagulant or antiplatelet therapy in the peri/postinterventional period. Twenty-five days later, he presented with acute renal insufficiency and uncontrolled hypertension. Angiography revealed in-stent thrombosis and collateral flow in the distal segment of the left renal artery. He underwent an aortorenal bypass, which salvaged the kidney. The stent, after removal from the vessel, was fractured and not completely expanded. CONCLUSIONS: Incomplete expansion and fracture of the stent associated with insufficient antiplatelet therapy produced in-stent thrombosis. Collateral flow prevented kidney necrosis. PMID- 16048387 TI - Routine bivalirudin use in percutaneous carotid interventions. PMID- 16048388 TI - Stent-grafting of an infected aortoesophageal fistula following ingestion of a fish bone. PMID- 16048389 TI - A comparison of interobserver reproducibility of Gleason grading of prostatic carcinoma in Japan and the United States. AB - CONTEXT: Gleason grading is now the sole prostatic carcinoma grading system recommended by the World Health Organization. It is imperative that there be good interobserver reproducibility within this system worldwide. To our knowledge, there are no studies, using the same specimens, that compare the interobserver reproducibility of Gleason grading in Japan and the United States. OBJECTIVE: To compare the interobserver reproducibility of Gleason grading of prostatic carcinoma in Japan and the United States using, in Japan, images from the identical biopsy glass slides that were originally graded in the United States. DESIGN: Microsopic images from 37 needle biopsies of prostatic carcinoma were placed on CD-ROM and distributed to 14 Japanese pathologists for grading. These 14 physicians included 8 general pathologists and 6 pathologists with a special interest in urologic pathology. The needle biopsies had been previously reviewed so that a consensus diagnosis could be formed by a panel of urologic pathologists in the United States and Canada. Interobserver agreement with the consensus diagnoses was calculated by determining the overall kappa coefficient for the Japanese pathologists and then compared to the interobserver agreement among American general pathologists who had previously graded identical needle biopsies from which the CD-ROM images had been taken. RESULTS: The interobserver agreement with the consensus diagnoses for the 4 Gleason grading groups (Gleason grades 2 4, 5-6, 7, and 8-10) among the Japanese urologic pathologists in this series of cases was substantial (overall kappa = 0.68), and for the Japanese general pathologists, it was moderate (overall kappa = 0.49), similar to that reported in the earlier study of American general pathologists (overall kappa = 0.44). The major interobserver reproducibility problem for both Japanese and American general pathologists is undergrading. The major areas of undergrading are the underdiagnosis of Gleason scores 5-6 as Gleason scores 2-4, and the underdiagnosis of cribriform sheets and fragments of cribriform Gleason pattern 4 carcinoma as Gleason pattern 3. CONCLUSIONS: The interobserver reproducibility of the Gleason grading for this collection of specimens was similar among Japanese and American general pathologists. The overall kappa values for these generalists of 0.44 and 0.49 are only in the moderate (0.41-0.60) range of interobserver agreement when compared to 0.68, substantial (0.61-0.80) agreement, for Japanese urologic pathologists. Educational efforts to improve Gleason grading have been shown to be effective and are clearly warranted. PMID- 16048390 TI - A practical approach to pediatric patients referred with an abnormal coagulation profile. AB - CONTEXT: Workup for prolonged prothrombin time (PT) and activated partial thromboplastin time (PTT) is a frequent referral to a Hematology and Coagulation Laboratory. Although the workup should be performed in a timely and cost effective manner, the complete laboratory assessment of the coagulation state has not been standardized. OBJECTIVE: To determine which clinical and laboratory data are most predictive of a coagulopathy and to formulate the most efficient strategy to reach a diagnosis in patients referred for abnormal coagulation profiles. DESIGN: Retrospective case review. Medical records of 251 patients referred for prolonged PT and/or PTT to our Hematology Service between June 1995 and December 2002 were reviewed. RESULTS: The study included 135 males and 116 females with a mean age of 7.0 years. A personal history of bleeding was reported in 137 patients, and a family history of bleeding was reported in 116 patients. Fifty-one patients (20%) had a coagulopathy (ie, a bleeding risk). Factors predictive of a bleeding risk were a positive family history of bleeding (P < .001) and a positive personal history of bleeding (P = .001). Of 170 patients with findings of normal PT and PTT values on repeat testing, 14 were subsequently diagnosed with a coagulopathy. Two of these patients reported no positive personal or family history of bleeding. CONCLUSIONS: Coagulopathy was identified in 20% of the children referred for abnormal PT and/or PTT. In the absence of a personal or family history of bleeding, a normal PT and/or PTT on repeat testing has a negative predictive value of more than 95%. PMID- 16048391 TI - Distinguishing primary and metastatic conventional renal cell carcinoma from other malignant neoplasms in fine-needle aspiration biopsy specimens. AB - CONTEXT: Differentiation of conventional renal cell carcinoma from other malignant neoplasms can be difficult, particularly with the limited sampling in fine-needle aspiration biopsies. Many studies have discussed the features of renal cell carcinoma in fine-needle aspiration biopsy specimens, but the significance of the cytologic features is not known. OBJECTIVE: To define the significant cytologic findings that aid in differentiating renal cell carcinoma from other malignant neoplasms in fine-needle aspiration biopsy specimens. DESIGN: Fine-needle aspiration biopsies from 35 patients with proven primary or metastatic conventional renal cell carcinoma and from 145 patients with proven primary or metastatic non-renal cell carcinoma malignant neoplasms were assessed for the presence or absence of the following cytologic features: the heterogeneous cell population, hemosiderin deposits, small cytoplasmic vacuoles, large cytoplasmic vacuoles, low nuclear-cytoplasmic ratio, prominent nucleoli, intranuclear inclusions, irregular nuclear membranes, and smooth nuclear membranes. Statistical analysis was performed to identify the features significant in distinguishing conventional renal cell carcinoma from non-renal cell carcinoma malignancies. RESULTS: The presence of the heterogeneous cell population, hemosiderin deposits, small cytoplasmic vacuoles, and low nuclear cytoplasmic ratio were each highly significant in conventional renal cell carcinoma when compared with non-renal cell carcinoma malignant neoplasms (P < .001) using univariate exact analysis. Features that were identified as being predictive of conventional renal cell carcinoma using multivariable logistic regression analysis included heterogeneous cell population, small cytoplasmic vacuoles, and hemosiderin deposits (P < .05). CONCLUSIONS: The presence of the heterogeneous cell population, small cytoplasmic vacuoles, hemosiderin deposits, and a low nuclear-cytoplasmic ratio aids in differentiating conventional renal cell carcinoma from other malignant neoplasms, including morphologically similar entities such as hepatocellular carcinoma, in fine-needle aspiration biopsy specimens. PMID- 16048392 TI - Cutting injuries in an academic pathology department. AB - CONTEXT: Cutting injuries pose an infrequent but serious threat to anatomic pathology personnel. Although cut-resistant gloves may reduce this danger, it is imperative to recognize specific behaviors that increase the chance of an injury. OBJECTIVE: To examine the incidence of cutting injuries in an academic pathology department and the mechanisms by which such injuries occurred. DESIGN: Hospital Report of Event forms completed for laboratory incidents of cutting injury from March 1998 to September 2003 were evaluated. Further information regarding the incidents was obtained, when possible, by interviews with those personnel involved. SETTING: A university-based pathology laboratory was the setting for this study. On average, 505 autopsies and 29,000 surgical specimens were processed each year during the 5.5-year time period. PARTICIPANTS: Pathology attending physicians, residents, dieners, and pathologists' assistants who performed autopsies and surgical specimen examinations. RESULTS: Eight scalpel injuries occurred during the study period. No needle-stick injuries were reported. Searching for lymph nodes and cutting firm tissue each accounted for 3 of the injuries. Only 2 of the 8 individuals were in compliance with the departmental policy regarding protective glove wear. Hospital Report of Event forms alone failed to elicit sufficient detail regarding the mechanism of injury. CONCLUSIONS: A laboratory-based form may be necessary to supplement the hospital form, so as to obtain full details of each injury. This information may then be disseminated to all who handle blades, with the goal of preventing future cutting injuries. PMID- 16048393 TI - Myeloid sarcoma of appendix mimicking acute appendicitis. AB - CONTEXT: Myeloid sarcoma is a neoplasm of immature myeloid cells involving an extramedullary anatomic site that is usually, although not always, associated with acute myeloid leukemia. Any extramedullary site can be involved by myeloid sarcoma, but involvement of the cecal appendix is uncommon, and symptoms mimicking acute appendicitis as a result of appendiceal involvement are rare. OBJECTIVE: To describe the clinicopathologic features of 2 patients with myeloid sarcoma involving the appendix who presented with right lower quadrant pain suggestive of acute appendicitis and prompting appendectomy. DESIGN: Clinical information for both patients was obtained from the medical record. Routine hematoxylin-eosin-stained slides, naphthol-ASD-chloroacetate stain, and immunohistochemical stains for myeloid, B-cell, and T-cell antigens were prepared. RESULTS: Peripheral blood and bone marrow were infiltrated by coexistent acute myeloid leukemia in case 1 but were negative for leukemia in case 2. In case 2, the patient had a history of acute myeloid leukemia that had been treated by an allogenic bone marrow transplant 7 months earlier. Histologic examination of the appendix revealed poorly differentiated myeloid sarcoma in both cases. Each neoplasm was positive for chloroacetate esterase, myeloperoxidase, lysozyme, and CD43 and was negative for CD3 and CD20. CONCLUSIONS: Myeloid sarcoma involving the appendix can rarely cause pain or other symptoms mimicking acute appendicitis. A high index of suspicion combined with the use of cytochemical and immunohistochemical studies are helpful in establishing the diagnosis. PMID- 16048394 TI - Distinguishing pseudoepitheliomatous hyperplasia from squamous cell carcinoma in mucosal biopsy specimens from the head and neck. AB - CONTEXT: The differentiation of pseudoepitheliomatous hyperplasia from invasive squamous cell carcinoma is a difficult and frequently encountered distinction, especially in biopsy specimens from head and neck mucosa. The problem is compounded by inflamed and often poorly oriented tissue sections. OBJECTIVE: To distinguish pseudoepitheliomatous hyperplasia from invasive squamous cell carcinoma, utilizing a panel of antibodies to various epithelial and stromal elements (p53, matrix metalloproteinase 1, E-cadherin, and collagen IV) that has been shown to be useful in differentiating intestinal adenomas with invasive adenocarcinoma from displaced adenomatous epithelium. DESIGN: Thirty-three archival specimens (16 squamous cell carcinoma [12 with invasion and 4 with microinvasion] and 17 pseudoepitheliomatous hyperplasia) from head and neck mucosal locations were immunostained and examined by the authors. RESULTS: We found increased nuclear staining of the invasive tumor cells with p53. There was decreased staining within invasive tumor nests with E-cadherin. There was highly significant increased staining within tumor cells and adjacent stroma with matrix metalloproteinase 1 (P < .001). The only antibody in our panel that did not show a reliable staining pattern was collagen IV. It appeared fragmented in benign inflamed and malignant areas and therefore was not useful. CONCLUSIONS: p53, matrix metalloproteinase 1, and E-cadherin showed significant staining trends independent of inflammation and suboptimal tissue orientation. Although a properly oriented hematoxylin-eosin-stained section was our gold standard, we found this immunoperoxidase panel useful as a diagnostic adjunct in difficult cases. PMID- 16048395 TI - Sessile serrated polyp mimicry in patients with solitary rectal ulcer syndrome: is there evidence of preneoplastic change? AB - CONTEXT: Solitary rectal ulcer syndrome (SRUS) is associated with erythema and ulceration of the rectal wall. Serrated lesions of the colon are divided into conventional hyperplastic polyps and a new set of lesions that are variably called sessile serrated polyps (SSPs) and sessile serrated adenomas. The SSPs are epithelial proliferative lesions that appear to act as a unique pathway to colorectal carcinogenesis. No association between SRUS and SSPs has been previously reported. OBJECTIVE: To assess a possible association between SRUS and morphologic features that mimic SSPs. DESIGN: Twenty-six patients with SRUS, who presented to our institution between January 1, 1999, and November 14, 2004, were retrospectively reviewed for SSP-type morphologic features by 3 pathologists. Ki 67 and hMLH1 immunohistochemical stains were used. Control tissues included 10 conventional left-sided hyperplastic polyps, 10 right-sided large SSPs, 7 adenocarcinomas with known loss of hMLH1 gene expression, and 4 normal human tonsil tissues. RESULTS: Ten (38%) of 26 SRUS specimens demonstrated histologic features consistent with SSPs. These features included exaggerated serration within the lower crypt compartments, crypt branching, hypermucinous appearance of epithelium, and horizontal extension of crypt bases along the muscularis mucosa. All 10 cases of SRUS had positive basal Ki-67 staining in 10% to 20% of cells. Two (20%) of the 10 cases demonstrated focal superficial loss of hMLH1 mismatch repair gene expression within areas of serrated morphologic features. One hyperplastic polyp superimposed on SRUS showed a reduced number of surface epithelial cells that express hMLH1 protein. CONCLUSIONS: Up to 38% of patients with SRUS have histologic changes that mimic SSPs. More importantly, 20% of these serrated lesions were found to have focal loss of hMLH1 gene expression, indicating a potential of preneoplastic change. This phenomenon may reflect an increased propensity for neoplastic progression in response to repeated trauma and repair process in certain cases of SRUS. PMID- 16048397 TI - Uterine-like mass with features of an extrauterine adenomyoma presenting 22 years after total abdominal hysterectomy-bilateral salpingo-oophorectomy: a case report and review of the literature. AB - Adenomyoma is a benign tumor composed of smooth muscle and benign endometrium. These tumors typically originate within the uterus. An extrauterine adenomyoma is a rare entity. We report a uterine-like mass consistent with an extrauterine adenomyoma presenting 22 years following a total abdominal hysterectomy and bilateral salpingo-oophorectomy. The mass was pear-shaped with uterine-type smooth muscle and a cavity lined by functional endometrial glands and stroma. To our knowledge, only 4 other cases of an extrauterine uterine-like mass are reported in the literature. Three involved the ovary, while one was located adjacent to the broad ligament with normal pelvic organs. Although none of these other uterus-like masses were described as adenomyomas with uterine-like features, the histologic findings are strikingly similar. An understanding of the mullerian system suggests that either an embryologic malformation or a differential multipotentiality existing in the subcoelomic tissues in response to hormonal stimulation results in a supernumerary mullerian structure like a uterus, as observed in this case. The presence of endometrial glands and stroma in the mass confirms that the tissues in this mass are hormonally responsive. It is most likely that this uterine-like mass arose from the tissues of the secondary mullerian system in response to estrogenic stimulation. PMID- 16048398 TI - Myocarditis temporally related to the use of gefitinib (Iressa). AB - Gefitinib is a new drug that was approved by the US Food and Drug Administration in May 2003 for non-small cell cancer of the lung refractory to first- and second line therapy. It is regarded as a rather safe drug with common adverse effects that include nausea, vomiting, diarrhea, rash, acne, and dry skin. However, it was reported in Japan to be associated with interstitial pneumonitis (2%-3% of subjects), presumably as a manifestation of a hypersensitivity reaction. The Food and Drug Administration studied this in US patients and found only a 0.3% occurrence and a slightly less than 0.1% mortality due to interstitial pneumonitis. To our knowledge, there has not been an association with fulminant myocarditis or acute myocarditis. We report the case of a 71-year-old man who died as a result of fulminant myocarditis 1 week after starting to take this new class of agent, gefitinib. On the basis of his medical history and our findings, we feel it necessary to consider hypersensitivity myocarditis related to gefitinib the probable cause of death. PMID- 16048399 TI - Monophasic synovial sarcoma of the liver. AB - We report a hepatic monophasic synovial sarcoma in a 60-year-old woman who presented with right upper quadrant pain subsequent to an intrahepatic bleed from a highly vascular tumor mass. Imaging studies showed a dominant tumor mass in the right hepatic lobe with multiple satellite nodules. A detailed physical examination and radiologic workup failed to reveal a primary tumor elsewhere. A right partial hepatectomy was performed with a preoperative differential diagnosis of angiosarcoma versus hepatocellular carcinoma. The morphologic, immunophenotypic, and cytogenetic findings (t(X;18)(p11.2;icq11.2)) were consistent with a monophasic synovial sarcoma. Postoperative clinical evaluation of the extremities and a positron emission tomographic scan performed 4 weeks after surgery showed no evidence of recurrent or metastatic disease. The patient was started on an aggressive 4-drug chemotherapy regimen, but died 3 months thereafter from widespread metastatic disease. No autopsy was performed. The presence of multiple lesions in the liver certainly suggests the possibility of metastatic disease. It would, however, be very unusual for a synovial sarcoma to present as an occult primary, and the negative radiologic workup 1 month after the partial hepatectomy also argues against this possibility. The clinical presentation, radiographic findings, and subsequent course in this patient was therefore most consistent with a primary monophasic synovial sarcoma of the liver. PMID- 16048400 TI - Primary intraocular T-cell-rich large B-cell lymphoma. AB - We report a primary intraocular T-cell-rich large B-cell lymphoma in a 57-year old woman who underwent 3 diagnostic vitrectomies for a presumed diagnosis of panuveitis. She developed no light perception in the left eye and underwent enucleation. Histopathologic and immunohistochemical studies on the enucleated globe disclosed a primary intraocular large B-cell lymphoma involving the choroid, vitreous, and retina. A large population of T cells was identified among the neoplastic B-cell population. B-cell immunoglobulin gene rearrangement and T cell receptor gene rearrangement studies using the polymerase chain reaction method indicated that a monoclonal immunoglobulin kappa light chain population was present and that the T-cell population was not monoclonal. This case highlights the importance of interpreting cytologic features in vitreous aspirates in the context of the clinical situation. PMID- 16048401 TI - Fine-needle aspiration diagnosis of angiosarcoma of the spleen: a case report and review of the literature. AB - Primary angiosarcoma of the spleen is a very rare neoplasm with a poor prognosis. The definitive diagnosis is usually based on the histologic evaluation of the splenectomy specimen. We describe a case of angiosarcoma diagnosed by fine-needle aspiration cytology prior to splenectomy. A 69-year-old white woman presented with heterogeneous lesions in the spleen during a follow-up computed tomographic scan for a history of liposarcoma of the right buttock. A malignant endothelial neoplasm was diagnosed by fine-needle aspiration cytology using immunocytochemistry, and a splenectomy confirmed the presence of angiosarcoma. To our knowledge, this is the first well-documented and confirmed case of primary angiosarcoma of the spleen diagnosed by fine-needle aspiration cytology. This report emphasizes the value of fine-needle aspiration cytology as an important diagnostic tool in splenic neoplasms. PMID- 16048402 TI - Sarcomatoid renal cell carcinoma with divergent sarcomatoid growth patterns: a case report and review of the literature. AB - A 71-year-old woman who presented with left abdominal pain was found to have a noncalcified renal mass with a perisplenic extension on imaging studies. Histologically, the tumor showed predominantly malignant spindle cells with extensive osteoid and chondroid matrix production. Various growth patterns resembling rhabdomyosarcoma, malignant fibrous histiocytoma, and fibrosarcoma were also observed. Immunohistochemistry showed positive staining of the neoplastic cells for cytokeratin and focally positive staining for CD10 and CD117 (c-Kit). Electron microscopic examination revealed a poorly differentiated neoplasm with both mesenchymal and epithelial features. The tumor was diagnosed as a sarcomatoid renal cell carcinoma with overgrowth of the sarcomatoid component (World Health Organization: renal cell carcinoma, unclassified). To our knowledge, sarcomatoid renal cell carcinoma with such a broad morphologic phenotype in a single case has not been documented. Furthermore, the CD117 expression in a sarcomatoid renal cell carcinoma that was observed in this case merits further investigation. PMID- 16048403 TI - Liver biopsy findings following donor lymphocyte infusion. PMID- 16048404 TI - A 37-year-old man with a pleural mass. PMID- 16048405 TI - A 16-year-old adolescent boy with unilateral cervical lymphadenopathy suspicious for malignancy. PMID- 16048406 TI - A 71-year-old man with a bladder mass. PMID- 16048407 TI - Polyomavirus in a lung transplant patient. PMID- 16048409 TI - Storage and transfusion of infected autologous blood or components: a survey of North American laboratories. AB - CONTEXT: Many patients request that autologous blood or components be collected and available for use during scheduled surgical or invasive medical procedures to avoid exposure to human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV) from allogeneic transfusions. Some patients from whom autologous blood is collected are themselves infected with HIV, HBV, or HCV. However, unlike HIV-, HBV-, or HCV-infected allogeneic blood and components, which must be excluded from the community blood supply, infected autologous blood and components are allowed to be stored in hospitals and transfused back to the patients (autologous donors) from whom the blood was collected. Although the transfusion of HIV-, HBV-, or HCV-infected autologous blood or components does not present a risk to the autologous donor, such a transfusion presents a risk to other patients, considering that at least 1 in every 25,000 transfusions are administered to the wrong individual. OBJECTIVE: To determine if hospital transfusion services store and/or transfuse autologous blood or components infected with HIV, HBV, and/or HCV. DESIGN: An educational enhancement subsection of a College of American Pathologists Proficiency Testing Survey (J-C 2003) assessed transfusion service practices for storing and/or transfusing HIV-, HBV-, and HCV-infected autologous blood and components. SETTING AND PARTICIPANTS: A total of 4251 participants were asked whether they stored and/or transfused autologous blood or components and whether these stored blood products included those that were infected with HIV, HBV, or HCV. RESULTS: Of the 4251 survey respondents, 3561 provided data regarding their autologous blood and component storage and/or transfusion practices. A total of 2988 participants reported that they store and/or transfuse autologous blood or components. A total of 2390 respondents reported that they do not test autologous donations collected in their own institution for evidence of infection with HIV, HBV, or HCV. Most survey participants reported that even if an autologous donation is tested and found to be infected they would still be willing to store and transfuse the blood component, according to which agent was causing the infection: HIV (n = 1867), HBV (n = 2158), or HCV (n = 2233). CONCLUSION: Most North American hospitals do not test autologous blood donations that they collect in their own institution for evidence of infection with HIV, HBV, or HCV, leading to the conclusion that infected autologous blood components are being stored and transfused. Even when autologous donations are tested and found to be infected with HIV, HBV, or HCV, most North American hospitals would be willing to store and/or transfuse the infected autologous blood components. PMID- 16048410 TI - North American pretransfusion testing practices, 2001-2004: results from the College of American Pathologists Interlaboratory Comparison Program survey data, 2001-2004. AB - CONTEXT: Pretransfusion testing of whole blood and red blood cell recipients is regulated by the federal government under the authority of the Clinical Laboratory Improvement Amendments of 1988. Regulated tests include determination of ABO group, Rh D type, antibody detection, antibody identification, and crossmatching. A wide variety of methods and reagents are available for these regulated tests. During 2001-2004, the College of American Pathologists (CAP) Interlaboratory Comparison Program (Proficiency Testing) J-Survey collected data from more than 4000 laboratories regarding their pretransfusion testing practices. Those data are presented in this report. OBJECTIVE: To assess current testing practices for ABO grouping, Rh D typing, antibody detection, and crossmatching in North America. DESIGN: Data collected for the CAP Interlaboratory Comparison Program (Proficiency Testing) J-Survey were analyzed for trends in laboratory testing practice during 2001- 2004. The data were grouped for analysis by peer group (testing method used) for ABO grouping, Rh D typing, antibody detection, and crossmatching and then analyzed. SETTING, PATIENTS, OR OTHER PARTICIPANTS: Subscribers to the CAP Interlaboratory Comparison Program Transfusion Medicine J-Series. RESULTS: The most common testing schemes used in North America during 2001-2004 are as follows: ABO grouping (most laboratories perform tube testing: 97.6% in 2000 and 91.1% in 2004); Rh D typing (most laboratories perform tube testing: 97.7% in 2001 and 91.1% in 2004); antibody detection (most laboratories perform tube testing: 69.7% in 2001 and 55% in 2004, most frequently with the low ionic strength solution anti-human globulin [AHG] method, 48.3% in 2001 and 39.9% in 2004; as of 2004 slightly more laboratories use the gel AHG method [42%] than the low ionic strength solution AHG tube method); crossmatching for alloimmunized patients (most laboratories perform tube testing using a low ionic strength solution AHG method; 55.8% in 2001 and 47.6% in 2004); and crossmatching for nonalloimmunized patients (tube testing using an immediate spin method; 42% in 2001 and 40.4% in 2004). CONCLUSIONS: Most North American laboratories currently favor tube methods when performing ABO grouping, Rh typing, antibody screening, and crossmatching. However, there has been a significant increase in the use of gel-based methods in recent years, especially for antibody detection and crossmatching. Data collection and data analysis of CAP Interlaboratory Comparison Program Survey results allow for assessment of laboratory proficiency and provide insights into current North American practice trends in pretransfusion compatibility testing. PMID- 16048411 TI - Activation status of the JAK/STAT3 pathway in mantle cell lymphoma. AB - CONTEXT: Signal transducer and activator of transcription 3 (STAT3) is oncogenic, and we previously found evidence of constitutive STAT3 activation in a relatively small number of frozen mantle cell lymphoma (MCL) cell tumors. OBJECTIVES: To comprehensively survey the activation and phosphorylation status of STAT3 in MCL and to assess if STAT3 activation in these tumors is due to cytokine stimulation by examining the phosphorylation and activation status of Janus kinase (JAK), the physiologic activator of STAT3. DESIGN: We evaluated 43 formalin-fixed, paraffin embedded MCL tumors using immunohistochemistry and phospho-specific antibodies against STAT3 and JAK. RESULTS: There were 37 small cell and 6 blastoid cases. There was heterogeneous expression of phospho-STAT3 (pSTAT3), with 23 negative cases (53%), 12 weakly positive cases (28%), and 8 strongly positive cases (19%). JAK3 was the only member detectable in 3 MCL cell lines, and immunoprecipitation data showed a relatively low level of tyrosine phosphorylation of JAK3 in these cells. Using immunohistochemistry, phospho-JAK3 (pJAK3) was detectable in 18 (44%) of 41 MCL tumors examined, and pJAK3 expression correlated with that of pSTAT3 (P = .008). A notable exception to this correlation was seen in the blastoid variant, since 4 (67%) of 6 blastoid cases were pSTAT3 positive but pJAK3 negative. CONCLUSIONS: We have confirmed our previous finding that STAT3 is constitutively activated in MCL tumors, with an overall frequency of 47% in this series. STAT3 activation in the small cell but not the blastoid variant of MCL is likely mediated by JAK3. PMID- 16048412 TI - Performance characteristics of several rules for self-interpretation of proficiency testing data. AB - CONTEXT: Proficiency testing (PT) participants can interpret their results to detect errors even when their performance is acceptable according to the limits set by the PT provider. OBJECTIVE: To determine which rules for interpreting PT data provide optimal performance for PT with 5 samples per event. DESIGN: We used Monte Carlo computer simulation techniques to study the performance of several rules, relating their error detection capabilities to (1) the analytic quality of the method, (2) the probability of failing PT, and (3) the ratio of the peer group SD to the mean intralaboratory SD. Analytic quality is indicated by the ratio of the PT allowable error to the intralaboratory SD. Failure of PT was defined (Clinical Laboratory Improvement Amendments of 1988) as an event when 2 or more results out of 5 exceeded acceptable limits. We investigated rules with limits based on the SD index, the mean SD index, and percentages of allowable error. RESULTS: No single rule performs optimally across the range of method quality. CONCLUSIONS: We recommend further investigation when PT data cause rejection by any of the following 3 rules: any result exceeds 75% of allowable error, the difference between any 2 results exceeds 4 times the peer group SD, or the mean SD index of all 5 results exceeds 1.5. As method quality increases from marginal to high, false rejections range from 16% to nearly zero, and the probability of detecting a shift equal to 2 times the intralaboratory SD ranges from 94% to 69%. PMID- 16048413 TI - A 14-year-old girl with a retroperitoneal soft tissue mass. PMID- 16048414 TI - A 46-year-old man with an enlarging finger nodule. PMID- 16048415 TI - A 17-month-old girl with erythrocytosis. PMID- 16048416 TI - Fatal drug reaction due to allopurinol therapy in a 72-year-old man. PMID- 16048417 TI - A 27-year-old woman with an incidental renal mass. PMID- 16048418 TI - Duodenal polyp in a 75-year-old man. PMID- 16048419 TI - A future with increased rather than diminished research capabilities. PMID- 16048420 TI - Sensitivity to change and responsiveness of the global physiotherapy examination (GPE-52) in patients with long-lasting musculoskeletal pain. AB - BACKGROUND AND PURPOSE: The sensitivity to change and the responsiveness of the Global Physiotherapy Examination (GPE-52) were examined in patients with localized and widespread long-lasting musculoskeletal pain. SUBJECTS: All included patients were on sick leave and constituted the treatment group in a randomized controlled study. Two hundred forty-seven patients (64% women; mean age = 43.6 years, SD = 10.4) were included. At entry, the patients were categorized into 3 groups according to pain localization. Patients who returned fully to work were compared with those who did not. METHODS: The GPE-52 has composite scores in 5 main domains--posture (8 tests), respiration (8 tests), movement (16 tests), muscle (12 tests), and skin (8 tests)--and was administered before and after 4 weeks of multidisciplinary outpatient treatment and at a 6 month follow-up examination. The first 61 patients also were re-examined after 18 months. RESULTS: In this study, the GPE-52 and its 5 main domains were demonstrated to be sensitive to change, measured 6 and 18 months after treatment, in different groups of patients with long-lasting musculoskeletal pain. Responsiveness to important change, defined in this study as return to work, was found only for the total GPE-52 score and within the movement and respiration domains. Responsiveness to important change was greater in patients with localized pain than in patients with widespread pain. DISCUSSION AND CONCLUSION: Although the total GPE-52 score was sensitive to change in patients with long lasting musculoskeletal pain, a shorter test battery including only the respiration and movement domain scores might be appropriate as an outcome measure in intervention studies for patients with long-lasting musculoskeletal pain. PMID- 16048421 TI - Can elderly patients who have had a hip fracture perform moderate- to high intensity exercise at home? AB - BACKGROUND AND PURPOSE: The majority of patients after a hip fracture do not return to prefracture functional status. Depression has been shown to affect recovery. Although exercise can reduce impairments, access issues limit elderly people from participating in facility-based programs. The primary purpose of this study was to determine the effects and feasibility of a home exercise program of moderate- or high-intensity exercise. A secondary purpose was to explore the relationship of depression and physical recovery. SUBJECTS: Thirty-three elderly people (24 women, 9 men; mean = 78.6 years of age, SD = 6.8, range = 64-89) who had completed a regimen of physical therapy following hip fracture participated in the study. Subjects were randomly assigned to a resistance training group, an aerobic training group, or a control group. METHODS: Subjects were tested before and upon completion of the exercise trial. Isometric lower-extremity force, 6 minute-walk distance, free gait speed, mental status, and physical function were measured. Each exercise session was supervised by a physical therapist, and subjects received 20 visits over 12 weeks. The control group received biweekly mailings. The resistance training group performed 3 sets of 8 repetitions at the 8-repetition maximum intensity using a portable progressive resistance exercise machine. The aerobic training group performed activities that increased heart rate 65% to 75% of their age-predicted maximum for 20 continuous minutes. RESULTS: Resistance and aerobic training were performed without apparent adverse effects, and adherence was 98%. All groups improved in distance walked, force produced, gait speed, and physical function. Isometric force improved to a greater extent in the intervention groups than in the control group. Depressive symptoms interacted with treatment group in explaining the outcomes of 6-minute walk distance and gait speed. DISCUSSION AND CONCLUSION: High-intensity exercise performed in the home is feasible for people with hip fracture. Larger sample sizes may be necessary to determine whether the exercise regimen is effective in reducing impairments and improving function. Depression may play a role in the level of improvement attained. PMID- 16048422 TI - Perturbation training improves knee kinematics and reduces muscle co-contraction after complete unilateral anterior cruciate ligament rupture. AB - BACKGROUND AND PURPOSE: Dynamic knee stabilization strategies of people who successfully compensate for the absence of an anterior cruciate ligament (ACL) ("copers") are different from those of people who do not compensate well for the injury ("noncopers"). Early after injury, certain patients ("potential copers") can increase the likelihood of successfully compensating for the injury by participating in 10 sessions of perturbation training. The purpose of this study was to determine how perturbation training alters muscle co-contraction and knee kinematics in potential copers. SUBJECTS: Seventeen individuals with acute, unilateral ACL rupture who were categorized as potential copers and 17 subjects without injuries who were matched by age, sex, and activity level were recruited for this study. METHODS: Motion analysis and electromyographic data were collected as subjects walked across a stationary or moving platform (horizontal translation) before and after perturbation training. RESULTS: Before training, potential copers had higher co-contraction indexes and lower peak knee flexion angles than subjects without injuries. After training, potential copers' movement patterns more closely resembled those of subjects without injuries (ie, they showed reduced co-contraction indexes and increased peak knee flexion angles during stance). DISCUSSION AND CONCLUSION: Perturbation training reduced quadriceps femoris-hamstring muscle and quadriceps femoris-gastrocnemius muscle co-contractions and normalized knee kinematics in individuals with ACL rupture who were classified as potential copers. Findings from this study provide evidence for a mechanism by which perturbation training acts as an effective intervention for promoting coordinated muscle activity in a select population of people with ACL rupture. PMID- 16048423 TI - Electromyographic analyses of global synkinesis in the paretic upper limb after stroke. AB - BACKGROUND AND PURPOSE: Global synkinesis (GS), or motor irradiation, is an involuntary movement associated with the coactivation of numerous muscles in one limb when the opposite limb is active. The electromyographic (EMG) patterns of people with stroke and people who were healthy were analyzed to characterize GS development in relation to joint involvement and to attempt to relate these findings to clinical observations. SUBJECTS AND METHODS: Twenty patients with stroke, divided into 2 groups with either greater levels of irradiation (SG, n = 10) or lesser levels of irradiation (SL, n = 10), and 20 subjects in a control group were studied. A dynamometer was used to provide resistance for voluntary isometric muscle contractions of the flexor muscle groups of the shoulder, elbow, and wrist. The summated and standardized net EMG amplitudes of 8 principal muscles of the unexercised (paretic) upper extremity were used to characterize intensity and spatial representation of GS. Clinical measurements included the Fugl-Meyer Assessment Scale (FMA), Barthel Index of Activities of Daily Living (BI), and the stage on the Brunnstrom Stages of Motor Recovery Scale (BR). RESULTS: In the SG and control groups, a more substantial GS intensity was associated with muscle contractions of the flexor muscles of the opposite proximal joint than was the case for contractions of the flexor muscles of the distal joint, whereas such a gradient change was absent in the SL group. The corresponding spatial patterns of GS exhibited a predominant cross-excitation over the unexercised pectoralis major and extensor carpi radialis muscles in the control group, contrary to the enhanced activation of the brachioradialis and biceps brachii muscles noted in patients with stroke. The SG group had a better FMA score and a more satisfactory BR stage than did the SL group, and the 2 neurological scores were related to GS intensity for patients with stroke, depending on joint involvement. DISCUSSION AND CONCLUSION: Intensity of GS provided an affiliation with motor deficits and a promising window for poststroke recovery mechanisms. PMID- 16048424 TI - Association of importance of the doctoral degree with students' perceptions and anticipated activities reflecting professionalism. AB - BACKGROUND AND PURPOSE: The American Physical Therapy Association (APTA) has identified the Doctor of Physical Therapy (DPT) degree as 1 of 6 elements necessary to transition the physical therapy profession to a fully professionalized discipline. However, there have been no data to determine whether physical therapist students who place importance on the DPT degree perceive physical therapy to be more professionalized or anticipate participation in activities reflecting professionalism more than those who do not place importance on the DPT degree. SUBJECTS: The subjects were 919 professional physical therapist students. METHODS: Faculty members at 34 physical therapist education programs distributed questionnaires to 1,172 professional physical therapist students and returned 919 questionnaires, for a response rate of 78.4%. The data were statistically analyzed using chi-square analysis and logistic regression. RESULTS: There were few differences between students who place importance on the DPT degree (DPT-I students) and those who do not place importance on the DPT degree (DPT-NI students) regarding how professionalized they perceive physical therapy to be relative to other health care professions or regarding their anticipated participation in activities reflecting professionalism. The one potential distinction found when controlling for other variables was that DPT-I students were more likely than DPT-NI students to anticipate becoming faculty members. DISCUSSION AND CONCLUSION: When beginning their professional education, there are few differences between DPT-I and DPT-NI students' perceptions of the professionalization of physical therapy or anticipation of activities reflecting professionalism. PMID- 16048425 TI - Proficiency of balance in children and youth who have had acute lymphoblastic leukemia. AB - BACKGROUND AND PURPOSE: As the survival rate for acute lymphoblastic leukemia (ALL) in childhood increases, long-term sequelae are a growing concern. This cross-sectional, descriptive study compared the balance skills of children and youth who have had ALL with those of comparable subjects and explored associations with demographics, therapy, physical activity, and health-related quality of life (HRQL). SUBJECTS: Ninety-nine subjects treated previously for ALL and 89 comparison subjects were examined. METHODS: Measures included the Bruininks-Oseretsky Test of Motor Proficiency (BOTMP) balance subtest, the Children's Self-perceptions of Adequacy in and Predilection for Physical Activity Scale (CSAPPA), and the Health Utilities Index (HUI), a measure of HRQL. RESULTS: The children and youth who had ALL had poorer balance than the comparison subjects (BOTMP = 10.55 and 16.30, respectively) and lower CSAPPA scores (57.72 and 63.72, respectively) and HUI scores (0.86 and 0.97, respectively). Regression analyses identified exposure to cranial irradiation, being overweight, lower CSAPPA scores for adequacy, and lower HUI single-attribute scores for cognition as predictors of lower balance scores in subjects who had ALL. DISCUSSION AND CONCLUSION: Balance abilities in subjects treated for ALL were compromised, and several factors were associated with this deficit. PMID- 16048426 TI - Specialized neuromuscular training to improve neuromuscular function and biomechanics in a patient with quiescent juvenile rheumatoid arthritis. AB - BACKGROUND AND PURPOSE: The purpose of this case report is to describe a novel multidisciplinary approach for evaluating and preparing a patient with quiescent juvenile rheumatoid arthritis (JRA) for safe sports participation. CASE DESCRIPTION: The patient was a 10-year-old girl with a history of bilateral knee arthritis who desired to participate in soccer and basketball. Range of motion and manual muscle testing of the lower extremity were within normal limits. Neuromuscular testing included kinematic and kinetic testing, isokinetic assessment, and postural stability testing. The patient's gait was near normal; however, she had narrowed step width and increased knee flexion at heel-strike. Landing analysis during a box drop vertical jump task showed increased and imbalanced (right versus left lower extremity) peak impact forces. The testing was followed by specialized neuromuscular training (SNT). OUTCOMES: Following SNT, heel-strike and step width were within normal limits, peak impact forces on the box drop test decreased by 31%, imbalance decreased by 46%, and vertical jump increased 15%. The isokinetic strength ratio between knee flexors and extensors and the overall balance measures were within normal limits and equal bilaterally. DISCUSSION: Patients with quiescent JRA may have abnormal biomechanics, which could place them at increased risk for injury or future articular cartilage damage. Specialized neuromuscular training may have helped to decrease the patient's risk for future injury or disease progression. PMID- 16048427 TI - [Budd-Chiari sindrome and hererozygotic mutation of metylenetetrahidrofolate reductase.] PMID- 16048428 TI - [Sedation in gastrointestinal endoscopy]. PMID- 16048429 TI - [Obstructive jaundice in a 22- year-old man]. PMID- 16048430 TI - [Hepatic fibrosis after antidepressant treatment]. PMID- 16048431 TI - [Emetic syndrome of uncommon origin]. PMID- 16048432 TI - Defibrillation beliefs of rural nurses: focus group discussions guided by the Theory of Planned Behaviour. AB - INTRODUCTION: The endorsement of the chain of survival concept and early defibrillation has challenged health professionals to reconsider their beliefs about how they respond to in-hospital resuscitation. In the rural context, where 24 hour coverage is not available nurse-initiated defibrillation is expected. Despite literature and policy change in Australia to allow nurses to initiate defibrillation, there is no current research that uses a systemic theoretical approach to investigate the specific beliefs of nurses and their use of defibrillators. The purpose of this study was to elicit a beginning understanding of the defibrillation beliefs of rural nurses. METHODS: This research used focus groups within the framework of the Theory of Planned Behavior to describe the defibrillation beliefs of rural registered nurses. The sites selected for this study were two acute care hospitals in rural Australia (RRMA Classification). Each of these hospitals was in located 'other rural areas' (RRMA Classification) in separate towns and had 25 and 30 beds. The study sample consisted of 10 females and two males. Focus group questions were designed to elicit salient beliefs within the theoretical framework. Three constructs of behavioral, normative and control beliefs guided the development of the question and analysis of the discussions. In accordance with the authors of the theoretical framework, content analysis was used to analyse the data from the study. RESULTS: Two behavioral beliefs, four control beliefs and four normative belief categories were elicited. Two behavioral beliefs categories emerged from the open-ended question: 'What, if any are the advantages of you being able to use a defibrillator?' Participants were congruent when discussing the advantages of nurses initiating defibrillation. The two categories were 'quicker response times' (15 responses) and 'increased success with resuscitation' (8 responses). Participants were asked to identify any events that might influence their decision to use or not use a defibrillator if there was a cardiac arrest on their ward on that day. The categories of control beliefs elicited were 'rhythm recognition' (22 responses), 'litigation' (15 responses), 'fear of harm to patient or self' (11 responses), and 'roles' (4 responses). To identify the normative referents, participants were asked to identify who would approve or not approve of them being responsible for the use of defibrillators in their clinical area. Four normative beliefs represent 100% of the responses, these were: patients; nurses; doctors; and the nursing registration body, the Queensland Nursing Council. CONCLUSIONS: The central issues for these participating nurses were related to the consequences for the patient, support and confidence with rhythm recognition. Understanding rural nurses beliefs as they pertain to nurse initiated defibrillation may provide educators with some insight as to what changes are needed to increase nurse-initiated defibrillation. PMID- 16048434 TI - The granule pathway of programmed cell death. AB - The exocytosis of death-inducing granzymes stored in the granules of cytotoxic lymphocytes allows the immune system to rapidly eliminate intracellular pathogens and transformed cells. The membrane-disrupting protein perforin allows the entry of granzymes into a cell, where they induce apoptosis by cleaving target substrates in the cytoplasm and nucleus. Granzymes kill cells in a variety of ways. Recent work has demonstrated that granzymes induce mitochondrial dysfunction through caspase and caspase-independent pathways and destroy DNA and the integrity of the nucleus. Cytotoxic lymphocytes are susceptible to self inflicted damage. Mice and humans defective in perforin and granzymes point to a role for self-inflicted damage in downregulating lymphocyte responses. Given the propensity for the granule pathway to inflict cellular damage, cytotoxic lymphocytes have developed a variety of mechanisms to protect themselves. In this regard, endogenous serine protease inhibitors have been suggested to protect cytotoxic lymphocytes from granzyme B. It would appear that certain viruses and possibly even tumor cells also use the same mechanism to escape destruction from the exocytosis pathway of programmed cell death. PMID- 16048435 TI - iNKT-cell responses to glycolipids. AB - Invariant natural killer T (iNKT) cells are an unusual group of T lymphocytes that recognize glycolipid antigens presented by the major histocompatibility complex class I-related protein CD1d. Because iNKT cells play a regulatory role in the immune system, they are attractive targets for immunotherapy. The marine sponge-derived glycolipid alpha-galactosylceramide (alpha-GalCer) potently activates iNKT cells. In vivo administration of alpha-GalCer to mice or humans results in rapid and robust cytokine secretion by iNKT cells, followed by the activation of a variety of cell types of the innate and adaptive immune systems. These potent immunomodulatory activities of alpha-GalCer are being exploited for therapeutic purposes. Preclinical studies in mice have demonstrated that alpha GalCer and related glycolipids can protect mice against a variety of diseases, including cancer, infections, and several autoimmune and inflammatory conditions. Although alpha-GalCer treatment of mice is associated with unwanted side-effects, it has been proven safe in clinical trials with cancer patients. These studies have raised significant enthusiasm for the development of effective and safe iNKT cell-based immunotherapies for a variety of human diseases. PMID- 16048436 TI - Dendritic cell-derived exosomes as cell-free peptide-based vaccines. AB - Dendritic cells (DC) are professional antigen-presenting cells and the only ones capable of inducing primary cytotoxic immune responses both in vivo and in vitro. DCs secrete a 60-100 nm membrane vesicle population of endocytic origin, called exosomes. The lipid and protein composition of DC-derived exosomes (DEX) is now well characterized. Besides MHC and costimulatory molecules, DEX bear several adhesion proteins, which are probably involved in their specific targeting. DEX also accumulate several cytosolic factors, most likely involved in exosome's biogenesis in late endosomes. In 1998, we reported that DEX are immunogenic in mice and lead to tumor rejection. These findings have renewed the interest in DEX. The current challenge consists of understanding the mechanisms and the physiological relevance of DEX, which could contribute to the design of the optimal DEX-based vaccination. In this review, we focus on the biological features of DEX and their immunostimulatory functions in mice and humans, and we discuss their potential clinical implementation in the immunotherapy of cancer. PMID- 16048437 TI - How can the innate immune system influence autoimmunity in type 1 diabetes and other autoimmune disorders? AB - The environment is important in determining the onset of autoimmune diseases such as type 1 diabetes. Genetic susceptibility factors interact with the environment to trigger and modulate a series of immune responses that ultimately lead to destruction of the insulin-producing beta cells of the pancreas. Although T lymphocytes are thought to play a major pathogenic role in the pathogenesis of diabetes, undoubtedly other components of the immune system also contribute to this process. How the environment may alter the course of disease is unknown, although viruses have been implicated in triggering and/or exacerbating the disease process. In this review, we will focus on how infection, particularly with viruses, may influence the onset of type 1 diabetes and other autoimmune diseases. Mechanisms such as molecular mimicry, bystander activation, and uncontrolled polyclonal activation of the immune system may contribute to the immune pathogenesis. We will also explore the interaction of the innate immune system with adaptive immune responses in predisposing individuals to the development of autoimmunity. PMID- 16048438 TI - STAT3-mediated transcription of Bcl-2, Mcl-1 and c-IAP2 prevents apoptosis in polyamine-depleted cells. AB - Activation of STAT3 (signal transducer and activator of transcription 3) plays a crucial role in cell survival and proliferation. The aim of the present study was to clarify the role of STAT3 signalling in the protection of polyamine-depleted intestinal epithelial cells against TNF-alpha (tumour necrosis factor-alpha) induced apoptosis. Polyamine depletion by DFMO (alpha-difluoromethylornithine) caused phosphorylation of STAT3 at Tyr-705 and Ser-727. Phospho-Tyr-705 STAT3 was immunolocalized at the cell periphery and nucleus, whereas phospho-Ser-727 STAT3 was predominantly detected in the nucleus of polyamine-depleted cells. Sustained phosphorylation of STAT3 at tyrosine residues was observed in polyamine-depleted cells after exposure to TNF-alpha. Inhibition of STAT3 activation by AG490 or cell-membrane-permeant inhibitory peptide (PpYLKTK; where pY represents phospho Tyr) increased the sensitivity of polyamine-depleted cells to apoptosis. Expression of DN-STAT3 (dominant negative-STAT3) completely eliminated the protective effect of DFMO against TNF-alpha-induced apoptosis. Polyamine depletion increased mRNA and protein levels for Bcl-2, Mcl-1 (myeloid cell leukaemia-1) and c-IAP2 (inhibitor of apoptosis protein-2). Significantly higher levels of Bcl-2 and c-IAP2 proteins were observed in polyamine-depleted cells before and after 9 h of TNF-alpha treatment. Inhibition of STAT3 by AG490 and DN STAT3 decreased Bcl-2 promoter activity. DN-STAT3 decreased mRNA and protein levels for Bcl-2, Mcl-1 and c-IAP2 in polyamine-depleted cells. siRNA (small interfering RNA)-mediated inhibition of Bcl-2, Mcl-1 and c-IAP2 protein levels increased TNF-alpha-induced apoptosis. DN-STAT3 induced the activation of caspase 3 and PARP [poly(ADP-ribose) polymerase] cleavage in polyamine-depleted cells. These results suggest that activation of STAT3 in response to polyamine depletion increases the transcription and subsequent expression of anti-apoptotic Bcl-2 and IAP family proteins and thereby promotes survival of cells against TNF-alpha induced apoptosis. PMID- 16048439 TI - Bcl-xL inhibits T-cell apoptosis induced by expression of SARS coronavirus E protein in the absence of growth factors. AB - One of the hallmark findings in patients suffering from SARS (severe acute respiratory syndrome) is lymphopenia, which is the result of massive lymphocyte death. SARS-CoV (SARS coronavirus), a novel coronavirus that has been etiologically associated with SARS cases, is homologous with MHV (murine hepatitis coronavirus), and MHV small envelope E protein is capable of inducing apoptosis. We hypothesized that SARS-CoV encodes a small envelope E protein that is homologous with MHV E protein, thus inducing T-cell apoptosis. To test this hypothesis, a cDNA encoding SARS-CoV E protein was created using whole gene synthesis. Our results showed that SARS-CoV E protein induced apoptosis in the transfected Jurkat T-cells, which was amplified to higher apoptosis rates in the absence of growth factors. However, apoptosis was inhibited by overexpressed antiapoptotic protein Bcl-xL. Moreover, we found that SARS-CoV E protein interacted with Bcl-xL in vitro and endogenous Bcl-xL in vivo and that Bcl-xL interaction with SARS-CoV E protein was mediated by BH3 (Bcl-2 homology domain 3) of Bcl-xL. Finally, we identified a novel BH3-like region located in the C terminal cytosolic domain of SARS-CoV E protein, which mediates its binding to Bcl-xL. These results demonstrate, for the first time, a novel molecular mechanism of T-cell apoptosis that contributes to the SARS-CoV-induced lymphopenia observed in most SARS patients. PMID- 16048440 TI - Suicidal ideation and associated factors among community-dwelling elders in Taiwan. AB - The purpose of the present study was to explore the suicidal ideation of community-dwelling elderly and the factors associated with their intention to commit suicide. Using a multilevel stratified sampling strategy, 1000 elderly subjects were recruited (aged 65-74 years old) in Taiwan during the year 2001. The degree of depression and its correlates were assessed. Suicidal ideation was measured by asking respondents if they had had any suicidal thoughts in the previous week. In all, 16.7% of respondents reported suicidal ideation within the past week; its occurrence was related to sex, religious belief, employment status, marital status, average family monthly income, physical health status, depressive symptoms, and community activity participation. Further multivariate logistic regression revealed that, aside from depressive symptoms and a lower level of education, no community participation in the past 6 months was significantly associated with the appearance of suicidal ideation. The prevalence of suicidal ideation among the elderly in Taiwan is higher than in Western countries. Participation in social activities is negatively associated with elderly suicidal ideation. The dimension of social participation deserves further exploration and should be considered in community mental health promotion interventions for elderly people. PMID- 16048441 TI - Influence of depression and HIV serostatus on the neuropsychological performance of injecting drug users. AB - Depression is common in injecting drug users (IDUs), a group at significant risk for HIV infection. Moreover, both HIV infection and depression have been shown to adversely effect neurocognitive abilities. Understanding the effects of depression and HIV infection on the neurocognitive functioning of drug users is essential for appropriate management and/or treatment of these deficits in this population. Therefore, the purpose of the present study was to investigate the effects of depression and HIV status on cognitive functioning in 100 male and female IDUs. Participants were categorized into three groups of depression severity based on their scores on the Beck Depression Inventory: no depression, mild depression, and moderate to severe depression. The effects of depression and HIV serostatus as well as their interaction were assessed. Results indicated that regardless of serostatus, those with moderate to severe depression had lower scores on cognitive measures. These findings suggest that although depression contributes to poor neuropsychological performance in IDUs, this effect was not exacerbated by HIV infection. The finding also illustrates the importance of addressing depression-related neurocognitive deficits in IDUs. PMID- 16048442 TI - Interception of potential adverse drug events in long-term psychiatric care units. AB - The interception of medication errors is required for patient safety. The aim of the present study was to clarify factors associated with the interception of potential adverse drug events in long-term psychiatric care units. A survey was conducted of medication-related errors in 132 Japanese long-term psychiatric care units for 2 months using an incident reporting system. The relationship was analyzed between the reported potential adverse drug events and the characteristics of the units and the staff, as well as those of the patients involved. A multivariate logistic regression analysis was performed with environmental, organizational, and human factors as independent variables to predict the interception of potential adverse drug events. Of the 221 reported incidents, 55 (24.9%) were intercepted before reaching patients. The following patient groups were significantly associated with the failure to intercept potential adverse drug events: patients receiving a relatively large number of tablets, patients with relatively frequent admissions, and patients exposed to a relatively high patient-staff ratio in the evening. In contrast, patients with a diagnosis of schizophrenia were significantly associated with an increased possibility of interception. To intercept more potential adverse drug events, simpler prescriptions are crucial. To improve the current situation, organizational efforts, such as increasing staff in the evening, and educating the staff about medications, will be required, as well as improvements in the medications themselves. PMID- 16048443 TI - Pharmacoeconomic evaluation of schizophrenia in Taiwan: model comparison of long acting risperidone versus olanzapine versus depot haloperidol based on estimated costs. AB - Antipsychotics are the keystone in schizophrenia treatment. Although the benefits of the new generation of antipsychotics has been demonstrated over the last decade, the issues of patient compliance and higher purchasing price of atypical antipsychotics remain unresolved. Risperidone is the only atypical antipsychotic agent with long-acting formulation. Long-acting risperidone is a water-based injection and it has been associated with a low level of pain. The aim of the present study was to test whether an improvement in compliance with the use of a long-acting risperidone, compared with olanzapine and depot haloperidol, can increase the effectiveness and the cost-effectiveness indexes. An economic comparison model with decision tree, rather than a prospective design with real clinical drug trial, was applied. The unit cost for each medical procedure was obtained from the claimed-database of the Bureau of National Health Insurance in Taiwan. An executive committee simulated the incidence of extrapyramidal side effects and proposed a therapeutic model for each strategy based on a literature review. The probabilities of treatment response of different agents and those of different mental health states were estimated by the executive committee and 10 senior psychiatrists who were randomly selected. Sensitivity analysis was performed for drug cost-effectiveness and compliance improvement for using long acting risperidone. The results showed that long-acting risperidone is more cost effective than either olanzapine or depot haloperidol for treating schizophrenia patients whose conditions are stable and whose illness duration ranges from 1 to 5 years. The comparison model with the Kaplan-Meier decision tree may serve as an alternative to prospectively designed studies for cost-effectiveness of atypical antipsychotics. PMID- 16048444 TI - Prevalence of insomnia and its relationship to menopausal status in middle-aged Korean women. AB - Although the prevalence of insomnia and the association of insomnia with menopause have been well reported, not much work has been conducted in population based research on insomnia and menopause in Korea. The purpose of the present report was to determine overall and different prevalence of insomnia by menopausal status, and the relationship between insomnia and menopause in a population-based sample of middle-aged Korean women. A total of 96.1% of 2497 randomly selected middle-aged Korean women participated. Insomnia was defined as occurring three times a week or more in the previous month. Subjects were categorized into three groups: premenopause, perimenopause, and postmenopause. The overall prevalence of insomnia in middle-aged Korean women was 14.3%. The most common symptom of insomnia was difficulty maintaining sleep (9.7%), followed by difficulty initiating sleep (7.9%), and early morning awakening (7.5%). Multiple logistic regression analysis revealed that menopause was independently associated with insomnia after adjusting for confounding factors such as age, income, and depression. Perimenopause was significantly associated with a dramatic increase in the risk of insomnia, but there was no significant association for postmenopause. The major finding is that insomnia is significantly associated with the menopausal transition. The prevalence of insomnia increases significantly by the transition from premenopause to perimenopause, but not to postmenopause. A further prospective study is needed to investigate the influence of menopause on insomnia. PMID- 16048445 TI - Relationships between insight and medication adherence in outpatients with schizophrenia and bipolar disorder: prospective study. AB - The aim of the present study was to explore the associations between insight and medication adherence at index interview and at 1-year follow-up interview in bipolar and schizophrenic outpatients. The Schedule for Assessment of Insight (SAI) and its expanded version (SAI-E) were used to provide a baseline insight score for 65 bipolar subjects and 74 schizophrenic subjects considered to be in remission or to have minimal psychopathology. Medication adherence of subjects was assessed at index interview and at 1-year follow-up interview, and the association between insight and medication adherence was analyzed cross sectionally and prospectively. The results of the analysis reveal that in bipolar subjects, the index SAI scores for insight into treatment, mental-health status and psychotic experiences, and total SAI-E were positively correlated with medication adherence at both index and 1-year follow-up interviews. However, in schizophrenic subjects, insight into treatment and total SAI-E correlated positively with medication adherence at index interview but not at 1-year follow up interview. Medication adherence at index interview could predict medication adherence 1 year later in both bipolar and schizophrenic subjects. These results indicate that the predictive value of insight for medication adherence differs between bipolar and schizophrenic patients, and building insight is an important step for establishing medication adherence in bipolar patients. PMID- 16048446 TI - Tripartite relationship among P300, clinical features and brain structure in neuroleptic-naive schizophrenia. AB - Auditory P300 abnormalities in schizophrenia patients have been repeatedly reported by many studies. However, reported relationships among P300 abnormalities, clinical features and other biological variables, such as abnormalities in structural brain imaging, are notably discrepant. This is partially due to the inclusion of patients who have had long-term administration of neuroleptics and those from whom this treatment has been withdrawn. The present study measures event-related potentials in 13 neuroleptic-naive schizophrenia patients using an auditory oddball paradigm to clarify the relationships among P300 amplitude, clinical features and brain structure. All patients underwent computed tomography to estimate the area of the right and left frontal cortical sulci and Sylvian fissures. Clinical symptoms were assessed using the Positive And Negative Syndrome Scale. The high correlation coefficients were obtained between P300 amplitude and the anxiety/depression factor score (r = -0.77), the positive factor score (r = -0.58) and between P300 amplitude and the area ratios of the fronto-temporal region (r = -0.66). These findings show that fronto-temporal region and P300 amplitude are closely related to the earliest stage of illness even in neuroleptic-naive patients. PMID- 16048447 TI - Latency of auditory P300 correlates with self-control as measured by the Sixteen Personality Factor Questionnaire. AB - The reception, processing, and storage of information about experience define personality. The present study investigated the relationship between auditory event-related potentials (AERP) and personality traits. The AERP were recorded using a standard auditory oddball paradigm, and personality was evaluated by Cattell's Sixteen Personality Factor Questionnaire (16PF) in 20 healthy young male subjects. The P300 latency was found to be significantly associated with rule consciousness (factor G in the 16PF), perfectionism (factor Q3), and self control (factor SC): it was negatively correlated with G score (r = -0.56, P = 0.01), Q3 score (r = -0.67, P = 0.001), and SC score (r = -0.65, P = 0.002). Moreover, the P300 amplitude and N100 amplitude were negatively correlated with reasoning (factor B; r = -0.46, P = 0.044; and r = -0.72, P = 0.002, respectively). These results indicate that the personality traits of self control, perfectionism, high superego, and reasoning are related to information processing in the brain. PMID- 16048448 TI - Clinical application of single-pulse transcranial magnetic stimulation for the treatment of depression. AB - Transcranial magnetic stimulation (TMS) has been recently suggested for the treatment of patients with major depression. Based on the results of the authors' pilot study showing a possible antidepressive effect of single-pulse TMS, a clinical trial was conducted involving patients with major depression. For the present study single-photon emission computed tomography (SPECT) was recorded for six of the target patients to study the effects of TMS on the local blood flow volume. Twenty-three inpatients meeting the Diagnostic and Statistical Manual of Mental Disorders (4th edn; DSM-IV) criteria for major depression were invited to participate in the study. Depressive symptoms were rated using the Hamilton Rating Scale for Depression (HAM-D). Patients were given 10 stimuli over the frontal area of both sides for a total of 20 stimuli in a session. The subjects had daily TMS session for 5 days as an add-on therapy. In addition, six patients had their quantitative (99m)Tc-ethyl cysteinate dimer SPECT images measured before and after TMS treatment. Compared with the value 2 days prior to the start of TMS therapy (24.2 +/- 4.9), the average HAM-D scale dropped significantly to 15.3 +/- 6.6 on the day after completion of such therapy. The results of SPECT showed that the regional cerebral blood flow (rCBF) of the bilateral frontal region had increased in four out of six patients when comparing before and after treatment. The present study shows that single-pulse TMS, which is widely used as a neurological test method, possesses a wide range of antidepressive effects without inducing adverse reactions. The results suggest that although repetitive TMS is steadily becoming the mainstay technique today, single-pulse TMS also possesses sufficient antidepressive effects. PMID- 16048449 TI - Prevalence and correlates of excessive daytime sleepiness in high school students in Korea. AB - The purpose of the present study was to determine the prevalence of excessive daytime sleepiness (EDS) and its associations with sleep habits, sleep problems, and school performance in high school students in South Korea. A total of 3871 students (2703 boys and 1168 girls with a mean age of 16.8 years and 16.9 years, respectively) aged 15-18 years in the 11th grade of high school completed a questionnaire that contained items about individual sociodemographic characteristics, sleep habits, and sleep-related problems. The overall prevalence of EDS was 15.9% (14.9% for boys and 18.2% for girls). Mean reported total sleep time was similar in EDS and non-EDS (6.4 +/- 1.6 and 6.4 +/- 1.3 h/day, respectively). The increased risk of EDS was related to perceived sleep insufficiency (P < 0.001), teeth grinding > or = 4 days/week (P < 0.001), witnessed apnea > or = 1-3 days/week (P < 0.01), nightmares > or = 4 days/week (P < 0.05), low school performance (P < 0.01), and two or more insomnia symptoms (P < 0.05). Students with low school performance had a 60% excess in the odds of EDS compared to those whose school performance was high. These findings suggest that EDS is associated with multiple sleep-related factors in adolescents. Whether interventions to modify associated correlates can alter EDS warrants consideration, especially because it may also improve academic performance in high school students. PMID- 16048450 TI - Twelve-month prevalence, severity, and treatment of common mental disorders in communities in Japan: preliminary finding from the World Mental Health Japan Survey 2002-2003. AB - To estimate the prevalence, severity, and treatment of Diagnostic and Statistical Manual of Mental Disorders (4th edn; DSM-IV) mental disorders in community populations in Japan, face-to-face household surveys were conducted in four community populations in Japan. A total of 1663 community adults responded (overall response rate, 56%). The DSM-IV disorders, severity, and treatment were assessed with the World Mental Health version of the World Health Organization (WHO) Composite International Diagnostic Interview (WMH-CIDI), a fully structured lay-administered psychiatric diagnostic interview. The prevalence of any WMH CIDI/DSM-IV disorder in the prior year was 8.8%, of which 17% of cases were severe and 47% were moderate. Among specific disorders, major depression (2.9%), specific phobia (2.7%), and alcohol abuse/dependence (2.0%) were the most prevalent. Although disorder severity was correlated with probability of treatment, only 19% of the serious or moderate cases received medical treatment in the 12 months before the interview. Older and not currently married individuals had a greater risk of having more severe DSM-IV disorders if they had experienced any within the previous 12 months. Those who had completed high school or some college were more likely to seek medical treatment than those who had completed college. The study confirmed that the prevalence of DSM-IV mental disorders was equal to that observed in Asian countries but lower than that in Western countries. The percentage of those receiving medical treatment was low even for those who suffered severe or moderate disorders. Possible strategies are discussed. PMID- 16048451 TI - Spatial working memory deficit correlates with disorganization symptoms and social functioning in schizophrenia. AB - Both spatial working memory deficit and disorganization symptoms have been considered significant components of schizophrenic impairment involved with the dorsolateral prefrontal cortex. The purpose of the present study was to investigate the relationships among spatial working memory, psychiatric symptoms including disorganization symptoms, and social functioning in schizophrenia. Fifty clinically stable patients with schizophrenia and 34 healthy controls participated in the study. Patients were rated with the Brief Psychiatric Rating Scale and the Rehabilitation Evaluation Hall and Baker. The Advanced Trail Making Test was used to evaluate spatial working memory. Patients demonstrated significantly reduced spatial working memory compared to that of healthy controls. Spatial working memory in patients correlated significantly with social functioning such as self-care skills, community skills and speech disturbance, and with disorganization symptoms. Disorganization symptoms also correlated with these aspects of social functioning. In conclusion it is suggested that both spatial working memory deficit and disorganization symptoms, which are impairments involved with the dorsolateral prefrontal cortex dysfunction, can serve as effective predictors of social functioning. PMID- 16048452 TI - Prevalence of restless legs syndrome in non-institutionalized Japanese elderly. AB - The purpose of the present paper was to evaluate the prevalence of restless legs syndrome (RLS) in a non-institutionalized Japanese elderly population. The subjects consisted of 8900 elderly people >65 years of age belonging to the Seniors Association in Izumo City in November 2000. The present study was conducted in two parts. The phase 1 investigation was a screening by mailed questionnaire and the phase 2 investigation was diagnosis by face-to-face interview. Subjects with possible cases of RLS in phase 1 proceeded to phase 2 and definite cases of RLS were then detected. In phase 1, a total of 3287 subjects completely answering all questionnaire items, were defined as the subjects of the present study. A total of 150 were classified as having 'probable RLS', resulting in a prevalence of 4.6%. These subjects with probable RLS in phase 1 were detected as the subjects of phase 2. By face-to-face interview and various clinical examinations, a total of 35 subjects (nine male, 26 female) were diagnosed as having definite RLS, resulting in a prevalence of 1.06%. Furthermore, seven subjects (two male, five female) with symptomatic RLS were detected and finally 28 subjects (seven male, 21 female) were diagnosed as having idiopathic RLS. It was significantly higher in women for both the total and idiopathic RLS groups (0.60% male vs 1.46% female; 0.46% male vs. 1.18% female, respectively). The prevalence of RLS may be lower in the Japanese elderly than that in Caucasian subjects. These results could enhance understanding of the differences in predisposition between the races. PMID- 16048453 TI - Sensory gating impairment in development of post-concussive symptoms in mild head injury. AB - Post-concussive symptoms reported by mild head injury (MHI) patients have been inadequately understood. Post-concussive symptoms reported by patients with MHI have so far been explained in terms of impairment in neurocognitive functions or deficits in modulation of flow of information. There are no studies that have looked into sensory gating impairment in MHI and its relation to post-concussive symptoms. The purpose of the present paper was to investigate the role of sensory gating impairment in post-concussive symptoms in mild head injury patients. Thirty MHI patients were evaluated for their neuropsychological functions, sensory gating deficits, and post-concussive symptoms. Neuropsychological functions were in the domain of attention, executive functions, and learning and memory. Sensory gating was assessed by Structured Interview for Assessing Perceptual Anomalies and post-concussive symptoms were assessed using the Neurobehavioral Rating Scale. Multiple regression method was used to identify predictors for post-concussive symptoms. Post-concussive symptoms were predicted by sensory gating deficits when sensory gating deficit was one of the predictors along with neuropsychological functions. Post-concussive symptoms were predicted by scores of Digit Vigilance and Digit Symbol Substitution Test, when predictors were restricted to neuropsychological functions. Sensory gating deficits were correlated with performance on Digit Symbol Substitution test. Post-concussive symptoms reported by MHI patients are the result of poor modulation of incoming sensory information. PMID- 16048454 TI - Depression and associated factors of informal caregivers versus professional caregivers of demented patients. AB - To examine the differences in depressive state and associated factors between informal and professional caregivers, a cross-sectional study was carried out in 23 informal home-based caregivers of demented patients, 24 professional caregivers working in the dementia ward of a psychiatric hospital, and 31 controls. Measurements included severity of dementia (Clinical Dementia Rating Scale, Mini Mental State Examination, MMSE) and levels of caregivers depression (Beck Depression Inventory; BDI), care burden (Zarit Caregiver Burden Interview; ZBI) and quality of life (World Health Organization-Quality of Life-26, WHO QOL26). Informal caregivers had the highest BDI score and ZBI and the lowest QOL among the three studied groups. Regarding informal caregivers, there was a strong positive correlation between BDI and ZBI scores. The BDI and ZBI scores were significantly high when patients exhibited behavioral problems. The four WHO-QOL categories (physical domain, psychological domain, social relationships and environment) had a strong negative correlation with BDI. Regression analysis demonstrated that the psychological domain points of WHO-QOL, role strain factor of ZBI and MMSE score were significantly associated with BDI. Regarding professional caregivers, their BDI score had a strong negative correlation with the physical and psychological domains and environment categories of WHO-QOL. Regression analysis demonstrated that their BDI score was significantly associated with the psychological domain and environment. It is thus very important to provide sufficient social care services and/or personal support to informal caregivers. PMID- 16048455 TI - Continuation electroconvulsive therapy for relapse prevention in middle-aged and elderly patients with intractable catatonic schizophrenia. AB - The authors have previously studied the short-term effect of the first acute electroconvulsive therapy (ECT) course (phase 1 study) on intractable catatonic schizophrenia and the 1-year relapse rate after response to the acute ECT (phase 2 study) in middle-aged and elderly patients. Results indicated that, although acute ECT has an excellent short-term effect, the 1-year relapse rate after response to acute ECT is high despite the use of continuation neuroleptics. In the present prospective study the effect was explored of continuation ECT with neuroleptics on the prevention of relapse after response to a second acute ECT course in the relapsed participants of the phase 2 study. The present study included seven consecutive patients > 45 years of age with catatonic schizophrenia (Diagnostic and Statistical Manual of Mental Disorders (4th edn; DSM-IV) who relapsed (despite the use of neuroleptics) within 1 year after response to the first acute ECT course, and then responded to the second acute ECT course. The patients were given continuation ECT combined with neuroleptics; four ECT sessions at weekly intervals, then four ECT sessions every 2 weeks, then three ECT sessions every 4 weeks. Clinical symptoms were evaluated by means of the Brief Psychiatric Rating Scale (BPRS) weekly for 48 weeks or until relapse. Relapse was defined as a BPRS score of at least 37 for 3 consecutive days. Three out of the seven patients (42.9%) had a sustained response to ECT during the 1 year follow-up period. In the seven patients the probability of relapse within 1 year under treatment with neuroleptics alone (phase 2 study) was statistically higher than that under continuation ECT combined with neuroleptics (present study). No statistical differences were seen between the phase 2 study and the present study in the severity of psychiatric symptoms, global social function, the number of acute ECT sessions or the dosage of neuroleptics. No patient experienced a severe cognitive or physical adverse effect resulting from continuation ECT. Continuation ECT with neuroleptics is an efficacious and safe treatment for maintaining a response in middle-aged and elderly patients with intractable catatonic schizophrenia who have relapsed after a positive response to acute ECT despite the use of continuation neuroleptics. PMID- 16048456 TI - Autism-Spectrum Quotient-Japanese version and its short forms for screening normally intelligent persons with pervasive developmental disorders. AB - A Japanese version of the Autism Spectrum Quotient (AQ), AQ-J was administered to 25 normally intelligent high-functioning pervasive developmental disorder (HPDD) patients (mean age, 24.2 years; 24 male, one female) and 215 controls (mean age, 30.4 years; 86 male, 129 female) randomly selected from the general population. The AQ-J had satisfactory internal consistency reliability (Cronbach's alpha > 0.70 in the two groups), test-retest reliability, and discriminant validity [i.e. the AQ-J score was significantly higher in the HPDD (mean, 29.6) than controls (mean, 22.2)]. At a cut-off of 26, the AQ-J had satisfactory sensitivity, specificity, and negative predictive value, but it had low positive predictive value (0.24) possibly due to the facts that the 25 mild HPDD patients scored lower and the controls scored higher on the AQ-J than British counterparts on the AQ. The AQ-J-21 (consisting of 21 items significantly associated with HPDD diagnosis) and the AQ-J-10 (consisting of 10 of the 21 items with an effect size > 0.17) had higher, although not satisfactory, positive predictive values of 0.35 and 0.46 at cut-offs of 12 and 7, respectively, than the AQ-J. The AQ-J and two short forms are useful not to predict but to rule out mild HPDD, the most difficult part of HPDD to be distinguished from non-PDD conditions, in persons scoring under the cut-offs and to consider professionals' examination of HPDD in persons scoring over them, because their negative predictive values were satisfactory. PMID- 16048457 TI - Mitochondrial DNA sequence analysis of patients with 'atypical psychosis'. AB - Although classical psychopathological studies have shown the presence of an independent diagnostic category, 'atypical psychosis', most psychotic patients are currently classified into two major diagnostic categories, schizophrenia and bipolar disorder, by the Diagnostic and Statistical Manual of Mental Disorders (4th edn; DSM-IV) criteria. 'Atypical psychosis' is characterized by acute confusion without systematic delusion, emotional instability, and psychomotor excitement or stupor. Such clinical features resemble those seen in organic mental syndrome, and differential diagnosis is often difficult. Because patients with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) sometimes show organic mental disorder, 'atypical psychosis' may be caused by mutations of mitochondrial DNA (mtDNA) in some patients. In the present study whole mtDNA was sequenced for seven patients with various psychotic disorders, who could be categorized as 'atypical psychosis'. None of them had known mtDNA mutations pathogenic for mitochondrial encephalopathy. Two of seven patients belonged to a subhaplogroup F1b1a with low frequency. These results did not support the hypothesis that clinical presentation of some patients with 'atypical psychosis' is a reflection of subclinical mitochondrial encephalopathy. However, the subhaplogroup F1b1a may be a good target for association study of 'atypical psychosis'. PMID- 16048458 TI - CYP2D6 gene deletion allele in patients with neuroleptic malignant syndrome: preliminary report. AB - Neuroleptic malignant syndrome (NMS) is a potentially fatal adverse reaction to psychopharmacologic treatment. Reported herein are two NMS patients with schizophrenia who were found to possess a CYP2D6 gene deletion allele (CYP2D6*5). The deletion results in decreased CYP2D6 activity, possibly leading to drug accumulation. Both patients with NMS had been treated with neuroleptics, including CYP2D6 substrates. Polymerase chain reaction (PCR) followed by restriction fragment length polymorphism analyses and long PCR were performed to detect CYP2D6 genotype. One patient was found to possess *5/*10; the other had a *1/*5 genotype. The present preliminary report suggests that pharmacokinetic factors cannot be excluded and the CYP2D6 polymorphism is possibly associated with the etiology of NMS. PMID- 16048459 TI - Assessment of pharmacological toxicity using serum anticholinergic activity in a patient with dementia. PMID- 16048460 TI - The universal importance of the 'pipeline'. PMID- 16048461 TI - Rural generalist nurses' perceptions of the effectiveness of their therapeutic interventions for patients with mental illness. AB - OBJECTIVE: To explore generalist nurses' perceptions of their efficacy in caring for mentally ill clients in rural and remote settings, and their educational needs in the area of mental health care. DESIGN: A self-administered questionnaire adapted from the Mental Health Problems Perception Questionnaire; a Likert scale used to rate the perceptions of nursing staff of their own ability to adequately treat and care for patients experiencing mental illness. SETTING: The Roma and Charleville Health Service Districts, Queensland, Australia. SUBJECTS: Nurses (Registered Nurses, Assistants in Nursing and Enrolled Nurses) in the Roma and Charleville health service districts (n = 163). MAIN OUTCOME MEASURES: Generalist nurses' perceptions regarding their therapeutic commitment, role competency and role support. RESULTS: Seventy per cent of respondents indicated that limited knowledge of mental health problems was an issue preventing nursing staff in rural and remote settings from providing optimum care to patients with mental illness. Twenty-nine per cent of respondents indicated that they had never received or undertaken training or education in relation to the care, treatment or assessment of patients with mental illness. CONCLUSION: Rural nurses do not feel competent, nor adequately supported, to deal with patients with mental health problems. In addition, the nurses' education and ongoing training do not adequately prepare them for this sphere. PMID- 16048462 TI - Perception of causes of malaria and treatment-seeking behaviour of nursing mothers in a rural community. AB - OBJECTIVE: Maternal health care workers' recognition of malaria, its transmission and treatment of children's fever at community level. DESIGN: Randomised sampling of households with children of 0-5 years. SETTING: Rural community in a forest zone. SUBJECTS: Fifty-five households at Ijegemo village, Ogun state, Nigeria were sampled and questionnaires administered to 200 mothers/care workers of children within the age group 0-5 years. Blood smears were obtained from finger pricks and level of parasitemia with malaria parasites determined for each child. MAIN OUTCOME MEASURES: Interactions with the women during the survey empowered them with accurate knowledge of malaria recognition, transmission and management of children's fever at the community level. RESULTS: Forty per cent of the respondents did not know the exact cause of malaria, 20% attributed the cause to sunlight, 16.5% to mosquitoes, 12.5% to poor hygiene, 4% to palm oil intake, 4% to blood shortage and 6% to a hot body. Of respondents 85.5% identified stagnant water as breeding site of mosquitoes but could not correlate it with the occurrence of malaria fever. Herbal concoction was the first treatment action. Some of these herbs are listed in the text. Plasmodium falciparum alone was identified in all blood smears. Children at two to three years of age were recorded with the highest percentage (67.5%) of parasite-positive cases with a mean value of 1237.04 +/- 2113.19. CONCLUSION: This study highlights a critical need for targeting health messages towards poorly educated women in order to empower them with the knowledge and resources to recognise and manage their children's health problems. PMID- 16048463 TI - Factors associated with rural doctors' intention to continue a rural career: a survey of 3072 doctors in Japan. AB - OBJECTIVE: To show the relationship between the personal and educational backgrounds of rural doctors and their intention to continue a rural career. DESIGN: Nationwide postal survey. SETTING: Public clinics or hospitals in municipalities that are classified as 'rural' by the national government. SUBJECTS: A total of 4896 doctors working for 828 public clinics and hospitals. MEASUREMENTS: A questionnaire was mailed. The questionnaire inquired about the subject's age, sex, hometown, exposure to rural practice in undergraduate education, postgraduate training, continuing medical education, current position and affiliation status with a medical school, as well as his or her intention to continue a rural career. RESULTS: Response rate was 64%; 26% answered that they intended to continue a rural career. Postgraduate training in general internal medicine, general surgery, anaesthesiology, paediatrics and gastroenterology were positively related with the intention to continue a rural career (odds ratio = 2.045, 1.59, 1.30, 1.48, 1.38). Rural background, undergraduate exposure to rural practice, multispecialty-rotation in postgraduate training and current administrative position had positive correlations with the intention to continue in logistic regression analysis (odds ratio = 1.80, 2.47, 1.54, 2.17). Affiliation with a medical school department was negatively related with the intention to continue (odds ratio = 0.45). CONCLUSION: In addition to the rural background of physicians, some undergraduate and postgraduate factors were independently associated with the intention to continue a rural career. PMID- 16048464 TI - World Assumptions as a measure of meaning in rural road crash victims. AB - OBJECTIVE: To explore the effect of experiencing a road crash on aspects of psychological meaning. DESIGN: A 12-month repeated measures prospective design was used. SETTING: Data were collected from admissions to a hospital in central Victoria. SUBJECTS: Seventy-two injured road crash victims took part in the study. MAIN OUTCOME MEASURES: Three measures of psychological trauma and the World Assumptions Scale. RESULTS: World Assumptions Scale scores did not change significantly over the year, despite improvements in other trauma scores. CONCLUSION: This study does shed some light on the psychological effects of road trauma, but further research is needed to create more sophisticated measures. PMID- 16048465 TI - Teaching clinical pathology by flexible delivery in rural sites. AB - OBJECTIVE: To evaluate the implementation of a clinical pathology and clinical skills course designed for delivery at several distributed sites, including a mandatory eight-week placement in small remote communities. DESIGN: All students in the first cohort for the course were invited to complete questionnaires rating the value of their learning experiences and the impact of relying on resources delivered by information technology. Forty-one of 63 students responded. RESULTS: Formal lectures and resources provided at the main campus were most highly valued by the students. Patient presentations in the rural and remote communities were better examples of clinical pathology than those encountered in urban hospitals, and the rural tutors were regarded very highly for their support of student learning. Delivery of resource materials in remote sites was not as successful as planned, due to difficulties with bandwidth and download speeds. Student academic performance appeared unrelated to location of learning. CONCLUSION: Students were able to achieve learning objectives for the course, relying on a richer patient mix, campus-based core sessions and information technology-based resources. Curriculum planners should be encouraged to further devolve learning in traditionally campus-based content to rural and remote communities, but only after careful planning and resource allocation to support learning in rural teaching sites. PMID- 16048466 TI - Overseas-trained doctors in Australia: community integration and their intention to stay in a rural community. AB - OBJECTIVE: The aim of this study was to identify the factors that influence foreign doctors' community integration and examine how these affect their intention to stay in the rural community. DESIGN: Qualitative study using life history perspective. SETTING: Rural communities throughout Victoria. PARTICIPANTS: Fifty-seven overseas-trained doctors (OTDs) working throughout rural Victoria, Australia. MAIN OUTCOME MEASURES: Key factors of community integration influencing OTDs' decision to stay in or leave a rural community. RESULTS: Maintaining cultural and religious values, as well as relationships to their respective ethnic communities is important to OTDs. While they do not expect excessive support from the community they appreciated the cultures of welcoming or 'embracing differences'. Supportive communication and supervisory support positively influence OTDs' appreciation of what the rural community can offer them and how they might overcome any difficulties that they face with their rural practice and life. CONCLUSION: As well as ensuring that OTDs' professional needs are met, the importance of a supportive environment within the clinic and community awareness of the OTDs' needs should not be underestimated as influences on an OTD's retention in a rural community. PMID- 16048467 TI - The diabetes experiences of Aboriginal people living in a rural Canadian community. AB - OBJECTIVE: To optimise participation with Aboriginal people by sharing experiences of living with the challenges of diabetes in rural south-western Canada, and how these could be addressed. DESIGN: Qualitative content analysis of semi-structured and conversational interviews. SETTING: Diabetes health services in the Bella Coola Valley, British Columbia, Canada. SUBJECTS: Eight Nuxalk Nation participants, five women and three men, living with type 2 diabetes, were interviewed. Four of these participants, three women and one man, were engaged in six follow-up conversational interviews. MAIN OUTCOME MEASURES: The descriptive research explored experiences of Nuxalk people living with the challenges of diabetes, and how these could inform diabetes health services in culturally specific ways. RESULTS: Challenges included understanding the connections between (i) diabetes and western or traditional medicines; (ii) dietary changes, exercise and weight loss; (iii) how health professionals communicate and the relevance of what is said; (iv) having many life choices and the responsibility to choose; and (v) a belief in living day by day and an awareness of life cycles that may need to be broken. CONCLUSION: The study substantiated the fundamental necessity for diabetes health services to be inclusive of Aboriginal perspectives. PMID- 16048468 TI - Strongyloidiasis: a review of the evidence for Australian practitioners. AB - OBJECTIVE: To summarise the available evidence concerning the prevalence, clinical manifestations, diagnosis and management of strongyloidiasis in Northern Australia. METHODS: We searched Medline, Clinical Evidence and the Cochrane Library using MeSH terms and text words 'strongyloides OR strongyloidiasis'. For Australian studies we included text words '(parasite* OR parasitic OR helminth*) AND Australia*'. We examined references contained in retrieved studies or identified from direct contact with researchers. Studies consistent with our objective that described their methods were eligible for inclusion. RESULTS: The prevalence in some tropical Aboriginal communities is high. Infection can be asymptomatic, cause a range of clinical syndromes or death. It may become chronic. Infected patients are at risk of developing severe disseminated disease particularly with immune compromise. There is little information about the relative frequency of different clinical outcomes. Available diagnostic tools are imperfect. Stool examination has a low sensitivity. Serology may have a low specificity in high prevalence populations and has not been evaluated in Aboriginal populations. Antihelmintic drugs are relatively safe and effective. Community programs based on treatment of stool-positive cases have been associated with a reduced prevalence of strongyloidiasis. We found no studies examining alternative public health interventions. CONCLUSION: There is a high prevalence in many Aboriginal communities. Strongyloides infection should be excluded prior to commencing immunosuppressive therapies in patients from endemic areas. Further studies examining the public health impact of strongyloidiasis, the role of the enzyme-linked immuno-sorbent assay serological test and population-based approaches to management of the disease in endemically infected Australian populations are needed. PMID- 16048469 TI - Qualitative evaluation of country mental health inpatient units. PMID- 16048470 TI - Awareness gained from rural experience: a student's perspective. PMID- 16048472 TI - SARRAH: give the worker tools. PMID- 16048473 TI - Critical threat to the availability of surgical implant material: lesson to be learned from breast implants. PMID- 16048475 TI - A novel insulin delivery algorithm in rats with type 1 diabetes: the fading memory proportional-derivative method. AB - An algorithm designed to automatically control insulin delivery was tested in rats with Type 1 diabetes. This nonlinear algorithm included a fading memory component of proportional and derivative errors in order to simulate normal insulin secretion. Error-weighting functions for the proportional and derivative terms were used with a performance index designed for error adaptation. In the first version of the algorithm, the proportional gain was adaptively varied. In the second version, a low rate of basal insulin delivery was adaptively varied. Six 6-h studies with each version were conducted using frequent blood sampling and intravenous insulin delivery. In Version 2 studies, blood glucose levels during the last two hours were well-controlled and significantly lower than in Version 1 (118 +/- 2.0 vs. 130 +/- 2.9 mg/dL). Neither version produced hypoglycemia. Future research using this algorithm needs to focus on automated glucose sensing in combination with insulin delivery. PMID- 16048476 TI - Expression of scavenger receptor CD36 in chronic renal failure patients. AB - BACKGROUND: Patients with chronic renal failure (CRF) are at increased risk of atherosclerosis development. One of the major steps in pathogenesis of atherosclerosis is formation of foam cells. Scavenger receptor CD36 is among the major receptors for oxidized low density lipoproteins (oxLDL) and therefore it plays a crucial role in foam cell formation. The aim of the present study was to investigate the expression of CD36 on blood monocytes of CRF patients. METHODS: Expression of CD36 on blood monocytes of CRF patients treated with hemodialysis (HD), peritoneal dialysis (PD), those not yet on dialysis (predialysis), and controls was assessed with the use of flow cytometry. Additionally, the major lipid peroxidation markers, malondialdehyde (MDA) and 4-hydroxyalkenals (HAE), were measured. Further, impact of treatment with HMG-CoA reductase inhibitors (statins) on CD36 expression in CRF patients was evaluated. RESULTS: Expression of monocyte CD36, measured as mean fluorescence intensity (MFI) was significantly higher in HD and PD patients, when compared to controls without renal insufficiency (respectively: 1011 +/- 288 and 1000 +/- 309 vs. 710 +/- 313; P < 0.01 for both groups). This was not the case in predialysis group (828 +/- 363 vs. 710 +/- 313). Higher concentrations of lipid peroxidation indicators, MDA and HAE were observed in all three subgroups of CRF patients (2.1 +/- 0.51, 2.02 +/- 0.27, and 1.81 +/- 0.53 microm in HD, PD, and predialysis group, respectively, vs. 1.13 +/- 0.59 microm in controls; P < 0.01). Patients treated with statins showed significantly lower CD36 expression than patients without statin therapy. CONCLUSIONS: The present study, for the first time, demonstrates increased expression of CD36 scavenger receptor in CRF patients. This may be a possible risk factor for accelerated atherogenesis observed in this group of patients. PMID- 16048477 TI - Predictive value of carotid intima media thickness in hemodialysis patients. AB - Atherosclerosis is accelerated in hemodialysis patients. Intima media thickness (IMT) is a strong predictor for cardiovascular events in the general population. Using B-mode ultrasonography, IMT in the common carotid arteries was measured in 99 nondiabetic hemodialysis patients (44 women and 55 men, mean age 53.1 years and mean dialysis duration 45.8 months). During a follow-up of 42.4 +/- 19.5 months, 33 patients died, 19 (57.6%) of them of cardiovascular causes. In these 19 patients IMT was significantly higher (0.89 vs. 0.69 mm) than in those who survived. Correlation between cardiovascular mortality and IMT was found. Patients were divided in relationship to the tertiles of IMT and the risk for cardiovascular death was progressively higher from the first tertile of IMT onward (P < 0.0006). IMT turned out to be an independent predictor of cardiovascular death (P < 0.025). According to our results IMT may be usefully applied for cardiovascular mortality risk stratification in nondiabetic hemodialysis patients. PMID- 16048478 TI - Computational fluid dynamics of gap flow in a biocentrifugal blood pump. AB - The centrifugal blood pump with a magnetically suspended impeller has shown its superiority as compared to other artificial heart pumps. However, there is still insufficient understanding of fluid mechanics related issues in the clearance gap. The design nature of the pump requires sufficient washout in the clearance between the impeller and the stationary pump housing inner surface. In this study, numerical simulations were carried out to investigate the flow fields in the gap of the Kyoto-NTN centrifugal blood pump. The flow patterns in the gap region of the blood pump were presented and regions of high and low velocity were identified. It was found that the radial velocity of the blood in the gap was closely related to the pressure distribution at the exit of the impeller, both the highest pressure gradient and the highest radial velocity in the gap occurred at an angular position of 170 degrees . The mass flow rate in the gap was estimated to be 25.2% of the pump outflow, which is close to the measurement results of a five times enlarged test pump. The wall shear stresses on the gap surface were found to be over 21 Pa and below 300 Pa, which is correspondingly higher than the threshold of thrombi formation and is lower than the shearing threshold of red blood cells. Comparison of the 1 : 1 simulation model with the measurement results on a five times enlarged test pump indicates that there are some differences in the resulting radial velocity distributions in the gap and thus the washout mechanism. Two symmetrical high washout regions at both the cutwater and splitter plate were observed in the simulation instead of a single washout region at the splitter plate found in the experimental study. This may be due to the scaling effect of the enlarged test pump; also the medium used in the experiment is different from the simulation. PMID- 16048479 TI - Ascending aorta outflow graft location and pulsatile ventricular assist provide optimal hemodynamic support in an adult mock circulation. AB - Although continuous flow (CF) and pulsatile flow (PF) ventricular assist devices (VADs) are being clinically used, their effects on aortic blood flow, as a measure of overall blood distribution, remain unclear. In acute VAD support animal experiments, our group has described a zone of turbulent mixing in the aortic arch. The objective of this study was to confirm this finding in the controlled setting of an adult mock circulation, simulating ventricular pathophysiologic states (normal and failing ventricle). CF and PF flow VADs were connected to ventricular apical inflow and ascending aorta (AA) or descending aorta (DA) outflow cannulae. Cardiovascular pressure and flow waveforms were recorded at varying levels of VAD bypass resulting in four test conditions: (i) CF-AA; (ii) CF-DA; (iii) PF-AA; and (iv) PF-DA. Confirming the animal data, no differences in mean aortic flow between CF and PF VADs were found, and significantly lower mean aortic arch flow with DA cannulation was noted. Mean aortic root flow decreased with increasing VAD bypass flow. As in the animal studies, despite similar mean flow rates, significant differences in waveform morphology were observed for AA and DA outflow graft locations and varying levels of VAD bypass. At 100% VAD support in the failing heart, PF restored waveform pulsatility to normal baseline while CF resulted in little pulsatility. These results confirm our earlier findings in the animal model, suggesting that outflow graft location may have a significant effect on aortic blood flow distribution. The long-term implications of these findings are being examined in ongoing studies. PMID- 16048480 TI - Effects of atrial arrhytmias on the regurgitation of a monoleaflet prosthetic heart valve. AB - Many patients who receive a prosthetic heart valve also have or acquire cardiac arrhythmias. However, most in vitro studies of prosthetic valves examine them under normal rhythms. In this study, a monoleaflet prosthetic heart valve was tested in vitro under conditions that simulated normal sinus rhythm, first degree atrioventricular heart block, and atrial fibrillation (fixed and variable ventricular rates). Atrial contraction was simulated by an active atrial chamber. The timing between the atrium and ventricle was adjusted to simulate various types of arrhythmias. The closing, leakage, and total regurgitant volumes and fractions increased for each type of atrial arrhythmia when compared to normal sinus rhythm. The peak regurgitant flow increased for first degree atrioventricular heart block and atrial fibrillation with a fixed ventricular rate compared to normal sinus rhythm. PMID- 16048481 TI - The effects of graft geometry on the patency of a systemic-to-pulmonary shunt: a computational fluid dynamics study. AB - The modified Blalock-Taussig shunt is a palliative operation for some congenital heart diseases. An artificial conduit with antithrombotic surface placed between the subclavian (or innominate) and the pulmonary artery supplies blood to the lungs in defects with decreased pulmonary flow. Clotting of the graft is the main cause of its failure. Stenosis of the arteries is also observed. The objective of the present study was to investigate the flow pattern in the graft to investigate the possibility that clotting is initiated by the stimulation of platelets by high shear stress, and the possible effect of the pathological wall shear stress on the stenosis formation. The model included the left subclavian artery (LSA), the left pulmonary artery (LPA), and the graft. The three-dimensional relative position and size of the arteries was obtained from a CT scan of real anatomy. Four different types of the graft with two different diameters (3 and 4 mm) and two different shapes (straight and curved) of the pipe, and one variable diameter pipe were inserted in the model. A pulsatile flow of 0.81 L/min on average was assumed at the inlet to LSA, and 80% of the flow was directed through the graft. Computer simulations demonstrated a complex flow pattern with eddies and low velocity regions in the arteries at the anastomoses with the graft in all five models. An eddy was also found inside the straight 4 mm graft. A high pathological shear rate was present within the graft, with higher values in the 3 mm grafts. The fractional volume with a high (>2500 L/s) shear rate was between 2.5% and 4.5%, and that with a very high (>7500 L/s) shear rate between 0 and 1.5% of the model volume, and depended on the graft geometry and the phase of the cardiac cycle. Pathologically high (>3.5 Pa) and pathologically low (<1.0 Pa) wall shear stress, which may induce neointimal growth, was found in LSA and LPA. We conclude that the activation of platelets by high shear rate is possible within the graft, followed by their subsequent aggregation in the eddies with a low flow rate. Flow-induced changes of the vessel wall thickness (stenosis) can also appear, especially in the pulmonary artery. PMID- 16048482 TI - Tissue factor, its inhibitor, and the thrombogenicity of two new synthetic membranes. AB - BACKGROUND: The aim of the study was to compare the effect of new high-flux hemodialysis membranes made from polyacrylonitrile (AN69ST) and polysulfone (Helixone) on the plasma levels of tissue factor (TF) and tissue factor pathway inhibitor (TFPI) playing a key role in hemostasis. Established thrombogenicity markers were also determined. METHODS: In a clinical prospective randomized study, 10 patients were examined using either membrane at the start and at minutes 15, 60, and 240 of hemodialysis. RESULTS: Increases in the plasma TF levels reached significance at the end of hemodialysis with both membranes, with the Helixone also after 15 min. TFPI levels tended to rise significantly from minute 15 onward while not differing from baseline at the end of the procedure. Judging by the increase in thrombin-antithrombin III complexes, both membranes significantly activated coagulation at the end of hemodialysis. Platelet factor 4 levels, released during thrombocyte and endothelial stimulation, were elevated from the start of procedures. There were no significant differences between the AN69ST and the Helixone in any of the assessed markers. CONCLUSIONS: The AN69ST and Helixone membranes do not differ in their effects on TF and TFPI or even in established thrombogenicity markers. PMID- 16048483 TI - Image processing strategies dedicated to visual cortical stimulators: a survey. AB - Multi-electrode devices are constantly evolving toward a state where complexity and reliability are adequate for providing a breakthrough in visual cortical stimulation allowing the blind to recover partial vision. Yet few research teams have focused on the development of the front-end subsystem that transforms an input image from a camera into stimulation commands. This article collects state of-the-art knowledge about the appearance and organization of phosphenes, and previous work in image processing dedicated to visual cortical stimulation. Observations and hypothesis about important issues are highlighted, and six image processing strategies that could be used in such a subsystem are presented, from the most optimistic that use brightness modulation to emulate grayscale to the most conservative that use only on/off phosphene evocation. PMID- 16048484 TI - Computational simulation of the blood separation process. AB - The aim of this work is to construct a computational fluid dynamics model capable of simulating the quasitransient process of apheresis. To this end a Lagrangian Eulerian model has been developed which tracks the blood particles within a delineated two-dimensional flow domain. Within the Eulerian method, the fluid flow conservation equations within the separator are solved. Taking the calculated values of the flow field and using a Lagrangian method, the displacement of the blood particles is calculated. Thus, the local blood density within the separator at a given time step is known. Subsequently, the flow field in the separator is recalculated. This process continues until a quasisteady behavior is reached. The simulations show good agreement with experimental results. They shows a complete separation of plasma and red blood cells, as well as nearly complete separation of red blood cells and platelets. The white blood cells build clusters in the low concentrate cell bed. PMID- 16048485 TI - The Japanese artificial organs scene: current status. AB - Artificial organs and regenerative medicine are the subjects of very active research and development (R&D) in Japan and various artificial organs are widely used in patients. Results of the R&D are presented at the annual conference of the Japanese Society for Artificial Organs (JSAO). Progress in the fields of artificial organs and regenerative medicine are reviewed annually in the Japanese Journal of Artificial Organs. The official English-language journal of JSAO, Journal of Artificial Organs, also publishes many original articles by Japanese researchers. Although the annual conference and the publications of JSAO provide the world with update information on artificial organs and regenerative medicine in Japan, the information is not always understood appropriately in the rest of the world, mainly due to language problems. This article therefore introduces the current status of artificial organs and regenerative medicine in Japan. Artificial hearts and metabolic support systems are reviewed here and other interesting areas such as regenerative medicine can be found elsewhere. PMID- 16048486 TI - The bioreactor with CYP3A4- and glutamine synthetase-introduced HepG2 cells: treatment of hepatic failure dog with diazepam overdosage. AB - A novel recombinant human hepatic cell line, CYP3A4- and glutamine synthetase (GS, an enzyme which converts ammonium ion and glutamate to glutamine)-introduced HepG2 (HepG2-GS-CYP3A4), was established. Its usefulness in a large-scale culture with a circulatory bioreactor in vitro and in dog models of ischemic hepatic failure with acute diazepam (DZP, a substrate of CYP3A4) overdosage was further examined. HepG2-GS-CYP3A4 expressed about 9 times larger amounts of CYP3A4 protein than a control. After incubation with HepG2-GS-3A4 cells in a circulatory bioreactor for 24 h, ammonia and DZP concentrations in the culture medium significantly decreased by about 40%. Furthermore, this system improved the survival time and decreased serum concentrations of DZP, ammonia, and transaminase in dogs with ischemic hepatic failure plus acute DZP overdosage. The mean survival time with bioreactor with HepG2-GS-3A4 was 42.7 +/- 3.6 h, which was significantly longer than that without reactor, with reactor (no cells), and with HepG2-GS (23.4 +/- 2.8, 22.1 +/- 2.4, and 31 +/- 3.7 h, respectively). Therefore, it is concluded that this bioartificial liver could be a good tool for the treatment of dogs with hepatic failure and that it could potentially be a bridging procedure to liver transplantation. PMID- 16048487 TI - Delta-CHr improves the identification of anemic syndromes and the evaluation of hemoglobin synthesis. AB - Reticulocyte hemoglobin content (CHr) is considered an index of iron status, helpful in the differential diagnosis of microcytoses. Its potential can be enhanced by comparing CHr dynamic reference values (CHr-e: expected CHr), which are proportional to the MCVr variations occurring in micro- or macrocytosis, with measured CHr values. We demonstrate that the difference between measured CHr and CHr-e (DeltaCHr) is helpful to differentiate the anemic syndromes and, in particular, beta-thalassemia vs. presumable sideropenia. DeltaCHr can also indicate when to interrupt iron supplementation. DeltaCHr allows an insight into the erythropoiesis of thalassemic and sideropenic subjects, pointing out the reduced hemoglobin production and ineffective erythroid activity in these conditions. PMID- 16048488 TI - Management of acute painful crises in sickle cell disease. AB - Pain is a common mode of manifestation of sickle cell disease (SCD) but there is limited information on pain management in this disorder. This study examines the use of opioids and non-opioid analgesia in the management of painful crisis in adult SCD patients; the routine use of antimalarials and antibiotics as adjunct therapy was also examined. A total of 87% of the patients had had a form of analgesics before presentation, 20% of which had parenteral analgesia. Ten per cent had not used any form of medication while another 10% used non-steroidal anti-inflammatory drugs. When asked, 59% of the patients desired oral non-opioid analgesics while 31% were not concerned about the type of analgesic given. Only 8% requested opioids. Hospital admission was not necessary in 65% of the patients; they were observed in the day-care unit and allowed home within 24 h. Sixty per cent did not have a test for malaria; 66% of those who had the test performed were negative, 35% of those whose thick film for malaria was negative had antimalarials prescribed. Only five patients (7%) were febrile at presentation. Thirty-four per cent had antibiotics prescribed, a third of these parenterally. Thirty-nine per cent had no fever but received antibiotics. PMID- 16048489 TI - Analysis and enumeration of T cells, B cells and NK cells using the monoclonal antibody fluorescence capability of a routine haematology analyser (Cell-Dyn CD4000). AB - This communication details a method for the quantitative and qualitative analysis of blood T-, B- and NK-cell populations using the Abbott Cell-Dyn CD4000 haematology analyser. A series of 66 ethylenediaminetetraacetic acid (EDTA) anticoagulated samples with lymphocyte counts between 0.2 and 33.3 x 10(9)/l were selected and analysed with CD3, CD19, Ia and CD56 monoclonal reagents. The flow cytometry reference method utilized a lymphocyte gate defined by optical scatter, with phenotypic analyses referencing to this gate and the absolute lymphocyte count. The CD4000 method analysed all leucocyte events, set primary gates for specific immunophenotypic fractions, and then determined population counts by reference to the white blood cell (WBC) count. Comparisons of CD3+ T-cell and CD19+ B-cell numbers showed high coefficients of correlation (R(2) > 0.95) and agreement (y = 1.01x) between the CD4000 and flow cytometry reference methods. Lower coefficients of correlation were obtained for CD3-CD56+ (R(2) = 0.52) and CD3+CD56+ (R(2) = 0.83) components. No major discrepancies were observed, and the CD4000 procedures additionally provided qualitative insights into the possibility of T-cell activation. The potential to undertake immediate analysis of EDTA anticoagulated blood samples to determine the nature of abnormal lymphocyte morphology or numbers represents a considerable advance in the capability of haematology laboratories. PMID- 16048490 TI - Proviral status of HTLV-1 integrated into the host genomic DNA of adult T-cell leukemia cells. AB - Human T-cell leukemia virus type-1 (HTLV-1) is the etiological agent of adult T cell leukemia (ATL), and leukemic cells always carry the proviral genome monoclonally integrated into their host genomes at the same sequence site, designated as the monoclonal integration. Using Southern blot hybridization (SBH) and sequenced tagged site polymerase chain reaction assays, we examined the proviral status in 558 clinical specimens from 350 patients who are suspected to have ATL. A total of 321 specimens (57.5%) from 241 patients showed positive results for the monoclonal integration according to SBH, using EcoR1 and Pst1. The 241 patients consisted of 136 patients (56.4%) with the complete provirus (C type), 62 patients (25.7%) with a defective provirus (D-type), and 43 patients (17.8%) with multibands (M-type). The incidence of the D- and M-types were in the order of smoldering, chronic, and acute subtypes of ATL, suggesting that such an aberrant proviral status is generated on the way to multistep carcinogenesis and is subsequently clinically important for the malignant behavior of the disease. Moreover, our data showed that the partial deletion of the proviral genome is initiated first at the site of the gag region and spreads into the sites of the pol and env regions, whereas the long terminal repeats and pX regions are almost always conserved. These results suggest that analysis of the proviral status provides useful diagnostic and virologic-oncological information about ATL and HTLV-1 pathology, especially the important role of pX gene in tumorigenesis. PMID- 16048491 TI - Persistent nucleated red blood cells in peripheral blood is a poor prognostic factor in patients undergoing stem cell transplantation. AB - We compared detection rates and counts of nucleated red blood cell (NRBC) in the peripheral blood of survivors and nonsurvivors (total 44 patients) of stem cell transplantation. The rate of NRBC detection increased to 79.5% after transplantation. After engraftment, the detection rate of NRBC decreased to 17.0% in survivors, but increased to 100% in nonsurvivors. The NRBC count increased after transplantation in both groups. This increase was transient in survivors, but increased after engraftment in nonsurvivors. The mean NRBC count after engraftment was 872 vs. 40.3 for nonsurvivors vs. survivors, respectively. At postengraftment, all patients who were negative for NRBC survived, but 10 of the 15 patients who were positive for NRBC died (66.7%). The survival rates of patients with a NRBC count >200 x 10(6)/l were significantly lower than those of patients whose counts were <100 x 10(6)/l. These data indicated that persistent NRBC in peripheral blood is a poor prognostic factor, and suggested that monitoring NRBC after SCT might provide useful clinical information. PMID- 16048492 TI - Occurrence of autoantibodies in chronic graft vs. host disease after allogeneic stem cell transplantation. AB - Chronic graft vs. host disease (GVHD) remains a major cause of morbidity and mortality after allogeneic stem cell transplantation (SCT). Chronic GVHD (cGVHD) has many similarities to de novo autoimmune disorders. While the presence and association of autoantibodies is well reported in these disorders, their role and clinical use remains a less studied area after SCT. We report the presence of autoantibodies in SCT recipients and a possible association with presence of cGVHD. During routine follow-up visits peripheral blood samples were tested for: rheumatoid factor (RF), antinuclear antibody (ANA), double stranded DNA (dsDNA), antimitochondrial antibody, antismooth muscle antibody (Anti Sm), antiendomysial, antireticulin antibodies, antithyroid peroxidase antibodies and an extractable nuclear antigen screen, in 13 SCT recipients. Six of 13 (46%) patients had one or more autoantibodies. All the patients with antibodies had cGVHD where as none of the patients without cGVHD had any autoantibodies (P = 0.025). Three (23%) patients had only one autoantibody and three (23%) of them had more than one autoantibody. ANA was positive in three (23.3%) patients, double stranded DNA in four (30.7%) patients, RF in one (7.6%) and Anti Sm muscle in two (15.3%) patients. In the present study, autoantibodies were detected predominantly in patients with presence of cGVHD. They also appeared to be more frequent in an unmanipulated graft and so less in patients with a T-cell depleted allograft. In two of 13 patients only there appeared to be an association between the antibody titre and flare up in skin symptoms. In conclusion, this small series raises interesting questions about the presence and role of autoantibodies after SCT and their association with cGVHD. PMID- 16048493 TI - Platelets in the paediatric population: the influence of age and the limitations of automation. AB - Accurate and precise platelet counting is important for the clinical management of children with platelet disorders. Current automated technologies are often unable to discriminate platelets from non-platelet particles particularly in circumstances where platelet anisocytosis is common. This study compares manual methodology and the automated technologies; impedance, optical density and CD61 immunoplatelet method (available on the Cell Dyn 4000) with the reference method of flow cytometric analysis in a paediatric population. A total of 141 samples were analysed and divided into specific age related groups and groups with thrombocytopenia and thrombocytosis. Data analysis showed that the CD61 method compared best with the reference method and this was evident in all the specified groups. The mean platelet count obtained by optical and manual methods were lower, suggesting that these methods are less reliable. The impedance count method was accurate despite its limitations. Strong correlations were observed in the 2-14 year age group but there was greater variation in the <1 month group supporting the theory that there is a greater variation in platelet characteristics in neonates. The CD61 method is the automated method of choice and would be particularly useful in the problem groups (platelet counts <50 x 10(9)/l and neonates <1 month old). PMID- 16048494 TI - Comparison of flow cytometric methods for the measurement of ZAP-70 expression in a routine diagnostic laboratory. AB - Chronic lymphocytic leukaemia (CLL) follows a variable clinical course with patient survival ranging from only a few years despite treatment, to several decades in patients who may never require clinical intervention. Determination of the mutational status of a patient's immunoglobulin heavy chain variable region (Ig V(H)) gene has been used to provide prognostic information, but this assay is not available in most laboratories. The discovery of the expression of the protein tyrosine kinase zeta-associated protein (ZAP)-70 in V(H)-unmutated CLL cases led to its proposal as a surrogate marker for V(H) status. This study investigated the measurement of ZAP-70 expression in CLL using different flow cytometric protocols. Two different antibodies and two different staining methods were compared. The Caltag ZAP-70 antibody and Fix & Perm kit were the easiest to use and were the most sensitive and specific combination, with 91% concordance between ZAP-70 expression and V(H) status. Three patients (9%) were discordant (two V(H) mutated/ZAP-70 positive, and one V(H) unmutated/ZAP-70 negative). No correlation existed between CD38 and either ZAP-70 expression or V(H) status. Measurement of ZAP-70 expression using the Caltag antibody/kit combination provides a standardized flow cytometric method that could be introduced into a routine CLL immunophenotyping panel in a clinical diagnostic laboratory. PMID- 16048495 TI - Efficacy and response to intravenous anti-D immunoglobulin in chronic idiopathic thrombocytopenic purpura. AB - This review explored the effectiveness of anti-D in the management of chronic idiopathic thrombocytopenic purpura (ITP). Of 16 patients, 14 non-splenectomized and two splenectomized, with chronic ITP received anti-D immunoglobulin at a dose of 50-75 mcg/kg. A total number of 100 doses anti-D were given. Fourteen patients had previous treatment with steroids, which was discontinued either because of unresponsiveness or unacceptably high maintenance doses. Two patients had no previous treatments with any modality. Anti-D was given as a short i.v. infusion whenever platelet count dropped below 30 x 10(9)/l or patient was haemorrhagic or preoperatively. Response was defined as an absolute platelet count >30 x 10(9)/l or an increment by > or =20 x 10(9)/l. Response was obtained in 14 patients with a response rate of 87%. Fifteen patients were not on any other form of treatment at the time of anti-D therapy and one patient had a concurrent steroid therapy. The improvement in platelet count lasted for more than 8 weeks post-57% of anti-D infusions. We report two patients with previous splenectomy for ITP who responded to anti-D therapy. The side-effects profile was very mild with no patients required red cell transfusion. PMID- 16048496 TI - Myelodysplastic syndrome associated with trisomy 2. AB - Clinical course and cytogenetic analysis suggest that myelodysplasia (MDS) is one step in a multistep model of malignant transformation of haematopoietic stem cells to acute myeloid leukaemia (AML). We report a further case of MDS associated with trisomy 2, and comment on the significance of the cytogenetic abnormality, which as a sole abnormality only occurs in MDS, but is found in combination with other chromosomal abnormalities in AML. Previous reports on balanced and unbalanced chromosomal abnormalities associated with therapy related MDS and therapy related AML suggest that trisomy 2 is an early chromosomal abnormality in leukaemogenesis. PMID- 16048498 TI - DIC secondary to acute pancreatitis. AB - Various hematological abnormalities including fall in serial values of hemoglobin or hematocrit, coagulation factor abnormalities, leukocytosis, acute hemolytic anemia, thrombocytopenia, and thrombotic thrombocytopenic purpura or hemolytic uremic syndrome have been reported in patients with acute pancreatitis. Similarly, abnormalities of blood coagulation factors consistent with disseminated intravascular coagulopathy (DIC) have also been noticed in patients with pancreatitis. We report a case of a 33-year-old female with acute pancreatitis who presented with one episode of epistaxis and abnormal prothrombin time and partial prothrombin time. Coagulation work-up revealed thrombin time 24.3 s fibrinogen 110 mg/dl, D-dimers >1 and < 2, and fibrin degradation products >22. Pancultures did not show any evidence of infection. The patient maintained a normal renal and mental status during her illness. Her D-dimers continued to decrease with resolution of acute pancreatitis as evidenced by decreased abdominal pain, relief of nausea, control of vomiting, and decrease in serum amylase and lipase levels. This case report suggests that coagulation abnormalities are encountered in patients with acute pancreatitis. It is hypothesized that such hemostatic abnormalities may be related to early intravascular consumption of coagulation factors secondary to circulating pancreatic enzymes, particularly trypsin, or secondary to vascular injury. Recognition of these hematological complications including DIC is paramount. Physicians caring for these patients should be aware of such a complication of acute pancreatitis. PMID- 16048497 TI - Two rare mutations in Turkey: IVS I.130(G-C) and IVS II.848(C-A). AB - Beta-thalassemia, an autosomal recessive disease, results from mutations of the beta-globin gene. More than 40 different mutations found in Turkish beta thalassemia patients are mostly composed of point mutations, and only in very rare cases a deletion or an insertion causes beta-thalassemia phenotypes. Here, we report two patients who were clinically diagnosed with beta-thalassemia major and HbS/beta-thalassemia respectively. We performed reverse dot blot hybridization method and automated sequence analysis to detect the mutations. One of the patients was found to be IVS I.130 (G-C) homozygous, the other was HbS/IVS II.848 (C-A) as compound heterozygous. The aim of this study was to report hematological and clinical findings in both cases related with beta-globin gene defects that are very rare. PMID- 16048499 TI - Successful treatment with rFVIIa of spontaneous intracerebral hemorrhage in a patient with mechanical prosthetic heart valves. AB - Intracerebral hemorrhage (ICH) is associated with high mortality in patients on oral anticoagulant treatment. The normalization of hemostatic balance usually requires slow-acting or risky treatments, such as vitamin K, fresh frozen plasma or prothrombin complex concentrates, which have narrow therapeutic windows particularly in cardiopathic patients like those with mechanical heart valves. Recombinant activated factor VII (rFVIIa) seems useful in patients with normal or pathologic coagulation who have an ICH. We report of a patient on acenocumarol for mitro-aortic valve replacement, referred for headache and found at computerized tomography scanning to have a parietal hemorrhage with ventricular invasion. International normalized ratio was 3.22. The patient was treated with vitamin K and with a bolus of 80 mug/kg of rFVIIa, with correction of the international normalized ratio within 15 min. Sequential computerized tomographies showed progressive reduction of the hematoma and normalization of ventricular spaces and the patient fully recovered. No valvular dysfunctions or cardiac thrombi were found at sequential echocardiograms. Neurological examination at 3 months follow-up visit was completely normal. To our knowledge, this is the first report of the safe and successful use of rFVIIa to treat ICH in a patient on oral anticoagulants for prosthetic heart valves. PMID- 16048500 TI - RET Y measurement as a sensitive indicator of iron deficiency. PMID- 16048501 TI - Proposed Sheffield quantitative criteria in cervical cytology to assist the diagnosis and grading of squamous intraepithelial lesions and dyskaryosis as the Bethesda System and British Society for Clinical Cytology definitions require amendment. PMID- 16048502 TI - Cytopathology in Europe: on seeing what the neighbours do. PMID- 16048503 TI - Proposed Sheffield quantitative criteria in cervical cytology to assist the diagnosis and grading of squamous intra-epithelial lesions, as some Bethesda system definitions require amendment. AB - OBJECTIVE: This study assesses the accuracy of published quantitative and qualitative criteria in the Bethesda System (TBS) for squamous intra-epithelial lesions. METHODS: Quantitative image analysis was undertaken on illustrations from TBS publications and also from slides in Cytology Training Centre teaching sets. Comparisons were also made with the British Society for Clinical Cytology (BSCC) terminology in cervical cytology, using the illustrations in their terminology publication and amalgamating the results into their proposed new two tier model. RESULTS: TBS quantitatively defines low-grade squamous intra epithelial lesions (LSIL) in both conventional and liquid-based cytology (LBC) preparations as showing nuclear enlargement more than x3 the area of a normal intermediate squamous cell nucleus. This study found that the increase in mean nuclear area was limited to only x2 in conventional preparations. In LBC (SurePath preparations, there was only a statistically non-significant x1.2 increase. This study identified a progressive and statistically significant reduction in mean cytoplasmic area from normal intermediate cells to LSIL and then to high-grade squamous intra-epithelial lesions (HSIL) in both conventional and LBC preparations. Furthermore, the most consistent quantitative finding in both conventional and LBC preparations was a statistically significant increase in the mean area and diameter ratios from normal intermediate cells to LSIL and then to HSIL. In all instances this varied from x2 to just below x3. This is in agreement with TBS, which states that the cytoplasmic area in HSIL is decreased leading to a marked increase in nuclear to cytoplasmic (NC) ratio. With the exception of an increase in mean nuclear area in conventional preparations from normal intermediate cells to LSIL, the predominant cause for this increase in NC ratios was a reduction in mean cytoplasmic area. The numerical increase in NC ratio for LSIL identified in this study was greater than implied by the 'slightly increased' statement in TBS. TBS comments that some HSIL cells can have the same degree of nuclear enlargement as in LSIL and that other HSIL cells may have much smaller nuclei than in LSIL. Both of these qualitative comments were supported in this study. The mean diameter NC ratios of 33% and 50% could provide useful diagnostic assistance in the distinction of normal intermediate cells and LSIL and between LSIL and HSIL, respectively. Because of overlapping individual ranges, however, additional diagnostic features such as nuclear morphology must be used in the distinction of normal intermediate cells, LSIL and HSIL. No statistical difference was identified in the mean diameter NC ratios between ASC US and LSIL in TBS publications. In addition, the proposed new BSCC low and high grades of squamous abnormality were not statistically different from ASC-US/LSIL and HSIL, respectively. This provides support that the proposed BSCC two-tier system of squamous abnormalities is comparable to TBS. This study shows that LBC has variable but major and significant effects on nuclear and cytoplasmic morphology and that quantitative definitions in conventional preparations cannot be automatically extrapolated to LBC methodology. CONCLUSIONS: The study shows that some TBS quantitative and qualitative criteria require amendment and that an alternative quantitative approach, such as diameter NC ratio has a more valid scientific evidence base. Furthermore, use of NC ratios avoids the problems associated with the variable changes in nuclear and cytoplasmic areas, occurring between conventional and different commercial LBC preparations. By contrast, classifications based on area comparisons must be tailored to the specific conventional or commercial LBC preparation. PMID- 16048504 TI - Proposed Sheffield quantitative criteria in cervical cytology to assist the grading of squamous cell dyskaryosis, as the British Society for Clinical Cytology definitions require amendment. AB - OBJECTIVE: In 1986, the British Society for Clinical Cytology (BSCC) published quantitative criteria to assist diagnosis in a three-tier grading system of squamous cell dyskaryosis. In dyskaryotic cells, area nuclear to cytoplasmic (NC) ratios below 50%, between 50% and 66% and over 66% were defined as equating with mild, moderate and severe grades respectively. Following the Terminology Conference in 2002, however, the BSCC recommended on their website that the three tier model should be replaced by a new two-tier system of low- and high-grade squamous abnormalities. The latter broadly equate with the two-grade Bethesda System (TBS) for reporting squamous intraepithelial lesions. The purpose of this study was to assess the accuracy and reproducibility of the BSCC three-tier quantitative definitions, to investigate if they were applicable to liquid-based cytology (LBC) and to see how they related to the proposed new two-tier BSCC system. METHODS: Quantitative image analysis was undertaken on illustrations from the 1986 BSCC terminology publication and on microscope slides from external quality assessment and Cytology Training Centre teaching sets. RESULTS: Analysis of mean NC ratios showed that mild, moderate and severe dyskaryosis exist as statistically different populations. Overlap of NC ratio ranges, however, limits their practical application in the three-tier model, although interestingly no overlap was noted between mild and severe dyskaryosis. No grade of dyskaryosis had a mean area NC ratio over 50%, indicating that the BSCC quantitative definitions are incorrect. The mean diameter NC ratios for mild, moderate and severe dyskaryosis were found to be 40%, 49% and 66% respectively. Accordingly it is possible that those reporting cervical cytology could be interpreting the BSCC NC ratios as meaning diameter rather than area. Amalgamation of the three-tier results into the proposed two-tier model shows that the resulting mean NC area and diameter ratios identify statistically different low- and high-grade populations. The reduced degree of overlap, however, of NC ratio ranges in the two-tier model implies that NC ratios could have a useful practical role in the separation of the low- and high-grade categories. The two categories were reasonably well separated by mean area and diameter NC ratios of 25% and 50% respectively. A two-tier model combining mild with moderate rather than severe dyskaryosis was found to be a statistically valid alternative but gave rise to NC ratios that would be difficult to use in practice. Except for moderate dyskaryosis, no significant differences were identified between the mean NC ratios of either conventional and LBC preparations or LBC preparations using two different commercial methodologies (SurePath and ThinPrep). Differences, however, were noted in area measurements between SurePath and ThinPrep and this has potential implications for classifications (such as TBS) using area comparisons as their basis. In addition, it was found that the increased NC ratio, associated with higher grades of dyskaryosis is more a consequence of progressive cytoplasmic area reduction rather than nuclear area increase. The similar NC ratios of borderline nuclear changes associated with human papilloma virus and mild dyskaryosis support the BSCC proposal that these can be combined to constitute a low-grade category. This study shows that the BSCC area NC ratio criteria of grading squamous cell dyskaryosis require amendment. In addition, this study supports the new BSCC recommendation of low- and high-grade squamous cell categories. CONCLUSIONS: The study proposes Sheffield quantitative criteria to assist the grading of squamous cell abnormalities. Quantitative diameter NC ratio measurements, however, must always be accompanied by detailed assessment of qualitative morphological features and in particular those relating to nuclear chromatin. This is equally relevant to both two- and three-tier models. PMID- 16048505 TI - Reduced expression of Claudin-7 in fine needle aspirates from breast carcinomas correlate with grading and metastatic disease. AB - OBJECTIVE: To study the immunocytochemical expression of the tight junction protein Claudin-7 in smears from breast carcinomas and correlate with grading, nodal status, locoregional and distant metastases and the cellular cohesion. METHODS: The material consisted of 52 air-dried smears from fine needle aspirates of breast carcinomas, both primary and metastatic and smears from seven benign lesions. A primary antibody to Claudin-7 was used for immunocytochemical staining. The degree of staining was recorded as negative, reduced or full, with full expression meaning equivalent to the staining pattern found in the fibroadenomas used as benign control. Staining intensity and the percentage of stained cells were evaluated. The control smears revealed a strong membrane and cytoplasmic positivity in all luminal epithelial cells. Cellular cohesion was graded as: (1) mainly cohesive groups, (2) groups and single cells and (3) mainly single cells. RESULTS: All primary and recurrent/metastatic breast lesions expressed Claudin-7. Full expression was demonstrated in 46% of the cases. Reduced expression was found in 54%. In cases with reduced expression, the percentage of stained cells were usually high, and no smear showed <50% stained tumour cells. The staining pattern was heterogeneous and always mixed membrane/cytoplasmic. Claudin-7 expression showed a significant correlation (P < 0.05) with grading, locoregional and distant metastases, nodal involvement and cellular cohesion in invasive carcinomas, but not with tumour size or subtype. CONCLUSION: Reduced expression of Claudin-7 correlated with higher tumour grade, metastatic disease, including loco-regional recurrences and with cellular discohesion. PMID- 16048506 TI - Human papillomavirus DNA detection in women with primary abnormal cytology of the cervix: prevalence and distribution of HPV genotypes. AB - OBJECTIVES: In this study, we focus on the prevalence and occurrence of different anogenital human papillomavirus (HPV) genotypes in a first abnormal cervical screening test, and correlate HPV genotyping with the cytological diagnosis on thin-layer liquid-based preparations in routine gynaecological screening. METHODS: Out of 780 abnormal smears, 513 tested positive for HPV. All 25 different HPV types were identified by Line Probe Assay. RESULTS: The prevalence of high-risk HPV types increased from 72% in atypical squamous cell of undetermined significance to 94.5% in high-grade intra-epithelial lesion (HSIL). Co-infection with multiple HPV types was predominantly found in HSIL (35.8%). In the HSIL group the most common HPV types were 16, 52, 51 and 31; type 18 was rarely present. CONCLUSION: The role of types 31, 51 and 52 should be considered in future studies on vaccine development. PMID- 16048507 TI - Neural tumours of the neck presenting as thyroid nodules: a report of three cases. AB - OBJECTIVES: Neural tumours of the neck may at times secondarily involve the thyroid and manifest clinically as thyroid nodules. On cytological evaluation these nodules may be confused with other spindle lesions of the thyroid. We report two cases of schwannoma and one case of a malignant peripheral nerve sheath tumour (MPNST) of the neck, which presented as thyroid nodules and evaluate the role of cytology in identifying these tumours. METHODS: The thyroid nodules in all the three cases were sampled by the non-aspiration technique using a 23-gauge needle. Both alcohol-fixed and air-dried smears were prepared and stained by the Papanicolaou and May-Grunwald-Giemsa stains. Cytology smears and histology sections from the resected specimens were reviewed, and the findings noted. RESULTS: Both the cases of schwannoma were correctly identified on cytology while the case of MPNST could only be typed as a spindle cell tumour. However, on cytology it was not possible to state whether the tumours were thyroidal or extrathyroidal in origin. CONCLUSIONS: Schwannomas of the neck are easily identifiable on cytology compared with MPNST. However, cytology alone is not helpful in identifying the origin of these tumours. As primary neural tumours of the thyroid are rare, the possibility of a soft tissue neural tumour extending into the thyroid should always be ruled out while evaluating these cases. PMID- 16048508 TI - Cytopathology in Finland. PMID- 16048509 TI - An unusual association with carcinoma pancreas: a case report. PMID- 16048510 TI - Children's understanding of the Ulysses conflict. AB - Two studies explored children's understanding of how the presence of conflicting mental states in a single mind can lead people to act so as to subvert their own desires. Study 1 analyzed explanations by children (4--7 years) and adults of behaviors arising from this sort of 'Ulysses conflict' and compared them with their understanding of conflicting desires in different minds, as well as with changes of mind within an individual across time. The data revealed that only the adults were able to adequately explain the Ulysses conflict. Study 2 asked children (4--7 years) and adults to choose among three explicitly presented competing explanations for self-subverting behaviors. The results suggest that an understanding of the influence of conflicting mental states on behaviors does not occur until at least 7 years of age. PMID- 16048511 TI - Reorientation by geometric and landmark information in environments of different size. AB - It has been found that disoriented children could use geometric information in combination with landmark information to reorient themselves in large but not in small experimental spaces. We tested domestic chicks in the same task and found that they were able to conjoin geometric and nongeometric (landmark) information to reorient themselves in both the large and the small space used. Moreover, chicks reoriented immediately when displaced from a large to a small experimental space and vice versa, suggesting that they used the relative metrics of the environment. However, when tested with a transformation (affine transformation) that alters the geometric relations between the target and the shape of the environment, chicks tended to make more errors based on geometric information when tested in the small than in the large space. These findings suggest that the reliance of the use of geometric information on the spatial scale of the environment is not restricted to the human species. PMID- 16048512 TI - Motor processes in children's imagery: the case of mental rotation of hands. AB - In a mental rotation task, children 5 and 6 years of age and adults had to decide as quickly as possible if a photograph of a hand showed a left or a right limb. The visually presented hands were left and right hands in palm or in back view, presented in four different angles of rotation. Participants had to give their responses with their own hands either in a regular, palms-down posture or in an inverted, palms-up posture. For both children and adults, variation of the posture of their own hand had a significant effect. Reaction times were longer the more awkward it was to bring their own hand into the position shown in the stimulus photograph. These results, together with other converging evidence, strongly suggest that young children's kinetic imagery is guided by motor processes, even more so than adults'. PMID- 16048513 TI - Auditory-visual intermodal matching of small numerosities in 6-month-old infants. AB - Recent studies have reported that preverbal infants are able to discriminate between numerosities of sets presented within a particular modality. There is still debate, however, over whether they are able to perform intermodal numerosity matching, i.e. to relate numerosities of sets presented with different sensory modalities. The present study investigated auditory-visual intermodal matching of small numerosities in infancy by using a violation-of-expectation paradigm. After being familiarized with events of a few objects impacting a surface successively, 6-month-old infants were alternatively presented with two and three tones while the movement of each object remained hidden behind an opaque screen. The screen was then removed to reveal either two or three objects. Results showed that the infants looked significantly longer at the numerically nonequivalent events (the three-tone/two-object and the two-tone/three-object events) than at the numerically equivalent events (the two-tone/two-object and the three-tone/three-object events) irrespective of the rate or duration of auditory tones presented. These findings suggest that infants are capable of performing intermodal matching of small numerosities and that they might possess abstract representations of numerosity beyond sensory modalities. PMID- 16048514 TI - Language-specific effects on number computation in toddlers. AB - A fundamental question in developmental science is how brains with and without language compute numbers. Measuring young children's verbal reactions in France (Paris) and in England (Oxford), here we show that, although there is a general arithmetic ability for small numbers that is shared by monkeys and preverbal infants, the development of such initial knowledge in humans follows specific performance patterns, depending on what language the children speak. PMID- 16048515 TI - Toddlers recognize verbs in novel situations and sentences. AB - Toddlers' (MA=22 and 27 months) ability to extend newly taught verbs to new situational and sentential contexts was investigated. Children were interactively taught two novel verbs, presented in only the transitive frame (e.g. You're lorping the ball), in a playroom setting. They then viewed the verb actions presented on side-by-side monitors and were asked to distinguish the verbs in three test frames (transitive, intransitive, neutral (e.g. lorping)). Both groups demonstrated learning of the verbs in the new situation and generalized the verbs to the intransitive sentence frame, but neither generalized reliably to the neutral frame. We conclude that even 22-month-olds demonstrate considerable context independence in their verb representations. PMID- 16048516 TI - 11-month-olds' knowledge of how familiar words sound. AB - During the first year of life, infants' perception of speech becomes tuned to the phonology of the native language, as revealed in laboratory discrimination and categorization tasks using syllable stimuli. However, the implications of these results for the development of the early vocabulary remain controversial, with some results suggesting that infants retain only vague, sketchy phonological representations of words. Five experiments using a preferential listening procedure tested Dutch 11-month-olds' responses to word, nonword and mispronounced-word stimuli. Infants listened longer to words than nonwords, but did not exhibit this response when words were mispronounced at onset or at offset. In addition, infants preferred correct pronunciations to onset mispronunciations. The results suggest that infants' encoding of familiar words includes substantial phonological detail. PMID- 16048517 TI - The genetic relationship between individual differences in social and nonsocial behaviours characteristic of autism. AB - Two types of behaviours shown in children - those reflecting social impairment and nonsocial obsessive repetitive behaviours - are central to defining and diagnosing autism spectrum disorders (ASDs). Parent and teacher data on social and nonsocial behaviours were obtained from a community sample of >3000 7-year old twin pairs. Social and nonsocial behaviours were only modestly correlated, and it was found that some individuals had extreme scores on either social or nonsocial scales but not both. Genetic model-fitting showed that social and nonsocial behaviours are both highly heritable, but their genetic overlap is modest, with most of the genetic influence being specific to either social or nonsocial behaviours. Considering these behaviours separately might help clarify gene-brain-behaviour pathways in future research. PMID- 16048518 TI - The problem of analysing the relationship between change and initial value in oral health research. AB - The relationship between initial disease status and subsequent change following treatment has attracted great interest in dental research. However, medical statisticians have repeatedly warned against correlating/regressing change with baseline because of two methodological concerns known as mathematical coupling and regression to the mean. In general, mathematical coupling occurs when one variable directly or indirectly contains the whole or part of another, and the two variables are then analyzed by using correlation or regression. Consequently, the statistical procedure of testing the null hypothesis - that the coefficient of correlation or the slope of regression is zero - may become inappropriate. Regression to the mean occurs with any variable that fluctuates within an individual or a population, either owing to measurement error and/or to physiological variation. The aim of this article was to clarify the conceptual confusion around mathematical coupling and regression to the mean within the statistical literature, and to correct a popular misconception about the correct analysis of the relationship between change and initial value. As examples that use inappropriate methods to analyze the relationship between change and baseline are still found in leading dental journals, this article seeks to help oral health researchers understand these problems and explain how to overcome them. PMID- 16048519 TI - The relationship between baseline value and its change: problems in categorization and the proposal of a new method. AB - Oral health researchers have shown great interest in the relationship between the initial status of diseases and subsequent changes following treatment. Two main approaches have been adopted to provide evidence of a positive association between baseline values and their changes following treatment. One approach is to use correlation or regression to test the relationship between baseline measurements and subsequent change (correlation/regression approach). The second approach is to categorize the lesions into subgroups, according to threshold values, and subsequently compare the treatment effects across the two (or more) subgroups (categorization approach). However, the correlation/regression approach suffers a methodological weakness known as mathematical coupling. Consequently, the statistical procedure of testing the null hypothesis becomes inappropriate. Categorization seems to avoid the problem of mathematical coupling, although it still suffers regression to the mean. We show, first, how the appropriate null hypothesis may be established to analyze the relationship between baseline values and change in the correlation approach and, second, we use computer simulations to investigate the impact of regression to the mean on the significance testing of the differences in the average treatment effects (or average baseline values) in the categorization approach. Data available from previous literature are reanalyzed by testing the appropriate null hypotheses and the results are compared to those from testing the usual (incorrect) null hypothesis. The results indicate that both the correlation and categorization approaches can give rise to misleading conclusions and that more appropriate methods, such as Oldham's method and our new approach of deriving the correct null hypothesis, should be adopted. PMID- 16048520 TI - Oral Impacts on Daily Performance in Norwegian adults: validity, reliability and prevalence estimates. AB - The Oral Impacts on Daily Performance (OIDP) instrument was translated into Norwegian and reviewed for cultural and conceptual equivalence by a group of bilingual academics. A sample of employees from the University of Bergen completed the Norwegian OIDP frequency questionnaire twice. A total of 173 and 108 subjects participated in the first and the second administration, respectively, of this questionnaire. A two-stage proportionate random sample, comprising 2,000 residents (age-range 16-79 yr), was drawn from the national population register by the Central Bureau of Statistics. Information became available for 1,309 persons who completed telephone interviews. The Norwegian OIDP preserved the overall concept of the English version. Test-retest reliability, in terms of Cohen's kappa, was 0.65, and Cronbach's alpha was high (> or = 0.80). In both samples, variations in the OIDP scores were apparent in relation to self-reported oral health and number of remaining teeth, supporting construct and criterion validity of the inventory. Only three of the OIDP interviews were discarded, which supports face validity. A total of 18.3% confirmed that they had at least one oral impact. Age-specific rates were 17.5%, 19.0%, 17.9% and 18.4% among 16-24, 24-44, 45-66 and 67-79-yr-old participants. The satisfactory psychometric properties provide evidence for the cross-cultural use of the OIDP. The presence of a distinct floor effect indicates poor sensitivity of the OIDP to detect improvements of oral health-related quality of life at a population level. Prevalence estimates were low, suggesting that the current oral health status has little impact on the daily performance of the Norwegian adult population. PMID- 16048521 TI - Dental fear, regularity of dental attendance and subjective evaluation of dental erosion in women with eating disorders. AB - This questionnaire study, with a response rate of 53%, examined self-induced vomiting, erosions and dental attendance in women with eating disorders (EDs) as well as dental fear and its effect on attendance and communication with the dentist. A survey of 371 responding women with EDs, who were recruited from a self-help organization, revealed that dental fear was higher in women with EDs compared to the general population. Dental fear was present in 32.1% of women with EDs, and very high dental fear was present in 16.5% of women with EDs. Of those with very high dental fear, 32.3% had not visited a dental clinic at all in the preceding 2 yr, and 43.5% only initiated contact when they had symptoms. Self induced vomiting was especially frequent in women with bulimia nervosa (87.9%) and in those with more than one ED (the 'mixed group') (80.6%). Among those with self-induced vomiting, 45.3% thought that they had erosions, although only 28.4% had erosions diagnosed by a dentist. Of women with EDs, 61.4% failed to disclose their condition. High dental fear did not affect willingness to disclose the ED. We conclude that dentists should examine ED patients carefully for dental erosions. Moreover, they should realize that most ED patients avoid disclosing their disorder and that dental fear further complicates dental treatment in these patients. PMID- 16048522 TI - Morphological appearance and chemical composition of enamel in primary teeth from patients with 22q11 deletion syndrome. AB - Patients with 22q11 deletion syndrome have many and complex medical problems, including hypocalcemia and/or hypoparathyroidism. Odontological findings include enamel aberrations in both dentitions. In order to describe enamel morphology, chemical composition in primary teeth, and to investigate the relationship between medical history and morphological appearance, dental enamel was investigated in 38 exfoliated primary teeth from 15 children and adolescents. Morphology was studied by the use of a polarized light microscope, microradiography, scanning electron microscopy, X-ray microanalysis, and secondary ion mass spectrometry. The morphological findings were compared with medical history. The teeth showed, in principle, a normal morphological appearance with regard to prism structure. A high frequency of aberrations, such as hypomineralization, hypoplasia and extra incremental lines, were found. The majority of the aberrations were found around the neonatal line. There was a relationship between high numbers of medical problems in the patients and enamel deviations. The result supports the hypothesis of under-reporting of both hypocalcemia and hypoparathyroidism in patients with 22q11 deletion syndrome. PMID- 16048523 TI - Activation of the Notch signaling pathway in response to pulp capping of rat molars. AB - Notch signaling is an evolutionarily conserved pathway that controls the developmental choices made by individual cells. Cells communicate via Notch receptors and their ligands, which direct decisions on the fate of stem cells according to the states of their neighbors. In this study we explored Notch signaling after the pulp capping of adult first upper rat molars. The wound was capped with calcium hydroxide. In situ hybridization revealed an increased expression of Notch signaling genes on day 1, which showed a tendency to decrease on day 3. Notch1 increased in the subodontoblast zone and close to the lesion limited to a few cells. Notch2 increased in pulp stroma surrounded by coronal odontoblasts. Notch1 and, especially, Notch3 expression increased, corresponding to perivascular cell groups. A low increase of ligand expression was observed near the injury with Delta1 expression along the dentin wall and Jagged1 in the stroma. Expression of the downstream target, Hes1, was observed along the lesion and adjacent dentin walls. Hes5 expression was not observed. The results indicate that Notch signaling is activated in response to injury and associated with the differentiation of pulp cells into perivascular cells and odontoblasts. The findings are consistent with the concept that the Notch pathway controls stem cell fate during pulp regeneration. PMID- 16048524 TI - Bone morphogenetic protein 3 expression pattern in rat condylar cartilage, femoral cartilage and mandibular fracture callus. AB - Mandibular condylar cartilage differs from primary cartilage in morphological organization of the chondrocytes and in responses to biomechanical stress and humoral factors. For the first time, we describe the expression of Bmp3 mRNA in relation to types I, II and X collagen mRNA (as determined by in situ hybridization) in chondrocytes of the rat mandibular condylar cartilage, femoral articular cartilage, femoral growth plate cartilage, and temporal cartilage, which transiently appeared in the reparative response stage of mandibular ramus fracture healing. In all cartilages evaluated, Bmp3 was expressed in proliferating chondrocytes that expressed type I collagen in condylar cartilage, articular cartilage, and temporal cartilage appearing during fracture healing. Bmp3 was also found in hypertrophic chondrocytes that expressed type X collagen mRNA in all cartilages evaluated. Furthermore, in remodeling bone, Bmp3 mRNA was strongly expressed in active osteoblast cells in periosteal reaction layers formed after fracture. These findings suggest that Bmp3 expression in a special layer of typical articular cartilage may be regulated by mechanical stress stimulation. We also found that Bmp3 was expressed in the periosteal layers of the bone segments near the fracture site during fracture healing. PMID- 16048525 TI - Microbubble-induced detachment of coadhering oral bacteria from salivary pellicles. AB - The presence and maturity of the salivary pellicle influences microbial adhesion and its tenacity in the oral cavity, posing a challenge to different plaque control systems. Some plaque-control systems rely on surface-tension forces arising from passing microbubbles sprayed over the pellicle. Passage of such bubbles is accompanied by a high fluid flow, but systematic studies are lacking on the contribution of fluid flow vs. microbubbles towards plaque removal. Therefore, the aim of this study was to determine the detachment efficacy of laminar fluid flow (wall shear rates 11,000-16,000 s(-1)), with and without microbubbles, towards the detachment of Actinomyces naeslundii T14V-J1 and Streptococcus oralis J22, and their coadhering aggregates, from salivary pellicles formed over 2 h or 16 h from reconstituted human whole saliva. Microbubbles in a fluid flow were more efficient at inducing single bacterial detachment, resulting in almost complete (97%) removal for S. oralis J22 and a 15 fold increase in A. naeslundii T14V-J1 removal as compared to the detachment caused by fluid flow alone. A. naeslundii was more difficult to remove and apparently formed the strongest bonds with high-molecular-weight proteins in 16-h pellicles. The detachment of coaggregates after 2 min left a substantial amount of adhered bacterial mass, including more than 60% of singly attached A. naeslundii on the pellicle surface, providing nucleation sites for the de novo adhesion of coadhering streptococci. PMID- 16048526 TI - Biogeneric tooth: a new mathematical representation for tooth morphology in lower first molars. AB - A mathematical representation of tooth morphology may help to improve and automate restorative computer-aided design processes, virtual dental education, and parametric morphology. However, to date, no quantitative formulation has been identified for the description of dental features. The aim of this study was to establish and to validate a mathematical process for describing the morphology of first lower molars. Stone replicas of 170 caries-free first lower molars from young patients were measured three-dimensionally with a resolution of about 100,000 points. First, the average tooth was computed, which captures the common features of the molar's surface quantitatively. For this, the crucial step was to establish a dense point-to-point correspondence between all teeth. The algorithm did not involve any prior knowledge about teeth. In a second step, principal component analysis was carried out. Repeated for 3 different reference teeth, the procedure yielded average teeth that were nearly independent of the reference (less than +/- 40 microm). Additionally, the results indicate that only a few principal components determine a high percentage of the three-dimensional shape variability of first lower molars (e.g. the first five principal components describe 52% of the total variance, the first 10 principal components 72% and the first 20 principal components 83%). With the novel approach presented in this paper, surfaces of teeth can be described efficiently in terms of only a few parameters. This mathematical representation is called the 'biogeneric tooth'. PMID- 16048527 TI - In vivo degradation of resin-dentin bonds produced by a self-etch and an etch-and rinse adhesive. AB - Resin-dentin bonds are known to degrade in the relatively aggressive oral environment. In order to obtain greater insight into the interfacial degradation process, we examined, by using transmission electron microscopy (TEM), the interfacial ultrastructure of two adhesives bonded to dentin after 1 yr in vivo. Class V cavities were prepared on the buccal surfaces of 14 intact teeth of two monkeys and then restored by using either the two-step self-etch adhesive, Unifil Bond, or the two-step etch-and-rinse adhesive, Single Bond, in combination with the restorative microhybrid composite, Z250. After 1 yr, 10 other teeth were restored by using the same materials (controls). One day later, the monkeys were killed, following which the microtensile bond strength to dentin was determined and the interfacial ultrastructure was examined by TEM. Whereas no noticeable changes in the morphology of the resin-dentin interface were observed between the 1-d and 1-yr specimens for Unifil Bond, Single Bond exhibited signs of interfacial degradation, in particular at the bottom of the 3 microm-deep hybrid layer. In conclusion, the adhesive interface produced by the etch-and-rinse adhesive was less resistant to degradation than that produced by the self-etch system. PMID- 16048528 TI - The diversity of prevention and health promotion services offered by Quebec Community Health Centres: a study of infant and toddler programmes. AB - Research has shown that preventive and health promotion (PHP) services offered by community health centres are often vulnerable in times of health reform or budgetary cutbacks. The present study was conducted in Quebec, Canada, approximately 10 years after the start of a major transformation of health care services. The objectives were: (1) to examine the diversity of infant/toddler prevention and health promotion services offered by centres locaux de services communautaires (CLSCs, local community health and social services centres); and (2) to investigate the organisational and environmental correlates of such diversity. All Quebec CLSCs were invited to complete a survey questionnaire regarding their PHP services (response rate = 69%). Data on environmental and organisational factors were extracted from existing Quebec government data sets. Bivariate and multivariate analyses were performed to identify the correlates of the CLSCs' level of diversity of PHP services. The results revealed that CLSCs varied greatly in terms of their diversity of PHP services. The number of full time equivalents in the Child and Family Services Programme (one of two programmes aimed at reaching the targeted clientele) and the degree of CLSC collaboration with other community-based organisations in the planning and delivery of PHP activities were positively associated with the level of diversity, whereas the unemployment rate in the population served was a negative predictor. This study makes an important contribution in being one of the very few which contribute to building our knowledge about the performance of community health centres. Identifying the correlates of diversity among PHP services offered by CLSCs helps us to begin to understand the processes at work in the production of PHP activities, as well as the potential leveraging forces. PMID- 16048529 TI - Young adults' (16-25 years) suggestions for providing developmentally appropriate diabetes services: a qualitative study. AB - Managing the multiple demands of a chronic condition whilst negotiating the developmental tasks of adolescence and young adulthood is a process that is neither well described nor understood, particularly in relation to providing developmentally appropriate health care for young people. The importance of this issue is starting to be reflected within the literature, and although research into models of service delivery is emerging, a lack of user involvement in service development is apparent. This qualitative, user involvement study aimed to describe and understand the considered opinions of 19 young adults with diabetes who were receiving secondary care services about the provision of diabetes services for young people. The findings, gathered using semistructured interview and focus group methods, have potentially wide-reaching implications across primary and secondary health care, and across agencies providing services to children and young people, in terms of facilitating a person's transition through adolescence and into young adult life. Participants suggested key issues to address when developing services for young people, including staff consistency, civility, clinic structures which help a person navigate the health care system, provision of age-specific information, and support in relation to a range of health, emotional, social and developmental needs. Health care professionals can help young people to meet the expectations upon them as autonomous service users by modelling appropriate relationships, helping them to acquire skills and knowledge, and overcome barriers to them becoming active participants in their health care and achieving social participation in a fuller sense. It is somewhat arbitrary to delineate between adolescence and young adulthood in terms of age alone, but in this paper, 'adolescence' refers to the period between 11 and 15 years of age, and 'young adulthood' between 16 and 25 years of age. The phrase 'young people' will also be used to refer to people between 11 and 25 years. PMID- 16048530 TI - The last 3 months of life: care, transitions and the place of death of older people. AB - Many older people die in hospitals, whereas research indicates that they would prefer to die at home. Little is known about the factors associated with place of death. The aim of the present study was to investigate the care received by older people in the last 3 months of their life, the transitions in care and the predictors of place of death. In this population-based study, interviews were held with 270 proxy respondents to obtain data on 342 deceased participants (79% response rate) in the Longitudinal Aging Study Amsterdam. In the last 3 months of life, the utilisation of formal care increased. Half of the community-dwelling older people and their families were confronted with transitions to institutional care, in most cases to hospitals. Women relied less often on informal care only, and were more dependent than men on institutional care. For people who only received informal care, the odds of dying in a hospital were 3.68 times the odds for those who received a combination of formal and informal home care. The chance of dying in a hospital was also related to the geographical region. The authors argue that future research is needed into the association that they found in the present study, i.e. that decedents who received both formal and informal care were more likely to die at home. In view of the differences found in geographical region in relation to place of death, further investigation of regional differences in the availability and accessibility of care is indicated. PMID- 16048531 TI - From patients to providers: prospects for self-care skills trainers in the National Health Service. AB - Changes in the National Health Service (NHS) workforce and user involvement have been key areas of contemporary health care policy. Potentially, the current separation of the agendas for patient participation and reconfiguration of the workforce are being brought closer together in the NHS through the Expert Patients Programme (EPP), and its potential to create a new community health 'workforce' of self-management and self-care skills trainers and tutors. The aim of the present paper is to assess the establishment and prospects of these trainers as a new workforce role in the EPP and the NHS. This is done through policy analysis and process evaluation recording the development and implementation of the EPP in the NHS. Telephone interviews with individual trainers were undertaken in order to identify the way in which they are being introduced into the NHS. A representative sample of 19 trainers from the 26 teams covering all the primary care trusts (PCTs) in England were interviewed. The EPP trainers are employed and appointed by the Department of Health and assist PCTs to run EPP courses. The analysis of trainer perspectives illuminated some of the tensions inherent in this new role, which emerge from the consequences of having a long-term condition, their relationship to other occupations operating within primary care and their structural position within the NHS. Prospects for the future development of self-management trainers remain uncertain. Two likely outcomes are examined. The first is that trainers and volunteer tutors will become increasingly integrated as a semi-professionalised group within the primary care workforce. The other option is for trainers to become freelance consultants commissioned by PCTs to run local self-management, self-care support or EPP programmes with the consequence of establishing a more distant relationship with mainstream NHS provision. PMID- 16048532 TI - Families' experiences of caring for technology-dependent children: a temporal perspective. AB - In the present study, families' experiences of caring for a technology-dependent child were examined from a temporal perspective. This involved exploring the multiple 'technological', 'social' and 'natural' rhythms and routines around which the families' lives were variously structured. A purposive sample of 36 families with technology-dependent children who used one or more medical devices on a daily basis was recruited. Devices included feeding pumps, suction machines, dialysis machines and ventilators. Using mainly qualitative methods, children, parents and siblings were interviewed to establish what the care routines involved and how these impacted on family members. The authors found that the rhythms and routines of care varied across the sample, depending on the type and number of devices used, the individual child's needs, and who provided technical care during the day and/or at night at home and in other settings. While the children's health and quality of life benefited from the technology, the time demands of the care routines and lack of compatibility with other social and institutional timeframes had some negative implications for the children and their families, limiting their participation in school, employment and social life in general. The need to use and oversee the use of some medical technologies at night also meant that many parents suffered regular disruption to their sleep. In conclusion, the authors argue that the care of technology-dependent children at home places considerable time demands on families. Families have little or no access to suitably trained carers who can provide technical care required in the home or away from the home to give parents and the whole family a break from caring where required. More trained carers and short-term care provision, better coordination of services and improvements in the design of devices would all help to reduce the negative effects of the care routines on families. PMID- 16048533 TI - Exploring the 'fit' between people and providers: refugee health needs and health care services in Mt Roskill, Auckland, New Zealand. AB - The needs of refugees and the struggles on the part of service providers to address this diverse population have received limited attention within the academic literature. This paper profiles Hauora o Puketapapa/Roskill Union and Community Health Centre (HoP), which is a non-profit, community owned and operated health clinic designed to deliver accessible, affordable and appropriate primary health care services to low-income groups in the Mt Roskill area of Auckland, New Zealand. The clinic's locality has undergone considerable demographic change over recent years with the arrival of refugees from diverse backgrounds. This situation has resulted in new sets of health needs and expectations which need to be addressed. The study took place in 2002-2003 and employed qualitative methods. In-depth interviews with community representatives, clinic users and health service staff members revealed that refugees face considerable barriers in accessing and utilising health services. Similarly, we found that health practitioners face the daunting task of endeavouring to meet these needs in an effective and culturally appropriate manner within a limited funding environment. We conclude that, despite these challenges, HoP has successfully established itself as a well-regarded place of primary health care. In so doing, it has strengthened the capacity of the local community to respond to the changing policy environment. However, long-term sustainability issues remain unless resourcing issues are adequately addressed. PMID- 16048534 TI - The challenges associated with providing community care for people with complex needs in rural areas: a qualitative investigation. AB - The aim of the present study was to explore the experiences of recipients and providers of community care in rural areas in Northern Ireland. Additionally, the authors sought to examine the impact of location, housing and environmental factors on the delivery of community care to older people with complex needs. Individual, semistructured interviews were held with service users (n = 17) and family carers (n = 14). Individual and focus group interviews were conducted with care assistants, health and social care professionals, and senior managers from a large health and social care trust and health and social services board in Northern Ireland. The importance of enabling older people to remain in their own homes and communities was emphasised by all participants. The main challenges associated with care provision in rural areas included: difficulties recruiting care assistants; lack of choice of care assistants; isolation; travel and distance between clients and their care assistants; and poor housing conditions. There was a general consensus among participants that the effectiveness of rural community care was perceived to be reliant upon the goodwill of the community. Additionally, changing demographic trends and the predicted shortfall in the number of formal and informal carers were considered key issues for service planners. A number of creative strategies could be used to address many of the limitations associated with rural isolation. These should involve capitalising on available community networks. However, planners should also acknowledge that additional resources are required to maintain older people in rural communities. PMID- 16048535 TI - The health consequences of multiple roles at older ages in the UK. AB - Increasing proportions of men and women are combining family (including care giving) and work responsibilities in later life; however, the relationship between multiple role commitments and health at older ages remains unclear. Employing data from the longitudinal Retirement Survey (1988-1989 and 1994), the present authors applied logistic regression models to investigate the relationship between multiple role occupancy (1) cross-sectionally in 1988-1989 and health status in 1994; (2) retrospectively over the respondent's lifetime up to 1988-1989 and health outcomes in 1988-1989; and (3) retrospectively between 1988-1989 and 1994, and health status in 1994. The health outcomes considered were (1) general health status, (2) functional ability and (3) severity of disability category. Overall, simultaneous role occupancy (e.g. care-giving and employment) at older ages does not appear to be associated with poor health. The authors report a positive association between employment and health, as expected. There were mixed results concerning the association between care-giving and health. Where adverse health outcomes were found, the parental role, alone or in combination with other roles, was most frequently related to poor health. Thus, for a nationally representative sample of mid-life men and women, the combination of care-giving with other family and work roles appears to have few negative health consequences. Further research is needed on whether continued parental demands in mid-life have a negative impact on health. PMID- 16048536 TI - Desired outcomes for children and young people with complex health care needs, and children who do not use speech for communication. AB - Within recent policies and initiatives, there is a growing emphasis on outcomes focused practice and user-defined outcomes to ensure that the children and families most at risk of social exclusion have every opportunity to build successful and independent lives. However, we still know little about what children and young people with disabilities consider constitutes a good quality of life. The present paper reports the findings of a qualitative research project that aimed to identify the desires and aspirations of children and young people with complex health care needs (CHNs), as well as those who do not use speech for communication (NS) and their parents. Fifty families participated in this study; 26 families had a child with CHNs and 24 had a child with NS. Semistructured interviews were carried out with 50 parents, 18 children and 17 other informants (teachers and other non-parent carers who knew the child well). A range of visual or non-verbal techniques were used to facilitate interviews with children. The research found that, while children and their parents aspired to the same sort of outcomes as those of non-disabled children, such as to have friends, interests and future independence, the specific meaning they attached to certain outcomes and the way they prioritised aspirations differed. These findings have implications for the ways in which outcomes are defined and measured, and the role of services in achieving these outcomes. PMID- 16048538 TI - Mucosal immunity. PMID- 16048539 TI - Organizing a mucosal defense. AB - Gastrointestinal associated lymphoid tissue can be divided into loosely organized effector sites, which include the lamina propria and intraepithelial lymphocytes, and more organized structures, such as mesenteric lymph nodes (LNs), Peyer's patches (PPs), isolated lymphoid follicles, and cryptopatches (CPs). These organized structures in the gastrointestinal tract have been hypothesized to play the role of primary lymphoid organ, supporting the extrathymic development of T lymphocytes (CPs), secondary lymphoid organs involved in the induction of the mucosal immune response (PPs), and tertiary lymphoid structures whose function is still under debate (isolated lymphoid follicles). The most widely studied lymphoid structure found in the small intestine is the PP. PPs are secondary lymphoid structures, and their development and function have been extensively investigated. However, single lymphoid aggregates resembling PPs have been also described in humans and in the murine small intestines. These isolated lymphoid follicles have both germinal centers and an overlying follicle-associated epithelium, suggesting that they also can function as inductive sites for the mucosal immune response. This review compares and contrasts the development and function of the four main organized gastrointestinal lymphoid tissues: CPs, isolated lymphoid follicles, PPs, and mesenteric LNs. PMID- 16048540 TI - Bronchus- and nasal-associated lymphoid tissues. AB - The bronchus-associated lymphoid tissue (BALT) and the nasal-associated lymphoid tissue (NALT) constitute organized lymphoid aggregates that are capable of T- and B-cell responses to inhaled antigens. BALT, located mostly at bifurcations of the bronchus in animals and humans, is present in the fetus and develops rapidly following birth, especially in the presence of antigens. Humoral immune responses elicited by BALT are primarily immunoglobulin A secretion both locally and by BALT-derived B cells that have trafficked to distant mucosal sites. Similarly located T-cell responses have been noted. On the basis of these findings, the BALT can be thought of as functionally analogous to mucosal lymphoid aggregates in the intestine and is deemed a member of the common mucosal immunologic system. NALT has been described principally in the rodent nasal passage as two separate lymphoid aggregates. It develops after birth, likely in response to antigen, and B- and T-cell responses parallel those that occur in BALT. It is not known whether NALT cells traffic to distant mucosal sites, although mucosal responses have been detected after nasal immunization. NALT appears from many studies to be a functionally distinct lymphoid aggregate when compared with BALT and Peyer's patches. It may exist, however, in humans as a diffuse collection of isolated lymphoid follicles. PMID- 16048541 TI - Mucosal B cells: phenotypic characteristics, transcriptional regulation, and homing properties. AB - Mucosal antibody defense depends on a complex cooperation between local B cells and secretory epithelia. Mucosa-associated lymphoid tissue gives rise to B cells with striking J-chain expression that are seeded to secretory effector sites. Such preferential homing constitutes the biological basis for local production of polymeric immunoglobulin A (pIgA) and pentameric IgM with high affinity to the epithelial pIg receptor that readily can export these antibodies to the mucosal surface. This ultimate functional goal of mucosal B-cell differentiation appears to explain why the J chain is also expressed by IgG- and IgD-producing plasma cells (PCs) occurring at secretory tissue sites; these immunocytes may be considered as 'spin-offs' from early effector clones that through class switch are on their way to pIgA production. Abundant evidence supports the notion that intestinal PCs are largely derived from B cells initially activated in gut associated lymphoid tissue (GALT). Nevertheless, insufficient knowledge exists concerning the relative importance of M cells, major histocompatibility complex class II-expressing epithelial cells, and professional antigen-presenting cells for the uptake, processing, and presentation of luminal antigens in GALT to accomplish the extensive and sustained priming and expansion of mucosal B cells. Likewise, it is unclear how the germinal center reaction in GALT so strikingly can promote class switch to IgA and expression of J chain. Although B-cell migration from GALT to the intestinal lamina propria is guided by rather well defined adhesion molecules and chemokines/chemokine receptors, the cues directing preferential homing to different segments of the gut require better definition. This is even more so for the molecules involved in homing of mucosal B cells to secretory effector sites beyond the gut, and in this respect, the role of Waldever's ring (including the palatine tonsils and adenoids) as a regional inductive tissue needs further characterization. Data suggest a remarkable compartmentalization of the mucosal immune system that must be taken into account in the development of effective local vaccines to protect specifically the airways, eyes, oral cavity, small and large intestines, and urogenital tract. PMID- 16048542 TI - Mucosal immunoglobulins. AB - Due to their vast surface area, the mucosal surfaces of the body represent a major site of potential attack by invading pathogens. The secretions that bathe mucosal surfaces contain significant levels of immunoglobulins (Igs), which play key roles in immune defense of these surfaces. IgA is the predominant antibody class in many external secretions and has many functional attributes, both direct and indirect, that serve to prevent infective agents such as bacteria and viruses from breaching the mucosal barrier. This review details current understanding of the structural and functional characteristics of IgA, including interaction with specific receptors (such as Fc(alpha)RI, Fc(alpha)/microR, and CD71) and presents examples of the means by which certain pathogens circumvent the protective properties of this important Ig. PMID- 16048543 TI - The polymeric immunoglobulin receptor: bridging innate and adaptive immune responses at mucosal surfaces. AB - Secretory antibodies of the immunoglobulin A (IgA) class form the first line of antigen-specific immune protection against inhaled, ingested, and sexually transmitted pathogens and antigens at mucosal surfaces. Epithelial transcytosis of polymeric IgA (pIgA) is mediated by the polymeric immunoglobulin receptor (pIgR). At the apical surface, the extracellular ligand-binding region of pIgR, known as secretory component (SC), is cleaved and released in free form or as a component of secretory IgA (SIgA). SC has innate anti-microbial properties, and it protects SIgA from proteolytic degradation. Expression of pIgR is regulated by microbial products through Toll-like receptor signaling and by host factors such as cytokines and hormones. Recent studies of the structure of the extracellular ligand-binding domain of pIgR have revealed mechanisms by which it binds pIgA and other ligands. During transcytosis, pIgA has been shown to neutralize pathogens and antigens within intracellular vesicular compartments. The recent identification of disease-associated polymorphisms in human pIgR near the cleavage site may help to unravel the mystery of how pIgR is cleaved to SC. The identification of novel functions for SC and SIgA has expanded our view of the immunobiology of pIgR, a key component of the mucosal immune system that bridges innate and adaptive immune defense. PMID- 16048544 TI - Lymphocyte homing to the gut: attraction, adhesion, and commitment. AB - Lymphocytes continuously migrate from the blood into the intestine. Naive lymphocytes leave the blood through high endothelial venules in Peyer's patches. During the multistep extravasation cascade, they sequentially roll on, firmly adhere to, and transmigrate through the endothelial layer using multiple adhesion molecules and chemotactic signals. In the organized lymphoid tissues of the gut, lymphocytes can become activated, if they meet their cognate antigens transported to Peyer's patches through the gut epithelium. During activation and proliferation, the lymphocytes become imprinted by the local dendritic cells, so that after returning to systemic circulation via the efferent lymphatic vasculature, they preferentially home to lamina propria of the gut to execute their effector functions. In inflammation, the recirculation routes of lymphocytes are altered, and these may explain the pathogenesis of certain extra intestinal manifestations of gut infections and inflammatory bowel diseases. The increased knowledge on the mechanisms that regulate lymphocyte homing and imprinting has clear applicability in designing more effective vaccination regimens. A detailed understanding of the mucosal homing has recently led to the development of the first successful anti-adhesive therapeutics in human. PMID- 16048545 TI - IELs: enforcing law and order in the court of the intestinal epithelium. AB - The intestinal intraepithelial lymphocytes (IELs) are mostly T cells dispersed as single cells within the epithelial cell layer that surrounds the intestinal lumen. IELs are, therefore, strategically located at the interface between the antigen-rich outside world and the sterile core of the body. The intestine of higher vertebrates has further evolved to harbor numerous commensal bacteria that carry out important functions for the host, and while defensive immunity can effectively protect against the invasion of pathogens, similar immune reactions against food-derived antigens or harmless colonizing bacteria can result in unnecessary and sometimes damaging immune responses. Probably as a result of this unique dilemma imposed by the gut environment, multiple subsets of IEL have differentiated, which all display characteristics of 'activated yet resting' immune cells. Despite this common feature, IELs are heterogeneous with regard to their phenotype, ontogeny, and function. In this review, we discuss the different subtypes of IELs and highlight the distinct pathways they took that led to their unique differentiation into highly specialized effector memory T cells, which provide the most effective immune protection yet in a strictly regulated fashion to preserve the integrity and vital functions of the intestinal mucosal epithelium. PMID- 16048546 TI - Involvement of intestinal dendritic cells in oral tolerance, immunity to pathogens, and inflammatory bowel disease. AB - Dendritic cells (DCs) are composed of a family of cells, now recognized to be essential for innate and acquired immunity. DCs at mucosal surfaces have a particular capacity to induce the differentiation of regulatory T cells producing interleukin-10 (IL-10) and transforming growth factor-beta (TGF-beta) in the steady state (non-infected, non-immunized), yet they retain the capacity to induce effector T cells in response to invasive pathogens. This decision between the induction of active immunity and tolerance will depend on the subpopulation of DC involved and the surface receptors engaged during DC activation and T-cell priming. The local microenvironment will likely play an important role both in defining the DC phenotype and in providing direct signals to responding T cells. Furthermore, DCs in organized mucosal lymphoid tissues preferentially induce the expression of CCR9 and alpha4beta7 on T cells, which results in T-cell homing to the intestinal lamina propria. Finally, DCs may play an important role in the maintenance of abnormal intestinal inflammation either by driving pathogenic T cell responses in mesenteric lymph nodes or by acting to expand or maintain pathogenic T cells locally at sites of inflammation. In this review, a brief discussion of general issues of DC biology that are pertinent to mucosal immunity is followed by a more in-depth discussion of the phenotype and function of DC populations in the intestine. PMID- 16048547 TI - Intestinal macrophages: unique effector cells of the innate immune system. AB - The gastrointestinal mucosa is the largest reservoir of macrophages in the body. These important effector cells are derived from blood monocytes that are recruited to the lamina propria by endogenous chemoattractants in the non inflamed mucosa and by inflammatory chemokines and bacterial products during inflammation. In the non-inflamed mucosa, newly recruited pro-inflammatory monocytes are exposed to lamina propria stromal (extracellular matrix) factors that induce phenotypic and functional differentiation into non-inflammatory macrophages. As a consequence of this differentiation, resident lamina propria macrophages are strikingly downregulated for the expression of innate response receptors, such as the receptors for lipopolysaccharide, immunoglobulin G (IgG), and IgA, and the production of pro-inflammatory cytokines, including interleukin 1 (IL-1), IL-6, IL-8, and tumor necrosis factor-alpha. Despite downregulated pro inflammatory function, strong phagocytic and bactericidal activities remain intact. Thus, in the non-inflamed intestinal mucosa, lamina propria macrophages are non-inflammatory but retain avid scavenger and host defense functions, a unique but ideal phenotype and functional profile for effector cells in close proximity to immunostimulatory microorganisms and products. PMID- 16048548 TI - Non-classical MHC class I molecules on intestinal epithelial cells: mediators of mucosal crosstalk. AB - The mucosal immune environment consists of a complex combination of lymphoid cells, non-lymphoid cells, and lumenal bacteria. Signals from lumenal bacteria are constantly transmitted to the underlying tissues across the intestinal epithelial barrier. Intestinal epithelial cells (IECs) can sense these signals, integrate them, and interpret them for lamina propria lymphoid populations. One mechanism by which these signals are communicated is by the expression of non classical major histocompatibility complex (MHC) class I molecules by IECs. Epithelial cells can express a surprising variety of non-classical MHC class I molecules. In some cases, IECs can act as non-professional antigen-presenting cells utilizing the expression of such non-classical MHC class I molecules to directly present bacterial antigens. In other cases, the expression of non classical MHC class I molecules may act as a co-stimulatory molecule or adhesion molecule that can modify the mucosal immune response. Finally, the expression of these molecules on IECs can lead to a broad array of responses ranging from tolerance to inflammation. Overall, the IEC, via the expression of non-classical MHC class I molecules, is a central mediator of the constant crosstalk between the intestinal lumen and the mucosal immune system. PMID- 16048549 TI - Development of intestinal M cells. AB - Intestinal epithelium contains several specialized cell types including M cells, which can be found in the follicle-associated epithelium (FAE) or occasionally on the villi. M cells are critical for sampling of intestinal flora and for transferring pathogens across the epithelial barrier for recognition by the immune system. Development of M cells on the villi (M(v)) is independent of the presence of lymphocytes, while development of the FAE and M cells within the FAE (M(f)) is dependent on B lymphocytes. Here, the concept is discussed that B cells are not required for induction of M(f) differentiation but are required for transition to and maintenance of the mature M(f) phenotype. Signaling pathways possibly involved in the B-cell-independent stages of M-cell development are also discussed. PMID- 16048550 TI - Rapidly changing perspectives about mast cells at mucosal surfaces. AB - Mast cells (MCs) are major effector cells of immunoglobulin E (IgE)-mediated allergic inflammation. However, it has become increasingly clear that they also play important roles in diverse physiological and pathological processes. Recent advances have focused on the importance of MCs in both innate and adaptive immune responses and have fostered studies of MCs beyond the myopic focus on allergic reactions. MCs possess a variety of surface receptors and may be activated by inflammatory mediators, IgE, IgG, light chains, complement fragments, proteases, hormones, neuropeptides, and microbial products. Following activation, they produce a plethora of pro-inflammatory mediators and participate in inflammatory reactions in many organs. This review focuses on the role of MCs in inflammatory reactions in mucosal surfaces with particular emphasis on their role in respiratory and gastrointestinal inflammatory conditions. PMID- 16048551 TI - Mucosal immunity and allergic responses: lack of regulation and/or lack of microbial stimulation? AB - Allergic hyperreactivity is defined as an exaggerated immune response [typically immunoglobulin E (IgE) but also non-IgE mediated] toward harmless antigenic stimuli. The prevalence of allergic disease has increased dramatically during the last 20 years, especially in developed countries. Both genetic and environmental factors contribute to susceptibility to allergy. Evidence has emerged supporting the hypothesis that a reduction in antigenic stimulation brought about by widespread vaccination, improvements in standards of hygiene, and extensive use of antibiotics has contributed to the dysregulation of T-helper 2 cell (Th2) type responsiveness that typifies allergy. Regulation of the inherently Th2-biased mucosal immune response is crucial both to the maintenance of homeostasis at this strategic defensive barrier and to the prevention of allergic disease. The ability of Th1 responses to counter-regulate Th2 reactivity is well characterized. More recently, interest has centered on regulatory T cells, which can suppress both Th1 and Th2 cells through the secretion of immunosuppressive cytokines such as interleukin-10 and transforming growth factor-beta. In this review, we discuss the basic cellular mechanisms of allergic diseases at mucosal surfaces, focusing on allergic responses to food, before examining newer work that suggests the induction of allergic hyperreactivity is due to a deficient immunoregulatory network, a lack of microbial stimulation, or both. PMID- 16048552 TI - Innate and adaptive immunity: the yin and yang of celiac disease. AB - Celiac disease is a multigenetic complex inflammatory disorder with an autoimmune component, induced by gluten, a protein found in wheat. It is a unique human disease model to dissect the innate and adaptive immune mechanisms underlying T cell-mediated tissue destruction and the development of T-cell lymphoma in conditions of chronic T-cell activation. PMID- 16048554 TI - Experimental models of inflammatory bowel disease reveal innate, adaptive, and regulatory mechanisms of host dialogue with the microbiota. AB - There are now many experimental models of inflammatory bowel disease (IBD), most of which are due to induced mutations in mice that result in an impaired homeostasis with the intestinal microbiota. These models can be clustered into several broad categories that, in turn, define the crucial cellular and molecular mechanisms of host microbial interactions in the intestine. The first of these components is innate immunity defined broadly to include both myeloid and epithelial cell mechanisms. A second component is the effector response of the adaptive immune system, which, in most instances, comprises the CD4+ T cell and its relevant cytokines. The third component is regulation, which can involve multiple cell types, but again particularly involves CD4+ T cells. Severe impairment of a single component can result in disease, but many models demonstrate milder defects in more than one component. The same is true for both spontaneous models of IBD, C3H/HeJBir and SAMPI/Yit mice. The thesis is advanced that 'multiple hits' or defects in these interacting components is required for IBD to occur in both mouse and human. PMID- 16048553 TI - Oral tolerance. AB - Multiple mechanisms of tolerance are induced by oral antigen. Low doses favor active suppression, whereas higher doses favor clonal anergy/deletion. Oral antigen induces T-helper 2 [interleukin (IL)-4/IL-10] and Th3 [transforming growth factor (TGF)-beta] T cells plus CD4+CD25+ regulatory cells and latency associated peptide+ T cells. Induction of oral tolerance is enhanced by IL-4, IL 10, anti-IL-12, TGF-beta, cholera toxin B subunit, Flt-3 ligand, and anti-CD40 ligand. Oral (and nasal) antigen administration suppresses animal models of autoimmune diseases including experimental autoimmune encephalitis, uveitis, thyroiditis, myasthenia, arthritis, and diabetes in the non-obese diabetic (NOD) mouse, plus non-autoimmune diseases such as asthma, atherosclerosis, graft rejection, allergy, colitis, stroke, and models of Alzheimer's disease. Oral tolerance has been tested in human autoimmune diseases including multiple sclerosis (MS), arthritis, uveitis, and diabetes and in allergy, contact sensitivity to dinitrochlorobenzene (DNCB), and nickel allergy. Although positive results have been observed in phase II trials, no effect was observed in phase III trials of CII in rheumatoid arthritis or oral myelin and glatiramer acetate (GA) in MS. Large placebo effects were observed, and new trials of oral GA are underway. Oral insulin has recently been shown to delay onset of diabetes in at risk populations, and confirmatory trials of oral insulin are being planned. Mucosal tolerance is an attractive approach for treatment of autoimmune and inflammatory diseases because of lack of toxicity, ease of administration over time, and antigen-specific mechanisms of action. The successful application of oral tolerance for the treatment of human diseases will depend on dose, developing immune markers to assess immunologic effects, route (nasal versus oral), formulation, mucosal adjuvants, combination therapy, and early therapy. PMID- 16048555 TI - Defects in mucosal immunity leading to Crohn's disease. AB - Crohn's disease (CD) is characterized by patchy transmural inflammation involving any part of the intestinal tract. Animal models have provided a great deal of insight into the pathogenesis of CD, but no animal model has recapitulated the full spectrum of manifestations witnessed in human disease. The defects in mucosal immunity in CD can be divided into those that involve the epithelial barrier, those that involve the innate immune response, and finally, defects in the adaptive immune response. Defects in the epithelial barrier in CD include an increase in intestinal permeability, increased adherence of bacteria, and decreased expression of defensins. Murine and human studies have demonstrated an increased expression of T-helper 1 (Th1) cytokines by lamina propria lymphocytes. This increased Th1 cytokine expression is driven by interleukin-12 (IL-12)/IL-23 and tumor necrosis factor-like 1A (TL1A) production by antigen-presenting cells, resulting in Tbet expression by CD4+ T cells. Another dimension of the inappropriate immune response in CD is T-cell and B-cell reactivity to luminal microbes. With the identification of the nucleotide-binding oligomerization domain 2 (NOD2) gene as a susceptibility gene, defects in the innate immune response are beginning to be explored. One may consider a model in which defective innate immune clearance of pathogens or commensal bacteria in CD leads to an inappropriate adaptive immune response to the commensal flora. PMID- 16048556 TI - Defects in mucosal immunity leading to ulcerative colitis. AB - Evidence accumulated over the last decade demonstrates that what we call 'ulcerative colitis' is actually a heterogeneous group of diseases resulting from different pathogenic mechanisms with a common symptomatic expression. Subgroups of patients with ulcerative colitis can be stratified by presence or absence of serum autoantibodies, which are thought not to be pathogenic but to mark for a distinct disease phenotype. In recent years, animal-based experimental systems have emerged that reflect human ulcerative colitis and have potential to accelerate our understanding of its pathogenesis. Genetic and immunological data from human studies in combination with results from animal model systems are the foundation of a hypothesis, which includes a role for microbial antigen exposure in the initiation, perpetuation, and amplification of the disease. In ulcerative colitis, it appears as though the T-cell response to the antigens is not T-helper (Th) 1 dominant as in the case of Crohn's disease but rather is either Th2 [interleukin (IL)-4, IL-13] or is mediated by specialized cells such as natural killer (NK) T cells (IL-13). Lamina propria T cells from ulcerative colitis patients produce significantly greater amounts of IL-13. Ulcerative colitis is associated with an atypical Th2 response mediated by a distinct subset of NK T cells that produce IL-13 and are cytotoxic for epithelial cells. The way in which this response affects the ultimate cascade of inflammatory events has yet to be determined. PMID- 16048557 TI - Innate and adaptive immunity in female genital tract: cellular responses and interactions. AB - The mucosal immune system in the female reproductive tract (FRT) has evolved to meet the unique requirements of dealing with sexually transmitted bacterial and viral pathogens, allogeneic spermatozoa, and the immunologically distinct fetus. Analysis of the FRT indicates that the key cells of the innate and adaptive immune systems are present and functionally responsive to antigens. Acting through Toll-like receptors in the Fallopian tubes, uterus, cervix, and in the vagina, epithelial cells, macrophages, natural killer cells, and neutrophils confer protection through the production of chemokines and cytokines, which recruit and activate immune cells, as well as bactericidal and virucidal agents, which confer protection at times when adaptive immunity is downregulated by sex hormones to meet the constraints of procreation. The overall goal of this paper is to define the innate immune system in the FRT and, where possible, to define the regulatory influences that occur during the menstrual cycle that contribute to protection from and susceptibility to potential pathogens. By understanding the nature of this protection and the ways in which innate and adaptive immunity interact, these studies provide the opportunity to contribute to the foundation of information essential for ensuring reproductive health. PMID- 16048559 TI - Integration of hepatitis B virus DNA and hepatocellular carcinoma. PMID- 16048560 TI - Significance of RCAS1 antigen in hepatocellular, cholangiocellular and pancreatic carcinomas. AB - Since receptor-binding cancer antigen expressed on SiSo cells (RCAS1) was first reported as a tumor-associated antigen of gynecologic cancer, its function and practical value as a tumor marker have been investigated in various types of carcinomas. Basic research has indicated that RCAS1 expression in cancer cells contributes to the evasion from immune surveillance and progression of carcinomas. The clinical significance of RCAS1 expression in hepatobiliary and pancreatic carcinomas has also been investigated. In this review, we summarize the clinical application of RCAS1 antigen in hepatic and pancreaticobiliary diseases. We present new data and review current knowledge about the potential of RCAS1 as a tumor marker and the relationship between RCAS1 expression and clinicopathologic parameters. We found that the clinical function of RCAS1 appeared to differ according to the type of carcinoma. In hepatocellular carcinoma, the clinical significance of histological RCAS1 expression was controversial and that of serum RCAS1 levels showed little clinical value. In pancreaticobiliary cancers, high RCAS1 expression in tissue samples was an unfavorable independent prognostic factor. Serum RCAS1 was a superior tumor marker reflecting the disease activity in biliary carcinoma. In pancreatic cancer, serum RCAS1 levels were less useful than in biliary carcinoma but may be available for genetically CA19-9-negative patients and for CA19-9-non-producing cancer. This review also offers suggestions for future studies. PMID- 16048561 TI - Azathioprine and 6-mercaptopurine pharmacogenetics and metabolite monitoring in inflammatory bowel disease. AB - The thiopurine drugs azathioprine and 6-mercaptopurine (6-MP) are well established in the treatment of inflammatory bowel disease (IBD). However, there is a wide inter- and intra-patient variation in the concentrations of active and toxic metabolites due to their complex metabolism and genetic polymorphisms in metabolizing enzymes. Serious drug toxicity leads to cessation of therapy in 9 25% of patients, and there is failure to achieve efficacy in approximately 15% of cases. Advances in the understanding of thiopurine drug metabolism have led to new genetic and metabolite tests to help clinicians optimize thiopurine use. Thiopurine methyltransferase (TPMT) enzyme activity can predict life-threatening myelotoxicity in the one in 300 patients who are TPMT-deficient. However, myelotoxicity can also occur in the presence of normal TPMT activity so blood count monitoring should remain standard practice. TPMT testing may also aid in dose individualization. 6-Thioguanine nucleotides (6-TGN) are thought to be the predominant active metabolites of the thiopurines. 6-thioguanine nucleotide concentration is correlated with bone marrow toxicity and may also correlate with efficacy in IBD. Measurement of 6-TGN and 6-methylmercaptopurine (6-MMP) concentration is most useful in determining why a patient is not responding to a standard dose of a thiopurine drug and may help in avoiding myelosuppression. The ratio of these metabolites can help distinguish non-compliance, under-dosing, thiopurine-resistant and thiopurine-refractory disease. Some of these investigations are entering routine clinical practice but more research is required to determine their optimal use in patients with IBD. PMID- 16048562 TI - Vasoconstrictor responses are normal but prostanoid-mediated vasodilatation is enhanced in human cirrhotic mesenteric arteries. AB - BACKGROUND AND AIMS: The mechanisms responsible for mesenteric vasodilatation in cirrhosis have not been fully elucidated. The aim of the present study was to examine whether there is altered intrinsic vascular reactivity of human mesenteric vessels in cirrhosis, which might contribute to vasodilatation in vivo. METHODS: Ten mesenteric arteries from six cirrhosis patients undergoing liver transplantation were compared with 11 arteries from six control patients. Vasoconstrictor responses to potassium, norepinephrine and methoxamine were determined. Endothelium-dependent vasodilatation responses to acetylcholine and substance P were determined both before and after inhibition of nitric oxide (NO) and prostanoid synthesis. RESULTS: Cirrhotic vessels responded normally to potassium depolarization and did not differ to control vessels with respect to sensitivity and maximal response to norepinephrine. In cirrhotic vessels, inhibition of NO synthesis had significantly less effect on substance P-induced vasorelaxation than in controls (% Relaxation: cirrhosis 70.3 +/- 9.6; control 34.9 +/- 9.5; P = 0.03). However, after inhibition of both NO and prostanoid synthesis, vasodilatory responses were eliminated in both groups. CONCLUSIONS: The findings of the present study indicate that intrinsic hyporesponsiveness to vasoconstrictors does not play a pathogenetic role in the mesenteric vasodilatation in human cirrhosis. Furthermore, vasodilator prostanoids might make a significant contribution in mediating enhanced endothelium-dependent vasorelaxation in the mesenteric circulation. PMID- 16048563 TI - Influence of load of hepatitis A virus on disease severity and its relationship with clinical manifestations in patients with hepatitis A. AB - BACKGROUND AND AIM: The purpose of the present study was to investigate the influence of viral load on disease severity and analyze the possible relationship of the load of hepatitis A virus (HAV) with disease severity and laboratory findings. METHODS: Fifty-eight patients diagnosed with acute hepatitis A were used in the current study, of whom 12 patients progressed to severe acute hepatitis (s-AH) defined on the basis of a prothrombin time (PT) of <40% and 46 patients were diagnosed as having mild acute hepatitis (m-AH). The load of HAV was measured with real-time polymerase chain reaction. RESULTS: Peak viral load showed a significant correlation with alanine aminotransferase (ALT) (r = 0.363, P = 0.0048) and PT levels (r = -0.330, P = 0.0110). In terms of disease severity, there was a significant correlation with ALT (r = 0.462, P = 0.0012) and PT levels (r = 0.400, P = 0.0059) in the m-AH group, but not in the s-AH group. A significant positive correlation of peak viral load with the C-reactive protein level (r = 0.270, P = 0.0400) and a significant negative correlation of peak viral load with the platelet count (r = -0.313, P = 0.0015) was also found. CONCLUSIONS: The load of HAV was closely correlated with liver damage and disease severity in m-AH, but not in s-AH. The load of HAV was also closely associated with the increase in C-reactive protein level and enhancement of thrombocytopenia. PMID- 16048564 TI - Gallstone disease in non-alcoholic fatty liver: prevalence and associated factors. AB - BACKGROUND: Insulin resistance is a risk factors for non-alcoholic fatty liver disease (NAFLD) and for gallstone disease (GD). Aims of the present study were to assess the prevalence of and factors associated with GD in unselected patients with NAFLD. METHODS: A total of 161 consecutive patients with NAFLD diagnosed through compatible ultrasonography in the absence of known etiologies of liver disease (in all patients) and/or confirmed histologically (in 61 patients), was studied. Gallstone disease was diagnosed through ultrasound scanning or on the basis of previous cholecystectomy. Anthropometric and biochemical variables and concurrent diseases were compared in 32 NAFLD-GD patients and in 129 NAFLD patients without GD (controls) according to gender. RESULTS: The overall prevalence of GD was 19.88%, higher in female patients (P < 0.05), who were older (P < 001). The overall percentage of GD increased with age (P < 0.05). The GD patients had higher uric acid (men), total cholesterol and apolipoprotein B (apo B) serum concentrations (women; P < 0.05); women also had a higher prevalence of hypertriglyceridemia (P < 0.05). The age-corrected odds ratio of having GD by tertiles increased significantly with increasing uric acid (men) and with increasing total cholesterol, triglycerides and apo-B (women). At univariate continuous analysis GD was associated with insulin 120 min and uric acid in male patients; and with body mass index, insulin 120 min, apo-B, total cholesterol and triglycerides in female patients. On multivariate analysis it was found that among these factors only uric acid in men and apo-B in women were independently associated with GD in NAFLD. CONCLUSIONS: The prevalence of GD in NAFLD is more elevated than reported in the general population. The factors independently associated with GD in NAFLD are different from those reported in the general population and vary according to the gender. PMID- 16048565 TI - 249ser p53 mutation in the serum of black southern African patients with hepatocellular carcinoma. AB - BACKGROUND AND AIMS: A specific mutation at codon 249 of the p53 tumor suppressor gene (guanine to thymine; arginine to serine [249(serine)p53]) is present in the cell-free plasma of 30-47% of patients with hepatocellular carcinoma (HCC) in regions with uniformly high levels of dietary exposure to the fungal toxin, aflatoxin B(1). No information is available from other regions. We therefore examined cell-free serum from HCC patients in southern Africa, where aflatoxin B(1) exposure ranges from very high to low levels. METHODS: DNA extracted from the serum of 158 black African patients with HCC was amplified by the polymerase chain reaction assay using primers specific for exon 7 of the p53 gene, and submitted to endonuclease cleavage with HaeIII to identify the 249(serine)p53 mutation. The presence of the mutation was confirmed by nucleotide sequencing. RESULTS: The specific mutation was detected in 18% of the patients, giving an odds ratio for HCC in those with the mutation of 13.3 (95% confidence limits 1.8; 100.2). Surprisingly, the mutation was present equally often in rural and urban patients, despite presumed levels of aflatoxin B(1) exposure in the latter being much lower. No correlation was found with the presence of hepatitis B virus infection or the age, sex or tribe of the patients. CONCLUSIONS: The 249(serine)p53 mutation is found less often in the serum of patients with HCC in a region with variable levels of exposure to aflatoxin B(1) than in those with uniformly high levels of exposure, but the mutation does occur in black Africans with presumed lower levels of exposure to the fungal toxin. PMID- 16048566 TI - Genetic polymorphisms of cytochrome P450 in patients with hepatitis C virus associated hepatocellular carcinoma. AB - BACKGROUND: The carcinogenic process can be modulated by exposure to endogenous or environmental substance(s) acting as carcinogens or protocarcinogens. Polymorphic enzymes of cytochrome P450 (CYP) that play a role in detoxication/toxication of such substances via metabolization may account for the interpatient variability of clinical course in cancers such as hepatocellular carcinoma (HCC). Many CYP genetic polymorphisms, which may change enzyme activity, are known to exist in Japanese. The aim of the present study was to compare the frequencies of CYP polymorphisms between hepatitis C virus (HCV) related HCC patients and healthy subjects. METHODS: Seven mutant alleles and related genotypes of CYP in 44 HCV-positive HCC patients were chosen as follows: *1C heterozygous, *1C homozygous and *1F homozygous for CYP1A2, *4A homozygous for CYP2A6, *2A or *3 heterozygous, *2A or *3 homozygous and *2A and *3 heterozygous for CYP2C19, and *10/*5 homozygous for CYP2D6. These mutant alleles have been reported to change the CYP enzyme activity in Japanese. The frequencies of the mutant alleles and genotypes were then compared with those reported in healthy Japanese. RESULTS AND CONCLUSION: There is no statistically significant difference in genetic mutant alleles between the two groups, except for the genotype of CYP2A6*4A homozygous. The frequency of this genotype in the HCC patients (0.144) is significantly higher than that in healthy Japanese (0.034; P < 0.05; odds ratio 3.36). The clinical significance related to HCC is unknown. Further evaluation of CYP2A6*4A (deletion type) in HCV-related HCC patients is required. PMID- 16048567 TI - Relationship between expression of cyclin D1 and impaired liver regeneration observed in fibrotic or cirrhotic rats. AB - BACKGROUND AND AIM: The mechanisms responsible for impaired regenerative ability after hepatic resection observed in chronic liver disease are not fully understood. We have examined the relationships between an altered expression of cell cycle-related proteins in regenerating liver after partial hepatectomy and the impaired regenerative process observed in fibrotic and cirrhotic rats. METHODS: We performed 70% partial hepatectomy in both control and porcine serum induced fibrotic rats, and 45% partial hepatectomy in thioacetamide-induced cirrhotic rats because of the high mortality associated with 70% partial hepatectomy. Liver regeneration was monitored by proliferating cell nuclear antigen labeling index and the expression of G1 regulatory cell cycle-related proteins was determined by immunoblot analysis. RESULTS: Compared with controls, hepatocyte DNA synthesis, and induction of cyclin D1 and p21(CIP1) proteins were delayed but not suppressed in porcine serum-induced fibrotic rats and markedly inhibited in thioacetamide-induced cirrhotic rats. p27(KIP1) protein levels were unaffected by partial hepatectomy and did not differ among all three groups. CONCLUSION: Two distinct rat models of liver fibrosis and cirrhosis showed markedly different proliferative responses after partial hepatectomy. The delay or failure of cyclin D1 induction, but not the increase of p21(CIP1) or p27(KIP1) might be responsible for their impaired liver regeneration. PMID- 16048568 TI - Differential integration rates of hepatitis B virus DNA in the liver of children with chronic hepatitis B virus infection and hepatocellular carcinoma. AB - BACKGROUND AND AIM: Integration of hepatitis B virus-DNA (HBV-DNA) into the host genome, a phenomenon found frequently in hepatocellular carcinomas (HCC) and causally linked to oncogenesis, has not been well characterized in children. The aim of the present study was to determine the prevalence of HBV integration more accurately and to decide whether the integration rate varies at different stages of chronic HBV infection in children. METHODS: Of 13 children with chronic hepatitis, 14 liver biopsy tissues were analyzed. One liver tissue with pure liver cirrhosis, nine non-tumor, and nine tumor liver tissues from children with HCC were analyzed by a very sensitive method, inverse polymerase chain reaction (IPCR). RESULTS: Thirteen genuine viral-host junctional sequences from 23 patients were successfully isolated and proved that IPCR is a useful method in this context. The results also indicated that the detection rate of HBV-DNA integration increased in parallel with the progress of liver histology towards the neoplastic transformation, with 0% in the liver of chronic hepatitis, 22.2% in non-tumor livers of HCC patients, and 66.7% in tumor liver tissues of HCC patients. CONCLUSION: The present results indicate that integration of HBV-DNA into the host genome was rarely confirmed at the early stage of chronic hepatitis in children until the stage of HCC formation. PMID- 16048569 TI - Long-term clinical outcome of large volume paracentesis with intravenous albumin in patients with spontaneous bacterial peritonitis: a randomized prospective study. AB - BACKGROUND AND AIM: Large volume paracentesis (LVP) with plasma volume expansion has been used for tense or refractory ascites. However, still in question is whether it is safe and effective for the treatment of spontaneous bacterial peritonitis (SBP). We addressed this issue and conducted a study to assess safety and long-term outcome of LVP in cirrhotic patients with SBP. METHODS: Forty-two randomly assigned cirrhotic patients with SBP were classified into two groups; Group 1 included 21 patients who were treated with LVP and intravenous albumin; and Group 2 included 21 patients who were treated with diuretics and intravenous albumin. RESULTS: The overall cumulative survival rate was poor in patients with SBP (42.5% and 22.5% at 6 and 12 months, respectively). At 7 days after treatment, the blood tests were similar between the two groups. In the ascitic fluid, the white blood cell counts decreased significantly and the protein concentrations tended to increase in both groups. In-hospital days, resolution rate of SBP, and in-hospital mortality rate were similar between the two groups. Although complication rates tended to be slightly higher in Group 1, long-term cumulative survivals were similar between Group 1 and Group 2. LVP was effective in removing abdominal discomfort in patients with tense ascites without serious complication. CONCLUSIONS: LVP with intravenous albumin was as effective as diuretics with intravenous albumin for the treatment of SBP with similar mortality. LVP with intravenous albumin might be feasible for the treatment of tense or refractory ascites in cirrhotic patients with SBP. PMID- 16048570 TI - Factors affecting the mortality of pediatric fulminant hepatic failure in relation to hepatitis B virus infection. AB - AIM: To investigate the factors affecting the outcome of fulminant hepatic failure (FHF) in children in relation to hepatitis B virus (HBV) infection. METHODS: Retrospective review of a total of 94 cases (61 males and 33 females, aged from 1 month to 15 years) recruited from nine tertiary referral centers in Taiwan from 1985 to 1999. RESULTS: The overall mortality rate was 75%. Patients in the mortality group were of an older age, had higher peak total bilirubin levels, a longer prothrombin time, and a lower percentage of HBV positivity (P < 0.001, P = 0.003, P = 0.0027 and P = 0.042, respectively). Mortality was 65% in the HBV positive (n = 42) and 83% in the HBV negative (n = 52) group (P = 0.05). In the HBV positive group, the prothrombin time was noted to be the single factor affecting outcome (P = 0.036). In the HBV negative group, older age and higher peak value of total serum bilirubin were suggestive of poor survival rate (P < 0.001 and P = 0.006, respectively). Multivariate analysis revealed that total bilirubin was the single factor affecting outcome in the HBV-negative group. The mortality rate of HBV positive children in three consecutive time periods without liver transplantation (1985-1989, 1990-1994, 1995-1999) decreased gradually (91, 67 and 38%, respectively, with P = 0.027). This change was not observed in HBV negative cases. CONCLUSIONS: Hepatitis B virus positive FHF had a lower mortality rate than HBV negative FHF, with each group having different factors affecting mortality. PMID- 16048571 TI - Feasibility of a breath test with a substrate of natural 13C-abundance and isotope-selective non-dispersive infrared spectrometry: a preliminary study. AB - BACKGROUND AND AIMS: Isotope-selective non-dispersive infrared spectrometry (NDIRS) is a cheaper alternative to isotope ratio mass spectrometry (IRMS). We checked on the efficiency of NDIRS to perform a breath test with naturally (13)C enriched maize starch. METHODS: Fifty-six healthy volunteers were examined. After a basal fasted sample of their expiratory air was collected, the subjects ate a test breakfast of 50 g (CF50 group, n = 9) or 100 g (CF100 group, n = 47) cornflakes with 175 g yoghurt. For 6 h thereafter, exhaled air was collected every 30 min for the NDIRS measurement of (13)CO(2) concentration. RESULTS: The time-course of the curves of (13)CO(2) concentration resembled those obtained previously with IRMS. A dose-response to the amount of ingested cornflakes was observed--the maximum postprandial net increment in (13)CO(2) was statistically significantly higher in the CF100 than CF50 group: 4.78 +/- 0.13 versus 3.12 +/- 0.17 per thousand (P < 0.001), whereas the time needed to reach the maximum did not differ after the intake of 50 g (233 +/- 13 min) or 100 g (248 +/- 9 min) cornflakes. The 5-h area-under-the-curve of cumulative (13)CO(2) net recovery (AUC(60-360)) was statistically significantly greater after ingestion of 100 g compared to 50 g cornflakes (97.6 +/- 2.2 versus 61.7 +/- 2.2 micromol, P < 0.001) and it also exhibited a stable between-subject variability. CONCLUSION: The obtained proof of technical feasibility of a breath test with the use of naturally (13)C-enriched starch and NDIRS provides background for future research on the clinical usefulness of this method for a non-invasive assessment of the pancreatic exocrine function. PMID- 16048572 TI - Late complication in patients undergoing pancreatic resection with intraoperative radiation therapy: gastrointestinal bleeding with occlusion of the portal system. AB - BACKGROUND: There are few reports of late complications in patients who have undergone pancreatic resection with intraoperative radiation therapy (IORT), because carcinoma of the pancreas (PCa) and the bile duct (BCa) have a poor prognosis. The purpose of the present paper was to review gastrointestinal (GI) bleeding occurring with occlusion of the portal system (PVs) as a complication of IORT in patients surviving long term without recurrence. PATIENTS: From 1990 to 1999, 45 patients underwent surgical resection of the pancreas with IORT. Eleven of these patients survived >3 years without recurrence, and occlusion of PVs was recognized in five patients at follow-up examination. Three of these five patients received repeated blood transfusions for GI bleeding. RESULTS: One patient had BCa and two had PCa, and pancreatoduodenectomy was carried out. The delivered radiation doses of IORT were 30 Gy (two patients) and 35 Gy (one patient). The postoperative periods to initial GI bleeding were 36, 26 and 9 months, respectively. In all cases, angiography revealed occlusion of PVs and the collateral circulation. The bleeding points were esophageal varix (case 1), remnant stomach varix (case 2) and a jejunal ulcer (case 3), and blood transfusions were carried out totaling 44, 60 and 16 units, respectively. The GI bleeding disappeared spontaneously in case 1, developed sporadically in case 2 and was stopped by metallic stent insertion in PVs in case 3. CONCLUSION: During long-term follow up after pancreatectomy with IORT, it is necessary to monitor patients for GI bleeding. A clinical trial on optimum doses, long-term safety and benefit of IORT is necessary. PMID- 16048573 TI - A combination of the Helicobacter pylori stool antigen test and urea breath test is useful for clinical evaluation of eradication therapy: a multicenter study. AB - BACKGROUND: Helicobacter pylori stool antigen (HpSA) test is a new tool for evaluating the H. pylori infection. The present study was carried out to investigate the clinical usefulness of the HpSA test in the evaluation of eradication therapy by comparing it with the (13)C-urea breath test (UBT). METHODS: One hundred and five patients received eradication therapy for H. pylori. After more than 8 weeks, the success of the therapy was evaluated by the HpSA test and the UBT. Concordant results were regarded as a final diagnosis, but when the results were discordant, histological examination was carried out. RESULTS: Of the 105 patients receiving eradication therapy for H. pylori, 25 patients were regarded as H. pylori positive by the UBT and and 20 patients were regarded as H. pylori positive by the the HpSA test. Nine patients (8.6%) showed discordant results (seven cases with UBT(+) and HpSA(-), and two with UBT(-) and HpSA(+)). Five cases out of nine were ultimately judged as having a false positive result of the UBT, and in these cases the UBT values were relatively low (below 10 per thousand). The final diagnostic accuracies of the UBT and the HpSA test were 94.3% (88.0-97.9%; 95% CI) and 97.1% (91.9-99.4%), respectively. When we used the HpSA test in cases with weakly positive UBT values, we were able to diagnose the correct status of H. pylori infection after eradication in 99% of all patients (94.8-100.0%). CONCLUSION: The HpSA test is a useful tool for the evaluation of eradication therapy and a combination of the HpSA test and UBT is clinically recommended. PMID- 16048574 TI - Effects of cagA+ and cagA- strains of Helicobacter pylori on the human gastric mucus layer thickness. AB - BACKGROUND: Infection with cytotoxin-associated gene A (cagA) Helicobacter pylori is associated with severe gastric diseases, with contradictory views being expressed concerning the effect of H. pylori on the gastric mucus thickness. The aim of the present study was to differentiate between the effect of cagA+ and cagA- strains on gastric mucus thickness. METHODS: Ninety-nine patients without peptic ulcers who were not on medication were randomly recruited from consecutive endoscopy clinics: six biopsies (five antral, one body) were obtained from each patient. Cryostat sections (18 microm) were cut and stained using the modified periodic acid-Schiff/Alcian blue technique. Mucus thickness was measured using computer-assisted light microscopy. The H. pylori status was assessed by histology, Campylobacter-like organism (CLO)test and culture, and cagA+ status determined by polymerase chain reaction (PCR). RESULTS: There was no significant difference (P = 0.784) in mean mucus thickness between cagA+ (52.7 +/- 1.2 microm, n = 10), cagA- (46.6 +/- 1.1 microm, n = 18) or H. pylori-negative patients (51.3 +/- 1.1 microm, n = 30). In cagA- patients, mucus thickness was significantly reduced with increased H. pylori colonization density, Spearman (r(s)) = -0.805, P < 0.0001. In contrast, in cagA+ patients there was a weak positive, but not significant, association between mucus thickness and H. pylori colonization density, r(s) = 0.333, P = 0.381. CONCLUSIONS: The human gastric mucus thickness is not affected by infection with cagA+ or cagA- strains of H. pylori compared with uninfected. Although a trend of increased mucus thickness with cagA+ infection was observed. PMID- 16048575 TI - Diagnosis and genotyping of Helicobacter pylori by polymerase chain reaction of bacterial DNA from gastric juice. AB - BACKGROUND: Efficient and accurate detection of Helicobacter pylori infection as well as identification of virulence-associated alleles are important for the treatment of gastroduodenal diseases caused by this gastric pathogen. The present study was performed to test the efficiency of gastric juice polymerase chain reaction (PCR) method for the rapid detection of H. pylori infection and to determine the bacterial genotypes without the need for culture, which is often not feasible especially in developing countries. METHODS: DNA was extracted from gastric juice samples collected from 45 subjects and was used to amplify urease B gene (ureB) for H. pylori. Results obtained from this method were further confirmed by rapid urease test (RUT), histology and culture. Genotypes of the infected strains predicted from gastric juice PCR were compared to the genotype data obtained from the isolated strains. RESULTS: Among 45 cases, 32 were positive by RUT, 37 by histological examination, 25 by gastric juice PCR method, while culture yielded positive results for 19 samples. Except for one case, all the 19 culture-positive strains gave the same genotype with the gastric juice PCR result. It was found that the gastric juice PCR is more efficient for detection of multiple-strain infection as compared to genotype data obtained from strains isolated as pooled culture. CONCLUSIONS: This moderately sensitive technique could be employed with good efficiency, particularly in cases where it is difficult to obtain biopsy. Moreover, with this method bacterial genotype could be obtained. PMID- 16048576 TI - Changes in gastric environment with test meals affect the performance of 14C-urea breath test. AB - BACKGROUND: (14)C-urea breath test (UBT) is considered to be an accurate diagnostic test for the detection of active Helicobacter pylori infection. Various test meals are used in (14)C-UBT to slow down gastric emptying, and to enhance the gastric distribution, in order to increase the time and area of contact between microorganisms and the tracer substrate. The aim of the present paper was to evaluate the effect of gastric environment on the performance of (14)C-UBT using an alkaline and an acidic liquid test meal having gastric emptying retardant effect. METHODS: The comparison of (14)C-UBT was done with liquid test meals (200 mL water) comprising (i) plain drinking water (PDW); (ii) 1.3 g or 3.0 g citric acid (CA); and (iii) 3.0 g trisodium citrate (TSC). Eighteen patients (37 +/- 12 years, range 18-57 years) with complaints of dyspepsia participated in the study. The status of H. pylori was confirmed by histology and rapid urease test. A total of 93 kBq of (14)C-urea (0.5 mL) in a gelatin capsule was orally administered along with liquid test meals to the overnight fasting subjects. Breath samples were collected and radioactivity measured. Results were expressed as (14)CO(2)/mmol exhaled CO(2) as percentage of administered radioactive urea. RESULTS: Higher acidic gastric environment (pH approx. 2.0) with CA was found to increase the exhaled (14)CO(2) level in a dose dependent manner as compared to PDW and TSC meal (P < 0.05) at all time points. With TSC test meal, the expired (14)CO(2) level decreased in the lower acidic gastric environment (pH approx. 5.3). The peaks of exhaled (14)CO(2) with TSC test meal were observed at the same time points as that with PDW and CA test meals. The (14)C-UBT with TSC was found to be positive in 77% of patients (10/13). CONCLUSION: Better interaction between the microbial urease and (14)C urea, caused by a test meal that retards gastric emptying and that changes gastric pH, plays an important role in hydrolysis of the administered (14)C-urea by H. pylori urease. PMID- 16048577 TI - Gastric achlorhydria in H/K-ATPase-deficient (Atp4a(-/-)) mice causes severe hyperplasia, mucocystic metaplasia and upregulation of growth factors. AB - BACKGROUND: Gastric neoplasia is common in humans, yet controversy remains over contributions of chronic achlorhydria, gastrinemia and hyperplasia, to cancer risk. To study this, mice lacking the gastric H/K-ATPase (Atp4a(-/-) mice) were used to determine whether chronic loss of acid secretion, with attendant hypergastrinemia, predisposes to cancer phenotype. METHODS: Atp4a(-/-) and Atp4a(+/+) mice, paired for age and gender, were examined at 3, 8, 12 and 20 months for histopathology, and for expression of the trefoil factor family (TFF)1 3, Reg IIIbeta, gamma and delta, osteopontin, CD44, chromogranin A, Crp-ductin, and galectin, all of which are important in cell growth. RESULTS: By 8 months, the glandular stomach of the Atp4a(-/-) mice doubled in weight and thickness, and several modulators of growth were increased. Female Atp4a(-/-) mice were more hyperplastic than Atp4a(-/-) males at 12 and 20 months. By 1 year, severe mucocystic hyperplasia, incomplete intestinal metaplasia, ciliated metaplasia, a shift in mucins from neutral to acidic, and inflammation were widespread. Cells in the mucus pit zone developed a pyloric-type appearance, containing large hyaline-like, periodic acid-Schiff (PAS)-negative/alcian blue-negative inclusions. But critical characteristics of gastric neoplasia, such as nuclear atypia, invasion into the muscularis mucosa, and metastases were absent. In Atp4a(-/-) mice, chromogranin A and histidine decarboxylase, RegIIIgamma and delta, TFF3, osteopontin and CD44 were upregulated while Reg IIIbeta, and TFF1 were reduced. CONCLUSIONS: Chronic achlorhydria and hypergastrinemia in aged Atp4a(-/-) mice produced progressive hyperplasia, mucocystic and incomplete intestinal metaplasia, and the upregulation of growth factors without histological evidence of neoplasia. PMID- 16048578 TI - Detection of cytokeratin 20 mRNA in the peripheral blood of patients with colorectal cancer by immunomagnetic bead enrichment and real-time reverse transcriptase-polymeras chain reaction. AB - BACKGROUND: Detection of circulating cancer cells is a useful indicator for the risk of recurrence of advanced carcinoma. The aim of the present study was to evaluate the potential value of a novel approach to detect the circulating cancer cells in patients with colorectal cancer. This method is based on a combination of isolation of epithelial cell by a combination of negative and positive immunomagnetic beads with detection of cytokeratin 20 (CK20) mRNA by reverse transcriptase-polymeras chain reaction (RT-PCR). METHODS: Peripheral blood samples were collected from 40 patients with colorectal carcinoma on the day before operation or chemical therapy. Mononuclear cells (MNC) were isolated by centrifugation through a Ficoll gradient. Each MNC sample was equally divided into three parts and then CD45 immunomagnetic beads and/or Ber-EP4 immunomagnetic beads were used to enrich colon cancer cells. Finally, the CK20 mRNA was detected by real-time quantitative RT-PCR. As a control, LS174T colon cancer cells were serially diluted with blood from healthy individuals. RESULTS: When CD45 and Ber EP4 immunomagnetic beads were used successively, a significant correlation between CK20 mRNA levels and the initial cell concentrations was found in the control recovery experiment. The sensitivity of the assay was one cancer cell in 1 mL healthy blood. In the patient group, CK20 mRNA was detected in 80.0%, 82.5% and 72.5% of patients when CD45, Ber-EP4, and CD45/Ber-EP4 immunomagnetic beads were used, respectively. The positive detection rates of patients with colorectal carcinoma at Dukes A, B, C, and D stage were 0.0% (0/2), 33.3% (3/9), 86.7% (13/15), and 92.9% (13/14), respectively. The CK20 mRNA positive detection rate in peripheral blood was significantly correlated with tumor diameter (P < 0.01, chi(2)), lymphatic metastasis (P < 0.05) and hepatic metastasis (P < 0.05), but not with the differentiation of tumor cells. CONCLUSION: The combined use of negative and positive immunomagnetic beads followed by amplification of CK20 mRNA by means of RT-PCR is a non-invasive, sensitive, and specific assay for the detection of circulating colonic cancer cells. PMID- 16048579 TI - Apoptosis in rat jejunal mucosa is regulated partly through the central nervous system, which controls feeding behavior. AB - AIM: The aim of this study was to investigate whether central nervous system related feeding behavior regulates mucosal apoptosis in rat small intestines. METHODS: The test solutions used in this study were an H(1) receptor antagonist (chlorpheniramine maleate), 2-deoxy-D-glucose, leptin, and 1-deoxy-D-glucosamine (2-amino-1,5-anhydro-2-deoxy-D-glucitol). Test solutions were injected into the third cerebroventricles of rats. Feeding behavior and jejunal apoptosis were evaluated both with and without truncal vagotomy. Intestinal apoptosis was evaluated by percentage fragmented DNA, electrophoresis, and TUNEL staining. RESULTS: Chlorpheniramine and 2-deoxy-D-glucose elicited feeding, whereas leptin and 1-deoxy-D-glucosamine suppressed feeding. The test solutions, which elicited feeding (0.24 and 24 micromol/rat of chlorpheniramine and 2-deoxy-D-glucose, respectively), suppressed mucosal apoptosis in the rat jejunum 1 h after cerebroventricular infusion. In contrast, the test solutions, which suppressed feeding (8 and 24 micromol/rat of leptin and 1-deoxy-D-glucosamine, respectively), induced jejunal mucosal apoptosis 3 h after infusion. The effects of the test solutions on feeding behavior and changes in apoptosis were not affected by truncal vagotomy. CONCLUSION: The central nervous system, which regulates feeding behavior, might control intestinal function through the regulation of intestinal apoptosis. PMID- 16048581 TI - Images of interest. Gastrointestinal: eosinophilic esophagitis. PMID- 16048580 TI - Decreased density of interstitial cells of Cajal and neuronal cells in patients with slow-transit constipation and acquired megacolon. AB - BACKGROUND: The pathophysiology of constipation is not clearly identified as yet, and the interstital cells of Cajal (ICC), known to generate the slow wave activity and to be involved in intestinal neurotransmission and the enteric nervous system (ENS), are suspected to play an important role. The aims of the present study were to assess the distribution of ICC and neuronal cells of ENS in patients with slow-transit constipation and acquired megacolon. METHODS: Sigmoid colon specimens were obtained from patients who underwent colectomy due to slow transit constipation (n = 10), acquired megacolon (n = 9) and non-obstructive colon cancer (n = 10) as a control group. The ICC were visualized by c-Kit immunohistochemistry and neuronal cells of the ENS were demonstrated by protein gene product (PGP) 9.5. Density of cells stained by c-Kit and PGP 9.5 was calculated as percent area (area stained/area of X-Y plane) x 100, when images were collected at a magnification of x40 objective, with maximum area examined in the horizontal X-Y plane of 400 microm x 400 microm using an image analyzer. RESULTS: The densities of ICC and PGP 9.5 reactive neuronal structures were significantly decreased in all layers of sigmoid colon specimens in patients with slow-transit constipation and acquired megacolon, compared with that of the control group. However, there was no statistically significant difference in either the density of ICC or that of neuronal structures between the patients with slow-transit constipation and acquired megacolon. CONCLUSIONS: Slow-transit constipation and acquired megacolon were associated with alteration of ICC and neuronal cells of ENS in the sigmoid colon. PMID- 16048582 TI - Images of interest. Gastrointestinal: colonic intussusception in adults. PMID- 16048583 TI - Images of interest. Gastrointestinal: complications of colonoscopy. PMID- 16048584 TI - Images of interest. Hepatobiliary and pancreatic: extensive thrombosis in Budd Chiari syndrome. PMID- 16048585 TI - Images of interest. Gastrointestinal: rectal Dieulafoy lesion. PMID- 16048586 TI - Idiopathic ulcer of the small bowel containing numerous plasma cells: case resembling mucosa-associated lymphoid tissue lymphoma. AB - Idiopathic ulcer of the small bowel is a poorly recognized entity. Herein is reported a case of idiopathic ulcer of the jejunum containing numerous lymphoplasmacytoid infiltrates and mimicking mucosa-associated lymphoid tissue (MALT) lymphoma. An 82-year-old Japanese woman presented with nausea and vomiting. Following diagnosis of submucosal tumor, partial jejunal resection was performed. Macroscopically the resected specimen contained multiple small shallow ulcers. Histologically the lesion was composed of numerous plasma cells mixed with lymphocytes and histiocytes. Immunohistological study revealed that the plasma cells contained polytypic intracytoplasmic immunoglobulins. The patient remained free of disease after 36 months. Marginal zone B-cell lymphoma of MALT type arising from small intestine occasionally shows prominent plasma cell differentiation. The present case demonstrates that idiopathic ulcer of the small bowel should be added to the differential diagnosis of MALT-type lymphoma of the small bowel. PMID- 16048587 TI - CD1a immunopositivity could help to distinguish Barrett's metaplasia from heterotopic gastric mucosa. PMID- 16048588 TI - Hepatitis C virus and Behcet's disease. PMID- 16048589 TI - Characteristic endoscopic features of gastric inflammatory fibroid polyps. PMID- 16048590 TI - Human immunodeficiency virus and subtype 1B hepatitis C virus: more similarities than differences in T-helper subsets. PMID- 16048591 TI - Bone marrow dysfunction following pediatric liver transplantation. PMID- 16048592 TI - Sirolimus in pediatric solid organ transplantation. PMID- 16048593 TI - Heart transplantation: Literature review 2003-2004. AB - The rapid expansion of the volume of peer reviewed publications in recent years, including a large increase in the number of new biomedical journals, makes it difficult for the practicing clinician to stay up to date with the medical literature. This is the first review in a new series that will provide succinct summaries of the recent medical literature in the field of pediatric transplantation. In this review, we summarize important articles in the field of pediatric heart transplantation that have been published over the last 2 years (2003, 2004). The review is intended to be comprehensive but not exhaustive. PMID- 16048594 TI - Tubular and glomerular function in children after renal transplantation. AB - Glomerular and tubular function of transplanted kidneys were assessed in 46 children aged 15.7 +/- 4.6 yr, 4.2 +/- 2.8 yr after renal transplantation. There were 34 cadaveric, and 12 living-related donors. Twelve patients (26%) had acute episodes (acute tubular necrosis, rejection, or urinary tract infection) during follow-up. All patients were on triple immunosuppression. The mean serum creatinine was 1.5 +/- 0.6 mg/dL. Creatinine clearance (Ccreat) calculated from a 24-h urine collection was 48.0 +/- 19.7 mL/min/1.73 m(2), and that estimated from the Schwartz formula, 61.0 +/- 22.5 mL/min/1.73 m(2). A positive correlation was found between the calculated and estimated clearances. Mean urine concentrating ability was 487 +/- 184 mOsmol/kg, with a value lower than 400 mOsmol/kg in 35% of patients. There was a positive correlation between urine osmolality and estimated Ccreat. Metabolic acidosis (bicarbonate <22 mmol/L) was found in 41% of patients, with relatively alkaline urine and high chloride level. Fractional excretion (FE) of sodium was above 1% in 68% of patients (mean 1.66 +/- 1.06%), and FE(Mg) was above 3% (mean 10.9 +/- 5.2%) in 93% of patients. Tubular reabsorption of phosphate (TP)/glomerular filtration rate (GFR) was 3.2 +/- 0.8 mg/dL glomerular filtrate (GF). FE(K), FE(UA), and Ca/creatinine in urine were normal. There were no functional group differences between the cadaveric and living-related kidneys. Significant group differences were found in those with acute episodes and those with a normal course. Estimated Ccreat was 54 +/- 20 vs. 67 +/- 20 mL/min/1.73 m(2) in the acute episodes and the normal course groups, respectively. Also, the FE(NA), FE(UA), and FE(Mg) were higher in the acute episodes group -2.3 +/- 1.6, 10.6 +/- 4.4, and 14.8 +/- 6.5%, respectively, compared with the normal course group -1.4 +/- 0.6, 8.2 +/- 2.8, and 9.6 +/- 4.0%, respectively. There were no between-group differences in plasma bicarbonate, FE(K), TP/GFR, and urine osmolality. We believe that most, if not all tubular dysfunctions in the transplanted kidney are secondary to renal failure and interstitial damage from acute episodes and nephrotoxic drugs. These dysfunctions are similar to those in chronic renal failure, where interstitial fibrosis plays a role in kidney function deterioration. PMID- 16048595 TI - Development of growth and body mass index after pediatric renal transplantation. AB - Suboptimal final height and marked weight gain after renal transplantation (RTx) are common and may result in obesity. Steroid free immunosuppression has been advocated to improve growth and limit weight gain. We evaluated retrospectively the evolution of growth and body mass index (BMI) after renal transplantation to study risk factors for weight gain under steroid based treatment. Sixty-four pediatric patients (age 9.9 +/- 5.0 yr) were included in the study. To allow comparison between different age groups, standard deviation scores (SDS) for height and BMI for height age were calculated at time of transplantation and 3, 6, 9, 12, 24, 36, 48 and 60 months later. Induction immunosuppression consisted of basiliximab, cyclosporine and prednisone. Growth retardation at time of RTx was obvious with a SDS for height of -2.20 +/- 1.34. Height during the first year improved to an SDS of -2.0 +/- 1.27 (p < 0.01) but did further not increase in year 2 and 3. More than 40% of all patients remained 2 SDS below normal mean. SDS BMI for height age at transplantation was -0.19 +/- 0.98 and increased significantly during the first 3 months after transplantation to +0.64 +/- 1.07 (p < 0.01). Thereafter, BMI remained stable but did not decline to pretransplant values. A SDS BMI for height age of more than 2 SDS was observed in 2, 6, 9 and 11% of children at RTx and 1, 2 and 3 yr later respectively. BMI gain over 3 yr was significantly enhanced in children whose parents (especially the mother) were overweight. No influence of gender, BMI at RTx, dialysis modality prior to RTx or rejection episodes could be detected. We conclude that after RTx children exhibit some improvement in growth but height remains suboptimal. The BMI does increase significantly during the first months after RTx and does not return to baseline values under steroid-based immunosuppression. Obesity (>2 SDS above normal) does not occur more often than in the normal population. The most predictive parameter of inappropriate weight gain during 3 yr is the BMI of the mother. We would speculate that steroids may play a major role in weight gain in the early phase after RTx. However, genetic or environmental factors predict the long-term weight development. PMID- 16048596 TI - Risk factors for post-transplant lymphoproliferative disorder in pediatric patients: a case-control study. AB - Post-transplant Lymphoproliferative Disorder (PTLD) because of the Epstein-Barr Virus (EBV) is a major concern after pediatric transplantation. The group at greatest risk is EBV-seronegative recipients who receive EBV-seropositive organs. Additional risk factors remain to be determined, including those among EBV seropositive recipients. In this case-control study, PTLD cases were biopsy proven over a period of 4 yr (1997-2000, inclusive). Each case was matched with 2 controls, based on the type of organ transplanted and the period of transplantation (+/-1 yr). Variables compared between cases and controls included those relating to the clinical and virologic profiles and immunosuppressive therapy. Twenty-two cases of PTLD were diagnosed during the study period. PTLD cases occurred at a median of 22.8 months post-transplantation (range 1-131). The median age of cases was 26.2 months (range 6.1-194) compared with 47.4 months (range 0.8-202.2) for controls (p = 0.93). Cases had a higher mean baseline EBV load compared with controls (3.1 log(10) (s.d. +/- 1.0) vs. 1.6 log(10)/10(6) PBMCs (s.d. +/- 1.4), with every 1 log increase in viral load resulting in a three times increase in the likelihood of PTLD (p < 0.007). Close to one in four cases of PTLD were EBV-seropositive pretransplantation. These seropositive recipients tended to be older patients with a trend to a worse outcome compared with their seronegative counterparts. The occurrence of PTLD was not associated with the use of any specific immunosuppressants. A significant proportion of PTLD cases occurred among EBV-seropositive transplant recipients, with a tendency towards an unfavorable outcome. Besides EBV-seronegative recipients who receive seropositive organs, some EBV-seropositive pediatric patients are at risk of PTLD. Additional studies are warranted to further define the factors associated with PTLD in EBV-seropositive transplant recipients. PMID- 16048597 TI - Early discontinuation of steroids is safe and effective in pediatric kidney transplant recipients. AB - In pediatric kidney transplantation, steroid induced growth retardation and cushingoid features are of particular concern. In children, gradual steroid withdrawal late after kidney transplantation increases the risk of rejection. In this pilot study, we investigated the outcome of pediatric renal transplantation with an early steroid withdrawal protocol. This is a retrospective case-control study of pediatric renal transplants with age-matched historical control. Groups were comparable in terms of HLA matching, donor type and graft ischemia time. In the steroid withdrawal group (SWG, n = 13), induction therapy included mycophenolate mofetil (MMF) and a 5-day course of steroids with Thymoglobulin in 11 and basiliximab in two other patients. In the steroid group (SG, n = 13), in addition to steroids, four patients were given basiliximab, eight were given Thymoglobulin, and one OKT3. Maintenance therapy included tacrolimus (SWG n = 11, SG n = 3) or cyclosporine (SWG n = 2, SG n = 10). Azathioprine was given to all the patients in the SG, except the last two patients of this series who were prescribed MMF. MMF was given to all in the SWG. Patient and graft survival rates were 100% in both groups. In the SWG, no acute rejection episode was detected. In the steroid group, three patients (25%) presented with an acute rejection episode. All but one patient in either group showed immediate graft function. Patients in the steroid-withdrawal group exhibited a significantly higher creatinine clearance at 6 and 12 months post-transplant (95.8 +/- 23.3 vs. 71.3 +/- 21.9, p = 0.03; and 91.3 +/- 21.6 vs. 69.6 +/- 28.6, p = 0.04). In the SWG delta BMI was significantly lower and delta height Z score was significantly higher, and we observed significantly less hyperlipidemia, body disfigurement, and need for anti-hypertensive medication. Early steroid withdrawal in pediatric renal transplant recipients is efficacious and safe and does not increase risk of rejection, preserving optimal growth and renal function, and reducing cardiovascular risk factors. PMID- 16048598 TI - Bone mineral density in pediatric and adolescent renal transplant patients: how to evaluate. AB - Reduced bone mass is a common complication of renal transplantation in adults but only few data are present for pediatric transplant patients. Bone mineral status of pediatric renal transplant patients ages ranging from 7.5 to 17.6 years (mean age 14.9 +/- 2.3) who were at least 6 months postrenal transplantation was examined. Bone mineral density (BMD) of lumbar vertebrea and femoral neck was determined by dual energy X-ray absorptiometry (DEXA) and z-scores according to age, puberty, height and bone age were compared to sex and ethnic specific reference data. z-scores were calculated for both areal and volumetric bone density. BMD L1-4 z-scores were more than 2 SD below the mean according to chronological age in 12 patients (63%), pubertal status in six patients (31.5%), bone age in five patients (26.3%) and height in five patients (26.3%). The BMD femoral neck z-scores were more than 2 SD below the mean according to age in 10 patients (55.5%), puberty in five patients (27.7%), bone age in three (16.6%) patients and height in five (26.3%) patients. Correction of the vertebrae and femoral neck for bone size yielded osteoporotic values for seven patients (36.8%) for lumbar BMD and for four patients (22%) for femoral neck BMD. The use of aBMD in growth-retarded children has some restrictions in determining z-scores. Deficits in spinal bone density still persisted after correcting for height, puberty, bone age and volume. In renal transplant patients who have short stature it is reasonable to give values corrected for height, puberty, bone age and bone size and interpret each of these values for each patient. PMID- 16048599 TI - Infectious risk in pediatric organ transplant recipients: is it increased with the new immunosuppressive agents? AB - The risk of infection in pediatric organ transplant recipients is determined by several factors, including age, the types of organ transplanted and the immunosuppressive treatment which has dramatically changed over the past 10 yr. Little information has been reported regarding the infectious complications related to the current immunosuppressive protocols used in these children. This paper reviews (i) the immunosuppressive agents, focusing on their mechanisms of action and on the new regimens, (ii) the infections related to excessive immunosuppression and also anti-infectious properties or infectious adverse reactions associated with specific immunosuppressive agents. With the new immunosuppressive protocols, the advances in immunologic monitoring, microbiological diagnosis, anti-infectious prophylactic and preemptive treatments, strategies to minimize the risk of infection related to the immunosuppressive therapy are proposed. PMID- 16048600 TI - Post-transplantation growth among pediatric recipients of liver transplantation. AB - Improving a patient's quality-of-life (QOL) post-liver transplantation is of great importance. An aspect of improved QOL is the restoration of normal growth patterns in pediatric patients. To describe the post-transplantation growth patterns of 72 children included in the National Institute of Diabetes and Digestive and Kidney Diseases - Liver Transplantation Database (NIDDK-LTD), multilevel models were used, according to which children who waited more than a year for transplantation were smaller, compared with age and sex matched peers, at transplantation than children who waited less than a year while children who were growth retarded at transplantation experienced a larger yearly comparison height increase than children who were not growth retarded. The analysis also showed that boys older than 2 yr and younger than 13 yr at transplantation and girls older than 2 yr and younger than 11 yr at transplantation were significantly less growth retarded at transplantation than boys and girls under the age of 2 yr at transplantation. PMID- 16048601 TI - Low incidence of hepatic artery thrombosis after pediatric liver transplantation without the use of intraoperative microscope or parenteral anticoagulation. AB - The risk of hepatic artery thrombosis (HAT) after pediatric liver transplantation (PLT) has been reported to range from 0 to 25%. We report our experience focusing on the interrelationships between risk factors, surgical technique and the incidence of HAT after liver transplantation in the pediatric age group. From February 18, 1997 to December 31, 2003, 150 consecutive liver transplants were performed in 132 pediatric patients. There were similar numbers of whole grafts when compared with partial grafts, 80 (53.3%) vs. 70 (46.7%), p = 0.30. Four grafts (2.7%) developed HAT. Of the grafts with HAT, three were successfully revascularized within the first 24 h. Only one graft (0.66%) was lost to HAT. A single surgeon utilizing 3.5-6.0 magnification loupes performed all but one hepatic arterial anastomoses. All patients were followed postoperatively by a daily ultrasound protocol and with anticoagulation of aspirin and alprostadil only. Living and deceased donor left lateral segment grafts had an increased rate of HAT when compared with whole liver grafts. HAT with subsequent graft loss may be minimized in PLT with the use of surgical loupes only, anticoagulation utilizing aspirin, alprostadil, and daily ultrasounds. PMID- 16048602 TI - Heart transplantation in children: clinical outcome during the early postoperative period. AB - As improved understanding of transplant-related death should improve survival, we report a single center's experience with pediatric heart transplantation including potential risk factors and causes of death during the early postoperative period. This prospective longitudinal study involved 51 pediatric patients ranging in age from 12 days to 15.1 yr (median: 3 yr). The following pretransplant risk factors were evaluated: diagnosis, age at transplantation, recipient sex, weight and blood type, blood type match, donor/recipient sex match, weight ratio, ischemic time, recipient's status, requirement for mechanical ventilation or circulatory support, dialysis, or inotropic support at transplantation. We also determined the actuarial survival, clinical outcomes, and causes of death in this population. Survival was 86% during the early postoperative period (200 mg/dL were observed in 33 children. Thrombocytopenia (platelet count <140 000) was not observed. We concluded that early outcomes with sirolimus were acceptable with 98% graft survival and 11% incidence of acute rejection. Medication was discontinued in 20% for adverse events which included poor wound healing and non-infectious pneumonitis. Infections with cytomegalovirus and Epstein-Barr virus, and PTLD were also significant early complications. Therefore, a sirolimus-based regimen that is combined with both an interleukin-2 receptor antibody and a calcineurin inhibitor may be excessive immunosuppression for pediatric renal transplant recipients. PMID- 16048607 TI - Effect of age on lipid profiles in pediatric heart transplant recipients. AB - This study's objectives were to determine if pediatric orthotopic heart transplant (OHT) recipients over all ages develop hyperlipidemia and, secondarily, to identify the effects of immunosuppressive agents and statins on lipid profiles in these patients. Retrospective chart review was performed for pediatric patients transplanted between January 1987 and June 2002. Of the 100 OHTs performed, 50 patients satisfied the inclusion criteria and were grouped by age at OHT as follows: group 1 (n = 16): 0-4 yr; group 2 (n = 10): 5-9 yr; group 3 (n = 15): 10-14 yr; group 4 (n = 9): 15-18 yr. There were 2789 lipid levels recorded, and each patient had an average of 14 post-OHT lipoprotein panels measured. Post-OHT total cholesterol and low-density lipoprotein (LDL) levels were significantly greater than those of the general population for the entire follow-up period in all age groups, except for LDL levels in group 2. Cyclosporin level and prednisone dose were positively associated with total cholesterol and LDL levels (p < 0.03). Statins significantly decreased total cholesterol and LDL levels (p < 0.001). Hyperlipidemia affects OHT patients of all ages. Even the youngest patients may benefit from immunosuppression using an alternative to cyclosporin, such as tacrolimus, and steroid-free regimens, which may improve lipid profiles. Once safety and efficacy data are available, all age groups may benefit from statins. PMID- 16048608 TI - Duct-to-duct biliary reconstruction in pediatric living donor liver transplantation. AB - The results of duct-to-duct biliary reconstruction in six pediatric patients who received a living donor liver transplant aged from 2 months to 11 yr old are reported. The graft was either entire or a part of the left lateral segments. The orifice of the bile duct of the graft was anastomosed to the recipients' hepatic duct in an end-to-end fashion by interrupted suture using 6-0 absorbable material. A transanastomotic external stent tube (4 Fr) was passed through the stump of the recipients' cystic duct. Mean time for reconstruction was 24 min. All the recipients survived the operation and reinitiated oral intake on postoperative day 3. There were no early biliary complications. One 5-yr-old boy suffered from an anastomotic stenosis 9 months after transplantation. He underwent re-anastomosis by Roux-en Y (R-Y) procedure and recovered uneventfully. Duct-to-duct anastomosis in pediatric living donor liver transplantation has benefits while the complication rate is comparable to R-Y reconstruction. PMID- 16048609 TI - Successful bone marrow transplantation for life threatening xanthogranuloma disseminatum in neurofibromatosis type-1. AB - A 2-yr and 9-month-old female patient with neurofibromatosis type-1 presented with hepatomegaly, anemia, thrombocytopenia, and croupy cough and diagnosed with xanthogranuloma disseminatum (XD). She failed chemotherapy consisting of steroids, 6-mercaptopurine and methotrexate. A partial response to HLH-94 therapy that included etoposide and cyclosporine A was initially observed. However, she continued to have significant organ dysfunction without further improvement at 6 months of therapy. She then received matched unrelated donor bone marrow transplantation (BMT) following carmustine, etoposide, cytarabine and melphelan conditioning with complete resolution of symptoms. BMT is an option in therapy resistant, life threatening XD cases. PMID- 16048610 TI - Successful recovery of aplastic anemia following orthotopic liver transplantation for non-A-E acute liver failure. AB - This report describes a teenager who developed aplastic anemia (AA) because of non-A-E acute liver failure (ALF) requiring orthotopic liver transplantation (OLT). His AA did not recover spontaneously and he required treatment with ATG 9 months post-OLT. Bone marrow recovery occurred 4 months after immunotherapy and coincided with further intensification of immunusuppression required to treat early chronic rejection of the liver graft. Three years post-OLT he remains well with good bone marrow and liver function. Intensification of immunosuppression can lead to successful resolution of AA associated with non-A-E ALF. PMID- 16048611 TI - Cardiac tamponade in a pediatric renal transplant recipient on sirolimus therapy. AB - Because of its lack of nephrotoxicity, the use of sirolimus, as an immunosuppressive agent, has increased considerably in solid-organ transplant (Tx) recipients. With its increased use, Tx professionals are encountering a variety of previously unreported side-effects such as angioedema and interstitial pneumonitis. We describe here the case of a pediatric renal Tx recipient who, while receiving sirolimus, developed a large pericardial effusion requiring pericardiocentesis. An extensive workup for an infectious etiology was performed; the only positive result was isolation of adenovirus type 2 from the patient's stool specimen. Following sirolimus dose reduction this child's effusion stabilized and has not recurred. The purpose of this report is to advise health care professionals caring for Tx recipients about this potentially life threatening complication associated with sirolimus. The role of adenovirus, if any, in contributing to the development of our patient's pericardial effusion is discussed herein. PMID- 16048612 TI - Idiopathic intracranial hypertension following kidney transplantation: a case report and review of the literature. AB - A pediatric kidney transplant recipient receiving tacrolimus for immunosuppression experienced symptoms consistent with idiopathic intracranial hypertension. The diagnosis of idiopathic intracranial hypertension and possible secondary causes of intracranial hypertension are reviewed in association with the patient's clinical course. Treatment options for the reversal of intracranial hypertension are summarized. Because of the complexity of associated conditions in kidney transplant recipients, symptoms of persistent headaches, visual changes and nausea and vomiting should be promptly investigated by fundoscopic examination in the setting of immunosuppression therapy to prevent vision loss. PMID- 16048614 TI - Think global, act regional: promoting change in medical education. PMID- 16048615 TI - Looking for the child's perspective. PMID- 16048618 TI - The nature of qualitative comments in evaluating professionalism. AB - INTRODUCTION: We analysed written comments on clerkship evaluation forms to determine if they furthered our understanding of students' professional behaviour. METHODS: We transcribed all written comments related to professional behaviour for 1 medical school class in 4 major clerkships, then categorised the comments. Comments were coded into categories by two raters. They were also coded as positive, negative or equivocal. The 3 types of comments were each correlated with a Likert-type score for professionalism for each student in these same clerkships. RESULTS: Most comments covered initiative, working well with others, patient skills and working hard. Of the 1845 comments, 1721 were positive, 106 were negative and 18 were equivocal, neither wholly positive nor wholly negative. The total number of positive comments correlated with the students' numerical professionalism score (r = 0.29, P < 0.001); negative comments correlated negatively with the numerical professionalism score (r = -0.44, P < 0.001). Equivocal comments were significantly negative (r = -0.25, P = 0.002). Qualitative analysis revealed that equivocal comments were focused on maturity, initiative and teamwork. CONCLUSIONS: Written comments contain unique indicators of students' professional behaviour and are largely positive in nature; negative comments stand out by their rarity and intensity. There is a set of negative comments masked by evaluators in language that appears neutral or even partly positive that statistically resemble negative comments and should be regarded as such by students who receive this type of comments. Evaluators should be encouraged and trained to express their qualitative views of students. PMID- 16048619 TI - Defining professionalism in anaesthesiology. AB - BACKGROUND: The Royal College of Physicians and Surgeons of Canada, through the CanMEDS 2000 project has identified the role of professional as 1 of 7 roles for which physicians are expected to be accountable when dealing with patients. Each specialty is responsible for defining this role relative to their specialty. METHODS: The qualities of professionalism for anaesthesiology were defined through a Delphi process involving Canadian anaesthesiology educators. The panellists took part in 3 rounds that identified qualities in 3 distinct areas of professionalism - humanistic qualities, personal development qualities and anaesthesiology meta-competences. RESULTS: A total of 23 of 29 anaesthesiologists responded (79%) in round 1, with response rates to rounds 2 and 3 being 72% and 69%, respectively. Of the original 36 qualities, some were combined, definitions were changed in 23, 11 qualities were added and 4 were deleted, leaving a list of 40 qualities. DISCUSSION: There is considerable interest in this issue among the Canadian educators in postgraduate anaesthesiology. Consensus on important professional qualities for anaesthesiologists was obtained through the Delphi technique. These qualities will form the basis of identifiable professional behaviours to which anaesthesiologists should aspire. PMID- 16048620 TI - Everyday classism in medical school: experiencing marginality and resistance. AB - OBJECTIVE: To explore the medical school experiences of students who self identify as coming from a working-class or impoverished family background. METHODS: A questionnaire was administered to Year 3 medical students at a Canadian medical school and in-depth interviews were held with 25 of these students (cohort 1). The same methods were repeated with another Year 3 class 3 years later (cohort 2). RESULTS: While having (or not having) money was the most obvious impact of social class differences, students also discussed more subtle signs of class that made it easier or more difficult to fit in at medical school. Students from working-class or impoverished backgrounds were significantly less likely to report that they fitted in well, and more likely to report that their class background had a negative impact in school. They were also more likely to indicate awareness that a patient's social class may affect their health care treatment. CONCLUSION: Students from working-class or impoverished backgrounds may experience alienation in medical school. Through the commonplace interactions of 'everyday classism' they may experience marginalisation, isolation, disrespect and unintentional slights. At the same time, they suggest that their experiences of exclusion may strengthen their clinical practice. PMID- 16048621 TI - Explanations in consultations: the combined effectiveness of doctors' and nurses' communication with patients. AB - BACKGROUND: Multidisciplinary and interprofessional working is currently a priority in health care policy, in caring for patients and in health professional education. Realising multidisciplinary approaches presents challenges in the context of changes in doctors' and nurses' roles and the increased emphasis placed on communication with patients. In communication in consultations, explanations are employed in the service of numerous activities, including decision making, diagnosis and physical examination, but they have been little studied. SETTING: This paper presents findings from a comparative study of doctors' and nurses' communication with patients in multidisciplinary health care, focusing on diabetes in primary care. METHODS: Video- and audio-recorded consultations were subjected to conversation analysis. Output from discussion groups with patient representatives and health professionals underwent qualitative analysis. FINDINGS: Distinctive features of explanations in nurses' and doctors' consultations with patients were identified. These can be understood by reference to patterns of communication. Nurses' communication was mediated by patients' contributions; doctors' communication gave an overarching direction to the consultation as a whole. While nurses' explanations began from the viewpoint of a patient's responsibility and behaviour, doctors' explanations began from the viewpoint of biomedical intervention. Their consultations lent different opportunities for patients' involvement. CONCLUSION: Nurses' and doctors' communications each exhibit their own distinct features. Specification of these features, when considered in the context of a particular consultation activity such as explanations, allows both recognition of the distinct contributions each profession can offer and identification of ways of combining these to maximum effect. This has implications for policy, for practice and for interprofessional education. PMID- 16048622 TI - Teaching paediatric residents about learning disorders: use of standardised case discussion versus multimedia computer tutorial. AB - BACKGROUND: We developed a standardised case-based educational exercise on the topic of childhood learning disorders, and a multimedia computerised adaptation of this exercise, as part of a national curriculum project based on the Bright Futures guidelines. OBJECTIVE: To explore resident perceptions of the facilitated case discussion (FCD) and the computerised tutorial (CT). DESIGN: Quasi randomised comparison of two educational interventions. SETTING: Preclinic teaching conferences at a large urban children's hospital. PARTICIPANTS: A total of 46 paediatric residents years 1-3 assigned to either FCD (n = 21) or CT (n = 25). INTERVENTIONS: FCD residents met in groups of 8-12 with a trained facilitator for a structured case discussion, while CT residents worked in groups of 2-3 at a computer station linked to an interactive website. OUTCOME MEASURES: Participant responses during semistructured focus group interviews. ANALYSIS: Focus group transcripts, field notes and computer logs were analysed simultaneously using qualitative grounded theory methodology. RESULTS: Residents experienced CT as fun, offering flexibility, greater auditory and visual appeal and more opportunities for active learning. FCD allowed greater contact with expert faculty and made the material more relevant to clinical practice. FCD participants emphasised the clinical skills gleaned and stated that the learning experience would change their future patient management. Both groups reported that case discussion was more interactive than computer learning. Median time spent on learning was slightly shorter for the CT group. All groups of learners arrived at the correct final diagnosis. CONCLUSIONS: FCD and CT stimulate different types of learning among paediatric residents. Future studies are needed to determine how to integrate these two techniques to meet the learning needs of residents in diverse settings. PMID- 16048623 TI - Doctors' consultations with children and their parents: a model of competencies, outcomes and confounding influences. AB - CONTEXT: The clinical consultation is an important aspect of the doctor's role. However, there is a particular shortage of methods for assessing its quality, and its complexity makes it a considerable assessment challenge. RESEARCH QUESTION: What are the key components of consultations involving children? METHODS: (1) A content analysis of relevant published and unpublished literature. (2) A nominal group consensus exercise with experienced paediatricians. RESULTS: The content analysis and consensus exercise suggested similar lists of doctor's characteristics, tasks and outcomes as being important components of the consultation. Doctor's characteristics include: clinical judgement, clinical knowledge, physical examination, information gathering, clinical questioning, information giving, patient-centredness, parent-centredness, interpersonal skills, and consultation management. Important tasks include: organisation and efficiency, rapport, information gathering, getting the family perspective, examination and procedures, evaluation, medically appropriate plans, family appropriate plans, enhancing understanding and recall, achieving consensus, sharing responsibility, family knows how to get further help and liaison with other relevant health-care professionals. Important outcomes include: family satisfaction, family perceptions, compliance, health, health-related problems and doctor's satisfaction. The studies reviewed in the literature also provided a catalogue of factors that have been shown to influence the doctor-patient interaction that could potentially confound the assessment of a doctor's performance. These include the doctor's: age, gender, training, speciality, income, social class and politics; the patient's: age, gender, health, prognosis, social class, education, health beliefs and preferences about control and risk. The length of the acquaintance between doctor and patient, and the workload and case-mix in the clinic also affect the interaction. In several studies it is clear that particular combinations of doctor-type and patient-type have especially good or bad interactions. CONCLUSIONS AND FURTHER WORK: These components are synthesised in a single model of the doctor-patient interaction to guide the development and evaluation of assessment instruments aimed at consultations involving children. PMID- 16048624 TI - Children and their parents assessing the doctor-patient interaction: a rating system for doctors' communication skills. AB - CONTEXT: Only a patient and his or her family can judge many of the most important aspects of the doctor-patient interaction. This study evaluates the feasibility and reliability of children and their families assessing the quality of paediatricians' interactions using a rating instrument developed specifically for this purpose. METHODS: A reliability analysis using generalisability theory on the ratings from 352 doctor-patient interactions across different speciality clinics. RESULTS: Ratings were normally distributed. They were highest for 'overall' performance, and lowest for giving time to discuss the families' agenda. An appropriate sample of adults' ratings provided a reliable score (G = 0.7 with 15 raters), but children's ratings were too idiosyncratic to be reproducible (G = 0.36 with 15 raters). CONCLUSIONS AND FURTHER WORK: Accompanying adults can provide reliable ratings of doctors' interactions with children. Because an adult is usually present at the consultation their ratings provide a highly feasible and authentic approach. Sampling doctors' interactions from different clinics and with patients of both genders provides a universal picture of performance. The method is ideal to measure performance for in training assessment or revalidation. Further work is in progress to evaluate the educational impact of feeding ratings back to the doctors being assessed, and their use in a range of clinical contexts. PMID- 16048625 TI - Medical education in Southeast Asia: emerging issues, challenges and opportunities. AB - CONTEXT: Medical education in Southeast Asian countries is undergoing rapid changes, with the realignment of medical schools' curricula to meet national needs and priorities, the adoption of and experimentation with innovations, and greater emphasis on staff development initiatives. The Medical Education Unit of the National University of Singapore undertook a project to compile the educational processes taking place in these medical schools. METHOD: This was a process-focused initiative. We developed a custom-made questionnaire that emphasised narrative description. We compiled profiles of 30 selected medical schools in the Southeast Asian region and, after editing and review, published these profiles. This report summarises the key findings from the project. FINDINGS: Medical schools in Southeast Asia are in a dynamic state. Schools are gradually adopting student-centred learning approaches, including problem-based learning. Many schools offer their students early clinical training, opportunities for out-of-hospital postings, and student-selected electives. Multiple-choice questions and oral examinations are almost universally used in student assessment. Portfolios and self- and peer-assessments are seldom used. Major challenges faced by the schools involve shortages of qualified staff and financial constraints. Major goals for the future include the implementation of student-centred learning, the revamping of the assessment process, and staff training. CONCLUSIONS: This is a compilation of self-reported profiles of selected medical schools where we encouraged self-reflection and analysis. As with other surveys, there could be potential self-selection bias. Nevertheless, the profiles of these selected schools provide a broad overview of the status of medical schools in Southeast Asia. PMID- 16048626 TI - Analysis and prospects for curricular reform of medical schools in Southeast Europe. AB - AIM: To analyse the curricula of 16 medical schools in 6 countries in Southeast Europe in order to establish a prevailing standard curriculum against which a prospective curriculum reform could formulate its objectives. METHODS: Curricular information was gathered from a questionnaire sent via e-mail to the respective medical schools. The data collected ranged from the numbers of enrolled students to a breakdown of courses with distribution of instruction hours for certain teaching formats. For easier comparison the courses were clustered into 5 groups: pre-clinical, clinical, public health, liberal arts and electives. RESULTS: Belgrade has the highest number of undergraduate students, while Mostar has the lowest. Novi Sad, Foca/Srbinje, Zagreb, Split, Sofia, Ljubljana and Mostar have more than 5000 instruction hours, but Sarajevo lags behind with 4005 hours. Anatomy dominates the course load in Year 1, ranging from a share of 18.4% in Sofia to 11.3% in Novi Sad. Physiology dominates Year 2, ranging from 16.8% in Rijeka to 8.9% in Split, whereas in Year 3 the dominating course is pathology, reaching a peak of 13.7% in Sarajevo. Sofia has the highest number of class hours of clinical courses. The predominant public health courses are social medicine, family medicine and medical ecology. Medical English is taught at all medical schools (as electives in Ljubljana and Rijeka). CONCLUSION: There is considerable potential for curriculum improvement in the region. Teacher training, student participation, the definition of core competencies and the introduction of new methodologies should all be implemented in the process. PMID- 16048627 TI - Developing an education and assessment framework for the Foundation Programme. AB - AIM: To develop an education and assessment framework for the second year of the Foundation Programme (F2). METHODS: A total of 23 PRHOs were recruited to the F2 pilot in August 2003. The training posts included a variety of specialties at 2 hospital trusts plus primary care. Trainee expectations and satisfaction were evaluated using questionnaires administered before and at the end of the pilot. At the end of the pilot, 10 trainees participated in a focus group and 19 trainers participated in a semistructured telephone interview. RESULTS: The majority of trainees (78%) felt that their expectations of the F2 pilot were met and all felt that they had improved their generic skills. Attendance at the generic education programme was 95%. The majority of trainees found the assessment framework useful. The percentage of undecided trainees in terms of career aspirations dropped from 48% to 13%. Trainees valued the breadth of experience provide by the year and the support provided by the programme directors and each other. A need for better communication, administrative support and time for assessment was highlighted by the trainers. CONCLUSIONS: Early, focused education on generic skills will benefit both doctors and their patients. More varied career experience will help to ensure that doctors make appropriate and timely career decisions. Pilots are identifying good practice and areas that need improvement. PMID- 16048628 TI - Effects of item and rater characteristics on checklist recording: what should we look for? AB - OBJECTIVE: Examinations based on using standardised patients (SPs) commonly use checklist recordings to evaluate students' clinical performance. This paper examines whether and to what extent item and rater characteristics affect the reliability of history checklist recording in an SP-based assessment. METHODS: Checklist items were reviewed for the presence or absence of 5 item characteristics and a 2-point versus 3-point scoring scale. Agreement between checklist recordings obtained from SPs and clinician-examiners (CEs) were compared by item characteristics, scoring scale and CEs' level of involvement in the assessment. RESULTS: Based on 3179 pairs of recordings, the overall percentage of agreement between SPs and CEs was 83% (kappa = 0.64). Agreement was significantly higher for items scored on a 2-point than on a 3-point scale, and when the CE was also the author and the trainer of the station. After controlling for other factors, item characteristics were only marginally associated with level of interrater agreement. CONCLUSIONS: This study suggests that attention should be paid to specific aspects of checklist development and checklist recording training when an SP or CE is used as recorder. PMID- 16048629 TI - Informal learning in postgraduate medical education: from cognitivism to 'culturism'. AB - BACKGROUND: Work-based learning occupies a central role in the training and ongoing development of the medical workforce. With this arises the need to understand the processes involved, particularly those relating to informal learning. Approaches to informal learning in postgraduate medical education have tended to consider the mind as an independent processor of information. METHOD: In this paper, such cognitive approaches are critiqued and an alternative socio cultural view on informal learning described. Recent and imminent changes in postgraduate medical education are identified, namely the reduction in patient experience, the fragmentation of teaching, and the development of competency frameworks and structured curricula. It is argued that although the latter may be useful in the construction of formal learning programmes, they will do little to enhance the progression of the individual from newcomer to old-timer or the cultural assimilation of the learner into a profession. DISCUSSION: Strategies for enhancing informal learning in the workplace are recommended in which increased attention is paid to the development of the medical apprentice within a community of social practice. These include the establishment of strong goals, the use of improvised learning practices, attention to levels of individual engagement and workplace affordances, immersion in professional discourse and behaviours, support in relation to the development of a professional identity and the provision of opportunities to transform social practice. PMID- 16048630 TI - The chromatoid body in spermatogenesis. AB - All germ cells throughout the animal kingdom contain cytoplasmic cloud-like accumulations of material called nuage. Polar bodies in Drosophila oocytes are probably the best known forms of nuage. In spermatogenic cells, the nuage is called chromatoid body (CB). In early spermatids of the rat, it has a diameter of 1-1.5 microm and a finely filamentous lobular structure. Typically, it is associated with a multitude of vesicles. It is first clearly seen in mid- and late pachytene spermatocytes as an intermitochondrial dense material. During early spermiogenesis it is seen near the Golgi complex and frequently connected by material continuities through nuclear pore complexes with intranuclear particles. In living cells, the CB moves around the Golgi complex and has frequent contacts with it. The CB also moves perpendicularly to the nuclear envelope, and even through cytoplasmic bridges to the neighbour spermatids. One of the major components of the CB is a DEAD-box RNA helicase VASA that belongs to a class of proteins thought to act as RNA chaperones. It is a general marker of all germ cells and best characterized in Drosophila. The mouse VASA homologue was recently used as a marker of sperm formation from embryonic stem cells. It becomes generally accepted that the CB with its associated structures constitute a mechanism of post-transcriptional processing and storage of several mRNA species that are shared between neighbour cells and used for translation when the genome of the spermatids becomes inactive. PMID- 16048631 TI - Intracytoplasmic sperm injection as a complement to gonadotrophin treatment in infertile men with hypogonadotrophic hypogonadism. AB - In this study we sought to determine whether intracytoplasmic sperm injection (ICSI) could improve the efficacy of treatment with gonadotrophins in gonadotrophin-deficient men in terms of pregnancy. A series of six adult men (aged 26-47 years) with hypogonadotrophic hypogonadism (HH) is reported: four men with prepubertal isolated idiopathic HH (IIHH) and two adult-onset HH, as part of hypopituitarism secondary to surgical treatment of a pituitary tumour. All were azoospermic. To restore spermatogenesis, all received hormonal treatment with intramuscular human menopausal gonadotrophins (HMG) and human chorionic gonadotrophin (HCG) for 2 to 23 months. High basal serum inhibin B was predictive of rapid and complete recovery of spermatogenesis. In the two adult-onset HH, a natural pregnancy was achieved within 3 months. The four men with IIHH underwent ICSI because of poor sperm quality. ICSI using fresh or frozen-thawed ejaculated spermatozoa was performed after 6-23 months of gonadotrophin treatment. ICSI provided good clinical results in terms of fertilization and embryo quality, and resulted in three pregnancies that ended in three term deliveries. In men with oligozoospermia related to prepubertal IIHH, ICSI shortens the hormonal treatment and enhances the chances of pregnancy. PMID- 16048632 TI - Sperm concentration in Latvian military conscripts as compared with other countries in the Nordic-Baltic area. AB - Recent studies of semen quality in men from the general population gave rise to the hypothesis of an East-West gradient in semen quality in the Nordic-Baltic area, with the highest sperm counts in Estonia, Lithuania and Finland, and the lowest in Denmark (30% difference in mean concentration). Genetic, lifestyle related and environmental factors - alone or in combination - were suggested to be responsible for these differences. The aim of this study was to assess sperm concentration in men from the general population in Latvia and to investigate the impact of ethnic and lifestyle-related factors on this marker of male reproductive health. A total of 133 military conscripts from Latvia were investigated. We found that sperm counts among Latvian adolescents were at the same level (mean 74, median 63 x 10(6)/mL) as those previously reported from Estonia, Lithuania and Finland. Sperm concentration was somewhat higher than in Sweden without reaching the level of statistical significance (mean difference 3 x 10(6)/mL; 95% CI: -10, 16 x 10(6)/mL), and statistically significantly higher that in Denmark (mean difference: 17 x 10(6)/mL; 95% CI: 5, 2 x 10(6)/mL). The study also revealed an impact of ethnic factors on sperm numbers. Sperm concentration was significantly higher in men with both parents born in Latvia (77 +/- 60 x 10(6)/mL), compared with men with both parents born outside Latvia (55 +/- 45 x 10(6)/mL, p = 0.03). PMID- 16048633 TI - CREM activator and repressor isoform expression in human male germ cells. AB - The transcription factor cAMP-responsive element modulator (CREM) is known to play a vital role for male fertility as it has been demonstrated that male mice lacking a functional CREM gene are infertile. The CREM gene consists of 14 exons. Owing to alternative exon splicing, CREM gene expression results in the production of functionally different CREM proteins with either activating or repressing potential on target gene expression. Infertile men have been reported to reveal a substantial reduction of CREM activators and additional inaccurately spliced CREM transcripts. In the present study, we analysed the expression of CREM transcripts with the recently reported leader exons theta1 and theta2 and identified a new putative CREM repressor, namely theta1-F-H, in patients with impaired spermatogenesis. In addition, we applied single cell microdissection followed by RT-PCR with leader exons B, theta1 and theta2 to assign the expressed CREM activator and repressor isoforms to specific germ cell types within the seminiferous epithelium. Contrary to dogma, we demonstrated CREM activator and repressor isoforms in all germ cell types, but not in Sertoli cells. However, the percentage of germ cell samples that revealed positive RT-PCR signals for these CREM activators was higher in spermatocytes and round spermatids than in spermatogonia and elongated spermatids. It remains unknown whether these activator transcripts are physiologically active. Our data suggest a fine-tuning between CREM activator and repressor isoforms in normal germ cells that might be disturbed during impaired spermatogenesis. PMID- 16048634 TI - Quantification of survivin mRNA in testes of infertile patients and in testicular germ cell tumours: high levels of expression associated with normal spermatogenesis. AB - Deregulated apoptosis of germ cells may contribute to male infertility as well as malignant transformation. Survivin, an inhibitor of apoptosis (IAP), is overexpressed in all the most common human malignancies, but barely detectable in normal tissues. We used real-time polymerase chain reaction (PCR) to quantify survivin mRNA expression in normal testes (n = 22), testes with defective spermatogenesis (n = 26) and testicular germ cell tumours (TGCTs; n = 16). Survivin was expressed at high levels in normal testes. Testicular survivin levels in infertile patients were related inversely to the severity of spermatogenic failure (p < 0.001), with a lack of expression in most specimens with pre-meiotic spermatogenic arrest and in all those with germ cell aplasia. Lower levels of expression were observed in TGCTs than in normal testes. While survivin expression was detected in most TGCTs with undifferentiated components (12 of 13), it was absent in all mature teratomas (n = 3). These data show that survivin is expressed in normal and transformed germ cells. Its downregulation in spermatogenic disorders indicates that survivin may contribute to the normal balance between germ cell proliferation and apoptosis. In TGCTs, survivin expression appears to be lost with somatic differentiation. PMID- 16048635 TI - Sites of residual spermatozoa after irrigation of the distal vas deferens using normal saline solution during vasectomy in a rat model. AB - We have previously reported that irrigation of the distal vas deferens using a normal saline solution (NSS) is successful in removing a large amount of spermatozoa from the tract. However, this technique does not completely remove all the motile spermatozoa from the ejaculate. The aim of the present study is to evaluate the location of the residual spermatozoa after distally irrigating the vas deferens. Twenty male Sprague-Dawley rats (400-450 g) constitute our study population. The animals were divided into two groups: group 1, control group (n = 10), rats that undergo only vasectomy and group 2, experimented group (n = 10), rats that undergo vasectomy and distal irrigation of the vas deferens using 3 mL of NSS. In both groups, the middle and terminal parts of the vas deferens including the seminal vesicles are removed and sent for spermatozoa count. The post-vasectomy urine samples containing spermatozoa are obtained by mid-ventral cystocentesis and the concentration is determined using a haemocytometer. More spermatozoa was found in the urine samples of the experimented group than the control (21.3 +/- 10.61 vs. 0.2 +/- 0.20 million/ml, p-value = 0.068), and lesser residual sperms reside at both the middle and the terminal parts of the vas deferens (0.5 +/- 0.31 vs. 3.0 +/- 0.00; p = 0.008 and 1.1 +/- 0.99 vs. 2.0 +/- 0.00; p = 0.036 respectively). No sperms were present in the seminal vesicles of the control group, but two of 10 rats in the experimented group had few to moderate amount of sperms in their seminal vesicles (p = 0.180). After the distal irrigation of the vas deferens using NSS, some residual sperms resided in the middle and more at the distal part of the vas with a few that escaped into the seminal vesicles. PMID- 16048636 TI - Altered melatonin secretion in hypogonadal men: clinical evidence. AB - The pineal gland, through the rhythmic production of melatonin, seems to play an important role in the control of the reproductive function of many vertebrate species. In contrast, the effects of the pineal gland in humans and the relationship between gonadotropins and melatonin secretion are not yet clarified. On the basis of these considerations, the aim of the present study was to clarify whether melatonin serum concentrations were altered in males with different hypothalamo-pituitary-gonadal disturbances, in comparison to normal individuals. We have studied 36 individuals divided into three groups according to their gonadotropin status: normals, hypogonadotropic hypogonadism and hypergonadotropic hypogonadism. They were submitted to blood sample withdrawal at 03.00, 11.00 and 19.00 h for melatonin determination according to a radioimmunological method, without extraction of the sample. The results obtained in the present study suggest the existence of an interaction between the pituitary and the pineal gland. In fact, in the case of hypersecretion of gonadotropins, nocturnal melatonin release is reduced, while night melatonin secretion is increased in the opposite situation (hypogonadotropic hypogonadism). Both these endocrine pathologies are characterized by a reduced sexual steroid secretion; for that reason, this reduction cannot be regarded as responsible for the two opposite dysfunctions of melatonin release. In conclusion, our study shows that darkness dependent release of melatonin in males with hypogonadotropic hypogonadism is significantly higher in comparison with the healthy men, while it is significantly reduced in patients with hypergonadotropic hypogonadism. A strong significant negative correlation is also found between gonadotropins and melatonin release. PMID- 16048637 TI - Human mtDNA haplogroups and reduced male fertility: real association or hidden population substructuring. AB - A mitochondrial DNA (mtDNA) haplogroup association study carried out in 101 southern Portuguese males with oligozoospermia showed to be negative when comparing with a geographically matching control sample. Misleading positive association signs were however obtained when using other control samples from the same country. This shows that mtDNA population substructure can also introduce spurious signs in haplogroup association studies, as previously reported for Y chromosome. However, our data do not exclude the probability that a particular mtDNA mutation contributes significantly to the reduction of sperm production, but indeed precludes the hypothesis of a significant association between such a mutation and specific haplogroup. PMID- 16048638 TI - The cryptorchidism prevalence among infants in the general population of Rotterdam, the Netherlands. AB - Published trends and geographical differences in cryptorchidism rates are almost exclusively derived from hospital-based birth defect registers, which are sensitive to selection bias and incomplete reporting. This study aimed to accurately assess the cryptorchidism prevalence in the general population of Rotterdam. Of 7652 consecutive male live births, 7292 (95%) were examined for cryptorchidism at Child Healthcare Centres around the age of 1 month. In a subgroup of cases, the persistence of cryptorchidism was re-assessed during a follow-up examination by expert specialists. The cryptorchidism rate at the median age of 35 days was 1.2% (89/7292). In the re-examined subgroup (median age 95 days) 69% of the boys (24/35) had persistent cryptorchidism, of which 20 were unilateral and four bilateral. The population rate of 1.2% falls within the range of 0.9-9% reported by others. Differences in case ascertainment and population characteristics probably explain part of the differences between studies. Our cross-sectional design does not allow for analysis of a temporal trend, but provides a baseline for future trend studies. To study cryptorchidism rates, trends, and risk factors, a systematic case ascertainment is warranted. PMID- 16048639 TI - Deletion of LTbetaR augments male susceptibility to Plasmodium chabaudi. AB - SUMMARY Disruption of the lymphotoxin beta receptor (LTbetaR) gene has been shown to result in enhanced resistance of female mice to blood-stage Plasmodium chabaudi malaria. Here, we investigate the effect of LTbetaR deletion on host defence of males. In contrast to females, male LTbetaR(-/-) mice do not exhibit any increase in resistance. Conversely, they are even more susceptible than wild type C57BL/6 mice, which becomes evident after lowering circulating levels of testosterone by castration, which makes C57BL/6 males resistant, whereas LTbetaR( /-) remain susceptible. Gene-expression analysis using cDNA arrays revealed no differences in immunological responses in spleen of malaria-resistant female and malaria-susceptible castrated male LTbetaR(-/-) mice. In the liver, however, expression levels of plasminogen activator inhibitor PAI1, chemokine CXCL10, dual specificity phosphatase DUSP1, and hydroxysteroid-specific sulfotransferases Sult2a1/2 were decreased 6- to 85-fold in susceptible castrated male LTbetaR(-/-) mice in comparison to resistant female LTbetaR(-/-) mice at maximal parasitaemia, as evidenced by Northern blot analyses. The present data support our previous view that the liver is involved in the combat against malarial blood stages and that down-regulation of the genes DUSP1 and Sult2a1/2 signals dysregulation of protective liver responses, thus possibly contributing to male susceptibility of LTbetaR(-/-) mice. PMID- 16048640 TI - Genetic resistance to Teladorsagia circumcincta: IgA and parameters at slaughter in Churra sheep. AB - SUMMARY Previous experiments have shown that genetic resistance to infection by Teladorsagia circumcincta in sheep can be measured by the level of IgA in gastric mucus, jointly with other parameters. The aim of this study has been to observe the influence of IgA on adult worms. The experiment was carried out with Churra sheep experimentally infected with T. circumcincta. At slaughter, gastric content, gastric mucus, blood samples and faeces were recovered to determine the number of eggs in utero, length of adult females, worm burden, number of L4, titre of serum pepsinogen, peripheral eosinophilia and eggs per gram (epg). IgA activity in gastric mucus, serum, nasal secretions and saliva were tested against somatic antigen from fourth-stage larvae (L4), somatic antigen from the adult stage and excretory-secretory (E/S) antigen from the adult stage. The results showed a significant correlation between serum IgA and gastric mucus (P<0.01) as well as in nasal secretions (P<0.01). We found negative correlations between IgA activity in gastric mucus with the eggs in utero and with adult female length. Furthermore there were also strong relationships between the peripheral eosinophilia with serum (P<0.01) and gastric mucus IgA activity (P<0.01). Moreover serum pepsinogen and the number of L4 at slaughter were related (P<0.01). PMID- 16048641 TI - Effect of a low protein diet on the resistance of Churra sheep to Teladorsagia circumcincta. AB - SUMMARY The aim of this study was to understand the influence of a low protein diet on the resistance of the Churra breed sheep to infection with Teladorsagia circumcincta. A previous experiment in our department demonstrated significant differences between resistant and susceptible sheep infected with T. circumcincta, in eggs per gram of faeces, serum pepsinogen, serum IgA and peripheral eosinophilia (unpublished). The sheep were fed with a balanced protein diet. In the current assay there were significant differences between the resistant and susceptible groups in egg output, serum pepsinogen and worm burden. The optical densities (OD) of the IgA in blood samples, nasal secretions and saliva were tested throughout the study and the differences between groups were not significant. At necropsy adult worm length, the number of eggs in utero in adult females, the number of fourth stage larvae and the OD of gastric mucus IgA did not differ between groups either. Moreover, due to diet there was a loss of weight in both groups but this was not significant. Our results support the view that low protein diets could influence immune function and as a result resistant genotypes do not show any superiority in comparison with susceptible ones. PMID- 16048642 TI - Host humoral immune response to Leishmania lipid-binding protein. AB - SUMMARY We report on the use of Leishmania donovani lipid-binding proteins (LBPs) as antigens capable of being recognized by serum from immunocompetent patients from southern Spain suffering from visceral leishmaniasis and from Peruvian patients with localized cutaneous leishmaniasis caused by Leishmania braziliensis. The absorbance found by immunoenzymatic techniques gave significantly different results for the serum samples from patients with and without leishmaniasis. Specificity by ELISA testing was 93.2% and sensibility 100%. Dot blots from human patient serum samples or naturally infected dogs from Spain gave similarly significant results. All the human serum samples from individuals with visceral leishmaniasis and the Leishmania-positive canine samples recognized two bands, with molecular weights of 8 and 57 kDa. The serum from individuals with cutaneous leishmaniasis caused by L. braziliensis recognized an additional band of 16 kDa. We discuss the role of Leishmania FABP and compare the immunological reactions found with serum samples from other protozoan infections such as toxoplasma and Chagas as well as bacterial infections such as tuberculosis and syphilis. PMID- 16048643 TI - Antigens from Ascaris suum trigger in vitro macrophage NO production. AB - SUMMARY We investigated the in vitro effect of total excretory/secretory and somatic antigens from Ascaris suum adults (ESA and SA) and larvae 3 (ESL3 and SL3), and of 10 purified protein fractions from ESA components on rat alveolar macrophage nitric oxide (NO) production. Our results showed that in vitro incubation of macrophages with SA and SL3 antigens of A. suum did not result in NO release from cells, whereas incubation with ESA or ESL3 antigens resulted in the stimulation of NO production by these cells, both in a specific (inhibited by L-NAME and L-canavanine) and dose-dependent manner. In addition, we could demonstrate that a purified ESA fraction consisting of three Coomassie-stained bands of approximately 37, 44 and 46 kDa is involved in the in vitro triggering of NO production by host cells. These three bands were subjected to MALDI-peptide mass fingerprint, showing similarities with phosphoglycerate kinase, elongation factor Tu and enolase molecules, respectively. Future studies will focus on the characterization of these parasite-derived molecules. PMID- 16048644 TI - Storing, linking, and mining microarray databases using SRS. AB - BACKGROUND: SRS (Sequence Retrieval System) has proven to be a valuable platform for storing, linking, and querying biological databases. Due to the availability of a broad range of different scientific databases in SRS, it has become a useful platform to incorporate and mine microarray data to facilitate the analyses of biological questions and non-hypothesis driven quests. Here we report various solutions and tools for integrating and mining annotated expression data in SRS. RESULTS: We devised an Auto-Upload Tool by which microarray data can be automatically imported into SRS. The dataset can be linked to other databases and user access can be set. The linkage comprehensiveness of microarray platforms to other platforms and biological databases was examined in a network of scientific databases. The stored microarray data can also be made accessible to external programs for further processing. For example, we built an interface to a program called Venn Mapper, which collects its microarray data from SRS, processes the data by creating Venn diagrams, and saves the data for interpretation. CONCLUSION: SRS is a useful database system to store, link and query various scientific datasets, including microarray data. The user-friendly Auto-Upload Tool makes SRS accessible to biologists for linking and mining user-owned databases. PMID- 16048645 TI - Patients' views on outcome following head injury: a qualitative study. AB - BACKGROUND: Head injuries are a common occurrence, with continuing care in the years following injury being provided by primary care teams and a variety of speciality services. The literature on outcome currently reflects areas considered important by health-care professionals, though these may differ in some respects from the views of head injured individuals themselves. Our study aimed to identify aspects of outcome considered important by survivors of traumatic head injury. METHODS: Thirty-two individuals were interviewed, each of whom had suffered head injury between one and ten years previously from which they still had residual difficulties. Purposive sampling was used in order to ensure that views were represented from individuals of differing age, gender and level of disability. These interviews were fully transcribed and analysed qualitatively by a psychologist, a sociologist and a psychiatrist with regular meetings to discuss the coding. RESULTS: Aspects of outcome mentioned by head injury survivors which have received less attention previously included: specific difficulties with group conversations; changes in physical appearance due to scarring or weight change; a sense of loss for the life and sense of self that they had before the injury; and negative reactions of others, often due to lack of understanding of the consequences of injury amongst both family and general public. CONCLUSION: Some aspects of outcome viewed as important by survivors of head injury may be overlooked by health professionals. Consideration of these areas of outcome and the development of suitable interventions should help to improve functional outcome for patients. PMID- 16048646 TI - Calcitonin-producing well-differentiated neuroendocrine carcinoma (carcinoid tumor) of the urinary bladder: case report. AB - BACKGROUND: The occurrence of calcitonin-secreting primary carcinoid tumor of the urinary bladder is extremely rare. CASE PRESENTATION: The case of a 68-year-old male with carcinoid tumor arising in the urinary bladder is presented. Transurethral resection of a polypoid small tumor 0.4 cm in diameter was performed. Immunohistochemical study using neuroendocrine markers allowed a straightforward diagnosis of a low-grade neuroendocrine carcinoma (carcinoid tumor) of the urinary bladder. Immunohistochemistry demonstrated calcitonin immunoreactivity in the most of the tumor cells. CONCLUSION: This tumor shows specific clinical, macroscopical and histological features and must be considered in the differential diagnosis of bladder neoplasms. PMID- 16048647 TI - Role of contractile prostaglandins and Rho-kinase in growth factor-induced airway smooth muscle contraction. AB - BACKGROUND: In addition to their proliferative and differentiating effects, several growth factors are capable of inducing a sustained airway smooth muscle (ASM) contraction. These contractile effects were previously found to be dependent on Rho-kinase and have also been associated with the production of eicosanoids. However, the precise mechanisms underlying growth factor-induced contraction are still unknown. In this study we investigated the role of contractile prostaglandins and Rho-kinase in growth factor-induced ASM contraction. METHODS: Growth factor-induced contractions of guinea pig open-ring tracheal preparations were studied by isometric tension measurements. The contribution of Rho-kinase, mitogen-activated protein kinase (MAPK) and cyclooxygenase (COX) to these reponses was established, using the inhibitors Y 27632 (1 microM), U-0126 (3 microM) and indomethacin (3 microM), respectively. The Rho-kinase dependency of contractions induced by exogenously applied prostaglandin F2alpha (PGF2alpha) and prostaglandin E2 (PGE2) was also studied. In addition, the effects of the selective FP-receptor antagonist AL-8810 (10 microM) and the selective EP1-antagonist AH-6809 (10 microM) on growth factor induced contractions were investigated, both in intact and epithelium-denuded preparations. Growth factor-induced PGF2alpha-and PGE2-release in the absence and presence of Y-27632, U-0126 and indomethacin, was assessed by an ELISA-assay. RESULTS: Epidermal growth factor (EGF)-and platelet-derived growth factor (PDGF) induced contractions of guinea pig tracheal smooth muscle preparations were dependent on Rho-kinase, MAPK and COX. Interestingly, growth factor-induced PGF2alpha-and PGE2-release from tracheal rings was significantly reduced by U 0126 and indomethacin, but not by Y-27632. Also, PGF2alpha-and PGE2-induced ASM contractions were largely dependent on Rho-kinase, in contrast to other contractile agonists like histamine. The FP-receptor antagonist AL-8810 (10 microM) significantly reduced (approximately 50 %) and the EP1-antagonist AH-6809 (10 microM) abrogated growth factor-induced contractions, similarly in intact and epithelium-denuded preparations. CONCLUSION: The results indicate that growth factors induce ASM contraction through contractile prostaglandins - not derived from the epithelium - which in turn rely on Rho-kinase for their contractile effects. PMID- 16048648 TI - Planar Tc99m--sestamibi scintimammography should be considered cautiously in the axillary evaluation of breast cancer protocols: results of an international multicenter trial. AB - BACKGROUND: Lymph node status is the most important prognostic indicator in breast cancer in recently diagnosed primary lesion. As a part of an interregional protocol using scintimammography with Tc99m compounds, the value of planar Tc99m sestamibi scanning for axillary lymph node evaluation is presented. Since there is a wide range of reported values, a standardized protocol of planar imaging was performed. METHODS: One hundred and forty-nine female patients were included prospectively from different regions. Their mean age was 55.1 +/- 11.9 years. Histological report was obtained from 2.987 excised lymph nodes from 150 axillas. An early planar chest image was obtained at 10 min in all patients and a delayed one in 95 patients, all images performed with 740-925 MBq dose of Tc99m sestamibi. Blind lecture of all axillary regions was interpreted by 2 independent observers considering any well defined focal area of increased uptake as an involved axilla. Diagnostic values, 95% confidence intervals [CI] and also likelihood ratios (LR) were calculated. RESULTS: Node histology demonstrated tumor involvement in 546 out of 2987 lymph nodes. Sestamibi was positive in 30 axillas (25 true-positive) and negative in 120 (only 55 true-negative). The sensitivity corresponded to 27.8% [CI = 18.9-38.2] and specificity to 91.7% [81.6 97.2]. The positive and negative LR were 3.33 and 0.79, respectively. There was no difference between early and delayed images. Sensitivity was higher in patients with palpable lesions. CONCLUSION: This work confirmed that non tomographic Tc99m sestamibi scintimammography had a very low detection rate for axillary lymph node involvement and it should not be applied for clinical assessment of breast cancer. PMID- 16048649 TI - Dephosphorylation of CDK9 by protein phosphatase 2A and protein phosphatase-1 in Tat-activated HIV-1 transcription. AB - BACKGROUND: HIV-1 Tat protein recruits human positive transcription elongation factor P-TEFb, consisting of CDK9 and cyclin T1, to HIV-1 transactivation response (TAR) RNA. CDK9 is maintained in dephosphorylated state by TFIIH and undergo phosphorylation upon the dissociation of TFIIH. Thus, dephosphorylation of CDK9 prior to its association with HIV-1 preinitiation complex might be important for HIV-1 transcription. Others and we previously showed that protein phosphatase-2A and protein phosphatase-1 regulates HIV-1 transcription. In the present study we analyze relative contribution of PP2A and PP1 to dephosphorylation of CDK9 and to HIV-1 transcription in vitro and in vivo. RESULTS: In vitro, PP2A but not PP1 dephosphorylated autophosphorylated CDK9 and reduced complex formation between P-TEFb, Tat and TAR RNA. Inhibition of PP2A by okadaic acid inhibited basal as well as Tat-induced HIV-1 transcription whereas inhibition of PP1 by recombinant nuclear inhibitor of PP1 (NIPP1) inhibited only Tat-induced transcription in vitro. In cultured cells, low concentration of okadaic acid, inhibitory for PP2A, only mildly inhibited Tat-induced HIV-1 transcription. In contrast Tat-mediated HIV-1 transcription was strongly inhibited by expression of NIPP1. Okadaic acid induced phosphorylation of endogenous as well transiently expressed CDK9, but this induction was not seen in the cells expressing NIPP1. Also the okadaic acid did not induce phosphorylation of CDK9 with mutation of Thr 186 or with mutations in Ser-329, Thr-330, Thr-333, Ser-334, Ser-347, Thr-350, Ser-353, and Thr-354 residues involved in autophosphorylation of CDK9. CONCLUSION: Our results indicate that although PP2A dephosphorylates autophosphorylated CDK9 in vitro, in cultured cells PP1 is likely to dephosphorylate CDK9 and contribute to the regulation of activated HIV 1 transcription. PMID- 16048650 TI - Women's quality of life is decreased by acute cystitis and antibiotic adverse effects associated with treatment. AB - BACKGROUND: Although acute cystitis is a common infection in women, the impact of this infection and its treatment on women's quality of life (QOL) has not been previously described. OBJECTIVES: To evaluate QOL in women treated for acute cystitis, and describe the relationship between QOL, clinical outcome and adverse events of each of the interventions used in the study. METHODS: DESIGN: Randomized, open-label, multicenter, treatment study. SETTING: Two family medicine outpatient clinics in Iowa. PATIENTS: One-hundred-fifty-seven women with clinical signs and symptoms of acute uncomplicated cystitis. INTERVENTION: Fifty two patients received trimethoprim/sulfamethoxazole 1 double-strength tablet twice daily for 3 days, 54 patients received ciprofloxacin 250 mg twice daily for 3 days and 51 patients received nitrofurantoin 100 mg twice daily for 7 days. MEASUREMENTS: QOL was assessed at the time of enrollment and at 3, 7, 14 and 28 days after the initial visit. QOL was measured using a modified Quality of Well Being scale, a validated, multi-attribute health scale. Clinical outcome was assessed by telephone interview on days 3, 7, 14 and 28 using a standardized questionnaire to assess resolution of symptoms, compliance with the prescribed regimen, and occurrence of adverse events. RESULTS: Patients experiencing a clinical cure had significantly better QOL at days 3 (p = 0.03), 7 (p < 0.001), and 14 (p = 0.02) compared to patients who failed treatment. While there was no difference in QOL by treatment assignment, patients experiencing an adverse event had lower QOL throughout the study period. Patients treated with ciprofloxacin appeared to experience adverse events at a higher rate (62%) compared to those treated with TMP/SMX (45%) and nitrofurantoin (49%), however the difference was not statistically significant (p = 0.2). CONCLUSION: Patients experiencing cystitis have an increase in their QOL with treatment. Those experiencing clinical cure have greater improvement in QOL compared to patients fail therapy. While QOL is improved by treatment, those reporting adverse events have lower overall QOL compared to those who do not experience adverse events. This study is important in that it suggests that both cystitis and antibiotic treatment can affect QOL in a measurable way. PMID- 16048651 TI - Artificial neural networks allow the use of simultaneous measurements of Alzheimer disease markers for early detection of the disease. AB - BACKGROUND: Previous studies have shown that in platelets of mild Alzheimer Disease (AD) patients there are alterations of specific APP forms, paralleled by alteration in expression level of both ADAM 10 and BACE when compared to control subjects. Due to the poor linear relation among each key-element of beta-amyloid cascade and the target diagnosis, the use of systems able to afford non linear tasks, like artificial neural networks (ANNs), should allow a better discriminating capacity in comparison with classical statistics. OBJECTIVE: To evaluate the accuracy of ANNs in AD diagnosis. METHODS: 37 mild-AD patients and 25 control subjects were enrolled, and APP, ADM10 and BACE measures were performed. Fifteen different models of feed-forward and complex-recurrent ANNs (provided by Semeion Research Centre), based on different learning laws (back propagation, sine-net, bi-modal) were compared with the linear discriminant analysis (LDA). RESULTS: The best ANN model correctly identified mild AD patients in the 94% of cases and the control subjects in the 92%. The corresponding diagnostic performance obtained with LDA was 90% and 73%. CONCLUSION: This preliminary study suggests that the processing of biochemical tests related to beta-amyloid cascade with ANNs allows a very good discrimination of AD in early stages, higher than that obtainable with classical statistics methods. PMID- 16048652 TI - Heart rate variability in non-apneic snorers and controls before and after continuous positive airway pressure. AB - BACKGROUND: We hypothesized that sympathetic nervous system activity (SNSA) is increased and parasympathetic nervous system activity (PNSA) is decreased during non-rapid eye movement (NREM) sleep in non-apneic, otherwise healthy, snoring individuals compared to control. Moreover, we hypothesized that these alterations in snoring individuals would be more evident during non-snoring than snoring when compared to control. METHODS: To test these hypotheses, heart rate variability was used to measure PNSA and SNSA in 11 normotensive non-apneic snorers and 12 control subjects before and 7-days after adapting to nasal continuous positive airway pressure (nCPAP). RESULTS: Our results showed that SNSA was increased and PNSA was decreased in non-apneic snorers during NREM compared to control. However, these changes were only evident during the study in which snoring was eliminated with nCPAP. Conversely, during periods of snoring SNSA and PNSA were similar to measures obtained from the control group. Additionally, within the control group, SNSA and PNSA did not vary before and after nCPAP application. CONCLUSION: Our findings suggest that long-lasting alterations in autonomic function may exist in snoring subjects that are otherwise healthy. Moreover, we speculate that because of competing inputs (i.e. inhibitory versus excitatory inputs) to the autonomic nervous system during snoring, the full impact of snoring on autonomic function is most evident during non-snoring periods. PMID- 16048653 TI - The GuideLine Implementability Appraisal (GLIA): development of an instrument to identify obstacles to guideline implementation. AB - BACKGROUND: Clinical practice guidelines are not uniformly successful in influencing clinicians' behaviour toward best practices. Implementability refers to a set of characteristics that predict ease of (and obstacles to) guideline implementation. Our objective is to develop and validate a tool for appraisal of implementability of clinical guidelines. METHODS: Indicators of implementability were identified from the literature and used to create items and dimensions of the GuideLine Implementability Appraisal (GLIA). GLIA consists of 31 items, arranged into 10 dimensions. Questions from 9 of the 10 dimensions are applied individually to each recommendation of the guideline. Decidability and Executability are critical dimensions. Other dimensions are Global, Presentation and Formatting, Measurable Outcomes, Apparent Validity, Flexibility, Effect on Process of Care, Novelty/Innovation, and Computability. We conducted a series of validation activities, including validation of the construct of implementability, expert review of content for clarity, relevance, and comprehensiveness, and assessment of construct validity of the instrument. Finally, GLIA was applied to a draft guideline under development by national professional societies. RESULTS: Evidence of content validity and preliminary support for construct validity were obtained. The GLIA proved to be useful in identifying barriers to implementation in the draft guideline and the guideline was revised accordingly. CONCLUSION: GLIA may be useful to guideline developers who can apply the results to remedy defects in their guidelines. Likewise, guideline implementers may use GLIA to select implementable recommendations and to devise implementation strategies that address identified barriers. By aiding the design and operationalization of highly implementable guidelines, our goal is that application of GLIA may help to improve health outcomes, but further evaluation will be required to support this potential benefit. PMID- 16048654 TI - Off-trial evaluation of bisphosphonates in patients with metastatic breast cancer. AB - BACKGROUND: Bisphosphonate therapy has been readily accepted as standard of care for individuals with bone metastases from breast cancer. In this study we determined whether the proportion of patients experiencing a skeletal related event (SRE) in a clinical practice population was similar to that observed in phase III randomized controlled studies. METHODS: A retrospective chart review was conducted of 110 patients receiving intravenous bisphosphonates for advanced breast cancer. The proportion of patients experiencing at least one SRE after 12 months of therapy was determined. SRE included vertebral or non-vertebral fracture, cord compression, surgery and/or radiotherapy to bone. RESULTS: The proportion of patients who had an SRE was 30% (28 individuals) and the median time to first event was greater than 350 days. Non-vertebral events and radiotherapy were the most frequent type of SRE, while cord compression and hypercalcaemia were rare (1%). Most patients in the study had bone-only disease (58.2%) and most had multiple bone lesions. In the first 12 months the mean duration of exposure to intravenous bisphosphonates was 261 days and most patients remained on treatment until just before death (median 27 days). CONCLUSION: This study suggests that the rate of clinically relevant SREs is substantially lower than the event rate observed in phase III clinical trials. We attribute this lower rate to observational bias. In the clinical trial setting it is possible that over-detection of skeletal events occurs due to the utilisation of regular skeletal survey or radionucleotide bone scan, whereas these procedures are not routine in clinical practice. Phase IV observational studies need to be conducted to determine the true benefits of bisphosphonate therapy in order to implement rationale use of bisphosphonates. PMID- 16048655 TI - Palmoplantar keratoderma is associated with esophagus squamous cell cancer in Van region of Turkey: a case control study. AB - BACKGROUND: Esophagus squamous cell cancer (ESCC) is the most common cancer in women with 20.2% and second in men with 10.7% relative frequency among all cancer cases diagnosed in Van Region in the east of Turkey. Ninety percent of all esophageal cancer cases are ESCC and 20-30% of them have family history of esophageal cancer. The most clear defined hereditary predisposition associated with ESCC is palmoplantar keratoderma (PPK). To examine the relationship between ESCC and PPK, we have carried out this case control study. METHODS: The case group consisted of 48 subjects who had new diagnosis of ESCC and did not receive any chemo or radiotherapy. The control group consisted of 96 healthy individuals who were visitors of their relatives in the hospital. Two control persons who matched for age, gender, living place (urban/rural) and region were selected for each case. All subjects were evaluated for PPK by dermatologist. Evaluation was graded as none, mild, evident and severe. None and mild subjects were classified as negative for PPK; and others as positive. Relationship between ESCC and PPK was evaluated with odds ratios and confidence intervals for cases with or without family history of ESCC. RESULTS: The PPK frequencies were 92.3% in ESCC cases with family history, 62.5% in ESCC cases without family history, 70.8% in all ESCC cases, and 28.1% in the control group. Odds ratios for cases with or without family history of esophageal cancer, and for the whole case group were found as 30.7 (95%CI = 3.8-247.4), 4.3 (95%CI = 1.9-9.8) and 6.2 (95%CI = 2.9-13.3) respectively. CONCLUSION: Presence of PPK lesions represents genetic susceptibility for ESCC. This susceptibility for ESCC is the highest among those who have PPK lesions and a positive family history of esophageal cancer. Furthermore, a PPK sufferer has an increased risk of developing ESCC even if there is no family history of esophageal cancer. PMID- 16048656 TI - Intensity modulated radiotherapy for high risk prostate cancer based on sentinel node SPECT imaging for target volume definition. AB - BACKGROUND: The RTOG 94-13 trial has provided evidence that patients with high risk prostate cancer benefit from an additional radiotherapy to the pelvic nodes combined with concomitant hormonal ablation. Since lymphatic drainage of the prostate is highly variable, the optimal target volume definition for the pelvic lymph nodes is problematic. To overcome this limitation, we tested the feasibility of an intensity modulated radiation therapy (IMRT) protocol, taking under consideration the individual pelvic sentinel node drainage pattern by SPECT functional imaging. METHODS: Patients with high risk prostate cancer were included. Sentinel nodes (SN) were localised 1.5-3 hours after injection of 250 MBq 99mTc-Nanocoll using a double-headed gamma camera with an integrated X-Ray device. All sentinel node localisations were included into the pelvic clinical target volume (CTV). Dose prescriptions were 50.4 Gy (5 x 1.8 Gy / week) to the pelvis and 70.0 Gy (5 x 2.0 Gy / week) to the prostate including the base of seminal vesicles or whole seminal vesicles. Patients were treated with IMRT. Furthermore a theoretical comparison between IMRT and a three-dimensional conformal technique was performed. RESULTS: Since 08/2003 6 patients were treated with this protocol. All patients had detectable sentinel lymph nodes (total 29). 4 of 6 patients showed sentinel node localisations (total 10), that would not have been treated adequately with CT-based planning ('geographical miss') only. The most common localisation for a probable geographical miss was the perirectal area. The comparison between dose-volume-histograms of IMRT- and conventional CT planning demonstrated clear superiority of IMRT when all sentinel lymph nodes were included. IMRT allowed a significantly better sparing of normal tissue and reduced volumes of small bowel, large bowel and rectum irradiated with critical doses. No gastrointestinal or genitourinary acute toxicity Grade 3 or 4 (RTOG) occurred. CONCLUSION: IMRT based on sentinel lymph node identification is feasible and reduces the probability of a geographical miss. Furthermore, IMRT allows a pronounced sparing of normal tissue irradiation. Thus, the chosen approach will help to increase the curative potential of radiotherapy in high risk prostate cancer patients. PMID- 16048657 TI - Hospital mealtimes: action research for change? AB - Poor nutritional care within the hospital setting continues despite decades of work chronicling and measuring the problems. To address the problem changes in practice have been attempted to improve the patients' experience of mealtimes. In order to implement patient-centred mealtimes for older patients by changing the focus from institutional convenience to one that focuses on the requirements of the patients, an action research approach has been used that focuses on action and change, and thus appears to have much to offer those who seek to change practice. The present paper focuses on the first two phases in a three-phase approach. In phase one the nature of everyday mealtime care and the wider context are explored using focus groups, interviews and observations. The data fall into three main themes that all impact on patients' experiences of mealtimes: institutional and organisational constraints; mealtime care and nursing priorities; eating environment. Following feedback of phase 1 findings to staff and identification of areas of concern a model of practice development was selected to guide the change process of the second phase. Changes to mealtime nursing practice and the ward environment have been made, indicating that action research has the potential to improve the mealtime care of patients. PMID- 16048658 TI - Death, morbidity and economics are the only end points for trials. AB - In order to determine whether surrogate markers predict clinical outcome, randomized controlled trials (RCT) of nutrition support v. no nutrition support that have reported at least one clinical outcome (mortality, infections, total complications, or duration of hospitalization) and at least one nutritional outcome (energy or protein intake, weight gain, N balance, albumin, prealbumin, transferrin, three anthropometric measures, skin testing, lymphocyte count) were assessed for concordance. If changes in nutritional markers predict clinical outcome, changes in both outcomes should go in the same direction. Concordance is defined as both outcomes changing in the same direction or both outcomes showing no difference. Discordance is defined as one outcome changing and the other not (partial) or both outcomes changing in opposite directions (complete). Ninety nine RCT were identified, of which most were underpowered to see statistically significant changes, especially in clinical outcomes. Thus, the results were analysed only in relation to the direction of the respective changes in outcomes. Forty-eight comparisons (4 x 12) were made. The rates of concordance were < or =50% in forty-one of forty-eight comparisons; the rate was never >75%. A complete discordance rate of > or =25% was present in forty-three (> or =50% in thirteen) of the forty-eight comparisons. The discordance was usually a result of the nutritional outcome being better than the clinical outcome. Changes in nutritional markers do not predict clinical outcomes. Before adopting any surrogate marker as an end point for a clinical trial, it has to be known that improving it will result in patient benefit. PMID- 16048659 TI - Physiological functions should be considered as true end points of nutritional intervention studies. AB - With the beginning of this millennium it has become fashionable to only follow 'evidence-based' practices. This generally-accepted approach cruelly negates experience or intelligent interpretation of pathophysiology. Another problem is that the great 'meta-analysts' of the present era only accept end points that they consider 'hard'. In the metabolic and nutritional field these end points are infection-related morbidity and mortality, and all other end points are considered 'surrogate'. The aim of this presentation is to prove that this claim greatly negates the contribution of more-fundamentally-oriented research, the fact that mortality has multifactorial causes, and that infection is a crude measure of immune function. The following problems should be considered: many populations undergoing intervention have low mortality, requiring studies with thousands of patients to demonstrate effects of intervention on mortality; nutrition is only in rare cases primary treatment, and in many populations is a prerequisite for survival rather than a therapeutic modality; once the effect of nutritional support is achieved, the extra benefit of modulation of the nutritional support regimen can only be modest; cost-benefit is not a valid end point, because the better it is done the more it will cost; morbidity and mortality are crude end points for the effect of nutritional intervention, and are influenced by many factors. In fact, it is a yes or no factor. In the literature the most important contributions include new insights into the pathogenesis of disease, the diminution of disease-related adverse events and/or functional improvement after therapy. In nutrition research the negligence of these end points has precluded the development and validation of functional end points, such as muscle, immune and cognitive functions. Disability, quality of life, morbidity and mortality are directly related to these functional variables. It is, therefore, of paramount importance to validate functional end points and to consider them as primary rather than surrogate end points. PMID- 16048660 TI - Biochemical homeostasis and body growth are reliable end points in clinical nutrition trials. AB - Studies of biochemical homeostasis and/or body growth have been included as outcome variables in most nutrition trials in paediatric patients. Moreover, these outcome variables have provided important insights into the nutrient requirements of infants and children, and continue to do so. Examples of the value of such studies in improving parenteral nutrition, in defining essential fatty acid metabolism and requirements of infants and in defining the protein and energy needs of low-birth-weight infants are discussed. Data from such studies have helped to define the mechanism of metabolic acidosis and hyperammonaemia associated with the use of early crystalline amino acid mixture and, hence, how to prevent these disorders. Such studies have allowed the development of parenteral amino acid mixtures that circumvent grossly abnormal plasma concentrations of most amino acids and appear to be utilized more efficiently. These studies have also helped define micronutrient requirements, including requirements for several such nutrients that had not been previously recognized as essential (e.g. Cr, Se, Mo, alpha-linolenic acid). Studies of body growth have been particularly valuable in defining the nutritional requirements of low-birth weight infants. Finally, studies of metabolic homeostasis coupled with more sophisticated metabolic studies have provided considerable insight into the metabolism of the essential fatty acids, linoleic acid (18:2n-6) and alpha linolenic acid (18:3n-3). Although such studies have not defined the amount of the longer-chain PUFA synthesized from each of these essential fatty acids, i.e. arachidonic acid (20:4n-6) and DHA (22:6n-3), they have shown that the rates of conversion are extremely variable from infant to infant, suggesting a possible explanation of why some studies show developmental advantages from intake of these fatty acids while others do not. PMID- 16048661 TI - Elucidating effective ways to identify and treat malnutrition. AB - There is a clear rationale for elucidating effective ways of identifying and treating disease-related malnutrition (DRM), given the physiological and financial consequences of this common condition and its treatability. Evidence indicates the efficacy of nutritional support methods (oral, tube and intravenous) in increasing total nutritional intake while having little effect on appetite, satiety, appetite mediators (e.g. leptin) and voluntary food intake. When used as the only source of nutrition, artificial nutrition can effectively maintain nutritional intake, and yet many patients find enteral or parenteral feeding alone is unable to relieve distressing appetite sensations, and unusual temporal patterns (including dissociation between hunger and desire to eat) occur. Despite the positive impact of these feeding methods on intake, controversy about whether nutritional support can affect patient outcome has remained. Systematic reviews and meta-analyses indicate that improvements in function and clinical (mortality, complication rates) outcome can occur in a number of patient groups (including hospitalised patients, the elderly, patients who have had gastrointestinal surgery, patients at risk of pressure ulcers). In order to target those patients who will benefit from nutritional support, and overcome the ongoing problem of poor detection and recognition of DRM, simple routine screening to identify risk followed by practical evidence-based treatment is recommended. PMID- 16048662 TI - Use of BMI in the assessment of undernutrition in older subjects: reflecting on practice. AB - In recent years there has been a proliferation of nutrition screening tools but undernutrition remains prevalent amongst older subjects. Screening tools commonly include BMI as the widely-accepted 'gold standard' indicator of malnutrition. Whilst BMI may be an appropriate tool for population studies when it can be measured accurately in research conditions, the use of BMI in clinical practice may mask important weight changes and result in a failure to alert healthcare staff to a nutritional problem. The inclusion of BMI has been identified as a barrier to completing the screening process at ward level. Also, feedback from dietitians working with older subjects indicates that 72.5% of those using BMI express concerns that it is of limited use for practical reasons or that the reference range (20-25 kg/m2) is not appropriate to older subjects. Further evidence questions whether or not BMI is applicable for inclusion in methods used to identify an older subject at risk of undernutrition in a variety of care settings. In view of these findings it is advocated that weight change over a period of time together with clinical judgement is a far superior prognostic indicator of undernutrition. Despite screening, there is evidence that inpatients continue to lose weight before discharge. Further experiential evidence from both community and ward settings suggests that inadequacies in care planning, food provision and a lack of assistance with feeding are common. In order to improve the management of undernutrition in older subjects it is therefore recommended that the focus of attention should be on addressing these practical issues and on the effective monitoring of these processes. PMID- 16048663 TI - Integrated nutrition. AB - There is no branch of medicine in which nutritional considerations do not play some part. Overnutrition, undernutrition or unbalanced nutrition are the major causes of ill health in the world. Conversely, illness causes important nutritional and metabolic problems. The spectrum from lack to excess of nutrients is seamless as a clinical and scientific discipline, the two extremes being linked by the Barker effect by which intrauterine malnutrition and low birth weight predispose to obesity, diabetes and CVD in later life. However, the teaching of nutrition in medical and nursing schools remains sparse. Nutritional care cannot be practised satisfactorily in isolation from other aspects of management, since factors such as drugs, surgery and fluid and electrolyte balance affect nutritional status. Nutritional treatment may also have adverse or beneficial effects according to the composition, amount and mode of delivery of the diet and the clinical context in which it is given. Any benefits of nutritional support may also be negated by shortcomings in other aspects of treatment and must therefore be fully integrated into overall care. One example of this approach is the enhanced recovery after a surgery protocol incorporating immediate pre-operative carbohydrate and early post-operative oral intake with strict attention to zero fluid balance, epidural analgesia and early mobilisation. Other examples include the deleterious effect on surgical outcome of salt and water overload or hyperglycaemia, either of which may negate the benefits of nutritional support. There is a need, therefore, to integrate clinical nutrition more closely, not just into medical and surgical practice, but also into the organisation of health services in the hospital and the community, and into the training of doctors and nurses. Societies originally devoted to parenteral and enteral nutrition need to widen their scope to embrace wider aspects of clinical nutrition. PMID- 16048664 TI - Should food or supplements be used in the community for the treatment of disease related malnutrition? AB - Strategies are needed for community-based treatment of disease-related malnutrition (DRM), which is a common debilitating condition that in the UK is estimated to cost > 7 pounds x 10(9) annually. Whilst dietary fortification and counselling are often used as a first-line treatment for malnutrition, the numbers of dietitians available to undertake and oversee such practices are currently insufficient to address the extent of DRM in primary care. Although dietary fortification and counselling can improve nutritional (primarily energy) intake, the evidence base for this practice is weak and it needs addressing with well-designed trials that assess clinically-relevant outcome measures and costs. Liquid oral nutritional supplements (ONS) are increasingly used in the community, often in combination with dietary counselling. The larger evidence base of trials that have assessed ONS suggests that nutritional intake and some functional outcomes can be improved in some patient groups in the community. Although meta analysis indicates significant reductions in mortality (odds ratio 0.59 (95% CI 0.48, 0.72), n 3258) and complication rates (odds ratio 0.41 (95% CI 0.31, 0.53), n 1710) with ONS v. routine care, few of these studies are community based. Thus, the impact of ONS on clinical outcome, healthcare use and costs requires further assessment. Similarly, the clinical and cost efficacy of other strategies (e.g. sensory enhancement, music, behavioural therapy), alone or in combination with other treatments, requires greater investigation in order to meet the challenge of treating DRM more effectively and cheaply in the future. PMID- 16048665 TI - Practice and problems with gastrostomies. AB - The development of gastrostomy placement has been an important technological advance in the enteral-access field. However, its rapid growth in popularity could be viewed as problematic. The endoscopist or intervention radiologist can no longer act as a technician but requires to follow-up this group of patients in order to determine outcomes that will inform future practice. There has been emphasis on the importance of the multidisciplinary team in informing and assessing patients referred for gastrostomy insertion. Communication between all the professionals caring for a patient and between the carers and the patient allows information to be collated that will determine the benefits and burdens of long-term gastrostomy feeding. At present much of the published experience is limited to the acute care setting. The incidence of complications varies, depending on the investigator's definition of complication and the diagnosis of the patient group. Many reported studies are retrospective, which puts limitations on documentation. Increasing numbers of patients with diverse needs are now being discharged into the community with gastrostomy tubes in place. Whilst the hospital should ensure that written protocols are provided and that all carers involved should have adequate training, it is not unusual for patients to receive mixed messages from the different care teams responsible for their care. In South Glasgow NHS Hospital Division key members of all teams caring for these patients (acute care, community district nurses, learning disabilities team, physical disabilities team and commercial homecare companies) meet regularly to discuss equipment and protocols. The members of this group feel that this approach has improved communication, standardised practice and reduced complications by providing a service that delivers artificial nutrition support but is primarily suited to the patient's disease process. PMID- 16048666 TI - The parent's perspective. AB - The present paper gives an insight from a parent's perspective into the roles of health professionals and service providers in the daily management of a child with complex needs that include enteral feeding. It focuses on the case of a 9 year-old boy and discusses some aspects of his diagnosis and treatment, and the support received. It highlights the need for a multi-agency approach based around the child, in which parents are consulted, the opinions of professionals from the different disciplines are valued by other professionals and professionals do not issue conflicting advice but share knowledge before giving advice. There should be national accessibility to support services and a standardised training programme for carers. Better communication between parents, carers, health professionals and service providers and working together can reduce the stress for the patient and carer, and put less strain on much-needed resources. PMID- 16048667 TI - LIPGENE: an integrated approach to tackling the metabolic syndrome. AB - The prevalence of obesity, overweight and type 2 diabetes are increasing in all regions of Europe. Obesity is already commonplace, affecting 10-20% of men and 10 25% of women, and by 2010 approximately 31 million of the population will require treatment for diabetes and its related complications, including the 'metabolic syndrome' (a cluster of cardiovascular risk factors). Associated health and social welfare costs are expected to rise to economically unsustainable levels in Europe as a direct result of these trends. Effective strategies are needed to tackle this major public health problem and to decrease dependence on medical management. These issues are the focus of LIPGENE (FOOD-CT-2003-505944), funded by the European Commission, which will investigate the interactions between dietary constituents and the genome in the development of chronic diseases, such as the metabolic syndrome, and will utilise new technologies to identify novel solutions. The LIPGENE consortium comprises twenty-five research centres across Europe. Features of the 5-year work programme include a major human nutrition intervention study in eight European cities, development of a sustainable vegetable oil product naturally rich in long-chain n-3 fatty acids, and identification of a protocol for feeding dairy cows that will result in milk with a more favourable fatty acid composition. Other work packages will provide a detailed economic analysis of the current and future healthcare costs associated with the metabolic syndrome, and an analysis of consumer attitudes. There is also a dissemination programme associated with the project that features conferences, workshops and associated publications. PMID- 16048668 TI - Metabolic syndrome: what is it and what are the implications? AB - Obesity and overweight are linked with a cluster of metabolic and vascular disorders that have been termed the metabolic syndrome. Although there is not yet a universally-accepted set of diagnostic criteria, most expert groups agree that the syndrome is characterised by impaired insulin sensitivity and hyperglycaemia, dyslipidaemia (elevated blood triacyglycerols with depressed HDL-cholesterol), abdominal obesity and hypertension. Based on existing published criteria estimates suggest that the syndrome affects a substantial percentage of the middle-aged and elderly populations of most European countries (10-20%) and confers increased risk of type 2 diabetes (2-8.8-fold) and CVD (1.5-6-fold), as well as having a marked effect on morbidity. Although the pathophysiology is incompletely understood, insulin resistance and abdominal obesity are central to subsequent abnormalities in circulating glucose and lipoproteins, and vascular function that lead to type 2 diabetes, atherosclerosis and CVD. The link between metabolic syndrome, type 2 diabetes and CVD, as well as inability to reverse the present rising rates of obesity, will lead to economically-unsustainable costs of health care in the next 10-20 years. Preventative strategies for metabolic syndrome are required to slow rates of progression and to reduce dependence on costly medical management. A notable development is recent evidence that shows that diet and exercise are more effective than drug treatment in preventing the development of type-2 diabetes in high-risk individuals. The LIPGENE project will investigate dietary fat quality as a strategy for the prevention of metabolic syndrome and identify food chain approaches that can support consumer attempts to alter their dietary patterns. PMID- 16048669 TI - The prevalence and costs of obesity in the EU. AB - The present paper reports on results obtained as part of the economics module of the LIPGENE project. It reviews recent trends in obesity in the fifteen member states of the EU (the member states before enlargement in 2004) and concludes that in 2002 for both adult men and women at least half the fifteen member states of the EU had obesity levels of >20%. In the same year the total direct and indirect annual costs of obesity in the fifteen member states of the EU were 32,800 euro x 10(6). The provision of 'healthy' foods with an adapted lipid profile provides one means of mitigating the impact of obesity. However, there are extra costs associated with the supply of such foods, via additional costs of food ingredients, identity preservation costs for premium-value food products and the penalties caused by the inability to exploit economies of scale. The limited scale of the markets for healthy foods is caused by their high selling prices. Also presented is an analysis of the costs of subsidising, throughout the EU, ingredients that improve the fatty acid profile of beef, eggs, poultry meat and low-fat spreads, so that healthier versions of these products sell at the same price as traditional alternatives. The total costs of such subsidies for these four food products would have been 10 euro x 10(9)/year in 2002, which was 30% of the direct and indirect costs of obesity. PMID- 16048670 TI - Psychological underpinnings of metabolic syndrome. AB - Metabolic syndrome (MS) is more common among socio-economically disadvantaged individuals and is associated with certain risky lifestyle practices. MS also appears to be triggered by adverse social circumstances and chronic stress. The present paper reviews accumulating evidence to imply that individuals who have certain personality and behaviour traits are particularly predisposed to develop MS, and brings together theories that relate to possible psychological mechanisms underlying MS. It considers how such factors might interact causally to encourage the development of MS. As part of the EU-funded LIPGENE Integrated Project, multi level modelling will be undertaken to explore potential pathways to MS, taking into consideration the interplay between a range of psycho-social, demographic, cultural and lifestyle factors thought to contribute to the development of MS. Data will be gathered for this purpose from a representative sample of >50-year olds living in Britain (n 1000) and Portugal (n 500). It is anticipated that this information will assist in the development and targetting of future intervention to prevent and treat MS in the normal population. PMID- 16048671 TI - The metabolic syndrome: the crossroads of diet and genetics. AB - The metabolic syndrome is a very common disease associated with an increased risk of type 2 diabetes mellitus (T2DM) and CVD. The clinical characteristics of the metabolic syndrome include insulin resistance, dyslipidaemia, abdominal obesity and hypertension. The diverse clinical characteristics illustrate the complexity of the disease process, which involves several dysregulated metabolic pathways. Thus, multiple genetic targets must be involved in the pathogenesis and progression of the metabolic syndrome. Nevertheless, the human genome has not changed markedly in the last decade but the prevalence of the metabolic syndrome has increased exponentially, which illustrates the importance of gene environmental interactions. There is good evidence that nutrition plays an important role in the development and progression of the metabolic syndrome. Indeed, obesity is a key aetiological factor in the development of the metabolic syndrome. Understanding the biological impact of gene-nutrient interactions will provide a key insight into the pathogenesis and progression of diet-related polygenic disorders, including the metabolic syndrome. The present paper will explore the interactions between genetic background and dietary exposure or nutritional therapy, focusing on the role of dietary fatty acids within the context of nutrient regulation of gene expression and individual responsiveness to dietary therapy. Only with a full understanding of gene-gene, gene-nutrient and gene-nutrient-environment interactions can the molecular basis of the metabolic syndrome be solved to minimise the adverse health effects of obesity and reduce the risk of the metabolic syndrome, and subsequent T2DM and CVD. PMID- 16048672 TI - Modern fat technology: what is the potential for heart health? AB - Saturated and trans-fatty acids raise total cholesterol and LDL-cholesterol and are known to increase the risk of CHD, while dietary unsaturated fatty acids play important roles in maintaining cardiovascular health. Replacing saturated fats with unsaturated fats in the diet often involves many complex dietary changes. Modifying the composition of foods high in saturated fat, particularly those foods that are consumed daily, can help individuals to meet the nutritional targets for reducing the risk of CHD. In the 1960s the Dutch medical community approached Unilever about the technical feasibility of producing margarine with a high-PUFA and low-saturated fatty acid composition. Margarine is an emulsion of water in liquid oil that is stabilised by a network of fat crystals. In-depth expertise of fat crystallisation processes allowed Unilever scientists to use a minimum of solid fat (saturated fatty acids) to structure a maximum level of PUFA rich liquid oil, thus developing the first blood-cholesterol-lowering product, Becel. Over the years the composition of this spread has been modified to reflect new scientific findings and recommendations. The present paper will briefly review the developments in fat technology that have made these improvements possible. Unilever produces spreads that are low in total fat and saturated fat, virtually free of trans-fatty acids and with levels of n-3 and n-6 PUFA that are in line with the latest dietary recommendations for the prevention of CHD. Individuals with the metabolic syndrome have a 2-4-fold increased risk of developing CHD; therefore, these spreads could make a contribution to CHD prevention in this group. In addition, for individuals with the metabolic syndrome the spreads could be further modified to address their unique dyslipidaemia, i.e. elevated blood triacylglycerols and low HDL-cholesterol. Research conducted in the LIPGENE study and other dietary intervention studies will deliver the scientific evidence to justify further modifications in the composition of spreads that are healthy for the heart disease risk factors associated with the metabolic syndrome. PMID- 16048673 TI - The production of very-long-chain PUFA biosynthesis in transgenic plants: towards a sustainable source of fish oils. AB - There is now considerable evidence of the importance of n-3 long-chain PUFA in human health and development. At the same time, the marine fish stocks that serve as the primary sources of these fatty acids are threatened by continued over exploitation. Thus, there is an urgent need to provide a sustainable alternative source of the n-3 long-chain PUFA normally found in fish oils. The possibility of using transgenic plants genetically engineered to synthesise these important fatty acids has recently been demonstrated. The approaches taken to realise this outcome will be discussed, as will their prospects for providing a sustainable resource for the future. PMID- 16048674 TI - The role of animal nutrition in improving the nutritive value of animal-derived foods in relation to chronic disease. AB - Foods derived from animals are an important source of nutrients in the diet; for example, milk and meat together provide about 60 and 55% of the dietary intake of Ca and protein respectively in the UK. However, certain aspects of some animal derived foods, particularly their fat and saturated fatty acid (SFA) contents, have led to concerns that these foods substantially contribute to the risk of CVD, the metabolic syndrome and other chronic diseases. In most parts of Europe dairy products are the greatest single dietary source of SFA. The fatty acid composition of various animal-derived foods is, however, not constant and can, in many cases, be enhanced by animal nutrition. In particular, milk fat with reduced concentrations of the C12-16 SFA and an increased concentration of 18:1 MUFA is achievable, although enrichment with very-long-chain n-3 PUFA is much less efficient. However, there is now evidence that some animal-derived foods (notably milk products) contain compounds that may actively promote long-term health, and research is urgently required to fully characterise the benefits associated with the consumption of these compounds and to understand how the levels in natural foods can be enhanced. It is also vital that the beneficial effects are not inadvertently destroyed in the process of reducing the concentrations of SFA. In the future the role of animal nutrition in creating foods closer to the optimum composition for long-term human health is likely to become increasingly important, but production of such foods on a scale that will substantially affect national diets will require political and financial incentives and great changes in the animal production industry. PMID- 16048675 TI - Biochemistry of plant secondary metabolites and their effects in animals. AB - Plant secondary metabolites, which include a wide variety of phytochemicals, have always been constituents of the diets of man and other animals. Although a high proportion of these phytochemicals have been considered to be of little value in plants (although this view is changing), they have frequently been shown to have adverse effects on animals when ingested. The effects depend to a great extent on the chemistry of the compounds, their concentration in the diet and the amount consumed, and are further dependent on the health status of the animals. Traditionally, most studies of the effects of these compounds on animals have focused on their adverse effects and how to alleviate them. However, recent public concern about the use of synthetic compounds in animal diets to enhance performance and health and welfare issues, coupled with changes in regulations on the use of synthetic medicaments, has stimulated interest and research in the use and effects of phytochemicals in the diets of farmed animals. Phytochemicals vary in their chemistry but can be divided into hydrophilic and hydrophobic compounds, of which a wide variety of polyphenolic and terpenoid compounds, as well as alkaloids, carbohydrates and non-protein amino acids, invoke special interest. The chemistry, biochemistry and mechanisms of action of these compounds in plants and their effects in animals when ingested will be explored. PMID- 16048676 TI - Microsatellites and 16S sequences corroborate phenotypic evidence of trans-Andean variation in the parasitoid Microctonus hyperodae (Hymenoptera: Braconidae). AB - Eight South American geographical populations of the parasitoid Microctonus hyperodae Loan were collected in South America (Argentina, Brazil, Chile and Uruguay) and released in New Zealand for biological control of the weevil Listronotus bonariensis (Kuschel), a pest of pasture grasses and cereals. DNA sequencing (16S, COI, 28S, ITS1, beta-tubulin), RAPD, AFLP, microsatellite, SSCP and RFLP analyses were used to seek markers for discriminating between the South American populations. All of the South American populations were more homogeneous than expected. However, variation in microsatellites and 16S gene sequences corroborated morphological, allozyme and other phenotypic evidence of trans-Andes variation between the populations. The Chilean populations were the most genetically variable, while the variation present on the eastern side of the Andes mountains was a subset of that observed in Chile. PMID- 16048677 TI - The separation of two hymenopteran parasitoids, Tersilochus obscurator and Tersilochus microgaster (Ichneumonidae), of stem-mining pests of winter oilseed rape using DNA, morphometric and ecological data. AB - Tersilochus obscurator Aubert and Tersilochus microgaster (Szepligeti) are larval endoparasitoids of economically-important stem-mining pests of winter oilseed rape (Brassica napus L.) in Europe. They are difficult to separate morphologically. Their hosts are Ceutorhynchus pallidactylus (Marsham) and Psylliodes chrysocephala Linnaeus, respectively. The parasitoids' taxonomic status, identification, host range and phenology were studied using genetic, morphometric and ecological data. The study used 527 female parasitoids from the UK and Germany, either field-collected in emergence traps or reared from field collected host larvae. Two morphometric characters, the ovipositor sheath to first metasomal tergite ratio and the percentage of the mesopleuron spanned by the sternaulus, were measured. A 440 bp section of mitochondrial DNA cytochrome oxidase subunit I (COI) gene was sequenced from 35 parasitoids reared from C. pallidactylus, 20 reared from P. chrysocephala and individuals from two outgroups, Tersilochus heterocerus Thomson and Phradis interstitialis Thomson. Distinct and invariable COI sequences corresponded exclusively to each parasitoid group, confirming that T. obscurator and T. microgaster are discrete species. Measurements of host-reared and COI-sequenced specimens indicated that the ranges of both morphometric characters overlapped between species. Using these ranges as criteria, all but 3.6% of UK specimens and 2% of German specimens were identifiable to species without reference to host or phenology. There were differences in emergence phenology in the UK, adult T. microgaster emerging from winter diapause by 29 March 2000, T. obscurator emerging between 12 April and 24 May 2000. The value of molecular techniques in the identification of closely related parasitoid species is discussed. PMID- 16048678 TI - Nuclear DNA content of the whitefly Bemisia tabaci (Aleyrodidae: Hemiptera) estimated by flow cytometry. AB - The nuclear DNA content of the whitefly Bemisia tabaci (Gennnadius) was estimated using flow cytometry. Male and female nuclei were stained with propidium iodide and their DNA content was estimated using chicken red blood cells and Arabidopsis thaliana L. (Brassicaceae) as external standards. The estimated nuclear DNA content of male and female B. tabaci was 1.04 and 2.06 pg, respectively. These results corroborated previous reports based on chromosome counting, which showed that B. tabaci males are haploid and females are diploid. Conversion between DNA content and genome size (1 pg DNA=980 Mbp) indicate that the haploid genome size of B. tabaci is 1020 Mbp, which is approximately five times the size of the genome of the fruitfly Drosophila melanogaster Meigen. These results provide an important baseline that will facilitate genomics-based research for the B. tabaci complex. PMID- 16048679 TI - Development, survivorship and reproduction of Eretmocerus sp. nr furuhashii (Hymenoptera: Aphelinidae) parasitizing Bemisia tabaci (Hemiptera: Aleyrodidae) on glabrous and non-glabrous host plants. AB - The developmental, survivorship and reproductive biology of Eretmocerus sp. nr furuhashii Rose & Zolnerowich parasitizing Bemisia tabaci (Gennadius) biotype B on tomato, cucumber, eggplant and collard at 26+/-0.5 degrees C was studied. The mean longevity ranged from 6.5 days on tomato to 8.1 days on collard. The mean lifetime fecundities on tomato, cucumber, eggplant and collard were 35.4, 37.3, 41.4 and 46.4 eggs, whereas the mean lifetime fertility was 20.2, 22.7, 28.3 and 33.6 offspring, respectively. Developmental time was longest on cucumber (17.1 days) and shortest on collard (15.9 days). Survival rates varied significantly across host plants. Sex ratio (female symbol:male symbol) varied from 1.71 to 1.93 across the four hosts. The intrinsic rate of increase (rm) was highest (0.157) on collard and lowest on tomato (0.133). Mean generation time (R0) ranged from 14.7 to 20.9 days. All the data demonstrated that the reproductive success and overall performance of E. sp. nr furuhashii increased as the density of leaf hairs declined. As leaf hairs play a key role in determining efficacy, it is unlikely that effective biological control of B. tabaci using E. sp. nr furuhashii will be achieved on non-glabrous crop varieties. PMID- 16048680 TI - Effect of temperature and host plant leaf morphology on the efficacy of two entomopathogenic biocontrol agents of Thrips palmi (Thysanoptera: Thripidae). AB - The efficacy of two entomopathogenic biocontrol agents, Steinernema feltiae (Filipjev) and Verticillium lecanii (Zimmerman) Viegas (reclassified now as Lecanicillium muscarium (Petch) Zare & Gams), against Thrips palmi Karny was investigated. Assessments of the effect of temperature on the efficacy of S. feltiae indicated that higher mortality of T. palmi was recorded at 20 degrees C compared to either 15 or 25 degrees C, whereas significantly higher T. palmi mortality followed application of L. muscarium at 25 degrees C. Testing the control agents efficacy on three host plants; chrysanthemum, sweet pepper and cucumber, under constant temperature and high humidity conditions produced no significant difference in the level of T. palmi larval mortality on each host plant. Incorporating the chemical insecticide imidacloprid with both biological agents in a combined control strategy increased T. palmi juvenile mortality. The potential role of S. feltiae and L. muscarium within integrated pest management programmes for the control of T. palmi is discussed. PMID- 16048682 TI - Effect of host quality of Callosobruchus maculatus (Coleoptera: Bruchidae) on performance of the egg parasitoid Uscana lariophaga (Hymenoptera: Trichogrammatidae). AB - Development and reproductive success of the solitary egg parasitoid Uscana lariophaga Steffan were examined after development in eggs of the bruchid storage pest Callosobruchus maculatus Fabricius reared at either low or high densities on cowpea seeds and laid at day 1 and 4 of maternal life. Both bruchid larval competition and maternal age negatively affected egg size, but the latter more than the former. Uscana lariophaga reared in small hosts developed slower, were smaller and produced fewer eggs compared to parasitoids reared in large hosts. Fecundity of the parasitoid was heavily influenced by host egg size. This was reflected in the values for the intrinsic rate of increase of U. lariophaga, which differed for wasps that developed in host eggs laid by bruchid females of different age. Wasps allocated marginally more female offspring to larger hosts. PMID- 16048681 TI - Anopheles (Cellia) epiroticus (Diptera: Culicidae), a new malaria vector species in the Southeast Asian Sundaicus Complex. AB - Anopheles sundaicus species A of the Southeast Asian A. sundaicus complex is formally named Anopheles epiroticus Linton & Harbach based on DNA sequence differentiation of the whole nuclear ITS2 region and a portion of both the cytochrome b and cytochrome c oxidase I mitochondrial genes. Detailed comparative morphological studies of the adult, larval and pupal stages did not reveal any differential or diagnostic differences that reliably distinguish A. epiroticus from A. sundaicus s.s. Information is provided on the bionomics and systematics of the new species. PMID- 16048683 TI - Restriction fragment length polymorphisms of different DNA regions as genetic markers in the hoverfly Episyrphus balteatus (Diptera: Syrphidae). AB - A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis using mitochondrial (A+T-rich region; mtDNA) and genomic (zen-region; nDNA) DNA was performed on 182 female individuals of Episyrphus balteatus (DeGeer), a widespread aphidophagous hoverfly with supposed migratory behaviour. Specimens originated from 13 sampling sites in six European countries. The analyses revealed 12 and 18 haplotypes, respectively, for the two DNA types, several of them with a wide distribution, although seven and eight haplotypes, respectively, occurred only in one location. In contrast to other studies on mobile insects, the genetic diversity was relatively high. However, lack of population subdivision, low genetic distances between populations, the very high gene flow rates, and the complete lack of isolation by distance suggest that E. balteatus populations are largely connected and that there is an absence of large scale geographic structuring. These results support the hypothesis that E. balteatus is a migratory hoverfly species, capable of moving over large distances. These findings related to the seasonal migrations of this species are discussed. PMID- 16048684 TI - Return migration of Helicoverpa armigera (Lepidoptera: Noctuidae) during autumn in northern China. AB - The autumn migration of Helicoverpa armigera (Hubner) was observed with radar and two types of light-trap at Langfang, Hebei province, China in 2001 and 2002. The sudden increase in the proportion of H. armigera moths in the searchlight trap indicated migration into the area and catches increased 10-fold during the second half of the night due to the landing of migrants before dawn. The moths' migratory flights took place at up to 2000 m above the ground, and moths flew differentially at times, and heights, when favourable (i.e. northerly) winds occurred. This facilitated the maximum displacement of moths towards the south during these 'return' migrations. The moths flew over the radar site at consistently high densities through the night, and the resulting flight durations of c. 10 h, at displacement speeds of 30-33 km h-1, would allow moths emerging in the far northeast of China (i.e. Liaoning and Jilin provinces and the Inner Mongolia autonomous region) to migrate into northern China (Hebei, Shandong and Henan provinces). The association of the seasonal migratory movements of H. armigera with crops in northern China is briefly discussed. PMID- 16048685 TI - Molecular and biochemical characterization of a sand fly population from Sri Lanka: evidence for insecticide resistance due to altered esterases and insensitive acetylcholinesterase. AB - With an increasing incidence of cutaneous leishmaniasis in Sri Lanka, particularly in northern provinces, insecticide-mediated vector control is under consideration. Optimizing such a strategy requires the characterization of sand fly populations in target areas with regard to species composition and extant resistance, among other parameters. Sand flies were collected by human bait and cattle-baited net traps on Delft Island, used as an illegal transit location by many refugees returning to the north of Sri Lanka from southern India where leishmaniasis is endemic. For species identification, genomic DNA was extracted and a fragment of the ribosomal 18S gene amplified. The sequence from all flies analysed matched that of Phlebotomus argentipes Annandale & Brunetti, the primary vector in India and the most likely vector in Sri Lanka. Independent morphological analysis also identified P. argentipes. To establish the current susceptibility status of vector species, data were obtained at the biochemical level, from which potential cross-resistance to alternative insecticides can be predicted. The Delft Island collection was assayed for the activities of four enzyme systems involved in insecticide resistance (acetylcholinesterase, non specific carboxylesterases, glutathione-S-transferases and cytochrome p450 monooxygenases), establishing baselines against which subsequent collections can be evaluated. There was preliminary evidence for elevated esterases and altered acetylcholinesterase in this population, the first report of these resistance mechanisms in sand flies to our knowledge, which probably arose from the malathion-based spraying regimes of the Anti-Malarial Campaign. PMID- 16048686 TI - Gene-flow between populations of cotton bollworm Helicoverpa armigera (Lepidoptera: Noctuidae) is highly variable between years. AB - Both large and small scale migrations of Helicoverpa armigera Hubner in Australia were investigated using AMOVA analysis and genetic assignment tests. Five microsatellite loci were screened across 3142 individuals from 16 localities in eight major cotton and grain growing regions within Australia, over a 38-month period (November 1999 to January 2003). From November 1999 to March 2001 relatively low levels of migration were characterized between growing regions. Substantially higher than average gene-flow rates and limited differentiation between cropping regions characterized the period from April 2001 to March 2002. A reduced migration rate in the year from April 2002 to March 2003 resulted in significant genetic structuring between cropping regions. This differentiation was established within two or three generations. Genetic drift alone is unlikely to drive genetic differentiation over such a small number of generations, unless it is accompanied by extreme bottlenecks and/or selection. Helicoverpa armigera in Australia demonstrated isolation by distance, so immigration into cropping regions is more likely to come from nearby regions than from afar. This effect was most pronounced in years with limited migration. However, there is evidence of long distance dispersal events in periods of high migration (April 2001-March 2002). The implications of highly variable migration patterns for resistance management are considered. PMID- 16048688 TI - Ectopic cervical thymoma in the submandibular region. PMID- 16048689 TI - Atypical diagnostic, endoscopic, and pathologic findings in a case of adenoid cystic carcinoma arising from the sphenoid sinus. PMID- 16048690 TI - Epithelial-myoepithelial carcinoma: a low-grade malignancy in the parotid gland. PMID- 16048691 TI - Mucoepidermoid carcinoma of the trachea in a 4-year-old boy. PMID- 16048692 TI - Superficial temporal arteriovenous fistula confirmed with three-dimensional computed tomography-angiography. PMID- 16048693 TI - Recurrent squamous cell carcinoma of the tongue in pregnancy. PMID- 16048694 TI - Giant cystic parapharyngeal space tumour. PMID- 16048695 TI - Unusual metastatic site in a case of carcinoma of the hypopharynx: nasal tip. PMID- 16048696 TI - Intramuscular metastasis of maxillary squamous cell carcinoma. PMID- 16048697 TI - Benign fibrous histiocytoma of the nose. PMID- 16048698 TI - Cervical prevertebral abscess owing to injection of corticosteroids. PMID- 16048699 TI - Cervical necrotizing fasciitis: an unusual complication of genuine peritonsillar abscess. PMID- 16048700 TI - Nd:YAG laser treatment for hemangioma of the nasal cavity. PMID- 16048701 TI - Collagenous spherulosis in epithelial-myoepithelial carcinoma of the submandibular gland: histologic and immunohistochemical study of a case. PMID- 16048702 TI - Complicated hypopharyngeal perforation in a child owing to internal penetrating trauma by a knitting needle--parents beware. PMID- 16048703 TI - Cerebellopontine angle lipoma presenting with hemifacial spasm: case report and review of the literature. PMID- 16048704 TI - Tuberculosis of buccal mucosa. PMID- 16048705 TI - Giant fibrovascular polyp of the oropharynx: case presentation and literature review. PMID- 16048706 TI - Parathyroid adenoma presenting as paraneoplastic syndrome secondary to renal cell carcinoma. PMID- 16048707 TI - Bilateral diffuse gingival enlargement in a patient with type 1 neurofibromatosis. PMID- 16048708 TI - Sinonasal teratocarcinosarcoma involving the cavernous sinus. PMID- 16048709 TI - Aggressive fibromatosis of the head and neck: case report and review of the literature. PMID- 16048710 TI - Infection in breast implants. PMID- 16048712 TI - HIV and hepatitis G virus/GB virus C co-infection: beneficial or not? PMID- 16048713 TI - Traditional African medicine in the treatment of HIV. PMID- 16048714 TI - Tuskegee revisited. PMID- 16048715 TI - Trees become casualties of war. PMID- 16048717 TI - Antimicrobial resistance in developing countries. Part I: recent trends and current status. AB - The global problem of antimicrobial resistance is particularly pressing in developing countries, where the infectious disease burden is high and cost constraints prevent the widespread application of newer, more expensive agents. Gastrointestinal, respiratory, sexually transmitted, and nosocomial infections are leading causes of disease and death in the developing world, and management of all these conditions has been critically compromised by the appearance and rapid spread of resistance. In this first part of the review, we have summarised the present state of resistance in these infections from the available data. Even though surveillance of resistance in many developing countries is suboptimal, the general picture is one of accelerating rates of resistance spurred by antimicrobial misuse and shortfalls in infection control and public health. Reservoirs for resistance may be present in healthy human and animal populations. Considerable economic and health burdens emanate from bacterial resistance, and research is needed to accurately quantify the problem and propose and evaluate practicable solutions. In part II, to be published next month, we will review potential containment strategies that could address this burgeoning problem. PMID- 16048718 TI - Invasive group A streptococcal disease: should close contacts routinely receive antibiotic prophylaxis? AB - Group A streptococci (Streptococcus pyogenes) causes a wide range of illnesses from non-invasive disease--eg, pharyngitis--to more severe invasive infections- eg, necrotising fasciitis and toxic shock-like syndrome. There remains uncertainty about the risk of secondary cases of invasive disease occurring among close contacts of an index case and how best to manage that risk. We do not consider that currently available evidence justifies the routine administration of chemoprophylaxis to close contacts. We suggest that the appropriate response should be to routinely inform all household contacts of a patient with invasive group A streptococcal disease about the clinical manifestations of invasive disease and to seek immediate medical attention if they develop such symptoms. PMID- 16048719 TI - Rapidly progressive soft tissue infections. AB - Skin and soft tissue infections are among the most common reasons for people to seek medical advice. They also represent one of the most common indications for antimicrobial therapy and account for approximately 7-10% of hospitalisations in North America. Although non-limb and non-life threatening infections may be treated on an out-patient basis with oral antibiotics, patients with more serious acute skin and soft tissue infections may require admission to hospital for management; this decision is especially true if the infection is rapidly progressive. We provide a concise overview of the differential diagnosis and approach to management of community-acquired rapidly progressive skin and soft tissue infections. PMID- 16048720 TI - Mass chemotherapy options to control lymphatic filariasis: a systematic review. AB - Understanding the efficacy of microfilaricidal drugs is important in guiding the global programme for the elimination of lymphatic filariasis as a public-health problem. We did a systematic review of the available literature to determine which currently available drug intervention most effectively decreases circulating Wuchereria bancrofti microfilaria in individuals and populations. 57 randomised studies of drug efficacy were identified. Data were combined and compared using weighted mean effect estimates taking into account the longitudinal nature of the data. Combined treatment with diethylcarbamazine plus ivermectin, diethylcarbamazine plus albendazole, and ivermectin plus albendazole resulted in average microfilarial intensity decreases that were 0.7%, 4.6%, and 12.7% of the pre-treatment values, respectively. Drug combinations containing diethylcarbamazine were the most effective against microfilarial prevalence and intensity relative to single drugs or other combinations. The relative efficacies of drug combinations have not been well documented from existing studies and therefore limit the application of evidenced-based recommendations for chemotherapy-based interventions to control lymphatic filariasis. These results provide valuable estimates of drug effect using existing data, but highlight the need for more comprehensive comparative drug studies. PMID- 16048721 TI - Treatment of chronic hepatitis C infection: one step at a time. AB - Non-A, non-B hepatitis was recognised as an important cause of chronic liver disease long before the aetiological agent-hepatitis C virus-was identified in 1989. Recombinant interferons, initially developed as a treatment for malignancies, proved to be an effective treatment for this disease before the identification of the viral agent. Subsequent testing for hepatitis C virus RNA demonstrated that the virus appeared to be eradicated in a small proportion of treated patients. Treatment regimens have improved dramatically since 1989 with the addition of the oral nucleoside ribavirin and long-acting pegylated interferons to treatment regimens. Currently, more than half of treated patients can achieve durable viral clearance. This clearance is quite a remarkable feat; indeed, eradication is not possible in any other chronic viral infection. Considerable effort continues to be devoted to improving therapeutic regimens to make them more effective and tolerable. Drugs that directly act on the replicative machinery of the virus-protease and polymerase inhibitors-are under development and entering clinical trials in human beings. PMID- 16048722 TI - Extensive abscesses following colonic hydrotherapy. PMID- 16048723 TI - Cutaneous diffuse chromoblastomycosis. PMID- 16048724 TI - Erythropoietin promotes endothelial progenitor cell proliferative and adhesive properties in a PI 3-kinase-dependent manner. AB - OBJECTIVES: Patients with congestive heart failure (CHF) suffer considerable morbidity and mortality despite advances in therapy. Treatment with erythropoietin (Epo) has shown promise in CHF patients, yet its mechanisms of action remain elusive. Endothelial progenitor cells (EPC) contribute to postnatal angiogenesis and vasculogenesis, and Epo was shown to promote EPC mobilization. We explored the effect of chronic treatment with Epo on the numbers and functional properties of EPC in CHF patients. METHODS AND RESULTS: Twenty-eight patients with CHF treated with Epo for a mean period of 28 months were compared to a matched group (n = 28) with regard to the number of circulating hematopoietic and endothelial stem cells (either CD34+, CD34+/CD45+, CD34+/CD133+, CD34+/VEGF-R2+ or CD34+/CD133+/VEGF-R2+) as well as their proliferative and adhesive capacity. In vitro, Epo was added to cultured EPC from healthy subjects to test proliferation and adhesion. No differences were observed in circulating numbers of hematopoietic and endothelial stem cells between CHF patients chronically treated with Epo or untreated. EPC from Epo-treated patients exhibited enhanced proliferation as well as a trend towards adhesion to cultured endothelial cells prior to and following stimulation with TNF-alpha. Addition of Epo to EPC from healthy subjects dose-dependently increased their proliferation and adhesion to fibronectin, cultured endothelial cells, and cardiomyocytes. These effects were significantly reduced in the presence of phosphatidylinositol (PI) 3-kinase inhibitors. CONCLUSIONS: Chronic Epo treatment is associated with an increase in the adhesive and proliferative properties of circulating EPC in patients with CHF. PMID- 16048725 TI - Does plasma nitrite determination by the Griess reaction reflect nitric oxide synthesis? PMID- 16048726 TI - Pretreatment levels of serum squamous cell carcinoma antigen and urine polyamines in women with squamous cell carcinoma of the cervix. AB - OBJECTIVES: To investigate whether pretreatment levels of serum squamous cell carcinoma antigen (SCCA) and urine polyamines can predict lymph node metastases in patients with early stage cervical carcinoma. METHODS: Pretreatment measurement of serum SCCA and urine polyamine levels was carried out for 419 women. Of those women, 104 with stage IB and IIA cancer received radical surgery and had tumor size reassessed postoperatively. RESULTS: The women had increased levels of serum SCCA (>2.0 ng/mL) and elevated urine polyamines (>45 micromol/g of creatinine) with advanced cancer stage (P<0.01). The median SCCA level was significantly higher in women with metastatic disease than that in those without lymph node involvement (3.9 vs. 1.1; P<0.01). Women with nodal involvement also had significantly higher median levels of urine polyamines than those without nodal disease. CONCLUSIONS: Pretreatment measurement of SCCA and urine polyamine levels may help in predicting lymph node metastases in women with early stage cervical carcinoma. PMID- 16048727 TI - Rapid receptor-proximal signaling assays for FcR gamma-containing receptors. AB - Novel, cell-based assays, based on bioluminescence resonance energy transfer, have been developed for FcepsilonRI- and GPVI-FcRgamma complex-mediated signaling at receptor-proximal steps. In a stable transfectant of the HEK-293 cell line expressing human FcepsilonRIalpha, FcepsilonRIbeta, and FcRgamma-GFP2 and Syk(1 265)-Rluc fusion proteins, FcepsilonRI cross-linking markedly increased BRET2 ratios, which are the ratios of GFP2 emission to Rluc emission. These ratios reflect the FcRgamma-GFP2-Syk(1-265)-Rluc interaction in living cells. The signals are specifically inhibited by the Src-family kinase inhibitor PP2. Separately, in transient transfectants expressing GPVI, FcRgamma-GFP2, and Syk(1 265)-Rluc, the GPVI-specific ligand convulxin induced a two-fold increase in the BRET2 ratio and this increase was also inhibited by PP2. Finally, a differential assay was developed which permits the measurement of FcepsilonRI- and GPVI FcRgamma complex-mediated signaling in the same cell. These assays provide useful methods for monitoring FcRgamma-Syk interaction in real time in living cells and may contribute to the understanding of signal regulation through FcRgamma containing receptors. PMID- 16048728 TI - Risk factors for experiencing psychosis during cocaine use: a preliminary report. AB - Cocaine induced psychosis (CIP) is a common, but not universal side effect of cocaine abuse. Factors underlying the development and severity of CIP remain poorly understood. This study tests the hypothesis that earlier age of initiation of regular use may increase the likelihood of developing CIP, or the severity of CIP symptoms. METHODS: Cocaine use history and severity of CIP (if any) were assessed with the Cocaine Experience Questionnaire in 51 abstinent (3 weeks-1 year) cocaine dependent individuals. Subjects were divided into those with high and low CIP severity, and into those with early age of initiation of regular cocaine use, and later age of initiation. Various cutoffs between early and late age of initiation were used, ranging from 15 to 22 years. RESULTS: From ages 17 through 20, controlling for cumulative duration of use, severity scores were significantly higher for the early initiation group than for the later initiation group (p values ranged from 0.031 to 0.036). Cumulative duration of use, but not age of initiation, significantly predicted initial development of CIP (p=0.044). CONCLUSIONS: The data suggest that early age of initiation of regular cocaine use occurring during vulnerable periods of brain development, may lead to increased severity of CIP. PMID- 16048729 TI - Transport of microsporidium Encephalitozoon intestinales spores in sandy porous media. AB - The retention and transport of microsporidium Encephalitozoon intestinales spores in two water-saturated sandy porous media was investigated in this study. The initial breakthrough of the spores in the column effluent occurred essentially simultaneously with that of a non-reactive tracer, indicating no significant velocity enhancement. A large fraction (45-73%) of the spores injected into the columns was not recovered in the effluent, indicating removal from solution through colloid retention processes of attachment and/or straining. The relative significance of attachment and straining to total retention was evaluated in additional experiments. An experiment was conducted with a sieved coarse fraction of porous media for which straining is unlikely to be of significance based on the relative diameters of the spores and porous-medium pores. The spore recovery for this experiment was similar to the recoveries obtained for microsporidia transport in the un-sieved parent porous medium. An additional experiment was conducted with a subsample of the coarse fraction that was acid-washed to reduce potential surface attachment sites. Spore recovery was complete for this experiment. These results suggest surface deposition was the primary removal mechanism in our system. This conclusion is supported by the results of an experiment wherein deionized water was flushed through a column that was previously flushed with electrolyte solution. The effluent spore concentrations were observed to increase upon injection of deionized water, indicating re mobilization of spores upon a change in water chemistry. The measured data were successfully simulated using a mathematical model incorporating colloid filtration. The results of this study suggest that the transport of microspordia in sandy porous media is governed by established colloid-transport processes. PMID- 16048730 TI - The effect of sodium ascorbate on the mechanical properties of hyaluronan-based vascular constructs. AB - Esterified hyaluronic acid (HYAFF) is routinely used for clinical tissue engineering applications such as skin and cartilage. In a previous study we developed a technique for in vitro generation of cylindrical constructs from cellularized HYAFF flat sheets. In the present investigation we studied the possibility to improve mechanical properties of this vascular construct by the addition of sodium ascorbate (SA). Non-woven HYAFF flat sheets were seeded with porcine aortic smooth muscle cells (SMCs) and cultured for 14 or 28 days with standard medium or medium added with SA. In selected experiments HYAFF sheets seeded with SMCs were wrapped to obtain cylindrical shape and then cultured in control medium or SA added medium for up to 28 days. We estimated cell viability for flat sheets, and performed histological examination, analysis of extracellular matrix (ECM) deposition and mechanical tests on tubular constructs. The number of viable cells and ECM deposition increased with time in constructs cultured in the presence of SA, as compared to control group. Moreover, SA improved mechanical properties of the vascular construct lowering material stiffness and increasing tensile strength as compared to untreated controls. The addition of SA to the medium improved cell proliferation and ECM synthesis on this biodegradable material, which leads to the formation of well organized, mechanical resistant tissue-engineered structure. PMID- 16048731 TI - Biocompatibility and remodeling potential of pure arterial elastin and collagen scaffolds. AB - Surgical therapy of cardiovascular disorders frequently requires replacement of diseased tissues with prosthetic devices or grafts. In typical tissue engineering approaches, scaffolds are utilized to serve as templates to support cell growth and remodeling. Decellularized vascular matrices have been previously investigated as scaffolds for tissue engineering. However, cell migration into these scaffolds was inadequate due to the very tight matrix organization specific to the aortic structure. To address this problem, we prepared two types of decellularized scaffolds from porcine vascular tissues. Pure elastin scaffolds and pure collagen scaffolds were prepared by selectively removing the collagen component or elastin, respectively. In the current study, we use a subdermal implantation model to demonstrate that arterial elastin and collagen scaffolds exhibit enhanced potential for repopulation by host cells in vivo. Notably, numerous new collagen fibers and bundles were found within the remodeled elastin scaffolds and new elastin fibers within collagen scaffolds, respectively, clearly indicating their ability to support de novo extracellular matrix synthesis. We also show that biological cues such as growth factors are required for efficient repopulation of elastin and collagen scaffolds. Finally, we bring evidence that these scaffolds can be endothelialized in vitro for thrombosis resistance and thus can serve as promising candidates for cardiovascular tissue engineering. PMID- 16048732 TI - Bronchoalveolar lavage examined by solid phase microextraction, gas chromatography--mass spectrometry and selected ion flow tube mass spectrometry. AB - Samples (210 in total) of broncholaveolar lavages (BALs), obtained from patients hospitalized with pneumonia in various departments of two hospitals, were analysed using the method of solid phase microextraction-gas chromatography (SPME GC) with FID detection. Up to 20% (9% unequivocally, 11% probably) of these samples was found to contain volatile fatty acids (VFAs) in the series from acetic acid to heptanoic acid. Importantly, the presence of these acids indicates the presence of fermenting anaerobic bacteria, which were not detected by the conventional microbiological examination. Other compounds, namely the heptanol and cyclohexanone, were also detected by this method in some samples. Cyclohexanone occurred almost exclusively in samples from patients receiving intensive care with mechanical ventilation, and is suspected to originate from plastic parts of ventilators. Selected representative samples were also analysed using further methods, namely gas chromatography-mass spectrometry (GC-MS) of native and silylated samples, and selected ion flow tube mass spectrometry (SIFT MS). These methods confirmed the identities of above mentioned compounds, and detected numerous other compounds tentatively identified as various alcohols, aldehydes, ketones, esters and hydrogen cyanide, HCN. Most of these compounds occurred in small amounts and their origin and diagnostic significance remains uncertain, except, that is, for the HCN, which indicates the presence of Pseudomonas aeruginosa. PMID- 16048733 TI - Predictive modelling of the individual and combined effect of water activity and temperature on the radial growth of Fusarium verticilliodes and F. proliferatum on corn. AB - The major objective of this study was to develop validated models to describe the effect of a(w) and temperature on the radial growth on corn of the two major fumonisin producing Fusaria, namely Fusarium verticilliodes and F. proliferatum. The growth of these two isolates on corn was therefore studied at water activities between 0.810-0.985 and temperatures between 15 and 30 degrees C. Minimum a(w) for growth was 0.869 and 0.854 for F. verticilliodes and F. proliferatum, respectively. No growth took place at a(w) values equal to 0.831 and 0.838 for F. verticilliodes and F. proliferatum, respectively. The colony growth rates, g (mm d(-1)) were determined by fitting a flexible growth model describing the change in colony diameter (mm) with respect to time (days). Secondary models, relating the colony growth rate with a(w) or a(w) and temperature were developed. A third order polynomial equation and the linear Arrhenius-Davey model were used to describe the combined effect of temperature and a(w) on g. The combined modelling approaches, predicting g (mm d(-1)) at any a(w) and/or temperature were validated on independently collected data. All models proved to be good predictors of the growth rates of both isolates on maize within the experimental conditions. The third order polynomial equation had bias factors of 1.042 and 1.054 and accuracy factors of 1.128 and 1.380 for F. verticilliodes and F. proliferatum, respectively. The linear Arrhenius-Davey model had bias factors of 0.978 and 1.002 and accuracy factors of 1.098 and 1.122 for F. verticilliodes and F. proliferatum, respectively. The results confirm the general finding that a(w) has a greater influence on fungal growth than temperature. The developed models can be applied for the prevention of Fusarium growth on maize and the development of models that incorporate other factors important to mould growth on maize. PMID- 16048734 TI - Effect of zolpidem on sleep architecture and its next-morning residual effect in insomniac patients: a randomized crossover comparative study with brotizolam. AB - This study was conducted to determine the effect of zolpidem (ZOL) 10 mg orally on the sleep architecture and the next-morning residual effect in patients with non-organic insomnia (ICD-10) as compared to the effect of brotizolam (BTM) 0.25 mg orally, a widely used short-acting benzodiazepine (BZD) hypnotic in Japan, in a randomized, crossover comparative study. Fourteen patients with non-organic insomnia (3 males and 11 females; mean age of 54.9+/-S.D. 8.9 years). First three nights with placebo, middle three nights with either ZOL 10 mg or BTM 0.25 mg, and last three nights again with placebo in each session (a total of two sessions). Primary endpoints were polysomnography findings of sleep stages, sleep parameters, and sleep latency (SL) in the morning to examine calculable sleepiness as a residual effect. Secondary endpoint was sleep quality assessed by self-assessment questionnaire. At 150 min after Tmax, both ZOL and BTM significantly increased stage 2 (S2), and ZOL showed significantly longer slow wave sleep (SWS; stage 3+4) as compared to BTM. Stage wake was significantly increased by ZOL at the first withdrawal night and by BTM at the second withdrawal night. ZOL did not affect SL after rising, whereas BTM showed significantly shorter SL. Both drugs reduced the number of nocturnal awakenings and improved subjective sleep quality. The common adverse drug reaction (ADR) was sleepiness (3 patients) in each treatment. All events were mild. No serious adverse events occurred. ZOL is as effective as BTM in improving subjective sleep quality in patients with psychophysiological insomnia (PPI). ZOL has advantages over BTM in having a unique profile of increasing SWS with less next-morning residual effect. PMID- 16048735 TI - Heavy metals extraction from contaminated soil: recovery of the flushing solution. AB - The optimum conditions for the recovery of copper from a contaminated soil by using the soil flushing technique are evaluated. Tests on a soil artificially contaminated with copper chloride were carried out in order to evaluate the influence of the speed of percolation and of the chelating agent concentration (aqueous solution of an ethylendiaminotetraacetic acid di-sodium salt Na2-EDTA). At pH=7.3 an efficiency up to 93.9% for copper extraction was achieved by flushing 500 ml of Na2-EDTA 0.05 M solution and 100 ml of pure water at 0.792 cm/h. At these operating conditions the formation of EDTA complexes with other competitive cations (calcium and iron) was negligible. The experimental results were in agreement with the ones obtained using a model describing the chemistry of metal extraction. This model assessed that above pH=6 the formation of calcium and iron EDTA complexes was excluded and only the chelation of copper was allowed. The recovery of 91.6% of EDTA was also achieved by evaporating and acidifying the extracted solution: after filtration, solid EDTA was obtained, through the addition of sodium hydroxide Na2-EDTA. About 99.5% of the extracted copper was finally precipitated under alkaline conditions from the liquid phase. PMID- 16048736 TI - 'Biohyets': a method for displaying the extent and severity of environmental impacts. AB - Bioindicators are often more sensitive indicators of both biodiversity and environmental change than abiotic pollution parameters. The responses of selected plants and animals to anthropogenic insults can be used to assess environmental responses at a variety of spatial and temporal scales. This study maps the response of key reptile, mammal, bird and plant species to airborne contaminants around a large mine and mineral processing operation at Olympic Dam in arid Australia. The responses of different bioindicators should ideally be integrated in order to comprehend overall trends in biological integrity adjacent to pollution sources. Assimilation of different bioindicator responses allows greater precision and geographic coverage of the monitored region and reduces potential distortion from unrelated biological or monitoring responses of individual indicator groups. A single, integrated measure of ecosystem health that overlays the responses of otherwise incompatible datasets, is also of more value to industrial operators and environmental regulators than several disparate responses. Biohyets, which are the contours of bioindicator index values derived from multiple biotic measurements, are here used to map variability in ecosystem health and to identify regions, response variables and disturbance parameters for more rigorous analysis. PMID- 16048737 TI - Transaction costs and community-based natural resource management in Nepal. AB - Transaction costs in community-based resource management are incurred by households attempting to enforce property right rules over common resources similar to those inherent in private property rights. Despite their importance, transaction costs of community-based management of common pool resources (CPRs) are often not incorporated into the economic analysis of participatory resource management. This paper examines the transaction costs incurred by forest users in community forestry (CF) based on a survey of 309 households belonging to eight different forest user groups (FUGs) in the mid hills of Nepal. The analysis reveals that the average 'poor' household incurred Nepalese rupees (NRS) 1265 in transaction costs annually, while wealthier 'rich' households incurred an average of NRS 2312 per year. Although richer households bear higher proportions of such costs, transaction costs for CF management as a percentage of resource appropriation costs are higher for poorer households (26%) than those of middle wealth (24%) or rich households (14%). There are also village differences in the level of transaction costs. The results show that transaction costs are a major component of resource management costs and vary according to socio-economic status of resource users and characteristics of the community. PMID- 16048738 TI - Aged bovine chondrocytes display a diminished capacity to produce a collagen rich, mechanically functional cartilage extracellular matrix. AB - Most fundamental studies in cartilage tissue engineering investigate the ability of chondrocytes from young animals to produce cartilaginous matrix under various conditions, while current clinical applications such as autologous chondrocyte implantation, use chondrocytes from donors that are decades past skeletal maturity. Previous investigations have suggested that several characteristics of primary chondrocytes are age-dependent but none have quantified cell proliferation, proteoglycan synthesis and accumulation, collagen synthesis and accumulation, compressive and tensile mechanical properties in order to examine the effects of donor age on all of these parameters. We enzymatically isolated primary bovine chondrocytes from fetal, young and aged animals and cultured these cells in agarose gels to assess the above-mentioned properties. We found that fetal and young (but still skeletally mature i.e. 18-month-old bovine) chondrocytes behaved similarly, while aged chondrocytes (5- to 7-year-old bovine) displayed diminished proliferation ( approximately 2x less), a slightly reduced proteoglycan accumulation per cell ( approximately 20%), and significantly less collagen accumulation per cell ( approximately 55%) compared to the younger cells. Histological observations and mechanical properties supported these findings, where a particularly significant reduction in tensile stiffness produced by aged chondrocytes compared to younger cells was observed. Our findings suggest that donor age is an important factor in determining the outcome and potential success when tissue-engineered cartilage is produced from articular chondrocytes. More specifically, primary chondrocytes from aged donors may not possess sufficient capacity to produce the extracellular matrix that is required for a mechanically resilient tissue. PMID- 16048739 TI - Digestive enzyme patterns and evaluation of protease classes in Catla catla (family: Cyprinidae) during early developmental stages. AB - Digestive enzymes of Catla catla were studied during ontogenic development. Specific amylase activity was 0.12+/-0.01 mg maltose mg protein(-1) h(-1) in fish 4 days after hatching (DAH) and reached a maximum on (0.41+/-0.12 mg maltose mg protein(-1) h(-1)) 34 DAH. Total protease activity was minimum (123.2+/-16.5 mU mg protein(-1) min(-1)) on day-8 and reached its highest level (2713+/-147.2 mU mg protein(-1) min(-1)) on day-32. Trypsin activity showed constant increasing trend from day-16 onwards and was maximum on day-34 (118.1+/-7.09 mU mg protein( 1) min(-1)). Highest chymotrypsin activity was found on day-32 (1789.0+/-111.7 mU mg protein(-1) min(-1)). Lipase activity was detected in 4 DAH catla. Lipase activity increased steadily from day-22 onwards. SDS-PAGE of crude enzyme extracts showed that high molecular mass bands (41.8-127.8 kDa) appeared during the early stages followed by low molecular mass bands (17.8-37.2 kDa). The number of protease activity bands in substrate SDS-PAGE increased with age of fish. During ontogenesis of carp, soybean trypsin inhibitor (SBTI), PMSF and TLCK inhibited 75.5+/-1.19% to 92.8+/-0.85%, 53.3+/-9.47% to 90.5+/-2.6% and 39.8+/ 3.8% to 84.7+/-1.54% of total protease activity, respectively. There was only 2.58+/-0.66% to 10.21+/-0.09% inhibition of protease activity with EDTA. SBTI and PMSF inhibited 8 and 4 activity bands, respectively. TLCK, a specific trypsin inhibitor, inhibited four trypsin-like enzymes in carp during ontogenesis. PMID- 16048740 TI - Evaluation of postmortem urea nitrogen, creatinine and uric acid levels in pericardial fluid in forensic autopsy. AB - In postmortem biochemistry, there is insufficient data available for the practical analysis of factors in the pericardial fluid. The aim of the present study was to examine postmortem pericardial fluid for urea nitrogen (UN), creatinine (Cr) and uric acid (UA) levels to investigate the pathophysiology of death in forensic autopsy cases (total, n = 409; within 48 h postmortem), which included blunt, sharp instrument injury, asphyxiation, drowning, fire fatalities, hyperthermia, hypothermia, methamphetamine-related fatalities, other poisoning, delayed death from trauma and natural diseases. There was a significant elevation in the three markers for chronic renal failure, gastrointestinal bleeding, hyperthermia, hypothermia, methamphetamine fatalities and delayed traumatic death, which was comparable with the clinical criteria for their serum levels. These postmortem findings showed azotemia due to renal failure, elevated protein catabolism and rhabdomyolysis. Although the pericardial levels were otherwise similar to the clinical serum reference ranges, only the drowning fatalities showed significantly lower levels for each marker. These observations suggested the stability of UN, Cr and UA in the pericardial fluid within 48 h postmortem and their usefulness for the pathophysiological investigation of death involving azotemia. PMID- 16048741 TI - Plasmodium falciparum serine-repeat antigen (SERA) forms a homodimer through disulfide bond. AB - Plasmodium falciparum serine-repeat antigen (SERA) is one potential blood-stage vaccine candidate and is expressed as a protomer that is subsequently processed into four fragments (P47, P50, P6, and P17). Although recent evidence shows that P50 exhibits chymotrypsin-like protease activity, the function of SERA is still largely unknown. Here, we found that apart from cathepsin L-like cysteine protease, P50 showed significant homology to silicatein-alpha and testin which were shown to bind to cellular components, suggesting that SERA may have similar function. Immunoprecipitation of schizont lysate and molecular assignment of its precipitate by mass spectrometry provided evidence that SERA forms a homodimer through disulfide bond. Moreover, analysis of the fate of SERA using cell-free system revealed that the kinetics of conversion of SERA dimer into monomer is faster than that of processing of SERA monomer into various fragments. These findings may contribute to elucidate a possible function of SERA other than a protease. PMID- 16048742 TI - Predictors of exercise capacity and everyday activity in older heart failure patients. AB - BACKGROUND: Exercise capacity and daily activity are key outcomes for older, frail heart failure patients. Little is known about the determinants of these outcomes in this patient group. AIMS: To explore predictors of exercise capacity and daily activity in older, frail heart failure patients. METHODS: Analysis of prospectively collected data from a cohort of 82 patients aged 70 years and over, enrolled in a randomised controlled trial of exercise in heart failure patients. Pathophysiological, demographic, psychological and social factors were analysed by multivariate regression to determine predictors of exercise capacity (6-min walk distance) and daily activity (daily accelerometer counts). RESULTS: Between 49% and 55% of the variance in 6-min walk distance was explained by variables including New York Heart Association class, depression score, attitude to ageing and use of walking aids. Only 11% to 26% of the variance in accelerometer scores was explained by the model; 6-min walk distance was the only consistent predictor of daily activity. CONCLUSIONS: Physical, psychological and attitudinal variables contribute to variance of the 6-min walk. Six-minute walk distance predicts a small amount of the variance in daily activity, but the majority of variance in daily activity remains unexplained and requires further investigation. PMID- 16048743 TI - Natriuretic peptides in anthracycline-induced cardiotoxicity. PMID- 16048744 TI - Clinical trials update from the European Society of Cardiology Heart Failure meeting and the American College of Cardiology: darbepoetin alfa study, ECHOS, and ASCOT-BPLA. AB - This article provides information and a commentary on landmark trials presented at the European Society of Cardiology Heart Failure meeting held in June 2005, relevant to the pathophysiology, prevention and treatment of heart failure. All reports should be considered as preliminary data, as analyses may change in the final publication. The erythropoiesis stimulating protein, darbepoetin alfa, increased haemoglobin levels, improved quality of life and showed a trend for improved exercise duration in anaemic patients with symptomatic chronic heart failure. In the ECHOS study, the selective dopamine agonist nolomirole (CHF1035) showed no benefit in heart failure patients. Preliminary results of the ASCOT BPLA study, which were reported at the American College of Cardiology meeting in March 2005, showed that in hypertensive patients, treatment with a calcium antagonist plus an ACE inhibitor was more effective at reducing cardiovascular outcomes than atenolol plus a diuretic. PMID- 16048745 TI - Withholding or withdrawing intensive care treatment in paediatric bone marrow transplant: a moral maze. PMID- 16048746 TI - Consensus Conference Management of atopic dermatitis in children. Recommendations (short version). PMID- 16048747 TI - Anti-B- cell-directed immunotherapy (rituximab) in the treatment of refractory pemphigus--an update. AB - Rituximab is a monoclonal antibody directed against the CD20 surface antigen present on B lymphocytes. Following its application, B cells are rapidly and specifically depleted. Rituximab has been approved for the treatment of relapsed and therapy-refractory non-Hodgkin lymphoma and has been incorporated into numerous chemotherapy regimes with promising results. Eradication of auto reactive B cell clones is the rationale for its application in a variety of autoimmune disorders including the pemphigus group where B cells are thought to play a critical role in the pathogenesis. Preliminary reports in autoimmune disorders are encouraging. Adverse effects are generally well controlled and although severe infections have been reported following rituximab, the overall risk does not seem to be significantly increased. In the pemphigus group, rituximab has been successfully employed in refractory cases and a recent study suggests that a single course induces long-term remission in this patient group. PMID- 16048748 TI - Nevus roseus: a distinct vascular birthmark. AB - The new term nevus roseus is proposed to denote a lateralized telangiectatic birthmark with a light-red or pale-pink color, unlike the dark hue of nevus flammeus. It appears to be a distinct entity rather than just a color variant of nevus flammeus. Remarkably, nevus flammeus is a characteristic component of phacomatosis pigmentovascularis type II ("phacomatosis cesioflammea"), whereas nevus roseus represents a distinguishing feature of phacomatosis pigmentovascularis type III ("phacomatosis spilorosea"). In analogy to "port-wine stain" that is used as a synonym for nevus flammeus, nevus roseus could also be called "rose-wine stain". This lateralized vascular birthmark should be distinguished from the salmon patch that always involves the midline of the body. Nevus roseus belongs to the rather broad category of telangiectatic nevi, as well as to the even larger group of "capillary malformations". For obvious reasons these terms are not suitable to designate any specific type of vascular birthmark such as nevus roseus. PMID- 16048749 TI - Pathological score for the evaluation of allograft rejection in human hand (composite tissue) allotransplantation. AB - The experience obtained from the human hand allografts (HHA) performed to date suggests that the skin is a priviledged target of allograft rejection in this setting. The aim of this study was to establish a pathological score for assessing the severity of HHA rejection. The pathological slides of 89 skin biopsies obtained from the allografted limbs of six HHA recipients from day 0 to 5 years post-graft were retrospectively examined. According to the severity of the pathological changes observed, the following grades of rejection are proposed: 0: no rejection, I: mild rejection, II: moderate rejection, III: severe rejection, IV: very severe rejection. This grading system can be used as a basis for monitoring allograft rejection and for assessing the effects of the immunosuppressive treatment aiming at reversing HHA rejection; it can also be used for monitoring rejection of other skin-containing CTA. PMID- 16048750 TI - Changes of epidermal Langerhans cells in skin treated with trichloroacetic acid. AB - The changes in Langerhans cells (LCs) from normal human skin peeled with 40 and 60% trichloroacetic acid (TCA) or liquid nitrogen , which is known in cryosurgery as a control, were examined using monoclonal antibodies against the CD1a, HLA-DR and Lag in order to examine the immune surveillance system. In the 40% TCA group, the number of CD1a-positive cells decreased gradually until day 7, whereas both HLA-DR- and Lag-positive cells decreased for 12 hours, increased until day 2 and decreased thereafter. In the 60% TCA group, the number of CD1a-, HLA-DR- and Lag positive cells decreased gradually until day1, increased temporarily until day 2, and decreased again until day 7. There were no significant differences in the decrease of the LCs between the 40% and 60% TCA groups. In both cases the number of LCs on day 7 was statistically lower than before treatment. In the liquid nitrogen group, which served as a control, the LCs decreased gradually and slightly until day 2, and then increased. Taken together, the number of epidermal LCs from TCA-treated skin was reduced more significantly than LCs from liquid nitrogen-treated skin, suggesting a temporary impairment of the skin defense system. Therefore, long-term and frequent TCA peeling will require special attention for potential carcinogenesis. PMID- 16048751 TI - Comparative distribution of sulphur, thiols and disulphides in the porcine stratum corneum. AB - Biochemical, histochemical and cytochemical analyses were used to determine the sulphur contents and the thiol and disulphide distribution in the stratum corneum (SC) of the wild boar (WB), a large domestic pig breed (DP) and the Goettingen miniature pig (GMP). The sulphur contents (% DW) were different in the three animal types (WB: 1.70-1.38 body, 0.54 ear; DP: 0.84-0.53 body, 0.50 ear; GMP: 2.28-2.51 body, 2.66 ear). The results of the histochemical analysis of SH- and S-S- groups were clear, and densitometrical extinctions were highest in most body regions of the GMP for thiols and disulphides, followed by the DP for thiols, and the WB for disulphides. Absolute SC thickness was highest in the body of the GMP (62-80 mum), and generally lowest in the ear (20-38 mum) of all animal types. Relative SC thickness was the same for all animals in the body (40-66%), but lower in the ear (30%). Only -S-S- concentrations were correlated with SC thickness, and primarily in the GMP. Cytochemical analysis showed that high sulphur concentrations were obvious particularly in the CCE of corneal cells in the DP, as compared to the cytoplasm. Intracellular sulphur distribution was homogenous in the WB, and in the GMP, although in the latter at a higher concentration level. The results indicate breed-related effects on keratinisation in porcine corneal cells. Only the SC of the outer side of the ear of DP females is recommended as a model for humans. PMID- 16048752 TI - Gene expression profiling of peripheral blood mononuclear leukocytes from psoriasis patients identifies new immune regulatory molecules. AB - In the present report gene expression profiling of peripheral blood mononuclear cells (PBMC) from psoriasis patients suffering from severe generalized disease was performed comparing diseased stage with cured stage. By this means, 18 genes were identified which showed differential expression. The most significant differences were found for IL-8, annexin A3, cycloxygenase-2 (COX-2), cell cycle regulator G0S2, and pre-B cell enhancing factor (PBEF), all of which showed upregulation in the diseased stage. Microarray data were confirmed by real-time RT-PCR. Further analyses using support vector machines identified three pairs of genes (IL-8 - CDKN1C/p57, cyclooxygenase-2 - NR1D2, and desmocollin-2 - CDKN1C/p57) which allowed an accuracy of disease stage prediction of 86%, based on gene expression patterns. Taken together, this is the first large-scale gene expression study of psoriasis PBMC identifying candidate genes that might contribute to psoriasis immunopathogenesis. The genes identified in the present report and the molecular mechanisms underlying their regulation might serve as future targets for therapeutic intervention in psoriasis. PMID- 16048753 TI - Antifungal susceptibilities of dermatophytic agents isolated from clinical specimens. AB - The aim of this study was to investigate the susceptibility to four antifungal agents: ketoconazole, terbinafine, itraconazole and fluconazole, of the different species of dermatophyte strains isolated from clinical specimens. A total of 128 specimens were collected from toe nail, foot, inguinal region, trunk, hands and head. The dermatophytes tested included Trichophyton rubrum 108 (84.4%), Trichophyton mentagrophytes 11 (8.6%), Epidermophyton floccosum 5 (3.9%), Microsporum canis 2 (1.5%) and Trichophyton tonsurans 2 (1.5%). The mean minimum inhibitory concentrations (MIC) for the five species of dermatophytes ranged between 0.09-1.12 microg/mL for ketoconazole, 0.04-0.27 microg/mL for terbinafine, 0.08-0.43 microg/mL for itraconazole and 16.18-24.0 microg/mL for fluconazole. In vitro analysis of antifungal activity of these agents would also allow for the comparison between different systemic antifungals, which in turn may clarify the reasons for the lack of clinical response or serve as an effective therapy for patients with chronic infection. PMID- 16048754 TI - Syringotropic cutaneous T cell lymphoma treated with PUVA therapy. AB - Syringotropic cutaneous T cell Lymphoma (SCTCL) is a rare localized variant of CTCL. It is characterized by erythematous papules that, at histological examination, show dense dermal infiltrates of atypical T cells, that are preferentially located around hyperplastic eccrine sweat glands and ducts, with absent or minimal epidermotropism. Its relationship with mycosis fungoides and other CTCLs is not clarified and is still under discussion. Several treatment approaches have been suggested, but therapeutic results are often disappointing. We report the case of a patient with typical clinical and histopathological features of SCTCL and an excellent response to PUVA therapy. PMID- 16048755 TI - Lymphoma with large-plaque parapsoriasis treated with PUVA. AB - We report on a 78-year-old Japanese woman with a 50-year history of large-plaque parapsoriasis that had evolved into cutaneous T-cell lymphoma. Her large-plaque parapsoriasis had been treated with psoralen plus ultraviolet A for 10 years. Subsequently an isolated nodule appeared on her right lower leg. Prior or concurrent patches or plaques were absent. Histology revealed a diffuse nonepidermotropic infiltrate of large lymphocytes in the dermis, which had enlarged nuclei and prominent nucleoli. A diagnosis of CD30- cutaneous large T cell lymphoma was made. Following systemic chemotherapy, there was clinical improvement. No evidence of recurrence or systemic lymphoma has subsequently been found. PMID- 16048756 TI - Association of cutaneous side-effects of hydroxyurea and neuroendocrine carcinoma. AB - Many cutaneous side-effects of long term use of hydroxyurea (HU) therapy are well known. We report the first case, to our knowledge, of a neuroendocrine carcinoma located on the finger associated with other cutaneous changes due to this drug. Our patient received HU for two years for an thrombocytemia. This treatment was stopped because of a dermatomyositis-like eruption. Two years later, she developed multiple keratotic lesions, and eight years later, an Merkel cell carcinoma on the third left finger with metastasic evolution. This case suggests HU could be a cofactor of this neuroendocrine carcinoma but a simple coincidence could not be excluded. PMID- 16048757 TI - Two cases of chronic lymphoproliferative disorders in psoriatic patients treated with cyclosporine: hairy cell leukemia and Waldenstrom macroglobulinemia. AB - The real risk of lymphoproliferative disease in psoriatic patients has not yet been defined. Two explanations can be given for the occurrence of these malignancies: the broad immune activation typical of psoriasis and the administration of an immuno-suppressive treatment. A few studies describing the development of non Hodgkin lymphomas in psoriatic patients undergoing cyclosporine A have been published, but data about the occurrence of chronic lymphoproliferative disorders have never been reported. Here we describe the occurrence of hairy cell leukemia and Waldenstrom macroglobulinemia in two psoriatic patients treated with cyclosporine A. It remains unclear in our cases of chronic lymphoproliferative disease, as well as in the reported cases of psoriatic patients who develop lymphomas, whether psoriasis or the immunosuppressive treatment could play a role, although it is not possible to exclude a synergism between these factors. PMID- 16048758 TI - Results of a randomised, multicentre study comparing a new water-based gel of clindamycin 1% versus clindamycin 1% topical solution in the treatment of acne vulgaris. AB - Several topical formulations of clindamycin phosphate are currently marketed for the treatment of acne vulgaris. This 12 week, multi-centre, investigator-blind, randomised, active and placebo-controlled, parallel group study assessed the clinical efficacy and safety of clindamycin 1% gel once-a-day vs clindamycin 1% solution twice-a-day, and to demonstrate its superiority vs its vehicle alone. A total of 592 subjects were included. After 12 weeks, a 65% reduction in inflammatory lesion count was observed with both active treatments. The gel was superior to its vehicle for total and inflammatory lesion reduction, Global Assessment of Improvement, and Global Severity Grade at final visit (all p < 0.01). No difference was found between the 2 active treatments for any of the evaluated criteria. Local tolerance in each active treatment group was slightly better with clindamycin gel (1.9% of subjects) relative to 3.1% in the topical solution group. In conclusion, the new water-based gel once-a-day formulation of clindamycin 1% is an effective, safe, and convenient alternative to the twice-a day topical solution formulation in the treatment of acne vulgaris. PMID- 16048759 TI - Prevalence of psoriatic arthritis and joint complaints in a large population of Italian patients hospitalised for psoriasis. AB - We investigated the prevalence of psoriatic arthritis and joint complaints in a large population of Italian patients hospitalised for psoriasis. A total of 936 patients were evaluated for psoriatic arthritis according to the European Spondyloarthropathy Study Group (ESSG) criteria. They were visited by a dermatologist who evaluated specific joint signs and symptoms (i.e. arthralgia, stiffness, swelling, ankylosis, paresthesia) and assessed skin disease severity. Information on socio-demographic variables and other factors of clinical interest was also collected. Seventy-one patients (7.7% of the sample) had psoriatic arthritis (PsA). Overall, 90% of patients with PsA referred arthralgia, 70.4% stiffness, 67.7% swelling, 25.4% paresthesia, and 23.9% had ankylosis. However, among patients with plaque psoriasis who did not fulfil the criteria for PsA, 12.3% referred paresthesias, 7% arthralgia, 4.2% stiffness, 3.7% swelling, and 1.2% had ankylosis. Among 114 patients referring arthralgia, 42 (39.6%) did not meet the ESSG criteria for PsA. Therefore, patients with plaque psoriasis refer joint symptoms quite frequently making it difficult to distinguish patients with psoriatic arthritis from patients with joint complaints. PMID- 16048760 TI - Make-up improves the quality of life of acne patients without aggravating acne eruptions during treatments. AB - Boehncke et al. suggested that decorative cosmetics can improve the quality of life (QOL) of skin diseases. But dermatologists sometimes discourage female acne patients from applying make-up since decorative cosmetics are considered one of the aggravating factors for acne eruptions. The purpose of this study is to assess whether make-up application interferes with acne treatments and how QOL changes when the make-up items are designed for acne patients and used in order to disguise acne eruptions. Eighteen female acne patients were trained by a make up artist and advised to apply acne-designed basic and decorative cosmetics for 2 to 4 weeks while their acne was appropriately treated. The acne-severity and QOL of patients were assessed before and after the study. The results revealed that the number of acne eruptions decreased even though patients were applying make up. The QOL scores of Skindex-16, GHQ30 and anxiety state index greatly improved. Our results suggest that dermatologists should encourage acne patients to utilize appropriate make-up to improve their QOL. PMID- 16048761 TI - Transient intra-epidermal bullous dermatosis. AB - Transient acantholytic dermatosis is an acquired, mildly pruritic, papulovesicular disease affecting the trunk and extremities. In this report a case of the bullous variant of this disease is described, with a brief review of the literature. PMID- 16048762 TI - Eccrine angiomatous hamartoma with massive mucin deposition. AB - Eccrine angiomatous hamartoma (EAH) is a rare condition characterized histologically by an increased number of eccrine structures and numerous vascular elements. Although EAH sometimes shows mucinous stroma, the mucin is usually not abundant. In this report, we described a case of EAH that had massive mucin deposition, especially around the eccrine structures. Our case and a similar reported one indicate that EAH includes a rare variant with a large amount of mucin as one of the hamartoma components. PMID- 16048763 TI - Fatal herpes simplex virus infection in Darier disease under corticotherapy. AB - A 67-year-old man is presented with longstanding and severe Darier disease treated by topical antiseptics and potent corticosteroids, in combination with oral glucocorticoids and etretinate. After cardiac bypass surgery in 1997, the patient experienced herpes simplex virus (HSV type-1) infection of the skin that was treated by intravenous aciclovir. In 2003, he presented a widespread atypical exacerbation of his Darier disease, involving the face, trunk, buttocks, intertriginous areas and arms. Initial clinical signs and bacteriological findings suggested a bacterial involvement by multiresistant Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Proteus mirabilis. Despite antibiotherapy, the clinical presentation progressively worsened. A skin biopsy was performed and immunohistochemical examination identified a type-2 HSV infection. Although intravenous aciclovir was administered, the widespread cutaneous HSV infection was followed by systemic dissemination. A severe acute respiratory distress syndrome (ARDS) developed, leading to a fatal issue. At autopsy, a severe interstitial type-2 HSV pneumonitis with extensive necrotic areas was found, in association with gastro-intestinal involvement. This case represents, to the best of our knowledge, the first case of Darier disease presenting a fatal type-2 HSV infection. It underlines the importance of rapidly recognizing HSV infection in Darier disease and stresses the risk of lethal outcome. The different risk factors for HSV infection in this patient are reviewed. PMID- 16048764 TI - Disseminated superficial porokeratosis with amyloid deposits. AB - We report a 65-year-old Caucasian man who exhibited a disseminated superficial porokeratosis. Histological examination with haematoxylin and eosin stain revealed the presence of dermal deposits of an eosinophilic amorphous material that stained positively with crystal violet and showed bright yellow fluorescence with thioflavin-T under the polarizing microscopy, indicating that it was amyloid. Colloid bodies from porokeratotic lesions can degenerate into amyloid, but it is not usually detected with haematoxylin and eosin stains because of its small quantity. Probably amyloid deposits could be detected more frequently in porokeratosis if histochemical techniques were carried out in a routine way. PMID- 16048765 TI - Granular-cell basal cell carcinoma of the skin. AB - Granular cell basal cell carcinoma (GBCC) is a very rare variant of BCC, of which ten cases have been reported in the literature. We describe here a new case of GBCC studied immunohistochemically. The tumor developed on the face of a 71-year old man and showed typical features of GBCC, i.e. a tumor reminiscent of nodular BCC consisting of cells with a granular eosinophilic cytoplasm. Immunohistochemically, tumor cells expressed cytoplasmic reactivity for keratins, CD68 and CD15 antigens, bcl-2 oncoprotein and nuclear reactivity for the p63 protein. Based on a review of the literature, the salient features of GBCC are discussed. PMID- 16048766 TI - Problems with mitochondrial DNA as a marker in population, phylogeographic and phylogenetic studies: the effects of inherited symbionts. AB - Mitochondrial DNA (mtDNA) has been a marker of choice for reconstructing historical patterns of population demography, admixture, biogeography and speciation. However, it has recently been suggested that the pervasive nature of direct and indirect selection on this molecule renders any conclusion derived from it ambiguous. We review here the evidence for indirect selection on mtDNA in arthropods arising from linkage disequilibrium with maternally inherited symbionts. We note first that these symbionts are very common in arthropods and then review studies that reveal the extent to which they shape mtDNA evolution. mtDNA diversity patterns are compatible with neutral expectations for an uninfected population in only 2 of 19 cases. The remaining 17 studies revealed cases of symbiont-driven reduction in mtDNA diversity, symbiont-driven increases in diversity, symbiont-driven changes in mtDNA variation over space and symbiont associated paraphyly of mtDNA. We therefore conclude that these elements often confound the inference of an organism's evolutionary history from mtDNA data and that mtDNA on its own is an unsuitable marker for the study of recent historical events in arthropods. We also discuss the impact of these studies on the current programme of taxonomy based on DNA bar-coding. PMID- 16048767 TI - Nocturnal life of young songbirds well before migration. AB - In songbirds, nocturnal activity is believed to be a characteristic feature of migration. However, unlike experimental conditions where the onset of nocturnal restlessness is defined as a shift of activity leading up to the dark period, this behaviour has, until now, not been observed in natural conditions. Here we studied the nocturnal behaviour of radio-tagged juvenile Eurasian reed warblers (Acrocephalus scirpaceus) during the pre-migratory period. The birds started nocturnal flights at the age of 38 days, whereas migration did not commence until they were at least 50 days old. The birds left their natal site by nocturnal flights and repeatedly returned to it. Such shuttle movements suggest the existence of a previously unknown period of nocturnal activity. Motivation to perform such night flights gradually increases with age. We relate the function of these nocturnal pre-migratory flights to the development of a stellar compass, necessary for detecting the compass direction towards winter quarters and for the formation of a navigational target, which will be used during return (spring) migration. PMID- 16048768 TI - Female Bicyclus anynana butterflies choose males on the basis of their dorsal UV reflective eyespot pupils. AB - Sexual and natural selection pressures are thought to shape the characteristic wing patterns of butterfly species. Here we test whether sexual selection by female choice plays a role in the maintenance of the male wing pattern in the butterfly Bicyclus anynana. We perform one of the most extensive series of wing pattern manipulations in butterflies, dissecting every component of the 'bulls eye' eyespot patterns in both ventral and dorsal wing surfaces of males to test the trait's appeal to females. We conclude that females select males on the basis of the size and brightness of the dorsal eyespot's ultraviolet reflecting pupils. Pupil absence is strongly selected against, as are artificially enlarged pupils. Small to intermediate (normal sized) pupils seem to function equally well. This work contradicts earlier experiments that suggest that the size of dorsal eyespots plays a role in female choice and explains why male dorsal eyespots are very variable in size and often have indistinct rings of coloration, as the only feature under selection by females seems to be the central white pupil. We propose that sexual selection by female choice, rather than predator avoidance, may have been an important selective factor in the early stages of eyespot evolution in ancestral Lepidopteran lineages. PMID- 16048769 TI - Behaviour of leatherback sea turtles, Dermochelys coriacea, during the migratory cycle. AB - Leatherback sea turtles, Dermochelys coriacea, undertake broad oceanic movements. While satellite telemetry has been used to investigate the post-nesting behaviour of female turtles tagged on tropical nesting beaches, long-term behavioural patterns of turtles of different sexes and sizes have not been described. Here we investigate behaviour for 25 subadult and adult male and female turtles satellite tagged in temperate waters off Nova Scotia, Canada. Although sex and reproductive condition contributed to variation in migratory patterns, the migratory cycle of all turtles included movement between temperate and tropical waters. Marked changes in rates of travel, and diving and surfacing behaviour, accompanied southward movement away from northern foraging areas. As turtles approached higher latitudes the following spring and summer, they assumed behaviours consistent with regular foraging activity and eventually settled in coastal areas off Canada and the northeastern USA. Behavioural patterns corresponding to various phases of the migratory cycle were consistent across multiple animals and were repeated within individuals that completed return movements to northern waters. We consider the potential biological significance of these patterns, including how turtle behaviour relates to predator avoidance, thermoregulation and prey distribution. PMID- 16048770 TI - The role of optimal vortex formation in biological fluid transport. AB - Animal phyla that require macro-scale fluid transport for functioning have repeatedly and often independently converged on the use of jet flows. During flow initiation these jets form fluid vortex rings, which facilitate mass transfer by stationary pumps (e.g. cardiac chambers) and momentum transfer by mobile systems (e.g. jet-propelled swimmers). Previous research has shown that vortex rings generated in the laboratory can be optimized for efficiency or thrust, based on the jet length-to-diameter ratio (L/D), with peak performance occurring at 3.5